Aza macrocycles and processes for their preparation

Abstract: Aza macrocycles of the formula (1) wherein A is a group -(CH₂) p - or -(CH₂) p N(R)(CH₂) q - where R is a group -CH(R⁴)X⁴; m, n, p, and q, which may be the same or different, is each an integer 2 or 3; R¹, R², R³ and R⁴, which may be the same or different, is each a group -CO₂H, -P(R⁵)O₂H (where R⁵ is a hydrogen atom or an alkyl or alkoxy group), -PO₃H₂ or -CONR⁶R⁷ (where R⁶ and R⁷, which may be the same or different is each a hydrogen atom or an alkyl group); X¹, X², X³ and X⁴, which may be the same or different, is each a hydrogen atom or an alkyl group, or a linker group, with the proviso that at least one of X¹, X², X³ or X⁴ is a linker group; and protected derivatives and metal complexes and/or salts thereof; are described together with processes for their preparation and compositions containing them. The compounds are useful for imaging and in the treatment of abnormal cell disorders, such as in the treatment of tumours, and may be coupled to other molecules such as proteins for use in diagnosis and therapy.

Description: Field of the Invention This invention relates to functionalised aza macrocycles, to metal complexes thereof, to conjugate compounds containing the functionalised aza macrocycles and metal complexes thereof and to their use in diagnosis and therapy. Background to the Invention The attachment of metal ions to proteins, peptides and other, smaller molecules is a fast expanding technology, which has numerous proven and potential applications in research, in industry and, particularly, in medicine. In recent years, much of the impetus behind the development of this technology has been the ability to link metal ions to antibodies, especially monoclonal antibodies. Such metal labelled antibodies have found a widespread use, especially in medicine, where they have been employed, for example, to target the metal ion to a specific tissue type, both in vitro and in vivo . Thus, metal labelled antibodies have applications in locating specific tissue types (e.g. employing computer-aided tomographic techniques where the metal ion is in some way detectable) and in the treatment of cell disorders (e.g. treating mammalian tumours where the metal ion is a cytotoxic radionuclide). Conventionally, attachment of the metal ion to a protein such as an antibody has been achieved by complexation by an acyclic chelate such as a substituted diethylenetriaminepentaacetic acid [Gansow O. A. et al , Inorg. Chem., (1986), 25 , 2772] or ethylenediaminetetraacetic acid [Meares, C. F. et al , Acc. Chem. Res., (1984), 17 , 202] covalently linked to the antibody. Such acyclic complexes however tend to be unstable in vivo either as a result of acid-catalysed decomplexation or competitive chelate binding by Ca²⁺ or Zn²⁺ in serum, or as a result of competition from transferrin [Moerlein, S. M. et al , Int. J. Nuc. Med. Biol., (1981) 8 , 277]. The lack of stability can result in uncomplexed metal atoms in the body which have a cytotoxic effect on healthy tissue (e.g. bone marrow) or which markedly reduce the signal-to-noise ratio of an imaging technique. A possible alternative to the use of acyclic chelates in the labelling of antibodies is the use of macrocyclic ligands, which has been suggested by a number of workers [Gansow O. A. et al . Am. Chem. Soc. Symp. Ser., (1984), 241 , 215; UK Patent Specification Publication No. 2122641; International Patent Specifications Nos. WO89/01475 and WO89/01476 and European Patent Specification No. 305320; and Moi M. K. et al , Anal. Biochem., (1985), 148 , 249-253]. We have now found a new class of functionalised aza macrocycles, members of which are able to form kinetically inert complexes with metal ions. The macrocycles of the invention are particularly useful for attachment to proteins, especially antibodies, to provide conjugate compounds capable of binding metals to give complexes which are advantageously stable in vivo . Thus, according to one aspect of the present invention we provide a compound of general formula (1); wherein A is a group -(CH₂) p - or -(CH₂) p N(R)(CH₂) q - where R is a group -CH(R⁴)X⁴; m, n, p, and q is each an integer 2; R¹, R², R³ and R⁴, which may be the same or different, is each a group -CO₂H, -P(R⁵)O₂H (where R⁵ is a hydrogen atom or a C₁₋₆ alkyl or alkoxy group), -PO₃H₂ or -CONR⁶R⁷ (where R⁶ and R⁷, which may be the same or different is each a hydrogen atom or a C₁₋₆ alkyl group); X¹, X², X³ and X⁴, which may be the same or different, is each a hydrogen atom or a C₁₋₆ alkyl group, or a linker group of the formula -L-Z wherein L and Z are as defined below, with the proviso that at least one of X¹, X², X³ or X⁴ is a linker group; and metal complexes and/or salts thereof. It will be appreciated that formula (1) [and, where appropriate, the following formulae herein], is intended to cover all stereoisomers of the compounds concerned, including mixtures thereof. In the compounds of formula (1), it will be appreciated that the nature of the linker group may be varied widely without substantially affecting the usefulness of the compounds. In the following definition, and in the same context whenever it appears below, the term "interrupted by" as applied to cycloaliphatic or aromatic groups is to be understood to also mean that these particular groups may additionally be present linked to the terminal carbon atom of the hydrocarbyl chain represented by L, at the opposite end of the chain to the carbon atom attached to the macrocycle. L is an optionally substituted straight or branched C₁₋₂₀alkylene, C₂₋₂₀alkenylene, or C₂₋₂₀alkynylene chain, optionally interrupted by one or more -O- or -S- atoms or C₅₋₈cycloalkylene (e.g. cyclopentylene or cyclohexylene), C₆₋₁₂aromatic (e.g. phenylene or substituted phenylene), C₅₋₁₀heteroaromatic (e.g. furanyl, pyridyl), -N(R⁸)-, -CON(R⁸)- or -N(R⁸)CO- groups (where R⁸ is a hydrogen atom or a C₁₋₆ alkyl group). Examples of substituents which may be present on the chain L include halogen atoms, e.g. fluorine, chlorine, bromine, or iodine atoms or groups selected from C₁₋₆alkoxy (e.g. methoxy or ethoxy), hydroxy, nitro, -N(R⁹)(R¹⁰), [where R⁹ is a hydrogen atom or a C₁₋₆alkyl group and R¹⁰ is a C₁₋₆alkyl group; e.g. -NHCH₃ or -N(CH₃)₂], or substituted amido, e.g. a group of formula -(CH₂) d CON(R¹¹)(R¹²) [where d is zero or an integer 1 to 4 inclusive, R¹¹ is a hydrogen atom or a C₁₋₆alkyl group, e.g. methyl and R¹² is an optionally substituted C₁₋₆alkyl group]. Substituted alkyl groups represented by R¹¹ include for example C₁₋₆alkyl groups substituted by one or more halogen atoms, or nitro, amino or hydroxy groups. Z is a halogen atom, for example a chlorine, bromine or iodine atom, or a group selected from OH, -SH, -NH₂, hydrazine (-NHNH₂), or a derivative thereof, [for example -N(CH₃)NH₂, -NHCONHNH₂, -NHCSNHNH₂, or phenyl hydrazine], -NCO, -NCS, -COR¹³, [where R¹³ is a halogen atom such as a chlorine or bromine atom, or a N₃, C₁₋₆alkoxy, e.g. methoxy, C₆₋₁₂aryloxy (e.g. nitrophenyloxy or dinitrophenyloxy), imidyloxy (e.g. succinimidyloxy) or imidazolyloxy group], imide,e.g. maleimide, a