FR2622195A1 - NOVEL ISOPOLYPEPTIDES, PROCESS FOR THEIR PREPARATION AND MEDICAMENTS CONTAINING SAME - Google Patents

Abstract

The invention relates to novel isopolypeptides of general formula I: (CF DRAWING IN BOPI) as well as their salts, their antipodes and their optical isomers. In addition, the invention relates to a process for preparing these compounds. The compounds of general formula I exert an anti-tumor effect and an antiviral effect.

Description

The present invention relates to novel acid isopolypeptides

  diamino-monocarboxylic acids, their salts, their optical isomers and their racemates. In addition, the invention

  relates to a process for preparing these compounds.

  More particularly, the invention relates to novel diamino-monocarboxylic acid isopolypeptides represented by the general formula (1):

R R6 R / R

H HIN-C C C CO CO OH

  I, R15, R, 14H; Wherein R4, R5, R7, F, R, and R10 independently of one another are hydrogen or C1-4alkyl (hereinafter lower alkyl), r is a mean value between 10 and 400, at is an integer of 0 to 10, and

n is an integer from 0 to 10.

  The meaning of the corresponding substituents in

  individual members may be the same or different.

  It is known that isopolypeptides of diamino

  Monocarboxylic compounds rarely appear in nature: in certain peptide antibiotics, in the bacterial cell wall, in the crosslinking of several proteins and in clavicepamines. Some of them have a biological effect and can be produced by microbiological methods as well. According to Japanese research, an L-lysine polymer consisting of 25 to 30 monomeeric units associated with each other via isopeptide bonds is produced by

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  Streptomyces albulus [Shima and Sakai: Reg. Biol. Chem., 45, 2503 (1981)] This poly-L-lysine inactivates various bacteriophages [Shima et al .: Biol., Chem., 46, 1917 (1982)] and a biosynthetic means has been applied for its production. Sima et al .: Nippon Nogekagaku Kaishi 57, 221-226 (1983).) Clavicepamines are lysine-rich base proteins isolated from the saprophytic ergot (Claviceps purpurea) culture and of human pharmocological significance. These proteins could be fractionated into compounds of MW 2,000-17,000 D with a high lysine content between 30 and 95%, and biological studies have shown that they have a retarding effect on cell proliferation, so that these products inhibit growth of animal tumors, practically

  without toxic side effects (Tyihak: Dissertation, 1978).

  From structural determinations, it has been demonstrated that e-lysine (poly) peptides are the basic structural units of clavicepamines [Szokan, Kelemen and T6rôk: J. Chromatogr. 366, 283 (1986)]. The role of structural units of polylsolysin in the effect is not yet known

  biological promise of clavicepamines.

  The antiviral activity of synthetic poly-L-isolysines, their acylated derivatives with amino acids, and poly-p-L-x, pdiaminopropionic acid was measured by Indian researchers. The antiviral activity of these polypeptides has been found to be associated with their ability to induce interferon-like antiviral factors in CRA cultures [Indian

  J. Exper. Biology 20, 227 (1982); Indian J. Chem 218, 483 (1982)].

  In order to study the relationship between the chemical structure and the biological effect, the object of the invention was to produce L-diamino-monocarboxylic acid isopeptides having given PM and low polydispersion, consisting of 10 to 400 units, on an industrial scale too. It is also necessary that the products created have a high purity, a low polydispersion

  and a high stereochemical purity.

  It is well known that the effect of biopolymers is

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  strongly influenced by the distribution of their molecular weight.

  The configuration of amino acids is very important because the different optical isomers can have opposite biological effects and quite other; for example, growth of an animal tumor is inhibited by D-arginine, and stimulated by L-arginine [Szende and Tyihak: Lancet 7546, 624 (1968); Szende: Dissertation, 1974]. Racemic mixtures or racemized polymers do not have the interesting biological effects of origin. These polypeptides had to have both high purity and low polydispersion. The synthetic method has been chosen to solve this problem. An active lysine -Z-protected ester (OPcp) was polymerized according to the method of Hull [Biopolymers 17, 2427 (1978)] and an o-BOC-protected lysine pentachlorophenyl ester according to the procedure of Gangopadhyay and Mathur.

  [Indian J. Chem. 218, 483 (1982). According to the method of Nishi [Int.

  J. Biol. Macromol. 2, 53 (1980)], α-Z-Lys-OH was polymerized by diphenylphosphorylazide (DPPA). Authors of previous articles have not published any data on PM, PM distribution and optical purity of their products. The Applicant's experiments have shown that only low (maximum 2500) and high polydispersion products were obtained with low yields, without any biological effect. The synthetic methods of Kushawa et al. [Biopolymers 19, 219 (1980)] and Mathur et al. [Int. J. Peptide Prot. Res. 17, 189 (1981)] seemed best able to provide low dispersion products. They synthesized a c-poly-lysine and. a-poly-ornithine. The principle of the method was the same for both components: the methyl ester of a protected amino acid was reacted with a protected active ester or an azide to first obtain a tripeptide methyl ester. After alkaline hydrolysis, the tripeptide-acid was activated into an ester form

  active (it was converted to its pentachlorophenyl ester).

  After deprotection of the w-terminal amino group, the tripeptide was ready and active for polycondensation. Finally, the -amino groups

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  protectors were removed and the isopeptide isolated. In these procedures, protective amino groups BOC at position a and Z at position w were used. Their removal was carried out by treatment with a strong acid and by catalytic hydrogenation respectively. In the above publications, the polymers are not characterized by PM, PM distribution or

optical purity data.

  The abbreviations recommended by the

  IUPRC-IUB Commission on Biochemical Nomenclature, J. Biol.

  Chem. 247, 977 (1972), as well as a few other names, some of which are given below: Z benzyloxycarbonyl BOC t-butoxycarbonyl-N-pnitrophenyl-O-N-nitrosuccinimidyl-OPcp pentachlorophenyl-OPfp pentafluorophenyl-O-B-N-hydroxy-benzotriazolyl BuOCO isobutyloxycarbonyl EtCOO ethyloxycarbonyl N3 azido EEDQ Nethoxycarbonyl-2-ethoxy-1,2-dihydroisoquinoline EtORc ethyl acetate IIDQ N-isobutoxycarbonyl-2-isobutoxy-1,2-dihydroisoquinoline THF tetrahydrofuran DMF dimethylformamide HPLC high performance liquid chromatography OPTLC layer chromatograph Thin δ overpressure IEF electrofocusing MS mass spectrometry NMR nuclear magnetic resonance spectroscopy ZCO2 Z group measured from CO 2 released PM TLC molecular weight thin layer chromatography

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Butanol DuOH

RcOH acetic acid.

  Several disadvantages have been observed and experienced in reproducing these processes: to prepare oligomers, their carboxy groups were always protected by a methyl ester, so that the oligomer obtained was not directly usable for polycondensation. To obtain an active intermediate, the carboxy group had to be liberated by subjecting the corresponding methyl ester to alkaline hydrolysis. This step results in racemization and other side reactions, which decreases the yield. Unfortunately, the synthesized polymers also exhibited high polydispersion. In addition, the application of a benzyloxycarbonyl protecting group for a β-amino function was disadvantageous because of its elimination by catalytic hydrogenation. Unfortunately, the polymers thus prepared were

  ineffective in the biological tests of the Applicant.

