CN103919967B - Sanguis Draxonis compound recipe and the application in preparation treatment more wound medicine thereof - Google Patents

Abstract

The invention provides and a kind of become the compound preparation being grouped into other by having Sanguis Draxonis or Sanguis Draxonis that treatment more wound, tissue repair, vasculitis etc. act on, consisting 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.Their preparation method and the application in preparing the preparations such as compound gel, liniment, cataplasma, bandage, preparation capable of permeating skin (such as patch) of the Sanguis Draxonis compound recipe thereof.Sanguis Draxonis, Sanguis Draxonis compound preparation and two kinds of positive drug (Madecassol, JINGWANHONG) are compared, and are respectively provided with wound healing, the tissue repair effect of being obviously promoted, and anxious poison and long term toxicity show that said preparation uses safety.

Description

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

Technical field:

The invention belongs to technical field of pharmaceuticals, in particular it relates to Sanguis Draxonis or medicaments compound system that Sanguis Draxonis is main component Agent, prepares 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 warp, have blood circulation promoting and blood stasis dispelling, astringing to arrest bleeding, The remarkable efficacy such as hard masses softening and resolving, promoting tissue regeneration and ulcer healing, is usually used in inside and outside all sections, such as: traumatic injury, blood stasis, hemorrhage, carbuncle ulcer, skin ulcer The diseases such as the long disunion in face, are many ancient prescriptions and the important composition medical material of Empirical formula, are called " panacea invigorated blood circulation " prompting Sanguis Draxonis There is repair in trauma effect.The medicinal Sanguis Draxonis that China uses the earliest " comes from the Western Regions ", and its Original plant is to originate in West Asia and north African Agavaceae plant Folium Dracaenae cambodianae (Dracaena draco).Bright clear since, Indonesia and Malaysia Palmae Caulis Fibraureae Platymiscium Sanguis Draxonis (Daemonorops dracoBL.) produced red resin becomes the main source of the medicinal Sanguis Draxonis of China, i.e. " Sanguis Draxonis " that the Pharmacopoeia of the People's Republic of China 2010 editions records (the resin from Daemonorops draco, RDD, Referred to herein as Sanguis Draxonis).Last century the seventies, famous botanist Cai Xitao professor be found to secret out of redness in Yunnan Province The Agavaceae dracaena plant Dracaena cochinchinensis (Dracaena cochinchinensis Lour.S.C.Chen) of resin, Successfully develop domestic Dragon Blood, listed in 1999 " national drug standards " promulgated by the ministries or commissions of the Central Government (WS3-082 (Z-016)-99 (Z)), name For Sanguis Draxonis (the resin from Dracaena cochinchinensis, RDC), thus it it is the state of famous and precious Chinese medicine Resina draconis Productization opens approach.But, domestic Sanguis Draxonis (RDC) and import Sanguis Draxonis (RDD, i.e. Sanguis Draxonis) plant section belong to difference, chemistry There were significant differences for composition.Sanguis Draxonis and Sanguis Draxonis (that is: Sanguis Draxonis) are all used clinically for blood circulation promoting and blood stasis dispelling, promoting tissue regeneration and ulcer healing at present, but Two kinds of Sanguis Draxonis lack basic research in terms of more wound field medicinal effects, and whether Sanguis Draxonis can substitute Sanguis Draxonis for the more side of wound Face also has no report.For the chemical constitution study of Sanguis Draxonis, the result obtained from different Original plants is not quite similar.Sanguis Draxonis chemistry The complexity of composition makes the determination of its active component particularly difficult.It is confirmed to be with monomeric form at present and more creates active component Only from Euphorbiaceae Fructus Crotonis platymiscium Croton salutaris, the alkaloid being separated in the produced Sanguis Draxonis of C.lechleri taspine.The chemical research of Sanguis Draxonis is thought that its main component is flavonoid and phenols by domestic scholars, and import Sanguis Draxonis is main Containing polyphenolics and ter penoids, but both are all without taspine.The modern pharmacology research of Sanguis Draxonis has been focused into it Using blood system.Zoopery shows, Sanguis Draxonis can reduce packed cell volume, whole blood viscosity and plasma viscosity, can press down Platelet aggregation processed, expands blood vessel, improves microcirculatory blood flow, embodies the effect of invigorating blood circulation；Clotting time, blood plasma can be shortened multiple simultaneously Calcium time and euglobulin lysis time, and there is heparin resistance, can be used for stopping blooding.The two-ways regulation of invigorating blood circulation-stop blooding of Sanguis Draxonis Mechanism explains it has the treatment of related disorders to make with blood coagulation-haemolysis balance is destroyed coronary heart disease, cerebral ischemia, metrorrhagia etc. With.Tao Yi etc. compares internal, the In Vitro Anti platelet aggregation of Sanguis Draxonis and Sanguis Draxonis, result show both aobvious Writing anticoagulation, Sanguis Draxonis effect is the most excellent.Another of Sanguis Draxonis notable drug effect expelling pus and promoting granulation is also in substantial amounts of clinic Practice is confirmed.For a long time, Sanguis Draxonis is widely used in soft tissue contusion, decubital ulcer, pressure ulcer, sugar at surgery and department of dermatologry Urine foot disease ulcer, varicose ulcer of the lower limb and the treatment of oral ulcer, achieve good effect.This confirms clinically Sanguis Draxonis, Sanguis Draxonis stop blooding-invigorate blood circulation in addition to dual regulation except having, and also have the effect promoting wound healing.

The at present both at home and abroad basic research to wound healing shows, wound healing is divided into three phases: inflammation, organize the formation of And tissue remodeling.Three phases crossover is carried out, and each stage all has specific cytokine and chemokines to participate in, by right The regulating and controlling effect of each stage factor, medicine can with accelerated wound healing process and reduce scar tissue generate.

Sanguis Draxonis is more universal in clinical practice, common based on peroral dosage form on market.Its main dosage form is Capsule and dispersible tablet etc., main pharmacological is blood circulation promoting and blood stasis dispelling.But oral formulations has drug effect plays the time relatively slowly to stomach Intestinal has a zest, liver first-pass effect etc., present clinical treatment wound or decubital ulcer etc. often use Sanguis Draxonis dry powder and Other drug matches or uses the method for ointment of Sanguis Draxonis.So far, prior art there are no with Sanguis Draxonis, blood Exhaust for main formula composition Chinese medicine compound for preparing the report of the application in the medicine treating more wound or tissue repair.

Summary of the invention

In place of overcoming deficiencies of the prior art, present invention aims to patient and comply with taking medicine Property is poor, enables medicine trauma patient be played soon to drug effect, Sanguis Draxonis, Sanguis Draxonis are made gel, liniment, cataplasma, The external preparation such as wound patch, preparation capable of permeating skin (such as patch), are not only reduced the toxic and side effects of medicine but also can be played by local complete Body effect.

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

A kind of Sanguis Draxonis/Sanguis Draxonis compound recipe, consisting 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, consisting of the Sanguis Draxonis of weight portion: berberine: Borneolum Syntheticum 9:4:1 or Sanguis Draxonis: Rhizoma Coptidis Element: Borneolum Syntheticum 1:1:0.15.

A kind of Sanguis Draxonis compound recipe, consisting 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.

Present invention also offers the application in preparation treatment more wound, tissue repair medicine of the above-mentioned compound recipe.

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

Invention also provides a kind of gel, take the above-mentioned compound recipe of the present invention, its compound medicine weight is 0.15g, With the mixing of 2.5mL propylene glycol, adding carbomer 9.85g, regulate with triethanolamine, pH value is 7.0, and gained gel has system Glossiness that agent requires, stretchability, uniformity and centrifugum.

With, a kind of liniment, take the above-mentioned compound recipe of the present invention, weigh this compound medicine 0.2g, be the most repeatedly dissolved in the third two In alcohol 1.0g so that it is fully dissolve as A phase；Take 4% polyvinyl alcohol 8.0g, add sodium carboxymethyl cellulose 0.2g and polyethylene Ketopyrrolidine 0.5g, water-bath 70 DEG C-80 DEG C, make the abundant swelling dissolving of adjuvant as B phase；It is added to B in A phase stir Mixing, is eventually adding surface active agent tween-80 and keeps the stability of liniment, liniment uniform application is being scribbled liquid On the glass plate of paraffin, 37 DEG C of drying in baking oven, the outward appearance of gained liniment has uniformity, stretchability, mobility and film Pliability.

And, a kind of cataplasma, take the propylene glycol-water mixed solvent of appropriate carbomer 44:56, swelling after obtain 20g3% carbomer, adds 50g2% aqueous tartaric acid solution wherein, fully mixes, standby；Take the above-mentioned compound medicine of the present invention 6g, uses 15g propylene glycol: the mixed solvent of 95% ethanol 3:12,50 DEG C of water-baths are dissolved, standby；Take sodium polyacrylate 4g, CMC- Na2g, aluminium hydroxide 0.3g, EDTA0.5g add in beaker, add 22g glycerol and mix well, standby；After dissolving, medicine adds poly- After the mixed phases such as sodium acrylate-glycerol, fully mixing, carbomer-tartaric acid is rapidly added wherein, stirs rapidly, anti- Controlling local caking, be further continued for stirring 60 minutes, treat mastic deliquescing, granular sensation is coated when disappearing, and room temperature after placing 3 days is ?.

