CN103007260A - Preparation method of medicine for rapidly healing burned tissue - Google Patents

Abstract

The invention relates to a preparation method of a medicine, in particular to a preparation method of a medicine for rapidly healing a burned tissue, belonging to the field of medicines. The preparation method of the medicine for rapidly healing the burned tissue comprises the following steps: (1) taking crude bromelin as a raw material; (2) taking a buffer supernatant with the pH of 6.2-7.0; (3) precipitating by using (NH4) 2SO4 with the saturation of 45%, salting out, dialyzing and desalting; (4) performing freeze-drying or precipitating a concentrated solution with cold acetone, centrifuging, and performing cryogenic natural drying; and (5) grinding and mixing refined bromelin and 10-20% of cysteine uniformly to obtain the medicine for rapidly healing the burned tissue. The decrustation time is short; for a second-degree burn, the decrustation time is generally shorter than 2 hours; clinically, the shortest decrustation time is only 30 minutes; and the decrustation is complete.

Description

A kind of for heal the rapidly preparation method of medicine of tissue of burn

Technical field

The present invention relates to a kind of preparation method of medicine, especially a kind of for heal the rapidly preparation method of medicine of tissue of burn.Belong to drug world.

Background technology

The eschar (being decrustation) of removing each burn generation is a very important link in the burn treating, and means the most commonly used remain the operation decrustation at present, but modus operandi has following deficiency: one, easily cause operation painful to patient; Its two, need standby fresh blood, in case urgent need during severe loss of blood; Its three, the situation that injures biological tissue often occurs, the particularly damage at key function position, consequence is more serious; Its four, the normal difficulty of side effect risk of anaesthetic treatment is avoided.In order to overcome the above-mentioned weak point of operation decrustation, people namely seek the medicine decrustation means since six the seventies.But medical circle is thought the decrustation medicine of having succeeded in developing, and is all not ideal enough.

The decrustation medicine comprises Chinese medicine, chemicals and biochemical enzymes preparation three major types.Decrustation Chinese medicine is commonly used has and more creates No. ten, No. one, east and ointment for treating hot water and fire scald, because of these medicines without bacteriostastis, can make traumatic infection heavy, and seldom be used.Chemistry decrustation medicine, i.e. acidic drug diffusant, acid diffusant commonly used has salicylic acid, acetone acid, succinic acid and citric acid etc., but brings the reasons such as painful because of its zest to patient, is not used clinically.The biochemical enzymes decrustation preparation is considered to more promising decrustation medicine, and it comprises again plant rennet, bacterial enzyme and animal ferment three class biochemical enzymes; But only have so far the bacterial enzyme of a kind of being called " Travase " at human Burns a small amount of application to be arranged.(" the burn treating handbook third edition, the volumes such as the poplar of side, Science and Technology of Shanghai Literature publication society in 1987; The work such as the poplar of " burn theory and practice " side, Liaoning science and technology publishing house in 1989).In plant rennet, more with bromelain (Bromelain), ficin (Dericin) and papain (Papain) research.The present invention is directly relevant with bromelain, discloses a kind of new bromelin decrustation preparation and the method for preparation thereof.

Relevant bromelain prepares the non-patent literature report of enzyme decrustation medicine first appeared in the beginning of the seventies, and patent literature then sees 74 years the earliest.

Chinese patent 91103150.2 has also been mentioned a kind of vegetable protein list preparation method, adopts the inorganic salt sedimentation method (extracting thick enzyme) and high PH roguing ion exchange (extracting smart enzyme) from Palmae (Palmaceae) Arenga (Areaga) Arenga pinnata (Areaga Pinnata, having another name called Arenga pinnata (Wurmb.) Merr.) technical process of extracting Arenga pinnata (Wurmb.) Merr. prolease in the plant sequentially comprises: adopts Arenga pinnata to do raw material → squeeze juice → ammonium sulfate and removes foreign protein, precipitation → dry left and right sides sedimentation → DE52 chromatography → lyophilization is smart enzyme preparation for thick enzyme (dialysis) is transferred PH9.5 with alkali.