vinyl group of formula -Het¹-C(Het²)=CH₂ (where Het¹ and Het², which may be the same or different, is each a nitrogen containing heterocyclic group, e.g. a pyridyl group or Het¹ is a nitrogen containing heterocyclic group and Het² is a hydrogen atom), for example a vinyl pyridyl group of formula (where R¹⁴ is a C₁₋₄alkyl e.g. methyl group). Alkyl groups represented by the groups X¹, X², X³, X⁴, R⁵, R⁶ or R⁷ include C₁₋₆alkyl groups such as methyl or ethyl groups. Metal complexes of the compounds of formula (1) in which A is -(CH₂) p N(R)(CH₂) q - include complexes wherein the metal is di- or tripositive and has a coordination number 6 or greater, especially 8. Examples of such metals include indium (In), copper (Cu), lead (Pb), bismuth (Bi), yttrium (Y), terbium (Tb), gallium (Ga), gadolinium (Gd) and scandium (Sc). In, Y, Ga, Tb, Gd, and Sc are preferred, particularly In, Y, Gd, and Ga. In general the metal is preferably a radioactive isotope. Yttrium, especially ⁹⁰Y, is particularly preferred. Metal complexes of the compounds of formula (1) in which A is -(CH₂) p - include complexes wherein the metal is di- or tri-positive and has a coordination number from 2 up to 6, especially 6. Examples of such metal(s) include indium (In), copper (Cu), lead (Pb), bismuth (Bi), cobalt (Co) and gallium (Ga). In, Ga, Co and Cu are preferred, particularly In and Ga. In general the metal is preferably a radioactive isotope. Indium, especially ¹¹¹In, is particularly preferred. In general, optimum binding of the metal to the compounds of formula (1) may be achieved by selection of the ring size and where appropriate by adjusting the potential coordination number by choice of the group R¹, R², R³ or R⁴. Salts of the compounds of formula (1) include salts with bases, e.g. sodium or potassium salts, or acid addition salts such as hydrobromides or hydrochlorides. Pharmaceutically acceptable salts are particularly preferred. A preferred group of compounds of formula (1) wherein A is -(CH₂) p - or -(CH₂) p N(R)(CH₂) q - is that wherein R¹, R², R³, and R⁴ is each a -CO₂H group. A further preferred group of compounds of formula (1) wherein A is -(CH₂) p - or -(CH₂) p N(R)(CH₂) q - is that wherein one or two of X¹, X², X³ and X⁴ is a linker group and the remainder are alkyl groups or, especially, hydrogen atoms. One group of compounds of formula (1) has the formula (1a): wherein m, n, p, R¹, R², R³, X¹, X² and X³ are as defined for formula (1); and metal complexes and/or salts thereof. Indium complexes of the compounds of formula (1a) are particularly preferred. Particularly important compounds of formula (1a) are those of formula (1b) wherein one or two of X¹, X², or X³ is a linker group and the remainder are hydrogen atoms; and metal complexes and/or salts thereof. Indium complexes of the compounds of formula (1b) are particularly preferred. Another group of compounds of formula (1) has the formula (1c): wherein m, n, p, q, R¹, R², R³, R⁴, X¹, X², X³ and X⁴ are as defined for formula (1); and metal complexes and/or salts thereof. Yttrium complexes of the compounds of formula (1c) are particularly preferred. An important group of compounds of formula (1c) are those of formula (1d) wherein one or two of X¹, X², X³ or X⁴ is a linker group and the remainder are hydrogen atoms; and metal complexes and/or salts thereof. Yttrium complexes of the compounds of formula (1d) are particularly preferred. The compounds of formula (1) and the metal complexes and/or salts thereof have a diagnostic use as imaging agents in vitro and in vivo . The