CN101934036B - Method for preparing medicament for treating diabetes mellitus - Google Patents
Method for preparing medicament for treating diabetes mellitus Download PDFInfo
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Abstract
The invention discloses a method for preparing a medicament for treating diabetes mellitus, which comprises the following steps of: extracting six medicinal materials, namely southern schisandra, kudzuvine root and the like respectively, performing macroporous resin purification on the six medicinal materials of the kudzuvine root and the like, concentrating extracting solution, mixing, drying, pelletizing, and adding glibenclamide to prepare an oral administration preparation. Active ingredients of traditional Chinese medicines are extracted furthest by the new preparation method, and invalid ingredients are removed to fulfill the aims of reducing dosage along with convenient administration and carrying; simultaneously, the method ensures that a process meets the requirement of large-scale production.
Description
Technical field
The present invention relates to a kind of preparation technology of Remedies for diabetes.
Background technology
Diabetes, namely alleged " diabetes " of the traditional Chinese medical science is a kind of common incretion metabolism disease take hyperglycemia as common trait, absolute or relative deficiency causes owing to insulin.The name of disease of quenching one's thirst, head sees " the strange sick opinion of Plain Questions ", and " Medical Treasures of the Golden Chamber " vertical specially piece of writing is discussed, and proposes treatment side's medicine the earliest." Standards of Diagnosis and Treatment disappear Shan " have carried out standard to the clinical classification of three types of diabetes on the basis that forefathers discuss, " thirsty and polydipsia disappear on being (through the meaning diabetes involving the upper-JIAO); Rapid digestion of food and polyorexia disappear in being (in meaning disappears); Yearningly and just number cream is arranged is lower disappearing (through calling diabetes involving the kidney) ".Bright clear after, Therapeutic Principle and square medicine to quenching one's thirst have had research more extensively and profoundly.To the understanding of Pathogenesis of Diabetes, the most representative at present is the deficiency of both QI and YIN theory: think that the primary disease pathogeny is scorching impairment of YIN, the cloudy QI consumed of decreasing causes deficiency of both QI and YIN, when take supplementing QI and nourishing YIN as method for the treatment of.
Diabetes pill is that the present patent application people forms at ancient prescription " Yuquan is loose " and the basis sanction of " the side of quenching one's thirst ", full side is comprised of Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis, Fructus Schisandrae Sphenantherae, Rhizoma Dioscoreae, glibenclamide etc., has nourishing kidney-YIN, the effect of supplementing QI for promoting the production of body fluid, be used for type of deficiency of both QI and YIN diabetes (type 2 diabetes mellitus), disease is seen thirst and liking drink, polyuria, polyorexia, is become thin, the patient of fatigue and asthenia, the lazy speech of breathing hard.The present patent application people's Chinese invention patent ZL200610075069.6 discloses a kind of prescription for the treatment of diabetes and has formed and preparation technology.In the pharmaceutical composition of this patent, its preparation method is part Chinese crude drug water extraction in the prescription, make various dosage forms behind the extract obtained fine powder mix homogeneously that other medical materials are worn in prescription, technique is tradition comparatively, extracts active ingredients is (especially Fructus Schisandrae Sphenantherae) not exclusively, and also contain some invalid components, thereby caused the larger shortcoming of dosage, make troubles for patient's long-term taking.
Summary of the invention
Large in order to solve dose, be unsuitable for the problem of long-term taking, the invention provides a kind of process for preparing medicine for the treatment of diabetes.
For achieving the above object, the present invention has taked following technical scheme:
A kind of preparation method of Remedies for diabetes, the active component of this pharmaceutical composition is to be made by the crude drug of following ratio: Radix Rehmanniae 600-1200 weight portion, Radix Astragali 200-400 weight portion, Rhizoma Dioscoreae 100-200 weight portion, Radix Puerariae 1000-2000 weight portion, Radix Trichosanthis 1000-2000 weight portion, Stigma Maydis 1000-2000 weight portion, Fructus Schisandrae Sphenantherae 200-400 weight portion, glibenclamide 0.2-1.5 weight portion may further comprise the steps:
(1) get the Fructus Schisandrae Sphenantherae medicinal material coarse powder, extract 1～3 time with 60～95% alcohol heating reflux of 4～8 times of crude drug amounts, each 1～3 hour, crossing the concentrated relative density that obtains of leaching filtrate decompression was 1.05～1.2 concentrated solution 1;
(2) get Radix Rehmanniae, the Radix Astragali, Rhizoma Dioscoreae, Radix Puerariae, Radix Trichosanthis, Stigma Maydis, extract 1～3 time with 40～95% alcohol heating reflux of 6～12 times of crude drug amounts, each 1～2 hour, it was 1.05～1.2 concentrated solution 2 that the extracting solution concentrating under reduced pressure obtains relative density;
(3) concentrated solution 2 is injected in the macroporous adsorptive resins of having handled well slowly, first water is that eluent is washed till the Molish reaction and is negative, and discards eluent; Use again 20～80% ethanol elutions of 2～4 times of cylinder accumulated amounts, collect the 20-80% ethanol elution, Recycled ethanol and the concentrated concentrated solution 3 that obtains relative density 1.05～1.2;
(4) with concentrated solution 3 and concentrated solution 1 mix homogeneously, vacuum drying is pulverized granulation;
(5) add glibenclamide and various conventional adjuvant, such as disintegrating agent, lubricant, binding agent etc., method of Chinese medicinal prepares and get final product routinely.
Described medicine is oral formulations, such as tablet, capsule or pill.
Preferably, described step (1) is extracted 2 times for 80% alcohol heating reflux with 6 times of crude drug amounts, and each 2 hours, the relative density of described concentrated solution 1 was 1.10～1.15.
Preferably, described step (2) is extracted 3 times for 50% alcohol heating reflux with 8 times of crude drug amounts, and each 1 hour, the relative density of described concentrated solution 2 was 1.05～1.10.
Preferably, the technological parameter of macroporous adsorbent resin is in the described step (3): medical material: resin is 1: 1; Absorption flow velocity and elution flow rate be 3 times of column volumes/hour; The blade diameter length ratio of post bed is 1: 3; The concentration of ethanol is 40%, and consumption is 3 times of cylinder accumulated amounts.
The active ingredient of medicine of the present invention is made by crude drug such as Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis, Fructus Schisandrae Sphenantherae, Rhizoma Dioscoreae, glibenclamide.With the Radix Rehmanniae nourishing kidney-YIN, clearing away heat and promoting production of body fluid is monarch in the side, take Radix Puerariae, the Radix Astragali, Radix Trichosanthis, Fructus Schisandrae Sphenantherae, Rhizoma Dioscoreae supplementing QI and nourishing YIN, promoting the production of body fluid to quench thirst as minister; Assistant expels the heat-evil with the Stigma Maydis diuresis; All medicines share, and play altogether nourishing kidney-YIN, the effect of supplementing QI for promoting the production of body fluid.Clinical in the type of deficiency of both QI and YIN type 2 diabetes mellitus.Medicine of the present invention is on the basis of original pharmaceutical composition, thereby takes advanced separation purifying technique to reduce dosing, has improved product quality.
