DE2844501A1 - SALTS OF N-EPSILON -TRIMETHYLLYSINE, THE PROCESS FOR THEIR MANUFACTURING AND MEDICINAL PRODUCTS CONTAINING SUCH - Google Patents

Description

DR. STEPHAN G. BESZtDES PATENT ADVOCATE

APPROVED REPRESENTATIVE ALSO AT THE EUROPEAN PATENT OFFICE

PROFESSIONAL REPRESENTATIVE ALSO BEFORE THE EUROPEAN PATENT OFFICE

DACHAU NEAR MDNCHEN

posTFgcg 4 ^ 5 Q f

MDNCHENER STRASSE 8OA Federal Republic of Germany TELEPHONE: DACHAU 437f Post check account Munich CBLZ 700 100 80)

Account no. 1 368 71

Bank account no. 906 370 via the district and city savings bank Dachau-Indersdorf (BLZ 700 515 40) (VIA Bayerische Landesbank Girozentrale, Munich)

P 1 157

description

for patent application

JUDGE GEDEON VEGYESZETI GYAR R.T.

Budapest, Hungary

concerning

Salts of N-fc-trimethyllysine, method too their manufacture and medicinal products containing them

The invention relates to new N-fL-trimethyllysine salts » a process for their preparation and containing them Medicines, especially those against anemia or cytostatics with fewer harmful side effects.

The N- £ trimethyllysine has been found in numerous types of

909816/0982

Living things from the Actinomycetes to mammals and various plants in both bound and in found free state. The biological importance of this betaine-type amino acid and also that of related compounds, for example II-dimethyllysines and the methylated arginine, but could not be exactly clarified until today (Tyihak and coworkers: Life Science 20 [1977J, 385). Thus, N-t-trimethyllysine was found in more recently, for example, from the human placenta isolated (T. Tomita and K. Nakamura: Z. Physiol. Chem.

4-13) » ^{which indicates} a physiological or biochemical importance in the developing organs. The Kt-trimethyllysine reacts alkaline in the form of the base and thereby damages the cells. Since it is only bound by receptors in the form of its salts, it has no effect in vitro. The non-ferrous trimethyllysine was produced both per se and for various comparative experiments in the form of various salts thereof. One of the best-studied salts is NE trimethyl lysine dioxalate (T. Takemoto and co-workers: Yakugaku Zasshi 84 [1964], 1 176; Japanese patents 24 790 ('65), 09 529 ('68) and 28 092 ('68); French patent specification 1 442 318; M. Takehara and colleagues: Nippon Kagaku Zasshi 1969, 101), which is a stable salt, but is not suitable for pharmaceutical use. The more thoroughly investigated salts also include N- £ -trimethyllysine- monohydrochloride and dihydrochloride (British patent 865 157; French patent 1 442 318; J. Puskäs and E. Tyihak: Periodica Polytechnica 1j £ [1969]] »261) ; JH Seely and KL Benoiton: CaiSi. J. Biochem. [19703, 1122; RA Cox and CL Hoppel: Biochem. J. [19733 j 1 083). However, these salts are highly hygroscopic, which makes their storage and dosage problematic. The salts of N- £.-Trimethyl ^x lysines were further comprising Nt-trimethyllysine hydrobromide (French Patent

909816 / 09S2

1 442 318), the NE trimethyl lysine citrate (H. Ozawa and coworkers: Yakugaku ZassM &] _ [1967], 935), the NL-trimethyl lysine di- (p-hydroxyazobenzene-p'-sulfonate) ■ JR. T. Markiw: Biochem. Med. 13 £ 1975] ι 23 '; I. Kakimoto and S. Akazawa: J. Biol. Chem. 245 [197], 5,751; T. Tomita and K. Nakamura: Z. Physiol. Chem. 358 [1977], 413j, the li-L-trimethyl lysine nicotinate (French patent Λ 442 318) and the Nt-trimethyl lysine N-acetyl-L-amino-succinate (French patent 1 442 318) ". However, none of these salts meet the requirements placed on stability and pharmaceutical usability.

The invention is based on the object of providing superior pharmaceutically usable new N- £. -Trimethyllysine salts which stable, i.e. not hygroscopic, easy to dose and store, a precisely defined composition have and in aqueous solution a pH value that is equal to or equal to the pH value of the physiological saline solution to provide a method for producing the same and medicaments containing the same.

It has now surprisingly been found that the salts of N-E-irimethyllysine with 1 or 2 molecules Glutamic acid, fumaric acid and aspartic acid meet these requirements.