The present invention relates to Sanguis Draxonis (or Sanguis Draxonis) compound preparation, consisting of Sanguis Draxonis (or Sanguis Draxonis): Borneolum Syntheticum 9:1, sanguis draconis Exhaust (or Sanguis Draxonis): berberine: Borneolum Syntheticum 9:4:1, Sanguis Draxonis (or Sanguis Draxonis): berberine: Borneolum Syntheticum 1:1:0.15.This compound recipe is healed in preparation Application in wound or tissue repair medicine, in treatment wound, blood stasis wound, suppurative wound, dermatitis, comedo, vasculitis Deng the application in 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 (such as patch).Present invention research shows: this sanguis draconis Exhaust (Sanguis Draxonis) compound preparation and can promote mice, rat, the healing of rabbit back skin injury wound.Decrustation time, healing time divide Substantially not shortening, display Sanguis Draxonis has promotion wound healing, accelerates wound incrustation, decrustation effect；Experiment it was additionally observed that dragon Sanguis Draxonis group scar tissue is less, and site of injury hair growth recovers good.

Accompanying drawing illustrates:

Fig. 1 is negative controls chromatogram；

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 on rat wound surface surface and wound surface is red and swollen upon administration, infect, edge of wound new life epidermal hyperplasia feelings Condition.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, 8E For bare substrate group, 8F is Mei Jiasi group, and 8G is JINGWANHONG group, and 8H is pure blank group.

Detailed description of the invention:

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

Embodiment 1

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

Compound dragon's blood shows in mice wound test model and necessarily promotes incrustation, the therapeutical effect of decrustation healing, The pharmacological action with Sanguis Draxonis itself with putrefaction-removing granulation-promoting wound healing matches.Rhizoma Coptidis has pathogenic fire purging solution in this prescription Poison, effect of clearing heat and moistening dryness, can make traumatic infection rate reduce, and it is compound dragon's blood prescribed treatment that the wound initial stage local of minimizing is oozed out One of effect.Because the protective layer due to skin surface is destroyed in open wound, antibacterial easily invades so that wound Easily cause infection, so wound is had astriction in the case of wound surface holding is the most infected to reduce wound face simultaneously Long-pending.Borneolum Syntheticum has refrigerant, minimizing pain effect.Select three kinds of medicines composition compound dragon's blood medicine 1 group (Sanguis Draxonis or Sanguis Draxonis: yellow Lian Su: Borneolum Syntheticum 1:1:0.15), compound dragon's blood medicine 2 groups (Sanguis Draxonis or Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1), determine the two Effect to tissue repair.

Selecting 40 Kunming mouses, the laboratory animal department of the Chinese Academy of Sciences of Kunming Medical University provide, body weight is 22～25g, freely Drinking water and ingest, being randomly divided into 4 groups, often 10 white mice of group, lumbar injection pentobarbital sodium 40mg/Kg anaesthetizes, back unhairing, 70% ethanol routine disinfection, spreads aseptic towel, cuts the skin of a diameter of 1.2cm size at back, deep and fascia muscle, and 4 groups respectively Give different disposal: positive controls, blank group, compound dragon's blood 1 group and compound dragon's blood 2 groups.Positive controls is coated with crin Mycin lidocaine gel agent, blank group is outer is coated with gel substrate, the other externally coated compound dragon's blood medicine of drug component 1 group, compound recipe Dragon's blood medicine 2 groups.All wound surface are smeared by equal direct drug injection cotton, and thickness is about 1mm, all should be completely covered by medicine every time Wound surface, often group is 1 cage.Wound surface every day, coating 2 times, respectively at postoperative 1 day, 3 days, 6 days, 9 days, 12 days white mice to often organizing After the method recording medicine of the digital employing digital photographing carrying out wound area, the wound area of 3,6,9,12 days, uses image soft Part analyzes wound surface area.Wound area before medication is deducted the wound area after medication than the wound area before medication, obtain Ratio be wound surface skin healing percentage rate, and calculate beginning decrustation natural law and healing days, tested as an evaluation of result The effect of medicine.After last medication, next day monitors rat serum picture, then takes off neck and puts to death, and clip wound site skin is fixed on 10% In formalin, specimens paraffin embedding slices, HE dyes, and basis of microscopic observation respectively organize the epidermis reparation, very of burned rats skin Materials with hide glue fibril Shu Xiufu, dermic hair follice hypertrophy, corium and the healing of subcutaneous tissue inflammatory cell infiltration.

The postoperative systemic conditions of all animals is good, without untoward reaction such as diarrhoea, refusing to eats, after second day after operation anesthetics releases Spirit is normal, diet normal, and after modeling, all wound surface all have hemorrhage, tissue fluid to have to ooze out and peripheral tissue edema.Postoperative immediately Coating, at the 2nd day wound, no blood flows out, and skin trauma mouth is dry hardening, and spirit and diet are normal.At about the 5th day Wound mouth begins with thicker crust and covers in wound, substantially alleviates without wound surface peripheral tissue edema, and secretions is less.12nd After it, wound surface starts decrustation, and wound is neoplastic skin, and hair increases, but still has uncovered skin.

Wherein positive control drug, blank group, medicine 1 group, medicine 2 groups are respectively to scab forming time, decrustation time and healing rate Compare between healing time group, the results are shown in Table 1 and table 2.

Wound area slip tested by table 1

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

Table 2 is formed a scab, decrustation and healing time

Group	Scab forming time (d)	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

Histopathological scores result

Wounds in mice pathology is cut by wounds in mice pathological section histopathological scores establishment of standard and different pharmaceutical group Sheet is marked, respectively such as table 3 below and table 4.

Table 3 histopathological scores standard

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

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 compares with blank, and compound dragon's blood gel of the present invention, positive control drug are in wound Hinder healing rate aspect and have preferable pharmacological effect.Wherein compound dragon's blood medicine 2 groups (Sanguis Draxonis or Sanguis Draxonis: berberine: Borneolum Syntheticum 9: 4:1) effect becomes apparent from.After test finding, trauma model is built up, mouse skin and a small amount of tissue adhesions of Musclar layer.After 2 days The skin of wound site is hardening, and no blood flows out, and diet is normal with mental status.After 5 days, wound site begins with reddish black blood Crust covers, and after decrustation, wound area substantially diminishes, and wound location is cerise, around a small amount of hair growth of healing skin.Wound After healing completely, hair increases also with the reduction of wound area.After wound heals completely, hair continued growth continues in wound Continuous wound coverage position.

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；

Positive control drug Mei Jiasi (Madecassol) anti-scar ointment: 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, male and female half and half, 6～8 week old, body weight 180～220g, it 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.Rearing conditions: temperature 25～27 DEG C, humidity 70 ± 10 DEG C, animal freely ingests drink Water.

After laboratory animal is weighed, anaesthetize according to body weight (30mg/Kg), after anaesthetizing, be fixed in plank, with shaving Hair device carries out shaving 3cm × 3cm, carries out wound and prepares.

The modeling test tube of a diameter of 2cm dips povidone iodine, is buckled in rat back (left and right sides outside spinal column about 2cm), uses Toothed forceps mentions superficial skin, 45 ° and contact skin, shears, deep and subcutaneous.Successful for modeling rat is randomly divided into 6 Group, often group 7, individually raises.

Being administered first group is Sanguis Draxonis low dose group, and second group is Sanguis Draxonis high dose group, and the 3rd group is Sanguis Draxonis low dosage Group, the 4th group is Sanguis Draxonis high dose group, and the 5th group is bare substrate group, and the 6th group is Mei Jiasi group, and the 7th group is JINGWANHONG group, 8th group is pure blank group.Respectively organizing by said medicine and be administered every day, gauze covers, and it is fixing that medical adhesive tape carries out wrapping.The most every Wound is retouched meter by it, takes pictures.

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

The preparation tissue of tissue slice is fixed through 4% paraformaldehyde, carries out gradient wine successively after the PBS eluting of 0.1M three times Loss of essence water, dimethylbenzene is impregnated with and paraffin embedding, section, carries out HE routine histologic dyeing

Observation index and the method for analysis

Situation and upon administration is oozed out on perusal rat wound surface surface, and wound surface is red and swollen, infect, edge of wound new life epidermis increases Raw situation, and record wound healing time.After wound, 3d starts to calculate Wound healing rate, uses transparent template tracing to measure Local woanded surface area, is calculated by following equation: healing rate=(original wound surface area-existing wound surface area)/original wound surface area × 100%, with healing rate >=95% for healing completely, observe and record each group of healing time.

Experimental result

1. wound healing rate

After wound, 1d naked eyes are barely perceivable wound surface change, therefore go out the data of 1d, dragon in same time shop Sanguis Draxonis group, Sanguis Draxonis group and Madecassol group, the Wound healing rate of JINGWANHONG group do not have obvious difference, but are above blank group With bare substrate group, difference statistically significant (P ＜ 0.05), the results are shown in Table 5.

Table 5 trauma in rat different time healing rate compares (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 has a notable difference, but all Than blank group and bare substrate group time period, difference statistically significant (P ＜ 0.05), the results are shown in Table 6.

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

Using SPSS17.0 to carry out statistical disposition, low dosage Sanguis Draxonis, low dosage Sanguis Draxonis and two kinds of positive drug compare, P ＞ 0.05, it is not significantly different from, illustrates that low dosage Sanguis Draxonis and Sanguis Draxonis all have the effect promoting wound healing, Sanguis Draxonis can simultaneously To substitute Sanguis Draxonis；Low dosage Sanguis Draxonis, Sanguis Draxonis and two kinds of positive drug compare with bare substrate group, blank group, P ＜ 0.05, tool There were significant differences；The Sanguis Draxonis of low dosage, Sanguis Draxonis compare with Sanguis Draxonis, the Sanguis Draxonis of high dose, and P ＜ 0.05 has significant difference, And low dosage effect is better than high dose；High dose Sanguis Draxonis, Sanguis Draxonis compare with bare substrate group, blank group, and P ＞ 0.05 does not has Significant difference.

