DE3835962A1 - POLYISODIAMINOCARBONIC ACIDS, METHOD FOR THE PRODUCTION THEREOF AND THE MEDICAMENTS AND PLANT PROTECTION AGENTS CONTAINING THEM - Google Patents

Abstract

The invention relates to isopolypeptides of the general formula (I> <IMAGE> as well as their salts, racemates and optical isomers, and to a process for preparing these compounds. The compounds exert an antitumour and antiviral effect. In the above formula R<4> to R<10> are each hydrogen or C1-4-alkyl, r is 10 to 400, m is 0 to 10 and n is 0 to 10. Compound of the above formula wherein m is 0 to 3, n is 0 and R<8> and R<9> are each hydrogen are defined per se.

Description

The invention relates to novel polyisodiaminocarboxylic acids, a process for their preparation and medicaments containing these compounds, in particular with antitumor and antiviral activity, and plant protection products, in particular for increasing the resistance of the plants against Virus infections.

In nature also come from Diaminomonocarbonsäuren built, containing isopeptide bonds peptides, for Example in some peptide antibiotics, in the cell wall of some bacteria as well as in the clavicepamines. possibly For example, the polypeptides may exert biological effects and also be produced organically.

It has been found by Japanese researchers that the microorganism Streptomyces albulus also produces an L-lysine polymer consisting of 25 to 30 monomer units, wherein the monomers are linked by isopeptide bonds as detected. (S. Shima and H. Sakai: Agric., Biol. Chem. 45, [1981], 2 503). It has been demonstrated that this natural ε- poly-L-lysine exerts a strong bacteriophage inactivating action (Shima S. and coworkers: Agric. Biol. Chem., 46, [1982], 1917) and a fermentation process for the preparation of this Isopeptides (S. Shima and coworkers: Nippon Nogeikaku Kaishi 57, [1983], 221).

From the human therapeutic point of view, the basic proteins that can be isolated from the saprophytic spice of the ergot [Claviceps purpurea - Fr. (Tul.)], The clavicepamine, are of importance. These proteins can be separated into fractions with molecular weights of 2,000 to 17,000, the lysine content of which is extremely high (30 to 90 mol%). Previous studies have confirmed that proteins inhibit cell proliferation and thereby the growth of animal tumors with virtually no toxic symptoms (E. Thihák: Dissertation for Candidacy, 1978). Structural analysis of the isolated lysine-rich proteins proved that the ε- amino groups of the L-lysine units are in peptide bond (Gy Szókán and co-workers: J. Chromatography 366, [1986], 283). However, based on the literature and own investigations, it is not known what role the polyisolysine structural units play in the beneficial biological effect of clavicepamine.

Indian researchers investigated the antiviral activity of synthetic poly-L-isolysines, their derivatives acylated with amino acids, and synthetic poly- β -L- α , β -diaminopropionic acid. It has been confirmed that these compounds have no immediate effect but induce an interferon-like antiviral factor but weaker than poly-L-lysine (Indian J. Exper. Biology 20, [1987], 227; Indian J. Chem. 1982], 483).

To investigate the relationships between structure and biological effect was the desire to be isopeptides of diaminocarboxylic acids with a number of elements from 10 to 400, with a given average molecular weight and with a low polydispersity in an also operational feasible way to produce. Basic Claims to the substances to be produced were: higher Purity, low scattering of the molecular weight in the Trap of a given average value (low polydispersity) and compliance with the pre-planned configuration the amino acid residues. It is well known that the biological effect of the biopolymers by their molecular weight distribution is strongly influenced. While own Previous studies have shown that the biological Effect of amino acids and polypeptides by their optical configuration is decisively influenced. The Molecules of opposite configurations may be one  have opposite effects; this was at several amino acids observed.

For example, the growth of vaccinable animal Tumors inhibited by the D-arginine monomer, by but stimulates the L-arginine monomer (B. Szende and E. Thyhák: Lancet 7 456 [1968], 824; B. Szende: Dissertation for the candidature, 1974). Accordingly, the D and L-amino acids cancel each other out of their effects when they in a racemic mixture or in a racemic polymer coexist.

For the preparation of the polymers with a number of ring members From 10 to 400 seemed a synthetic way, especially by the polymerization of monomers to be feasible.

The meaning of those used in the description Abbreviations are given in accordance with the IUPAC-IUB Comission on Biochemical Nomenclature (J. Biol. Chem. 247 [1972], 977) indicated below.

Z: benzyloxycarbonyl group
Boc: tert-butyloxycarbonyl group
Np: p-nitrophenyl group
Su: N-hydroxysuccinimido group
Pcp: pentachlorophenyl group
PfP: pentafluorophenyl group
Bt: N-hydroxybenzotriazolyl group
Bu ⁱ OCO: isobutyloxycarbonyl group
EtOCO: ethyloxycarbonyl group
N₃: azido group
EEDQ: N-Ethyloxycarbonyl-2-ethoxy-1,2-dihydroquinolinyl group
IIDQ: N-Isobutyloxycarbonyl-2-isobutyl-1,2-dihydroquinolinyl group
THF: tetrahydrofuran
DMF: dimethylformamide
HPLC: high performance liquid chromatography
OPLC: Overprint Thin Layer Chromatography
IEF: isoelectric focusing
MS: mass spectrometry
NMR: Magnetic Nuclear Magnetic Resonance Spectroscopy
Z _CO₂ : Z group, measured on the basis of carbon dioxide.

Using the method of Hull and coworkers (Biopolymers 17 [1978], 4227), an active ester (-OPcp) of monomeric lysine protected at the α- amino group by a Z group was polycondensed to isopolylysine. Of Gangopadhyay and Mathur (Indian J. Chem. 21B [1982] 483) was a poly (β-L-α, β-diamino-propionic acid) -isopeptid by polycondensation of Pentachlorphenylesters of the α-amino group at the a-Boc protecting group having produced lysines. Lysine protected by a Z group was polymerized by diphenylphosphoryl azide (DPPA) according to Nishi and coworkers (Int. J. Biol. Macromol., 2 [1980], 53).

It is noted that the authors of the above disclosures no information about the molecular weight, the molecular weight distribution and communicated the optical purity of their products had. Own experiences have shown that - in Comparison to your own goals - only polyisolysins with a low molecular weight (with a number of links from 15 to 20, that is with a molecular weight of at most 2 500). The paper chromatographic Testing the products showed that they are strongly polydispersed were and the yield considering the practical Viewpoints remained low. At own biological Investigations proved these products to be ineffective.  

For the preparation of polymers with a low polydispersity, the methods of Kushwaha et al (Biopolymers 19 [1980] 219) and those of Mathur and co-workers (Int. J. Peptide Protein Res. 17 [1981], 189) appeared to be most suitable his. In the first mentioned reference, the preparation of ε- polylysine and in the latter of the δ- polyornithine is described. The basic principle of both methods is as follows. A tripeptide methyl ester is prepared from a suitably protected amino acid methyl ester with a protected active ester or azide, and its carboxyl group is liberated by alkaline hydrolysis and converted to an activated derivative, for example a pentachlorophenyl ester. By removing the terminal ω -NH₂ protective group, an activated tripeptide suitable for polycondensation is obtained. After polycondensation, the α- amino groups are released and the isopropeptide is recovered. In said processes, a Boc-protecting group in the a- position and a Z-protecting group in the ω- position are used; the latter is removed by catalytic hydrogenation. In these publications, molecular weight, polydispersity, and optical purity (L and D configuration) of the final products were not characterized.

