RADIOPHARMACEUTICAL CONJUGATE

Abstract

This invention relates new radiopharmaceutical conjugates for use in improved methods of diagnosis and treatment of cancer. The radiopharmaceutical conjugate comprises, in sequence: a metabolite that targets tumour cells, bound to a chelating agent capable of containing a radionuclide, bound to a linker capable of binding with an EPR agent in vitro or in vivo; or a chelating agent capable of containing a radionuclide, bound to a metabolite that targets tumour cells, bound to a linker capable of binding with an EPR agent in vitro or in vivo. The radiopharmaceutical conjugates of the present invention provide active and passive targeted radio nuclide delivery systems that can help to improve the biodistribution and pharmacological toxicity of the radiopharmaceuticals used for the diagnosis and therapy of cancer.

Claims

1-36. (canceled)

37. A molecule comprising a metabolite that targets tumour cells, a chelating agent capable of containing a radionuclide, and a cleavable linker capable of binding with an EPR agent, or a cleavable linker bound to an EPR agent.

38. The molecule claimed in claim 37, wherein the molecule is a linear molecule comprising, in sequence: a metabolite that targets tumour cells, bound to a chelating agent capable of containing a radionuclide, bound to a cleavable linker capable of binding with an EPR agent, or a cleavable linker bound to an EPR agent .

39. The molecule claimed in claim 37, wherein the cleavable linker is bound to the EPR agent.

40. The molecule claimed in claim 37, wherein the cleavable linker comprises a carbon chain of 4 to 20 carbon atoms.

41. The molecule claimed in claim 40, wherein the cleavable linker comprises a carbon chain of 8 to 15 carbon atoms.

42. The molecule claimed in claim 37, wherein the cleavable linker is cleaved in vivo within a tumour environment.

43. The molecule claimed in claim 42, wherein the cleavable linker contains a hydrazone bond, a disulfide bond, or enzymatically cleavable peptide sequences.

44. The molecule claimed in claim 37, wherein the EPR agent is a molecule with a size greater than 40 kDa.

45. The molecule claimed in claim 44, wherein the EPR agent is selected from polymeric nanoparticles, polymeric micelles, dendrimers, liposomes, viral nanoparticles, carbon nanoparticles, and proteins that accumulate in a tumour in vivo due to the Enhanced Permeability and Retention (EPR) effect.

46. The molecule claimed in claim 45, wherein the EPR agent is a synthetic polymer that is biodegradable, a synthetic polymer that is biocompatible but not biodegradable, or a natural polymer.

47. The molecule claimed in claim 46, wherein the biodegradable synthetic polymer is polyglutamate (PG), polylactide (PLA) or poly(D,L-lactide-co-glycolide)(PLGA).

48. The molecule claimed in claim 46, wherein the polymer that is biocompatible but not biodegradable is Polyethylene glycol (PEG) or N-(2-hydroxypropyl) methylacrylamide (HMPA).

49. The molecule claimed in claim 46, wherein the natural polymer is albumin, chitosan or heparin.

50. The molecule claimed in claim 37, wherein the metabolite that targets tumour cells is a small molecule less than 1000 Da in size.

51. The molecule claimed in claim 50, wherein the metabolite that targets tumour cells is 100 to 700 Da in size.

52. The molecule claimed in claim 51, wherein the metabolite that targets tumour cells is selected from monoclonal antibodies, proteins, peptides and molecules that are in vivo recognized by up-regulated receptors, antigens or proteins on the cancer-cell surface.

53. The molecule claimed in claim 52, wherein the monoclonal antibody (mAb) is 2C5, Gemtuzumab, Rituximab, Cetuximab, Bevacizumab, Pertuzumab and PSMA Ab antibody.

54. The molecule claimed in claim 52, wherein the protein or peptide is Transferrin, RGD peptides, Octreotide, Bombesin, or VIP.