Rabbit wound healing assay

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

Positive control drug: Mei Jiasi (Madecassol) anti-scar ointment: 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 do not limit, body weight 3～3.5Kg, are purchased from Kunming Medical University's laboratory animal The department of the Chinese Academy of Sciences, the quality certification number: SCXK (Yunnan) 2011-0004.Rearing conditions: temperature 25～27 DEG C, humidity 70 ± 10 DEG C, animal is freely taken the photograph Food drinking-water.

It is grouped after 12 rabbit weigh, is randomly divided into 2 groups according to body weight, often group 6.

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

It is administered first group of 6 rabbit, gives low dosage Sanguis Draxonis, low dosage Sanguis Draxonis, bare substrate and Madecassol respectively Positive drug；Second group of 6 rabbit, gives high dose Sanguis Draxonis, high dose Sanguis Draxonis, JINGWANHONG positive drug and pure blank respectively.By upper Stating medicine and respectively organize administration every day, gauze covers, and it is fixing that medical adhesive tape carries out wrapping.Every day wound is retouched meter simultaneously, take pictures.

Observation index and the method for analysis

Situation and upon administration is oozed out on perusal rabbit wound surface surface, and wound surface is red and swollen, infect, edge of wound new life epidermis increases Raw situation, and record wound healing time.After wound, 3d starts to calculate Wound healing rate, uses transparent template tracing to measure Local woanded surface area, is calculated by following equation: healing rate=(original wound surface area-existing wound surface area)/original wound surface area × 100%, with healing rate >=95% for healing completely, observe and record each group of healing time.

More than experiment is reached a conclusion, compound dragon's blood (Sanguis Draxonis) gel and positive control drug and blank wound healing Rate aspect has preferable pharmacology effect compound recipe to answer.Wherein healing rate size is respectively compound dragon's blood (Sanguis Draxonis): Borneolum Syntheticum gel, multiple Side's Sanguis Draxonis: Rhizoma Coptidis: Borneolum Syntheticum gel, lincomycin hydrochloride and lidocaine hydrochloride gel agent, blank group.Scab forming time, de- It also it is above result on crust time and healing time.After in experiments it is found that trauma model is built up, mouse skin and Musclar layer A small amount of tissue adhesions.After 2 days, the skin of wound site is hardening, and no blood flows out, and diet is normal with mental status.Wound after 5 days Position begins with reddish black blood crusts and covers, and after decrustation, wound area substantially diminishes, and wound location is cerise, around healing skin A small amount of hair growth.Wound heal completely after hair also with wound area reduction increase.After wound heals completely, hair continues Wound coverage position is continued in the continuous wound that is grown in.

Embodiment 2

Prepared by compound dragon's blood (Sanguis Draxonis) gel

This experiment use by carbomer P940 in water the most swelling after, add drug to solution and be sufficiently mixed, three second Hydramine regulation pH value makes gel reach bigger stickiness denseness.Gel should have preferable glossiness, stretchability, uniformity With centrifugum.

1. solvent species is investigated

Determining the ratio of three kinds of medicines in compound dragon's blood prescription according to document and clinical experience, compound recipe is with dragon's blood medicine It is main.Sanguis Draxonis has water insoluble, ether and dilute acidic solution, is easy to become solution in the physicochemical property of methanol, ethanol and basic solvent. The com-parison and analysis different solvents dissolving situation to compound medicine, Selective dissolution is good and solvent non-irritating to human body is as multiple The lytic agent of side's dragon's blood medicine.

2. drug solubility is investigated by solvent

Precision weighs 5 parts of compound dragon's blood medicated powder, every part of 100mg, is dissolved in 5mL glycerol, propylene glycol, Polyethylene Glycol respectively 200,75% ethanol, 95% ethanol, is sufficiently stirred for mixing, observes medicine dissolution situation, the results are shown in Table 7 after placing 5 minutes.

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

Remarks: " ++ " is completely dissolved, "+" it is slightly soluble, "-" is the most insoluble

As can be seen from the above results, compound dragon's blood medicine can be completely dissolved in propylene glycol and 75% alcohol solvent. The ethanol of 75% has bigger zest to skin, particularly obvious to zest at skin trauma, easily causes wound pain, Open wound patient should not use, it is impossible to uses in a large number as drug solvent.Molten as compound dragon's blood medicine of propylene glycol Solve agent not only little but also there is Penetration enhancing effect to human body skin zest, make medicine easily through skin be absorbed by organisms enough after send out Wave general action.

3. propylene glycol consumption is investigated

Take 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, is thoroughly mixed, and observes after 15min, can be by when solvent volume is 15.0mL Medicine fully dissolves, and drug solution spreads upon nonirritant on skin, preferable without granular sensation, lubricity.

4. gel-type vehicle is investigated

For enabling carrier matrix to mix homogeneously with medicine, through 3 kinds of water-soluable gel substrate carbomers, alginate and bright Glue compares, and occurs without situations such as layering, caking, medicine precipitations using carbomer as substrate in medicine adition process, and And the gel of preparation becomes that colloidality is good, stable in properties and preparation method simple.Select the carbomer gel solution of actual concentrations so that it is Can preferably compatibility absorb the drug.Taking after compound dragon's blood medicine 1.55g propylene glycol fully dissolves, being added separately to concentration is 0.5%, 0.6%, 0.7%8.45g carbomer gel solution is sufficiently stirred for mixing, observes after standing 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, compound dragon's blood medicine mixes all with the carbomer gel substrate that concentration is 0.6% Even, in adition process Chinese medicine and substrate, there is preferable compatibility, separate out and caking phenomenon without layering, medicine.

5. the investigation of drug loading

The maximum drug loading of ordinary gel agent typically about 10%, but for ingredient its drug loading of difference also Can change.Gel gross weight is fixed as 10.0g, investigates the substrate maximum drug loading to compound dragon's blood medicine.Point Not taking compound dragon's blood medicine 0.15g and 0.30g, propylene glycol fully dissolves, with three after the carbomer substrate of addition equivalent weight Ethanolamine regulation gel pH value, to 7.0, is eventually adding tween 80 to regulate stability and the lubricity of gel, is shown in Table 9.

Table 9 drug loading investigation table

Produce the gel of different prescription proportioning according to above prescription substrate, manufacturing process is observed the dissolving of medicine After effect and addition carbomer substrate, whether preparation can be layered, separate out and caking phenomenon, observes gel after regulation pH value 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, solidifying The outward appearance of colloid, uniformity are preferable, smear without granular sensation on skin, add carbomer 9.85g triethanolamine and regulate pH value Being 7.0, gel is uniform, non-block phenomenon occurs.Compound dragon's blood medicine weight is increased to 0.30g, adds carbomer 9.70g, triethanolamine regulation pH to 7.0, gel is layered immediately, and has block in gel, granular sensation it is obvious that Being difficult to uniform application on skin, stickiness is relatively low.Compound dragon's blood medicine content is respectively adjusted to 0.8%, 1.5%, 2.0%, works as load When dose is 1.5%, the glossiness of preparation, the uniformity and stretchability all reach the appearance requirement of gel, gel when 2.0% Agent medicine content is big, and glossiness substantially reduces, and granular sensation is substantially smeared more difficult, the experiment proved that compound dragon's blood gel Maximum drug loading 1.5%.

6.pH value is investigated

Using the volume of carbomer gel matrix weight, drug weight and propylene glycol as constant basis, triethanolamine regulation is solidifying Colloid is different pH value, carries out gel outward appearance and stability in terms of glossiness, stretchability, the uniformity, centrifugal degree four Comprehensive grading, selects appropriate pH, the results are shown in Table 10.

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

Gel is adjusted to different pH value, and result is as follows: occur obvious lamination during pH=6.0, during pH=6.5 The most easy to apply on skin uniformly pH=9.0 is irritant for skin wound, existing without significantly layering during pH=7.0 or 8.0 As, all meet gel prescription from outward appearance, lamination and situation of smearing on skin during pH=8.0.

Embodiment 3

Prepared by compound dragon's blood liniment

This experiment once have selected the Bletilla glucomannan material as liniment, and it had both belonged to the Chinese herbal medicine toxic and side effects for human body Less, and the Pseudobulbus Bletillae (Rhizoma Bletillae) has the effect of astringing to arrest bleeding, detumescence and promoting granulation, wound repairing.It is widely used in treatment hemoptysis clinically, tells The diseases such as blood, traumatic hemorrhage, sore swollen toxin, pulmonary tuberculosis hemoptysis, ulcer haemorrhage.Containing abundant Bletilla glucomannan in Pseudobulbus Bletillae (Rhizoma Bletillae) tuber, in vain Splendid achnatherum glue is the jelly of a kind of faint yellow homogenizing, water soluble, insoluble in ethanol, is that one has multiple bioactive Chinese medicine Material, has been widely used in the production of clinical treatment preparation, has medical applications widely and is worth.The main preparation side of Bletilla glucomannan Method is: (1) decocting bletilla stem block；(2) by pan-fried filtering exudate, centrifugal, concentration；(3) dehydrated alcohol dehydration, decolouring system are added Obtain Bletilla glucomannan.The main component of Bletilla glucomannan is neutral Pseudobulbus Bletillae (Rhizoma Bletillae) heteropolysaccharide, belongs to pyranoid form glucomannoglycan, has stronger stickiness.In vain This special nature of splendid achnatherum both can jointly play pharmacological action with compound dragon's blood medicine, not only has protection to wound mouth and receives Hold back hemostasis, wound repairing, the pharmacological action of detumescence and promoting granulation are also used as the filmogen of liniment, but in manufacturing process Finding that Bletilla glucomannan is complex as the liniment manufacturing process of filmogen, and the yield of Bletilla glucomannan is relatively low, about 3% is left Right.Use Bletilla glucomannan as the filmogen of liniment, after film forming, toughness is poor, denseness relatively low be not easy on skin fixing, If Bletilla glucomannan concentration increased, the most difficult during coating, after being applied to skin, splitting is poor, is difficult to open.When in liniment Moisture evaporation after, in flakey on skin, moistening effect is poor, does not meets the appearance requirement of preparation.So it should be selected Its synthesis macromolecule is as filmogen.

