Arzneimittel zur förderung der wundheilung

The available invention concerns a drug to the Iokalen application f0r the Fbrderung of the cicatrisation. It took place well-known, dal the WundheUung in several temporally following each other stages. In the stage I the Blutplasmaeiweil Fibrinogen is llt dutch Thrombin gef, kodaly it for the education of a Fibrinclots comes, which solidifies itself in presence of blood clotting factor Xlll. This first stage serves the staunching of bleeding and sealing of the Wundfl che and takes place in minutes. lm stage II cells from the Wundareal, Entzi3ndungszellen, Bindegewebszeilen and endothelial cells immigrate, to the Fibdnclot. They bidden Gef f e and as extrazellul RH matrix connective tissue, predominantly from Kollagen exist. This connective tissue called granulates ion fabrics serves as document for the education of epithelium fabric, to the Kbrperoberfl che is far it the document the epidermis. Stage II takes days until weeks and is final, if the Wundareal by Epithet, at which skin is locked by the epidermis. The cicatrisation through the stage the III, the weeks until months lasts, locked. In the course of this phase zellul ren elements increase off and the connective tissues, thus dal a firm and permanent scar fabric develop. (Bennett N.T., Schultz G.S., to. J. Surg. 1993, 165:728 - 737; Bennett N.T., Schultz G.S., to. J. Surg. 1993, 166:74 - 81). The formation of granulates ion fabric in the stage II of the Wundheilungsprozesses is caused by growth factors, which the migration and the division yon Bindegewebszellen as well as the new formation of Gef EN fbrdern and the cicatrisation to advance in this way. From the well-known growth factors in particular the Platelet is derived growth factor (PDGF), the Transforming growth factor 13 (TGF-I3), which Epidermal growth factor (EGF) and the insulin like growth factor I (IGF-I) at this Vorg ingen involved. (Bennett N.T., Schultz G.S., to. J. Surg. 1993, 165:728 - 737; Bennett N.T., Schuitz G.S., to. J. Surg. 1993, 166:74 - 81; Bhora F.Y et el., J. Surg. Res. 1995, 59:236 - 244; Lynch HE et el., Proc. Natl. Acad. Sci. The USA 1987, 84:640 - 646; Lynch HE et aE., J. Clin. Invest. 1989, 84:7696 - 7700). Also the new formation of the epidermis is caused by growth factors. They ektivieren the epidermis cells (Keratinozyten), which by the injury from the cell federation the intact basal cell layer were herausgelbst, thus dal it specific diaphragm receptors train, which adhering to the Granulationsgewebsunterlage, particularly at Fibrin Fibrinonektin ermbglichen, a provisional Ger represents {the 3st fur the migration of the Keratinozyten. (Brown G.L. etal., J. Exp. Med. 1986, 163:1319 - 1324; Brown G.L et el., N. English J. Med. 1989, 321:76 - 79). Growth factors become in the human K0rper of different fabrics and/or. Sezerniert, lm frameworks of the cicatrisation to the Thrombozyten, which store the f {3r the cicatrisation substantial growth factors PDGF, TGF {3, EGF and IGF-I in significant quantities to synthesize and in zytoplasmatischen Granula kbnnen, an important regulatorische role comes to cell kinds synthesized and into the surrounding K0rperflussigkeit. (Lynch HE et al., Proc. Natl. Acad. Sci. The USA 1987; 84:640 - 646; Ginsberg M.H et el., Thromb. Haemostas. 1988, 59:1 - 6; Hyner O.R., Thromb. Haemostas. 