The invention therefore relates to the salts of N- ε-trimethyllysine with 1 or 2 molecules of glutamic acid, Fumaric acid or aspartic acid.

The N-t-trimethyllysine salts according to the invention can present both in racemic and in optically active form, but work based on the tests carried out the levorotatory forms, i.e. the N- £ -trimethyl-L- -lysine salts, most advantageous, which is why they are preferred,

909816/0952 " ⁴ "

A particularly preferred compound according to the invention is N- £ -trimethyl-L-lysine monoglutamate.

Further preferred compounds according to the invention are N-t-trimethyl-L-lysine difumarate and li-f-trimethyl-L-lysine - slide paraginate.

The invention also relates to a method for {. -j Production of the compounds according to the invention, which da-f. fl \ is characterized by that \ ^r M- £ -trimethyllysine in aqueous \ '.-_% ger solution with glutamic acid, fumaric acid or aspartic; A'ü

i -'- O acid is implemented. j 3 "

It is expedient to proceed in such a way that the aqueous solutions of the starting materials are combined with one another be, the reaction mixture is evaporated after the end of the reaction and the residue, if necessary after recrystallization, is dried.

The amount of glutamic acid, fumaric acid or aspartic acid used as the starting material depends of course, it depends on whether a salt with 1 or 2 molecules of acid is to be produced.

Furthermore, pharmaceuticals according to the invention, which 1 or more of the compounds according to the invention as an active ingredient or Active ingredients, optionally together with 1 or contain more other pharmacologically active compounds and / or conventional carriers and / or auxiliaries, provided. As already mentioned, the compounds according to the invention have valuable pharmacological effects, especially against anemia, especially tumor anemia.

The pharmacological action of the compounds according to the invention was demonstrated by the following experiments.

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First, the effect of N-ε-trimethyl-L-lysine-monoglutamate on the blood count of mice infected with the tumor NK / Ly-ascites was investigated. ^> O mice of the strain CFLP O (weight: 20 to 22 g, age: 8 weeks) were infected intraperitoneally with the tumor NK / Ly-ascites (5 × 1 Cr cells per animal). Of the 50 animals served as Blindversuchs- 25 or control animals, the remaining 25 were ten-2 on, th-5, 9- "t ^s i 11-th, 13 th and 19 th day after infection (at the 19 th On the 2nd, 5th, 8th, 10th, 12th, 13th day, only the 5 surviving animals were treated intraperitoneally with 100 mg / kg of LV-trimethyl-L-lysine monoglutamate The blood count was examined on the th, 14th and 20th day after the infection, and the results are shown in Table 1 below.

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- 6 table

co ο co

Day of blood count
investigation Kote Blutk <
Xl
Million"
at
treated
Animals jrperchen
a
3n / mm
the
Blind test
animals White blood kos
in
'fausendei
at
treated
Animals rperchen
i / mm
the
Blind test
animals 2 8.66 7.06 6.446 7,800 5 7.2 6.95 8.732 8,466 8th 8.2 7.8 7,200 7.733 10 7.2 6.4 8.520 8,900 12th 7 ^ ^, 7 10,800 6,500 13th 6.6 4.9 8.260 6,700 14th 3 ^ 4.5 9,000 7.700 20th 5.0 2.1 7.133 5.100

CT) CD

From the data in Table 1 above it can be seen that the N-8-tri-diethyl-L-lysine monoglutaniate which changes in the course of the growth of the NK / Ly-ascites tumor Anemia decreased considerably. The intraperitoneal LD ^ Q value of this compound in mice is above 15 ° O mg / kg.

Furthermore, the incorporation of thymidine (- " ¹ H Täü) into the thymus, the bone marrow, the spleen and the mucous membrane of the small intestine of 12-week-old healthy female mice of the strain DBA / 2 as well as in vivo proliferating tumor cells of the type NK / Ly -Ascites. 5 animals were used per group and time point. The experiments were carried out in 4 repetitions, the NS-tri-diethyl-L-lysine- monoglutamate being administered intraperitoneally at a dose of 100 mg / kg. It was found that that 24 hours after the treatment with the Ni-trimethyl-L-lysine monoglutamate the incorporation of the thymidine in the thymus by 85 ± 7%, in the spleen by 75 ± 8% and in the tumor cells of the type XK / Ly-ascites by 94 ± 11% was increased. The cells of the bone marrow showed in the 10th and 48th hour also an increased (by 57-4% and 72 ± 5%) incorporation of thymidine. Ly-ascites, the incorporation of the thymidine in the 48th hour was even more important (at 12 4 ± 14%) reinforced. At peroral administration was 32 hours after the treatment, an increase of the incorporation of the Thymidines in the mucosa of the small intestine by 50 - to watch 9%. This shows that the cells have started deoxyribonucleic acid synthesis and have entered the active phase.

