CN100486637C

CN100486637C - Composition of external remedy for treating hemorrhoids, and preparation method

Info

Publication number: CN100486637C
Application number: CNB2006100757125A
Authority: CN
Inventors: 张毅; 陈永泰
Original assignee: Individual
Current assignee: Individual
Priority date: 2006-04-18
Filing date: 2006-04-18
Publication date: 2009-05-13
Anticipated expiration: 2026-04-18
Also published as: CN1872326A

Abstract

An exterior-applied Chinese medicine for treating pile and the local swelling, pain and bleeding of anus caused by operation is prepared from coptis root, China gall and Japanese ampelopsis root through pulverizing coptis root, respective decocting others, filtering, concentrating, mixing all together and pulverizing.

Description

A kind of externally-applied medicinal composition for the treatment of hemorrhoid and preparation method thereof

Technical field

The present invention relates to a kind of pharmaceutical composition and preparation method thereof and method of quality control, particularly a kind of local swelling that hemorrhoid and anal operation of postop. occur, pain, hemorrhage externally-applied medicinal composition and preparation method thereof and method of quality control for the treatment of.

Background technology

Hemorrhoid are under the rectum end mucosa and the hemorrhoidal veins silk under the anal canal skin enlarges, the formed softish vein of varicose group.Be divided into internal hemorrhoid, external hemorrhoid and mixed hemorrhoid clinically.Hemorrhoid are common, multiple diseases.The report of domestic generaI investigation shows: the sickness rate of hemorrhoid reaches 51.96%, and the adult accounts for 70%.Hemorrhoid sickness rate abroad also reaches 56.7%.Theory of Chinese medical science thinks that the morbidity of hemorrhoid is an imbalance of YIN and YANG, and the internal organs QI and blood is deficient, humidification, heat, wind, effect such as dry again, and feelings will internal injury, and influences such as daily life occupation cause disorder of QI and blood, blockage of meridian, congestion foul smell is made a bet, and the poly-anus of knot dashes to send out and is hemorrhoid.

A lot of to the treatment of hemorrhoid method both at home and abroad at present.Drug therapy (as oral, the drug of topical application, turunda, fumigation and wash method etc.) is arranged, new and high technology (as laser, microwave, infrared ray, electronics etc.), injection treatment, operative therapy etc.Because of restrictions of having ready conditions such as new and high technology, injection treatment, operative therapys, so most hemorrhoid patients still needs Drug therapy.Western medicine such as antibiotic commonly used, hemostasis, analgesic.In the treatment of internal hemorrhoid, injection treatment used nearly 100 years, and is effective to early-stage cases and hemostasis, but invalid to serious internal hemorrhoid in late period.The effect of infra-red coagulation therapy is similar to injection treatment, but the danger of secondary hemorrhage is arranged.The Barron ligation method is easy, and idol has pain, edema and hemorrhage, but the relapse rate height.Because the pathological factor of relevant hemorrhoid is not clear.The doctor trained in Western medicine conservative treatment is got symptomatic treatment more, as takes pain relieving, and medicines such as hemostasis, antiinflammatory are with relief of symptoms.Chinese herb decoction opinion treatment internal hemorrhoid has obtained comparatively effectively curative effect, but the differentiation of symptoms and signs for classification of syndrome confusion is difficult to repeated authentication, and lacks unified diagnostic criteria, is unfavorable for the standardization for the treatment of.Curative effect from present clinical treatment hemorrhoid, better with special Fang Weizhu with the effect of disease plus-minus, but dosage form is restricted in the use, is unfavorable for large-scale long-term treatment, therefore the efficient medicine of screening is made the easy one-tenth side's patent medicine of dosage form on special side basis, is clinical needs.

Summary of the invention

The object of the invention is to provide a kind of externally-applied medicinal composition and preparation method thereof, and another purpose of the present invention is to provide the method for quality control and the purposes of this pharmaceutical composition.

The present invention seeks to be achieved through the following technical solutions:

The crude drug of pharmaceutical composition of the present invention consists of: Rhizoma Coptidis 150-280 weight portion Radix Ampelopsis 150-280 weight portion Galla Chinensis 350-700 weight portion Radix Curcumae 300-550 weight portion Olibanum 40-65 weight portion.

The preferred weight proportioning of above-mentioned raw materials medicine is as follows:

Rhizoma Coptidis 212 weight portion Radix Ampelopsis 231.6 weight portion Galla Chinensiss 565.5 weight portion Radix Curcumaes 463 weight portion Olibanums 53 weight portions; Rhizoma Coptidis 160 weight portion Radix Ampelopsis 170 weight portion Galla Chinensiss, 360 weight portion Radix Curcumae 540 weight portion Olibanums, 60 weight portions or Rhizoma Coptidis 270 weight portion Radix Ampelopsis 260 weight portion Galla Chinensiss 690 weight portion Radix Curcumaes 310 weight portion Olibanums 45 weight portions.

The invention described above pharmaceutical composition can also add the Borneolum Syntheticum of following weight portion and be made up of the Six-element crude drug: Borneolum Syntheticum 15-30 weight portion; The preferred weight proportioning is respectively: Borneolum Syntheticum 21 weight portions; Borneolum Syntheticum 27 weight portions or Borneolum Syntheticum 16 weight portions.

Pharmaceutical composition crude drug of the present invention adds conventional adjuvant, according to common process, makes external preparation such as clinical acceptable suppository, ointment, fumigation and washing agent or effervescent tablet.

Pharmaceutical composition of the present invention can also add the semi-synthetic fatty acid ester of following weight portion and make suppository: semi-synthetic fatty acid ester 1200-1500 weight portion; The preferred weight proportioning is as follows: semi-synthetic fatty acid ester 1286 weight portions; Semi-synthetic fatty acid ester 1213 weight portions or semi-synthetic fatty acid ester 1428 weight portions.

The preparation technology of pharmaceutical composition suppository of the present invention is as follows:

Get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; Radix Curcumae, Radix Ampelopsis add the decocting that 4-8 doubly measures and boil 2-4 time, the each decoction 1-2 hour, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.08-1.12 clear paste that medicinal liquid is concentrated into 60 ℃ of relative densities; Galla Chinensis adds 5-9 times of water gaging and decocts 2-4 time, decocted 0.5-1 hour at every turn, and collecting decoction, filtration, it is 1.08-1.12 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste; Be 150-160 ℃ in inlet temperature, spray drying under 80-90 ℃ of conditions of leaving air temp is mixed with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester, place evaporating dish, 40 ± 2 ℃ of fusings of water-bath add above-mentioned fine powder, add Borneolum Syntheticum again, and mix homogeneously pours in the bolt mould, cooling, and suppository is made in the demoulding.

