DE4000797A1 - BIOLOGICAL ACTIVE MATERIAL, METHOD FOR THE PRODUCTION THEREOF, AND CONCENTRATIVE AGENT - Google Patents

Description

The present invention relates to a biologically active Material, a process for its production and an on its foundation produced dressing means.

The biologically active material is used in the treatment of trauma, frostbite, trophic ulcers, purulent and necrotic wounds and in diseases accompanied by the formation of necrotic sections and the accumulation of a dense and viscous exudate in surgery, stomatology, urology, Proctology and gastroenterology. In addition, the biologically active material in veterinary medicine and in biotechnology, for. B. as a sorbent in the preparation of trypsin inhibitors - α _I -Antitrypsin - are applied.

In medicine, native proteolytic enzyme preparations for the treatment of purulent and inflammatory diseases known. However, these enzymes are expensive and difficult to obtain. Although these enzymes have a certain effect they are unstable to those in the  Wound secretion contained inhibitors as well as against pH Changes and temperature factors. Also available the said enzymes have antigenic (allergic) and pyrogenic properties.

Known is a film-like biologically active substance containing a carrier, for. Triacetate and secondary acetate of cellulose, as well as a proteolytic ferment, e.g. Trypsin, α-chymotrypsin. The components mentioned are composed as follows: 80-99% by weight of triacetate (secondary acetate) of the cellulose and residual ferment (SU-PS 7 00 138).

This material is made by emulsifying an aqueous solution of a proteolytic enzyme in cellulose ester solution, in organic solvent that is immiscible with water is recovered, with subsequent formation of the film. The proteolytic ferment is in the finished material in shape smallest inclusions available.

This material has the ability through the constant Secrete small enzyme doses from its surface long necrolytic effect.

After the exit of the ferment from the material it receives however, the properties of a native ferment with all its disadvantages such as instability to inhibitors, pH and temperature changes as well as antigenicity and Pyrogenicity. In addition, be on a considerable Consumption of the ferment of up to 20% by mass noted.

The high probability of inactivation of enzymes in organic solvents used for the production of Foil material or monofilaments are used, meaning limits the use of this material  Material as such can only be used as drainage material be, as a dressing material, it is not suitable.

Also known is a biologically active granule material, contains the carrier substances, for. B. compounds of the type Dextran, chitin, chitosan and those immobilized thereon Ferments chymotrypsin and trypsin (FR-OS 25 56 222). you are composed as follows: 80.0-80.5% by mass Vehicle, residual ferment. The material has a prolon greedy effect. The ferment has no way in to enter the circulation; it is not antigenic.

Among the shortcomings of the material include the increased Content of ferment (up to 20% by mass) and the high cost. In addition, the storage of the material in extra zube rode buffer solutions at low temperatures (up to 2 ° C) requires special equipment and equipment.

A method for producing a biological is known active material, which is carried out as follows:

• - It provides an aqueous solution of a proteolytic Ferments, preferably from trypsin, forth;
• - In a water-immiscible organic solution Medium provides a solution of the cellulose ester, in this case of cellulose triacetate, forth;
• the first solution is emulsified in the second;
• - The dry process turns the material into medicines shapes, z. As platelets, films, etc., manufactured;
• - The finished pharmaceutical forms are at 25 ° C. held in a vacuum drying oven for one hour (SU-PS 7 00 138).

After this process became biologically active materials obtained, which are composed as follows:

Cellulose triacetate or secondary acetate of cellulose 80 to 99 mass% trypsin 1 to 20% by mass

The disadvantages of this method are the following:

• - high enzyme consumption (up to 20%),
• - The enzyme loses its activity when emulsified in one partially organic solvent,
• - there is a lack of chemical bonding between the ferment and the vehicle, which is due to the fact that the ferment only with the help of physical forces in the Material is included,
• - allocation of enzyme in wound exudate in the form of a native drug can lead to negative side effects, z. B. lead to allergic reactions, and
• - it is a special equipment, a vacuum dry cupboard, required.

The obtained by this method in the form of platelets Material can not be used as bandage because it does not have the necessary sanitary hygienic properties such as hygroscopicity, air permeability, elasti and so on. The material can only as Dräna serve.

A further method for producing biolo is known gisch active substances, in which one particles of the material (the vehicle) - crosslinked dextran, chitin and Chitosan - allowed to swell in water. It will activate one dissolution of 2-amino-4,6-dichloro-sym-triazine in acetone prepared, which is diluted at 50 ° C with water. The Material is stirred at 50 ° C with the activating Solution offset. To the activating solution, in which the  Material is an aqueous Natriumcarbonatlö solution and hydrochloric acid. The solution is reacted at 50 ° C stir. The resulting mixture is filtered. The stay Bende material is treated with an aqueous acetone solution and Water rinsed. This gives the activated material. which can be stored at 2 ° C in a phosphate buffer. It is a fermentation solution, for. A chymotrypsin buffer solution produced. The ferment is activated on the Material - Crosslinked Dextran, Chitin and Chitosan - Immobi lisiert. The material with immobilized chymotrypsin is alternately in a borate and acetate buffer solution rinsed the sodium chloride in different concentra tion contains. The material is sodium chloride-free Rinsed borate buffer. The material with the immobilized Ferment is lyophilized (FR-OS 25 56 222, DE-OS 34 44 746, CS-PS 2 49 311).

