EP1011737A2

EP1011737A2 - Method for treating proliferative diseases by therapy

Info

Publication number: EP1011737A2
Application number: EP98954267A
Authority: EP
Inventors: Ludger Dinkelborg; Christoph-Stephan Hilger; Dieter Heldmann; Friedhelm Blume
Original assignee: Schering AG
Current assignee: Bayer Pharma AG
Priority date: 1997-09-18
Filing date: 1998-09-10
Publication date: 2000-06-28
Also published as: CA2301887A1; AU1145999A; WO1999013920A3; JP2001516730A; AU745908B2; WO1999013920A2

Abstract

The invention relates to a method for treating proliferative diseases by therapy, characterised in that an application catheter is first placed at the site of the lesion and a radioactive substance is applied locally. The catheter is then removed, leaving the radioactive substance at the site of the lesion. The invention also relates to the use of bis-amine-oxime derivative, N2S2 complex derivatives and radioactively marked colloidal solutions for producing agents which are applied locally in proliferative disease therapy.

Description

Method of therapeutic treatment of proliferative

Diseases

The invention is in the field of the therapy of proliferative diseases and in particular the therapy of vascular diseases such as atherosclerosis.

Ionizing radiation is known to inhibit cell proliferation. A variety of neoplastic and non-neoplastic diseases have been treated in this way (Fletcher, Textbook of Radiotherapy, Philadelphia, P.A: Lea and Febiger, 1980, Hall, Radiobiology for the Radiologist, Philadelphia, P.A: Lippincott, 1988).

Attempts have also been made to treat atherosclerotic diseases using these methods. Atherosclerosis is an inflammatory, fibroproliferative

Disease responsible for 50% of all deaths in the United States, Europe and Japan (Ross 1993, Nature 362: 801-809). With its peripheral characteristics it threatens the preservation of the extremities, with its coronary manifestation there is a risk of fatal heart attack and with supra-aortic involvement there is a risk of stroke.

Atherosclerosis is currently treated in different ways. In addition to the conservative measures (e.g. lowering the cholesterol level in the blood) and the bypass operation, the mechanical dilatation (angioplasty) and the intravascular removal of atheromatous tissue (atherectomy) of narrowed segments in the peripheral arteries and the coronary arteries have also been shown to be Alternative established in everyday clinical practice.

However, as explained below, the methods mentioned have a number of disadvantages.

The value of mechanically recanalizing procedures is acutely impaired by vascular occlusions as a result of vascular tears and dissections as well as acute thromboses (Sigwart et al. 1987, N. Engl. J. Med. 316: 701-706). Long-term success is endangered by the recurrence of restrictions (restenoses). The CAVEAT study showed that the restenosis rate of 1012 patients six months after the intervention was 50% in coronary atherectomy and even 57% in coronary angioplasty (Topol et al. 1993, N. Engl. J. Med. 329: 221 -227). Furthermore, abrupt vascular occlusions occurred in 7% of atherectomy patients and in 3% of angioplasty patients. Nicolini and Pepine (1992, Endovascular Surgery 72: 919-940) report one Restenosis rate between 35 and 40% and an acute closure rate of 4% after angioplasty

Various techniques have been developed to counter these complications. These include the implantation of metallic endoprostheses (stents), (Sigwart et al 1987, N Engl J Med 316 701-706, Strecker et al, 1990, Radiology 175 97-102) Large-caliber arteries, e.g. in occlusions in the pelvic axis, have already been given the status of a primary therapeutic modality.The use of stents in the femoral arteries, on the other hand, has shown disappointing results with a primary open rate of 49% and a reocclusion rate of 43% (Sapoval et al, 1992 , Radiology 184 833-839) Similar unsatisfactory results have been achieved with previously available stents in the coronary arteries (Kavas et al 1992, J Am Coll Cardiol 20 467-474)

To date, all previous pharmacological and mechanical interventions have not been able to prevent restenosis (Muller et al 1992, J Am Coll Cardiol 19 418-432, Popma et al 1991, Circulation 84 14226-1436)

The cause of the restenoses that often occur after mechanical interventions is believed to be that the interventions induce proliferation and migration of smooth muscle cells in the vascular wall. This leads to neointimal hyperplasia and the observed restenoses in the treated vascular sections (Cascells 1992, Circulation 86, 723-729 , Hanke et al 1990, Circ Res 67, 651-659, Ross 1993, Nature 362, 801-809)

An alternative method for the treatment of atherosclerotic diseases uses ionizing radiation. However, the use of external ionizing radiation on restenosis has the disadvantage that the radiation dose is not limited to the desired location during application, but rather surrounding (healthy) tissue Exposure to radiation is also undesirable. So far, various studies have shown little promise (Gellmann et al 1991, Circulation 84 Suppl II 46A-59A, Schwartz et al 1992, J Am Coll Cardiol 19 1106-1113)

