PROCEDURES FOR MANUFACTURING (AMIDO) NSUBSTITUIERTEN BLEOMYCINEN

The invention refers to a procedure for manufacturing (amido) N-substituted Bleomycinen of the general formula CONH2] H2 CH2 .NH CH CH CH2/CO-NH-X N-- IL=J n ' --H CH CH2 CO-NH (CH 2) 3-N (CH2) 3-N-R 2 (l) where CBM a remainder of the Bleomycingerüstes represents, X a) an alkyl group or a b) a Aminoalkylgruppe meant, where the amino group with or two alkyl groups with 1 to 5 carbon atoms is substituted and the alkyl groups at the amino group exhibit a not substituted Niedrigalkylamino, Phenylniedrigalkylaminooder Halogenphenylniedrigalkylaminosubstituenten k6nt0 nen, [: [1 for 1. a Cycloalkylgruppe, 2. one by an alkyl group replaced if necessary with at least a halogen substituted Phenylgruppe or 3. one by at least one of the substituents halogen, Cyano and/or t5 Benzyloxy substituted Phenylgruppe stands if necessary and for R2 a hydrogen atom or a group of the formula - CH2 - g 1 meant. This (amido) N-substituted Bleomycine is characterised particularly by its low toxicity. Bleomycin is antibiotic substances, those in the year 1966 of Umezawa, carcinostatisch working a family by, one the inventor of the article of the patent, and its coworkers was found (Umezawa et al., journal OF Antibiotics, 19A, S.200, 1966). Bleomycin is produced by Streptomyces verticillus, a Actinomycete, and is a basic, water-soluble Glycopeptid, which readily for each molecule an atom of bivalent copper chelatisiert. With usual cultivating 16 members of the Bleomycinfamilie develop, from which everyone was isolated Is. for example Umezawa et al., journal OF Antibiotics, 19A, S.210 (1966)]. By this Bleomycinen the Bleomycine A1, a2, AS, B 2 and Demethyl-A2 becomes applied in form of a copper-free mixture (which is called in the following Bleomycinkomplex) clinically with the cancer therapy and with special success with the treatment by flaky (squamösem) Zellcarcinom, in addition, with the treatment by skin cancer, head cancer, neck cancer, cancer of the lungs and malicious lymphoma at present. Different Bleomycine is also in the US-PS Nr.3, 922.262 and in the US-PS RH 30.451 beschrisbon. With the usual Fermentieren this Bleomycine usually develops in kupfsrhaltiger form. The copper-free form will receive from the cupreous form by removing the copper. Soferne nothing else is indicated, in the following the expression “Bleomycin” both for the cupreous form and for the copper-free form is used. The Bleomycine specified above corresponds to the general formula I 0 H OH I HO 2OH J H C NH2 (II) in which R a finalconstant amine remainder of the Bleomycins one meant and one omitted chelatisierte copper for the case of the cupreous form. The above Bleomycine possesses excellent effectiveness, in addition, depending upon administration way Nehenwirkungen such as lung toxicity, why their application is reduced at present. It was desired to develop Bleomycine with smaller side effects. In the effort to correspond to this need in the course of expanded investigations new (amido) N-substituted Bleomycine was found, which the above general formula (I) correspond. From the BE-PS Nr.895498 such (amido) N-substituted Bleomycine is become known, however the procedures for their production, described there, are multistage and from there complex. A goal of the invention was it from there to create a procedure for the production of such (amido) N-substituted Bleomycine which proceed from a Bleomycinderivat simply accessible by fermentation. To the solution this: Task consists the invention of that (amido) a N-substituted derivative 8 - [N-methyl-n (3-aminopropyl) - amino l-propylaminobleomycins the general formula of the iONH2 iH2 CH2 CH /NH. CH2 /CH " CO-NH-X CBM N =, m_l I I! IL H -- CO-NH (OH 2) is condensed 3-N (CH2) 3 - NH 2 (nl) where CBM possesses the meaning indicated above and an alkyl group substituted if necessary represents to X, with an aldehyde of the general formula RICHO, (IV) where RI possesses the meaning indicated above, under reduction with a boron hydride connection, until (triphenylphosphin) - tetrahydrohorat copper or palladium/activated charcoal. In the general formula (1) appearing remainder of CBM of the Bleomycingerüstes the lines framed part ierten in the general formula (IL) the Bleomycine with Strich1 and both the cupreous form and the copper-free form are covered. I0 to applicable the according to invention aldehydes of the general formula (IV) count for example Cycloalkylaldehyde such as Cycloalkylaldehyde with 5 to 13 carbon atoms and in particular Cyclopentylaldehyd, Cyclooctylaldehyd, Cyclododecanoylaldehyd, Cycloundecanoylaldehyd etc., by a Phenylgruppe of substituted satisfied aliphatic aldehydes, whose Phenylsubstituent can be substituted, as for example a Phenylgruppe as substituent exhibiting satisfied aliphatic aldehydes such as Phenylacetaldehyd, Phenyläthylaldehyd and halogeneous-substituted Phenylacetaldehyde, for example Chlorphenylacetaldehyd, and Benzaldehyde, for example Benzaldehyd, Halogenbenzaldehyde (e.g. p-Chlorbenzaldehyd or m-Brombenzaldehyd), Cyanobenzaldehyd and Benzyloxybenzaldehyde (e.g. m, p-Dibenzyloxybenzaldehyd). The one which can be used the connection of of the general formula (III) it can be manufactured by the fact that 3 [N-methyl-n (3-aminopropyl) - amino], in accordance with the invention as basic material - propylaminobleomycin (which is called in the following Bleomycin AP P) with a Bleomycin inactivating enzyme (which is called in the following inactivating enzyme, Umezawa et al., journal OF Antibiotics, Bd.27, S.419, 1974) to the 3 [N-methyl-n (3-aminopropyl) - amino] - propylaminodeamidobleomycin (which is called in the following Deamidobleomycin APMP) the formula iONH2 iH2 CH2 NH CH/CH2/“COOH one hydrolyzes and then this Deamidobleomycin APMP with an amine the general formula NH2 - X, (V) where X for the general formula (I) indicated meaning possesses, one condenses. When examples of the remainder of X know alkyl groups with 6 to 10 carbon atoms and a primary amino group as substituent exhibiting alkyl groups with 2 to 6 carbon atoms are called, whereby the n-Octyl~, the Phenyläthylaminopropyl, which 3 [[N-methyl-n {3 [2 (p - chlorphenyl) - ethyl] - aminopropyl} - amino]] - propylund the Di-n-butylaminopropylgruppe to be particularly preferred. The connection of the general formula (III) can be manufactured for example as follows. To receive cattle liver homogenized with a phosphate buffer solution, on which the received Hcmoge -1 l0 is centrifuged nisat at a number of revolutions of 8000 min and the supernatant liquid is dialysiert against a phosphate buffer solution, in order a raw enzyme solution. The copper-free form of the Bleomycin APMP solved in a phosphate buffer solution is then added to this raw enzyme solution, on which the received mixture during 5 to 48 h with 37°C converted and the received reaction mixture to a suitable method, for example by adding tri chlorine acetic acid (in the following called TCA) up to obtaining a concentration of 5 Gew. - % and Abzentrifugieren developed striking down, from proteins is released, whereby the precipitation 3mal with 5% TCA is washed and the supernatant liquid is combined with all washing liquids. After neutralizing copper acetate in surplus referred to Bleomycin is added to the reaction mixture released from proteins, with which the desired connection is transferred in a Kupferchelat. Salts are separated thereby that the solution of the Kupferchelats into one with an adsorption resin, u.zw. Diaion (AP-40 as well as distilled water, filled column and thus the desired connection at the resin is poured is adsorbed and afterwards the salts from the column with distilled water to be out-washed. The column becomes then with one in the VOL around relationship of 1: 4 from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture eluiert, whereby the parliamentary group showing an absorption maximum with a wavelength of approximately 290 Nm are caught. Combined with one another the parliamentary groups become after neutralizing with an anion exchange resin, u.zw. Dowexq D 44 (OH-type, manufactured of Dow Chemical Co.), neutralized, then under reduced pressure restricted and finally freezingdried. The received powder solved in distilled water, on which the received solution into one with Cm-Sephadex () C-25 (Na+ type to cause manufactured of Pharmacia Fine chemicals CO.), which with a buffer solution possessing a pH value of 4,5 from 1/20 m-acetic acid and Natriumacetat in the equilibrium had before been brought, filled column one pours, in order adsorption. The column is then eluiert a Gradientenelution with linear sodium chloride concentration rising to 1,0molar by adding from sodium chloride to mentioned above the Pufferl6sung, whereby the desired connection containing bluish-violet colored parliamentary groups are caught. Combined with one another the parliamentary groups are entsalzt according to the method described before using Diaion () HP-40 and freezingdried then, with which the cupreous form will receive from Deamidobleomycin APMP as blue amorphous powder. In such a way received Deamidobleomycin APMP becomes then with an amine of the general formula NH2-X, (V) in actually well-known way, for example after in the US-PS Nr.4, 195.018, to an acid amide condensed, which is released if required from copper, with which the connection of the general formula (III) containing reaction mixture is received. The above condensation reaction becomes using a condensation means such as Cyclohexylcarbodiimid, 1-Äthyl-3 (3-dimethylaminopropyl) - carbodiimid, 1-Cyclohexyl-3 (2-morpholinoäthyl) - carbodiimid, Diisopropylcarbodiimid, Diphenylphosphorazidat (DPPA), Diäthylphosphoroeyanidat (DEPC), 6-Chlor-la (p-chlorine-benzosulfonyloxy) - benzotriazol (CCBT) cd.dgl, accomplished. Together with the above condensation means a condensation auxiliary material can be used such as p-NitropheSO nol, o, p-Dinitrophenol, Pentachlorphenol, 2,4,5-Trichlorphenol, Pentafluorphenol, N-Hydroxysuccinimid, N-Hydroxy-5-norbornen-2,3-dicarboximid, 