CATIONIC COMPLEXES OF TECHNETIUM-99M

Abstract: The invention concerns cationic complexes of Technetium-99m with bidentate ligands (L), including complexes having the formulae [Tc(NO) X L₂]⁺ and [Tc L3]⁺, which are of interest as heart imaging agents. The ligands are characterized by having the formula Y₂QZQY₂, where each Q is phosphorus or arsenic, Z is a 2 or 3 carbon atom linking group, and at least one of the groups Y and Z includes at least one -COC- ether group.

Description: This invention relates to cationic complexes of technetium-99 m (Tc-99 m) useful as body imaging agents. Radiopharmaceuticals may be used as diagnostic or therapeutic agents by virtue of the physical properties of their constituent radionuclides. Thus, their utility is not based on any pharmacologic action. Most clinically used drugs of this class are diagnostic agents incorporating a gamma-emitting nuclide which because of physical or metabolic properties of its coordinated ligands, localises in a specific organ after intravenous injection. The resultant images can reflect organ structure or function. These images are obtained by means of a gamma camera that detects the distribution of ionising radiation emitted by the radioactive molecules. The principal isotope currently used in clinical diagnostic nuclear medicine is metastable technetium-99 m (t½ 6 hours). It is well established that neutral bidentate ligands of the general type R₂Q(CH₂) n QR₂ (where Q may be phosphorus or arsenic, and n is 2) form stable well characterised cationic complexes with ⁹⁹Tc and 99 m Tc [1] . Several patents, including US 4481184, US 4387087, US 4489054, US 4374821, US 4451450 and US 4526776 describe various ligand species in which the co-ordinating atoms are phosphorus or arsenic, with predominantly alkyl and/or aryl substituents. The present invention concerns neutral bidentate donor ligands based on Phosphorus and which contain ether linkages; and cationic complexes of Tc-99 m comprising these ligands. It is found that such complexes show suprising properties which may make them superior body imaging agents, particularly heart imaging agents, to comparable complexes not containing ether linkages. In one aspect the invention is concerned with ligands having the formula: Y₂PZPY₂ where the groups Y may be the same or different and each is H or C1-C8 saturated hydrocarbon or saturated fluoro-hydrocarbon which may contain up to 3 ether oxygen atoms. Z is a _CC_ or _CCC_ or _COC_ chain which may be substituted by at least one C1-C8 saturated hydrocarbon or saturated fluorohydro-carbon group which may contain up to 3 ether oxygen atoms, provided that the ligand contains at least one _COC_ ether linkage. Preferably, the ligand contains 2 or 3 _COC_ ether linkages. Certain of these ligands are claimed as new compounds. Examples of C1-C8 saturated hydrocarbon groups optionally containing up to 3 ether oxygen atoms are Alkyl Cyclohexyl Alkoxy Alkoxyalkyl Alkoxyalkoxyalkyl Alkoxyalkoxyalkoxyalkyl _CH₂OC(CH₃)₂OCH₂_ _CH₂CH₂OCH₂CH₂_ 2-Tetrahydrofuryl Saturated fluorohydrocarbon groups may be, for example, any of the above in which one or more hydrogen atoms are replaced by fluorine atoms. The groups Y may be the same of different and each is preferably H or C1-C4 alkyl which may be substituted by C1-C4 alkoxy. Those groups Y which contain no ether oxygen atom are preferably H, methyl or ethyl. Z is preferably a _CC_ or _CCC_ or _COC_ chain which may be substituted by C1-C4 alkoxy or alkoxyalkyl or spirocyclic ether. More preferably Z is a _CC_ or _CCC_ chain, each Y is H or methyl, and either one or more methoxy substituents is attached to one or more groups Y, or one or more methoxymethyl or _COC_ spiro-cyclic ether groups is attached to a carbon atom of Z. In another aspect the invention provides a cationic complex of Tc-99 m with the ligand as defined. The cationic complex preferably has a formula selected from: [Tc(NO) n X m L₂]⁺A _ and [TcL₃]⁺A _ where X is a monodentate ligand for Tc A an is anion n is 1 or 2 and m is correspondingly 1 or 0, and L is the ligand. In the cationic complexes, X is a monodentate ligand for Tc, generally a halide such as F, Cl, Br or I, or a pseudohalide such as SCN, N₃, CN or RS. A is an anion whose nature is not critical, but which may conveniently be selected from the list given for X. The ligand is characterised by containing at least one _COC_ ether linkage. This may be provided by means of a _COC_ chain linking the two phosphorus atoms. More usually, the ether linkage is provided by means of an alkoxy or alkoxyalkyl substituent on one or more of the groups Y and Z. Preferably, Z is a _CC_ or _CCC_ chain, each Y is a H or methyl, and one or more methoxy or methoxymethyl substituents is attached to one or more groups Y or Z. Various substitution patterns are envisaged, for example: a) Backbone substitution which may be single or multiple for example: b) Phosphorus functionalisation which may be unsymmetrical or symmetrical for example: c) A mixture of functionalisation at the phosphorus atom and in the backbone. d) The spirocyclic type, for example: As indicated in the Examples below, these cationic complexes of Tc-99 m are of interest as heart imaging agents. The size of the groups Y and Z, and the number and size of the alkoxy, alkoxyalkyl or spirocyclic ether substituents, can be chosen to make the complex as lipophilic as may be desired for this purpose (or for any other purpose for which the complexes are to be used). But the ether linkages are not there merely to provide a desired hydrophilic/lipophilic balance. It has surprisingly been found that complexes formed from ligands with an ether linkage have significantly higher heart uptake than similarly hydrophobic complexes formed from corresponding ligands without an ether linkage. It appears that the ether linkages in the ligand modify the biologicial behaviour of the Tc-99 m complex, compared to the unsubstituted analogue, by providing increased blood and liver clearance which are essential to acheive good target organ to background ratios. Preferred cationic complexes technetium-99 m with bidentate ligands of this invention have formulae: a) [Tc(NO)XL₂]⁺A _ where X is a monodentate ligand for Tc, A is an anion, and L is a ligand having the formula b) [TcL₃]⁺A _ where L is a ligand having the formula Although this invention is concerned with results rather than with mechanisms, applicants offer the following as a possible explanation of mechanisms. Broadly, for compounds of similar structure, there is a relationship between lipophilicity and protein binding. Compounds of high lipophilicity are more strongly bound to proteins than compounds of low lipophilicity. For 99 m Tc cations, the effect of high protein binding is that they remain a long time in circulation, so that the image of the heart muscle at convenient imaging times post-injection is obscured by the blood pool activity. A further generally observed tendency