CN102973693A

CN102973693A - Pharmaceutical composition used for relaxing bowels to lose weight, and preparation method and application thereof

Info

Publication number: CN102973693A
Application number: CN201210413227XA
Authority: CN
Inventors: 胡昌江; 李倩
Original assignee: Chengdu University of Traditional Chinese Medicine
Current assignee: Chengdu University of Traditional Chinese Medicine
Priority date: 2011-10-27
Filing date: 2012-10-25
Publication date: 2013-03-20
Anticipated expiration: 2032-10-25
Also published as: CN102973693B

Abstract

The invention provides a pharmaceutical composition used for relaxing bowels to lose weight. The pharmaceutical composition is an external medication prepared by using raw rhubarb, cortex magnoliae officinalis, fructus aurantii immaturus and mirabilite as raw material drugs. The invention also provides a preparation method and an application of the pharmaceutical composition. The pharmaceutical patch is researched and developed from decades of clinical experience, on the basis of a theory that six bowels take unobstruction as their normal functional state in traditional Chinese medicine. Stubborn stools are removed through dual functions of the drugs and acupuncture points; the pharmaceutical composition increases intestinal peristalsis, thereby facilitating fat transformation on intestinal walls; the relaxed abdominal muscles are tightened; and elasticity of the abdominal muscles is enhanced, so that the object of ideal rehabilitation and health care effects of relaxing the bowels and losing weight can be achieved. The pharmaceutical patch prevents side effects of stomachache and the like caused by taking purgating drugs such as rhubarb and the like orally. The pharmaceutical patch is convenient for use; and a person can stop taking the patch at any time and stop taking the patch when the person is recovered from the disease.

Description

A kind of pharmaceutical composition for relieving constipation fat-reducing and its production and use

Technical field

The present invention relates to a kind of pharmaceutical composition for the relieving constipation fat-reducing, belong to drug world.

Background technology

According to investigations, nearly 28～30% people is perplexed by the different constipation of degree in the world, is no lack of the weak people such as child, old man among this part crowd, and the crowd of constipation obesity is more and more.Present domestic Overweight and obesity crowd quantity is respectively 200,000,000 and 9,000 ten thousand, and the obesity that causes owing to constipation also sharply increases, it is the main path that vivotoxin and waste product are got rid of that stool drains, if prolonged constipation also can cause a series of diseases such as hemorrhoid even intestinal cancer.Treatment for constipation, it is the oral formulations take Radix Et Rhizoma Rhei as monarch drug in the market, because Radix Et Rhizoma Rhei rushes down lower violent, more people takes side effect such as can not bearing stomachache, and external preparation can be avoided above-mentioned situation, and the external use plaster exploitation in conjunction with the Traditional Chinese Medicine acupoint therapy is gone on the market seldom on the market.

The umbilicus therapy is that China is traditional, one of the Therapeutic Method that shows unique characteristics, and it has long history and abundant content.Existing umbilicus is filled out the records such as medicine, coating in the medical literature of China's earliest extant---the book copied on silk " 52 Bingfang ".Describe warm umbilicus in Han dynasty doctor's family's Zhang Zhongjing " Medical Treasures of the Golden Chamber " and treat sick method.The Jin Daige flood has umbilicus to treat the record of cholera every the real moxibustion treatment of salt in handbook of Prescription for Emergency.The Chinese medicine meridian theory is set forth the Shenque Qizhong of living in caves, and belongs to conception vessel, appoints, superintends and directs, rushes three arteries and veins and have " source three forks " relation, so umbilicus and three pulse classic gas phases are led to.Be through between the twelve regular channels owing to traverse during eight extra-channel is vertical, contact whole body passages through which vital energy circulates, therefore Shenque is spreaded manuer in holes and control functions such as to affect the vital organs of the human body, extremity bones of the body collectively, face nine orifices, skin and flesh muscles and bones, and reach the effect of orderliness negative and positive, circulation of qi promoting and blood, dispeling disease and evil, rehabilitation body.The modern medicine study prompting: umbilicus is last closed place, abdominal cavity in embryo development procedure, compare without subcutaneous fat with whole skin, horny layer of epidermis is thin, barrier function is the most weak, medicine is easy to penetrate disperse, up and down vein of abdominal cavity is arranged under the umbilicus, and abundant vasoganglion is arranged, sensitivity to medicine is the strongest, absorbs rapidly.

Transdermal drug delivery system (Transdermal Drug Delievry Sys tems, TDDS) be a kind of new drug-supplying system that developed recently gets up, it makes medicine absorb by capillary of skin with certain speed and enters the class preparation that the body circulation produces drug effect, it is a kind of dosage form that controlled release drug passes through intact skin, utilize skin as the door of input body circulation, the therapeutical effect of performance medicine is the important breakthrough to traditional dermatologic idea.TDS has avoided liver first-pass effect, medicine gastrointestinal destruction etc.

Application number: 200510021359.8, denomination of invention: the pure Chinese traditional medicine navel of relieving constipation lowering blood-fat and reducing weight pastes; The invention discloses a kind of external application Chinese medicine note agent of the diseases such as cave employing stickup treatment constipation, obesity, hyperlipidemia of namely fainting umbilicus god, its component is Poria, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Fructus Cannabis, Radix Et Rhizoma Rhei, Folium Nelumbinis, Fructus Crataegi, Semen Cassiae; The present invention has the effect of circulation of qi promoting relieving constipation, the dry temperature of heat clearing away, lowering blood-fat and reducing weight, one medicine multiple-effect, compound recipe is effective, and the dual function that takes full advantage of medicine and acupuncture point is evident in efficacy to diseases such as constipation, obesity, hyperlipidemia, external has no side effect, decrease just worry without impairment of YIN, and is easy to use.Application number: 200810071234.X, denomination of invention: Umbilicus plastering powder for defaecation, the present invention relates to contain the medicinal preparation that derives from the vegetable material product, especially belong to a kind of Umbilicus plastering powder for defaecation for the treatment of the constipation disease.The technical solution used in the present invention is: it is that one-tenth by following weight ratio is distributed into medicament: Radix Pseudostellariae, Radix Ophiopogonis, Radix Rehmanniae, Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Fructus Cannabis, Semen Armeniacae Amarum, Fructus Gleditsia, Herba Cistanches, Radix Angelicae Sinensis, Matrii Sulfas Exsiccatus, Semen Arecae, Natrii Sulfas, Folium Sennae.Beneficial effect of the present invention is: the theoretical basis that the present invention is based on traditional medicine " inner disease outer treat ", with the administration of umbilicus cave, avoided the stimulation to liver, kidney, stomach, not only solved the oral medication pathogenic QI in the body and property of medicine phase cell is refused, medicine enters stomach namely to be told, and can not receive the drawbacks such as medicine.Avoid simultaneously the internal organs official to the decomposition of endo-medicine, also effectively overcome the problem that the patient is difficult for taking medicine, and without any side effects, be worth of widely use.

Summary of the invention

Technical scheme of the present invention has provided a kind of pharmaceutical composition for relieving constipation fat-reducing and its production and use.

The invention provides a kind of pharmaceutical composition for relieving constipation fat-reducing, it is to be the topical agent that crude drug is prepared from by Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas.

Further preferably, the weight proportion of described crude drug is:

Radix Et Rhizoma Rhei 100-500 part, Cortex Magnoliae Officinalis 50-300 part, Fructus Aurantii Immaturus 50-150 part, Natrii Sulfas 5-40 part.

Further preferably, the weight proportion of described crude drug is:

300 parts of Radix Et Rhizoma Rhei, 150 parts of Cortex Magnoliae Officinalis, 100 parts of Fructus Aurantii Immaturuss, Natrii Sulfas 10-25 part.

It is to be active component by the protogenic medicinal powder of Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas or extractive with organic solvent, adds the medicament that pharmacy external preparation adjuvant commonly used or complementary composition are prepared from.

Wherein, described topical agent is tincture, unguentum, lotion, liniment, cataplasma, patch.

Wherein, comprise in the described patch and sealing up for safekeeping in the medicated powder of breathability sticker and the medicinal liquid of fill; Wherein, the drug loading of medicinal liquid is 1-3ml, and concentration is counted 1-2g/ml with primary crude drug, and the medicated powder drug loading is 1-3g.

Wherein, Sennoside A content is not less than 4.490mg/g in the described medicinal liquid; Be not less than 37.340mg/g with Determination of Magnolol in the medicinal liquid, Determination of Magnolol is not less than 41.830mg/g.

The present invention also provides a kind of method for preparing described pharmaceutical composition, and it comprises the steps:

A, weighting raw materials;

B, take by weighing Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, be ground into coarse powder, added the 65-85% alcohol dipping 6-12 hour, carry out the normal pressure percolation, the percolate natural subsidence is got supernatant and is evaporated to without the alcohol flavor in 30 ℃-40 ℃, be mixed with the medicinal liquid of 1-2g/ml with 20-30% ethanol, the Natrii Sulfas that adds medicinal liquid weight 10～25% makes its dissolving, adds medicinal liquid weight 1-6% azone again, namely gets the medicinal liquid that concentration is primary crude drug meter 1-2g/ml;

C, take by weighing Radix Et Rhizoma Rhei, Natrii Sulfas, be ground into fine powder, fine powder, middle powder, or coarse powder, mix homogeneously, sterilization namely gets medicated powder;

D, medicated powder is sealed up for safekeeping in the breathability sticker, liquid medicine filling is prepared into patch.