The PCT patent discloses following method in the disclosed application 02825465.1 of China: by the affinitive layer purification trypsin-like serine protease of lucilia sericata (Luciliasericata) ES on aminobenzamidine agarose.The 0.025MTris-hydrochloride buffer balance columns substrate (1ml) that contains the pH8.0 of 0.5M sodium chloride with 20ml.Before upper prop, dilute natural ES (0.5ml, 70 μ g/ml albumen) with isopyknic buffer.Collection is by the fraction (0.5ml) of chromatography.After the buffer with 6.5 times of column volumes washs to remove unconjugated albumen, use free aminobenzamidine part (2ml400 μ M) to obtain the eluent of bound substances.Use fraction to collect in conjunction with the peak for analyzing with following in conjunction with (circulation) with in conjunction with the position at peak with definite at the absorbance reading of 280nm.Aminobenzamidine agarose is in conjunction with trypsin-like serine protease.After with larval enzyme secretions upper prop, unconjugated material directly by and conduct " circulation " thing (peak I) be collected.In the post buffer, add the eluent that the free amine group benzamidine obtains the protease (peak II) of combination.Do not contain the proteinase activity that is not subjected to APMSF (may comprise chymotrypsin-like enzyme) impact in conjunction with (circulation) material, and the activity in the aminobenzamidine eluting peak is eliminated (80%) by APMSF basically, the purification of prompting trypsin-like serine protease activity.The post fraction is being contained in irreducibility SDS sample buffer (the 0.5MTris-hydrochloric acid, 20% glycerol and 0.02% bromophenol blue that contain 4%SDSpH6.8) on the 12%SDS polyacrylamide gel of 0.1% human hemoglobin and checking by electrophoresis.By in 2.5%TritonX-100 (1 hour) and distilled water (15 minutes), washing to remove SDS.By overnight incubation in 37 ℃, the 0.1MTris-hydrochloride buffer of pH8.0, the proteolysis of hemoglobin substrate has produced the shown clear band of Coomassie brilliant blue protein staining in the gel in the position corresponding to protease.Each has shown the activity of several protease the fraction of starting fraction and outflow, yet has shown single band with the fraction of aminobenzamidine eluting.Therefore before the trypsin-like enzyme demonstration of identifying in the fraction with the aminobenzamidine eluting has～molecular weight of 25Kda.

The physics leaching process of one bromelain is disclosed among the CN1186118.This process comprises the pretreatment to the Fructus Ananadis comosi plant, it comprises by freezing, broken, extruding and obtains juice, filter and obtain supernatant liquid, by ultrafilter membrane and reverse osmosis membrane (reverseosmosizing? film) concentrated, and lyophilisation obtains spongiform product, etc.It is reported, can improve enzymatic activity and productive rate by control temperature, time and pH value.U.S.3658651 relates to from the stem of Fructus Ananadis comosi plant and to extract the juice that contains bromelain, juice is carried out purification by the mode of ion exchange, the anionite with bicarbonate form is used in ion exchange, the cationite that contains faintly acid functional group, and the anionite of the second bicarbonate form, then enzyme is precipitated.U.S.4,286,064 disclose the items such as preparation of bromelain crude extract.At first the juice with pineapple stem transfers to 3 or 4 with phosphoric acid with pH value, add again NaHS (Sodium? sulfhydride) oxidized to prevent sulfydryl (sulfhydryl).Inert material precipitates with for example acetone and filters.The liquid acetone precipitation of clarification, centrifugal collecting precipitation, then again dissolving and again precipitation in advance with the water that contains NaHS of phosphoric acid acidify, perhaps directly dry in vacuum tank.Crude extract can be further purified by filtration, dialysis or diafiltration (diafilatration), and to remove micromolecule and protein, the solution with gained concentrates again, last lyophilizing.For fear of degraded and/or other chemical reactions of not wanting, such as oxidation reaction, select specific treatment conditions, such as temperature, pH, solvent and buffer and/or additive, be essential such as stabilizing agent and antioxidant.Except above being not enough to, also may need crude extract is carried out further purification.Such purification can the HPLC method of example as summarizing in U.S.2002/188107.Therefore, provide bromelain pheron chemically stable and purification to need very much, it can be used to pharmaceutical composition, dermatological composition and alimentation composition, and does not have the deficiency of the preparation that contains bromelain in the prior art.Above problem is resolved by bromelain pheron or its fragment that heterogenous expression is provided, wherein said bromelain pheron or its fragment in heterologous host with soluble activity form great expression.

Though above-mentioned several patents in separation method or condition so that in the description to the decrustation enzyme difference is arranged, following some be identical: 1. used thick enzyme all extracts from pineapple stem; 2. the molecular weight of enzymatic mixture is all at 30,000-50, in 000 dalton's scope; 3. the high activity of enzymatic mixture is 35 units of every mg protein (under the cysteine existence condition); 4. isoelectric point, IP is generally all about 6; 5. the decrustation time is 4 hours the soonest.

Technical scheme

The present invention's purpose is to seek a kind of decrustation better effects if, shorter enzyme decrustation pharmaceutical preparation of time.This goal of the invention is achieved by technical scheme described below.

Of the present invention a kind of for heal the rapidly preparation method of medicine of tissue of burn, wherein:

(1) be raw material with the thick enzyme of bromelain;

(2) the buffering clear liquid with pH6.2-7.0 cleans;

(3) again with the (NH of 45% saturation ₄) ₂SO ₄Precipitate, then saltout, dialysis desalting;

(4) dry lyophilization or the concentrated solution of adopting is with cold acetone precipitation, centrifugal again low temperature natural drying;

(5) the cysteine mill that adds 10-20% by refining bromelain even the forming that be mixed.

Above-mentioned lixiviating solution precipitation employing DEAE-cellulose DE-52 carries out ion exchange and carries out drying to obtain more highly purified refining enzymatic mixture again.