55. The molecule claimed in claim 52, wherein the small molecule is folate, mannose, glucose or galactose.

56. The molecule claimed in claim 37, wherein the chelating agent is a cyclic or acyclic bifunctional chelating agent (BFCA) which is able to complex a radioisotope.

57. The molecule claimed in claim 56, wherein the chelating agent is a cyclic chelator selected from:1,4,7-Triazacyclononane (TACN); 1,4,7-triazacyclononane-triacetic acid (NOTA); 1,4,7-triazacyclononane-N-succinic acid-N′,N″-diacetic acid (NOTASA); 1,4,7-triazacyclononane-N-glutamic acid-N′,N″-diacetic acid (NODAGA);1,4,7-triazacyclononane-N,N′,N″-tris (methylenephosphonic) (NOTP); 1,4,7,10-tetraazacyclododecane ([12]aneN4) (cyclen); 1,4,7,10-tetraazacyclotridecane ([13]aneN4); 1,4,7,11-tetraazacyclotetradecane (iso-cyclam); 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA); 2-(1,4,7,10-tetraazacyclododecan-1-yl)acetate (DO1A); 2,2′-(1,4,7,10-tetraazacyclododecane-1,7-diyl) diacetic acid (DO2A); 2,2′,2″-(1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid (DO3A); 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methanephosphonic acid) (DOTP); 1,4,7,10-tetraazacyclododecane-1,7-di(methanephosphonic acid) (DO2P); 1,4,7,10-tetraazacyclododecane-1,4,7-tri(methanephosphonic acid) (DO3P); 1,4,7,10-tetraazacyclo-decane-1-glutamic acid-4,7,10-triacetic acid (DOTAGA); 1,4,7,10-tetraazacyclodecane-1-succinic acid-4,7,10-triacetic acid (DOTASA); 1,4,8,11-tetraazacyclotetradecane ([14]aneN4) (cyclam); 1,4,8,12-tetraazacyclopentadecane ([15]aneN4); 1,5,9,13-tetraazacyclohexadecane ([16]aneN4); 1,4-ethano-1,4,8,11-tetraazacyclo-tetradecane (et-cyclam); 1,4,8,11-15-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA); 2-(1,4,8,11-tetraazacyclotetradecane-1-yl) acetic acid (TE1A); 2,2′-(1,4,8,11-tetraazacyclotetradecane-1,8-diyl) diacetic acid (TE2A); 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]-hexadecane (CB-TE2A); 3,6,10,13,16,19-hexaazabicyclo[6.6.6]icosane (Sar); phthalocyanines and their derivatives; porphyrins and their derivatives.

58. The molecule claimed in claim 56, wherein the chelating agent is an acyclic chelator selected from: ethylene-diamine-tetraacetic-acid (EDTA); and diethylene-triamine-penta-acetic acid (DTPA). S-acetylmercaptosuccinic anhydride (SAMSA); (2-mercaptoethyl)(2-((2-mercaptoethyl)amino)ethyl)-carbamic acid (N2S2-DADT); 1,1′-(ethane-1,2-diylbis(azanediyl))bis(2-methylpropane-2-thiol) (N2S2 BAT-TM), (2-(2-mercaptoacetamido)ethyl)-cysteine (N2S2-MAMA); 2,3-bis(2-mercaptoacetamido)-propanoic acid (N2S2 DADS); ethylenedicysteine (EC);2,2′,2″-nitrilotriethanethiol (NS3); 2-ethylthio-N,N-bis(pyridin-2-yl)methyl-ethanamine (N3 S); ((2-mercaptoacetyl)glycylglycyl)carbamic acid (MAG3) and 4-(2-(2-(2-mercaptoacetamido)acetamido)-acetamido)butanoic acid (MAG2-GABA); (1,2-bis{[[6-(carboxy)pyridine-2-yl]methyl]-amino}-ethane) (H2dedpa); Nitrilotris(methylenephosphinic acid) (NTMP); ethylenediaminetetramethylene-phosphonic acid (EDTMP), diethylenetriaminepenta-methylene phosphonic acid (DTPMP); Hydrazinonicotinic acid (HYNIC); N′-{5-[Acetyl(hydroxy)amino]-pentyl}-N-[5-({4-[(5-aminopentyl)-(hydroxy)amino]-4-oxobutanoyl} amino)pentyl]-N- hydroxysuccin-amide (Deferoxamine).