1991, 66:40 - 43). For the release and/or delivery of the stored growth factors from the Thrombozyten mi3ssen this by physiological stimuli, like Kollagen, Thrombin, Trypsin, ADP, serotonin or adrenalin, which eren to specific receptors to ul Oberfiache of the ThrombczytenPlasmamembran bind, being e.g. activated. The activation f0hrt to a form nderung with following aggregation of the Thrombozyten, on which these sezernieren the stored growth factors into the surrounding Kbrperfl {3ssigkeit. With most of these physiological stimuli those is the activation the following aggregation “[hrombozyten a condition far the release of the growth factors. With stimulation with Thrombin k the growth factors nnen also without Thrombozytenaggregation to be set free. (Kaplan K.L et el., Blood 1979, 53:604 - 618; Holmsen H. et al., J. Biol. Chem. 1981, 256:9393 - 9396; Philipps D.R., Baughan A.K., J. Biol. Chem. 1983, 258:10240 - 10245). To the aggregation the f (3hrenden reciprocal effects between the activated Thrombozyten and their attaching at Oberfl becomes through extrazellul RH adhesive matrix proteins chen, like e.g. Fibrinogen, Fibronectin and of Willebrand factor, mediate, which enseite to a Glykoproteinrezeptor to the Aul the plasma membrane of the activated Thrombozyten bind. A strong connection of these matrix proteins at the Receptor takes place only, if the Thrombozyten were activated above like descriptive 5s by a suitable stimulus. This complex Abl ufe of activation RK 407,484 B and aggregation of the Thrombozyten with following release of growth factors forms one of the substantial control elements in the Wundheilungsprozel (Ginsberg M.H et al., Throb. Haemostas. 1988, 59:1 - 6; Hyner O.R., Thromb. Haemostas. 1991, 66:40 - 43; Landolfi R. et al., Blood 1991, 78:377 - 381; Perschke E.I. et al., Blood 1980, 55:841 - 847; Hynes O.R., Cell 1992, 69:11 - 25; Perschke E.I., J. Lab. Clin. Med. 1994, 124:439 - 446; Savage B., Ruggeri Z.M., J. Biol. Chem. 1991, 266:11227 - 11233; Bennett J.S. et al., J. Biol. Chem. 1982, 257:8049 - 8054; Cierniewski C.S. et al., Biochim. Biophys. Acta 1982, 714:543 - 548; Philipps D.R., Baughan A.K., J. Biol. Chem. 1983, 258:10240 - 10245). Disturbances of the cicatrisation, how they vorkornrnen for instance with the sugar illness, with ven0sen or kind depressions Verschlul3krankheiten, in addition, Wundheilungsst rungen of other genesis, as for instance irradiation with radioactive substances or after burns, concern in particular the stage II of the Wundheilungsprozesses. , Dai3 in these traps the growth factors in a decreased could not be determined paints is present, sodal or groove an inferior granulates ion fabric is formed. (Dvonch V.M. et al., Surgery 1992, 112:18 - 23; Matsuoka J., Grotendorst G.R., Proc. Natl Acad. Sci. The USA 1989, 86:4416 - 4420). Umbei Wundheilungsst rungen the cicatrisation to improve, is well-known it to apply growth factors separately or in combination as pure substance or in ointment bases mixed on the Wundareal (Knighton D.R. et al., Surg. Gynecol. Obstet. 1990, 170:56 - 60; Brown G.L et al., J. Exp. Med. 1986, 163:1319 - 1324; Holmsen H. et al., J. Biol. Chem. 1981, 256:9393 - 9396). So zugef (hrten growth factors are however rapidly inactivated and/or. reduced and it enffalten its effect groove more cuber a short time interval (minutes) after the application whereby with this Pr3parationen its satisfying improvement animal cicatrisation are obtained. The WO-A - 93/25215 describe a device and a procedure for the activation of human or animal Thrombozyten and for the production of a password of Platelet factors. On filters zurDckgehaltene Thrombozyten with Thrombozyten activating L0sung contacted and (berstehende L0sung from this step, which lt also from the Thrombozyten set free Platelet factors enth, as filtrate collected, whereby zellul RH of components, as Thrombozyten or Tile of Thrombozyten, which still growth factors are contained and able, these to deliver, not into the won password of the Platelet factors (filtrate) arrive. The EP-A - a pulverous, enthSIt enriched plasma derivative describes 0,068,047 to the akzelerierten H mostase and optimized biochemical regulation of the Wundverschlusses (“fabric adhesive”) and as main components Fibrinogen, Fibrinolyse lnhibitor, Thrombin and/or Prothrombin. Zus tzlich k0nnen further additive, like Phospholipide, Prostaglandine, Trockenhalteund Stabilisierungsmittel, the however altogether no more than 1.2 Gew. - % of the Pr ready constitute, available