Suitable filmogen and adjuvant is selected by the single factor of liniment is carried out observation analysis.Polyvinyl alcohol As main filmogen, film forming speed not only fast and also after film forming effective toughness strong, quality is soft.If concentration is too Greatly, stretchability is very poor, difficult coating；Concentration little Yi flows, and can not fix at skin.Adjuvant need to be added carry out optimization formulation and improve film Agent technique.

Owing to the drug loading of liniment is less, being suitable for the medicine of low dose, the big solvent of dose is difficult to mix homogeneously, color and luster Degree and the uniformity are the most poor, so selecting correct drug loading is also to need research in liniment manufacturing process.

1. the investigation of filmogen

Being chosen to membrane material and concentration thereof, polyvinyl alcohol concentration is big, and the denseness of liniment becomes big, is difficult to smear and at skin The film formation time on skin surface extends.After the main component of filmogen is determined, add suitable adjuvant and select correct agent Amount, observes outward appearance and the mouldability of liniment.

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

The mouldability of table 11 liniment and drug loading test

In sum, selecting propylene glycol as solvent, the polyvinyl alcohol of 4% is as the main component of filmogen, compound recipe dragon Sanguis Draxonis drug weight is 0.20g.

In order to make liniment have preferable stretchability, uniformity, mobility, need to add other adjuvant to optimize substrate Prescription.

2. liniment substrate formulation optimization and result

The concentration of polyvinyl alcohol is defined as 4%, and compound dragon's blood medicine weight is 0.2g, propylene glycol 1.0g.Add difference Adjuvant 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, gained liniment tough Property and splitting preferable, the uniformity is poor lamination.

(2) concentration is that 5%PVP1g joins as filmogen in polyvinyl alcohol, adds medicine, the one-tenth of gained liniment Film is preferable, and splitting is preferable, but during film, the low viscosity of concentration is poor, is difficult to be coated with on a glass.

(3) starch 2g dissolves and joins as filmogen in polyvinyl alcohol, adds medicine, and gained liniment splitting is relatively Good, but starch is susceptible to gelatinizing phenomenon in the polyvinyl alcohol join dissolving, and the toughness of liniment is poor, stripping process In easily rupturable.

4. polyvinylpyrrolidone 0.5g, sodium carboxymethyl cellulose 0.2g, heating in water bath joins dissolving after fully dissolving Polyvinyl alcohol in, liniment film property, splitting, toughness are preferable.

3. orthogonal test

According to the result of above experiment, filmogen and adjuvant to liniment are further screened, and use orthogonal The method of test, filters out and affects the principal element of film forming and the suitableeest weight.Investigate 4 principal elements affecting mouldability: poly- The weight (D) of the weight (A) of vinyl alcohol, sodium carboxymethyl cellulose (B), polyvinylpyrrolidone (C) and medicine is examined or check, often 3 levels of individual selecting factors, arrange orthogonal test by L9 (34), and factor and level are shown in set out according to above experiment conclusion and set up water Flat factor and orthogonal test table, be shown in Table 12 and table 13 respectively.

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 relatively big, are difficult to be coated with Open and film formation time on skin extends.Sodium carboxymethyl cellulose has the effect of regulation membrane mobility, and dosage increases The denseness then making liniment strengthens, and plasticity strengthens, but toughness reduces.The drug loading of liniment is less, if dose increases, and institute The liniment of system is uneven, and color, the uniformity and mouldability are poor, and outward appearance does not meets liniment requirement.By in liniment After moisture evaporation, obtained liniment peels off on skin that difficulty, fragility is relatively big, poor toughness, nonelastic, be not easy to stretching, nothing Reflecting feel.Add glycerol in manufacturing process and increase pliability and the glossiness of liniment as plasticizer.

4. liniment scoring criterion

Stretchability: liniment is the most spreadable on skin, preparation is fine and smooth, without granular sensation.2.5 are divided into full marks.

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

Pliability: on skin after solvent volatilization, not easy fracture, flexible and elasticity in stripping process, 2.5 be divided into full Point.

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

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

Investigating according to above result of the test, filmogen composition is respectively polyvinyl alcohol, polyvinylpyrrolidone and carboxylic first Base sodium cellulosate, solvent is propylene glycol, and plasticizer is glycerol.According to above experiment investigation result, each composition pair in substrate The stretchability of compound dragon's blood liniment, uniformity, pliability and mobility all have and have a certain impact.Wherein to liniment The dosage that factor is polyvinyl alcohol, polyvinylpyrrolidone and glycerol that mouldability impact is bigger, the change between each factor Relative complex.In order to optimize and filter out the best prescription of liniment further, Orthogonal Experiment and Design is used to optimize system further Standby technique, using the comprehensive grading of presentation quality and mouldability as inspection target, investigates 4 principal elements affecting mouldability: Polyvinyl alcohol weight (A), polyvinylpyrrolidone weight (B), the weight (C) of medicine and amounts of glycerol (D), each selecting factors 2 Individual level, arranges orthogonal test by L8 (24), sets out factor level table according to above experiment conclusion and be shown in Table 14, orthogonal experiments Being shown in Table 15, variance analysis is shown in Table 16.In above prescription, sodium carboxymethyl cellulose is 0.2g, propylene glycol 1.0g, polyvinyl alcohol Concentration be 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, analyzed by directly perceived, due to The weight factor extreme difference of polyvinyl alcohol is relatively big, so the quality of liniment is affected relatively big by the weight of polyvinyl alcohol, and each factor pair The influence degree of liniment outward appearance mouldability is followed successively by A > C > D > B is the weight of polyvinyl alcohol, drug weight, glycerol the most respectively Amount and polyvinylpyrrolidone weight.Best of breed is A1B3C3.

The result of These parameters is carried out variance analysis, table 14 each factor the result that outward appearance mouldability affects difference can To find out, the weight of polyvinyl alcohol is notable (P < 0.05) on the impact of liniment, by F than the dosage that can be seen that polyvinyl alcohol In four factors, the mouldability for liniment affects relatively big, and in manufacturing process, the weight to polyvinyl alcohol needs note especially Meaning, considers the impact of each factor, amounts of glycerol and polyvinylpyrrolidone notable on mouldability impact, therefore selection 4% is dense The polyvinyl alcohol weight of degree is 8.0g, and medication amount is 0.2g, and amounts of glycerol is 0.5g, and polyvinylpyrrolidone is 0.5g.Optimize work Skill route is A1B1C2D1.

6. prescription confirmatory experiment

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

Propylene glycol be 1.0g sodium carboxymethyl cellulose be 0.2g

Empirically recipe quantity weighs medicine, the most repeatedly adds, and is dissolved in propylene glycol 1.0g and makes it fully dissolve as A Phase, is weighed by prescription weight in polyvinyl alcohol upon dissolution, adds sodium carboxymethyl cellulose 0.2g and polyvinylpyrrolidone 0.5g, water-bath about 80 DEG C makes the abundant swelling dissolving of adjuvant as B phase, and be added in A phase stir mixing by B, finally Add surface active agent tween-80 and keep the stability of liniment.Liniment uniform application is being scribbled the glass of liquid paraffin On glass plate, 37 DEG C of drying in baking oven, observe the appearance uniformity of liniment, stretchability, mobility and film pliability and combine Close scoring.

7. checking test and scoring

In order to verify the accuracy of the above results, to guarantee the reasonability of liniment preparation technology, by above-mentioned technique bar Part, arranges to repeat experiment, and result of the test is shown in Table 17.

Table 17 orthogonal confirmatory experiment result

Reached a conclusion as follows by the checking test of above liniment: the liniment of orthogonal test gained is ratio experiment in character In the most out liniment mobility slightly worse, denseness is relatively big, is difficult to the most spreadable, smears preparation thickness not on skin when smearing One.The consumption being possibly due to polyvinylpyrrolidone reduces, and 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.Coating process needs to the greatest extent Amount reduces the generation of bubble, at about 37 DEG C, slide is carried out volatilization and dries, and volatilizees under human body temperature, accelerates film forming speed Degree.

Embodiment 4

Prepared by compound dragon's blood cataplasma

Catablasm base material is mainly made up of adhesive agent, wetting agent, cross-linking agent and viscosifier etc., aqueous to cataplasma of substrate Amount, adhesion, bioavailability, comfort and breathability play mastery reaction.

