JPH04500218A - Method for producing active ingredients and pharmaceutical compositions for dissociating radioactive isotopes from living organisms - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Method for producing active ingredients and pharmaceutical compositions for dissociating radioactive isotopes from living organisms The present invention is for dissociating radioactive isotopes from living organisms. The present invention relates to the production of active ingredients and pharmaceutical compositions.

Nuclear fission in experimental, isotope producing or energy supply reactors and testing of nuclear weapons. The majority of these high-temperature materials undergo nuclear fission, with the production of significant amounts of radioactive byproducts. products as well as highly hazardous radioactive isotopes, such as iodine 7.131, Trontium-89-90, Cesium-134, Cesium-137, Cerium- 141 and cerium-e-44. these are in the atmosphere If released, radioactive contamination can occur in living organisms.

The route by which these isotopes can enter the human body (inhaled with the air) is respiratory system (taken in with food and drink); digestive system (taken in with food and drink); There are three types of epidermis (contact with intact skin).

Considerable methods are available to reduce or prevent damage to health. ing. However, appropriate adsorbents can be administered orally to prevent gastrointestinal absorption. Only a few isotopes, primarily radiostrontium, are present. Medical relief contaminated If started late, several hours after the Of the radioactive isotopes transported by the flow, the absorbed fτ portion is absorbed into the bones. Acts on the attachment of isotopes, preventing their systematic binding and promoting their dissociation. There is no effective method.

This suggests that radiostrontium in the blood and outside other cells can be converted into a stable complex. Research on highly effective human and dedicated drugs that can be combined in the form of Research is underway. This prevents tissue deposition of isotopes and also natural excretion (feces, urine) will become possible.

The following requirements are specified for such drugs:

(a) The production of one body is caused by the coexistence of a large number of ) in biological systems even in the presence of ions (e.g. Ca', Na', Ni', etc.) and ligands. arise.

(b) The toxicity of the drug is acceptably low (broad effectiveness).

(c) The drug is water-soluble.

(d) The drug can also be administered parenterally.

In the early 1950s, when complex chemistry began to make rapid progress, readily available organic compounds Does not form complexes with ligands and inorganic ligands, or has a very high stability constant (formation constant). low alkalinity, maintained solely by electrostatic interactions between metal ions and ligands Complexes of transition metals have been mainly studied, ignoring complexes of metals and alkaline earth metals. there was. This concept was introduced in the late 1960s by the “Crown Ether” and “Cryptate” arrangements. It was shattered by Ishi's discovery.

Crown ethers mainly contain oxygen atoms as donor atoms, but crib ethers mainly contain oxygen atoms as donor atoms. The catalyst contains oxygen and nitrogen donors. These are structurally well dimensioned. It holds the metal ions taken into the pores. This is only for metals of certain dimensions. can form stable complexes with these types of ligands. Therefore, Crown A Tel and cributate are much more specific than known ligands. These ligands The production constant (10 gK) with an aqueous alkaline earth metal solution is shown on the surface dination Chemistry of Macrocyclic Co mpouncls.

Ed, [1;, ^, Melson, Plenum Press, 197 9).

议± Ligand Ca” Sr” Ba” 15-crown-50,52,723,87 dicyclohexyl-18-crown -60,42,643,27 dibenzo 18-crown -61,001,951 ,10-diaza-4,7,13,16-chitraoxycyclooctadecane 2.56 2.97 Cribteto (2,1,1>2.50 2.00 2. 00 Cryptate - (2,2,i) 6.95 7.35 6.30 Cryptate To (2,2,2) 4.40 8.00 9.50 Cribtate (3,2,2 ) 2.00 3.40 6.00 Cryptote (3,3,2) 2.00 2.00 3.65 The data in Table ■ shows that the size of the “vacancy” in the ligand is significantly affected by the production constant It shows that it is working. In some cases, for example, Cribteto (2,2 , 2), the production constant for Sr'' is much higher than that for Ca''. There is.

Since the above data was obtained, 5r-85, Ra-224, Pb-212°B Regarding the removal of a-140 and La-140 isotopes from living organisms, )-(2,2,2)(4,7,13,16,21,24-hexaoxa-1゜1 Animal experiments were conducted using 0-diazabicyclo-/S, 8.8/-hexacosane (W, H, Mjiller, Naturwiss, 57.248/19 70/; N, 11. Miller and IA, ^, Miller, Naturwiss, 61.455/1974/; JH, Miiller et al, Naturwiss, 64.96/19777'; J.

Knajfl et if,, 12th ^nn, Meeting of ES RB, Budapest.