The transforming effect exerted on the lymphocytes was also examined in human cultures. From the peripheral Blood from healthy blood donors were isolated from lymphocytes using the Picoll- -Uromiro gradient. These were in a cell count of 5 x ICK / cnr in an autologous serum

909816/0952 " ⁸ "

containing medium of the type Parker 199 suspended and incubated at 37 ° C for 96 hours. Some of the samples were treated with 100 lg / cn * K - ^ - trimethyl-L-lysine dihydrochloride and another part with 100 ug / cnK Nt-trimethyl-L-lysine- monogluteEat and the remaining samples were used as blank test- or control samples. The converting activity was determined by the incorporation of the Kaß ThyEidin - determined .. The treatment in question with the thymidine was made in a dose of 11 uCi / cnr 3 hours before termination of incubation ⁽⁵ H TDB). The cultivation of the cultures was stopped 48, 72 and 96 hours after the start, respectively. The activity bound to deoxyribonucleic acid was determined by liquid scintillation after extraction with boiling superchloric acid. The results obtained are shown in Table 2 below.

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Tabel

ca O co oo

Time Uptake of thymidine PH TdE] \ (cpi ΙΤ-ε-ΪΓχίΐΐΘΐΙ ^ Ι-ΐ - '^ Βάη- N- £ -trimethyl-L-lysine- α) - ^ \ ~ · '- ^w - ^ in in treatment dihydrochloride -monoglutamate hours with 146 594 - 756 no means 167 413 (Blind test) 295 3 010 48 303 2,427 123 254 1 413 231 2,825 72 330 2,878 159 301 3 659 96 271

subsequent
[geä

- ^ This is of fundamental importance, since the relatively low effectiveness of numerous cytostatics is due to the fact that the division of the tumor cells is not sufficiently intense, so if the multiplication of the tumor cells can be increased by division, the cytostatics administered at the same time can have a much more intense effect . * * ^

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- AO

Table 2 above clearly shows that thymidine uptake and thus that bound to deoxyribonucleic acid Activity in treatment with N-JL-trimethyl-L- -lysine monoglutamate was far higher than that of the treatment with itf-t-trimethyl-L-lysine dihydrochloride.

The fact that the resting cells enter the active phase is proven by the fact that they begin to multiply. Therefore, the development of the total number of tumor lines was also examined.

20 female mice of the Ci ¹ LP strain were vaccinated with the M / Ly-ascites tumor. 24 hours later, 10 animals were administered 100 mg / kg K-6-trimethyl-L-lysine monoglutamate daily for 8 days, while the remaining 10 animals served as blind test animals or control animals. 9 days after vaccination, the mice were sacrificed and the weight of the animals and the total number of tumor lines were determined. It was found that the total number of tumor lines in the treated animals was 35% higher than in the blind test animals. \ ~ - ~?

subsequently changed

Close!:? cn the effect of N-t-trimethyl-L-lysine -monoglutamate on humoral and cellular immune response studied by mice. L-6-phenyl-2,3,5,6-tetrahydroimidazo [2,1-b] thiazole was used as a comparison substance {LevamisoleJ. The investigations were carried out on groups of 10 2-month-old female mice of the strain DBA / 2, with the red blood cells of the sheep intraperitoneally as antigen were administered. As a positive control, E. coli endotoxin and a 1-time treatment with 2 to 5 mg / kg L-6-phenyl-2,3,5,6-tetrahydj? Oimidazo £ 2,1-bJthiazole used. The li-t-trimethyl-L-lysine monoglutamate was as a pretreatment for 2 weeks at a dose of 5 x 100 mg / kg intraperitoneally before the administration of the Antigen administered. The hemagglutinin titer was

- 11 -

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Determined using a microtitrator according to Takacs. It was found that pretreatment with N-fL-trimethyl-L- lysine-monoglutaiTiat brought about an equally large increase in titer as that with the _{L-6-phenyl-2,3-5} 5J6 tetrahydroimiaazo [2,1-b2thiazol. The latter, however, has various side effects, for example a harmful effect on the Kagen-Barm-Trakt, while the Ν-Ε, -trimethyl-L-lysine- -eonoglutamate has no side effects because it is the salt of a substance which is also in the organism occurs is.