Further preferably, it comprises the steps:

A, weighting raw materials;

B, take by weighing respectively Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas in the ratio of 3:1.5:1, be ground into coarse powder, add 75% alcohol dipping 6 hours, carry out the normal pressure percolation, the percolate natural subsidence is got supernatant and is evaporated to without the alcohol flavor in 40 ℃, be mixed with the medicinal liquid of 1g/ml with 25% ethanol, add 10～25% Natrii Sulfas and make its dissolving, add 2% azone, namely get the medicinal liquid that concentration is primary crude drug meter 1g/ml;

C, take by weighing an amount of Radix Et Rhizoma Rhei, Natrii Sulfas in 1: 1 ratio, be ground into fine powder, mix homogeneously, sterilization namely gets medicated powder;

The present invention also provides the purposes of described pharmaceutical composition in the external used medicine of preparation relieving constipation fat-reducing.

It is with a long history that Radix Et Rhizoma Rhei is used as medicine, and " Dragon Lord book on Chinese herbal medicine warp " classifies low-grades as, and meaning " lower blood stasis, blood closes, cold and heat, broken lumps in the chest and abdomen, the prolonged fluid-retention dyspepsia is cleansed the intestines and stomach, pushes away Chen Zhixin, tonneau water paddy, relieving dyspepsia in the accent, peace and the five internal organs." " Bencao Jingshu " also think: " and Radix Et Rhizoma Rhei abnormal smells from the patient large bitter Great Cold, the property official report is directly satisfied, be longer than lower logical, so for rushing down lower two Jiao in typhoid fever epidemic febrile disease, calentura, damp and hot, the accumulation of heat, two just can not, and damp and hot glue expectorant under two Jiao's key medicine ".In this prescription, the Radix Et Rhizoma Rhei bitter cold is logical falls, purging heat to relax the bowels, and it is stagnant to cleanse Gastrointestinal excess heat, to control heat, reality, is monarch drug.Knot in the excess-heat, the internal organs vapour lock stagnates, so with the Cortex Magnoliae Officinalis therapeutic method to keep the adverse QI flowing downwards except full, make the logical row of falling of gastrointestinal mechanism of qi to help relieving constipation by purgation, be adjuvant drug.Medicine material of the present invention rushes down lower lays equal stress on circulation of qi promoting, rushes down the next gastrointestinal mechanism of qi that passes unimpeded, and circulation of qi promoting then helps to rush down time, hereat rush down exert oneself high.

Medicine patch of the present invention is the theory according to motherland's medical science the six hollow viscera must keep its unobstructed, collection many decades clinical experience develops with great concentration, dual function by medicine and acupuncture point is removed stool, increase enterokinesia, be convenient to fat conversion on the intestinal wall, the abdominal muscles of take-up slack, and increase its elasticity, reach desirable relieving constipation, the purpose of fat-reducing rehabilitation.Medicine patch of the present invention has been avoided the side effect such as stomachache of the purgatives medicines such as Radix Et Rhizoma Rhei due to oral, and is easy to use, at any time drug withdrawal, Discontinue medication as soon as get effect.

Description of drawings

Fig. 1 length scanning is figure as a result

Fig. 2 skin of rat figure

Fig. 3 improved Franz diffusing cells method figure

The solubility curve figure of Fig. 4 sodium sulfate

Fig. 5 Sennoside A reference substance HPLC spectrogram

The negative HPLC spectrogram of Fig. 6 reference substance

The negative HPLC spectrogram of Fig. 7 test sample

Fig. 8 need testing solution HPLC spectrogram

Fig. 9 magnolol, honokiol reference substance HPLC spectrogram

The negative HPLC spectrogram of Figure 10 reference substance

The negative HPLC spectrogram of Figure 11 test sample

Figure 12 Cortex Magnoliae Officinalis test sample HPLC spectrogram

The specific embodiment

The preparation of embodiment 1 external use plaster of the present invention

1. medicine patch preparation technology of the present invention

Medicine patch of the present invention is divided into solid drugs and medicinal liquid two parts, and solid drugs is sealed up for safekeeping in the breathability sticker, removes the sticker backing layer during administration, drips medicinal liquid at medicated powder, is affixed on umbilicus and gets final product.

1.1 medical material, instrument and reagent

1.1.1 the medical materials such as medical material Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis all are purchased from Chengdu new Flos Nelumbinis decoction pieces company, meet standards of pharmacopoeia through discriminating and assay ^[1]

1.1.2 instrument Shimadzu LC-20A high performance liquid chromatograph, Sai Duolisi Bp211DAG electronic balance (German Sartorius), UV-1100 ultraviolet-uisible spectrophotometer (Shanghai Techcomp Instrument Ltd.), ultrasonic cleaner CX-100(Beijing armarium two factories), the multiplex vacuum pump of SHD-III type circulating water type (high and new technology industrial development zone, Baoding sunlight Science ﹠ Teaching Instrument factory), rotary evaporator RE-52A(Shanghai Yarong Biochemical Instrument Plant), powder beater (Shanghai is product industry and trade company limited for a long time).

1.1.3 medicine and reagent

Medicine emodin reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110756-200110), Sennoside A reference substance (Chengdu Man Site bio tech ltd, lot number: A0182, purity 〉=98%), magnolol reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 110729-200309), honokiol reference substance (Nat'l Pharmaceutical ﹠ Biological Products Control Institute, lot number: 0730-200206).

The reagent magnesium acetate is produced by the Beijing Chemical Plant, sodium bicarbonate is produced by the Long Huagongshijichang of Chengdu section, and 95% ethanol, glacial acetic acid, methanol, petroleum ether (60～90) etc. are produced by the Long Huagongshijichang of Chengdu section, and methanol is chromatograph methanol, water is double distilled water, and all the other reagent are analytical pure.

1.2 the preparation of solid medicated powder

Get Radix Et Rhizoma Rhei, mirabilite powder is broken into fine powder, in the ratio mix homogeneously of 1:1, and get final product.

1.3 the preparation of medicinal liquid

Medicinal liquid is the extracting solution of Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas.

1.3.1 evaluation index

The anthra-glucoside of bonding state is the effective ingredient under rushing down in the Radix Et Rhizoma Rhei, mainly comprises anthraquinone glycoside and dianthracene ketoside, and is in the majority with the dianthracene ketoside, and Sennoside A, B, C, D, E, F are arranged in the dianthracene ketoside, and wherein Sennoside A content is the highest ^[1]So, with combined anthraquinone, Sennoside A as the technology assessment index.

Stool drains the promotion by " gas ", and the circulation of qi promoting effect of Cortex Magnoliae Officinalis be can not ignore in the side, so choose magnolol, two index one-tenth of honokiol assign to estimate technique.

1.3.2 content assaying method

1.3.2.1 the content of combined anthraquinone in the determined by ultraviolet spectrophotometry Radix Et Rhizoma Rhei ^[2-3]

Combined anthraquinone content=total anthraquinones content-sequestered anthraquinone content.

The preparation of emodin reference substance solution

Get the about 1mg of emodin reference substance, accurately weighed, put in the 10ml volumetric flask, with dissolve with methanol and be settled to scale, shake up, and get final product.

Measure determining of wavelength

The accurate emodin reference substance solution 0.5ml that draws puts in the evaporating dish, fling to methanol, add 1.0% magnesium acetate dissolve with methanol solution residue, and standardize solution is put in the 10ml volumetric flask, shake up, make reference with 1.0% magnesium acetate methanol solution, in 300～900nm wave band interscan emodin reference substance solution, it is good that the result is presented at 504nm place absworption peak, the results are shown in Figure 1.And blank solvent at the 504nm place almost without absorbing, interference measurement result not is so select 504nm as measuring wavelength.

The drafting of standard curve

Emodin reference substance solution 0.2,0.4,0.6,0.8,1.0ml put in the evaporating dish in accurate the absorption respectively, volatilize solvent, add 1.0% magnesium acetate dissolve with methanol solution residue, and standardize solution are put the 10ml volumetric flask.Take methanol as blank solvent, adopt spectrophotometry, measure its absorbance (A) at the 504nm place, the results are shown in Table 1.

The absorbance of table 1 emodin reference substance solution variable concentrations

Take emodin reference substance solution concentration C (mg/ml) as abscissa, take absorbance (A) as vertical coordinate, getting regression equation is Y=40.821X+0.0094, R ²=0.9996, experimental result shows that emodin reference substance content is the good linear relation in 0.022～0.224mg scope.

Assay of Total Anthraquinone

The accurate extracting solution 1ml that draws, standardize solution is put in the 50ml volumetric flask, shakes up, filter, precision is measured subsequent filtrate 5ml, and decompression volatilizes solvent, ice acetic acid-25% hydrochloric acid (5:1) 10ml, reflux 2h, cooling is put in the separatory funnel, uses chloroform extraction 2 times, each 10ml, merge the chloroform layer, and wash with massive laundering, reclaim solvent to doing, residue adds 1.0% magnesium acetate dissolve with methanol solution, standardize solution is put in the 50ml measuring bottle, shakes up, take 1.0% magnesium acetate methanol solution as blank solution, measure its absorbance in the 504nm place, calculate the content of general anthraquinone.