These disadvantages, which occur when using external radiation sources, can be overcome if gamma radiation, e.g. via a catheter, is brought directly to the vascular areas with restenosis.This form of application with Iridium-192 means that a high radiation dose of 20 Gy is applied to the restenosis focus Some papers report the almost complete prevention of restenosis after this intervention (Wiedermann et al 1994, Am J Physiol 267 H125-H132, Böttcher et al 1994, Int J Radiation Oncology Biol Phys 29 183-186, Wiedermann et al 1994, J Am Coll Cardiol 23 1491-1498, Liermann et al 1994, Cardiovasc Intervent Radiol 17 12-16) Disadvantage of this method, however, is that the radiation dose of 20 Gy applied is very high. Because the lesions are irregularly distributed on the wall of the vessel, one Uniform application of a defined dose using this technique is not possible. Furthermore, treatment of large-caliber vessels is not possible because the dose that can be applied is insufficient due to the drop in dose from the iridium source

Another possibility of inhibiting restenosis is the implantation of P-32-doped stents (Fischell et al Stents III, Development, Indications and Future, Konstanz Kollath and Liermann, 1995). An activity of 0.2 kfiq was sufficient in this work P-32 per centimeter of stent length (corresponds to a radiation dose of 0.25 Gy) in order to achieve a maximum inhibition of the smooth vascular muscle cells in vitro. It could be shown that not only γ but also ß emitters prevent the proliferation of smooth muscle cells The advantage of this method is that the radiation dose applied is significantly lower than with all the interventions mentioned so far. At this low dose, the endothelial cells lining the vascular bed are not damaged (Fischell et al Stents III, Development, Indications and Future, Konstanz Kollath and Liermann, 1995) However, this form of intervention is only possible once, namely when positioning the stent he is only limited to such interventions in which stents are used. The restenosis that occurs in the much more frequently used interventions such as atherectomies and angioplasties cannot be treated with this method. Due to the short range of the ß radiation, the entire lesion cannot be given a uniform energy dose to administer

In addition to radiation therapy, a number of other therapeutic strategies for inhibiting neointimal hyperplasias (restenoses) are also used. These include classic drugs for restenosis suppression such as antithrombotics, antiplatelet agents, calcium antagonists, anti-inflammatory and anti-proliferative substances, but also gene therapy approaches Inhibition of growth stimulators, for example by antisense oligonucleotides, or the amplification of inhibiting factors by expression vector plasmids and virus-mediated gene integration possible. Aptamer oligonucleotides can also be used for inhibition a wide variety of receptor-mediated processes that play a crucial role in restenosis.

Substances that have been administered under strictly controlled conditions as long-term therapy have been examined with great energy and care for years, because it was theoretically hoped for a reduction in the restenosis rate (Herrmann et al., 1993, Drugs 46: 18-52).

More than 50 controlled studies with different substance groups were carried out without clear evidence that the tested substances could seriously reduce the restenosis rate. This also applies to local application, in which the substances are brought to the desired site of action via special balloon catheters. However, it has been shown that the substances used hitherto are washed out of the vessel wall too quickly in order to be therapeutically effective. In addition, these pressure-mediated liquid injections induce additional changes in the vessel wall, which even promote restenosis.

The object of the present invention was therefore to develop a method for the therapy of proliferative diseases which overcomes the disadvantages of the previously known therapy methods.

This object is achieved by the present invention.

A method for the therapeutic treatment of proliferative diseases has been developed, which is characterized in that first an application catheter is placed at the site of the lesion and a radioactive substance is applied locally via the catheter, then the catheter is removed again and the radioactive substance at the site of the Lesion remains.

Due to the fact that radioactive substances are targeted to the wall of a blood vessel via an application catheter and remain there, the concentration of the radionuclide lasts long enough to inhibit the proliferation of the cells and thus restenosis.

The method according to the invention has some essential advantages compared to the known therapy methods. In comparison to a large number of examined compounds from different classes, the local application leads to certain Substances and with certain catheters to a surprisingly high radioactive dose at the desired, pathologically changed location. This procedure leads to a high effective radiation dose with low systemic exposure. The radioactive substances have a long residence time at the application site, which leads to a high effective dose on site are mainly and evenly distributed in the pathological region. The unbound radioactive substances are quickly eliminated

Because certain radioactive substances, which are described in more detail below, get into the wall of the atherosclerotically modified vessels, not only the intimal cells facing the lumen, but also those of the media and adventitia are prevented from proliferating. The proportion of the dose applied that passes through the cell membrane leads to a high radiation dose, which is effective close to the cell nucleus

Because of the sensitivity of proliferating cells to ionizing radiation, the method according to the invention is not only suitable for the therapy of atherosclerotic diseases, but also for the therapy of other proliferative diseases, such as tumor diseases

Suitable radioactive substances are those which have a sufficiently high lipophilicity to adhere to the plaque. For example, radioactively labeled metal complexes are suitable, such as metal complexes of bis-amine oxime derivatives of the general formula I

wherein n = 0 - 3 and the radicals R to R ^ are the same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic C 1 -C 8 -oo-alkyl, alkenyl, alkynyl, Aryl, alkylaryl and / or arylalkyl radical, which may optionally contain fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, Aldehyde or alkoxy groups with up to 30 carbon atoms is substituted and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and wherein the radicals R ^ and R ^, R ⁴ and R ^ and R ° and R 'together can optionally represent an oxygen atom. Together with a radionuclide, these compounds form a metal complex which is then used for local application in the therapy of proliferative diseases.