The preferred for preparation method of pharmaceutical composition suppository of the present invention is as follows:

Get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; The decocting that Radix Curcumae, Radix Ampelopsis add 6 times of amounts boils 3 times, the each decoction 1.5 hours, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.10 clear paste that medicinal liquid is concentrated into 60 ℃ of relative densities; Galla Chinensis adds 7 times of water gagings and decocts 3 times, decocts 0.5 hour at every turn, and collecting decoction filters, it is 1.10 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste, is 155 ℃ in inlet temperature, spray drying under 85 ℃ of conditions of leaving air temp is mixed with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester, place evaporating dish, water-bath is dissolved for 40 ℃, adds above-mentioned fine powder, adds the Borneolum Syntheticum mix homogeneously again, pour in the bolt mould, and cooling, the demoulding, suppository is made in packing.

The method of quality control of pharmaceutical composition of the present invention comprises following discriminating and/or assay:

Discriminating comprises one or more in the following method:

A, get suppository 2g of the present invention, grind, add 60-90 ℃ of petroleum ether 15-25ml, supersound process 4-6 minute, filter, medicinal residues are waved most solvent, add methanol 15-25ml, and supersound process 10-20 minute, filter, filtrate is as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 2.5-7.5ml, and supersound process 10-20 minute, filter, filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.3-0.7mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=4-10:1:1-3 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get suppository 4g of the present invention, add water 30-50ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1-2, uses ether extraction 1-3 time, each 15-25ml, merge ether solution, volatilize, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=4-8:3-5:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay in the method for quality control is as follows:

Chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=40-50:15-25:25-30 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000; The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get this pharmaceutical composition suppository under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=75-125:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=75-125:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 10-20 μ l of drawing, inject chromatograph of liquid, measure, promptly; The every gram of pharmaceutical composition suppository of the present invention contains Rhizoma Coptidis with berberine hydrochloride C ₂₀H ₁₈ClNO ₄Meter must not be less than 6mg.

The method of quality control of pharmaceutical composition of the present invention is preferably as follows to be differentiated and/or assay:

Discriminating comprises one or more in the following method:

A, get suppository 2g of the present invention, grind, add 75 ℃ of petroleum ether 20ml, supersound process 5 minutes filters, and medicinal residues are waved most solvent, add methanol 20ml, and supersound process 15 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=7:1:2 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get suppository 4g of the present invention, add water 40ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1.5, and with ether extraction 2 times, each 20ml merges ether solution, volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=6:4:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay: chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=50:20:30 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000; The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get the suppository 20g of the present invention under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=100:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=100:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and the need testing solution 15 μ l of drawing, inject chromatograph of liquid, measure, promptly; The every gram of suppository of the present invention contains Rhizoma Coptidis with berberine hydrochloride C ₂₀H ₁₈ClNO ₄Meter must not be less than 6mg.

This pharmaceutical composition with the Radix Ampelopsis bitter cold let out clearly, the hot detumescence of loosing, heat-clearing and toxic substances removing, eliminating carbuncle eliminating stagnation; Rhizoma Coptidis heat clearing and damp drying, eliminating fire and detoxication; The Olibanum promoting of the circulation of QI and removing the obstruction in the collaterals, promoting blood circulation to remove blood stasis, reducing swelling and alleviating pain, gas is capable then seek defend unimpeded, battalion defend unobstructed then evil do not have be detained, making becomes silted up goes, swollenly disappear, pain is ended; The Radix Curcumae acrid in the mouth can loose and can manage it, and can invigorate blood circulation, again can promoting QI circulation for relieving depression and reach the effect of pain relieving, and simultaneously arduous and cold in nature, can pleasant pathogenic fire reducing and cooling blood for hemostasis; Radix Curcumae is joined Galla Chinensis, and the power of having silt when astringent therapy is astringent or styptic treatment for spontaneous sweating makes blood only not have the fraud of staying silt; Borneolum Syntheticum fragrance, the evil that exorcises evil spirits, the rheumatism of loosing; All medicines share, and gather the merit of heat clearing away, hemostasis, antiinflammatory, pain relieving altogether.Pharmacodynamic experiment shows: 1. this pharmaceutical composition suppository has tangible antiinflammatory action.Can obviously suppress rat paw edema that Ovum Gallus domesticus album causes and the mice auricle swelling that causes by dimethylbenzene; Suppressing the mouse peritoneal capillary permeability increases.2. this pharmaceutical composition suppository has certain analgesic activity.Can obviously improve the hot plate pain threshold of mice; Can obviously reduce the writhing response number of times that acetic acid brings out mice.3. this pharmaceutical composition suppository has certain anastalsis.Can shorten the mice bleeding time, clotting time and the tail point local hemostasis time.4. this pharmaceutical composition suppository has antibacterial activity to the common Gram-positive of clinical separation, negative pathogenic bacterium.Its minimum inhibitory concentration MIC value scope is 0.125-125mg crude drug/ml.Acute toxicity testing shows: this pharmaceutical composition suppository has very big safety range.The local irritation experimentation shows: this pharmaceutical composition suppository under the dosage of 0.17g crude drug in whole/kilogram to the effect of tame rabbit rectum nonirritant.

Method of quality control of the present invention shows its good reproducibility according to experimental result, and instrument precision is good; This pharmaceutical composition stability and repeatability are good, higher, the separator well of supersound extraction efficient.The extraction process condition is basicly stable, tool repeatability.Technology quantity-produced good stability, process conditions are rationally feasible.Simultaneously pharmaceutical composition suppository of the present invention can reach the ideal effect of performance general action with correct rectally method, and it is with conspicuous characteristics with oral comparison: the one, and rectum absorbs the first pass effect that can avoid liver, reduces the toxic and side effects to liver.The 2nd, medicine is not subjected to the influence of gastrointestinal pH, enzyme, and it is little to absorb more oral interference at rectum.The 3rd, rectally fast, effectively, patient and the old man and the child of the medicine that is suitable for can not or being reluctant swallowing.And suppository uses, carries, transports all very convenient.

Following experimental example is used to further specify but is not limited to the present invention.

