CA2048899A1 - Reducing chelating agents, their technetium and rhenium complexes, process for their production as well as their use in diagnosis and treatment - Google Patents

Abstract

Abstract of the Disclosure Compounds of formula I

(I) are provided, which have the capability of reducing pertechnetate without adding reducing agents to an oxidation state smaller than +7, to form, by groups Y and Z, stable complexes with the thus reduced technetium or rhenium isotopes and to accumulate selectively in certain tissues or lesions, which selectivity is optionally because of a compound coupled through a functional group contained in R1.
The technetium and rhenium complexes of the compounds of formula I, as well as their salts with organic and inorganic acids, can be used in in vivo diagnosis and treatment, e.g., in the treatment of tumors.

Description

RED~CING C~E~rING AGENTS, THEIR TEC~NETI~M AND R}IENI~M COMPLEXES, PROCESS FOR THEIR PROD~CTION
A8 WELL ~S T~EIR I~SE IN I)IAGNOSI~ AND TREATMENT

Backqround of the Invention Radioactive metal ions, usually bound to a complexing agent, have been used for in vivo diagnosis for some time.
of these, technetium-99m (Tc-99m), because of its almost ideal physical properties for these purposes -- good absorption of radiation in corresponding detection devices (gamma camera, SPECT devices) relative to a low absorption in the human organism and easy availability with a molybdenum/technetium generator -- is the radionuclide most often used in clinical nuclear medicine. Its short half-life of 6.02 hours guarantees an only slight exposure of the patient to gamma radiation, particularly since also the secondary product technetium-99 has only an insignificant residual radiation. But a drawbac~ of the technetium is its complicated and not yet completely known complex chemistry.
Technetium can be present in a number of oxidation states (+7 to -1), and the pharmacological properties can be greatly changed by changing the charge of a complex. It is therefore necessary to use complexes which bind the technetium in a defined oxidation state and to prevent redox :;:

2~4~9 reactions, which could lead to a redistribution of the pharmaceutical agent.
For organ- or tissue-specific diagnosis, it is necessary that the radiopharmaceutical agents be selectively S concentrated in the desired target organs or tissues and remain there for a while. This selectivity can be achieved, on the one hand, by the formation of complexes, which on their own show a specificity for certain tissues, or by coupling the technetium complexes to selecti~e substances, such as, e.g., monoclonal antibodies.
For labeling organ-specific substances with Tc~99m, the pertechnetate eluted from the nuclide generator first has to be converted to a lower oxidation state. In this reduced form, technetium forms more or less stable compounds with the selectively concentrated substances. The special problem of labeling with Tc-99m consists in the fact that normally tin(II) ions are present in the reaction solution as reducing agents. Tin(II) is thus far the only reducing agent which makes possible a quick and quantitative conversion of the pertechnetate at room temperature to a lower and thus reactive oxidation state. There, the added tin(II) salts have to be used in a high excess (about 100:1) relative to the pertechnetate. But the tin(II) and tin(IV) ions present after the reduction has been completed, in addition to the reduced Tc-99m, compete for the binding sites of the ligands, so that either the complexing agent again has to be used in excess relative to the tin, by which the specific activity is greatly reduced or unbound Tc-99m and tin as common colloid results in undesirable storage of radioactivity in other organs. In both cases, the diagnostic informative value is reduced.
This prcble~ can be avoided by the use of reducing ligands. In the production of diagnostic agents according - . ' .
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2 0 '~ 9 to this principle, a part of the added ligand excess acts as reducing agent for pertechnetate, which reduces technetium in an oxidation state lower than +7. In this way, reduced technetium species are then complexed by the excess of the unoxidized chelating agent. In this case, it is important to obtain stable complexes in a defined oxidation state for the technetium.
DeLearie et al. (L. A. deLearie, R. C. Haltiwanger, C.
G. Pierpont; J. Am. Chem. Soc. 111: 4324, 1989) showed that 3,5-di-tert-butylcatechols are suitable for reduction and chelation of Tc-99 (half~life: 212,000 years). The reduction took place by 24 hour-s of boiling in methanol.
For the labeling with the short-lived isotope of technetium (Tc-99m; half-life: 6 hours), however, only substances are usable as radiopharmaceutical agents which can be labeled quickly and gently also in the clinic and for which no subsequent purification after the labeling with, eOg., Tc-99m is necessary. The compounds described by DeLearie are thus unsuitable for the production of a clinically usable radiopharmaceutical agent.

Summary of the In~ention The present invention provides new reducing and tissue-specific chelating agents, as well as their stable technetium and rhenium complexes. Surprisingly, substances were ound which reduce and completely complex Tc-99m under mild conditions as well as quickly. ~oreover, they also can be used, in contrast to the above-described compounds, for coupling per se to selectively concentrated substances in foci of disease or certain tissues.
Upon further study of the specification and appended claims, further objects and advantages of this invention will become apparent to those skilled in the art.

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According to the invention, this object is achieved by compounds of general formula I, in which X stands for -O-, -S-, -NR2- with R2 meaning a hydrogen atom or a C16 alkyl radical, Y and Z are the same or different and stand for the radicals -OH, -NHR or -SR with R meaning a hydrogen atom or a C1-6 alkyl radical and U stands for a hydrogen atom, a branched or unbranched C1-6 alkyl, a C16 alkoxy, a hydroxyl or a carboxyl radical, n means the numbers 2 to 6 and m means the numbers 2 or 3 and R is present only if m stands for the number 2, R1 stands for a hydrogen atom, a benzyl radical or a branched or unbranched C06 alkyl radical optionally substituted with one to three hydroxyl, carboxyl or amino groups;
wherein said radicals optionally contain (i) a functional group B, (ii) a compound T which is selectively concentrated in lesions or certain tissues; or (iii) a compound T bound to the radical through a functional group;
and B, i~ Y and/or Z stand for -NHR , stands for an amino, a hydrazino or hydrazide, a carboxyl, a C16-alkynyl or alkenyl, a hydroxyl, an aminophenyl, an oxiranyl, a fluorinated phenoxycarbonyl or a biotin radical;
or, if Y and Z stand for -OH or -SR , B in addition also stands for a halogen, a formyl, a nitrile, a phenylisothiocyanato or a succinimidoxycarbonyl radical ., optionally substituted with a sodium sulfate radical;
and T stands for monoclonal antibodies or their ~ragments, hormones, growth factors, ligands for cell me~brane receptors, steroids, neurotransmitters, fatty acids, saccharides, amino acids and oligopeptides, biotin, as well as radiosensitizers, such as, e.g., misonidazole, and optional]y present funct:ional groups optionally in protected form or their precursors ar~ present in R1, with the exception of compound N{CH2-CH2-CH2-O-C6H3-2,3-(OH)2~3, their technetium and rhenium complexes, as well astheir salts with inorganic and organic acids.
The excluded compound N{CH2-CH2-CH2-O-C~3-2,3-(OH) 2)3 is known (B. Wolff, Angew. Chem. [Appl. Chem.] 98: 173, 1986) and was used for complexing germanium and silicon.
Physiologically compatible inorganic acids, such as, e.g., hydrochloric or sulfuric acid, and organic acids, such as, eOg., acetic or citric acid, are used as acids for salt formation.
A C0 alkyl radical is to be understood to mean a direct bond from the nitrogen atom in Formula I to one of the optional substituents -B, -T or -B-T.
According to the invention, preferred are those compounds in whose general formula I U represents a hydrogen atom and X r~presents an oxygen atom, n is the number 3, Y
2S and Z are the same and stand for OH- or NH2- and R1 represents a hydrogen atom, a benzyl radical, an unbranched C03 alkyl radical optionally substituted with a hydroxyl or amino group, wherein said xadicals optionally contain a functional group B or antibodies or their fragments, steroids or misonidazole bound to the radical through a functional group B.
The substances according to the invention surprisingly have the advantages that they .;: -~4~9 1. can reduce pertechnetate from a nuclide generator under mild conditions, quickly and without adding reducing agents to an oxidation state lower than +7, 2. form stable complexes with the thus reduced techne-tium without further adding reducing agents at neutral pH, 3. are concentrated selectively in certain tissues or lesions because of compounds T, which are coupled with the help of functional group B, such as monoclonal antibodies or their fragments, hormones, growth factors, ligands for cell membrane receptors, steroids, neurotransmitters, fatty acids, saccharides, amino acids and oligopeptides, biotin as well as radiosensitizors, such as, e.g., misonidazole or, without containing a group T, are concentrated in certain tissues or lesions.
The formation of the technetium complexes of the above-described chelates takes place with Tc04- from a nuclide generator with neutral pH without adding reducing agents in aqueous solution.
This property of the chelating agent described here offers significant advantages in comparison with previously known ligands. The incorporation of tin, which has to be added as reducing agent in the known ligand systems, is avoided in the chelate. The formation of the Tc-99m SpeGies, which ~re not bound by the above-described chelates (examples 6 and 7), was not observed. The chelates according to the invention are thus definitely better suited for diagnostic purposes than the previously known chelates.
Their production t~kes place in that amines of general formula II
R --N-~-(CH2)n--Nu)~ (II) in which Nu stands for a nucleofuge, such as, e.g., Cl, Br, I, CH3C6H4S03-, CH3S03- or CF3S03-;

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and R1 stands for a substituent R1, whose optionally present functional groups are present in protected form or as their precursors, and which contains no selectively concentrated compound T, and aromatic substances of general formula III
U' r~ '.
H-X ~ (lil) in which U' stands for a substituent U, whose hydroxy or carboxyl radical is present in protected form, and Y' and Z' stand for Y and Z or their precursors or in protected form, are reacted under base catalysis in polar solvents at temperatures of 50-200 C within 6 hours to 6 days, preferably 2 hours to 4 days, and then functional group B optionally contai~ed in R
or desired aromatic substance substituents Y and Z are -generated, optionally the thus obtained couplable or complexable compounds are coupled with the respective desired selectively concentrating compound T or complexed ~ith the respective desired technetium or rhenium isotope -- and the sequence of the steps coupling on T and complexing with the technetium or rhenium isotope can be interchanged -- and then the still present protective groups are removed or the precursors are converted to the finally desired substituents.
As hydroxy protective groups, e.g., the benzyl, 4-methoxybenzyl, 4-nitrobenzyl, trityl, diphenylmethyl, trimethylsilyl, dimethyl-t-butylsilyl and diphenyl t-butylsilyl groups are suitable. In the case of polyols, the hydroxy groups can also be protected in the form of ketals -`` 20~$9~