for the more highly lipophilic cations is that they possess slow clearance through the hepatobiliary system, so that heart imaging can be impaired by liver activity. Substantially increasing the hydrophilicity of a 99 m Tc complex has the desired effect reducing protein binding but also reduces heart uptake. It appears that there is, however, a region of intermediate lipophilicity where the heart uptake is retained and there is also absent, or sufficiently weak, protein binding to permit rapid clearance from blood. It appears that there may be a ranking in contribution to polarity of TcO greater than MeO greater than EtO. By means of this background understanding, it may be possible to achieve the required lipophilic/hydrophilic balance through the additive hydrophilic effects of the oxygen substituents balancing out the lipophilic effects of the hydrocarbon moietes in the molecule. These phosphine ligands are not easy to make. They are toxic, the compounds or their precursors often spontaneously flammable in air, and their preparation frequently entails hazards of explosion or difficult to control reactions. Conditions may need to be carefully chosen to avoid the risk of side-reactions. The following reaction schemes are available: This route is available for other symmetrical ligands which are mono-substituted at each P atom. This route is available for other unsymmetrical ligands which are mono-substituted at one P atom. This route is available for other ligands where the backbone is substitued by alkoxy or alkoxyalkyl or spirocyclic ether. These and other routes are described in more detail in Examples 1, 2 and 8 to 12 below. The cationic complexes of Tc-99 m may be prepared by methods well known in the art. For example: Complexes of the type [TcX₂L₂]⁺A _ may be prepared by methods as described in US Patents 4387087 and 4489054. Tc(VII)-pertechnetate is reduced to a lower valency state in the presence of the ligand. If the ligand or the final Tc-99 m complex is not water soluble, it may be necessary to carry out the complexation reaction in a biphasic system, in which the technetium starts in the aqueous phase and is extracted into a non-aqueous phase during the preparation. The complexation reaction either in monophasic or biphasic systems, is carried out so that a large stoichiometric excess of lipophilic ligand is utilised over the stoichiometric amuont of Tc-99 m plus Tc99, present in the pertechnetate state or reduced state. Preferably, more than a tenfold excess of lipophilic ligand over 99 m-Tc + 99Tc is used. The reaction is carried out over a wide range of temperatures, with a proviso that the aqueous phase (and non-aqueous phase, if used) should remain liquid throughout the preparation. Thus, temperatures in the range of 0-80°C are normally used, most preferably 10-50°C. The reaction times range from a few minutes to 1-2 hours. It should be recalled that the half-life of Tc-99 m is only 6 hours and the reaction times have to be adjusted accordingly. Complexes of the type [Tc(NO) n X m L₂]⁺A _ may be prepared by methods as described in EP-A-0291281 which falls under Article 54(3) EPC. a) Reaction of the 99 m Tc-NO compounds described with a suitable ligand. Reaction conditions depend on the nature of the ligand, but typically involve holding the reactants at ambient or elevated temperature in an aqueous medium. b) Reaction of suitable preformed 99 m Tc radio-pharmaceutical complexes with hydroxylamine salts. This can result in exchange of e.g. halogen, oxo (mono or bis oxo complexes), phosphine or isonitrile ligands for nitrosyl groups. Again, reaction conditions depend on the particular ligand involved, but generally involve holding the reactants at ambient or elevated temperatures in an aqueous medium. c) A one-step preparation, in which pertechnetate, e.g. from a technetium generator, is added to a mixture of a hydroxylamine salt and the chosen ligand. In this reaction, the hydroxylamine salt is acting as a reducing agent. As compared with stannous ion and other reducing agents, hydroxylamine salts have the advantage, as previously noted, of being able to reduce technetium to a low oxidation state provided that the ligand is capable of stablising technetium in that state. As before, the reaction conditions depend on the nature of the ligand, but typically they involve holding the reactants at ambient or elevated temperature in an aqueous medium. Complexes of the type [TcL₃]⁺A _ may be prepared by methods as described in US Patent 4481184. Pertechnetate ion (in the form of a salt) is reduced by heating in the presence of a liquid (L). The reduction can be carried out in the presence of a chemical reducing agent, e.g., sodium borohydride, stannous chloride, or sodium hyposulphite. The solvent system used for the reaction is not critical; methanol, ethanol or a combination of either with water is preferred. If a chemical reducing agent (other than the ligand) is not present, the reaction will preferably be carried out in a pressure vessel at a temperature greater than about 100°C. If an added chemical reducing agent is used, the reaction will preferably run under reflux conditions. In addition to pertechnetate, ligand, chemical reducing agent (optional) and solvent, the reaction mixture will also contain the anion "A" in the form of a salt. In carrying out the above described reduction reaction the molar ratio of ligand (L) to pertechnetate ion should be not less than about 6: 1. The larger the excess of ligand used, the more carrier-free the product. The concentration of anion (A) is not critical. However, the anion will preferably be chlorine, present as sodium chloride in an amount sufficient to provide an isotonic concentration of 0.15 molar after the pertechnetate ion has been added. The following Examples illustrate the invention Synthesis of Methylether Substituted Diphosphines All reactions and manipulations were performed under vacuo or oxygen free nitrogen atmospheres. Solvents were dried and degassed by nitrogen purge or freeze/thaw cycles prior to use. The reagents BrCH₂OMe and ClCH₂OMe were purchased or obtained from Amersham (BrCH₂OMe and ClCH₂OMe were distilled prior to use) and were deoxygenated by freeze thaw cycles. MeLi was prepared from MeCl in diethylether and was estimated and used in diethylether solution. Ammonia was dried by distillation from sodium. The phosphines MePH-(CH₂)₂-HPMe [Ref. 1], and Me₂P-PMe₂ [Ref. 2] were prepared according to established procedures. Me₂P(CH₂)₂P(H)Me [Ref. 3] was prepared from Me₂P(CH₂)₂PH₂ [Ref. 4]. Abreviations used are: THF = tetrahydrofuran; R. T. = ambient temperature; ether = diethylether = Et₂O; Me = methyl. Example 1 Procedure: To a 250 ml 3 neck round bottom flask equipped with a condenser, a pressure equalising dropping funnel and a magnetic stirrer, was transferred the phosphine MePH-(CH₂)₂-PHMe, ((6.76 g; 5.53 × 10 _2 moles) in 35 ml THF. MeLi (11.67 × 10 _2 moles in ether solution) was