1. adhesive agent selects

This experiment uses hydrophilic compounds sodium polyacrylate and polyvinyl alcohol separately as the skeleton material of cataplasma respectively Material, after result adds cross-linking agent and wetting agent in sodium polyacrylate, cohesiveness is very big, and substrate is the most agglomerating, application difficulties.Individually Using polyvinyl alcohol as framework material, the easy membranization of stromal surface, viscosity is the least, makes whole base owing to substrate cohesiveness is less Matter denseness reduces, and is little to coating.Using the mixture of polyvinyl alcohol and sodium polyacrylate as the framework material of cataplasma, Repeatedly still having preferable adhesion after skin adhesive, and can delay the gelation rate of sodium polyacrylate, crosslinking time prolongs Long, cohesiveness suitably reduces, and makes coating duration relatively extend, and is conducive to coating.

2. wetting agent selects

Wetting agent has selected glycerol and two kinds of solvents of propylene glycol respectively, compares propylene glycol through experiment and makes mastic as wetting agent Cohesiveness significantly reduce, insufficient formability, and poor in gloss appearance sense and pliability relatively glycerol.Glycerol is permissible as wetting agent Account for the 30%-50% of whole mastic weight, compare through test of many times, when amounts of glycerol accounts for the 45% of total mastic weight, not only have relatively Good glossiness and cohesiveness is preferable, mouldability is preferable.

3. cross-linking agent selects

Cross-linking agent selects tartaric acid and aluminium hydroxide, and soda acid acts on carrier matrix jointly, promotes framework material and bonding Being sufficiently mixed and cross-linking of agent, crosslinking time is rapid.

4. tackifier selects

With sodium polyacrylate as main skeleton filmogen, select polyvinylpyrrolidone and carboxymethyl cellulose respectively Two groups of sodium of element, as the auxiliary material of catablasm base material, test result indicate that polyvinylpyrrolidone experimental group oozes cloth serious without appointing What viscous force, the splitting of sodium carboxymethyl cellulose experimental group is preferably and nothing oozes cloth phenomenon.Sodium carboxymethyl cellulose is suitable as this Tackifier in prescription.

5. cataplasma orthogonal test

Catablasm base material single factor test result is observed, using ooze the overall merit of cloth, skin tracing ability and mouldability as Inspection target, investigation affects 7 factors of mouldability and is respectively as follows: sodium polyacrylate (A), polyvinyl alcohol (B), aluminium hydroxide (C) With the ratio (G) of tartaric acid (D), sodium carboxymethyl cellulose (E), drug weight (F), propylene glycol and medicine, each selecting factors 3 Individual level, arranges orthogonal test by L18 (37), designs factor level according to above experiment conclusion and be shown in Table 18, orthogonal experiment plan Meter is shown in Table 19.

Table 18 factor level table

Table 19 orthogonal test table

Carrying out cataplasma technical study making according to above prescription substrate composition, subject matter and conclusion are as follows: 1) ooze cloth More apparent 2) cataplasma is short for standing time, is likely to be due to medicine and fails full cross-linked with substrate, takes off when patch is peeled off easy on skin Fall medicine.3) amount of cross-linking agent is different, and crosslinking time is the most different, and the amount of cross-linking agent is the biggest, and the crosslinking rate of mastic is fast, and mastic is easy Solidifying.4) adding tartaric acid to should be in a small amount of repeatedly addition mastic, be sufficiently mixed uniformly with mastic, cross-linking effect is good.

Overall merit is carried out through cutaneous sense test and presentation quality after above cataplasma is placed a week, bright for having The cataplasma prescription of aobvious problem is eliminated, and wherein significantly problem is: ooze cloth serious, without any viscous force；The initial bonding strength of substrate Poor, easily come off after skin is pasted repeatedly；Initial bonding strength > cohesiveness stripping process mesostroma be prone to depart from non-woven fabrics.Outward Appearance quality: serious owing to oozing cloth, mastic fails to keep moisture, and mastic is without any viscous force, and mastic mouldability is poor；Pasty consistence Low, mastic can be made during coating easily to flow out, embossability is poor.

6. catablasm base material ratio optimization experiment

Observing through many experiments, in document, Zhang Lei etc.s select for cross-linking agent and forge and the time is to armful arriving, crosslinker ratio With crosslinking time, certain impact is respectively provided with for mouldability, skin tracing ability and the coatability of cataplasma.With film residual, Cohesiveness, holding power and splitting carry out overall merit, investigate 4 principal elements affecting mouldability: propylene glycol (A), carboxylic first Base sodium cellulosate (B), aluminium hydroxide are examined or check with tartaric ratio (C), crosslinking time (D), 2 water of each selecting factors Flat, arrange orthogonal test by L8 (24).Factor level is shown in Table 20, and orthogonal test is shown in Table 21.

Table 20 factor level table

Table 21 orthogonal test table

Cataplasma is made, result aluminium hydroxide: during tartaric acid=1:1, the crosslinking than 2:1 is fast according to above prescription substrate composition Degree speed wants fast, and crosslinking time shortens, and mastic easily coagulates.Crosslinking time should determine that in 10～20min, after the time is more than 25min During mastic coating, colloid condenses in bulk, it is more difficult to spreadable.Above cataplasma is carried out overall merit, residual from oozing cloth situation, mastic The property stayed, holding power and splitting carry out integration test, and using weight when measuring holding power is 500g counterweight, and splitting is 100g weight Code.Cataplasma area is 3cm × 3cm, holding power method of testing: be attached to by cataplasma on smooth rustless steel iron plate, suspended from 500g counterweight, record cataplasma comes off the time.Splitting method of testing: cataplasma is attached on smooth rustless steel iron plate, away from Clamp with clip on the non-woven fabrics of mastic 2cm, counterweight is positioned over below clip, mastic face is become with rustless steel iron plate face The angle of 180 °.The time that record cataplasma completely falls off from corrosion resistant plate.

7. comprehensive evaluation result

No. 1: ooze cloth on a small quantity, cohesiveness is preferable, repeatedly takes off patch, remains without substrate, falls medicine on a small quantity.Holding power 46s, during stripping Between 12s, during stripping, a large amount of substrate come off.

No. 2: oozing cloth situation more substantially, cohesiveness is preferable, initial bonding strength is relatively big, without falling medicine situation.Holding power 115s, peels off Time 234s.

No. 3: have and ooze cloth situation, relatively No. 2 are relatively light, and cohesiveness is preferable, initial bonding strength compared with 1,2 weak.Holding power 133s, splitting time 355s。

No. 4: have and ooze cloth, similar to No. 3, cohesiveness is preferable, stickiness compared with 1, No. 2 little, fall medicine situation the most obvious.Holding power 148s, splitting time 144s, during stripping, a large amount of substrate come off.

No. 5: it is similar to No. 1 to ooze cloth situation, cohesiveness is preferable, and stickiness is similar to 1, No. 2.Fall medicine on a small quantity.Holding power 49s, Splitting 128s, during stripping, a large amount of substrate come off.

No. 6: ooze cloth on a small quantity, initial bonding strength is relatively big, and cohesiveness is preferable, falls medicine on a small quantity.Holding power 88s, splitting time 83s.

No. 7: several nothings ooze cloth, cohesiveness is fine, initial bonding strength compared with 1, No. 2 weak, fall medicine on a small quantity.Holding power 172s, splitting time 65s, a small amount of substrate comes off.

No. 8: ooze cloth on a small quantity, cohesiveness is fine, initial bonding strength and No. 5 similar, fall medicine on a small quantity.Holding power 59s, splitting time 110s, a small amount of substrate comes off.

In sum, through repeatedly repetition test and the adjustment of prescription matrix components ratio, cataplasma to ooze cloth phenomenon bright Aobvious improvement, good forming ability, mastic initial bonding strength and holding power increase.

8. optimize cataplasma preparation technology

In order to cataplasma outward appearance and quality standard being met state-promulgated pharmacopoeia regulation, above cataplasma formulation and technology is entered again Row optimizes, and Main way is as follows:

1) adjuvant selected is the fewest more good, the most not only makes cost reduce, and it can be avoided that because of increasing of adjuvant The drug loading making cataplasma reduces.

2) Simplified flowsheet step, makes cataplasma reduce some unnecessary intermediate links in commercial production journey, reduces The waste of manpower and financial resources.

3) presentation quality and instrument test index meet States Pharmacopoeia specifications, have preferable affinity to human body skin, and skin chases after Casual preferably.

9. framework material is preferred

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

10. adjuvant is preferred

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

2) wetting agent still selects glycerol, owing to medicine can not be completely dissolved in glycerol, so with appropriate propylene glycol by medicine Thing adds a certain amount of glycerol after dissolving, and is possible not only to keep cataplasma surface gloss and wettability, and glycerol is to human body Without any side effects have preferable moisture retention, and after repeatedly pasting, cataplasma presentation quality is without significant change.

3) adjuvant addition sequence has appreciable impact with the time to mouldability and the mastic stickiness of cataplasma with forging, should be correct Select adjuvant and according to mutual relation between adjuvant and framework material and action character determine adjuvant addition sequence and control forge and Time.

11. cataplasma optimization of orthogonal test experiments

Bare substrate matched group and medicine group cataplasma compare, and ooze cloth, cohesiveness, uniformity, film residual from mastic The aspects such as amount, initial bonding strength, holding power and splitting test carry out overall merit.Redefine cataplasma formulation ingredients and ratio. Consider to affect 4 principal elements of mouldability: to sodium polyacrylate (A), dihydroxyaluminum aminoacetate (B), tartaric acid (C), polyvinylpolypyrrolidone (D) Examining or check, 3 levels of each selecting factors, arrange orthogonal test by L9 (34), factor level is shown in Table 22, and orthogonal test is shown 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 > glutinous base power, initial bonding strength is the biggest.