its. AIs Phospholipid preferably client out human full blood of won Thrombozytenextrakt. In the EP-A - is among other things descriptive 0,377,855 to the Wundbehandlung the use yon from human Thrombozyten coming endothelialen growth factor (PD-ECGF), whereby alas cleaning methods for the production of the PD-ECGF several saulenchromatographische steps, einschliel lich HPLC, umfal t. Other well-known therapy to the TZE consist of covering the Wundareal with Kollagenschw mmen and/or other Pr ready ions the one st ndige humidity of the Wundareals gew hrleisten are or Reparation to use, which indegewebsschicht oberfl the chliche E the Wundareals fermentativ reduces, sodal new connective tissue from the Wundgrund to regenerate can (Nielsen P.G. et al., Acta Dermato Venerologica 1990, Suppl. 152:1 - 12; Lippert P., Wolff H., Zent.bt. Chir. 1990, 115:1175 - 1180). All this Wundverb inde and/or Pr ready ions or drugs used so far fOhren however to no satisfying success with the improvement of the cicatrisation. The available invention places itself the task to place a drug to the Verf0gung which accelerates the nat0rlichen Wundheilungsvorg nge effectively and which cicatrisation when being present Wundheilungsst0rungen, in particular also with heavy forms, compared with which so far to means and Mal3nahmen admitted improve clearly can. This task will is made available to gel0st invention in accordance with IL thereby, dal a drug to the Iokalen RK 407,484 B application f0r the F6rderung of the cicatrisation, which Thrombozyten or Tile of Thrombozyten exhibits, whereby these growth factors contained and these deliver know and in lyophilisiertem or frozen condition is present and is submitted to a procedure for the virus depletion and/or virus inactivating. In the following - if not differently indicated - the term “Thrombozyten” designates also “parts of Thrombozyten”. The invention is based to Thrombozyten, which growth factors on the realization, da6 the Iokale application yon contained and these deliver kt nnen, which can accelerate Wundheilungsvorg nge effectively. The Thrombozyten applied on the Wundareal represents a natQrliches reservoir ft to r the growth factors tigten to the F (rderung the Wundheilungsprozesse ben). To be shown, da6 the activation of the Iokal applizierten Thrombozyten has itself through in the Wundareal existing physiological stimuli with following aggregation and connection of the matrix proteins existing in the Wundareal in addition f {Jhrt, dal the growth factors stored in the Thrombozyten continuously 0ber a I ngeren period (several days) into the Wundareal be delivered. Thus obviously h0here concentrations at growth factors 0bet stands for one substantially L ngeren period as with direct gift of the growth factors in the Wundareal to the Verf0gung, whereby the Einwandem is geft rdert yon Entz ndungszellen, Bindegewebszellen and endothelial cells and the increase of these cells in the stage II of the Wundheilungsprozesses is increased. In this way it comes to a rapid and sufficient formation of granulates ion fabric, which again the training yon epithelium fabrics and the endg01tigen Wundverschlur erm (resembled. The Epithelisierungsvorgang becomes aul erdem by the set free growth factors, which the immigration and increase accelerate yon epithelium cells fSrdern, zus tzlich. Over the durability of the drug Uber a IQngeren period too gew, is preferably present the Thrombozyten in invention in accordance with l EN drug thrleisten in lyophilisiertem or tiefgefrorenero condition. For the minimization of the risk of virus infections the Thrombozyten is submitted to a procedure for the virus depletion and/or virus inactivating favourably, whereby a physical odor chemical procedure can be used odor a combination procedure. To the supply of an h (heren concentration at growth factors, in particular in the treatment of WundheilungsstSrungen, are preferential it, dal3 