The antiinflammatory experiment of 1 pharmaceutical composition suppository of experimental example

1. this pharmaceutical composition suppository causes the influence of rat paw edema to Ovum Gallus domesticus album

Get 60 of body weight 130-170g healthy SD rats, experiment prospective adaptation raising 3 days, rat is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), prednisone positive controls (3.23mg/kg) and six groups of excipient group, every group 10, male and female half and half.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 2.0 milliliters/100g body weight equal volume, and the normal saline matched group is given isopyknic normal saline, and every rat of excipient group is given substrate (0.26g substrate/kilogram).Clamp the rat anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.Every day 1 time, successive administration 7 days.1h causes inflammation in every rat left foot palm subcutaneous injection 10% fresh albumen 0.05ml after the last administration.And it is long-pending to survey the Mus corpus unguis with drainage behind 30min, 60min, 120min, the 240min behind the Yu Zhiyan, and causes scorching front volume and compares.So that scorching back volume deducts and causes scorching front volume calculating swelling degree, and data are carried out statistical analysis.See Table 1.

This pharmaceutical composition of table 1 suppository is to the influence of rat paw edema due to the Ovum Gallus domesticus album

Experimental result shows: this pharmaceutical composition suppository can obviously suppress the rat paw edema that Ovum Gallus domesticus album causes.This pharmaceutical composition suppository height, middle dosage group can make rat 30min, 60min, 120min swelling degree significantly reduce (P＜0.001) than the normal saline matched group, and low dose of this pharmaceutical composition suppository can make rat 30min, 60min, 120min swelling degree than matched group significantly descend (P＜0.05).The high, medium and low dosage group of this pharmaceutical composition suppository can make rat 240min swelling degree than matched group significantly descend (P＜0.05).The excipient effect is similar to normal saline, fails inflammation-inhibiting reaction (P〉0.05).Show that this pharmaceutical composition suppository can suppress experimental early stage development of inflammation, promotes it to disappear.

2. this pharmaceutical composition suppository influence that the mouse peritoneal capillary permeability is increased, get 60 of the healthy KM mices of body weight 18-22g, experiment prospective adaptation raising 3 days, mice is divided into the normal saline matched group at random, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram), dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), six groups of indometacin positive controls (11.36mg/kg) and excipient group (0.36g substrate/kilogram), every group 10, male and female half and half.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 0.3 milliliter/10g body weight equal volume, and the normal saline matched group is given isopyknic normal saline, and every mice of excipient group is given substrate (0.36g/ kilogram).Clamp the mice anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.Every day 1 time, successive administration 7 days.1h after the last administration, after every mice was injected the blue normal saline solution 0.1ml/10g of 0.5% she Wen body weight by the tail vein, lumbar injection 0.6% glacial acetic acid normal saline solution 0.2ml/ only immediately.Take off neck after 20 minutes and put to death animal, cut off skin of abdomen muscle, divide with the 6ml normal saline and wash the abdominal cavity for several times, suction pipe sucking-off cleaning mixture merges the back and adds normal saline to 10ml, and the centrifugal 15min of 3000rpm gets supernatant to 590nm colorimetric determination OD value.The data of gained are carried out statistical analysis.See Table 2.

This pharmaceutical composition of table 2 suppository is to the influence of mouse peritoneal capillary permeability (X ± S)

Annotate: compare " * " P＜0.05 with matched group; " * * " P＜0.01; " * * * " P＜0.001

Experimental result shows: this pharmaceutical composition suppository height, middle dosage group mouse peritoneal cleaning mixture show that than the remarkable decline of normal saline matched group abdominal cavity cleaning mixture OD value (P＜0.01, P＜0.05) it can obviously suppress H ⁺Due to the increasing of mouse peritoneal capillary permeability.Excipient group mouse peritoneal cleaning mixture then close (P〉0.05) with normal saline matched group abdominal cavity cleaning mixture OD value.

3. this pharmaceutical composition suppository is to the influence of mice by auricle edema due to the dimethylbenzene

Get 60 of the healthy KM mices of body weight 18-22g, experiment prospective adaptation raising 3 days, mice is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), indometacin positive controls (11.36mg/kg) and six groups of excipient group (0.36g/ kilogram), every group 10, male and female half and half.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 0.3 milliliter/10g body weight equal volume, and the normal saline matched group is given isopyknic normal saline, and every mice of excipient group is given substrate (0.36g/ kilogram).Clamp the mice anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.Every day 1 time, successive administration 7 days.30min after the last administration drips dimethylbenzene in mouse right ear and causes inflammation, puts to death mice behind the 15min, punch in the identical part of mice left and right sides ears with card punch, and the both sides auricle of weighing respectively, the difference of two auricle weight is the swelling degree, calculates swelling rate and inhibitory rate of intumesce.See Table 3.

This pharmaceutical composition of table 3 suppository to mice by the influence of auricle edema due to the dimethylbenzene (X ± S)

Experimental result shows: this pharmaceutical composition suppository can obviously suppress the mice auricle swelling that caused by dimethylbenzene.Wherein the strongest with the high dose group effect, significantly reduce (P＜0.01) than normal saline matched group swelling degree.In this pharmaceutical composition suppository, low dose group swelling degree also has remarkable reduction (P＜0.05) than the normal saline matched group.And excipient group swelling degree is compared no significant difference (P〉0.05) with the normal saline matched group.

The analgesic experiment of 2 pharmaceutical composition suppositorys of experimental example

1. this pharmaceutical composition suppository makes its temperature remain on (55 ± 0.5) ° C to the influence adjusting hot-plate instrument of mice hot plate reaction, it is some to get body weight 18-22g Healthy female KM mice, each 1 is placed on the hot plate, mice is from being placed on the hot plate to the pain thresholding of required time of metapedes (second) as this Mus occurring licking, allly licks the metapedes time less than 5 seconds or give it up greater than 30 seconds or the person of jumping over.Filter out 60 mices, mice is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), codeine phosphate positive controls (13.64mg/kg) and six groups of excipient group (0.36g/ kilogram), 10 every group.Repeat to survey the normal pain thresholding of respectively organizing mice, get twice meansigma methods and be worth before as medicine.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 0.3 milliliter/10g body weight equal volume, the normal saline matched group is given isopyknic normal saline, every mice of excipient group is given substrate (0.36g/ kilogram), clamp the mice anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.The codeine phosphate gastric infusion.Every day 1 time, successive administration 7 days.30min, 60min, 90min measure the pain thresholding of each treated animal after the last administration.The data of gained are carried out statistical analysis.See Table 4.

This pharmaceutical composition of table 4 suppository causes the influence (X ± S) in mice pain territory response time to hot plate method

Experimental result shows: compared remarkable increase (P＜0.01) before 30min pain territory and the administration behind this pharmaceutical composition suppository high dose group medicine, comparing before 30min pain territory and the administration behind the dosage group medicine in this pharmaceutical composition suppository also has remarkable increase (P＜0.05).Compared remarkable increase (P＜0.05) before 60min pain territory and the administration behind this pharmaceutical composition suppository height, the middle dosage group medicine.Show this pharmaceutical composition suppository to mice because the pain reaction that causes of thermostimulation has analgesic activity.

