CN103919967A

CN103919967A - Dracaena cochinchinensis resin compound and application thereof in preparation of medicines for treating trauma

Info

Publication number: CN103919967A
Application number: CN201410133145.9A
Authority: CN
Inventors: 张荣平; 余晓玲; 赵荣华; 赵昱; 巫秀美; 于浩飞; 石聪; 李雷; 袁小淋; 郝艳兵; 张琦; 郑园园; 张兰春
Original assignee: Individual
Current assignee: Individual
Priority date: 2014-04-03
Filing date: 2014-04-03
Publication date: 2014-07-16
Anticipated expiration: 2034-04-03
Also published as: CN103919967B

Abstract

The invention provides a compound preparation which comprises dracaena cochinchinensis resin or daemonorops draco resin and other components and has the effects of trauma treatment, tissue repair, angiitis treatment and the like. The compound preparation is characterized in that the ratio of the dracaena cochinchinensis resin or the daemonorops draco resin to borneol is 9:1, and the ratio of the dracaena cochinchinensis resin or the daemonorops draco resin to berberine to borneol is 9:4:1, or the ratio of the dracaena cochinchinensis resin or the daemonorops draco resin to the berberine to the borneol is 1:1:0.15. The invention also provides a preparation method of the compound preparation and application of the dracaena cochinchinensis resin or daemonorops draco resin compound in the preparation of compound gels, liniment, cataplasm, adhesive bandages (such as patches) and other preparations. Compared with two positive medicines (Madecassol, jingwanhong soft plaster), the dracaena cochinchinensis resin or daemonorops draco resin compound preparation has the obvious effects of promoting wound healing and tissue repair. The acute toxicity test and the long-term toxicity test show that the preparation is safe to use.

Description

Sanguis Draxonis compound recipe and more create the application in medicine in preparation treatment

Technical field:

The invention belongs to technical field of pharmaceuticals, particularly, relate to the drug compound preparation that Sanguis Draxonis or Sanguis Draxonis are main component, prepare the method for this compound preparation and the application in the medicine of preparation treatment more wound or tissue repair thereof.

Background technology:

Sanguis Draxonis is warm in nature, flat, sweet in the mouth, salty, nontoxic, enter blood system, return lung, spleen, kidney three warps, there is the remarkable efficacy such as blood circulation promoting and blood stasis dispelling, astringing to arrest bleeding, hard masses softening and resolving, promoting tissue regeneration and ulcer healing, be usually used in inside and outside all sections, as diseases such as: traumatic injury, blood stasis, hemorrhage, carbuncle ulcer, the disunion of a specified duration of skin ulcer face, be many ancient prescriptions and the important composition medical material through proved recipe, be called " panacea of invigorating blood circulation " prompting Sanguis Draxonis and there is repair in trauma effect.The medicinal Sanguis Draxonis that China is used the earliest " comes from the Western Regions ", and its Original plant is the Agavaceae plant Folium Dracaenae cambodianae (Dracaena draco) that originates in West Asia and north African.Bright clear since, Indonesia and Malaysian Palmae Daemonorops plant Sanguis Draxonis (Daemonorops dracoBL.) red resin that produces become the main source of the medicinal Sanguis Draxonis of China, be " Sanguis Draxonis " (the resin from Daemonorops draco that the Pharmacopoeia of the People's Republic of China 2010 editions records, RDD, claims Sanguis Draxonis herein).Last century the seventies, famous botanist Cai Xitao professor finds to secrete the Agavaceae dracaena plant Dracaena cochinchinensis (Dracaena cochinchinensis Lour.S.C.Chen) of red resin in Yunnan Province, successfully develop domestic Dragon Blood, listed " national drug standards " promulgated by the ministries or commissions of the Central Government (WS3-082 (Z-016)-99 (Z)) in 1999, called after Sanguis Draxonis (the resin from Dracaena cochinchinensis, RDC), thus opened up approach for the production domesticization of famous and precious Chinese medicine Resina draconis.But it is different that domestic Sanguis Draxonis (RDC) and import Sanguis Draxonis (RDD, i.e. Sanguis Draxonis) plant section belongs to, there were significant differences for chemical composition.At present Sanguis Draxonis and Sanguis Draxonis (that is: Sanguis Draxonis) be all clinically for blood circulation promoting and blood stasis dispelling, promoting tissue regeneration and ulcer healing, but two kinds of Sanguis Draxonis lack basic research more creating aspect the medicinal effects of field, and whether Sanguis Draxonis can substitute Sanguis Draxonis and also have no report for wound aspect more.For the chemical constitution study of Sanguis Draxonis, the result obtaining from different Original plants is not quite similar.The complexity of Sanguis Draxonis chemical composition makes the particularly difficulty of determining of its active component.At present with monomeric form be confirmed to be more create active component only from Euphorbiaceae Fructus Crotonis platymiscium Croton salutaris, C.lechleri produces the alkaloid taspine being separated in Sanguis Draxonis.Domestic scholars thinks that to the chemical research of Sanguis Draxonis its main component is flavonoid and phenols, and import Sanguis Draxonis mainly contains polyphenolics and ter penoids, but both are neither containing taspine.The modern pharmacology research of Sanguis Draxonis concentrates on its using blood system always.Zoopery shows, Sanguis Draxonis can reduce packed cell volume, whole blood viscosity and plasma viscosity, can anticoagulant, and blood vessel dilating, improves microcirculatory blood flow, embodies the effect of invigorating blood circulation; Can shorten clotting time, blood plasma recalcification time and euglobulin lysis time, and there is heparin resistance, can be used for hemostasis simultaneously.The two-ways regulation mechanism of invigorating blood circulation-stop blooding of Sanguis Draxonis has been explained its therapeutical effect to coronary heart disease, cerebral ischemia, metrorrhagia etc. and blood coagulation-haemolysis balance destruction diseases related.Tao Yi etc. has compared in the body of Sanguis Draxonis and Sanguis Draxonis, In Vitro Anti platelet aggregation, and result shows both remarkable anticoagulations, and Sanguis Draxonis effect is slightly excellent.Another remarkable drug effect---expelling pus and promoting granulation---of Sanguis Draxonis is also confirmed in a large amount of clinical practices.For a long time, Sanguis Draxonis is widely used in soft tissue contusion, decubital ulcer, pressure ulcer, diabetic foot ulcer, varicosis ulcer of the lower limb and oral ulcer treatment at surgery and department of dermatologry, has obtained good effect.This confirms that Sanguis Draxonis, Sanguis Draxonis stop blooding-invigorate blood circulation dual regulation except having clinically, also has the effect that promotes wound healing.

Both at home and abroad the basic research of wound healing is shown at present, wound healing is divided into three phases: inflammation, tissue form and tissue remodeling.Three phases crossover carries out, and each stage all has specific cytokine and chemokines to participate in, and by the regulating and controlling effect to each stage factor, medicine can accelerated wound healing process and reduced scar tissue and generate.

Sanguis Draxonis is comparatively general in clinical practice, common taking peroral dosage form as main on market.Its main dosage form is capsule and dispersible tablet etc., and main pharmacological is blood circulation promoting and blood stasis dispelling.But oral formulations has drug effect, the performance time more slowly has zest to gastrointestinal tract, liver first-pass effects etc., treat the normal method that uses Sanguis Draxonis dry powder and other drug to match or use the ointment of Sanguis Draxonis such as wound or decubital ulcer now clinically.So far, in prior art there are no taking Sanguis Draxonis, Sanguis Draxonis as main formula composition Chinese medicine compound for the preparation of the report of the application in the medicine for the treatment of more wound or tissue repair.

Summary of the invention

In order to overcome prior art above shortcomings part, it is poor to drug compliance to the object of the invention is to for patient, make medicine can bring into play soon drug effect for trauma patient, Sanguis Draxonis, Sanguis Draxonis are made to the external preparation such as gel, liniment, cataplasma, wound patch, preparation capable of permeating skin (as patch), not only reduced the toxic and side effects of medicine but also can bring into play general action by part.

In order to realize above-mentioned purpose of the present invention, the invention provides following technical scheme:

A kind of Sanguis Draxonis/Sanguis Draxonis compound recipe, it consists of the Sanguis Draxonis of weight portion: Borneolum Syntheticum 9:1 or Sanguis Draxonis: Borneolum Syntheticum 9:1.

A kind of Sanguis Draxonis compound recipe, it consists of the Sanguis Draxonis of weight portion: berberine: Borneolum Syntheticum 9:4:1 or Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.

With above-mentioned compound recipe as gel, liniment, cataplasma, bandage, preparation capable of permeating skin, patch.

The application that the present invention also provides above-mentioned compound recipe to heal in wound, tissue repair medicine in preparation treatment.

In described application, wherein said medicine is treatment wound, blood stasis wound, suppurative wound, dermatitis, comedo, vasculitis, cirsoid medicine.

The present invention provides a kind of gel simultaneously; get above-mentioned compound recipe of the present invention; its compound medicine weight is 0.15g; with mixing of 2.5mL propylene glycol; add carbomer 9.85g; with triethanolamine adjusting, pH value is 7.0, and gained gel has glossiness, stretchability, uniformity and the centrifugal property of preparation requirement.

With, a kind of liniment, gets above-mentioned compound recipe of the present invention, takes this compound medicine 0.2g, is repeatedly dissolved in propylene glycol 1.0g on a small quantity, makes it fully dissolve the phase as A; Get 4% polyvinyl alcohol 8.0g, add sodium carboxymethyl cellulose 0.2g and polyvinylpyrrolidone 0.5g, 70 DEG C-80 DEG C of water-baths, make the abundant swelling dissolving of adjuvant as B phase; B is added to the A mixing that stirs in mutually, finally add surfactant tween 80 to keep the stability of liniment, liniment is evenly spread upon on the glass plate that scribbles liquid paraffin, 37 DEG C of oven dry in baking oven, the outward appearance of gained liniment has uniformity, stretchability, mobility and film pliability.

And a kind of cataplasma, gets propylene glycol-water mixed solvent of appropriate carbomer 44:56, obtain 20g3% carbomer after swelling, add therein 50g2% aqueous tartaric acid solution, fully mix, for subsequent use; Get above-mentioned compound medicine 6g of the present invention, use 15g propylene glycol: the mixed solvent of 95% ethanol 3:12,50 DEG C of water-baths are dissolved, for subsequent use; Get sodium polyacrylate 4g, CMC-Na2g, aluminium hydroxide 0.3g, EDTA0.5g add in beaker, add 22g glycerol to mix well, for subsequent use; Medicine after dissolving is added to the mixed phase such as sodium polyacrylate-glycerol, after fully mixing, carbomer-tartaric acid is added rapidly wherein, stir rapidly, the local caking of control, then continue to stir 60 minutes, treat mastic deliquescing, granular sensation is coated with while disappearance, and room temperature is placed after 3 days and be get final product.

