WO1992004056A1

WO1992004056A1 - USE OF RADIOACTIVE METAL ION COMPLEXES WITH all-cis-1,3,5-TRIAMINO-2,4,6-CYCLOHEXANTRIOL AND THEIR DERIVATES FOR X-RAY DIAGNOSIS AND IN TUMOR THERAPY AS WELL AS FOR PRODUCING X-RAY DIAGNOSTIC AND TUMOR THERAPY AGENTS

Info

Publication number: WO1992004056A1
Application number: PCT/EP1991/001679
Authority: WO
Inventors: Isidor Erni; Walter Schneider; Hans Kaspar HEGGETSCHWEILER; Peter Geisser
Original assignee: Laboratorien Hausmann Ag
Priority date: 1990-09-05
Filing date: 1991-09-05
Publication date: 1992-03-19
Also published as: AU8436791A; CA2073050A1; JPH05502456A; PT98869A; DE4028139A1; AU658148B2; TW197951B; EP0504339A1; KR920702235A; MX9100967A

Abstract

The use is disclosed of radioactive metal ion complexes with all-cis-1,3,5-triamino-2,4,6-trihydroxy-cyclohexane, their derivates, salts or quaternary ammonium salts for X-ray diagnosis and in tumor therapy, as well as for producing X-ray diagnostic and tumor therapy agents.

Description

Use of the complexes of radioactive metal ions with all-cis-1,3,5-triamino-2,4,6-cyclohexanetriol and its derivatives for

X-ray diagnostic purposes and in tumor therapy and for the production of agents for X-ray diagnostic purposes and for tumor therapy. The invention relates to the use of complexes of all-cis-1,3,5-

Triamino-2,4,6-cyclohexanetriol and its derivatives with radioactive ions

Metal isotopes, for X-ray diagnostic purposes and in tumor therapy and for the production of agents for X-ray diagnostic purposes and for tumor therapy.

The use of metal complexes for therapeutic or diagnostic

Application is in Inorganic Chemistry in Biology and Medicine, ACS

Symposium Series 140 ,. American Chemical Society, Washington, D.C. (1980),

Pages 91 to 101, 103 to 119 and 121 to 140 described. Complexes of radioactive isotopes have also been used for pharmaceutical purposes, in particular for diagnostic purposes. They can be used to display various body organs. A disadvantage of the previous display methods is the limited replaceability for certain areas of application.

For example, an X-ray representation of the human and animal heart was not possible.

The object of the invention is to provide new possibilities for X-ray diagnostic imaging and for tumor treatment.

It has been shown that this problem can be solved by

Use of complexes of radioactive metals, in particular radioactive metal cations with all-cis, 1,3,5-triamino-2,4,6-trihydroxycyclohexane and its derivatives, for X-ray diagnostic purposes and in tumor therapy or for the production of agents for X-ray diagnostic purposes and for tumor therapy.

The complexes used according to the invention are preferably radioactive

Underlying metal isotopes, which are α, ß, or γ emitters. Examples of radioactive isotopes that can be used are those of

Technetium, iron, gallium, ruthenium, indium, dysprosium, samarium, tullium, yterbium and yttrium. Specific examples are ions of the following isotopes:

⁵⁹ Fe, ⁶² Cu, ⁶⁷ Ga, ⁶⁸ Ga, ⁷² Ga, ⁹⁵ Ru, ⁹⁷ Ru, ⁹⁹ mTc, ^99m Rh, ¹¹¹ In, ¹¹³ In, ¹⁵⁷ Dy,

¹⁵³ Sm, ¹⁶⁷ Tm, ¹⁶⁹ Yb, ¹⁸² Re and ¹⁸⁶ Re. The ligands used for the complexes according to the invention are all-cis- 1,3,5-triamino-2,4,6-cyclohexanetriol and its derivatives, as they are

for example in EP-PS 0 190 676 (US-PS 4 794 197) and in Helv.

Chim. Acta, Vol 73, 1990, pages 97-105. They are manufactured using the methods specified there.

