CA2048627A1 - 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane derivatives, pharmaceutical compositions containing them and their use for the removal of toxic metal ions and radioactive isotopes from the living organism - Google Patents

Abstract

1,4,10,13-TETRAOXA-7,16-DIAZACYCLOOCTADECANE DERIVATIVES PHARMA-CEUTICAL COMPOSITIONS CONTAINING THEM AND THEIR USE FOR THE
DECORPORATION OF METAL IONS, MAINLY RADIOACTIVE ISOTOPES DAMAGING
THE LIVING ORGANISM

Abstract The invention relates to partially novel metal complexes, salts and double salts of 1,4,10,13-tetraoxa-7,16-diazacycloocta-decane compounds of the formula (I), (I) wherein Q1 and Q2 mean hydrogen or a group of the formula (III), (III) in the groups of the formula (III) the R subtituents independently represent hydroyen, a C1_5 straight or branched chain alkyl group, a C2-5 straight or branched chain alkenyl group, phenyl or phenyl-C1-5alkyl group, the two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), C1-5alkyl, C1-5alk-oxy, cyano or nitro group(s), with the proviso that at least one of Q1 and Q2 is other than hydrogen;
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is 0 or 1;

M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of M, N or Me, respectively;
s and r are, independently from each other, O, 1, 2, 3 or with the proviso that (i) r, s and q cannot simultaneously be 0; and (ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2, as well as pharmaceutical compositions containing these com-pounds.
The compounds and compositions according to the invention can be used for the decorporation of metal ions, mainly radio-active isotopes such as radioactive strontium or cerium being harmful to the living organism. The active agents according to the invention exert their effects either in intravenous or local, transdermal or rectal route of administration.

Description

20~862-7 1,4,10,13-TETRAOVA-7,16-DIAZACYCLOOCTADECANE DERIVATIVES, PHARMA-CEUTICAL COMPOSITIONS CONTAINING THEM AND THEIR USE FOR THE
REMOVAL OF TOXIC METAL IONS AND RADIOACTIVE ISOTOPES FROM THE
LIVING ORGANISM

The~invention relates to partially novel 1,4,10,13-tetraoxa--7,16-diazacyclooctadecane derivatives and the use of such com-pounds for the decorporation of metal ions, mainly radioactive isotopes damaging the living organism. More particularly, the invention relates to metal complexes, salts and double salts of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane derivatives of the formula (I) Mn~ Meq 3 - ~ I Ns tl~
wherein Ql and Q2 mean hydrogen or a aroup of the formula (III), COO ~
I

--C--R

3~ C 00 ~
in the oroups of the formula (III) P~ substituents independently represent hydrogen, a Cl_5 straight or branched chain alkyl group, a C2_5 straight or branched chain alkenyl group, phenyl or phenyl-Cl_5alkyl group, the A ~684-5314/MR

two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), Cl_5alkyl, C1_5alk-oxy, cyano or nitro group(s), with the proviso that at least one of Ql and Q2 is other than hydrogen;
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is 0 or 1;
M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of N, M or Me, respectively;
s and r are, independently from each other, 0, 1, 2, 3 or 4, with the proviso that (i) r, s and q cannot simultaneously be 0; and (ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2, as well as pharmaceutical compositions containing these com-pounds.
From the compounds of the formula (I), those containing hydrogen as Ql and Q2 are used mainly as intermediates. ~Jhen administered into the organism, the compounds of the formula (I), independently whether they are new or not, are capable to form a stable complex with radioactive metal ions, above all with radio-active strontium and cerium being present in the blood circula-tion and extracellular space, and then to be eliminated.
From the compounds of the formula (I), the novel ones con-tain a group of the formula (III), wherein either R is other than hydrogen; or, q is 1 when R is hydrogen~ i.e. the complexes;
further, wherein M and N are other than sodium or lithium ion when q is 0.
Thus, on the one hand, the present invention relates to com-pounds of the formula (I), wherein 2~8627 nl and Q2 mean hydrogen or a group of the formula (III), with the proviso that at least one of them is other than hydrooen;
and in the oroups of the formula (III) the R substi~uents independently mean hydrogen, a Cl_5 straight or branched chain alkyl group, a C2_s straioht or branched chain alkenyl group, phenyl or phenyl-Cl_5alkyl group, the two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), Cl_5alkyl, Cl_5alk-oxy, cyano or nitro group(s);
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is O or l;
M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of N, M or Me, respectively;
s and r are, independently from each other, O, 1, 2, 3 or 4, with 2~ the proviso that (i) r, s and q cannot simultaneously be O;
(ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2;
(iii) q is 1 when R means hydrogen; and (iv) M and N are other than sodium or lithium ions when q is û.
Returning to the substituents of the formula (I), R as a Cl_5alkyl group may be of straight or branched chain such as a methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-bu-tyl, n-pentyl or isopentyl group3 preferably a methyl or ethyl group; R as a C2_5alkenyl group may be e~o. a vinyl or propenyl group; R as phenyl-Cl_5alkyl oroup may contain one of the alkyl moieties defined above, preferably a methyl group.
Me as an alkaline metal ion means preferably sodium or 2~8~27 potassium ion; Me as an alkaline earth metal ion preferably stands for calcium or magnesium ion; and Me as a transition metal ion may be e.g. the ion of a metal belonging to the 3d, 4d or 5d group, preferably iron(II) (ferrous) or zinc ions. M and N as al-kaline metal or alkaline earth metal ions are preferably the above ions; whereas the optionally substituted ammonium ion con-tains 1,2,3 or 4 above-identified alkyl, phenyl or phenylalkyl group(s) (supposed that no steric hindrance occurs). Due to their toxicity, the compounds containing a tetramethylarnmonium ion, cannot be used for administering into a living organism.
It is known that, on nuclear explosions or nuclear reactor disasters, very dangerous radioactive isotopes such as iodine-131 (131I), strontium-89 and -90 (B95r and 90Sr) as well as cesium-134 and -137 (134Cs and 137Cs~ and cerium-141 and -144 (141Ce and 144Ce) may get i nto the atmosphere /see e.g. in: Nuclear and Radiochemistry", John li~liley and Sons, pages 158 to 166 (1981/.
l~hen these isotopes get to the lungs by inhalation or to the digesting tract by intake of contaminated food or fluid or to the blood circulation or lymphatic system after resorption through the skin, they are deposited and accumulated in the tissues and finally, they lead to severe health injuries /"Summary Report on the Post Accident Review Meeting on the Chernobyl Accident"
Safety Series No. 75, IAEA, Vienna (1986)/. -After the radioactive contamination, strontium begins to be built in to the bones in several hours, and there is no more pos-sibility to elinimate (decorporate) the strontium deposited from the organism. Thus, the protection against radioactive strontium is particularly problematic.
The only possible way of protection is to inhibit the ~C fixation of strontium to the tissues, above all to bone tissues by introducing a suitable strontium-specific complexing agent to the organism, thus bindin~ in a stable form the isotope occuring in the blood circulation or extracellular space and decorporating it from the organism.