  The object of the invention is to eliminate the disadvantages mentioned above by the development of a new process, which has never been applied in the production of diamino-monocarboxylic acid isopeptides. The invention is based on the finding that the above object can be achieved if directly applicable active oligomers for polycondensation are first prepared in a process, wherein the carboxy groups are protected by active ester groups and coupling peptides is carried out at a much greater rate than the aminolysis of the protective active ester group. This coupling method provides a

  degree of activation greater than that of the active ester method.

  It has been found that the "release" activation method is well applicable for preparing isopeptides and diaminocarboxylic acid salts. A p-nitrophenyl ester should be used for the temporary protection and simultaneous activation of COOH groups, and the mixed anhydride method for

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  peptide coupling, with BOC protection for amino groups in the last amide linkages and with a Z group for aoino groups not taking part in the reaction. It has been established that in this way, homo-isopeptides or isopeptides can be synthesized in sequence of any diamino carboxylic acid. In addition, it has been found that low dispersion isopeptides can be obtained reproducibly with P1 within wide limits. Intermediate and final products are well crystallizable substances of high purity and

  they can be easily isolated.

  In addition, the invention is based on the finding that p-nitrophenyl esters are the most favorable groups for temporary protection and simultaneous activation of carboxy groups in the case of diaminocarboxylic acids, the mixed anhydride method is optimal. here for peptide coupling, and Z and BOC groups can

  to be chosen for the protection of others.

  Finally, the invention is based on the finding that the homoisopeptides or isopeptides in sequence of any diaminomonocarboxylic acid can be synthesized by the above method without racemization, i.e. with

conservation of their configuration.

  From the above, the invention relates to isopolypeptides, their salts, their optical isomers and their racemates having the general formula (1), in which the

  substituents are as defined above.

  According to the present invention, the compounds of general formula (1) and their salts are prepared by coupling of a monomer or an oligomer of general formula (1)

4 6 1

R5 R7n NRR n (z

7 9

L - 21HR q

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  in which q = 1 to 9, R 1 is an amino protecting group (in oligomers, all R 1 are the same), R 3 is an active ester group for carboxy groups suitable for protection and simultaneous activation, R 4, R5, R6, R7, R8, R9, R10, a and n are as defined above - with a monomeer of the general formula (III) R4 / R1 R / R8 \ R2 1 (1-C m-A, 7Ox in which R2 is an amino protecting group different from R1, which can be selectively removed from R1, X is a carboxy activating group provided that the -COX group is more active than the COOR3 group, R1, R4, R5, R6, R7, R1, R9,

  R10iO met n are as defined above.

  The protecting group R2 is removed from the compound of general formula (IV) thus obtained

R4R R / R

2 1 1 10 1R.

R, HN .. - CO OR

R 15V7 1 V1OR (IV)

7

R R 1NHR R

  in which the substituents are as defined above and s is an integer of 2 to 10 and then, if s, the resulting compound

2 6 2 2 1 9 5

  is treated with a compound of the general formula (III) or is polycondensed in a known manner. The protecting groups R 1 are then removed from the products. If the obtained compound of general formula (1) is in the free base form, it can be converted into a pharmaceutically acceptable salt, or if it is a salt, it can be converted into a free base or a

other salt.

  Optical isomers can be prepared using the corresponding isomers as starting materials. If racemic polymers are to be prepared, the racemates of the

  monomers must be used as starting materials.

  In the process of the invention, R 1 and R 2 are selectively removable protecting groups well known in peptide chemistry. R 1 is preferably benzyloxycarbonyl (Z) and R 2 is preferably t-butyloxycarbonyl (BOC). The group -OR3 is an active ester group for both protection and activation, for example -ONp, -OSn, -OPcp, -OPfp, -0Bt; X can be BuiOCO-, EtOCO-, N3, -OSn, -OPcp, -OPfp, depending on the reaction conditions and the

  meaning of R3, for example it is a group BuiOCO-, EtOCO.

  According to the most advantageous embodiment of the present invention, in the monomer of general formula (11) used as starting material, R1 is Z, R2 is BOC and R3 is -ONp. The BOC group is removed by acidolysis, and then the protected diaminocarboxylic acid (R1 = Z, R2 = BOC) is coupled according to the mixed anhydride method. After removal of the BOC protecting group, the dipeptide thus obtained is polycondensed or it is coupled again with the protected amino acid according to the above, the

  steps being repeated until a decapeptide is obtained.

  The best method is to repeat the couplings up to the tripeptide and transform the resulting oligomer directly suitable for polycondensation into an isopolypeptide having the expected number of amino acid residues. Removal of the protecting group Z (i.e. deprotection) is preferably carried out by HBr in acetic acid, and the final product is

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  The conditions of the most advantageous procedure of the mixed anhydride coupling method are as follows: a) solvent: acetonitrile, THF, DMF, dichioromethane; b) anhydride-forming reagent: ethyl chloroformate, isobutyl chloroformate, EEDQ, IIDQ; tertiary base: triethylamine, diisopropyl-ethylamine, N-methyl-morpholine; c) activation time and temperature: 10 to 30 minutes, between -10 C and -20 C; d) neutralization of the salt of the amino component:

  with triethylamine or diisopropyl-

  ethylamine in the reaction mixture.

  Yield in pure end products: 60 to 0% X.

  Both homo-isopolypeptides and isopeptides

  in sequence can be prepared by the process of the invention.

  Current separation techniques, such as HPLP, OPTLC, IEF and structure determination methods such as IR, MS, NMR, have been used to analyze prepared isopeptides. The products studied and the witnesses put the

structures in evidence.

  Substances purified as described in

  examples were used for the study of biological activities.

  Purification was carried out by precipitation, chromatography

on column and on gel.

  From the biological studies, it has been found that among the poly-Lisolysins (degree of polymerization (dip) of 10-400) prepared by the process of the invention, the polymers having a d.p. from 40 to 200 (Pl 5,500 - 20,300 D) have a retarding effect on cell proliferation, i.e. anti-tumor activity in vitro and in vivo, and they inhibit the formation of tumor metastases. Products with a MW of 10.200 to 15.400 D (from 80 to 120) have the highest activity. Other substances prepared until now were found to be ineffective, mainly because of a strong

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  Polydispersion of the final products and racemization during the

  production in the Kushawa or Mathur methods.

  It has been found that poly-L-lysines having a d.p. from to 120 produced by the Kushawa or Hull method, do not exhibit antiviral activity. The main advantages of the synthesis of the present invention over the known procedures are the following: - Products having a promising biological effect can be

  obtained with high reproducibility.

  - Applicable oligomers for the polycondensation can be obtained, so that the hydrolysis step mentioned above is eliminated and the racemization which is very

  dangerous from the point of view of the biological effect, can be avoided.

  Products with a favorable PM distribution (i.e., low polydispersion) can be obtained between wide MW limits, which is also advantageous for having the desired biological effect (eg in the case of SZTP-14 PM polymer: 12,700,200; polydispersion: 3.7). The yields are higher,

  the purity of intermediate and final products is greater.

  These two substances are well crystallizable and they can be easily isolated. The reproduction of the steps does not pose any difficulties. - The peptide coupling steps require durations of

shorter reaction.

  - The deprotection by catalytic hydrogenation can be eliminated

  by the combination according to the invention aminoprotective groups.

  This implies other advantages in the production on an industrial scale. The greatest advantage of the process of the invention is the

  performing the reaction steps without racemization.