When carrying out anion exchange chromatography, under 280nm, collect optical density value at 0.5 above part solution.

Also include the component of following weight portion in the above-mentioned medicine:

The acid of semi-annular jade pendant base ichthyol oil is by 5～20 parts;

2～60 parts of antibiotics or bactericides;

1～20 part of five fruit peel extract;

5～10 parts of polyhydric alcohol;

5～10 parts of sodium metabisulfite;

1～1000 part of hydrophobic oil preparation;

After above-mentioned component mixes according to weight ratio, be (0.5-2) with the weight ratio of the final products of claim 1: 1 is configured and mixes.The medicine that utilizes further proportioning of the present invention to obtain, the final effect of product are very good, and on probation through 300 patients' contrast, total effective rate and cure rate have improved respectively 7% and 4%.

Above-described ichthyol adopts Hospitals at Present or pharmacy to provide, obtain the ichthyol of national drug certification, to put down in writing like this in the medicine books: the medicine name: the acid of semi-annular jade pendant base ichthyol oil, another name: ichthyol, ichthyol, fish stone ichthyodin, classification: Western medicine, bentoquatam, the effect purposes: this product is antiseptic, gentle anticorrosion and stimulation is arranged, the effects such as antibacterial, antiinflammatory, antipruritic, secretion inhibitor thing and detumescence are arranged.Be used for furuncle and phyma, psoriasis, eczema, cervicitis, vaginitis, lymphadenitis, thrombophlebitis, chronic ulcer, chronic dermatitis, radiodermatitis, otitis externa etc.

Five fruits are to produce white semi-tropical a kind of fruit, have another name called Fructus Choerospondiatis, and the peel extract of Fructus Choerospondiatis (Choerospondiasaxillaris) is effective to the smooth healing of wound, and can pain relieving and antibacterial.

Above-mentioned polyhydric alcohol namely contains being commonly called as of polyhydroxy alcohols, can be the reduced form micromolecule sugar alcohol, also can be poly oligosaccharide or other alcohols, such as glycerol, mannitol, xylitol and the oligomeric glucose that makes with sucrose, oligofructose and oligomeric galactose etc.Add a certain proportion of some polyhydric alcohol in prescription after, proteinase activity, effectiveness can prolongedly keep.

The use of hydrophobic oil preparation can be built hydrophobic environment, forms hydrophobic outer wrapping, to enzyme and medicine particularly their solid-state form finish the film protection, allow enzyme avoid the aqueous solution state failure, also strengthened adhesion strength and permeability simultaneously.

The thick enzyme of above-mentioned albumen is bromelain, and described bromelain extraction process is as follows:

(1) fresh pineapple remove the peel, stripping and slicing, be cold depositing 10-20 hour under 5 ℃ to-5 ℃ conditions in temperature, then cool to the cold 10-20 of depositing hour pretreatment under-5 ℃ to-10 ℃ conditions, then cool under-10 ℃ to-20 ℃ conditions the cold 20-30 of depositing hour;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 10-50% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0-4.5, storage temperature is 2 ℃ to 5 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ to-35 ℃ conditions, namely obtains bromelain.

Above-mentioned bromelain extraction process is inventive point of the present invention, the product that obtains in the bromelain extraction process of the prior art can not have the physicochemical property that meets therapeutic effect well, and the inventor is through for many years improvement, with hierarchical temperature pre-cooling Fructus Ananadis comosi and membrane technology combination, the bromelain that obtains has very detailed effect in the raising of the effect of the final medicine of preparation.

Above-mentioned refining bromelain hydrolytic enzyme mixture mainly is comprised of PEPA A and PEPB B, and its molecular weight is respectively 27,000 and 30,000 dalton; The hydrolase of proteolysis of refining enzymatic mixture is 15-18 ten thousand units.

Each aminoacid of mg/100ml accounts for total amount (%) in the aminoacid composition sample of Fructus Ananadis comosi PEPA A and Fructus Ananadis comosi PEPB B

3-4 hour decrustation time, the shortest need 1 hour.

The preparation technology of bromelin decrustation preparation

(1) bromelain highly finished product technique

(1) fresh pineapple remove the peel, stripping and slicing, be cold depositing 10-20 hour under 5 ℃ to-5 ℃ conditions in temperature, then cool to the cold 10-20 of depositing hour pretreatment under-5 ℃ to-10 ℃ conditions, then cool under-10 ℃ to-20 ℃ conditions the cold 20-30 of depositing hour;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 10-50% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0-4.5, storage temperature is 2 ℃ to 5 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ to-35 ℃ conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH6.0-7.0,0.02-0.05M the phosphate buffer lixiviate, the ratio of buffer and crude protein enzyme is 15-20: 1 (V/W), 4-10 ℃ of condition lower magnetic force stirred 4,000-10 2-4 hour, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The dialysis concentrated solution that obtains is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

(2) decrustation preparation preparation

The highly finished product that make with said method add the cysteine of 10-20% and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

(3) obtain compositions Q according to following weight portion proportioning:

The acid of semi-annular jade pendant base ichthyol oil is by 5～20 parts;

2～60 parts of antibiotics or bactericides;

1～20 part of five fruit peel extract;

5～10 parts of polyhydric alcohol;

5～10 parts of sodium metabisulfite;

1～1000 part of hydrophobic oil preparation;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, with compositions Q according to (0.5-2): 1 ratio and (7) goes on foot the bromelain goods that obtain and is configured and mixes, and obtains the product of the present invention of better effects if.