2~48627 The solving of this problem is made more difficult thereby that the calcium comp~ exes of complexing agents known from the literature, e.g. ethylenediaminetetraacetic acid or diethylene-triaminepentaacetic acid, are substantially more stable than their strontium complexes /A. Catsch: "Radioactive Metal Mobiliz-ation in Medicine", Ed. Charles C. Thomas, Springfield, Illinois (1964); A. Catsch: "Dekorporierung radioaktiver und stabiler Me-tallionen", Therapeutische Grundlagen, Ed. Thiemig, Munich (1968); A. Catsch: "Removal of Transuranium Element by Chelating ~0 Aoents", in: Diagnosis and Treatment of Incorporated Radio-nuclides, IAEA Publication No. STI/PUB/411, IAEA, \~ienna, page 295 (1976)/.
A new possibility for the research has been recognized by the synthesis of the crown ether and cryptand type molecules.
Namely, the mechanism of complex formation is in this case dif-ferent from that of the earlier known complexing agents as due to the structure of the new complex forming molecule, the metal ions get to holes with well-defined sizes and therefore, the stability of the complex formed essentially depends on the size of the metal ion.
The first promisinp results were obtained during the inves-tigations on 4,7,13,16,21,24-hexaoxa-1,10-diazabicyclo/8,B,B/-hexacosane forming a strontium complex with a stability constant of several orders of magnitude higher than that of its calcium complex /"Coordination Chemistry of Macrocyclic Compounds'1, Ed.
G.A. Melson, Plenum Press (1979)/. However, by using this com-pound in animal experiments it could only be proven that the complex formed with the ligand outside the organism was not dissociated in the organism after administration, but no evidence was obtained that radioactive strontium could be removed from the organism as a stable complex formed with the ligand. In addition, the ligand proved to be highly toxic /W.H. Muller: Naturwiss. 57, 243 (1970); W.H. Muller and U.A. MCiller: Naturwiss. 61, 455 (1974); W.H. Muller et al.: Naturwiss. 64, 96 (1977); ~. Knajfl - 6 - 2~862~

et al.: 12th Ann. Meeting of ESRB, Budapest (1976); J. Batsch et al.- Nukleonika 23, 305 (1978)/.
The compounds of the formula (I), being salts when q is 0 and complexes when q is 1, possess specific complexing properties enabling them to bind and decorporate metal ions being harmful to the organism, mainly the radioactive strontium and cerium getting to the living organism and being present in the blood circulation as well as in the extracellular space of the organism. By admin-istering the pharmaceutical compositions containing the compounds IQ of the formula (I) as active ingredients to humans or animals, the depositions of the radioactive strontium into tissue parts can be prevented and thereby severe health injuries induced by the radiation load of the or~oanism can be avoided or diminished.
F. de Jong et al. published a method for the preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16-Gdiazacycloocta-decane tetralithium salt /Rec. Trav. Chim. Pays-Bas 102, 164 (19B3)/. AccordinQ to this method, the corresponding cryptand was reacted with methyl 2-bromomalonate and the ester derivative obtained was hydrolyzed to give the lithium salt in a yield of not more than 15%. According to the British patent specification No. 2,024,822 the lithium salt thus prepared can be used in the form of a composition being useful for enhancing the solubility of barium sulfate in the petroleum industry. The corresponding tetrasodium salt is also mentioned in the same specification although it is not described in a specific example. The double salt of the tetrasodium salt with sodium bromide as well as the therapeutic utility of this double salt have been published in the Hunsarian patent application No. 2614/89.
The water-soluble salts and complexes of formula (I) accord-ing to the invention, wherein Ql, Q2, R, Me, M, N, m, n, p, r, s and q are as defined above, can be prepared on the analogy of the reaction mentioned above by reacting the corresponding halogen-ated oicarboxylic acid of the formula (II), COOH
X-C R
COOH