After this process became biologically active materials obtained, which are composed as follows:

ferment 19.5 to 20% by mass vehicle rest

This method has the following major disadvantages:

• - high fermentation consumption (up to 20%),
• - complex manufacturing technology and therefore one low productivity and high costs,
• - rare and exotic raw materials, eg. Chitin from crab armor or from the mycelium of Penicillium chrysogenum, used,
• - There are special equipment for storage and lyophilis required for the material.

The biologically active Material can not be used as bandage, but only as Scattering powder or suspension used because it as  End product a powder with immobilized ferment with a Particle size of 0.01-0.5 mm represents.

A biologically active bandage is known which is composed of three layers, a protective layer, an absorbent layer and a wound contact layer (GB-PS 20 92 006). This remedy has a thin ferment layer, which is on the inside of the wound contact layer is located. The protective layer consists of either a woven or non-woven fabric.

The absorption layer consists of cotton or viscose (Cotton wool) and artificial silk.

The wound contact layer is made of woven or non-woven Textiles or of a polyester, polyamide, polyure than, polyethylene, polypropylene or cellulose triacetate produced hole film and contains metallic copper or Copper compounds.

In the interaction between the wound secretion and the Each bandage agent all have negative characteristics of the ferment in native form. There are this specifically the instability to the inhibitors, which are contained in the wound secretion against pH change and temperature fluctuations, antigenicity and pyroge quality as well as the high costs caused by the increased fer ment consumption are caused.

The aim of the present invention is to provide such a biolo gisch active material that the effect of in Pharmaceuticals prolonged for him Storage in packaged condition under normal conditions while maintaining all sanitary hygiene properties a dressing material, in which the healing effect  faster and the content of biologically active substances it is lower and effective as a bandage can be used.

Another object of the invention is the development of a such process for the production of a biologically active Materials through which the substance obtained as Verbandmateri al, which is technologically simple, none due to the use of accessible raw materials requires a great deal of material and material industrially easily manufactured.

Another object of the invention is a dressing means to create, with significantly lower fermentation consumption has a high healing effect, due to stability against the inhibitors in the wound secretion and against pH Changes and temperature fluctuations are distinctive, as well has no antigenic and pyrogenic properties.

This object is achieved in that a new biologically active material was developed that a carrier and an enzyme immobilized thereon contains, according to the invention as a carrier, a fabric is used. It is immobili in the material on him fermented fermentation with the textile carrier in covalently linked to the following composition:

ferment 0.02 to 0.50 mass% carrier 99.98 to 99.50% by mass

The fabric used in the biologically active material is z. B. a non-woven or a woven or knitted fabric The substance on which the fermentation is immobilized.

The fabric is used because it itself has a weak healing effect; he drains the  Wound, absorbs microbes and toxins, protects the Oberflä It protects the wound from dirt and prevents too much Dehydration.

The fabric can be made of natural fibers, eg. B. off Cotton, flax, silk and wool fibers, or syn thetic fibers, e.g. B. polyacrylonitrile, polycaproamide and polyurethane fibers. It is suitable also synthetic fibers, z. B. triacetate fibers from secondary Cellulose acetate or viscose fibers.

According to the invention that is immobilized on the fabric Ferment with this substance in the following composition of mentioned components of the material according to the invention covalently linked:

ferment 0.02 to 0.50 mass% fabric 99.98 to 99.50% by mass

This ratio is due to the fact that the use of the biologically active material having a fermentation content less than 0.02% by weight drastically reduces the healing effect reduces (by more than 1.5 times). The use of the Material with a fermentation content of more than 0,50 mass% shortens the treatment time, which averages 14 ± 2 days is not essential. The cost of the material and the However, treatment will be much higher in this case.

Under the conditions of medical treatment be contributes the duration of contact of the material according to the invention as with the wound expediently 48 to 96 hours.

When stored under normal conditions (room temperature of about 20 ° C in packaged form), the material remains minde effective for at least five years.  

By applying the material can according to the invention of Healing effect on average by 1.5 times at 1/10 to 1/30 of the conventional doses compared to the recovery time with the help of known biologically active materials or accelerated native ferments. The invention Material is non-toxic and does not cause allergic Reactions.

According to the invention, the textile carrier is expediently made made of natural fibers. That ensures better Contact conditions between the wound and the material that very close to the comfort conditions.

To expand the raw material base, it is according to the invention expedient, the textile carrier of synthetic or Synthetic fibers produce.

According to the invention it is also expedient that the material proteolytic (hygrolytin, protosubtilin, terrilytin, Trypsin), collagenolytic (collagenase) or bacterioly contains (lysozyme) ferments through which denatured Proteins of different structure can be decomposed and that contribute to wound cleansing. The invention Material produced may vary depending on the shape of the wearer Towels, bandages, bandages, etc. be executed.

According to the invention it is also appropriate that the biological active material a Scharpie with 1.0 to 15 mm fibers represents, making this material in stomatology and for treatment deeper, z. As purulent puncture wounds compli ornamented form, can be used.

The material according to the invention can also be used in For Powder with a particle size of 0.001 to 1.00 mm available. In this case, the material is suitable for  Treatment of large wound surfaces and wound surfaces compli decorated configuration.

The material is suitable for treating purulent and necrosis wounds, acute and chronic periodontitis, to Treatment of wounds complicated configuration, of Cavities and hollow organs of the living organism, the have a narrow exit channel, such. For treatment of inflammation in the urinary bladder.