59. The molecule claimed in claim 37, wherein the radionuclide is a radionuclide that may be used for imaging selected from: .sup.99mTc, .sup.188Re, .sup.186Re, .sup.153Sm, .sup.67Ga, .sup.68Ga, .sup.111In, .sup.59Fe, .sup.63.sub.Zn, .sup.52.sub.Fe, .sup.45Ti, .sup.60Cu, 61Cu, .sup.67Cu, .sup.64Cu, .sup.62Cu, .sup.198Au, .sup.199Au, .sup.195mPt, .sup.191mPt, .sup.193mPt, .sup.117mSn, .sup.89Zr, .sup.177Lu, .sup.18.sub.F, .sup.123I.

60. The molecule claimed in claim 37, wherein the radionuclide is a radionuclide that may be used for therapeutic purposes selected from: .sup.188Re, .sup.186Re, .sup.153Sm, .sup.166Ho, .sup.90Y, .sup.89Sr, .sup.111In, .sup.153Gd, .sup.225Ac, .sup.212Bi, .sup.213Bi, .sup.211At, .sup.60Cu, .sup.61Cu, .sup.67Cu, .sup.64Cu, .sup.62Cu, .sup.198Au, .sup.199Au, .sup.195mPt, .sup.193mPt, .sup.197Pt, .sup.117mSn, .sup.103Pd, .sup.103mRh, .sup.177Lu, .sup.223Ra, .sup.224Ra, .sup.227Th, .sup.32P, .sup.161Tb and .sup.33P, .sup.125I, .sup.203Pb, .sup.201Tl, .sup.119Sb, .sup.58mCo, .sup.161Ho.

61. The molecule claimed in claim 37, wherein the radionuclide is an Auger electron emitting radionuclide.

62. The molecule claimed in claim 61, wherein the Auger electron emitting radionuclide is selected from .sup.111In, .sup.203Pb, .sup.201Ti, .sup.103Pd, .sup.103mRh, .sup.119Sb, .sup.58mCo, .sup.161Ho, .sup.161Tb, .sup.61Cu, .sup.67Cu, .sup.195mPt, .sup.193Pt, .sup.117mSn.

63. The molecule claimed in claim 37, wherein the chelating agent contains the radionuclide.

64. The molecule as claimed in claim 37, wherein: the metabolite that targets tumour cells is a glucose-containing linker that is functionalised for connection to the chelating agent through alkylation or acylation; the chelating agent is a cyclam functionalised through N-linkages for radioisotope chelation; the linker is functionalised with maleimide; and the EPR agent is albumin.

65. The molecule as claimed in claim 64, further comprising .sup.103Pd.

66. A method for the synthesis of a molecule as defined in claim 37 includes the steps of: functionalization of a metabolite that targets tumour cells, wherein the metabolite is reacted with an alkyl halide chain to form a metabolite connected to a carbon chain with a terminal functional group that is then converted into a halide or an acid chloride; functionalization of a cleavable linker, wherein two fragments with terminal functional groups are connected through a bond that can be cleaved, the first fragment containing an alkyl halide at one end for attachment to the chelating agent and a suitable group to form the cleavable bond at the other terminus and the second fragment with a protected amine at the one end and a suitable group to react with the first fragment to form the cleavable bond at the other end; functionalization of a chelating agent, wherein the chelating agent is first mono-alkylated with the linker and then alkylated a second time with the metabolite, and wherein the remaining amines of the macrocycle are reacted with acetate groups that assist in metal complexation, and wherein the terminal amine is then converted into a functional group capable of binding to an EPR agent.

