JPH02124861A - Isopolypeptide and production thereof - Google Patents

Abstract

NEW MATERIAL: A compound of formula I (wherein R⁴ to R⁷ and R¹⁰ are each H or a 1-4C alkyl; R⁸ and R⁹ are each H; r is 10-400; m is 0-3; and n is 0-10), a salt thereof, a recemic modification thereof and an optical isomer thereof.

EXAMPLE: Isopoly-L-lysine having an average molecular weight of 5,500-25,600.

USE: Medicines. An active ingredient of antitumoral and antiviral agents. Useful in the treatment of a cancer.

PREPARATION: A monomer or oligomer of formula II (wherein q is 1-9; R¹ is an amino protective group; R³ is an active ester protective group for the protective group and the carboxyl group) is coupled with a monomer of formula III (wherein R² is am amino protective group different from R¹ and capable of being selectively removed independently of R¹ (the -COX is more reactive than the COOR²). Protective group R₂ is removed from the obtained compound, and the deprotected compound is reacted with a monomer of formula III or polycondensed to obtain a compound of formula I.

COPYRIGHT: (C)1990,JPO

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to novel isopolypeptides of diaminomonocarboxylic acids, salts, optical isomers and racemates thereof, and methods for producing these compounds.

Particularly, the present invention relates to a novel diaminomonocarboxylic acid isopolypeptide having the following formula (1).

(In the formula R4, ns, R6, R7, R1, R' and R1
l1t: [each independently a hydrogen atom or an alkyl group of C1~ (hereinafter referred to as a lower alkyl group): r is the average value lG
400 to 400; m is an integer of O to 10; n is an integer of O to 10) Note that the contents of the corresponding substituents in each term do not necessarily have to be the same and may be different.

BACKGROUND OF THE INVENTION Isopeptides of diamino-monocarboxylic acids are rare in nature and are found in certain peptide antibiotics, in bacterial cell walls, in cross-linking of some proteins, and in clavicepamine. It's just a matter of getting caught. Some of these exhibit biological effects and can be produced by microbiological methods.

Japanese researchers have developed an L-lysine polymer consisting of 25 to 30 monomer units linked together by isopeptide bonds to Streptomyces alplus (streptomyces alplus).
ptomyces albulus) [Sima and Sakai: Agricultural and Biological Chemistry (Agr.
ic, Biol, Chew,) 45.2503(
(1981)), and this poly-ε-L-lysine inactivates various bacterial phages [Sima et al.: Agric;
Biol, Chew, 46°1917 (198
2)), whose biosynthetic approach has been applied for its production (Shima et al.: Journal of the Japanese Society of Agricultural Chemistry 57, 221-2
26 (1983)].

Clavicepamine is a synenriched basic protein isolated from saprophytic cultures of the ergot fungus and has pharmacological importance for humans. These proteins are 30
~95 mol% high lysine content, molecular weight, 2000-17
,000 compounds. Biological studies on these compounds have shown that these compounds have cell growth-retarding effects and suppress tumor growth in animals without substantial toxic side effects [TIHAC (
TyihAk): Dissertation, 1978
).

It was established that the C-lysine (poly)peptide is the basic structural unit of clavicebamine based on its structural determination [Sujikan (6n to Sz6), Kelemen (Kelemen),
men) and Terek (T8r8k): Journal of Chromatography (J, Chromatogr, ),
366, p. 283 (1986)). Until now, the role of the polyisolysine structure in the promising biological effects of clevicepamine was unknown.

The antiviral activity of synthetic poly-L-isolysine, its acylated derivatives with amino acids, and poly-β-L-α, β-diaminopropionic acid was determined by researchers in India. As a result, the antiviral activity of these polypeptides was found to be related to their ability to induce interferon-like antiviral factors during CAM culture [Indian Journal of Experimental Biology] (Indian J, Exper, Bi
20. No. 227 (1982); Indian Journal of Chemistry (Indi
an J, Chew, ) 21B, p. 483 (19
82)).

DISCLOSURE OF THE INVENTION In order to study the relationship between chemical structure and biological action, the present invention uses the It (MW) and polydispersity (MW) and polydispersity (10 to
The aim was to produce on an industrial scale an isopeptide of a monodiaminomonocarboxylic acid having a molecular weight of 400 units).

The basic requirements for this given compound were high purity, low polydispersity and high stereochemical purity.

It is known that the action of biopolymers is greatly influenced by their molecular weight distribution. The arrangement of amino acids is also very important. This is because different optical isomers may have opposite and completely different biological effects. For example, D-arginine suppresses Illll in animals, but L-arginine stimulates it [
Szende and TyibAk
): Lancet 7546, No. 8
24 pages (+968); Szende: D
issertation, 1974). The racemic mixture or racemized polymer no longer exhibits its original favorable biological effects.

It is desired that the above polypeptide has high purity and low polydispersity, and a synthetic method was chosen to solve this problem. Hull's method [pyopolymer (B)
iopolymers) 17.2427 (1978)
α-Z-protected polysine active esters were further prepared by the method of Gangopadhyay and Mathur [Indian Journal
of Chemistry (Indian J, Chem,
) 21B, p. 483 (1982))
-BOC-protected synpentachlorophenyl-esters were each polycondensed. Nishi's method [International Journal of Biological Macromolecules (Int. J. Biol.
, Macromol, ) 2. Page 53 (1980) α-Z-Lys-OH was polymerized using diphenylphosphoryl azide (DI'PA). This document contains information on molecular weight (MW), monodistribution and its gloss purity. No data are available. However, in the experience of the present inventor, only those with low I (up to 2500) and high polydispersity were obtained in low yields, and these had no biological effect. Kushatia and co-workers' synthesis method [Biopolymer
), p. 219 (1980)) and Mercer (Mat.
hur), etc. (International Journal of Peptide Protein Research (Int,
J, PeptideProt, Res, ) 17.
189 (1981)) appears to be the most suitable for obtaining products with low dispersion.

ε-poly-lysine and δ-poly-ornithine have been synthesized by them. The principle of this method is the same for both compounds. That is, starting from a protected active ester or an azide-protected amino acid methyl ester, a tripeptide-methyl ester is first obtained. After alkaline hydrolysis, this tripeptide acid is then activated into an active ester form (which is converted to pentachlorophenyl ester). After deprotection of the ω-7 amino terminus, this tripeptide is easily polycondensed. Finally, the α-amino protecting group is removed and the isopolypeptide is separated. In these methods, BOD at the α-position,
A 2-amino protecting group is used in the α-position. Removal of these protecting groups is carried out by treatment with a strong acid and catalytic hydrogenation, respectively. In the above-mentioned literature, the polymers are not characterized by molecular weight distribution or optical purity.

The abbreviations used herein are based on the recommendations of the IUPAC-IUB Committee on Biochemical Nomenclature [Journal of Biology and Chemistry].
J, Biol, Chell, ) 247. No. 977
(1972)), with further symbols added and summarized below.