1-Hydroxybenzotriazol cd.dgl. The condensation in a solvent such as water, dimethylformamide, acetonitrile or a mixture hievon at a reaction temperature from -20 to +50°C, preferably -10 to +30°C, accomplished, whereby the conversion duration is subject actually to no special restriction, however one conversion duration from i to 70 h sufficiently is. The connection of the general formula (III) from the reaction mixture in the following way one isolates. From the reaction mixture the desired connection is precipitated by adding an organic solvent such as acetone, ether cd.dgl, on which the precipitation solved in distilled water, adjusted to a pH value of 6 and then into one out-washed with distilled water and an adsorption resin poured the received solution such as amberlites () XAD-2 (manufactured pipes le & Haas CO.) filled column, from the column the salts with distilled water and the desired connection with sour, adsorbed at the adsorption resin, aqueous methanol, for example with one in the VOL around relationship 1: 4 from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture, is eluiert, whereby an absorption maximum with a wavelength by approximately 290 Nm parliamentary groups showing are caught. Combined with one another the parliamentary groups are neutralized, restricted and freezingdried with an anion exchange resin, e.g. Dowex () 44 (OH-type, manufactured of Dow Chemical Co.), with which the connection of the general formula (III) in raw form and in form of a powder is received. It can be the case that the Entsalzen described above is dispensable. In this case the precipitation specified above is loosened in distilled water and then in the following way continues working. The powder specified above is loosened in distilled water, on which the received solution into one with CNI Sephadex) C-25 (Na+ type, manufactured of Pharmacia Fine chemicals CO.), which with a pH value vcon 4.5 possessing buffer solution on 1/20 m-acetic acid and Natriumacetat in the equilibrium had before been brought, filled column one pours and the column according to a linear density gradient method one eluiert, as continuously sodium chloride up to the rise of the sodium chloride concentration to 1,0incler is added to the buffer solution specified above. Since not converted materials and by-products are already in former times eluiert, these materials can be separated using an ultraviolet absorption monitor. If the desired connection containing parliamentary groups contain still of impurities, this 3e is completely separated by repeating the above Chromatographierschrittes. The above Chromatographieren can be replaced by a Chromatographieren using an adsorption resin such as amberlites () XAD-2. With this Chromatographieren an aqueous solution of the raw product is poured into a column, which is filled with a resin for example blended with teiger aqueous Ammonacetatlösung, in order to adsorb the desired connection at the resin. The Eluieren of the column takes place according to a gradient method with linear rising concentration, whereby continuously under constant increase of the methanol concentration methanol is added to the above buffer solution. Since in this case not converted materials are eluiert later in former times and by-products, these materials can be separated and removed using an ultraviolet absorption monitor. If it turns out that the desired 4e connection contains still impurities is repeated, the above Chromatographieren for complete separating of the impurities. The two cleaning methods described above can be used separately or in combination with one another. In such a way received parliamentary group of the desired connection becomes in the way described above using an adsorption resin such as amberlites (XAD-2 it entsalzt and then freezingdried with which the cupreous form of the connection of the general formula (III) as blue amorphous powder is received. This cupreous form can according to according to a well-known method, for example in the US-PS Nr.3, 929.995 described and method accomplished using EDTA, 5Q into the copper-free form be transferred. An example of the transformation into the copper [reie form in the following one describes. The cupreous form is solved in distilled water, on which the received solution is poured into a I} 4) with amberlites 'XAD-2 and with distilled water filled column, in order to cause adsorption. The resin in the column is then washed with 5% of the DIN atrium salt an aqueous sodium chlorid solution containing of Äthylendiamintetraessigsäure (in the following called EDTA.2 well), whereby Kupferion is exhausted by EDTA.2 well and the copper-free form at the resin remains. From the column EDTA.2 will then wash well with sodium chloride herausgeS, on which the column with distilled water is flushed. Then becomes the column with sourer, aqueous methanol, for example with one in the VOL around relationship of i: 4 from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture, eluiert, whereby