with, e.g., acetone, acetaldehyde, cyclohexanone or benzaldehyde. Further, the hydroxy groups also can be present, e.g., as THP ether, ~-alkoxyethyl ether, MEM ether or as esters with aromatic or aliphatic carboxylic acids, such as, e.g., acetie aeid or benzoie acid.
The hydroxy proteetive groups ean be released according to the methods in the literature known to one skilled in the art, e.g., by hydrogenolysis, reductive cleavage with lithium/ammonia, aeid treat~ent of ethers and ketals or alkali treatment of the esters (see, e.g., "Protective Groups in Organic Synthesis," T. W. Greene, John Wiley and Sons 1981).
As acid protective groups, lower alkyl, aryl and aralkyl groups, for example, the methyl, ethyl, propyl, n-butyl, t-butyl, phenyl, benzyl, diphenylmethyl, triphenylmethyl, bis(p-nitrophenyl)-methyl group, as well as trialkylsilyl groups, are suitable.
The cleavage of the protective groups takes place according to processes known to one skilled in the art, for example, by hydrolysis, hydrogenolysis, alkaline saponification of the esters with alkali in aqueous-alcoholic solution at temperatures of 0 to 50C, acidic saponification with mineral acids or in the case of, e.g., tert-butyl esters with the help of trifluoroacetic acid.
Th~ starting materials for the production of the compounds of this invention are all either commercially available or routinely synthesizable by one of ordinary skill in the art using conventional synthetic methods. For the production of compounds of general formula I with Y and Z meaning SH groups, the starting materials are ligand precursors of general formula III with Y' and Z' meaning SR3 radicals. The cleavage of the protective groups after the reaction with the amines of general formula II takes place, 2~8~9 g alternatively with alkali alkylthiolates, alkali alcoholates or alkali metals, preferably with sodium methylthiolate in a polar solvent, preferably in HMPT, DMF or dimethylacetamide.
As amino protective groups, e.g., trifluoroacetyl, t-butoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, benzoxy-carbonyl and acetyl groups are suitable. The amino protective groups can be cleaved according to methods known in the literature, e.g., by basic or acidic hydrolysis, reductive cleavage with zinc in acetic acid or hydrogenolysis.
If the ligand precursor of formula III contains functionalized aromatic substances with Y' and Z' meaning N02 groups, then the production of the chelating agent according to claim 1 takes place by reduction, preferably with tin in hydrochloric acid solution.
Radical R1 can be modified in compounds according to general formula I, in which m means the number 2. If R , for example, is a benzyl group, it can be removed by reaction with hydrogen under increased pressure and increased temperature in the presence of a palladium catalyst. Radical R is then a hydrogen atom.
Rhenium 186 has a physical half-life of 3.7 days and emits beta particles with an energy of 1.1 MeV suitable for the treatment of, e.g., tumors, and, at the same time, gamma radiation with an energy of 137 keV (9% frequency). Rhenium is found in periodic systems in group VII A directly under technetium and exhibits a practically identical structural and chelate chemistry as technetium. These properkies make rhenium 186 into an ideal isotope for therapeutic uses (damage of diseased tissue by particular beta radiation) with the possibility at the same time for diagnostic study of a concentration with the help of the portion of gamma radiation. Rhenium 188, which also can be used for . : .:
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` 2~88~9 treatment of tumors, has a significantly shorter half-life of 17 hours and a beta energy of 2.1 MeV. Also, rhenium 188 has a gamma radiation portion (155 keV; 15%) and can thus also be used for treatment and, at the same time, detection with a gamma camera, e.g., according to the methods disclosed in Fritzburg et al., N. Nucl. Med. 30, 743 (1989).
If the chelating agents complexed with a radioactive isotope show no selectivity for lesions or certain tissues, it is necessary that they be coupled to a selective substance. Radical R is suitable for this purpose, e.g., with the help of functional group B, to produce a stable connection to proteins or other selectively accumulating molecules. By the corresponding selection of the functional group, the coupling is possible under mild reaction conditions, which do not influence the biological function and/or selectivity.
The coupling to the described compounds also takes place according to methods known in the art (e.g., Fritzberg et al.; J. Nucl. Med.: 26, 7 [1987]), for example, by reaction of group B with nucleophilic groups of the selectively accumulating molecule or, if a nucleophile is involved in the case of group B itself, with activated groups of the selectively accumulating molecule.
Group B represents every substituent which, on the one hand, represents a functional group, which makes possible a coupling to a selectively accumulating molecule under mild conditions (e.g., by acylation or amidation) as well as every activated group, which can react with nucleophilic groups of proteins, antibodies, hormones or other biomolecules, such as the amino, phenol, sulfhydryl, aldehyde or imidazole group. By an activated group is to be understood a function which is capable of reacting with the foxmation of a conjugate with a nucleophilic substituent of ~' : . :"' ` ~ ~ ' -,:
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: ' 2 ~ 9 9 a selective molecule or of the complex ligand itself in aqueous solution within a suitably short time, under reac-tion conditions which, result in neither denaturing nor loss of the biological activity or selectivity. Examples in this respect are imide esters, alkylimide esters, amidoalkylimide esters, succinimide esters, acylsuccinimides, phenol esters, substituted phenol esters, tetrafluorophenol esters, anhydrides, hydrazides, alkyl halides and Michael acceptors.
B is preferably a monoanhydride, acid chloride, acid hydrazide, mixed anhydride, activated ester (such as phenol or imide ester), nitrene or isothiocyanate, in particular for the coupling with nucleophilic groups of amino acids or an aliphatic or aromatic primary amine for the coupling to carbohydrate radicals of proteins.
If a nucleophile is involved in the case of group B
itself, it can react with activated groups of a selactively accumulating molecule, and also reacted groups of the selective molecule are enclosed with so-called "crosslinking reagents." These can be homofunctional crosslinkers having two identical functional groups, e.g., imidoester groups or N-hydroxysuccinimide ester (NHS) groups. Alternatively, these crosslinking reagents can be, for example, heterobifunctional "crosslinkers," which contain two different functional groups, for example, two of an NHS
ester, a pyridyl disulfide and an activated halogen, such as an ~-keto-halide. Such crosslinkers can be obtained commercially.
Compounds which selectively accumulate compounds in certain tissues or lesions are used as coupling partners.
Often, the selective accumulating of these suhstances could already be shown by labeling with positron-emitting isotopes (PET technique), iodoisotopes or other coupling partners.
Compounds labeled with Tc-99m have already partially come , ,i'~ ' '.
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into use. See, e.g., Chiton, ~.M. and Witcofski, R.L., Nuclear Pharmacy: An Introduction to the Clinical Application of Radiopharmaceuticals; Lea & Febiger, Philadelphia, PA (1986). But technetium-labeled compounds in such a way have the drawback thAt tin(II) ions have to be added as reducing agent, which results in the above-described consequencss (reduced specific radioactivity and the possibility that unbound Tc-99m together with tin as colloid results in an undesirable storage of radioactivity in other organs and a reduced diagnostic informative value).
Ligands which bind to specific receptors can recognize a changed tissue in their receptor density; they include, i.a., peptide and steroid hormones, growth factors and neurotransmitters. With ligands for steroid hormone receptors, the possibility of an improved diagnosis of breast and prostate cancers was demonstrated (S. J. Brandes & J. A. Katzenellenbogen, Nucl. Med. Biol. 15:53, 1988).
Often, ligands labeled with positron-emitting isotopes could be used for neuroreceptors for the diagnosis of various brain diseases (J. J. Forst, Trends in Pharmacol. Sci. 7:
490, 1987). Occasionally, tumor cells exhibit a changed density of receptors for peptide hormones or growth factors, such as, e.g., the "epidermal growth factor" (EGF). The concentration differences could be used for selective concentration of cytostatic agents in tumor cells (E. Aboud-Pirak et al., Proc. Natl. Acad. Sci. USA 86: 3778, 1989).
Other biomolecules are metabolites that can be put in the metabolism of cells which make a changed metabolism recognizable; they include, for example, lipids (also in the form of liposomes), saccharides, porphyrins, peptides and amino acids. Fatty acids coupled with Tc chelating agents were described in EPA 0 200 492. Other metabolic products such as saccharides (deoxyglucose), lactate, pyruvate and .

:
, ~8~9 amino acids (leucine, methylmethionine, glycine) were used with the help of the PET technique for graphic display of changed metabolic processes (R. Weinreich/ Swiss Med. 8, 10, 1986). Certain porphyrins showed a concentration in tumors (P. A. Scourides, Cancer Res. 47: 3439, l9B7).
Also, nonbiological substances such as misonidazole and its derivatives, which are bound irreversibly to cell components in tissues or tissue parts with reduced oxygen concentration, can be used for specific concentration of radioactive isotopes and thus graphic display of tumors or ischemic regions (M. E. Shelton, J. Nucl. Med. 30: 351, 198g). Other suitable nonbiological substances include cytostatic agents, such as bleomycin, which accumulate in tumors. Also, suitable polymers such as dextrans, polyethylenimines, polyamides, polyureas, polyethers and polythioureas are suitable as coupling partners.
The compounds according to the invention containing biotin make possible the binding of radioactive conjugates to substances containing avidin or streptavidin. This can be used to concentrate antibody-streptavidin conjugates on the tumor and only later to apply the radioactive component containing biotin, which results in a reduced exposure of the patient to radiation ~D. J. Hnatowich et al., J. Nucl.
Med. 28: 1294, 1987). Finally, the direct coupling of the bifunctional chelating agents to proteins, such as, e.g., monoclonal antibodies or their fragments/ albumin, enzymes (e.g., urokinase, streptokinase), fibrin, fibrinogen or myosin, is also possible.
By complexing the conjugates with Tc-99m or rhenium iso~opes, a diagnosis and treatment of tumors or other diseases is made possible. In this case, it is unimportant whether a labeling of the chelating agents with Tc-99m or a rhenium isotope is performed before or after the coupling to ., . :

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the selectively accumulating molecule. But for a coupling to the selectively accumulating molecule after a complexing, the re~uirement is that the reaction of the radioactive complex with the accumulating compound occurs quickly and almost quantitatively under mild conditions, and that no subsequent purification is necessary.
The production of the pharmaceutical agents according to the invention takes place in a way known in the art, in which the complexing agents according to the invention are dissolved --optionally by adding the additives usual in galenicals -- in aqueous medium and then sterilized by filtration. Suitable additives are, for example, physio-logically harmless buffers (e.g., tromethamine~, small additions of electrolytes (e.g., sodium chloride), lS stabilizers (e.g., gluconate or phosphonate) and small amounts of oxidizing or reducing agents (10-500 micrograms/dose). The pharmaceutical agent according to the invention is present in the form of a solution or in freeze-dried form and is mixed shortly before the administration with a Tc-99m-pertechnetate solution, eluted from commercially obtainable generators, or a perrhenate solution.
In the case of the nuclear medicinal in vivo use, the agents according to the invention are administered in amounts of 1 10 to 5 104 nmol/kg of body weight, preferably in amounts between 1 103 and 5 102 nmol/kg of body weight. Starting from an average body weight of 70 kg, the amount of radioactivity for diagnostic uses is between 0.05 and 50 mCi, preferably 5 to 30 mCi per administration.
For therapeutic uses, between 5 and 500 mCi, preferably 10-350 mCi, is administered. The administration is normally performed by intravenous, intraarterial, peritoneal or intratumoral injection of 0.1 to 2 ml of a solution of the .

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agents according to the invention. The intravenous administration is preferred.
The following examples are used to explain the object of the invention in more detail.
Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.
In the foregoing and in the following ~xamples, all temperatures are set forth uncorrected in degrees Celsius and unless otherwise indicated, all parts and percentages are by weight.
The entire disclosures of all applications, patents and publications, cited above and below, and of corresponding application Federal Republic of German P 40 25 788.6, filed August 10, 1990, are hereby incorporated by reference.

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_xample 1:
Tri(B-Carbetho~yethyl)iamine, 1 and di~-carbethoxyethyl)amin~, 2 300 ml of freshly distilled ethyl acrylate is brought to reaction with 300 ml of liquid ammonia for one day in a sealing tube and, after removal of the final ammonia residues from the water bath, the resulting product mixture is fractionated in a vacuum. The first fraction forms bis-(B-carbethoxyethyl)amine, 2, (boiling point 97-110C/0.05 mbar), the main fraction consists of the desired product, 1, (boiling point 120-133 C/0.05 mbar).
Yield: 144 g (4996) of tri(B-carhethoxyethyl)amine, H--NMR (CDCl3, ~, ppm):
4.10 (q, 2H, C(O)OCHzCH3); 2.74 (t, 2H, NCH2CH2); 2.41 (t, 2H, CH2CH2C(O)O);
1.23 (t, 3H, OCH2CH3) 3C-NMR ( CDCl3, ~ , ppm):
172.2 (CH2C(O)O); 60.1 (C(O)OCH2CH3); 49.1 (NCH2CH3);
32.8 (CH2CH2C(O)O); 14.1 (OCHzCH3) For di(B-carbethoxyethyl)amine 2, it was found:
H~ (CDC13, ~ , ppm):
4.12 (q, 4H, C(O)OCH2CH3); 2.88 (t, 4H, NC~2CH2); 2.47 - (t, 4H, CH2C~I2C(O)O); 1.60 (s, br, lH, HN(CH2)2); 1.24 (t, 6H, OCH2CH3) Tris~3-Hydroxypropyl)~mlne, 3 18 g (0.7 mol) of lithium aluminum hydride in 900 ml of absolute ether is suspended in a 2~1iter three-necked flask with dropping funnel and reflux condenser. A solution of 77 g (0.24 mol) Oe ester l in 200 ml of absolute ether is instilled in it within one hour so that the solution boils moderately. After ~ive hours of stirring at 25 C and ., .. . .