(4) use the decrustation enzyme preparation to treat the method for tissue of burn

After burn patient is admitted to hospital, process as follows:

1. remove the dirt hinder on the face with Oleum Terebinthinae, clean with normal saline again and hinder face, tear off and treat that decrustation hinders face

On epidermis, such as top layer carbonization, available scalpel etc. strike off it.

2. use 1% lignocaine solution with above-mentioned decrustation enzyme dry powder furnishing pasty state, it is coated in equably hinders on the face, approximately 1-2mm.

3. watched quietly 5-10 minute, and observed and to hinder face and change, as it is many to hinder on the face little petechia, and with a large amount of sepages,

The proof decrustation is effective, needs the decrustation time short.Otherwise the decrustation time is slightly long.

4. the aseptic kpetrolatum gauze of lid layer outside the decrustation medicine again, its outer 6-8 layer sterile gauze that cover again,

Skin is fastened with binder.

5. observe the variation of wrapping gauze, the wrapping gauze is soaked into by courageous and upright sepage, proves decrustation success.Usually the shallow face of hindering only needs 2-3 hour, and the darker face time of hindering is slightly long, general 4-5 hour.It often is colourless sepage rather than courageous and upright thoroughly liquid that three degree are hindered face.

6. open wrapping, scrape off medicine and slough with tweezers, thoroughly clean wound surface with 0.5%NaHCO3 solution again.Or clean with normal saline libation at an ancient wedding ceremony gauze and to hinder face, expose the fresh face of hindering that is covered with active hemorrhage point, so far, the decrustation process is finished, and does further treatment according to different situations and processes.

(5) enzyme activity determination

Take casein as substrate, contain in the 5ml reactant liquor: 1mMEDTA; 5mM cysteine (new preparation); 1% casein, 20mM sodium phosphate buffer, pH7.0; 20-80 μ g enzyme preparation.Above solution is incubated 10 minutes at 35 ℃, add the 5ml10% trichloroacetic acid and stop enzyme reaction, filter with quantitative filter paper, measure filtrate in the optical density value of 275nm, the condition of experiment contrast and above identical, just adding first trichloroacetic acid solution before adding the substrate insulation makes enzyme deactivation (the prolease activity unit definition is: at pH7.0, the per minute caseinhydrolysate produces optical density value and equals the optical density value of 1 μ g/ml tyrosine under 275nm in the time of 35 ℃.)

The present invention compared with prior art has following good effect:

(1) adopt the thick enzyme that extracts in the pineapple fruit to make raw material, the enzyme activity of refining enzymatic mixture exceeds 5-7 doubly than prior art, and the every gram of enzyme activity is 18-19 ten thousand units;

(2) has higher yield, generally at 28-42%;

(3) the decrustation time short, second degree burn generally is less than 2 hours, only needs 30 minutes the soonest the most clinically, and decrustation is thorough.

(4) face fiber crops and use, do not find any toxic and side effects, fool proof.

(5) patient is admitted to hospital, and immediately decrustation can be removed focus as early as possible, is beneficial to hinder face and heal early.

(6) can overcome the operation misery of performing the operation when going crust and the problem of losing blood, need not anaesthetize, expense is lower, and the patient takes like a shot.

(7) be beneficial to the burn degree Accurate Diagnosis, this burn treating to the extremity functional part has profound significance.

(8) convenient transportation is preserved effect duration long, and the basic not loss of living of refrigerator cryopreservation 1 year, its enzyme was preserved 1 year under the room temperature, and its enzyme is lived and lost only is 5%.

The specific embodiment

Embodiment 1

(1) bromelain highly finished product technique

Fresh pineapple peeling, the stripping and slicing of (1) being produced from Hainan are cold depositing 10 hours under 5 ℃ of conditions in temperature, then cool to coldly under-5 ℃ of conditions to deposit pretreatment in 10 hours, then cool to cold depositing 20 hours under-10 ℃ of conditions;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 50% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0, storage temperature is 2 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ of conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH6.0, the phosphate buffer lixiviate of 0.02M, the ratio of buffer and crude protein enzyme is 20: 1 (V/W), and 4 ℃ of condition lower magnetic forces stirred 10 2 hours, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The dialysis concentrated solution that obtains is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

The highly finished product that make with said method add 20% cysteine and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

Obtain compositions Q according to following weight portion proportioning:

The acid of semi-annular jade pendant base ichthyol oil is by 5 parts;

60 parts of antibiotics or bactericides;

1 part of five fruit peel extract;

10 parts of polyhydric alcohol;

5 parts of sodium metabisulfite;

1000 parts of hydrophobic oil preparationes;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, compositions Q is configured with the bromelain goods that obtain according to 2: 1 ratio again mixes, and obtains the product of the present invention of better effects if.

