CA2011220A1 - Material having biological activity method for preparation thereof and dressing - Google Patents

Abstract

BIOLOGICALLY ACTIVE MATERIAL, METHOD FOR PREPARING SAME, AND DRESSING MEANS A biologically active material comprises, as a carrier, a textile maternal, and an enzyme, immobilized on said tarrier and covalently bound thereto, said components being present in the following weight ratios, per cent by weight: enzyme - 0.02 to 0.50, carrier - 99.98 to 99.50. A method for preparing said material resides in activating said textile material until the formation therein of reactive functional groups in an amount of from 0.0625 to 3.125 mg-equiv. per gram of the carrier, immobilizing on said carrier an enzyme taken from a buffer solution having a pH of from 6.5 to 7.5 at room temperature for 8 to 16 hours to form a covalent chemical bond between said textile material and enzyme, squeezing-out the material thusobtained, followed by washing the material, drying it, shaping it into medicinal forms, placing it into a container, sealing it off, and sterilizing An individual dressing means is made in the form of a bandage consisting of , arranged in succession therein, a protective layer, an absorbing layer, and a woundadjoining layer. At least one of said layers is made of said biologically active material.

Description

2~1122~

BIOLOGICALLY ~CTIVE ~TERIAL, ~I~THOD FOR ~RE;PARI~G
SAME, AND DRESSING M~ANS

The present invention relates to the art of medicine and, more specifically, to a biologically active material, to a methc , `
`~ for preparing said material, and to a dressing means based on said material.
i The present biologically active material may be widely used for treating traumata, frostbites, trophic ulcers, pyo-necrotic wounds,and diseases which are accompanied by the formation of necrotic sections and by accumulation of thiek, viscous exudate, and it may be applied in surgery, dentistry, urology, proctolog~
. ~astroenterology. Furthermore, the present biolo~ically active material is applicable in veterinary, biotechnology, for instan-; ce, as a sorbent for recovering trypsin~ antitrypsin inhibitc ;, State of the Art It. i8 known in medical practice to use native proteolytic en7.yme preparations for treating pyo-inflammatory diseases.
However, æuc.h enzymes are expen~ive and in short supply. Regard-less of their certain effectiveness, such enzymes are not . . .
resi~tant against inhibitors which are rontained in wound di~-charge, nor are they resistant against pH and temperature varia-tions. Moreover, said enzyme~ posses~ antigenic (aller~ic) and pyrogenic properties.
~ There is known in the prior art a film material (Cf. SU

Inventor's Certificate No. 700,138, issued on November 30, 1979 having biological activity and containing a carrier, such as -.,~' ' ' ':

-2- 201~22~

cellulose triacetate or secondary acetate, and a proteolytic enzyme, such as, for instance, tryp~in, ~ -chymotrypsin, said components being taken in the following ratio:
cellulose triacetate (secondary acetate) - 80 - 99a/1 by weig~
enzyme - balanc-s.
The above-cited film material is prepared by emulsifying ~-an aqueous solution of a proteolytic enzyme in a cellulose ester solution in a water-immiscible organic solvent, followed by forming a film. In the film material thu~-prepared, the proteolytic enzyme occurs in the form of finest inclusions.
The above-described material is capable of producing a long- -sustained necrolitic e~.fect due to gradually releasing small doses of the enzyme from the film surface. '~'7''_'._..
However, once the enzyme has been releas~d from the materia]
the ehzyme exhibits all the properties of a native material, including all of it~ disadvantages, namely, non-stability against inhibitors, against pH and temperature variations, antigenecity and pyrogenecity. And finally, it is necessary to note that the enzyme consumption rate is high - up to 20 by weight.
Application of this material is sestricted by the high pro-bability rate for enzyme inactivation in organic solvents which are used for forming films or monofilaments. The mate-rial itself can be used only as a draining material~ and it cannot be used as a dressing material.
Also known in the art ~s a granular material (Cf., e.g.
laid-open French Patent No. 2,556,222; issued on June 14,1986) :.:
:`~t .A: , . ' : , ~3~ 201~22~

possessing a biolo~ical activity and containing a carrier, such as dextran, chitin, chitosan, and, immobilized thereon9 an en7.yme, such as chymotrypsin, trypsin, said components being present in the following weight ratios: carrier - 80.0~ 80.5 weight, enzyme - balance. The abo~e-cited material exhibits a prolonged action, and the enzyme contained in it has no possi bility to penetrate into a blood channel and manifest antige-necity.
The cited material is also disadvantageous in that it has an elevat~ed enzyme content, namely, up to 20~ by weight, and i9 expensive. Apart from it, this material calls for the use of special installations and apparatuses for its storing in specially prepared buffer solutions at lower temperatures (at 2C) -Equally known in the art is a method for preparing a biologi cally active material (Cf. SU Inventor~ Certificate-No. 700,_ ;~ 138; issued on November 30, 1979), which comprises the follow-ing steps of:
- preparing an aqueous solution of a proteolyti~ enzyme, preferably, trypsin;
- preparing a solution of cellulose ether~ in the present ; case, cellulose triacetate, in a water-immiscible organic solvent;
- emul~ifying the former solution in the latter one;
:::, - :.
i - shaping the material into medicinal forms, such as plates, J
films, etc., using a dry-forming method;

- holding the medicinal forms thus-obtained for 1 hour in a .
. :

2011~2~

; vacuum drier at a temperature of 25 C.
The materials prepared ~y the above-cited method possess a 1~iological activity and contain components in the ~ol]owing weight ratio~, in per cent by weight:
- cellulose triacetate or ~econdary acetate - 80 to 99 - trypsin - balance (1 to 20).
The latter method suffers from a number of serious disadvan-tages:
- a considerable enzyme consumption rate (up to 20%3;
- a partial loss of the enzymatic activity during emulsifi-cation in an organic solvent;
- absence of a chemicaI bond between an enzyme and a carrier owing to the fact that the enzyme is physically incorporated into the structure of the material;
- release of the enzyme into wound discharge in the form of a native preparation, which may result in a negative side effect, such as, for instance, allergic reactions; and - need for the use of specially designed equipment, such as a vacuum drier.
The material prepared by the~ above-cited method in the form of plates cannot be used as a dressing material, since it does not possess the requisite sanitary and hygienic properties, such as hygroscopicity, air permeability, elasticity, etc., and it can be used only as a draining material. -Also known in the prior art is a method for preparing a - biologically actiYe material (Cf. laid-open French Patent ~o.
2,550,222, issued on June 14, 1985; laid-open F.R.G. Patent , . . .
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201~22~

No. 3,444,746, issued on June 20, 1935; and Czechoslovakian Inventorfs Certificate No. 249,311, issued in 1988), which com-prises the following steps of:
- preparin~ particulate material (carrier), such as cross-linked dextran, chitin, chitosan, by causin~ them to swell in water;
- preparing.an ac.tivating solution of 2-amino-4,6-dichloro--s-triazine in acetone, followed by dilution with water at 50 C
- activating the material with an activating solution at 50 C under stirring conditions;
~` - adding to the activating solution containing said material . an aqueous solution of sodium carbonate and hydrochloric acid and stirring at 50 C;
- filtering the resmlting mixture;
- washing the remaining material with an aqueous solution of acetone and water to obtain an ac.tivated material which can be stored in a phosphate buffer at 2 C;

., ~. .
- preparing an enzyme solution, such as, for instance, a ' chymotrypsin solution~ in a bu~fer; ~

.~ - immobilizing the enzyme on the activated material~ such ~

cross-linked dextran, chitin,.chitosan; ~.
:::
- washing the material containing, immobilized thereon, ~:
chymotrypsin. in turn, with borate and acetate buffer solutionc containing sodium chloride at different concentrations; ~ .
- washing the material with a borate.buffer solution without .
sodium.chloride; and -lyophilizing the material with the enzyme immobilized there~
~, :

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The above-cited methods made it pos3iblc to produce biologi-cally acti~e materials~ their components being taken in the following weight ratios~ per cent by weight:
. - enzyme - 19.5 t.o 20 - carrier - balance.
The above method suf~ers from the following essential limi-tations:
- a considerable enzyme consumption rate (up to 20%);
- a multi-step production technology and, as a result, poor efficiency and high cost;
. - need for the use o~ short-supply and e~otic raw materials, .. . ,.~ -~
. such as, for instance, chitin obtainable from crab shells or ....
. from myce~lium of Penicillium chrvsogenu~ fungus; and ~.i ~; - need. for the use of specially designed equipment for ': ~toring the acti~ated material and for carrying out the. lyophi- .

.:~ lization step.

" The biologically active material prepared by the above-cited ~ method.cannot be used as a dressing one, and i.t is usable :~ for~ of a ~I only in th~ powder or a suspension,. since in its finiished form .. ~ the cited material represents a powder containing an immobilize .~:
, I -enzyme having a particle size.o~ from 0.01 to 0.5 mm. .

There is known in the prior art a biologically active dress~

: ing means (C~. British Patent No. 2,092,006, ~sssed on Februar~

.`1 11, 1986~, comprising, arranged in succession therein, a pro-. . - . .
tective layer, an absorbing layer, and a wound-adJoining layer.

-, This dressing means comprise~ a thin layer of the enzyme applied to the internal side o~ the wound-adjoining layer. The ,,~
' " ~ -:
~ .

, . ~: .: : : : . .. . - . , . .:

~7~ 201~22~3 protective layer can be made in the form of a woven or non-~over material.
The absorbing layer can be made from cotton, or viscose (cot-ton wool)~ or rayon.
The wound-adjoining layer i.s made in the form of woven or non-woven textile materials~ or perforated polyester, polyamide, polyurethane, polyethylene, polypropylene and cellulose triace-tate film, and it contains metallic copper or copper compounds.
~' As the wound discharge comes to contact with the above-cited dressing means~ all of the negative properties of the I enzyme become manifest as those of a native enzyme, in particu-lar, :~t~s~ instability against inhibitors which are contained in the wound discharge, against pH and temperature variations~
' antigenecity aIld pyrogenecity, high cost owing to a high enzyme 3i, consumption rate.
Brief Desciption of the.Essence of the Invention .; It is the object of the present i~lvention to develop smch a :
biologically ac.tive material as would ensure the prolonged J
- action of the medicinal substances contained therein, be amenabl ~, to storing for a long time in packages in air-dry state under normal condition~, while retaining all the sanitary and hygienic propertie~ characteristic of a dressing material, would secure a medical effect within a shorter period of treatment at a ~ lower content of a biologi¢ally active principle, and would be ` capable of being effectively used as a dressing means.
Another object of the present invention is to develop such : ~
a method for preparing a biologically active material as would .