1976; J, Barsch, J, Eisler and Z, 5zot, Nukleonika, 305/1978/), metal complexes in vitro The concept of producing a complex, then administering it subcutaneously and investigating the excretion of the complex The value of experimental results has decreased. These experiments remove the release that is already present in the animal. The radioactive isotope undergoes an in vivo formation reaction with the subsequently administered ligand, resulting in its own formation. Rather than forming a stable complex that can be dissociated from the body by natural excretion methods, the radioactive metal and that dissociation of complexes generated externally from ligands does not occur in complex biological systems. That could only be proven. Based on practical experience, we have shown that the ligand compound is more effective than the ligand itself. It should be noted that the complex form is much less toxic.

In contrast to known complexes, 1,10-diaza-4 synthesized from monocyclic cributate ,7,13,16-titraoxacyclooctadecane~N,N'-diacetic acid rare earth The formation constant of complexes of similar metals decreases as the number of atoms increases (ion size decreases). decreased. However, the formation constants of complexes with Ca'' and Sr'' are within the experimental error range. (8.39 and 8.29 respectively) (C, ^, Chang .

Inorg, Cbev, 25.355/1986/) described acyclic aminopolymer Complexes of carboxylic acids (EDT^, ethylenediaminetetraic acid [IQ, DTP^.

(diethylenetriaminepentaacetic acid, etc.) is considerably more stable with Ca than with Sr. ing.

[j] of the present invention is preferably bonded to the [I] macrocycle to form the invivo base of the complex. In order to use 1 for the qualitative analysis, one tree of o-ring kributi-1, coordination r and soh was prepared. It was to build. The final [] target is Fujios 1 ~ Rontium, depending on the case Other radioactive gold, other isotopes and living organisms c) Remove al derivatives and obtain Delir. , from some experimental data, 1,4.10j3-te1-raokijouto16 -Diasafuchrooctatecan-N,N'-cimaronic acid, 7) Tetof)'- The active agent based on Ium Shioto is administered to animals (J) in the body, and (F) to species 2 sites (peritoneal cavity, skin). Radiosl-■contium and radiocerium administered to the lower interstitial tissues, lungs) It has been proven that it can promote excretion.

The method of the present invention comprises The novel method consists of three methods for the preparation of compositions containing rooctadecane cimalonate.

The preparation of such compounds is described by F. de Jong et al. (Re cl, Trav, Cbi+n, Pays-Bas, 102.16417 37'1983/). In this method, in order to replace hydrogen atoms with nitrogen atoms, 1, ．． 4.10.13-Tetraoxa-7,1O-diazacyclooctadecane to α- react with a halogenated methylmalonate ester and then convert the ester into a lithium salt. It is hydrolyzed into

To replace a hydrogen atom with a nitrogen atom, α-brominated dinato-11ummalo-1 is It has been found that the halogenated methyl chloride is preferable to the halogenated methyl chloride. In this case, add water No decomposition is necessary and the water-soluble salt is obtained directly. Furthermore, the sodium salt is lithium In contrast to salts, it is non-hygroscopic and therefore more convenient for the production of pharmaceutical compositions, etc. It is an ingredient. Another advantage of sodium salts over lithium salts is their lower price. be.

In the method of the present invention, 1,4,1.0.13-tetraoxy-7,16-di Reacting azacyclooctadecane with 2-bromomalonic acid disodium salt, 1.4,10.13-tetraoxa-7,16-diazacyclooctadeca>'- An active agent containing N,N'-cimalonic acid tetrasodium salt is prepared. Like Alternatively, the reaction mixture is carried out in a slightly alkaline aqueous medium at 70-80°C. Check the alkalinity appropriately with a phenolphthalein indicator and keep the mixture pale during the reaction. Adjust to a bright pink color and maintain this state.

1.4,10.13-tetraoxy-7,16-diazacyclooctadecane-N , the active agent containing N'-cimaronic acid tetrasodium salt is bioavailable. Combines radioactive metal isotopes, mainly radiostrontium and radiocerium can do. Stable complexes generated in vivo can be produced using natural methods. It can be dissociated from the body in the form of a complex. The pharmaceutical composition of the present invention 1,4,10.13-tetraoxa-7,16-dia produced based on the invention method Active ingredient containing cyclooctadecane-N,N'-cimalonic acid tetrasodium salt and a pharmaceutically acceptable carrier such as normal saline (nor+l1als). aline 5 solution) or 5% glucose solution by volume) .

The composition preferably contains 100 to 100% of the active ingredient mixed with a 5% by volume glucose solution. 500 mg, preferably 2501.