To study the cellular immune response was applied the experimental model described by Legrange and co-workers. Its essence is that after several times Antigen administration (red blood cells of the sheep) the sole thickness of the mice grows. The measure of this growth will be generally reduced by the agents that stimulate the humoral immune response in relation to the control value, after a few days, however, increased in relation to this already reduced control value. This appearance tr: Jt even with pretreatment with N-E-trimethyl-L-lysine monoglutamate (5 x 100 mg / kg within 2 weeks).

Based on the above test results, the advantages of N-E-trimethyl-L-lysine monoglutamate can be summarized as follows will:

a) Even when administered alone, it improves in tumor anemia, but also in Anemia of other origin the blood count.

b) Its immune-stimulating effect can be used in tumor diseases, but also in others with a Decrease in humoral immunity related cases are well utilized.

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c) Since it makes numerous cells sensitive to cytostatics, it is in combination with Cytostatica are well applicable in the therapy of tumor diseases.

d) As a result of its effect on the blood count, uxL of the mucous membrane entering the intestine the growth (proliferation) increasing effect it reduces when administered at a suitable time the toxic effects of the Gytostatica.

According to an advantageous embodiment, the medicaments according to the invention contain 1 or more cytostatic active ingredients as other pharmacologically active compound (s). They can expediently be used as a cytostatic agent, a cytostatic agent or cytostatic agent Ii, N-bis- (2-chloroethyl) -tetrahydro- -2H-1,3,2-oxazaphosphorin-2-amine-2-oxide (cyclophosphamide), 1,4 ~ Di- (2-methylsulfonyloxyäthylamino) -1,4-dideoxyerythritol dimethylsulfonate (Ritosulfanum), vincristine, vinblastine, leuurosine and / or B-jormylleurosine, optionally in salt form.

In the above-mentioned medicaments according to the invention, the quantitative ratio of the N- ε-trimethyllysine salt or the NE-trimethyllysine salts to the cytostatic agent or to the cytostatic agent, depending on the usual dosage of the cytostatic active ingredient or the cytostatic active ingredients, is expediently 1: 100 to 100: 1, in particular 1 : 1 to 100:

These medicaments according to the invention bring the big one technical port step with it that with them the toxic side effects of the known cytostatic agents are reduced to a considerable extent.

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This significantly reduces the adverse effects on the blood count. The blood counts were done carried out as follows:

A) There were 6 8-week-old healthy female mice of the strain CFLP (MTI) with body weights of 20 to 22 g n.a. with a single intraperitoneal dose of 30 mg / kg 1,4-di- (2-methylsulfonyloxyethylamino) -1 , 4-dideoxyerythritol dimethyl sulfonate treated. In addition, 6 such animals were treated with a medicament according to the invention which, in addition to 1,4-di- (2-methylsulfonyloxyäthy1amino) -1,4-dideoxyerythritol dimethyl sulfonate from K-6-trimethyl-L-lysine monoglutamate in the weight ratio of the former to the latter of 30: 100, treated at a dose of 130 mg / kg intraperitoneally. On the 2nd, 4th, '/' - th, 10th and 17th day after the treatment, qualitative and quantitative blood count determinations were carried out. Blood was drawn from 3 animals in each group, always at 3 p.m. on an empty stomach. The results are compiled in Table 3 below.

B) 5 8-week-old healthy female mice of strain Ci ¹ LP (LATI) with body weights of 20 to 22 g were given once with an intraperitoneal dose of 300 mg / kg N, N-bis- (2-chloroethyl) - tetrahydro-2H-1,3,2-oxazaphosphorin-2-amine-2-oxide. In addition, 5 such mice were treated with a medicament according to the invention which, in addition to the N, N-bis (2-chloroethyl) tetrahydro-2H-1,3,2- -o: mzaphosphorin-2-amine-2-oxide from HC- Trimethyl- L-lysine monoglutamate in the weight ratio of the former to the latter of 3 J 1 existed, in

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at a dose of 400 mg / kg intraperitoneally. On the 4th and 10th days after the treatment, the quantitative blood count became from 3 animals each determined. These results are too compiled in the following table 3.