  As mentioned above, the products obtained until now have proved ineffective in the tests performed by the Applicant. On the other hand, polymers synthesized by the process of the invention have

very promising biological

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  Among the compounds of general formula (I) prepared according to the process of the invention, the following compounds in which the substituents are as defined below are new: R4, R5, R6, R7 and R10 represent independently the each other, a hydrogen atom or a methyl, ethyl, propyl or butyl group, R8 and R9 are each a hydrogen atom, r is an integer of 10 to 400, a is 0, 1, 2 or 3, and n is 0. In a group of preferred compounds: R4, R5, R6, R7 and R10 independently of one another are hydrogen, methyl, ethyl, propyl or butyl, R8 and R9 are each a hydrogen atom, r is an integer of 40 to 200, is 3, and n is 0. In another group of advantageous compounds: R4, R5, R6, R7 and R10 each represent a hydrogen atom, R8 and R9 are each a hydrogen atom, r is an integer of 80 to 120, a is 3, and n is 0. In another group of advantageous compounds: R4, R5, R6, R7 and t RO10 represent independently of each other a hydrogen atom or a methyl, ethyl, propyl or butyl group, R8 and R9 are each a hydrogen atom, r is an integer of 10 to 120,

262 2 1 95

  to 2, and n is 0. In another group of preferred compounds: R 4, R 5, R 6, R 7 and R 10 independently of one another are hydrogen, methyl, ethyl, propyl or butyl, R 8 and R9 are each a hydrogen atom, r is an integer of 10 to 400, is 1, and

n is 0.

  In a group of other advantageous compounds: R4, R5, and R10 independently of one another, a hydrogen atom or a methyl, ethyl, propyl or butyl group, R6, R7, R8 and R9 are each an atom of hydrogen, r is an integer of 10 to 400,

m and n are 0.

  The L stereoisomers of the isopolypeptides of general formula (1) and their salts have proved very effective against

  tumors and viral infections.

  The biological activity of the polyisolysins prepared by the method of the invention has been studied in vitro and in vivo. Promising activities are highlighted in the next section. Anti-tumor activity was measured on animal tumor strains and antiviral activity was studied against

plant viruses.

  It was found that in the experiments carried out, the isopoly-L-lysines having a P1 of 5,500 to 25,600 had a surprising activity, among which the compounds having a P1 of

  10,200 to 15,400 were the best.

  SZTP-14 was the most effective substance in

two measures.

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  It was characterized as follows: ## EQU1 ## The viscosity of the Z-protected polymer: = 31.92 cw / g ZCOO 2: 16.8% (calc.), 16.3% (found).

  The free polymer was available as hydrobromide (HBr).

  Br content: 37.7% (calcI), 36.8% (found).

  R254 = 0 (i.e., no UV absorption).

  It was purified by precipitation in alcohol with

ether.

[f] 20D = +32.4 (c = 2; water)

Pl = 12,700 D 200.

  Study of anti-tumor activity The effect of compounds that can influence cell proliferation is generally studied in vitro with cell cultures, and in vivo with transplantable animal tumors. Thus, the compounds according to the invention were tested on a human erythroleukemia cell line (K-562, Karolinska Institute, Stockholm) and on four transplantable mouse tumors (lymphoid leukemia L1210, tumor derived from

  macrophage P380, Ehrlich carcinoma, Lewis lung tumor).

  In the case of L1210 and P380 tumors and Ehrlich carcinoma, animal survival was used as an indicator of the effect. In the case of LLT, the number and size of metastases were determined in the lung or liver of treated and untreated mice. The experiments were carried out with the following injection solution: 50 mg of the compound dissolved in 10 ml of

physiological saline solution.

  The compounds studied were designated as follows: SZTP: polyiso-L-lysines produced by the process of the invention SZTP-14: polyiso-Lysine hydrobromide, P1 calculated on free bases: 12.700 200, polydispersion: 3.7 ( number average / weight average) SZTP-15: polyiso-Lysine hydrobromide, P 'calculated on bases

26 2 2 1 9 5

free: 11,600,200.

  SZTP-16: polyiso-L-lysine hydrobromide, PM calculated on bases

free: 13,400,200.

  SZTP-17: polyiso-L-lysine hydrobromide, PM calculated on free bases: 14,500 200. SZTD: polyiso-L-ornithine produced by the process according to the invention SZTD-18: polyiso-L-ornithine hydrobromide, PM calculated on

free bases: 8,900,200.

  H-17: polyisolysin produced according to Hull et al.

  K-17: polyisolysin produced according to Kushawa et al.

A-3: c-polylysine.

  Summary of Experiments on the Proliferation of Tumor Cells The compounds produced according to the invention reduced in vitro, depending on the dose, the number of tumor cells relative to that of the control. This effect resulted in a stagnation of the number of cells in the case of low doses, high doses causing cell death. H-17 and K-17 were ineffective and m-polylysine (R-3) only caused stagnation

the number of cells.

  According to our in vivo experiments, treatment with SZTP resulted in increased survival of the treated animals. In the case of the Ehrlich tumor, the treatment caused a very long tumor-free period and the growth of

  tumor was observed only after stopping treatment.

  Treatment with SZTP completely prevented the formation of metastases in the liver when the tumor was inoculated into the spleen and decreased the number and size of lung metastases when the tumor was inoculated into the muscle. As a result, the polyisolysins and polyisoornithines produced according to the invention have been found to be inhibitors of tumor cell proliferation. Among these compounds, poly-isolysines exhibited a surprisingly strong inhibitory effect. This effect can be easily compared to the effect produced by the best known cytostatic drugs with respect to animal survival. The surprisingly good antimetastatic effect of the compounds of the invention is obvious. The compounds of the invention have the additional advantage that they exhibit low toxicity, which is much lower than that of

  cytostatic drugs applied until now.

  Biological studies of the Applicant have shown maximum tolerable dose for example that the minimum effective dose therapeutic index has a value greater than 10, while this value is generally well below 10 in the case of most

known cytostatic drugs.

  There was no difference between the biological response of males and females in this respect, which is also

an advantage.

  The following data illustrate the anti-tumor effect.

  In Vitro Experiments Tables 1a to 1c show the effect of the polyisolysins produced by various processes, polyisoornithines produced by the process of the invention and polylysine on the

proliferation of K-562 cells.

  The experimental parameters were as follows: Cells: human erythroleukemia cell line K-562

  (Karolinska Institute, Stockholm) was used.

  Number of cells: 5x104 / ml, in glass test tubes

containing 5-6 mI of medium.

  Hombre of probes: 3 test tubes per dose.

  nilieu: Parkers n-199 supplemented with 10% X fetal bovine serum (Flow

Laboratories, Irvine, Scotland).

  Temperature, atmosphere: 37 C, 5% C02 + 95% air.

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Dose: 1-10-100 μg / ml.

  Evaluation: cell count 24, 40, 72 and 96 hours after

  dilution of cultures, with a Buerker chamber.

Number of experiments: 2 / compound.

  The Table shows that treatments with SZTP-14, SZTP-15, SZTP-16, SZTP-17 and SZTP-18, but especially SZTP-14, resulted in a significant dose-dependent inhibition of cell proliferation. . It is remarkable that even treatment with 1 μg / ml caused a well-defined antiproliferation effect. Table lb, however, shows that, on the contrary, H-17 and K-17 had no influence on cell proliferation.

K-562.

  Table 1c shows that polylysine caused the stagnation of the number of cells compared to that observed in the control, but does not cause a decrease in the number of cells.

  number of cells even at a high dose (100 μg / ml).