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2 ~ 9 careful hydrolysis of excess hydride with water, the product is separated by a Buchner funnel from precipitated hydroxides. After removal of the solvent in a vacuum, the residue is briefly boiled up in ethanol, additional LiAl(OH)4 is filtered off by suction by a îrit (D3), the alcohol is drawn off and the remaining liquid is taken up in methylene chloride. Pressureless filtering yields a honey yellow, highly viscous liquid by another frit (D4) and removal of the solvent in a vacuum.
Yield: 27.3 g (59%) H-N~DR (CDC13, ~, ppm) 3.78 (t, 2H, CH2CH2OH); 2.62 (t, 2H, NCH2CH2); 1.80 (q, 2H~ CHzCH2CH2);
C-N~rR (CDC13, ~, ppm):
1560.6 (CH2CH2OH); 51.3 (NCH2CH2); 2~-1 (C~IzCH2CH2) Tri~(3-Chloropropyl)amine, 4 18.9 g (160 mmol) of thionyl chloride is added to 8.6 g (45 mmol of tris(3-hydroxypropyl)amine, 3, dissolved in 80 ml of chloroform. Thus, an insoluble white mass results, which slowly dissolves again. After the reaction mixture has been refluxed for three hours, excess SOCl2 is hydrolyzed with water after cooling off. The organic phase is shaken out four times wi~h 50 ml of hot water, the combined aqueous phases are made strongly alkaline with 40%
sodium hydroxide solution and then are extracted four times with 80 ml of ether each. After drying on Na2SO4 and removal of the ether in a vacuum, the yellowish residue is fractionated, and the product goes over as colorless liquid and crystallizes out after prolonged standing at room temperature. Tris~3-chloropropyl)amine can be recrystalli~ed from ethanol (3 g of 4 for 7 ml of ethanol).

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2~ 8~9 Yield 9.6 g (87~) ~elting point: 35C
Boiling point: 120 C/0.05 mbar H-NMR (CDC13, ~, ppm):
3.60 ~t, 2H, CHzCH2Cl); 2.52 (-t, 2H, NCH2CH2); 1.88 ppm (q, 2H, CHZCH2CH2) C-NMR (CDCl3, ~, ppm):
50.5 (CH2CH2Cl); 43.0 (NCH2CHz); 30.1 (CH2C~2CH2) 2,2-Dimethyl-1,3-benæodioxol-4-ol, 5 77 g (0.61 mol) of pyrogallol in 250 ml of abso]ute toluene is suspended and heated in a 500-ml two-necked flask with dropping funnel and a Widmer spiral column. When the solvent begins to boil, 75 ml (0.61 mol) of 2,2-dimethoxypropane is added. After that, distillate lS continuously passes over at the column head at about 60C.
After t~o hours, another 75 ml of dimethoxypropane is added.
After the temperature at the column head drops ~about six hours), the reaction mixture is refluxed overnight for completion of the reaction. The cooled solution is freed in a vacuum from toluene and the viscous residue is distilled over a bridge with large cross section. As a result, the product already crystallizes out on the bridge wall and has to be transferred to the receiver by heating. The distillation temperature is selected so that existing yellowish impurities distill over only to a limited extent.
The slightly yellowish product can be sublimated at 75 C, 0.2 mbar. The acetal is readily soluble in acetone and methanol, slightly soluble in chloroform.
Variants for working up: After removal of the toluene, the crude product is taken up in so much hot carbon tetrachloride, that it just about dissolves. Wlth cooling off to room temperature, white product 5 crystallizes out '20~99 from the solvent. It is sublimated according to the same conditions as described. A significant difference in the yields is not observed.
Yieldi 50 g (50%) Melting point: 90C
H-NMR (CDCl3, ~, ppm):
6.68 (t, lM, Ar-~); 6.46 (d, lH, Ar-H); 6.40 (d, lH, Ar-H); 5.13 (s, lH, Ar-OH); 1.69 (s, 6H, CH3) C-NMR (MeOD, ~, ppm):
149.7, 141.8, 135.3, 122.1 (Ar); 118.6 (C(CH3)2);
111.3 (Ar); 25.8 (C(CH3)2) Tris~3-(2,2-Dimethyl-1,3-benzodio~ol-4-ylo~y1-propyl~-amine, 6 20.1 g (121 mmol) of dry pyrogallol acetal 5 is intro-duced in a stung-out 250-ml two-necked Schlenk flask with a reflux condenser and dissolved in 100 ml of absolute (99%) ethanol. For removal of the final oxygen residues, the flask is alternately degassed five times and aerated with argon. Then, 4.75 g (121 mmol) of potassium ~etal is added in small pieces. First, white potassium phenolate preci-pitates from the solution, which is at once dissolved again.
Now, a li~ewise degassed solution of 9.6 g (39 mmol) of tri-(3-chloropropyl)amine and 20 ml of ethanol is sprayed in the now extremely oxygen-sensitive solution. After the solution has been xefluxed for four days, 4 ml of glacial acetic acid is added, precipitated KCl is filtered off from the still hot but no longer air-sensitive solution and the filter cake is rewashed with a little hot ethanol. By allowing it to stand at room temperature, the ligand precursor crystallizes out slowly. The crystals are filtered off and rewashed with ice-cold ethanol. A second fraction can be obtained by concentrating the mother liquor by evaporation.

- ; .. . - , ,. , : : , ~:
- : , . :: . .

2 ~

Yield: 11.5 g (46%) Melting point: 65 C
H-NMR (CDCl3, ~, ppm):
6.66 ~t, lH, Ar-H); 6.42 (cl, lH, Ar-H); 6.26 (d, lH, Ar-H); 3.95 (t, 2H, CHzCH2O); 2.59 (t, 2H, NCHzCH2);
1.91 (q~ 2H, CH2CH2CH2); 1.69 (s, 6H, C(CH3)2) 3C-NMR (CDCl3, ~, ppm):
148.3 (Ar); 143.2 (Ar); 135.2 ~Ar); 121. (Ar); 117.9 ~C(CH3)2); 107.9 (Ar); 102.0 (Ar); 66.8 (CHzCH2O); 49.8 (NCH2CH2); 26-9 (CH2CH2CH2); 25.7 (C(CH3)2 Tris-~3-~2,3-Dihydroxyphenoxy)propyl)aminohydrochloride, 7 39.6 g (62 mmol) 6 is dissolved under argon in 250 ml of glacial acetic acid and heated to boiling. 200 ml of a mixture of 50% glacial acetic acid, 20% water and 30%
fuming hydrochloric acid is instilled in it within two hours. The solvent is distilled of~, iso that about 200 ml remains in the flask. The solution is slowly cooled off. The precipitated yellowish crystals are filtered off and recrystallized in a little hot glacial acetic acid. The white powder thus obtained is dried at 90 C, 10 mbar for two days on an oil pump vacuum.
Yield: 31.7 g (92%) Melting point: 190C
H-NMR([D6]-DMSO, ~, ppm):
10.47 (s, br, 1/3H, NH); 8~93 ~s, br, lH, OH); 8.19 (s, br, lH, OH); 6.42-6.54 (m, 3H, Ar-H);
4.02 ~m, br, undissolved, 2H, CH2CH2O); 3.40 (m, br, undissolved, 2H, NCH2CH2);
2.17 (m, br, undissolved, 2H, CH2CH2CH2) 13C NMR (tD6]-DMSO, ~, ppm):
147.5, 146.2, 134.7, 118.5, 109.6, 105.2 (Ar); 66.2 (CH2CH2O); 49.9 (NCH2CH2); 23.4 (CH2CH2CH2) ~4~X!~3 Example 2:
Benzyl ~ aarbeth~yethyl)amine, ~
172 g (1.60 mol) of benzylamine is introduced in 500 ml of ethanol and mixed under ice cooling with 384 g ~3.84) mol of acrylate. The reaction mixture is stirred for 5 days at room temperature. Solvents and excess feedstocks are drawn off in a rotary evaporator. The remaining solution is fractionated in a vacuum.
Fraction 1: less than 140 degrees/0.05 mbar Fraction 20 140-145 degrees/0.05 mbar Fraction 3: 145-148 degrees/0.05 mbar Fxaction 4: 145-150 degrees/0.05 mbar Yield: 380 g (77%~ 8 from fraction 4 1~-NMR (CDCl3, ~, ppm):
7.27 (m, 5H, Ar-H); 4.10 (q, kH, C(O)OCH2CH3); 3.59 (s, -2H, C6HsCH2N); 2.80 (t, 4H, NCH2CH2); 2.46 (t, 4H, C~2CH2C(O)); 1-23 (t, 6H, OC~zCH
C-NMR (~D30D, ~, ppm):
173.7 (c~2c(O)O); 140.2, 129.7, 129.0, 127.9 (ar); 61.1 (OC~2CH3); 59-1 (C6HsC~ZN); 51.1 (NCH2CH2); 33.5 (CH2CH2C(O)); 14-5 (OCH2CH3) Benzyl-bis(3-hydroxypropyl)ami~e, 9 1~ g (0~5 mol) of lithium aluminum hydride is introduced in 900 ml of ether and 92 g (0.3 mol) of ester 8 is instilled slowly under ice cooling. The solution is stirred for 12 hours at room temperature and then carefully hydrolyzed with water. The ether and the aqueous phase are decanted from precipitated LiAl(O~) 4 . The solid is washed several times with ether. The combined organic phases (ether and ethanol) are separated from water in a separating unnel, dried on MgSO4 and filtered. After removal of the , 8 ~ 3 solvent in a vacuum, the product: remains as colorless liquid.
Yield: 60.5 g (90%) l~-NMR (CdCl3, ~, ppm~:
7.30 (m, 5H, Ar-H); 4.11 (s, br, 2H, C~2OH); 3.67 (t, 4H, CH2CH2OH); 3.56 (s, 2H, C6HsCH2N); 2.61 (t, 4H, NCH2CHz); 1-75 (q, 4H, CH2CH2CH2 C-NMR (CD30D, ~, ppm):
137.8, 130.1, 129.2, 128.0 (Ar); 61.9 (CH2CH2O); 59.5 (C6H5CH2N); 52.3 (NCH2CH2); 30-2 (CH2CH2CH2) Benzyl-bi~3-chloropropyl)amine, 10 143 g (0.64 mol) of 9 is introduced in 600 ml of chloroform. 182 g (1.53 mol) of thionyl chloride, dissolved in 100 ml of chloroform, is instilled in it at room 15 temperature. The addition has to take place so that the solvent boils moderately. After completion of the addition, it is refluxed for 3 hours. The cooled solution is carefully hydrolyzed with water and washed t~,rice with 300 ml of hot water. Now, the organic phase is strongly 20 concentrated by evaporation and further shaken out twice with 250 ml of hot water each. After combining the aqueous phases, the latter are made strongly alkallne with sodium hydroxide solution (40%) and extracted twice with 400 ml of ether each. The combined ether extracts are dried on sodium 2S sulfate. Then the solvent is drawn off on a rotary evaporator. The crude product is fractionated.
Yield: 140 g (84%) Boiling point: 114-125 C
1H-NMR (CDCl3, ~, ppm):
7.30 (m, 5H, Ar-H); 3.58 (t, 4H, CHzCHzCl); 3.55 (s, 2H, C6HsCH2N); 2.S~ (t, 4H, NCHzC~2); 1.92 (q, 4H, CH2CH2CH2 ) :, . .

: . :~

.:
~ ,.. . .