Embodiment 2

(1) bromelain highly finished product technique

Fresh pineapple peeling, the stripping and slicing of (1) being produced from Guangxi, temperature for-5 ℃ of conditions under cold depositing 20 hours, then cool to and coldly under-10 ℃ of conditions deposit pretreatment in 20 hours, then cool to cold depositing 30 hours under-20 ℃ of conditions;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 10% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.5, storage temperature is 5 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ of conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH7.0, the phosphate buffer lixiviate of 0.02M, the ratio of buffer and crude protein enzyme is 15: 1 (V/W), and 4 ℃ of condition lower magnetic forces stirred 10 4 hours, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The dialysis concentrated solution that obtains is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

The highly finished product that make with said method add 15% cysteine and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

(3) obtain compositions Q according to following weight portion proportioning:

The acid of semi-annular jade pendant base ichthyol oil is by 20 parts;

60 parts of antibiotics or bactericides;

20 parts of five fruit peel extracts;

10 parts of polyhydric alcohol;

10 parts of sodium metabisulfite;

1000 parts of hydrophobic oil preparationes;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, compositions Q is configured with the bromelain goods that obtain according to 1.5: 1 ratio again mixes, and obtains the product of the present invention of better effects if.

Embodiment 3

(1) bromelain highly finished product technique

(1) Guangxi fresh pineapple peeling, stripping and slicing, temperature for-2 ℃ of conditions under cold depositing 15 hours, then cool to and coldly under-7 ℃ of conditions deposit pretreatment in 15 hours, then cool to cold depositing 25 hours under-15 ℃ of conditions;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 30% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.2, storage temperature is 3 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-35 ℃ of conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH6.5, the phosphate buffer lixiviate of 0.03M, the ratio of buffer and crude protein enzyme is 18: 1 (V/W), and 7 ℃ of condition lower magnetic forces stirred 63 hours, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The poly-dialysis concentrated solution that 3. obtains of step is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

The highly finished product that make with said method add 12% cysteine and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

Obtain compositions Q according to following weight portion proportioning:

The acid of semi-annular jade pendant base ichthyol oil is by 20 parts;

2 parts of antibiotics or bactericides;

20 parts of five fruit peel extracts;

5 parts of polyhydric alcohol;

10 parts of sodium metabisulfite;

1 part of hydrophobic oil preparation;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, compositions Q is configured with the bromelain goods that obtain according to 2: 1 ratio again mixes, and obtains the product of the present invention of better effects if.

Embodiment 4

(1) bromelain highly finished product technique

(1) the fresh pineapple in Hainan peeling, stripping and slicing, temperature for-5 ℃ of conditions under cold depositing 15 hours, then cool to and coldly under-8 ℃ of conditions deposit pretreatment in 15 hours, then cool to cold depositing 25 hours under-15 ℃ of conditions;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 40% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0, storage temperature is 2 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-35 ℃ of conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH6.5, the phosphate buffer lixiviate of 0.04M, the ratio of buffer and crude protein enzyme is 19: 1 (V/W), and 8 ℃ of condition lower magnetic forces stirred 9 2.5 hours, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The dialysis concentrated solution that obtains is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

The highly finished product that make with said method add 10% cysteine and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

The acid of semi-annular jade pendant base ichthyol oil is by 5 parts;

2 parts of antibiotics or bactericides;

1 part of five fruit peel extract;

5 parts of polyhydric alcohol;

5 parts of sodium metabisulfite;

1 part of hydrophobic oil preparation;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, compositions Q is configured with the bromelain goods that obtain according to 0.5: 1 ratio again mixes, and obtains the product of the present invention of better effects if.

Embodiment 5

(1) bromelain highly finished product technique

(1) Guangxi fresh pineapple peeling, stripping and slicing, is cold depositing 10-20 hour under 5 ℃ to-5 ℃ conditions in temperature, then cools to the cold 10-20 of depositing hour pretreatment under-5 ℃ to-10 ℃ conditions, then cool under-10 ℃ to-20 ℃ conditions the cold 20-30 of depositing hour;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 10-50% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0-4.5, storage temperature is 2 ℃ to 5 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ to-35 ℃ conditions, namely obtains bromelain.