In the experimental testing of the invention Material and during clinical investigations found that it is biologically and therapeutically superior is active. Special studies have shown that it in the first four days after contact with the wound is particularly active.

The animals tested were not allergic Reactions to the biological obtained according to the invention active material detected. Changes in functions Hemostasis as the content of products of fibrinogen decay, as well as the fibrinogen content and the fibrinolyti have not been detected.

In the investigations of bacterial cultures from the wounds before and after the application of the material according to the invention could narrow the spectrum of applied microor organisms, a decrease in their virulence due to Replacement by a less pathogenic flora and a clot resistant to antibiotics. The application of the material according to the invention leads to a more intensive wound surface sterilization, to a Reduction of preparation time from sick to plasti operations and to shorten the inpatient medical treatment of patients by 10 Days.  

During the entire treatment period were urinary and Blood analyzes of the patients made and their temperature measured. By the application of the material according to the invention as induced general or local complications were never registered.

The material according to the invention can be described, for example, as Ver be used, the one of at least three Layers represents existing association, a protection layer, an absorption layer and wound contact layer.

The bandage can be produced in several variants be, for. B. with one or two layers of the biological active textile material and the covalently bonded with him immobilized enzyme.

The protective layer of the dressing means is according to the invention in all variants of woven, non-woven or ge knitted fabrics made.

The wound contact layer according to the invention in all Her variant of the dressing means from a biological active material produced, which is a textile carrier from natural or synthetic or synthetic fibers and an on collagenolytic (collagen nose), proteolytic (Hygrolytin, Protosubtilin, Terrily tin, trypsin) or bacteriolytic (lysozyme) ferment contains. Here is the relevant ferment with this Carrier in the following composition covalently in mass% connected:

Carrier | 99.98 to 99.50 ferment 0.02 to 0.50

The textile material for producing the Wundkontakschicht the dressing means represents a knitted, woven or nonwoven fabric.

In one embodiment of the bandage means with a Layer of the biologically active material as a contact layer The absorption layer is made of a woven, non-woven or knitted fabric without a biologically active substance manufactured.

In a further embodiment of the bandage means with two layers of biologically active material one the absorption layer of a material ago, the one bacteriolytic ferment, e.g. B. lysozyme. This Material is a woven, nonwoven or knitted one Material. The ratio between ferment and carrier is in mass% 0.02-0.50: 99.98-99.50. In addition, the Absorption layer as a weft (cotton wool) from a Textilma material with an immobilized bacteriolytic ferment be prepared, the fiber length of 1.0 to 15 mm is, or from a powder of the same material with a granule size of 0.001 to 1 mm.

By elements of the dressing material such as adhesive plaster, Velcro band etc. it can be attached to the patient.

The bandage is made according to traditional technologies of Textile and garment industry produced.

By the application of the dressing means according to the invention a few hours after the onset of the trauma (after the Injury) can prevent infection and the following treatment can be shortened by two to four days.

The biologically active material described above becomes  produced according to the invention in two stages: by activity tion of a chemically inactive carrier, in this case Textile carrier in the form of fabrics, and subsequent Immobilization of the biologically active substance on the activated carrier, in this case a ferment, z. B. Protosubtilin, hygrolytin, terrilytin, trypsin, lysozyme and Collagenase.

Activation is the introduction reactive functional groups in the textile carrier. The activation process depends on the chemical composition tion (natural, synthetic and artificial fibers) and of the Textile structure of the support (braid structure and thickness and yarn speed, density of the fabric, etc.). The Activation is up to a content of reactive functional groups from 0.0625 to 3.125 mg eq. per gram carried out by the wearer.

A cellulosic textile carrier is characterized by its oxida tion with alkali periodate, z. As sodium periodate activated until dialdehyde cellulose is formed. The activated textile carrier ger thus contains 0.0625 to 3.125 mg eq. reactive functional groups per gram of carrier.

A textile carrier made of synthetic fibers (carrier without Cellulose), e.g. B. from polycaproamide fibers, is acid hydrolysis of the carrier with subsequent rinsing and by Reaction of the hydrolyzed carrier with a solution akti Fourth, a substance with reactive functional Contains groups. After the content of these groups in the Carrier 0.0625 to 0.3125 mg eq. per gram of vehicle he is enough, the activation is terminated. Is appropriate the use of strong inorganic acids, eg. Hydrochloric acid or sulfuric acid. The hydrolyzed carrier of syntheti fibers should either be treated with a glutaraldehyde solution  followed by rinsing with water or with a dime thylsulfatlösung with subsequent rinsing of the carrier with Ethyl alcohol and processed with phosphate buffer.

The immobilization of the biologically active substance, in In this case of ferment, on the activated textile carrier takes place on a buffer solution with a pH of 6.5 to 7.5 at room temperature over 8 to 16 hours. there arise due to the reaction of the reactive groups of the textile carrier with the free functional groups of the biologically active substance, in this case ferments, Covalent chemical bonds between the textile carrier and the ferment. As a result, the textile material with a chemically (covalently) immobilized biologically active Get substance that has a prolonged effect. The Immobilization takes place until receipt of a biological active fermentation material with a fermentation content of 0.02 to 0.50 mass%.

The fabric thus obtained is squeezed out from the residue cleaned, dried, ge to the desired size brought and hermetically sealed, z. B. in hermetic sealed polyethylene packages packed with gamma Radiation sterilized in a conventional manner.