Description

EXAMPLES

Example 1

[0122] Radiolabelled Metabolite-Chelator-Linker Pro-Conjugate for Attachment to an EPR Agent and Formation of a Radiopharmaceutical Bioconjugate

[0123] A) Two proposed synthetic glucose-cyclam-maleimide pro-conjugates with various linker alternatives that are radiolabelled with .sup.103Pd. B) Michael addition of the free thiol in albumin to the maleimide of the pro-conjugates to form the radiopharmaceutical bioconjugate.

Example 2

Metabolite—Synthesis of Glucose Linker

[0124] 10-(tent-butyldiphenylsilyloxy)decan-1-ol (1)

##STR00004##

[0125] Imidazole (3.50 g, 51.7 mmol), followed by TBDPSCI (8.70 g, 31.5 mmol) was added slowly to a solution of 1,10-decanediol(5.00 g, 28.7 mmol) in dry THF (60 mL) under N.sub.2(.sub.g) and left stirring at room temperature for 24 hrs. The reaction was quenched by evaporation of THF in vacuo followed by addition of water (60 mL) and CH.sub.2Cl.sub.2 (60 mL). The organic phase was separated and extracted with water (2×50 mL) and washed with brine (50 mL). The organic phase was dried, filtered and concentrated. The crude product was columned [Hexane: EtOAc (9:1)] and pure alcohol obtained as an oil (6.59 g, 56%). R.sub.f=0.42 (Hexane: EtOAc 8:2).

2,3,4,6-Tetra-O-benzoyl-α-D-glucopyranosyl Benzoate (2)

[0126] ##STR00005##

[0127] Benzoyl chloride (4.68 g, 33.3 mmol) was added dropwise to a solution of α-D-glucose (1.0 g, 5.6 mmol) in pyridine (30 mL) at 0° C. After 10 min at 0° C., the solution was stirred for 2 hrs at room temperature. The reaction was quenched by addition of cold water (50 mL) and the product extracted with EtOAc (3×50 ml). The combined organic layers were washed with 1N HCl (3×50 mL) followed by brine (50 mL) and then dried, filtered and concentrated. The crude product was purified by recrystallisation with hot Hexane: EtOAc 2:1 to give the title compound as a white solid (3.48 g, 89%). R.sub.f=0.28 (Hex: EtOAc 8:2). δ.sub.H (CDCl.sub.3, 400 MHz): 8.16 (2H, d, J=8.0 Hz, H-Ar), 8.02 (2H, d, J=8.0 Hz, H-Ar), 7.94 (2H, d, J=8.0 Hz, H-Ar), 7.88 (4H, d, J=8.0 Hz, H-Ar), 7.66 (1H, t, J=8.0 Hz, H-Ar), 7.53-7.28 (14H, m, H-Ar), 6.85 (1H, d, J=4.0 Hz, H-1), 6.32 (1H, t, J=8.0 Hz, H-3), 5.85 (1H, t, J=8.0 Hz, H-4), 5.68 (1H, dd, J=4.0, 8.0 Hz, H-2), 4.62 (2H, m, H-6a/H-5), 4.59 (1H, dd, J=4.0 Hz, J=12.0 Hz, H-6b)

δ.sub.C (CDCl.sub.3, 100 MHz): 166.1, 165.9, 165.3, 165.1, 164.4 (C═O), [133.9, 133.5, 133.4, 133.3, 133.1 130.0, 129.9 (x2), 129.8 (x2), 129.6, 129.0, 128.9, 128.8, 128.6, 128.4 (x3), 128.37 ArC)], 90.0 (C-1), 76.6 (C-3), 70.5 (C-2), 70.5 (C-5), 68.9 (C-4), 62.5 (C-6)

1-Iodo-2,3,4,6-Tetra-O-benzoyl-α-D-glucopyranoside (3)