_ 7 _ 20 ~8 ~ 27 wherein R is as defined above and X stands for halogen, prefer-ably bromine, or a reactive derivative, suitably an ester, there-of with 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane in an organic solvent medium and then hydrolyzing the thus-obtained product by a strong base, e.g. sodium hydroxide or by a mixture of suitable molar ratio of a strong base and a hydroxide or salt, preferably a halide, of a complex-forming metal.
Alternatively, the water-soluble ~alts and complexes of the compounds of the formula (I), wherein M and N stand for hydrogen or an alkaline metal or alkaline earth metal ion and Me means an alkaline earth metal ion, can be synthetized also by reacting 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane with a 2-halodicarb-oxylic acid of formula (II) preferably with 2-bromomalonic acid in an aqueous medium at a pH of 6 to 13 in the presence of the alkaline metal or alkaline earth metal hydroxide corresponding to the salt to be prepared.
The complexes of formula (I), wherein q is 1, can be obtain-ed by reacting an alkaline metal salt of the formula (I), wherein q is 0 and M as well as N stand for alkaline Metal ions, prefer-ably the tetrasGdium salt, with an equivalent amount of a complex-forming metal halide, suitably metal chloride.
The action of the compounds of formula ~I) according to the invention, ~hich manifests itself as an increase in the elimina-tion of metal ions damaging the living organism, was investigated by using radioactive strontium or cerium ions on Swiss mice and Wistar rats of both sexes.
The elimination was studied on radioactive isotopes intro-duced in various rooutes to various sites, e.g. to the blood cir-culation, abdominal cavity, lungs, muscles or subcutaneous con-3û nective tissue of the experimental animals. The compound increas-in_ the elimination of the isotopes was administered daily once or two times in the form of an injection, powder or liquid aerosol or plaster to the organism of the experimental animals.
Thereafter, whole body activity measurements were performed and a 20~8627 retention curves were taken up ~Jhich then were compared to the results obtained in the control group.
The retention curves obtained were analysed on a computer.
"Nonlinear Regression by the Code of BMDP-3R" (BMDP Statistical Software Manual, UCLA, Los Angeles, 1~9û, Chief Ed. ll. J. Dixon) computer software was used for this purpose. It could be stated from the results obtained that the curves could be described by a two-component descending exponential function. Two data were cal-culated for characterizing the effectivity. One of these ~as the 1n so-called F factor indicatino the increase in the elimination in relation to the control and meaning the multiplication of the isotope elimination under effect of the test compound in relation to the untreated animals /see column (C) in Table 1/.
For the illustrative comparison of the compounds 3ccording to the invention additional characteristic data, the so-called EI
values are given which were obtained by multiplying the extent of isotope elimination as percentage related to the control group (effectivity, E) with the acute toxicity value (LD5U/3U, inno-cuous, I) /see column (D) in Table 1/. Although the EI value is 2û numerically not identical to the therapeutic ratio (safety lndex), it indicates in all cases to a weak or excellent activity of a product.
It is considered to be an important experimen-tal observation that no radioactive strontium if any could be de~ected in the bones of the animals treated with an active compound of the invention; a residual activity of 5 to lû% measured as whole-body retention ~Jas found in the soft parts and liver of the animals whereas a major part (65 to 7û%) of the retention measured in the organism of the control animals was built in to the bones.
3û Similar results were obtained by administering the test com-pounds in various routes to the organism.
It has been proven that an excellent effectivity was shown by compounds of the formula (I), wherein ûl and Q2 are the same and R stands for hydrogen. From these compounds N,N'-bis(dicarb-2048~27 oxymethyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane calcium complex disodium salt showed a particularly outstanding effect.
The importance of the effective compounds of formula (I) is further pronounced by their very advantageous therapeutic ratio.
Due to its low toxicity and high decorporating effectivlty, the calcium complex disodiun salt mentioned above is particularly preferred.
The compounds of formula (I) can be converted to pharmaceu-tical compositions by using the comr:on carriers and other auxili-ary materials in a known manner. The useful carriers, excipients, disintegrating, binding and other additive materials are de-scribed in detail in a number of relating handbooks.
The investigations on the effectivity of the compounds of Examples 1 to 15 showed that, after administration5 the active ingredient was absorbed and exerted a decorporating effect either in the form of an injectable solution or in the form of a sub-lingual tablet, dragée, capsule, entero-solvent tablet, powder or liquid aerosol or transdermal plaster. The effective dose was found to be 1.0 to 200 /umol/kg of body-weight, preferably 10 to 100 /umol/kg of body-weight, which was administered in one or more portions, preferably in two subdoses.
The pharmaceutical compositions containing the compounds of formula (I) as active ingredient are useful also for the preven-tion of building-in to the organism of metal ions being harmful to the living orQanism.
The invention is illustrated in detail by the aid of the following non-limiting Examples.
Example 1 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16-3û -diazacyclooctadecane tetrasodium salt a) 2.74 (14.98 mmol) of 2-bromomalonic acid are neutral-ized in 1.0 ml of water by adding sodium hydroxide solution of 7.410 moie/litre concentration in Lhe presence of phenolphthalein indicator. Then 1.75 9 (3.69 mmol) of N-dicarboxymethyl-- lo 2 0 ~8 6 2 7 -1,4,10,13-tetraoxa-7,16-diazacyclooctadecane disodium salt con-taining 14.10% by weight of sodium bromide (intermediate) obtained in a preceding reaction, then 1.95 9 (7.43 mmol) of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane are added to the above solution. The reaction mixture is maintained at 60C for 10 to 11 hours while adding 2,02 ml of sodium hydroxide solution of 7.410 mole/litre concentration (14.98 mmol) in 0.1 ml portions.
After termination of the reaction the solution is filtered off when necessary, sodium bromide is added, then it is evaporated, dried under reduced pressure and extracted in several portions with a total of 20 to 25 ml of methylene chloride.
The extract is evaporated to dryness, 10 to 15 ml of petro-leum ether are added and after filtration the precipitate is dried in a stream of nitrogen. 2.27 9 (4.82 mmol) of the product obtained containing 13.3% by weight of sodium bromide (inter-mediate) are used in the next manufacturing production batch.
The identifying data of the intermediate are as follows:
lH-NMR spectrum (200 MHz, D20, ~ ppm): 3.87 (lH, s); 3.67 (18H, m); 2.7B-2.92 (8H, m).
The residue of the methylene chloride extract is extracted with 60 ml of anhydrous ethanol until the extract is practically free from solid material. The residue of the extraction is dissolved in 6 to 7 ml of water and after adding-sodium bromide it is evaporated to dryness and dried. Thereafter, it is extracted by using 30 ml of anhydrous ethanol as described above.
The two ethanolic extracts are combined and evaporated to dryness to give 4.89 9 of double salt containing 2.71 moles of sodium bromide. The yield is 94.1% calculated for the macrocycle used.
The identifying data for the double salt are as follows:
3û IR spectrum (KBr, cm~l): 2950, 2868 (m, ~ C-H); 1605 (vs, COO/as); 1430 (m~ ~ COO/s) Other characteristic but unidentified frequencies: 1350 (s), 1320 (s), 10~5 (s), 928 (w).
- lH-NMR spectrum (200 MHz, D20, ~ppm): 4.00 (2H, s); 3.70 (8H, - 11 2 0 ~ 8 ~ 2 ~

s); 3.63 (~H, t); 2.92 (BH), t).
b) Sodium bromide is removed from the double salt by extrac-tion with 5û ml of 95% by weight ethanol. The extraction residue is dried and made free from ethanol under reduced pressure to give 3.22 9 of product. The yield is 93.2% calculated for the macrocycle used.
The identifying data of the product are as follows:
lH-NMR spectrum (2ûû MHz, D2û, ~ ppm): 3.95 (2H, s); 3.64 (8H, s); 3.60 (8H, t); 2.85 (8H, t).
13C-NMR spectrum (5û MHz, D2û, ~ppm): 179.95 (C=û); 76.45 (N-CH-(C00)2); 71.66 and 70.84 (0-CH2); 54.06 (N-CH2).
Example 2 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa--7,16-diazacyclooctadecane tetrasodium salt containing disodium hydroxy malonate The process described in Example 1 is followed by using 2.~4 9 (15.54 mmol) of 2-bromomalonic acid, 2,02 9 (4.11 mmol) of N-dicarboxymethyl-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane di-sodium salt containing 16.87% by weight of sodium bromide and 2.01 9 (7.66 mmol) of 1,4,10,13-tetraoxa-7,16-diazacycloocta-decane, except that the reaction mixture is maintained at 50C
and the sodium hydroxide is portionwise added during 10 hours.
The amount of monosubstituted intermediate obtained by extraction with methylene chloride is 2.17 9 (4.41 mmol) and contains sodium bromide in an amount of 16.93% by weight. The product weighes 3.92 ~, 'he yield is 96.5% calculated for the macrocycle used.
The product contains 1.6% by weight of disodium hydroxymalonate.
The lH-NMR spectrum of the product (200 MHz, D20, d~ppm) agrees with that of the product of Example 1, except that a resonance signal also appears at 4.31 (s) which is characteristic of hydroxymalonate.
Example 3 Preparation of N,N'-bis(dicarboxymethyl)-1,4,1û,13-tetraoxa-7,16--diazacyclooctadecane tetrapotassium salt 20~6~7 a) After neutralizing 1.32 9 (7.22 mmol) of 2-bromomalonic acid in 1.0 ml of water by adding potassium hydroxide solution of 5.760 mole/litre concentration in the presence of phenolphthalein indicator, 1.25 9 (4.77 mmol) of 1,4,10,13-tetraoxa-7,16-diaza-cyclooctadecane are added. The reaction mixture is heated at 50C
for 26 hours while adding an equivalent amount of potassium hydr-oxide solution of 5.760 mole/litre concentration in portions.
After evaporating the reaction mixture the solid residue is dried under reduced pressure and then extracted with a total of 20 ml of methylene chloride in several portions. After evaporation the residue is dried to give 0.85 9 (1.73 mmol) of N-dicarboxymethyl--1,4,10,13-tetraoxa-7,16-diazacyclooctadecane dipotassium salt containing 16.7% by weight of potassium bromide (intermediate).
This product can be used in a next manufacturing batch.
The residue of the methylene chloride extraction is extracted with 60 ml of anhydrous ethanol and after evaporation of the extract the residue is dried to give 2.18 9 of product, i.e. a yield of 94.0% calculated for the macrocycle used.
The product is a double salt formed with potassium bromide which contains 29.97% by weight of potassium bromide.
H-NMR spectrum of the intermediate (200 MHz, D20, S~ppm) 3.86 (lH, s); 3.63 (16H, m); 2.89 (4H, t); 2.78 (4H, m).
H-NMR spectrum of the double salt formed with potassium bromide (200 MHz, D20, o ppm): 3.99 (2H, s); 3.69 (8H, s); 3.63 (8H, t)S
2.86 (8H, t).
b) A pure potassium bromide-free product can be prepared by extracting the product obtained as described above with 97% by volume ethanol to give 1.16 9 of product, i.e. a yield of 75.6%
calculated for the macrocycle used.
3n lH-NMR spectrum of the title product (200 MHz, D20, ~ppm): 4.00 (2H, broad s); 3.70 (8H, broad s); 3.65 (8H, broad); 2.88 (8H, broad).