Before packaging in polyethylene film may be the material if necessary by known methods the required Be given shape, z. As cloths, Scharpie, powder, etc.

The inventive method is compared to the previously known with respect to the implementation of the Verfah rens and the equipment used here is simple and cheap.

By the method according to the invention, a material  be prepared that has the properties of a cure and a bandage. The material keeps furthermore all sanitary, physical and chemical characteristics of the original dressing material at.

The biologically active material obtained in this way is in packaged form when stored under normal conditions at least five years highly effective medically.

In the examples, the inventive method for various textile carrier described in more detail.

The material obtained in this way was under clini conditions in the treatment of 3200 patients tested on purulent necrotic disorders, Erfrie and trophic ulcers suffered.

The material of the invention was locally as follows applied: after the surgical treatment of the wound (Cutting out the purulent-necrotic mass, opening the Pockets, cutting through the bridges) and after formation a uniform cavity became physiological Solution soaked biologically active material in the form of Wipes or cotton wool (Scharpie) introduced.

It was found that in the treatment with the material according to the invention the wound after 15 ± 2.1 days healed while doing it with conventional treatment a control group of patients (3110 patients) with hypertonic solution and antiseptics lasted 26 ± 2.2 days. In other words, by applying the invention According to the material, the healing averages around 1.5fa accelerates.  

During the clinical application of the invention Materials were never toxic or allergic Reactions detected.

The material of the invention has because of the presence its one covalently linked to the surface Fer a higher degree of atraumatism. The power the effort to detach the material from the wound surface is half the size of a traditional non-modifi decorated dressing material.

The clinical trials of the material according to the invention showed high effectiveness in the treatment of Purulent or necrotic processes (a 1.3- to 1.8fa shortening the duration of treatment) to a high degree Atraumatic (the effort required to detach the material from the wound is half as large), a gerigeren ferment consumption (a 10- to 30-fold reduction in consumption native ferments in the treatment of the same disease a reduction of at least 25 times compared to known analogues (0.5 to 20% fermentation content in Analo ga against a 0.02 to 0.5% content in the invention proper material).

The verification of the stability of the invention Mate rials after sterilization by gamma rays shown that it is under storage in packaged form normal conditions (20 ° C) its healing effects as well all physical-mechanical and sanitary-hygienic Maintains properties for at least five years.

The material according to the invention has no side effects and contraindications.

Clinical efficacy using the erfindungsge  mäß biologically active material, eg. B. with immobilisier Hygrolytin, is compared to the traditional Methods of treatment of purulent-necrotic wounds of the Soft tissue is illustrated by the following table.  

table

The clinical treatment has shown that by the application formation of the textile material with immobilized ferments Patients with purulent surgical diseases Tissue the general treatment time by 1.3-1.8 times can be shortened.

Under the conditions of enzymotherapy, the Purification of the wounds and formation of granulations 1.7-2.1 times faster.

To illustrate the invention, concrete examples the method of preparation of the biologically active Stoffes and the bandage agent.

example 1

In a reactor 3.3 l of water are introduced. at 9.4 g of iodic acid are added to the running stirrer.

In another reactor, the same amount of water under Stir 1.6 g of sodium hydroxide. Both solutions are stirred for 5 to 15 minutes until the crystals of Acid and sodium hydroxide have completely dissolved.

Then both solutions are mixed and three to six Stirred for minutes. In this way you get a sodium periodate solution with a pH = 5.

In this solution, 1 kg woven fabric (med shear cotton gauze), which at room temperature 14 Hours stored in the dark (activated). After Activation, the substance (dialdehyde cellulose) is squeezed out, rinsed four times with 10 l of water and then out again presses.  

Activation activates in the cotton fabric functional groups, in this case aldehyde group at a concentration of 0.0625 mg eq. per gram of Textile support. To prepare the buffer solution, in this Case of the phosphate buffer solution will also be two reac used. In one, 3.3 liters of water with stirring with 5 g of potassium dihydrogen orthophosphate. In the second reactor are the same amount of water with stirring 37 g of sodium hydrogen orthophosphate added. It will be 10 to Mix for 20 minutes until the crystals are complete have solved.

Then both solutions are mixed in a reactor, whereby a phosphate buffer solution with a pH of 7.5 receives.

In the finished phosphate buffer solution is added 0.4 g of ferment, in this case hygrolytin, and the mixture is stirred 10 to 20 Minutes until the ferment has dissolved. In the received Solution is added 1 kg of the activated substance (Dialdehydcel lulose), and keeps it at room temperature for 12 hours. By covalent chemical bonds, the ferment is there immobilized. The fabric is rinsed until in the passed liquid no more protein detectable is. Then the fabric is squeezed out and 24 hours on the Air dried. In this way you get a material in this case, a cotton fabric that is biologically active and has a prolonged effect. The amount of immo fermented ferment, in this case hygrolytin, by the decrease in fermentation is determined in the solution is 0.2 mg per gram of cotton textile fabric, d. H. 0.02% mass%.

The dried material is ge in medical forms brought, for. B. in cloths required size. Then it will  packed in polyethylene double bags and in known Way sterilized with gamma rays of 25 kGy.

Example 2

Analogously to Example 1, prepare a sodium periodate solution ago. Add 80 g of medical cotton gauze and proceeds analogously to Example 1. This gives you a Material with an aldehyde group content of 0.78 mg eq. Per Grams of the fabric.