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enable the production of a l~laterial ~or use as a dreissing means : wo~lld be technologically simple, would not be expensive and would not call for high material ~xpenses due to use of readily available materials, and could be easily adaptable under commer cial-scale production conditions.
A further object of the inventio.n is to provide a dressing . mean ~ hich would ensure a high medical effect at a considerably i smaller enzyme consumption rate, would be resistant to inhibi-tors which are contained in the wound discharge~ to pH and temperature variations, and would be free from antigenic and pyro~enic properties.
~ The abo~e-formulated objects ha~e.been achieved by providing -.~ a biologically active material compristng a carrier and, immo-`I bilized thereon, an enzyme., which material, in accordance with ~-.~ , , the invention,. comprises, as said carrier, a textile material, .~:
., :
while said enzyme is: bound to said carrier by a co~alent bond~

.: the componentsi of the material being present in the following ~ weight ratios, per cent by weight~
.
.~ - enzyme - 0.02 to 0.50 :::
i~ ,: ::
q - carrier - gg.98 to 99.50 .~ The textile material is used fo~ the reason that it has a ~
~- weakly pronounced: medical effeet, since it ïs capable of drain- -. ing the wound~ of adsorbing microbes and:toxins~ of prote¢ting the wound surface from contamination, and of preventing from ~ ~
.:i exce3sive loss of moisture. ~ ~.
The above-specified weight ratios are determined by the fact that the use of a biologicQlly active material containing :~

, I
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: j . .......... - -~,. : :. . ~ : ~ , -: :: .

9- 20~122~

an enzyme in an amount of below 0. 02~o by weight ~harply reduces the medical effect (by more than a factor of 1.5), whereas the use of a material containing more than 0.50$ by weight of an enzyme does not considerably shorten the duration of treat-ment (an average treatment period lasting 14 + 2 days), but, instead, sharply augments the cout of the material and treatment.
In medical practice, it is advisable that the period of a single application of the claimed material to the wound be from ., .
48 to 96 hours. - -The medical efficacy of the present material, if it is store in packaged state under normal conditions (i.e. at room tempera ture of about 20 C), is retained for a period of not less than 5 years.
The use of the material in accordance with the~present -invention makes it possible to shorten the period of healing wounds by an average factor of 1.5 at doses which are 10 to 30 times smaller-than those~ normally u~ed for healing wounds by means of the conventional biologically active materials or native enzymes. The claimed material is not toxic, nor does it cause any allergic reactions.
It- is ad~isable that the textile material would represent a woven, or non-woven, or knitted fabric.
It is possible that the textile material be made of natural .:... . . .
fibres-, which provides for better conditions for application , of the present material onto the wound surface, close to com-.' :
~ fortable conditions.
.

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_ 1 ir)_ 201122~

In order to e~pand the range of raw materials potentially amenable for the use by the present inveintion, it is adivsable ' that the textile carrier be made of ~ynthetic or man-made ~i~re i It is also advisa~le that the material in accordance with the present invention would contain, as an enzyme, proteolytic, or c.ollagenolytic~ or bacteriolytic en~ymes, since these enzyme make it possible to split denaturated proteins having various , structures.
l~ It is also advisable that the present biologically active ~.
material be turned out in the form: o.f lint: having fibres 1.0 ~l~ to 15 mm long~ whereby it becomes possible to use the material .
; in stomatology and for treating deep wounds, including puncturec :~
i purulent wounds o~ Gomplicated configurations.
' It is also possible that the p~esent material be turned ::
out in the form of a powder ha~ing a particle size of from 0.001 to 1.00 mm, whereby it becomes possible to use it for .i treating extensive wound sur~aces of complicated configurations ~'i The present material is suitable. for treatment of pyo-ne~ro~
`~ tic wounds, acute and chronic forns of periodontitis, wounds having complicated configurations and cavities, hollow organs ~ of a live organism having narrow excretory channels, for ~: instanc.e, for treating inf?ammatory processes in the urinary bladder cavity.
The approbation of the.present ~aterial under the conditions of tests and clinical observations ha~ demonstrated its high biological and therapeutic activityl Special studies have shown .
, that the present material shows a peak of its therapeutic .. , :

... .

~ - 20~22~
.~

activity, when applied onto the wound surface, for 4 days from the moment of application.
The test animals have shown no signs of allergic reactions after the application o~ the biologically active l~-aterial : . :
prepared in accordance with the present invention. Nor have been observed such changes in the hemostasis functions as the level o~ fibrinogen degradation products~ fibrino~en content, aad fibrinolytic activity.
The above-described biologically active material is prepared by a method comprising the steps of activating a carrier, immo-bilizing thereon an enzyme using a buffer solution, followed by washing, drying and shaping the material thus-prepared into medicinal forms. In accordance with the present invention~
the step of activating a carrier, namely, a textile material, is carried out by forming therein reactive (reaction-capable) functional groups until the~amount of such groups reache~
0.0625 to 3.125 mg-equiv. per gram of the carrier. The immobi-lization treatment of the enzyme is carried out from a buffer solution having a-pH of from 6.g to 7.5 at room temperature for 8 to 16 hours to form a ~ovalent chemical bond between the textile material and enzyme, whereupon the material thus-prepared is squeezed out, washed until no enzyme traces are detected in the washings, dried at room temperature (at about 20 C), placed into a container, and sterilized.
The present method is distinguishable over the prior-art ones in that it is simple and cheap in its realization in term of its engineering aspect and the equipment required for its implementation.

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201~ 22~
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~ The claimed method ~lakes it possible to prepare a material which conllines the properties of a rnedicinal pr0paration with those of a dressillg means, and which retains all the sanitary--and-hygienic and physico-chemical properties of the starting dressing ;naterial. :~
~ The biologically acti~e material thus-prepared retains its ~ ~
.~ hi~h therapeutic efficacy while being stored under normal ~ p conditions in packaged state for a period of at least 5 ye 9, It is advisable that acti~ation of a c.ellulose-containing : :
.
~ , ~' textile carrier be carried out by subjecting it to oxidation :~
. ~
` with an alkali metal periodate to ~orm cellulose dialdehyde containing reactive functional aldehyde groups in an amount .
of from o.o6z5 to. 3.125 mg-equiv. per gram of a carrier, ~ It is preferable that activation of a textile carrier made `' of synthetic fibres be conducted by subjecting the carrier to hydrolysis with acid, followed by washing the carrier thus-~ hydrolyzed and by reacting it with.a solution containing reacti~
`'! funct.ional groups until the amount of the latter in the carrier ~ reacles from o,o625 t.o 0,.3125 mg-equiv. per gram of the carrier :l It is advisable that the treatment of the hydrolyzed carrier made of synthetic. fibres be carried out either with.a glutaric .~ aldehyde ~olution, followed by washing with water, or with a :;~
dimethyl sulphate solution, followed by washing the carrier :~
.~ in ethanol and in a phosphate buffer solution. ~. -. ., The above-formulated objects are accomplislled owing to ~he fact that in a dressing means in the form of a bandage compris-ing, arranged in succession therein, a protecti~e layer, an ~-~' `, r _13_ 20~122~

absorbit~g layer, and a ~ound-acljoining layer, in accordance with the invention, at least one l~yer is made from the above-, described biolo~ically active material.
! It i9 advisable that in said dressing means the wound-adjoin-A . , ing layer be formed from the above-described biologically active material, while said absorbing layer be formed from a textile material.
It is preferable that in the dressing means the wound-adjoin-ing layer be formed from said biologically active material, ~-in which proteolytiOE, or collagenolytic~ or bacteriolytic enzymes are used, while the absorbing layer be also formed from said material, in which collagenolytic or bacteriolytic enzymes are used, the enzymes used in both of said layers being different from each other.
Application of the above-described dressing means during the period of a few hours after a trauma (an injury~ makes it possible to prevent it from infecting and to shorten the period of subsequent treatment by 2 to 4 days.

Detailed Description of the Invention , The biologically actiYe material comprises, as a carrier, a textile material and an immobilized enzyme which is bound to this textile material by a covalent bond, the enzyme and textil~
material being present in the following weight ratios, per cent j by weight:
- enzyme - 0.02 to 0.50 - textile material - 99.98 to g9.50.

The textile material may be made from natural fibres, such a 2~22~

for instance, cotton, linen, silk, wool fihres, or from synthe-tic fibreg, such as, for instance, polyacrylonitrile, polycapro : ~:
amide, polyurethane fibres, or from man-~ade fibres, such as, ~:
for instance~ triacetate, celllllose secondary acetate, viscose ~ fibres.
The textile material may be in the form of, for instance, :. woven, or non-woven, or knitted fabric.
As said enzymes~ the present material may make use~ of proteo~-. lytic enzymes, such as, for instance, hygrolytin, protosubtilin terrilytin, trypsin, or collagenolytic enzymes~, such as, for ~; instance, c.ollagenase, or bacteriolytic enzymes, such as, for instance, lysozyme.
The specific choice for practical application of the abo~e-mentioned groups of immobilized enzymes is conditioned by their therapeutic properties.
It. is advisable that the proteolytic enzymes, such as, for instance, trypsin, be applied at the hydration stage of treat-ment of pyo-necrotic woundY, that is, when it. is necessary to.
, split (cleav~) necrotic masse~ to liquefy viscous dense exu-;i date, etc.
~ The bacteriolytic enzymes, such as, for instance, lysozyme~
.. are recommended; for suppression of microbial growth in the woun The collagenolytic enzymes, suc.h as, for instance, col].agena se, exhibit both proteolytic and collagenolytic activity and~
as such~ they are particularly effective in treatment of burn-~1 induced diseases.