" ~0'~8~9 C-NMR (CD30D, ~, ppm):
140.3, 129.9, 129.2, 128.0 (Ar); 59~8 (C6HSCH2N); 51.9 (NCH2CH2~; 43.8 (CH2CH2Cl); 31.3 (C~I2C~I2CH2) senzyl-bi~3-~2,2-dimethyl-1,3-benzodioxol-4-yloxy]propyl~amine, 11 13.3 g (80 mmol) of ketal 5 is dried for a half hour in a high vacuum at 50 degrees in a stung-out 250 ml two-necked Schlenk flask. Then 5 is dissolved in 100 ml of absolute (99%) ethanol. The solution is evacuated several times until boiling and aerated with argon~ Potassium (3.1 g, 80 mmol), which is cut until clear, is added and oxygen-free chloride 10 (9.7 g, 37.3 mmol) is added to the now oxygen-sensitive solution. The batch is refluxed for 3 days. 3 ml of glacial acetic acid is added for working up and the precipitated potassium chloride is filtered off still hot.
The filtrate is drawn dry and dissolved in a mixture of about 30 ml of pentane/ether 1:1. The brown solution is eluted on a short column (about 50 g of silica gel) with ether/pentane 1:1. Here, attention must be paid that dark-colored products are not coeluted. Aft~r the removal of the mobile solvent, a yellow oil remains, which is recrystallized from ethanol (16 g in 200 ml of ethanol). In doing so, the product accumulates at room temperature as colorless crystals.
Yield: 16 g (80%) Melting point: 46-47C ;;
H-NMR ( [ D6] -acetone, ~, ppm):
7. 30 (dd, 2H, benzyl aromatic substance); 7 . 20 (m, 3H, benzyl aromatic substance); 6.67 (dd, 2H, catechol);
6. 41 (dd, 4H, catechol); 4 . 07 (t, 4H, CHzCH20);
3 . 58 (s, 2H, C6HsCH2N); 2 . 61 (t, 4H, NCH2CEI2);
1.91 (q, 4H, CH2CH2CH2); 1.60 (s, 12H, C(CH3)2) - . .
.

C-NMR (CD30D, S, ppm):
149.3, 144.1 (catechol); 140.7 (benzyl aromatic substance); 136.1 (catechol~; 129.4, 128.8, 127. 4 (benzyl aromatic su~stance); 121.9 (catechol); 118.4 (C(CH3)2); 109.4, 102.6 (catechol); 67.8 (CH2CH2O); 59.3 (C6H5CH2N); 50 7 (NCH2C~I2); 27.9 (CHzC~2CH2); 25.8 (C (CH3) 2) Benzyl-bis t ~3- ~2,3-dihydroxyphenoxy)propyl]aminohydrochloride, 12 14 g (27 mmol) of ligand precursor 11 is dissolved in 100 ml of glacial acetic acid and heated to boiling. 100 ml of an acid mixture (50% glacial acetic acid, 20% water, 30%
fuming hydrochloric acid) is added to it within two hours, and the liquid loss resulting from distilled-off solvent is compensated for by acetone that is being liberated. A~ter completion of the addition, so much solvent is distilled off that about 50 ml remains in the flask. The hot solution is slowly cooled off. But the product cannot be precipitated in this way. If all solvent is removed, the ligand accumulates as voluminous residue. This crude product is liberated from the final acetic acid residues by washing with ether and is dried in a high vacuum.
Yield: 12.4 g (96%) H-NMR( [D6]-DMSO, ~, ppm):
10.85 (s, br, lH, NH); 8.98 (s, 2H, OH); 8.17 (s, 2H, OH); 7.65 (d, 2H, benzyl aromatic compound); 7.43 (m, 3H, benzyl aromatic compound); 6.53 (t, 2H, catechol);
6.43-6.38 (m, 4H, catechol); 4.39 (s, br, 2H, C6HsCH2N);
~ .97 (t, br, CH2CH2O); 3.27 (t, br, NCH2CH2); 1 92 (q, br, 4M, CH2CH2CH2) . ,. . . . - . : ~.;: .

'', ' ~ .' ' , : ' ,:
': ' . , ' ~ ~ i ' , ' ' ' , ' ;; ' ~

2 Q ~ 9 C-NMR (CD30D, ~, ppm):
148.3, 147.0, 135.7 (catechol~; 132.2, 131.1, 130.4, 130.4 (benzyl aromatic compound): 120.3, 110.6, 106.6 (catechol); 67. 9 (CH2CH20); 58. 5 (C6HsCH2N); 52.5 (NCH2CH2); 24~ (CH2CH2CH2) bis~3-~2,3-Dihydro~yph~noxy)propyl)ami~ohydrochloride, 13 10 g (21 mmol) of 17 ~ is dissolved in 300 ml of absolute methanol and mixed with 2 g of Pd (OH) 2/C (20~). In a hydrogenation unit, the mixture was shaken with a hydrogen pressure of 3 bars for six hours at room temperature. The catalyst is filtered off and the solvent is drawn off. The oily residue is dried ~or 24 hours at 50C in a high vacuum.
Yield: 7 g (70%) H-NMR ([Ds~-pyridine, ~, ppm): ~
8.96 (s, 6~, OH and NH2); 6.90-6.30 (m, 6H, catechol); ::
3.97 (t, ~H~ CH2C~20); 3.17 (t, 4H~ NCH2CHz); 2-2~ (q~ ~`
4 H r CH2cH2cH2 ) C-NMR (~Ds]-pyridine, ~, ppm):
148.8~ 148.2~ 136.5, 119.7, 111.0, 106.0 (catechol);
67.6 (CH2CHzO); 46. 5 (NCH2CH2); 26. 8 (CH2C~2CH2) , ~ .
Example 3:
bis~3-(2,2-Dimethyl-1,3-be~zodio~ol-4-~loxy)propyllamine, 14 17.9 g (34.5 mmol) of 11 is dissolved in a hydrogenation flasX in 300 ml of absolute methanol, mixed with 2.0 g (2.8 mmol) of catalyst (Pd(OH)2/C) and shaken for four hours in a hydrogen hydrogenation apparatus at 3 bars ~:
of H2 pressure and 25 C. Then, the catalyst is filtered off and the solvent as well as resulting toluene are removed in a water j`et vacuum. The resulting oil is dried at 60C/0.05 mbar for six hours.

,, , , . : .
, '. ' , ~ ' ; .

2~ 9 Yield: 12.9 g (87%) H-N~DR(CDCl3. ~, ppm):
6.69 (dd, 2H, cat2chol); 6.46 (d, 2H, catechol); 6~43 (d, GH, catechol); 4.14 (t, 4H, CE~zCH20); 2. 83 (t, 4H, NCH2CH2); 1.99 (q, 4H, CH2CH2CH2); 1.68 (s, 12H, C(CH3)2) 3_NMR([D6] -benzene, ~, ppm):
149.3, 143.9, 136.2, 121.6 (catechol); 117.9 (C(CH3)2);
109.1, 102. 6 (catechol); 6~.9 (CH2CH20); 46.6 (NCH2CH2);
29. 8 (CH2CH2CH2); 2 5. 7 (C( CH3)z bis~3-(2,3-Dihydroxyphenoxy)propyl)aminohydrochloride, 13 8 g (18.6 mmol) of 14 is dissolved in 80 ml of glacial acetic acid and mixed in boiling heat within two hours with 80 ml of an acid mixture (50% glacial acetic acid, 30%
water, 20% fuming hydrochloric acid). In this case, the solvent distills off with acetone that is liberated as azeotrope. After completion of the addition, it is distilled for another half hour. Then, the residual solvent is drawn off on a rotary evaporator. The remaining residue is dried at 60C/O.OS mbar for six hours in a high vacuum.
The powdery product is washed with ether. Ether residues are then removed on the oil pump.
Yield: 6.25 g (87~) The analytical data is identical with that of 13 from 12.

Example 4:
2,3-Dinitrophenol, 15 15.0 g (108 mmol) of 3-nitrophenol is dissolved in 150 ml of ethanol. 30 g (124.5 mmol) of Cu(N03)2-~3 H20) is added to it. The reaction mixture is then refluxed to boiling for 20 hours. The solvent is drawn off at a rotary evaporator. The solid residue is dissolved in 2 M HCl and extracted four times with 50 ml of ether each. The combined ether extracts are dried on Na2SO4 and liberated from solvent. The orange solid (21 g) is chromatographed on a short column on about 60 g of silica gel with petroleum ether/ethyl~ First, 3,6-dinitrophenol is eluted, followed ~y 3-nitrophenol and 3,4-dinitrophenol. Finally, the desired 2,3-dinitrophenol is obtained. The desired product can be recrystallized from benzene/petroleum ether (7:93, v:v) .
Yield: 2.9 g (14.6%) Melting point: 146C
H-NMR (CDCl3, ~, ppm):
7.18-7.79 (m, 3H, ArH); 9.90 (s, br, lH, Ar-OH) 13C-NMR (CD30D~ ~ ppm):
151.87 (C-OH); 142.17 (C-NO7(m); 134.22 (C-NO2(o));
132.27 (C-H0; 124.54 (C-H); 116.12 (C-H) Be~zyl-bis L (3-(2,3-dinitrophenoxy)propyl]amine, 16 0.92 g (5 mmol) of 2,3-dinitrophenol is dissolved in 10 ml of ethanol. 0.28 g (5 mmol) of KOH in 25 ml of ethanol is added to it under argon. In doing so, the potassium salt precipitates as red solid. 0.65 g (2.5 mmol) of benzyl-bis(3-chloropropyl)amine, 10 in 5 ml of ethanol is added to it, and the solid is partially dissolved. With subsequent heating to the boiling temperature of the ethanol, the solid is completely dissolved. The red solution is now refluxed for 24 hours. A precipitate of KCl forms. The suspension is filtered hot. With cooling, 16 crystallizes out in the form of colorless feathers~ The product is recrystallized from ethanol.
Yield: 0.76 g (55-~) Melting point: 98-104C

~ :
..

g~'3 - 2~ -H-NMR(cDcl3~ ~, ppm)O
1.91 (q, 4H, CH2CH2CH2); 2.60 (t, 4H, NCH2); 3.58 (s, 2H, C6HsCH2); 4.15 (t, 4H, OCH2); 7.21 (s, 5H, benzyl-H); 7.22-7.81 (m, 6H, catechol-H) C-NMR (CDCl3, ~, ppm):
26.70 (CH2CH2CH2); 49.81 (NCHz); 58.77 (C6HsCH2); 68.57 (OCH2); 116.08, 119.56 (CH(phenyl)): 126.89, 128.20, 128.74 (CH(benzyl)): 131.04 (CH(phenyl)): 134.99 tC(NO2-o)): 139.06 (C(benzyl)): 140.59 (C(NO2~m)), 151.37 (C(O-phenyl)) Benzyl-bi~C3-(2,3-diaminophenoxy)propyl]amine, 17 1.06 g of tin (8.93 ~mol) is added to 5 ml of concentrated hydrochloric acid. A solution of 0.5 g (o.9 mmol) of 16 in 5 ml of methanol is sprayed to it. The reaction mixture is heated for 30 minutes to 50C, and its color becomes brown. Then the reaction mixture is poured into a solution of 2.5 g of NaOH in 50 ml of water. This mixture is extracted five times with 15 ml of ether each.
The ether extracts are washed with water and dried on Na2SO4. After the removal of the ether, a brownish oil remains, which is not further purified.
Yield: 0.275 g (70%) H-NMR (CDCl3, ~, ppm):
1-99 (q, 4H, CH2CH2CH2); 2.69 (t, 4H, NCH2); 3.32 (s, 8H, NH2); 3.65 (s, 2H, benz-CH2); 4.04 (t, 4H, OCH2);
6.28 6.80 (m, 6H, phenol-H); 7.31 (s, 5H, benz-H) 3C-NMR (CD3Cl, ~, ppm) 27.14 (CH2CH2CHz); 50.24 (NCH2); 58.69 (C6HsCH2); 62.23 (OCH2); 103.38, 109.50, 118.99 (CH(phenol); 123.67 (C(NH2-o)); 126.69, 128.06, 128.59 (CH(benzyl)); 135.32 (C(NH2~m)); 139.59 (C(ben2yl)); 147.71 (C(O-phanol) .

,.: :::,.'~ ' .:.,.:
~. . 1.