2: Benzyloxycarbonyl BOC: tert-butyloxycarbonyl-0
Np: P-nitrophenyl-05n: N-hydroxy-succinimide-0
Pcp: Pentachlorophenyl-0Pfp: Pentafluorophenyl-0Bt: N-hydroxybenzotriazolyl BuOCO: Isobutyloxycarbonyl EtOCO: Ethyloxycarbonyl N
,: Azide EEDQ: N-ethoxycarbonyl-2-ethoxy-1°2-dihydroisoquinoline 1! tOAc: Ethyl acetate IIDQ: N-isobutoxycarbonyl-2-isobutoxy-1,2-dihydroisoquinoline THF:
Tetrahydrofuran DMF: Dimethylformamide) IPLc: High performance liquid chromatography 0PTL
C: Overpressure thin layer chromatography IEF: Isoelectric flux MS: Mass spectrometry NMR: Nuclear magnetic resonance spectroscopy Zco,: 2- measured from released CO2
Group Mv = Molecular weight TLC: Thin layer chromatography BuOH: Butanol AcOH: Acetic acid When reproducing the above method, several drawbacks were found.

That is, in the production of oligomers, the carboxyl groups are always protected with methyl esters, and therefore the oligomers obtained are not suitable for polycondensation as they are. This step, which must be liberated from the ester, results in racemization as well as other side reactions, which reduce the yield. Furthermore, unfortunately the polydispersity of the synthesized polymers is high and the application of the benzyloxycarbonyl-ω-amino protecting group is disadvantageous since it is removed by catalytic hydrogenation. Furthermore, unfortunately, the polymers thus obtained were found to be ineffective in the biological tests contemplated by the present invention.

The present invention therefore aims to provide a new method which can eliminate the drawbacks of the conventional methods as mentioned above, and which has not been previously attempted for the production of isopeptides of diaminodicarboxylic acids. There wasn't.

The present invention is based on the discovery that the above object can be achieved by the method described below. That is, first an active oligomer is prepared that can undergo direct polycondensation, 6 in which the carboxyl group is protected with an active ester group, and peptide bonding occurs at a rate significantly faster than the aminolysis of this active protected ester group. The inventors have now found that a backing off activation method is well suited for producing isopeptides and their salts containing diaminocarboxylic acids. C0OH
For temporary protection and simultaneous activation of groups, ρ-nitrophenyl esters are preferred, and for peptide bonds the mixed anhydride method is preferred, whereby the amino group in the subsequent amide bond is protected with a BOC group, Amino groups that do not participate in the reaction are protected with two groups. In this way.

Homo- or sequential isopeptides of any diamino-carboxylic acid can be synthesized. Furthermore, the inventors have discovered that isopolypeptides of various molecular weights can be produced reproducibly with low polydispersity. The resulting intermediates and final products have good crystallinity, high purity, and can be easily isolated.

The present invention is further based on the following findings.

Thus, in the case of diaminocarboxylic acids, p-nitrophenyl ester is the most convenient group for one-time protection and simultaneous activation of the carboxyl group, the mixed anhydride method is best suited for peptide bonding, and the Z group and The BOC group is selectively used for protection of other groups.

Finally, the present invention is based on the following findings. That is, any homo- or sequential isopeptide of diamino-monocarboxylic acid can be synthesized by the above method, and its morphology is maintained without racemization.

The present invention provides isopolypeptides of the above general formula (■) (substituents are as defined above), salts, optical isomers and racemates thereof.

According to the method of the present invention, the compound of general formula (1) and its salt are produced as follows.

Monomers or oligomers of the following general formula (II): (wherein q is an integer from 1 to 9; R1 is an amino protecting group (all R1s are the same in the case of oligomers); R1 is suitable for protection as well as activation Tameno active ester protecting group for carboxyl group; R4, RG, RG, R7, R8, R'', R''
, m and n are the same as above; however, in the case of oligomers, the substituents may be the same or different.) Monomer of the following general formula (III): (wherein, R2 is an amino protecting group and R1 is different and 8
X is a carboxyl-activated group (however, the -COX group is more reactive than the COOR3 group);
', R'', R', R'', m and n are the same as above) to obtain a compound of the following general formula (IV): (wherein, all substituents are the same as above; S is an integer from 2 to 10) and then, if S≦9, the resulting compound is reacted with a monomer of general formula (III) or polycondensed by conventional means. The R1 protecting group is then removed from the product, optionally converted into a pharmaceutically usable salt if the compound of general formula (1) obtained is a free base, or if it is a salt it is converted into a free base or another It can be converted into salt.

Optical isomers can be obtained by using the corresponding isomers as starting materials. If it is desired to obtain a racemic polymer, the racemic form of the monomer should be used as the starting material. In the method of the invention, R1 and R2 are selectively removable protecting groups well known in peptide chemistry.

A preferred R1 is a benzyloxycarbonyl group (Z), and a preferred R2 is a tertiary-butyloxycarbonyl group (BOC). -OR' group is an active ester group for both protection and activation of carboxyl groups, such as -0Np, -0Sn, -0Pcp, -0P
fp, -0at.

X is BuiOCO-1EtOCO-, N, -0Sn,
-0Pcp, -0Pfp, and specifically depend on the reaction conditions and the content of R3, such as BuiOCo- or Et
It is an OCO- group. The monomers of general formula (II) used as starting materials in the most advantageous process of the invention are those in which R1 is Z and R'' is BOC1R3 10Np.7 This BOC group is removed by acidolysis and then protected. diaminocarboxylic acid (R”=Z
= R''=BOC) is coupled by the mixed anhydride method.

After removing the BOC protecting group, the resulting dipeptide is polycondensed or coupled again with the next protected amino acid as described above, and the process is repeated to give the decapeptide.

The best method is to repeat the above linkage to form a tripeptide and convert the resulting oligomer (suitable for direct polycondensation) into an isopolypeptide with a predetermined number of amino acid residues. Removal of the 2-protecting group (ie, deprotection) is preferably performed with HBr in acetic acid and the final product is isolated.

The most advantageous processing conditions for the mixed anhydrous bonding method are as follows.

(a) Solvents niacetonitrile, THF, DMF, dichloromethane; (b) Reagent-forming anhydrides: ethyl chloroformate, isobutyl-chloroformate, EEDQ, IIDQ; tertiary bases nitriethylamine, diisopropyl-ethylamine, N -Methyl-morpholine: (c) Activation time and temperature: 10-30 minutes, -10°C to -20°C; (d) Neutralization of the amino component salt: using triethylamine or diisopropyl-ethylamine in the reaction mixture.

Yield of pure final product: 60-80%.

Both homo- and sequential-isopolypeptides can be made with the methods of the invention.

For analysis of the produced isopeptides, the latest separation techniques such as HPLClOPTLC and IEF were used, as well as IR, MS, and NMR as determination methods. These studies and controls verified the chemical structure.