the content of Thrombozyten odor at parts of Thrombozyten invention in accordance with 13en medicine memo ice in such a manner is, da6 he after Rekonstitution of the Lyophiisates and/or after thawing at least 104 out, Thrombozyten per pi corresponds preferably at least. In order to obtain a particularly ausgepragte initial effect invention in accordance with IL EN of the drug immediately after application, it, in particular with heavy WundheilungsstSrungen, can be meaningful, dal the drug further growth factors exhibits, which from the Thrombozyten contained in the drug not to originate. The further growth factors k6nnen growth factors delivered stored of the same type its as yon the Thrombozyten erfindungsgemal of the EN of drug and odor a different type angeh6ren. The growth factors kSnnen with the Thrombozyten in the same Beh iltnis are present or as L0sung or Lyophilisat contained in a separate Beh Utnis its. It has itself shown, dal it in particular with ausgepr gten Filled yon Wundheilungsst0rungen is favourable, if the drug exhibits biological materials. By biological materials in the sense of the invention all materials are to be understood, which are gewebevertr iglich and absorbable and in the cooperation with the Thrombozyten or growth factors contained in the drug or unabh ngig of it the F (rderung the cicatrisation unterst0tzen. So k5nnen for example substances, which as stimuli Thrombozyten activates, and/or materials, which arrange the aggregation yon for Thrombozyten, as biological materials contained in invention in accordance with l EN drug its. In this way the effect nat {of the rlichen activating and aggregation-obtaining substances verse existing in the Wundareal is rkt, whereby the release by growth factors are erhSht and demanded the cicatrisation still more strongly. For the minimization of the risk of virus infections the biological materials are preferably submitted to a procedure for the virus depletion and/or virus inactivating, whereby a physical or chemical procedure or a combination procedure can be used. The RKs 4O7 484 B biological materials can be subjected to such procedures separately or in the mixture with other components of the Ar'zneimittels (e.g. Thrombozyten). Around the durability of the drug 0ber a I ngeren period too gew, are present the biological materials in erfindungsgemal the EN drug hrleisten favourably in lyophilisiertem or frozen s condition. The biological materials with the Thrombozyten and/or growth factors in common Beh k@nnen ltnissen are present or in separate Beh@ltnissen contained to be and the low freezing and/or Lyophilisieren of the Biomatedalien at these separately or in the mixture with other components of the drug be made. It is well-known, dab the activation and aggregation of Thrombozyten and thus the release yon in the Thrombozyten stored growth factors by the accumulation yon matrix proteins erm0glicht becomes. Outer erdem such proteins can train vemetzte structures, at which the Thrombozyten adheres and connects themselves firmly with the Wundareal, whereby these structures the diffusion of the growth factors to the Wundareal and the Einwandem of the cells from the Wundareal f rdern. Demgem@13 is characterized a preferential Ausf0hrungsform invention in accordance with EN of the drug by it, dal as biological materials fabric adhesive and/or Kollagen is intended. By fabric adhesive in the sense of the invention biological materials are understood, which consist totally or partly of interlacable proteins, which are suitable for the fabric sticking. Fibrinogen is hrend a particularly effective substance to the AusltSsung of the aggregation of activated Thrombozyten, w Thrombin one of the most effective substances for the activation of Thrombozyten represents. It is from there favourably f0r the increase of the release by growth factors and the improvement of the cicatrisation, dab the fabric adhesive out fibrinogenh