compounds of formula (1) and the metal complexes and/or salts thereof are also cytotoxic agents and may be used in the treatment of abnormal cell disorders, for example in the treatment of tumours. For use as diagnostic and/or therapeutic agents, the compounds of formula (1) may be employed using conventional methods, (e.g. for formulation and presentation) already in use for metal complexing agents. For application of the compounds of formula (1) as imaging or cytotoxic agents, it is generally preferable to couple the compounds to other molecules such a proteins, especially antibodies, peptides or carbohydrates to form conjugate compounds, and the compounds of formula (1) are particularly well adapted for use in this respet. Thus, according to a further aspect of the invention, we provide a conjugate compound which comprises a compound of formula (1), or a metal complex and/or salt thereof, coupled to a protein, peptide or carbohydrate. The compound of formula (1) may be coupled through any thiol, amino, carboxyl, hydroxyl, aldehyde, aromatic or heteroaromatic group present in the protein, peptide or carbohydrate. In a preferred aspect of the invention, we provide a conjugate compound which comprises a compound of formula (1) or a metal complex and/or salt thereof, coupled to an antibody. It is to be understood that conjugate compounds according to the invention may contain more than one molecule of a compound of formula (1) couple to any one protein, peptide or carbohydrate molecule. Particularly useful conjugate compounds according to the invention are those comprising a compound of formula (1b) or formula (1d) or a metal complex and/or salt thereof, coupled to an antibody. The indium and yttrium complexes of these conjugates are especially important. The compounds of formula (1) and conjugate compounds of the invention may be formulated for use in accordance with conventional practice. Thus according to a further aspect of the invention we provide a composition comprising a compound of formula (1); or a conjugate compound comprising a compound of formula (1) coupled to a protein, peptide or carbohydrate, or a metal complex and/or salt thereof, together with one or more pharmaceutically acceptable carriers. Particularly suitable compositions according to the invention are those adapted for parenteral administration, especially by injection or infusion. Suitable formulations of this type include suspensions solutions or emulsions of the compound or conjugate in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilising and/or dispersing agents. Alternatively the compound or conjugate may be in powder form for reconstitution with a suitable vehicle, e.g. sterile pyrogen-free water before use. If desired the compound or conjugate may be presented in unit dosage form, and/or together with one or more active ingredients or imaging agents. Suitable formulations of this type include solutions of the compounds of formula (1) in isotonic saline. The quantities of compounds of formula (1) used in formulations according to the invention will vary according to the intended cell target, but may be easily determined in accordance with conventional practice for reagents of this type. Compounds of the invention may be prepared by the following processes wherein the groups and symbols R¹, R², R³, R⁴, m, n, p, q, X¹, X², X³ and X⁴ are as defined for formula (1) except where stated otherwise. Where a metal complex is desired as a final product, the complexation with a metal atom may be carried out as a final step in the production