The present invention relates to Sanguis Draxonis (or Sanguis Draxonis) compound preparation, it consists of Sanguis Draxonis (or Sanguis Draxonis): Borneolum Syntheticum 9:1, Sanguis Draxonis (or Sanguis Draxonis): berberine: Borneolum Syntheticum 9:4:1, Sanguis Draxonis (or Sanguis Draxonis): berberine: Borneolum Syntheticum 1:1:0.15.The application of this compound recipe in preparation more wound or tissue repair medicine, the application in treatment wound, blood stasis wound, suppurative wound, dermatitis, comedo, vasculitis etc. more wound and tissue repair medicine.Described compound recipe and pharmaceutical excipient or adjuvant composition medicine, its dosage form is: the external preparation such as gel, liniment, cataplasma, bandage, preparation capable of permeating skin (as patch).The present invention's research shows: this Sanguis Draxonis compound preparation can promote the healing of mice, rat, rabbit back skin injury wound.Decrustation time, healing time obviously shorten respectively, show that Sanguis Draxonis has the wound healing of promotion, accelerates wound incrustation, decrustation effect; In experiment, also observe Sanguis Draxonis group scar tissue less, it is good that site of injury hair growth recovers.

Brief description of the drawings:

The negative reference substance chromatogram of Fig. 1;

Fig. 2 is compound dragon's blood (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1) gel HPLC chromatogram;

Fig. 3 is 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, lourerin B HPLC standard substance chromatogram;

Fig. 4 is 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one standard curve;

Fig. 5 is lourerin B standard curve;

Fig. 6 is 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one release curve chart;

Fig. 7 is lourerin B release curve chart;

Fig. 8 is that situation is oozed out and wound surface redness, infection, the newborn epidermal hyperplasia situation of edge of wound after administration in rat wound surface surface.8A is Sanguis Draxonis low dose group, and 8B is Sanguis Draxonis high dose group, and 8C is Sanguis Draxonis low dose group, and 8D is Sanguis Draxonis high dose group, and 8E is blank matrix group, and 8F is Mei Jiasi group, and 8G is JINGWANHONG group, and 8H is pure blank group.

Detailed description of the invention:

Further illustrate essentiality content of the present invention with embodiments of the invention below, but do not limit the present invention with this.

Embodiment 1

One, determining of compound dragon's blood/Sanguis Draxonis prescription:

Compound dragon's blood shows the therapeutical effect of certain promotion incrustation, decrustation healing in mice wound test model, and the pharmacological action with Sanguis Draxonis itself with putrefaction-removing granulation-promoting wound healing matches.Rhizoma Coptidis has effect of eliminating fire and detoxication, clearing heat and moistening dryness in this prescription, can make traumatic infection rate reduce, and it is one of compound dragon's blood prescribed treatment effect that minimizing wound initial stage part is oozed out.Because the protective layer due to skin surface in open wound is destroyed, antibacterial is easily invaded and makes wound easily cause infection, reduces wound area so keep wound being had to astriction in not infected situation at wound surface simultaneously.Refrigerant, minimizing pain effect that Borneolum Syntheticum has.Select 1 group of three kinds of ingredients compound dragon's blood medicine (Sanguis Draxonis or Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15), 2 groups of compound dragon's blood medicines (Sanguis Draxonis or Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1), determine the two effect to tissue repair.

Select 40 Kunming mouses, provided by the laboratory animal department of the Chinese Academy of Sciences of Kunming medical university, body weight is at 22～25g, freely drink water and ingest, be divided at random 4 groups, every group of 10 white mice, lumbar injection pentobarbital sodium 40mg/Kg anesthesia, back unhairing, 70% ethanol routine disinfection, paving aseptic towel, cuts the skin that diameter is 1.2cm size at back, dark and fascia muscle, 4 groups give respectively different disposal: 2 groups of positive controls, blank group, 1 group of compound dragon's blood and compound dragon's bloods.Positive controls is coated with the agent of clindamycin lidocaine gel, outer gel substrate, 1 group of the other externally coated compound dragon's blood medicine of drug component, 2 groups of the compound dragon's blood medicines of being coated with of blank group.The equal direct drug injection cotton of all wound surface is smeared, and thickness is about 1mm, at every turn all should be by complete medicine flap coverage, and every group is 1 cage.Wound surface every day, coating 2 times, adopt after the method recording medicine of digital photographing the wound area of 3,6,9,12 days respectively at the number that the white mice of every group was carried out to wound area in postoperative 1 day, 3 days, 6 days, 9 days, 12 days, use image software to analyze wound surface area.Wound area before medication is deducted to wound area after the medication wound area before than medication, and the ratio obtaining is wound surface skin healing percentage rate, and calculates and start decrustation natural law and healing days, as the effect of a tested medicine of evaluation of result.After last medication, monitor rat serum picture next day, then de-neck is put to death, clip wound site skin, be fixed in 10% formalin, specimens paraffin embedding slices, HE dyeing, under microscope, observation and comparison is respectively organized the healing of epidermis reparation, the reparation of dermal collagen fibre bundle, dermic hair follice hypertrophy, corium and the subcutaneous tissue inflammatory cell infiltration of burned rats skin.

The postoperative systemic conditions of all animals is good, without untoward reaction such as diarrhoea, refusing to eat, second day after operation anesthetics remove after spirit normally, diet is normal, after modeling, all wound surface all have hemorrhage, tissue fluid to have to ooze out and surrounding tissue edema.Postoperative coating immediately, within the 2nd day, wound place no blood flows out, the dry hardening of skin trauma mouth, spirit and diet are normal.Start to have thicker crust to cover wound at the 5th day left and right wound mouth, obviously alleviate without wound surface surrounding tissue edema, secretions is less.After the 12nd day, wound surface starts decrustation, and wound is newborn skin, and hair increases thereupon, but still has not capped skin.

Wherein 2 groups of positive control drugs, blank group, 1 group of medicine, medicine between respectively incrustation time, decrustation time and healing rate healing time being organized relatively, the results are shown in Table 1 and table 2.

Table 1 is tested wound area slip

Incrustation time and decrustation time, wound healing time is as shown in table 2.

Table 2 incrustation, decrustation and healing time

Group	The incrustation time (d)	The decrustation time (d)	Healing time (d)
				1	2.75±0.75	9.0±1.0	16.7±3.5

2	5.00±1.00	10.3±1.5	18.5±4.0
				3	2.65±0.65	9.0±3.0	17.2±2.5
4	3.50±0.50	8.5±1.0	15.8±2.0

Histopathology appraisal result

The foundation of wounds in mice pathological section histopathology standards of grading and different pharmaceutical group are to the scoring of wounds in mice pathological section, respectively as following table 3 and table 4.

Table 3 histopathology standards of grading

The impact of table 4 different pharmaceutical group on wounds in mice pathological section histopathology scoring (X ± s ' point) [51]

Group	Epidermal structure	Collagenous fiber bundle	Granulocyte infiltrates
				1	0.5±0.63	0.6±0.8	1.3±0.78
2	1.3±0.65	1.3±0.46	1.9±0.30
				3	0.6±0.45	1.0±0.80	1.3±0.90
4	0.6±0.75	1.6±0.56	1.3±0.78

More than experiment is reached a conclusion, and with blank comparison, compound dragon's blood gel of the present invention, positive control drug have good pharmacological effect aspect wound healing rate.Wherein 2 groups of (Sanguis Draxonis or Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1) effects of compound dragon's blood medicine are more obvious.After finding in test that trauma model is built up, a small amount of tissue of mouse skin and Musclar layer is adhered.The skin hardening of wound site after 2 days, no blood flows out, and diet and mental status are normal.After 5 days, wound site starts to have reddish black blood crusts to cover, and after decrustation, wound area obviously diminishes, and wound site is cerise, around a small amount of hair growth of healing skin.After wound heals completely hair also along with wound area reduce increase.After wound heals completely, hair continued growth continues wound coverage position in wound.

Two, Sanguis Draxonis: Borneolum Syntheticum 9:1 compound gel pharmacodynamic evaluation

Experimental agents Sanguis Draxonis: Borneolum Syntheticum 9:1 compound gel, Sanguis Draxonis: Borneolum Syntheticum 9:1 compound gel and gel-type vehicle, by Kunming, institute of traditional Chinese medicine provides, lot number 20130417;

The anti-scar ointment of positive control drug Mei Jiasi (Madecassol): by Bayer Consumer Care AG, Switzer I produces, lot number TRTO281; JINGWANHONG RUANGAO: by Tianjin Darentang Wanhong Pharmaceutical Co., Ltd, lot number Z12020440.

SD rat, in half and half, 6～8 week age of male and female, body weight 180～220g, is purchased from the laboratory animal department of the Chinese Academy of Sciences of Kunming medical university, the quality certification number: SCXK (Yunnan) 2011-0004.Raising condition: 25～27 DEG C of temperature, 70 ± 10 DEG C of humidity, the animal drinking-water of freely ingesting.

After laboratory animal is weighed, anaesthetize according to body weight (30mg/Kg), after anaesthetizing, be fixed in plank, shave a mao 3cm × 3cm with shaver, carry out wound preparation.

Modeling dips povidone iodine with the test tube that diameter is 2cm, is buckled in rat back (left and right sides outside the about 2cm of spinal column), mentions top layer skin with toothed forceps, and 45 ° and contact skin, shear, dark and subcutaneous.Successful modeling rat is divided into 6 groups at random, 7 every group, raises separately.

First group of administration is Sanguis Draxonis low dose group, and second group is Sanguis Draxonis high dose group, and the 3rd group is Sanguis Draxonis low dose group, the 4th group is Sanguis Draxonis high dose group, and the 5th group is blank matrix group, and the 6th group is Mei Jiasi group, the 7th group is JINGWANHONG group, and the 8th group is pure blank group.Respectively organize administration every day by said medicine, gauze covers, and medical adhesive tape is wrapped up fixing.Every day wound is retouched to meter simultaneously, take pictures.

Have drawn from and within the 1st, 3,7,14 days, carry out animal after modeling and draw materials, clip edge of wound tissue, if wound subsidiary crust outward is taken off crust simultaneously.

The preparation tissue of tissue slice is fixed through 4% paraformaldehyde, after the PBS eluting of 0.1M three times, carries out successively gradient alcohol dehydration, and dimethylbenzene soaks into and paraffin embedding, and section is carried out HE conventional organization and learned dyeing

Observation index and analytical method

Situation is oozed out and after administration in perusal rat wound surface surface, wound surface redness, infection, the newborn epidermal hyperplasia situation of edge of wound, and record wound healing time.After wound, 3d starts to calculate Wound healing rate, adopt transparent template tracing to measure local woanded surface area, calculate by following formula: healing rate=(original wound surface area-existing wound surface area)/original wound surface area × 100%, for healing completely, observe and record each group of healing time with healing rate >=95%.

Experimental result

1. wound healing rate

After wound, 1d naked eyes almost do not observe wound surface variation, therefore go out the data of 1d, in shop, the Wound healing rate of Sanguis Draxonis group, Sanguis Draxonis group and Madecassol group, JINGWANHONG group does not have obvious difference at one time, but all higher than blank group and blank matrix group, difference has statistical significance (P < 0.05), the results are shown in Table 5.