~ O ~ 9 bi~[3-(2,3-Diaminophenoxy)propyl~amine, 18 846 mg (1.66 mmol) of ligand 17 is dissolved in 50 ml of methanol. 0.09 g of Pd(OH)2 (on carbon, 10%) and 5 ml of hydrazine hydrate (80% in water) are added to it. The reaction mixture is heated to boiling for 10 hours. The resulting suspension is filtered and evaporated to dryness.
The product is soluble in methanol and, after adding diethyl ether, precipitates as light greenish oil. This oil is liberated from solvent residues on the oil pump, and a gray powder is obtained.
Yield: 400 mg (69%) H-NMR (CD30D, ~, ppm):
1.99 (q, 4H, CH2CH2CH2); 2.89 (t, 4H, NCH2); 4.05 (t, 4H, OCHz); 6.24-6.68 (m, 6H, Ar-H) C--NMR (CD30D, ~ , ppm):
29.77 (CH2CH2CH2); 47.06 (NCH2); 67.70 (OCHz); 104.39, 111.13, 120~08 (CH(phenol)); 124.47 (C(NH2-o) ); 136.70 tC(NHz~m))~ 148.87 (C(O-phenol)) Example 5:
tris[3-(2,3-Dinitrophenoxy~propyl]amine, 19 1 g of 2,3-dinitrophenol 15 (5.43 mmol) is stirred under reflux with 0.446 g of tris(3-chloropropyl)amine, 4 (1.81 mmol) and 0.305 g of KOH (5.43 mmol) in 50 ml of ethanol for 10 hours. Then, the reaction mixture is allowed to cool off and the precipitated solid is filtered off. The solid is taken up in acetone. In this case, organic components are dissolved while the formed KCl remains.
Insoluble components are separated by filtration. The acetone solution is concentrated by evaporation to 20 ml and cooled to 4C. After 24 hours, 19 can be isolated in the form of colorless needles.

.

, '' '.

~' . `, . . .

~4~9 Yield: 638 mg ~51%) Melting point: 145-147C
H-NMR(~D6]-acetone, ~, ppm):
1.93 (q, 6H, C~2CH2CH2); 2.59 (t, 6H, NCHz); 4.31 (t, 6H, OCH2); 7.63-7.88 (m, 9H, Ar-H) trist3 (2,3-Diaminophenoxy)propyl]amine, 20 638 mg of 19 t0.925 mmol) is added to a mixture of 5 ml of HCl (conc.), 5 ml of methanol and 1.64 g (13.8 mmol) of tin. The reaction mixture is stirred under reflux for 2 hours. The reaction is completed when the tin has completely dissolved. After the cooling, the green reaction solution is made strongly alkaline (pH 13) with an excess of KOH and shaken out twice with 15 ml of acetone each. The combined acetone extracts are mixed with 5 ml of water and shaken out twice with 30 ml of ether each. The ether extract is dried on sodium sulfate. The solvent is removed in a pump vacuum. A yellowish oil is obtained which solidifies with drying in a pump vacuum.
Yield: 638 mg ~51%) 1H-NMR (CDCl3, ~, ppm):
1.92 (q, 6H, CH2CH2CHz); 2.65 (t, 6H, NC~z); 3.36 (s, 12H, NH2); 3.97 (t, 6H, OCH2); 6.23-6.74 (m, 9H, Ar-H) Example 6:
Productio~ of the technetium complexe~ o~ 12 and 13 Methanol solutions of ligands 12 and 13 (30 mmol/l) are produced. 8 microliters of such a solution is mixed with 10-40 microliters of a saline solution made from a Tc/ Mo reactor. The resulting solution is immediately examined by thin-layer chromatography. This examination (mobile solvent THF) shows a complete incorporation of 9mTc in the ligands.
The Rf values are 0.3 for Tc04, 0.65 for Tc 12 and 0.60 ., . . :,, .: . . ., -: :, ' ',, ; , ~ , :
. ,:.:. :.. : .

for ~c 13. Residues of pertechnetate can be easily discovered in these various Rf ~alues, but are not ~ound.

Example 7:
Productio~ of the tech~etium complex of 17 A methanol solution of 17 with the concentration of 50 mmol/l is produced. One microliter of this solution is mixed with 50 microliters of an eluate solution of a Tc/ Mo generator. lO microliters of a 0.1 N NaOH and 10 microliters of phosphate buffer (ionic strength of 0.1, pH
lo 7) are added to it. The reaction mixture is allowed to stand for 5 minutes at room temperature and then is characterized by thin-layer chromatography (mobile solvent THF). This analysis shows the complete incorporation of ~ c in the ligands. The Rf values are 0.15 for ~ Tc04 and 0.24 for Tc 17.

Example 8:
siotin ~HS
1.72 g ~8.19 mmol) of DCC is added to a solution of 2.0 g (8.19 mmol) of D(+)biotin and 1.23 g (10.66 mmol) of N-hydroxysuccinimide in 25 ml of DMF. The resultingsuspension is stirred for 24 hours at room temperature. The solid is filtered off and the solution is cooled for 4 hours to -16 C. It is filtered off from the precipitated solid and the solvent of the filtrate is drawn off in a vacuum.
The colorless residue is wash~d several times with ether and finally dried.
Yield: 2.47 g (89~) -" 2~8.~9 H-NMR([D6]-DMSO, ~, ppm 400 MHz):
6.4~, 6.37 (s, br, 2H, b and g); 4.29 (m, br, 1~, c);
4.13 (m, br, lH, f); 3.09 (m, br, lH, e); 2.82 (dd, J=12 Hz, 6 Hz, lh, d); 2.80 (s, 4H, n); 2.66 (t, J=7 Hz, 2H, k); 2.57 (d, J=13 Hz, lH, d); 1.63-1.50 (m, 6H, h~
C-NMR ([D6]-DMS0, ~, ppm):
170.1 (2C, m); 168.8 (l); 162.6 (a); 60.9 (f), 59.1 (c); 55.1 (e); 39.8 (d); 29.9 (k); 27.7, 27.5 (i and j); 25.4 (2C, n); 24.2 (h) b HN NH~
H~H o S ~ ~o ~ , Diagram for the allocation of the NMR signals for blotin NHS.

Coupling of biotin NHS to bis~3-(2,2-dimethyl-1,3-benzodioxol-4-yloxy)propyl]amine 860 mg (2.0 mmol) of ligand precursor 14 is stirred in 30 ml of degassed DMF with 670 mg (2 mmol) of bioti~ NHS and 860 mg (8 mmol) of triethylamine for 80 hours at room temperature. Then, the reaction mixture is mixed with 80 ml of degassed watex and cooled for 1 hour to 4 C. I'he ~0 precipitated sol:id is isolated by filtration and taken up in 50 ml of acetone. After the removal of the solvent, the .. . ~ . -, . ,, . :
.. i~ . .:

-- 2 0 ~ 9 - 34 ~

Cleavage o tha protuoti~e groups of conjugat~ biotin-14 1 g of the biotin-14 conjugate is dissolved in 20 ml o~
methanol. 4 ml of fuming HCl is added and it is stirred at room temperature for 3 days. The solvents are removed in a S vacuum and the solid residue is dissolved in methanol. The purification takes place chromatographically on SiO2 with methanol/THF (1:1) as mobile solvent. ~he cleavage of the protective groups in the absence of the signals for the acetal unit is detected in the H-NMR spectrum. This observation is also confirmed by the C-NMR spectrum.
Yield: 53%
Because of the problems in the cleavage of the protective group, an alternative method of synthesis was also examined.
:" .
~xample 8a:
Tetratdimethylltert-butyl)]~ilylether of 13 3.85 g (10 mmol) of 13 and 6.55 g (96 mmol) of imidazole in 35 ml of DMF are introduced in a 100 ml Schlenk flask and placed under protective gas. Then, solid TBDMSCI
(7.5 g, 50 mmol) is added, and heating takes place. It is stirred for 20 hours at room temperature. Then, the solution is mixed with 200 ml of ether. This mixture is washed three times with water fremoval of the DMF) and dried on sodium sulfate. After the filtering off of the drying agent, the ether is drawn off on a rotary evaporator. A
~right green liquid remains. From this, the product can be isolated on SiO2 by column chromatography (pentane:ether 1:2~.
Yield: 7 g (86%) .

,, , . ~ "
., , ~ .

2 ~ 9 H--NMR (CDCl3, ~ , ppm):
6.71 (t, lH, catechol); 6 . 50 (d, lH, catechol); 6 . 48 (d, lX, catechol); 3 . 99 (t, 2H, CH2C~2O); 2 . 81 (t, 2H, N(:H2CH2~; 2.01 (q, 2H, CH2CH2CH2); 1.01 and 0.97 (s, 18H, ((CH3)3C-Sio); 0.21 and 0.14 (s, 12H, (CH3)2 sio C-NMR (CDC13, ~ , ppm):
151.9, 148.0, 136.3, 120.2, 113.7, 105.9 (catechol);
66. 8 (CH2CH20); 46 . 9 (NCHzCH2); 30. 0 (CH2CH2CH2); 26 . 1 and 26.0 (6C, (CH3)3CSio); 18 . 6 (2C, (CH3)3CSio); -3 . 8 and -4.0 (4C(CH3)2Sio) MS (70 eV):
806 (2) M ; 749 (23) M -C(CH3)3;
452 ~100) M-C6H3(OH) (osi(cH3)2c(cH3)3)2 Coupling of tetra[dimethyl(tert-butyl]silylether of 13 to biotin 1.64 g (4.84 mmol) of biotin NHS and 3.90 g (4 . 84 mmol) of tetra[dimethyl(tert-butyl)]silylether of 13 are stirred with 2.02 g (20 mmol) of triethylamine in 70 ml of DMF for 3 days under argon. The solvent is drawn off in an oil pump vacuum and the remaining oil is washed several times with water. The thus obtained crude product is chromatographically (sio2~ methanol) purified. The identity - of the product could be shown by nuclear resonance spectroscopy.
Yield: 43%

Cleavage of the protective group~ of conjugate bi~tin- -tetratdimethyl~tert-butyl)]silylether o~ 13 1.03 g (1 mmol) of (T8DMS)-13-biotin conjugate is stirred for 24 hours in a mixture of 2 ml of HCl (conc.~ and 20 ml of THF. Then, it is mixed with 20 ml of water and the THF is removed in a vacuum. The conjugate precipitates in , :; i : : ,, :;- , , : ~ . . ! ; ' ;; ' , .; , ;

2 ~ 9 doing so. The hydrochloric water solution is decanted and the residue is washed several times with water. The residue can be recrystalli~ed from methanol.
Yield: 45%
lH~ CDCl3, ~, ppm) 6.70-6.20 (m, 6H, catechol); 4.25 (m, lH, NH-CH-CH2);
4.15 ~m, lH, CH2-CH-CH); 3.92 (m, undissolved, 4H, O-CH2-CH2); 3.48 (m, undissolved, 4H, N-CH2-CH2); 3.10 (m, lH, CH-CH(R)-S3; 2.75 (d, lH, CH-CH(H)-S); 2.61 (sbr, lH, CH-CH(H)~S); 2.28 (tbr, 2H, NC(O)-CH2-CH2); l.g5 (m, undissolved, 4H, CH2-CH2-CH2-O); l.S5-1.30 (m, 6H, biotin alkyl chain) Example 9~
tert-Butyl ethyl acetate of 14 2.7 g (6.3 mmol) of 14 is dissolved in 40 ml of THF/X20 (9:1) and mixed with 0.67 g (1 eq.) of Na2CO3. 2.46 g (2 eq.) of bromoacetic acid-tert~butyl ester is added and stirred at room temperature for 24 hours. Then, 80 ml of CH2C12 is added and dried on ~qgSO4. After the filtering off of the drying agent, the solution is rigorously concentrated by evaporation. The residue is taken up several times in hexane and decanted. The crude product is purified by column chromatography (70 g of SiO2,~pentane/ether 3:1).
Yield: 3.3 g (95%) 1H-NMR (CDCl3, ~, ppm):
6.90-6.30 (m, 6H, catechol); 4.11 (t, 4H, OCH2CH2);
3.26 (s, 2H, OC(O3CH2N); 2.81 (t, 4H,~ NCH2CH2); 1.93 (q, 4H, CH2CH2CH2); 1.69 ~s, 12H, C(CH3)2); 1.45 (s, 9H, (CH3)3COC(O)) .