The metal complexes of the N2S2 derivatives of the general formulas II and III are also suitable

wherein R ^ to R- ^ 2 are the same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic Ci -Ci rjo- alkyl, alkenyl, alkynyl, aryl, - Alkylaryl and / or arylalkyl radicals, which optionally have fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups up to 30 carbon atoms is substituted and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and the radicals R * and R ⁱ² , R ¹³ and R * ⁴ , R ¹⁵ and R * ⁶ and Rl and Rl8 together can optionally represent an oxygen atom and n, m and p independently of one another are 1 or 2. Other suitable compounds which are suitable for local therapy after complexing with suitable radioisotopes are tetrofosmin, sestamibi and furifosmin derivatives. ^99m Tc-Tetrofosmin is available under the trade name Myoview ™ from Amersham, ^99m Tc-Sestamibi is sold under the trade name Cardiolite® from DuPont, and ^99m Tc-Furifosmin is under the trade name TechneScan Q-12 from Mallinckrodt Medical to acquire.

All of these compounds together with a radionuclide form a metal complex which can then be used for local application in the therapy of proliferative diseases.

To form a metal complex, radionuclides can be introduced which are alpha, beta and / or Gam a emitters, positron emitters, Auger electron emitters and fluorescence emitters, with β and combined β / γ emitters for therapeutic Purposes are preferred.

Corresponding radionuclides are known to the person skilled in the art. The radionuclides of the elements of atomic numbers 27, 29 - 32, 37 - 39, 42 - 51, 62, 64, 70, 75, 77, 82 or 83 may be mentioned as examples.

The nuclides mτ ⁹⁹ _c, ^L86 R _e, ¹⁸⁸ Re, ⁶⁷ Cu, ⁹⁰ Y, and ¹⁰⁷ Ag are preferred which nuclides are especially preferred ¹⁸⁶ Re, ¹⁸⁸ Re, ⁶⁷ Cu.

The preparation of the bis-amine oxime derivatives is described in US Pat. Nos. 5,506,345 and 5,387,692; the production of the N2S2 derivatives is described in US Pat. No. 5,279,81 1.

The production of tetrofosmin derivatives is described in European patent application EP 303 374, the production of furifosmin derivatives is described in US Pat. No. 5, 112,595. Sestamibi derivatives and their preparation are described in international patent application WO 89/02433.

Other suitable metal complexes have ligands derived from ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA) or a macrocyclic compound such as e.g. Tetraazacyclododecan are derived. The making of this

Compounds are known to the person skilled in the art and are also described in detail in the examples below. Other suitable ligands are, for example, porphyrin derivatives, as described, for example, in DE 42 32 925 AI and DE 43 05 523 AI. Metal complexes suitable for the process according to the invention can also be prepared from these ligands using radionuclides

Radioactive thallium compounds of the isotopes ²⁰¹ T1, ²⁰⁷ T1, ²⁰⁹ T1 and ²¹⁰ T1 are also suitable; ²⁰¹ T1C1 is particularly suitable

Radioactive labeled colloidal solutions are also ideal for the treatment of proliferative diseases and especially for local application

Suitable colloidal solutions are the tin colloids described in the examples. Tin colloids are particularly suitable, which can be produced with the aid of a kit from Amersham ("Amerscan tin colloid ( ^99m Tc) - marking kit for liver scintigraphy"). Other suitable colloids are e.g. B radioactive gold sol ( ¹⁹⁸ Au colloid) and radioactively labeled sulfur colloid as well as other physiologically acceptable, radioactive colloidal solutions

Suitable radionuclides for radioactive labeling of the colloidal solutions are known to the person skilled in the art. The radionuclides of the elements Ag, As, At, Au, Ba, Bi, Br, C, Co, Cr, Cu, F, Fe, Ga, Gd, Hg may be mentioned as examples , Ho, I, In, Ir, Lu, Mn, N, O, P, Pb, Pd, Pm, Re, Rh, Ru, Sb, Sc, Se, Sm, Sn, Tb, Tc or Y

The nuclides ^99m Tc, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁶⁷ Cu, ⁹⁰ Y, ¹⁵ Sm, ¹⁶⁰ Tb, ¹⁶² Tb, ¹⁹⁸ Au and ¹⁰⁷ Ag are preferred

The colloidal solutions are usually prepared by means of a redox reaction or by changing the pH in an aqueous or alcoholic solution in the presence of a radioactive salt. The colloid can be formed in the presence of a stabilizer or can be added later with a surfactant or other stabilizing amphiphilic Substance added Other manufacturing methods for suitable colloidal solutions are electrochemical methods such as those described, for example, by MT Reetz et al in Angew Chem 1995, Vol 107, p. 2461 ff. The tin colloids are prepared in the examples below and in the instructions for use of the labeling kit described by Amersham The production of a gold colloid for diagnostic purposes is described in the patent specification DE 24 20 531 C3 The size of the particles formed is in the range between 5 and 1000 nm, in the case of the tin colloid between 300 and 600 nm.

The catheters outlined in FIG. 3 can be used as catheters which are suitable for the local application of the substances according to the invention. Multi-chamber balloon catheters (such as Dispatch ™, SciMed) and microperforated balloon catheters are particularly suitable.