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

No. 3: several nothings ooze cloth, in stripping process, a small amount of matrix attachment is in lid lining, and initial bonding strength is relatively big, and cohesiveness is less, cream Containing a large amount of bubbles and a small amount of bubble cavity in body, glossiness is preferable.

No. 4: ooze cloth on a small quantity, in stripping process several without matrix attachment in lid lining, mastic uniformity is preferable, and mouldability is relatively Good, cohesiveness > initial bonding strength, containing a large amount of bubbles in mastic, glossiness is fine, fine and smooth sense.

No. 5: several nothings ooze cloth, almost without matrix attachment in lid lining in stripping process, mastic uniformity is preferable, and mouldability is relatively Good, cohesiveness > glutinous base power, initial bonding strength is less than No. 4, and bubbles volume is a lot, and glossiness is good.

No. 6: ooze cloth on a small quantity, in stripping process several without matrix attachment in lid lining, mastic uniformity is preferable, cohesiveness > just Viscous force, bubble is more, and glossiness is preferable, rich exquisiteness sense.

No. 7: several nothings ooze cloth, without matrix adhesion in lid lining in stripping process, mastic uniformity is fine, and mouldability is fine, Initial bonding strength > cohesiveness, bubble is more, and glossiness is preferable.

No. 8: several nothings ooze cloth, without matrix adhesion in lid lining in stripping process, mastic uniformity is fine, and mouldability is fine, Cohesiveness > initial bonding strength, bubble is more, and glossiness is good.

No. 9: several nothings ooze cloth, without matrix adhesion in lid lining, cohesiveness in stripping process > initial bonding strength, mastic uniformity is relatively Good, good moldability, < No. 7, bubbles volume is big, and glossiness is good for initial bonding strength.

13. initial bonding strengths, holding power and splitting measure

Cataplasma is fabricated to fixed-area 7.5cm × 11cm, every cataplasma is carried out by tester respectively the most glutinous Power, holding power and splitting measure.

14. initial bonding strength tests

Distance is set to 10cm by initial bonding strength, and bead can not be by the distance of 10cm for tangling in one minute, by its ball number As first viscous force test result.

15. holding power tests

Holding power method of testing: be attached on corrosion resistant plate by the cataplasma of 7.5cm × 11cm, with heavily anti-for 2kg rubber wheel Rolling 3 times again, steel plate, suspended from 500g counterweight, measures the time that cataplasma is completely fallen off by corrosion resistant plate.

16. splitting tests

The assay method of splitting: the cataplasma of 2.5cm × 10cm is attached on corrosion resistant plate with heavily anti-for 2kg rubber wheel Rolling 3 times again, be fixed on by steel disc on stripping tester, setting stripping pulling force is 150m/min as 200N, speed, peels off and measures Time is that cataplasma is by the time being completely exfoliated on corrosion resistant plate.

The cataplasma of different prescription proportionings is carried out instrument test, due to the cataplasma presentation quality of front 3 prescription proportionings Poor undesirable, remaining cataplasma is carried out respectively just viscous force (A), holding power (B), splitting (C) test, test result It is shown in Table 24.

Table 24 just viscous force, holding power, splitting test result

Being compared by above cataplasma integration test, No. 8 cataplasma prescription substrate not only presentation qualities are preferable, and each power Test result is of a relatively high.Skin test entirety sensation just viscous force is relatively big, and cohesiveness is relatively reduced.

17. prescription viscosifier regulation experiment groups

This experiment basis changes the ratio composition of viscosifier and cross-linking agent, continues screening catablasm base material prescription real Test, carry out overall merit from the test of the presentation quality of cataplasma, adhesion strength, glossiness and each power.Adjust adhesive agent, wetting agent Ratio with cross-linking agent.To sodium polyacrylate (A), sodium carboxymethyl cellulose (B), dihydroxyaluminum aminoacetate (C), tartaric acid (D), glycerol (E) Investigate with six factors of pH (F), experiment arrangement.Experimental group table is shown 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, stripping process remains on backing without substrate.

No. 2: several nothings ooze cloth, reflecting feel is poor, and initial bonding strength is preferable, has a large amount of bubble, and in stripping process, backing edge has A small amount of substrate residual.

No. 3: several nothings ooze cloth, bubble is less, and initial bonding strength is relatively big, remains on backing without substrate in stripping process, but light Pool degree exquisiteness sense is poor.

No. 4: oozing cloth obvious, bubble is more, remains on backing without substrate in stripping process, glossiness and fine and smooth sense are relatively Good.

No. 5: oozing cloth obvious, bubble is more, remains on backing without substrate in stripping process, glossiness and fine and smooth sense are relatively Good.

No. 6: ooze cloth on a small quantity, bubble is more, remains on backing without substrate in glass process, and glossiness exquisiteness sense is preferable.

No. 7: trace oozes cloth, a large amount of bubbles, and gloss exquisiteness sense is preferable, remains without substrate in stripping process.

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

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

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

The usage amount of cross-linking agent is different from ratio, and in manufacturing process, the viscosity of mastic is the most different, and ratio is the biggest, and it is thick Spend the least, so selecting suitable ratio, with amount, cataplasma is formed large effect.

19. initial bonding strengths, holding power and splitting test result

Cataplasma carries out just viscous force (A), holding power (B), splitting (C) test respectively, and instrument test the results are shown in Table 26.

Table 26 just viscous force, holding power, splitting test result

Measure during, the viscous force of sodium carboxymethyl cellulose is significantly less than polyvinylpolypyrrolidone, measure splitting time Time is easy to peel off, but owing to splitting is the least, no data during measuring, so the size viscous force measuring splitting should This has to a certain degree.Holding power is significantly lower than viscosifier polyvinylpolypyrrolidone group, but just viscous force is fitted in cutaneous sense is tested In, increase mutually without viscous chaeta phenomenon cohesiveness taking off patch process.

Using this prescription as the final formulation and technology of cataplasma in this is tested, later and carry out excellent on this basis Change, produce up-to-standard cataplasma product.

Embodiment 5

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

Laboratory animal: healthy adult Japan large ear rabbit, male and female do not limit, body weight 2.5～3.5Kg；Healthy adult Cavia porcellus, female Hero does not limits, body weight 250～450g；By Kunming Medical University Experimental Animal Center provide [quality certification SCXK(Yunnan) 2011- 0004.Ambient temperature 25 DEG C ± 3 DEG C, relative humidity 70% ± 15%, light application time 12h/12h.

Skin sensibiligen is tested

20 Japan large ear rabbits are selected to be randomly divided into 6 groups, often group 5.I.e. intact skins gel Matrix controls group, complete Skin compound dragon's blood gel group, intact skin compound dragon's blood gel group, broken skins gel matrix group, damaged skin compound recipe dragon Dragon's blood gel group, damaged skin compound dragon's blood gel group.Test front razor to be taken off by hair, depilation scope be 150cm2(about Quite the 10% of rabbit body surface area), check whether shaving district has damage after 24h, reject skin damage rabbit, subsequently by test medicine Uniformly it is applied to district of losing hair or feathers, and is fixed with non-stimulated gauze, sub-cage rearing；Damaged skin group first will depilation position 75% wine After essence sterilization, cause local scratch with emery cloth paper in the friction of depilation position, with oozing of blood for degree, immediately in damaged part medication, tool Body dosage and coating method are with above-mentioned intact skin group.Taking off binder after 24h, after warm water cleaning tested material, every day observes And record rabbit general manifestation and intoxication conditions, specifically include the body weight of animal, depilation district red swelling of the skin degree, hair, eye and glue The change of film, breathe, circulate, central nervous system, extremity are movable etc. change, if meeting animal dead, then postmortem in time, naked eyes Observe and carry out histopathologic examination.

Skin irritation test

Single-dose skin irritation test takes healthy Japan large ear rabbit 20, and male and female do not limit, and are randomly divided into 4 groups, i.e. Intact skin and damaged skin compound dragon's blood gel and compound dragon's blood gel group, often group 5.Test front razor to be taken off by hair Falling, depilation scope is the 10% of 150cm2(about quite rabbit body surface area).After depilation, intact skin group is by tested material primary coating On the skin of animal subject；, with after 75% alcohol disinfecting, rub at depilation position with dry sanding paper in damaged skin group first general's depilation position Wiping causes local scratch, with oozing of blood for degree, is administered at damaged part immediately, and is fixed with nonirritant gauze, after 24h, Warm water cleaning medicine, observation 1,24,48 and 72h medicine-feeding part is used to sting according to document skin with or without the situation such as erythema, edema, result Swash reaction standards of grading mark and make stimulus intensity evaluation.

Multiple dosing skin irritation test takes Japan large ear rabbit 20, is grouped and makes damaged skin ibid, continuously It is administered 7d, after each coating, fixes 12h with hospital gauze and adhesive tape, after last is administered 24h, clean medicine with warm water, observe 1, 24,48 and 72h medicine-feeding parts are marked also according to document skin wound repair standards of grading with or without the situation such as erythema, edema, result Make stimulus intensity evaluation.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 mechanism group carry out rabbit single-dose skin irritation test and show, Three, can the use of safety all without any zest.