process, as described below for the complexation of compounds of formulae (1), or alternatively it may be desirable to complex the metal at an earlier stage in the process, providing of course that the requisite macrocycle structure is present. In the following processes, it may be desirable to use starting materials in which functional groups in the linker group are in a protected state, or which contain a precursor of the group, as discussed below. Thus according to a further aspect of the invention a compound of formula (1) wherein at least one of X¹, X², X³ and X⁴ is other than a linker group, or a metal salt thereof may be prepared by reaction of a compound of formula (2): [wherein A¹ is -(CH₂) p - or -(CH₂) p N(H) (CH₂) q - a least one of the groups X is a hydrogen atom and the remainder is each a group -CH(R¹)X¹, where R¹ is as previously defined and X¹ is a linker group] with a compound DCH(R²)X² (where X² is as defined for formula (1), but is not a linker group, and D is a displaceable group, for example a halogen atom such as a bromine atom) followed, where necessary, by removal of any protecting group. The reaction may be performed in a solvent such as water or an organic solvent such as a nitrile, e.g. acetonitrile, or an alcohol, e.g. ethanol, or an amide, e.g. dimethylformamide, in the presence of a base such as an alkali metal carbonate or hydroxide, e.g. sodium, potassium or caesium carbonate, or sodium, potassium or lithium hydroxide, at an elevated temperature e.g. the reflux temperature. Where metal complexes of formula (1) are required (or any other suitable macrocylic intermediate described herein) these may be prepared by treating the compound with a metal salt (for example a metal halide) in an appropriate solvent for example an aqueous or non aqueous solvent, (e.g. acetonitrile, acetone, propylene carbonate, dimethylformamide or dimethylsulphoxide) at any suitable temperature from 0°C to 100°C such as 10°C to 80°C e.g. around 60°C. A conjugate compound according to the invention may be prepared by reaction of a compound of formula (1) or a metal complex thereof with a protein, peptide or carbohydrate in a aqueous solvent, for example an inorganic buffer such as a phosphate buffer at an appropriate temperature. Salts of compounds of formula (1) and their metal complexes may be prepare by conventional means, for example by reaction with an appropriate base or acid in a suitable aqueous solvent. Intermediates of formula (2) may be prepared by reaction of a compound of formula (3) with a compound DCH(R¹)X¹ (where X¹ is a linker group or a protected derivative thereof) in the presence of a base in a suitable solvent at an elevated temperature as just described for the preparation of compounds of formula (1). By varying the molar ratio of the compound of formula (2) and the compound DCH(R¹)X¹ such that the latter is increased relative to the former, (for example from around 2:1 to 1:1 and further) compounds of formula (2) containing more than -CH(R¹)X¹ group as just defined may be prepared. Intermediates of formula (3) are either known compounds or may be prepared from known starting materials using methods analogous to those used for the preparation of the known compounds. The following Examples illustrate the invention. Example 1 (a) To a stirred solution of 1,4,7,10-tetraazacyclododecane (0.057g) in dry dimethylformamide (5ml) was added anhydrous potassium carbonate (0.05g). To this mixture at 80°C under nitrogen was added 2-bromo-6-benzamidoethyl hexanoate (0.113g) in dimethylformamide (5ml) over 2 