taken into the dropping funnel and added dropwise to the phosphine with stirring at room temperature, which changed colour to light yellow. The dropping funnel was rinsed with 10 cm³ THF and charged with BrCH₂OMe (13.83 g, 11.07 × 10 _2 moles) in 20 ml of THF. The reaction flask was cooled to _78°C and the solution of BrCH₂OMe was added dropwise with stirring, stirred at this temperature for 1 h, and allowed to warm to room temperature. The resulting suspension was hydrolysed, the organic layer separated and dried over magnesium sulphate overnight. The dried organic layer was distilled. As the temperature was raised gradually, ether together with the THF distilled at 38-70°C. Finally, the product distilled at 74-82°C under dynamic vacuum (about 0.1 mmHg) as a colourless liquid. Yield = 2.96 g (25.5%). Example 2 Procedure 1: In a 250 ml 3 neck round bottom flask, equipped with a dry ice condenser (with a T-adapter at the top as the N₂ inlet-outlet system), a pressure equalising dropping funnel and a magnetic stirrer, was condensed about 150 ml anhydrous liquid ammonia. The liquid ammonia was kept at _78°C and the phosphine Me₂P-(CH₂)₂-PHMe (2.06 g; 1.51 × 10 _2 moles) was transferred into the reaction flask. Required amount of MeLi (1.51 × 10 _2 moles) was added dropwise to the phosphine from the dropping funnel, which produced a deep orange colour. The dropping funnel was rinsed with 10 ml ether and charged with a solution of ClCH₂OMe (1.22 g, 1.51 × 10 _2 moles) in 20 ml ether. This solution was added dropwise to the reaction mixture until the deep orange colour disappeared. The reaction mixture was allowed to warm to room temperature and after the ammonia had evaporated, about 50 ml ether was added and the resulting slurry hydrolysed. The ether layer was removed, dried over magnesium sulphate overnight. The dried ether layer was distilled and after removing ether, the product distills at approx. 80°C under dynamic vacuum (0.1 mmHg) as a colourless liquid. Yield = 0.61 g (22.5%) Procedure 2 Note: As the starting phosphine Me₂P-(CH₂)₂-PHMe was contaminated with a small amount of Me₂P-(CH₂)₂-PMe₂, it was purified by precipitation of the salt Me₂P-(CH₂)₂-PMeLi by reacting with MeLi in ether, which would keep Me₂P-(CH₂)₂-PMe₂ in solution and enable easy removal. In a 500 ml 3 neck flask equipped with dry ice condenser (with a T adapter at the top as N₂ inlet-outlet system), a pressure equalising dropping funnel and a magnetic stirrer, was transferred the phosphine Me₂P-(CH₂)₂-PHMe (4.9 g; 3.6 × 10 _2 mole) in 30 ml ether. MeLi (3.6 × 10 _2 moles in ether) was added dropwise from the dropping funnel to the phosphine solution. As there was no immediate precipitation, most of the ether was evaporated when precipitation occured. The remaining solution was filtered off, and the precipitate was washed twice with 10 ml portions of ether. The reaction flask was then cooled to _78°C and about 200 ml anhydrous liquid ammonia was condensed to the reaction flask. The precipitated colourless solid forms an orange slurry in contact with liquid ammonia. The dropping funnel was rinsed with 10 ml portions of ether and charged with a solution of ClCH₂OMe (2.9 g; 3.60 × 10 _2 moles) in 20 ml ether. This solution was added dropwise to the stirred orange slurry until the colour just disappears. The reaction mixture was allowed warm to room temperature and after all the ammonia has evaporated, about 50 ml ether was added and the resulting slurry was hydrolysed. The ether layer was separated, dried over magnesium sulphate overnight. The dried ether layer was distilled and after removing ether, the product distills at about 80°C/dynamic vacuum (0.1 mmHg) as a colourless liquid. As the product contained traces of water, it was dried over KOH for 4 h and redistilled. Amount = 1.7 g (26%) References 1. M. Baake, O. Stelzer and V. Wray, Chem. Ber., 113 , 1356 (1980); or by addition of MeI (2 mole equivalents) to H₂P(CH₂)₂PH₂ in methanol or ethanol followed by isolation of the disphosphonium salt [Me₂P(H₂)-(CH₂)₂-P(H₂)Me]²⁺.2I _ and liberation of the free disecondary phosphine by neutralisation. 2. D.W. Meek et al., Inorg. Synth., 14 , 15 (1973). 3. Prepared by deprotonation of Me₂P(CH₂)₂PH₂ with either MeLi or Bun n Li in ether or hydrocarbon solvents followed by formation of the required tertiarty-secondary phosphine by addition of MeI. 4. R.B. King and J.C. Cloyd, J. Amer. Chem. Soc., 97 , 46 (1975). R.C. Taylor and D.B. Walters, Inorg. Synth., 1973, 14 , 10. ¹H NMR data (360 MHz) ¹H NMR spectra were recorded on a Bruker WH360 operating at 360 MHz and were referenced on the protio impurity of the C₆D₆ used as solvent (7.27 ppm). B. Synthesis and Properties of Cationic Technetium-99 m Complexes of Ligands of Examples 1 and 2 Example 3 [TcX₂L₂]⁺ Technetium III disphosphine dihalide complex. Materials 8 mg EGTA 120 mg NaCl 3 mg FeCl₃/6µl L/1 ml EtOH 1.1 ml 99 m -TcO₄ _ Na generator eluate (at 1.49 GBq/ml) These constituents were placed in a sealed glass vial under N₂ and heated at 120°C for 70 minutes, to which 3 ml of 50% EtOH = saline, and 0.15 ml 1 M KHCO₃ were added, final pH = 6. The resulting solution was submitted to various analytical techniques, summarised below. Chromatography Data The resulting solution (above) contains no free 99 m TcO₄ _ and no colloid, and indicates that the technetium complex is present in solution in approximately 95% yield. Saline rf = 50.88 Methylethylketone rf = + 0.76 Acetonitrile/water 50: 50 rf = 50.80 (broad) Gel Electrophoresis Data The complex moved as a single band towards the cathode rf = _ 0.53 (_ indicating movement towards the cathode). HPLC Data The compound elutes as a single band with a retention time of approximately 7.0 minutes. EXAMPLE 4 Materials NH₂OH · HCl 2.5 mg SnF₂ 0.8 ml at 6.6 × 10 _5 M solution (aq.) L 5 µl 99 m TcO₄Na generator eluate at 316 mCi/ml Saline 0.4 ml Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 1 hour. The crude preparation was then subjected to HPLC purification, and the major component (retention time 6.3 minutes) collected, any remaining THF was removed and the resulting solution analysed in the usual way. A sample of this material was submitted for animal biodistribution studies, final pH = 7.4. Chromatography The resulting preparation contains no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 85% yield. Saline rf = 0.02 Methylethylketone rf = 0.69 Acetonitrile: Water, 50: 50 rf = 1.0 Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.75 (_ indicates movement towards cathode). HPLC Data The complex elutes as a single band with a retention time of approximately 6.3 minutes. Animal Biodistribution Data See Table I. EXAMPLE 5 Materials Ethanol 2 ml NaOH 0.05 ml, 10 M aq. solution Saline 0.6 ml L 20 µl 99 m TcO₄Na Generator Eluate 0.4 ml at 3.73 GBq/ml Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 30 minutes. This solution was then diluted with 3 ml 66% saline/ethanol, and the pH adjusted