- 13 - 2 0 ~ ~ 27 Example 4 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane magnesium complex disodium salt 0.30 9 (1.46 mmol) magnesium chloride hexahydrate dissolved in 2.0 ml of water is added to a solution containing O.B1 9 (1.46 mmol) of product prepared according to Example 1 b) in 3.0 ml of water. After 30 minutes the solution is evaporated under reduced pressure and the residue is dried to give 0.93 9 (97.8%) of title product containing 17.99% by weight of sodium chloride.
lH-NMR spectrum of the title product (200 MHz, D20 in the pres-ence of NaOD, o~ ppm): 4.00 (2H, s); 3.67 (8H, s); 3.62 (8H, broad); 2.33 (8H, broad).
13C-NMR spectrum (50 MHz, D20, ~ ppm): 179.71 (C=O); 71.8a and 71.05 (Q-CH2); 54.51 (N-CH2).
t5 Note: due to deuteration the resonance signal of CH(C00)2 is absent.
Example 5 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane calcium complex disodium salt containing sodium chloride The process described in Example 4 is followed by using 0,92 g (1.65 mmol) of product obtained according to Example 1 b) and 0.24 9 (1.65 mmol) of calcium chloride dihydrate to give 1.08 y (98.2%) of title product containing 17.57% by weight of sodium chloride.
lH-NMR spectrum of the product (20D MHz, D20, ~ppm): 3.90 (4H, broad); 3.53 (14H, broad); 2.92 (4H, coalesced t); 2.72 (4H, coalesced t).
13C-NMR spectrum (50 MHz, D20, ~~ppm): 179.23 (C=O); 82.38 (_H(C00)2); 71.62 (0-CH2); 55.63 (N-CH2).
Example 6 Preparation of N,N'-bis(dicarboxy~ethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane calcium complex disodium salt containing disodium hydroxymalonate and sodium chloride - 14 - 2 ~ ~8 6 2 ~

After neutralizing 3.44 9 (18.825 mmol) of 2-bromomalonic acid in 1.0 ml of water by adding sodium hydroxide solution of 8.360 mole/litre concentration in the presence of phenolphthalein indicator, 2.010 9 (7.65 mmol) of 1,4,10,13-tetraoxa-7,16-diaza-cyclooctadecane are added. The reaction mixture is heated at 30 to 45C for 43 to 4E hours while adding sodium hydroxide solution (1.83 ml) required to form the disubstituted compound. There-after, the reaction mixture is maintained at 60C for 22 to 25 hours while portionwise adding sodium hydroxide solution in an ln amount required to hydrolyze the unreacted 2-bromomalonate. After termination of the reaction the solution is evaporated, then Example 1 b) is followed to obtain 4.720 9 of dry crude product containing 12.0% by weight of disodium malonate and practically no sodium bromide. The crude product may be worked up in any of the following two ways:
a) The crude product is dissolved in a mixture of 3.0 ml of water and 7.5 ml of calcium chloride solution of 1.000 mole/litre concentration. 12.0 ml of 99.7% by volume ethanol and O.B5 ml of calcium chloride are added to the above solution under stirring, then the ethanol content of the solution is adjusted to 90% by volume by adding 105 ml of 99.7% by volume ethanol. After vigorously stirring the suspension obtained for 30 to 60 minutes with heatino and then filtering off the solid precipitate, the filtrate is evaporated under reduced pressure and the half-dry product is dried at 75 to 85C under reduced pressure to give 4.48 9 (90.6%) of the title product containing 14.0% by weight of sodium chloride and 1.08% by weight of disodium hydroxymalonate.
The lH-NMR spectrum of the product (200 MHz, D20, ~~ppm) is in agreement with that of the product of Example 5, except that a resonance signal also appears at 4.31 ppm (lH, s) which is characteristic of hydroxymalonate.
b) The crude product is worked up as described under a) above, except that after adding the first portion (7.50 ml) of the calcium chloride solution an additional portion of 3.10 ml of 2~48627 calcium chloride solution of 1.000 mole/litre concentration is added to the reaction mixture, the ethanol content of the mixture is adjusted to 50% by volume by adding 114 ml of ethanol and the product is dried in nitrogen stream to give 5.00 9 (94.9%) of product containing also lS.5 mol% of calcium salt of the calcium complex (as calculated for the total of macrocycle) ln addition to the disodium salt of the calcium complex.
The product contains 13.2% by weight of sodium chloride, 7.35% by weight of water and a negligible amount of disodium hydroxymalonate.
The lH-NMR spectrum of the product (200 MHz, D20, ~ ppm) is in agreement with that of the product of Example 5.
Example 7 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane calcium complex diammonium 0.5 ml of water, 2.04 ml of 0.99a mole/litre calcium chloride solution and then 30 ml of anhydrous ethanol are added to 0.554 9 (l.OOQ mmol) of tetrasodium salt obtained according to Example 1 b). The solution is evaporated to about the third of an its original volume and anhydrous ethanol is added in a sufficient amount to adjust the ethanol concentration of the solution to 95-96% by volume. Thereafter the solution is warmed, stirred for 30 minutes, the sodium chloride precipitated is filtered off and washed with anhydrous ethanol. To the combined filtrate 0.554 9 (1.000 mmol) of tetrasodium salt and 0.214 y (4.000 mmol) of ammonium chloride and then water is added in an amount dissolving the solid materials. After evaporating the solution, the residue is made free from water at 75 to BOC under reduced pressure to give 1.452 y (5~.1%) of title product 3û containing 19.91% by weiyht of sodium chloride and 7.33% byweight of water. A pure chloride-free product can be prepared by extraction ~ith anhydrous ethanol.
H-NMR spectra of the chloride-free and sodium chloride-contain-ing product (200 MHz, D20, d~ppm) are identical: 3.90 (4H, broad 2o~8627 m); 3.73 (14H, broad m); 2.92 (4H, broad t); 2.72 (4H, broad t).
Example 8 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane calcium complex calcium salt After neutralizing 1.743 9 (9.53 mmol) of 2-bromomalonic acid in 2.0 m] of water by adding calcium hydroxide in portions in the presence of phenolphthalein indicator, 1.000 9 (3.Bl mmol) of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane is added. The reaction mixture is heated at 40, 45 and finally at 50C for a total of 72 hours, then the reaction mixture is heated at 60C
for 24 hours while portionswise adding o.a5 9 (11.47 mmol) of calcium hydroxide under vigorous stirring. Then, the precipitate (the major part of which is calcium hydroxymalonate) is filtered off and washed with 4 to 5 ml of water in 3 portions. The combined filtrate is evaporated under reduced pressure and then 2x35 ml of methylene chloride are distilled off from the evaporatlon residue to ob+ain a solid product which is dried at 75 to 85 under reduced pressure. In this way the title product is obtained in a yield of 2.710 9 (89.7%) with a calcium bromide content of 31.5% by weight.
The lH-NMR spectrum of this product (200 MHz, D20,S ppm) is lden-tical with that of the product prepared according to Example 5.
IR spectrum (KBr, cm~l): 2920, 2880 (m3 ~ C-H); 1615 (vs, ~ COO/as); 1450 (m, ~ COO/s) Other characteristic but unidentified frequencies: 1355, 1290 (m), 1250 (m), 10~5 (vs), 950 (m).
Example 9 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane iron(II) complex disodium salt The process described in Example 4 is followed by using 0.7~7 Q (1.35 mmol) of product prepared according to Example 1 b) and 0.26a 9 (1.35 mmol) of ferrous chloride tetrahydrate, except that the oxidation of ferrous ions to ferric ions is prevented during the preparation is prevented by uslng nitroQen atmosphere.