The healing power of the composition was demonstrated by animal experiments. In this way, the minimum lethal dose and normal The mean lethal doses (LD, , and LDso) were established.

To calculate the lethal dose, increasing concentrations of the active ingredient were administered intravenously to the animals.

The minimum lethal dose and average lethal dose were calculated from the mortality rate within 30 days. Based on Example 1 1,4,10.13-tetraoxa-7,16-diazacyclooctane prepared by Tadecane-N,N'-cimalonate tetrasodium (DMCRYP) salt x NaB LD per kg of body weight of active agent containing r (x=2.5-8). 73 The value was 1.05 mmole. In any case, in this experiment this dose one-tenth of the total amount was administered to each animal.

About DMCRYP's No. 1 Sokei n Kaishichi 0 sets of pharmaceutical compositions containing DMCRYP active ingredients expressed as "PTR-23" The composition is prepared in a carrier, preferably sterile normal saline or a 5% by weight glucose solution. It contained liquid. The preferred ratio of active agent per carrier l<em>3 is between 100 and 500 It was mg.

Enhancement of dissociation of radiostrontium and rare earth metal radiocerium from the body To test the effects of DMCRYP on male and female Swiss mice, Wistar rats were selected. Experiments generally involve radioactivity of 37 to 54 kBQ ( 1-2 μCi) of radiostrontium (5rCI□) or radiocerium 1"'CeCl, , lungs) and then introduces 100 μ5 ole/kg of active agent in a single injection. PTR-23 was injected intravenously 30 to 60 minutes later. in a comparative test is a known polyaminopolycarboxylic acid type dissociating agent, DTP^ (diethylenetriamine). Calcium-trisodium salt of Nopenta Vinegar R) (Heyl and Co., Berlin) was used at equimolar concentrations. Measure the radioactivity of the whole body every 1 to 4 days. Calculate the amount of residual isotope in the animal body by 1. Expressed as % of radioactivity Ta.

One dose of PTR-23 for enhanced excretion of 5r-85 isotope introduced into the peritoneal cavity Figure 1 shows the effects of intravenous administration (day 0). Figure 1 shows the pattern of residual isotopes in the animal body. - Centage (residual rate throughout the body) is plotted against the day of the experiment. same The effectiveness of treatment with compounds of the invention carried out 30 minutes after topical administration was determined by the corresponding Shown by the steep drop in the line. It reached 35.8% on day 1, but this value was lower than that of control animals. It is less than half of the 5r-85 content of 73.2%. This ratio varies throughout the experiment. after treatment with PTR-23 compared to 45.5% of the untreated control. reached 212%. This series of experiments further demonstrated that the dissociating agent DTPA was not present in the animal's body. The results show that the removal of Sr isotopes is ineffective (^, Catsch: 　DekorporierunB　radioaktiverund　5tab Iler Metal! 1onen, K., Thiemig Verlag, 　Miinchen.

196B).

The invention will be particularly illustrated by the following examples. The following examples limit the scope of the invention. It's not something you do.

Example 1 shows the preparation of the active agent. Examples 2 to 6 were prepared from this active agent. This shows the healing power of the prepared pharmaceutical composition.

Example 1 1.4,10J3-tetraoxa-7,16-diazacyclooctadecane-N, Production of active agent containing N'-cimaronic acid tetrasodium salt ([)MCRYP) Dissolve 2.80 g < 15.3 mmoie) of 2-bromomalonic acid in 2 cm’ of water. This solution was then titrated by adding a 1.5-288aOB solution. Phenolphthalene When one drop of In indicator was added, it turned pale pink. 1. OOg (3,81 mmo 1,4,10,13-tetraoxy-7,16-diazacyclooctade (Kryptofix 22. Merck) was added to the solution. reaction mixture was maintained at 75-80°C for 14 hours, while 1.8738 of 8.55ctaff The NiOH solution was added dropwise from the bilulet to maintain the pink color. It was evaporated and vacuum dehydrated on a water bath at 80° C. for 6 hours. Dichlorinate the residue with 1501 Collected with methane, blotted, extracted three times with dichloromethane, and dried with a stream of nitrogen.

Dissolve the white solid with absolute ethanol until the material is not appreciably soluble (15 to 17 times). The product was extracted. A white precipitate precipitated from the extract during the extraction. ethanol The extract was evaporated, collected with dichloromethane, filtered and extracted three times with dichloromethane. and dried with a stream of nitrogen. Product yield was 1.447g.

The dichloromethane extract was evaporated with a stream of nitrogen to give highly hygroscopic yellowish crystals. 357g was obtained.