Table 27 skin wound repair is marked

Table 28 skin irritation strength grading

The multiple dosing skin irritation table of integrals respectively organized by table 29

Skin hypersensitivity is tested

Healthy adult Cavia porcellus, male and female do not limit, and are randomly divided into 5 groups, i.e. blank group, gel-type vehicle group, compound dragon's blood gel Group, compound dragon's blood gel group and 1-CHLORO-2,4-DINITROBENZENE group (be configured to before use 1% sensitization concentration and 0.1% excite dense Degree), often group 10.Test front razor to be shaved by hair totally, scope 3cm × 3cm, after 24h, check whether shank-feathering district has damage, Reject 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-dinitro Chlorobenzene (every 0.2ml) after depilation after 24h uniform application lose hair or feathers on the right side of the back of each group of correspondence Cavia porcellus district's skin, use Gauze covers, fixing with the wrapping of non-stimulated adhesive plaster, and sub-cage rearing, remove left drug with warm water after continuing 6h.7d and 14d, is in kind repeated once.

Excite and be contacted with the 14d after last is administered sensitization, by gel-type vehicle, compound dragon's blood gel, compound dragon's blood gel With DNFB (concentration 0.1%, dosage 0.2ml) uniform application in the left dorsal depilation district skin of each group of correspondence Cavia porcellus Skin, covers with gauze, fixing with the wrapping of non-stimulated adhesive plaster, and sub-cage rearing, remove left drug with warm water after continuing 6h.At once (0h) observe dermoreaction situation, then continue to observe skin allergy situation in 24,48 and 72h.Result presses document skin Anaphylaxis standards of grading carry out scoring and sensitization evaluation, and calculate Cavia porcellus sensitivity response meansigma methods by with formula: sensitization is anti- Answer meansigma methods=(erythema forms total score+edema and forms total score)/animal number of cases；Sensitization incidence rate=occur erythema or edema (no matter Degree weight) number of animals/animal subject sum * 100%.The scoring evaluating skin allergy can be pressed by the standard of table 30 Table 31 is evaluated.

Table 30 skin allergy degree standards of grading

Table 31 hypersensitive evaluation criterion

Data statistic analysis: the data of all experimental results represent with mean ± standard deviation (x ± s), all data analysiss SPSS17.0 statistical package is carried out, carries out the comparison of many group differences with one factor analysis of variance, think during P ＜ 0.05 Difference is statistically significant.

6. sign inspection

Compound dragon's blood gel, compound dragon's blood gel, gel-type vehicle to complete, the skin conditions of damaged skin rabbit, feed, Stool, breathing, heart beating, eye and mucosa, central nervous system, extremity activity are and cause anomalous effects；Before carrying out coating, coating The weight differences of rear 3d, 7d, 14d, it has been found that compound dragon's blood gel group, compound dragon's blood gel and gel-type vehicle group matched group Relatively, there was no significant difference (table 32).In observation period 14d, there is not any acute toxic reaction in 6 groups of rabbit, dead without 1 example. Test result indicate that, compound dragon's blood gel, compound dragon's blood gel to intact skin and damaged skin skin all without the most acute Toxic reaction.

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

8. skin appearance and morphological observation

In an experiment, perusal finds, Sanguis Draxonis gel, dragon's blood gel there are no any bad shadow to normal skin Ring；To damaged skin, gel-type vehicle group before administration after 3d without significant change, start after 4d incrustation, and Sanguis Draxonis gel and Dragon's blood gel 1d after coating i.e. starts incrustation, and after 4d, incrustation starts shedding off.Experiment shows Sanguis Draxonis gel and dragon's blood gel To rabbit normal skin without any harmful effect, damaged skin is then had the effect of wound healing.

9. single-dose skin irritation test

After single smears Sanguis Draxonis gel, dragon's blood gel and gel-type vehicle, rabbit intact skin has no that erythema or edema etc. stimulate Reaction, in addition to broken skins gel Matrix controls group 2d upon administration is formed a scab, from the beginning of 4d, damaged wound all heals de- Crust.

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 that erythema or edema etc. stimulate Reaction, damaged skin group Sanguis Draxonis gel, dragon's blood gel from the beginning of 3d, damaged wound healing decrustation, again make with dry sanding paper Become oozing of blood, be administered, wound healing after 7d, return to normal condition.

11. skin hypersensitivity experiments

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

The anaphylaxis of Cavia porcellus is affected by table 33 Sanguis Draxonis gel, dragon's blood gel

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 do not limit, body weight 2.5～3.5Kg, real by Kunming Medical University Test animal center and [quality certification SCXK(Yunnan) 2011-0004] is provided.Ambient temperature 25 DEG C ± 3 DEG C, relative humidity 70% ± 15%, Light application time 12h/12h.

Animal is grouped according to " study of tcm new drug guide (pharmacy pharmacology's toxicology) " and " herbal pharmacology research method Learn " requirement, take 20 rabbit, be randomly divided into 4 groups according to body weight, often group 5.Intact skin bare substrate group, compound recipe sanguis draconis Exhaust group, compound dragon's blood group, damaged skin bare substrate group, compound dragon's blood group, compound dragon's blood group.

Medication maximum dosage, every day 1 time, continuous 2 weeks.Coating position is depilation districts, family's rabbit back spinal column both sides (being shaved off by hair with razor, depilation scope is 150cm2, about quite the 10% of rabbit body surface area), is consolidated with non-stimulated gauze Fixed, sub-cage rearing；Damaged skin group first general's depilation position, with after 75% alcohol disinfecting, is caused in the friction of depilation position with emery cloth paper Local scratch, with oozing of blood for degree, immediately in damaged part medication, concrete dosage and coating method are with above-mentioned intact skin group.

Observation index

Ordinary circumstance is observed: during administration and observe and record rabbit general manifestation and intoxication conditions every day convalescent period, has Body include the body weight of animal, depilation district red swelling of the skin degree, hair, eye and the change of mucosa, breathe, circulate, central nervous system The change of system, extremity activity etc., if meeting animal dead, then postmortem in time, perusal also carries out histopathologic examination.

Peripheral blood cell counts: be administered 2 weeks, detected when drug withdrawal next day and drug withdrawal two weeks (convalescent period), and often group takes 2 families Rabbit.Before detection, fasting can't help water about 12 hours, fixing animal, adopts entirely in fixing clear-headed Rabbit central ear artery with disposable blood taking tube Blood.Testing index includes: 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%) meter Number, leukocyte (WBC) and classification (NE%, LY%, MO%), platelet (PLT) counting, prothrombin time (PT), fibrin Former (FIB)；And bone marrow smear inspection.

Blood biochemical detects: is administered first 1st week, is administered 4 weeks and recovers 2 weeks to detect in end 24h.Animal before detection Need fasting to can't help water about 12 hours, adopt whole blood 3～6.5ml, room temperature with disposable blood taking tube in fixing clear-headed Rabbit central ear artery Lower standing about 20 minutes, is centrifuged 10min with 3000r/min, takes out serum and carries out aspartic transaminase (AST), alanine Transaminase (ALT), alkali phosphatase (ALP), gamma glutamyltransferase (γ-GGT), blood urea nitrogen (Bun), creatinine (Cre), total Albumen (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.

Systematic anatomy and histopathological examination are administered 4 weeks and terminate, in indices detect after often group randomly draw 2 move Thing is dissected, and residue animal carries out restorative observation, observes fasting after ending at indices detection and can't help water about 12h and carry out Anesthesia is put to death and is dissected, and is administered area skin and the histopathological examination of each vitals, calculates organ coefficient.

The data of all experimental results of data statistic analysis represent with mean ± standard deviation (x ± s), and all data analysiss exist Carry out on SPSS17.0 statistical package, carry out the comparison of many group differences with one factor analysis of variance, with Dunnett inspection ratio The relatively difference of mean between two groups；Think that difference is statistically significant during P ＜ 0.05.

Embodiment 7

Method of quality control is described as a example by compound dragon's blood (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1)

Peace P680 efficient liquid phase instrument worn by instrument；Wear peace chromatographic work station；Wear peace UVD170 variable-wavelenght detector；Thermo BDS HYPERSIL C18(250mm*4.6mm, 5 μm)

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 chromatograph Pure, water is heartily pure water after 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

Weighing Resina Draconis-drug 2g to be wrapped in filter paper, surname extraction, to colourless, volatilizes filter paper, and alcohol-insoluble substances content is 0.297%, it is less than 1.5% less than the alcohol-insoluble substances of States Pharmacopoeia specifications.

Sanguis Draxonis trap checks

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

Weigh 4g Sanguis Draxonis in 10ml volumetric flask, add that methanol is ultrasonic makes dissolving, methanol constant volume, after filtration, take 0.5ml Subsequent filtrate is in 10ml volumetric flask, and methanol constant volume to scale, at 284nm wavelength, trap is 3.058, more than States Pharmacopoeia specifications It is not less than 0.40.

Method of quality control is studied

(1) character

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

(2) pH value

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

(3) check

Centrifugal test takes 3 crowdes of each 10g of compound dragon's blood gel and puts in centrifuge tube, and 3000r/min takes out after being centrifuged 30min, Gel is without lamination.

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

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

The determination of content assaying method

Chromatography condition

The selection of flowing phase: investigate the flowing phase of 1% acetic acid-acetonitrile different proportion (63:37,64:36,69:31) respectively, Result is with 1% acetic acid-acetonitrile (64:36) for flowing phase, and 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one in test sample, lourerin B peak separating effect is optimal and separates Time is suitable, therefore selects 1% acetic acid-acetonitrile (64:36) for flowing phase.

The selection of detection wavelength: according to Literature Consult and the result of experiment investigation, determine that chromatographic condition is: flowing phase: 1% Acetic acid-acetonitrile (64:36), detects wavelength: 275nm, flow velocity: 1.0ml/min.Column temperature 30 DEG C.Theoretical cam curve is based on lourerin B Calculate, be not less than 5000.