2. this pharmaceutical composition suppository is to the influence of mice acetic acid twisting reaction

Get 60 of the healthy KM mices of body weight 18-22g, experiment prospective adaptation raising 3 days, mice is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), codeine phosphate positive controls (13.64mg/kg) and six groups of excipient group (0.36g/ kilogram), every group 10, male and female half and half.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 0.3 milliliter/10g body weight equal volume, the normal saline matched group is given isopyknic normal saline, every mice of excipient group is given substrate (0.36g/ kilogram), clamp the mice anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.The codeine phosphate gastric infusion.Every day 1 time, successive administration 7 days.1h after the last administration, every mouse peritoneal is only injected 0.5% acetic acid 0.2ml/.Observe every writhing response (abdominal part indent, stretching, extension hind leg, buttocks are raised) number of times that mice occurs in 15 minutes, calculate and respectively organize the writhing response suppression ratio.See Table 5.

The influence that this pharmaceutical composition of table 5 suppository reacts the mice acetic acid twisting (X ± S)

Experimental result shows: this pharmaceutical composition suppository can obviously reduce by the mouse writhing reaction times due to the acetic acid.Wherein the strongest with the high dose group effect, turn round the body number of times than the normal saline control group mice and significantly reduce (P＜0.01).In this pharmaceutical composition suppository, low dose group mouse writhing number of times also has remarkable reduction (P＜0.05) than the normal saline matched group.And excipient group swelling degree is compared no significant difference (P〉0.05) with the normal saline matched group.Show that this pharmaceutical composition suppository has certain inhibitory action to the pain reaction of the mice due to the chemical stimulation.

The hemostasis experiment of 3 pharmaceutical composition suppositorys of experimental example

1. this pharmaceutical composition suppository is got 60 of the healthy KM mices of body weight 18-22g to the influence that mice goes out clotting time, experiment prospective adaptation raising 3 days, mice is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), YUNNAN BAIYAO positive controls (0.30g/kg) and six groups of excipient group (0.36g/ kilogram), every group 10, male and female half and half.The administration group is irritated drug solution with above-mentioned dosage respectively to rectum with 0.3 milliliter/10g body weight equal volume, and the normal saline matched group is given isopyknic normal saline, and every mice of excipient group is given substrate (0.36g/ kilogram).Clamp the mice anus in case medicine is excessive with Small clamp after the administration, take off behind the 1h.Every day 1 time, successive administration 7 days.Behind last administration 1h, capillary glass-tube with internal diameter 1mm inserts the blood sampling of mouse orbit angular vein clump, the blood post reaches 5cm to the capillary glass-tube, and then every 30s 1 trifle that fractures, record is from the time (clotting time) of blood sampling when the blood clotting silk appears in the capillary glass-tube that fractures; Simultaneously, dock, inhale to dehematize with filter paper every 30s and drip the time (bleeding time) of record when docking extremely is drawn to depletion of blood with filter paper 1 time apart from tail point 6mm place.Data are carried out statistical analysis.See Table 6.

This pharmaceutical composition of table 6 suppository to mice go out, the influence of clotting time (X ± S)

Experimental result shows: this pharmaceutical composition suppository high dose can make the mice bleeding time shorten, and it is compared with the normal saline matched group, and there were significant differences (P＜0.01).Simultaneously, this pharmaceutical composition suppository height, middle dosage all can shorten the clotting time (P＜0.01, P＜0.05) of mice.Show that this pharmaceutical composition suppository has significantly anastalsis to mice body inner model.

2. this pharmaceutical composition suppository is to the local hemostatic influence of mouse tail point

Get 60 of the healthy KM mices of body weight 18-22g, experiment prospective adaptation raising 3 days, mice is divided into normal saline matched group, this pharmaceutical composition suppository high dose group (1.00g crude drug in whole/kilogram) at random, dosage group in this pharmaceutical composition suppository (0.50g crude drug in whole/kilogram), this pharmaceutical composition suppository low dose group (0.25g crude drug in whole/kilogram), YUNNAN BAIYAO positive controls (0.30mg/kg) and six groups of excipient group (0.36g/ kilogram), every group 10, male and female half and half.During experiment mice is put into holder, sentence blade at distance tail point 1.0cm and block, immerse immediately in 37 ° of C normal saline or the different medicinal liquid, pick up counting immediately, observed once every 30 seconds behind the 5min, till no longer hemorrhage, be its bleeding time, the result carries out statistical analysis.See Table 7.

This pharmaceutical composition of table 7 suppository is to the local hemostatic influence of mouse tail point (X ± S)

Experimental result shows: this pharmaceutical composition suppository height, middle dosage all can shorten the time of mouse tail point local hemorrhage, and it is compared with the normal saline matched group, and there were significant differences (P＜0.01, P＜0.05).Illustrate that this pharmaceutical composition suppository has significantly local hemostasis effect.

The bacteriostatic experiment of 4 pharmaceutical composition suppositorys of experimental example

Adopt the agar doubling dilution to measure the minimum inhibitory concentration (MIC) of this pharmaceutical composition suppository to strain subject.Accurately take by weighing above-mentioned two kinds of preparations by original pharmaceutical content, add to the agar culture medium (being melted up to 50 ℃) that 40ml melts, mixing, therefrom sucking-off 20ml is to the plate of sterilizing, be added into 20ml again in the remaining 20ml agar and do not have medicine agar mixing, again therefrom sucking-off 20ml to the next plate, by that analogy, preparation pastille final concentration is 1000,500,250,125 ... 0.03mg the serial plate of crude drug in whole/ml, inoculate instrument (Denley A400) in each plate surface seeding antibacterial with multiple spot respectively again after the cooling, bacterium liquid final concentration is 105CFU/ml.Insert in 37 ℃ and cultivated 18-24 hours, observed result is the minimum inhibitory concentration (MIC) of this bacterium with the least concentration of medicine in the no bacterial growth plate.The final concentration of levofloxacin is 128,64,32 ... 0.03 μ g/ml.

This pharmaceutical composition of table 8 suppository in-vitro antibacterial spectrum (MIC)

As shown in Table 8: this pharmaceutical composition suppository all has antibacterial activity to staphylococcus aureus, staphylococcus epidermidis, streptococcus pneumoniae, hemophilus influenza, aerobacteria, bacillus pyocyaneus, escherichia coli, Klebsiella Pneumoniae, Bacillus proteus.MIC value scope is 0.125-125mg crude drug/ml.