In the light of the foregoing, it will be understood that :~

~r.
,1 ~d .~j j.:: . ' ~ -15-20~22~

in view of the extrei11e variety of various pyo-necrotic and inflammatory processes, there i9 a need for biolozically active materials containing immobilized enzymes having different natur and therapeutic properties.
Dependin~ on the con~iguration of a wound or a channel through which the biologically active material i3 supplied to the wound surface~ said material may assume various ~edicinal forms, namely, cloth, banda~es, napkins, lint having fibres 1.0 to 15 mm long, and powder having particles with sizes rang-ing from 0.001 to 1.0 mm, The study Q~ bacterial inoculations from wounds prior to and after the use of the present material made it possible to arrive at well substantiated conclusions, to-wit: the range of inoculated microorganisms bec~me narrower, the virulence of the sown microorganisms weakened due to their substitution by a less pathogenic flora, their resistance to antibiotics weakened. Application of the present material lowers the level ..
of bacterial dissemination of wound surfaces, the period of preparing patients for plastic operations and the period of staying of patients at stationary hospitals become shorter by 10 days on the average.
During the entire period of treatment, patients~' urine and blood were analyzed, their temperature was taken. None of the cases showed any sign~ of general or local complications which would be caused by the application of the present material.
The present material underwent testing under clinical condi-tions for treating 3200 patients affected with pyo-necrotic .:
~ -16-~ 20~ 122~

diseases, frostbites, trophic ulcers, etc.
The present material was ap~plied locally following the pro-cedure as described below: after a preliminary surgical treat-ment, such as dissection of the pyo-necrotic area, opening of .
pockets, dissection of intersections and for~ation of a single cavity, the proposed biologically active .naterial, for instance, . in the form.of napkins, cotton wool (lint) impregnated.with a physiologic salt solution, was incorporated into the wound.
It has been established that, when the present material was ~. applied, the period of healing the wounds was reduced to 15 + 2.
days, whereas during treatment by a eonventional method using a hypertonic salt solution c.ontaining antiseptics of a control ~:
batch of patients (3110 patients~, this period was 26 + 2.2 days~ that is, application of the present material makes it possible t:o accelerate the c.ourse of treatment by an average ., ~j factor of 1.5.

.~ None of the cases of clinical application of the present j",~l .
j material showed. any toxic: or allergic reactions.
The present material i9 highly atraumati.c due to the~presenc( on its surface of a co~alently bound enzyme. The effort require : 'l :~f for breaking off the present material fro~l the wound surface ~.~ is on the average half as that of the non-modified commonly :. applied dressing material.

j The results of the clinical application of the claimed mate-"3 rial have shown the following:
, .
- its high effectiveness for treatillg pyo-necrotic processes ' the curing period became 1.3-1,8 times shorter;

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20~22~

; - it~s high atraulnaticity: the f`orce of breaking the material off the wound surface is half as much;
; - a lower enzyme consumption rate: 10 to 30 times less as compared to that of native enzymeq used for treating the same :. ~
diseases; not less than 25 times as compared to the art-known analogues (in the analogues the enzyme content is 0.5 to 20~o~
whereas in the present therapeutic Inaterial the enzyme content is 0.02 to 0.5$3.
The stability tests of the present material a~ter its steri-~, lization ha~e shown that the present material preserves its curative effect after having been stored in packaged state , under normal conditions (i.e. at 20 C) for a period of at least 5 years, all of its physico-mechanical and sanitary-and-~' -hygien~c pr~perties being retained.
The present material has no side effects and contraindica-tions for its application.
~! The clini~al effectiveness of application of the present ~rlaterial possessing, a biological activity (for instance, con-~:l taining i~mnobilized hygrolytin3~ in comparison with the conven-; tional medicines traditionally used for treating pyo,-necrotic wounds in soft tissue~ is illustrated by the following data -` reported in Table 1 below-T A B L E

Characteristic of D u r a t i o n of t r e a t m e n t (d a y s) the treatment with the biologically with a native with a hyper-process active material acc~d- enzyme tonic salt so ~! ing to the invention tion containi '',' antiseptics '~ Complete purification '; of wounds an~ emergence ,~ of granulation islands 4.5 + 0.2 7.~ + o.5 9.5 + o.6 ... ... Table 1 to be continued ... ...
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, ... ... Table 1 continued ... ...

Complete healing of the wound from the be~innin~ of the treatment 15.0 + 2.1 20.0 + 1.5 26.0 ~ 2.2 ___ ______ The result~ of a clinic~l treatment have shown that the appli cation of textile materials containing immobilized enzymes for treating the patients affected with purulent surgical dis-eases of tissues makes it possible to shorten the total period of treatment by a factor of 1.3 to 1.8.
Enzymotherapy considerably reduces the period of beginning of purification of wounds and emergence of granulations: by a factor of 1.7 to 2.1.
The method for preparing the above-described material reside in the following:
A textile material whi~h is in itself chemicall~ inert,is subjected to the activation treatment to form therein reactive functional groups until their content reaches o.o625 to 3.125 mg-equiv. per gram~ of the carrier. The~ activation process depends on the chemical struature of a textile material (natu-ral, artificial, Rynthetie fibre~) and o~ its textile structur~
(weaving method used, thiclcness and number of thread folds, denstty of the textile material, etc.).
A cellulose- containin~ textile material is activated by subjecting it to oxidation with an alkali metal periodate~
such as, for instance, sodium periodate, to form cellulose dialdehyde. The activated textile material containg from o.o6z to 3.1Z5 mg-equiv. of reactive functional groups per gram of the carrier.

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20~ ~22a ~; Activation o* a te~tile Inaterial ;nade ~rom synthetic fibres (i.e. a material containing no cellulose), such as, for instanc-from polycaproami- fibre~" is carried out by subjecting a carrier-~aterial to hydrolysis with an acid, followed by ~ashin~
the carrier thus-hydrolyzed with water and by treating it with ~' Q solution of a co~pound containing reactive functional groups until the amount of the latter in the carrier reaches some , 0.0625 to 0.3125 mg-equiv. per gram of the carrier. It is advisable that strong mineral acids be used thereby, su~h as, . . ~
for instance, hydrochloric or sulphuric acids. It is advisable that the tre~tment o~ the hydrolyzed carrier made ~rom syntheti~
. .
fibres be carried out either by a solution of glutaric- aldehyde followed by washing with water, ar by a solution o~ dimethyl i sulphate-, followed by washing the carrier with ethyl alcohoI
;- and with a phosphate buffer solution.
~ Thereupon, the enzyme which is a biologically active prin-:?. ciple is immobilized on the activated textile material from buffer solutions having a pH ~f from 6.5 to 7.5 at room tem-!`:j , perature for 8 to 16 hours. Due to interaction between the reactive groups joined to the textile material and the free functional groups contained in the enzyme~, there is formed -~

~ between them a covalent chemiGal bond. As a result~ a textile `i material is produced, carrying a chemically (covalently) -~

~-` immobilized thereon biologically active substance having a ~;
-~ prolonged action. The immobilization process is continued ~` until a biologically active textile material containing some ;
~, 0.02 to 0.5~0 of the enzyme based on the weight of the material, is prepared.
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2~1~22~

The material obtaill~a in the form of cloth is squeezed out~
fiashed to remove the pilysically adsorbed enzyme, dried, and placed into a tight containe.r, for instance, sealed into poly-ethylene bags or paper-lined plastic packageR. Finally, the material i5 sterilized by the per se known methods, such aR, for instance, by irradiation or in a gaseous medium.
., ~ Prior to sealing into containers, the material, if need be~ j may be shape~ to assume a form.convenient for use, such a~, into napkins, lint, powder, etc., using the per se known methocl The. above-described biologically active material prepared by the above-disGlosed method may be used in a dressing means, The-latter has the form of a bandage compri~ing~ successively . arranged therein, a protective layer, an absorbing layer, !,~, and a wound-adjoining layer, at least one layer in this bandage ;~
. being formed from the above-deRcribed material. The. advantage ~, offered by the present dressing means over the art-known dress-ing.meanCf in the form of napkins made from similar biologically active materials resides. in the fact that the form:er ensures a fuller use of the therapeuti¢ efffefct of the immobilized enzyme. This fact ~ explained by the presence of an absorbing ~'~,3 layer which constantly suc:ks off splitting products, wound discharge and exudat:e, and which renders possible a longer and more effective use: of the biologically acti~e material constituting the wound-adjoining layer. For this reason, the . patient feel~ no discomfort caused by the need for frequent replacement of napkins wholly made from the biologically activf material~ such napkins being quickly saturated with splitting ~,, ' .
~','' ' .
., -- .

201~22~

products, ~ound discharge and exudate.
In another embodiment of the ~resent dressing means, the wound-adjoinin~ layer is made from the above-described biologi-cally active material~ wllile the absorbing layer is made from a textile ~oven, or non-woven, or l~nitted fabrie which does not contain any biologically aetive substanee.
! In a yet another embodiment of the present dressing means, the wound-adjoining layer is conYtituted by a biologically ~~ active material containing a proteolytie:~ or collagenolyti¢, :~ or bacteriolytie enzyme, while.the absorbing layer i.s also constituted by the same biologi¢ally active material containing a collagenolytic, or a bacteriolytic enzymes.
For instance~ if the wound-adjoining layer is made from ~ the material containing a proteolyti~ enzyme, then,. the absorb- :~
I ing layer makes use of a material containing a collagenolytie~
or a baeteriolytie enzyme. If the wound-adjoining layer employs a eollagenolytie enzyme, then, the absorbing layer employs I a ba¢teriolytie enzyme, and if the wound-adjoining layer uses ~:
;1 a bacteriolytie enzyme~ the absorbing layer uses a collageno-:l lytie enzym~
~' The present- dressing means ensures., simultaneously, splittin I of neerotic masses~ liquefaetion of viscous dense exudate~ and suppression of microbial growt.h in the wound.
An indi~idual. dressing means comprises elements intended to fix it on the patient's body, such as, for instance, an adhesi~e tape, a Velcro tape, etc.
. An indi~idual dressing means is prepared using conventional -. : .

., .

~:
~ -~2-:`` 20~ ~ 22~

technologies tradition~lly used in te-ctile and clotlling industr For better understanding of the essence of the present inven '~' ~ tion, attached herewith are specific ~xamples illustrative of - the present metllod for preparing a blologically active material .,j .
a dressing means, as well as of their application in practice.