a Absorptionsmaxiraum with a wavelength by approximately 290 nmzeigende parliamentary groups are caught. The pH value combined with one another of the parliamentary groups is then stopped by means of Dowex () 44 (OH-type, manufactured of I0 Dow Chemieal CO.) to 6,0, on which the parliamentary groups under reduced pressure are restricted and freezingdried, with which a hydrochloride of the copper-free form of the connection of the general formula (III) as white amorphous powder is received. If with the function described above instead of aqueous hydrochloric acid aqueous sulfuric acid is used, a sulfate is received. By appropriate choice of the acid with the above Eluierschritt thus a salt with any desired acid can be received. When working in the procedure according to invention in such a way received connection of the general formula (llI) with an aldehyde of the general formula (IV) is condensed reducing. As reducing agent usable for this condensation can boron hydride connections, e.g. Natriumcyanoborhydrid, sodium boron hydride, lithium boron hydride or potassium boron hydride, until (triphenylphosphin) is called - tetrahydroboratokupfer etc. A catalytic reduction can be made by means of a catalyst such as palladium on carbon. The aldehyde of the general formula (IV) supplies i up to 1.5 mol, per mol of the connection of the general formula (III) connections of the general formula (1) with use in a quantity of 2 mol or less, preferably where R2 essentially hydrogen meant. When using 3 mol or more of the aldehyde a connection of the general formula (1) develops, where R2 stands for - CH2 - g 1. Those desirable-proves quantity of aldehyde, which can be used, amounts to 1.2 mol in case of the production of a connection of the general formula (1) with the meaning from hydrogen for R2 and 5 to mol, preferably about 10 mol, for the case of the production of a connection of the general formula (I) with the meaning of - CH2 - R [for R2. If the aldehyde of the general formula (IV) is with difficulty soluble so as for example m, p-Dibenzyloxy-benzaldehyd in methanol, the quantity of this aldehyde under the condition can be reduced that the conversion duration is extended. Soferne the solvent at the conversion does not participate, exists no special restriction regarding the solvent used with the above condensation reaction. As solvents polar solvents are used such as water, methanol, ethanol, dimethylformamide, Dimethylsulfcxyd, tetrahydrofurane and acetonitrile and mixtures hievon etc. The conversion temperature usually amounts to -5 to +80°C and 0 to +50°C. It is in particular desired, on production of a connection of the general formula to work (1) with the meaning of hydrogen for R2 between 0 and 25°C and on production of a connection of the general formula (1) with the meaning of - CH2 - g 1 for R2 between 35 and 50°C and this particularly on use of a strongly sterisch prevented or weakly soluble aldehyde. The conversion duration lies be suitable-proves between 3 and 70 h. In the case of use of a strongly sterisch prevented or only weakly soluble aldehyde a longer conversion duration of for example 150 h than zweekentsprechend proves. From in such a way received reaction mixture the received connection of the general formula (1) in the following way becomes isolated. If a boron hydride connection were used, the reaction mixture is stopped for decomposition of the reducing agent with hydrochloric acid, available in the surplus, to a pH value of i and at ambient temperature 5 to 10 min agitated, then neutralized, by distilling under reduced pressure from methanol liberating, by extracting with ether or butanol from überschüssigern aldehyde and Keton liberating and afterwards entsalzt, by in such a way prepared ReaktionsgeI} 4} mix into one with an adsorption resin such as amberlites XAD-2 (manufactured from Rohm &} {carrion CO.) and with distilled water filled column is poured and then salt from the desired connection adsorbs containing resin containing column with distilled water is out-washed. The Eluieren becomes then methanol aqueous with sourer, for example one in the VOL around relationship of 1: 4 from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture, made, whereby a blue Bleomycinderivat containing parliamentary groups are caught. Combined with one another the parliamentary groups are neutralized if required with an anion exchanger resin such as Dowex () 44 (OH-type, manufactured of Dow Chemical Co.), restricted and freezingdried under reduced pressure, whereby the desired connection in raw form and as blue powder will receive. The further cleaning of this raw connection is made as follows. lB in the above way the received powder is poured solved in distilled water, on which the received solution had been brought to 5 possessing buffer solution before into one with Cm-Sephadex () C-25 (Na+ type, manufactured of Pharmacia Fine chemicals CO.), soft with a pH value of 4=, from 1/20 n-acetic acid and Natriumacetat in the equilibrium, filled