Table 5 trauma in rat different time healing rate comparison (x ± s, n=7)

2. the wound healing time

The wound healing time of Sanguis Draxonis group, Sanguis Draxonis group and Madecassol group, JINGWANHONG group does not have notable difference, but all than blank group and blank matrix group time period, difference has statistical significance (P < 0.05), the results are shown in Table 6.

The impact (x ± s, n=14) of table 6 on the rat wound healing time

Adopt SPSS17.0 to carry out statistical disposition, low dosage Sanguis Draxonis, low dosage Sanguis Draxonis and two kinds of positive drug comparisons, P > 0.05, does not have significant difference, illustrate that low dosage Sanguis Draxonis and Sanguis Draxonis all have the effect that promotes wound healing, Sanguis Draxonis can substitute Sanguis Draxonis simultaneously; Relatively, P < 0.05, has significant difference for low dosage Sanguis Draxonis, Sanguis Draxonis and two kinds of positive drug and blank matrix group, blank group; The Sanguis Draxonis of Sanguis Draxonis, Sanguis Draxonis and the high dose of low dosage, Sanguis Draxonis comparison, P < 0.05, has significant difference, and low dosage effect is better than high dose; Relatively, P > 0.05, does not have significant difference for high dose Sanguis Draxonis, Sanguis Draxonis and blank matrix group, blank group.

The experiment of rabbit wound healing

Experimental agents: Sanguis Draxonis gel preparation, dragon's blood gel preparation and gel-type vehicle: institute of traditional Chinese medicine provides by Kunming, lot number 20130417;

The anti-scar ointment of positive control drug: Mei Jiasi (Madecassol): by Bayer Consumer Care AG, SwitzerI produces, lot number TRTO281; JINGWANHONG RUANGAO: by Tianjin Darentang Wanhong Pharmaceutical Co., Ltd, lot number Z12020440;

Laboratory animal: Japan large ear rabbit, male and female are not limit, and body weight 3～3.5Kg is purchased from the laboratory animal department of the Chinese Academy of Sciences of Kunming medical university, the quality certification number: SCXK (Yunnan) 2011-0004.Raising condition: 25～27 DEG C of temperature, 70 ± 10 DEG C of humidity, the animal drinking-water of freely ingesting.

Divide into groups after 12 rabbit weigh, to be divided at random 2 groups according to body weight, 6 every group.

The local anesthesia of modeling subcutaneous injection lignocaine, 8 wounds of every rabbit, symmetrical, each 4.

First group of 6 rabbit of administration, give respectively low dosage Sanguis Draxonis, low dosage Sanguis Draxonis, blank substrate and Madecassol positive drug; Second group of 6 rabbit, gives respectively high dose Sanguis Draxonis, high dose Sanguis Draxonis, JINGWANHONG positive drug and pure blank.Respectively organize administration every day by said medicine, gauze covers, and medical adhesive tape is wrapped up fixing.Every day wound is retouched to meter simultaneously, take pictures.

Observation index and analytical method

Situation is oozed out and after administration in perusal rabbit wound surface surface, wound surface redness, infection, the newborn epidermal hyperplasia situation of edge of wound, and record wound healing time.After wound, 3d starts to calculate Wound healing rate, adopt transparent template tracing to measure local woanded surface area, calculate by following formula: healing rate=(original wound surface area-existing wound surface area)/original wound surface area × 100%, for healing completely, observe and record each group of healing time with healing rate >=95%.

More than experiment is reached a conclusion, and compound dragon's blood (Sanguis Draxonis) gel and positive control drug and blank wound healing rate aspect have good pharmacology effect compound recipe to answer.Wherein healing rate size is respectively compound dragon's blood (Sanguis Draxonis): Borneolum Syntheticum gel, compound dragon's blood (Sanguis Draxonis): Rhizoma Coptidis: Borneolum Syntheticum gel, lincomycin hydrochloride and lidocaine hydrochloride gel agent, blank group.On incrustation time, decrustation time and healing time, it is also above result.After finding that in test trauma model is built up, a small amount of tissue of mouse skin and Musclar layer is adhered.The skin hardening of wound site after 2 days, no blood flows out, and diet and mental status are normal.After 5 days, wound site starts to have reddish black blood crusts to cover, and after decrustation, wound area obviously diminishes, and wound site is cerise, around a small amount of hair growth of healing skin.After wound heals completely hair also along with wound area reduce increase.After wound heals completely, hair continued growth continues wound coverage position in wound.

Embodiment 2

The preparation of compound dragon's blood (Sanguis Draxonis) gel

After this experiment employing is fully swelling in water by carbomer P940, medicine is joined to solution and fully mix, triethanolamine regulates pH value to make gel reach larger stickiness denseness.Gel should have good glossiness, stretchability, uniformity and centrifugal property.

1. solvent species is investigated

The ratio of determining three kinds of medicines in compound dragon's blood prescription according to document and clinical experience, compound recipe is taking dragon's blood medicine as main.That Sanguis Draxonis has is water insoluble, ether and rare acid solution, is easy to become solution in the physicochemical property of methanol, ethanol and basic solvent.Analyze the relatively dissolving situation of different solvents to compound medicine, select dissolubility good and to the non-irritating solvent of human body the lytic agent as compound dragon's blood medicine.

2. solvent is investigated drug solubility

Precision takes 5 parts, compound dragon's blood medicated powder, and every part of 100mg is dissolved in respectively 5mL glycerol, propylene glycol, Macrogol 200,75% ethanol, 95% ethanol, and fully stirring and evenly mixing, places and observe medicine dissolution situation after 5 minutes, the results are shown in Table 7.

Table 7 different solvents is for the dissolubility (n=0.08) of compound dragon's blood medicine

Remarks: " ++ " dissolves completely, "+" is slightly soluble, "-" be not for substantially dissolving

As can be seen from the above results, compound dragon's blood medicine can dissolve completely in propylene glycol and 75% alcohol solvent.75% ethanol has larger zest to skin, particularly obvious to skin trauma place zest, easily causes wound pain, and open wound patient should not use, and can not use in a large number as drug solvent.Propylene glycol is not only little to human body skin zest but also have Penetration enhancing effect as the lytic agent of compound dragon's blood medicine, make medicine easily see through skin by body absorb enough after performance general action.

3. propylene glycol consumption is investigated

Get every part of 1.55g of equivalent compound dragon's blood medicine, totally six parts.Be dissolved in respectively 5.0mL, 8.0mL, 10.0mL, 15.0mL, 18.0mL, 20.0mL propylene glycol, fully be uniformly mixed, after 15min, observe, when solvent volume is 15.0mL, medicine fully can be dissolved, drug solution is spread upon to nonirritant on skin, better without granular sensation, lubricity.

4. gel-type vehicle is investigated

Can mix homogeneously with medicine for making carrier matrix, compare through 3 kinds of water-soluable gel substrate carbomers, alginate and gelatin, in medicine adition process, the situation such as separate out without layering, caking, medicine using carbomer as substrate and occur, and the gel of preparation becomes that colloidality is good, stable in properties and preparation method simple.Select the carbomer gel solution of accurate concentration, make its better compatibility absorb the drug.After getting compound dragon's blood medicine 1.55g propylene glycol and fully dissolving, join respectively concentration and be 0.5%, 0.6%, abundant stirring and evenly mixing in 0.7%8.45g carbomer gel solution, observe after leaving standstill 2h, the results are shown in Table 8.

Phenomenon (n=2) after table 8 variable concentrations carbomer and medicament mixed

As can be seen from the above results, the carbomer gel substrate that compound dragon's blood medicine is 0.6% with concentration is mixed homogeneously, and has good compatibility at adition process Chinese medicine and substrate, separates out and caking phenomenon without layering, medicine.

5. the investigation of drug loading

The maximum drug loading of ordinary gel agent, generally in 10% left and right, still also can change for its drug loading of difference of ingredient.Gel gross weight is fixed as to 10.0g, investigates the maximum drug loading of substrate to compound dragon's blood medicine.Get respectively compound dragon's blood medicine 0.15g and 0.30g, propylene glycol fully dissolves, and adds after the carbomer substrate of equivalent weight and regulates gel pH value to 7.0 with triethanolamine, finally adds tween 80 to regulate stability and the lubricity of gel, in table 9.

Table 9 drug loading investigation table

Produce the gel of different prescription proportionings according to above prescription substrate, after observing the solute effect of medicine and add carbomer substrate in manufacturing process, whether preparation can there is layering, separate out and caking phenomenon, after adjusting pH value, observes gel outward appearance and mouldability.Experimental result is as follows: compound dragon's blood medicine weight is 0.15g, when the volume of propylene glycol is 2.5mL, outward appearance, the uniformity of gel are better, smear without granular sensation on skin, adding carbomer 9.85g triethanolamine to regulate pH value is 7.0, and gel evenly, occur without in bulk phenomenon.Compound dragon's blood medicine weight is increased to 0.30g, adds carbomer 9.70g, triethanolamine regulates pH to 7.0, and gel is layering immediately, and has block in gel, and granular sensation clearly, is difficult to evenly smear on skin, and stickiness is lower.Compound dragon's blood medicine content is adjusted to respectively to 0.8%, 1.5%, 2.0%, in the time that drug loading is 1.5%, glossiness, the uniformity and the stretchability of preparation all reaches the appearance requirement of gel, 2.0% time, gel medicament contg is large, glossiness obviously reduces, granular sensation is obviously smeared more difficult, the experiment proved that the maximum drug loading 1.5% of compound dragon's blood gel.

6.pH value is investigated

Using the volume of carbomer gel substrate weight, drug weight and propylene glycol as constant basis, it is different pH value that triethanolamine regulates gel, from glossiness, stretchability, the uniformity, four aspects of centrifugal degree, gel outward appearance and stability are carried out to comprehensive grading, select appropriate pH, the results are shown in Table 10.

The comprehensive grading result of the different pH of table 10 compound dragon's blood

Gel is adjusted to different pH value, result is as follows: when pH=6.0, occur obvious lamination, not easy to apply even on skin when pH=6.5, pH=9.0 is irritant for skin wound, pH=7.0 or 8.0 o'clock are without obvious lamination, when pH=8.0 from outward appearance, lamination with all meet gel prescription in the skin situation of smearing.