During reproduction, a significant disadvantage of the latter two methods was noted. This is because in the course of the preparation of the oligomer, the carboxyl group not participating in the reaction is protected in the form of its methyl ester, and therefore, the oligomer thus obtained is unsuitable for direct polycondensation. To form the activated derivative, the carboxyl group is released by alkaline hydrolysis, and this step is likely to result in disadvantageous racemization, reduction in yield, and undesirable side reactions. At the same time, the polymers thus obtained have a strong polydispersity. Furthermore, the use of the Z-protecting group in the ω- position and the removal of this group by catalytic hydrogenation are disadvantageous.

The polyisolysines obtained by the latter methods proved in own biological investigations as ineffective.

The invention is therefore based on the object, under Overcoming the disadvantages of the prior art new iso-polypeptides, which has a certain molecular weight and a have low polydispersity and diaminocarboxylic acids built with a specific (D or L) configuration are, with superior pharmacological, especially tumor-inhibiting and antiviral, effects as well as a procedure for their preparation and containing these compounds To create medicines.

The above was surprisingly by the invention reached.

The invention is based on the surprising finding that the above can be achieved when first for polycondensation directly suitable, from Diaminomonocarbonsäuren built-up activated monomers or oligomers in such a way be prepared that the carboxyl group of the starting materials protected by an active ester group and the Peptide coupling by means of a process under its conditions the one acid amide bond forming at the carboxyl group activated derivative with the amine component with a much higher speed than the speed the aminolysis of the active ester protecting group becomes.  

Furthermore, the invention is based on the surprising Finding that in the case of diaminomonocarboxylic acids the p-Nitrophenylestergruppe for temporary protection and to simultaneous activation of the carboxyl group and the process the mixed anhydrides to form the peptide bond are very suitable and appropriate to the acid amide bond participating amino group through a Boc group and the amino group not participating in the reaction by a Z group are protected.

Finally, the invention is based on the surprising Finding that the homo-peptides of the sequence isopeptides, that is, consisting of identical and homologous amino acids Isopeptides of any diaminocarboxylic acid prepared by the above method in the manner can be that the amino acid residues their original Retain (L. or D) configuration.

The invention relates to polyisodiamine carboxylic acids the general formula

wherein
R₄, R₅, R₆, R₇, R₈, R₉ and R₁₀ independently of one another represent hydrogen or alkyl radicals having 1 to 4 carbon atoms,
r is an average of 10 to 400,
m is a number from 0 to 10 and
n represents a number from 0 to 10,
and their salts, optical isomers and racemates.

The meanings of the corresponding substituents in the individual So members can be the same or different.

Preferably m is a number from 0 to 3, in particular 2 or 3, and / or means
n 0.

It is also preferred that the or the alkyl radical (s), for which [n] R₄, R₅, R₆, R₇, R₈, R₉, and / or R₁₀ may or may be, [a] such [r] with 1 or 2, in particular 1 carbon atom (s) is or are.  

Further, it is preferable that r is a number of 40 to 200.

It is also preferred that the invention Polyisodiaminocarbonsäuren have the L-configuration.

Furthermore, it is preferred that R₈ and R₉ are hydrogen atoms mean. Particularly preferred are beyond also R₄, R₅, R₆, R₇ and R₁₀ for hydrogen atoms.

A preferred group of Polyisodiaminocarbonsäuren invention are those in which R₄, R₅, R₆, R₇ and R₁₀ independently of one another hydrogen or alkyl radicals having 1 to 4 carbon atom (s), R₈ and R₉ are hydrogen, r is a number from 40 to 200 , m is 3 and n is 0. Of these substances, those are particularly preferred in which R₄, R₅, R₆, R₇ and R₁₀ are hydrogen and r is a number from 80 to 120.

Another preferred group of Polyisodiaminocarbonsäuren invention are those in which R₄, R₅, R₆, R₇ and R₁₀ independently of one another hydrogen or alkyl radicals having 1 to 4 carbon atom (s), R₈ and R₉ are hydrogen atoms, r is a number from 10 to 120 means m 2 and n is 0.

A preferred group of Polyisodiaminocarbonsäuren invention are those in which R₄, R₅, R₆, R₇ and R₁₀ independently of one another hydrogen or alkyl radicals having 1 to 4 carbon atom (s) Polyisodiaminsäuren, R₈ and R₉ are hydrogen, r is a number from 10 to 400 means m represents 1 and n is 0.

Yet another preferred group of Polyisodiaminocarbonsäuren invention are those in which R₄, R₅ and R₁₀ independently of one another hydrogen or alkyl radicals having 1 to 4 carbon atom (s). R₆, R₇, R₈ and R₉ are each hydrogen, r is a number from 10 to 400 and m and n are 0.

The salts of the polyisodiaminocarboxylic acids according to the invention are useful those with pharmaceutically acceptable acids. Examples are acid addition salts, such as hydrochlorides, hydrobromides, Hydroiodides, sulphates, phosphates, nitrates, oxalates, Fumarates, gluconates, tartrates, maleates, acetates, citrates, Benzoates, ascorbates, lactates and alginates.

Particularly preferred polyisodiaminocarboxylic acids according to the invention are polyisolysin with an average Molecular weight of 5,500 to 25,600 daltons, in particular 5 500 to 22 500 daltons, especially 10 200 to 15 400 Dalton, and polyisoornithine with an average Molecular weight of 4,400 to 13,500 daltons.

The invention also provides a process for the preparation the compounds of the invention, which thereby characterized in that protected monomers or oligomers with [a] diamino acid unit (s) of the general formula

wherein
q is a number from 1 to 9,
R₁ is an amino protecting group and is the same in oligomers in each oligomeric unit,
-O-R₃ is an active ester protecting group which serves to protect and simultaneously activate the carboxyl group,
R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, m and n have the meanings given above,
wherein in the case of oligomers, the meanings of the substituents in the individual members are the same or different, with protected monomeric diamino acid derivatives of the general formula

wherein
R₂ is an amino-protecting group different from R₁ and selectively removable from R₁,
X is a group used to activate the carboxyl group, with the proviso that the group of the general formula

responsive as the group of general formula

is, means and
R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀ m and n have the meanings given above,
be reacted, from the resulting protected oligo- or polyisodiaminocarboxylic acids of the general formula

wherein
R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀ m and n have the meanings given above and
s is an integer from 2 to 10,
the amino protecting group R₂ is split off and from the obtained protected oligo- or polyisodiaminocarboxylic acids, in the case that s 2 to 9, after their reaction with protected monomeric diamino acid derivatives of the general formula III and / or polycondensation, the protecting groups R₁ are eliminated, which is known per se If appropriate, the resulting Polyisodiaminocarbonsäuren the general formula I are converted into acid addition salts or optionally the resulting acid addition salts of Polyisodiaminocarbonsäuren the general formula I are transferred to the free Polyisodiaminocarbonsäuren or other acid addition salts and / or optionally cleavage of the resulting racemic Polyisodiaminocarbonsäuren the general formula I or salts the same is carried out in their optically active antipodes or a racemization of the corresponding optically active compounds.