column, in order to cause adsorption. The column is then eluiert according to a gradient method with linear rising sodium chloride concentration, whereby constantly sodium chloride is added to the above buffer solution, until the sodium chloride concentration constantly rose to 1,0molar. If the desired connection containing parliamentary groups contain still of impurities, still another Chromatographieren knows using an adsorption resin such as AmberliteL) XAD-2 to follow on the above Chromato2 raphieren. Combined with one another the parliamentary groups are poured, for example together with an aqueous 4%igen Ammonaeetatlösung, into a column filled with a resin, in order to adsorb the desired connection at the resin. The column is then eluiert according to a gradient method with linear rising methanol concentration, whereby for constant increase of the methanol concentration constantly methanol is added to the above buffer solution. In this case not converted materials first it eluiert on which the connection with the meaning of hydrogen for R2 is also eluiert last and then the connection the same meaning for RI and R2. The separation of these parliamentary groups takes place using an ultraviolet monitor in satisfactory way. If the desired connection containing parliamentary groups contain still of impurities, for complete separating of the impurities the above Chromatographieren is repeated. During this processing step can be replaced the column filled with Cm-Sephadex () C-25 by one with Cm-Sepharose () to CL-6B (manufactured of Pharmacia Fine chemicals CO.) or Cm-Toyopearl () 650 (manufactured of Toyo soda MFG CO.) od.dgl, filled column, whereby these resins are washed with methanol. In this case the demineralization stage can be void after the conversion. The reaction mixture is poured directly into a column, in order to adsorb the desired connection. After the reagents and other materials from the column were out-washed with methanol, the column becomes also at Natriumch! orid 0,1molarem aqueous methanol eluiert. Combined with one another, the desired connection containing parliamentary groups are chromatographiert to amberlites () XAD-2. The desired connection containing and united with one another parliamentary groups in the way described above using halls () HP-40 are entsalzt and freezingdried then, with which (amido) Bleomyeins of the general formula (I) N-substituted the cupreous form when blue amorphous powder will receive. In such a way received cupreous form (amido) of a N-substituted Bleomycins can be released according to the method described above from copper, in order to receive a desired salt of a copper-free form as white amorphous powder. Typical examples of producible according to invention connections are mentioned in table I, whereby the physicochemical characteristics of these connections in table II appear. Table I connection name of the connection abbreviation No. 1 ll 12 13 3 {N-methyl-n [3 (p-chlorbenzylamino) - propyl] - - amino} - propylamino (amido) n {3 - [(s) of -1 “- pheny! - äthylamino] - propyl} - BLM 3 {N-Methy! - N [3 (p-chlorbenzylamino) - propy! ] - amino} - propylamino (amido) n [octy! ] - BLM 3 [[N-methyl-n {3 - [2 (p-chlorpbenyl) - äthylamino] - propyI} - amino]] - propylamino (amido) n - {3 - [(s) of -1” - phenyläthylamino] - per pyl} - BLM 3 [[N-Methy! - N {3 - [2 (p-CH! orphenyl) - äthylamino] - propyl} - amino]] - propylamino - (amido) n - BLM 3 {N-Methy! - N [3 - (p-cyanobenzylmethylamino) - - propyl] - amino} - propylamino (amido) n - {3 - [(s) of -1 “- phenyläthylamino] - propy! } - BLM 3 {N-methyl-n [3 (p-cyanobenzylmethylamino) - - propyl] - amino} - propylamino (amido) n - - BLM 3 {N-methyl-n [3 - (cyclooctylmethylamlno) - - propyl] - amino} - propylamino (amido) n - {3 [(S) - 1” - phenyläthy! ] - aminopropy! } - BLM 3 {N-methyl-n [3 (cyclooctylmethylamino) - - propyl] - amino} - propylamino (amido) n - [octy! ] - BLM 3 - {N-methyl-n [3-bis (m, p-dibenzyloxybenzylamino) - propyl] - amino} - propylamino (amido) n - {3 - [(S) - I '- phenyläthylamino] - propyl} - BLM 3 - {N-methyl-n [3-bis (m, p-dibenzyloxybenzylamino) - propy! ] - amino} - propylamino (amido) n - [n-octyl] - BLM 3 [[N - methyl n - {3 - [2 - (p - chlorphenyl) - äthy! - amino] - propyl} - amino]] - propylamino (amido) n -4 {3 [[N - methyl - n {3 - [2 - (p - chlorphenyl) - äthylamino] - propyl} - amino]] - propyl} - BLM 3 [[N-methyl-n {3 - [2 (p-chlorphenyl) - äthylamino] - propyl} - amino]] - propylamino (amido) n - [3-Adi-n-butylamino) - propy! ] - BLM 3 - {N-methyl-n [3 (p-cyanobenzylamino) - propyl] - amino} - propylamino (amido) n [3 (DIN - butylamino) - propyl] - BLM Nr.380886 dMCLBZ PEP dMCLBZ OCT dMCLPE PEP dMCLPE OCT dMCNBZ PEP dMCNBZ OCT dMCO PEP dMCO OCT dDD PEP dDD OCT dMCLPE MCLPE dMCLPE EPA dMCNBZ BPA] abelie [[connection No. 1 2 3 4 6 7 8 9 I0 11 12 13 abbreviation dMCLBZ PEP dMCLBZ OCT dMCLPE PEP dMCLPE OCT dMCNBZ PEP dMCNBZ OCT dMCO PEP dMCO OCl dOD PEP dOO OCT dMCLPE MCLPE dMCLPE BPA dMCN8Z-8PA UV absorption maximum that form n (E i%/I cm) 289 (go) 290 (88) 290 (7o) 290 (85) 29l (93) cu-fre£en 292 (94) 291 (EEC) 291 (83) 285 (74) 285 (7e) 290 (79) 29o (83) 290 (81) Oünnschzchtchromatographle t of the cu-containing form Rf 0.53 0.50 0.53 0.52 0.69 0.59 0.53 0.42 0.09 0.04 0.25 0.60 0.70 electrophoresis +2 cu-containing forums, Rm (Rm of alanine = 1,0) 1,16 1.08 1.04 0.96 1.10 0.95 1 WHETHER 0.96 0.93 0.64 1.20 1.11 1.10 Anmerkun9: +1 Silicagel 60 F 254 silanisiertQ (Merck CO.) Methanol-6%ige Ammoniumacetatlö$ung (60: 40) (V/V) +2 Avlce! SFQ (FMC CO.), AmelsensSure acetic acid water (27: 75: gO0 V/V), 800 V, 15 min in the following are described the biological characteristics of the typical, producible according to invention connections. 