Embodiment 3

The preparation of compound dragon's blood liniment

The material of Bletilla glucomannan as liniment once selected in this experiment, and it is less for the toxic and side effects of human body that it had both belonged to Chinese herbal medicine, and the Pseudobulbus Bletillae (Rhizoma Bletillae) has the effect of astringing to arrest bleeding, detumescence and promoting granulation, wound repairing.Be widely used in clinically the diseases such as treatment hemoptysis, haematemesis, traumatic hemorrhage, sore swollen toxin, pulmonary tuberculosis hemoptysis, ulcer haemorrhage.In Pseudobulbus Bletillae (Rhizoma Bletillae) tuber, contain abundant Bletilla glucomannan, the jelly that Bletilla glucomannan is a kind of faint yellow homogenizing, water soluble, be insoluble to ethanol, be that one has multiple bioactive Chinese crude drug, be widely used in the production of clinical treatment preparation to there is medical applications widely and be worth.The main preparation methods of Bletilla glucomannan is: (1) decocting bletilla stem piece; (2) will decoct filtering exudate, centrifugal, concentrated; (3) add dehydrated alcohol dehydration, decolouring to make Bletilla glucomannan.The main component of Bletilla glucomannan is neutral Pseudobulbus Bletillae (Rhizoma Bletillae) heteropolysaccharide, belongs to pyranoid form glucomannoglycan, has stronger stickiness.This special nature of the Pseudobulbus Bletillae (Rhizoma Bletillae) both can be brought into play pharmacological action jointly with compound dragon's blood medicine; pharmacological action wound mouth not only to protection astringing to arrest bleeding, wound repairing, detumescence and promoting granulation can also be served as the filmogen of liniment; but in manufacturing process, find that Bletilla glucomannan is comparatively complicated as the liniment manufacturing process of filmogen; and the yield of Bletilla glucomannan is lower, be about 3% left and right.Use Bletilla glucomannan as the filmogen of liniment, after film forming poor, the denseness of toughness lower be not easy on skin fixing, if Bletilla glucomannan concentration is increased, more difficult again when coating, be applied to skin after degree of peeling off poor, be difficult for opening.When after the moisture evaporation in liniment, on skin, be flakey, moistening effect is poor, does not meet the appearance requirement of preparation.So should select other synthetic macromolecule as filmogen.

By being carried out to observation analysis, the single factor of liniment selects suitable filmogen and adjuvant.Polyvinyl alcohol is as main filmogen, film forming speed not only after fast but also film forming effective toughness strong, quality softness.If concentration is too large, stretchability is very poor, difficult coating; Concentration little Yi flows, and can not fix at skin.Need interpolation adjuvant to optimize prescription and improve liniment technique.

Because the drug loading of liniment is less, be suitable for low dose of medicine, the large solvent of dose is difficult for mix homogeneously, and color and the uniformity are all poor, are also in liniment manufacturing process, to need research so select correct drug loading.

1. the investigation of filmogen

Be chosen to membrane material and concentration thereof, polyvinyl alcohol concentration is large, and it is large that the denseness of liniment becomes, and is difficult to smear and extend at the film formation time of skin surface.After the main component of filmogen is determined, add suitable adjuvant and select correct dosage, observe outward appearance and the mouldability of liniment.

Principal element using polyvinyl alcohol weight (A) and carboxyl and sodium carboxymethyl cellulose weight (B) as liniment mouldability, compound dragon's blood medicine weight (C), solvent (D) are respectively propylene glycol and 75% ethanol, as the main formulation factors of liniment mouldability, set up formulation factors table, as following table 11.

The mouldability of table 11 liniment and drug loading test

In sum, select propylene glycol as solvent, 4% polyvinyl alcohol is as the main component of filmogen, and compound dragon's blood medicine weight is 0.20g.

In order to make liniment there is good stretchability, uniformity, mobility, need to add other adjuvant and optimize substrate prescription.

2. liniment substrate formulation optimization and result

The concentration of polyvinyl alcohol is defined as to 4%, and compound dragon's blood medicine weight is 0.2g, propylene glycol 1.0g.Add different adjuvants to carry out Evaluation on Appearance Quality.Matrix formulations is as follows:

(1) concentration is that 1% carbomer 1g joins polyvinyl alcohol as filmogen, adds medicine, and the toughness of gained liniment and degree of peeling off are better, and the uniformity is poor lamination.

(2) concentration is that 5%PVP1g joins in polyvinyl alcohol as filmogen, adds medicine, and the film property of gained liniment is better, and degree of peeling off is better, but to be coated with in membrane process the low viscosity of concentration poor, is difficult for being coated on glass plate.

(3) starch 2g dissolves and joins in polyvinyl alcohol as filmogen, adds medicine, and gained liniment degree of peeling off is better, but gelatinizing phenomenon easily occurs starch in the polyvinyl alcohol that joins dissolving, and the toughness of liniment is poor, in stripping process, easily breaks.

4. polyvinylpyrrolidone 0.5g, sodium carboxymethyl cellulose 0.2g, heating in water bath joins after fully dissolving in the polyvinyl alcohol of dissolving, and liniment film property, degree of peeling off, toughness are better.

3. orthogonal test

According to the result of above experiment, filmogen and adjuvant to liniment further screen, and adopt the method for orthogonal test, filter out and affect the principal element of film forming and the suitableeest weight.Investigate 4 principal elements that affect mouldability: the weight (D) of weight (A), sodium carboxymethyl cellulose (B), polyvinylpyrrolidone (C) and the medicine of polyvinyl alcohol is examined or check, 3 levels of each selecting factors, press L9 (34) and arrange orthogonal test, factor and level are shown according to above experiment conclusion and set out and set up horizontal factor and orthogonal test table, respectively in table 12 and table 13.

Table 12 factor level table

Table 13 orthogonal test table

Above nine groups of orthogonal test conclusions are as follows: the content of polyvinyl alcohol is high, and liniment stickiness is larger, and very difficult painting is opened and film formation time on skin extends.Sodium carboxymethyl cellulose has the effect that regulates membrane mobility, and dosage increases makes the denseness of liniment strengthen, and plasticity strengthens, but toughness drop.The drug loading of liniment is less, if dose increases, made liniment is inhomogeneous, and color, the uniformity and mouldability are poor, and outward appearance does not meet liniment requirement.By after the moisture evaporation in liniment, obtained liniment peels off on skin that difficulty, fragility are large, poor toughness, nonelastic, be not easy to stretch, dullish.Add glycerol in manufacturing process and increase pliability and the glossiness of liniment as plasticizer.

4. liniment scoring criterion

Stretchability: liniment is easily coated with and opens on skin, and preparation exquisiteness, without granular sensation.2.5 are divided into full marks.

Uniformity: liniment is without lamination, uniform particles, 2.5 are divided into full marks.

Pliability: on skin after solvent evaporates, easy fracture not in stripping process, flexible and elasticity, 2.5 are divided into full marks.

Mobility: mobility can not be too strong, and semisolid preparation, is divided into full marks with 2.5.

5. compound dragon's blood liniment formulation and technology is optimized orthogonal test

Investigate according to above result of the test, filmogen composition is respectively polyvinyl alcohol, polyvinylpyrrolidone and sodium carboxymethyl cellulose, and solvent is propylene glycol, and plasticizer is glycerol.Investigate result according to above test, stretchability, uniformity, pliability and the mobility tool of the each composition in substrate to compound dragon's blood liniment has a certain impact.Be wherein the dosage of polyvinyl alcohol, polyvinylpyrrolidone and glycerol on the larger factor of mouldability impact of liniment, the variation relative complex between each factor.In order further to optimize and filter out the best prescription of liniment, adopt Orthogonal Experiment and Design further to optimize preparation technology, using the comprehensive grading of presentation quality and mouldability as investigating index, investigate 4 principal elements that affect mouldability: polyvinyl alcohol weight (A), polyvinylpyrrolidone weight (B), the weight (C) of medicine and amounts of glycerol (D), 2 levels of each selecting factors, press L8 (24) and arrange orthogonal test, set out factor level table in table 14 according to above experiment conclusion, orthogonal experiments is in table 15, variance analysis is in table 16.Wherein in above prescription, sodium carboxymethyl cellulose is 0.2g, propylene glycol 1.0g, and the concentration of polyvinyl alcohol is 4% in each prescription, to be fixed amount.

Table 14 factor level table

Table 15 Orthogonal experiment results

Table 16 the results of analysis of variance

From table 14 result, using liniment outward appearance mouldability and cutaneous sense as index, by intuitive analysis, because the weight factor extreme difference of polyvinyl alcohol is larger, so the weight of polyvinyl alcohol is larger to the quality influence of liniment, each factor is followed successively by A>C>D>B to the influence degree of liniment outward appearance mouldability, is respectively weight, drug weight, amounts of glycerol and the polyvinylpyrrolidone weight of polyvinyl alcohol.Best of breed is A1B3C3.

The result of These parameters is carried out to variance analysis, the result that outward appearance mouldability is affected to difference by the each factor of table 14 can be found out, the weight of polyvinyl alcohol is on significantly (P<0.05) of the impact of liniment, compare and can find out that dosage mouldability impact for liniment in four factors of polyvinyl alcohol is larger by F, weight to polyvinyl alcohol in manufacturing process pays particular attention to, consider the impact of each factor, amounts of glycerol and polyvinylpyrrolidone are not remarkable on mouldability impact, therefore selecting the polyvinyl alcohol weight of 4% concentration is 8.0g, medication amount is 0.2g, amounts of glycerol is 0.5g, polyvinylpyrrolidone is 0.5g.Optimized process flow is A1B1C2D1.

6. prescription confirmatory experiment

According to orthogonal experiments analysis, determine that optimizing prescription is: 4% polyvinyl alcohol is 8.0g

Propylene glycol is that 1.0g sodium carboxymethyl cellulose is 0.2g

Take medicine by experiment recipe quantity, repeatedly add on a small quantity, being dissolved in propylene glycol 1.0g makes it fully dissolve the phase as A, in polyvinyl alcohol after dissolving, take by prescription weight, add sodium carboxymethyl cellulose 0.2g and polyvinylpyrrolidone 0.5g, 80 DEG C of left and right of water-bath make the abundant swelling dissolving of adjuvant as B phase, and B is added to the A mixing that stirs in mutually, finally add surfactant tween 80 to keep the stability of liniment.Liniment is evenly spread upon on the glass plate that scribbles liquid paraffin, 37 DEG C of oven dry in baking oven, observe outward appearance uniformity, stretchability, mobility and the film pliability of liniment and carry out comprehensive grading.

7. demonstration test and scoring

In order to verify the accuracy of the above results, to guarantee liniment preparation technology's reasonability, by above-mentioned process conditions, arrange to repeat experiment, result of the test is in table 17.

The orthogonal confirmatory experiment result of table 17

Reach a conclusion as follows by the demonstration test of above liniment: the liniment of orthogonal test gained is than in experiment, preferably out liniment mobility is slightly poor in character, and denseness is larger, while smearing, be difficult for being evenly coated with and opening, on skin, smear preparation thickness and differ.May be because the consumption of polyvinylpyrrolidone reduces, the film property of polyvinyl alcohol is not adjusted to optimal value, so still use prescription substrate that comprehensive grading is the highest as final process route for this kind of phenomenon.In coating process, need to reduce the generation of bubble as far as possible, slide is volatilized and dried 37 DEG C of left and right, approach under human body temperature and volatilize, accelerate film forming speed.

Embodiment 4

The preparation of compound dragon's blood cataplasma

Catablasm base material mainly contains the compositions such as adhesive agent, wetting agent, cross-linking agent and viscosifier, and water content, adhesion, bioavailability, comfort and the breathability of substrate to cataplasma plays mastery reaction.