All-cis-1,3,5-triamino-2,4,6-trihydroxycyclohexane or its derivatives of the general formula (I)

I.

in question, wherein the symbols R ₁ , R ₂ , R ₃ , R ₄ , R ₅ or R _{6 are the} same or different and stand for hydrogen atoms, alkyl groups or -CO- alkyl groups, the alkyl in the alkyl or -CO -Alkylgruppen has 1 to 18 carbon atoms, and wherein the alkyl and -CO- alkyl groups can each independently contain one or more, identical or different functional groups, and their salts with inorganic or organic pharmacologically common acids and their quaternary ammonium salts of the general formula (II), (IIa) or (Ilb)

II I Ia Ilb with pharmaceutically acceptable anions, wherein R ₁ to R ₆ and R ₇ , R ₈ and

R ₉ independently of one another are the unsubstituted or substituted alkyl groups or -CO-alkyl groups defined above or all-cis- 1,3,5-tri-amino-2,4,6-trihydroxycyclohexane derivatives of the general formula

wherein the substituents on the nitrogen atoms are independently H or CH ₃ and preferably all CH ₃ .

Examples of the radicals R ₁ to R ₆ in the ligands are alkyl or —CO— alkyl groups which are one or more, identical or different

may contain functional groups. Of the symbols R ₁ to R _{9, there} is preferably at most one symbol on each nitrogen atom

secondary or tertiary alkyl substituent or a -CO-alkyl group. The other symbols preferably represent hydrogen or methyl.

A preferred group of ligands are those in which all residues

R ₁ to R ₆ or R ₁ to R _{9 are} methyl or -COCH ₃ groups.

The alkyl groups or —CO-alkyl groups of the substituents R ₁ to R ₆ or R ₁ to R ₉ preferably have one or more functional groups which can coordinate with metal cations. Examples of functional groups on the alkyl groups or -CO-alkyl groups are -OH, -COOH or salts thereof, -CONH ₂ , -CON (OH) R, where R represents an alkyl group having 1 to 6, in particular 1 to 4, carbon atoms , -OPO ₃ H ₂ and their salts, -PO ₃ H ₂ or their salts, -SR (in which R has the above meaning), and the esters of the above acids, -CN,

and or

and / or their salts.

Preferred examples are those of the ligands of the formulas (I), (II), (Ila) or (Ilb), in which one or more or all of the symbols R ₁ to R ₉ independently of one another have the following meaning:

where n is 1, 2 or 3, and their salts; as well as those in which one or more or all of the symbols R ₁ to R ₉ independently of one another have the following meaning:

where n is 1, 2 or 3, and their salts.

A particularly preferred group of ligands are those of the formula (I), (II), (Ila) and ((Ilb), in which at least one of the radicals R ₁ to R ₆ and in particular in each case the radicals R ₁ , R ₃ and R ₅ represent the group - (CH ₂ ) _n -COOH, in which n has the meaning from 1 to 3 and particularly preferably from 2. The remaining substituents R ₂ , R ₄ , R ₆ and R ₇ to R ₉ are preferably Hydrogen or CH ₃ , these remaining substituents preferably being the same Such compounds of the formula (I) are particularly well suited for tumor therapy.

A particularly important group of ligands according to the invention

Complexes used are those that have positive charges, so that the complexes made from it are positively charged. Such complexes are particularly good, for example, for X-ray diagnostic purposes,

especially for X-ray diagnostic imaging of the warm-blooded heart

(Humans and animals). The positive charges can either be caused by the quaternary nitrogen atoms of the compounds of the formula (II), (Ila),

(Ilb) and (IV) may be due, or by substituents with positive

charged functional groups, which are attached to the ligands of the formula (I) or (III), but also to the ligands of the formulas (II), (Ila), (Ilb) and

(IV) can be. The positive charges can, for example, by

Protonize or be achieved by quaternizing existing nitrogen atoms. Compounds of the formulas are particularly favorable examples

(II), (Ila), (Ilb) and (IV), in which all substituents (in the formulas

(II), (Ila), (Ilb) the substituent groups R ₁ to R ₉ ) represent methyl groups. Examples of substituents with positively charged

Functional groups are alkyl groups with quaternary ammonium groups, such as trimethylamino groups or triethylamino groups, for example those

of the formula - (CH ₂ ) _n -NR ₃ , where n is 1 to 3 and in particular 2 and R

Represents alkyl having 1 to 4 carbon atoms, especially methyl.

Particularly preferred compounds of the formula (I) are those in which R ₁ =

R ₃ = R ₅ = -CH ₂ CH ₂ COOH and R ₂ = R ₄ = R ₆ = H; as well as those in which R ₁ = R ₃ =

R ₅ = -CH ₂ CH ₂ N (CH ₃ ) ₃ and R ₂ = R ₄ = R ₆ = H.

The complexes used according to the invention are easily formed from aqueous solutions or suspensions of salts, hydroxides or oxides of the respective

radioactive metals of interest and aqueous solutions or suspensions of the ligands. The aqueous solutions can be acidic, for example. In general, amounts are used for this purpose such that 2 mol of ligands of the formula (I), (II), (Ila), (Ilb) or 1 mol of the formula (III) or (IV) are present per 1 mol of metal ion. A slight excess of the ligand is preferably used.