In the examples below, the method is described in animal experiments. The preparation of some compounds suitable for use in this therapeutic method is also described. In Examples 1 to 5, the process is carried out with "rnTc-labeled HMPAO, the ligand HMPAO having the following structure:

(see also Radiopharmaceuticals, Chemistry and Pharmacology, edited by Adrian D. Nunn, 1992, page 53).

example 1

Local application of "mTc-HMPAO

The experimental animal, a white New Zealand rabbit (internal animal identification number: 1708, male, 3.7 kg body weight) was prepared 4 weeks before the actual application experiment as follows:

In anesthesia (Rompun / Ketavet 1: 2, 1 ml / kg body weight, i.m. administration) the endothelium was damaged with a 2F Fogarthy balloon catheter in the arteria carotis dextra (balloon nudation). The animal then received a special diet with an addition of 0.2% cholesterol. As a result of this pretreatment, the test animal develops an atherosclerotic lesion at the balloon-diffused site.

The local application of the FLMPAO marked with Technetium 99m takes place on the anesthetized test animal (anesthetic type see above) via a coronary perfusion / infusion catheter (Dispatch 3.0, Xtra slippery coating, manufacturer: Boston Scientific Corporation, Ratingen) directly at the lesion in the carotid artery. The radioactive dose of 0.48 mCi (= 17.76 MBq) was applied in a volume of 0.85 ml.

During the entire experiment, the test animal was placed under a gamma camera (Elscint SP4 HR) to measure the distribution of radioactivity in the body. The activity at the lesion is related to the total activity (measured in the animal at that point in time). In this experimental animal there were: 55.38% of the dose at the lesion 5 minutes after application

4 hours after application 46.78% of the dose at the lesion 24 hours after application 21.45% of the dose at the lesion

Example 2

Local application of ^99m Tc-HMPAO

The experimental animal, a white New Zealand rabbit (internal animal identification number: 1856, male, 3.3 kg body weight) was prepared 4 weeks before the actual application experiment as follows:

In anesthesia (Rompun / Ketavet 1: 2, 1 ml / kg body weight, in-dose) the endothelium was damaged with a 2F Fogarthy balloon catheter in the carotid artery (Balloon nudation) The animal then received a special diet with an addition of 0 2% cholesterol. With this pretreatment, the experimental animal developed an atherosclerotic lesion at the balloon nudated site

The local application of the HMPAO marked with Technetium 99m takes place on the anesthetized test animal (anesthetic type see above) via a coronary perfusion / infusion catheter (Dispatch 3 0, Xtra slippery coating, manufacturer Boston Scientific Corporation, Ratingen) directly at the lesion in the A carotis die radioactive Dose of 1.91 mCi (= 70 67 MBq) was applied in a volume of 1 0 ml (rinsing with 0 3 ml of physiological saline solution)

During the entire experiment, the test animal is under a gamma camera (Elscint SP4 HR) to measure the distribution of radioactivity in the body. The activity at the lesion is related to the total activity (measured in the animal at this point in time) Experimental animals were at the lesion 5 minutes after application 40 40 74% of the dose

4 hours after application 35 13% of the dose at the lesion 24 hours after application 23 23% of the dose at the lesion

Example 3

Local application of 99mχ _c .HMPAO

The test animal is a white New Zealand rabbit (internal animal identification number 1584, male, 3.4 kg body weight)

In anesthesia (Rompun / Ketavet 1.2, 1 ml / kg body weight, in the dose), the endothelium was damaged with a balloon catheter in the infrared aorta (balloon nudation). TechMPium 99m-labeled HMPAO was then applied to the test animal over a period of 5 minutes using a microperforated balloon catheter (4 mm Match-35 PTA, Schneider, FRG) applied The radioactive dose of 0 64 mCi (= 23 68 MBq) was applied in a volume of 1 ml

During the entire experiment, the test animal is under a gamma camera (Elscint SP4 HR) to measure the distribution of radioactivity in the body. The activity at the lesion is related to the total activity (measured in the animal at this point in time) There were experimental animals

5 minutes after application 38 45% of the dose to the lesion 4 hours after application 35 64% of the dose to the lesion 24 hours after application 16 63% of the dose to the lesion Example 4

Local application of ^99m Tc-HMPAO

The test animal was a white New Zealand rabbit (internal animal identification number: 1587, male, 3.5 kg body weight).

In anesthesia (Rompun / Ketavet 1: 2, 1 ml / kg body weight, i.m. administration), the endothelium was damaged with a balloon catheter in the infrared aorta (balloonudation). Subsequently, technetium 99m-labeled HMPAO was applied to the test animal over a period of 5 minutes using a microperforated balloon catheter (4 mm Match-35 PTA, from Schneider, FRG). The radioactive dose of 1.18 mCi (= 43.66 MBq) was applied in a volume of 1 ml. During the entire experiment, the test animal was placed under a gamma camera (Elscint SP4 HR) to measure the distribution of radioactivity in the body. The activity at the lesion is related to the total activity (measured in the animal at that point in time). In this experimental animal there were:

5 minutes after application 37.06% of the dose at the lesion 4 hours after application 32.03% of the dose at the lesion

24 hours after application 20.01% of the dose at the lesion

Example 5

Local application of "mTc-HMPAO

The test animal was a white New Zealand rabbit (internal animal identification number: 1586, male, 3.3 kg body weight).