  The values shown in the Tables indicate the number of cells evaluated at given moments in time. The cells were counted in 3 test tubes for each of the doses and in 3 test tubes as a control at each moment

in time.

  Table II shows the effect of the compounds prepared according to the invention (SZTP-14-15-16-17-18) on the K-562 cell line,

in vitro. Treatment: 1-10-100 μg / mg, 24 hours after dilution of the culture.

  Tabl water Number of cells x 103 / nutrient liquid mel SZTP-14 48h 72h 96h 100 / ug / ml 0 0 0 / ug / ml 90 90 150

80 120

80 140

1 / ug / ml 80 170 180

180 190

120 150

Witness 130 220 260

230 230

110 220 230

SZTP-15

/ ug / ml 40 10 10

20 10

10 20

10 / ug / ml 80 100 160

120 110

100 140

1 / ug / ml 120 150 200

140,190

120 120 210

Witness 140 210 270

190,250

190,280

Table la (continued) 48h 72h 96h

SZTP-16

/ ug / ml 100 160 160

120 120 140

140 140

/ ug / ml 120 160 220

160,240

180,240

1 / ug / ml 140 180 260

210,280

240 250

Witness 140 220 280

200,260

140,200 270

SZTP-17

/ ug / Ml 110 180 200

190,210

200,200

10 / μg / ml 120 200 260

200,260

190,280

Witness 150 260 280

210,280

130 210 260

Control 140 250 280

220 280

220,260

Table the (end) 48h 72h 96h

SZTP-18

/ ug / ml 100 160 180

130 180

130,160

10 / ug / ml 120 180 220

160,230

190,230

1 1 1ug / Ml 140 200 280

210,260

220 280

Witness - 140 220 280 Witness

230 280

* 230 270

Table lb

  It shows the effect of treatment with H-17 and K-17 on

  proliferation of K-562 cells in vitro.

  Treatment: 1-10-100 μg / ml, 24 hours after dilution of the culture. Tabl water lb Number of cells x 103 / ml of nutrient liquid 48h 72h 96h

H-17

/ ug / m1 125 230 280

210,270

210,280

/ ug / ml 120 220 290

240,280

210,220

1 / ug / ml 130 240 290

240,290

220,270

K-17 100 / ug / ml 120 220 280

210,290

220,270

/ ug / ml 125 240 290

240,290

210,270

1 / ug / ml 140 240 290

220 280

230 280

Witness 120 210 290

200,280

120,220,285

Table lc

  It shows the effect of R-3 treatment on the

K-562 cell line, in vitro.

  Treatment: 1-10-100 μg / mg, 24 hours after dilution of the culture.

Table 11

  Number of cells x 103 / ml of nutrient liquid 48h 72h 96h / ug / ml 110 110 100

120 110 100

120 110

/ ug / ml 130 120 120

110 110

120 120 110

1 / ug / ml 110 110 100

110 110

120 110

Witness 120 250 300

240,290

125,240,290

  In vivo studies a) Tumor L1210, study of SZTP-14

  Origin of the tumor: Chester Beatty Institute, London.

  Animal strain: DBR / 2 consanguine, personal breeding.

  Weight and sex of animals: 20-23 g, females, 5 animals per group. Number of tumor cells: 105 per animal, intraperitoneally. Treatment: 50 mg / kg, intraperitoneally, 24 hours after inoculation of the tumor, once daily during

8 days.

  Control group: physiological saline solution, 0.2 ml, intraperitoneally.

Evaluation: survival of animals.

  Results: As shown in Table 2, the survival of the treated animals exceeded that of the controls. Ten days after inoculation of the tumor, no mice in the control group were alive. At the same time, all the treated animals were alive. The first treated animal died 13 days and the last 15 days after

Inoculation of the tumor.

Table 2

  Effect of daily treatment at 50 mg / kg on survival of DBR / 2 mice inoculated with L1210 tumor Days after inoculation Live controls Live treated mice

9 3 5

- 5

11 - 5

12 - 5

13 - 4

14 - - 3

-

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  b) Ascitic tumor P-366, treatment with SZTP-14

  Animal strain: hybrid BDF / 1, personal breeding.

  Weight and sex of animals: 20-23 g, males, 5 mice per group.

  Number of tumor cells: 2.8 × 10 6 per animal, intraperitoneally. Treatment: 10 mg / kg, intraperitoneally, it started 24 hours after tumor inoculation, once a day. Control group: 0.2 mi of physiological saline, once

  per day, intraperitoneally.

Evaluation: survival of animals.

  Results: Table 3 shows the number of surviving animals.

  All the treated mice survived the animals of the

a group of witnesses.

Table 3

  Effect of daily treatment with 10 mg / kg SZTP-14 on the survival of BDF / 1 mice inoculated with ascitic tumor P-388 Days after inoculation Live controls Live treated mice

9 4 5

10 - 5

ll - 5

12 - 5

13 - 5

14 - 5

15 -

16 - 2

17 - 1

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  c) Ehrlich ascitic tumor, treatment with SZTP-14

  Animal strain: Swiss, not consanguineous, animal husbandry.

  Weight and sex of animals: 20 9, 5 males and 5 females per group. Number of tumor cells: 106 per animal. Treatment: 50 mg / kg and 75 mg / kg, intraperitoneally, it started 24 hours after inoculation of the tumor,

once a day for 20 days.

  Control group: 0.2 ai physiological saline,

intraperitoneally.

  Assessment: measurement of body weight, every day of survival. Live animals after tumor inoculation were killed, autopsied, the amount of ascites was measured and the formation of a solid tumor in

  the peritoneal cavity has been recorded.

  Results: Table 4 shows the body weight of the mice from the 16th day following the inoculation of the tumor. The animals in the control group died between the 17th and the 22nd day. Excess weight indicates the formation of tumor ascites. In each treated group, the animals were alive (except for one group) 55 days after inoculation of the tumor. The animals were subsequently killed. Table 5 shows the amount of ascites and the presence of solid tumor in the peritoneal cavity. This means that no tumor-free animals were found, but on treated mice, 16 were alive 33 days later.

  the death of the last mouse of the control group.

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Table 4

  Effect of intraperitoneal treatment with 50 mg / kg and 75 mg / kg SZTP-14 (for 20 days) on body weight and survival of male and female Swiss mice inoculated with an Ehrlich ascitic tumor. Body weight before inoculation of the

tumor: 20 g.

  0 = death due to the tumor; + = dead for other reasons.

  Day after inoculation Witness Control o 75 mg / kg

  of tumor 1. 2. 3. 4. 5. 1. 2. 3. 4. 5. 1. 2. 3. 4. 5.

  16. 43 37 41 46 40 40 34 40 35 41 17 22 24 21 22

  17. 44 39 42 46 41 40 35 0 35 42 17 23 24 25 22

  18. 45 42 43 47 49 42 35 0 35 43 18 23 24 21 24

  19. 0 43 43 50 0 0 36 0 0 44 19 23 24 21 24

  20. 0 46 0 42 0 0 42 0 0 46 16 21 23 20 21

  21. 0 46 0 43 0 47 0 047 18 23 25 25 22

  22. 0 0 0 0 0 0 000 0 51 19 25 26 25 24

24. 0 0 0 00 0 0 0 0 0 1924262425

25. 20 24 26 24 26

26. 20 24 27 24 25.5

27. 21 25 28 25 26

28. 21 25 28 25 26

29. 21 25 28 25 26

33. 22 25 28 25 26

34. 22 25 28 25 26

42. 23 26 27 25 28

50. 24 27 27 25 28

55. 26 29 0 27 32

  Table 4 (end) Day after inoculation 75 mg / kg c? 50 mg / kg 50 mg / kg o

  of tumor 1. 2. 3. 4. 5. 1. 2. 3. 4. 5. 1. 2. 3. 4. 5.