~ o ~

Gly--13 3.10 g (5.7 mmol) of the product of the preceding reaction is treated in 50 ml of methanol in boiling heak with 50 ml of acid mixture analogously to the production of 13 from 12. After completion of the reaction, the solvent is drawn off on a rotary evaporator. In this case, the product precipitates so that the aqueous phase can be decanted. The product is dissolved in acetone and liberated from the solvent in a vacuu~. In this case, it accumulates as white powder.
Yield: l.gO g (75~) H-NMR (CDCl3, ~, ppm):
6.90-6.30 (m, 6H, catechol); 4.17 (t, 4H, O-CH2~CHz);
3.80 (t, 4H, NCH2CH2); 3.60 (s, 2H, HOOCCH2N); 2.5 (q, 4H, C~2CH2C~2) Pr~duction o~ the NHS e~ter of gly-13 0.464 g (2.25 mmol) of DCC is added to a solution of 1 g (2.5 mmol) of gly-13 and 0.388 g (2.4 mmol) of N-hydroxysuccinimide in 8 ml of DMF. After 24 hours of stirring at room temperature, the precipitated solid is filtered off and filtrate is cooled for 4 hours to 4C. The additional precipitated solid is again filtered off and the filtrate is liberated from the solvent in a vacuum. The residue is rewashed several times with ether and dried in a high vacuum.
Yield: 0.95 g (78~) H-NMR tCDCl3, ~, ppm):
6.80-6.20 (m, 6H, catechol); 4013 (t, 4H, OCH2CH2);
3.80 (s, 2H, OC(O)CH2N); 3.58 (t, 4H, NCHzCH2); 2.80 (s, 4H, C(O)CH2CH2C(0)); 2.07 (q, 4H, CH2CH2CH2) ., . ...~ . . , : - , ., ,. ~ , ,.,. . ~

` ~0~8~9 - 3~ -Exam~ o:
4-Nitrophe~yl-bi 9 ~ ( 3-(2,2-dimethyl-1,3-be~zodioxol-~-yloxy)propyl]amlne, 21 2.15 y (5 mmol) of 14 and 0.5 g (5 mmol) of triethylamine are introduced in 10 ml of ethanol and mixed with 1.41 g (10 mmol~ of 4-fluoronitrobenzene. The mixture is stirred for 3 days. Then, the solvent is removed and the crude product is purified chromatographically.
Yield: 1.50 g (55%) 1H-NMR (CDCl3, 5, ppm):
8.06 (d, 2H, nitrophenyl); 6.72 ~t, 2H, catechol); 6.71 (d, 2H, nitrophenyl); 6~47 (d, 2H, catechol); 6.46 (d, 2H, catechol); 4.13 (t, 4H, OCHzCH2); 3.67 (t, 4H, NCH2CH2); 2.11 (q, 4H, CH2CH2CH2); 1.~1 (s, 12H, C(CH3)2) 4-Ami~ophenyl-bist~3-~2,2-dimethyl-103-benzodioxol-4-yloxy)propyl]amine, 22 1.50 g (2.75 mmol) of 4-nitrophenyl-14 is dissolved in 30 ml of methanol and stirred with 150 mg of Pd/C (10~) under a hydrogen atmosphere for 4 hours at room temperature.
The catalyst is filtered off and the solvent is removed. A
greenish oil is obtained.
Yield: 1.27 g (89%) 4-Aminophenyl bis[3-~2,3-dihydroxyphe~oxy)propyl]aminohydrochlorid~, ~3 1.04 g (2 mmol) of 22, DIPACE is reacted under the conditions for the productiorl of 13 from 12. A white powder is obtained:
Yield: 0.965 g (94%) . .

i ~. .
.~

2 0 ~ 9 4-Isothioc~anato-bist3-(2,3-dihydroxyphe~o~y~propyl]aminohydroahloride, 24 0.51 g (1 mmol) of 23 of the preceding reaction ls reacted with 0.267 g (1.5 mmol) of N,N'-thiocarbonyldiimidazole. After completion of the reaction,imidazole is washed out with water. The product is obtained as yellow oil.
Yield~ 360 mg (70%) Example 11:
N,N-bist3-~2,2-Dimethyl-1,3-benzodioxol-4-yloxy)propyl~-N-~2,3-epoxypropyl)amine, 25 1.15 g (48 mmol) of sodium hydride is suspended in 40 ml of DMF at room temperature under nitrogen. A solution of 18.0 g of amine 14 (42 mmol) in 20 ml of DMF is slowly in-stilled in it and, after completion of the addition, it is stirred for 1 hour. Then, a solution of 5.48 g of epibromo-hydrin ~40 mm~l) in 20 ml of DMF is instilled and stirred for another`24 hours. The mixture is diluted with 70 ml of ice water, extracted several times with ethyl acetate and the combined organic extracts are dried on potassium carbo-nate. After removal of the solvent, a yellow oil remains.
Yield: 71%
H-NMR ~CDC13) 6.90-6.30 (m, 6H, catechol); 4.12 (t, 4H, OCH2CH2);
3.24 ~m, lH, epoxide); 2.80-2.60 (m, 8H, NC~z, epoxide); 1.93 (q, 4H, CH2CH2CH2); 1.69 (s, 12H, C(CH3) N,N-bis[3-~2,2-dimethyl-1,3-benzodio~ol-4-yloxy)propyl]~N
r2-hydroxy-3-(2-ni~roimidazolyl~propyl]amine, 26 A mixture of 2S0 mg of 2-nitroimidazole (2.2 mmol), 237 mg of 1,8-bis-(dimethylamino)naphthalene (1.1 mmol), 2.12 g of 25 (4.4 mmol~ and 5 ml of DMSO is heated with exclusion - . :., : . ~ . ,:
: , ..

2 Q ~

of moisture and stirring for 6 hours to 80C, the solvent is drawn off in a vacuum and the residue is chromatographed (silica gel, 230-400 mesh, 3 x 15 cm column, 20% CH3CN/CEICl3 to 8 0% CH3CH/CHCl3) . The individual fractions are examLned by thin-layer chromatography and thin~layer chromatography analogous fractions are combined. The solvent is drawn off and the residue is dried in a vacuum.
Yield: 27%
H--N~R (CDCl3) 8.40 (s, lH, imidazole); 8.34 (s, lH, imidazole); 6.90-6.30 (m, 6H, catechol); 4.20-4.00 (m, 5H, OCH2CHz, CHOH); 2. 90-2. 60 (m, 8H, NCH2CH2); 1. 93 (q, 4H, CH2CH2CHz); 1.69 (s, 12H, C(CH3)2) Example 12:
3-~N,N-biqt3-(2,2-Dimethyl-1,3-b~nzodio~ol-4-yloxy)propyl])ami~opropanoic acid ~thyl eYter, 27 0.898 g of potassium-tert-butylate (8.0 mmol) is dissolved in 150 ml of anhydrous tert-butanol and a solution of 27 . 24 g of 14 (llo mmol) in 30 ml of tert-butanol and 400 ml of ether are added. With stirring, 33.04 g of freshly distilled ethyl acrylate (330 mmol) is slowly distilled and the reaction mixture is left for 3 days at room temperature.
After removal of the solvent, the remaining oil is taken up in ether. The ether phase is washed neutral with water and dried on magnesium sulfate. After the concentration by evaporation, a pale yellow oil remains.
Yield: 48%
1H-NMF~ ( CDCl3 ) 6.90-6.30 (m, 6EI, catechol); 4.25-4.10 (m, 6H, OCH2CH2, CO2CH2CH3); 2.80-~. 50 (m, 8~, NCH2CH2, CH2N, COCH2); 1. 93 (q, 4H, CEl2CH2CHz); 1.69 (s, 12H, C(CH3)2); 1-25 (t, 3H, OCH2CH2 ) ~ . .. . . .
. . . .
:. .
:: :
.

- 41 - 2~8~

3-(N,N-biq[3-~2,2-Dimethyl-1,3-be~zodio~ol-4-yloxy}propyl])ami~opropanoic aci~ hydra~ide, 28 20 g of anhydrous hydrazine (624 mmol) is added to a solution of 10.0 g of ester 27 (18.9 mmol) in 200 ml of anhydrous pyridine and refluxed for 3 days. It is concentrated by evaporation to 5() ml and then mixed with 200 ml of water, extracted several times with ethyl acetate, the combined organic phases are washed with water~ dried on sodium sulfate and concentrated by evaporation. A white residue remains.
Yield: 69%
H-NMR (CDCl3) 6.90-6.30 (m, 6H, catechol); 4.08 (t, 4H, OCH2C~z);
2.80-2.50 (m, 8H, NCH2CH2, CH2N, COC~2~; 1.93 (q, 4H, CH2CH2CH2); 1.69 (s, 12H, C(CH3)z) Example 13:
3-~,N-hisr3-~2,2 Dimethyl-1,3-be~zodioxol ~-yloxy)propyl~aminopropanoic acid, 29 14.5 g of ester 27 (27.4 mmol) is refluxed in a solution of 5.00 g of potassium hydroxide (90.0 mmol) in 75 ml of 95% ethanol for 2 hours. The ethanol is drawn off in a vacuum and the remaining residue is taXen up in 100 ml of water. After shaking out with 50 ml of ether, the aqueous phase is carefully acidified with dilute hydrochloric acid.
The free acid is extracted by shaking out several tim~s with 50 ml of ether each. The combined ether phases are washed with saturated common salt solution and dried on magnesium sulfate. After removal of the solvent, a colorless oil remains.
Yield: 85 - : . '-.:, ' ,.' ~ . ' ' .' ;:,. .

~ ' :
, ,: .

2 0 ~

H-NMR (CDC13) 6.90-6.30 (m, 6H, catechol); 4.11 (t, 4H, OCH2CH2);
2.80-2.50 (m, 8H, NCH2CH2, CH2N, COC~2), 1.93 (q, 4H, CH2CHzCH2); 1.69 (s, 12H, C(CH3)2) 3-{N,N-bis[3-~2,2-Dimethyl-1,3-benzodio~ol-4-ylo~y)propyl]}aminosuccinimidopropionate, 30 The solution of 12.38 g of dicyclohexylcarbodiimide (60 mmol) in 50 ml of tetrahydrofuran is instilled in a solution, cooled to -5C, of 2S.1 g of carboxylie aeid 29 (50 mmol) and 5.75 g of N-hydroxysuccinimide (50 mmol) in 100 ml of anhydrous tetrahydrofuran within 20 minutes and is stirred ~or another 2 hours at this temperature and then for another 15 hours at room temperature. After adding 200 mieroliters of acetic aeid, it is stirred for another hour, then filtered and the residue is extracted twice with hot tetrahydrofuran. The combined filtrates are evaporated to dryness and the residue is recrystallized from ethyl aeetate.
Yield: 65~ -lH-NMR (CDCl3) 6.90-6.30 (m, 6H, catechol); 4.11 (t, 4H, OCHzCH2);
2.80-2.50 (m, 8H, NCH2CH2, CH2N, COCH2); 2.76 (s, 4H, COCH2CH2C0); 1.93 (q, 4H, CH2CH2CHz); 1-69 (s, 12H, C(CH3)2) E~am~le 14:
3-{NfN-bi~[3-~2,2-dimethyl-1,3-benzodioxol-4 yloxy)propyl]}aminosuccinimidopropionate, 30 The solution of 10.37 g of 1-(3-dimethylaminopropyl) 3-ethylearbodiimide (54 mmol) in 100 ml of acetonitrile is instilled in a solution, eooled to 0 C, of 25.1 g of earboxylie aeid 29 (50 mmol) and 8.30 g o.f 2,3,5r6-.