Thereafter, prepare a phosphate buffer solution as in example play 1 and mix it with 0.24 g of ferment, in this case Hygrolytin. The procedure is as in Example 1 with connect to form gauze bandages.

The amount of immobilized enzyme, hygrolytin, is 1.25 mg per gram of the textile carrier (0.125%).

Example 3

Prepare a sodium periodate solution as in Example 1, to which one gives 20 g medical cotton gauze and ver moves as in example 1. This gives you a material with an aldehyde group content of 3.125 mg-eq. per gram of the fabric.

Thereafter, prepared analogously to Example 1 a Phosphatpuf Ferlösung and mixed with 0.192 g of ferment, in this Case hygrolytin. It is continued as in Example 1.

The amount of immobilized enzyme, hygrolytin, is 5 mg per gram of the textile carrier (0.5%).  

Example 4

The procedure is analogous to Example 1, as fermented however Terrilytin used. The amount of immobilized Ferments, in this case Terrilytin, is 0.02% of the Mass of cotton gauze.

Example 5

The procedure is analogous to Example 1, as fermented however, collagenase is used. The amount of immobilized Ferments, in this case collagenase, is 0.02% of Mass of cotton gauze.

Example 6

The procedure is analogous to Example 2, as fermented however Terrilytin used. The amount of immobilized Terrilytins is 0.120% of the mass of cotton gauze.

Example 7

The procedure is analogous to Example 2, as fermented however, collagenase is used. The amount of immobilized Collagenase is 0.140% of the weight of the textile gauze.

Example 8

The procedure is analogous to Example 3, as fermented however Terrilytin used. The amount of immobilized Terrilytins is 0.5% of the mass of the gauze.  

Example 9

The procedure is analogous to Example 3, as fermented however, collagenase is used. The amount of immobilized Collagenase is 0.5% of the mass of the gauze.

Example 10

The procedure is analogous to Example 1, as a textile material however, cotton viscose gauze is used.

Example 11

The procedure is analogous to Example 2, as a textile material however, cotton viscose gauze is used.

Example 12

The procedure is analogous to Example 3, as a textile material however, cotton viscose gauze is used.

Example 13

The procedure is analogous to Example 1. For the production a phosphate buffer solution with a pH of 6.5 is added However, 20.7 g of potassium dihydrogen orthophosphate and 11.8 g Sodium hydrogen orthophosphate too.

Example 14

The procedure is analogous to Example 1. For the production a phosphate buffer solution with a pH of 7 is added 11.9 g of potassium dihydrogen orthophosphate and 23.6 g of sodium hy drug orthophosphate too.  

Example 15

The procedure is analogous to Example 1, as fermented however, trypsin is used.

Example 16

The procedure is analogous to Example 1, but is a Use phosphate citrate buffer solution with a pH of 7.5 det, which is manufactured as follows: In a first reactor is added 1.8 liters of water. When running 19.06 g of citric acid monohydrate are added to the stirrer.

In a second reactor with 8.25 l of water are added running stirrer 329.3 g of sodium hydrogen orthophosphate. you mixes until complete dissolution. Then both will Solutions mixed together.

Continue as in Example 1.

Example 17

The procedure is analogous to Example 13, but it will uses a phosphate citrate buffer solution, the following is produced in accordance with: In a reactor with 1.8 l of water is added while running Stirrer 71.7 g of citric acid monohydrate. In a second Reactor with 8.25 l of water is also given while running Stirrer 224.5 g of sodium hydrogen orthophosphate. After dissolution the salts, both solutions are mixed to give a Buffer solution with pH = 6.5. It becomes analogous to Bei continue game 13.  

Example 18

The procedure is analogous to Example 14. For the production a phosphate citrate buffer solution having a pH of 7 in a reactor with 1.8 l of water with stirring 38.1 g Citric acid monohydrate. In a second reactor with 8.25 l of water while stirring 290.5 g of sodium hydroxide genorthophosphat. after dissolution of the salts both Mixed solutions. Then, analogously to Example 14 continues method.

Example 19

A reactor is knitted with 50 l of 3M hydrochloric acid and 1 kg Tricotage fabric filled from polycaproamide fibers. The fabric 4 hours at a temperature of 60 ° C. To Termination of the hydrolysis, the hydrochloric acid is cooled and decanted. The knitted tricot fabric is in water rinsed until no more acid is detectable. Then fill the reactor with 40 l of 5% aqueous glutaraldehyde, adds 1 kg of hydrolyzed tissue, increases the temperature temperature to 50 ° C and let the material stand for 4 hours. Then the solution is cooled and decanted. The fabric is rinsed with water until the Glutaraldehydge smell is no longer perceptible. The activation of the Gewe bes is finished.

Through the activation form in the knitted tricot tissue 0.0625 mg eq. reactive functional groups, in this case aldehyde groups, per gram of textile fabric.

The phosphate buffer solution is prepared analogously to Example 1 provides. In this solution with a pH of 7.5 is added 0.66 g ferment, in this case terrilytin, and stir that Mixture 10 to 20 minutes until complete dissolution of the Ferment.  

Into the thus obtained terrilytin solution in the phosphate buffer (6.6 l) with a fermentation concentration of 0.01% is added 1 kg of the activated polycaproamide tricot fabric and leaves stand at room temperature for 9 hours. This is the ferment immobilized by covalent chemical bonds. Then The tissue is treated with physiological solution and with water rinsed, until the rinse water no ferment (protein) more is detectable. His amount can be known in per se Be controlled.