Medicine of the present invention is oral formulations, and is oral, and take tablet as example, a 1-2 sheet 2-3 time on the one, is used warm water delivery service ante cibum.Convert every and contain crude drug amount 5.43g, the crude drug amount that contains with per 5 diabetes pilles is suitable.Onset dosage is 3g crude drug/kg; Effective dosage ranges is 3g crude drug/kg～12g crude drug/kg; Onset time is 4～6 weeks of administration; The best use of time is 6 weeks of administration; Duration of efficacy is 4～8 weeks of administration.
Compared with prior art, the present invention has following remarkable result:
1. the present invention extracts the prescription medical material respectively by new preparation method, and macroporous resin adsorption technique is carried out purification, and the active ingredient with Chinese medicine extracts to greatest extent, removes invalid element, thereby reaches the raising curative effect, reduces the purpose of dose; Guaranteed that simultaneously technique meets the needs of large production;
2. the dose of medicine of the present invention obviously reduces, but effect and prior art are suitable, take tablet as example, once only need to take 1～2, and existing pharmaceutical composition (diabetes pill) need to be taken 5～10, and therefore, drug administration of the present invention makes things convenient for, is easy to carry;
3. drug quality of the present invention is controlled, stable, and pharmacodynamic result is suitable with former pharmaceutical composition, and toxicologic study is not found toxic and side effects.
Description of drawings
Fig. 1 is the elution curve of HPD400A;
Fig. 2 is the elution curve of HPD100;
Fig. 3 is the amount of puerarin in each part of 40%EtOH eluent.
The specific embodiment
Describe the present invention in detail below in conjunction with specific embodiment.
The preparation technology of embodiment 1 medicinal tablet of the present invention
The active component of present embodiment Chinese medicine compositions is made (1000) by the crude drug of following ratio
Radix Puerariae 1325g Radix Rehmanniae 795g Radix Astragali 265g Radix Trichosanthis 1325g
Stigma Maydis 1325g Fructus Schisandrae Sphenantherae 265g Rhizoma Dioscoreae 132.5g glibenclamide 1.25g
The preparation method of medicinal tablet of the present invention is: may further comprise the steps:
(1) get the Fructus Schisandrae Sphenantherae medicinal material coarse powder of recipe quantity, 80% alcohol heating reflux that adds 6 times of crude drug amounts extracts 2 times, each 2 hours, filters.Merge 2 times filtrate, decompression recycling ethanol also is concentrated into the concentrated solution 1 that relative density is 1.10～1.15 (50 ℃), and is for subsequent use;
(2) get other 6 flavor medical material (Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis, Rhizoma Dioscoreae) of recipe quantity, 50% alcohol heating reflux that adds 8 times of crude drug amounts extracts 3 times, each 1 hour, filters.Merge 3 times filtrate, decompression recycling ethanol also is concentrated into the concentrated solution 2 that relative density is 1.05～1.10 (50 ℃);
(3) with (main technologic parameters of purification with macroreticular resin: medical material: resin is 1: 1 in the concentrated solution 2 HPD100 macroporous adsorptive resins that injection has been handled well slowly; Absorption flow velocity and elution flow rate be 3 times of column volumes/hour; The blade diameter length ratio of post bed is 1: 3), wash with water first to Molish reaction and be negative, discard water elution liquid; Use 40% ethanol elution of 3 times of cylinder accumulated amounts again, collect 40% ethanol elution, Recycled ethanol also is concentrated into relative density 1.10～1.15 (50 ℃);
(4) add above-mentioned Fructus Schisandrae Sphenantherae concentrated solution 1, mix homogeneously, vacuum drying (80 ℃) is pulverized (crossing 80 mesh sieves), obtains extract powder; Extract powder is put in the fluidised bed granulator, sprayed into PVP K30 solution and granulate, the control pellet moisture is sifted out 16～60 purpose granules about 5～7%, weigh;
(5) will make granule puts in the fluidised bed granulator; take by weighing glibenclamide (Tianjin Inst. of Materia Medica pharmaceutcal corporation, Ltd) according to grain amount by proportioning in the prescription; add PVP K30 (Huzhou Zhanwang Pharmaceutical Co., Ltd.) solution and make suspension; be sprayed on the granule; dried particles to moisture about 5～7%, take out.Add polyvinylpolypyrrolidone (American I SP company), carboxymethylstach sodium (Huzhou Zhanwang Pharmaceutical Co., Ltd.), silicon dioxide (Rhizoma Dioscoreae industry company limited in the Zhejiang), magnesium stearate (the honorable chemical industry company limited in Tongzi county, Guizhou Province), mixing, tabletting;
(6) take by weighing in proportion required film coating powder (85G61081, Shanghai Colorcon Coating Technology Co., Ltd), in liquid dispensing container, add required purified water, place the central authorities of container to be positioned at 2/3 place under the liquid level stirring paddle.Start stirring paddle, liquid can be stirred adjusting rotary speed fully and liquid level just forms whirlpool.The film coating powder is spread on the liquid level with speed stably, and speed can be involved in rapidly whirlpool with powder is advisable, and improves in case of necessity mixing speed to keep stable vorticity.After all film coating powder all add, reduce mixing speed, whirlpool is disappeared.Continue to stir 45 minutes, get final product.Label is put into coating pan be heated to about 40 ℃, spray into coating solution in the rotation, until it is complete to whitewash, dry getting final product.
The preparation technology of embodiment 2 medicine capsules of the present invention
The active component of present embodiment Chinese medicine compositions is made (1000) by the crude drug of following ratio
Radix Puerariae 1325g Radix Rehmanniae 795g Radix Astragali 265g Radix Trichosanthis 1325g
Stigma Maydis 1325g Fructus Schisandrae Sphenantherae 265g Rhizoma Dioscoreae 132.5g glibenclamide 1.25g
The preparation method of medicine capsule of the present invention is: may further comprise the steps:
(1) get the Fructus Schisandrae Sphenantherae medicinal material coarse powder of recipe quantity, 60% alcohol heating reflux that adds 4 times of crude drug amounts extracts 1 time, each 3 hours, filters.Decompression recycling ethanol also is concentrated into the concentrated solution 1 that relative density is 1.05～1.15 (50 ℃), and is for subsequent use;
(2) get other 6 flavor medical material (Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis, Rhizoma Dioscoreae) of recipe quantity, 40% alcohol heating reflux that adds 6 times of crude drug amounts extracts 1 time, each 2 hours, filters.Decompression recycling ethanol also is concentrated into the concentrated solution 2 that relative density is 1.05～1.10 (50 ℃);
(3) with (main technologic parameters of purification with macroreticular resin: medical material: resin is 1: 1 in the concentrated solution 2 HPD100 macroporous adsorptive resins that injection has been handled well slowly; Absorption flow velocity and elution flow rate be 3 times of column volumes/hour; The blade diameter length ratio of post bed is 1: 3), wash with water first to Molish reaction and be negative, discard water elution liquid; Use 80% ethanol elution of 2 times of cylinder accumulated amounts again, collect 80% ethanol elution, Recycled ethanol also is concentrated into relative density 1.10～1.2 (50 ℃);
(4) add above-mentioned Fructus Schisandrae Sphenantherae concentrated solution 1, mix homogeneously, vacuum drying (80 ℃) is pulverized (crossing 80 mesh sieves), obtains extract powder; Extract powder is put in the fluidised bed granulator, sprayed into PVP K30 solution and granulate, the control pellet moisture is sifted out 16～60 purpose granules about 5～7%, weigh;
(5) will make granule puts in the fluidised bed granulator; take by weighing glibenclamide (Tianjin Inst. of Materia Medica pharmaceutcal corporation, Ltd) according to grain amount by proportioning in the prescription; add PVP K30 (Huzhou Zhanwang Pharmaceutical Co., Ltd.) solution and make suspension; be sprayed on the granule; dried particles to moisture about 5～7%; take out, capsule is made in filling.