EXA~PLE 1. Some 3.3 L of water were poured into a reactor, :
j its stirrer was set into rotation, and 9.4 g of iodic acid ', were added.
Into another reactor, the same amount of waber was poured,

3 a stirrer was set to rotate, and 1.6 g of sodium hydroxide were added. Both solutions were kept under stirring conditions ~` until complete dissolution of the crystals of the acid and sodium~ hydroxide during 5 to 15 minutes.
, Thereupon, both solutions were poured into a single reactor, .¦ stirred for 3 to 6 minutes to obtain a sodium periodate solutio having a pH of 5Ø
; 1 kg of medical cotton gauze was placed into the solution thus-prepared and held (activated) at room temperature for ;, 12 to 16 hours in the dark. Upon activation, the gauze was ` squeezed out, washed with 4 times x 10 L of water~ and squeezed out again.
As a result of the activation treatment, there are formed in the cotton gauze reacti~e functional groups, in the present case, aldehyde groups, in an amount of 0.0625 mg-equiv. per gram of the textile carrier.
~..

In order to prepare a phospllate buffer solution, two reactor were also used. One of them was filled with 3.3 L of water and, .`.j .

.

2~ 1 22~
, under stirring conditions, 5 g of potassium dihydrophosphate were added thereto, while 37 g of sodium hydrophospllate were added under stirring con~itions to the equal amount of water in the other reactor. Stirring was continued until the crystal~

. ~ ,, were totally dissolved for 10 to 20 minutes.
;~ Next, both solutions were poured to~ether into a single reactor to produce a phosphate buffer solution with a plI of 7.5 0.4 g of an enzyme, in the present cas~ hygrolytin, were added to the phosphate buffer solution thus-prepared under stirring ~onditions until the enzyme was cornpletely dissolved for 10 to 20 minutes. One kg of the activated cloth (i.e. ~;
dialdehyde cellulose) were placed into the reRulting solution and held at room temperature for 12 hours, during which period the enzyme imaobilization process takes place due to the emer-gence of covalent chemical bonds. The cloth was then washed until no protein traces were detected in the washings, whereupo ~-the cloth wa~ squeezed out and dried in ai~ for 24 hours. As a result, a biologically active material, in the present ca~e, cotton cloth featuring a prolonged therapeuti¢ action, is pro-ducad. The amount of the immobilized enzyme, in the present case~ hygrolytin, as determined by the enzyme loss f~om the solution, is 0.2 mg per gram of the textile cotton cloth, i.e.
accounts for 0.02dp of the weight of the cloth.
The dried material was shaped into medicinal forms, for instance, into napkins having the desired size, sealed into double polyethylene bags, and sterilized by ~-radiation usinF
the per se known procedures.

, .

_2L~_ 2 0112 h 3 L`~`IPL~ 2. A solution of sodi~Im perioclate was prepared following the procedure described in E~;ample 1, and 80 g of medical cotton gauze were placed into it. The process was further c~rried out lollowing the procedure of Example l to ob-,, .
` tain a cloth containing aldehyde groups in an amount of 0.78 mg-equiv. per gram of the textile cloth.
A phosphate buffer solutioI~ was prepared following the pro-cedure described in Example 1, and 0.24 g of nn enzyme, in ~ the present case, hygrolytin, were added thereto. The process 'i ~ was further carried out following the procedure of Example 1.
:, The amount of an immobilized enzyme~ i.e. hygrolytin, is equal to 1.25 mg of the enzyme per gram of the textile carrier (i.e. 0.125~o).
EXAMPLE 3. A solution of sodium periodate was prepared following the procedure described in E~ample 1, and 20 g of medical cotton gauze were plunged into it. The process was further carried out ~ollowing the procedure of Example 1 to obtain a cloth containing aldehyde groups i~ an amount of 3.125 mg-equi~. per ~ram of the cloth.
A phosphate buffer solution was prepared following thei proce dure of Example 1~ and 0.192 g of nn enzyme, in the present case, hygrolytin, were added thereto. The process was further carried out following the procedure of Example 1.
The amount of the immobilized enzyme, i.e. hygrolytin, was equal to 5 mg per gram of the cloth (i.e. 0~5~o)~
EXAMPLE 4. The process was carried out following the pro-cedure described in Example 1, with the ex~!eption that terrilyt i '~' 201~ 220 was used as an enzyme. The ammunt of the im.mobilized enzyme~
in the present case, terrilytin, accounted for 0,02/o based on the weight of the c.otton gauze.
EXAMPLE 5. The proces~ wa~ carried out following the pro-~, cedure described in Example 1, with the exception that used i as an enzyme was collagenase. The amount of the immobilized - enzyme, in the pregent case, colla~enase~ accounted for 0.02~o based on the weight of the cotton gauze.
EXAMPLE 6. The process was carried out following the pro-c.edure described in Example 2 ~ with the exception that terrilyti.

, was used as an enzyme~ The amount of the immobilized terrilytin ~...

accounted for 0~1 20a~ of the weight of the cotton gauze.
., : EXAMPLE 7. The proce~s was carried out following the proce-~ dure described in Example 2, with the exception that collagenas~
. was used as an enzym.e. The amount of the immobilized collage- ~:
nase accounted for 0.140$ of the weight of the textile; gauze. :
E~AMPL~ 8. The process was carried out following the pro- .
cedure described in Example 3, with the exception that terri- :
lytin was used as a~l enzyme. The amount of the.immobilized terrilytin accounted for 0.5~,~ based on the weight of the cotton gauze.
EXAMPLE ~. The proce~s was carried out following the proce-. dure described in Example 3~ with the exception that collagenas .
`- was used as an enzyme. The amount of the immobilized collagenas . .
accounted for 0.5~ based on the weight of the gauze.
A~IPLE 10. The process was ~arried out following the ~. -procedure described in Example 1., with t.he exception of the fa~ ~ .

' .

, -, .
'~
~.

2~ ~22~
, that, as said texti].e material, use was made of a gauze from cotton-and-viscose fibres.
E~Ai~IPLE 11. Th~ proces3 was carried out following the ~! procedure described in ~xample 2, with the exception tha~, as i -~ said textile material, use was made of a gauze made from cotton ; -and-viscose fibres.
EXAMPLE 12. The process was carried out following the pro-cedure described in Example 3, with the exception that~ as said textile carrier, use was made. of a gauze ma~e from cotton-and--vlscose fibres. ~-EXAMPLE 1~. The process was carried out following the procedure described in Example 1, with the exception that for i preparing a buffer solution, 20.7 g of potassium dihydrophospha and 11.~ g of sodium hydrophosphate were added to obtain a phosphate buffer solution with a pll of 6.5.
EXAMPLE 14. The process was: carried out following the pro-.~1 cedure described in Example 1, with the exception that for ~ preparing a buffer solution, 11.9 g of potassium dihydrophos-:~ phate and 23.. 6 g of sodium hydrophosphate were added to obtain -l a buffer pho3phate solution with a pH of 7.
E~AMPLE 15. The pro~ess was carried out following the pro-~ cedure described in Example 1, with the exception that trypsin . was used as an enzyme.
EXAMPLE 16. The process was carried out following the pro-~ c.edure de3cribed in ~xample 1, with the exception that use :~

`! was made of a phosphate-citrate buffer solution having a pH ~.
1 of 7.5. Said solution was prepared using the following procedu ~

,,, . :-: .

'`1 ' '~" ~;~:

~ -27_ :
Some 1.8 L of water were r~oured into the fir~t reactor, wher '~ after 19.06 g of citric acid monohydrate l~ere added thereto under ~tirrn~ conditioni3, whereas ~ome 8.25 L o~ water were poured into the gecond reactor, followed by addition of 329.3 g sodium hydrophosphate under stirring condi'ti.on~. Stirring was continued until the complete~-aissolution was accompliished, whereupon both 901ution8 were poured together.
The process wai~ further carried.out ~ollcwing the procedure '. of Example 1.
EXAMPLE 17.'T~q-proc.e~g was carried out following the proce dure deicribed i'n Example 13,. with the exception of the fact ~'~' that use was made of a phosphate-citrate bu~fer siolution. In .- order to pr~pare: ~aid solution, 71'.7 ~ o~ ci'tric acia mono-hyd- :
-:` ratQ were. add'~d to 1.8 L of water poured into olle reactor under ' stirrin~, while 224.. 5 g of sodium hydrophosphate were added' ., to. 8.Z5 L o~ the water poured' into the other react:~r, also .~ under ~t.irring. Upon complete s.olubilization of the sal~.s, the ',''':' solut:ions thus.-obtained: were pu~ together to obtain a buffer ~ solution ha~ing a pH = 6.5. The process was further carried .,;~
, out following the procedure of' ED~mple: 13 .
., XAMPLE 18. ~h~-pro¢ess wa9 carried out following the pro-', cedure de~cribed in Example 1~4 ~ with the exception of the fact that, in order to prepare 'a phosphate-Gitrate buffer solution, ' 38.1 g o~ citric acid ~onohydrate. were added under stirring .~................ condî'tionis to-1.8 L o~ wAter poured into one-reactor, while-~:'! 290.5 ~ of sodiu~i hydrophoi~phate: were added to 8.25 L of water ~

,~! contained in the other reactor. A~ter the solubilization o~ ~ :

~:, ,~, :

,: ~ ~' : .': , . ' ' : : : ~ :

20~2~

tlle salts,both ~olution~ were put to~ot!~er to yield ~ buffer solution llaving a pH = 7. The proceg~ wa~ ~urther carried out following the procedure ~f Example 14.
EyA~prJE 19. Some 50 L of 3N-~Iydrochloric aGid were poured into a reactor, and 1 kg of a knitted fabric ~ade from poly-caproa~ide fibre~ were placed into it. Thereafter~ the acid temperature was c~u~ed to rise up to 60 C, and the ~abric was held at thi~.temperature~-~or 4 hours. Upon termination o~ the hydrolysls, excess. hydrochloric acid l~a~ cooled down an~ dis-earde~. The,knitted fabric was washed with water until no hydro-chloric ac-id traces were dete.cted- in the washi~gs, Then, some 40 L o~ a 5~-~,lutaric aldehyde were poured into the reactor, and 1 kg of the ~nitted fabri~ sub~ected to hydrolysis were placed into it... The temperature o~ the:glutarie aldehyde 901u-tlon.was brou~ht to 50 C and maintained at this: le~el ~or 4 hours. Then, the glutar~ aldehyde solution was c.ooled down and discarded~ The fabric was wa~hed with water until no smell .~.
~ of glutaric aldehyd~ was le~t. The,fabri~ acti~ation proces~
.
-~' wa~ thus terminated.
As a result of the activatio~ txeatment, there are formed ,.
, in the knitted fabri.~ reactive func.tional groups, in the pregent ', case, aldehyde groups, aontaining 0.. 0625 mg-equi~ per gram~
'. o~'the textlle fabriG..... ' ' Preparati.on of the phoiphate.-buffer solution was carried :-,:
! out ~ollowin~ the proeedure o~'Example l. For this purpose, som~

: o.66 g Or an enzyme~ in the present case, terrilytin, were .~ added.to the phosphate buPfer ~olution thuY-obtained at a pI~=7.