Preferably, they are of the type used in peptide chemistry known, divergent, selectively removable Amino protecting groups R₁ and R₂ R₁ represents a Z group and R₂ is a Boc group.

It is preferred as the active ester protecting group for which -O- R₃, a group -ONp, -OSu, -OPcp, -OPfp or -OBt to use.

Suitably, as the group serving for activating the carboxyl group, X represents a group Bu ⁱ OCO-, EtOCO-, N₃-, -OSu, OPcp or -OPfp depending on the reaction conditions and the meaning of R₃ used. Preferred meanings of X are the groups Bu ⁱ OCO- and EtOCO.

According to an expedient embodiment of the process according to the invention for the preparation of polyisodiaminocarboxylic acids of the general formula I,
at which
m 3 means and
n is 0,
are used as protected monomers of the oligomers with [a] diamino acid unit (s) of the general formula II those
at which
m 3 means and
n is 0,
used as protected monomeric diamino acid derivatives of the general formula III those in which
m 3 means and
n is 0,
used.

According to a further expedient embodiment of the process according to the invention for the preparation of polyisodiaminocarboxylic acids of the general formula I,
at which
m 2 means and
n is 0,
are used as protected monomers or oligomers with [a] diamino acid unit (s) of the general formula II
at which
m 2 means and
n is 0,
used as protected monomeric diamino acid derivatives of general formula III, those
at which
m 2 means and
n is 0,
used.

According to the most preferred embodiment of the process according to the invention are used as starting materials monomers of the general formula III in which R₁ is a Z group, R₂ is Boc and -O-R₃ is a -ONp group used. There are the Boc-protecting group in a conventional manner, preferably by acidolysis, cleaved and then coupled as protecting groups R₁ = Z and R₂ = Boc having Diaminosäurederivate the general formula III according to the procedure of the mixed anhydrides. The dipeptides thus obtained can be polycondensed after removal of the Boc protecting group to the Polyisodiaminocarbonsäuren with the desired number of members or preferably by coupling a further protected amino acid at most Dekapeptide, most preferably converted into tripeptides, and then the resulting oligomers expediently by polycondensation in the Polyisodiaminocarbonsäuren be transferred with the desired number of limbs. From the latter, the α -Z protective groups are split off with hydrogen bromide in glacial acetic acid and the end product is isolated.

The optical isomers of the polyisodiaminocarboxylic acids become expediently manufactured in such a way that the corresponding Isomers are used as starting materials. When racemic Polyisodiaminocarbonsäuren are produce expedient the racemates of the monomers as starting materials used.  

According to a specific preferred embodiment of the process according to the invention, a protected monomer with [a] diamino acid unit (s) of general formula II is N- ^α- ZL-lysine p-nitrophenyl ester hydrochloride and protected monomeric diamino acid derivative of general formula III is a mixed anhydride which is selected from N ^α- ZN ^ε -Boc-L-lysine with isobutyl chloroformate.

According to another particular preferred embodiment of the process according to the invention, the protected monomer with [a] diamino acid unit (s) of general formula II is N- ^α- ZL-ornithine p-nitrophenyl ester hydrochloride and protected monomeric diamino acid derivative of general formula III is a mixed anhydride, which has been obtained from N ^α -ZN ^β -Boc-L-ornithine with isobutyl chloroformate.

According to a further specific preferred embodiment of the process according to the invention, the protected monomer with [a] diamino acid unit (s) of general formula II is N- ^α- ZD-lysine p-nitrophenyl ester hydrochloride and protected monomeric diamino acid derivative of general formula III is a mixed anhydride, which has been obtained from N ^α -ZN ^ε -Boc-D-lysine with isobutyl chloroformate.

According to yet another specific preferred embodiment of the process according to the invention is a protected monomer with [a] diamino acid unit (s) of general formula II N ^α -ZD-ornithine p-nitrophenylesterhydrochlorid and protected monomeric diamino acid derivative of general formula III, a mixed anhydride, which from N ^α -ZN ^β -Boc-D-ornithine with isobutyl chloroformate.

Through the systematic investigation of itself as am Most preferred coupling procedure that That is, the procedure of mixed anhydrides were the following reaction conditions as most suitable detected:

• a) solvent: acetonitrile, THF, DMF or Dichloromethane.
• b) anhydride-forming reagent: ethyl or Isobutyl chloroformate, EEDQ, IIDQ in the presence a tertiary base, for example of triethylamine, diisopropylethylamine or N-methylmorpholine.
• c) Activation time and temperature: 10 to 30 minutes at temperatures from -10 ° C to -20 ° C.
• d) release of the amine component from its Salt with triethylamine or diisopropylamine in the reaction mixture.

In this way, the pure end products were in average yields of 60 to 80%.

With the method according to the invention can from diaminocarboxylic acids constructed with both L and D configurations Homo and sequence polyisodiaminocarboxylic acids [homo- and sequence isopeptides] while retaining the original one Configuration of the starting materials are produced.

For the analysis of the produced polyisodiaminocarboxylic acids became modern separation techniques (for example HPLC, OPLC, IEF) and structure elucidation procedures (for example MS, NMR). Through this Investigation procedures became the success of the production promoted the desired substances.  

The biological studies were conducted in the in the Examples described using purified products. If necessary, the cleaning was done by ordinary other operations, for example column chromatography and gel chromatography, supplemented.

Compared to those of Kushwaha or Mathur and of Hull communicated to the above mentioned authorities Syntheses are the advantages of the method according to the invention summarized as follows:

• A) It can be products with a favorable biological Effect made reproducible become.
• B) A directly suitable for polycondensation Oligomer is obtained in such a way that mentioned above in the known methods Step of hydrolysis is avoided, causing Racemization is avoided, which means the biological effect is crucial.
• C) Products with certain average molecular weights within wide limits, whose molecular weight distribution is surprisingly favorable (less polydisperse), can be produced, which is also advantageous for achieving the desired biological effect. Accordingly, the data of the molecular weight measured by ultracentrifugation of the substance of the invention polyiso-L-lysine hydrobromide [SZTP-14] are as follows: molecular weight: 12,700 ± 200; Polydispersity, that is
• D) It is achieved a higher yield and the optical purity of intermediates and final products is higher. Both the intermediates as well as the final products are well crystallizable and therefore easily isolable substances.
• E) The individual steps of the peptide coupling go during a much shorter time.
• F) By the amino-protecting group combination, the in the embodiment identified as most favorable used the method according to the invention can be, by catalytic hydrogenation performed release of the acid amide bond participating amino group can be avoided. That's in terms of a technical or industrial implementation favorable.
• G) A significant advantage of the invention Method is that if of optical pure isomers, the polymer won with the appropriate configuration becomes. This advantage arises from the cheap selected steps of the synthesis.

As mentioned above, using the above-described received known methods ineffective products.  This is above all the polydispersity of the final product as well - in the case of Kushwaha or Mathur, respectively - the racemization of the final product used in the synthesis of occurs for the polycondensation suitable oligomer, attributed. In contrast, according to the invention Substances with surprisingly favorable biological properties obtained, as illustrated in more detail below becomes.

The invention also relates to medicaments which by a content of 1 or more compound (s) of the invention as active ingredient (s), optionally together with 1 or more common pharmaceutical carrier and / or excipient (s), Marked are.