For the investigation of biological activity, the purified substances were used as described in the examples. That is, the purification was carried out by precipitation, column and gel chromatography.

As a result of biological tests, poly-L-isolysine produced by the method of the present invention [degree of polymerization (p, d,): 10-400]
9 of them have a degree of polymerization of 40 to 200 (molecular weight: 5500 to
The present inventors have found that the compound (20,300) has a cell proliferation inhibitory effect, that is, an antitumor activity in vivo and in vitro, and can suppress tumor metastasis6.
200 to 15,400 (degree of polymerization = 80 to 120) had the greatest effect. Other available substances have so far not been found to have any effect in this respect. The reason is primarily due to the high polydispersity and racemization provided by the Kushava or Mathur methods.

Poly-L-isolysine with a degree of polymerization of 80-120 prepared by the Kushawa or Hull method did not exhibit any antiviral activity.

The main advantages of the synthesis method of the present invention compared to conventional methods are as follows.

Products with promising biological effects can be manufactured with good reproducibility.

It is possible to obtain oligomers which can be directly polycondensed, which makes it possible to dispense with the aforementioned hydrolysis step and also to avoid racemization, which is very dangerous from the point of view of biological effects.

In a wide range of molecular weights, products with favorable molecular weight distributions (i.e. low polydispersity) are obtained, which is also advantageous for achieving the desired biological effect (e.g. in the case of polymer 5ZTP-14). , molecular weight is 12,700±2
00; polydispersity: 3.7). Yields are also high, and the purity of intermediates and final products is also high. Furthermore, it is easy to crystallize and isolate. There are no particular difficulties in the reproducibility of the process.

The reaction time in the peptide coupling step is also short.

The combination of amino protecting groups of the present invention makes it possible to omit deprotection by catalytic hydrogenation. This is extremely advantageous in production on an industrial scale.

The most important advantage of the process of the invention is the application of a racemization-free reaction step.

As mentioned above, the products obtained by the conventional method were not found to have any effect in the biological tests conducted by the present inventors. In contrast, the polymer synthesized according to the present invention showed very promising biological activity.

The following matters are new in the compound of general formula (1) obtained by the method of the present invention.

R4, R%, RG, R? and R1@ are each independently a hydrogen atom, a methyl group, an ethyl group, a propyl group, or a butyl group; R'' and R' are each a hydrogen atom; r is an integer of 10 to 400; m is 6. 1.2 or 3; n is O.

In the case of a preferred group of compounds: R4, R5, RG, R' and R'' are each a dihydrogen atom, a methyl group, an ethyl group, a propyl group or a butyl group; R1 and R'' are each a hydrogen atom r is an integer from 40 to 200; m is 3; n is 0.

In the case of other preferred compound groups: R4, R8, RG, R' and R'' are hydrogen atoms; R'' and R' are hydrogen atoms; r is an integer from 80 to 120; ; m is 3; n is 0.

In yet another preferred group of compounds, two R4, R'',
R', R7 and R111 are each independently a hydrogen atom, methyl group, ethyl group, propyl group or butyl group; R8 and R9 are each a hydrogen atom; r is an integer from 1O to 120; m is 2 So; n is 0.

In yet another preferred group of compounds: R4, R5, R@, R7 and R10 are each independently a hydrogen atom, methyl group, ethyl group, propyl group or butyl group; R1 and R9 are each a hydrogen atom; is an integer from 10 to 400; m is 1; and n is O.

In yet another preferred group of compounds: R4, R5 and R10 are each independently a hydrogen atom, methyl group, ethyl group, propyl group or butyl group; R5, R6, R1 and R9 are each a hydrogen atom:
r is an integer from 10 to 400; m and n are O.

It has been found that the L-stereoisomer of the isopolypeptide of general formula (1) and its salts are highly effective against tumors and viral diseases.

The biological activity of polyisolysine produced by the method of the present invention was tested in vitro and in vivo.

Promising effects have been demonstrated in the following respects. Note that the antitumor effect was measured on animal tumor lines, and the antiviral effect was tested on plant viruses.

In these experiments, the molecular weight ranged from 5500 to 25,60
It has been found that isopoly-L-lysine with a molecular weight of 0 is extremely effective, especially those with a molecular weight of 10,200 to 15,400.

In both measurements, substance 5ZTP-14 was the most effective. Its characteristics were as follows.

-z-Viscosity of protected polymer: Positive = 31.92 cd/g
,・Zeo, : 16.8% (calculated value); 16.3
(Experimental value).

Free polymer was obtained as HBr salt.

・Br component: 37.7% (calculated value), 36.8% (
experimental value).

- A2□; 0 (ie, UV absorption is zero).

Purification was carried out by precipitation from alcohol using ether.

・[α]ra'=+32.4'(c=2; water) ・Molecular weight=12,700±200 Cultivation is used, and in the case of 1st theft, animal species that can be translocated are used. Accordingly, the compounds of the present invention can be used in human erythroleukemia cell lines (K-562, Karolinska Inst.
5titut), Stockholm], and four transplantable murine tumors (I12x. Lymphocytic leukemia, PjmD macrophage-induced tumor induction - Ehrlich)
Cancer, Lewis Lung Tumor] @ I-txt. In the case of Ehrlich carcinoma and R3° tumor, the number of surviving animals was used as an indicator of efficacy. LL
For T, the number and size of metastases (in the lungs or liver sac)
was determined for treated and untreated mice. This experiment consisted of an injection solution, i.e., each compound was mixed with 10 mL of saline.
The test was carried out using a solution of 50 mg dissolved in G.

The compounds tested were labeled as follows.

5ZTP: Polyiso-L- produced by the method of the present invention
Lysine; 5ZTP-14: polyiso-lysine HBr salt, molecular weight calculated based on free base: 12,700±2
00; Polydispersity: 3.7 (number average/weight average); 5Z
TP-15: Polyiso-thi-lysine HBr salt, calculated molecular weight based on free base: 11,600±200
;5ZTP-16: Polyiso L-lysine HBr salt, molecular weight calculated based on free base: 13,400±
200; 5ZTP-17: Polyiso L-lysine HBr
Calculated molecular weight based on salt, free base: 14,5
00±200; 5ZTO: polyiso-1,-ornithine produced by the method of the present invention; 5ZTO-18: polyiso-1,-ornithine) lBr
Calculated molecular weight based on salt, free base: 8, 900±200 Nid 17: Polyisolysine manufactured by Hull et al.; Ni-17: Polyisolysine manufactured by Kushawa et al.; A-3: α-polylysine.

Summary of Experimental Results Regarding Tumor Cell Increase The compound produced by the method of the present invention reduced the number of tumor cells in vitro, although it was dose dependent, compared to the control. This effect appeared as a stagnation of cell numbers when the dose was low, but when the dose was high it killed the cells. H-17 and To-17 had no effect at all. α−
Polylysine (A-3) only showed stagnation in cell number.