(4) get a certain amount of Fruit Bromelain semifinished product, use pH6.0-7.0,0.02-0.05M the phosphate buffer lixiviate, the ratio of buffer and crude protein enzyme is 15-20: 1 (V/W), 4-10 ℃ of condition lower magnetic force stirred 4,000-10 2-4 hour, 000 rev/min centrifugal about 10 minutes, collect supernatant; Or remove insoluble impurity in the lixiviating solution with Filtration;

(5) collection liquid precipitates with the ammonium sulfate of 45% saturation, and precipitation process is finished very soon; The centrifugal supernatant that removes;

(6) precipitate is used distill water dialysis desalination (1: 100) again, sloughs ammonium sulfate, gets concentrated solution;

(7) the concentrated solution lyophilizing gets final product to get the bromelain highly finished product, yield 20-34% (because of thick enzyme source difference).The poly-dialysis concentrated solution that 3. obtains of step is available cold acetone precipitation also, and apparatus lid centrifuge tube refrigerated centrifuger is centrifugal 10 minutes again, and the precipitate natural low temperature is dry, can obtain purer proteolytic enzyme mixt dry powder.The bromelain hydrolytic enzyme highly finished product that said method obtains, hydrolytic enzyme vigor are every gram 15-18 ten thousand units; Protein content 16-20%.

If need the more highly purified mixed enzyme goods of preparation, then lixiviating solution can be carried out ion-exchange chromatography with DEAE-cellulose DE-52, collect two absworption peaks that have proteinase activity at the 280nm place, collect the part of optical density value more than 0.5.Collection liquid carries out lyophilization and can obtain the higher highly finished product of purity.

(2) decrustation preparation preparation

The highly finished product that make with said method add the cysteine of 10-20% and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product, low temperature is moistureproof preserve stand-by.

(3) obtain compositions Q according to following weight portion proportioning:

The acid of semi-annular jade pendant base ichthyol oil is by 5～20 parts;

2～60 parts of antibiotics or bactericides;

1～20 part of five fruit peel extract;

5～10 parts of polyhydric alcohol;

5～10 parts of sodium metabisulfite;

1～1000 part of hydrophobic oil preparation;

Above-mentioned component obtains compositions Q after mixing according to weight ratio, with compositions Q according to (0.5-2): 1 ratio and (7) goes on foot the bromelain goods that obtain and is configured and mixes, and obtains the product of the present invention of better effects if.

Through using, in the 1st hour scope in by 2 days, all visible skin corium cutaneous appendage, body of gland, hair follicle regeneration is abundant; The inventor has compared the hair follicle number of the burn wound of blank group and product of the present invention, 0～10 of the every low-power field of the newborn epidermis hair follicle number of blank group, several 22～44 of the every low-power field hair follicle of product treatment group of the present invention, P＜0.01.This results suggest product of the present invention uses and can promote the appendicular regeneration of burn wound's skin histology and reparation.The wound surface collagen of group of products of the present invention is arranged near normal skin, and blank group (d) wound surface collagen is arranged unordered, and the scarring tendency is arranged, and presents " singleization " vitreous degeneration, lacks the cutaneous appendages such as sweat gland.This results suggest product of the present invention uses promoting the orderly rearrangement of wound surface collagen and the differentiation of skin affiliated group that obvious effect is arranged.Downright bad exfoliation behind the burn, newborn epithelial tissue is along the covering of creeping of leukocyte infiltration band.Show that clearly the rear epidermis cell of burn is from the epithelization process of deep skin differentiation.This results suggest, the burn model that this experiment is adopted can know that demonstration burn slough comes off, the dynamic process in each period that the differentiation of newborn epidermal tissue is creeped.The quantitative analysis that is conducive to test and comparison, and observe recovery and the regeneration of damaged tissues in different time sections, by detailed pathological observation, can the impaired wound inflammation of clear demonstration ooze out (impaired wound surface does not excise, and this is one of this model beneficial effect of being different from holostrome skin excision model).

The immunohistochemical staining result, the brown dense positive particle that dyes of group of products of the present invention is deposited on epidermis cell, fibroblast and the body of gland, and blank group positive expression rate is starkly lower than group of products of the present invention (absorbance), (P＜0.05).The inventor uses the secretion that the basic fibroblast growth factor in the wound tissue (FGF) and epithelical cell growth factor (EGF) have detected in streptomycin peroxidase complex immunity histochemical method.Discovery is repaired the expression of phase group of products of the present invention on blood vessel and body of gland at inflammation edema phase and collagen and is obviously increased to some extent (than blank), but FGF does not continue the trend (FGF continues high expressed has scarring to form trend) that increases.Point out group of products growth factor expression of the present invention that wound repair is had good regulating action.

Model case

The treatment burn surface area is at 500cm ²Above, burn 1 degree 66 people, burn 2 degree 78 people, cure rate 98%, effective percentage 99.5% is all without cicatrix.

The treatment burn surface area is at 20c m ²Above, burn 1 degree 44 people, burn 2 degree 52 people, cure rate 98%, effective percentage 99.5% is all without cicatrix.

The clinical practice statistics

Lee, man, people from Hubei, the Calx of in one action, accidentally stepping on, foot was burned, with ice compress more than 1 hour, get blister subsequently the long 10cm of wound, wide 2cm, with the Scald Ointment of company's infirmary, worsened redness afterwards in three days, flow densely, Drug therapy of the present invention was used in a burst of twinge on the 4th day, take a turn for the better two days later decrustation recovery from illness after the week.