~., . :
. . .
:

~. -29-20~ ~22~
~. . .
Stirrin~ was continued until complete qolubilization within 10 to 20 minutes.
1 kg of the activated lcnitted fabric made from polycaproamid fibre~ were placed illtO the thus-prepared terrilytin solution in the phosphats buff0r solution having a volume of 6.6 L and an enzy~e concentration of 0.01%, and held at room temperature for 9 ~ours. There took place the enzyme immobilization process due to the formation of covalent chemical bonds. Thereupon, the fabric-wa~. washed with a physiologic salt soIution and water u~til no enzy~e (protein) traces. were detactable in the washing~s, the a~ount of the enzyme (protein) being controll~
by the per se known procedures.
The fabric thus-treated was then ~.queezed out and dried in ! ~
3 air for 24 hours to obtai-n a biolo~ically active material, in ¦ the pre~ent case, a knitted polycaproamiae fabri~, featuring a ., .
~ prolonged`therapeutic ac.tion ~ The material thus-obtained oontains 0.02~ of im~obilized ;l ~errilytin ~i.e.. 0.2 mg of the enzyme per gram of the carrier).

Af*er drying~ the material was shape~ into medicinal forms, ¦ such as~ for instancel into napkin~ ha~ing a de~ired size, ealed into a container, and: finally sterilized in a ga~eous .I mediu~ using the per SQ known procedure~

EXAMP~E 20. An activated knittea polycaproamide fabric .

.3 was prepared followlng the procedure described ln E~a~ple 19, .

-~ wlth the exception that use was ~ade of a 10~p-glutaric. aldehyde :
~:5 After acti~ation, the content of aldehyde groups in the fabric .`! was 0.155 mg-equiY. per gram.of the fabric.
;., . ~

-3~~
20~1 2~

1 k~ of the activated fabric were placed into a reactor c.ontaining 6.6 L of an 0.03/~-terrilytin solution in a phosphate buffer solution having a pH = 7.5, and held at room temperature . for 8 hours, Further~ the fabric was subJected to treatment described in Example 1g. The biolo~ically acti~e material ~' thus-obtained contal~s o.6 mg of tsrrilytin per ~ra~ o~ the fabric, i.e. o.o6dO based on the wei~ht o~ the fabric. The pro-cess was continued following the procedure of Example 19.
. EXA~PL~ 21~. An acti~ated knitted po-lycaproamide ~abric ~ wa~ prepared following the proc.edure described` t'n Example 1-g, : with the e~ception o~ the fact that ~ 15~-ælutari'c aldehyde was usod. ~ter acti~ation, the content o~ aldehyde group3 ~` in the fabric was 0.26 mg-equi~. per gram o~ the fabric.
1 kg of the acti~ated: fabric were placed into 6.6 g of an ;' 0.1~0-terrilytin solution in a phosphate bu~fer ~olution having ;.
,' a pH of'7.5, and held at roo~ temperature for 10 hours. Furthe~:! . . .
processin~ of the fabric was carried out following thff pro~e.

' dure.of Example 19.

The. resulting materi`al containg.2 mg of t'errilyt'in per gram ~' ~! of the carrier, i.e. 0.2~ of the enzyme basëd.on the weight ~`' o~ the fabric-~'"' EXAMPLE 22-. The process was carried out. followi'ng the. :

~ procedùre described in Example 19, with the exception o~ the f; ~:

'. that, as an enzyme~ use was ~nade of trypsin. The material ::~

thus-preparea cont'ains 0.02~o 0~ immobilized trypsin on.the te~ :

~ tile carrier, in the present case., on the knitted fabri'c from '.. ~ polycaproamide fibres.
., ' . ~:

~,'- ':, : -31-2~ ~22~
. - .

~ XA~PLE 2~. The process was carried out -fol:Lowlng the proce-dure described in ~xample, with the sole e~ception that~ as an enzyme, uge wa~ made of trypsin, and 1 kg of the activated knitted polyc~proamdde ~abric were placed into 6.6 L of an 0.03~-tryp~in solution in a phosphate buffer solution and held at room temperature for 9 hours. The~process was further conti-nued~ folIowing the procedure af Example 20.
. The material thus-prepared: contained o.o6~ of trypsin based :1 on the weight of the knitted polycaproamide fabric.
.~
.. 1 . E~AMPLE 24. The.pro~ess was ¢arried out following the pro-cedure described in Example 212 w~t.h the sole exception of the fact that the immobilization treatment was carried out in an .l 0.15~-trypsin solution in a phosphate bu~fer solutio~
The: material thus-prepared contains 0.3~ of trypsi~ based .l on the weight of the fabric.
~1 EXAMPLE 25. A polycaprQamide knitted fabric was held in ~..
a reactor filled with dimethyl sulphate at a bat.~ modulus of ~ 10 for 10 hours at roo~ temperature. The.excess dim.ethyl sul- .
`j phate.was poured out, whereafter the ~abri~ was~washe~ onee with et.hyl alcohol at a bath modulus of 20 at a temperature :~ :
of 4 Ct followed by washing t.hrice with a phosphate buffer ~:
solution ha~ing a pH of 7.5 a~ a te~perature of ~ C. The fab-ric thus-processed c.ontained reactive imidoethereàl ~rroups.
The immobilization treatment of the enzyme, in the present L .
case, o~ trypsin, W~9 carrïed out. at a bath modulus cf 10 ~rom an enzyme solution in ~ phosphate buffer solution having a concentration af o.o4$ for 16 hours at room temperature.

,:~ "
.

2 0 ~ 1 2 2 ~

~ h~ resulting ~aterial contains 0.2~;~ of the im~Lobilize~
trypsin, i.e. 2 m~ per gra~ o~ the carrier.
The fabric was then washed wit'h a physiologic salt solution and water until the washings contained no enzyme traces whose amou~lt was controIled by the per se known procedures.
Thereupon~ the fabric was squeezed out and: dried in air for 24 hours t-o yield a ~aterial, in the present case, a Xnitt~
,' polycaproamide.fabri~.~ featuring a biolo~ical acti~ity and a prolon~ed therapeutic.action.
,' The~ ~aterial thus-prepared, after being drie.d, was shaped ,~ .
`,," into ~edicinal for~ns, su~h as, for instance,. into.napkins havin~ , -, a desired size, sealed into double-waIled poIyethylene bags, '. and sterilized by gamma-rays using ~he per se k~own pro.c.edures.
~ EXA~IPLE 26~ The process was:carried out followin~ the proce~
dure dc~c~ibed in Example 2~, wlth t'he sole eYception of'the fac~t that the immobilization treatment of trypsin, used a~ an :~
.", enzyme, was carri'ed'out in an 0.008%-trypsin solutïo~ i'n a ~ - phosphat'e buffer solution at a bath m.odulus of iO for. 12 hours~
::~ The:resuIting materi'al c:ontaine~ O.. Q4~ of the immobilize~ tryp-;`~ sin, i.. e.. 0.4 mg per gram of the carrier ,, XAMPLE 27... The process was carrIed out following the, prQCe dure described in Example Z5, wi'th the:sole excepti~n of the- :~
` ~aGt. that the immobilizatio-n treatment of trypsi~ was. performed ,. in an 0.024~-trypsin solution in a. phosphate:buffer solution :.~ at- a bath modulus of 10 for 10 hours. The.material thus-obtaine ,. c:ontained 1.2 mg of trypsin per ~ram of the carrier, i.e. 0.12~,'~. ,.
of trypsin per gram of the carrier.

;"', :

` 33 2 0 ~ 1 rJ 2 3 ~ XAMPL~ 28. The process was carried out following the proce dure describe5d ia Example 19, with the exception that r a~ an enzyme, use wa~ ~ade of l~sozyme. The resulting material contai 0 . 02~o by weight of immobiliz~d lysozyme.
FXA.~LE Z9. The process was carried out following the proce dure de~c-ibed in E~asnpLe 20, with the exception that, as an enzyme, u~e was made of lysozyme. The resulting material contai ed o~o6~ by weight of immobilized lysozyme.
EXAMPLE 30. The pro~ess was carried out following the proce-dure described in Example Z1, wïth the e~ception that, a~ an enzyme, use wa~ made o~ lysozyme.The materiaI thus-prepared contained 0.2~ by weight of immobilized lysozyme.
I EXAMPLE ~1. The proce~s was carried ou~ following the pro-~'i :
cedure de~cribed in Example 19, with the exception that, as an j enzyme, u~e was made o~ protosubtilin. The resulting material a o.a2~ by weight of immobilized protosubtilin.
EXAMPLE ~2 . The proce~s wa~ ¢arried out followi~g the pro-cedure described in Example 20, with the e~cept`ion that, as an enzyme, use was made o~ protosubtilin. The mate~ial thus-prepa-red containe~ o.o6~ by weigllt of immobilized pro$osubtilin.
EXAMPLE 3~. The proces~ wasi ~ar~ied out following the pro-cedure de~cribed in Exa~pIe 21, with the exc~eption that~ as an enzyme, use was made of protosubtilin to obtain a material containing 0.2$ by weight of immobilized protosubtilin.
E AMPLE ~4 . The steriIe, biologically acti~e material, pre-' pared following the procedure of Example 1, is a wo~en, medica]
-~, grade ? cotton gauze containing, immobilized thereon and co~alel ;, , ~

`"'I ;~
:.

_3L~_ 2~1122~

ly bound thereto, hygrolytin. The material contain~q 0.2 mg o~
hy~rolytin per ~ram of the woven cotton ~auze.
The material is turned out in the -form of medical napkins and may be used for treatment of abscesses, phle~ons, postoper tive compliGations af a surgical suture.
The uqie of this biologically active material ensure~ a pro-longed thera~eutic effect of an enzyme for 48 to 96 hours, wherea~ the life perio~ o~ a native enzyme which is used for treatment of the ~ame aisQase~, iq not more than 1 hour. After a therapeutic application of the claimed biologi¢ally acti~e material, a complete healin~ of a pyo-necrotic wound on the -soft tissue~ of a le~ is obser~ed after 17 days. In some ~ases, during the first hours aft~r applying a bandage, the patient~
experienced a gentle feelin~ of burning, pricking of the wound surface under the bandage. No allergi~ reactions of the patient organiqms~ were obserYed~
It has been concluded that there exists a covalent type of a bond between the carrier and the-enzyme; of the claimed material~ proceeding ~rom the fact that after treating the material with salt solutions, such as~ for instance~ with sodium chloride, or with buffer solutions, such as, for instanc~
wit~h a pho~phate buffer ~olutionf the claimed material still retained its enzymatic activity. In the course of such treat- -~ment~ physiGally ad~orbed proteins ~enzymes) are washed out of the carrier.
After five years of storin~ under normal conditions, the claimed ~aterial still retained i~s therapeutic properties.