hours. HPLC analysis revealed that after 2 days reactions was essentially complete to give, after removal of dimethylformamide a single major product [CM-300; gradient elution; t=0: A=80%, B=0%, C=20%; t=5min: A=60%, B=20%, C=20%; t=10 min: A=0%, B=80%, C=20% (with A=H₂O, B=1M ammonium acetate (pH5.6), C=CH₃CN) flow rate = 1.4ml:min⁻¹]. Retention time = 7.5min. m/e (CI) 433 (M⁺+1) δ H (CDCl₃) 1.25 (3H, t, CO₂CH₂ CH ₃), 1.45-1.79 (6H, mult, CH 2 CH 2 CH 2 CH 2 NH), 2.39-3.13 (16H m.ring CH₂'s) 3.32-3.50 (3H, m, CONH C H₂ and OCOC H ), 4.14 (2H, q, CH₂O), 7.37-7.50 (4H, m, arom plus NHCO), 7.79 (2H, d+d ortho arom H). (b) The compound prepared in part (a) was not isolated but was redissolved in dimethylformamide (5ml). To this solution was added dry potassium carbonate (0.15g) and at 80°C ethyl bromoacetate (0.165g) was added to a solution in dimethylformamide (2ml) over 15 min. Reaction was complete in 4 hours [HPLC analysis gave a major product at t=6.19 min under the conditions defined in part (a)]. After removal of dimethylformamide in vacuo the residue was chromatographed on neutral alumina (0.5% methanol/CH₂Cl₂ - 5% methanol/CH₂Cl₂) to yield as a pale oil the compound of formula (1) wherein A is -(CH₂) p N(R)(CH₂) q -, m, n, p and q is each an integer 2; R¹, R², R³ and R⁴ is each a group -CO₂CH₂CH₃; X¹, X³ and X⁴ is each a hydrogen atom; and X² is a group -(CH₂)₄NHCOPh (wherer Ph is phenyl). m/e (DCE, ethanol) 692 (M⁺+1). δ H (CDCl₃) 1.25 (12H, t+t+t, Me), 1.40-1.90 (6H, mult, CH 2 CH 2 CH 2 CH₂NHCO), 2.13-3.20 (16H, mult, ring CH₂), 3.30-3.55 (9H, mult, CH₂CO+CHCO+CH₂NHCO), 4.08-4.18 (8H, mult., OCH₂) 7.40 (3H, brd, arom H), 7.94-8.07 (3H, brd + mult., ortho CH + NHCO). Example 2 A solution of the compound of Example 1(b) (30mg) in hydrochloric acid (6M, 10ml) was heated to reflux for 36h. After cooling, and washing with diethyl ether (3 x 3ml), the removal of solvent under high vacuum yielded the compound of formula (1) wherein A is -(CH₂) p N(R)(CH₂) q -, m, n, p and q is each an integer 2, R¹, R², R³ and R⁴ is each a group -CO₂H, X¹, X³ and X⁴ is each a hydrogen atom and X² is a group -(CH₂)⁴NH₂- as its tetrahydrochloride (28mg). ∂ H (D₂O) 1.64 (6H, br. mult., CH₂C), 2.93-4.16 (25H, mult. br., CH₂N+ CHN), m/e (negative FAB, m-nitrobenzyl alcohol) 476 (M⁻), 475 (M⁻-1). Example 3 To a solution of the compound of Example 2 (9.5mg) in dry DMF (200µl) was added a solution of N-succinimidyl-3-maleimidopropiorate (14.0mg) in dry DMF (60µl), and N-methyl morpholine (40mg) yielding a slight precipitate. Addition of MilliQ water (0.003ml) dissolved the precipitate and the mixture was held at 30°C for 24h. After removal of solvent under reduced pressure, the residue was purified on reverse-phase HPLC (Spherisorb 50DS2) to yield as a colourless glass and compound of formula (1) corresponding to the compound of Example 2 except that X² is a group -(CH₂)₄NHCO(CH₂)₂- m/e (FAB, m-nitribenzylalcohol) 628 (M⁺+1), 552, 493, 409. ∂ H (D₂O) 6.85 (2H,s, CH=CH), 3.96-3.91 (7H, mult, CH₂N+CHN), 3.85 3.05 (20H mult. br. CH₂N+CH₂NHCO+CH₂NCO), 2.50 (2H, t, J=6.4, CH₂CONH), 1.65 (6H, mult, CH₂C). Rt =8.9 min (Spherisorb ODS2, 1.4ml min⁻¹, t=0, 95% 0.1% TFA in H₂O, 5% 0.1% TFA in CH₃CN t=20, 5% 0.1% TFA in H₂O, 95% 0.1% TFA in CH₃CN). To a solution of the tetraacid (10µmol) in ammonium acetate buffer (90µl, pH6.0) was added 5µl³ of an ⁹⁰ Y solution (2µCi) and the mixture was incubated for 0.5h at 37°C. Reaction was quenched by addition of a 20 fold excess of diethylenetriamindpentaacetic acid (DTPA) and the relative amounts of 90Y complex