to 6 with 0.1 M HCl. The resulting solution was then submitted to chromatographic analysis and animal biodistribution study. Chromatographic Data The resulting preparation contains no free TcO₄ _ or reduced technetium colloid, and indicates that the desired species is present in solution approximately 90% yield. Saline rf = 0.00 Methylethylketone rf = 0.75 (broad) Acetonitrile: Water 50: 50 rf = 0.98 HPLC Data The complex elutes as a sharp band at approximately 8.1 minutes (∼ 10% minor component at ∼ 6 minutes = 10%). Animal Biodistribution Data See Tables II and III Example 6 Materials NH₂OH · HCl 25 mg SnF₂ 1.0 ml. 8 µg/ml L 10 µl 99 m TcO₄Na 1.2 ml at 243 mCi/ml (generator eluate) Method The components were mixed in a sealed N₂ purged vial and heated at 120°C for 1 hour. The crude preparation was then subjected to HPLC purification and the major component (retention time 6.5 minutes) collected, any remaining THF was removed and the resulting solution analysed in the usual way. A sample of this material was submitted for animal biodistribution studies, final pH = 7.4. Chromatography The resulting preparation contains no reduced technetium colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 95% yield. Saline rf = 0.04 Methylethylketone rf = 0.86 Acetonitrile: water 50: 50 rf = 0.90 Gel Electrophoresis Data The complex moves as a single band towards the cathode rf = _ 0.78 (_ indicates movement towards the cathode). HPLC Data This complex elutes as a single band, with a retention time of approximately 6.5 minutes. Animal Biodistribution Data See TabIe IV Example 7 Materials Ethanol 1 ml NaOH 0.05 ml, 10 M aq. solution Saline 2 ml L 10 µl 99 m TcO₄Na Generator Eluate 0.5 ml at 2.66 GBq/ml Method The components were mixed in a sealed N₂ purged vial and heated at 120°C for 30 minutes; pH of the solution was adjusted to 7-8 with 0.1 M HCl. The resulting solution was them submitted to chromatographic analysis and animal biodistribution study. Chromatography The resulting preparation contains no free TcO₄ _ or reduced technetium colloid, and indicates that the desired species is present in approximately 95% yield. Saline rf = 0.00 Methylethylketone rf = 0.67 Acetonitrile: water 50: 50 rf = 0.75 HPLC Data This complex elutes as a sharp peak at approximately 8.4 minutes (5% minor components between 4.5 to 6 minutes). Animal Biodistribution Data See Tables V and VI Comparative Data The bidentate phosphine ligand di-(dimethylphosphino) ethane (dmpe) is well known, and various Tc-99 m complexes have been proposed for use as heart imaging agents. The following Tables VII, and VIII compare the biodistribution data of two such prior art complexes with the data given above in respect of the complexes of Example 4, 5, 6 and 7 (derived from the ligands of Example 1 and 2). The Tables permit the following conclusion. From Table VII, the complex formed from the ligand of Example 1 shows: i) significantly higher heart uptake and retention at 2′ and 60′ ii) reduced liver retention at 60′ iii) significantly increased heart/liver ratio at 60′. From Table VIII, the complex formed from the ligand of Example 1 shows: i) increased heart uptake and retention at 2′ and 60′ ii) much lower liver retention at 60′ iii) better heart/blood, heart/muscle and heart/liver ratios at 60′. Complexes formed from the ligand of Example 2 show the same general trend. C. Synthesis of Backbone-Substituted Diphosphine Ligands EXAMPLE 8 Preparation of Bis(dimethylphosphinomethyl)ether. i. Bis(bromomethyl) ether This was prepared by the method of Stephen et al, J. Chem.Soc. , 1920, 117 , 515. ii. Bis(dimethylphosphinomethyl) ether To a solution of dimethylphosphine (5g) in diethyl ether (110 cm³) at _70°C was added portions of a solution on n-butyl lithium (2.5 M in hexanes) until formation of the phosphide anion was complete (as monitored by ³¹P nmr spectroscopy). The mixture was stirred for 10 minutes at _70°C and bis(bromomethyl) ether (5 g) then added. The mixture was allowed to warm to room temperature and then stirred at this temperature for several hours. The resulting mixture was extracted with portions of dilute hydrochloric acid (2 M) and the combined aqueous extracts washed with diethyl ether. The aqueous layer was slightly basified with aqueous sodium hydroxide (30%) and then extracted with chloroform. Removal of the solvent from the chloroform layer under reduced pressure (40°C at 150 mmHg) gave the diphosphine (2.8 g) in a substantially pure state. δ³¹P (CDCl₃) _ 51.1 δ¹³C (CDCl₃) 10.0(d, J = 12), 74.2(dd, J = 11, 5) EXAMPLE 9 Preparation of 1,3-Bis(dimethylphosphino)-2,2-bis(methoxymethyl)propane i. 1,3-Dibromo-2,2-bis(methoxymethyl)propane To 2,2-bis(bromomethyl)propane-1,3-diol (100 g) was added potassium bicarbonate (80 g) and dimethyl sulphate (170 g). The mixture was stirred and heated to 100°C. After about 30 minutes at this temperature the reaction became very vigorous (Care!). After a further 2 hours heating and stirring the mixture had become very viscous. The mixture was cooled, made slightly basic by the addition of sodium bicarbonate, and then extracted with chloroform (4 × 200 cm³). The chloroform extracts were combined and the solvent removed under reduced pressure to give the crude product. Repeated distillation under reduced pressure gave the pure product (25 g) as a colourless liquid (b.p. 65-70°C at 0.1 mmHg). δ¹³C (CDCl₃) 34.9(×2), 43.7, 58.9(×2), 71.0(×2) ii. 1,3-Bis(dimethylphosphinyl)-2,2-bis(methoxymethyl) propane To a stirred solution of dimethylphosphine (24 cm³) in dry diethyl ether (150 cm³) at _40°C was added a solution of n-butyl lithium (44 cm³, 2.5 M in hexanes). After about 30 minutes the temperature had risen to _20°C and 1,3-dibromo-2,2-bis(methoxymethyl)propane (15 g) was added, causing an exothermic reaction. The reaction mixture was allowed to warm to room temperature and then left stirring overnight. The ether solution was extracted withseveral portions of dilute hydrochloric acid (2 M), the combined aqueous extracts basified with aqueous sodium hydroxide, and the liberated phosphines extracted with chloroform. Nmr spectroscopy showed the formation of both the required diphosphine and a substantial quantity of 2,2-bis(methoxymethyl)propyldimethylphosphine. Addition of excess aqueous hydrogen peroxide (6%) converted both phosphines to their corresponding oxides. The aqueous layer was then separated and evaporated under reduced pressure. The resulting viscous oil was purified by chromatography on Florisil using methanol/ethyl acetate mixtures as eluants. The pure 1,3-bis(dimethylphosphinyl)-2,2-bis(methoxymethyl)propane (2.8 g) was isolated as a semicrystalline material. δ³¹P (CDCl₃) 41.8 δ¹³C (CDCl₃) 19.4(d, J = 69), 19.45(d, J = 69), 33.9(dd, J = 68, 5), 43.1(t, J = 5) 58.7(s), 58.7(s), 75.5(t, J = 8) iii. 1,3-Bis(dimethylphosphino)-2,2-bis(methoxymethyl) propane The previously prepared bis(phosphine oxide) (2.7 g) was added to lithium aluminium hydride (3.0 g) in dioxan (200 cm³) and the mixture heated under reflux for 2.5 hours. After cooling, the mixture was hydrolysed by the careful addition of aqueous dioxan (25 cm³, 1: 1), and then aqueous sodium hydroxide (4 cm³, 50%) was added. The resulting mixture was filtered and the bulk of the solvent then removed under reduced pressure (60°C at 100 mmHg) to give the diphosphine. ³¹P nmr showed no presence of other phosphorus-containg components. δ³¹P (CDCl₃/dioxane) _ 63 δ¹³C (CDCl₃) 15.7(m), 38.3(dd, J = 16, 10), 42.2(t, J = 10), 58.7(s), 76.7(t, J = 9) The diphosphine was converted to its crystalline disulphide derivative for analysis. Found: C, 41.9; H, 8.3. C₁₁H₂₆O₂P₂S₂ requires C, 41.75; H, 8.3%. EXAMPLE 10 Preparation of 1,3-Dimethoxy-1,3-bis(dimethylphosphino) propane i. 1,3-Dichloro-1,3-dimethoxypropane To a stirred quantity of 1,1,3,3-tetramethoxypropane (10 g) was cautiously added thionyl chloride (10 g) over a period of 10 minutes. The reaction was then stirred at room temperature until the reaction was complete (as indicated by nmr specttroscopy). The excess thionyl chloride was removed under reduced pressure (20 mmHg) and the residue distilled to give the dichloride (7.7 g) (b.p. 40°C at 5 mmHg). Nmr spectroscopy showed the presence of approximately equal quantities of two diastereoisomers. δ¹³C (CDCl₃) 48.3(×2), 48.7(×2), 57.5(×4), 96.2(×2), 96.4(×2) This material was found to deteriorate on standing and it was therefore prepared immediately prior to use. ii. 1,3-Dimethoxy-1,3-bis(dimethylphosphino)propane To a stirred solution of dimethylphosphine (18 cm³) in hexane (150 cm³) at _50°C was added a solution of n-butyl lithium (37 cm³, 2.5 M in hexanes). As the phosphide anion precipitated the mixture became very viscous and difficult to stir. After about 15 minutes 1,3-dichloro-1,3-dimethoxypropane (7.7 g) was added and the mixture allowed to warm to room temperature. The viscous mixture was agitated by bubbling dry nitrogen and then allowed to stand overnight. The resulting mixture was extracted with portions of dilute hydrochloric acid (2 M), the combined aqueous extracts basified with aqueous sodium hydroxide solution, and the liberated phosphines extracted into chloroform. Removal of the chloroform under reduced pressure gave a mixture of the two diastercoisomers of 1,3-dimethoxy-1,3-bis(dimethylphosphino)propane in a satisfactory yield. δ³¹P (CDCl₃) _ 43.6 and _ 44.6 δ¹³C (CDCl₃) 8.2(d, J = 14), 9.2(d, J = 14), 9.4(×2)(d, J = 14), 32.6(t, J = 13), 32.8(t, J = 13), 58.7-59.0(m), 79.0(dd, J = 10, 8), 80.0(cc, J = 12, 9) EXAMPLE 11 Preparation of 1,3-bis(dimethylphosphino)-2-(2′-methoxyethoxymethyl)-2-(methoxymethyl)propane i. 5,5-Bis(bromomethyl)-2-ethanol-1,3-dioxane 2,2-Bis(bromomethyl)propane-1,3-diol (250 g) and benzaldehyde (110 g) were added to benzene (500 cm³) containing concentrated sulphuric acid (2 cm³) and the mixture boiled. Water was removed using a Dean Stark apparatus. After about 6 hours the reaction was complete. The mixture was cooled and the sulphuric acid neutralised by the addition of excess sodium bicarbonate. The resulting solution was filtered and the benzene removed under reduced pressure to give an oil which solidified on standing. Recrystallisation from petroleum ether gave the 1,3-dioxane (296 g) as a white solid (m.p. 66°C). δ¹³C (CDCl₃) 34.4, 36.0, 37.2, 71.7(×2), 102.1, 125.9(×2), 128.2(×2), 129.2, 137.2. ii. 5-Bromo-5-(2′-methoxyethoxymethyl)-2-phenyl-1,3-dioxane Sodium (3.3 g) was added to 2-methoxypropanol (50 cm³) and the mixture was then stirred and warmed until the metal had dissolved. 5,5-Bis(bromomethyl)-2-phenyl-1,3-dioxane (50 g) in 2-methoxypropanol (150 cm³) was then added. The mixture was then boiled under reflux for 24 hours in an atmosphere of dry nitrogen. The excess 2-methoxypropanol was removed under reduced pressure and diethyl ether (550 cm³) added. The sodium bromide was filtered off and the ether removed under reduced pressure to give the product as an oil (49.3 g, 95%). This material was sufficiently pure to be used in the subsequent reaction without further purification. δ¹³C (CDCl₃) 36.1, 38.3, 58.8, 70.7(×2), 70.9, 71.1, 71.6, 101.8, 125.8(×2), 128.0(×2), 128.8, 137.7. iii. 5-(2′-Methoxyethoxymethyl)-5-(methoxymethyl)-2-phenyl-1,3-dioxan Sodium (1.4 g) was dissolved in dry methanol (20 cm³) and placed in a Teflon-lined autoclave (Berghof, 150 ml) together with 5-bromo-5-(2′-methoxyethoxymethyl)-2-phenyl-1,3-dioxane (20 g) in dry methanol (30 cm³). The reaction mixture was then heated at 150°C for 6 days. The solvent was removed from the reaction mixture under reduced pressure and the organic residue taken up into ether. Sodium bromide was filtered off and the ether removed under reduced pressure to give the crude product (17 g). This material was sufficiently pure to be used without purification. δ¹³C (CDCl₃) 38.7, 58.8, 59.1, 69.8(×2), 70.9, 71.0, 71.2, 71.6, 101.5, 125.9(×2), 128.1(×2), 128.7, 138.2. iv. 2-(2′-Methoxyethoxymethyl)-2-(methoxymethyl)propane-1,3-diol 5-(2′-Methoxyethoxymethyl)-5-(methoxymethyl)-2-phenyl-1,3-dioxan (17 g) was heated under reflux for 4 hours with a mixture of ethanol (130 cm³), water (50 cm³) and concentrated sulphuric acid (1.5 cm³). The mixture was then cooled, neutralised by the addition of sodium bicarbonate, filtered and the volume reduced to about 50 cm³ under reduced pressure. The aqueous residue was extracted with methylene chloride and the organic layer then dried over anhydrous sodium sulphate. Removal of the volatile components under reduced pressure gave an oil (9.5 g) which was shown to be largely the required diol. This was used without further purification. δ¹³C (CDCl₃) 44.9, 58.8, 59.4, 64.4(×2), 70.6, 71.5, 72.1, 74.3. v. 1,3-Dichloro-2-(2′-methoxyethoxymethyl)-2-(methoxymethyl)propane 2-(2′-Methoxyethoxymethyl)-2-(methoxymethyl)propane-1,3-diol (5.5 g), triphenylphosphine (24 g) and dry carbon tetrachloride were heated under reflux in an atmosphere of dry nitrogen until the reaction was complete (about 2.5 hours). The mixture was cooled, filtered and the solid residue washed with carbon tetrachloride. The carbon tetrachloride solutions were combined and the solvent removed under reduced pressure. Petroleum ether (250 cm³, b.p. 40-60°C) was added to the residue and the resulting suspension filtered. Removal of the petroleum ether under reduced pressure gave the crude product (6 g) as an oil. The dichloride was isolated by chromatography on Kieselgel 60 (Merck) using ethyl acetate/petroleum ether mixtures as eluants. Final purification was achieved by distillation under reduced pressure. The pure dichloride (1.4 g) was isolated as an oil (b.p. 97°C at 0.06 mmHg). δ¹³C (CDCl₃) 44.7(×2), 45.7, 58.8, 59.2, 68.9, 70.4, 70.8, 71.6. vi. 1,3-Bis(dimethylphosphino)-2-(2′-methoxyethoxymethyl)-2-(methoxymethyl)propane To a stirred solution of dimethylphosphine (5 cm³) in dry liquid ammonia (80 cm³) was added a solution of n-butyl lithium (10 cm³, 2.5 M in hexanes). This mixture was stirred at _60°C for 10 minutes and 1,3-dichloro-2-(2′-methoxyethoxymethyl)-2-(methoxymethyl)propane (1.4 g) in a little dry diethyl ether was then added. The resulting mixture was stirred for 15 minutes at _60°C and then allowed to warm up slowly to room temperature. When the ammonia had evaporated diethyl ether (150 cm³) was added and the resulting mixture extracted with portions of dilute hydrochloric acid (2 M). The combined aqueous extracts were basified with aqueous sodium hydroxide and the liberated phosphine extracted into chloroform. The solvent was removed from the combined chloroform extracts under reduced pressure to give the required phosphine (0.4 g). δ³¹P (CDCl₃) _ 63.1 δ¹³C (CDCl₃) 15.5(t, J = 5), 15.9(t, J = 5), 38.1(dd, J = 16, 11), 42.2(t, J = 10), 58.6(s), 58.8(s), 70.3(s), 71.6(s), 74.0(t, J = 9), 76.5(t, J = 9). EXAMPLE 12 Preparation of 4,4-Bis(dimethylphosphinomethyl) tetrahydropyran 1. 