In anesthesia (Rompun / Ketavet 1: 2, 1 ml / kg body weight, i.m. administration), the endothelium was damaged with a balloon catheter in the infrared aorta (balloonudation). Subsequently, technetium 99m-labeled HMPAO was applied to the test animal over a period of 5 minutes using a microperforated balloon catheter (4 mm Match-35 PTA, from Schneider, FRG). The radioactive dose of 0.45 mCi (= 16.65 MBq) was applied in a volume of 1 ml.

During the entire experiment, the test animal was placed under a gamma camera (Elscint SP4 HR) to measure the distribution of radioactivity in the body. The activity at the lesion is related to the total activity (measured in the animal at that point in time). In this experimental animal there were: 5 minutes after application 45.56% of the dose at the lesion 4 hours after application 36.39% of the dose at the lesion 24 hours after application 15.24% of the dose at the lesion

Example 6

Preparation of l- {3- [N- (2-methoxyethyl) octadecylsulfamoyl] -2-hydroxypropyl} -4,7,10-tetraaza-cyclododecane, yttrium-90 complex

5 mg l- {3- [N- (2-methoxyethyl) octadecylsulfamoyl] -2-hydroxypropyl} -4,7, 10-tetraazacyclodo-decane (manufactured according to DE 4340809.5) are dissolved in 500 μl dimethyl sulfoxide and 50 μl 0.1 M sodium acetate buffer (pH = 4.0) dissolved. After adding 37 MBq of yttrium-90-trichloride solution, the reaction mixture is heated at 100 ° C. for 10 min. The Y-90 complex solution prepared in this way can be used without further purification.

Example 7

a) Preparation of N, N'-bisundecyl-diethylene-triamine-pentaacetic acid diamide

3.57 g (10 mmol) of diethylene-triamine-pentaacetic acid bisanhydride are suspended together with 4.05 g (40 mmol) of triethylamine in 100 ml of absolute dimethylformamide. A solution of 3.42 g (20 mmol) of undecylamine, dissolved in 50 ml of absolute dichloromethane, is then added dropwise to the reaction mixture at room temperature. The reaction mixture is stirred for 6 h at room temperature, filtered and evaporated under a fine vacuum. The residue is dissolved three times in 100 ml of dimethylformamide and each evaporated under a fine vacuum. 50 ml of absolute diethyl ether are poured over the foamy reaction product and the mixture is stirred overnight. It is filtered and dried in a fine vacuum.

Yield: 6.3 g (90%), white powder.

Elemental analysis: Calc .: C 61.77 H 9.94 N 10.01 O 18.86

Found: C 61.52 H 9.63 N 9.91 O b) Preparation of N, N'-bisundecyl-diethylenetriamine-pentaacetic acid diamide, yttrium-90 complex

5 mg of N.N'-bisundecyl-diethylenetriamine-pentaacetic acid diamide (Example 7a) are dissolved in 500 μl of dimethyl sulfoxide and 50 μl of 0.1 M sodium acetate buffer (pH = 4.0). After adding 37 MBq of yttrium-90-trichloride solution, the reaction mixture is left to stand at room temperature for 10 min. The Y-90 complex solution prepared in this way can be used without further purification.

Example 8

a) Preparation of N-benzyloxycarbonyl-glycyl-N'-undecyl-glycinamide

3.63 g (10 mmol) of N-benzyloxycarbonyl-glycyl-glycine-N-hydroxysuccinimide ester and 1.71 g (10 mmol) of undecylamine are dissolved in 100 ml of absolute dichloromethane. The reaction mixture is stirred for 6 hours at room temperature. The mixture is then diluted with 100 ml of dichloromethane and the organic phase is washed twice with 50 ml of saturated sodium bicarbonate solution and once with 50 ml of water. It is dried over magnesium sulfate and the solvent is evaporated off in vacuo. The crude product is purified by chromatography on silica gel (eluent: dichloromethane / methanol 95: 5).

Yield: 3.8 g (90.6%), white powder.

Elemental analysis: Calc .: C 65.84 H 8.89 N 10.01 O 15.25 Found: C 65.71 H 9.02 N 10.10 O

b) Preparation of glycyl-N'-undecyl-glycinamide

3 g (7.15 mmol) of N-benzyloxycarbonyl-glycyl-N'-undecyl-glycinamide (Example 8a) are dissolved in 100 ml of absolute ethanol. After adding 300 mg of palladium on carbon (10%), the mixture is hydrogenated at room temperature (1 atm of hydrogen) for 2 h. It is filtered and evaporated in vacuo. The resulting amine is used for the subsequent reaction without further purification.

Yield: 1.92 g (94.1%), white foam. Elemental analysis Ber C 63 12 H 10 95 N 14 72 O 11 21

Gef C 63 03 H 11 04 N 14 57 O

c) Preparation of N- (S-acetyl-mercaptoacetyl) -glycyl-N'-undecyl-glycina id

285 4 mg (1 mmol) of glycyl-N'-undecyl-glycinamide (Example 8b) and 231 2 mg (1 mmol) of S-acetyl-mercapto-acetic acid-N-hydroxy-succinimide ester are dissolved together in 20 ml of absolute dichloromethane the reaction mixture for 6 h at room temperature, then it is diluted with 20 ml of dichloromethane, the organic phase is washed twice with 5 ml of semi-saturated sodium bicarbonate solution and once with 5 ml of water. It is dried over magnesium sulfate and the solvent is evaporated in vacuo. The crude product is purified by chromatography on silica gel ( Eluens dichloromethane / methanol 93 7) cleaned