  16. 28 23 28 25 29 18 19 22 23 + 29 30 31 26 27

  17. 28 23 28 24 28 18 19 22 23 + 28 30 31 26 28

  18. 29 24 27 25 30 19 20 23 23 + 29 30 32 27 29

  19. 29 24 28 26 30 20 20 23 23 + 29 30 32 28 29

  20. 27 22 25 24 27 17 18 21 21 + 27 28 30 24 27

  21. 29 23 27 27 29 18 18 22 22 + 27 30 29 25 27

  22. 29 24 28 27 30 20 20 24 22 + 29 30 32 25 29

  24. 29 25 29 28 31 2020 24 23 + 29 30 32 26 30

  25. 30 26 30 29 31 21 21 24 24 + 30 30 32 26 31

  26. 30 25 30 29 31 22 22 24 25 + 30 31 32 26 31

  27. 30.526 31 30 31 22 22 25 25 + 30 31 32 25 32

  28. 31 26 31 30 31 22 22 25 25 + 30 31 31 25 32

  29. 31 26 31 30 31 22 22 25 25 + 30 31 32 26 32

  33. 30 27 32 31 32 23 22 27 26 + 28 32 30 24 33

  34. 30 27 32 31 32 23 22 30 26 + 28 32 30 24 33

  42. 32 29 33 34 28 24 25 30 31 + 27 33 30 26 35

  50. 34 30 35 37 24 24 26 30 36 0 34 30 26 36

  55. 33 32 38 0 37 26 28 32 43 + 0 36 31 27 38

2622 1 9 5

Table 5

  Rscites and solid tumor in the peritoneal cavity of Swiss mice inoculated with an Ehrlich ascitic tumor, treated with 50 mg / kg and 75 mg / kg of SZTP-14 and killed 55 days after tumor inoculation. Treatment mg / kg 75 mg / kg 50 mg / kg 50 mg / kg

Q 09 0

  Rscites mli - - 0 - 4 1 6 - O- - 5 2 4 0 - 1 -4 0 Solid tumor in the cavity + + 0 + + +++ 0 + + + + O. + 0 abdominal O: the animal was dying earlier

  +: solid tumor in the abdominal cavity.

  Inhibition of tumor metastasis with SZTP-14 a) Lewis lung tumor (LLT), inoculation in the spleen

  Animal strain: Inbred C57B1, LRTI, Gôdôllo, Hungary.

  Weight and sex of the animals: 20 to 23 g, females (6 witnesses and 4

treated).

  Number of tumor cells: 5x106 LLT cells inoculated into the spleen. Treatment: 75 mg / kg, for 8 days, it started 24 hours

after inoculation of the tumor.

  Assessment: Animals were killed and liver metastases were counted 9 days after tumor inoculation. Results: Table 6 shows the number of metastases. No metastases were found in the liver after

treatment with SZTP-14.

2622 1 9 5

Table 6

  Effect of treatment with SZTP-14 (75 mg / kg intraperitoneally for 8 days) on the number of liver metastases in C57B1 mice inoculated with LLT in the spleen Number of liver metastases Control 31 20 63 36 38 56 (x = U0.66, s = 16.02, n = 6) SZTP-14 0 0 0 0 (n = 4) b) Lewis lung tumor (LLT), inoculation in the muscle of the right thigh

  Animal strain: Inbred C57B1, LRTI, G6dollo, Hungary.

  Weight and sex of animals: 20 to 22 g, females.

Number of animals: 5 per group.

  Number of tumor cells: 5x105 cells / animal intramuscularly in the muscles of the thigh of the right lower limb. Ten days after the inoculation of the

  tumor, the affected end of tumors was removed.

  After the operation, between the 11th and 17th days following the inoculation of the tumor, the following treatment was administered: 50 mg / kg of SZTP-14 per day intraperitoneally. Controls were administered 0.2 ml of physiological saline daily intraperitoneally. Evaluation: The animals were killed on the 18th day after inoculation of the tumor. The average number and volume of lung metastases were determined by stereomicroscope. Results: Table 7 shows the average number and volume of lung metastases. Treatment with SZTP-14 significantly reduced the mean number and volume of metastases

lung.

2622 19 5

Table 7

  Effect of treatment with SZTP-14 (50 mg / kg daily, intraperitoneally between days 11 and 17 after tumor inoculation) on the mean number and volume of lung metastases in C7B1 mice inoculated with intramuscular

with a Lewis lung tumor.

  Treatment Number of metastases Average number of metastases (mm3) on average 63.13 + 27.53 49.45 29.07

SZTP-14 26.29 + 18.87 20.10 35.38

  Demonstration of the antiviral effect Reduction in the synthesis of the tobacco mosaic virus

  Intact tobacco plants (Nicotiana tabacum cv.

  Samsun), susceptible to infection by tobacco mosaic virus (TMU), were mechanically inoculated. Carborundum powder (particle size: 500 mesh) was used as an abrasive. The inoculum contained 200 pig of purified tobacco mosaic virus TMi (strain TMilU U1) in 0.1 M Sørensen phosphate buffer (pH 7.2). Intact leaves were treated with a solution of SZTP-14 as a foliar spray 3 days after inoculation. The treatments were repeated for three consecutive days. The severity of symptoms was assessed two weeks after inoculation. Leaf samples were also taken 11 days after inoculation from all the leaf sequences of inoculated plants, ie leaves.

  different heights and ages.

  The virus concentration was measured from one gram of the frozen samples. Leaf samples were homogenized in 4 ml phosphate buffer (pH 7.2) and centrifuged in an Eppendorf centrifuge (approximately 10,000 rpm).

2622 1 9 5

  for 5 minutes). Fifteen ul of supernatant was used for quantitative measurements. Rocket immunoelectrophoresis was used to estimate the virus content with specific anti-TlU (U1) sera (1 μl immunoglobulin / 1 cm). Sodium barbiturate buffer (0.2 M, pH 6.6) was used as the current buffer. The virus content was calculated from the height of the precipitation peaks, with a standard calibration curve.

Table 8

  Example for the estimation of the antiviral effect Treatment with SZTP-14 Virus content mg / I 11 mg / ml 20 mg / I 24.6 mg / ml 2 mg / 1 29.3 mg / ml Control 51.3 mg The isopolypeptides according to the invention in the form of the free base and the non-toxic salts can be used in

  pharmaceutical preparations as effective substances.

  In this regard, they are mixed with carriers and auxiliary agents commonly used in the pharmaceutical industry and are thus transformed into pharmaceuticals. In addition, the compounds according to the invention can be used both in the form of free base and salt to increase the resistance of plants to viral infection, after mixing with agents.

  auxiliaries commonly used in plant protection.

  The salts which are pharmaceutically acceptable are, for example, acid addition salts such as a hydrochloride, a hydrobromide, a hydroiodide, a sulfate, a phosphate, a nitrate, an oxalate, a fumarate, a gluconate, a tartrate or a maleate. , an acetate, a citrate, a benzoate, an ascorbate, a lactate, an alginate, etc. The pharmaceutical products containing the compounds according to the invention as active agents may be injectable solutions, tablets, dragees, suppositories, suspensions, drinking solutions, powder mixtures, etc. The pharmaceutically effective amount of the compounds according to the invention depends on the mode of application and the condition of the patient. In general, a daily dose of 0.02-20 mg / kg body weight is administered in divided doses in 2 to 4 doses per day. The doses are higher in the case of a

oral administration.