Then the tissue is squeezed out and left in the air for 24 hours dried. In this way you get a material, Nam a knitted biologically active polycaproamide Tricotage tissue with a prolonged effect.

The material contains 0.02% immobilized terrilytin (0.2 mg of ferment per gram of vehicle).

The dried material is transformed into medical forms, e.g. B. of cloths of the required size, brought. After that will it is packed in polyethylene double packets and in itself sterilized with gamma rays to 25 kGy.

Example 20

The activated polycaproamide tricotage fabric is analogous to Example 19 prepared; however, it will become 10% glutaraldehyde used hyd. The content of aldehyde groups in the tissue is 0.156 mg eq after activation. per gram of Tissue.

1 kg activated tissue is placed in a reactor with 6.6 l 0.03% Terrilytinlösung in phosphate buffer with a pH  of 7.5 and allowed to stand at room temperature for 8 hours. The Fabric is processed analogously to Example 19. The obtained biologically active material contains 0.6 mg Terri lytin per gram of tissue, d. H. 0.06% based on the Mass of the tissue. It is analogous to Example 19 weiterver drive.

Example 21

Prepared analogously to Example 19 activated polycapro amide tricot fabric but uses 15% glutaraldehyde hyd. The content of aldehyde groups in the tissue is after the activation 0.26 mg eq. per gram of tissue.

1 kg of activated tissue is treated with 6.6 l of 1% Terrilytinlö solution in the phosphate buffer with a pH of 7.5 and poured over allowed to stand at room temperature for 10 hours. It will proceed analogously to Example 19.

The resulting material contains 2 mg terrilytin per gram the wearer, d. H. 0.2% based on the mass of the tissue Ferment.

Example 22

The procedure is analogous to Example 19, but as Fer used trypsin. The resulting material contains 0.02% immobilized trypsin on the textile support, in in this case on knitted polycaproamide tricot fabric.

Example 23

The procedure is analogous to Example 20, as fermented however, trypsin is used. 1 kg of activated polycaproamide Tricotage tissue is supplemented with 6.6 L of 0.03% trypsin solution  Poured phosphate buffer and at room temperature for 9 hours ditched. The procedure is analogous to Example 20.

The resulting material contains 0.06% trypsin based on the mass of the knitted polycaproamide tricot fabric.

Example 24

The procedure is analogous to Example 21. The immobilization However, this is done in 0.15% trypsin solution in phosphate buffer.

The material obtained contains 0.3% trypsin based on the Mass of the tissue.

Example 25

An acid hydrolyzed polycaproamide tricot tissue is behaved in a reactor in dimethyl sulfate in the liquor of 10 at room temperature allowed to stand for 10 hours. The excess dimethyl sulfate is poured off. The fabric is once with ethyl alcohol in the liquor ratio of 20 rinsed at 4 ° C, then three times with the phosphate buffer of pH = 7.5 at 4 ° C. The fabric thus obtained has reaction capable imido ether groups.

Immobilization of the enzyme, in this case trypsin ins, takes place in a liquor ratio of 10 from one Ferment solution with phosphate buffer at one concentration of 0.04%. The procedure is carried out at room temperature leads and lasts 16 hours.

Thereafter, the substance with physiological Natriumchloridlö and rinsed with water until no more fermentation is detectable in the water. His amount is in itself controlled manner known.  

After that, the fabric is squeezed out and left in the air for 24 hours dried. This will be biologically active material, in knitted polycaproamide tricot fabric in this case, receive a prolonged medical effect has. The material contains 0.2% immobilized trypsin, d. H. 2 mg per gram of vehicle.

Example 26

The procedure is analogous to Example 25, as fermented however, trypsin in a 0.008% trypsin solution in Phos phat buffer in the liquor ratio of 10 twelve hours immo stabilizes. This is a material with a 0.04% Content of immobilized trypsin, d. H. 0.4 mg per gram of the wearer.

Example 27

The procedure is analogous to Example 25, but the Trypsin in a 0.024% trypsin solution in the phosphate buffer immobilized in a liquor ratio of 10 ten hours. The Material contains 0.12% trypsin, d. H. 1.2 mg trypsin per Grams of vehicle.

Example 28

The procedure is analogous to Example 19, as fermented however, lysozyme is used. The material contains 0.02% lyso zyme.  

Example 29

The procedure is analogous to Example 20, as fermented however, lysozyme is used. The material contains 0.06% lyso zyme.

Example 30

The procedure is analogous to Example 21, as fermented however, lysozyme is used. The material contains 0.2% lyso zyme.

Example 31

The procedure is analogous to Example 19, as fermented However, protosubtilin used. The material contains 0.02% Protosubtilin.

Example 32

The procedure is analogous to Example 20, as fermented However, protosubtilin used. The material contains 0.06% Protosubtilin.

Example 33

The procedure is analogous to Example 21, as fermented However, protosubtilin used. The material contains 0.2% Protosubtilin.

Example 34

The biologically active ester obtained analogously to Example 1 Rile material is a woven medical cotton gauze  with immobilized and covalently linked thereto Hygrolytin. The material contains 0.2 mg hygrolytin per Grams of woven cotton gauze.