1. Antimicrobial effect against Mycobacterium smegmatis ATCC 607 and Bacillus subtilis the antimicrobial effect against the test organisms mentioned was determined according to the agar plate cylinder method. The effect was indicated on the assumption that dle effect of a standard sample of Bleomycin a2 (copper-free form) amounts to I000 g Wirkungsstärke/mg. 2. Growth inhibition on a culture of (Hela S3) - cells a medium in a Petri plate made of plastic (MEM with 10% calf serum) became with (Hela S3) - cells inoculates. Two days after into the Petri plate a Bleomycin was brought. After 3täßigem cultivating the cells were counted out. The Wachstumsinhibiti0n was computed after the following equation Wachstumsinhibitlon (%) = 100 (B-A)/(B-C) in per cent, in which The schließliche number of the cells meant three days after adding the test connection, B the schließliche number of the cells with without test connection the accomplished comparison attempt and C the number of cells at the time of adding the test connection. ID50 - ii - was received Nr.380886 by laying the concentration of the test connection on against growth inhibition expressed in per cent. With the examinations in accordance with the above points the 1 and 2 received results is shown in table III. Table III 1 2 3 4 6 7 8 g 11 12 13 abbreviation antimlkroßielle effectiveness of of the cu-free connection (pg W£rksamkeit/mg) approximately against Mycobacterium smegmatZs Baclllus subtZlis ATCC 6O7 [nhibitionskonzentration that cu-frelen connection for 50%ige growth prevention of kultiv£erteù (HeLa S) - cells dMCLBZ PEP dMCLPE OCT dMCLP£ PEP dMCLP£ OCT dMCNBZ PEP dMCNBZ OCT dMCO PEP dMCO OCT dOD PEP dOD OCT dMCLP£ CLPE dMCLPE BPA dMCN8Z-BPA 14880 38200 15593 38750 21877 49270 54345 65733 115,153 65O 31690 14730 2500 1808 4267 1898 11056 381 7273 5473 23 13 270 36g0 4755 pg/ml 0.53 0.43 0.37 0.23 0.60 0.28 0.33 0.23 0.058 0.30 0.30 0.40 1.10 3rd lung toxicity (Pulmonarfibrose) in mice it were used 15 weeks old male mice by the ICR trunk, those in groups were combined into ever 9 mice. Each test preparation was intraperitoneal injected during 10 following each other days lmal daily in a dose of 5 mg/kg. After final administration the mice were nourished 5 weeks under observation, slaughtered then and subjected to a Autopsie, in order to determine the occurrence and the extent of the Pulmonarfibrose. The evaluation took place via comparison of the number at Pulmonarfibrose got sick treated mice and the weight of the illness. The received results are shown in table IV. The weight of the illness was evaluated in terms of figures to points in the following way. Number of points 0: no Fibrose 1: Enrichment of Exudat in the alveoles and fibroseartige changes in the Alveolarseptum 2nd Fibrose in several ranges 4: scattered Fibrose 6th Fibrose in more than 2/3 of the entire range the “relationship” appearing in table IV was computed by comparison with the “Bleomycinkomplex”. Table IV connection No. '1 3 6 7 8 g! 0 illness number of mice with Pulmonarfibrose 21g (20) 0/1 (0) o19 (o) Olg (o) olg (o) Olg (o) olg (o) extent total number at PulmonarFibrose/Gesamtzah! at samples 2127 (7) 013 (0) 0127 (0) 0127 (0) 0127 (0) 0127 (0) 0127 (0) relationship 0.07 0 0 0 0 0 0 like the above results, possess the producible according to invention show connections a strong waehstumshemmende effect opposite cultivated (Hela-S3) - cells and an expressed antimicrobial effect with extremely small Pulmonartoxizität, with which these connections for their clinical use are recommended. The invention is in detail described in the following by the invention in no way restrictive examples. Examples: A gradates 800 g of fresh cattle liver became with 1,6 1 a pH value of 7,2 possessing 0,05m - l phosphate buffer solution homogenizes, on which the Nomogenisat was centrifuged at a number of revolutions of 8000 min min and the supernatant liquid was dialysiert using a cellulose diaphragm against 0,05m-Phosphatpufferlösung, with which a raw enzyme solution became to receive. Then 32.7 g B became [eomycin APMP with 1,5 1 of the raw enzyme solution shifted, on which the received mixture was converted with 37°C during 24 h and shifted then for precipitating proteins with 150 ml 55%iger tri chlorine acetic acid (in the following called