In in vivo experiments, treatment with 5ZTP resulted in increased survival of treated animals.

In the case of Ehrlichoma, this treatment resulted in a very long tumor-free period, with tumor growth seen only after treatment was stopped.

As a result of treatment with 5ZTP, metastasis to the liver can be completely prevented even when the tumor is inoculated into the elephant.
A reduction in the number and size of lung metastases was also observed when the tumor was inoculated intramuscularly.

Therefore, it has been demonstrated that polyisolysine and polyisoornithine produced according to the present invention can inhibit tumor cell proliferation. Among these compounds, polyisolysine in particular showed an extremely strong inhibitory effect. This inhibitory effect was comparable with the best conventional cytostatic drugs in terms of animal survival. The compounds of the invention also clearly demonstrated excellent anti-metastatic activity. Another advantage of the present invention is its low toxicity, which is significantly lower than that of conventional cytostatic drugs.

For example, according to the biological tests of the present invention, the therapeutic index: (maximum tolerated dose/minimum effective dose) is greater than 10, whereas most conventional cytostatic drugs are usually much greater than 10. It is below.

In addition, there is no difference in biological response between males and females,
This is also an advantage.

Data for explaining the antitumor effect will be described below.

Tables 1a to 1C show polyisolysine produced by various methods,
Polyisoornithine produced according to the present invention.

The effect of α-polylysine on the proliferation of Ni-562 cells is shown.

The experimental parameters are as follows.

Cells: K-562 human erythroleukemia cell line [Karolinska I
Institute), Stockholm] was used.

Cell count: 5 X to'/mQ (a glass test tube containing 5 to 6 IIIQ medium was used).

Number of tests = 3 test tubes were used per administration.

Medium = Parker's (P) supplemented with 10% fetal bovine serum
M-199 [Flow Laboratories, Scotland].

Temperature and atmosphere: 37°C, 5% CO□ + 95% air.

Treatment: 24 hours after dilution of cell culture medium.

Dose: 1-10-100 μg/mQ.

Evaluation: Cell counting was performed using a Buerker's chamber 24 hours, 48 hours, 72 hours, and 96 hours after dilution of the medium.

Number of experiments: 2 per compound.

Table 1a shows 5ZTP-,14,5ZTP-15,5ZTP
-16,5ZTP-17 and 5ZTP-18 are shown to have a large dose-dependent inhibitory effect on cell proliferation. What is noteworthy here is that treatment with a dose of just 1 μg/IIIQ resulted in a clear antiproliferative effect.

Table 1b shows, on the contrary, that it showed no effect on K-562 cell proliferation.

Table 10 shows that α-polylysine exerts a plateau in cell numbers when compared to the control.

However, even high doses (100 μg/ml) did not result in an increase in cell number.

The values in these tables are at the given times.

Cell numbers were counted in three tubes for each treatment and control.

Table 1a (Effect of compounds of the invention on Ni-562 cell line in vitro) Treatment: 1-10-100 μg/+aQ, 2 after culture dilution
4 hours.

Table 1a Number of cells x 1037mQ (nutrient solution) SZTP-1448 hours 72 hours 96 hours 100 μg/mQ 0 0
010μg/m12 90
90 1501μg/laQ Control 5ZTP-15 100μg/rnQ 107A g/1aQ 1 p g/mQ Control SZTP-16 100μg/lllQ 10μg/mQ 1μg/mQ Control 5ZTP-17 100μg/mQ 10μg/m12 1μg/mQ Control 5ZTP-18 100Pg/+mQ 10μg/mQ 1μg/mQ Control table 1b Fl-17 against 562 cell proliferation
Effects of treatment with and -17.

Treatment: 1-10-1007 A g/mQ, 24 hours after medium dilution.

L" Cell number x 10'/mQ (nutrient solution) 48 hours 72 hours 96 hours -17 100μg/mQ 125 23
010 pg/mQ 120
2201μg/mQ 130 2
-17 to 40 100ug/mQ 120 22
01101t/d 125 24
01μg/mQ 140 240
Control 120 210 Table 1
0 Effect of A-3 treatment on Ni-562 cell proliferation in vitro.

Treatment: 1-10-100 pg/mQ, 24 hours after medium dilution.

Table 10 Number of cells x 1037mQ (nutrient solution) 48 hours 72 hours 96 hours 100μg/r
nQ 110 110 100
10μg/mQ 130 120
1201μg/mQ 110
110 100 control 120
250 300 In-Vivo Test (a) Test for LL2111 tumor, 5ZTP-14 Tumor source: Chester Beatty Institute (Ch
Ester Beatty In5titut)+London.

Animal species: DBA72 allogeneic breeding, self-breeding.

Animal weight, sex = 20-23 g, female, 5 per group Number of tumor cells: 10' per animal, intraperitoneal inoculation.

Treatment: 24 hours after tumor inoculation, the drug is administered intraperitoneally at a dose of 50 μ/kg once a day for 8 days.

Control group: 0.2 mQ of physiological saline was administered intraperitoneally every day.

Evaluation: Animal Survival Results: As shown in Table 2, the survival rate of treated animals was better than that of the control, for example, only one control mouse survived 10 days after tumor inoculation. All treated animals survived;
The first death of treated animals occurred 13 days after tumor inoculation, and the last death occurred 15 days after tumor inoculation.

Table 2 (I, □. Effect of administration of drug at 50 kg/day on the survival of DBA/2 mice inoculated with JI tumors)
BDF/1 hybrid, self-breeding.

Animal weight, sex = 20-23 g, male, 5 animals per group.

Number of tumor cells = 2,8 x 10' per animal, inoculated intraperitoneally.

Treatment: 24 hours after tumor inoculation, 10μ/k daily
g was administered intraperitoneally.

Control group: 0.2 mQ of physiological saline was administered intraperitoneally every day.

Evaluation: Number of surviving animals Results: Table 3 shows the number of surviving animals. All treated mice survived beyond control mice.

(c) Ehrlich ascites tumor, treated with 5ZTP-14 Animal species: Swiss, non-allogeneic, self-breeding.

Animal weight and sex: 20 g, 5 males, 5 females Number of tumor cells: 106 per animal Treatment W: 50 g each day starting 24 hours after inoculation of j1 tumor
■/kg and 75 ■/kg were administered intraperitoneally for 20 days.

Subject: 0.2 ml of physiological saline was administered intraperitoneally every day.

Assessment: Daily weight measurements during life.

Animals that survive 55 days after tumor inoculation are sacrificed and dissected, ascitic fluid volume is measured, and occasional solid tumor formation within the peritoneal cavity is noted.

Results: Table 4 shows the body weights of mice 16 days after tumor inoculation. Control mice died from day 17 to day 22 with a one weight excess, indicating the formation of neoplastic ascites. All but one group of treated animals survived 55 days after tumor inoculation.