- 17 - 2~ ~8 6 2 7 In this way the title product is obtained in a yield of 0.835 9 (1.0%) with a sodium chloride content of 17.16% by weight.
The lH-NMR spectrum of the product cannot be evaluated due to the presence of the paramagnetic ferrous ion.
IR spectrum (Ker, cm~1): 2910, 2880 (m,~ C-H); 1630 (vs, ~COO/as); 1400 (5, ~ COO/s) Other characteristic but unidentified frequencies: 1355 (m), 1330 (m), 1100 (s), 930 (m).
Example 10 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacyclooctadecane zinc complex disodium salt The process described in Example 4 is followed by using 0.735 9 (1.32 mmol) of product prepared according to Example 1 b) and 0.18û 9 (1.32 mmol) of anhydrous zinc chloride to give 0.89 9 (97.5%) of title product containing 16.S2% by weight of sodium chloride.
lH-NMR spectrum (200 MHz, D20, o ppm): 3.6 -4.2 (18H, broad t band system); 3.1û (8H, broad t).
Example 11 Preparation of N,N'-bis(dicarboxymethyl)-1,4,10s13-tetraoxa-7,16--diazacyclooctadecane trisodium salt 1.72 ml of hydrochloric acid solution of 1.048 mole/litre concentration are added to a solution of 1.000 y ~1.804 mmol) of tetrasodium salt obtained according to Example 1 b) in 5.0 ml of water under cooling (at O to 5C), then the solution is evapor-ated, the residue is made free from water by using methylene chloride then is dried at 60C under reduced pressure to give 1.069 9 (96.7%) of title product containing 9.5% by weight of sodium chloride.
3û lH-NMR spectrum (200 MHz, D20, ~ppm): 4.18 (2H, s); 3.79 (16H, m, broad); 3.29 (8H, broad s).
The spectrum of the product taken uP in D20 in the presence of NaOD is in agreement with the spectrum given in Example 1 b).
IR spectrum (Ker, cm~1): 2940, 2850, (m, ~ C-H); 1655, 1605 (vs, - 18 - 2~862~

~COO/as); 1440 (m, ~ COO/s) Other characteristic but unidentified frequencies: 1345 (s), 1320 (s), 1120 (s), llon (5); ~30 (m).
Example 12 Preparation of N-dlcarboxymethyl-N'-(l,l'-dicarboxyethyl)-1,4,-10,13-tetraoxa-7,16-diazacyclooctadecane tetrasodium salt After neutralizing 3.003 9 (15.25 mmol) of bromomethyl-malonic acid in 0.5 ml of water by adding sodium hydroxide solu-tion of 8.360 mole/litre concentration at O to 5C in the pres-ence of phenolphthalein indicator, 2.000 9 (7.62 mmol) of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane are added. The solu-tion is maintained at 20 to 25C for 8 to 10 days while portion-wise addin~o 1.82 ml (15.25 mmol) of sodium hydroxide solution of 8.360 mole/litre concentration. After termination of the reac-tion, the mixture is stirred at 55 to 60C for 30 minutes, ~hen evaporated. The dry residue is extracted in several portions with a total volume of 25 to 30 ml of methylene dichloride. After evaporating the extract, the residue is treated with ether and the solid precipitate is filtered off. The material remaining after the extraction with ether contains also a little amount cf 1,4,10,13--tetraoxa-7,16-diazacyclooctadecane which can be removed by dissolving the product in methylene dichloride and precipitating by ether. In this way a purified N-(l,l'-dicarboxyethyl)-1,4,10,13 - tetraoxa-7,16-diazacyclooctadecane disodium salt containing 13.81% by weight of sodium bromide (intermediate) is obtained in a yield of 1.390 9 (37.2%).
After evaporation of the ethereal extract 1.159 9 of pure 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane are recovered. Thus, the actual yield of the reaction corresponds to 3B.5%.
~H-NMR spectrum of the intermediate (200 MHz, 2~ ~ ppm): 3.66 (16H, m); 2.~2 (4H, t); 2.72 (4H, t); 1.36 (3H, s).
By using the above intermediate the disubstituted derivative is obtained as follows.
After neutralizing 0.483 9 (2.64 mmol) of 2-bromomalonic 2~48~2~