The preparation technology of embodiment 3 bolus of drug of the present invention
The active component of present embodiment Chinese medicine compositions is made (1000 ball) by the crude drug of following ratio
Radix Puerariae 1325g Radix Rehmanniae 795g Radix Astragali 265g Radix Trichosanthis 1325g
Stigma Maydis 1325g Fructus Schisandrae Sphenantherae 265g Rhizoma Dioscoreae 132.5g glibenclamide 1.25g
The preparation method of bolus of drug of the present invention is: may further comprise the steps:
(1) get the Fructus Schisandrae Sphenantherae medicinal material coarse powder of recipe quantity, 85% alcohol heating reflux that adds 8 times of crude drug amounts extracts 3 times, each 1 hour, filters.Merge 3 times filtrate, decompression recycling ethanol also is concentrated into the concentrated solution 1 that relative density is 1.10～1.2 (50 ℃), and is for subsequent use;
(2) get other 6 flavor medical material (Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis, Rhizoma Dioscoreae) of recipe quantity, 95% alcohol heating reflux that adds 12 times of crude drug amounts extracts 2 times, each 1 hour, filters.Merge 2 times filtrate, decompression recycling ethanol also is concentrated into the concentrated solution 2 that relative density is 1.10～1.2 (50 ℃);
(3) with (main technologic parameters of purification with macroreticular resin: medical material: resin is 1: 1 in the concentrated solution 2 HPD100 macroporous adsorptive resins that injection has been handled well slowly; Absorption flow velocity and elution flow rate be 3 times of column volumes/hour; The blade diameter length ratio of post bed is 1: 3), wash with water first to Molish reaction and be negative, discard water elution liquid; Use 20% ethanol elution of 4 times of cylinder accumulated amounts again, collect 20% ethanol elution, Recycled ethanol also is concentrated into relative density 1.05～1.15 (50 ℃);
(4) add above-mentioned Fructus Schisandrae Sphenantherae concentrated solution 1, mix homogeneously, vacuum drying (80 ℃) is pulverized (crossing 80 mesh sieves), obtains extract powder;
(5) use the one-step-granulating method granulation, add glibenclamide and an amount of PVP K30, be pressed into pill.
Medicinal tablet preparation technology of the present invention extracts first purification with the Chinese medicine in the prescription, through boiling granulating and then spray into suspendible glibenclamide, tabletting.Because the main effective ingredient of the Fructus Schisandrae Sphenantherae in the prescription is Lignanoids compounds, polarity is less, fat-soluble larger, add that Fructus Schisandrae Sphenantherae medical material proportion in whole prescription is smaller, in order better to bring into play the curative effect of Fructus Schisandrae Sphenantherae, the present invention is its independent extraction, and then will merge with the extract of other medical material.
A, Fructus Schisandrae Sphenantherae extract research
Schisantherin A is one of main effective ingredient of Fructus Schisandrae Sphenantherae.Lignan component has stronger fat-soluble, can be dissolved in the organic solvents such as methanol, ethanol, acetone, ethyl acetate.In extracting solvent, selected water, 50% ethanol, 80% ethanol to carry out the analysis on the impact of schisantherin A extraction ratio.
Take by weighing Fructus Schisandrae Sphenantherae medical material and each 30g of Fructus Schisandrae Sphenantherae medicinal material coarse powder of not pulverizing, 80% alcohol heating reflux that adds respectively 6 times of crude drug amounts extracted 1 hour, filtered, filtrate decompression is concentrated, and standardize solution shakes up in the 100ml measuring bottle respectively, carries out HPLC and detect after 0.45 μ m microporous filter membrane filters.2 parts of Duplicate Samples the results are shown in Table 1.
The different degree of grinding of table 1 medical material are on the impact of schisantherin A extraction ratio
The result shows: the extraction ratio whether medical material is pulverized schisantherin A has a great impact, and the extraction ratio of coarse powder is higher than not grinding medicinal materials far away, so medical material will be ground into coarse powder.
Take by weighing 3 parts of Fructus Schisandrae Sphenantherae medicinal material coarse powder, every part of 30g adds respectively entry, 50% ethanol, each 100ml of 80% ethanol, supersound extraction 20 minutes filters, and filtrate decompression is concentrated, standardize solution shakes up in the 50ml measuring bottle respectively, carries out HPLC and detect after 0.45 μ m microporous filter membrane filters.2 parts of Duplicate Samples the results are shown in Table 2.
Table 2 different solvents is on the impact of schisantherin A extraction ratio
--: do not detect schisantherin A.
The result shows: the extraction effect of ethanol is better than water extraction, and the concentration of ethanol is higher, and extraction effect better.Therefore, tentatively first with 80% ethanol Fructus Schisandrae Sphenantherae is extracted.
Investigated three kinds of extracting method commonly used: warm macerating method, ultrasonic extraction and heating and refluxing extraction method, method is as follows:
The warm macerating method: the material coarse powder 30g that gets it filled,, filter 50 ℃ of lower immersions 1 hour with 80% ethanol of 6 times of medical material amounts, filtrate decompression is concentrated, and is settled in the 100ml measuring bottle, and is for subsequent use.
Ultrasonic extraction: the material coarse powder 30g that gets it filled, with 6 times of medical material amounts, 80% ethanol ultrasonic extraction 1 hour, filter, filtrate decompression is concentrated, and is settled in the 100ml measuring bottle, and is for subsequent use.
Reflux extraction: the material coarse powder 30g that gets it filled, extracted 1 hour with 6 times of medical material amount 80% alcohol heat reflux, filter, filtrate decompression is concentrated, and is settled in the 100ml measuring bottle, and is for subsequent use.
Each extracting solution carries out HPLC and detects after 0.45 μ m microporous filter membrane filters.2 parts of Duplicate Samples the results are shown in Table 3.
The investigation result of table 3 Different Extraction Method
Above result shows: the amount of the schisantherin A that heating reflux method extracts is maximum, and suitable commercial production, so extracting method is selected heating reflux method.