Lee, the woman, people from Qinhuangdao, the burn that catches fire, head and shoulder burn surface area reach 600cm ², have no improvement in Tangshan City's hospitalize a couple of days, with medicine treatment of the present invention, only be clearly better with 3 days burn site, be used in conjunction this medicine and substantially fully recovered in 9 days, without cicatrix.

Pay certain, the woman lives people from Hubei, and long-term sick Lying bed does not rise, and boiled water is scalded, and area is very large, has two places to reach 20cm ²Above, with this medicine treatment recovery from illness in 15 days.

Permitted unity and coherence in writing, man, 3 years old.Because boiled water is scalded right lower limb, area 4%, is mixed, went to a doctor in rear 1 hour in scalding, covers plaster contg. tuber of hyacinth bletilla for treating burns through debridement with II degree, III degree, behind the enclosing cover sterile gauze, with bandaging, pain is namely ended, the 1st day, changed dressings in 12 hours 1 time, thereafter change dressings every day 1 time, stop to oozing out, change dressings every day 1 time, anti symptom treatment was fully recovered in 12 days simultaneously.

Liu Junxiang, woman, 48 years old.Facial area and cervical region boiled water are scalded, area 3%, the II degree is scalded, and goes to a doctor Dual-eyelid height swelling during first visit in rear 24 hours in burn, the vesicle that as seen facial area and cervical region differ in size, make a definite diagnosis eyeball section without after scalding, after debridement, cover plaster contg. tuber of hyacinth bletilla for treating burns, the eye anti symptom treatment, change dressings recovery from illness on the 4th every day 1 time.

In beautiful, woman, 62 years old.Two buttocks, thigh boiled water scald, II degree when having a bath, area 6%, medical in hindering rear 36 hours, two buttocks, the swelling of thigh and calf height during first visit, as seen the vesicle that differs in size, as seen low-grade infection covers plaster contg. tuber of hyacinth bletilla for treating burns, through anti symptom treatment such as anti-inflammations behind debridement, disinfection, change dressings recovery from illness in 8 days every day 1 time.

Li Yujin, people from Hunan, man, 4 years old.The right leg boiled water is scalded, area 2%, and II degree burn was gone to a doctor in scalding, is covered plaster contg. tuber of hyacinth bletilla for treating burns through debridement in rear 1 hour, and behind the enclosing cover sterile gauze, with bandaging, pain is namely ended, and children's's sob stops.The 1st day, changed dressings 1 time in 12 hours, changed dressings 1 time in per two days thereafter, stop to oozing out, change dressings every day 1 time, anti symptom treatment was fully recovered in 7 days simultaneously.

Zhang Ru, people from Shandong, woman, 33 years old.The I degree, 5% scalds right lower limb, covers plaster contg. tuber of hyacinth bletilla for treating burns through debridement because of boiled water in rear 4 hours of burn is medical, and behind the enclosing cover sterile gauze, with bandaging, pain is namely ended.The 1st day, changed dressings 1 time in 12 hours, change dressings thereafter recovery from illness in 3 days every day 1 time.

Song Yuping, woman, 41 years old.II, III degree, breast, abdomen, lap, area 9%, medical in rear 24 hours of burn, breast during first visit.Abdomen.The vesicle that as seen lap differs in size and one's water breaks bleb skin, the surface forms shallow in ulcer, covers plaster contg. tuber of hyacinth bletilla for treating burns after debridement, covers with sterile gauze, changes dressings recovery from illness in 19 days every day 1 time.

Liu Zhongzhi, man, 56 years old.In rear 24 hours of burn medical scald right lower limb, area 4%, II degree because of boiled water and scald, in scald rear 1 hour medical, cover plaster contg. tuber of hyacinth bletilla for treating burns through debridement, behind the enclosing cover sterile gauze, with bandaging, pain is namely ended.The 1st day, changed dressings 1 time in 12 hours, change dressings every day thereafter 1 time, stop to oozing out, change dressings thereafter recovery from illness in 12 days every day 1 time.

Yang Xia, woman, 25.People from Hubei, the thigh boiled water is scalded, area 2%, and the II degree is scalded, and scalds right lower limb, covers plaster contg. tuber of hyacinth bletilla for treating burns through debridement, behind the enclosing cover sterile gauze, with bandaging because of boiled water in rear 24 hours of burn is medical.The 1st day, changed dressings 1 time in 12 hours, stop to oozing out, change dressings every day 1 time, anti symptom treatment was fully recovered in 5 days simultaneously.

Liu Rennan, the Pekinese, boiled water is scalded right lower limb, area 4%, II degree, went to a doctor in rear 1 hour in scalding, is covered plaster contg. tuber of hyacinth bletilla for treating burns through debridement, and behind the enclosing cover sterile gauze, with bandaging, pain is namely ended, and children's's sob stops.The 1st day, changed dressings 1 time in 12 hours, change dressings every day thereafter 1 time, stop to oozing out, every Mantidis is changed dressings 1 time, and anti symptom treatment was fully recovered in 12 days simultaneously.