The polyisodiaminocarboxylic acids according to the invention have namely as already said valuable pharmacological, in particular antitumor and antiviral effects, such as Resistance to virus infections.

These are particularly pronounced in those with L configuration.

The invention also pesticides, especially for increasing the resistance of plants against Viral infections, which by a content of 1 or more Compounds of the invention, optionally together with 1 or more carrier and / or vehicle carriers usual in pesticides or excipient (s).

The polyisodiaminocarboxylic acids according to the invention have namely also valuable pesticide effects, in particular antiviral effects.

The biological activity of the compounds according to the invention was tested under in vitro and in vivo conditions.  

The results are summarized below. The tumor-inhibiting Effect was on animal tumor strains, the antiviral Effect using a plant virus educated.

In the experiments, the polyiso-L-lysines proved with a number of limbs from 40 to 200 (with a molecular weight from 5,500 to 25,600) as particularly effective; a great one The products had a good number of effects from 80 to 120 (with a molecular weight between 10,200 and 15,400).

Testing the tumor-inhibiting effect

Testing the effect of affecting cell proliferation Compounds generally occurred in vivo using cell cultures and in vivo using inoculable animal tumors. That's why the test substances are involved in a human erythroleukemia Cell culture (K-562) (origin: Karolinska Institute, Stockholm) and at four durchfpfbaren mouse tumors (L₁₂₁₀ lymphoid Leukemia, P₃₈₈ Tumor macrophage origin, Honest Carcinoma and Lewis lung carcinoma). In case of L₁₂₁₀, P₃₈₈ and Ehrlich tumors served the survival of Animals as parameters, while in the case of Lewis lung carcinoma the number and size of the tumor induced Lung and liver metastases in the treated animals and in the untreated control animals. For these experiments, an injection solution was used, the 50 mg of test substance in 10 ml of physiological saline contained and sterile filtered.

The test substances were labeled as follows:  

Results of investigation of tumor cell proliferation

It was confirmed that the number of tumor cells in vitro under the effect of treatment with the compounds of the present invention decreased dose-dependently as compared with the control. This proved to be a stagnation in the number of cells in the case of smaller doses and in the destruction of the cells in the case of higher doses. Substances K-17 and H-17 were ineffective, and α- polylysine (Substance A-3) caused only a stagnation in the number of cells.

Due to the in vivo investigations revealed the treatment with SZTP substances the extension of the life span the treated animals in the case of tumors L₁₂₁₀ and  P₃₈₈. In the case of the Ehrlich tumor became a very long achieved tumor-free period through the treatment and the Tumor growth was only after interruption of treatment observed.

The metastasis of the Lewis lung tumor was influenced by the SZTP substances in such a way that the Formation of liver metastases after one in the spleen vaccinated primary tumor completely inhibited and the formation of pulmonary metastatic after vaccinated in the muscle primary tumor were significantly reduced.

Accordingly, the inventive proved Polyisolysins and polyisoornithines as tumor cell proliferation inhibiting Compounds under which polyisolysines a have particularly favorable effect. Regarding the Surviving the animals is having their humor-inhibiting effect the effect of the best known cytostatics comparable; It is particularly noteworthy that they are unexpected favorable inhibitory effect on the formation of metastases exercise. Another advantage of these compounds in comparison to the known cytostatics is that their toxicity is much lower than that of the present used cytostatics.

The biological tests have revealed that the therapeutic index of SZTP compounds, d. H. the quotient LD₅₀ / minimum effective dose above 10 while this value is common in most known cytostatics is under 10.

Regarding the biological effects showed There is no difference between male and female Animals, which is also considered cheap.

The tumor cell proliferation-inhibiting effect is illustrated by the following.

In vitro tests

Tables 1a, 1b and 1c show the effect of treatment with the polyiso-L-lysines prepared by various methods, with the polyiso-L-ornithine according to the invention and with the α- polylysine on the proliferation of the K-562 cells.

The parameters of these tests were as follows:
Cells: The K-562 cell line of human erythroleukemia (origin: Karolinska Institut, Stockholm) was used.
Number of cells: 5 × 10⁴ / ml; 5 ml in each test tube.
Number of samples: 3 test tubes for each dose.
Medium: M-199 medium (Parker medium) with 10% by weight calf whey (Flow Laboratories, Irvine, Scotland).
Temperature and atmosphere: 37 ° C; Mixture of 95% by volume of air and 5% by volume of carbon dioxide.
Treatment: 24 hours after dilution of the cultures.
Evaluation: Counting of the cells 24, 48, 72 and 96 hours after dilution in a Buerker chamber.
Number of trials: 2 with each substance.
Dose: 1-10-100 μg / ml

From Table 1a it can be seen that a significant dose-dependent proliferation inhibition occurred. It is noteworthy that a proliferation inhibition already in one Concentration of 1 μg / ml could be observed.

Table 1b showed that - in contrast to the above - the proliferation of K-562 cells by the substances H-17 and K-17 were not inhibited.

It can be seen from Table 1c that α- polylysine stagnated the number of cells compared to the control, but did not cause a reduction in the number of cells even at a higher concentration (100 μg / ml).

The values shown in the tables indicate the in the individual times observed number of cells. The Cells were used in the case of all three doses, respectively Control counted in the same time in each case 3 glasses. In the tables are these values and the deviation values summarized. The number of cells refers to Milliliter.  

Table 1a

Effect of the substances according to the invention (SZTP-14, SZTP-15, SZTP-16, SZTP-17 and SZTP-18) on the K-562 cell line in vitro (treatment: at 1, 10 and 100 μg / ml 24 hours after dilution the culture)

Complete disintegration was reported after a dose of 100 μg / ml, indicating an effective dose-dependent Effect indicates.  

Table 1b

Effect of comparison substances H-17 and K-17 on the proliferation of K-562 cells in vitro (treatment: at 1, 10 and 100 μg / ml 24 hours after dilution of the culture)

Table 1c

Effect of comparison substance A-3 on the proliferation of K-562 cells in vitro (treatment: at 1, 10 and 100 μg / ml 24 hours after dilution of the culture)

In vivo tests

a) L₁₂₁₀ tumor; Testing the substance SZTP-14.
Origin of the tumor: Chester Beatty Institute, London.
Animal strain: own DBA / 2 inbred mouse.
Weight and sex of the animals: 20-23 g; 5 females in each group,
Number of tumor cells: 10⁵ / animal, intraperitoneally (ip).
Treatment: 50 mg / kg ip starting at the 24th hour after the tumor is inoculated, once daily, for 8 days,
Control group: 0.2 ml physiological saline, ip
Evaluation: by the number of dead animals.
Results: Table 2 shows that the treated animals survived the control animals. While 10 days after vaccination of the tumor no control animal lived, all treated animals were still alive. The first treated animal died 13 days, the last 15 days after vaccination of the tumor.

Table 2

Survival of L₁₂₁₀ tumor vaccinated, treated with 50 mg / kg SZTP-14 DBA / 2 mice or DBA / 2 control mice

b) Testing the substance SZTP-14 on the ascites form of the P-388 tumor.
Animal strain: BDF / 1 hybrid, own inbreeding.
Weight and sex of the animals: males with 20-23 g body weight; 5 animals in each group.
Number of tumor cells: 2.8 × 10⁶ / animal, ip
Treatment: 10 mg / kg ip starting from the 24th hour after inoculation of the tumor, once a day, for 8 days. The control animals received at the same time 0.2 ml of physiological saline, ip
Evaluation: By the number of dead animals
Results: Table 3 shows the number of surviving animals. It can be confirmed that all the treated animals survived the control animals.