The residue (white precipitate) from the ethanol extraction was dissolved in 20 cm' of water, The temperature was maintained at 80°C for minutes. Evaporate the solution in vacuo and dehydrate in vacuo on a water bath at 80°C for 5 hours. did. Other processing is as described above. Evaporate the ethanolic extract in vacuo and dichloromethane. Collected with methane, filtered and dried with a stream of nitrogen. A solid of 0.430B was obtained. It was.

The products from the ethanolic extracts were combined, suspended in ethanol and incubated at 70°C for 3 The mixture was stirred for 0 min. The mixture was then evaporated, collected with dichloromethane and filtered. and dried with a stream of nitrogen. Purchase of product was 1.830 g with 58% yield .

Product foil 33 weight? 6) 1,4,10゜13- Tetraoxa-7,16-diazacyclooctadecane-N,N'-cimalonic acid tetra It is a double salt of torasodium salt and sodium bromide.

■ Characteristic IR band (KBr), C/C-2950.2868 (+e, C/C- H/) 1605 (vs, ν/COO8,) 1430 ( Theory, ν　/COO,) Characteristics unconfirmed IR band: 1350(s), 1320(s), 101095(s)928( 'HNMR data (D20), pp+o: 2.92 (t, 8H, N-Cl1 2) 3.63 (t, 8H, 0-C112) 3.70 (sg, 8H, 0-CR2-CBz-0) 4.00 (sg, 2H, N- C)l).

``丞'' L1 ni1 dissolves well.

Example 2 The results in female Wistar rats are shown in Figure 2. The experimental process was as described above. It is written in the same way as in Figure 1. The only difference is the apposition into the IIIM cavity. Compositions of the invention, wherein the active agent is administered intravenously 6 days after introduction into the body. is even more effective in rats. As a result of using the new armor punishment, on the first day after processing The amount of radiostrontillam in the animal's body was more than three times that of the control, 69.6%. Compared to this, it decreased to 22.0%. Similar to the results obtained with the mouse, the ratio between the curves is There was no variation throughout the experiment, and at the end of the study, there was no variation between treated and control animals. The residual rates reached 13.5% and 36.8%, respectively. Glue treated with DTPA The residual values in the loop were lower than those in the control, but the difference was statistically significant! Tasty too It wasn't.

Once the active agent is incorporated into the bloodstream 60 minutes after isotope administration, composition PTR- 23 significantly promoted the dissociation of intraperitoneally administered 5r-85, thereby The amount of radioactive isotope in the body of an animal is reduced after receiving a single administration of a small amount of the isotope. The experimental data for Example 2 were explicit.

Example 3 In this series of experiments, PTR-23 was administered once intravenously to the skin or skin. We wanted to investigate the possibility of dissociation of radiostrontium that had entered the lower interstitial tissue. 0).

The result was radiostrontium that entered through a wound in the epidermis (puncture, bruise, cut). It was shown that the amount of Wi was significantly reduced by the PTR-23 composition. from infection When treated after 30 minutes, the residual value in the whole body of the animal at the first time was 56.9% of the control. In contrast, it was 31.5% (almost 50% better); this ratio There was no variation throughout. Treatment with DTPA had no effect.

Example 4 The whole body residual curve in FIG. of radiostrontium taken into the lungs of Wistar rats through the trachea at Regarding removal. The effectiveness of the active agent is not dependent on the route of administration, i.e. intravenous administration or intraperitoneal administration. Experimental data has conclusively proven that the same effect can be obtained by treatment within the membrane tissue. It was.

The effect of the composition of the present invention on promotion of Sr dissociation is shown in the lower two curves of FIG. The amount of Sr isotope taken up into the lungs sharply decreases after administration of the above composition. I understand. At the first time, the amount of isotope decreased to 48.2%. Isotope in contrast The amount was 88.3%. The biggest difference was observed at the 188th time, and at that point The residual values in the treated and control groups were 28.6% and 58.6%, respectively. It was 7%.

Example 5 The dose-dependent effects of the dissociating agent of the invention were determined in Swiss mice. Residual curve in Figure 5 after excretion of Sr isotope taken up into the peritoneum (10-100 μaolez’k s) at the active agent concentration. At concentrations as low as 10 μmole/kg, the body Experimental data show that the amount of isotope remaining in the sample is significantly lower than that in the control. ・It became clear in the evening. The effect increased with dose, but above 50 μmole, kg Then, the residual value at the end of the experiment (300th test) was 40.6% for the control, and then 34.3% in order of increasing dose.