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-Αϊ

Table 3

treatment Day animals Red blood White blood with Kr. corpuscles corpuscles in
Millions / min in
Thousands / mm ^ 1 7.0 3.6Ou 2 2 6.8 3,200 .3 7.2 3,400 Dutchsdinitt 7.0 3,400 1 4-Tue- (2- 1 5.6 3,400 -methyl-
sulfonyl
oxyethyl 4th 2
3
average 6.8
6.4
6.3 6,200
5,200
4-, 933 amino) - 1 4.6 5,600 -1,4-di- 7th 2 4.0 2,600 deoxy 3 6.4 4,000 erythritol Dtoxhschnitt 5.0 4.066 -dimethyl-
sulfonate
[[alone] 10 1
2
3 • 7.4-
2.8
5.6 11,800
10,600
6,600 average 5.2 9,600 1 6.7 5,200 17th 2 6.0 5, ooo 3 6.2 5.100 TTnrnhRf + TrTThh 6.3 5.100

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Table 3 continued

treatment
with Day animals
Kr. Red blood
corpuscles
in
II i 11 i onen / imir White blood
corpuscles
in
Thousand- d / ™ {-jnethyl-
sulfonyl
oxyethyl
ainino) -
-1,4-di-
deoxy
erythritol
-dimethyl-
sulfonate +
+ N t
-L-lysine-
-mono-
glutamate 2 1
2
3
Dor chschriitfc 7.6
7.0
7.6
7.4 2,600
2,600
2,600
2,600 4th 1
2
3
E-headcut 7.6
7.0
6.4
7.0 5,600
6,200
7,500
6.366 7th 1
2
3 6.6
7.6
7.0
7.0 5,000
5,700
6,400
5,700 10 5.8
5.6
5.2 11,800
8,000
9,000
9,600 17th 1
2
3 10.0
9.6
9.2
9.6 10,400
10,800
10,600
10,600 1
2
3

909816 / 09S2

Port,

the table

Treatment; Day animals Red blood White blood idLt Kr. corpuscles corpuscles in in Fiilliorien / FJTi A thousand and one rair K, K-bis- (2- 1 5.4 8,600 -Chlorine- 2 6.0 3,500 ethyl)- 3 7.2 7,200 -1 etr ί ύ Jj) dro- Daithschoitb 6.2 6.433 -2H-1,3,2-
-OX & Zc-- phogiiorin- 1 4.8 18,000 -2-amine- 10 2 4.4 16,000 -2-oxide 3 9.2 14,000 alone] D ^ tfjsdxätfc 6.13 16,000 -Chlorine- ethyl)- -tetarahydro- 1 7.8 6,600 -2H-1,3,2- 4th 2 8.2 4,400 -oxaza- 3 7.0 9,000 phosphorine DjTchscbnitt 7.6 6.333 -2-amine- 1 6.8 16,000 -2-oxide. + 2 9.6 16,000 + N-t- 10 3 5.6 19,100 —Tdmeoiyl— IXuxi section 7.3 17,200 -L-lysine- -mono- glutamate

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From the data in Table 3 above, it can be seen that in the treatment with the medicament according to the invention Also containing N-S-trimethyl-L-lysine monoglutamate in addition to the 1,4-di- (2-methylsulfonyloxyäthyl..iiiino) - -1,4-dideoxyerythritol diicethylsulphonate the blood cells Both the red and the white blood cells were much more favorable than those of the treatment with 1,4 ~ di- (2-methylsulfonyloxyäthylamino) -1,4-dideoxyerythritol ~ dimethylsulfor.-at alone. The ratio of the lymphocytes to the granulocytes was determined by combining the 1,4-di- (2-methylsulfonyloxyäthylaEino) - -1,4-dideoxyerythritol dimethylsulfonates with the N- £ - -Trimethyl-L-lysine-monoglutamate practically not influenced. Furthermore, it can be seen that in the case of the medicament according to the invention with a content of N-E-trimethyl-L-lysine -monoglutamate in addition to the N, N-bis- (2-chloroethyl) -tetrahydro- -2H-1,3,2-oxazaphosphorin-2-amine-2-oxide treated animals the number of red blood cells was higher than in those with the üi, H-bis- (2-chloroethyl) -tetrahydro-2H-1,3,2-oxazaphosphorin-2-amine-2-oxide treated alone.