Take ethanol as extracting solvent, the major influence factors that adopts heating reflux method to extract lignan component in the Fructus Schisandrae Sphenantherae has: concentration of alcohol (A), extraction time (B), extraction time (C), ethanol consumption (D), still selected the orthogonal test table of four factors, three levels to test, and take must the measuring as evaluation index of schisantherin A, preferred optimum extraction process.Factor, water-glass see Table 4, and orthogonal test table sees Table 5.
Table 4 factor level table
Take by weighing Fructus Schisandrae Sphenantherae medicinal material coarse powder 50g, test by each test item that table 5 is listed, merge extractive liquid,, concentrating under reduced pressure, vacuum drying obtains extract.Precision takes by weighing this extract 50mg, places the 25ml measuring bottle, adds methanol and is diluted to scale, shakes up, and carries out HPLC and detect after 0.45 μ m microporous filter membrane filters.Adopt one point external standard method, calculate must measuring of schisantherin A in the 50g medical material, the results are shown in Table 5.
Table 5 orthogonal test table and result
From intuitive analysis (seeing Table 5): factor A (concentration of alcohol) and factor D (ethanol consumption) are larger on the stripping impact of schisantherin A, secondly are factor B (extraction times), and the impact of factor C (extraction time) is minimum.During concentration of alcohol high (95%), extract not exclusively, although the percentage composition of schisantherin A is higher, but the extractum amount that extracts is few, productive rate is low, and when extracting with low concentration alcohol (60%), the impurity that some polarity are larger extracts easily, cause the content of schisantherin A on the low side, therefore can select A
2, namely 80% ethanol extraction is for well; The extraction effect that adds 8 times of ethanol and 6 times of ethanol is better than the extraction effect of 4 times of ethanol, considers industrial cost, can select D
2Extraction time is with B
2For good, the time, then extracted not exclusively short (1 hour), and overlong time (3 hours) then may make schisantherin A change, and extraction time is selected C
3
Table 6 analysis of variance table
From variance analysis (seeing Table 6), difference that there are no significant between each level of each factor.Therefore consider extraction time selection C from the cost angle
2, in conjunction with the result of intuitive analysis and variance analysis, extraction conditions is selected A
2B
2C
2D
2, namely 80% alcohol heating reflux with 6 times of crude drug amounts extracts each 2 hours 2 times.
According to above optimal extract process, carry out 2 batches of process certification tests, the results are shown in Table 7.
Table 7 extraction process demonstration test result
The result show this process stabilizing, reliable, repeatability is good.Therefore, determine the extraction process of Fructus Schisandrae Chinensis to be: 80% alcohol heating reflux with 6 times of crude drug amounts extracts 2 times, other 6 medicinal material extract research of distinguishing the flavor of such as each 2 hours B, Radix Puerariae
The blood sugar lowering effective ingredient of Radix Puerariae mainly is puerarin, but but also alcohol extraction of water extraction; Studies show that behind the water extract, ethanol extract, water extraction of Radix Rehmanniae and behind the 60% ethanol precipitate with ethanol blood sugar reducing function is arranged all; There is research to think that Stigma Maydis hypoglycemic activity composition is mainly saponin; The main effective ingredient of the Radix Astragali is saponin and polysaccharide, and the ratio of polysaccharide component is also very large in Rhizoma Dioscoreae and the Radix Trichosanthis, therefore in order to improve the extraction ratio of effective ingredient, reduces paste-forming rate, adopts 50% ethanol extraction.Consider that Radix Puerariae proportion in whole prescription is larger, and according to the literature, puerarin is one of main effective ingredient of diabetes pill, and therefore we mainly adopt the high performance liquid chromatogram method to measure the content of puerarin in the process study process, as one of index of estimating extraction effect.Simultaneously, the astragaloside in the employing TLC method detection Milkvetch Root and the catalpol in the Radix Rehmanniae medical material are also respectively as one of evaluation index.
Radix Puerariae, Radix Trichosanthis and Rhizoma Dioscoreae powder are broken into fritter, and Radix Rehmanniae is cut into small pieces.Take by weighing medical material (Radix Puerariae 10.6g, Radix Rehmanniae 6.36g, Radix Astragali 2.12g, Radix Trichosanthis 10.6g, Stigma Maydis 10.6g, Rhizoma Dioscoreae 1.06g) 6 parts, the water, 50% ethanol, 80% ethanol ultrasonic extraction that add respectively 8 times of crude drug amounts filtered after 20 minutes, filtrate concentrates and is settled in the 100ml measuring bottle, adopts the HPLC method to measure the peak area of puerarin in 6 duplicate samples, the results are shown in Table 8.Every kind of parallel two groups of tests of solvent.
Table 8 different solvents compares the extraction effect of puerarin
The result shows: the extraction effect of ethanol is better than water.
Consider that based on industrial feasibility investigated three kinds of extracting method commonly used: warm macerating method, ultrasonic extraction and heating and refluxing extraction method are to the extraction effect of puerarin.
Take by weighing 6 parts of medical materials (quantity is the same), add 8 times of amount 50% ethanol, reflux, extract, 1hr, supersound extraction 1hr, 50 ℃ of warm macerating extract 1hr respectively, filter, filtrate concentrates and is settled in the 250ml measuring bottle, after 0.45 μ m microporous filter membrane filters, adopt the HPLC method to measure the peak area of puerarin in 6 duplicate samples.The results are shown in Table 9.Every kind of parallel two groups of tests of method.
The investigation result of table 9 Different Extraction Method
Above result shows: the heating reflux method extraction effect is better, and suitable commercial production, so extracting method is selected heating reflux method.
Composition contained in the pharmaceutical composition crude drug of the present invention is relatively stable to heat, and therefore, according to top result of the test, selection ethanol is solvent, adopts the method for heating and refluxing extraction to extract.The major influence factors that heating reflux method extracts Effective Component of Chinese Medicine has: the consumption (B) of the concentration of ethanol (A), ethanol, extraction time (C) and extraction time (D) etc.So selected the orthogonal test table of 4 factors, 3 levels to test, the amount of the puerarin that obtains take extraction is screened best extraction process as evaluation index.The factor level table sees Table 10, orthogonal test table and the results are shown in Table 11.
Table 10 factor level table
Test method: take by weighing 9 parts of the 6 flavor medical materials such as Radix Puerariae, test according to each test of table 11, merge extractive liquid,, concentrating under reduced pressure, vacuum drying obtains extract.Precision takes by weighing this extract 25mg, places the 25ml measuring bottle, adds dissolve with methanol and is settled to scale, shakes up, and carries out HPLC and detect after 0.45 μ m microporous filter membrane filters.Adopt one point external standard method, puerarin must measure in the calculating medical material.The results are shown in Table 11.
As can be seen from Table 11, factor A (concentration of alcohol) is main influence factor, factor D (extraction time) also is one of major influence factors, not significantly difference between each level of factor B (ethanol consumption) and factor C (extraction time), be not major influence factors, but the impact of factor C is slightly larger than factor B.Therefore, be followed successively by A＞D＞C＞B take puerarin as the influence factor who investigates index.From factor A, A
3Be better than A
2And A
1, therefore select A
3From factor D, D
3Obviously be better than D
2And D
1, therefore select D
3Factor C selects C
3Factor B selects B
2So the preferred extraction process take puerarin as the investigation index is as A
3B
2C
3D
3
Table 11 orthogonal test table and result
Table 12 analysis of variance table
Must measure as investigating index take puerarin and to carry out variance analysis, the results are shown in Table 12.