28.3%, 25.2%, 23.9%, 21.8%. Enhancement of excretion is no longer proportional to the amount of active agent. Therefore, radiostro 〉Repeated administration of smaller doses is reasonable to treat internal infections of tium. It seems to be.

Example 6 Radioactive metals other than radiostrontium, mainly rare earth metals The pharmaceutical composition of the present invention was tested for removal from the body of 144 animals, and 62 and seri Um.-144 isotope was introduced into the peritoneum of female Swiss mice. 30 minutes later this Mice were treated intravenously with 100 μl of PTR-23 or DTPA. 2.

On day 28 and day 4, the one-sentence treatment was repeated. The results are shown in Figure 6. The whole body residual properties for excretion indicate that the dissociation of active isotopes from the animals is different from that in the control group. This indicates that the process is being carried out relatively slowly. 1st time by DTPA Treatment resulted in a slight decrease in residual values on the second day (72.8% vs. 85.9%). ), but the second and third treatments had no effect. That is, the following two curves The flow was the same.

On the contrary, residual radiation in the body (during the second treatment on the second day after administration of the composition of the invention) The performance value was only 56.2%, in contrast to the above 85.9%. 2nd and 3rd By performing the second treatment, the difference widened until the 7th day. PTR’-2 at day 7 The amount in the 3 groups was 33.4%, while the amount in the control was 76.3% (immediately From this point on, the excretion (dissociation) rate was the same, Therefore, the curved lines became parallel.

1.440.13-tetraoxa-7,16-diazacyclooctadecane-N, Composition containing N''-dimalo〉acid tetrasodium salt (DMCRYP)\'! 5 (PTR-23) is preferred for mobilization of radiostrontium and radiocerium. This composition has been equivocally proven by practice to work effectively. It is effective when the sexual isotope enters the peritoneal cavity, subcutaneous interstitial tissue or lungs. hundreds of animals A study of test animals revealed that there were no harmful side effects, and Therefore, the composition of the present invention is expected to be suitable for treating patients contaminated with radioactive isotopes. It will be done.

As concluded from the data in Table ■, radiostrontium from living organisms Early studies with cributate-(2,2,2) ligands for the removal of , the behavior of a metal/ligand complex produced in vitro is only observed when administered into the body. Not investigated (Mjller, 1970; , 1974 and 1977; najfl et al, 1976) , J, Batsch et al. (Nukleonika 23゜305/197B/ ) only administered radiostrontium intravenously to rats for 0.5 hours. , 2 hours, 4 hours, 24 hours, 48 hours, 72 hours, 192 hours and 216 hours Later, the experimental process of intraperitoneal treatment and testing of cribtate (2°2.2) was carried out. Reporting. 10-80 II+ per animal to achieve significant dissociation 8, a very large amount of activator is required. This is the same as semi-lethal doses (LDs.7..) etc., and the amount even exceeds that amount. Lll,. Criticism against rats regarding It was revealed that the acute toxicity of butate-(2,2,2) is 292 μmole/kg. It should be noted (1. C, R, P.

Publication No. 20. p, 76/1972/几These places Even in the case of microscopy, the reduction in residual rate never exceeded 10-12%. child In contrast, with the composition of the present invention, the radioactivity I in rats was reduced to 4 at a dose 1/10 of the semi-lethal 1. 9.6% (see Example 2 and Figure 6), the efficacy of the composition of the present invention is extremely high. This shows that it is preferable for

Days after processing FIG.4 Days after processing FIG. 2 ! To! Days after 1m Days after processing FIG.5 Days after processing 1”lG, 6 international search report

Claims (6)

[Claims] 1.1,4,10,13-tetraoxa-7,16-diazacyclooctadecane is reacted with 2-bromomalonic acid disodium salt and the resulting product is isolated. A method for producing an activator suitable for dissociating radioactive isotopes from living organisms, characterized by Law. 2. Characterized by carrying out the reaction in a slightly alkaline aqueous medium at 70-80°C The method according to claim 1. 3. Test the alkalinity of the reaction mixture by adding a phenolphthalein indicator to the system. , wherein the mixture remains pale pink during the reaction. Method described. 4. An active agent produced according to the method according to any one of claims 1 to 3. 1. A pharmaceutical composition for dissociating a radioactive metal isotope from a living body. 5. It further comprises a carrier which is sterile normal saline or a 5% by volume glucose solution. The pharmaceutical composition according to claim 4, characterized in that it comprises: 100-500 dissolved in 1 cm3 of carrier which is 6.5% by volume glucose solution. Pharmaceutical composition according to claim 4 or 5, characterized in that it contains mg of active agent. .