1. adhesive agent is selected

This experiment is used respectively hydrophilic compounds sodium polyacrylate and polyvinyl alcohol separately as the framework material of cataplasma, and after result adds cross-linking agent and wetting agent in sodium polyacrylate, cohesiveness is very large, and substrate is easily agglomerating, coating difficulty.Use separately polyvinyl alcohol as framework material, the easy membranization of stromal surface, viscosity is very little, because substrate cohesiveness reduces whole substrate denseness compared with little, almost can not be coated with.Framework material using the mixture of polyvinyl alcohol and sodium polyacrylate as cataplasma repeatedly still has good adhesion, and can delay the gelation rate of sodium polyacrylate after skin adhesive, crosslinking time extends, cohesiveness suitably reduces, and the coating time is extended relatively, is conducive to coating.

2. wetting agent is selected

Wetting agent has been selected respectively glycerol and two kinds of solvents of propylene glycol, and compare propylene glycol through experiment and as wetting agent, the cohesiveness of mastic is significantly reduced, insufficient formability, and also poor compared with glycerol in gloss appearance sense and pliability.Glycerol can account for the 30%-50% of whole mastic weight as wetting agent, through test of many times comparison, amounts of glycerol account for total mastic weight 45% time, not only there is good glossiness but also cohesiveness better, mouldability is better.

3. cross-linking agent is selected

Cross-linking agent is selected tartaric acid and aluminium hydroxide, and soda acid combined effect is in carrier matrix, promotes the abundant mixing of framework material and adhesive and crosslinked, and crosslinking time is rapid.

4. tackifier is selected

With sodium polyacrylate as main skeleton filmogen, select respectively two groups of auxiliary material as catablasm base material of polyvinylpyrrolidone and sodium carboxymethyl cellulose, it is serious in any viscous force that experimental result shows that polyvinylpyrrolidone experimental group oozes cloth, the degree of peeling off of sodium carboxymethyl cellulose experimental group better and nothing ooze cloth phenomenon.Sodium carboxymethyl cellulose is suitable as the tackifier in this prescription.

5. cataplasma orthogonal test

Catablasm base material list factor result is observed, to ooze cloth, the overall merit of skin tracing ability and mouldability is as investigating index, 7 factors that investigation affects mouldability are respectively: sodium polyacrylate (A), polyvinyl alcohol (B), aluminium hydroxide (C) and tartaric acid (D), sodium carboxymethyl cellulose (E), drug weight (F), the ratio (G) of propylene glycol and medicine, 3 levels of each selecting factors, press L18 (37) and arrange orthogonal test, design factor level in table 18 according to above experiment conclusion, Orthogonal Experiment and Design is in table 19.

Table 18 factor level table

Table 19 orthogonal test table

Carry out the making of cataplasma technical study according to above prescription substrate composition, subject matter and conclusion are as follows: 1) ooze cloth more obvious 2) cataplasma is short standing time, may fail full cross-linkedly due to medicine and substrate, and on skin, take off to paste and while peeling off, easily fall medicine.3) the amount difference of cross-linking agent, crosslinking time is also different, and the amount of cross-linking agent is larger, and the crosslinking rate of mastic is fast, and mastic is easily solidifying.4) add tartaric acid should be and repeatedly add on a small quantity in mastic, fully mix homogeneously with mastic, cross-linking effect is good.

After above cataplasma is placed to a week, carry out overall merit through cutaneous sense test and presentation quality, eliminate for the cataplasma prescription that has obvious problem, wherein significantly problem is: ooze cloth serious, without any viscous force; The initial bonding strength of substrate is poor, after skin is pasted repeatedly, easily comes off; Initial bonding strength > cohesiveness is easy to depart from non-woven fabrics at stripping process mesostroma.Presentation quality: serious owing to oozing cloth, mastic fails to keep moisture, mastic is without any viscous force, and mastic mouldability is poor; Pasty consistence is low, can make mastic easily flow out when coating, and embossability is poor.

6. catablasm base material ratio optimization experiment

Observe through many experiments, in document, Zhang Lei waits for cross-linking agent and selects and forge with the time to armful arriving, and cross-linking agent ratio and crosslinking time are for mouldability, the skin tracing ability of cataplasma and can have a certain impact by the equal tool of coating.With film residual, cohesiveness, hold viscous force and degree of peeling off carries out overall merit, investigate 4 principal elements that affect mouldability: propylene glycol (A), sodium carboxymethyl cellulose (B), aluminium hydroxide and tartaric ratio (C), crosslinking time (D) are examined or check, 2 levels of each selecting factors, arrange orthogonal test by L8 (24).Factor level is in table 20, and orthogonal test is in table 21.

Table 20 factor level table

Table 21 orthogonal test table

Make cataplasma according to above prescription substrate composition, result aluminium hydroxide: the crosslinking rate speed than 2:1 when tartaric acid=1:1 is fast, crosslinking time shortens, and mastic is easily solidifying.Crosslinking time should determine in 10～20min, colloid condense in bulk when the time exceedes after 25min mastic coating, and more difficult painting is opened.Above cataplasma is carried out to overall merit, and from oozing cloth situation, mastic residual, hold viscous force and degree of peeling off carries out integration test, measuring operating weight while holding viscous force is 500g counterweight, and degree of peeling off is 100g counterweight.Cataplasma area is 3cm × 3cm, holds viscous force method of testing: cataplasma is attached to smooth rustless steel iron plate upper, hangs 500g counterweight below, record cataplasma and come off the time.Degree of peeling off method of testing: cataplasma is attached to smooth rustless steel iron plate upper, clamps with clip on the non-woven fabrics apart from mastic 2cm, counterweight is positioned over below clip, mastic face is become to the angle of 180 ° with rustless steel iron plate face.Record the time that cataplasma comes off completely from corrosion resistant plate.

7. comprehensive evaluation result

No. 1: ooze on a small quantity cloth, cohesiveness is better, repeatedly take off subsides, residual without substrate, fall on a small quantity medicine.Hold viscous force 46s, splitting time 12s, while peeling off, a large amount of substrate comes off.

No. 2: it is comparatively obvious to ooze cloth situation, and cohesiveness is better, and initial bonding strength is larger, without falling medicine situation.Hold viscous force 115s, splitting time 234s.

No. 3: have the cloth of oozing situation, compared with No. 2 lighter, cohesiveness is better, initial bonding strength is compared with a little less than 1,2.Hold viscous force 133s, splitting time 355s.

No. 4: have the cloth of oozing, similar to No. 3, cohesiveness is better, stickiness is little compared with 1, No. 2, falls medicine situation not too obvious.Hold viscous force 148s, splitting time 144s, while peeling off, a large amount of substrate comes off.

No. 5: it is similar to No. 1 to ooze cloth situation, and cohesiveness is better, and stickiness is similar to 1, No. 2.Fall on a small quantity medicine.Hold viscous force 49s, degree of peeling off 128s, while peeling off, a large amount of substrate comes off.

No. 6: ooze on a small quantity cloth, initial bonding strength is larger, and cohesiveness is better, fall on a small quantity medicine.Hold viscous force 88s, splitting time 83s.

No. 7: several nothings are oozed cloth, and cohesiveness is fine, initial bonding strength, compared with a little less than 1, No. 2, falls medicine on a small quantity.Hold viscous force 172s, splitting time 65s, a small amount of substrate comes off.

No. 8: ooze on a small quantity cloth, cohesiveness is fine, initial bonding strength is similar to No. 5, falls on a small quantity medicine.Hold viscous force 59s, splitting time 110s, a small amount of substrate comes off.

In sum, through the adjustment of repetition test repeatedly and prescription matrix components ratio, the cloth phenomenon of oozing of cataplasma is obviously improved, good forming ability, mastic initial bonding strength and hold viscous force and increase.

8. optimize cataplasma preparation technology

For cataplasma outward appearance and quality standard are met to state-promulgated pharmacopoeia regulation, be optimized again for above cataplasma formulation and technology, main direction is as follows:

1) adjuvant of selecting is more few better, so not only makes cost, and can avoid making because of increasing of adjuvant the drug loading of cataplasma to reduce.

2) simplify processing step, make cataplasma in commercial production journey, reduce some unnecessary intermediate links, reduce the waste of manpower and financial resources.

3) presentation quality and instrument test index meet pharmacopeia regulation, and human body skin is had to good affinity, and skin tracing ability is better.

9. framework material is preferred

Sodium polyacrylate has good stickiness and mouldability compared with other framework material, the proportional relation of dosage of first viscous force and use, good forming effect and easy to control in following experiment still as framework material.

10. Optimization of Adjuvant

1) relatively find through experiment, cross-linking agent uses dihydroxyaluminum aminoacetate crosslinking time than the relative prolongation of aluminium hydroxide, crosslinking time length can not only make cross-linking agent fully mix with framework material, and has the more time to use coating machine to be coated with, and is more suitable for the commercial production of cataplasma.

2) wetting agent is still selected glycerol, because medicine can not dissolve completely in glycerol, so will add a certain amount of glycerol after medicine dissolution with appropriate propylene glycol, not only can keep cataplasma surface gloss and wettability, and glycerol has a good moisture retention to human body is without any side effects, after repeatedly pasting, cataplasma presentation quality is without significant change.

3) adjuvant addition sequence and forge the mouldability on cataplasma and mastic stickiness with the time and have appreciable impact, answers selecting properly adjuvant and determines that according to mutual relation between adjuvant and framework material and effect feature adjuvant addition sequence and control forges and the time.

11. cataplasma optimization of orthogonal test experiments

Blank substrate matched group and medicine group cataplasma compare, and ooze cloth, cohesiveness, uniformity, film residual quantity, initial bonding strength, hold the aspect such as viscous force and degree of peeling off test and carry out overall merit from mastic.Redefine cataplasma prescription composition and ratio.Consider to affect 4 principal elements of mouldability: sodium polyacrylate (A), dihydroxyaluminum aminoacetate (B), tartaric acid (C), polyvinylpolypyrrolidone (D) are examined or check, 3 levels of each selecting factors, press L9 (34) and arrange orthogonal test, factor level is in table 22, and orthogonal test is in table 23.

Table 22 factor level table

Table 23 orthogonal test table

12. cataplasma presentation qualities and cutaneous sense test synthesis are evaluated

No. 1: ooze cloth serious, cohesiveness > sticks base power, and initial bonding strength is very large.

No. 2: ooze on a small quantity cloth, mastic mouldability is poor, and initial bonding strength is very large, and cohesiveness is less, in mastic, contain a large amount of bubbles and bubble cavity.

No. 3: several nothings are oozed cloth, in stripping process, a small amount of substrate sticks to cover and is lining with, and initial bonding strength is larger, and cohesiveness is less, contains a large amount of bubbles and a small amount of bubble cavity in mastic, and glossiness is better.

No. 4: ooze on a small quantity cloth, severally in stripping process stick to lid without substrate and be lining with, mastic uniformity is better, and mouldability is better, and cohesiveness > initial bonding strength contains a large amount of bubbles in mastic, and glossiness is fine, fine and smooth sense.

No. 5: several nothings are oozed cloth, in stripping process, almost stick to lid without substrate and be lining with, mastic uniformity is better, and mouldability is better, and cohesiveness > sticks base power, and initial bonding strength is less than No. 4, and bubbles volume is a lot, and glossiness is good.