acid in 0.5 ml of ~Jater by addin8 sodium hydroxide solution of 8.360 mole/litre concentration in the presence of phenolphthalein indicator, 1.002 9 (2.04 mmol) of the above intermediate contain-ing 13.81% by weight of sodium bromide are added, then the reac-tion mixture is maintained at 30 to 45C for 72 hours and at 50 to 6Q C for 24 hours while portionwise adding 0.32 ml (2.64 mmol) of sodium hydroxyde solution of 8.360 mole/litre concentra-tion. After termination of the reaction the solution is evaporated and the residue is dried at 75 to 80C under reduced pressure. After extracting the dry residue with anhydrous ethanol, the ethanolic extract is evaporated to dryness under reduced pressure and dried to give 1.323 9 (a3.D%) of the double salt formed with 2.06 moles of sodium bromideO
A sodium bromide-free product can be obtained from the double salt by extraction with 96% by volume of ethanol as described in Example 1 b) to give 0.532 9 (54.5%) of the title product.
The lH-NMR and IR spectra of the double salt are practically identical ~ith those of the title product.
IR spectrum (KBr, cm~l): 2960, 2870 (m, ~ C-H); 1645, 1600 (vs, COO/as)i 1405~ 1440 (m, ~ COO/s) Other characteristic but unidentified frequencies: 1355 (s), 1315 (s), 1095 (s), 930 (m).
lH-NMR spectrum (200 MHz, D20, ~ ppm): 3.89 (lH, S)i 3.68 (16H, m); 2.92 (4H, t); 2.7B (4H), t); 1.41 (3H, s).
Example 13 Preparation of N-dicarboxymethyl-N'-(l,l'-dicarboxypropyl)-1,4,-10,13-tetraoxa-7,16-diazacyclooctadecane tetrasodium salt sodium bromide double salt The process described in Example 12 is followed by using 1.608 9 (7.62 mmol) of 2-bromoethylmalonic acid and 1.000 9 (3.81 mmol) of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane to give 0.517 9 (26.2%) of N-(l,l'-dicarboxypropyl)-1,4,10,13-tetraoxa-- 20 - 2~8~2 1 -7,1S-diazacyclooctadecane disodlum salt containing 15.61% by weight of sodium bromide.
After evaporat on of the ethereal extract 0.50B g of 1,4,10,13-tetraoxa-7,16-diazacyclooctadecane are recovered. Thus the actual yield is 53.3%.
H-NMR spectrum of the intermediate (200 MHz, D20, d~ppm) 3.6e (16H, m); 2.B8 (4H, t); 2.82 (4H, t); 1.84 (2H, q); 0.90 (3H, t).
The title double salt containing 3.10 moles of sodium bromide is obtained in a yield of û.475 9 (52.7%) by using 0.517 9 (1.00 mol) of the above intermediate containing 15.61% by weight of sodium bro~ide and 0.229 9 (1.2Q mmol) of 2-bromo-malonic acid.
lH-NMR spectrum of the title double salt (200 MHz, D20, d~ppm) 3.8~ (lH, s); 3.67 (16H, m), 2.91 (4H, t); 2.86 (4H, t); 1.84 (2H, q); 0.88 (3H, t).
Example 14 Preparation of N-dicarboxyme~hyl-N'-(ben7yl-dicarboxymethyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane tetrasodium salt containing disodium hydroxymalonate The process described in Example 12 is followed, except that the reaction is carried out in a water/ethanol mixture of 1:1 volume ratio by using 2.081 ~ (7.62 mmol) of 2-bromo-2-benzylma-lonic acid and l.D00 9 (3.81 mmol) of 1,4,10,13-tetraoxa-7,16-di-azacyclooctadecane to obtain 0.372 9 (17.4%) of N-(benzyl-dicarb-oxy~ethyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane disodium salt con~aining 11.21% by weight of sodium bromide (inter-mediate).
After evaporation of the ethereal extract 0.714 9 of 1,4,10,13-'etraoxa-7,16-diazacyclooctadecane are recovered. Thus, the actual yield becomes 60.9%.
H-N~R spectrum of the intermediate (2ûO ~Hz, D20, d~ppm) 7.45 (2H, d); 7.29 (3H, m); 3.6B (12H, m); 3.57 (4H, t); 3.47 (2H, s);
2.78 (8H, m).
The title double salt containing 0.42 mole of disodium hydr-oxymalonate is obtained by using 0.372 9 (0.662 mmol) of the above intermediate containing 11.21% by weight of sodium bromide and 0.161 9 (û.880 mmol) of 2-bromomalonic acid. For obtaining the product, first an extraction with methylene chloride and then extraction with abs. ethanol are carried out to give the title salt in a yield of 0.432 9 (91.4%).
H-NMR spectrum of the title product (200 M~z, D20, ~'ppm): 7.43 (2H, d); 7.29 (3H, m); 3.90 (lH, s); 3.64 (16H, broad); 3.34 (2H, s); 2.Sl (4H, t); 2.80 (4H, t).
Example 15 Preparation of N,N'-bis(l,l'-dicarboxyethyl)-1,4,10,13-tetraoxa--7,16-diazacyclooctadecane tetrasodium salt After neutralizing 3.11 9 (14.79 mmol) of bromomethylmalonic acid dissolved in 1.0 ml of water by adding sodium hydroxide solution of 8.360 mole/litre concentration at 0 to 5C in the presence of phenolphthalein indicator, 3.52 9 (6.565 mmol) of N--(l,l'-dicarboxyethyl)-1,4,10,13-tetraoxa-7,16-diazacycloocta-decane disodium salt containing 18.6% by weight of sodium bromide (intermediate) are added, then the reaction mixture is maintained ~0 at 20 to 25C for 10 to 12 days while portionwise adding 1.77 ml (14.79 mmol) of sodium hydroxide solution of 8.360 mole/litre concentration. Before termination of the reaction, the mixture is heated at 55 to 60C for 30 minutes and then evaporated. The dry residue is ex-tracted in several portions with a total of 45 to 50 ml of methylene chloride. The extract is evaporated, the residue is treated with ether, the precipitate is filtered off and dried.
In this ~ay 2.0~ 9 (4.29 mmol) of intermediate containing '4.0%
by weioht of sodium bromide are recovered which can be used in a next Manufacturing batch.
The solid material remaininQ after the extraction with methylene chloride is dried at 75 to 80C under reduced pressure.
After extraction of tne dry residue with 55 to 60 ml of anhydrous ethanol, the ethanoliG extract is evaporated to dryness under reduced pressure and dried to give 3.90 9 (22.2%) of double salt 20486~7 containing 7æ.2% by weight of sodium bromide. The yield calcu-lated for the intermediate used up in the reaction is 64.3%.
H-~MR spectrum of the double salt (200 MHz, D20, ~~ppm): 3.67 (8H, s); 3.64 (8H, t); 2.73 (8H, t); 1.36 (6H, s).
A sodium bromid-free product is obtained from 3.90 9 of double szlt containing 78.2% by weight of sodium bromide by extraction with 96% by volume ethanol as described in Example 1 b). In this way 0.489 9 (57.5%) of title product is obtained.
After evaporation and drying and a repeated extraction with anhydrous ethanol, a double salt containing 80 to 9û% by weight of sod um bromide is obtained from the filtrate containing 96% by volume ethanol. This double salt can be utilized in a next manu-facturing batch.
lH-NMR spectrum of the title product (200 MHz, D20, d~ppm) 3.70 (16H, broad m); 2.71 (8H, coalesced t); 1.34 (5H, broadened s) .
Example 16 Preparation of N,N'-bis(benzyl-dicarboxymethyl)-1,4,10,13--tetraoxa-7,16-diazacyclooctadecane tetrasodium salt sodium bro-2n mide double salt The process described in Example 15 is followed; by carrying out the reaction with 0.362 9 (1.324 mmol) of 2-bromo-2--benzylma-lonic acid and 0.372 9 (û.662 mmol) of monosubstituted interme-diate containing 11.21% by weight of sodium bromide in a water/ethanol mixture of 1:1 volume ratio, a double salt contain-ing 4.2 moles of sodium bromide is obtained in a yield of 0.085 9 (11.1%).
û.2S6 9 of intermediate containing 11.21% by weight of sodium bromide is recovered. Thus, the yield calculated for the intermediate used up in the reaction is 54.2%.
H-NMR spectrum of the title product (200 MHz, D20, ~~ppm): 7.64 (4H, d); 7.45 (6H, m); 3.65 (12H, m); 3.52 (4H, t); 3.31 (4H, s);
2.62 (BH, t).