ltigen proteins and Thrombin compound up is. , daP human cells, like Keratinozyten, showed up epithelium cells, embryonic and fetale cells, as well as cell components, how Liposomen, which can still accelerate demanded cicatrisation and cell zellvermehrung by Thrombozyten zus ltzlich. From there, dal3 the drug is preferential zus tzlich epithelium cells and/or Keratinozyten and/or embryonic and/or fetale cells and/or Liposomen exhibits. The cells and/or Liposomen kt nnen as fl0ssige or frozen suspension or as Lyophilisat in separate Beh ltnissen or or several of the cell kinds or Liposomen mentioned either without or with one of the other components of the drug in common Beh-*iltnissen are present. For the minimization of the risk yon virus infections the cells and/or Liposomen a procedure for the virus depletion and/or virus inactivating can be submitted, whereby a physical or chemical procedure or a combination procedure can be used. The cells and/or Liposomen can be subjected to such procedures separately or in the mixture with other components of the drug. The invention concerns also the use of Thrombozyten or parts of Thrornbozyten, which growth factors contain, for the production of a drug to the Iokalen application flat steel bar r the F rderung the cicatrisation. Preferential AusfOhrungsformen of the invention is utert in the following on the basis yon examples more nether erl. Human “rhrombozytenkonzentrat or concentrate from Thrombozytenbestandteilen is zentdfugiert by 3% sodium CIT advice anticoagulated and (1000 g/20 min), in order to enffernen plasma and other cell components. The thrombozytenreiche 0berstand and/or the 0berstand at Thrombozytenbestandteilen is suspended in RPMI medium and washed three times in RPMI medium (1000 g/20 min). The washed Thrombozyten and/or the washed Thrombozyso tenbestandteile is suspended in a RPMI medium and adjusted at least to a concentration yon 6x10s Thrombozyten and/or Thrombozytenbestandteilen per 1. The Thrombozytensuspension becomes then a virus inactivating procedure gem@l'& amp; It submitted example 3 and ends anschliel according to the procedures described below frozen or lyophilisiert whereby invention in accordance with 13es a drug will receive. Example 1: Production one invention in accordance with EN of drug RK 407,484 B Tieffrierunq: Ever 1 ml the Thrombozytensuspension is stored with -80°C within 30-40 minutes shock-frozen and frozen. Before use the Thrombozytenkonzentrat is thawed out at room temperature. Lyophilisieriung: Ever 1 ml the Thrombozytensuspension is freezingdried with -80° f r at least 24 hours frozen and anschliei3end with -20°C to -40°C {3ber 20 to 24 hours under vacuum. The freezingdried Thrombozyten is stored between -20°C and -80°C and rehydriert before use with 1 ml RPMI medium. Example 2: Production invention in accordance with r EN of a drug, the biological materials exhibits the virus-inactivated Thrombozytensuspension manufactured in accordance with example 1 a reading of interlacable human Eiweil (either Fibrinogen, Fibronectin, blood clotting factor XlII or Kollagen), which one or more procedures for the virus inactivating female example 4 have been submitted can, added, each Eiweil k0rper separately or with one another in combination, whereby the concentration of the interlacable Eiweil k (in the added L0sung preferably 70-90 mg/ml amounts to more rper soil. The Mischungsverh ltnis from Thrombozytensuspension to the Lt sung yon interlacable human Eiweil soil preferably 1:3 amount to. So erha] the tene Misohung is lyophilisiert for the achievement of a zweckmSI3igen durability in accordance with r the procedure represented in example 1 frozen or. In place of that managing described Durchft hrung the virus inactivating at the individual components to make (Thrombozyten and/or biological materials) is it also m (Jglich, the virus inactivating at the mixture of Thrombozytensuspension and Proteinl (sung in accordance with Fo the procedure of example 3. Beispie [3: Virus inactivating of the Thrombozytensuspension (photo-dynamic virus inactivating) to 50 ml in accordance with 13 the examples 1 manufactured Thrombozytensuspension is added 8-Methoxypsoralen (gelt st in Dimethylsulfoxid) up to a final concentration yon to 300 p.g/ml (final concentration of DMSO 0.3%) and illuminated with 22-27°C under a Atmosphare by 5% CO2 and 95% N2 and a pressure by 13,79 kPa (2 psi) 6 hours long with ultraviolet light from downside and above, gt sodal3 the entire Lichtintensit t 3.5 to 4.8 mW/cm2 betr (Lin L. et al., Blood 1989, 74:517 - 525). After durchgef0hrter photo inactivating in such a way received Thrombozytensuspensionen is examined for its functional Kapazit t. The functional KapazitSt becomes by measurement - Thymidinineinbaus in a FibroblastenzeUkultur determines. Example 4: Virus inactivating of the biological materials (chemical virus inactivating) biological materials, which are added in accordance with l the examples 1 manufactured Thrombozytensuspension, are virus-inactivated by the solvent Detergent method. In addition with 30°C 1% (thread/thread) trichloroethylene (n-buty] is added to a suspension of the biological materials) phosphate and 1% (thread/thread) Polyoxyethylen p isooctylphenyl ether (triton X-100) and the mixture is left 4 hours under Sch {tteln. Afterwards under additive yon 5% (VOL. /Vol.) SojabohnenSI the SolventDetergent mixture from the suspension of the biological materials at a C18-S ule (Waters milli pore) is removed by chromatography (Horowitz B. et al., Blood 1992, 79:826 - 831; Piet MP.J. et al., Transfusion 1990, 30:591 - 598; Piquet Y. et al., Vox sang. 1992, 63:251 - 256). The biological materials treated with the chemisohen virus inactivating method described above can be submitted zus tzlich still another of photo-dynamic virus inactivating in the following. Example 5: Proof animal demand of the Bindegewebsvermehrung by invention in accordance with l%e drugs 6 RKs 407,484 B the test was durchgefehrt at a fibroblast cell culture. On a cell cultural plate in accordance with f the example of 2 manufactured invention in accordance with l e drugs in a quantity was laid on yon to 200 1 per cm2 and activated dutch 50 p.I a Thrombinl0sung (3.2 IU Thrombin per ml physiological Kochselzlosung). On the laid on suspension human Fibroblesten, which originated from the 4th culture primate to 10th passage, in a density was set by 4 x 104 cells per cm2 and cultivated in a Zellku turmedium (RPMI) (culture 1). On third, and seventh day animal cultivation the ZelI Mitoserate fiJnften by measurement of the DNA synthesis 0ber the Einbeu by - thymus DIN was measured. The ZelI Mitoserate of culture 1 became with the ZelI Mitoserate another fibroblast culture (culture 2) compared, those in one RPM IN it medium, which 10 Vol% calf rum had been buried, without additive erfindungsgem& l EN of drug took place. Erqebnisse: Culture 1 showed one on the day 3 of cultivation 7fold h0heren H] - thymus DIN installation (196645 + 56864 cpm/ml) as culture 2nd on the days 5 (152749 _+ 93951 cpm/ml) and 7 (77045! 27974 cpm/ml) was - thymus DIN installation with culture 1 still 5bis 10-10-fach high as with culture 2. these differences between culture 1 and culture 2 are statistically high significant (p< 0,01) and demonstrates the F ihigkeit invention in accordance with 13en of the drug, the Bindegewebsvermehrung too fbrdern and this Aktivit tt to Dber a I ingeren period (at least 7 days) to keep upright. Example 6: Proof of the Fbrderung animal cicatrisation through alas invention in accordance with il3e drug the clinical effectiveness invention in accordance with 13en of the drug at 6 patients with chronic, do not welfare-end cutanen Ulcera of the lower Extremit iten, which had been unsuccessfully treated berets I= more lnger than a half year with surgical or conservative-local