Yield 362 mg (90 1%), white powder

EA Ber C 56 83 H 8 79 N 10 46 O 15 94 S 7 98

Gef C 56 67 H 8 93 N 10 18 O S 7 72

d) Preparation of N- (mercaptoacetyl) -glycyl-N'-undecyl-glycinamide

201 mg (0 5 mmol) of N- (S-acetyl-mercaptoacetyl-glycyl-N'-undecyl-glycinamide (Example 8c) are dissolved in 15 ml of absolute ethanol. Saturated with argon and an ammonia stream is passed through the for 30 min Solution The mixture is then evaporated and the residue is taken up in 20 ml of dichloromethane. The organic phase is shaken once with 2% aqueous citric acid and dried over sodium sulfate. The solvent is evaporated in vacuo and the residue is chromatographed on silica gel (eluent dichloromethane / methanol 9 l)

Yield 153 mg (85 1%), white powder

EA Ber C 56 79 H 9 25 N 11 69 O 13 35 S 8 92

Gef C 56 67 H 9 43 N 11 48 OS 8 71 e) Preparation of N- (mercaptoacetyl) -glycyl-N'-undecyl-glycinamide, Re-186 - complex

5 mg of N- (mercaptoacetyl) -glycyl-N'-undecyl-glycinamide (Example 8d) are dissolved in 800 μl of ethanol. After adding 5 mg disodium L-tartrate, 50 μl 0.1 M sodium hydrogen phosphate buffer (pH = 8.5), 37 MBq perrhenate and 10 μl tin dichloride dihydrate solution (5 mg SnCl2x2H ₂ O / lml 0.1M HC1) are added. The reaction mixture is heated at 60 ° C. for 5 min. The solution of the Re-186 complex of N- (mercaptoacetyl) -glycyl-N'-undecyl-glycinamide thus prepared can be used without further purification.

Example 9

Preparation of N, N'-bis [3,6,9,9-tetra (hydroxycarboxymethyl) -l-oxo-3,6,9-triaza-non-l-yl] mesoporphyrin-IX-13,17-dihydrazide , Y-90-

complex

5 mg of the N, N'-bis [3,6,9-tri (hydroxycarboxymethyl) -9- (ethoxycarboxymethyl) -l-oxo-3,6,9-triaza-non-l-yl] mesoporphyrin-IX- 13,17-dihydrazides (produced according to DE 42 32 925 AI, example la) are stirred in 5 ml 0.1M NaOH under an argon atmosphere for 3 hours at room temperature. After saponification of the bis-ethyl ester (TLC control), the pH is adjusted to 6 using glacial acetic acid and 37 MBq of yttrium-90-trichloride solution are added to the batch. The mixture is stirred for 15 minutes at room temperature. The HPLC analysis shows a 95% incorporation of the radioisotope.

Example 10

Preparation of 5,10,15,20-tetrakis [3- (carboxymethoxy) phenyl] porphyrin, yttrium 90 complex

2.0 mg of 5,10,15,20-tetrakis [3- (carboxymethoxy) phenyl] porphyrin (prepared according to DE 43 05 523 AI, Example 13a) are dissolved in 5 ml of acetic acid and with a hydrochloric acid solution of 1 , 0 mCi yttrium-90-chloride were added. The reaction mixture is autoclaved at 140 ° C. for one hour, the solvent is evaporated off in vacuo and the residue is taken up in 5 ml of water. By dropwise addition of aqueous sodium hydrogen carbonate pH 7.3 is set and the resulting red solution Filtered through a membrane filter By HPLC control of the filtrate, an incorporation rate of> 95% of the activity used in the porphyrin ligands can be determined

Example 11

Preparation of 5,10,15,20-tetrakis [3- (carboxymethoxy) phenyl] porphyrin, copper-67 complex

The preparation of the complex is described in DE 43 05 523 AI, example 14

Example 12

Production of a technetium-99m tin colloid

555 MBq sodium pertechnetate-99m in 2 ml 0 9% sodium chloride solution are mixed with 20 μl tin-II-chloride solution (5 mg tin-II-chloride dihydrate / 1 ml 0 01M HC1) at room temperature. After 10 minutes, dilute with 1 ml PBS buffer The solution obtained is slightly opalescent

Example 13

Preparation of a rhenium-186-tin colloid

37 MBq of sodium perrhenate-186 in 2 ml of 9% sodium chloride solution are mixed with 40 μl of tin-II-chloride solution (5 mg of tin-II-chloride dihydrate / 1 ml of 0 01M HC1) at room temperature. After 10 minutes, dilute with 1 ml PBS buffer The solution obtained is slightly opalescent

Example 14

Local application of a tin colloid

The test animal is a white New Zealand rabbit (internal animal identification number 1852, male, 3.5 kg body weight)

In anesthesia (Rompun / Ketavet 1 2, 1 ml / kg body weight, in the dose) the endothelium was damaged with a balloon catheter in the infrared aorta (balloon diffusion) The test animal was then tin-colloided over a period of 5 minutes using a microperforated match catheter (balloon catheter with a diameter of 5 mm, manufacturer Fa Schneider, Dusseldorf), which according to the kit from Amersham ("Amerscan tin colloid ( ^99m Tc) - marking kit for liver scintigraphy ") was produced, applied The radioactive dose of 0 4 mCi (= 14 8 MBq) was applied in a volume of 0.1 ml