The sequestered anthraquinone content is measured

The above-mentioned subsequent filtrate 5ml of accurate absorption puts in the evaporating dish, volatilizes solvent, and residue adds a small amount of water washing, and shifts and put in the separatory funnel, and all the other operate with associative operation under the Assay of Total Anthraquinone item, measures its absorbance in the 504nm place, calculates the sequestered anthraquinone content.

1.3.2.2HPLC method is measured the content of Sennoside A ^[4-6]

Chromatographic condition

Chromatographic column: Comatex-C ₁₈Post (5 μ m, 250mm * 4.6mm);

Detect wavelength: 340nm;

Mobile phase: methanol: water (50:50);

Flow velocity: 0.5ml/min;

Column temperature: 30 ℃.

The preparation of Sennoside A reference substance solution

It is an amount of to get the Sennoside A reference substance, accurately weighed, adds methanol and is mixed with the reference substance solution that every 1ml contains the 50ug Sennoside A, and get final product.

The preparation of need testing solution

The accurate extracting solution 1ml that draws adds methanol constant volume and puts in the 10ml volumetric flask, and get final product.

Assay

Precision is got reference substance solution, each 10 μ l sample introduction of need testing solution respectively, measures, and calculates Sennoside A content.

1.3.2.3HPLC method is measured magnolol, honokiol content

Chromatographic condition ^[7]

Chromatographic column: Comatex-C ₁₈Post (5 μ m, 250mm * 4.6mm);

Mobile phase: methanol-water (78:22);

Detect wavelength: 294nm;

Flow velocity: 1ml/min;

Column temperature is: 35 ℃.

The preparation of reference substance solution

Get magnolol, the honokiol reference substance is an amount of, and is accurately weighed, add methanol and make the solution that every 1ml contains magnolol 28ug, honokiol 30ug, and get final product.

The preparation of need testing solution

The accurate extracting solution 1ml that draws volatilizes solvent, adds dissolve with methanol and standardize solution and puts in the 25ml volumetric flask, and get final product.

Assay

Accurate reference substance solution, each 10 μ l sample introduction of need testing solution drawn are measured respectively, calculate honokiol, Determination of Magnolol.

1.3.3 extraction process screening

1.3.3.1 the orthogonal test factor level is selected

Learn that according to trial test the principal element that affects this percolation extraction process is: concentration of alcohol, ethanol consumption and dip time, so say capable L by 3 factors, 3 levels ₉(3 ⁴) test.Factor level sees Table 2.

Table 2 percolation factor level

1.3.3.2 the preparation of orthogonal test sample

Take by weighing respectively Radix Et Rhizoma Rhei 150g, Cortex Magnoliae Officinalis 75g, Fructus Aurantii Immaturus 50g extracts by the every listed condition of orthogonal test table, and the gained medicinal liquid is suitably concentrated, is settled at last 500ml, and is for subsequent use.

1.3.3.3 the assay of index components

With reference to associative operation under the 1.3.2 item, measure respectively the content of combined anthraquinone, Sennoside A, honokiol, magnolol, the results are shown in Table 3.

1.3.3.4 result of the test

The orthogonal test assay the results are shown in Table 3.

Table 3 orthogonal test assay result

Adopt the aggregative weighted point system, the content of combined anthraquinone, Sennoside A, honokiol, magnolol all with higher for well, so respectively the peak of correspondence is decided to be 100 minutes, should use the high-load value of formula y=Y/.

According to the purpose of experimentation, each component content weight coefficient is respectively: combined anthraquinone content (Y ₁) be 0.3, Sennoside A content (Y ₂) be 0.4, honokiol content (Y ₃), Determination of Magnolol (Y ₄) be 0.15, with four individual event additions, use formula Y=0.3y ₁+ 0.4y ₂+ 0.15y ₃+ 0.15y ₄, obtain the value of respectively testing comprehensive grading, the results are shown in Table 4.

Table 4 orthogonal test calendar and result

1.3.3.5 data analysis

The results of analysis of variance sees Table 5.

The variance analysis of table 5 comprehensive grading

By the results of analysis of variance as can be known, concentration of alcohol (A) has a significant impact the result, and ethanol consumption (B) and dip time (C) impact are not remarkable, and be easy and simple to handle in order to make, and is fit to large production, so select B ₁, C ₁, the final extraction process of determining is A ₂B ₁C ₁, namely the alcohol dipping of 6 times of amounts 75% was carried out the normal pressure percolation in 6 hours.

1.3.3.6 confirmatory experiment

In order to verify the accuracy of the above results, guarantee the reasonable of extraction process, carry out demonstration test according to above-mentioned definite optimal extract process, the results are shown in Table 6.

Table 6 demonstration test result

Confirmatory experiment is the result show: this extraction process science, reasonable, feasible.

1.3.4 the purification of medicinal liquid

Percolate needs further purification, the feasibility of the reality of considering, this part is intended adopting centrifuging (different centrifugation time) and natural sedimentation to carry out remove impurity and is investigated, and take combined anthraquinone, Sennoside A, honokiol and magnolol as evaluation index, to determine best impurity removal process, experimental result sees Table 7.

Table 7 purification experimental result

Can be found out that by above result centrifuging and natural sedimentation are not remarkable to the loss difference of above-mentioned each index, but consider that from the angle of large production the natural sedimentation cost is minimum and more easy to operate, so the final purifying process of determining is natural subsidence 24h.

1.3.5 the concentrated experiment of medicinal liquid

Percolate is behind natural subsidence 24h, and supernatant adopts the mode of concentrating under reduced pressure to concentrate, because main effective ingredient combined anthraquinone and magnolol etc. are subject to the impact [8] of temperature, so the temperature of concentrating under reduced pressure is investigated, experimental result sees Table 8.

Table 8 concentrating under reduced pressure temperature is investigated the result

Can be found out that by above-mentioned experimental result temperature is all influential to combined anthraquinone, magnolol etc., temperature is higher, loss more.When thickening temperature was 30 ℃, what reach medicinal liquid required the required time of concentration longer, and namely heated time increases, so compare the more of loss with 40 ℃, it is 40 ℃ that historical facts or anecdotes is tested the final thickening temperature of determining.

1.3.6 brief summary and discussion

By above-mentioned experimentation, the final medicinal liquid extraction process of determining is: Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus are ground into coarse powder, add the alcohol dipping 6 hours of 6 times of amounts 75%, the normal pressure percolation, and percolate natural subsidence 24h is in 40 ℃ of concentrating under reduced pressure.

2. moulding process examination

Transdermal patch is by membrane polymer, framework material, the materials such as pressure sensitive adhesive form, be divided into membrane controlled release type, viscose decentralized, matrix diffusion type etc., contain again different substrate in the Drug Storage of all kinds transdermal patch, in present research report, also do not have generally acknowledged moulding process measurement index, the substrate prescription is also different.The medicine patch of the present invention of this experimentation is played a role jointly by solid medicated powder and medicinal liquid two parts, so the grinding particle size that contains alcohol amount, medicated powder of the drug loading of medicated powder and medicinal liquid, the concentration of medicinal liquid, medicinal liquid seems particularly important to the transmitance of medicine, so need further to investigate by the transdermal test of exsomatizing, concrete grammar is as follows:

2.1 medical material, instrument, reagent and animal

The medical materials such as medical material Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis all are purchased from Chengdu new Flos Nelumbinis decoction pieces company, all meet standards of pharmacopoeia through evaluation and assay ^[7]

Instrument Shimadzu high performance liquid chromatograph (LC-20A Chengdu peace perseverance reaches Science and Technology Ltd.), TK-12B type percutaneous dispersion test instrument (the triumphant scientific and technological trade Co., Ltd of the Shanghai iron of fine quality), Sai Duolisi electronics sky (Bp211DAG Germany Sartorius), ultrasonic cleaner (CX-100 Beijing armarium two factories), powder beater (Shanghai is product industry and trade company limited for a long time).

Medicine and reagent

Medicine Sennoside A reference substance (lot number: A0182, purity 〉=98% Chengdu Man Site bio tech ltd).

Reagent azone (chemical pure CP), sodium sulfide, chloral hydrate, normal saline are produced by the Long Huagongshijichang of Chengdu section; Benzalkonium bromide solution is produced by Nanchang white clouds pharmaceutcal corporation, Ltd.

95% ethanol, glacial acetic acid, methanol, petroleum ether (60～90) etc. are produced by the Long Huagongshijichang of Chengdu section.

Methanol is chromatograph methanol, and water is double distilled water, and all the other reagent are analytical pure.

Animal SD rat, ♀ ♂ has concurrently, body weight (150 ± 10) g, Chengdu reaches large bio tech ltd to be provided, the quality certification number: the SCXK(river) 2008-24.