~ .

. ~ , ~ 3 - ~
',tj ~

~35-201122~

: EXAMPL~ ~5. The~ bio10r,~ically actl~e material prepared in : ..
accordance with ~,xa~nple 1'had a composition identical to that of Example 1, except that It contained 1..25 m~ o~ hygrolytin per gram of woven cotton ~auze. T~e.therap~utic application of thi4 wo~en cotton ~auze c.ontainin~ hygrolytin, immobllize~.
thereon and' ~ovalent.ly bound thereto., produced a prolonged ' therapeutic ac.tion which lasted some 48 to ~6 hours as compared :
,t! t:o an hour-long period of action of the natiYe enzyme,.
~ij .
.-', After apply~ng thi.s biologically acti~e material, a complete :~
healing, of a pyo-nec~toï¢.wound in the 50ft. tissues of a leg wa~ obser~ed a~ter 15 days~ the other-results being a~lalogous , to those of Example 34. :
, EXAMPL~ 36. The bïologically acti~e material prepared in '~ accordance with Example 1' had a GompOsitiOn ldentical to that ~.
~3 of Example 1:, exeept, that it containe~ 0.5 mg o~ hy~rolytin ~:
`'3 per gram of a woven ~i.otton gauze,. This material was applie~
`, in accordance with Example 24. After applieation of this biolo-.,-. gically actiYe material, a complete healing o~ the wound ~, was obserYed after 14 days, the.other re~ults being analo~ous ~ to those of`Example 31l. ~
.~ EXAMpT,~ 3'7. The~biologically aotive materia? prepared ~:
in accordanGe with Example 1 had a compositi`on identicàl to I that of ExampIe 1', except'that used as a textile carrier was `1 a knitted.medical cotton gauze~ The results of the dedical ~.
''.¦ application of this material are the same as those of Example ~
..... .
~ EXAMPLE' ~8. The biolo~i¢ally acti~e material prepared in 7`'~' accorda~c-e with Example 1 had a composition identicaI to that .~. 1 .

., .
:'1 2~1~ 22~

, of ~ample 1, e~cept t~lat u~ed as a textile carrier was a non-~ woven ~ate~R~ fabric ~rom. cotton fibres. The reslllts Or medica .~ ~pplication ~ this material are the ~ame as those of ~xample 3 EXAMPLE ~., The biologically active material prepared in accordance with Example 1 had a compositi'on identical to that of Example 1', e~cept that used as a textile carrier was a wo~en .
, linen fabric-. The results of medical applicatIon of this mate-,, rial are the sam,e as those of'Example 34~
E A~LE 40.. The biologically acti~e material prepared i`n accordance with Example 1' had a cQmposition identical to that . o~ Example 1:, except t.hat u~ed as a textile carrier was a wo~en natural silk fabri~. The results of therapeutic.applica-tlon of this material are the same as those o.f Example 34.
EXAMPL~ 41.~ The biologically a~ti~e material prepared i~
' ac.cordance with ~xample 1'9 had a composition identicai to that '.~ of`Example 19', except tha~ used as a textile carrie~ was a :~
.. '' knitted' polycaproamide fabric.
~;! "" The results of the medical application of this material are '.
,, the same a~ those o~ Example 34.
~ EXAMPLE 42.~ The biologic.ally active material prepare.d in `~. accordanG,e with Example 19 had a compo~ition identlcal to.that of Example 1~, except that used' as a textile carrier~waR, a ''~ knitte~ polyurethane fabric~
~' The results o~ the medical application of this material wer~
. ` .
"~ the ~ame as those of Example 34., Example 4~. The,biologlcally acti~e material prepared in '~
accordance with Example 1 had a Gomposition identical to that , .. . .

: -37-2 ~ 2 ~

of ~xample 1, except that used as a textile carrier was a knitt( viscose fabric.
The results of the medical application of this material were the same as those of ~ample 34.
EXAMPLE 44.. The biologically active material prepared in accordance witll Example 1 had a composition identical to that Example 1, exeept that used as a textile carrier was a secondary acetate cellulose knitted fabric.
The results of the medical application of this material were the same as those. o~ Example 34.
E~AMPLE 45, The biologicalIy active material prepared in accordance with Example 21 had a composition identical to that of Example 21, except that it contained lysozyme. This material contained 2 mg o~ lysozyme per gram of-the textile: carrier.
The use of this material containing immobilized lysozyme covalently bound to the: earrier~ produced a prolon~ed therapeut-~ action which lasted somc 48 to 96 hours against o~e hour-long :i per~od o~ therapeutic a~tion of the native enzyme.
; After a medlcal application of this material, a c.omplete healing of a pyo-necrotio wound in the.soft tissue~ was observe~
after 15 days.
Treatment of purulent. wounds at their hydration phase with a textile materiaI containing immobilized lysozyme, shorten~
the period of elimination of the wound infection, cuts ~hort - the alternative purulent inflammation, promotes the purificatio-of wounds from pyo-ne¢rotic tissues~ stimulates the synthesis . of glycogen and nucleic acids (DrJA and:RNA) in the woun~ cells, i .
~,~

~r ., -38- , ~ 201122 .~ .

acti~ateg the proliferation of co~necti~e tissue cell~ (fibro-. blasts), accelerates collagenosis, the contraction of wound ', . edges~ the contraction and epithelization of the wound.
., $XA~IPLE 46. The,biologically ac,ti~e material prepared in "-, accordance with Example ~ ad a compositio~ identical to that of Example 5.. This material c.ontained a collagenolytïc enzyme, ... namely, collagenase, in an ainoun~. of 0~2 ~g of colla~enase, ' per ~ram of the textile carrier~
~, After a medical appli¢ation of this materi'aL, a complete : healing of the wound was observed after 16 days,~ the other ,~. results o~ the medical appli~ation of this material being ~.
analogous to those of Exa~ple~ 34 and.45., F~AMPLE 4:7,, The biologically active material,prepared in ~, accordance with Example 1"5 had a compositi`ou identical t~ that ' o~ Example 1 This ~at,erial was turned out in the form of lint having ;~ fibres 1.0 mm long and was,u~able for treatment of purulent, deepS punctured wounds.
~- The therapeuti,c effect, nameIy, a complete heali'ng of wounds ,: . . .~ was ob~,erved a~ter 17 dàys.
.. EXAMPLE 48. The.biologically active material prepared in accordan~.e wi`th Example 15 had a compositi,on identical to that of Example 1'. ~:
This material was in the form of lint haYing fibres '.

~,i 8.0 mm long, and it was used to accelerate the treatment of .~`I, dental infection foci, such ag, ~or instance, chronic granulate periodonti~is.. This material in the form of lint produGes an ''`3 :

, `. . :

2~22~

antie~u~ative effect~ i~ acti~ely promotes the outflow oP exuda through ~ tooth root channel ~even through curved, narrow, per-meable channelsj.
E~MPLE 49. The biologically activ~ material prepared in accordance with E~ample 15 had a composition identical to that of ~xample 1.
The material thus-obtained was in the for~ o~ lint having fibres 15 mm long, and it was used for treating an inflam~latory process in the reGtum.
The therapeutic effect was observed. aft.er 16.5 days.
E AMPLE 50. Th~ blologically aGtive material prepared in ac¢ordance with Exa~pie 15 had a~composition identical to that !
of Example 1.
This material wa~ in the for~ of a powder Kaving a particle size of 0.001 mm. It was used for treatin~ postoperative comp]i ~t cations of surgical ~u~ures.
After a medical applic.~t of this powde~ possessing a ~.i biological, includin~, an enzymatic activity, a complete healir ~ of the sut.ure was obser.ved after 17 days.
EXAMPLE ~. The- biologically active material prepared in accordance with Example 15 had a composition identical to that.
.~ .
~ of ~xampl.e 1.

: This material was in the form of a powder having a particle 1 ~ize of 0.5 m~. It was used for treating pyo-necrotic processe .. of complicated localizations and difficult-of-aGcess cavities.
, ~

The therapeutic effect was observed after 17 days.

.A.~PL~ 52. The-biologically active material prepared in ~, , .s:,:. . ..

- l~o~
201~22~

`: accordance with Example 15 had a com?osi tion identical to that of ~xample 1 .
. This material was in the form pf a powder having a particle size of 1.0 m~, and it wa~ used for treating phleg~ons.
. Tlie therapeutic effect was observed after 16.5 days, EXAMPLE 53. A dressing means in the form of a bandage consisting of three layers and a fixation member made in the ' form of an adhesive tape. The.bandage cQntained, arranged in succession therein, a protective layer, an absorbing laye-r, and a wound-adjoining layer..
~; The protective layer was made of a woven cotton gauze, while the absorbing layer was made of three strata of a woven cotton gau~e. The wound-adjoi~ing layer was also made o~ a woven .! cotton ~auze: containing a co~alently bound:thereto, and immobi-lized thereon~ proteo-lytic enzyme, namely, trypsin in an amount of~ 0.02 wt.~
!~
ll of the above-mentioned.layers were laïd, in suc~ession, one on top the other, tailored to form: napkins having a desirec ~.
size~ such as, for instance, 150 x 200 mm2 or 100 x 150 mm ~ :

interconnec:ted by some sewing method~ the fixation member ~I being atta¢hed to the protective layer. The dressing means . thus-prepared was ~ealed into a c.ontainer and sterilized by i~`;; exposin~ it to gamma-radiation,~ whereupon the dresi~g means became ready for application and storing.
EXAMPLE 54~ The dressing means was ~ade in ac¢ordance with Example 53 and contained an immobilized enzyme in an amou of 0.125 wt.%.