The results of analysis of variance from table 12, factor A (concentration of alcohol) and factor D (extraction time) have the very significant impact to extraction effect, factor B (ethanol consumption) and factor C (extraction time) do not have the impact of significance on extraction effect, therefore consider that from the cost angle factor C selects C
1, consider intuitive analysis and the results of analysis of variance, determine that finally preferred extraction process is A
3B
2C
1D
3, namely medical material 50% alcohol heating reflux that adds 10 times of crude drug amounts extracts each 1 hour 3 times.
According to above optimal extract process, carry out 2 batches of process certification tests, the results are shown in Table 13.
The demonstration test result of table 13 extraction process
The result show this process stabilizing, reliable, repeatability is good.
Medicine of the present invention is a kind of compound preparation, and the synergism between medicine is the basis of its performance curative effect.So extraction process requires the various effective ingredient in its prescription medical material are all extracted as far as possible.In order to detect the extraction effect of above-mentioned optimum extraction process take puerarin as evaluation index, except measuring the content of puerarin, also select index composition astragaloside and catalpol in other two flavor medicine-Radixs Astragali and the Radix Rehmanniae, carry out TLC and detect.Developing solvent is CHCl
3: MeOH: H
2O (65: 35: 10, lower floor), 10% sulphuric acid-vanillin spraying colour developing.Can find out that by testing result the said extracted method also can effectively be extracted the Radix Astragali and Radix Rehmanniae medical material effective ingredient.
Simultaneously by orthogonal test, determine that 50% alcohol heating reflux that medical material adds 10 times of crude drug amounts extracts each 1 hour 3 times.Data by industrialized great production show that 10 times there is no significant difference with 8 times of crude drug amount solvent extraction effects, consider from large production cost, will extract multiplying power and make 8 times of solvent extractions into.
C, purification by macroporous resin research
The prescription medical material of medicine of the present invention is many, and the complicated component that contains in order to improve curative effect of medication, reduces dosage, should be as far as possible the impurity such as wherein monosaccharide, oligosaccharide, polysaccharide, starch, aminoacid, polypeptide, protein be removed.Macroporous adsorbent resin is a kind of polymeric sorbent that does not contain cation exchange groups, has the macroporous netlike structure.Have the following advantages: 1. the selectivity to Adsorption of Organic is good; 2. the stability of adsorbent is high, and mechanical strength is good, and is durable in use; 3. regeneration easily can Reusability; 4. applied range according to different purification requirements, is selected different resins.Because the adsorption of resin is physics chemical action, so that the material that is adsorbed more easily elutes from resin, resin itself is regenerated easily simultaneously, and macroporous adsorbent resin has been widely used in the purification of bioactive substance.Therefore, intend adopting Flavonoids by Macroporous Adsorption Resin to carry out purification.And measure the content of puerarin with the HPLC method, as evaluation index.
In macroporous adsorbent resin is investigated the maximum dynamic adsorption amount of puerarin, chosen macroporous adsorbent resin NKA commonly used, D201, D301, HPD400A, HPD100, HPD300 is (respectively by Tianjin sea light chemical industry company limited, TianXing, Bangbu resin company limited, the triumphant resin company limited of Shanghai strength, company's productions such as Cangzhou precious grace adsorbent resin material Science and Technology Ltd.) compare test, take by weighing respectively above-mentioned 6 kinds of each 20g of macroporous adsorbent resin that handled well, decompressing and extracting, put in the triangular flask, add respectively a certain amount of extract solution that obtains that extracts according to above-mentioned optimum extraction process, shake up soaked overnight.Next day, sucking filtration adopted the HPLC method to measure the peak area of puerarin in the filtrate, calculated each resin to the maximum static adsorbance of puerarin.The results are shown in Table 14.
The maximum static adsorbance of table 14 macroporous adsorbent resin is investigated the result
As can be seen from the above results, maximum static adsorbance HPD100 ≈ HPD300＞HPD400A＞D301＞NKA＞D201 selects HPD100, HPD300 and three kinds of resins of HPD400A to be used for following test.
Take by weighing respectively macroporous adsorbent resin HPD400A, HPD100, each 20g of HPD300 of having handled well, decompressing and extracting adds in the resin column of packing into after water mixes.Add solution (0.05g extract/ml water) 100ml according to the resulting extract of above-mentioned optimum extraction process in the resin column, the limit adds medicinal liquid, and the liquid that resin column flows out is below collected on the limit, the first two stream part each 25ml, rear 5 stream part each 10ml.Adopt HPLC method mensuration puerarin peak area wherein, until there is the puerarin chromatographic peak to detect, calculate the liquid volume that flows out this moment.Puerarin is more late to be detected, and illustrates that the reservation of puerarin on resin is better, and namely resin is larger to the maximum dynamic adsorption amount of puerarin.The results are shown in Table 15.
Result of the test shows that HPD400A model resin is collected in the liquid at the 7th part and detected puerarin; HPD300 model resin is collected in the liquid at the 4th part and is detected puerarin; HPD100 model resin is collected in the liquid at the 7th part and is detected puerarin, this shows that HPD400A and HPD100 model resin are larger to the maximum dynamic adsorption amount of puerarin, selects these two kinds of resins to be used for following test (seeing Fig. 1 and 2).
The investigation of the maximum dynamic adsorption amount of table 15 macroporous adsorbent resin
Annotate :-expression does not detect puerarin
Take by weighing respectively each 20g of macroporous adsorbent resin HPD400A, HPD100 that has handled well, decompressing and extracting adds in the resin column of packing into after water mixes.Add medicinal liquid (0.05g extract/ml water) 80ml according to the resulting extract of above-mentioned optimum extraction process in the resin column.Place after 30 minutes, use successively the H of 4 times of bed volumes
2O, 20%EtOH, 40%EtOH, 60%EtOH eluting merge respectively each gradient eluent, are evaporated to 100ml, adopt the HPLC method to measure the peak area of puerarin.The results are shown in Table 16 and Fig. 3.
As seen from Figure 3, water can not elute puerarin, and the puerarin that 20% ethanol elution gets off is maximum, and 40% ethanol just can elute most puerarin.Can determine thus, wash with water first, except large polar impurities such as desaccharide, then use 40% ethanol elution, the effective ingredient such as puerarin are eluted.The elution curve of these 2 kinds of resins is more close, all Radix Puerariae is have good separating effect, considers that the price comparison of HPD100 is cheap, and is a kind of macroporous adsorbent resin commonly used, therefore selects HPD100 model macroporous adsorbent resin to be used for following test.