TCA). The developed precipitation was abzentrifugiert and washed 3mal with 5% TCA. The supernatant Flüsz0 sigkeit and all washing liquids was combined with one another, on which the whole with 4m-NaOH neutralized and the neutralized solution with 5 g copper acetate (1.2 equivalents related to Bleomycin) was shifted, with which the desired connection was transferred in a Kupferchelat. For Entsalzens of the Kupferchelats the solution of the same became into one with the adsorption resin Diaion (HP-40 (manufactured of Mitsubishi Chemical CO.) and water filled column of 7 1 contents poured, in order to adsorb the desired connection at the resin. The salt out-washed from the column with 14 1 distilled water. The column became then with one in the VOL around relationship of 1: 4 was neutralized from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture eluiert, whereby the one maximum adsorption combines with a wavelength of approximately 290 nmzeigenden parliamentary groups caught, with one another, with Dowex () 44 (OH-type, manufactured of Dow Chemical Co.) and restricted then under reduced pressure. The received concentrate became into one with Cm-Sephadex C-25 (Na+ type, manufactured of Pharmacia Fine chemicals CO.), which with a buffer solution possessing a pH value of 4,5 from 1/20 m-acetic acid and Natriumacetat in the equilibrium had before been brought, filled column of 3! Contents poured, in order to cause adsorption. The column was then eluiert according to a gradient method with linear rising sodium chloride concentration, whereby constantly sodium chloride was added to the above buffer solution, until the sodium concentration had constantly risen to 1,0molar. The blue parliamentary groups caught with a sodium concentration of for instance 0,6molar were combined with one another and entsalzt and freezingdried according to the method described before using Diaion () HP-40, with which 21.7 g of the cupreous form became to receive from Deamidobleomycin APMP as blue amorphous powder. The yield amounted to 66%. This blue powder showed in distilled water ultraviolet absorption maxima (E 1%/1 cm) with 241 Nm (127) and 292 Nm (104). This powder showed when measurement in the KBr tablet infrared absorption maxima (as Wellenzahl, cm-1) with 3350, 1725, 1650, 1560, 1460, 1380, 1200, 1140, 1100, 1060, 1020, 990, 880 and 770. Further physically chemical characteristics are indicated in table V. B gradates is in i00 ml dimethylformamide became 5 g of the cupreous form of Deamidob, received in accordance with stage A] eomycin APMP and 8.3 g (10 equivalents per mol of Bleomycin) 3 [(S) - l' Phenyläthylamino] - propylamin. Hydrochloride (in the following called PEP Amin.Hydrochlorid) solved, on which the received solution was gekehlt to 0°C and then with 13,4 g (30 equivalents per mol of Bleomycin) 1-Hydroxy-benzotriazol and 3.63 ml N-methyl-morpholin and then still with 6,8 g (10 equivalents per mol of Bleomycin) Dicyclohexylcarbodiimid shifted and the mixture under agitating at ambient temperature during 6 h was converted. By shifting the reaction mixture with the 10lachen of its volume to acetone the desired connection was precipitated, on which the received precipitation was washed thoroughly with acetone and solved in distilled water. The received solution poured into one with Cm-Sephadex C-25 (type of well, manufactured of Pharmacia Fine chemicals CO.), which with a buffer solution from 1/20, possessing a pH value of 4,5, m-acetic acid and Natriumacetat in the equilibrium one had before brought, filled column of 500 ml contents, in order to cause adsorption. The column was then eluiert according to a gradient method with linear rising sodium chloride concentration, whereby constantly sodium chloride was added to the buffer solution specified above, until the sodium concentration had constantly risen to 1,0molar. The parliamentary groups eluierten with a sodium concentration of for instance 0,7molar were caught and compatible with one another (colored blue) and according to the method using Diaion, described before, “HP-40 (assigned quantity of 500 valley) entsalzt and then freezingdried, with which 3.3 g of of the cupreous form of 3 (N-Methylaminopropyl) - aminopropylamino (amides) n {3 - [(S) - l' phenyläthylamino] - propyl} - bleomycin (in the following called dAPMP PEP) as blue amorphous powder and with a yield from 66% was received. This connection showed when measurement in distilled water ultraviolet absorption maxima (E i /.i cm) with 241 Nm (108) and 292 Nm (87). This - i connection showed measured infrared absorption maxima (Wellenzahl, cm) in the KBr tablet with 3360, 1725, 1645, 1560, 1460, 1380, 1250, 1140, 1100, 1060, 1015, 990, 880, 765 and 700. Further physicochemical characteristics are shown in table V. If in stage B of the above function PEP amin.Hydrochlorid and N-methyl-morpholin by n-Oatylamin one replaced, that became 3 (N-Methylaminopropy!)