  Advantageous forms of drug are injection solutions that can be administered intravenously, subcutaneously or intramuscularly. To prepare injectable solutions, an isotonic NaCl solution is used, optionally with a phosphate buffer, an ascorbic acid stabilizer, etc. The active substances can be administered in the form of solid and liquid drug preparations. In the case of liquid forms, sweetening agents may be added

and / or perfumes.

  Advantageous solid units to be administered by

  oral tablets are for example tablets, dragees and capsules.

  To prepare these products, the active agent is mixed with pharmaceutical carriers and the mixture is compressed into tablets, converted into dragees or introduced into capsules. The conventional additives and fillers are used as carriers, for example lactose, magnesium stearate, sucrose, talc, stearic acid, gelatin, agar-agar, pectin, gum tragacanth, colloidal silica gel, corn starch, alkenic acid, etc. The cocoa butter is

  advantageously used to prepare suppositories.

  The treatment of the plants is conveniently carried out by conversion of the compounds according to the invention with usual auxiliary agents, such as moistening substances and solid supports, preferably kaolin, in solid or liquid preparations, preferably in combination with other substances.

2622 1 9 5

  active. A solution or a suspension containing the compound according to the invention at a concentration of 1 to 50 ppm is prepared from the preparation thus obtained, and it is applied in a

  known to plants, preferably at 0.5 to 5 g / ha.

  The invention is further illustrated with reference to

following non-limiting examples.

EXAMPLE 1

  Poly-L-Lysine Hydrobromide a) P-Nitrophenyl Ester Hydrochloride

N-α-benzyloxycarbonyl-L-lysine.

  10 g of N-α-p-nitrophenyl ester were dissolved

  benzyloxycarbonyl - (N-E-tert-butyloxycarbonyl) -L-lysine in 40 ml of trifluoroacetic acid and added an equivalent amount of 2N HCl in EtOAc. After standing for one hour at room temperature, the solution was evaporated under vacuum to half volume,

  then ether was added and the solution was crystallized.

  Yield: 8.3 g (96.06 ×); m.p. 83-86 6C; TLC Rf = 0.68

  (n-butanol-acetic acid-water 4: 1: 1).

  b) N-α-benzyloxycarbonyl-N-ú- p-nitrophenyl ester

  (N'-N-benzyloxycarbonyl-N'-s-tert-butyloxycarbonyl -L-

lysil) -L-lysine

  7.69 g (20 moles) of N-α-benzyloxycarbonyl were dissolved.

  (N-e-tert-butyloxycarbonyl) -L-lysine in absolute acetonitrile. 2.8 ml (20 mmol) of triethylamine and then 2.8 ml of isobutyl chloroformate were added dropwise with vigorous stirring. After 20 minutes of activation, 9 g of the salt from step a) were added and then a cold solution of 2.8 ml of triethylamine in 5 ml of acetonitrile was added dropwise. Stirring was continued for 2 hours at -10 ° C. The mixture was diluted with 300 μl of 0.5 M HCl. The solid product was collected, dried and recrystallized from a 1: 1 mixture of ethyl acetate and petroleum ether. yield:

  12.9 g (89.02X); mp: 128-131 C. Analysis: content of -ONp: 99.8X.

  TLC Rf = 0.64 (EtOAc-cyclohexane 3: 1). Rf = 0.91 (n-butanol-acid

2622 1 9 5

  acetic acid-water 4: 1-1); HPLC tr. = 8.8 min. (k '= 2.2); column: ODSHypersil-6 (125x4 ms); eluent: neOH-CH3CN-0.02M NaORc buffer (pH 4) 40:30:30 v / v; flow rate: 1 ml / minute; detection: UU 254 nm. 0c) p-nitrophenyl ester hydrochloride of h-

  c-benzyloxycarbonyl-N- (N '-ac-benzylcarbonyl-L-lysine)

  L-lysine 10.25 g of the salt were prepared from 10.5 g of ester

  N-α-Benzylocarbonyl-N-s- (N'-t-benzyloxy)

  carbonyl-N '-c-tert-butyloxycarbonyl-L-lysine) -L-lysine according to the procedure of step a). M.p .: 109-113 C. TLC Rf = 0.70

(n-BuOH-RcOH-water 4: 1: 1).

  d) N-e-tert-butyloxy-p-nitrophenyl ester

  carbonyl-N-α-benzyloxycarbonyl-L-lysine)

  5.71 g (15%) of N-α-benzyloxy-

  carbonyl - (-s-tert-butyloxycarbonyl) -L-lysine mixed anhydride according to the procedure of step b), with triethylamine, isobutyl chloroformate in acetonitrile. We added 10.5

  N-α-p-nitrophenyl ester hydrochloride

  benzyloxycarbonyl-N-E- (N'-α-benzylcarbonyl-L-lysil) -L-lysine in the presence of triethylamine. 13.9 g of product (85.4% X) were prepared according to the procedure of step b). It was crystallized from acetonitrile and ether. M.p .: 145-150 C. Analysis: 99.6% ONp content; TLC Rf = 0.98 (n-BuOH-RcOH-water 4: 1: 1); HPLC tr. = 18.2 min. (k '= 8.1); column: ODSHypersil-6 (125x4 mm); eluent: MeOH-CH3CN-0.02M NaORc buffer (pH 4) 40:30:30 v / v; flow: 1

ml / minute; detection: UU 254 nm.

  e) Tri-N-p-nitrophenyl ester hydrochloride

  a-benzyloxycarbonyl-L-lysine) 12.2 g of ester prepared in step d) were treated according to the procedure of step a). 12.1 g (95.4X) were thus obtained.

  salt; m.p .: 108-125C. TLC Rf = 0.71 (n-BuOH-RcOH-water 4: 1: 1).

  f) Poly-N-benzyloxycarbonyl-L-lysine 23 g (24 moles) of tri- (N-α-benzyloxycarbonyl-Lysine) p-nitrophenyl ester hydrochloride were condensed in 60 ml.

26 2 2 1 95

  of dimethylsulfoxide in the presence of 4.5 ml of triethylamine until a gel is formed. The polymer was isolated by pouring it into an aqueous solution of 5% H 2 CO 3. Yield: 16.8 g (89.6%). Analysis: ZCO 2: 16.8% (calc.), 16.3% (found); [qsp] / = 31.92 cm3 / g (c = 4.95 g / l, dichloroacetic acid). g) Poly-c-L-lysine brothydrate 16 g of the protected polymer of step f) were dissolved in 100 μl of trifluoroacetic acid and 100 μl of HBr were added

  4N / RCOH. The final product was precipitated in ether.

  Yield: 14.7 g (91.6%). Analysis: Br: 37.7% (calc.), 36.8% (found); R251 = 0. Paper chromatography: W1 23x62, eluent: n-BuOH-AcOHPyr.-water 30: 6: 24: 20. Yield in summary: 36.82% (from the protected monomer); 18.85% (from the

  free lysine). PM = 5.500 - 20.300 3.

  h) Purification of the crude end product 250 g of the polymer obtained in step g) was dissolved in water and then chromatographed on a Sephadex G-50 (fine) column. Eluent: distilled water, 0.9% NaCl solution or 0.1 N HCI solution. Detection for fractions (1 ml)

at 220 nm UV.

  i) Purification of the crude end product 2 g of the poly-e-L-lysine hydrobromide prepared in step g) was dissolved in 2 ml of water and then 20 ml of ethanol was added. The polymer was precipitated with 80 ml of diethyl ether. After 24 hours of rest at 5 C, it was filtered, washed and dried. Yield: 1.26 g (83%) (57.7% when calculated

  for the protected tripeptide of departure).