During the entire experiment, the test animal was placed under a gamma camera (Elscint SP4 HR) to show the distribution of radioactivity in the body. In Fig. 1 the situation before application was shown in the upper part. The catheter containing the tin colloid was clearly visible Arrow shows the balloon of the catheter, which is located at the desired application site. In the lower part of the image, the same site is shown 1.5 hours after application and removal of the catheter. The amount of tin colloid remaining at the application site can be clearly seen

Example 15

Local application of a tin colloid

The test animal is a white New Zealand rabbit (internal animal identification number 1839, male, 3.7 kg body weight)

In anesthesia (Rompun / Ketavet 1 2, 1 ml / kg body weight, in the dose), the endothelium was damaged with a balloon catheter in the infrared aorta (balloonudation). The test animal was then catheterized over a period of 5 minutes with a microperforated match (balloon catheter with 5 mm diameter, manufacturer Fa Schneider, Dusseldorf) tin colloid, which was manufactured according to the kit from Amersham ("Amerscan tin colloid ^99m Tc - marking kit for liver scintigraphy"). The radioactive dose of 0 47 mCi (= 17 39 MBq) was applied in a volume of 0.1 ml

During the entire experiment, the test animal is under a gamma camera (Elscint SP4 HR) to show the distribution of radioactivity in the body. The situation before application is shown in the upper part of FIG. 2. The catheter that contains the tin colloid can be clearly seen. The arrow shows the balloon of the catheter that is located at the desired application site. In the lower part of the image, the same site is shown 1.5 hours after application and removal of the catheter Application site to recognize the remaining amount of tin colloid

Claims

claims

1. A method for the therapeutic treatment of proliferative diseases, characterized in that first an application catheter is placed at the site of the lesion and a radioactive substance is applied locally via the catheter, then the catheter is removed again and the radioactive substance remains at the site of the lesion.

2. A method for the therapeutic treatment of atherosclerotic diseases, characterized in that first an application catheter is placed at the site of the lesion and a radioactive substance is applied locally via the catheter, then the catheter is removed again and the radioactive substance remains at the site of the lesion.

3. The method according to claim 1 or 2, characterized in that the radioactive substance is a metal complex.

4. The method according to claim 1 or 2, characterized in that the radioactive substance is a metal complex whose ligand is a bis-amine oxime derivative of the general formula I,

wherein n = 0 - 3 and the radicals R * to R ^ are the same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic Cι -Cιoθ "lkyK -alkenyl, alkynyl, aryl -, -

Alkylaryl and / or arylalkyl radicals, which optionally have fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups up to 30 carbon atoms is substituted and / or optionally by one or several heteroatoms from the series N, P, As, O, S, Se are interrupted and / or substituted, and the radicals R ² and R ³ , R ⁴ and R ^ and R ^ and R ⁷ together optionally represent an oxygen atom can and whose central atom is a radionuclide of the elements of atomic numbers 27, 29 - 32, 37 - 39, 42 - 51, 62, 64, 70, 75, 77, 82 or 83.

5. The method according to claim 1 or 2, characterized in that the radioactive substance is a metal complex, the ligand of which is an N2S2 derivative of the general formula II,

wherein R ^ to R ² ^ are the same or different and each represents a hydrogen atom and / or an unbranched, branched, cyclic or polycyclic Ci -CioO "alkyl, alkenyl, alkynyl, aryl, alkylaryl and / or arylalkyl radical, optionally with fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups with up to 30 carbon atoms and / or is optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and where the radicals R ¹ and R ¹² , R ¹³ and R ¹⁴ , R ¹⁵ and R ¹⁶ and R ¹⁷ and R ¹⁸ together can optionally represent an oxygen atom and n, m and p independently of one another represent 1 or 2, and the central atom of which is a radionuclide of the elements of atomic numbers 27, 29 - 32, 37 - 39, 42-51, 62, 64, 70, 75, 77, 82 or 83.

6. The method according to claim 1 or 2, characterized in that the radioactive substance is a metal complex, the ligand of which is an N2S2 derivative of the general formula III,

wherein R ² ^ to R ^{32 are the} same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic CI -CI QQ- alkyl, alkenyl, alkynyl, aryl, alkylaryl - And / or arylalkyl radical, which optionally with fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups with up is substituted to 30 carbon atoms and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and the central atom of which is a radionuclide of the elements of atomic numbers 27, 29-32, 37-39, 42-51, 62, 64, 70, 75, 77, 82 or 83.

7. The method according to claim 4, 5 or 6, characterized in that the metal complex used contains a central atom which is selected from the group ^99m Tc, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁶⁷ Cu, ⁹⁰ Y and ¹⁰⁷ Ag.

8. The method according to claim 1 or 2, characterized in that the radioactive substance is a metal complex whose ligand is a porphyrin derivative.

9. The method according to claim 1 or 2, characterized in that the radioactive substance is a thallium compound of the isotopes ²⁰¹ T1, ²⁰⁷ T1, ²⁰⁹ T1 and ²¹⁰ T1.

10. The method according to claim 1 or 2, characterized in that the radioactive substance ^{201 is} T1C1.

Process according to claim 1 or 2, characterized in that the radioactive substance is a tetrofosmin derivative.