The transdermal test method 2.2 exsomatize

2.2.1 the preparation of isolated skin

Rat is applied to skin of abdomen through lumbar injection 10% chloral hydrate (normal saline preparation) anesthesia with 8% sodium sulfide solution, cleans with distilled water behind the 1min, raises 24h, and live body strips skin of abdomen after the anesthesia.On skin tiling and clean glass plate, horny layer is carefully removed subcutaneous fat and adhesion thing down, cleans with normal saline, through 0.1% benzalkonium bromide solution soaking 0.5h, clean with normal saline again, place culture dish, add a small amount of normal saline, preserve in 4 ℃ of refrigerator sealings, be finished in 7 days ^[9,10](seeing accompanying drawing 2).

2.2.2 transdermal test

Corium Mus is taken out, blot the normal saline on surface with filter paper, room temperature is transferred the usefulness of purchasing, and skin is fixed on the improved Franz diffusing cells method ^[11](seeing accompanying drawing 3), make stratum corneum side upwards, medicated powder, medicinal liquid are added on the Corium Mus, receiving chamber fills it up with dispersive medium, interlayer is with 37 ℃ of water bath heat preservations, magnetic stir bar stirs with the speed of 200r/min, takes out 1ml receiving liquid (adding simultaneously the equivalent normal saline) respectively at 2h, 4h, 6h, 8h, 10h, 12h, and sample introduction is measured the content of Sennoside A.Calculate the accumulation infiltration capacity by following formula again:

Q=Cn?V/A?Cn＝C1+C2+C3+…………+Cj

Q is the accumulation infiltration capacity in the formula, and A is the reception tank area, and V is the reception tank volume, j=2,4,6,8,10,12.In this experiment, V=13.5mL, A=3.142cm ²

2.2.3 assay

The preparation of need testing solution places evaporating dish with the receiving liquid of taking out, and volatilizes in 60 ℃ of water-baths, and residue adds dissolve with methanol, and shifts and put in the 10ml volumetric flask, standardize solution, and mixing, and get final product.

Measure the method for Sennoside A content according to HPLC method under the 1.3.3 item and measure, calculate the content of Sennoside A.

All isolated tests of this section all operate as stated above, and concrete the investigation thes contents are as follows:

2.3 drug loading experiment

Guarantee the curative effect of patch, it is particularly important that drug loading seems, so need the drug loading of this patch is investigated.Transdermal is tested each sample volume seldom owing to exsomatize, be difficult to measure accurately the content of combined anthraquinone and Sennoside A, honokiol, magnolol, so take the representative species Sennoside A of main effective ingredient as evaluation index, carry out the investigation of drug loading with orthogonal test.

Take extract concentration, medicinal liquid drug loading, medicated powder drug loading as the investigation factor, carry out L respectively ₉(3 ⁴) test, each factor level sees Table 9.

Table 9 orthogonal experiment factor level

Orthogonal experiments sees Table 10.

Table 10 orthogonal experiment arrangement result (n=3)

The results of analysis of variance sees Table 11.

Table 11 the results of analysis of variance

Known that by the results of analysis of variance main influence factor is the concentration (A) of extracting solution, medicinal liquid drug loading (B) and medicated powder drug loading (C) impact are not remarkable, consider contact and the actual operability of medicated powder and medicinal liquid, finally are defined as A ₁B ₂C ₁, namely liquor strength is 1g/ml, and the medicinal liquid drug loading is 2ml, and the medicated powder drug loading is 1g.

2.4 the medicinal liquid concentration of alcohol is investigated

Concentration of alcohol is larger on the Transdermal absorption impact of medicine ^[12,13], and the phenomenon such as easily go mouldy in the general medicinal liquid storing process, and an amount of ethanol both had been conducive to Transdermal absorption, can reach again the antiseptical purpose, so need to investigate the concentration of alcohol of this medicinal liquid.

By consulting literatures knows that the anticorrosion concentration of ethanol is more than 20%, but the ethanol medicinal liquid of excessive concentration can cause anaphylaxis or zest to the patient, still concentration of alcohol investigation scope be 20～30%.

The preparation of different concentration ethanol medicinal liquid is prepared with the ethanol of variable concentrations, and be get final product with reference to the preparation method that supplies reagent liquid under the 1.3.3 item.

Take Sennoside A as evaluation index, adopt the transdermal experiment that exsomatizes to investigate, wherein liquor strength is 1g/ml, and the medicinal liquid drug loading is 2ml, and the medicated powder drug loading is 1g, and experimental result sees Table 12.

Table 12 medicinal liquid concentration of alcohol is investigated result (n=3)

By above-mentioned experimental result as can be known, the medicinal liquid osmosis effect that contains 25% ethanol is better, so the concentration of alcohol of medicinal liquid is decided to be 25%.

2.5 the granularity of medicated powder is investigated

This preparation is comprised of medicated powder and medicinal liquid two parts, and medicated powder needs the dissolving competence exertion drug effect of medicinal liquid, so the granularity of medicated powder also is the principal element that affects the Transdermal absorption effect, needs it is investigated.

Take Sennoside A as evaluation index, adopt the transdermal experiment that exsomatizes to investigate, the concentration that wherein contains 25% ethanol medicinal liquid is 1g/ml, and the medicinal liquid drug loading is 2ml, and the medicated powder drug loading is 1g, and experimental result sees Table 13.

Table 13 medicated powder granularity is investigated the result

By above-mentioned experimental result as can be known: when medicated powder was pulverized as fine powder, the Transdermal absorption effect was better, so the medicated powder granularity is decided to be fine powder.

2.6 the dissolution studies of Natrii Sulfas

This product is Sulfates mineral Natrii Sulfas family Natrii Sulfas, the crystalline solid that forms through processing and refining ^[14], direct dissolving mirabilite method is selected in this experiment, Natrii Sulfas is dissolved in the medicinal liquid, so the addition of Natrii Sulfas becomes the factor of the effects.

The solubility curve of sodium sulfate is seen accompanying drawing 4.As seen from the figure, Natrii Sulfas dissolubility and temperature are proportionate, but the dissolubility of Natrii Sulfas in ethanol be much smaller than the dissolubility in water, and this patch herb liquid contains 25% ethanol, so the meltage of Natrii Sulfas is examined or check.

The experiment proved that the dissolubility difference of Natrii Sulfas in extracting solution is little between 10 ℃～30 ℃, its meltage is 10g/100ml～25g/100ml, gets final product so can add 10～25% Natrii Sulfas in medicinal liquid according to practical situation in large the production.

2.7 the concentration of penetration enhancer azone is investigated

Percutaneous absorption enhancer commonly used mainly contains natural penetration enhancer (terpenoid, quintessence oil and lactone etc.); Synthetic penetration enhancer (dimethyl sulfoxide, dimethyl imide, pyrrolidinone compounds, phospholipid and phosphoric acid salt, organic acid and esters, phthalein amine etc.), wherein the most representative be laurocapram (azone, Azone) ^[15-17]Azone not only can promote the Transdermal absorption of most hydrophilic and hydrophobic compound, and is applicable to the Chinese herbs paste of chemical constituent more complicated, so this research selects azone as Percutaneous absorption enhancer, and determines the optimum amount of azone by the transdermal test ^[18]

Take Sennoside A as evaluation index, the transdermal experiment that adopt to exsomatize is investigated, and the concentration that wherein contains 25% ethanol medicinal liquid is 1g/ml, and the medicinal liquid drug loading is 2ml, and medicated powder is fine powder, and the medicated powder drug loading is 1g, and experimental result sees Table 14.

The preparation that contains the azone medicinal liquid is with reference to correlation technique preparation under the 1.4.3 item, is mixed with the medicinal liquid of 1g/ml with 25% ethanol, the adding azone be mixed with respectively 0%, 1%, 2%, 3%, 4%, 5%, 6% contain the azone medicinal liquid.

Table 14 azone concentration is investigated result (n=3)

Take the time as abscissa, Sennoside A accumulation infiltration capacity is the vertical coordinate mapping, gets Sennoside A infiltration equation and penetration curve, the results are shown in Table 15.

Table 15 Sennoside A infiltration equation

Known that by table 15 transdermal effect is better when azone content is 2%, so the content of penetration enhancer azone is decided to be 2% in the medicinal liquid.

2.8 confirmatory experiment

By above-mentioned orthogonal experiment, experiment of single factor has been determined drug loading, medicinal liquid concentration of alcohol, azone content etc., for confirmatory experiment result's correctness, reasonability, carries out confirmatory experiment.

Take Sennoside A as evaluation index, the transdermal experiment that exsomatizes, the concentration that wherein contains 25% ethanol medicinal liquid is 1g/ml, and wherein azone concentration is 2%, and the medicinal liquid drug loading is 2ml; Medicated powder is fine powder, and the medicated powder drug loading is 1g, and experimental result sees Table 16.

Table 16 confirmatory experiment result

The confirmatory experiment result shows above experimental result science, reasonable.

2.9 the preparation of medicated powder

2.9.1 pulverize

Ratio in 1: 1 is got Radix Et Rhizoma Rhei, Natrii Sulfas is an amount of, is ground into fine powder, mix homogeneously.

2.9.2 sterilization treatment

Medicated powder to mix homogeneously carries out the 0.5h ultra-vioket radiation, carries out sterilization treatment.

2.9.3 packing

Medicated powder after the sterilization is packaged in the plaster, whenever mounts 1g.