and 90Y-DTPA determined by HPLC radiometry (AX-300 anion exchange: eluant 0.2M NH₄OAC, 10% CH₃CN). A radiolabelling yield of 77% was calculated. In the presence of a 500 fold excess of DTPA at 25°C the relative concentration of the radiolabelled complex was measured at 24, 48 and 72h and seen not to diminish (±2%). Example 4 To a solution of 1,4,7-triazacyclononane (0.104g) in dry dimethylformamide (5ml) was added potassium carbonate (0.117g) followed by 2-bromo-6-benzamidoethyl hexanoate (0.276g) as a solution in dimethylformamide (3ml). After stirring for 50h at 50°C, the cooled solution was filtered and solvent removed under reduced pressure to give a colourless oil, m/e (DCI, ethanol) 652 (M⁺+1); HPLC [CM300; 2 major peaks (λmax 235nm) at 2.4 and 4.7 min with t=0: A=70%, B=10%, C=20%,; t=10min: A=0%, B=80%, C=20% (A=H₂0, B=1M ammonium acetate, C=CH₃CN)]. To a solution of the crude oil in dry ethanol (2ml) was added caesium carbonate (0.163g) and ethyl bromoacetate (0.083g). The mixture was stirred for 20 hours at 60°C when the solution was cooled, filtered and solvent removed to yield a pale yellow residue which was chromatographed on neutral alumina (1% methanol/CH₂Cl₂) to give as a colourless oil [Rf=0.5(2.5% methanol/CH₂Cl₂)] the compound of formula (1) wherein A is -(CH₂) p , m, n and p is each an integer 2, R¹, R² and R³ is each a group -CO₂CH₂CH₃, X¹ and X² is each a group -(CH₂)₄NHCOPh (where Ph is phenyl) and X³ is a hydrogen atom. m/e (DCI, ethanol) 738 (M⁺+1), 621, 420. δ H (CDCl₃ 298K) 1.24 (9H, t, CH₃, J=7.1Hz), 1.29-1.68 (12H, mult., CH 2 CH 2 CH 2 CH₂NH), 2.77-3.46 (20H, mult., CH₂N, CHN), 4.11 (6H, q + q, CH₂0), 6.66 (2H, brs, NHCO), 7.42 (6H, mult., arom H), 7.79 (4H, dd, ortho CH). Example 5 The compound of formula (1) wherein A is -(CH₂) p -, m, n and p is each an integer 2, R¹, R² and R³ is each a group -CO₂CH₂CH₃, X¹ is a group -(CH₂)₄NHCOPh (where Ph is phenyl) and X² and X³ is each a hydrogen atom was prepared using the methods of Example 4 except 1,4,7-triazacyclononane and 2-bromo-6-benzamidoethyl hexanoate were used in a 2:1 molar ratio. The product was purified by column chromatography on neutral alumina to yield a colourless oil, Rf=0.4 (2% methanol/CH₂Cl₂). m/e (DCI, ethanol) 563 (M⁺+1). δ H (CDCl₃, 298K) 1.25 (1H, t + t, CH₃), 1.28-1.69 (6H, mult., CH 2 CH 2 CH 2 CH₂-NHCO), 2.76-3.02 (12H, mult., CH₂N ring), 3.22-3.54 (7H, mult., CH ₂NHCO + C H CO + CH₂C0), 4.13 (6H, q, CH₂0), 6.67 (1H, brs, NHCO), 7.40 (3H, mult. arom CH), 7.79 (12H, brd, ortho CH). Example 6 N, N ¹ -bis(2-(N-(3-maleimidopropanoyl)-4-aminobutyl) carboxymethyl)-N″-carboxymethyl-1, 4, 7-triazacyclononane , (10mg) was prepared from the compound of Example 4 (38mg) using a similar procedure to that described for the preparation of the compound of Example 3 from the compound of Example 1. m/e (FAB, p-NBA matrix) 748(M⁺+1) 596; δ H (D₂O) 6.85 (4H, S, CH=CH) 3.96 (4H, m, CH₂CO₂) 3.79 (8H, m, C H ₂NCO and C H ₂NHCO) 3.10-3.25 (12H, m, CH₂N) 2.49 (4H, t, J6.3H₂, CH₂CONH) 1.82 (3H, m, CH₂-C) 1.46 (3H, m CH₂-C). Example 7 N-(2-(N-(3-maleimidopropanoyl)-4-aminobutyl)carboxymethyl)-N′, N″-bis(carboxymethyl0-1, 4, 7-triazacyclononane , was prepared from the compound of Example 5 using a similar procedure to that described for the preparation of the compound of Example 3 from the compound of Example 1. m/e (FAB, p-NBA matrix) 526(M⁺+1) 453, 391, 307; δ H (D₂O) 6.85 (2H, S, CH=CH) 3.95 (5H, m, CH₂CO₂) 3.79 (4H, m, C H ₂NCO and C H ₂NHCO) 3.00-3.50 (12H, m, CH₂N) 2.49 (2H, t, J6.2H₂, CH₂CONH) 1.69 (3H, m, CH₂-C) 1.46 (3H, m CH₂-C).