These animals were then sacrificed and examined for the amount of ascites and the presence of solid tumors within the tumor. This is shown in Table 5. As a result, no tumor was found at all, but 520
Of the 2 treated mice, 16 survived 33 days longer than the last of the control mice.

Table 4 [Effects of intraperitoneal administration of 5ZTP-14 at 50 μ/kg and 75 μ/kg (20 days) on body weight and survival (when male and female Swiss mice were inoculated with Ehrlich ascites tumor; weight is 20g)
), in the table, "O" means death due to tumor; [+] means death due to other reasons.

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2024272425.5 (Continued from Table 4) 30.526313031 26283243 + Human-Dan (Swiss mice inoculated with Ehrlich ascites tumor, administered 5ZTP-14 at 50 μ/- and 75 mg/kg, and killed 55 days after tumor inoculation) Treatment (a) Lewis lung tumor (ILT), inoculated into the lung.

Animal species: Homologous McstBx, LATT, Godlo (G
5d811δ), body weight and sex of Hungarian animals: 20-23 g, female (6 controls;
(4 treated animals).

Number of tumor cells: LTT cells inoculated into lung: 5xlO'
.

Treatment W: Starting 24 hours after inoculation, administer 75 μl daily.
Administered for 8 days.

Evaluation: Animals are sacrificed 9 days after tumor inoculation and counted for liver metastases.

Results 8 Table 6 shows the number of metastases. Not a single liver metastasis was found after treatment with 5ZTP-14.

Human - (Effect of treatment with 5ZTP-14 on liver metastases in female C, Bl mice inoculated with ILT into the lungs (intraperitoneal administration = 75, /kg for 8 days)] Number of liver metastases Control 31 20 63 36 3
8 56 (ma=40.66; s:16.0
2; n=6) SZTP(40000(II=4
) Animal species: C, 7B1 allogeneic LATT, Godlo (G8d
811δ), Hungarian animal weight and sex: 20-22 g, female.

Number of animals: 5 per group.

Tumor cells: 5 x 10' cells per animal, intramuscularly administered to the rightmost extremity of the right thigh muscle. Ten days after tumor inoculation, the tumorous edges were removed. After this treatment, 111 days after vaccination 1
During the 77th day, 5ZTP-14 was administered intraperitoneally at a rate of 50@/kg. Physiological saline was intraperitoneally administered to the test animals at a dose of 0.2 μa daily.

Evaluation: 188 days after tumor inoculation, animals were sacrificed and the number and average size of lung metastases were determined using a stereomicroscope.

Results 8 Table 7 shows the number and average size of lung metastases. Treatment with 5ZTP-14 resulted in a significant reduction in the number and average size of lung metastases.

-I [5ZTP-1 on the number and average size of lung metastases in C, Bl mice administered intraperitoneally with Lewis lung tumors
Effect of treatment with 4 cg (50 μ/day daily between days 111 and 177 after tumor inoculation) was administered intraperitoneally. ]Contrast
63.13±27.53 49.45±29.07S
ZTP-1426,291,18,8720,lO+3
5.38 Intact tobacco plant [Nicotiana tabacum C.
V. Samsan (Nicotiana tabacum)
CV, Samsun)) and tobacco mosaic virus (
TMV) susceptible to infection were mechanically inoculated. In this case, carporundum powder (particle size: 500 mesh) was used as the abrasive. The inoculum contained 200 rg of resistant tobacco mosaic virus (TMV 01 strain) in 0.1 M Soelensen phosphate buffer (pH 7.2). 3 hours after inoculation, as a collection spray.

Intact leaves were treated with 5ZTP-14. This treatment was repeated for 3 consecutive days. The severity of symptoms was investigated 2 weeks after vaccination. As leaf samples, leaves of all ranks of the inoculated plants, ie, leaves of different heights and leaves of various ages, were collected 14 days after inoculation.

Virus concentration was determined from 1 g of frozen samples.

The leaf sample was uniformly dispersed in 4 mQ of phosphate buffer (p)l 7,2
) Centrifugation (approximately 10,000 r, ρ, m for 5 minutes) was performed. 15 μQ of this supernatant was used for quantitative analysis. Using rocket immunoelectrophoresis, further specific anti-TM
The virus content was estimated using W (old) serum (1 μQ of immunoglobulin/13). Sodium barbiturate buffer (0.2A, pFl 8.6) was used as the running buffer.

The virus content was calculated from the height of the precipitated peak using a standard graduation curve.

Sac-”- (Approximate example of antiviral effect) Treatment with SZTP-14 Virus content 200
mg/fi 11+ng/mQ20■I
Q 24.6■l-2mg/12
29.3■lmQ control 5
1.3 lmQ The isopolypeptides according to the invention can be used as active ingredients in pharmaceutical compositions both in the form of free base and non-toxic salts. In this case, the isopolypeptide is used in admixture with carriers and auxiliaries commonly used in the pharmaceutical industry. Furthermore, the compounds of the invention can be used in both free base and salt form to increase the resistance of plants to viral infections. In this case as well, it is used in combination with adjuvants commonly used for plant protection.

Pharmaceutically acceptable salts include acid addition salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, oxalate, fumarate, gluconate,
tartrate, malate, acetate.

Citrates, benzoates, alcoholates, lactates, alginates can be used.

Pharmaceutical products containing the compounds according to the invention as active ingredients can be used in the form of injectable solutions, tablets, sugar-coated tablets, herbal medicines, suspensions, drinking solutions, powders and the like.

The pharmacologically effective dose of the compound of the present invention depends on the method of application, patient condition, etc., but is generally administered at a rate of 0.02 to 20 cm/kg body weight/day divided into 2 to 4 times a day. It will be done. In the case of oral administration, the dosage will be higher.

An advantageous drug form is an injectable solution, which can be administered intravenously, subcutaneously, intramuscularly, etc.

In the preparation of this injection solution, isotonic NaCl solution can be used in combination with phosphate buffer, ascorbic acid stabilizer, etc., if desired.

The active substance can be administered as a pharmaceutical composition in solid or liquid form. In the case of a liquid, sweeteners, aromatics, etc. may be added.

Solid oral dosage units are preferably used in the form of tablets, dragees, capsules and the like. When manufacturing these drugs, the above active ingredients are mixed with a common pharmaceutical carrier, and the mixture is pressed into tablets, sugar-coated tablets, or capsules are usually used. Fillers, additives such as milk sugar, magnesium stearate, sucrose.

Talc, stearic acid, gelatin, candenum, pectin, gum tragacanth, colloidal silica gel.

Corn starch, alginic acid, etc. can be used.

For the treatment of plants, the compounds of the invention are combined with the usual adjuvants,
Conversion into solid or liquid formulations can be carried out, for example using wetting agents, solid carriers such as kaolin, and preferably in combination with further active ingredients. In this case, the preparation prepared in this way is applied as a solution or suspension containing the compound of the present invention at a pH of 1 to 50 pph to plants by a known method, preferably at a rate of 0.5 to 5 g/ha. do.