1. this pharmaceutical composition suppository is better to the antibacterial activity of staphylococcus aureus, staphylococcus epidermidis, hemophilus influenza, aerobacteria, bacillus pyocyaneus, escherichia coli, Klebsiella Pneumoniae, and MIC value scope is 0.25-31.2mg crude drug/ml.

2. this pharmaceutical composition suppository is to Bacillus proteus antibacterial action the best, and MIC value scope is 0.125-1mg crude drug/ml.

Experimental example 5 assays

1. the selection of mobile phase and system suitability experiment

Chromatographic column: Diamonsil C ₁₈(5 μ m, 250 * 0.46mm)

Mobile phase: methanol: acetonitrile: water (50:20:30) (including the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L); Flow velocity: 1ml/min; Column temperature: 30 ℃; Detect wavelength: 346nm.

The result shows, under this chromatographic condition, and in the chromatographic peak, berberine hydrochloride peak shape symmetry, but berberine hydrochloride and other component baseline separation, separating degree is greater than 1.5; Theoretical cam curve (N) is pressed the berberine hydrochloride peak and is calculated, and is more than 4000.

2. measure the selection of wavelength

Berberine hydrochloride reference substance methanol solution is carried out length scanning at 200-400nm, and found that at the 346nm place has absorption maximum, so select this wavelength as measuring wavelength.

3. the investigation of need testing solution preparation method

1. extract choice of Solvent: select for use methanol-hydrochloric acid (100:1) as extracting solvent.

2. the selection of extracting method: compared two kinds of methods of reflux, extract, and supersound extraction in experiment, method is as follows:

Reflux extraction: get this pharmaceutical composition suppository 1.1g, the accurate title, decide, and puts in the round-bottomed flask, accurate methanol-hydrochloric acid (100:1) mixed solution 150ml that adds weighs, and warm macerating is 30 minutes in hot bath, reflux, extract, 2 hours is cooled to room temperature, claims to decide weight again, add methanol-hydrochloric acid (100:1) mixed solution and supply the weight that subtracts mistake, shake up, in-5 ℃ of cold preservations 4 hours, take out the back and filter rapidly, filtrate is put to room temperature, and reuse microporous filter membrane (0.45 μ m) filters, get subsequent filtrate, standby.

Ultrasonic extraction: get this pharmaceutical composition suppository 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, add methanol-hydrochloric acid (100:1) mixed solution 130ml, warm macerating is 30 minutes in hot bath, supersound process (frequency: 40KHz, power: 400W) 60 minutes, be cooled to room temperature, add methanol-hydrochloric acid (100:1) mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse microporous filter membrane (0.45 μ m) filters, and gets subsequent filtrate, and is standby.

Table 9 extraction time is to the influence of berberine hydrochloride

Experimental result shows that supersound extraction efficient is than reflux, extract, height, so this method is selected ultrasonic extraction for use.

3. the selection of extraction time

Get three parts of this pharmaceutical composition suppositorys, every part of 1.1g, the accurate title, decide, put respectively in the 150ml measuring bottle, add methanol-hydrochloric acid (100:1) mixed solution 130ml, warm macerating is 30 minutes in hot bath, respectively supersound process (frequency: 40KHz, power: 400W) 30,60,90 minutes, be cooled to room temperature, add methanol-hydrochloric acid (100:1) mixed solution and be diluted to scale, shake up, in-5 ℃ of cold preservations 4 hours, take out the back and filter rapidly, filtrate is put to room temperature, and reuse microporous filter membrane (0.45 μ m) filters, get subsequent filtrate, standby.Measure by the chromatographic condition of drafting, measurement result sees Table 10.

Table 10 extraction time is to the influence of berberine hydrochloride

The result shows: supersound extraction can be extracted fully in 60 minutes substantially.

In sum, the test sample preparation method is: get 10 of pharmaceutical composition suppositorys of the present invention under the weight differential item, porphyrize is got 1.1g, and accurate the title decides, put in the 150ml measuring bottle, add methanol-hydrochloric acid (100:1) mixed solution 130ml, warm macerating is 30 minutes in hot bath, supersound process (frequency: 40KHz, power: 400W) 1 hour, be cooled to room temperature, add methanol-hydrochloric acid (100:1) mixed solution and be diluted to scale, shake up, in-5 ℃ of cold preservations 4 hours, take out the back and filter rapidly, filtrate is put to room temperature, and reuse microporous filter membrane (0.45 μ m) filters, get subsequent filtrate, promptly.

4. the interference of negative solution experiment

Under selected chromatographic condition, get berberine hydrochloride reference substance solution, need testing solution simultaneously, lack the negative solution sample introduction of Rhizoma Coptidis, found that, in the test sample chromatogram, on the position of identical retention time with the reference substance peak, identical peak is arranged, and negative solution there is not this peak appearance.

5. the investigation of the range of linearity

Accurate respectively absorption concentration is berberine hydrochloride reference substance solution 2 μ l, 5 μ l, 10 μ l, 15 μ l, the 20 μ l of 0.126mg/ml, injects chromatograph of liquid, measures its peak area, and measurement result sees Table 11.

Table 11 reference substance range of linearity measurement result

With peak area value (A) sample size (C) is returned, must the standard curve equation be: y=2939429.69X-25919.46; R=0.9998.To sample size (C) mapping, get a straight line with peak area value (A), the result shows that berberine hydrochloride sample size and peak area linear relationship are good in 0.252 μ g-2.520 μ g scope.

6. precision experiment

Accurate berberine hydrochloride reference substance liquid (0.126mg/ml) the 10 μ l that draw inject chromatograph of liquid, and continuous sample introduction 5 times records the relative standard deviation of berberine hydrochloride peak area, the results are shown in Table 12.

Table 12 precision experimental result

The precision experiment shows: RSD is 0.14%, and this instrument precision is good.

7. stability experiment

The reference substance solution study on the stability: get berberine hydrochloride reference substance liquid (0.126mg/ml) and inject chromatograph of liquid respectively at 0,6,12,24,48 hour accurate 10 μ l of absorption, the record peak area, measurement result sees Table 13.

Need testing solution study on the stability: get pharmaceutical composition of the present invention, prepare need testing solution according to drafting preparation method, inject chromatograph of liquid respectively at 0 hour, 4 hours, 8 hours, 12 hours, the 24 hours accurate 20 μ l of absorption, the record peak area, measurement result sees Table 14.

Table 13 reference substance solution stability experiment result

Table 14 need testing solution stability experiment result

Stability experiment shows that reference substance solution is stable in 48 hours, and need testing solution is stable in 24 hours.