Bone marrow damage plays a role in the toxic, often fatal effects of the cytostatic agents essential role. In the following experiments it was therefore investigated whether and to what extent the toxicity or the LDj-Q value of the cytostatic agents in combination with N-t-trimethyl-L-lysine monoglutamate in the inventive Medicines is influenced in a favorable direction. In the comparative experiments carried out, the toxicity of 3 different cytostatic agents, namely N, N-bis- - (2-chloroethyl) tetrahydro-2H-1,3,2-oxazaphosphorin-2-amine- -2-oxide, vincristine and 1,4-di- (2-methylsulfonyloxyäthylamino) -1,4-dideoxyerythritol dimothylsulfonate, with that of drugs according to the invention, which combinations

- 19 909816 / 09S2

this gown contained N-E-Trifflethyl-L-lysine-monoglutamate, compared.

The tests were carried out on female cows from Stanm CBA with body weights of £ 5 g with groups of 5 animals each. In individual groups, the animals were treated with a 1 - ;;, a] igen in.ti-aperitor.c-alen dose of Ai- ^ jeiratteln of the Zuc & Tj.en-EttLungen "given in Table 4 below.

^S the number of surviving animals was registered in the first 10 days and at 30 ~ "ten days after treatment. The results are also summarized in Table 4 below.

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- 20 Table 4

to ο co

cn

co cn

Ver
search
No. Drug composition Number of surviving animals
Day after treatment
1 2 3 4 5 6 7 8 9 10 30 , 1 600 mg / kg N, N-bis- (2-chloroethyl) -tetrahydro-
-2H-1,3,2-oxazaphosphorin-2-amine-2-oxide +
+ 100 mg / kg Ν-ε-trimethyl-L-lysine monoglutamate 555555555 5 3 2 5 mg / kg vincristine +
+ 100 mg / kg NE-trimethyl-L-lysine-monoglutamate 5555555 ^ 4 3 3 3 50 mg / kg 1,4-di- (2-methylsulfonyloxyethyl-
amino) -1, 4-dideoxyerythritol dimethyl sulfonate +
+ 100 mg / kg Nt-trimethyl-L-lysine monoglutamate 555555555 5 4 4th 675 mg / kg N, W-bis- (2-chloroethyl) -tetrahydro-
-2H-1,3,2-oxazaphosphorin-2-amirL-2-oxide
[Comparative material] 000 000 000 0 0 5 5 mg / kg vincristine j reference material] 554444443 3 3 6th • 45 mg / kg 1,4-di- (2-methylsulfonyloxyethyl-
amino) -1,4-dideoxyerythritol dimethyl sulfonate
j [comparison material] 555544322 2 2

OsJ

OO

cn

From the data in Table M above, it can be seen that the extent and the rate of death of the animals in the groups treated with the medicaments according to the invention show a much more favorable I Id than in those treated only with the white ones cytostatic agents (reference materials) treated animals. The medicaments according to the invention showed a clearly renewable protective effect against the toxic effects caused by the cytostatic agents.

The daily dose of the L-C-trimethyl-L-lysine salt or the Κ--trimethyl-L-lysine salts of the medicaments according to the invention is expediently 5 to 50 mg / kg, preferably 20 to 40 mg / kg. The useful daily dose of the cytostatic active ingredient or the cytostatic active ingredients of the medicaments according to the invention is dependent on the type of cytostatic agent used or the cytostatic agent used. For example, in the case of 1,4-di- (2-methylsulfonyloxyethylamino) -1,4-dideoxyerythritol dimethylsulfonate, 20 to 40 mg / kg, in the case of N, N-bis (2-chloroethyl) tetrahydro -2H-1,3,2-oxaza phosphorin-2-amine-2-oxide 10 to 100 mg / kg and in the case of vincristine 0.1 to 10 mg / kg, preferably 1 to 5 mg / kg, is expedient. The amount of the doses to be administered in specific cases is determined taking into account the type of treatment, the patient's condition and the severity of the disease. The individual medicaments according to the invention are expediently produced in two different doses: in medium-strength or strong (forte) individual doses with a correspondingly proportioned content of cytostatic active ingredients.

The invention is explained in more detail by means of the following examples.

When making the connections, the

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- -32 -

Always use a Hotavapor device (Büchi) for evaporation. the Melting point was determined using a Tottoli apparatus (Büchi) carried out. The thin-layer chroniatograiDme were added to fixion layers. A Solution of 50 g citric acid, 50 g sodium hydroxide and

Z Z

7 cin ^ concentrated hydrochloric acid in 500 cm / distilled Water used. The chro: n ^ to £; i'avrae were made with a

ζ ζ

Solution of 80 cm ^y mth of oil, 20 cz. You vinegar, G, 2g Ii in-

hydrin and 0.1 g copper (JI) sulfate.