The material in the form of medical towels can for Treatment of abscesses, phlegmon and postoperative Complications of surgical sutures are used.

When using this biologically active material is a prolonged medicinal effect of the enzyme up to Scored 48 and even 96 hours while the native Fer ment used to treat the same condition will work, at most one hour. When treating with This biologically active material is the complete Healing the wound in 17 days. In individual cases feel The sick in the first hours after the creation of the Association a slight burning and irritation of the wound. all There were no allergic reactions.

The covalent nature of the bond between the carrier and the ferment of the biologically active material used is based on the maintenance of fermentative Aktivi after treatment of the material with saline solutions, z. B. with sodium chloride, or with buffer solutions, eg. B. with Phosphate buffer, detected. The physically adsorbed Proteins (ferments) are in this case from the carrier surface washed out.

The material retains its medicinal properties normal storage conditions for 5 years.

Example 35

The biologically active mate obtained analogously to Example 1 Rial has a composition as in Example 1, contains  but 1.25 mg hygrolytin per gram of woven cotton Mull. In the medical application of this gauze with the immobilized hygrolytin, which is covalently linked to it is the duration of the healing effect 48 to 96 hours. By contrast, the native ferment is only effective for 1 hour.

When treating with this biologically active material Complete healing of the wound occurs in 15 days. The other results are analogous to Example 34.

Example 36

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains 0.5 mg hygrolytin per gram of woven cotton gauze. at the treatment with this biologically active material occurs complete healing of the wound in 14 days. The other results are analogous to Example 34.

Example 37

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains as a textile carrier knitted medical cotton gauze. The treatment results are analogous to Example 34.

Example 38

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains as a textile carrier, non-woven cotton fibers. The treat results are analogous to Example 34.  

Example 39

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains Linen woven as a textile carrier. The treatment results are analogous to Example 34.

Example 40

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains as a textile carrier natural silk. The treatment results are analogously to Example 34.

Example 41

The biologically active material obtained as in Example 19 has a composition as in Example 19, but contains as a textile carrier knitted polycaproamide fabric. The Be action results are analogous to Example 34.

Example 42

The biologically active material obtained as in Example 19 has a composition as in Example 19, but contains as a textile carrier knitted polyurethane fabric. The treat results are analogous to Example 34.

Example 43

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains as a textile carrier knitted viscose fabric. The treat results are analogous to Example 34.  

Example 44

The biologically active material obtained as in Example 1 has a composition as in Example 1, but contains as a textile carrier knitted sec-acetate cellulose fabric. The treatment results are analogous to Example 34.

Example 45

The biologically active material obtained as in Example 21 has a composition as in Example 21, but contains 2 mg lysozyme per gram of the textile carrier. The material with the immobilized lysozyme that is covalent with the carrier has a prolonged effect of 48 to 96 hours compared to a one-hour duration of action with native ferment on. When treating with this biologically active material occurs a complete cure the wound after 15 days.

The treatment of purulent wounds in the phase of hydration with the immobilized lysozyme-containing textile material contributes to the faster elimination of wound infection faster dosage of alternative purulent inflammation, for purifying the wounds of purulent necrotic tissues, for stimulating the synthesis of glycogen and nucleic acid ren (DNA and RNA) in the cells of the wound, to the active Proliferation of connective tissue cells (fibroblasts), to Reinforcement of collagenosis and faster Verklei irrigation and contraction and epithelialisation at the wound.  

Example 46

The biologically active material obtained as in Example 1 has a composition as in Example 5, but contains 0.2 mg collagenase per gram of the textile carrier.

When treating with this material, the complete the healing of the wound after 16 days. The remaining Treatment results are analogous to Examples 34 and 45th

Example 47

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The material is shaped as a shard with 1.0 mm fibers and is suitable for the treatment of deep purulent puncture wounds and inflammation in the rectum.

When treating with this material, a complete healing of the wound after 17 days.

Example 48

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The material is shaped as a sharpie with 8.0 mm fibers and is suitable for accelerated treatment of infection herds in teeth (chronic granulating periodontitis).

The Scharpie formed material has an anti-exudative and promotes the effluence of the exudate via the root canal, which even tortuous and narrow, but definitely must be permeable.  

Example 49

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The material produced as a share with 15 mm fibers is suitable for the treatment of inflammatory processes in the Rectum.

The healing effect occurs after 16.5 days.

Example 50

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The powdered material having a particle size 0.001 mm is suitable for postoperative treatment Seam complications.

When using this powder, the biological, including Fermentative, active, complete healing occurs Suture after 17 days.

Example 51

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The powdered material having a particle size 0.5 mm becomes purulent-necrotic for treatment Processes with complicated localization as well as difficult accessible cavities used.

The healing effect occurs after 17 days.  

Example 52

The biologically active material obtained as in Example 15 has a composition as in Example 1.

The powdered material having a particle size 1.0 mm in diameter is used to treat phlegmons.

The healing effect occurs after 16.5 days.

Example 53

A composed of a three-layer bandage and an element for Sticking (tape) existing bandage means sets consisting of a protective layer, an absorption layer and a wound contact layer together. The latter will be out the biologically active mate obtained as in Example 21 rial produced.

This dressing was used in the first care at a Injured soft tissue with a size of 6 × 4 cm and a depth of 4 cm. At the delivery The patient was admitted to the clinic 12 hours after Trauma the absence of edema or of Perifokalentzün firm. The inner surface of the wound was clean, that Tissue viable. A complete healing of the wound entered 13 days after surgical treatment.