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

Gel extraction method determines

Extract the selection of solvent: weigh compound dragon's blood gel 3 parts, every part of 2g, be separately added into 8ml ethyl acetate, just Butanol, methanol, it is settled to 10ml, HPLC after ultrasonic 30min and measures 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one lourerin B, result shows the separation effect of methanol Really, peak shape and content be best, so using methanol to carry out supersound extraction.

The selection of extraction time: take 2g compound dragon's blood gel 3 parts, add 8ml methanol ultrasonic 20min respectively, 30min, 40min, prepare need testing solution, and assay result shows: compound dragon's blood gel is contained by the supersound extraction time Amount has an impact, ultrasonic 30min and 40min content no significant difference, but far above 20min's.For ensureing to extract completely and contract The weakness reason time, use supersound extraction 30min.

The preparation of need testing solution

Take compound dragon's blood gel 2g in 10ml volumetric flask, add 8ml methanol, ultrasonic 30min, let cool rear methanol fixed Hold, shake up, filter, take subsequent filtrate, to obtain final product.

Specificity is tested

Removing the raw material beyond Sanguis Draxonis according to prescription, by compound dragon's blood, preparation technology makes negative control sample, presses Preparation method according to above-mentioned need testing solution prepares negative fluid, measures according to said method, result display 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B Appearance time without overlapping, shows that 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B assay are not done by substrate with negative controls appearance time Disturbing, the method specificity is good, and result is shown in Fig. 1,2,3.

Methodological study

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

Table 34 Precision test result

Reference substance is linearly investigated

Precision weighs 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 after methanol ultrasonic dissolution 25ml obtains 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one that concentration is 0.12mg/ml and concentration is the lourerin B of 0.124mg/ml.Draw 4ml, 2ml respectively, 1.5ml, 1ml, 0.5ml, 0.2ml, in 10ml volumetric flask, add methanol dilution to scale, shake up.Sample size is 20ul, injects liquid Chromatography, measures according to above-mentioned chromatographic condition, records chromatographic peak area.SPSS17.0 software data processing, with peak area (Y) For vertical coordinate, reference substance sample size (X) is abscissa, draws standard curve, calculates regression equation and 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 at 0.0004mg/ml～ In the range of 0.04mg/ml the most well, lourerin B is linear good in the range of 0.00038mg/ml～0.038mg/ml.

Replica test

Take same batch compound dragon's blood gel 6 parts, every part of 2g, prepare test sample according to the method described above, measure sanguis draconis Element A and the peak area of lourerin B, and calculate RSD value respectively 0.91%, 1.07%, show that the repeatability of the method is good, result It is shown in Table 35.

Table 35 replica test result

Sample stability is tested

Take same batch compound dragon's blood gel 2g, prepare test sample according to the method described above, by test sample at room temperature Place, respectively at the 0th, 1,2,4,6,8,12 hours, draw 20ul and inject high performance liquid chromatograph, record peak area.Result table Bright 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%, and result is shown in Table 36.

Table 36 stability test result

Sample-adding recovery test

Take compound dragon's blood gel 2g in 10ml volumetric flask, add 8ml methanol ultrasonic dissolution, be settled to after letting cool carve Degree.Taking 3 1ml subsequent filtrates after filter paper filtering in test tube, labelling sample-adding is high, medium and low.It is separately added into and 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one in sample, The 120% of lourerin B content, 100%, 80% standard substance 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one, after lourerin B 1ml fully mixes, draw 20ul and inject efficiently Chromatograph of liquid.Record peak area, measures its response rate and is all higher than 95%, and RSD% value is respectively less than 1%, shows that the method is loaded back Yield is good, the results are shown in Table 37.

Table 37 is loaded recovery test result

Sample size measures:

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

Experiment shows: compound dragon's blood gelling amount control method is: Thermo BDS HYPERSIL C18 chromatographic column (250*4.6nm, 5 μm)；Flowing phase: 1% acetic acid-acetonitrile (64:36), detects wavelength: 275nm；Flow velocity: 1.0ml/min；Column temperature 30℃.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 higher than 1.5 with this understanding, and tailing factor exists Between 0.95-1.05, theoretical cam curve is all higher than 5000, meets Pharmacopoeia of the People's Republic of China requirement.3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one exists In good linear relation in the range of 0.0004mg/ml～0.04mg/ml, regression equation is ya=136.229xa-0.21, r= 0.9999；Lourerin B is in good linear relation in the range 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%, RSD is respectively 0.10%, 0.80%。

Embodiment 8

The investigation of vitro release is described as a example by compound dragon's blood (Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1)

Release medium: three layers of gauze

Disperser and medium: using 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, and the rotating speed of magnetic stick is 200 ± 5r min-1, and dispersive medium is water-bath in advance 95% ethanol of constant temperature 37 DEG C.

Dissolution test method: take three layers of gauze, be fixed on diffusion cell, tighten, directly weighs 2g1.2% compound recipe sanguis draconis Exhausting gel on gauze, make gel uniform application on gauze with waterproof paper flicking, acceptance pool fills it up with dispersive medium, with 37 ± 0.2 DEG C of water bath heat preservation magnetic sticks, stir with the speed of 200 ± 5r min-1, respectively at 0, and 3,6,10,15,20,50, 90,120,180min samplings, take out whole liquid 8mL in acceptance pool, add the dispersive medium of equivalent 37 DEG C preheating immediately, add Time note draining bubble, acceptable solution is collected in the test tube being numbered standby.

Because the content of 3-(2,4-Dimethoxy-phenyl)-1-(4-hydroxy-phenyl)-propan-1-one and lourerin B is little in Sanguis Draxonis, directly the sanguis draconis in acceptable solution should not be surveyed with HPLC Element A and lourerin B content, use and concentrate acceptable solution, and 2ml methanol is measured after dissolving.

The process of result tries to achieve accumulative infiltration capacity according to following formula

$Q_n=\frac{\underset{i=1}{\overset{n}{\mathrm{\&Sigma;}}}{C}_i\&times;8}{2.92}$

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

Kp=Jss/C

Statistical method

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

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

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

By table 38, Fig. 6 understands, 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%, after 50min, release profiles tends to balance.After 90min, release change is little, and kept stable, lourerin B exists In 180min, average release reaches 92.94%, and about 30min release profiles tends to balance, and after 50min, release change is little, Kept stable.

Claims (3)

1. the gel containing Sanguis Draxonis compound recipe, it is characterised in that taking Sanguis Draxonis compound recipe, its compound medicine weight is 0.15g, with the mixing of 2.5mL propylene glycol, adds carbomer 9.85g, regulates with triethanolamine, and pH value is 7.0, gained gel There is the glossiness of formulation requirements, stretchability, uniformity and centrifugum；Described Sanguis Draxonis compound recipe is consisting of the dragon of weight portion Sanguis Draxonis: Borneolum Syntheticum 9:1；Or, Sanguis Draxonis: Borneolum Syntheticum 9:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.

2. the liniment containing Sanguis Draxonis compound recipe, it is characterised in that take Sanguis Draxonis compound recipe, weigh this compound medicine 0.2g is few Amount is repeatedly dissolved in propylene glycol 1.0g so that it is fully dissolve as A phase；Take 4% polyvinyl alcohol 8.0g, add carboxymethyl cellulose Sodium 0.2g and polyvinylpyrrolidone 0.5g, water-bath 70 DEG C-80 DEG C, make the abundant swelling dissolving of adjuvant as B phase；B is added to Stir in A phase mixing, is eventually adding surface active agent tween-80 and keeps the stability of liniment, and liniment is equal Even spreading upon on the glass plate scribbling liquid paraffin, 37 DEG C of drying in baking oven, the outward appearance of gained liniment has uniformity, painting Malleability, mobility and film pliability；Described Sanguis Draxonis compound recipe is consisting of the Sanguis Draxonis of weight portion: Borneolum Syntheticum 9:1；Or, Sanguis Draxonis: Borneolum Syntheticum 9:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15；Or, Sanguis Draxonis: Rhizoma Coptidis Element: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.

3. the cataplasma containing Sanguis Draxonis compound recipe, it is characterised in that take the propylene glycol-water mixing of appropriate carbomer 44:56 Solvent, swelling after obtain 20g 3% carbomer, add 50g 2% aqueous tartaric acid solution wherein, fully mix, standby；Take dragon Sanguis Draxonis compound medicine 6g, uses 15g propylene glycol: the mixed solvent of 95% ethanol 3:12,50 DEG C of water-baths are dissolved, standby；Take polypropylene Acid sodium 4g, CMC-Na 2g, aluminium hydroxide 0.3g, EDTA0.5g add in beaker, add 22g glycerol and mix well, standby；To dissolve Rear medicine adds the mixed phases such as sodium polyacrylate-glycerol, fully after mixing, is rapidly added wherein by carbomer-tartaric acid, rapidly Stir, preventing and treating local caking, it is further continued for stirring 60 minutes, treats mastic deliquescing, granular sensation is coated when disappearing, and room temperature is put After putting 3 days and get final product；Described Sanguis Draxonis compound recipe is consisting of the Sanguis Draxonis of weight portion: Borneolum Syntheticum 9:1；Or, Sanguis Draxonis: Borneolum Syntheticum 9:1； Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 9:4:1；Or, Sanguis Draxonis: berberine: Borneolum Syntheticum 1:1:0.15.