2~48627 Example 17 The acute toxicity values of the products prepared according to Examples 1 to 16 were determined on laboratory mice and rats in the following manner.
Solutions containing the compounds in various concentrations were prepared by using physiological saline solution or glucose solution of 5% by weight concentration and the active agents were administered in various concentrations into the blood circulation of the animals by injecting them slowly during 3 to 5 minutes.
lû Groups consisting of 6 to 10 animals (Swiss mice and l~istar rats of both sexes) each were used for the various dose levels.
Thereafter, the animals were observed for 30 days. The LD50/30 value (i.e. the dose causing the death of 50% of the experimental animals within 30 days) was determined from the number of animals died during this period by using probit analysis /D.J. Finney:
Probit Analysis (2nd ed.) Cambridge University Press (1952)/.
These values expressed in mmol/kg of body-~eight for the active agents according to the invention are summarized in column (B) of Table 1.
Table 1 Characteristic data of the products according to the invention (A) (B) (C) (D) ExampleAcute toxicity F factorEI value No. LD5P/~30 mmol/K~ _ l/a 0.327 1.8 5.40 l/b 0.379 2.8 9.66 2 0.368 3.3 ll.lû
3/a 0.333 2.3 12.30 3/b 0.272 1.7 8.90 4 0.211 2.~ 7.3û
above 2.5 3.7 145.00 6/a above 3.0 3.9 158.00 6/b above 3.0 4.2 160.00 20~8~27 - 2~ -7 above 2.5 1.7 65.00 S above 1.5 1.,, 5~.00 ~ 0.?50 1.7 ~.25 'Q 0.~91 2.3 3.20 11 0.1'1 2.6 2.S0 1~ 0.305 1.5 7.'3C
13 0.302 2.1 13.30 1- 0.. 22 l.~r 7.60 0.~50 1.3 ',.E0 ln li 0.~70 1.3 7.10 '~xa~ple lS
The radioactive isotope eli~in2tion-incre3sins effects OT
compounds prepared according to Fxamples 1 to 16 were cmpared on Swiss mice as described hereinafter.
An activity OI 37 to 74 kZq (1 to 2 /uCi) of r~d7Oactive strontium (Sr-85512) or radioactive cerium (Ce-14'~C13), respectively, was administored into the abdomlnal cavity of the animals which were then divided into treatment oroups consistino of 5 to 10 ani~als each. At 30 to 60 minutes after admlnistration of the isotope, the animals OI the treated _roups ~ere intra-venously (i.v.) p~iven 'he active ao,ent used in an amount to achieve a concentration of 50 to 100 /umol/kg of body-weight in the ani~al or~anism or each treatment. The animals of the con-trol eroups were simil2rly treated with the carrier ~sterile phy-siological saline or Qlucose solution of 5% by weight concentra-ion) ~ithout active a_ent.
The a~ount of radioactivity introduced to the animal organ-ism ~.~as deter~ined im~ediately after ad~inictrat,on of the iso-.ope, then these measurenen+s were repeated daily or in every t~o or thrPe days in a devlce constructed lor the pur,oose of whole-bor'y me3sure~ents on s~all ani~als. The counts observed ~;Jere related to the startino activity value of the zero (û) day and a so-called retention/tl~e correlation ~!as obtained by consideriny ~he activity retained in the organism. The change of activity of 20~8~7 the animal organism in the time is illustrated in Figure 1 where the time (in days) elapsed after the treatment is shown on the abscissa and the whole-body retention as percentag~ is plotted on the ordinate. It can be seen that the rate of elimination of Sr-65 administered in-to the abdominal cavity of control animals was slow: within 1 day only 15%, within 4 days 25% and within 7 days 30% of the activity were eliminated from their organism and the elimination was even more delayed in the later period. On the contrary, within 1 day 40%, within 4 days 65% and within 7 days 67% of the radioactivity introduced were excreted from the animal oroanism after a single treatment with 100 /umol/kg of the active aoent prepared according to Example 13. Even more advantageous results were obtained by using the product prepared according to Example 5 which gave elimination values of 81, 84 and 85%, respectively, within the intervals mentioned above. 8ased on analysis carried out in the above manner, it was stated that the retention curves could be described by a two-component descending exponential function. The F factors and EI values relating to the products according to the Examples mentioned above are summarized 2n in the columns (C) and (D), respectively, of Table 1. It isobvious that the effectivities of various decorporating agents, particularly on the basis of their EI values, are highly differ-ent. In our opinion, the agents with an EI value-of O to 5 are weak, those with an EI value of 5 to 50 have a medium effect, those with an EI value of 5Q to 100 are good and those with an EI
value above 100 are excellent.
Example 19 a) The whole-body retention curves illustrated in Fioure 2 show the elimination o~ radioactive strontium introduced through ~O the trachea into the lungs of Wistar rats after intraperitoneal (i.p.) administration of the compound described in Example 6 b).
It can be seen from the pattern of the upper curve of Figure 2 that the radioactive strontium was eliminated to a 10PJ grade from the organism of control animals treated only with the solvent.