Table 3

Survival of P-388 tumor vaccinated, 10 mg / kg SZTP-14 treated BDF / 1 mice and BDF / 1 control mice, respectively

c) Examination of the substance SZTP-14 on Ehrlich ascites tumor. Animal strain: Swiss "non-inbred", own breeding.
Weight and sex of animals: 20 g body weight, 5 males and 5 females in each group.
Number of tumor cells: 10⁶ / animal, ip
Treatment: 50 mg / kg or 75 mg / kg ip starting 24 hours after tumor inoculation, once daily, for 20 days. The control animals received 0.2 ml physiological saline ip

table 4

Body weight shaping by Ehrlich ascites Tumor vaccinated, with 50 mg / kg and 75 mg / kg SZTP-14 i.p. Treated for 20 days, male and female Swiss Mice or Swiss control mice on the 16th day after vaccination beginning of the tumor; average initial body weight: 20 g. O: died as a result of the tumor; +: as a result other causes died.

Continuation of Table 4

Evaluation:
by constantly measuring the body weight and observing the transmissions. On the 55th day after the vaccination, the live treated animals were killed, dissected, the amount of ascites was determined and the eventual formation of a solid tumor was recorded.
Results:
Table 4 shows the body weight change of the mice from the 16th day after inoculation of the tumor. It is amazing that all control mice died between the 17th and 22nd day; The large excess of weight results from the high amount of tumorous ascites. In all groups of the treated animals, the animals - with the exception of one mouse in each group - still lived on the 55th day after vaccination of the tumor. At this point, the mice were killed. The amount of intra-abdominal ascites and the presence of a solid tumor are shown in Table 5. From this it can be seen that no animal was tumor-free at the moment of sacrifice, but 16 animals out of 20 treated animals still lived on the 33rd day after the last control mouse had died.

table 5

Ascites and solid tumor in the abdominal cavity of having Ehrlich Ascites tumor cells vaccinated with 50 mg / kg and 75 mg / kg, respectively SZTP-14 treated, killed on the 55th day of the trial Swiss mice.

treatment

Inhibition of tumor metastases by the substance SZTP-14

a) Lewis lung tumor (LLT).
Phylum. LATI C₅₇Bl mouse inbreeding.
Weight and sex of the animals: 4 treated and 6 control animals with 20-23 g body weight (females).
Number of tumor cells: 5 × 10⁶ LLT cells ip inoculated into the spleen.
Treatment: 75 mg / kg for 8 days after vaccination of the tumor.
Evaluation: by the number of liver metastases after killing the animals.
Results: the number of metastases is given in Table 6. It can be seen that no tumor formed after treatment with SZTP-14.

table 6

Number of liver metastases 8 days after vaccination LLT cells in the spleen of female C₅₇-Bl mice and Control mice (treatment daily with 75 mg / kg SZTP-14 i. p.)

b) Lewis lung tumor with intramuscular vaccination, lung metastasis count,
Animal strain: 5 female C₅₇Bl mice in each group.
Number of tumor cells: 5 × 10⁵ / animal intramuscularly, in the muscles of the right thigh. After 10 days, the tumorous extremity was amputated and the treatment was carried out from the 11th to the 17th day daily at 50 mg / kg ip. The control animals received 0.2 ml physiological saline ip
Evaluation: The animals were killed on the 18th day and the number of metastases and the average volume of metastases determined under a stereomicroscope.

table 7

Number of lung metastases 18 days after vaccination LLT cells and 8 days after removal of the primary tumor in C₅₇-Bl mice and control mice (treatment: daily with 50 mg / kg SZTP-14 i.p. between the 11th and 17th days)

Demonstration of the antiviral effect Inhibition of the synthesis of tobacco mosaic virus

Under normal greenhouse conditions, intact, sensitive to the infection by the tobacco mosaic virus Tobacco plants (Nicotiana tabacum cv. Samsun) mechanically inoculated (infected). The carborundum Powder with a particle size of 500 mesh used. The inoculum contained 200 μg / ml tobacco mosaic virus (U1 strain) in a 0.1 M phosphate buffer (Sörensen phosphate buffer, pH 7.2). The intact leaves were 3 hours after infection treated by spraying. The treatments were daily repeated once on three consecutive days. 2 weeks after the inoculation was the intensity of the symptoms the systematic infection was assessed and samples were taken from all leaf levels of the plant, d. H. from those in different Heights standing, taken, the accordingly different ages were. The virus content was determined from 1 g of frozen sample. The leaf samples were homogenized in a mortar with 4 ml Sörensen buffer (pH 7.2), then in Eppendorf tubes for 5 minutes with a  Centrifuged at 10,000 rpm. To the quantitative Virus determination was 15 ml of the supernatant (Supernatant) used. The virus content was determined by "rocket" immunoelectrophoresis by means of a virus-specific antigen (anti-TMV U1; 1 μl IgG / cm 2). The development agent was a 0.2 M sodium barbiturate-barbituric acid buffer solution. The virus concentration was reduced due to the standard Calibration curve from the height of the precipitation arc lines calculated.

Determination of virus content Treatment: with SZIP-14 virus content 200 mg / l 11 mg / ml 20 mg / l 24.66 mg / ml 2 mg / l 29.30 mg / ml 0 (control) 51.33 mg / ml

The polyisodiaminocarboxylic acids according to the invention can as active pharmaceutical ingredients both in the form of the free bases as well as in the form of their non-toxic pharmaceutically acceptable Salts are present as drug preparations. these can in a conventional manner by mixing with the at Pharmaceutical preparation customary carriers and / or excipients to be obtained.

Polyisodiaminocarbonsäuren invention as active ingredients containing drug preparations, for example Injection solutions, tablets, dragees, suppositories, Suspensions, drink solutions and powder mixtures.

The amount effective as a medicament active ingredient of the invention  Polyisodiaminocarboxylic acids depends on the route of administration and the condition of the patient. In general the daily dose is 0.02 to 20 mg / kg body weight, which is administered in 2 to 4 divided doses. In case of oral administration, higher doses are used.

Preferred pharmaceutical forms are the injection solutions, by intravenous, subcutaneous or intramuscular route be administered. For the preparation of injection solutions Be an isotonic saline solution, if necessary Phosphate buffer and ascorbic acid as stabilizer second hand.

Further examples of pharmaceutical forms are those for oral administration in solid or liquid form. in the Traps of liquid forms may include sweeteners and / or Taste Correctives as part of the pharmaceutically acceptable Diluent present. Advantageous fixed dosage units for oral administration are, for example, tablets, Dragees and capsules. For the production of these forms will or will the active substance (s) with the usual pharmaceutical or Carrier and / or excipient (s) mixed and the Mixture is pressed into tablets, converted to dragees or filled in capsules. As carriers may be conventional Filling and / or auxiliary materials, such as lactose, magnesium stearate, Sucrose, talc, stearic acid, gelatin, agar, Pectin, tragacanth, colloidal silica, corn starch and / or Alginic serve. The preparation of the suppositories serves preferably cocoa butter.