(Examples) The present invention will be described below based on Examples, but the present invention is not limited to these Examples.

(a) N''-benzyloxycarbonyl-shi-lysine-p-nitrophenyl ester/HCI: 10 g of No-benzyloxycarbonyl-(NE-tert-butyloxycarbonyl)-L-lysine-p-nitrophenyl ester was It was dissolved in 40 mQ of fluoroacetic acid and to this was added an equal volume of 2N HCl dissolved in EtOAC. After standing for 1 hour at room temperature, the solution was evaporated to half the volume in vacuo. Ether was then added to this to crystallize the solution.

Charge: 8.3 g (96,06%); melting point (m, p
): 83-86°C; TLCRt=0.68 (II
-butanol-acetic acid-water = 4:1:1).

(b) N''-benzyloxycarbonyl-N-(N'
"-benzyloxycarbonyl-N'ε-tert-butyloxycarbonyl-shi-lysyl)-L-lysine-P-nitrophenyl ester: No-benzyloxycarbonyl-(NE-tert-butyloxycarbonyl)-L-lysine 7.69 g (20
Milmol) was dissolved in anhydrous acetonitrile. Then, 2.8 mQ (20 mmol) of triethylamine,
Furthermore, 2.8 tQ of isobutyl chloroformate was added dropwise under vigorous stirring.

Then, after activation (20 minutes), 9 g of the salt obtained in step (a) above were added, and to this was added dropwise a cold solution of 2.8 Q of triethylamine in 5 mQ of acetonitrile. Next, stirring was continued for 2 hours at -10°C. This mixture was diluted with 0.5M HCI 300@Q. The resulting solid product was collected, dried and recrystallized from a 1:1 mixture of ethyl acetate and petroleum ether. Yield: 12
．． 9g (89,02%); ■, p: 128-13
1°C 0 analysis value: -0NP content: 99.8%. T.L.
CR,=0.64 (EtOAC-cyclohexane=3
: 1), Rf=0.91 (II-butanol-acetic acid-water=4:l:1); HPLCtr, =8.
8 minutes (K'=2.2); Column: 0DS-Hypersil-6 (125X4mm); Eluent: MaOH-CH3CN-0,02M-NaOA
c buffer (pH 4) = 40: 30: 30 (v/
v); Flow rate: 1 mQ/min; Detection: UV 254 nm
.

(c) N”-benzyloxycarbonyl-N”(N”
Benzyloxycarbonyl-shi-lysine)-L-lysine-P-nitrophenyl ester hydrochloride: N"-benzyloxycarbonyl-N-(N'"benzyloxycarbonyl-N/i-tert-butyloxycarbonyl- 10.25 g of the salt was prepared from 10.5 g of lysyl)-L-lysine-P-nitrophenyl ester according to the above process (,). M, P: 109~
113°C, TLCR, = 0.70 (II-BuOH-
^cOH-H, 0=4:1:l).

(d) N'-tert-butyloxycarbonyl-tri(N
= 1 benzyloxycarbonyl-1 lysine) -p-
Nitrophenyl ester: No-benzyloxycarbonyl-(Nε-tert-butyloxycarbonyl)-L-lysine 5.71 g (Is
mi, rimor).

Conversion to the mixed anhydride was carried out analogously to step (b) above using triethylamine and isobutyl chloroformate dissolved in acetonitrile. Then, No-benzyloxycarbonyl-Nε(N''-benzyloxycarbonyl-cylysyl)-L-lysine-P-nitrophenyl ester hydrochloride was added to this in the presence of triethylamine. Further, the above step (b) ) to obtain 13.9 g of product (yield 85.4%). This was further recrystallized from acetonitrile and ether. m, p,
: 145-150°C 0 analysis value: Content rate = 99.6%:
TLCR,=0.98(II-BuOH-AcOH-
)1. O=4:1:l):tlPLGtr
, = 18.2 min (K' = 8.1); Column: 0DS-Hyperzyl-6 (125X 4+m); Eluent: M
eOH-C1,CN-0,02M Na0AC! llW
Liquid (pH 4) = 40: 30: 30); Flow rate: 1 m
Q/min; Detection: UV 254nm.

(e) tri(N”-benzyloxycarbonyl-
Lysine)-p-nitrophenyl ester hydrochloric acid: 12.2 g of the ester produced in step (d) above was treated in the same manner as in step (a) above. This will reduce the salt to 12.
1 g (95.4%) was obtained. m, p, : 108
~125℃. TLCR, = 0.71 (II-BuOH:
AcOH: H, 0=4:1:l) −(f
) Poly-ε-N0-benzyloxycarbonyl-lysine: tri(No-benzyloxycarbonyl-lysine)-p-nitrophenyl ester hydrochloride 23 g (24
mmol) was condensed in 60 mQ of dimethyl sulfoxide in the presence of 4.5 taQ of triethylamine to obtain a gel. This polymer was poured into a 5% Na, Cos aqueous solution and separated.

Yield: 16.8g (89.6%1 Analysis value: Zco,
: 16.8% (calculated value), 16.3% (experimental value);
(rt sp)/c=31.92aj/g(c=4.9
5gIQ, dichloroacetic acid).

(g) Poly ε-L-lysine hydrobromide: the above step (
f) 16g of the protective polymer was added to 100ml of trifluoroacetic acid.
jl and further 4N HBr/AcOH10
Added 0weQ. This final product was precipitated using ether.

Yield f: 14.7g (91.6%, analysis value: Br:
37.7% (calculated value).

36.8% (experimental value); A 2 s 4 n s+
= 0, paper chromatography: vl 23x6
2. Eluent: n-BuOH-AcOH-Pyr.

-)1.0=30:6:24:20. Yield: 36.8
2% (calculated from the parent's child): 18.85% (
Calculated from free lysine MW=5,500-20
, 300° (h) Purification of the crude final product 250 g of the polymer obtained in step (g) above was dissolved in water and then chromatographed using a Sephadex G-50 (fine) column. Eluent: distilled water, 0.9% NaCl solution or 0.1N HCI solution. Detection of fraction (1 mQ): UV 220 nm.

(i) 2 g of purified poly-ε-lysine hydrobromide (obtained in step (g) above) of the crude final product was dissolved in 2 mQ of water and then 2 g of ethanol
I powered up the 0-kaku Q. This polymer was converted into diethyl ether 8
It was precipitated using 0mQ. After this was allowed to stand at 5°C for 24 hours, it was filtered, washed, and dried. Yield: 1.26 g (83%) (57.7% calculated from protected tripeptide starting material).

The resulting material was designated 5ZTP-14. M%l
=+2,700; (α)6”=+32-4@ (c
=2; n, o); Materials with polydispersity = 3.7° or less were prepared in a similar manner.