. ~ .
,~ , ' ' :
- ::

,.i :: :

20~ 122~

E~A~I~L~ ~5. The (lr~ssing mcans was pr~pared in accordance with E~ample 53 and contained an immobilized enzyme in an amount of 0.5 wt,',l.
E'AMPLE 56. The dres3ing means was prepared in accordance with ~xample 53 and contained an im:nobilized collagenolytic enzyme, namely, collagenase, in an amount of 0~02 wt.~p.
XAMPLE 57. The. dressing means was prepa~ed in accordanee with Example 53 and contained a Golla~enolyti~ enzyme an amount of 0.140 wt.%.

, EXAMPLE 58.The dressing means was prepared i~ accordance with Example 53 and contained a ~ollagenolytie enzyme i~ an ` amount of 0.5 wt.~p~
. EXAMPLE 59. A dressing means in the form of a bandage c.o~-- sisting o~ three layers and a fiYatio~ me~ber c.ontriv:ed as an ~`. adhesi~e tape. The band~ge~ containe~,. suc~essively arranged therei~,. a prot:eet:i~e laye.~, an absorbing layer, and.a.woun*~
~ adjoining layer. The proteGti~e layo~ was made frQm a knitted Q''`,~ '` textile fabric made from polycaproamide fibres. The: absorbing layer waa.made ~rom four strata of a knitted vi.sc.ose fabric.
The wound-adjoining layer was. prepared from a knitted;poly-eaproamide fabri~ containing. a co~alently immobilized bacterio-lytie enzyme, ~iz. lysozyme, pre~ent. in an amou~t of 0.02~ by weight.. Ths abov.e-specified layers were arranged in the claimed dressing means in a manner identi~l to that of Example 5~. :
! EXAMPLE 60~ The dressing mean~ was prepared in accordanee 1~. '`
lS~ with ~xample 59 and had a lysozyme content of 0.12~ wt.,~.
..~j X~PL~ 61. The dressing mean~ was prepared in.accordance ` 'i f ,' -, ,: ' "
' ~', . ^ ! ~: , . .
: .

2(~22~

with Example 59 and contained lysozyllle isl an alDount of 0.5 by weight.
XAMPLE 62. A dressing means in.the form of a bandage con-sisting of three-layers and a Pixation member contri~ed as an adhesive tape. The bandare contained, arranged in sucGession therein, a protecti~e layer, an absorbin~ layer, and a wound-adjoining layer~
The protective layer was formed from a non-woven glued hydro phobic ~abric, while the absorbing layer was formed from three strata of a non-woven glued hydrophilic fabric. The wound-adjoining layer is formed from a single layer of`a knitted textile fabric made from polycaproamide fibres and containing : a covalentIy im~nobilized proteolytic enzyme, na~ely, trypsin, in an amount of O.O~ wt .$.
The formati.on o~ the present dressing maans from the above-enumerated layers followed~ the procedure of Example 53.
. F~-AMP~E 6~ The drcssing means was prepared in accordance with ExampIe 62 and contained trypsin in an a~ount of 0.126 by weight.
;, EXAMPLE 64~ The dressing means was prepared in accordan~e .~ with.Example 62 and contained trypsin in an amount of 0.5 wt.~.
~ XAMPLE 65. A dressing means in the form of a bandage consisting of three layers and a fixation member formed as an - adhesive tape. The bandage contained, successively arranged therein, a protective layert an absorbing layer, and a wound~
adjoining layer.
~, The protecti~e layer was made from a wo~en cotton gauze, wh , ~

. . . ~ .

20ll22a the absorbing layer Wa3 made from cotton lint containing colla-gonase on an a.~iount o~ 0.02',~ by weight and con~isting of flbres :, -~, . 1.0 mm long. Thc lint layer was dispo3ed on the interhal side , of the,protecti~e layer of the bandage. The wound-adjoining .~' layer was prepared from a wo~en cotton gauze c.ontaining co~alen ,~. . , ly immobilized hygroIytin an amount of 0.02~o by weight. The ,~ above-mentioned fabrics are laid one on top the other in such ~,, ,.-. a manner as to contain lint interlayers between the main layers ',~ whereafter the whole bandage i.s tailored into napkins. The . . .
, , pro~edure of formi~g a dressin~ means containing such layers, was identical to that of Example 53. :~
, , FXAMPL~ 66~ A dress,ing meansi was, prepared in accordanc,e .
'~,; with Example 65, exc.ept that its~ absorbing laye~ was made ~ ' from a cotton lint containing lysozyme in an amount of 0.126~J

'~'' by wei~h~ and ha~ing fibres 10 min long~
..
EXAMPTE 67. A dressing means was. prepared. i~ accordance with E~amplè 6$,~ except that its-absorbing layer was made from a Gotton-~isco~e li~t contalning lysozymeinan amount of 0.5%
~, ,', by weight and haYing ~ibres 15 mm lon~
~ EXAMPLE 68.. A dressing mean~ was prepared in accordance.

,~ ' with E~ample 651 except.that its absorbing Layer was made from .~ pulverulent, collagenase.in an amount: of 0.2% by weight and with .~ a particle size of OrOOl mm~
;, ~YAMPL~ 69. A dressing means was prepare~ in accordance with Example 65, except that its absorbing layer was made from a pulverulent. collagenase.in an amount of 005~p by weigh~ and '~'~
'.' with a particle size of 0.1 mm.

. ' ' .

,, `
r ~ r ' . .A . ~ ' ' ' ' ' ~ ' ' _1~4 _ - 2~ 22~

r~XA`IPrE 70. A dressing means was prepared in accordance with Examplo 65, except that its absorbin~ layer was made from a pulverulent lysoz~me in an amount of o.5~10 by weight and with~ a particle size of 1.0 mm.
EXAMPLE 71. A dressing means prepared in accordance with Example 53 was applied at the sta6e of pre-medieal aid, just after infliction of a trauma of soft tissues in the gluteal regin, havin~ a size of 6 x 4 cm and 4 cm deep. Upon expira-tion of 12 hours after infliction of the trauma, no oedema and perifocal inflammation were observed; the internal surface of the wound was clea~, and the ti~sue~ were ~iable. The comple healing of the wound was observed after the sur~icaI treatment in 14 days.
EXAMPLE 72. A dressing means prepared in accordance with E~
ple 54 was applied at the stage of pre-medical aid for treating a trauma on the hi;p, havin~ a ~ize o~ 3 X 4 cm and about 3 cm :, deep. When 10 hours later the patient was admitted to the clini , no oedema and perifocal inflammation were obser~ed, and the tissues were viable~ The complete healing of the wound a~ter the surgical treatment was observed after 14 days.
XAMPLE ~3. A dressing means prepared in acoordan~e with Example 55 wa~ applied at the stage of pre-medical aid fo~
treatlng a trauma o~ soft tissues, having a size of 5 x 2 cm and a depth of up to 5 cm. When 11 hour~ later the patient was admitted to the clinic, no oedema and perifocal inflammati were observed; the internal surface of the wound waR: clean, : ~:

and the tissueY were viable. The complete healing of the wound 20~ 2~
was observed aft~r the surgical treatment in 13 day3.
EXAMPL$ 74.. A dres~ing meanA was prepared in accordance with ~xampl~ 53. Its wound-adjoilling layer was made from a materiaI containing, immobilized thereon, trypsiin in an amount of G ~2~o br weight, while its absorbi~g layer was made fro~.a ~ material Gontaining immobilized lysozyme in an a~ount of O.Z.
'~ by weight.. All of the layers o~ the bandage were in the for~
`' of napkins.
' Said dressing means wa~ applied. at the stage of pre-medical !, aid for treating a trauma inflictsd to the soft Ieg tissues ii~ and having a 9izc of 5 x 5 cm and a depth o$ 3 cm. When 9-hour~ later after in~liction.of the trauma, the patient was taken to the cliniG, the internal surface of the wound was clean and no inflammatory effects were observed.
After the surgical treatment:" a complete healing of the ~ wound was observed in 12 daya.
s ~XAMPLE 75. A dreqsinæ meano prepared in accordance with x Example 74 and having an absorbing layer constituted by a stratum of lint'~ wa~ applied at the stage of pre-medicaI.ai~
for treating a trauma i'nflicted to the soft l'eg tissue~ and having ~ size of 6 ~ 3 cm and a depth o.f 3 Gm. The patient was admitted'to the clini~ 7 hours later, and a complete~
healing of the wound after its surgical treatment was obger~ed in 12 days.
EXAMPLE 76. A dres3ing means prepared in accordancs wit.h ~xa~ple 74, except that its absorbing layer was made of lint, was: applied under conditions identical to thoge of Example 7ll ' , .' .`,''`

-l~6-20~22~

A complete healing of the wound was obs:~rved after 11.5 day3.
,~AMPI,E 77. A dressin~ me.ans was prepared in accordance with ';`xample 53. Its wound-~dJoining layer was made from a material containing imMobllized colla~enase in an amount of 0.2',~ by weight, while its absorbin~ layer was made from a ~aterial containing immobilized lyso~y~e in an amount of 0.3 by wei~ht.
This dressing means was applied at the stage of pre-medical aid for treatin~ a trauma inflic.ted to the soft leg ti~sues and having a size of 3 x 3 cm and a depth of 2 cm. When 14 ~.
hours later the patient was admitted to the clini'c, the interna.

surface of the wound was.clean. A complete healing of the wound after it,s sur~ical treatment. was observed in 12 days. ~ :
~XAMPLE 78. The process was conducte~ in accordance with Example 1, except that for o~idation use was made of a potassiu hydroxide solution and the activatio~ treatment was carried. ~:~
, ~
out for 16 hours. The material thus-prepared c.ontained an .

'.' immobilized enzyme in an amou~t of 0.02 wt.~. '.-~

~' EXAMPLE 7~. The process was conducted in accordance with :

., ~xample 3, except:that the duration of the~enzyme immobilixatio ' .~, treatmen*.was 16 hours~ The material thus-prepared~was subjecte to additional mechanical treatment. in a mlll and turned out ~ ~

i~ the form of lint consisting o~ fibres 15 mm long. The enæyme ~ .

~ conte~t in this material was 0.5 wt.~p.

.. ~. F~AMPLE 80.. The proce~s wag c~rried: out in accordance with -~

'~ ~,xample 12, except that the duration of the enzyme immobili2ati ` treatment; was 16 hours and the final material was additionally ~ -~ .

: .
.:~

, ., . ~ .
' , ; ~ . . . ~ . . . ': . ` , , . ' ' !