  The substance thus obtained is called SZTP-14; PM =

  12.700; [t] 200 = +32.4 (c = 2, water); polydispersion = 3.7.

  The following substances have been prepared in a similar way:

  SZTP-15 PM = 11.600 200 [] 20D = +31.9

  SZTP-16 MW = 13.400 200 [x] 20o = +32.6

  SZTP-17 MW = 14,500 200 [1] 20 = +32.1

EXAMPLE 2

  Poly-c-L-Lysine Hydrochloride A solution of 1 g of the polymer prepared according to the procedure of Example 1 in 40 ml of a 2M NaCl solution was dialyzed against a 2M NaCl solution and then water. After

Freeze-drying, 0.85 g of the polymer was collected.

EXAMPLE 3

  Poly-c-L-lysine (free base) An ion exchange column (40 μl) was prepared from a Dowex 1-OH anion exchange resin. The solution in 1 ml of water, of 1 g of the polymer prepared according to the procedure of Example 1, was chromatographed at a flow rate of 1 cm3 / min. The fractions were analyzed, collected and lyophilized. We isolated

0.65% of polymer.

EXAMPLE 4

  Poly-e-D-lysine hydrobromide The procedure of Example 1 was repeated from

  10 g of the p-nitrophenyl ester of N-α-benzyloxycarbonyl-

  (N-s - tert-butyloxycarbonyl) -D-lysine. 1.3 g of the polymer was obtained.

  MW = 86,900; [0] 25D = -47.08 (c = 0.92, water).

EXAMPLE 5

  Poly-6-L-Ornithine The procedure of Example I was repeated from 10

  p-nitrophenyl ester of N-α-benzyloxycarbonyl

  (N-6-tert-butyloxycarbonyl) -L-ornithine and obtained 7.9 g of

  N-α-benzyloxycarbonyl p-nitrophenyl ester hydrochloride

  L-ornithine. P .: 87-90 C. This salt has been coupled in the way

  described in step 1b) with N-α-benzyloxycarbonyl-

  (N-6-tert-butyloxycarbonyl) -L-ornithine and gave a dipeptide,

  N-α-Z-N-6- (N'-α-Z-N'-6-tert-butyloxy) p-nitrophenyl ester

  carbonyl-L-ornithyl) -L-ornithine (12.1 g); m.p.:133-136 C. By repeating step a1 from the dipeptide, the

2622 1 9 5

  N-α-Z-N-6- (N'-a-ZN'-6-) p-Nitophenyl ester hydrochloride

  L-ornithyl) -L-ornithine (11.99 g, mp 115-119 ° C), from which the p-nitrophenilic ester of N-6-Boe-tri (NcZL-ornithine) (14.8 g mp: 150-153 ° C) was obtained by coupling according to the method of step 1b). The p-nitrophenyl salt of tri (β-aZL-ornithine) was prepared according to the method of step 1c), mp: 115-130 ° C. 22 g of this product were polycondensed according to the procedure of step lf), which afforded 15.4 g of poly-1H-α-benzyloxycarbonyl-L-ornithine (87.6%). 13.1 g (93%) of hydrobromide was isolated from this product.

  poly-6-l-ornithine according to the method of step lg).

EXAMPLE 6

  Poly-6-D-Ornithine Hydrochloride The procedure of Example 5 was repeated from

  1 g of p-nitrophenyl ester of NLa-benzyloxycarbonyl-

  (N-6-tert-butyloxycarbonyl) -D-ornithine. 1.2 g (689%) of

poly-6-D-ornithine hydrobromide.

EXAMPLE 7

  Poly-β- (c, y-diaminobutyric acid) The procedure of Example 1 was repeated to obtain 6.9 g of p-nitrophenyl ester hydrochloride of α-benzyloxycarbonyl-L-α-acid. , y-diaminobutyric except for 9.8 g

  p-nitrophenyl ester of N-α-benzyloxycarbonyl acid

  H-γ-tert-butyloxycarbonyl-L-α, γ-diaminobutyric, m.p. 67-70 ° C.

  salt was coupled with N-α-benzyloxycarbonyl-N-y-ter-

  butyloxycarbonyl-L-x, y-diaminobutyric by the method of the step

  1b) to a dipeptide, the p-nitrophenyl ester of N-α-Z-N-acid

  / - (N'-a-Z-N'-tert-butyloxycarbonyl-c, y-diaminobutyryl) -c, y-di-

  aminobutyric (10.89, m.p .: 125-127 ° C). Repeating the procedure of Example 1a), 10.35 g of ester hydrochloride were synthesized.

  p-nitrophenyl acid N-α-Z-N-Y- (N'-α-Z-L-c, γ-diaminobutyryl) -

  Lu, γ-diaminobutyric acid from the dipeptide, mp: 108-112 ° C. From this salt, 13.7 g of p-nitrophenyl ester of Ny-BOC-tri (NaZL-acy-dianminobutyric acid was obtained. ) by coupling according to the procedure of step 1b), mp 143-145C. 10.5 g of p-nitrophenyl ester of tri- (NaZLx, ydiaminobutyric acid) were prepared from this product according to the method of step 1c), mp 105-110 ° C. This product was polycondensed in dimethylsulfoxide according to the procedure of the step lif) to give

  7.8 g (86.5%) of poly-Y-N-α-benzyloxycarbonyl-L-, γ-diamino-

  butyric. The protected polymer was treated according to the method described in step lg) to give 6.2 g (90.5%) of hydrobromide

  poly-y-L- (c, y-diaminobutyric acid).

EXAMPLE 8

  Poly-p-L- (ccp-diaminopropionic) Ride The procedure of Example 1 was repeated from

  9.6 g of p-nitrophenyl ester of N-α-benzyloxycarbonyl-N-8-

  tert-butyloxycarbonyl-L-op-diaminopropionic acid, to obtain 7.1 g of p-nitrophenyl ester hydrochloride of N-α-benzyloxycarbonyl-L-, p-diaminopropionic acid m.p .: 65-67 ° C. This salt

  was coupled with N-α-benzyloxycarbonyl-N-B-tert-butyloxy acid

  carbonyl-L-p-diaminopropionic acid by the method of step 1b) and

  gave 11.0 g of dipeptide, the p-nitrophenyl ester of N-acid

  α-Z-N- (N, N'-N, N'-S-butyloxycarbonyl);

  diaminopropionic, mp: 11: 9-122 ° C). Repeating the procedure of Example 1a), 10.85 g of ester hydrochloride were synthesized.

  p-nitrophenyl acid N-α-Z-N-e- (N '-α-Z-L-α, e-diaminopropionyl) -

  L-α-diaminopropionic from the dipeptide; yield: 10.85 g, mp: 10 2 -105 ° C. From this salt, 12.6 g of ester were obtained.