5ZTP-15: Mv =11-600 ±200
: (α)3' +31-9゜5ZTP-16:4=1
3.400f200: (a)♂'+32.6"5ZTP
−17: Mti =14.500f:200:Cαl
1g of the polymer obtained in Example 1 was added to 2M
NaCl solution 2M NaCl
The solution was further dialyzed against water. As a result of freeze-vacuum drying, 0.85 g of polymer was obtained.6 An ion exchange column (40 Q) was made from Dowex 1-OH anion exchange resin. Polymer produced as in Example 1. A solution of 1 g dissolved in 20 maQ water was subjected to chromatography at a flow rate of 1 cj/min. Both fractions were analyzed, collected, and lyophilized, resulting in the isolation of 0.65 g of the desired polymer.

10g of No.11 No-benzyloxycarbonyl-N[-tert-butyloxycarbonyl-〇-lysine-p-nitrophenyl ester was used as a starting material and treated in the same manner as in Example 1. As a result, 1.3 g of the target polymer was obtained - MV
=8900; (a) 3'=47-08' (c=0.
92:Wed).

According to the method of Example 1, No-benzyloxycarbonyl-(Na-tert-butyloxycarbonyl)-L-ornithine-p-nitrophenyl ester log to N''
7.9 g of -benzylcarbonyl-dis-ornithine-P-nitrophenyl ester hydrochloride was obtained. ■, p,: 8
At 7-90°C, this salt was treated with N as in step (1b) above.
“-benzyloxycarbonyl-(Na-tert-butyloxycarbonyl)-L-ornithine and N”
-Z-N'(No6-tert-butyloxycarbonyl-
A di-ornithine)-p-nitrophenyl ester dipeptide (12.1 g) was obtained. blasphemy, P
,: 133-136°C.

Repeat the above step (1a) to obtain N'-Z-Na-(N
'6-Z-L-ornithine)-L-ornithine-p-nitrophenyl ester hydrochloride was produced from the dipeptide (11.99 g: m, p,: 115-119°C), and from this the above step (lb) By coupling according to the method of Na-BOC-IJ (N"-
Z-L-ornithine)-p-nido 07:c-nyl ester (14.8 g, San, P.: 150-153°C) was obtained.

Furthermore, tri(N"-Z-L-ornithine)-p~nitrophenyl ester salt was produced by the method of step (1e) above. Layer, p: 115-130°C, this product 2
2 g was polycondensed in the same manner as in step (1f) above to obtain 15.4 g (87.6%) of δ-poly-N0-benzyloxycarbonyl-ornithine. From this product, 13.1 g (93%) of poly-δ-1-ornithine hydrobromide was obtained according to the method of the above step (1 g).

Using the method of Example 5, starting with 1 g of N''-benzyloxycarbonyl-Na-tert-butyloxycarbonyl-D-ornithine-p-nitrophenyl ester,
Poly-δ-rho-ornithine hydrobromide 1.2 g (89
%) was obtained.

Example 7 Poly-γ-L-α γ-diaminobutyric acid N″-benzyloxycarbonyl-Na-tert-butyloxycarbonyl-α-γ-diaminobutyric acid p-nitrophenyl ester in the same manner as in Example 1 9.8g to No-benzyloxycarbonyl-α,γ-diamino-butyric acid-p-nitrophenyl ester hydrochloride 6.9g
I got it. Peng, P.: At 67-70°C, this salt was treated in the same manner as in step (1b) above to give N''-benzyloxycarbonyl-Nγ-tert-butyloxycarbonyl-α,γ
- combined with diaminobutyric acid, N"-Z, Nγ-(N"
-Z-N'γ-tert-butyloxycarbonyl-α,γ-
Diamino)-butyric acid-p-nitrophenyl ester dipeptide (10.8 g, Sono, p.: 125-127°C) was obtained. The above step (1a) was repeated to obtain N"-Z-Nγ-(N"-Z-L-α, γ-diaminobutyryl)-L-α, γ-diaminobutyric acid-nitrophenyl ester hydrochloride 10 from this dipeptide. .35g was synthesized.

m, p,: i08-112°C. From this salt to the above step (1)
Coupling using method b) yielded 13.7 g of Nγ-BOC-tri(N"-Z-L-α, γ-diaminobutyric acid)-p-nitrophenyl ester. Peng, p.: 14
From this product, further tri(N"-Z-L-α, γ-diaminobutyric acid)-
10.5 g of p-nitrophenyl ester salt was obtained. ■,
p,: 105-110°C, followed by polycondensation in methyl sulfoxide according to the method of step (lf) above to obtain poly-
7.8 g (86.5%) of Qi NO-benzyloxycarbonyl-L-α,γ-diaminobutyric acid was obtained. This protected polymer was treated in the same manner as in the above step (1 g) to obtain 6.2 g (90.5 g) of poly γ-L-α, γ-diaminobutyric acid hydrobromide.

Using the method of Example 1, No-benzyloxycarbonyl-Nβ-tert-butyloxycarbonyl-L-α.

From 9.6 g of β-diaminopropionic acid p-nitrophenyl ester to No-benzyloxycarbonyl-
α.

7.1 g of β-diaminopropionic acid p-nitrophenyl ester hydrochloride was obtained at 0 m, p: 65-67°C,
This salt was combined with No-benzyloxycarbonyl-Nβ-tert-butyloxycarbonyl-α,β-diaminopropionic acid according to the method of step (1b) above, and N
'-Z-Nγ-(N''-Z-N'β-tert-butyloxycarbonyl)-α.

β-diaminopropionyl)-α,β-diaminopropionic acid-P-nitrophenyl ester dipeptide 11.
Obtained 0g. m, p,: 119-122°C. the above(
Repeat step 1a) to obtain N'-Z from this dipeptide.
-Nβ-(N'.

-Z-L-α, β-diaminobutyryl)-L-α, β-
10.85 g of diaminopropionic acid p-nitrophenyl ester hydrochloride was obtained. Yield: 10.85g, m
, p: 102-105°C. According to the method of step (1b) above, Nβ-BOC is obtained by coupling from this salt.
12.6 g of -tri(N''-Z-L-α,β-diaminopropionic acid)-p-nitrophenyl ester was obtained.
, P: 138-140°C. Furthermore, the above step (le)
Tori (N'-Z-L-α,
12.4 g of β-diaminopropionic acid)-p-nitrophenyl ester salt was obtained. n+, p,: 102-1
This compound was polycondensed in dimethyl sulfoxide at 06° C. according to the method of step (If) above to give 8.7 g of poly-β-N0-benzyloxycarbonyl-α,β-diaminopropionic acid (90, 2%). This protected polymer was treated in the same manner as in the above step (1g) to obtain poly-β-α.

β-diaminopropionic acid hydrobromide 5.6 g (88
, 7%).