8. repeatability experiment

Get pharmaceutical composition of the present invention, by 5 parts of need testing solutions of preparation method preparation of drafting, the accurate 20 μ l of absorption inject chromatograph of liquid, and the record peak area calculates content of berberine hydrochloride, and measurement result sees Table 15.

The experiment of table 15 repeatability

Repeated experiment shows: adopt this method to extract and detect, repeatability is good.

9. average recovery experiment

Get 6 parts of pharmaceutical composition suppositorys of the present invention, every part of 0.55g, the accurate title, decide, accurate respectively berberine hydrochloride reference substance solution (2.208mg/ml) 1.6,1.6,2.0,2.0,2.4, the 2.4ml of adding, remaining operation prepares application of sample by the test sample preparation method of drafting and reclaims need testing solution, the accurate absorption in the 20 μ l injecting chromatographs measured peak area, the results are shown in Table 16.

The experiment of table 16 average recovery

Response rate experimental result shows: the average average recovery of six experiments is 99.55%, illustrates that the preparation of need testing solution and assay method are reasonable, feasible.

The screening of experimental example 6 semi-synthetic fatty acid glyceride ratios:

Designing this pharmaceutical composition suppository specification is 2.0g, is shaped as bullet shaped; Semi-synthetic fatty acid glyceride and drug powder ratio are made as eight gradients, and effect is selected by experiment, the results are shown in Table 17.

Table 17 substrate and micropowder ratio are selected experiment

As can be drawn from Table 17, select semi-synthetic fatty acid glyceride: drug powder=1.8:1, ratio is suitable, the suppository forming, and hardness is suitable.

Following embodiment all can realize the described effect of above-mentioned experimental example.

The specific embodiment:

Embodiment 1: the preparation of ointment

Rhizoma Coptidis 212g Radix Ampelopsis 231.6g Galla Chinensis 565.5g Radix Curcumae 463g Olibanum 53g.Get above-mentioned five tastes crude drug, add conventional adjuvant, be prepared into ointment according to common process.

Embodiment 2: the preparation of effervescent tablet

Rhizoma Coptidis 160g Radix Ampelopsis 170g Galla Chinensis 360g Radix Curcumae 540g Olibanum 60g Borneolum Syntheticum 27g.Get above-mentioned Six-element crude drug, add conventional adjuvant, be prepared into effervescent tablet according to common process.

Embodiment 3: the preparation of fumigation and washing agent

Rhizoma Coptidis 270g Radix Ampelopsis 260g Galla Chinensis 690g Radix Curcumae 310g Olibanum 45g Borneolum Syntheticum 16g.Get above-mentioned Six-element crude drug, add conventional adjuvant, be prepared into fumigation and washing agent according to common process.

Embodiment 4: the preparation of ointment

Rhizoma Coptidis 212g Radix Ampelopsis 231.6g Galla Chinensis 565.5g Radix Curcumae 463g Olibanum 53g Borneolum Syntheticum 21g.Get above-mentioned Six-element crude drug, add conventional adjuvant, be prepared into ointment according to common process.

Embodiment 5: the preparation of effervescent tablet

Rhizoma Coptidis 270g Radix Ampelopsis 260g Galla Chinensis 690g Radix Curcumae 310g Olibanum 45g.Get above-mentioned five tastes crude drug, add conventional adjuvant, be prepared into effervescent tablet according to common process.

Embodiment 6: the preparation of fumigation and washing agent

Rhizoma Coptidis 160g Radix Ampelopsis 170g Galla Chinensis 360g Radix Curcumae 540g Olibanum 60g.Get above-mentioned five tastes crude drug, add conventional adjuvant, be prepared into fumigation and washing agent according to common process.

Embodiment 7: the preparation of suppository

Rhizoma Coptidis 212g Radix Ampelopsis 231.6g Galla Chinensis 565.5g

Radix Curcumae 463g Olibanum 53g Borneolum Syntheticum 21g

Semi-synthetic fatty acid ester 1286g.

Get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; The decocting that Radix Curcumae, Radix Ampelopsis add 6 times of amounts boils 3 times, the each decoction 1.5 hours, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.10 clear paste that medicinal liquid is concentrated into 60 ℃ of relative densities; Galla Chinensis adds 7 times of water gagings and decocts 3 times, decocts 0.5 hour at every turn, and collecting decoction filters, it is 1.10 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste, is 155 ℃ in inlet temperature, spray drying under 85 ℃ of conditions of leaving air temp is mixed with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester, place evaporating dish, water-bath is dissolved for 40 ℃, adds above-mentioned fine powder, adds the Borneolum Syntheticum mix homogeneously again, pour in the bolt mould, and cooling, the demoulding, suppository is made in packing, every 2g.The consumption usage is: external, one time 1,2 times on the one.

Embodiment 8: the preparation of suppository

Rhizoma Coptidis 160g Radix Ampelopsis 170g Galla Chinensis 360g

Radix Curcumae 540g Olibanum 60g Borneolum Syntheticum 27g

Semi-synthetic fatty acid ester 1213g.

Get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; The decocting that Radix Curcumae, Radix Ampelopsis add 5 times of amounts boils 4 times, the each decoction 1 hour, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.09 clear paste that medicinal liquid is concentrated into to 60 ℃ of relative densities; Galla Chinensis adds 8 times of water gagings and decocts 2 times, decocts 0.5 hour at every turn, and collecting decoction filters, and it is 1.11 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste; In inlet temperature is 151 ℃, spray drying under 82 ℃ of conditions of leaving air temp, and powder mixes with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester and place evaporating dish, water-bath is dissolved for 39 ℃, adds above-mentioned fine powder, adds the Borneolum Syntheticum fine powder again, and mix homogeneously pours in the bolt mould, cooling, and the demoulding, suppository is made in packing, every 2g.The consumption usage is: external, one time 1,2 times on the one.

Embodiment 9: the preparation of suppository

Rhizoma Coptidis 270g Radix Curcumae 310g Galla Chinensis 690g

Olibanum 45g Radix Ampelopsis 260g Borneolum Syntheticum 16g

Semi-synthetic fatty acid ester 1428g.