2.10 brief summary and discussion

The medicine patch preparation technology of the present invention who finally determines by this part experimentation is:

It is an amount of to take by weighing respectively Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus in 3: 1.5: 1 ratios, be ground into coarse powder, the alcohol dipping 6h that adds 6 times of amounts 75%, carry out the normal pressure percolation, percolate natural subsidence 24h gets supernatant and is evaporated to without the alcohol flavor in 40 ℃, be mixed with the medicinal liquid of 1g/ml with 25% ethanol, add 10～25% Natrii Sulfas and make its dissolving, add 2% azone, namely get medicinal liquid.Take by weighing an amount of Radix Et Rhizoma Rhei, Natrii Sulfas in 1: 1 ratio, be ground into fine powder, mix homogeneously, sterilization is sealed up for safekeeping in sticker and is got final product.

The present invention is the brown color patch of uniformity, gas delicate fragrance; Medicinal liquid is sepia, and feeble QI is fragrant.

Function cures mainly: remove stool, be used for the obesity that constipation and constipation cause.

Usage and dosage: paste every day one.Throw off the lid lining, medicinal liquid is dropped on the medicated powder, be affixed on umbilicus.

The assay of embodiment 2 medicine patch of the present invention

1 Sennoside A assay

(1) instrument and reagent

Shimadzu LC-20A high performance liquid chromatograph, ultrasonic cleaner CX-100(Beijing armarium two factories), Sai Duolisi Bp211DAG electronic balance (German Sartorius), the multiplex vacuum pump of SHD-III type circulating water type (high and new technology industrial development zone, Baoding sunlight Science ﹠ Teaching Instrument factory).

Acetonitrile, methanol are chromatographically pure, and water is double distilled water, and all the other reagent are analytical pure.

Sennoside A reference substance (lot number: A0182, purity 〉=98%, Chengdu Man Site bio tech ltd).

(2) chromatographic condition

Chromatographic column: Comatex-C ₁₈Post (5 μ m, 250mm * 4.6mm)

Mobile phase: compared different mobile phases in the trial test, the separating effect of the acetonitrile-water of different proportion, methanol-water (80:20), methanol-water (70:30), methanol-water (60:40), methanol-water (50:50) has selected methanol-water (50:50) to be mobile phase at last.

Flow velocity: 0.5ml/min;

Detect wavelength: 340nm;

Column temperature: 30 ℃.

(3) preparation of reference substance solution

It is an amount of to get the Sennoside A reference substance, accurately weighed, adds methanol and makes every 1ml and contain approximately 50ug Sennoside A reference substance, and get final product.

(4) preparation of need testing solution

The accurate medicinal liquid 0.5ml that draws volatilizes solvent, and residue adds dissolve with methanol and standardize solution is put in the 10ml measuring bottle, and get final product.

(5) preparation of negative sample solution

Get the Radix Et Rhizoma Rhei negative sample, according to the preparation of test sample preparation method, and get final product.

Reference substance, reference substance are negative, test sample is negative and the high-efficient liquid phase chromatogram of test sample is seen accompanying drawing 5～Fig. 8.

(6) linear relationship is investigated

Accurate absorption Sennoside A reference substance solution (0.058mg/ml) respectively sample introduction 0.2,0.5,0.8,1.0,1.2,1.5ul is measured peak area, the results are shown in Table 17.

Table 17 range of linearity is investigated the result

Take Sennoside A reference substance sample size as abscissa, corresponding peak area value is the vertical coordinate curve plotting, and getting Regression Equations is Y=1E+06X-1518.2, R ²=0.9996, show that Sennoside A has good linear relationship between 0.0116～0.0870ug.

(7) precision test

The accurate Sennoside A reference substance solution 10ul that draws repeats sample introduction 6 times, and the record peak area the results are shown in Table 18.

Table 18 Precision test result

Result of the test shows: RSD is 0.13%, less than 2%, so instrument precision is good.

(8) stability test

Get respectively same reference substance solution and same need testing solution, respectively at 0h, 2h, 4h, 8h, 12h sample introduction 10 μ l, the record peak area the results are shown in Table 19,20.

Table 19 reference substance stability test result

Table 20 sample stability result of the test

Result of the test shows: reference substance RSD is 0.16%, and sample RSD is 0.65%, and all less than 2%, so reference substance solution and sample solution are stable in 12 hours.

(9) replica test

Get 6 parts of same batch samples, press the preparation of need testing solution preparation method, sample introduction 10 μ l record peak area value respectively, calculate content, the results are shown in Table 21.

Table 21 sample replica test result

Result of the test shows: sample RSD is 0.77%, less than 2%, so repeatability is good.

(10) average recovery test

Get the about 1.5g of sample powder of 9 parts of known content, accurately weighed, be prepared into medicinal liquid according to the medicinal liquid preparation method, and be prepared into test sample by the preparation method of above-mentioned need testing solution, add respectively three kinds of various dose (0.058mg/ml of Sennoside A reference substance solution, 12ml, 15ml, 18ml), parallel three parts, sample introduction 10 μ l measure peak area, calculate recovery rate the results are shown in Table 22.

Table 22 average recovery result of the test

Result of the test shows: all between 90～96%, its average recovery rate is that 91.99%, RSD is 1.92% to the average recovery measurement result of 9 experiments, less than 2%, so application of sample reclaims well.

(11) sample size is measured

Accurate reference substance solution and each 20ul of need testing solution of drawing measures according to above-mentioned chromatographic condition respectively, and record spectrum peak area value also calculates content, the results are shown in Table 23.

Table 23 assay result

By the assay result as can be known: the Sennoside A average content of three batch samples is 5.614mg/g, and tentative this product medicinal liquid Sennoside A content is not less than 4.490mg/g.

2 magnolol, honokiol assay

(1) instrument and reagent

Shimadzu LC-20A high performance liquid chromatograph, Sai Duolisi electronic balance (Bp211DAG Germany Sartorius), the multiplex vacuum pump of SHD-III type circulating water type (high and new technology industrial development zone, Baoding sunlight Science ﹠ Teaching Instrument factory), ultrasonic cleaner CX-100(Beijing armarium two factories).

Methanol is chromatographically pure, and water is double distilled water, and all the other reagent are analytical pure.

Magnolol reference substance (lot number: 110729-200309, Nat'l Pharmaceutical ﹠ Biological Products Control Institute), honokiol reference substance (lot number: 0730-200206, Nat'l Pharmaceutical ﹠ Biological Products Control Institute).

(2) chromatographic condition

With reference to " relevant regulations is measured under item of Chinese pharmacopoeia (2010 editions).

Chromatographic column: Comatex-C ₁₈Post (5 μ m, 250mm * 4.6mm)

Mobile phase: methanol-water (78:22)

Flow velocity: 1ml/min;

Detect wavelength: 294nm;

Column temperature: 35 ℃.

(3) preparation of reference substance solution

Get magnolol, the honokiol reference substance is an amount of, and is accurately weighed, add methanol and make respectively the solution that every 1ml contains magnolol 28ug, honokiol 30ug, and get final product.

(4) preparation of need testing solution

The accurate medicinal liquid 0.5ml that draws volatilizes, and residue adds dissolve with methanol and standardize solution is put in the 25ml volumetric flask, and get final product.

(5) preparation of negative sample solution

Get the Cortex Magnoliae Officinalis negative sample, with reference to the preparation of test sample preparation method, and get final product.

Reference substance, reference substance are negative, test sample is negative and the high-efficient liquid phase chromatogram of test sample is seen accompanying drawing 9～Figure 12.

(6) linear relationship is investigated

Accurate honokiol reference substance solution (30.512ug/ml), magnolol reference substance solution (28.320ug/ml) sample introduction 1ul, 2ul, 4ul, 6ul, the 8ul of drawing measures peak area, the results are shown in Table 24,25.

The table 24 honokiol range of linearity is investigated the result

The table 25 magnolol range of linearity is investigated the result

Take honokiol, magnolol reference substance sample size as abscissa, its corresponding peak area value is the vertical coordinate curve plotting respectively, gets the honokiol regression equation and is: Y=3E+06X-1578, R ²=1, show that honokiol has good linear relationship between 0.03051～0.24410ug.The regression equation of magnolol is: Y=6E+06X-9627, R ²=0.9998, show that magnolol has good linear relationship between 0.02832～0.22656ug.

(7) precision test

The above-mentioned reference substance solution 10ul of accurate absorption repeats sample introduction 6 times, and the record peak area the results are shown in Table 26,27.

Table 26 honokiol Precision test result

Table 27 magnolol Precision test result

Result of the test shows: RSD is all less than 2%, so instrument precision is good.

(8) stability test

Get same reference substance and same need testing solution, respectively at 0,2,4,8,12 hour sample introduction 10 μ l, the record peak area value the results are shown in Table 28,29,30.

Table 28 honokiol reference substance stability test result

Table 29 magnolol reference substance stability test result

Table 30 test sample stability test result

Result of the test shows: reference substance and test sample RSD are all less than 2%, so reference substance solution and test sample solution are stable in 12 hours.

(9) replica test

Get 6 parts of same batch samples, press respectively the preparation method preparation of need testing solution, sample introduction 10 μ l, the record peak area calculates content, the results are shown in Table 31.

Table 31 test sample replica test result

Result of the test shows: sample RSD is less than 2%, so repeatability is good.