Furthermore, the polyisodiaminocarboxylic acids according to the invention both in the form of free bases and in form their salts are present in crop protection preparations.  

These can be mixed in a manner known per se with the usual in plant protection carriers and excipients to be obtained. These serve primarily the Increase the resistance of the plants against viral infections.

The solid or liquid preparations for the treatment of Plants are expediently prepared in such a way that the Polyisodiaminocarbonsäuren invention with conventional Carriers and / or additives, for example kaolin and / or Wetting agents, advantageously combined with other active ingredients, be mixed. From these preparations can solutions or suspensions containing the polyisodiaminocarboxylic acids according to the invention in concentrations from 0.0001 to 0.005% by weight [1 to 50 ppm], preferably 0.0005 to 0.0015 wt% [5 to 15 ppm], to be prepared. These can be in usual way, expedient in application rates of 0.5 to 5 g / ha.

The invention will be apparent from the following examples explained in more detail.

Example 1 ε- poly-L-lysine hydrobromide a) N ^α -benzyloxycarbonyl-L-lysine p-nitrophenyl ester hydrochloride

10 g of N ^α -benzyloxycarbonyl- (N ^ε- tert-butyloxycarbonyl) -L-lysine p-nitrophenyl ester are dissolved in 40 ml of trifluoroacetic acid and an equivalent amount of 2N hydrochloric acid ethyl acetate solution is added. After standing for 1 hour, the solution is evaporated to half and ether is added. The product is obtained with a yield of 8.3 g (96.06%), mp: 83-86 ° C, R _f = 0.68 (n-butanol-glacial acetic acid / water 4: 1: 1 v / v). Vol.).

b) N ^α -benzyloxycarbonyl-N ^ε - (N ' ^α -benzyloxycarbonyl-N' ^ε- tert-butyloxycarbonyl-L-lysyl) -L-lysine p-nitrophenyl ester

7.69 g (20 mmol) of N ^α -benzyloxycarbonyl- (N ^ε- tert-butyloxycarbonyl) -L-lysine are dissolved in abs. Dissolved acetonitrile and an equivalent amount of triethylamine (2.0 ml) and then isobutyl chloroformate (2.8 ml) are added dropwise. After activation, 9 g of salt prepared according to a) and simultaneously a solution of an equivalent amount of triethylamine (2.8 ml) in acetonitrile are added. The product is recovered by pouring the reaction mixture in water and recrystallized from the mixture of ethyl acetate and petroleum ether. Thus, 12.9 g (89.02%) of product are obtained, m.p .: 128-131 ° C. After analysis, it contains 99.8% -onp.
R _f = 0.64 (ethyl acetate / cyclohexane 3: 1 v / v)
R _f = 0.91 (n-butanol / glacial acetic acid / water 4: 1: 1 v / v)
HPLC: t _R = 8.8 minutes (k '= 2.2)
Column: ODS-Hypersil-6 (125 × 4 mm)
Eluent: methanol / acetonitrile / 0.02 M sodium acetate, pH 4.0, 40: 30: 30 v / v.
Speed: 1 ml / minute
Detection: by UV at 254 nm

c) N ^α -benzyloxycarbonyl-N ^ε - (N ' ^α -benzyloxycarbonyl-L-lysyl) -L-lysine p-nitrophenyl ester hydrochloride

A salt is prepared from 10.5 g of N ^α -benzyloxycarbonyl-N ^ε - (N ' ^α -benzyloxycarbonyl-N' ^ε- tert-butyloxycarbonyl-L-lysyl) -L-lysine p-nitrophenyl ester according to a). The yield is 10.25 g (98%), mp 109-113 ° C, R _f = 0.70 (n-butanol / glacial acetic acid / water 4: 1: 1 Vol./Vol.).

d) N ^ε- tert-butyloxycarbonyl-tri (N ^α -benzyloxycarbonyl-L-lysine) p-nitrophenyl ester

A mixed anhydride is prepared from 5.71 g (15 mmol) of N ^α -benzyloxycarbonyl- (N ^ε- tert-butyloxycarbonyl) -L-lysine with the equivalent amount of triethylamine and with isobutyl chloroformate in acetonitrile according to b). To this solution is added 10.5 g of salt of N ^α -benzyloxycarbonyl-N ^ε - (N ' ^α -benzyloxycarbonyl-L-lysyl) -L-lysine p-nitrophenyl ester in the presence of triethylamine. After working up according to b) 13.9 g of product are obtained, which is recrystallized from the mixture of acetonitrile and ether. This product melts at 145-150 ° C and, after analysis, contains 99.6% Sonp;
R _f = 0.98 (n-butanol / glacial acetic acid / water 4: 1: 1 v / v);
HPLC: t _R = 18.2 minutes, k '= 8.1;
Column: ODS-Hypersil-6 (125 x 4 mm);
Eluent: methanol / acetonitrile / 0.02M sodium acetate buffer (pH 4) 40: 30: 30 v / v; 1 ml / minute;
Detection: by UV at 254 nm.

e) tri- (N ^α -benzyloxycarbonyl-L-lysine) p-nitrophenyl ester hydrochloride

12.2 g of ester [prepared according to d)] are treated according to a). Thus, 12.1 g (95.4%) of salt are obtained, m.p .: 108-125 ° C, R _f = 0.71 (n-butanol / acetic acid / water 4: 1: 1 v / v).

f) ε- poly- [N ^α -benzyloxycarbonyl-L-lysine)

23 g (24 mmol) of tri (N ^α -benzyloxycarbonyl-L-lysine) - p-nitrophenyl ester hydrochloride are in 60 ml abs. Dimethyl sulfoxide, polycondensed in the presence of 4.5 ml of triethylamine until complete gelation. The polymer is isolated by pouring into sodium carbonate solution. Thus, 16.8 g (89.6%) of polymer is obtained. Analysis: Z _CO₂ calculated: 16.3%; found: 16.8%; [ η _sp ] / c = 31.92 ml / g 2 (c = 4.95 g / l, dichloroacetic acid).

g) ε- poly-L-lysine hydrobromide [polyiso-L-lysine hydrochloride]

To dissolve 16 g of protected polymer in 100 ml of trifluoroacetic acid, 100 ml of 4N acetic hydrobromide solution are added. By pouring the precipitate-containing solution into ether, the final product is recovered in a yield of 14.7 g (91.6%). Analysis: Br calculated: 37.7%; found: 36.8%. A _{254 nm} = 0. Paper chromatography on W1 with the developing agent n-butanol / glacial acetic acid / pyridine / water 30: 6: 24: 20 v / v).

The yield is 36.82% of the protected Monomer to the polymer and 18.85% of free lysine. The molecular weight of the polymers thus obtained is between 5500 and 20 300 daltons.

h) purification of the crude product

The aqueous solution of 250 mg according to g) obtained Polymer is loaded on a Sephadex G-50 (fine) column (80 × 4 cm) gel chromatographed. Distilled water, 0.9% by weight Saline or 0.1 N hydrochloric acid are used as Used eluent. The fractions of 1 ml are at 220 nm demonstrated.

i) Purification of the crude product

20 ml of ethanol are added to the aqueous solution of 2 g according to g) prepared ε- poly-L-lysine hydrobromide in 2 ml of water and after trituration 80 ml of ether are added and it is precipitated. The ether is changed twice by decanting. After cooling overnight, the mixture is filtered, washed with ether and dried. This gives 1.26 g (83%) of product. The yield is 57.7% calculated on the protected starting tripeptide.