The content of puerarin in each gradient eluent of table 16 macroporous adsorbent resin
Take by weighing HPD100 model macroporous adsorbent resin 20g, dress post after adding water and mixing adds medicinal liquid (0.05g extract/g water) 80ml according to the resulting extract of above-mentioned optimum extraction process in the resin column.After the standing adsorption 30 minutes, wash with water first, the eluent of each bed volume is collected 6 parts altogether as 1 stream part, and whether measure with the HPLC method has the puerarin chromatographic peak to detect; Then use 40% ethanol elution, collect the eluent of each column volume, collect altogether 6 parts, measure the puerarin peak area.The results are shown in Table 17.
The investigation result of table 17 elution volume
As can be seen from Table 17, during 6 times of bed volumes of water elution, can be washed till sugar-free.Puerarin mainly concentrates in front 2 parts of eluents of 40% ethanol elution, and puerarin content seldom after the 4th part of eluent.
By this test, can draw such eluting result: wash with water first to Molish reaction and be negative, then use 40% ethanol elution, collect the eluent of 3 times of column volumes.
In elution process, major influence factors comprises the blade diameter length ratio of applied sample amount (ratio of medical material and resin), liquor strength, elution flow rate, post bed etc., adopts orthogonal test, has investigated above-mentioned four principal elements to the impact of separating effect.The factor level table sees Table 18, and orthogonal test table sees Table 19.
Table 18 elution requirement factor level table
Table 19 eluting orthogonal test table and result of the test
Carry out intuitive analysis take puerarin amount/medical material amount as investigating index: as can be seen from Table 19, factor B (medical material and resin ratio) is than major influence factors; And factor A (resin column bed blade diameter length ratio), factor C (elution flow rate) and factor D (liquor strength) are not very large on the difference that affects of puerarin eluting.Optimum washing engaging condition can be chosen as A
2B
1C
2D
2
Table 20 analysis of variance table
Carry out variance analysis take puerarin amount/medical material amount as investigating index: as can be seen from Table 20, factor A (resin column bed blade diameter length ratio), factor B (medical material and resin ratio), factor C (elution flow rate) and factor D (liquor strength) all do not have the significance impact to the elute effect of puerarin.
Intuitive analysis and the results of analysis of variance all show do not have significant difference between each value of four factors.Factor B (medical material and resin ratio) considers, in non-overloading situation, should select as far as possible larger applied sample amount, so can select B
3Factor C (elution flow rate) is less on the impact of elute effect, in order to increase work efficiency, can select C
3D then selects D
2And optimum A is selected in the actual consideration of factor A from producing
2Therefore, optimum washing engaging condition can be chosen as A
2B
3C
3D
2
Using the purification with macroreticular resin technology, mainly is to remove the impurity such as some saccharides, reaches active constituent-enriched, reduces medication dose, improves the purpose of curative effect.In the process of using resin purification, should avoid the loss of effective ingredient as far as possible.Especially concerning compound recipe, it is especially important that this point seems.In utilizing the process of purification with macroreticular resin, the inventor is except adopting the HPLC method to follow the tracks of the eluting situation that detects puerarin, also adopt the method for TLC to detect the eluting situation of catalpol in astragaloside in the Milkvetch Root and the Radix Rehmanniae medical material, avoid the loss of effective ingredient as far as possible.
Adopt the TLC method, can find in 40% ethanol elution thing, to contain the compositions such as astragaloside and catalpol, illustrate that the effective ingredient major part is eluted.
Formulated the production technology of the 6 flavor medical materials such as Radix Puerariae according to above-mentioned result of the test, and carried out 2 batches of confirmatory experiments, the result all reaches the requirement of design, the results are shown in Table 21.
The demonstration test result of table 21 purifying process
Be the further efficient of checking 40% alcohol wash, after 40% alcohol wash, with 80% ethanol macroporous resin carried out eluting again, the TLC collection of illustrative plates of comparison paste-forming rate, puerarin content and other each compositions the results are shown in Table 22.Behind 40% ethanol elution, the rate of extract of 80% ethanol elution thing and puerarin content only are 10% of 40% ethanol elution part, and the TLC testing result shows that also 40% alcohol wash is substantially clean with the effective ingredient in Radix Puerariae, Radix Rehmanniae, the Radix Astragali, Radix Trichosanthis, Stigma Maydis and the Rhizoma Dioscoreae.
Table 2240% pure and mild 80% pure eluting result
D, drying process research
The Fructus Schisandrae Sphenantherae concentrated solution mixes with the concentrated solution of macroporous adsorbent resin 40% ethanol elution, adopts vacuum drying method dry, and temperature is 80 ℃, when vacuum is 0.09Mp.
E, granulation, tabletting, art for coating research
Granulate: determine to use fluidized granulating technique the good uniformity that the advantage of this technique is simply, supplementary product consumption is few, glibenclamide adds through Preliminary screening.The binding agents such as water, dextrin slurry, PVP K30 (being expressed as PVP in the table), carmethose (CMC-Na) have been compared in screening take yield rate, particle shape, particle size distribution, compressibility and disintegration etc. as performance assessment criteria, the results are shown in Table 23.
Table 23 binding agent the selection result
Illustrate: the computational methods of yield rate are that prepared 16～60 purpose particle weight are divided by inventory.PVP joins with 50% ethanol.The result shows, and is comparatively desirable take PVP as the binding agent resultant effect.
Mixed pressuring plate: disintegrating agent has been chosen 10% polyvinylpolypyrrolidone, 7% cross-linking sodium carboxymethyl cellulose, 15% amylum pregelatinisatum, 7% polyvinylpolypyrrolidone+3% carboxymethyl starch sodium and has been screened, comprehensively improve the effect of compressibility and disintegrate two aspects, take compressibility and disintegration as performance assessment criteria, the results are shown in Table 26.
Table 24 disintegrating agent the selection result
Comprehensively improve the effect of compressibility and disintegrate two aspects, select polyvinylpolypyrrolidone+carboxymethylstach sodium as disintegrating agent.
Coating: for improving outward appearance, improve patient's compliance, slice, thin piece is carried out coating, the clothing film also has certain moisture-proof role simultaneously, can increase the stability of tablet.Consider from the safety of production and the angle of environmental protection, adopt the aqueous coatings material.Selected green (85G61081) coating material of Shanghai Ka Lekang medical material company limited that pharmaceutical composition of the present invention is carried out coating, the disintegrate that this product is both economical, do not affect tablet has short, simple operation and other advantages of adhesive force height, unilateral light, preparation and simple to operate, environmental protection, man-hour.The lab scale test of the preparation method of embodiment 5 medicinal tablets of the present invention, pilot scale research and industrialization test manufacture
1. lab scale test
Press the technique of embodiment 1, carry out continuously 2 batches of (lot number is respectively: 06090702,06091801, Guangzhou Zhongyi Medicine Industry Co., Ltd) lab scale scale-ups.
Carried out the results are shown in Table 25 according to optimised process.
Table 25 lab scale result of the test
The result shows, process stabilizing, reliable, repeatability is good, can carry out scale-up.