12. The method according to claim 1 or 2, characterized in that the radioactive substance is a sestamibi derivative.

13. The method according to claim 1 or 2, characterized in that the radioactive substance is a furifosmin derivative.

14. The method according to claim 1 or 2, characterized in that the radioactive substance is a colloidal solution with particle sizes between 5 and 1000 nm.

15. The method according to claim 1 or 2, characterized in that the radioactive substance is ^99m Tc-tin colloid or ¹⁸⁶ Re-tin colloid.

16. The method according to claim 1 or 2, characterized in that the catheter used is a microporous balloon catheter.

17. The method according to claim 1 or 2, characterized in that the catheter used is a multi-chamber balloon catheter.

18. Use of complexes whose ligand is a bis-amine oxime derivative of the general formula I.

wherein n = 0-3 and the radicals R ¹ to R ^{8 are the} same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic Ci -Ci QO- alkyl, alkenyl, alkynyl , Aryl, alkylaryl and / or arylalkyl radical, which may optionally contain fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy,

Aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups with up to 30 carbon atoms is substituted and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and where the radicals R ² and R ³ , R ⁴ and R ^ and R ^ and R ⁷ together may optionally represent an oxygen atom,

and whose central atom is a radionuclide of the elements of atomic numbers 27, 29 - 32, 37 - 39, 42 - 51, 62, 64, 70, 75, 77, 82 or 83, for the production of agents which are used in the therapy of proliferative diseases be applied locally.

19. Use of complexes whose ligand is an N2S2 derivative of the general formula II

where R ^ to R ⁽ ^ are the same or different and each for a hydrogen atom and / or for an unbranched, branched, cyclic or polycyclic CI -CI QO- alkyl, alkenyl, alkynyl, aryl, alkylaryl - and / or arylalkyl radical, which may optionally contain fluorine, chlorine, bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl,

Alkoxycarbonyl, amino, aldehyde or alkoxy groups with up to 30 carbon atoms is substituted and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se, and wherein the Radicals R ^{1 1} and R ¹² , R ¹³ and R ¹⁴ , R ¹⁵ and R ¹⁶ and Rl and Rl8 together may optionally represent an oxygen atom and n, m and p independently of one another are 1 or 2, and whose central atom is a radionuclide of the elements of atomic numbers 27, 29 - 32, 37 - 39, 42 - 51, 62, 64, 70, 75, 77, 82 or 83, for the production of agents which are used in the therapy of proliferative diseases be applied locally.

20. Use of complexes whose ligand is an N2S2 derivative of the general formula III

where R * to R ^{32 are the} same or different and each for one

Hydrogen atom and / or an unbranched, branched, cyclic or polycyclic C JC I QO "alkyl, alkenyl, alkynyl, aryl, alkylaryl and / or arylalkyl radical, optionally with fluorine, chlorine -, Bromine and / or iodine atoms and / or hydroxyl, oxo, carboxy, aminocarbonyl, alkoxycarbonyl, amino, aldehyde or alkoxy groups with up to 30

Is substituted and / or optionally interrupted and / or substituted by one or more heteroatoms from the series N, P, As, O, S, Se,

and its central atom is a radionuclide of the elements of atomic numbers 27, 29 - 32,

37-39, 42-51, 62, 64, 70, 75, 77, 82 or 83, for the preparation of agents which are applied locally in the therapy of proliferative diseases.

21. Use of compounds according to any one of claims 18 to 20, characterized in that the radionuclide is selected from the group "^ Tc, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁶⁷ Cu, ⁹⁰ Y and ¹⁰⁷ Ag.

22. Use of colloidal solutions for the preparation of agents for the therapy of proliferative diseases, characterized in that that the colloidal solution with a radionuclide of the elements Ag, As, At, Au, Ba, Bi, Br, C, Co, Cr, Cu, F, Fe, Ga, Gd, Hg, Ho, I, In, Ir, Lu , Mn, N, O, P, Pb, Pd, Pm, Re, Rh, Ru, Sb, Sc, Se, Sm, Sn, Tb, Tc or Y is marked.

23. Use of colloidal solutions according to claim 22, characterized in that the colloidal solution with a radionuclide selected from the group ^99m Tc, ¹⁸⁶ Re, ¹⁸⁸ Re, ⁶⁷ Cu, ⁹⁰ Y, ¹⁵³ Sm, ¹⁶⁰ Tb, ¹⁶² Tb, ¹⁹⁸ Au and ¹⁰⁷ Ag is marked.

24. Use of colloidal solutions according to claim 22, characterized in that the colloid is produced by a redox reaction in the presence of a radioactive salt.

25. Use of colloidal solutions according to claim 22, characterized in that the colloid is produced by changing the pH in an aqueous or alcoholic solution in the presence of a radioactive salt.

26. Use of colloidal solutions according to claim 22, characterized in that the particle size of the colloidal particles is between 5 and 1000 nm.

27. Use of colloidal solutions according to claim 22, characterized in that the particle size of the colloidal particles is between 300 and 600 nm.

28. Use of colloidal solutions according to claim 22, characterized in that the colloidal solution with the aid of surfactants or other amphiphilic

Substances is stabilized.

29. Use of radioactively labeled sulfur colloids for the production of agents for the therapy of proliferative diseases.