In this way were also the following compounds manufactured:

SZTP-14: molecular weight 12,700 daltons calculated on the free base, [ α ] + 32.4 ° (c = 2, H₂O),
SZIP-15: molecular weight 11,600 ± 200 daltons, [ α ] + 31.9 °.
SZIP-16: molecular weight 13,400 ± 200 daltons, [ α ] + 32.6 °.
SZIP-17: molecular weight 14,500 ± 200 daltons, [ α ] + 32.1 °.

example 2 ε- poly-L-lysine hydrochloride

The solution of 1 g of polymer (prepared as in the example 1) in 40 ml of 2m sodium chloride solution is first against 2m sodium chloride solution, then dialyzed against water. The substance is obtained by lyophilization.

example 3 ε- poly-L-lysine (free base)

A column is prepared from 40 ml of Dowex 1 anion exchange resin, which is in the OH ^- cycle. The solution of 1 g of polymer (prepared as in Example 1) in 20 ml of water is allowed to flow through this column at a rate of 1 ml / minute. The fractions with a basic pH are combined and lyophilized.

example 4 ε- poly-D-lysine hydrobromide

From 10 g of (N ^α -benzyloxycarbonyl) - (N ^ε- tert-butyloxycarbonyl) -d-lysine p-nitrophenyl ester, 1.3 g of free polymer are obtained by using the method of Example 1; Molecular weight: 8900 daltons; [ α ] -47.08 ° (c = 0.92, H₂O).

example 5 δ -poly-L-ornithine

According to the method of Example 1, 10 g of N ^α -benzyloxycarbonyl- (N ^δ -tert.-butyloxycarbonyl) -L-ornithine p-nitrophenyl ester become 7.9 g of N ^α -benzyloxycarbonyl-L-ornithine p-nitrophenyl ester hydrochloride reacted, which melts at 87-90 ° C. This salt is converted with N ^α -benzyloxycarbonyl- (N ^δ -tert.-butyloxycarbonyl) -L-ornithine according to Example 1b) to N ^α -ZN ^δ - (N ' ^α -ZN'- ^δ- tert-butyloxycarbonyl-L-ornithyl ) -L-ornithine p-nitrophenyl ester dipeptide, which is obtained in a yield of 12.1 g, mp: 133-136 ° C. By repeating the procedure of Example 1a) from this dipeptide 11.99 g of N ^α -Z-N ^w - (N ' ^α -Z-ornithyl) -L-ornithine p-nitrophenyl ester hydrochloride m.p .: 115-119 ° C., and from this 14.8 g of N ^δ- Boc-tri (N ^α -ZL-ornithine) -p-nitrophenyl ester are prepared by the coupling according to Example 1b), mp: 150-153 ° C. Subsequently, according to Example 1e) tri (N ^α -ZL-ornithine) -p-nitrophenyl ester salt, mp.: 115-130 ° C, prepared, and 22 g of this salt are polycondensed in dimethylsulfoxide according to Example 1f). Thus, 15.4 g (87.6%) of δ -poly- (N ^α -benzyloxycarbonyl-L-ornithine) are obtained which, after a treatment according to Example 1g), contains 13.1 g (93%) of δ -poly-L -ornithine hydrobromide gives.

example 6 δ -poly-D-ornithine hydrobromide

The procedure described in Example 5 is used, with the difference that 1 g of N ^α -benzyloxycarbonyl (N ^δ -tert.-butyloxycarbonyl) -D-ornithine p-nitrophenyl ester is used as the starting material. Thus, 1.2 g (89%) of the title compound is obtained as the hydrobromide.

example 7 γ -poly- (L- α , γ -diaminobutyric acid)

According to the method of Example 1, 9.8 g of N ^α -benzyloxycarbonyl- (N ^ε -tert.-butyloxycarbonyl) -L- α , γ- diaminobutyric acid p-nitrophenyl ester to 6.9 g of N ^α - benzyloxycarbonyl-L- α reacted γ -diaminobutyric acid p-nitrophenyl ester hydrochloride, which melts at 67-70 ° C. This salt is converted according to Example 1b with N ^α -Benzyloxycarbonyl- (N ^γ -tert.-butyloxycarbonyl) -L- α , γ -diaminobutyric to N ^α -ZN ^γ - (N ' ^α -ZN' ^γ- tert-butyloxycarbonyl - α, γ -diaminobutyl) - L- a, γ diaminobutyric acid-p-nitrophenyl ester dipeptide coupled. This product is obtained with a yield of 10.8 g, mp: 125-127 ° C. By repeating the procedure of Example 1a) can be reached from this dipeptide to 10.35 g of N ^α - ZN ^γ - (N ^'α -ZL- α, γ -diaminobutyryl) -L-a, γ -diaminobuttersäure- p-nitrophenyl ester hydrochloride, m.p .: 108-112 ° C, which is then by a method described in example 1b) coupling 13.7 g of N-Boc ^γ-tri (N ^a -ZL- α, γ diaminobutyric acid) - p-nitrophenyl ester obtained, m.p. .: 143-145 ° C. This latter compound is reacted according to Example 1e) to tri (N ^α -Z-L- α , γ -diaminobutyric acid) -p-nitrophenyl ester salt, mp.: 105-110 ° C, the 10.5 g according to the example 1f) is polycondensed in dimethyl sulfoxide. Thus, 7.8 g of γ- poly- (N ^α -benzyloxycarbonyl-L- α , γ- diaminobutyric acid) are obtained (yield 86.5%) and from this according to example 1g) 6.2 g (90.5%) γ-poly- (L a, γ diaminobutyric acid) - hydrobromide won.

example 8 β- poly- (L- α , β- diaminobutyric acid)

According to Example 1, 7.1 g of N ^α -benzyloxycarbonyl-L- α , β- diaminopropionic acid p-nitrophenyl ester hydrochloride, mp. 65-67 ° C, from 9.6 g of N ^α -Benzyloxycarbonyl- [N ^β - tert-butyloxycarbonyl) -L- α , β- diaminopropionic acid p-nitrophenyl ester. The salt obtained is reacted with N ^α - benzyloxycarbonyl (N ^β -t-butyloxycarbonyl) -L-α, β -diamino- propionic acid according to Example 1b) to give N ^α -ZN ^β - (N ^'α -ZN' ^β - tert-butyloxycarbonyl-L-α, β -diaminopropionyl) -L-α, β -diaminopropionsäure- p-nitrophenyl ester dipeptide coupled. The yield of this coupling is 11.0 g, the product melts at 119-122 ° C. By repeating the procedure of Example 1a) can be accessed from this dipeptide to N ^α -ZN ^β - (N ^'α -ZL- α, -diaminopropionsäure- p-nitrophenyl ester hydrochloride β with a yield of 10.65 g m.p .: 102 -105 ° C. By coupling this last substance according to example 1b) 12.6 g of N ^β -Boctri (N ^α -ZL- α, β diaminopropionic acid) -p-nitrophenyl ester obtained, m.p .: 138-140 ° C , From this is prepared Tri (N ^α -ZL- α , β- diaminopropionic acid) -p-nitrophenyl ester salt according to Example 1e), mp. 102-106 ° C, yield 12.4 g. 12.4 g of this product are polycondensed in dimethylsulfoxide according to Example 1f). Thus, 8.7 g (90.2%) of β- poly- (N ^α -benzyloxycarbonyl-L- α , β- diaminopropionic acid) are obtained, which, after a treatment according to Example 1g), contains 5.6 g (88.7%). ) β- poly- (L- α , β- diamino-propionic acid) hydrobromide.