2. pilot scale research
Press the technique of embodiment 1, carry out continuously three batches of pilot scales (lot number is 0701302,0701313,0701314, Guangzhou Zhongyi Medicine Industry Co., Ltd), every batch of medical material feeds intake by 40000, get the part extract powder and make respectively 10000, the result of study of three batches of pilot plant tests sees Table 26 and table 27.
The pilot scale research result of three batches of pharmaceutical compositions of the present invention of table 26
The quality examination result of three batches of pharmaceutical compositions of the present invention of table 27
Table 26 and table 27 result show that every quality index of three batches of pharmaceutical composition pilot scale amplification samples of the present invention is basically identical, illustrates that the formulation and technology of pharmaceutical composition of the present invention is reasonable, and favorable reproducibility can be used for amplifying and produces.
3. industrialization test manufacture
Press the technique of embodiment 1, carry out continuously 2 batches of (lot number is respectively: 20080701,20090501, Guangzhou Zhongyi Medicine Industry Co., Ltd) industrialization test manufactures.
Industrialization test manufacture research and pilot plant test result are contrasted concrete data such as following table:
Table 28 industrialization test manufacture and pilot scale research result
Three batches of pilot plant tests and two batches of test manufactures all are up to the standards by the touchstone of medicine of the present invention.As can be seen from Table 28, the extraction ratio of two batches of industrialization test manufactures (20080701,20090501) is higher than the data of three batches of pilot plant tests (0701302,0701313,0701314).In the test manufacture process, the technological parameter of test manufacture process is finished in strict accordance with the technological parameter of pilot plant test, and all finish at Same Site, equipment, technical process there is no abnormal conditions and occurs, infer that thus reason that industrialization test manufacture extraction ratio raises is because large than pilot plant test of the input amount of test manufacture, the production waste reduces.Result for the needs that adapt in the future industrialized great production and comprehensive test manufacture, pilot scale resets the sheet into 0.28g/ with sheet.
The pharmacodynamic study of embodiment 6 medicines of the present invention
1. medicine of the present invention is to the blood sugar reducing function research of anaphylactic type diabetes rat
Experiment material
Animal: select Wistar kind cleaning level male rat, body weight 210-230g is provided by Institute of Experimental Animals, Chinese Academy of Medical Sciences, licence numbering: SCXK-capital-2005-0003.Raise two grades of animal breeding plants, the quality certification number: moving word (capital) SYXK of doctor 2003-0008 number in the China TCM Academy of Sciences Xiyuan Hospital Experimental Animal Center.Room temperature: 22-25 ℃, relative humidity: 45-60%.
Feedstuff: product license number is provided by Beijing section Australia feed corporation,Ltd that pulls together: (joining) No. 238, operative norm GB14924,3-2001 are raised in the capital.Main nutrient composition: crude protein 〉=18%, crude fibre≤5%, coarse ash≤8%, calcium 0.8-1.8%, phosphorus 0.6-1.2%, lysine 〉=1.35%, Sal 0.5%, moisture≤10%, various electrolytes and minerals.
Medicine: pharmaceutical composition of the present invention (embodiment 1): 18g crude drug/g powder, lot number: 060630, the Peking University Chinese medicine Hyundai Research center, the Beijing Huayi Shennong Medicine Technology Co., Ltd provides.Metformin hydrochloride tablet: the 0.25g/ sheet, lot number: 060505, the Beijing XieHe medicine Factory produces, the accurate word H11020976 of traditional Chinese medicines.Glyburide: lot number 060404 is provided by Guangzhou Zhongyi Medicine Industry Co., Ltd.Diabetes pill (former pharmaceutical composition): 4.346g crude drug/g medicine, lot number: 060405, Guangzhou Zhongyi Medicine Industry Co., Ltd produces, the accurate word Z44020045 of traditional Chinese medicines.
Reagent: citric acid and sodium citrate: the Beijing Chemical Plant produces, lot number: 960904.Streptozotocin (STZ): Sigma company produces, lot number: S0130, biological medical science and technology company limited packing over the sky, Beijing.Blood sugar test paper: German Roche Holding Ag produces, lot number: 22939331, Luo Shi diagnostic products (Shanghai) Co., Ltd. packing.Insulin radioimmunological kit: the Ke Meidongya of Beijing Bioisystech Co., Ltd, lot number: 20061225.Glucagon radioimmunological kit: the Ke Meidongya of Beijing Bioisystech Co., Ltd, lot number: 20061225.Glycolated hemoglobin: German Roche Holding Ag produces, lot number: 200612.
Experimental technique
Wistar kind cleaning level male rat is selected in experiment, body weight 210-230g, each group is all used streptozotocin (STZ) tail vein injection (42mg/kg except the blank group, 0.1M the citrate buffer solution preparation), the tail point is got the hematometry fasting glucose after 72 hours, with greater than 16.7mmol/l as model success rat.Grouping: (1) blank group; (2) model group; (3) positive drug metformin group 0.2g/kg; (4) glyburide group 1.38mg/kg; That is: contained glyburide amount among former dosage form 6g crude drug/kg; (5) diabetes pill group 6g crude drug/kg (per 5 balls conversion contains crude drug amount 5.43g); (6) the heavy dose of group of the medicine in the embodiment of the invention 1 12g crude drug/kg (every conversion contains crude drug amount 5.43g); (7) dosage group 6g crude drug/kg in the medicine of the present invention; (8) medicine small dose group 3g crude drug/kg of the present invention.Each organizes all according to dosage gastric infusion except the blank group.Survey weekly body weight and ingest, adjust dose according to body weight, the tail point is got blood and is surveyed blood glucose every other week.Successive administration two months, ventral aorta is got whole blood (anticoagulant of 25ulEDTA+1ml whole blood) mensuration glycolated hemoglobin, gets blood 1ml separation of serum mensuration insulin, is got blood 1ml separated plasma (anticoagulant of 25ulEDTA+1ml whole blood) mensuration glucagon after the Animal Anesthesia, gets whole blood 3ml (anticoagulant heparin) and measures the rat serum rheology.Get pancreas and make pathology (HE dyeing and aldehyde-fuchsin spy dye).The result carries out statistical procedures.
Experimental result
Rat through streptozotocin (STZ) tail vein injection after 72 hours the tail point get the hematometry fasting glucose, each group obviously raises with matched group comparison blood glucose before the medicine, the average blood sugar level illustrate that modeling is successful about 20mmol/l.Each organizes continuous gastric infusion two months, and diabetes pill group, the big or middle dosage group of medicine of the present invention and model group relatively have blood sugar reducing function during 4～8 week, and small dose group has blood sugar reducing function during 6～8 week, and heavy dose of group blood sugar lowering rate reaches more than 20%.The results are shown in Table 29.
Table 29 different pharmaceutical is on the impact of anaphylactic type blood glucose in diabetic rats
Compare with matched group: * * * P＜0.001; With model group relatively: #P＜0.05 ##P＜0.01 ###P＜0.001 (small dose group during 6 week dead 1)
Each organizes continuous gastric infusion two months, each dosage group of diabetes pill and medicine of the present invention and model group are relatively, glycolated hemoglobin (HBAlc) and glycolated hemoglobin percentage ratio (Halc%) descend to some extent, but no difference of science of statistics, hemoglobin (HB) compares no significant difference with model group.The results are shown in Table 30.