- aminopropyIamino (amido) n (n-octyl) - receive bleomycin (in the following called dAPMP OCT). This connection showed measured ultraviolet absorption maxima (E 1%/1 cm) in distilled water with 241 Nm (118) and 292 Nm (95). This connection showed measured infrared absorption maxima (waving -1 number, cm) in the KBr tablet with 3350, 2940, 1720, 1640, 1555, 1460, 1370, 1250, 1115, 1100, 1060, 1010, 995, 930, 880 and 760. Further physicochemical characteristics are shown in table V. Table V synthesized Oünnschichtchromatographie +I electrophoresis that cu-haltig n connection of the cu-haltigen form form, Rm R (Rm of alanine = 1.0) Oeamldobleomycin APMP dAPMP PEP dAPMP GCT 0.28 0.37 B, 93 1.17 1.00 +1 $11icagel GO F 254 silanisiert (Merck CO.), measured in a Gemisdch from methanol and 6%iger oaiumacetatlösu g (60: 40 V/V} +2 Avicel SF) (F C CO.); Formic acid Essigsäureasser (27: 75: 900 V/V); 800 V, 15 ia case with the function in accordance with stage B the PEP Amin.Hydrochlorid by 3 (Di-n-butylamino) - propylamin or n {{3 [[N-methyl-n {3 - [2 (p-chlorphenyl) - ethyl] - amino} - propyl]] - amino}} - propylamin one replaces, the appropriate derivatives of the Deamidobleomycin APMP are received. C1 g of the copper-free dAPMP PEP received in accordance with stage B gradates solved in 100 ml methanol, on which the received solution first 700 mg (about 4 equivalents related to Bleomyein) m to adsorb p-Dibenzyloxy-benzaldehyd and then 46 mg (about 1.3 equivalents related to Bleomycin) Cyanoborhydrid added and the reaction mixture after the shifting during 48 h with 40°C into one also - p z0 Cm-Toyopearl () 650 (well - type, manufactured of Toyo soda MFG), which with methanol had before been washed, filled column of 100 ml Fassungsraum one poured, in order the desired connection at the column filling. The column was then washed with 200 ml methanol and eluiert then with linear rising density gradient, by to 500 ml a mixture from methanol and water (7 - “3) continuously one at sodium chloride 0,2molare solution of methanol in water (7: 3) in an l Ienge was added of 500 ml. With a common salt concentration of for instance 0,12molar in a quantity of 130 ml eluierten, blue colored parliamentary groups became into one with Amberli%e (XAD-2 filled column of 160 ml volumes poured, on which the column was eluiert according to a linear density gradient method, as ii methanol was added constantly under linear increase of the methanol concentration to I I a pH value Z0 of 4,5 possessing 0,05molaren buffer solution from acetic acid and Natriumacetat. With a methanol concentration of approximately 9070 eluierten blue parliamentary groups in with amberlites () were poured XAD-2 filled column by i00 ml Fassungsraum, in order to adsorb the desired connection. The column became after washing with 200 ml water with one in the VOL around relationship of i: 4 from 1/50 n-hydrochloric acid and methanol existing mixture eluiert, whereby z5 blue colored parliamentary groups were caught. These parliamentary groups were combined with one another, neutralized with Dowex () 44 (OH-type, manufactured of Dow Chemical Co.) and evaporated to dry ones, with which 670 mg of a blue powder became to receive, which was loosened in 50 ml water. The received solution was poured in with amberlites () XAD-2 filled column of 100 ml Fassungsraum, in order to cause adsorption. The column was then washed to an aqueous 5% solution in the following order with 300 ml by EDTA.2 well, i00 ml one 2%igen aqueous sodium chlorid solution and 200 ml water. The column became then with one in the Volumsverhaltnis of i: 4 from more aqueous 1/50 n-hydrochloric acid and methanol existing mixture eluiert, whereby a maximum adsorption with a wavelength by approximately 290 Nm parliamentary groups showing were caught. These parliamentary groups were stopped using Dowex () 44 (OH-type) to a pH value 5 of 6,0 and gefriergetroeknet then, with which 640 mg of the copper-free hydrochloride became to receive from dDD PEP in a yield from 64% as white amorphous powder. This connection showed an ultraviolet absorption maximum measured in distilled water (E 1%/1 cm) with 285 Nm (74,4). This connection showed measured infrared absorption maxima (Wellenzahl, cm-i) in the KBr tablet with 3400, 2900, 1710, 1650, 1545, 1505, 1450, 1380, 1260, 1135, 1060, 1010, 800, 730 and 690. Further physically chemical characteristics are shown in table I. If in stage C as Bleomycinverbindung dAPMP OCT used wuren, in the course one of the functions similar function dDD OCT described above was received. If in stage C (1) those which can be manufactured the connections appropriate and in accordance with stage, B received (Deamidobleomycin APMP) - derivatives as basic materials were used, (2) as aldehyde the connection appropriate aldehyde which can be made, u.zw. Cyclooctylaldehyd, p-Chlorbenzaldehyd and/or p-Cyanobenzaldehyd were used, (3) these aldehydes in a quantity of 1,2 mol per mol (of Deamidobleomycin APMP) - derivative were used and (4) the conversion according to the above function was made, the appropriate connections indicated in table I were received.