: ~4~9 tetrafluorophenol (50 mmol) within 5 minutes and heated for 2 hours to 75C. After adding 200 microliters of acetic acid, it is stirred for another hour, then filtered and the residue is extracted twice with hot acetonitrile. The combined filtrates are evaporated to dryness and the residue is recrystallized from ethyl acetate.
Yield: 65%
H-NMR (CDCl3) 6.90-5.30 (m, 7H, catechol, tetrafluorophenol); 4.11 (t, 4H, OCH2CHz); 2.80-2.50 (m, 8H, NCH2CH2, CH2N, COCH2); 1.93 (q, 4H, CH2CH2CH2); 1.69 (s, 12H, C(CH3)2 Exampl~ 15:
3-{~l,N-bi~3t3-(2,2-Dimethyl-1,3-}~enzodioxol-4-yloxy~propyl]}
aminopropa~ol, 32 15The solution of 20 g of ester 27 (38 mmol) in 50 ml of anhydrous ether is instilled in a suspension of 2.88 g of lithium aluminum hydrida (76 mmol) in 150 ml of anhydrous ether within one hour so that the solution boils moderately.
Then, it is refluxed for another 5 hours, cooled ta room temperature and excess hydride is carefully hydrolyzed with water. It is filtared off from precipitated hydroxide and the filtrate is washed several times with warm ether. After ~ -removal of the solvent in a vacuum, the residue is boiled up briefly in ethanol, filtered again and the solvent is drawn off. A highly viscous liquid remains.
Yield: 61%
1H~ ( CDC13 ) 6.gO-6.30 (m, 6H, catechol); 4.12 (t, 4H, OCH2CH2);
3.54 (t, 2H, CH20H); 2.80-2.60 (m, 6H, NCH2); 2.00-1.80 (m, 6H, C~2CH2CH2); 1-69 (s, 12H, C(CH3)2) ., :
:: . :. .
, . ~
' ' , , 2 ~ 9 Example ~6:
bis~3-~2,2-Dimethyl 1,3 benzodioxol-4-yloxy~propyl3-N-~3-chloropropyl)amine, 33 A solution of 10.1 g of alcohol 32 (20.7 mmol) in 50 ml of anhydrous carbon tetrachloride is mixed under a nitrogen atmosphere with 7.86 g of triphenylphosphine (30 mmol). It is refluxed for several hours. After cooling off, it is diluted with half the volume of petroleum ether and stored for some time at -20 C. The precipitate is suctioned off and washed with petroleum ether, after drying on sodium sulfate and removal of the solvent, a yellow oil remains.
Yield: 78%
H-NMR (CDCl3~
6.90-6.30 (m, 6H, catechol); 4.11 (t, 4H, OCH2CH2);
3.62 (t, 2H, CH~Cl); 2.80-2.60 (m, 6H, NCHz); 2.00-1.80 (m, 6H, CH2CH2CH2); 1.66 (s, 12H, C(CH3)2) Example 17:
3~N,N-bis r3- (2,2-Dimethyl-1,3-benzodioxol-4-yloxy1propyl]~aminopropanol, 34 4.88 g of alcohol 32 (10 ~mol), dissolved in 20 ml of dichloromethane, is added all at once to a well-stirred suspension of 3.23 g of PCC in 25 ml of anhydrous dichloromethane and the mixture is stirred for 90 minutes at room temperature. After adding 50 ml of anhydrous ether, it is decanted and the residue is washed three times with 20 ml each, the combined ether solutions are filtered on 20 g of silica gel. After removal of the solvent, a yellow oil remains.
Yield: ~0%

, . . .
' ,: . . " : :
- , . : . .::

2 0 ~
-- ~5 --1~--N2IR ( CDCl3 ) 9.74 (s, lH, aldehyde); 6.90-6.30 (m, 6H, catechol);
4 . 12 (t, 4H, OCH2CH2); ~ . 80-2 . 60 (m, 6H, NCH2); 2 . 42 (t, 2H, CH2CH0); 2.00-1.70 (m, 6H, CH2CH2CH2); 1.66 (s, 12H, C (CH3) 2 Example ~8:
3 r ~N,N-bis~3-~2-2-Dimethyl-1,3-be~zodio~ol-4-yloxy)propyl]~aminopropio~ic acid ~itrile, 35 0.90 g of potassium-tert~butylate (8.0 mmol) in 150 ml of anhydrous tert-butanol is dissolved and a solution of 27.2 g of 14 (110 mmol) in 30 ml of tert-butanol and 400 ml of ether is added. With stirring, 17.5 g of freshly distilled acrylonitrile (330 mmol) is slowly instilled and the reaction mixture is refluxed for 12 hours. After removal of the solvent, the remaining oil is taken up in ether. The ether phase is washed neutral with water and dried on magnesium sulfate. After the concentration by evaporation; a pale yellow oil remains. `~
Yield: 62%
H-NMR (CDC13) 6.90-6.30 (m, 6H, catechol); 4.11 (5, 4H, OCH2CH2);
2.81 (t, 4H, NCH2CH2); 2.50-2.30 (m, 4H, NCHzCH2CN);
1.97 (q, 4H, CH2CH2CH2); 1.69 (s, l~H, C(CH3)2) Example 19 N,N-bis~3-(2,2-Dimethyl-1,3-benzodio~ol-4-yloxy)propyl]propylenediaminff, 36 39 . 0 g of 100~ sulfuric acid (0.40 mol) is slowly instilled in a suspension of 30. 6 g of lithium aluminum hydride (0.81 mol) in 500 ml of anhydrous ether under ice cooling. Then, it is stirred ~or one hour at room 20l~8~9 temperature, then the solution of 12.55 g of nitri~e 35 (0.26 mmol) in 50 ml of anhydrouc; ether is instilled so that the solution boils moderately. Then, it is refluxed for another 8 hours, cooled to room temperature and excess hydride is carefully hydrolyzed with water. A solution of 40 g of NaOH in 360 ml of water is added, ~iltered off from the precipitated hydroxide and the filtrate is washed several times with warm ether. The combined ether extracts are dried on potassium carbonate and the solvent is drawn off. A yellow oil remains.
Yield: 47 H-NMR (CDCl3) 6~90-6.30 (m, 6H, catechol); 4.12 (t, 4H, OCH2CH2);
2.80-2.50 (m, 8H, NC~2); 2.00-1.80 (m, 6H, CH2CH2CH2);
151.69 (s, 12H, C(CH3)2) Example 20:
N~N-bisC3-~2,2 Dimethyl-1~3-benzodio~ol-4-yloxy)propyl]-N-t4-(nitro~enzyl)]amine~ 37 1.15 g (4~ mmol) of sodium hydride is suspended in 40 ml of DMF at room temperature under nitrogen. A solution of 18.0 g of amine 14 (42 mmol) in 20 ml of DMF is slowly instilled in it and after completion of the addition, it is stirred for another hour. Then, a solution of 8.64 g of 4-nitrobenzyl bromide (40 mmol) in 20 ml of DMF is instilled and it is stirred for another 24 hours. The mixture is diluted with 70 ml of ice water, extracted several times with ethyl acetate and the combined organic extracts are dried on potassium carbonate. After removal of the solvent, a yellow oil remains.
30Yield: 71%

, ~ - : , '.
.

2 ~ 9 - 4~ -H NMR ([D6]-acetone) 8.21 (d, 2H, nitroaryl); 7.53 (d, 2H, nitroaryl~; 6.60 6.40 (m, 6H, catechol); 4.10 (t, 4H, OC~2C~2); 3.63 (s, 2H, C6HsCH2N); 2.61 (t, 4H, NCH2CHz~; 1.91 (q, 4H, CH2CH2CHz); 1.60 (s, 12H, C(CH3)2 N,N-bis[3-~2,2-Dimethyl-103-benZodio~ol-4-yloxy)propyl]-N
(4-aminoben~yl)amine, 38 200 mg of 10% Pd/C in 250 ml of methanol is suspended in a 500 ml two-necked flask, cooled to -20C and saturated with water. Then, the solution of 5.0 g of 37 (8.8 mmol) in 50 ml of methanol is quickly instilled and stirred at -20C.
After completion of the absorption of hydrogen, it is separated from the catalyst and the solvent is drawn off in a vacuum. Pale yellow crystals remain.
Yield: 85%
H-N~R ( CD6]-acetone) 7.04 (d, 2H, aminoaryl); 6.60-6.40 (m, 8H, aminoaryl, catechol); 4.10 (t, 4H, OC~2CH2); 3.36 (s, 2H, C6H5CH2N);
2.63 (t, 4H, NCH2CH2); 1-89 ~q, 4H~ CH2CH2CH2); 1-59 (s, 12~, C(cH3)2 N~N-bis[3-~2~3-Dihydroxyphe~oxy)l?~
aminobenzyl)aminohydrochloride, 39 10.7 g (20 mmol) of 37 is dissolved in 80 ml of glacial acetic acid and mixed in boiling heat within two hours with 80 ml of an acid mixture (50% glacial acetic acid, 30%
water, 20% fuming hydrochloric acid). In doing so, solvent distills off with acetone that is being liberated as azeotrope. After completion of the addition, it is distilled for another 30 minutes. Then, residual solvent is drawn off on a rotary evaporator. The remaining residue is dried for 6 hours in a high vacuum.

, ,: : .: : : : ,,;.
, - 4~ -Yield: 79~
H-NMR ~[D5] pyridine) 8.93 (s, 6H, OH and NH2); 7.10-6.40 (m, 10H, aminoaryl, catechol); 3.92 (t, 4H, OCH2CH2); 3.47 (s, 2H, C6HsC~2N);
3.12 (t, 4H, NCH2CH2); 2.25 (q, 4H, CH2CH2CH2) N~N-bi:;r3~ 2-Dimethy~ 3-be~zodioxol-4-yloxy)pr t~-isothiocyanatobenzyl)ami~ohydrochloride, 40 1.15 g of thiocarbonyldichloride (10 mmol) is added to a solution of 1.10 g of aniline 39 (2.23 mmol) in 50 ml of lQ 3M hydrochloric a~-id and 50 ml of chloroform under a nitrogen atmosphere with a one-way spray and is intensively stirred for 6 hours at room temperature. Then, it is evaporated to dryness in a vacuum.
Yield: 78%
l_NMR ([D5]-pyridine) 8.96 (s, 4H, OH); 7.10-6.40 (m, 10H, aryl, catechol);
3.92 (t, 4H, OCH2CH2); 3.61 (s, 2H, C6HsCH2N); 3.21 (t, 4H~ NCH2CH2); 2 27 (q, 4H, CHzCH2CH2) Example 21:
N,N-bis[3-(2,2-Dimethyl-1,3-benzodioxol-4-yloxy)propyl]-N-2-propeD.yl ) amine, 41 1.20 g (50 mmol) of sodium hydride is suspended in 50 ml of DMF at room temperature under nitrogen. A solution of 20.6 g of amine 14 (48 mmol) in 40 ml of DMF is slowlv instilled in it and, after completion of the addition, it is stirred for another hour. Then, a solution of 6.04 g of allyl bromide ~50 mmol) in 25 ml of DMF is instilled and stirred for another 24 hours. The mixture is diluted with 70 ml of ice water, extracted several times with ethyl acetate and the combined organic extracts are dried on --`' 2~8g9 potassium carbonate. After removal of the solvent, a yellow oil remains.
Yield: 75%
H-NMR tCDCl3) 6.90-6.30 (m, 6H, catechol); 5.62 (m, lH, CH-CH2); 4.93 (m, 2H, CH=CH2); 4 . 09 (t, 4H, OCH2CH2); 2.80 2.60 (m, 6H, NCH2~; 2.32 (m, 2H, CH2CH=CH2); 2 . 00-1 . 80 (m, 6H, CEI2CX2CH2); 1.66 (s, 12H, C(CH3~2) Example 22:
bis[3-(2,2-Dimethyl-1,3-benzodio~ol-~-ylo~y~propylJ t2-propinyl)amine, 42 0.24 g (10 mmol) of sodium hydride i5 suspended in 50 ml of DMF at room temperature under nitrogen. A solution of 4.29 g of amine 14 (10 mmol) in 20 ml of D~F is slowly instilled in it and, after completion of the addition, it is stirred for another hour. Then, a solution of 1.43 g of propargyl bromide (12 mmol) in 10 ml of DMF is instilled and stirred for another 24 hours at 50C. After the cooling off, the mixture is diluted with 70 ml of ice water, extracted several times with ethyl acetate and the combined organic extracts are dried on potassium carbonate. After removal of the solvent, a yellow oil remains.
Yield: 66%
H--NMR ( CDCl3 ) :
6.90-6.30 (m, 6H, catechol), 4015 (t, 4H, OCH2CH2~;
2.80-2.60 tm, 8H, NCH2, CH2CC~); 2 . 00-1 . 80 (m, 7H, CH2CH2CH2, CH2CCH); 1. 65 ~s, 12H, C(CH3)2) 88~9 Example 23:
Coupling of a Tc-99m complex, aontaining isothiocyanate~, to prot~in~
The coupling of Tc-~9m complexes containing isothiocyanate (example 10) to proteins is to be described by the example of F(ab')z fragments of monoclonal antibody 17-lA. Instead of the antibody fragments, any other protein or a substance containing amino groups can be used.
Monoclonal antibody 17-lA is obtained corresponding to methods known in the literature after administration of 107 of the corresponding hybridoma cells in the abdominal cavity of a Balb/c-mouse and aspiration of the ascitic liquid after 7-10 days. The purification takes place according to methods also known in the literature by ammonium sulfate precipitation and affinity chromatography on protein A-sepharose. The purified antibody (10 mg/ml) is treated at pH 3.5 for 2 hours with 25 micrograms/ml of pepsin and the F(ab')2-fragments are then isolated by FPLC~ Before coupling with the chelating agent, the fragments are dialyzed at 4C for 12-24 hours from 0.1 M KH2P0~0.1 M
NaHC03, pH 8.5. The protein concentration is adjusted to 10 mg/ml. The complex containing NCS labeled analogously to example 6 is added in a molar ratio of 1:10 (complex:
protein) to the protein solution. For conjugate formation.
the mixture is incubated for 1 hour at 37C.