Claims: A compound of formula (1): wherein A is a group -(CH₂) p - or -(CH₂) p N(R)(CH₂) q - where R is a group -CH(R⁴)X⁴; m, n, p, and q is each an integer 2; R¹, R², R³ and R⁴, which may be the same or different, is each a group -CO₂H, -P(R⁵)O₂H (where R⁵ is a hydrogen atom or a C₁₋₆ alkyl or alkoxy group), -PO₃H₂ or -CONR⁶R⁷ (where R⁶ and R⁷, which may be the same or different is each a hydrogen atom or a C₁₋₆ alkyl group); X¹, X², X³ and X⁴, which may be the same or different, is each a hydrogen atom or a C₁₋₆ alkyl group, or a linker group of the formula -L-Z wherein L is an optionally substituted straight or branched C₁₋₂₀ alkylene, C₂₋₂₀ alkenylene, or C₂₋₂₀ alkynylene chain, optionally interrupted by one or more -O- or -S- atoms or C₅₋₈ cycloalkylene, C₆₋₁₂ aromatic, C₅₋₁₀ heteroaromatic, N(R⁸)-, -CON(R⁸)- or -N-(R⁸)CO- groups (where R⁸ is a hydrogen atom or a C₁₋₆ alkyl group), and Z is a halogen atom or a group selected from OH, -SH, -NH₂, hydrazine (-NHNH₂), -N(CH₃)NH₂, -NHCONHNH₂, -NHCSNHNH₂, or phenylhydrazine, -NCO, -NCS, -COR¹³, [where R¹³ is a halogen atom or a N₃, C₁₋₆ alkoxy, C₆₋₁₂ aryloxy, imidyloxy or imidazolyloxy group], imide, a vinyl group of formula -Het¹-C (Het²) = CH₂ (where Het¹ and Het², which may be the same or different, is each a nitrogen containing heterocyclic group, or Het¹ is a nitrogen containing heterocyclic group and Het² is a hydrogen atom), or a dione of formula (where R¹⁴ is a C₁₋₄ alkyl group); with the proviso that at least one of X¹, X², X³ or X⁴ is a linker group; and metal complexes and/or salts thereof. A compound according to Claim 1 wherein one or two of X¹, X², X³ and X⁴ is a linker group and the remainder are C₁₋₆ alkyl groups or hydrogen atoms. A metal complex of a compound of formula (1) as defined in any of the preceding claims in which A is -(CH₂) p N(R)(CH₂) q , wherein the metal is di- or tripositive and has a coordination number 6 or greater. A metal complex according to Claim 3 wherein the metal is indium, yttrium, gallium, terbium, gadolinium or scandium. A metal complex according to Claim 4 wherein the metal is yttrium. A metal complex of a compound according to Claim 1 or 2 in which A is -(CH₂) p -, wherein the metal is di- or tripositive and has a coordination number from 2 up to 6. A metal complex according to Claim 6 wherein the metal is indium, gallium, cobalt or copper. A metal complex according to Claim 7 wherein the metal is indium. A compound of formula (1b) according to claim 1 wherein one or two of X¹, X² or X³ is a linker group as defined in claim 1 and the remainder are hydrogen atoms; and metal complexes and/or salts thereof. A compound of formula (1d) according to claim 1 wherein one or two of X¹, X², X³ or X⁴ is a linker group as defined in claim 1 and the remainder are hydrogen atoms; and metal complexes and/or salts thereof.

Inventors: BEELEY NIGEL ROBERT ARNOLD, MILLICAN THOMAS ANDREW, PARKER DAVID, BEELEY, NIGEL ROBERT ARNOLD, MILLICAN, THOMAS ANDREW, PARKER, DAVID
Applicants: CELLTECH THERAPEUTICS LTD, CELLTECH THERAPEUTICS LIMITED
Application year: 1990
Main classification: C07D255
IPC codes: C07D257, A61P43, C07D403, A61K31, C07D255