4,4-Bis(ethoxycarbonyl)tetrahydropyran Sodium (10 g) was dissolved in dry ethanol (300 cm³) and then diethyl malonate (30 g) was added slowly. After stirring for 15 minutes, bis(2-chloroethyl)ether (35 g) was added dropwise. The mixture was heated under reflux for 72 hours and then cooled to room temperature. After filtering the solvent was removed from the reaction mixture under reduced pressure to give the crude product. An initial purification was achieved by chromatography on alumina (activity 4) using ethyl acetate/petroleum ether mixtures as eluants. The pure product (11.3 g) was isolated by distillation under reduced pressure as an oil (b.p. 106-108°C at 0.03 mmHg). δ¹³C (CDCl₃) 13.8(×2), 30.8(×2), 52.1, 61.3(×2), 64.5(×2), 170.6(×2). ii. 4,4-Bis(hydroxymethyl)tetrahydropyran 4,4-Bis(ethoxycarbonyl)tetrahydropyran (11 g) in dry diethyl ether (50 cm³) was added slowly to a stirred suspension of lithium aluminium hydride (2 g) in dry diethyl ether (15 cm³) at a rate so as to maintain gentle boiling. Following the addition, the mixture was heated under reflux for a further 1 hour and then allowed to cool. Ethyl acetate (4 cm³) was slowly added, followed by water (1.8 cm³), then aqueous sodium hydroxide (1.8 cm³), 15%, and finally a second portion of water (5 cm³). The mixture was filtered and the solid washed with diethyl ether. The combined ether extracts were dried over anhydrous sodium sulphate and the ether then removed to give the diol (5.5 g). δ¹³C (CDCl₃) 29.1(×2), 36.0, 63.3(×2), 67.4(×2) iii. 4,4-Bis(chloromethyl)tetrahydropyran 4,4-Bis(hydroxymethyl)tetrahydropyran (5.5 g), triphenylphosphine (19.8 g), dry carbon tetrachloride (100 cm³) and chloroform (15 cm³) were boiled together under reflux in an atmosphere of dry nitrogen until the reaction was complete (about 8 hours, as shown by nmr spectroscopy). The volatile components were removed under reduced pressure and the resulting solid mass extracted with petroleum ether (b.p. 40-60°C). Evaporation of the petrol from these extracts gave the crude dichloride. A sample of the pure dihalide (1.2 g) was obtained by vacuum distillation (b.p. 60°C at 0.05 mmHg) followed by chromatography on Kieselgel 60 (Merck) using ethyl acetate/petroleum ether mixtures as eluants. δ¹³C (CDCl₃) 31.4(×2), 37.2, 48.4(×2), 63.0(×2) iv. 4,4-Bis(dimethylphosphinomethyl)tetrahydropyran To a stirred solution of dimethylphosphine (5-cm³) in dry liquid ammonia (60 cm³) was added a solution of n-butyl lithium (10.5 cm³, 2.4 M in hexanes). After 10 minutes at 60°C 4,4-bis(chloromethyl)tetrahydropyran (1 g), in a little diethyl ether, was added. After a further period of 15 minutes at low temperatures the mixture was allowed to warm up slowly to room temperature. When the ammonia had evaporated diethyl ether (150 cm³) was added and the resulting mixture extracted with dilute hydrochloric acid (2 M). The combined aqueous extracts were basified with aqueous sodium hydroxide and the liberated phosphine extracted into chloroform. The solvent was removed from the combined chloroform extracts under reduced pressure to give the phosphine (0.3 g). δ³¹P (CDCl₃) _ 62.5 δ¹³C (CDCl₃) 15.7(×2)(t, J = 4), 16.1(×2)(t, J = 4), 34.7(t, J = 12), 37.8(×2)(t, J = 8), 43.4(×2)(dd, J = 16, 12), 63.5(×2)(s). EXAMPLE 13 By routes corresponding to those described in Examples 8 to 12, the following ligands were prepared. EXAMPLE 14 [Tc(I)L₃]⁺ Technetium (I) trisdiphosphine complex L = Ligand of Example 8 Materials 10 mg Na₂S₂O₄ 1 ml Saline 2 ml EtOH 0.05 ml 10 M NaOH 2O µl L 1 ml 99 m TcO4 _ Na generator eluate (at 2.9 GBq/ml) Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 30 minutes. After cooling, the pH was adjusted to 6.5 with 0.1 M HCl and diluted with 4 ml of saline. The resulting solution was then submitted to chromatography analysis and animal biodistribution studies. Chromatographic Data The resulting preparation contains no free TcO₄ _ or reduced technetium colloid, and indicates that the desired species is present in solution approximately 85% yield. Saline rf = 0.09 Methylethylketone rf = 0.60 (broad) Acetonitrile = Water 50: 50 rf = 0.90 HPLC Data The complex elutes as a sharp band at approximately 18.3 minutes (∼ 15% minor component at ∼ 17.0 minutes). Biodistribution Results See Tables IX and X. EXAMPLE 15 Synthesis of [Tc(NO)X(L)₂]⁺X _ Nitrosyl complex X = Cl L = Ligand of Example 8 Materials NH₂OH · HCl = 25 mg SnF₂ = 1.0 ml at 6.6 × 10 _5 M solution (aq). L = 10 µl 99 m TcO₄ _ Na, 0.7 ml, generator eluate at 3.58 GBq/ml Saline 0.3 ml Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 1 hour. After cooling, the prep was filtered through a filter (an acrodisc 0.2 µm). The resulting solution was then submitted to chromatographic analysis and animal biodistribution study. Chromatography The resulting prepartion contain no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 90% yield. Saline rf = 0.04 Methylethylketone rf = 0.62 Acetonitrile: Water 50: 50 rf = 0.98 HPLC Data The complex elutes as a sharp peak at approximately 17.4 minutes. Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.89 (_ indicating movement towards cathode). Biodistribution Results See Table XI EXAMPLE 16 Synthesis of [Tc(L)₃]⁺ technetium (I) trisdiphosphine complex. L = Ligand of Example 10. Materials 2 ml EtOH 1 ml SnF₂ 2.64 × 10⁴ M solution (aq). 10 µl L 2 ml TcO₄ _ /Saline 99 m TcO₄ _ Na generator eulate at 0.72 GBq/ml. Methods The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 1 hour. After cooling, the preparation was submitted for animal biodistribution studies. Chromatography The resulting preparation contains no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 70% yield. Saline rf = 0.02 Methylethylketone rf = 0.69 (70%) 0.03 (30%) Acetonitrile: Water 50: 50 rf = 0.98 HPLC Data The complete elutes as a single band with a retention of approximately 19.8 minutes (two minor components at 14 minutes and 18 minutes). Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.53 (with two minor components with rf = + 0.04 and _ 1.07, where _ indicates movement towards cathode). Biodistribution Results See Tables XII and XIII EXAMPLE 17 [Tc¹NO(X)(L)₂]⁺Cl _ X = Cl L = Ligand of Example 10 Materials NH₂OH · HCl 25 mg SnF₂ 1 ml. of 6.6 × 10 _5 M solution. Saline 2.0 ml L 10 µl 99 m TcO₄ _ Na⁺ 2.0 ml Generator eluate (6.1 GBq/ml) Methods The components were mixed in a sealed nitrogen purged vial, and heated at 120°C for 1 hour. The cooled prepartion was filtered through 0.2 µm acrosdisc filter (Gelman) and diluted with 8 ml of saline. The resulting solution was submitted for chromatography analysis and animal biodistribution studies. Chromatography The resulting preparation contains no colloid and free TcO₄ _ , and indicates that the desired species is present is approximately 80% yield. Saline rf = 0.06 Methylethylketone rf = 0.74 (80%) 0.01 (20%) Acetonitrile: water 50: 50 rf = 0.99 Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.78, (_ indicates movement towards cathode) Biodistribution Results See Tables XIV and XV EXAMPLE 18 Synthesis of [Tc(I)(L)₃]⁺ Technetium trisdiphosphine complex L = Ligand of Example 9 Materials EtOH 0.5 ml Saline 3 ml L 10 µl 99 m TcO₄Na 0.3 ml Generator eluate at 4.17 GBq/ml. Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 1 hour. After cooling the preparation was submitted for chromatography analysis and biodistribution studies. Chromatography The resulting preparation contains no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 90% yield. Saline rf = 0.06 Methylethylketone rf = 0.72 Acetonitrile: Water 50: 50 rf = 0.99 HPLC Data The complex elutes as a broad band with a retention time of approximately 7.1 minutes. Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.27 (_ indicates movement towards cathode). Biodistribution results See Tables XVI and XVII EXAMPLE 19 Synthesis of [Tc I (NC)X(L)₂]⁺Cl Technetium Nitrosyl diphosphine complex L = Ligand of Example 9 Materials NH₂OH · HCL 25 mg Saline 3.7 ml L 10 µl 99 m TcO₄Na 0.3 ml Generator eluate at 8.75 GBq/ml. Method The components were mixed in a sealed, N₂ purged vial and heated at 120°C for 1 hour. After cooling the preparation was filtered through 0.2 µm acrodisc (Gelman). The resulting preparation was then submitted for chromatography analysis and biodistribution studies. Chromatography The resulting preparation contains no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 90% yield. Saline rf = 0.01 Methylethylketone rf = 0.78 Acetonitrile: Water 50: 50 rf = 0.99 HPLC Data The complex elutes as a single band with a retention time of approximately 6.7 minutes (a minor component is at approximatly 5.8 mintues) Gel Electrophoresis The complex moves as a single band towards the cathode rf = _ 0.64 (_ indicates movement towards cathode). Biodistribution results See Tables XVIII and XIX EXAMPLE 20 Synthesis of [Tc(I)(L)₃]⁺X _ Technetium (I) trisdiphosphine complex L = Ligand of Example 11. Materials EtOH 0.5 ml Saline 3 ml L 15 µl 99 m TcO₄Na 0.35 ml Generator eluate at 4.26 GBq/ml. Method The components were mixed in a sealed, N₂ purged vial and heated at 60°C for 1 hour. After cooling the resulting preparation was submitted for chromatography analysis and biodistribution studies. Chromatography The resulting preparation contains no colloid or free TcO₄ _ , and indicates that the desired species is present in approximately 90% yield. Saline rf = 0.00 Methylethylketone rf = 0.71 Acetonitrile: Water 50: 50 rf = 0.01 HPLC Data The complex elutes as a single peak with a retention time of approximately 6.3 minutes. Gel Electrophoresis The complex moves eluates as a single band towards the cathode rf = _ 0.60 (_ indicates movement towards cathode). Biodistribution results See Tables XX and XXI EXAMPLE 21 Preparation of P46 : (MeOC₂H₄OCH₂)MePC₂H₄PMe(CH₂OC₂H₄OMe) Reaction Scheme : Experimental : All reactions and manipulations were performed under vacuo or oxygen-free nitrogen atmosphere. Solvents were dried, and degassed by nitrogen purge prior to use. CH₃OCH₂CH₂OCH₂Cl and n-BuLi were purchased from Aldrich. Me(H)PC₂H₄P(H)Me was prepared according to a published method (1) . 1. M Baake, O Stelzer, and V Wray. Chem. Ber. 113 , 1356 (1980). Procedure : In a 250 cm³, 3-necked round bottomed flask, equipped with a dry ice condenser, pressure equallising dropping funnel and a teflon stirring bar, was placed in a solution of Me(H)PC₂H₄P(H)Me (3.18 g, 26.06 mmol) in petrol (40-60°C, 50 cm³). To this solution was added n-BuLi (35 ml, 57.0 mmol, 1.6 M in hexane) at _78°C. After warming back to room temperature a white precipitate was filtered and washed with petrol (40-60°C, 2 × 25 cm³). Sodium-dried liquid ammonia (150 cm³) was then condensed into the above reaction vessel and an orange solution was formed. To this a solution of CH₃OC₂H₄OCH₂Cl (6 cm³, 52.1 mmol) in diethyl ether (20 cm³) was added dropwise until the orange colour just disappeared. Then the ammonia was allowed to vaporise at ambient temperature. Diethyl ether (30 cm³) was added to give a white suspension. This subsequently was hydrolysed, and the organic layer was separated and dried over anhydrous magensium sulphate. Diethyl ether was removed by distilation at atmospheric pressure. The rest of the volatile materials were removed by distillation under vacuo at 120°C. The colourless liquid left was pure by NMR. Yield = 4.7 g, 50.3%. EXAMPLE 22 [Tc(P46)₃]⁺ Materials 1.5 ml SnF₂ aqueous solution 32 µg per ml 2 ml Ethanol 0.05 ml 10 M NaOH 10 µl P46 2 ml Tc 99 m O₄ (_) Na (+) generator eluate at 1.78 GBq per ml Method : The components were mixed in a sealed, N₂ purged vial and left standing at room temperature for 2 hours. Then pH was adjusted to 7 with diluted HCl acid (1 M). The resulting solution was then submitted to chromatographic analysis and animal biodistribution study. Chromatography Data : The resulting solution (above) contains no colloid or free Tc 99m O₄ _ and indicates that the Tc complex present in solution is approximately 70% pure. Saline rf = 50.00 Methyl Ethyl Ketone rf = 50.00 (30%) rf = + 0.62 (70%) Acetonitrile/water 50: 50 rf = + 0.98 HPLC Data : The complex elutes as a sharp peak at approximately 6.7 minutes plus peaks at 5.5 and 6 minutes. Gel Electrophoresis Data : The complex moved as a single band towards the cathode rf = _ 0.34 (_ indicating movement towards cathode. Biodistribution Results : See Tables XXII and XXIII Experimental and Chromatography, Electrophoresis, HPLC and in vivo biodistribution are exactly the same as previously described. Example 23 Materials Method : Firstly, the FeCl₃.6H₂O and P46 in ethanol were mixed in a P6 vial. A purple solution was formed instantaneously. Then this purple solution was transferred into a P11 vial which contained NaCl, EGTA, saline and 99 m TcO₄ _ . The reaction mixture was then heated at 120°C for 60 minutes (pH = 4). The resulting solution was submitted to various analytical techniques, summarised as below: Chromatography Data : The resulting solutions (above) contain no colloid or free 99 m TcO₄ _ and indicates that the technetium complex is present in solution in approximately 80% yield. Saline rf = 0.01 Methyl Ethyl Ketone rf = 0.59, 0.02 Acetonitrile/water rf = 0.99 HPLC Data : The complex elutes as a sharp peak at approximately 7.3 minutes, plus two small peaks at 5.2′ and 6.1′. Gel Electrophoresis Data : The complex moved as a single band towards the cathode rf = _ 0.67 (_ indicating movement towards cathode). Biodistribution Results : See Table XXIV EXAMPLE 24 By methods generally as described in Examples 14 to 20, ligands listed in Example 13 were used to prepare cationic Technetium-99 m complexes which were subjected to biodistribution testing in rats. The results are summarized in the following Tables XXV and XXVI. EXPERIMENTAL The experimental techniques used to characterise and evaluate these new radiopharmaceutical complexes are outlined below: Chromatography Samples were supplied by needle approximately 2.5 cm from the bottom of two Gelman ITLC/SG strips (2.5 cm × 20 cm) and one Whatman No.1 strips (2.5 cm × 20 cm) and then immediately placed in prepared ascending chromatography development tanks containing fresh solvent (1 cm): a) saline, b) methyl ethyl ketone, and c) 50: 50 acetonitrile: water respectively. After 15 cm elution the strips are removed, solvent fronts marked, dried and the distribution of activity determined using suitable equipment. Electrophoresis An 0.1 g agarose/10 cm³ phosphate buffer pH 7.4 gel was run at an applied potential of 300 V for approximately 35 mins, using bromophenol blue indicator (this indicator moves towards the cathode). The resulting distribution of activity was determined using suitable equipment. HPLC For Examples 3 to 7 and 18 to 23, a solvent gradient HPLC system was used, in conjunction with: a) 20 mM phosphate buffer pH 7.4 b) Tetrahydrofuran (THF) Samples are applied initially at 100% (a), the gradient changing to 100% (b) in approximately 17 minutes. Flow rate = 2 ml/min Hamilton PRP column (15 cm × 4.0 mm) ambient temperature. Identical HPLC systems were used for 99 m Tc and ⁹⁹Tc determinations, 99 m Tc detected by emission, ⁹⁹Tc detected via liquid scintillation method. For Examples 14 to 17 the system was the same except solvent gradient system (a) 100 nM sodium acetate pH 5.5 (b) Acetone Samples are applied at 100% (a), which is held for 5 minutes and then gradient changing to 100% (b) in 20 minutes. Held at 100% (b) for further 5 minutes. Flow rate = 2 ml/min Hamilton PRP column. ANIMAL BIODISTRIBUTION In vivo studies In vivo biodistribution: 0.1 ml Tc 99 m prep was injected i.v. into a laterial tail vein of 6 anaesthetised rats. At 2 minutes and 60 minutes post-injection, 3 rats were sacrificed by decapitation, bled from the neck and dissected. The following organs were removed at dissection: kidney; bladder (+ urine), lung, liver, spleen, stomach, small intestine, large intestine, brain (weighed), heart (weighed), thyroid and samples of blood (weighed) and muscle (weighed), the residual carcass and the tail (injection site). Subsequently samples were counted in an automatic twin crystal gamma counter. Percentage biodistribution of injected material was calculated (after correction for background) for all organs using the formula: Since only samples of muscle and blood were taken, the percentage in these tissues was calculated assuming blood and muscle to represent 5.8 and 43% of total animal weight respectively using the formual: Since only samples of muscle and blood were taken, the percentage in these tissues was calculated assuming blood and muscle to represent 5.8 and 43% of total animal weight respectively using the formula: where CF = 0.058 for blood 0.43 for blood References 1. Deutsch, E., Libson, K., Jurisson, S., Lindoy, L.F., Technetium Chemistry and Technetium Radiopharmaceuticals Prog. Inorg. Chem. 1982, Vol.30, p.175

Claims: 1. A cationic complex of technetium-99 m with a ligand having the formula: Y₂PZPY₂ where the groups Y may be the same or different and each is H or C1-C8 saturated hydrocarbon or saturated fluorohydrocarbon which may contain up to 3 ether oxygen atoms, Z is a _CC_ or _CCC_ or _COC_ chain which may be substituted by at least one C1-C8 saturated hydrocarbon or saturated fluorohydrocarbon group which may contain up to 3 ether oxygen atoms, provided that the ligand contains at least one _COC_ ether linkage. 2. A cationic complex as claimed in claim 1, wherein the groups Y may be the same or different and each is H or C1-C4 alkyl which may be sustituted by C1-C4 alkoxy, Z is a _CC_ of _CCC_ or _COC_ chain which may be substituted by C1-C4 alkoxy or alkoxyalkyl or spirocyclic ether. 3. A cationic complex as claimed in claim 1, wherein Z is a _CC_ or _CCC_ chain, each Y is H or methyl, and either one or more methoxy substituents is attached to one or more groups Y, or one or more methoxymethyl or _COC_ spirocyclic ether groups is attached to a carbon atom of Z. 4. A cationic complex as claimed in claim 1 wherein the ligand has the formula: 5. A cationic complex as claimed in claim 1 wherein the ligand has the formula: 6. A cationic complex as claimed in claim 1 wherein the ligand has the formula: 7. A cationic complex as claimed in claim 1 wherein the ligand has the formula: 8. A cationic complex as claimed in any one of claims 1 to 7, having a formula selected from [Tc(NO) n X m L₂]⁺A _ and [TcL₃]⁺A _ where X is a monodentate ligand for Tc A is an anion n is 1 or 2 and m is correspondingly 1 or 0, and L is a ligand as claimed in any one of claim 1-7. 9. A cationic complex as claimed in claim 8, having the formula: Tc(NO) X L₂]⁺A _ where L is a ligand having the formula 10. A cationic complex as claimed in claim 8, having the formula [TcL₃]⁺A _ where L is a ligand having the formula 11. A cationic complex as claimed in any one of claims 1 to 10, for use as a heart imaging agent. 12. A ligand having the formula Y₂PZPY₂, where the groups Y may be the same or different and each is C1-C4 saturated hydrocarbon, and Z is a _CCC_ or _COC_ chain which may be substituted by at least one C1-C8 saturated hydrocarbon or saturated fluorohydrocarbon groups which may contain up to 3 ether oxygen atoms, provided that the ligand contains at least one _COC_ ether linkage. 13. A ligand as claimed in claim 12, wherein Z is a _CCC_ or _COC_ chain which is substituted by at least one C1-C4 alkoxy or alkoxyalkyl or spirocyclic ether group. 14. A ligand as claimed in Claim 12, wherein Z is a _CCC_ chain with one or more methoxymethyl or _COC_ spirocyclic ether groups attached to a carbon atom of Z. 15. A ligand as claimed in Claim 12 having the formula 16. A ligand as claimed in Claim 12 having the formula

Inventors: CHIU, KOWK WAI, EDWARDS, PETER GERALD, GRIFFITHS, DAVID VAUGHAN, KELLY, JAMES DUNCAN, LATHAM, IAN ANDREW
Applicants: AMERSHAM INTERNATIONAL PLC
Application year: 1988
Main classification: C07F9
IPC codes: A61K51, C07F9, C07F13