Example 54

A composed of a three-layer bandage and an element for Sticking (tape) existing bandage means sets  consisting of a protective layer, an absorption layer and a wound contact layer together. The wound contact layer is obtained from a biological as obtained in Example 21 active material, the absorption layer of a like in Example 30 obtained biologically active substance Herge provides.

The bandage was used in the initial treatment of an injury of the soft tissues of the leg 4 cm long, 3 cm wide and 2 cm deep. When the patient was admitted to the clinic, no edema or perifocal inflammation was noted 8 hours after the injury, the inner wound surface was clean, and the tissue was viable. After surgical dressing with the dressing, complete healing of the wound occurred after 11 days.

Example 55

A bandage made from a three-layered bandage a plaster is as a fastener was manufactured. The bandage has one adjacent to each other Protective layer, an absorbent layer and a wound con on shift. The wound contact layer consists of a analogously to Example 21 produced biologically active Fabric, the absorption layer from an analogous to Example 30 produced biologically active substance in the form from Scharpie with a fiber length of 10 mm.

The bandage was used for primary wound care in case of soft tissue injury (size of the injury 5 x 2 x 3 cm).

When the patient was admitted to the clinic, 10 No edema or inflammation hours after the injury  found. After surgical wound care with the dressing occurs a complete healing of the wound after 11.5 days.

Claims (19)

A biologically active material containing a vehicle and an immobilized thereon ferment, characterized in that it comprises a fabric as a carrier and this fabric contains a covalently associated with him ferment, wherein the ratio between the ferment and the carrier indicated as follows becomes: ferment 0.02 to 0.50 mass% carrier 99.98 to 99.50% by mass 2. Biologically active material according to claim 1, since characterized in that the textile material consists of natural fibers. 3. Biologically active material according to claim 1, since characterized in that the textile material consists of synthetic fibers. 4. Biologically active material according to claim 1, since characterized in that the textile material consists of synthetic fibers. 5. Biologically active material according to one of claims 1 to 4, characterized in that the fabric is a woven or non-woven or a knitted fabric is. 6. Biologically active material according to one of claims 1 to 5, characterized in that it contains as ferments proteolytic ferments. 7. Biologically active material according to one of claims 1  to 5, characterized in that it contains collagenolytic ferments as ferments. 8. Biologically active material according to one of claims 1 to 5, characterized in that it contains bacteriolytic ferments as ferments. 9. Biologically active material according to one of claims 1 to 8, characterized in that it in the form of Scharpie with a fiber size of 1.0 to 15 mm is present. 10. Biologically active material according to one of claims 1 to 8, characterized in that in the form of a powder with a particle size of 0.001 to 1.0 mm is present. 11. Process for the preparation of the biologically active mate Rials according to one of claims 1 to 10, the Vorberei tion (activation) of the carrier, the immobilization of the Ferments on the carrier using a buffer solution solution, the subsequent rinsing and drying and the position of medical forms from the material obtained provides, characterized that the chemically inactive carrier, in this case a Tex tilträger, to form reactive functional Groups containing 0.0625 to 3.125 mg eq. Per Gram of the carrier is activated, the biologically active Substance, in this case a ferment, from a buffer solution with a pH of 6.5 to 7.5 at about room temperature in 8 up to 16 hours on the preactivated carrier under formation covalent chemical bonds between the textile carrier and the enzyme is immobilized, the material obtained pressed in a conventional manner and ge with water is rinsed until the rinse water is fermentation free, the materi  al is dried at about room temperature (about 20 ° C), hermetically packaged and sterilized. 12. The method according to claim 11, characterized ge indicates that the sterilization with the help of gamma-rays in doses of 25 kGy. 13. The method according to claim 11 or 12, characterized characterized in that a cellulosic Textile carrier by oxidation with alkali periodate to Formation of dialdehyde cellulose is activated, which is 0.0625 to 3.125 mg eq. reactive functional groups contains per gram of the carrier. 14. The method according to claim 11 or 12, characterized characterized in that the cellulose is not containing textile carrier made of plastic acid acid hydrolytic with subsequent rinsing in water and by the reaction of the hydrolyzed carrier with a solution a reactive functional group-containing Stoffes to achieve a content of reactive Groups from 0.0625 to 0.3125 mg eq. per gram of the vehicle is activated. 15. The method according to claims 11 or 12 and 14, since characterized in that the hydroly Said carrier mixed with a glutaraldehyde solution and then rinse with water. 16. The method according to claims 1l or 12 and 14, since characterized in that the hydroly Sized carrier is mixed with a dimethyl sulfate solution and then rinsed with ethyl alcohol and with phosphate buffer becomes.   17. Dressing material consisting of a three-layer dressing with a protective layer, an absorption layer and a Wound contact layer is characterized ge indicates that there is at least one layer from the biologically active material according to one of the claims che 1 to 8. 18. dressing according to claim 17, characterized ge indicates that the wound contact layer The biologically active material according to any one of claims 1 to 8 and the absorbent layer of a textile material, that does not contain a biologically active substance is. 19. dressing according to claim 17, characterized ge indicates that the wound contact layer The biologically active material according to any one of claims 1 to 8 and the absorption layer of the biologically active Material according to one of claims 1 to 5 and 7 to 10 is made.