500 mg of poly-L-lysine)IBr was dissolved in saline 10-Q. This injection solution was sterile filtered. The dosage is 1-100mg/kg/depending on the patient's condition.
used in days.

Tablets were made using the following ingredients.

Polyiso-L-lysine/HCI Lactose Saccharose Corn Starch Magnesium Stearate Alginate 10g 0g 6g 0g 2g 2g The above substances were mixed uniformly, and 100 tablets were press-molded from this mixture.

An aqueous solution was prepared from 5 g of photopolyiso-L-lysine using 100Ω of water. The pH of this solution was adjusted to 7.0-7.2 using NaOH solution. Then, 1 mQ of Tween 20 was added to this. This solution was sprayed over an area of 1 hectare to post-germinated plants to protect the plants.

Claims (14)

[Claims] (1) Compounds with the following general formula (I): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) (In the formula, R^4, R^5, R^6, R^7 and R^1^
0 is each independently a hydrogen atom, C_1_~_4 alkyl group; R^8 and R^9 are each a hydrogen atom; r is 10
an integer from 400 to 400; m is 0, 1, 2, or 3; n is 0) and salts, racemates, and optical isomers thereof. (2) R^4, R^5, R^6, R^7 and R^1^0
The compound of general formula (I) according to claim 1, wherein each is a hydrogen atom; r is an integer from 10 to 400; m is 0, 1, 2, or 3; and n is 0. (3) Polyisolysine having an average molecular weight of 5,500 to 25,600. (4) Polyisoornithine having an average molecular weight of 4,400 to 13,500. (5) Compounds of general formula (I) according to claim (1) (provided that R^4, R^5, R^6, R^7, R^8, R^9
, R^1^0 are each a hydrogen atom; r is 40 to 200
integer, m is 0, 1, 2 or 3; n is 0; amino group is L
1. An antitumor and antiviral drug composition, which contains a drug (type) as an active ingredient and is mixed with a drug carrier or a drug additive. (6) Active ingredients have an average molecular weight of 5,500 to 22,500
The pharmaceutical composition according to claim 5, which is polyisolysine. (7) Polyisodiaminocarboxylic acid of general formula (I) according to claim (1) (provided that R^4, R^5, R^6, R
^7, R^8, R^9, R^1^0 are the same or different and each means a hydrogen atom or an alkyl group of C_1_-_4; r is an integer of 10 to 400; m is 0 to 10
n is an integer of 0 to 10) and its salts, racemates, and optical isomers by coupling the corresponding monomers and further polycondensing the obtained oligomer; II) Monomer or oligomer: ▲ Formula,
There are chemical formulas, tables, etc.▼(II) (In the formula, q is an integer from 1 to 9; R^1 is an amino protecting group (
In the case of oligomers, all R^2 are the same); R^3 is an active ester protecting group for carboxyl groups suitable for protection as well as activation; R^4, R^5, R^6, R^ 7
, R^8, R^9, R^1^0, m and n are the same as above; however, in the case of oligomers, the substituents may be the same or different) are the monomers of the following general formula (III) : ▲There are mathematical formulas, chemical formulas, tables, etc.▼(III) (In the formula, R^2 is an amino protecting group that is different from R^1 and can be selectively removed separately from the R^1 group; X is a carboxyl activation group (however, -COX group is COOR^3
group); R^1, R^4, R^5,
R^6, R^7, R^8, R^9, R^1^0, m and n are the same as above), and the obtained compound of the following general formula (IV): ▲ Formula, There are chemical formulas, tables, etc. ▼ (IV) (In the formula, all substituents are the same as above; s is an integer from 2 to 10) If the R^2 protecting group is removed from and then s≦9, the obtained A compound of the general formula (III) is reacted with a monomer of the general formula (III) or polycondensed by a conventional method to obtain a compound of the general formula (III).
A process characterized in that, if the compound of I) is a free base, it is optionally converted into a salt, or if it is a salt, it is converted into a base or another salt. (8) Polyisodiaminocarboxylic acid of general formula (I) (
However, R^4, R^5, R^6, R^7, R^1^0 and r are the same as those in claim (1); m is 3; n is 0) and the method for producing its salt. , monomers or oligomers of general formula (II) (provided that R^1, R^3, R^4, R^
5, R^6, R^7, R^1^0 and q are in claim (1)
) is the monomer of general formula (III) (however, R^1,
R^2, R^4, R^5, R^6, R^7, R^1^0
and X are the same as in claim (1); m and n are the same as above)
The R^2 protecting group is removed from the resulting compound of general formula (IV) (all substituents are the same as above, s is the same as in claim (1)), and s≦9. 8. The method according to claim 7, wherein the compound obtained is optionally reacted with a monomer of general formula (III) or polycondensed by conventional means. (9) Polyisodiaminocarboxylic acid of general formula (I) according to claim (1) (provided that R^1, R^3, R^4, R
^5, R^6, R^7, R^1^0 and q are in claim (1
); m and n are the same as above) and a salt thereof, which comprises monomers or oligomers of general formula (II) (wherein R^1, R^3, R^4, ^5, R^6, R
^7, R^1^0 and q are the same as in claim (1); m and n are the same as above) to the monomer of general formula (III) (however,
R^1, R^2, R^4, R^5, R^6, R^7, R
^1^0 and X are the same as in claim (1); m and n are the same as above), and then the compound of general formula (IV) obtained (however, all substituents are the same as above; s is a claim (
(same as in 1)), and when s≦9, the resulting compound is optionally reacted with a compound of general formula (III) or polycondensed by conventional means. The manufacturing method according to claim (7), characterized in that: (10) A method for producing polyiso-L-lysine and its salt, comprising N^α-Z-L-lysine-p-nitrophenyl ester hydrochloride and N^α-Z-L-N^ε-BOC-
8. The production method according to claim 7, wherein L-lysine and isobutyl chloroformate are converted into a mixed anhydride and used as the starting material. (11) A method for producing polyiso-L-ornithine and its salt, comprising N^α-Z-L-ornithine-p-nitrophenyl ester hydrochloride and N^α-Z-N^β-BO
8. The production method according to claim 7, wherein C-L-ornithine is converted into a mixed anhydride together with isobutyl chloroformate and used as the starting material. (12) A method for producing polyiso-D-lysine and its salt, comprising N^α-Z-D-lysine-p-nitrophenyl ester hydrochloride and N^α-Z-N^ε-BOC-D-
8. The production method according to claim 7, wherein lysine and isobutyl chloroformate are converted into a mixed anhydride and used as the starting material. (13) A method for producing polyiso-D-ornithine and its salt, comprising N^α-Z-D-ornithine-p-nitrophenyl ester hydrochloride and NC^α-Z-N^β-B
8. The production method according to claim 7, characterized in that OC-D-ornithine converted into a mixed anhydride together with isobutyl chloroformate is used as a starting material. (14) A method for treating cancer, which comprises using an effective dose of a drug containing the compound according to claim (1) as an active ingredient.