(10) average recovery test

Get the about 1.5g of sample powder of 9 parts of known content, accurately weighed, be prepared into medicinal liquid, be prepared into test sample by the test sample preparation method, add respectively honokiol reference substance solution (30.512ug/ml), three of magnolol reference substance solution (28.320ug/ml) are not commensurability, parallel three parts, sample introduction 10 μ l, measure peak area value, calculate recovery rate the results are shown in Table 32,33.

Honokiol average recovery result of the test in table 32 test sample

Magnolol average recovery result of the test in table 33 test sample

Result of the test shows: 3 kinds of varying levels of honokiol, and all between 90～96%, its average recovery rate is that 92.99%, RSD is 1.26% to the average recovery measurement result of 9 experiments, it is good to show that application of sample reclaims;

3 kinds of varying levels of magnolol, all between 87～95%, its average recovery rate is that 90.87%, RSD is 1.67% to the average recovery measurement result of 9 experiments, it is good to show that application of sample reclaims.

(11) sample size is measured

Accurate reference substance solution and each 10ul of need testing solution of drawing measures according to above-mentioned chromatographic condition respectively, and record chromatographic peak area value is calculated content, the results are shown in Table 34.

Table 34 assay result

By measurement result as can be known: the honokiol average content of three batch samples is 46.676mg/g, and the magnolol average content is 52.290mg/g, so tentative this product medicinal liquid honokiol content is not less than 37.340mg/g, Determination of Magnolol is not less than 41.830mg/g.

The preparation of embodiment 3 medicine patch of the present invention

1) ratio in 3:1.5:1 takes by weighing respectively Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, be ground into coarse powder, add 75% alcohol dipping 6 hours, carry out the normal pressure percolation, the percolate natural subsidence is got supernatant and is evaporated to without the alcohol flavor in 40 ℃, be mixed with the medicinal liquid of 1g/ml with 25% ethanol, the Natrii Sulfas that adds medicinal liquid weight 10～25% makes its dissolving, adds medicinal liquid weight 2% azone again, namely gets the medicinal liquid that concentration is primary crude drug meter 1g/ml;

2) take by weighing Radix Et Rhizoma Rhei, Natrii Sulfas in 1: 1 ratio, be ground into fine powder, mix homogeneously, sterilization namely gets medicated powder;

3) medicated powder is sealed up for safekeeping in the breathability sticker, liquid medicine filling is prepared into patch.

Wherein, the drug loading of medicinal liquid is 1-3ml, and concentration is counted 1-2g/ml with primary crude drug, and the medicated powder drug loading is 1-3g.

The preparation of embodiment 4 medicine unguentum of the present invention

Get Radix Et Rhizoma Rhei 300g, Cortex Magnoliae Officinalis 150g, Fructus Aurantii Immaturus 100g, Natrii Sulfas 10g, directly beat powder, add matrix composition and become externally used paste.

The preparation of embodiment 5 medicine patch of the present invention

Get Radix Et Rhizoma Rhei 100g, Cortex Magnoliae Officinalis 50g, Fructus Aurantii Immaturus 50g, Natrii Sulfas 5g, be prepared into patch by the method for embodiment 3.

The preparation of embodiment 6 medicine patch of the present invention

Get Radix Et Rhizoma Rhei 500g, Cortex Magnoliae Officinalis 300g, Fructus Aurantii Immaturus 150g, Natrii Sulfas 40g, be prepared into patch by the method for embodiment 3.

Below prove beneficial effect of the present invention by concrete pharmacodynamics test.

Test example 1 medicine of the present invention rushes down lower experiment

1. instrument, reagent and animal

Instrument mouse stomach device, self-control mouse metabolism cage (utility model patent, the patent No.: CN201146735), filter paper, timing watch, electronic balance (German Sartorius company).

Medicine compound diphenoxylate (the accurate word of traditional Chinese medicines: H34021501 Hefei joins pharmaceutical Co. Ltd for a long time), Fructus Cannabis Bolus (Hubei promise win victory pharmaceutical Co. Ltd).

Reagent azone (chemical pure CP), sodium sulfide are all produced by the Long Huagongshijichang of Chengdu section, and normal saline is produced by Kelun Pharm Ind Co., Ltd., Sichuan.

Animal health Kunming mouse (cleaning level), male and female half and half, body weight 18～22g is purchased from Sichuan and reaches rich fruit and test animal center, animal production licence number SCXK (river) 2008-24.

2. test place

Chengdu University of Traditional Chinese Medicine's Chinese medicine processing laboratory (three grades of laboratorys of State Administration of Traditional Chinese Medicine); Herbal pharmacology laboratory animal observation ward of pharmaceutical college of Chengdu University of Traditional Chinese Medicine, credit number: the real moving pipe in river makes No. 15.

3. statistical method

Experimental data is with mean+SD

Expression, the one factor analysis of variance method that the significance test of group difference uses SPSS software to provide is analyzed.

4. the preparation of medicine

Preparation for the reagent thing takes by weighing respectively Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, each 300g of Fructus Aurantii Immaturus, 150g, 100g, is ground into coarse powder, and according to medicinal liquid preparation technology preparation, the medicinal liquid that is formulated as respectively basic, normal, high three kinds of concentration (0.5g/ml, 1g/ml, 2g/ml) is an amount of.Take by weighing Radix Et Rhizoma Rhei, each 50g of Natrii Sulfas, be ground into fine powder, mix homogeneously is made respectively the sticker of basic, normal, high (0.25g, 0.5g, 1g) Three doses.

The preparation compound diphenoxylate of model drug, every hydrochloric diphenoxylate 2.5mg gets that to be mixed with in right amount 0.5mg/ml for subsequent use the time spent ^[19,20]

The preparation Fructus Cannabis Bolus of positive drug is got 30g during use, it is for subsequent use to be mixed with 0.5g/ml.

The preparation Radix Acaciae senegalis 10g of prepared Chinese ink adds water 80ml, boils to solution transparently, gets active carbon 5g, adds mentioned solution and boils 3 times, after the solution cold, adds water and is settled to 100ml, and 4 ℃ of Refrigerator stores shake up before use.

5. process of the test

5.1 grouping

Get 50 of healthy kunming mices, male and female half and half are divided into 5 groups at random by body weight, and namely model group, low dose group, middle dosage group, high dose group and the positive are organized 10 every group.

5.2 depilation

To mice processings of losing hair or feathers, 8% sodium sulfide is applied to mouse web portion before the experiment, behind the 1min, with clean distilled water cleaning skin, normally raises 24h.

5.3 experimental technique

Water 12h is can't help in fasting before the experiment, and each organizes respectively that gavage gives compound diphenoxylate 50mg/kg, behind the 1h, each is organized rapid gavage and gives prepared Chinese ink and each self administration of medication and sub-cage rearing, model group gives blank patch, and basic, normal, high dosage group gives relative medicine, positive group gavage Fructus Cannabis Bolus medicinal liquid 5g/kg ^[21]

5.4 observation index

Begin timing from administration, Continuous Observation 10h records each animal and arranges first melena time and 10h accumulation defecation number.

6. result of the test

The statistics mice is arranged melena time and 10 hours accumulation defecation number first, the results are shown in Table 35.

Table 35 rushes down lower experimental result table

Annotate: compare P＜0.05 ﹡, P＜0.01 ★ with model group

Known by the results of analysis of variance that (1) melena time P is arranged in Normal group and model group contrast first〉0.05,10h accumulation defecation is counted P＜0.01, and namely for arranging first the melena experiment, modeling is unsuccessful, but modeling success for accumulation defecation number.

(2) respectively basic, normal, high dosage group and positive group are compared with model group, known by the results of analysis of variance: concerning arranging first the melena time, basic, normal, high dosage group has significant difference, P＜0.01, and positive group also has significant difference P＜0.01.

For 10h accumulation defecation number, low dose group and model group contrast do not have statistical significance, and middle dosage group has significant difference, P＜0.05, and high dose group and positive group have significant difference P＜0.01.

(3) respectively each administration group and Normal group are compared, known by variance analysis, for arranging first melena, low dose group P=0.437 0.05, there is not statistical significance, middle dosage group, high dose group and positive group have significant difference, P＜0.01.

Test example 2 Pharmaceutical sausage Promoting Experiments of the present invention

1. instrument, reagent and animal

Instrument mouse stomach device, operating scissors, ophthalmic tweezers, ruler, electronic balance (German Sartorius company).

Reagent azone (chemical pure CP), sodium sulfide are produced by the Long Huagongshijichang of Chengdu section, and normal saline is produced by Kelun Pharm Ind Co., Ltd., Sichuan.

Animal health Kunming mouse (cleaning level), male and female half and half, body weight 18～22g is purchased from Sichuan and reaches rich fruit and test animal center.Animal production licence number SCXK (river) 2008-24.

2. test place

3. statistical method

Experimental data is with mean+SD

Expression, the one factor analysis of variance method that the significance test of group difference uses SPSS software to provide.

4. supply the preparation of reagent thing

For the 4th lower operation in the same joint of the preparation of reagent thing.

5. process of the test

5.1 grouping

Get 50 of healthy kunming mices, male and female half and half are divided into 6 groups at random by body weight, and namely model group, low dose group, middle dosage group, high dose group and the positive are organized 10 every group.