No. 6: ooze on a small quantity cloth, severally in stripping process stick to lid without substrate and be lining with, mastic uniformity is better, cohesiveness > initial bonding strength, bubble is more, and glossiness is better, rich fine and smooth sense.

No. 7: several nothings are oozed cloth, in stripping process, be lining with at lid without matrix adhesion, mastic uniformity is fine, and mouldability is fine, initial bonding strength > cohesiveness, bubble is more, and glossiness is better.

No. 8: several nothings are oozed cloth, in stripping process, be lining with at lid without matrix adhesion, mastic uniformity is fine, and mouldability is fine, cohesiveness > initial bonding strength, bubble is more, and glossiness is good.

No. 9: several nothings are oozed cloth, in stripping process, be lining with at lid without matrix adhesion, cohesiveness > initial bonding strength, mastic uniformity is better, good moldability, No. <7, initial bonding strength, bubbles volume is large, and glossiness is good.

13. initial bonding strengths, hold viscous force and degree of peeling off and measure

Cataplasma is made into fixed-area 7.5cm × 11cm, every cataplasma is carried out first viscous force, held viscous force and degree of peeling off mensuration by tester respectively.

14. initial bonding strength tests

Initial bonding strength will be apart from being made as 10cm, and bead can not be by the distance of 10cm for tangling, using its ball as first viscous force test result in one minute.

15. hold viscous force test

Hold viscous force method of testing: the cataplasma of 7.5cm × 11cm is attached on corrosion resistant plate, for 2kg rubber wheel rolls 3 times repeatedly, hangs 500g counterweight below steel plate with heavily, measure the time that cataplasma is come off completely by corrosion resistant plate.

16. degree of peeling off tests

The assay method of degree of peeling off: the cataplasma of 2.5cm × 10cm is attached on corrosion resistant plate with heavily taking turns and repeatedly roll 3 times for 2kg rubber, steel disc is fixed on and is peeled off on tester, it is 200N that pulling force is peeled off in setting, speed is 150m/min, and peeling off minute is that cataplasma is by the time of peeling off completely on corrosion resistant plate.

Cataplasma to difference prescription proportioning carries out instrument test, because the cataplasma presentation quality of front 3 prescription proportionings is poor undesirable, all the other cataplasmas are carried out respectively just viscous force (A), hold viscous force (B), degree of peeling off (C) test, and test result is in table 24.

Table 24 just viscous force, hold viscous force, degree of peeling off test result

By above cataplasma integration test comparison, not only presentation quality is better for No. 8 cataplasma prescription substrate, and the test result of each power is relatively high.Just viscous force is larger for skin test entirety sensation, and cohesiveness reduces relatively.

17. prescription viscosifier regulation experiment groups

On this experiment basis, change the ratio composition of viscosifier and cross-linking agent, continue the experiment of screening catablasm base material prescription, carry out overall merit from presentation quality, adhesion strength, glossiness and the test of each power of cataplasma.Adjust the ratio of adhesive agent, wetting agent and cross-linking agent.Sodium polyacrylate (A), sodium carboxymethyl cellulose (B), dihydroxyaluminum aminoacetate (C), tartaric acid (D), glycerol (E) and six factors of pH (F) are investigated to experiment arrangement.Experimental group table is in table 25.

Table 25 viscosifier regulation experiment

18. presentation qualities and cutaneous sense test evaluation result

No. 1: trace oozes cloth, there is a small amount of bubble viscous force less, in stripping process, remain on backing without substrate.

No. 2: several nothings are oozed cloth, and reflecting feel is poor, and initial bonding strength is better, there are a large amount of bubbles, it is residual that in stripping process, there is a small amount of substrate at backing edge.

No. 3: several nothings are oozed cloth, and bubble is less, and initial bonding strength is larger, in stripping process, remain on backing without substrate, but the fine and smooth sense of glossiness is poor.

No. 4: ooze cloth obvious, bubble is more, in stripping process, remain on backing without substrate, glossiness and fine and smooth sense are better.

No. 5: ooze cloth obvious, bubble is more, in stripping process, remain on backing without substrate, glossiness and fine and smooth sense are better.

No. 6: ooze on a small quantity cloth, bubble is more, in glass process, remain on backing without substrate, the fine and smooth sense of glossiness is better.

No. 7: trace oozes cloth, a large amount of bubbles, the fine and smooth sense of gloss is better, residual without substrate in stripping process.

No. 8: ooze on a small quantity cloth, more bubble, initial bonding strength is better, and glossiness, fine and smooth sense are better, residual without substrate in stripping process.

No. 9: ooze on a small quantity cloth, bubble is less, glossiness, fine and smooth sense are better, residual without substrate in stripping process.

No. 10: several nothings are oozed cloth, and bubble is less, and initial bonding strength is less, residual without substrate in stripping process.

The use amount of cross-linking agent is different from ratio, and in manufacturing process, the viscosity of mastic is also different, and larger its denseness of ratio is less, so select the impact that suitable ratio is larger on forming of cataplasma with amount.

19. initial bonding strengths, hold viscous force and degree of peeling off test result

Cataplasma is carried out respectively just viscous force (A), holds viscous force (B), degree of peeling off (C) test, and instrument test the results are shown in Table 26.

Table 26 just viscous force, hold viscous force, degree of peeling off test result

In mensuration process, the viscous force of sodium carboxymethyl cellulose is significantly less than polyvinylpolypyrrolidone, in mensuration degree of peeling off, is easy to peel off, but because degree of peeling off is very little, countless certificates in mensuration process, have to a certain degree so measure the big or small viscous force of degree of peeling off.Hold viscous force and be starkly lower than viscosifier polyvinylpolypyrrolidone group, but just viscous force is moderate in cutaneous sense test, increase mutually without sticky chaeta phenomenon cohesiveness taking off subsides process.

In this experiment, write out a prescription as the final formulation and technology of cataplasma using this, be also optimized on this basis later, produce up-to-standard cataplasma product.

Embodiment 5

Compound dragon's blood (Sanguis Draxonis) preparation [Sanguis Draxonis: Borneolum Syntheticum 9:1 is to Rabbits with Acute toxicity, skin irritation and anaphylaxis experiment

Laboratory animal: healthy adult Japan large ear rabbit, male and female are not limit, body weight 2.5～3.5Kg; Healthy adult Cavia porcellus, male and female are not limit, body weight 250～450g; By Kunming medical university Experimental Animal Center provide [quality certification SCXK(Yunnan) 2011-0004.25 DEG C ± 3 DEG C of ambient temperatures, relative humidity 70% ± 15%, light application time 12h/12h.

Skin acute toxicity testing

Select 20 Japan large ear rabbits to be divided at random 6 groups, 5 every group.Be intact skin gel-type vehicle matched group, intact skin compound dragon's blood gel group, intact skin compound dragon's blood gel group, damaged skin gel-type vehicle group, damaged skin compound dragon's blood gel group, damaged skin compound dragon's blood gel group.Before experiment, hair is taken off with razor, depilation scope be 150cm2(approximately quite rabbit body surface area 10%), after 24h, check whether shave hair-fields has damage, reject skin damage rabbit, subsequently tested medicine is evenly applied to depilation district, and is fixed sub-cage rearing with non-stimulated gauze; Damaged skin group first will be lost hair or feathers position with after 75% alcohol disinfecting, cause local scratch with emery cloth paper in depilation position friction, and taking oozing of blood as degree, immediately in damaged part medication, concrete dosage and coating method are with above-mentioned intact skin group.After 24h, take off binder, with after warm water cleaning tested material, observe and record the performance of rabbit whole body and poisoning situation every day, specifically comprise the variation of the variation, breathing, circulation, central nervous system, extremity activity etc. of body weight, depilation district red swelling of the skin degree, hair, eye and the mucosa of animal, if chance animal dead, postmortem in time, perusal is also carried out histopathologic examination.

Skin irritation test

Single-dose skin irritation test is got 20 of healthy Japan large ear rabbits, and male and female are not limit, and is divided at random 4 groups, i.e. intact skin and damaged skin compound dragon's blood gel and compound dragon's blood gel group, 5 every group.Before experiment, hair is taken off with razor, depilation scope be 150cm2(approximately quite rabbit body surface area 10%).After depilation, intact skin group by tested material primary coating on the skin of animal subject; Damaged skin group first will be lost hair or feathers position with after 75% alcohol disinfecting, cause local scratch in depilation position friction with dry sanding paper, taking oozing of blood as degree, immediately in damaged part administration, and fixed with nonirritant gauze, after 24h, use warm water cleaning medicine, observation 1,24,48 and 72h medicine-feeding part have or not the situation such as erythema, edema, and result is marked and does stimulus intensity evaluation according to document skin irritation reaction standards of grading.

Multiple dosing skin irritation test is got 20 of Japan large ear rabbits, grouping and making damaged skin are the same, successive administration 7d, after each coating, fix 12h with hospital gauze and adhesive tape, after last administration 24h, clean medicine with warm water, observe 1,24,48 and 72h medicine-feeding part have or not the situation such as erythema, edema, result is marked and does stimulus intensity evaluation according to document skin irritation reaction standards of grading.Carry out dermoreaction scoring by table 27, carry out overall merit with the meansigma methods of animal subject integration, according to 24,48 and 72h respectively observe time point top average, judge skin irritation intensity by table 28, the results are shown in Table 29.

Experiment conclusion: Sanguis Draxonis group, Sanguis Draxonis group, blank machine-processed group carry out rabbit single-dose skin irritation test and show, three is all without any zest, use that can safety.

Table 27 skin irritation reaction scoring

Table 28 skin irritation strength grading

The each group of the table 29 multiple dosing skin irritation table of integrals

Skin hypersensitivity experiment

Healthy adult Cavia porcellus, male and female are not limit, and are divided at random 5 groups, blank group, gel-type vehicle group, compound dragon's blood gel group, compound dragon's blood gel group and 2,4-dinitrochlorobenzene group (being configured to before use the concentration that excites of 1% sensitization concentration and 0.1%), 10 every group.Before experiment, with razor, hair is shaved totally, scope 3cm × 3cm, checks after 24h whether shank-feathering district has damage, rejects skin damage Cavia porcellus.

Sensitization contact is by gel-type vehicle, compound dragon's blood gel, compound dragon's blood gel (every 0.2g) and 2,4-dinitrochlorobenzene (every 0.2ml) is evenly applied in depilation district, right side, the back skin of each group of corresponding Cavia porcellus after respectively at depilation after 24h, cover with gauze, fixing with non-stimulated adhesive plaster wrapping, sub-cage rearing, removes left drug with warm water after lasting 6h.7d and 14d, in kind repeat once.