Get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; The decocting that Radix Curcumae, Radix Ampelopsis add 7 times of amounts boils 2 times, the each decoction 2 hours, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.11 clear paste that medicinal liquid is concentrated into to 60 ℃ of relative densities; Galla Chinensis adds 6 times of water gagings and decocts 4 times, decocts 0.5 hour at every turn, and collecting decoction filters, and it is 1.09 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste; In inlet temperature is 159 ℃, spray drying under 88 ℃ of conditions of leaving air temp, and powder mixes with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester and place evaporating dish, water-bath is dissolved for 41 ℃, adds above-mentioned fine powder, adds the Borneolum Syntheticum fine powder again, and mix homogeneously pours in the bolt mould, cooling, the demoulding, packing, every 2g.The consumption usage is: external, one time 1,2 times on the one.

Embodiment 10: the method for quality control of suppository

Discrimination method:

A, get 1 of embodiment 4 suppository, grind, add 60-90 ℃ of petroleum ether 20ml, supersound process 5 minutes filters, and medicinal residues are waved most solvent, add methanol 20ml, and supersound process 15 minutes filters, and filtrate is as need testing solution.Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=7:1:2 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get 2 of embodiment 4 suppositorys, add water 40ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1-2, and with ether extraction 2 times, each 20ml merges ether solution, volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=6:4:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Embodiment 11: the method for quality control of suppository

The pharmaceutical composition of getting embodiment 5 carries out assay.

Chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=50:20:30 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000;

The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get 10 of this pharmaceutical composition suppositorys under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=100:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=100:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 10-20 μ l of drawing, inject chromatograph of liquid, measure, promptly; Every of pharmaceutical composition suppository of the present invention contains Rhizoma Coptidis with berberine hydrochloride C ₂₀H ₁₈ClNO ₄Meter must not be less than 12mg.

Embodiment 12: the method for quality control of suppository

Discrimination method:

A, get 1 of embodiment 6 suppository, grind, add 60-90 ℃ of petroleum ether 20ml, supersound process 5 minutes filters, and medicinal residues are waved most solvent, add methanol 20ml, and supersound process 15 minutes filters, and filtrate is as need testing solution.Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=7:1:2 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get 2 of embodiment 6 thing suppositorys, add water 40ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1-2, and with ether extraction 2 times, each 20ml merges ether solution, volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=6:4:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.

Assay:

Chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=42:23:27 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000; The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get 10 of this pharmaceutical composition suppositorys under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=80:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=120:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 10-20 μ l of drawing, inject chromatograph of liquid, measure, promptly; It is C with the chemical equation that every of pharmaceutical composition suppository of the present invention contains Rhizoma Coptidis ₂₀H ₁₈ClNO ₄The berberine hydrochloride meter, must not be less than 12mg.

Embodiment 13: the method for quality control of suppository

Discrimination method:

A, get embodiment 4 thing suppository 2g, grind, add 60-90 ℃ of petroleum ether 17ml, supersound process 6 minutes filters, and medicinal residues are waved most solvent, add methanol 23ml, and supersound process 12 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 2.6ml, and supersound process 18 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.4mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=4:1:3 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

Assay:

Chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=48:16:29 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the berberine hydrochloride reference substance, adds methanol and make the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get 10 of this pharmaceutical composition suppositorys under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=122:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=80:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 10-20 μ l of drawing, inject chromatograph of liquid, measure, promptly; It is C with the chemical equation that every of pharmaceutical composition suppository of the present invention contains Rhizoma Coptidis ₂₀H ₁₈ClNO ₄The berberine hydrochloride meter, must not be less than 12mg.

Claims

1, a kind of pharmaceutical composition for the treatment of hemorrhoid is characterized in that the crude drug of this pharmaceutical composition consists of:

Rhizoma Coptidis 150-280 weight portion Radix Ampelopsis 150-280 weight portion Galla Chinensis 350-700 weight portion

Radix Curcumae 300-550 weight portion Olibanum 40-65 weight portion.

2, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug of this pharmaceutical composition consists of:

Rhizoma Coptidis 212 weight portion Radix Ampelopsis 231.6 weight portion Galla Chinensiss 565.5 weight portions

Radix Curcumae 463 weight portion Olibanums 53 weight portions.

3, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug of this pharmaceutical composition consists of:

Rhizoma Coptidis 160 weight portion Radix Ampelopsis 170 weight portion Galla Chinensiss 360 weight portions

Radix Curcumae 540 weight portion Olibanums 60 weight portions.

4, pharmaceutical composition as claimed in claim 1 is characterized in that the crude drug of this pharmaceutical composition consists of:

Rhizoma Coptidis 270 weight portion Radix Ampelopsis 260 weight portion Galla Chinensiss 690 weight portions

Radix Curcumae 310 weight portion Olibanums 45 weight portions.

5, a kind of pharmaceutical composition for the treatment of hemorrhoid is characterized in that the crude drug of this pharmaceutical composition consists of:

Radix Curcumae 300-550 weight portion Olibanum 40-65 weight portion Borneolum Syntheticum 15-30 weight portion.

6, pharmaceutical composition as claimed in claim 5 is characterized in that the crude drug of this pharmaceutical composition consists of:

Radix Curcumae 463 weight portion Olibanums 53 weight portion Borneolum Syntheticums 21 weight portions.

7, pharmaceutical composition as claimed in claim 5 is characterized in that the crude drug of this pharmaceutical composition consists of:

Radix Curcumae 540 weight portion Olibanums 60 weight portion Borneolum Syntheticums 27 weight portions.

8, pharmaceutical composition as claimed in claim 5 is characterized in that the crude drug of this pharmaceutical composition consists of:

Radix Curcumae 310 weight portion Olibanums 45 weight portion Borneolum Syntheticums 16 weight portions.

9, a kind of pharmaceutical composition for the treatment of hemorrhoid is characterized in that adding the semi-synthetic fatty acid ester of 1200-1500 weight portion and makes suppository in following crude drug:

10, pharmaceutical composition as claimed in claim 9 is characterized in that adding the semi-synthetic fatty acid ester of 1286 weight portions and makes suppository in following crude drug:

11, pharmaceutical composition as claimed in claim 9 is characterized in that adding the semi-synthetic fatty acid ester of 1213 weight portions and makes suppository in following crude drug:

12, pharmaceutical composition as claimed in claim 9 is characterized in that adding the semi-synthetic fatty acid ester of 1428 weight portions and makes suppository in following crude drug:

13,, it is characterized in that this pharmaceutical composition adds conventional adjuvant, is prepared into external suppository, spray, ointment, fumigation and washing agent or effervescent tablet according to common process as the arbitrary described pharmaceutical composition of claim 1-8.