5.2 depilation

To the mice processing of losing hair or feathers, 8% sodium sulfide is applied to mouse web portion before the experiment, behind the 1min, cleans skin with distilled water, normally raise 24h.

5.3 experimental technique

Water 12h is can't help in fasting before the experiment, and each is organized gavage and gives compound diphenoxylate 5mg/kg, behind the 30min, each is organized rapid gavage and gives prepared Chinese ink and each self administration of medication, model group gives blank patch, and basic, normal, high dosage group gives corresponding medicine, positive group gavage Fructus Cannabis Bolus medicinal liquid 5g/kg ^[22-24]

5.4 observation index

Put to death immediately behind the administration 25min, cut open the belly to take out and go back to section to whole small intestinals of ileocecus from pylorus, cut off mesentery, small intestinal is stretching gently, be tiled on the blank sheet of paper, measure and record prepared Chinese ink propelling length, calculate the prepared Chinese ink ink propulsive rate.

6. result of the test

Small intestinal total length, the prepared Chinese ink of statistics mice advance length and propelling rate at enteral, the results are shown in Table 36.

Table 36 intestinal advances as a result table

Annotate: with model group contrast, P＜0.05 ﹡, P＜0.01 ★

(1) known by the results of analysis of variance, Normal group and model group contrast, so P=0.033＜0.05 is the modeling success.

(2) respectively basic, normal, high dosage group and positive group are compared with model group, known by the results of analysis of variance: low dose group P=0.096 0.05, there is not statistical significance, middle dosage group P=0.024＜0.05, significant difference is arranged, and high dose group and positive group have significant difference P＜0.01.

(3) respectively each administration group and Normal group are compared, known by variance analysis, each administration group does not all have statistical significance.

Medicine patch of the present invention can shorten the first defecation time of mice with constipation really, and effect is better than the Fructus Cannabis Bolus group, and middle and high dosage group also can shorten first defecation time of normal mouse; Medicine patch of the present invention also can increase accumulation defecation number and the small intestinal recommendation rate of mice with constipation, proves that through the test of pesticide effectiveness this patch is definite to the mice with constipation discharge function.

Below prove safety of the present invention by skin irritation and sensitivity test.

Test example 3 medicine skin irritation tests of the present invention

1. instrument, reagent and animal

The instrument operating scissors, electronic balance (German Sartorius company).

Reagent sodium sulfide (Long Huagongshijichang of Chengdu section production).

The animal rabbit, male and female half and half, female without pregnant, body weight 1800 ± 100g, animal production licence number SCXK (river) 2008-24 is purchased from Sichuan and reaches rich fruit and test animal center.

2. test place

3. statistical method

Relatively adopt the χ of SPSS software between group ²Check.

4. for the reagent thing

Test with pilot product.

5. single-dose skin irritation test method

Get 8 rabbit, male and female half and half are divided into the intact skin group, and the damaged skin group, adopts to contrast about consubstantiality self and tests by 4 every group.Give family's rabbit back spinal column both sides unhairing, the depilation area is about 7cm * 7cm.24h checks whether skin of unhairing is injured, and injured rabbit is unfit to do the toxicity test of intact skin after the unhairing.

The making of damaged skin: adopt the disposable syringe syringe needle to mark " # " shape otch at depilation district sterilization skin, each about 5cm of length and width, the degree of depth is 2～3mm, not damage subcutaneous tissue and oozing of blood to be arranged as degree ^[25]

Unhairing district, the animal left side invention medicine patch that loses money in a business, blank patch is pasted in unhairing district in right side.Behind the administration 8h, remove residual tested material with warm water, remove after the tested material 1h, 24h, 48h, 72h perusal and record dispenser place and have or not the situations such as erythema, edema, by " skin irritation reaction standards of grading " ^{[26,27 ］}Mark, calculate each treated animal reaction mean scores.According to " skin irritation intensity evaluation standard " ^{[26,27 ］}Skin irritation intensity is estimated." skin irritation reaction standards of grading " see Table 37, and " skin irritation intensity evaluation standard " sees Table 38, and experimental result sees Table 39.

Reaction mean scores=(erythema forms total points+edema and forms total points)/number of animals

Table 37 skin irritation reaction standards of grading

Table 38 skin irritation intensity evaluation standard

6. multiple dosing skin irritation test method

Method is with a medicine-feeding test, but administration every day 1 time, 1 week of successive administration.Remove after the tested

material

1,24,48, the 72h perusal, the average response score value of tested material and substrate respectively organized in record, marks by " skin irritation reaction standards of grading ", calculates each treated animal reaction mean scores.According to " skin irritation intensity evaluation standard " skin irritation intensity is estimated ^[28], experimental result sees Table 39.

7. result of the test

(1) single-dose irritation test result

The medication of table 39 medicine patch single of the present invention is to skin irritant average response score value

1. the average response score value difference not statistically significant of medicine patch of the present invention and blank patch single administration (P〉0.05).

2. to the intact skin treated animal, the average response score value of medicine patch of the present invention and blank patch all＜0.5 minute, show that both are to the normal skin nonirritant;

3. to the damaged skin treated animal, the average response score value of medicine patch of the present invention after medication 1h and 24h all 〉=0.5 minute, but＜2.99 minutes, show that medicine patch of the present invention has slight zest to damaged skin, behind the administration 72h, zest disappears, the average response score value of blank patch 1h time 〉=0.5 minute after medication, but＜2.99 minutes, show that blank patch has slight zest to damaged skin, behind the administration 48h, zest disappears.

(2) multiple dosing irritation test result

Table 40 medicine patch multiple dosing of the present invention is to the average response score value of skin irritation

1. the average response score value difference not statistically significant of medicine patch of the present invention and blank patch multiple dosing (P〉0.05).

2. to the intact skin treated animal, the average response score value behind medicine patch of the present invention and the blank patch medication 1h all 〉=0.5 minute, show that both have slight zest to normal skin, zest disappears behind the medication 24h;

3. to the damaged skin treated animal, the average response score value of medicine patch of the present invention after medication 1h and 24h all 〉=0.5 minute, but＜2.99 minutes, show that medicine patch of the present invention has slight zest to damaged skin, behind the administration 72h, this zest disappears; The average response score value of blank patch is after medication during 1h〉0.5 minute, but＜2.99 minutes, showing that blank patch has slight zest to damaged skin, behind the administration 48h, this zest disappears.

Test example 4 medicine skin hypersensitivity experiments of the present invention

1. instrument, reagent and animal

The medicine DNFB

Reagent sodium sulfide is produced by the Long Huagongshijichang of Chengdu section.

The animal rabbit, male and female half and half, female without pregnant, body weight 1800 ± 100g is purchased from Sichuan and reaches rich fruit and test animal center.Animal production licence number SCXK (river) 2008-24.

2. test place

3. statistical method

Relatively adopt SPSS software between group, adopt χ ²Check.

4. for the reagent thing

Test with pilot product.

5. test method

(1) 18 of rabbit are got in the preparation of grouping and skin, are divided at random 3 groups (blank group, medicine patch group of the present invention, positive groups), and 6 every group, male and female half and half.24h is with family's rabbit back spinal column both sides unhairing before administration, and the depilation area is 7cm * 7cm.

(2) sensitization contact is got tested material and is given depilation district, animal left side, and medicine patch group of the present invention gives medicine patch of the present invention, and blank group gives blank patch, positive controls gives 1%2,4-dinitrochlorobenzene 0.1m, continues 8h, the 7th day and the 14th day in kind repeats 1 time, and administration is 3 times altogether ^[29,30]

(3) excite be contacted with last to tested material sensitization after 14d, tested material is imposed on respectively the depilation district, right side of corresponding animal, remove tested material behind the 8h, at once observe, and again observe the skin allergy situation in 24h, 48h, 72h, by " standards of grading of skin allergy degree " ^{[26,27 ］}Mark, calculate each treated animal reaction mean scores (seeing Table 41).Note observing animal whether the serious whole body allergy such as asthma, shock or astasia are arranged.According to " skin hypersensitivity evaluation criterion " ^{[26,27 ］}Infer sensitization (seeing Table 42) by the sensitization incidence rate

Sensitivity response incidence rate=have anaphylaxis number of animals/animal number * 100%

The standards of grading of table 41 skin allergy degree

Table 42 hypersensitive evaluation criterion

6. result of the test

Medicine patch of the present invention to the rabbit skin allergy test the results are shown in Table 43, table 44.

Table 43 medicine patch of the present invention is to rabbit sensitization of skin reaction mean scores

Table 44 medicine patch of the present invention is on the impact (%) of rabbit skin sensitization rate

1. blank group has no the skin allergic reaction situations such as erythema, edema;

2. medicine patch group of the present invention only has individual animal namely to be carved with the moderate erythema reaction, and the sensitization rate is that 30%, 48h afterreaction disappears;

3. the skin allergic reaction situations such as obvious erythema, edema appear in positive controls, and the sensitization rate reaches 100%.

The serious whole body allergy such as asthma, astasia or shock all do not appear in 3 groups of rabbit.