therapy, gepr ft. The Ulcera became after one of Knighton D.R. et al., Ann. Surg. 1986, 204:322 - 330, indicated Wundscore klassif iert. The Wundscore contains general parameters, anatomical conditions and Mel grbl3en of the Ulcus. The per high point number, the sch] more genuinly the conditions f0r the healing; 97 points can be achieved as maximum (=schlechteste starting situation). Behandlun.qsplan: The Ulcera was cleaned, necrotic fabric and moistened with ThrombinlOsung (3.2 IU of bovines ThrombJn/ml RPMI medium) was enffernt. Anschlie 3end was aufgefOllt the defect with in accordance with l example 2 manufactured, thawed out invention in accordance with f EN drugs and whereupon for the activation of the Thrombozyten the Thrombinlbsung in a Volumsverh, specified above, ltnis drug suspension to Thrombinl5sung by 3:1 laid on. In such a way supplied Ulcera was covered with a haffenden Wundverband (metal foil). Up to the AbheJlung the Ulcera in kind indicated above became twice per week swift ELT. The healing progress was documented photographically and histologically (Feinnadelbiopsien Jn of the 2nd and 5th treatment week). Er.qebnJsse: The patient data, urs chliche Gef f - and metabolic illnesses as well as the evaluation of the Wundscores at the beginning of treatment are in table 1 zusammengefal3t. Table 1 patient 1 2 3 4 Gef 13erkrankung metabolic sex age kindal venbs erkrankunQ Wundscore m nnlich 67 + + diabetes 51 monolith 72 + - m nnlich 69 + - diabetes 33 m nnlich 63 + - diabetes 49 RKs 407,484 B Gef IL illness metabolic patient sex age arterially vents erkrankun l Wundscore m nnlich 78 + + diabetes 63 6 like-bleached 74 - + 65a/63b A, b) two Ulcera at a leg: a) proximal, b) distal Ulcus more ber temporal operational sequence of the cicatrisation (indicated in weeks after beginning of treatment) is represented in table 2. Table 2 beginning the beginning of the Abschlul the patient Granu [ationsgewebsbildung Epithelisierun Epithelisierung 1 1st week 3rd week 8th week 2 1st week 3rd week 9th week 3 3rd week 8th week 12. Week 4 1st week 4th week 10th week 1st week no no 6 a'bl. week a6. /b3. Week a12.169. Week A, b) two Ulcera at a leg: a) proximal, b) distal Ulcus with exception of patient 3 was formed with all patients of the Ulcusboden hsr already in the first treatment week a well supplied with blood granulates ion fabric, DOS after further treatments with invention in accordance with 13en the drug until approximately two weeks after beginning of therapy increased and DOS Ulcus ausf llte. , Dal with all patients naeh the first treatments the environment of the Ulcus was remarkable already calmed down, to DOS erythema and the Odem of the surrounding skin disappeared and ouch the Ulcusrand any longer dematSs and me colors was not. Histologically zellreiehes showed up, predominantly from fibroblasts and Fibrozyten existing granulates ion fabric with reichlJcher Gef il3neubildung and kollagener fiber formation and with only oberfl chlich small infiltration of Entz0ndungszellen and tissue necroses with all Biopsien in the second treatment week. A Epithelisierung of the skin defects proceeded after the third treatment week from the Wundr ndern and could then ouch histologically with the second Biopsien in the f (nften treatment week to be proven. In the further process of the treatment took DOS paint the Ulcera on the one hand dureh the Epithelisierung, in addition, by a scarred contraction off. They healed 5 with exception with patient spatestens in the 12. Treatment week scarred off. Those angefL hrten above results show, dal the Iokale application invention in accordance with l EN of drug with patients, who had been unsuccessfully treated with conservative therapy at least a half year long and from there extremely bad conditions far a cicatrisation exhibit, the cicatrisation demand and in dJese way chronically not-welfare-end, cutane Ulcera to healing completely to bring can. 1,