14, preparation of drug combination method as claimed in claim 13 is characterized in that this method is: get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; Radix Curcumae, Radix Ampelopsis add the decocting that 4-8 doubly measures and boil 2-4 time, the each decoction 1-2 hours, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.08-1.12 clear paste that medicinal liquid is concentrated into 60 ℃ of relative densities; Galla Chinensis adds 5-9 times of water gagings and decocts 2-4 time, decocted 0.5-1 hour at every turn, and collecting decoction, filtration, it is 1.08-1.12 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste; Be 150-160 ℃ in inlet temperature, spray drying under 80-90 ℃ of conditions of leaving air temp is mixed with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester, place evaporating dish, 40 ± 2 ℃ of fusings of water-bath add above-mentioned fine powder, add Borneolum Syntheticum again, and mix homogeneously pours in the bolt mould, cooling, and suppository is made in the demoulding.

15, preparation of drug combination method as claimed in claim 14 is characterized in that this method is: get Rhizoma Coptidis, frankincense powder is broken into fine powder, is ground into micropowder again, Borneolum Syntheticum is ground into fine powder, and is standby; The decocting that Radix Curcumae, Radix Ampelopsis add 6 times of amounts boils 3 times, the each decoction 1.5 hours, filter, merge above-mentioned filtrate, being concentrated into 60 ℃ of relative densities is 1.20, put cold, adding ethanol makes it to contain the alcohol amount becomes 60%, leaves standstill 14 hours, filters, reclaim ethanol, it is 1.10 clear paste that medicinal liquid is concentrated into 60 ℃ of relative densities; Galla Chinensis adds 7 times of water gagings and decocts 3 times, decocts 0.5 hour at every turn, and collecting decoction filters, it is 1.10 clear paste that filtrate is concentrated into 20 ℃ of relative densities, merges above-mentioned clear paste, is 155 ℃ in inlet temperature, spray drying under 85 ℃ of conditions of leaving air temp is mixed with Rhizoma Coptidis, Olibanum micropowder, is ground into fine powder; Get semi-synthetic fatty acid ester, place evaporating dish, water-bath is dissolved for 40 ℃, adds above-mentioned fine powder, adds the Borneolum Syntheticum mix homogeneously again, pour in the bolt mould, and cooling, the demoulding, suppository is made in packing.

16, the method for quality control of pharmaceutical composition suppository as claimed in claim 13 is characterized in that this method comprises one or more in following discriminating and/or the assay:

Differentiate: A, get this pharmaceutical composition suppository 2g, grind, add 60-90 ℃ of petroleum ether 15-25ml, supersound process 4-6 minutes filters, and medicinal residues are waved most solvent, add methanol 15-25ml, and supersound process 10-20 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 2.5-7.5ml, and supersound process 10-20 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.3-0.7mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=4-10: be developing solvent at 1: 1-3, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get this pharmaceutical composition suppository 4g, add water 30-50ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1-2, uses ether extraction 1-3 times, each 15-25ml, merge ether solution, volatilize, residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=4-8:3-5:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

Assay: chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=40-50:15-25:25-30 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000; The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get this pharmaceutical composition suppository 20g under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=75-125:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=75-125:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and need testing solution 10-20 μ l of drawing, inject chromatograph of liquid, measure, promptly; The every gram of this pharmaceutical composition suppository contains Rhizoma Coptidis with berberine hydrochloride C ₂₀H ₁₈ClNO ₄Meter must not be less than 6mg.

17, the method for quality control of pharmaceutical composition suppository as claimed in claim 16 is characterized in that method comprises one or more in following discriminating and/or the assay:

Differentiate: A, get this pharmaceutical composition suppository 2g, grind, add 75 ℃ of petroleum ether 20ml, supersound process 5 minutes filters, and medicinal residues are waved most solvent, add methanol 20ml, and supersound process 15 minutes filters, and filtrate is as need testing solution; Other gets Rhizoma Coptidis control medicinal material 50mg, adds methanol 5ml, and supersound process 15 minutes filters, and filtrate is medical material solution in contrast; Get the berberine hydrochloride reference substance again, add methanol and make the solution that every 1ml contains 0.5mg, in contrast product solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned three kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with n-butyl alcohol-glacial acetic acid-water=7:1:2 is developing solvent, launches, and takes out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical yellow fluorescence speckle; With the corresponding position of reference substance chromatograph on, show an identical yellow fluorescence speckle;

B, get this pharmaceutical composition suppository 4g, add water 40ml, warmly make dissolving, cool off in the ice bath, filter, filtrate adds hydrochloric acid adjust pH to 1.5, and with ether extraction 2 times, each 20ml merges ether solution, volatilizes, and residue adds methanol 1ml makes dissolving, as need testing solution; Other gets the gallic acid reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; According to thin layer chromatography test, draw each 4 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the sodium carboxymethyl cellulose, with chloroform-ethyl acetate-formic acid=6:4:1 is developing solvent, launches, and takes out, dry, spray develops the color with 2% ferric chloride alcoholic solution; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;

The assay chromatographic condition is tested with system suitability: with the octadecyl bonding mutually silica gel be filler; Methanol: acetonitrile: water=50:20:30 is a mobile phase, wherein includes the sodium lauryl sulphate of 0.05mol/L, the tartaric acid of 0.037mol/L; The detection wavelength is 346nm; Number of theoretical plate calculates by the berberine hydrochloride peak should be not less than 4000; The preparation of reference substance solution: precision takes by weighing the berberine hydrochloride reference substance, adds methanol and makes the solution that contains 0.10mg among every 1ml, promptly; The preparation of need testing solution: get this pharmaceutical composition suppository 20g under the weight differential item, porphyrize, get 1.1g, the accurate title, decide, and puts in the 150ml measuring bottle, adds the mixed solution 130ml of methanol-hydrochloric acid=100:1, warm macerating is 30 minutes in hot bath, through frequency: 40KHz, power: 400W supersound process 1 hour is cooled to room temperature, add methanol-hydrochloric acid=100:1 mixed solution and be diluted to scale, shake up,, take out the back and filter rapidly in-5 ℃ of cold preservations 4 hours, filtrate is put to room temperature, reuse 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, promptly; Algoscopy: accurate respectively reference substance solution 10 μ l and the need testing solution 15 μ l of drawing, inject chromatograph of liquid, measure, promptly; The every gram of this pharmaceutical composition suppository contains Rhizoma Coptidis with berberine hydrochloride C ₂₀H ₁₈ClNO ₄Meter must not be less than 6mg.

18, as the application of the arbitrary described pharmaceutical composition of claim 1-12 in preparation treatment hemorrhoid or anal operation of postop. local swelling, pain, hemorrhage medicine.

19, the application of pharmaceutical composition as claimed in claim 13 in preparation treatment hemorrhoid or anal operation of postop. local swelling, pain, hemorrhage medicine.