example 9 An active ingredient-containing injection solution according to the invention

500 mg of ε- poly-L-lysine hydrobromide are dissolved in 10 ml of physiological saline and the solution is sterile filtered. The thus-obtained injection solution is administered in a daily dose of 1 to 100 mg / kg depending on the condition of the patient.

example 10 A tablet composition containing an active ingredient according to the invention

Tablets are made from the following components:

10 g of polyiso-L-lysine hydrochloride
60 g lactose
16 g of sucrose
20 g of corn starch
2 g of magnesium stearate
2 g of alginic acid

The components are homogenized and the mixture is pressed to 100 tablets.  

example 11 Method for increasing the resistance of plants against virus infection

100 liters of aqueous solution are prepared from 5 g of polyiso- L-lysine produced, its pH by addition of a Sodium hydroxide solution is adjusted to 7.0 to 7.2; then 1 ml of Tween 20 wetting agent is added. The thus obtained Solution becomes after the casserole state of the plants an area of 1 hectare.

Claims (16)

1. Polyisodiaminocarbonsäuren the general formula wherein
R₄, R₅, R₆, R₇, R₈, R₉ and R₁₀ independently of one another represent hydrogen or alkyl radicals having 1 to 4 carbon atoms,
r is an average of 10 to 400,
m is a number from 0 to 10 and
n represents a number from 0 to 10,
and their salts, optical isomers and racemates. 2. Polyisodiaminocarbonsäuren according to claim 1, characterized in that
m is a number from 0 to 3 and / or
n 0 means. 3. polyisodiamine carboxylic acids according to claim 1 or 2, characterized in that the or the Alkyl radical (s), for which [n] R₄, R₅, R₆, R₇, R₈, R₉ and / or R₁₀ can stand respectively may be [a] such 1 or 2 carbon atom (s) is or is. 4. Polyisodiaminocarbonsäuren according to claim 1 to 3, characterized in that r is a number from 40 to 200. 5. polyisodiaminocarboxylic according to claim 1 to 4, characterized in that it has the L configuration to have. 6. Polyisolysine with an average molecular weight from 5,500 to 25,600 daltons. 7. Polyisoornithine with an average molecular weight from 4400 to 13,500 daltons. 8. A process for the preparation of the compounds according to claim 1 to 7, characterized in that protected monomers or oligomers with [a] diamino acid unit (s) of the general formula wherein
q is a number from 1 to 9,
R₁ is an amino protecting group and is the same in oligomers in each oligomeric unit,
-O-R₃ is an active ester protecting group which serves to protect and simultaneously activate the carboxyl group,
R₄, R₅, R₆, R₇, R₈, R₉, R₁₀, m and n have the meanings given in claim 1 to 3,
wherein in the case of oligomers, the meanings of the substituents in the individual members are the same or different, with protected monomeric diamino acid derivatives of the general formula wherein
R₂ is an amino-protecting group different from R₁ and selectively removable from R₁,
X is a group used to activate the carboxyl group, with the proviso that the group of the general formula more reactive than the group of the general formula is, means and
R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀ m and n have the meanings given above,
from the thus obtained protected oligo- or Polyisodiaminocarbonsäuren the general formula wherein
R₁, R₄, R₅, R₆, R₇, R₈, R₉, R₁₀ m and n have the meanings given above and
s is an integer from 2 to 10,
the amino protecting group R₂ splits off and from the resulting protected oligo- or polyisodiaminocarboxylic acids, in the event that s 2 to 9, after their reaction with protected monomeric diamino acid derivatives of general formula III and / or polycondensates the protecting groups R₁ cleaved off, whereupon in a conventional manner if appropriate, the resultant polyisodiaminocarboxylic acids of the general formula I are converted into acid addition salts or, if appropriate, the resulting acid addition salts of the polyisodiaminocarboxylic acids of the general formula I are converted into the free polyisodiaminocarboxylic acids or other acid addition salts and / or optionally cleavage of the resulting racemic polyisodiaminocarboxylic acids of the general formula I or salts thereof into theirs optically active antipodes or a racemization of the corresponding optically active compounds. 9. The method according to claim 8 for the preparation of Polyisodiaminocarbonsäuren the general formula I,
at which
m 3 means and
n is 0,
characterized in that as protected monomers or oligomers with [a] diamino acid unit (s) of the general formula II, those
at which
m 3 means and
n is 0,
used as protected monomeric diamino acid derivatives of general formula II, those
at which
m 3 means and
n is 0,
used. 10. The method according to claim 8 for the preparation of Polyisodiaminocarbonsäuren the general formula I,
at which
m 2 means and
n is 0,
characterized in that as protected monomers or oligomers with [a] diamino acid unit (s) of the general formula II, those
at which
m 2 means and
n is 0,
used as protected monomeric diamino acid derivatives of general formula III, those
at which
m 2 means and
n is 0,
used. 11. The method of claim 8 or 9 for the preparation of polyiso-L-lysines, characterized in that as a protected monomer with [a] Diaminosäureeinheit (s) of the general formula II N ^α -ZL-lysine p-nitrophenyl ester hydrochloride and as a protected monomeric diamino acid derivative of general formula III, a mixed anhydride obtained from N ^α -Z-N ^ε -Boc-L-lysine with isobutyl chloroformate. 12. The method according to claim 8 or 10 for the production of polyiso-L-ornithines, characterized in that as a protected monomer with [a] Diaminosäureeinheit (s) of the general formula II N ^α -ZL-ornithine p-nitrophenyl ester hydrochloride and as a protected monomeric diamino acid derivative of general formula III, a mixed anhydride obtained from N ^α -ZN ^β -Boc-L-ornithine with isobutyl chloroformate. 13. The method according to claim 8 or 9 for the preparation of polyiso-D-lysines, characterized in that as a protected monomer with [a] diamino acid unit (s) of the general formula II N ^α -ZD-lysine p-nitrophenyl ester hydrochloride and as a protected monomeric diamino acid derivative of general formula III, a mixed anhydride obtained from N ^α -ZN ^ε -Boc-D-lysine with isobutyl chloroformate was used. 14. The method according to claim 8 or 10 for the preparation of polyiso-D-ornithines, characterized in that as a protected monomer with [a] diamino acid unit (s) of the general formula II N ^α -ZD-ornithin- p-nitrophenyl ester hydrochloride and as the protected monomeric diamino acid derivative of the general formula III, a mixed anhydride obtained from N ^α -ZN- ^β -Boc-D-ornithine with isobutyl chloroformate was used. 15. Medicinal products characterized by a content 1 or more compound (s) according to claims 1 to 7 as active ingredient (s), optionally together with 1  or more conventional pharmaceutical carrier and / or Excipient (s). 16. Plant protection products, in particular for increasing the resistance of plants to virus infections, characterized by a content of 1 or more Compound (s) according to claims 1 to 7, optionally together with 1 or more in plant protection products customary carrier and / or excipient (s).