Excite the 14d being contacted with after last administration sensitization, by gel-type vehicle, compound dragon's blood gel, compound dragon's blood gel and 2,4-dinitrochlorobenzene (concentration 0.1%, dosage 0.2ml) be evenly applied in depilation district, left side, the back skin of each group of corresponding Cavia porcellus, cover with gauze, fixing with non-stimulated adhesive plaster wrapping, sub-cage rearing, removes left drug with warm water after lasting 6h.At once (0h) observes dermoreaction situation, then in 24,48 and 72h continue to observe skin allergy situation.Result is marked and sensitization evaluation by document skin allergy standards of grading, and by calculate Cavia porcellus sensitivity response meansigma methods with formula: sensitivity response meansigma methods=(erythema forms total score+edema and forms total score)/animal number of cases; Number of animals/animal subject sum * 100% of sensitization incidence rate=occur erythema or edema (no matter degree weight).The anaphylactoid scoring of evaluating skin can, by the standard of table 30, be evaluated by table 31.

Table 30 skin allergy degree standards of grading

Table 31 hypersensitive evaluation criterion

Data statistic analysis: the data of all experimental results are with mean ± standard deviation (x ± s) represent, all data analysiss carry out on SPSS17.0 statistical package, carry out the comparison of many group differences with one factor analysis of variance, when P < 0.05, think that difference has statistical significance.

6. sign inspection

Compound dragon's blood gel, compound dragon's blood gel, gel-type vehicle are and cause anomalous effects complete, skin conditions, feed, stool, breathing, heart beating, eye and the mucosa of damaged skin rabbit, central nervous system, extremity activity; Carry out the body weight difference of 3d, 7d, 14d before coating, after coating, we find compound dragon's blood gel group, compound dragon's blood gel and the comparison of gel-type vehicle group matched group, there was no significant difference (table 32).There is not any acute toxic reaction in 6 groups of rabbit in observation period 14d, dead without 1 example.Experimental result shows, compound dragon's blood gel, compound dragon's blood gel to intact skin and damaged skin skin all without obvious acute toxic reaction.

The anxious malicious the weight of animals statistical result of table 32 rabbit percutaneous dosing

8. skin appearance and morphological observation

In experiment, perusal find, Sanguis Draxonis gel, dragon's blood gel to normal skin there are no any harmful effect; To damaged skin, gel-type vehicle group 3d before and after administration, without significant change, starts incrustation after 4d, and Sanguis Draxonis gel and dragon's blood gel 1d after coating starts incrustation, and after 4d, incrustation starts to come off.Experiment show Sanguis Draxonis gel and dragon's blood gel to rabbit normal skin without any harmful effect, damaged skin is had to the effect of wound healing.

9. single-dose skin irritation test

After single is smeared Sanguis Draxonis gel, dragon's blood gel and gel-type vehicle, rabbit intact skin has no the irritant reaction such as erythema or edema, except damaged skin gel-type vehicle matched group is after administration 2d incrustation, and since 4d, the decrustation of all healing of damaged wound.

10. multiple dosing skin irritation test

After repeatedly smearing Sanguis Draxonis gel, dragon's blood gel and gel-type vehicle, rabbit intact skin has no the irritant reaction such as erythema or edema, damaged skin group Sanguis Draxonis gel, dragon's blood gel are since 3d, damaged wound healing decrustation, again cause oozing of blood with dry sanding paper, administration, wound healing after 7d, returns to normal condition.

11. skin hypersensitivity experiments

As shown in Table 33, DNFB group coating position has 10 Cavia porcelluss to occur obvious erythema at 0h, all occurs erythema at 24h and 48h; Gel-type vehicle, Sanguis Draxonis gel group and dragon's blood gel treated animal coating position are without erythema edema phenomenon, also without systemic anaphylaxis phenomenons such as asthma, astasia or shocks.

Table 33 Sanguis Draxonis gel, the anaphylaxis impact of dragon's blood gel on Cavia porcellus

Embodiment 6

Compound dragon's blood (Sanguis Draxonis) preparation [Sanguis Draxonis: Borneolum Syntheticum 9:1 rabbit long term toxicity test.

Laboratory animal healthy adult Japan large ear rabbit, male and female are not limit, and body weight 2.5～3.5Kg provides [quality certification SCXK(Yunnan) 2011-0004] by Kunming medical university Experimental Animal Center.25 DEG C ± 3 DEG C of ambient temperatures, relative humidity 70% ± 15%, light application time 12h/12h.

Animal divides into groups according to the requirement of " study of tcm new drug guide (pharmacy pharmacology's toxicology) " and " herbal pharmacology research methodology ", gets 20 rabbit, is divided at random 4 groups, 5 every group according to body weight.The blank matrix group of intact skin, compound dragon's blood group, compound dragon's blood group, the blank matrix group of damaged skin, compound dragon's blood group, compound dragon's blood group.

The maximum dosage of medication, every day 1 time, continuous 2 weeks.Coating position is a depilation district, rabbit back spinal column both sides of family (with razor, hair is shaved off, depilation scope is 150cm2, approximately quite 10% of rabbit body surface area), is fixed sub-cage rearing with non-stimulated gauze; Damaged skin group first will be lost hair or feathers position with after 75% alcohol disinfecting, cause local scratch with emery cloth paper in depilation position friction, and taking oozing of blood as degree, immediately in damaged part medication, concrete dosage and coating method are with above-mentioned intact skin group.

Observation index

Ordinary circumstance observe: during administration and observe and record every day convalescent period rabbit whole body show and poisoning situation, specifically comprise the variation of the variation, breathing, circulation, central nervous system, extremity activity etc. of body weight, depilation district red swelling of the skin degree, hair, eye and the mucosa of animal, if chance animal dead, postmortem in time, perusal is also carried out histopathologic examination.

Hematology detects: administration 2 weeks, in the time of drug withdrawal next day and drug withdrawal two weeks (convalescent period), detect, and get 2 rabbit for every group.Before detecting, fasting be can't help water approximately 12 hours, and fixing animal is adopted whole blood in the fixing Rabbit central ear artery of regaining consciousness with disposable blood taking tube.Detection index comprises: erythrocyte (RBC) counting, hemoglobin (Hb), red cell volume (RDW), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) (MCHC), reticulocyte (RET%) counting, leukocyte (WBC) and classification (NE%, LY%, MO%), platelet (PLT) counting, prothrombin time (PT), Fibrinogen (FIB); And bone marrow smear inspection.

Blood biochemical detects: first the 1st week of administration, administration 4 weeks and recovery finish to detect in 24h for 2 weeks.Before detecting, animal needs fasting to can't help water approximately 12 hours, adopt whole blood 3～6.5ml in fixing clear-headed Rabbit central ear artery with disposable blood taking tube, under room temperature, leave standstill approximately 20 minutes, with the centrifugal 10min of 3000r/min, take out serum and carry out aspartic transaminase (AST), alanine aminotransferase (ALT), alkali phosphatase (ALP), gamma glutamyltransferase (γ-GGT), blood urea nitrogen (Bun), creatinine (Cre), total protein (TP), albumin (ALB), blood glucose (GLU), total bilirubin (TBIL), creatine phosphokinase (CK), T-CHOL (TC), triglyceride (TG), sodium ion, potassium ion, chloride ion, calcium ion concentration and pH value.

System is dissected and histopathological examination administration finishes for 4 weeks, after indices detects, randomly draw 2 animals and dissect for every group, residue animal carries out restorative observation, observation end at indices detect after fasting can't help the about 12h of water anaesthetize put to death dissection, carry out the histopathological examination of medicine-feeding part skin and each vitals, calculate organ coefficient.

The data of all experimental results of data statistic analysis are with mean ± standard deviation (x ± s) represent, all data analysiss carry out on SPSS17.0 statistical package, carry out the comparison of many group differences with one factor analysis of variance, with the relatively difference of mean between two groups of Dunnett inspection; When P < 0.05, think that difference has statistical significance.

Embodiment 7

The explanation method of quality control as an example of compound dragon's blood (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1) example

Instrument is worn peace P680 high performance liquid chromatogram instrument; Wear peace chromatographic work station; Wear peace UVD170 variable-wavelenght detector; Thermo BDS HYPERSIL C18(250mm*4.6mm, 5 μ m)

Reagent and reagent 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, lourerin B.Purchased from national drug biological products assay institute; Methanol, acetonitrile is chromatographically pure, and water is the rear heartily pure water of filter, and other reagent are analytical pure; Compound dragon's blood gel (self-control, lot number 130516,130517)

Resina Draconis-drug alcohol-insoluble substances assay

Take Resina Draconis-drug 2g and be wrapped in filter paper, Soxhlet is extracted into colourless, volatilizes filter paper, and alcohol-insoluble substances content is 0.297%, is no more than the alcohol-insoluble substances that pharmacopeia specifies and is no more than 1.5%.

Sanguis Draxonis trap checks

Instrument: TU-1810 ultraviolet-uisible spectrophotometer (Beijing Puxi General Instrument Co., Ltd)

Take 4g Sanguis Draxonis in 10ml volumetric flask, add ultrasonic the making of methanol to dissolve, methanol constant volume, gets 0.5ml subsequent filtrate in 10ml volumetric flask after filtration, and methanol constant volume is to scale, and at 284nm wavelength place, trap is 3.058, and what be greater than pharmacopeia regulation is not less than 0.40.

Method of quality control research

(1) character

Compound dragon's blood gel (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1) is the red viscous body of homogeneous exquisiteness.

(2) pH value

Get compound dragon's blood gel 1g, add the distilled water 10ml newly boiling, after ultrasonic mixing, recording pH value is 6.88.

(3) check

Centrifugal test is got 3 crowdes of each 10g of compound dragon's blood gel and is put in centrifuge tube, after the centrifugal 30min of 3000r/min, takes out, and gel is without lamination.

Heat resistant test is got 3 crowdes of each 20g of compound dragon's blood gel and is put in 50ml beaker, 55 DEG C of water-bath 2h, and gel is without lamination and change color.

Low temperature resistant test is got 3 crowdes of each 20g of compound dragon's blood gel and is put in 50ml beaker, places 72h for-18 DEG C, and gel is without lamination and change color.

Determining of content assaying method

Chromatography condition

The selection of mobile phase: investigate respectively 1% acetic acid-acetonitrile different proportion (63:37,64:36, mobile phase 69:31), result is taking 1% acetic acid-acetonitrile (64:36) as mobile phase, 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one in test sample, lourerin B peak separating effect the best and disengaging time are suitable, therefore select 1% acetic acid-acetonitrile (64:36) for mobile phase.

Detect the selection of wavelength: the result of investigating according to Literature Consult and test, determine that chromatographic condition is: mobile phase: 1% acetic acid-acetonitrile (64:36), detects wavelength: 275nm, flow velocity: 1.0ml/min.30 DEG C of column temperatures.Theoretical cam curve is pressed lourerin B and is calculated, and is not less than 5000.

The preparation of reference substance solution: precision takes 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one 2.0mg, lourerin B 1.9mg is as in 50ml volumetric flask, and dissolve with methanol obtains 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one that concentration is 0.04mg/ml and the lourerin B standard substance mixed solution of 0.038mg/ml.