From skin irritation and anaphylaxis experimental result as can be known: this medicine has slight zest to the intact skin nonirritant to damaged skin, but can die away; Skin there is slight sensitization, but can dies away, proved that the safety of patch of the present invention is higher.

List of references:

[1] Huang Taikang, the chief editor. conventional Chinese medicine composition and pharmacology handbook [M]. Beijing: Chinese Medicine science and technology publishing house, 1994:227

[2] Yang Shiyou, Huang Shifu, Tian Jun etc. Different Extraction Method is rushed down the impact [J] of lower composition on Radix Et Rhizoma Rhei. Anhui medicine, 2004,9(2): 9092.

[3] Li Min, Liu Yu, Li Lixia. rhubarb formula granules Assay of Total Anthraquinone methodological study [J]. Chengdu University of Traditional Chinese Medicine's newspaper, 2006,9 (1): 4750.

[4] Ma Rong, Zhang Xueju. liquid chromatography contains amount comparing analysis [J] to Sennoside A in the Radix Et Rhizoma Rhei of the different places of production. Chinese patent medicine, 2008,30 (10): 14891490.

[5] Zheng Zhihua, Zhu Chen dip in the content [J] that .HPLC measures Extraction process of Rhubarb product Sennoside A. Chinese crude drug, 2004,27 (12): 950951.

[6] grandson wears, Li Min, the content [J] of Sennoside A and sennoside B in the little mensuration rhubarb medicinal materials such as methods such as the .HPLC of grade of poplar and the decoction pieces more. and Chengdu University of Traditional Chinese Medicine reports, and 2008,31 (3): 5153.

[7] Chinese Pharmacopoeia Commission. the Pharmacopoeia of the People's Republic of China 2010 editions an one [M]. Beijing: the .2010. of Chemical Industry Press

[8] Yang Shiyou, Huang Shifu, Tian Jun etc. Different Extraction Method is rushed down the impact [J] of lower composition on Radix Et Rhizoma Rhei. Anhui medicine, 2004,9 (2): 9092.

[9] Liu Siyu, Liu Xiaolong, Zhang Quanyu etc. the basic procedure of detection of drugs percutaneous permeation [J]. northwest pharmaceutical journal, 2009,24 (6): 511513.

[10] Su Xiaomei, Wang Fang, Wang Guohua etc. the preparation of Rhizoma Zingiberis Recens gel adhesive and the research of Transdermal Absorption [J] thereof. Jiangxi Chinese medicine, 2009,40 (1): 6162.

[11] Wang Wenyue, Yun Liuhong, Wang Rui etc. the prescription of felodipine one metoprolol compound transdermal patch and technical study [J]. China Dispensary, 2008,19 (16): 12361239.

[12] Liu Chunhai, Yang Yonghua, Zhang Shuihan. the Shangtong Tincture concentration of alcohol is on the impact [J] of paeonol Transdermal absorption. Chinese patent medicine, 2001,23 (10): 710711.

[13] Liu Yi, Zhou Guanhuai, Chen Hongzhuan. ethanol is on the impact [J] of tetracaine gel transdermal and local anesthetic action. Chinese Clinical pharmacology and therapeutics, 2003,8 (5): 523525.

[14] Wu Qiangdong, woods are luxuriant, a flower bud. the composition pharmacology of Natrii Sulfas and new [J] .Strait Pharmaceutical Journa, 2009,21 (1): 135. of using

[15] woods Jianyang, Li Hua, Li Jiayu. the impact [J] that azone absorbs Vinpocetine transdermal. mathematical medicine magazine 2002,15 (4): 354355.

[16] in great waves, Tang Lei, Kang Li. the azone consumption is on the impact [J] of the stripped Transdermal absorption of monobenzone emulsifiable paste. West China pharmaceutical journal, 2010,25 (2): 151153.

[17] Yu Linbo, Xie Xingqin. menthol, azone, propylene glycol use in conjunction are on the impact [J] of Sumei fat-reducing plaster percutaneous rate. Heilungkiang medicine 2008,21 (4): 8889.

[18] Wei Li, Cai Zhenzhen, Xu Lianying. the experimental study of Percutaneous Absorption of Ferulic Acic [J]. Chinese patent medicine, 1998,20, (6): 41～42.

[19] Yu Youming, Liang Xueyan, Wang Rujun. the experimentation [J] of QI invigorating relaxing bowel defecating feces excretion. the new traditional Chinese medical science, 2009,41 (12): 100101.

[20] be coated with jasmine, high ferro is auspicious, Huang Li. the experimentation [J] that the happy year square tube of crane just acts on. and Chinese Chinese medicine science and technology, 2009,16 (1): 1617.

[21] Feng Min. American lotus holds function of relaxing bowel research [J]. Industrial and Commercial University Of Chongqing's journal (natural science edition), 2008,25 (4): 416418.

[22] woods is strong, Zhang Rongbiao, Liu Shaojuan. the research of Effect of Catharsis of Qingrun Tea in Mice [J]. and preventive medicine forum, 2007,13 (3): 207208.

[23] Wang Jinping, Ceng Ming, Liu Yuping. Shu Tongfang is to the functions of loosening bowel relieving constipation [J] of mice. Tropical China medical science, 2010,10 (12): 15081509.

[24] Wang Yingjun, Sun Yinglian, Tang Wei. the experimentation of Ease-constipation Action of Capsulae Aloes [J]. special product research, 2007,36 (2): 3638

[25] Yu Baohua, Sheng Guorong. compound lidocaine emulsifiable paste is observed [J] to the guinea pig skin irritant experiment. clinical rational drug use, 2010,3 (5): 4849.

[26] Xie Xiuqiong. new product of Chinese medicine exploitation and application ［ M ］ .2 version. Beijing: People's Health Publisher, 2002:565571.

[27] LiangBing Wen. Chinese medicine percutaneous drug-delivery preparation technology ［ M ］. Beijing: Chemical Industry Press, 2006:171244.

[28] Yu Baohua. dexamethasone urea frost percutaneous drug delivery safety research [J]. clinical rational drug use, 2010,3 (19): 1011.

[29] Wang Qingling, Zhu Mingquan, Yan Zhouping. preparation and the dermatoxicology experimental study [J] thereof of the yellow coagel preparation of compound recipe. the bright traditional Chinese medical science, 2010,25 (10): 17981800.

[30] Zhou Fan, Feng Shibin, Meng Qingjuan. the dermatologic safety research [J] of Chinese medicine compound transdermal patch. herding and forage science, 2010,31 (3): 5254.

Claims

1. pharmaceutical composition that is used for the relieving constipation fat-reducing is characterized in that: it is to be the topical agent that crude drug is prepared from by Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas.

2. pharmaceutical composition according to claim 1, it is characterized in that: the weight proportion of described crude drug is:

3. pharmaceutical composition according to claim 2, it is characterized in that: the weight proportion of described crude drug is:

4. the described pharmaceutical composition of any one according to claim 1-3, it is characterized in that: it is to be active component by the protogenic medicinal powder of Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus, Natrii Sulfas or extractive with organic solvent, adds the medicament that pharmacy external preparation adjuvant commonly used or complementary composition are prepared from.

5. pharmaceutical composition according to claim 4, it is characterized in that: described topical agent is tincture, unguentum, lotion, liniment, cataplasma, patch.

6. pharmaceutical composition according to claim 5 is characterized in that: comprise in the described patch and sealing up for safekeeping in the medicated powder of breathability sticker and the medicinal liquid of fill; Wherein, the drug loading of medicinal liquid is 1-3ml, and concentration is counted 1-2g/ml with primary crude drug, and the medicated powder drug loading is 1-3g.

7. pharmaceutical composition according to claim 6, it is characterized in that: Sennoside A content is not less than 4.490mg/g in the described medicinal liquid; Be not less than 37.340mg/g with Determination of Magnolol in the medicinal liquid, Determination of Magnolol is not less than 41.830mg/g.

8. method for preparing pharmaceutical composition claimed in claim 1, it comprises the steps:

A, weighting raw materials;

C, get Radix Et Rhizoma Rhei, Natrii Sulfas, be ground into fine powder, fine powder, middle powder, or coarse powder, mix homogeneously, sterilization namely gets medicated powder;

9. preparation method according to claim 8, it is characterized in that: it comprises the steps:

A, weighting raw materials;

B, take by weighing respectively Radix Et Rhizoma Rhei, Cortex Magnoliae Officinalis, Fructus Aurantii Immaturus in the ratio of 3:1.5:1, be ground into coarse powder, add 75% alcohol dipping 6 hours, carry out the normal pressure percolation, the percolate natural subsidence is got supernatant and is evaporated to without the alcohol flavor in 40 ℃, be mixed with the medicinal liquid of 1g/ml with 25% ethanol, the Natrii Sulfas that adds medicinal liquid weight 10～25% makes its dissolving, adds medicinal liquid weight 2% azone again, namely gets the medicinal liquid that concentration is primary crude drug meter 1g/ml;

C, take by weighing Radix Et Rhizoma Rhei, Natrii Sulfas in 1: 1 ratio, be ground into fine powder, mix homogeneously, sterilization namely gets medicated powder;

10. the purposes of the described pharmaceutical composition of claim 1-7 any one in the external used medicine of preparation relieving constipation fat-reducing.