  N-α-BOC-tri- (N-α-Z-L-xβ-diamino) p-nitrophenyl acid

  propionic) by coupling according to the procedure of step 1b), mp: 138.degree.-140.degree. C. Prepared from this product,

  12.4-9 p-nitrophenyl ester of tri- (N-ca-Z-L-c, p-diamino)

  propionic) according to the method of step 1a), mp: 102-106 ° C. This product was polycondensed in dimethylsulfoxide according to

  procedure of step lf) to give 8.7? g (90.2%) of poly-p-acid

  e-a-benzyloxycarbonyl-L-a, Ii-diaminopropionic. The protected polymer

2622 1 95

  was treated according to the method described in step lg) to give 6 g (88.7%) of poly-p-L-o, p-diaminopropionic acid broahydrate.

EXAMPLE 9

  Injection solution 500 mg of poly-L-lysine, HBr are dissolved in 10 ml of physiological NaCl solution. This injection solution is filtered to be sterilized. The dose is 1-100 mg / kg / day,

depending on the condition of the patient.

EXAMPLE 10

  Preparation of tablets Tablets are prepared from the following components: poly-L-lysine, HCl lactose 16 g sucrose corn starch 2 g magnesium stearate 2 g alginic acid These materials are homogenized and 100 tablets are

pressed from the mixture.

EXAMPLE 11

  Method for increasing the resistance of plants to a

viral infection.

  An aqueous solution is prepared from 5 g of polyiso-L-lysine with 100 l of water. The pH of the solution is set at 7.0-7.2 with a solution of NaOH, and then 1 ml of Tween 20 is added. This solution is applied on a surface of 1 ha, on

  plants to protect while they are in post-emergence.

  REUENDICRTIOHS 1.Composition having the general formula (1)

6 1 8

R R R R

H HN-C! C C CO OH

17 I I9 M1

  Wherein R4, R5, R6, R7 and R10 independently of one another represent a hydrogen atom or a methyl group,

C1-4 alkyl.

  RB and R9 are each hydrogen, r is an integer of 10 to 400, a is 0, 1, 2 or 3, and

n is 0 -

  their salts, their racemates and their optical isomers.

  2. Compounds having the general formula (1) - wherein R4, R5, R6, R7 and R10 represent a hydrogen atom, r is a value of 10 to 400, m is 0, 1, 2 or 3 and is 0 - and their

  salts, their racemates and their optical isomers.

  3. Polyisolysin having an average molecular weight of

5.500 to 25.600.

  4. PoyiYsoornithine having an average molecular weight

from 4,400 to 13,500.

  5. Pharmaceutical preparations having an antitumor effect and an antiviral effect, comprising as active agent a compound of general formula (1) in which R4, R5, R6, R7, R3, R9 and R10 represent a hydrogen atom, r a value of 40 to, a is 0, 1, 2 or 3 and n is 0 and the amino groups have an L configuration, mixed with carriers and / or additives

  commonly used in the pharmaceutical industry.

2622 1 9 5

  Pharmaceutical preparations according to claim 5, wherein the active agent is a

  polyisolysin having an average MW of 5,500 to 22,500.

  A process for preparing polysiloxynocarboxylic acids having the general formula (1) - wherein R4, R5, R6, R7, R8, R9 and R10 are the same or different and may be hydrogen or C1-4alkyl, r is an integer of 10 and 400, * is an integer of 0 to 10, and

n is an integer from 0 to 10 -

  their salts, racemates and optical isomers, by coupling the corresponding monomeers and polycondensation oligomers obtained in this manner, characterized in that a monomer or an oligomer of general formula (11),

4 6H [0 0

R R

[I | I _ 1

H Hi'I-C C- Cc CO OR)

7 1, 1 9 (II)

  ## STR2 ## wherein q is an integer of 1 to 9, R 1 is an amino protecting group and all R 1 are the same in oligomers, R 3 is an active ester group protecting for carboxy groups, suitable for protection and simultaneous activation, R4, R5, R6, R7, R8, R9, R10, a and n are as defined above, and in the case of oligomers, the substituents may be identical or different - is coupled with a monomer of general formula (111)

2622 1 95

R2 H11 C- - C C O

RLHN CO / \

1 5 17 I

R PR NH

  Wherein R2 is an amino protecting group other than R1, which may be selectively removed from R1, X is a carboxy activating group provided that the -COX group is more active than the COOR3 group, R1 , R4, R5, R6, R7, R8, R9, R10, and n are as defined above - and from the compound of general formula (IVU) thus obtained

4-6 0 /

2 HN -C C C C - OR3

  Wherein the substituents are as defined above and s is an integer of 2 to 10 - the protecting group R2 is removed, then, if s <9, the resulting compound is treated with a monomer of the general formula (III) or it is polycondensed in a known manner and if desired, when the obtained compound of the general formula (1) is in the form of a free base, it is converted into a salt, or it is a salt, it is transformed into a free base or another salt. 8. The process of claim 7 for producing

  polyisodiaminocarboxylic acids of general formula (1) -

  wherein R4, R5, R6, R7, R10 and r are as defined in

2622 1 95

  Claim 1, a is 3 and n is O - and their salts, characterized

  in that a monomer or an oligomer of the general formula (11),

  wherein R1, R1, R5, R6, R7, R10 and q are as defined in claim 1, and a and n are as defined above - is treated with a monomer of general formula (111) - in which R1 , R2, R4, R5, R6, R7, R10 and X are as defined

  in claim 1 and at and n are as defined above -

  and from the compound of general formula (IU) thus obtained - wherein the substituents are as defined above and is as defined in claim 1 - the protecting group R2 is removed, and then, if desired, when 9, the compound obtained is treated with a monomer of general formula (III) or

is polycondensed in a known manner.

  9. The process of claim 7 for producing

  polyisodiaminocarboxylic acids of general formula (1) -

  in which R1, R3, R14, R5, R6, R7, R10 and q are as defined in claim 1, a and n are as defined above - and their salts, characterized in that a monomer or oligomer of general formula (11) - wherein R 1, R 3, R 4, R 5, R 6, R 7, R 10 and q are as defined in claim 1, and m and n are as defined above - is treated with a monomer of general formula (111) - wherein R1, R2, R4, R5, R6, R7, R10 and X are as defined in claim 1 and at and n are as defined above - and then from the compound of the general formula (1U) thus obtained - wherein the substituents are as defined above and is as defined in claim 1 - the protecting group R2 is removed and, if desired, when if 9, the compound obtained is treated with a compound of general formula (111) where it is polycondensed

known.

  10. Process according to claim 7 for preparing polyiso-llysines and their salts, characterized in that N-α-Z-L-Lysine pnitrophenyl ester hydrochloride and N-α-Z-N-e-BOC-L-lysine, converted into mixed anhydride with the

  isobutyl chloroformate, are used as starting materials.

2622 1 95

  11. Process according to Claim 7 for preparing polyiso-Lornithines and their salts, characterized in that Nitrophenilic acid ester hydrochloride of N-ZL-ornithine and Na-ZN-BOC-L-ornithine, converted into anhydride. mixed with isobutyl chloroformate, are used as starting materials. 12. Process according to Claim 7 for preparing polyiso-D-lysines and their salts, characterized in that the

  p-nitrophenyl ester hydrochloride of N-a-z-D-lysine and N-c-

  -Z-N-t -BDOC-D-lysine, converted into mixed anhydride with the

  isobutyl chloroformate, are used as starting materials.

  13. A process according to claim 7 for preparing polyiso-Dornithines and their salts, characterized in that N-a-Z-D-ornithine pnitrophenyl ester hydrochloride and N-α-Z-N-B-BOC-D-ornithine, converted into mixed anhydride with

  isobutyl chloroformate, are used as starting materials.