For the production of technetium complexes, for example, from

Technetiumhexafluorid or Technetiumhexachlorid solutions are started, which can be reduced initially so that Technetium (IV) ions are present, whereupon an aqueous solution with excess ligand is then added. In the complexes used according to the invention, part of the radioactive

Metals or metal cations can be replaced by corresponding non-radioactive isotopes. For example, the complexes can be in the form of mixtures of

Complex radioactive metals or metal cations with non-radioactive

Metals or metal cations are present. The non-radioactive

Metals or metal cations and the radioactive metals or metal cations may be the same or different. For example, if radioactive iron isotopes are used, they can be present together with non-radioactive iron. In the manufacture of such preparations, for example

Complex formation from a solution of iron III ions mixed with

radioactive iron III ions occur.

The complexes used according to the invention can be obtained from the manufacturer or

Suppliers are prepared; however, they are often also in situ in the

X-ray laboratory or treatment laboratory from the prepared solutions

manufactured.

The resulting aqueous solutions of complexes can be used for

X-ray diagnostic purposes and for tumor therapy.

Special fields of application are X-ray tomography, positron

Emission tomography and in particular the representation of the human and animal heart, for which purpose positively charged complexes are preferably used. For diagnostic purposes, it is particularly favorable to choose such complexes that γ- or

Are spotlights; α-emitters are preferably used for combating tumors, for example combating cancer.

The complexes used according to the invention are used in the form of aqueous solutions; they can be used in various ways, particularly by intravenous administration.

Example 1:

Synthesis of all-cis-1,3-5-tris- (2'-aminopropionic acid) -2,4,6-cyclohexanetriol

1 mmol (0.324 g) of all-cis-1,3,5-triammonio-2,4,6-cyclohexane-triol sulfate were dissolved in a little water and placed on an ion exchange column (Dowex 1, X 4, 50/100 mesh, OH form) applied. It was eluted with water until the eluate reacted neutral. The basic fractions were concentrated on a rotary evaporator. To this solution of 1 mmole of triaminocyclohexanetriol was added 3 mmoles (3 ml of 1M aqueous solution) of acrylonitrile and under for a week

Nitrogen left in the dark. The intermediate all-cis-1,3,5-tris- (2'-aminopropionitrile) -2,4,6-cycloheκan-triol was obtained by concentrating on a rotary evaporator.

¹ H NMR (90 MHz, D ₂ O, pD 2): 2.97 ppm (t, 6H); 3.49 ppm (t, 6H), 3.60 ppm

(m, 3H), 4.54 ppm (m, 3H). ^{1 3} C NMR (62.9 MHz, D ₂ O, pD 3): 14.9 ppm, 40.6 ppm, 57.0 pm, 63.2 ppm, 117.4 ppm. pK values (approximate); 3.0, 4.5, 6.3.

The nitrile was hydrolyzed by refluxing in concentrated hydrochloric acid for 6 hours. The all-cis-1,3,5-tris (2'ammoniopropionic acid) -2,4,6-cyclohexanetriol trichloride was obtained by concentrating on a rotary evaporator.

This was taken up in a little water and neutralized with potassium hydroxide solution to pH about 4.5. The all-cis-1,3,5-tris (2'-aminopropionic acid) - 2,4,6-cyclohexanetriol failed. The product was filtered off, dissolved in dilute acid (pH <3) and precipitated again by neutralization, filtered off and finally dried under high vacuum. Micro elemental analysis:

Substance formula: C ₁₅ H ₂₇ N ₃ O ₉ MG: 393.39

C H N

calculated 45.80% 6.92% 10.68%

found 41, 50% 7.24% 9.31%

These values are compatible with 2.8 moles of water of crystallization per mole of product.

¹ H-NMR (200 MHz, D ₂ O, acidic): 2.95 ppm (t, 6H), 3.53 ppm (t, 6H), 3.72 ppm (m, 3H), 4.72 ppm ( m, 3H).

¹³ C NMR (200 MHz, D ₂ O, acidic); 32.8 ppm, 43.5 ppm, 59.6 ppm, 66.1 ppm, 176.9 ppm. pK values (approximate): 6.2, 8.2, 9.7.