- 26 - 2~48~2 7 ~ot more than 30 to 35% of the starting activity were eliminated in the days following the inner contamination. On the contrary, the whole-body load significantly decreased from ~0% (measured on the controls) to 20% both in the group treated once with 5D
/umol/kg of body weight of the agent according to the invention (middle curve) or in the group treated twice in 3-hour interwal (lower curve) within 1 day following the treatment. The rapid elimination of the isotope was continued in the days following the treatment, and reached &~ to SO% in the once-treated group lG and 94 to 96% in the twice-treated group. It is also an important experimental result that hardly any or no radioactive strontium could be detected in the bones of the animals treated with the active aoent according to the invention and killed at the end of the experiment. The residual activity of 5 to 10% measured as whole-body retention was found in the soft parts and liver of the animals whereas the maJor part (65 to 70%) of the retention found in the controls were built in to the bones. Similarly advantage-ous results were obtained by administering the compounds accord-ing ~o the invention in an intravenous route or into the subcuta-neous connective tissue.
b3 The compounds of the invention increasing the isotope elimination were tried to remove other radioactive metals, firstly cerium-144 (belonging to the group of the rare earth elements) from the animal organism. In this Example the elimina-tion of Ce-144C13, introduced to the lungs of female Wistar rats, i5 illustrated as a function of the time elapsed after a sinole dose or after a treatment repeated at a 24-hour interval (see Figure 3). The results obtained prove that the compounds accord-ing to the invention can advantageously be used also for the removal of this radioactive compound having a relatively low solubility in body fluids from the lungs. At the end of the experiment, i.e. on the 30th day, 40% of the introduced starting activity were present in the control animals whereas 14% were detected in the group treated i.p. once ~ith the active agent according to the invention; and the whole-body retention decreased to 5.6% on effect of treatments with the active agent by 60 minutes and then by 24 hours after administration of the isotope. The beginning abrupt but later delayed pattern of whole-body retention curves of the treated groups is likely due to solubility relations of the radioactive contaminating compound and the elimination properties of the metal complex.
Example 20 The effect of compounds according to the invention on the elimination of isotopes after administration into the blood cir-culation, abdominal cavity or subcutaneous connective tissue have been described in Example 17 to 19. From the viewpoint of human therapy and even more of the effective and rapid protec~ion of a greater population it was important to justify that the compounds according to the invention are useful to decorporate internal radioisotope contaminations also by other routes of administra-tion.
I~istar rats were used for this purpose. In a part of the examinations radiostrontium was administered into the abdominal cavity of the animals and after 60 minutes the active agent according to the invention W25 given through the rachea into the lungs. The compounds indicated in Table 1 with an EI value above 100 were used as active agents. Thereafter, the whole-body activ-ities were measured for 30 days. It turned out on evaluation of the whole-body retention curves, plotted in the usual manner, that the active agents according to the invention exerted a good effect on the elimination after inhalation in a powder or liquid aerosol form. The retention of 91% measured in the control ani-mals was diminished to 15% on the first day, and became lower than 10% on the 3rd day in the treated animals. An F factor value of 7.9 and EI value of 164 (c.f. Table 1) were found which sup-ported the above statements.
The possibility of adsorption of the active agents according to the invention from t~e epithelial surface was studied in addi-tional experiments. A 3x3 cm area of the dorsal skin of rats ~as depilated. After administering radioactive strontium through the trachea into the lungs of the anaesthetized animals, whole-body 2ctivities were measured, then the active agent l~as applied in solution on the dorsal skin area previously depilated and cleaned. The area containing the active a~oent was covered with an adhesive plaster. It could be stated from daily measurements of ~he whole-body activites that the active agents according to the invention exterted their isotope-mobilizing effect through the skin, too. By considerino all the measurement values, an F factor v21ue of 2.5 and EI value of 105 to 110 were obtained.
On the basis of the above exper mental results the active agents according to tnhe invention can be used also in the form of pharmaceutical compositions such as sublingual tablet, supposi-tory, enterosolvent dragée or capsule or transdermal plaster.

Claims (10)

Claims:

1. Compounds of the formula (I), (I) wherein Q1 and Q2 mean hydrogen or a group of the formula (III), with the proviso that at least one of them is other than hydrogen;
and in the groups of the formula (III), the R substituents independently mean hydrogen, a C1-5 straight or branched chain alkyl group, a C2-5 straight or branched chain alkenyl group, phenyl or phenyl-C1-5alkyl group, the two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), C1-5alkyl, C1-5alk-oxy, cyano or nitro group(s);
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is 0 or 1;
M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of M, N or Me, respectively;
s and r are, independently from each other, 0, 1, 2, 3 or 4, with the proviso that (i) r, s and q cannot simultaneously be 0;
(ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2;
(iii) q is 1 when R means hydrogen; and (iv) M and N are other than sodium or lithium ions when q is 0.

2. A compound of the formula (I) as claimed in claim 1, wherein Q1 and Q2 are the same and R means hydrogen.

3.N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diaza-cyclooctadecane calcium complex disodium salt.

4. A composition useful for the decorporation of metal ions, mainly radioactive isotopes, being harmful to the living organ-ism, which comprises an effective amount of at least one compound of the formula (I), (I) wherein Q1 and Q2 mean hydrogen or a group of the formula (III), in the groups of the formula (III) R substituents independently represent hydrogen, a C1-5 straight or branched chain alkyl group, a C2-5 straight or branched chain alkenyl group, phenyl or phenyl-C1-5alkyl group, -the two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), C1-5alkyl, C1-5alk-oxy, cyano or nitro group(s), with the proviso that at least one of Q1 and Q2 is other than hydrogen;
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is 0 or 1;
M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of M, N or Me, respectively;
s and r are, independently from each other, 0, 1, 2, 3 or 4, with the proviso that (i) r, s and q cannot simultaneously be O; and (ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2.

5. A composition as claimed in claim 4 characterized by a tablet, dragée, capsule, suppository, injectable solution, powder or liquid aerosol or transdermal plaster form.

6. A composition as claimed in claim 4 or 5, which comprises as active ingredient a compound of the formula (I), wherein Q1 and Q2 are the same and R stands for hydrogen.

7. A composition as claimed in claim 4 or 5, which comprises N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16--diazacycloocta-decane calcium complex disodium salt as active ingredient.

8. Use of the compounds of the formula (I), (I) wherein Q1 and Q2 mean hydrogen or a group of the formula (III), (111) in the groops of formula (III) the R substituent independently represent hydrogen, a C1-5 straight or branched chain alkyl group, a C2-5 straight or branched chain alkenyl group, phenyl or phenyl-C1-5alkyl group, the two latter ones optionally being substituted on their aro-matic part by one or more halogen(s), C1-5alkyl, C1-5alk-oxy, cyano or nitro group(s), with the proviso that at least one of Q1 and Q2 is other than hydrogen;
Me stands for an alkaline metal or alkaline earth metal or transition metal ion;
q is 0 or 1;
M and N, independently from each other, stand for hydrogen or an alkaline metal, alkaline earth metal or optionally substi-tuted ammonium ion;
m, n and p are integers each being equal to the charge of M, N or Me, respectively;
s and r are, independently from each other, O, 1, 2, 3 or 4, with the proviso that (i) r, s and q cannot simultaneously be O; and (ii) the number of hydrogens in the meaning of M or N may be 0, 1 or 2 for the decorporation of metal ions, mainly radioactive isotopes, being harmful to the living organism.

9. Use of a compound of the formula (I), wherein Q1 and Q2 are the same and R means hydrogen, for the decorporation of metal ions, mainly radioactive isotopes, being harmful to the living organism.

10. Use of N,N'-bis(dicarboxymethyl)-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane calcium complex disodium salt for the decorporation of metal ions, mainly radioactive isotopes, being harmful to the living organism.