Example 1 Ή-Ζ -trimethyl-L-lysine-monoglut amat

50.0 g (0.265 mol) of chromatographically uniform K- ε-trimethyl-L-lysine were dissolved in 500 curd of distilled water. The solution was warmed to 50 ° C. and combined with a solution of 3910 g (0.265 mol) of L-glutamic acid in 500 ml of distilled water. The solution obtained was stirred or shaken for minutes at 50 ° C and then 5% strongly adsorbing activated carbon (Norit activated carbon) obtained ^{from plant matter was added.} The solution was stirred or shaken for a few minutes, then filtered and evaporated to dryness under reduced pressure. On evaporation, the temperature of the solution was not allowed to rise about 5O ⁰ C. The crystalline residue obtained was dried under vacuum over phosphorus pentoxide to constant weight. 95 to 98 g of non-ferrous trimethyl-L-lysine-mono-glutamine with a melting point of 104 to 107 ° C. and a [<*] _{jp_yert} of + 5.15 ° (c = 1, water) were obtained.

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Imino acid analysis:

calculated: glutamic acid = 1 hol,

N-i-tri-ethyl-L-lysine = 1 KoI;

found: glutamic acid = 1.0 mol,

N-i-trimethyl-L-lysine = 1.13 moles

door C ₁₄ H ₃₀ O ₆ N ₃ . 2 H ₂ O

calculated: C = 45.2%, N = 11.3%; found: C = 45.6%, Ή = 11.2%.

Example 2 li-L-trimethyl-L-lysine difumarate

There were 7 »0 g (0.37 mol) chromatography sch unit-

7. Liches Nc-trimethyl-L-lysine dissolved in 70 cnr distilled water. A boiling solution of 99.5 g (0.85 mol) of fumaric acid in 4,000 cubic centimeters of ethanol was added to the solution with stirring or shaking. The slowly cooled mixture was kept at 0 ° C. for 48 hours. The precipitated crystals were filtered off, washed first with ethanol and then with ether and finally dried. 108 g (69% of theory) of N-6-trimethyl-L-lysine difumarate were thus obtained. This was recrystallized from ethanol containing 3% water. The product had a melting point of 147 to 149 ° C (with decomposition), an R ^ value of 0.1 and a [ρί] π value of + 6.0 ° (c = 2, water).

Amino acid analysis:

calculated: N- ε-trimethyl-L-lysine = 1 mol found: N-6-T: rimethyl-L-lysine = 1.0 mol.

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Example 3 , Kt-trimethyl-L-lysine diasparaginate

There were 5O ₁ O g (0.265 mol) chroKatographisch uniform Iv-t-Trinethyl-L-lysan dissolved in 500 cnr distilled water at a temperature of 50 ° C. This solution was added to a 50 ° C. solution of 70.1 g (0.53 mol)! -Aspartic acid in 500 cnr distilled water. The solution was stirred or shaken for a few minutes and then 5 g of the activated carbon used in Example 1 were added. After filtering, the solution was evaporated under reduced pressure. The oily residue was solidified by treating with dry ethanol. Thus 60.0 g of HE trimethyl-L-lysine diasparaginate with a melting point of 14-2 to 153 ° C. were obtained. (The substance was very hygroscopic).

Example J \ _

There were 500 mg ΙΓ-ζ-trimethyl-L-lysine monoglutamate, 500 mg li, K'-bis- (2-chloroethyl) -tetrahydro-2H-1,3,2-oxazaphosphorin-2-amine-2-oxide and 200 mg of protein-free gelatin dissolved in distilled water suitable for injections, the volume of the solution was adjusted to 35 »0 cm, and the solution was freed from bacteria by filtration under aseptic conditions. Of the sterile solution is filled cans were cur from each 3 '5 in sterile Ί0 cnr Pläschchen, the open Pläschchen were 20 hours maintained at -6O ⁰ C and then lyophilized at a pressure of 25 mm Hg at -45 ° C and + 20 ^° C. post-dried for 10 hours.

Then the vials were fitted with sterile rubber stoppers and aluminum caps closed. The on this

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Dry ampoules produced in this way contained 50 mg of non-ferrous trimethyl-L-lysine monoglutamate, 5> O mg of N, N-bis- (2-chloroethyl) -tetrahydro-2H-1,3 ₅ 2-oxazaphosphorin-2- - per ampoule amine-2-oxide xznd 20 mg gelatin in lyophilized state. Prior to administration of the injection, the contents of the vial were dissolved in 10 cnr distilled water suitable for injections or in isotonic saline or glucose solution.