Example 2:

Synthesis of 59 ferri-all-cis-1,3,5-tris (2 ^, -aminopropionic acid) -2,4,6-cyclohexanetriol

1 mmol of all-cis-1,3,5-tris (2'-aminopropionic acid) -2,4,6-cyclohexanetriol (from Example 1) were dissolved in about 20 ml of 0.1 M hydrochloric acid. 1 mmol Fe (III) (1 ml 1M FeCl ₃ with 2 mCi ⁵⁹ FeCl, (Amersham) in HCl 1 M) was added. The clear, yellowish solution was slowly mixed with KOH (about 1 M) and it turned darker yellow. At pH approx. 3 the product started to fail. Now alkali was added to pH about 4.5.

The precipitated 59 ferri-all-cis-1,3,5-tris (2'-aminopropionic acid) -2,4,6-cyclohexanetriol was suction filtered through a G4 filter and then purified by dissolving in 0.1 M HCl and precipitation at pH 4.5. The product was finally dried under high vacuum. Because of spectroscopic

The structural identity with the non-radioactive labeled preparation was determined.

The analysis of the non-radioactive labeled complex is based on the formula

Close C ₁₅ H ₂₄ N ₃ O ₉ Fe.

Example 3:

Synthesis of all-cis-1,3,5-trihydroxy-cyclohexane-2,4,6-tri-N-acetylamide

3.28 g (18.5 mmol) of the amine (from Example 1) were dissolved in 100 ml of methanol and 35 ml (370 mmol) of acetic anhydride were added. The mixture was stirred at room temperature overnight. Methanol and excess

Acetic anhydride was removed in vacuo. The product obtained was recrystallized from ethanol and dried in a high vacuum over phosphorus pentoxide for 24 hours. Yield 86% mp 300 ° C. Mi kroe lementarana lysis:

¹ H NMR (200 MHz; D ₂ O) 2.05 ppm (s; 3H); 3.97-4.04 (q; 2H)

^{1 3} C-NMR (200 MHz, D ₂ O) 176.4 ppm; 72.8 ppm; 53.5 ppm, 24.7 ppm

Solubility: good solubility in water

At game 4:

Synthesis of 59 ferri-all-cis-1,3,5-trihydroxycyclohexane-2,4,6-tri-N-acetylamide

1 mmol of al-cis-1,3,5-trihydroxy-cyclohexane-2,4,6-tri-N-acetyl-amide were dissolved in about 20 ml of 0.1 M hydrochloric acid. 0.5 mmol Fe (III) (0.5 ml 1M FeCl ₃ in HCl 1M) with 2mCi ⁵⁹ FeCl ₃ (Amersham) were added.

Now the hydrochloric acid and a further 8 equivalents of alkali Pro Fe were added in one go with the precisely calculated amount of KOH (1M) (pH> 8). Then the neutralization with HCl (1M) down to the pH 5.0 ± 0.2 region is carried out under pH meter control. The uncharged iron complex of the

Composition Fe (H ₃ L ₂ ) from (L = ligand). The complex is soluble in or above a buffer with a physiological pH.

The structural identity with the non-radioactively labeled preparation was determined on the basis of spectroscopic properties.

Analysis of the non-radioactive labeled complex suggests the formula FeC ₂₄ H ₃₉ N ₆ O ₁₂ 8H ₂ O. At game 5:

Synthesis of gadolinium all-cis-1,3,5-tris- (2'-aminopropionic acid) -2,4,6-oyclohexanetriol

1 mmol of all-cis-1,3,5-tris (2'-aminopropionic acid) -2,4,6-cyclohexanetriol from (Example 1) were added in approx. 8 ml of 1 mol / 1

Hydrochloric acid dissolved. 1 mmol of GdCl ₃ (100 mg of GdCl ₃ * 6H ₂ O / ml

HCl 1 mol / l) added. The pH of the clear, colorless solution was slowly adjusted to 4.5 with NaOH (approx. 2 mol / l). This resulted in a first turbidity at pH 3 and one at pH 4.5

Precipitation. The precipitated gadolinium-all-cis-1,3,5-tris (2'-aminopropionic acid) -2,4,6-cyclohexanetriol was replaced by a G4-

Nutsche sucked off and then rinsed with distilled water. The product was finally dried under high vacuum at 40 ° C.

Micro elemental analysis:

These values are compatible with 15 moles of water of crystallization per mole of product.