Table 30 different pharmaceutical is on the impact of the indexs such as anaphylactic type diabetes rat glycolated hemoglobin
Compare with matched group: * * * P＜0.001; Compare with model group: #P＜0.05
The continuous gastric infusion of each dosage group of diabetes pill and pharmaceutical composition of the present invention two months, the heavy dose of group of new Xiaoke Tablets and metformin group and model group are relatively, glucagon descends (P＜0.05) to some extent, and insulin level has the trend of increasing, but not statistically significant.Small dose group is showed no obvious impact to insulin and glucagon in glyburide, former dosage form and the new Xiaoke Tablets, the results are shown in Table 31.
Table 31 different pharmaceutical is on the impact of anaphylactic type diabetes rat blood insulin, glucagon
Compare with matched group: * * * P＜0.001; Compare with model group: #P＜0.05
The continuous gastric infusion of each treated animal two months, the new big or middle dosage group of Xiaoke Tablets and former dosage form group and model group comparison, body weight increase obviously that (P＜0.01-0.001), polydipsia polyuria symptom obviously alleviates.
Table 32 different pharmaceutical is on the impact (gram) of anaphylactic type diabetes rat body weight
Compare with matched group: compare with model group * * * P＜0.001: #P＜0.05 ##P＜0.01 ##P＜0.01
Table 33 different pharmaceutical is on the impact (gram) of anaphylactic type diabetes rat body weight
Compare with matched group: compare with model group * * * P＜0.001: #P＜0.05 ##P＜0.01 ###P＜0.01
The continuous gastric infusion of each treated animal two months, each dosage group of medicine of the present invention and diabetes pill group and model group relatively, food ration has in various degree decline, and (P＜0.05-0.001), along with the reduction of blood sugar level, the symptom of diabetes obviously alleviates.The results are shown in Table 34, table 35.
Table 34 different pharmaceutical on the impact (gram) of anaphylactic type diabetes rat food ration (n=2,
)
Compare with matched group: * P＜0.05 * * P＜0.01 * * * P＜0.001; Compare with model group: #P＜0.05 ##P＜0.01
Table 35 different pharmaceutical is on the impact (gram) of anaphylactic type diabetes rat food ration
Compare with matched group: * P＜0.05 * * P＜0.01 * * * P＜0.001; Compare with model group: #P＜0.05 ##P＜0.01 ###P＜0.01
Medicine effect of the present invention is learned result of study and is shown that medicine of the present invention has blood sugar reducing function to diabetes rat, and suitable with the diabetes pill blood sugar reducing function; The simultaneously test of acute toxicity and long term toxicity shows that the safe dose of medicine of the present invention is larger, is safe in the dose scope, and does not find that therebetween the animal untoward reaction occurs.Thereby definite pharmaceutical composition of the present invention is safely and effectively.In sum, medicine of the present invention has obvious blood sugar reducing function to the anaphylactic type diabetes rat, and is basic identical with the diabetes pill effect under the Isodose, namely takes pharmaceutical composition a slice of the present invention and is equivalent to take 5 of diabetes pilles.
The present invention has also carried out to the diabetes rat carbohydrate tolerance and to the research of diabetic mice blood sugar reducing function, the result shows: medicine of the present invention to the diabetes rat carbohydrate tolerance improve significantly, under the same dose, medicine of the present invention has blood sugar reducing function to diabetic mice, basically identical with the diabetes pill blood sugar reducing function, take pharmaceutical composition a slice of the present invention and be equivalent to take 5 of diabetes pilles.Concrete data are omitted.
More than be for the specifying of possible embodiments of the present invention, but this embodiment limits claim of the present invention, allly do not break away from equivalence of the present invention and implement or change, all should be contained in the claim of the present invention.
Claims (5)
1. the preparation method of a Remedies for diabetes, the active component of this pharmaceutical composition is to be made by the crude drug of following ratio: Radix Rehmanniae 600-1200 weight portion, Radix Astragali 200-400 weight portion, Rhizoma Dioscoreae 100-200 weight portion, Radix Puerariae 1000-2000 weight portion, Radix Trichosanthis 1000-2000 weight portion, Stigma Maydis 1000-2000 weight portion, Fructus Schisandrae Sphenantherae 200-400 weight portion, glibenclamide 0.2-1.5 weight portion, it is characterized in that, may further comprise the steps:
(1) get the Fructus Schisandrae Sphenantherae medicinal material coarse powder, extract 1～3 time with 60～95% alcohol heating reflux of 4～8 times of crude drug amounts, each 1～3 hour, crossing the concentrated relative density that obtains of leaching filtrate decompression was 1.05～1.2 concentrated solution 1;
(2) get Radix Rehmanniae, the Radix Astragali, Rhizoma Dioscoreae, Radix Puerariae, Radix Trichosanthis, Stigma Maydis, extract 1～3 time with the 50-70% alcohol heating reflux of 6～12 times of crude drug amounts, each 1～2 hour, it was 1.05～1.2 concentrated solution 2 that the extracting solution concentrating under reduced pressure obtains relative density;
(3) concentrated solution 2 is injected in the HPD100 or HPD400A macroporous adsorptive resins that has handled well, first water is that eluent is washed till the Molish reaction and is negative, and discards eluent; With the 40-60% ethanol elution of 2-4 times of cylinder accumulated amount, collect ethanol elution again, Recycled ethanol and the concentrated concentrated solution 3 that obtains relative density 1.05～1.2;
(4) with concentrated solution 3 and concentrated solution 1 mix homogeneously, drying is pulverized granulation;
(5) add glibenclamide and various conventional adjuvant, method of Chinese medicinal prepares and get final product routinely.
2. the preparation method of described medicine according to claim 1 is characterized in that described medicine is tablet, capsule or pill.
3. the preparation method of described medicine according to claim 1 is characterized in that, described step (1) is for the 80% alcohol heating reflux extraction of 6 times of crude drug amounts 2 times, and each 2 hours, the relative density of described concentrated solution 1 was 1.10～1.15.
4. the preparation method of described medicine according to claim 1 is characterized in that, described step (2) is for the 50% alcohol heating reflux extraction of 8 times of crude drug amounts 3 times, and each 1 hour, the relative density of described concentrated solution 2 was 1.05～1.10.
5. the preparation method of described medicine according to claim 1, it is characterized in that the technological parameter of macroporous adsorbent resin is described in the step (3): medical material: resin is 1: 1; Absorption flow velocity and elution flow rate be 3 times of column volumes/hour; The blade diameter length ratio of post bed is 1: 3; The concentration of ethanol is 40%, and consumption is 3 times of cylinder accumulated amounts.
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| CN1861176A (en) * | 2005-05-18 | 2006-11-15 | 广州中一药业有限公司 | Medicinal composition for treating diabets, and its prepn. method |
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| CN1861176A (en) * | 2005-05-18 | 2006-11-15 | 广州中一药业有限公司 | Medicinal composition for treating diabets, and its prepn. method |
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