F.xample 24:
Biodistribution of a Tc-99m complex aoupled to fragme~ts of monoclo~al antibody 17-lA
The biodistribution of protein-bound Tc-99m complexes is to be described by the example of a conjugate with F(ab')2 fragments of monoclonal antibody 17-lA. The antibody, from which the fragments are obtained, recognizes `i ' ' ~ :: : ' : ' ' ' ' ~: , ~,: ' ~- , ,, : : . -' ~ . . , ,: ', .,:

an antigen, which is expressed by the human carcinomic cell line l'HT29". A control cell line, which also was obtained from a human carcinoma (MX-1), does not express this antigen. Isolated cells of both lines are administered subcutaneously to immunodeficient nude mice. A~ter the tumors have grown to a size of 300-800 mg, the mice are intravenously administered 20 micrograms of the complex, labeled with 200 microCi of Tc-99m, coupled to F(ab')2-fragments (example 11). The i~mune reactivity of the conjugates is determined in a parallel manner by the binding to an excess of intracellular antigen and is 75-80%. The biodistribution is determined 24 hours after administration of the conjugate by killing the animals, removing the organs and measuring the radioactivity in the organs. The following table represents the found amounts of radioactivity and shows a marked concentration of chelate in the antigen-positive tumor.

Or~an % of_the administered dose per gram_of tissue Spleen 0.4 Liver 1.1 Kidneys 2.8 Lung o.
~uscle 0.1 ;-Blood 0.6 MX-l 1.9 HT29 8.8 Example 25:
Biodistribution o~ a Tc-99m complex aontaining biotin 20 microliters of a commercially available streptavidin-coupled sepharose gel (corresponding to 20 - : . , .., .~ . . :.
:-. ., 2 ~ 9 micrograms of streptavidin) is administered to a 200 g rat in the muscle of the left hind leg. Then, about 30 minutes later, the intravenous administration of 5 micrograms of the complex containing biotin (example 8) labeled with 200 microCi of Tc-99m takes place according to example 6. The determination of radioactivity in the individual organs of the rat takes place after 4 hours. A 16-times higher radio-activity, which is found in the left hind leg muscle in com-parison with the right hind leg muscle, shows a clear spe-cific concentration of the Tc complex by binding to strept-avidin-sepharose. In all other organs, no activities over 1.4% of the administered dose per gram of tissue are de-tected after 4 hours. The highest concentration after the left hind leg muscle (1.4% of the administered dose per gram of tissue~ is found in the kidneys with 0.6% of the admini-stered dose per gram of tissue. About 89% of the admini-stered radioactivity is found in the urine after 4 hours.
The exampl~ shows that the complexes containing biotin in the organism can bind to streptavidin conjugates. In-stead of a streptavidin-sepharose conjugate, selective sub-stances, such as, e.g., monoclonal antibodies, enzymes or hormones, can be used, which -- coupled to streptavidin --can be detected after selective concentration in lesions or certain tissues by Tc-99m complexes containing biotin.
The preceding examples can be repeated with similar success by substituting the generically or specifically described reactants and/or operating conditions of this invention for those used in the preceding examples.
From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope t:hereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

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Claims (18)

1. A compound of formula I, (I) wherein X is -O-; -S-; -NR2-, wherein R2 is H; or a C1-6-alkylene radical, Y and Z are the same or different and are an -OH, -NHR3 or -SR3 radical, wherein R3 is H or a C1-6-alkyl radical, U is H or a branched or unbranched C1-6-alkyl, C1-6-alkoxy, hydroxyl or carboxyl radical, n is 2, 3, 4, 5 or 6, m is 2 or 3, and R1 is only present if m is 2, R1 is H, a benzyl radical or a branched or unbranched C8-6-alkyl radical, which alkyl radical is optionally substituted with 1, 2 or 3 hydroxyl, carboxyl or amino groups, and wherein said benzyl or alkyl radical optionally contains (i) a functional group -B, (ii) a compound -T which is capable of selectively concentrating in lesions or certain tissues, or (iii) a compound -T which is capable of selectively concentrating in lesions or certain tissues bound to the radical through a functional group -B-, wherein any optionally present functional group or a precursor thereof in R1 is optionally in protected form, when Y and/or Z are -NHR3, B is an amino, a hydrazino or hydrazide, a carboxyl, a C1-6-alkynyl or alkenyl, a hydroxyl, an amino-phenyl, an oxiranyl, a fluorinated phenoxycarbonyl or a biotin radical, or, when Y and Z are -OH or -SR3, B is as defined above, or a halogen, a formyl, a nitrile, a phenylisothiocyanato or a succinimid-oxycarbonyl radical optionally substituted with a sodium sulfate radical, and T is a monoclonal antibody or fragment thereof, a hor-mone, a growth factor, a ligand for a cell membrane receptor, a steroid, a neurotransmitter, a fatty aeid, a saccharide, an amino acid or oligopeptide, a biotin, or a radiosensitizer; and with the proviso that the compound is not N(CH2-CH2-CH2-O-C6H3-2,3-(OH)2)3;
or a technetium or rhenium complex thereof, or a salt of said compound or complex with an inorganic or organic acid.

2. A compound of claim 1, wherein U is H.

3. A compound of claim 1, wherein X is -O-.

4. A compound of claim 1, wherein n is 3.

5. A compound of claim 1, wherein Y and Z are the same and are -NH2 or -OH.

6. A compound of claim 1, wherein m is 2 and R is H, a benzyl radical, or an unbranched C0-3-alkyl radical which alkyl radical is optionally substituted with a hydroxyl or amino group, and wherein said benzyl or alkyl radical optionally contains a functional group -B, or a monoclonal antibody or fragment thereof, a steroid or misonidazole bound to the radical through a functional group -B-.

7. A compound of claim 1, wherein m and n are 3, X is -o-, U is H and Y and Z are the same and are NH2- or OH-.

8. A pharmaceutical preparation comprising an effec-tive amount of a technetium or rhenium complex of a compound of claim 1, and a pharmaceutically acceptable excipient.

9. A method of treating diseased tissue in a patient, comprising administering an effective amount of a technetium or rhenium complex of a compound of claim 1.

10. A method of claim 9, wherein the amount of the technetium or rhenium complex administered is 1 ? 10-5 to 5 - 104 nmol/kg of body weight.

11. A method of claim 9, wherein the amount of the technetium or rhenium complex administered is 5 to 500 mCi.

12. A method of claim 9, wherein the diseased tissue is a tumor.

13. A method of claim 9, further comprising first ad-ministering to the patient a diseased-tissue-specific agent coupled to streptavidin, and wherein the compound of claim 1 contains a compound -T which is biotin.

14. In a radiopharmaceutical diagnostic method, the improvement comprising administering to a patient an effec-tive amount of a technetium or rhenium complex of a compound of claim 1.

15. A method of claim 14, wherein the amount of the technetium or rhenium complex is 1 ? 10-5 to 5 ? 104 nmol/kg of body weight.

16. A method of claim 14, wherein the amount of the technetium or rhenium complex administered is about 0.05 to 50 mCi.

17. A method of claim 14, further comprising first ad-ministering to the patient a tissue-specific agent coupled to streptavidin, and wherein the compound of claim 1 con-tains a compound -T which is biotin.

18. A process for the production of a compound of formula I

(I) wherein X is -O-; -S-; -NR -, wherein R2 is H; or a C1-6-alkylene radical, Y and Z are the same or different and are an -OH, -NHR3 or -SR3 radical, wherein R3 is H or a C1-6-alkyl radical, U is H or a branched or unbranched C1-6-alkyl, C1-6-alkoxy, hydroxyl or carboxyl radical, n is 2, 3, 4, 5 or 6, m is 2 or 3, and R is only present if m is 2, R1 is H, a benzyl radical or a branched or unbranched C0-6 alkyl radical, which alkyl radical is optionally substituted with 1, 2 or 3 hydroxyl, carboxyl or amino groups, and wherein said benzyl or alkyl radical optionally contains (i) a functional group -B, (ii) a compound -T which is capable of selectively concentrating in lesions or certain tissues, or (iii) a compound -T which is capable of selectively concentrating in lesions or certain tissues bound to the radical through a functional group -B-, wherein any optionally present functional group or a precursor thereof in R1 is optionally in protected form, when Y and/or Z are -NHR3, B is an amino, a hydrazino or hydrazide, a carboxyl, a C1-6-alkynyl or alkenyl, a hydroxyl, an aminophenyl, an oxiranyl, a fluorinated phenoxycarbonyl or a biotin radical, or, when Y and Z are -OH or -SR , B is as defined above, or a halogen, a formyl, a nitrile, a phenylisothiocyanato or a succinimidoxycarbonyl radical optionally substituted with a sodium sulfate radical, and T is a monoclonal antibody or fragment thereofl a hormone, a growth factor, a ligand for a cell membrane receptor, a steroid, a neurotransmitter, a fatty acid, a saccharide, an amino acid or oligopeptide, a biotin, or a radiosensitizer; and with the proviso that the compound is not N{CH2-CH2-CH2-O-C6H3-2,3-(OH)2)3;
or a technetium or rhenium complex thereof, or a salt of said compound or complex with an inorganic or organic acid, comprising (a) reacting an amine of formula II, R1--N-{-(CH2)n--NU}m (II) wherein Nu is a nucleofuge and R1' is a substituent R1, wherein any optionally present functional group or a precursor thereof in R1 is present in protected form, and R1 contains no compound T, with an aromatic compound of formula III

(III) wherein U' is a substituent U
wherein, when U is a hydroxyl or carboxyl radical, said radical is present in protected form, and Y' and Z' are Y and Z, or a precursor thereof, or a protected form thereof, under base catalysis in polar solvents at temperatures of 50-200°C for about 2 hours to 6 days, (b) generating a functional group B optionally contained in R1 or a functional groups Y and Z, optionally, (c) coupling the thus obtained couplable or complexable compound with a compound T which is capable of selectively concentrating in lesions or certain tissues, and/or (d) complexing said compound with a technetium or rhenium isotope, or optionally, steps (c) and (d) can be performed in reverse order;
and (e) removing any remaining protective group or (f) convertiny any remaining precursor to the final product.