Claims

Use of the complexes of radioactive metal ions with all-cis-1,3,5-triamino-2,4,6-cyclohexanetriol and its derivatives for X-ray diagnostic purposes and in tumor therapy and for the production of agents for X-ray diagnostic purposes and for tumor therapy. Claims

1. Use of the complexes of radioactive metal ions with all-cis-1,3,5-triamino-2,4,6-trihydroxycyclohexane or its derivatives of the general formula (I)

as ligands in which the symbols R ₁ , R ₂ , R ₃ , R ₄ , R ₅ or R _{6 are} identical or different and represent hydrogen atoms, alkyl groups or -CO- alkyl groups, the alkyl in the alkyl or -CO- Alkyl groups has 1 to 18 carbon atoms, and wherein the alkyl and -CO-alkyl groups can each independently contain one or more, identical or different functional groups, and their salts with inorganic or organic pharmacologically common acids or their quaternary ammonium salts of the general formula ( II), (Ila) or (Ilb)

with pharmaceutically acceptable anions, wherein R ₁ to R ₆ and R ₇ , R ₈ and R ₉ independently of one another have the same definitions as defined above

mean unsubstituted or substituted alkyl groups or -CO- alkyl groups, or with all-cis-1,3,5-triamino-2,4,6-tri-hydroxycyclohexane derivatives of the general formula

wherein the substituents on the nitrogen atoms are independently H or CH ₃ , as ligands, for X-ray diagnostic purposes and in tumor therapy and for the preparation of agents for

X-ray diagnostic purposes and for tumor therapy.

2. Use according to claim 1, characterized in that it is

the radioactive metal ions are α, β or γ rays.

3. Use according to claim 1 or 2, characterized in that the radioactive metal ions are ions of radioactive isotopes of iron, copper, gallium, yttrium, ruthenium, technetium, rhodium, indium, dysprosium, samarium, thulium, ytterbium or rhenium acts.

4. Use according to one of claims 1 to 3, characterized in that positively charged complexes are used, in particular for

X-ray diagnostic representation of the heart and for the production of means therefor.

5. Use according to claim 4, characterized in that complexes with the andes of the formulas (II), (Ila), (Ilb) or (IV) after

Sprue

starts.

6. Use according to claim 5, characterized in that the radicals R ₁ to R ₉ in the formulas (II), (Ila) and (Ilb) are each methyl groups and that the substituents on the nitrogen atoms on the formula (IV) are methyl groups .

7. Use according to claim 5, characterized in that the radicals R ₁ to R ₆ in the formulas (II), (Ila) and (IIb) are methyl groups and the radicals R ₇ , R ₈ and R _{9 are} each H.

8. Use according to one of claims 1 to 4, characterized in that the alkyl groups or -CO-alkyl groups contain one or more functional groups which can coordinate with metal ions.

9. Use according to claim 4, characterized in that the ligands of the general formula (I) are used, wherein at least one of the radicals R ₁ , R ₃ and R _{5 has} the general formula

- (CH ₂ ) _n - N

(R) has ₃ , where n = 1, 2 or 3, in particular 2, R is alkyl with 1, 2, 3 or 4 carbon atoms and the radicals R ₂ , R ₄ and R _{6 are} hydrogen atoms or alkyl groups with 1 to 18 carbon atoms .

10. Use according to one of claims 1 to 3, characterized in that ligands of the general formula (I), (II), (Ila) or (Ilb) are used, in which the radicals R ₁ , R ₃ and R _{5 are} functional groups

having alkyl groups with the general formula

- (CH ₂ ) _n - COOH, are,

where n = 1, 2 or 3 and in particular 2, and the radicals R ₂ , R ₄ and R ₆ to R _{9 are} each H.

11. Use according to claim 10 for tumor therapy or for

Manufacture of agents for tumor therapy.

12. A process for the preparation of complexes of radioactive metal ions for X-ray diagnostic purposes and for tumor therapy, characterized in that an aqueous solution or dispersion of a derivative of the general formula (I), (II), (Ila), (Ilb), (III ) or (IV), as defined in claim 1 7, 8, 9 or 10, with an aqueous solution or dispersion of a salt, hydroxide or oxide of a radioactive metal.

13. The method according to claim 12, characterized in that a salt, hydroxide or oxide of a metal is used, which is an α, β or γ emitter.

14. The method according to claim 12 or 13, characterized in that one uses the salt, hydroxide or oxide of a metal, which is one or more radioactive isotopes of iron, copper, gallium,

Yttrium, ruthenium, technetium, rhodium, indium, dysprosium, samarium, thulium, ytterbium or rhenium.

15. The method according to any one of claims 12 to 14, characterized in that for the preparation of an agent suitable for X-ray diagnostic purposes or for tumor therapy, the aqueous solution obtained according to one of claims 12 to 14, and this

optionally with customary pharmaceutical additives.