Example 5

There were 500 mg of IJ-E-trimethyl-L-lysine monoglutamate, 300 mg of 1,4-di- (2-methylsulfonyloxyäthylamino) -1,4-dideoxyerythritol dimethylsulfonate and 400 mg of gelatin dissolved in distilled water suitable for injections and the The solution was adjusted to a volume of 10.0 cm and treated further in the manner described in Example 4, with the difference that in each case 1 cm of sterile solution was filled into the ampoules. The finished preparation contained 50 mg IT-E-trimethyl-L-lysine monoglutamate, 30 mg 1,4-di- (2-methylsulfonyloxyethylamino) -1,4-dideoxyerythritol dimethylsulfonate and 40 mg gelatin in lyophilized state per ampoule.

Example 6

500 mg of 2i- ε-trimethyl-L-lysine monoglutamate, 5 mg of vincristine sulfate and 200 mg of gelatin were dissolved in distilled water suitable for injections and the solution was adjusted to a volume of 10 ^ 0 cnr and that described in Example 5 Way further worked up. The finished preparation contained 50 mg If-C-trimethyl-L-lysine monoglutamate, 0.5 mg vincristine sulfate and 20 mg gelatin in lyophilized state per dry ampoule.

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Example 7

There were 500 Eg JJ-i-trimethyl-L-lysine-raonoglutamat, 5 mg vincristine sulfate, 80 mg polyvinylpyrrolidone and 36.5 Eg Mannitol dissolved in distilled water suitable for injections, the volume of the solution was adjusted to 10.0 enr and the solution was further treated in the manner described in Example 5. The preparation obtained contained 50 mg N-t-trimethyl-L-lysine monoglutamate per dry ampoule. 0.5 mg vincristine sulfate, 8 mg polyvinylpyrrolidone and 36.5 mg of mannitol in a lyophilized state.

Example 8

There were 50 mg of NE trimethyl-L-lysine monoglutamate. 50 mg N, N-bis (2-chloroethyl) -tetrahydro-2H-1 _i 3,2-oxazaphosphorin-2-amine-2-oxide, i mg colloidal silica, Λ mg magnesium stearate, 2 mg talc and 56 mg potato starch mixed homogeneously in the form of a dry powder and the 160 mg powder mixture obtained was filled into a hard gelatin capsule. The preparation obtained was suitable for oral administration.

Claims 909816/0952

Claims (1)

2.) ch "; J-ilch 6.) 7.) 8th.) Salts of N-t-trimethyllysine with 1 or 2 molecules Glutamic acid, fumaric acid or A sp ar ag in acid. IT-f-trimethyl-L-lysine pionoglutamate. Ή-ξ. -Trimethyl-L-lysine difumarate. li-E-trimethyl-L-lysine diasparaginate. experienced for the preparation of the compounds according to nspruch 1 to 4, characterized in that one N- £ -trimethyllysine in aqueous solution with glutamic acid, Converts fumaric acid or aspartic acid. Medicament, characterized in that it contains 1 or more compounds according to Claims 1 to 4 as an active ingredient or active ingredients, optionally together with Ί or more others pharmacologically active compounds and / or customary carriers and / or auxiliaries. Medicament according to Claim 6, characterized in that it is used as other pharmacologically active compounds) Contain 1 or more cytostatic agents. Medicament according to claim 6 or 7 »characterized in that that they are used as a cytostatic agent or a cytostatic agent N, N-bis (2-chloroethyl) tetrahydro-2H-1,3,2-oxazaphosphorine-2-amine-2-oxide contain. 909816/0952 2644501 9.) Medicament according to claim. 6 to S, characterized in that that they are used as a cytostatic agent or a cytostatic agent 1,4-di- (2-methylsulfonyloxyäthylamino) -1,4-- -dideoxyerythritol dimethyl sulfonate contain. 10.) Medicament according to claim 6 to 9, characterized in that that they are used as a cytostatic agent, a cytostatic agent, respectively cytostatic agents vincristine, vinblastine, leurosine and / or N-formylleurosine, if appropriate in courtship form. 11.) Medicament according to claim 6 to 10, characterized in that the quantitative ratio of the υ-έ, -trimethyllysine salt or the N-t-trimethyllysine salts for cytostatic agent or to the cytostatic agents 1: 100 to 100: 1, in particular 1: 1 to 100: 1, is. 909816/09 5 2