Liu, man, soldier, 24 years old.The facial area electric welding II degree of burning, medical in rear 2 hours of burn, Dual-eyelid height swelling during first visit, the vesicle that as seen facial area and cervical region differ in size covers plaster contg. tuber of hyacinth bletilla for treating burns after debridement, and the eye anti symptom treatment is changed dressings 1 every day, recovery from illness on the 4th.

Yang Xiaopeng: the male, the Pekinese, 19 years old, the right upper arm deep second degree burn, area 1.3% is scalded and was admitted to hospital in rear 27 hours.Conventional cleaning is hindered face, occurs little petechia after 5 minutes with decrustation preparation coating of the present invention, wraps up after 4 hours, and slough is softening, strikes off slough, cleans and hinders face, and decrustation is very thorough, hinders the face healing after routine was changed dressings 9 days.

Shujuan WANG: the women, because the lonely light of the electricity chemical fibre clothes that ignites causes facial and two burn of upper limb, the gross area 10%, two upper limb are third degree burn, area 6% is hindered rear 10 hours with decrustation preparation decrustation of the present invention.Hindered face in 6 minutes after coating this product and little petechia occurs, after 3 hours 40 minutes, fully decrustation is removed slough and is cleaned and hinders face, hinder the face substrate and still be corium, cover with the radiation Corii Sus domestica and hinder face, after 21 days, hinder face and substantially heal, without scar hyperplasia, the both hands function is normal when leaving hospital in 41 days in hospital.

Claims (7)

1. one kind is used for heal the rapidly preparation method of medicine of tissue of burn, it is characterized in that:

(1) be raw material with the thick enzyme of bromelain;

(2) the buffering clear liquid with pH6.2-7.0 cleans;

(3) again with the (NH of 45% saturation ₄) ₂SO ₄Precipitate, then saltout, dialysis desalting;

(4) dry lyophilization or the concentrated solution of adopting is with cold acetone precipitation, centrifugal again low temperature natural drying;

(5) the cysteine mill that adds 10-20% by the refining bromelain even product of the present invention that forms that is mixed.

2. method according to claim 1 is characterized in that: described lixiviating solution precipitation adopts DEAE-cellulose DE-52 to carry out ion exchange to carry out drying with the more highly purified refining enzymatic mixture of acquisition again.

3. method according to claim 2 is characterized in that: when carrying out anion exchange chromatography, collect optical density value at 0.5 above part solution under 280nm.

4. method according to claim 1 is characterized in that: described highly finished product add the cysteine of 10-20% and make activator, and mill is mixed even, and packing is decrustation preparation dry powder finished product.

5. method according to claim 1 is characterized in that: the component that also includes following weight portion in the described medicine:

The acid of semi-annular jade pendant base ichthyol oil is by 5～20 parts;

2～60 parts of antibiotics or bactericides;

1～20 part of five fruit peel extract;

5～10 parts of polyhydric alcohol;

5～10 parts of sodium metabisulfite;

1～1000 part of hydrophobic oil preparation;

After above-mentioned component mixes according to weight ratio, with the weight ratio of the final products of claim 1 be (0.5-2): 1.

6. such as the described method of claim 1-3, it is characterized in that: the thick enzyme of wherein said albumen is bromelain, and described bromelain extraction process is as follows:

(1) fresh pineapple remove the peel, stripping and slicing, be cold depositing 10-20 hour under 5 ℃ to-5 ℃ conditions in temperature, then cool to the cold 10-20 of depositing hour pretreatment under-5 ℃ to-10 ℃ conditions, then cool under-10 ℃ to-20 ℃ conditions the cold 20-30 of depositing hour;

(2) raw material that thaws of disintegrating machine quick crashing, in shattering process, the water that can add Fructus Ananadis comosi gross weight 10-50% weight portion, carrying out scum juice at squeezer separates, the pH value of juice is adjusted to 4.0-4.5, storage temperature is 2 ℃ to 5 ℃, and squeezing juice removes by centrifugalize and filter pressing and residually look into, easy flocking settling thing, obtains clear liquid;

(3) clear liquid adopts the membrane technology purification concentrated, hold back the material of 20,000 above molecular weight with ultrafiltration UF equipment, see through water and micromolecule, carry out the reverse osmosis molecular cut off more than 1000 with reverse osmosis RO equipment again, obtain high-concentration enzyme liquid, high-concentration enzyme liquid is iced to subzero solidifying, at dry vacuum container the enzyme piece drying of solidifying is become sponge shape enzyme product, high-concentration enzyme liquid preferably is positioned over anxious congealing admittedly under-25 ℃ to-35 ℃ conditions, namely obtains bromelain.

7. method according to claim 1 is characterized in that:

Described refining bromelain hydrolytic enzyme mixture mainly is comprised of PEPA A and PEPB B, and its molecular weight is respectively 27,000 and 30,000 dalton; The hydrolase of proteolysis of refining enzymatic mixture is 15-18 ten thousand units.