:.
-~7-20~ 122a subjected to mechanical treatment in a mill to yield a powder having a particle size of ~..5 mm. The matorial contained 0.5$
by weight of an enzyme.
The material thu~-prepared was placed into a container by the per se known methods~ sealed in it by the per ~e known method~, sterilized by gamma-radiation and stored in packaged condition at room temperature (i.e. at about.20 C)~ Under these G.ondition~, the biologically active material retained its therapeutic effiGacy during-5 years.
The data contai~ed i~ the above Examples are reported in Tables 1-3 below.

~. , ,

-4~-2f~ ~f 22~

TAaLE 1. Characteristics of a biolo~ically active material xamples Nos.
' 2 - .~nzyme content, per ccnt by weif~ht ;, 3 - textile materiaI
~1, 4 - raw materials i, j,f 5 - structure `1 6 - natural 7 - synthetiG
~' 8 - man-made ..
'`i 9 - woven 10 - non-woven 11 - knitted :
12 - im~obilized enzyme . 13 - proteolytic enzymes ' 14 - collagenolyti~ enzymes j 15 - bacteriolytiG: enzymes ;
' 16 - length of fibres, mm 17 - lint 18 - powder . ~, ' :.
.:, :

t ~

.,f . . f `f _49_ 2~ 22~

TABT,E 2. Parameters of the method for prcparing a biolo~i-.~ cally acti~e ~aterial .
1 - ~xample~ Mos..
$.;. 2 - textils material 3 - c:ellulose-containinig 4 - cellulose-non-Gontaining ?. 5 - natural .~,, 6 - man-made :
7 - syntheti~
` 8 - acti~ation ;, 9 - oxidation :
10 - hydroly~is 11 - reaction with a substance capabIe to form reacti~e ~un¢tio nal groups; K and Na periodate 12 - aldehyde reacti~e fun~ctional ~roup~;
13 - conte~t:~ reacti~e functional groupsS wt~%
14 - acid-., .
15 - washing with wate~
16 - reac.tion with a substance containing reactiYe ~unotional groups ;-, 17 - glutaric aldehyde ` 18 - dimethyl sulphatQ
. 19 - waY~ing ., 20 - washing with water 21 - washing with ethanol 22 - washing with a bu~fer solution .......

. ., -.:. .. - :

~?
.~

~50-201i22~

T.~BI~ 2 continued ... ...

1 - E~ample~ Nos.
2 - activation 3 - hydrolysis 4 - content o~ reactive functional ~roups, wt.

5 - immobilizatio~
5 - enzyme - proteolytic 8 - OEollagenolytic 9 - bacteriolytic, 10 - content, wt.~ -11 - buffer ~olution ;
12 - room temperature 13 - duration, hours 14 - covalent bond between an enzy~e and a textile ¢arrier 15 _ 13 quO e z ing- O Ut ~: ;

2~122~

TA3LE 2 continued ... ...

Examples Nos.
2 - washing 3 - drying at room temperature 4 - shaping into medicinal forms 5 - bandage~

6 - napking:

7 - lint

8 - powder g - sealing i~ a c.ontailler 10 - sterilization.
:
11 - with gamma-rays.

12 - in a gaseous ~edium ' 13 - storing~

.~ 14 - ïn paGkaged. condit:ïon ., :. 15 - at room temperature.
. , j 16 - up to 5 years.

!
.. .
.''~`` ,'.', ..
..-.
': ` '! ' ~,, . '''.

~-3 .. . .

... ~ .

.

--s2 -2 ~ 2 ~

TABL~ ~. Cllara~teri~tics of a dressing means : ,:

xampl0s Nos..
2 - protecti~e layer . 3 - textile material 4 - raw materials 5 - narural - man-made-7 - synthetic . . :
- structure

9 - wove~

10 - non-wo~en

11 - knitte~.

12 - ab-sorbing layer~

13 - medicinal form , .

14 - napkins

15 - raw materials

16 - natural

17 - man-made : :

18 - synthetic

19 - structure

20 - wo~e~

21 - non-wo~en

22 - lcnittod -~

23 - medic:inal form .~-

24 - napkins-

25 - lint:

26 - powder ::--~:
~.,. :: :: : :

,~. :

201~ 22~

T,~L~, 3 contirlued ,.. ...

1 - Examples- Nos~
2 - absorbing layer containing: an immobilized enzyme 3 - type of enzy~e~
4 - content, wt.~
5 - collagenolyti¢ enzy~nes 6 - ba~teriolytic enzyme~
7 - wound-ad~oining laye~
8 - textlle material 9 - raw materialcs 10 - natura~
11 - ma~-made 12 - synthetic 13 - structure 14 - woven 15 - non-wo~en ;~
16 - knitted 17 - medicinal form~ - napkins 18 - immobilized enzyme?
lg - proteolytic 20 - collagenolytic.
21 - bacteriolytic 22 - enzyme content, wt .~o :

Claims (25)

The embodiments of the invention in which an exclusive property or privilege is claimed, are defined as follows:

1. A biologically active material, comprising: a carrier in the form of a textile material, and an enzyme, immobilized on said carrier and covalently bound thereto, said components being taken in the following weight ratios, per cent by weight:
enzyme - 0.02 to 0.50, carrier - 99.98 to 99.50.

2. The material as claimed in Claim 1, wherein said textile material is made of natural fibres.

3. The material as claimed in Claim 1, wherein said textile material is made of synthetic fibres.

4. The material as claimed in Claim 1, wherein said textile material is made of man-made fibres.

5. The material as claimed by any of Claims. 1 through 4, wherein said textile material is selected from the groups con-sisting of a woven fabric, non-woven fabric, and a knitted fabr

6. The material as claimed by any of Claims 1 through 4, wherein, as said enzyme, use is made of a proteolytic enzyme.

7. The material as claimed by any of Claims 1 through 4, wherein, as said enzyme, use is made of a collagenolytic enzyme

8. The material as claimed by any of Claims 1 through 4, wherein, as said enzyme, use is made of a bacteriolytic enzyme.

9. The material as claimed by any of Claims 1 through 4, wherein said material is lint having fibres 1.0 to 15 mm long.

10. The material as claimed by any of Claims 1 through 4, wherein said material is a powder having a particle size of 0.001 to 1.0 mm.

11. A method for preparing a biologically active material, comprising a carrier in the form of a textile material, and an enzyme immobilized on said carrier and covalently bound thereto, said components being taken in the following weight ratios, per cent by weight: enzyme - 0.02 to 0.5; carrier -99.98 to 99.50; which method comprises the following steps of:
- activating said textile material which is chemically inert by forming therein reactive functional groups until their con-tect reaches 0.0625 to 3.125 mg-equiv. per gram of said carrier - immobilizing on said carrier thus-activated said enzyme which is a biologically active substance from a buffer solution having a pH of 6.5 to 7.5, at room temperature, for 8 to 16 hours, to form a covalent chemical bond between said carrier and said enzyme;
- squeezing-out the material thus-obtained;
- washing said squeezed-out material until no enzyme traces are detected in the washings;
- drying said washed material at room temperature (at about 20°C);
- shaping said dried material into medicinal forms;
- placing said material into a container; and - sterilizing said material.

12. The method as claimed in Claim 11, wherein, if as said textile carrier, use is made of a cellulose-containing textile material, activation of said carrier is carried out by oxidatio thereof with an alkali metal periodate to cellulose-dialdehyde containing said reactive functional groups in an amount of from 0.0625 to 3.125 mg-equiv. per gram of said carrier.

13. The method as claimed in Claim 11, wherein, if as said textile carrier, use is made of a textile material made of synthetic fibres, activation of said carrier is carried out by performing the following sequential operations of:
- hydrolyzing said carrier with an acid;
- washing said carrier thus-hydrolyzed with water until no acid traces are detected in the washings;
- processing said hydrolyzed and washed carrier with the solution of substances containing said reactive functional group until said reactive functional groups are formed within said carrier in an amount of from 0.0625 to 0.3125 mg-equiv. per gram of said carrier.

14. The method as claimed in Claim 13, wherein, as said solution of substances containing said reactive functional groups, use is made of a solution of glutaric aldehyde, and.
said carrier treated with said glutaric aldehyde solution is washed with water until no smell of said glutaric aldehyde is perceived.

15. The method as claimed in Claim 13, wherein, as said solu-tion of substances containing said reactive functional groups, use is made of a solution of dimethyl sulphate, and wherein said carrier treated with said dimethyl sulphate solution is successively washed with ethyl alcohol and with a phosphate buffer solution.

16. An individual dressing means, comprising:
- a bandage including, successively arranged therein, a pro-tective layer, an absorbing layer, and a wound-adjoining layer;
at least one of said three layers is made of a biologically active material, comprising a carrier made in the form of a textile material, and an immobilized enzyme contained in said textile material and covalently bound to said carrier, said components being present in the following weight ratios, per cent by weight: enzyme - 0.02 to 0.5; carrier - 99.98 to 99.50.

17. The individual dressing means as claimed in Claim 16, wherein said textile material is made of natural fibres.

18. The individual dressing means as claimed in Claim 16, wherein said textile material is made of synthetic fibres.

19. The individual dressing means as claimed in Claim 16, wherein said textile material is made of man-made fibres.

20. The individual dressing means as claimed by any of Claims 16 through 19, wherein said textile material is selected from the group consisting of a woven fabric, non-woven fabric, and a knitted fabric.

21. The individual dressing means as claimed by any of Claims 16 through 19, wherein, as said immobilized enzyme, use is made of a proteolytic enzyme.

22. The individual dressing means as claimed by any of Claims 16 through 19, wherein, as said enzyme, use is made of a collagenolytic enzyme.

23. The individual dressing means as claimed by any of Clai?
16 through 19, wherein, as said enzyme, use is made of a bacte?
lytic enzyme.

24. The individual dressing means as claimed in Claim 16, wherein said wound-adjoining layer is made of said biologically active material, and said absorbing layer is made of said texti?
material.

25. The individual dressing means as claimed in Claim 16, wherein said wound-adjoining layer is made of said biologically active material containing an enzyme selected from the group consisting of proteolytic enzymes, collagenolytic enzymes, and bacteriolytic enzymes, while said absorbing layer is made of a biologically active material in which use is made of an enzyme selected from the group consisting of collagenolytic and bacte-riolytic enzymes, the enzymes used in said two layers being different from one another.