Gel extraction method is determined

Extract the selection of solvent: take 3 parts of compound dragon's blood gels, every part of 2g, add respectively 8ml ethyl acetate, n-butyl alcohol, methanol, is settled to 10ml after ultrasonic 30min, and HPLC measures 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one lourerin B, result shows that separating effect, peak shape and the content of methanol are best, so adopt methanol to carry out supersound extraction.

The selection of extraction time: get 3 parts of 2g compound dragon's blood gels, add 8ml methanol ultrasonic 20min, 30min, 40min respectively, prepare need testing solution, assay result shows: there is impact the supersound extraction time on compound dragon's blood gel content, ultrasonic 30min and 40min content no significant difference, but far above 20min's.For ensureing to extract completely and shortening the processing time, adopt supersound extraction 30min.

The preparation of need testing solution

Get compound dragon's blood gel 2g in 10ml volumetric flask, add 8ml methanol, ultrasonic 30min, lets cool rear methanol constant volume, shakes up, and filters, and gets subsequent filtrate, to obtain final product.

Specificity test

Get the raw material except Sanguis Draxonis according to prescription, make negative control sample by compound dragon's blood preparation technology, preparation method according to above-mentioned need testing solution is prepared negative fluid, measure according to said method, result show 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B appearance time with negative control product appearance time without overlapping, show that substrate do not disturb 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B assay, the method specificity is good, the results are shown in Figure 1,2,3.

Methodological study

Precision test: precision takes 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, lourerin B reference substance is appropriate, adds methanol and makes to dissolve, and according to above-mentioned chromatographic condition, sample size is 20ul, and continuous sample introduction 6 times, records peak area.The RSD of result 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, lourerin B is respectively 0.687% and 0.904%, shows that this method precision is good, the results are shown in Table 34.

Table 34 Precision test result

Reference substance is linear to be investigated

Precision takes 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one 3.0mg, lourerin B 3.1mg in 25ml volumetric flask, is settled to 25ml and obtains the lourerin B that 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one that concentration is 0.12mg/ml and concentration are 0.124mg/ml after methanol ultrasonic dissolution.Draw respectively 4ml, 2ml, 1.5ml, 1ml, 0.5ml, 0.2ml, in 10ml volumetric flask, adds methanol and is diluted to scale, shakes up.Sample size is 20ul, and injection liquid chromatography is measured according to above-mentioned chromatographic condition, records chromatographic peak area.SPSS17.0 software data processing, taking peak area (Y) as vertical coordinate, reference substance sample size (X) is abscissa, drawing standard curve, calculating regression equation is: ya=136.229xa-0.21(r=0.9999) yb=74.574xb-0.88(r=0.9999), result shows, 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one is good in 0.0004mg/ml～0.04mg/ml scope internal linear, and lourerin B is good in 0.00038mg/ml～0.038mg/ml scope internal linear.

Replica test

Get 6 parts of same batch of compound dragon's blood gels, every part of 2g, prepares test sample according to the method described above, measures the peak area of 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B, and calculates RSD value and be respectively 0.91%, 1.07%, shows that the repeatability of the method is good, the results are shown in Table 35.

Table 35 replica test result

Sample stability test

Get same batch of compound dragon's blood gel 2g, prepare according to the method described above test sample, test sample is placed under room temperature, respectively at the 0th, 1,2,4,6,8,12 hour, draw 20ul and inject high performance liquid chromatograph, record peak area.Result shows that test sample is stable in 12 hours, and the peak area RSD value of 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B is respectively 0.91%, 0.86%, the results are shown in Table 36.

Table 36 stability test result

Application of sample recovery test

Get compound dragon's blood gel 2g and in 10ml volumetric flask, add 8ml methanol ultrasonic dissolution, after letting cool, be settled to scale.After filter paper filtering, get 3 1ml subsequent filtrates in test tube, labelling application of sample is high, medium and low.Add respectively with sample in 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, lourerin B content 120%, 100%, 80% standard substance 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, after lourerin B 1ml fully mixes, draw 20ul inject high performance liquid chromatograph.Record peak area, measure its response rate and be all greater than 95%, and RSD% value is all less than 1%, show that the method average recovery is good, the results are shown in Table 37.

Table 37 application of sample recovery test result

Sample size is measured:

According to above-mentioned content assaying method, measure the content of 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B in 2 batches of compound dragon's blood gels and be respectively 0.004210mg/ml, 0.007201mg/ml.

Experiment shows: compound dragon's blood gel quality affects control method is: and Thermo BDS HYPERSIL C18 chromatographic column (250*4.6nm, 5 μ are m); Mobile phase: 1% acetic acid-acetonitrile (64:36), detects wavelength: 275nm; Flow velocity: 1.0ml/min; 30 DEG C of column temperatures.The separating degree of Sanguis Draxonis gel 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B chromatographic peak is all greater than 1.5 with this understanding, and tailing factor is between 0.95-1.05, and theoretical cam curve is all greater than 5000, meets Pharmacopoeia of the People's Republic of China requirement.3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one is good linear relation within the scope of 0.0004mg/ml～0.04mg/ml, and regression equation is ya=136.229xa-0.21, r=0.9999; Lourerin B is good linear relation within the scope of 0.00038mg/ml～0.038mg/ml, and regression equation is yb=74.574xb-0.88, r=0.9999; Its precision RSD is respectively 0.687%, 0.904%; Stability RSD is respectively 0.91%, 0.86%; Repeatability RSD is respectively 0.91%, 1.07%; Average recovery rate is respectively 99.0%, 99.8%, and RSD is respectively 0.10%, 0.80%.

Embodiment 8

The investigation of explanation vitro release as an example of compound dragon's blood (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1) example

Release medium: three layers of gauze

Disperser and medium: adopt TK-12B type transdermal instrument, diffusion cell area 2.92cm2, acceptance pool volume is 8.0mL, diffusion cell temperature is 37 ± 0.2 DEG C, the rotating speed of magnetic stick is 200 ± 5r ﹒ min-1, and dispersive medium is 95% ethanol that shifts to an earlier date 37 DEG C of constant temperature water baths.

Dissolution test method: get three layers, gauze, be fixed on diffusion cell, tighten, directly take 2g1.2% compound dragon's blood gel on gauze, gel is evenly smeared on gauze with waterproof paper flicking, acceptance pool is filled it up with dispersive medium, with 37 ± 0.2 DEG C of water bath heat preservation magnetic sticks, speed with 200 ± 5r ﹒ min-1 stirs, respectively at 0, 3, 6, 10, 15, 20, 50, 90, 120, 180min sampling, take out whole liquid 8mL in acceptance pool, add immediately the dispersive medium of 37 DEG C of preheatings of equivalent, add fashionable attention and drain bubble, acceptable solution is collected in the test tube of having numbered for subsequent use.

Because the content of 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B is little in Sanguis Draxonis, should not directly survey 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and the lourerin B content in acceptable solution with HPLC, adopt concentrated acceptable solution, after 2ml dissolve with methanol, measure.

The processing of result is tried to achieve accumulative total infiltration capacity according to following formula

Q_{n} = \frac{Σ_{i = 1}^{n} C_{i} \times 8}{2.92}

Wherein Ci is i the drug level that sample point records.With accumulation infiltration capacity, Q maps to time T, after drug osmotic reaches stable state, and the scalp speed that wherein slope of straight line portion is medicine, i.e. steady-state permeation speed (Jss).The infiltration coefficient (Kp) of medicine calculates with following formula, the drug level that C is supply pool

Kp=Jss/C

Statistical method

Experimental data adopts SPSS17.0 software to carry out statistical analysis, mean ± standard deviation for measurement data (X ± SD) represents, difference between more each group of alone factor variance analysis (One-way ANOVA), P < 0.05 has statistical significance for difference.

Vitro release is investigated and be the results are shown in Table 38.

The average release result (n=4) of table 38 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B

By table 38, Fig. 6 is known, and in compound dragon's blood gel, 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one average release in 180min reaches 96.13%, and from 50min, release profiles tends to balance.After 90min, release changes not quite, kept stable, and lourerin B average release in 180min reaches 92.94%, 30min left and right release profiles and tends to balance, and after 50min, release changes not quite, kept stable.

Claims

1. Sanguis Draxonis/Sanguis Draxonis compound recipe, it consists of the Sanguis Draxonis of weight portion: Borneolum Syntheticum 9:1 or Sanguis Draxonis: Borneolum Syntheticum 9:1.

2. a Sanguis Draxonis compound recipe, it consists of the Sanguis Draxonis of weight portion: berberine: Borneolum Syntheticum 9:4:1 or Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.

3. a Sanguis Draxonis compound recipe, it consists of the Sanguis Draxonis of weight portion: berberine: Borneolum Syntheticum 9:4:1 or Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.

4. use the compound recipe of claim 1 or 2 or 3 as gel, liniment, cataplasma, bandage, preparation capable of permeating skin, patch.

5. the application of the compound recipe of claim 1 or 2 or 3 in preparation treatment more wound, tissue repair medicine.

6. application as claimed in claim 5, is characterized in that described medicine is for treatment wound, blood stasis wound, suppurative wound, dermatitis, comedo, vasculitis, cirsoid medicine.

7. a gel, it is characterized in that getting the compound recipe of claim 1 or 2 or 3, its compound medicine weight is 0.15g, with mixing of 2.5mL propylene glycol, add carbomer 9.85g, with triethanolamine adjusting, pH value is 7.0, and gained gel has glossiness, stretchability, uniformity and the centrifugal property of preparation requirement.

8. a liniment, is characterized in that getting the compound recipe of claim 1 or 2 or 3, takes this compound medicine 0.2g, is repeatedly dissolved in propylene glycol 1.0g on a small quantity, makes it fully dissolve the phase as A; Get 4% polyvinyl alcohol 8.0g, add sodium carboxymethyl cellulose 0.2g and polyvinylpyrrolidone 0.5g, 70 DEG C-80 DEG C of water-baths, make the abundant swelling dissolving of adjuvant as B phase; B is added to the A mixing that stirs in mutually, finally add surfactant tween 80 to keep the stability of liniment, liniment is evenly spread upon on the glass plate that scribbles liquid paraffin, 37 DEG C of oven dry in baking oven, the outward appearance of gained liniment has uniformity, stretchability, mobility and film pliability.

9. a cataplasma, is characterized in that getting propylene glycol-water mixed solvent of appropriate carbomer 44:56, obtains 20g3% carbomer after swelling, adds therein 50g2% aqueous tartaric acid solution, fully mixes, for subsequent use; The compound medicine 6g that gets claim 1 or 2 or 3, uses 15g propylene glycol: the mixed solvent of 95% ethanol 3:12,50 DEG C of water-baths are dissolved, for subsequent use; Get sodium polyacrylate 4g, CMC-Na2g, aluminium hydroxide 0.3g, EDTA0.5g add in beaker, add 22g glycerol to mix well, for subsequent use; Medicine after dissolving is added to the mixed phase such as sodium polyacrylate-glycerol, after fully mixing, carbomer-tartaric acid is added rapidly wherein, stir rapidly, the local caking of control, then continue to stir 60 minutes, treat mastic deliquescing, granular sensation is coated with while disappearance, and room temperature is placed after 3 days and be get final product.