[0128] ##STR00006##

[0129] Hexamethyldisilane (0.531 g, 3.63 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added to a solution of α-D-glucose-pentabenzoate(2) (4.10 g, 5.85 mmol) in CH.sub.2Cl.sub.2 (60 mL). To this solution was added ZnI.sub.2 (0.467 g, 1.46 mmol), followed by I.sub.2 (0.921 g, 3.63 mmol) and stirred for 16 hrs. The reaction was quenched by addition of CH.sub.2Cl.sub.2 (40 mL) and an aqueous solution (120 mL) of NaHCO.sub.3 (1.68 g) and Na.sub.2S.sub.2O.sub.3 (1.12 g) and then stirring for 10 min until the pinkish colour and milky solution had cleared. The organic phase was separated and washed with brine (50 mL) and the combined aqueous phases extracted with CH.sub.2Cl.sub.2 (2×50 mL). The combined organic extracts were dried over MgSO.sub.4, filtered and concentrated in vacuo to yield a crude oil of the title compound which was used directly in the next reaction.

10-bromodecyl-tetra-O-benzoyl-β-D-glucopyranoside (4)

##STR00007##

[0130] Previously prepared iodide 3 (1.81 g, 2.57 mmol) was dissolved in CH.sub.2Cl.sub.2 (40 mL) under N.sub.2(g) and 4 Å Molecular Sieves (4.00 g) along with ZnCl.sub.2 (0.525 g, 3.85 mmol), and 10-bromodecanol (0.914 g, 3.85 mmol) in CH.sub.2Cl.sub.2 (5 mL) were added to the solution. The reaction was stirred for 17 hrs after which the colour of the solution had changed to a light pink. EtOAc (60 mL) was added and the reaction was quenched by addition of an aqueous solution (80 mL) of NaHCO.sub.3(.sub.s) (1.80 g) and Na.sub.2S.sub.2O.sub.5(.sub.s) (2.40 g). The colour changed from a yellow-orange to a milky white after stirring for 10 min. The solution was filtered through a Celite pad and the phases separated. The organic phase was washed with brine and the combined aqueous phases extracted with EtOAc (2×30 mL). The combined organic layers were dried, filtered and concentrated to yield a crude oil (2.99 g), which was purified by column chromatography (Hexane: EtOAc 8:2). The title product was obtained as a clear oil (1.23 g, 59% over 2 steps). R.sub.f=0.48 (Hex: EtOAc 8:2).

[0131] δ.sub.H (CDCl.sub.3, 400 MHz): 8.0-7.80 (8H, m, ArH), 7.53-7.24 (12H, m, ArH), 5.89 (1H, t, J=9.6 Hz, H-3′), 5.65 (1H, t, J=9.6 Hz, H-4′), 5.49 (1H, dd, J=7.8, 9.7 Hz, H-2′), 4.81 (1H, d, J=7.8 Hz, H-1′), 4.61 (1H, dd, J=3.3, 12.0 Hz, H-6a′), 4.49 (1H, dd, J=5.2, 12.0 Hz, H-6b′), 4.13 (1H, m, H-5′), 3.89 (1H, dt, J=6.3, 9.6 Hz, H-1a), 3.52 (1H, dt, J=6.7, 9.6 Hz, H-1b), 3.37 (2H, t, J=6.6 Hz, H-10), 1.80 (2H, qn, J=9.6 Hz, H-9), 1.55-1.45 (2H, m, H-2), 1.41-1.30 (2H, m, Alk-H), 1.19-1.05 (10H, m, Alk-H). δ.sub.C (CDCl.sub.3, 100 MHz): [166.2, 165.8, 165.2, 165.1 (C═O)], [133.4, 133.2, 133.1, 133.0, 129.8 (x2), 129.7 (x2), 129.6, 129.5, 128.9, 128.8, 128.4, 128.3, 128.3, 128.2 (ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.2 (C-5′), 72.0 (C-2′), 70.3 (C-1), 69.9 (C-4′), 63.3 (C-6′), 33.9 (C-10), 32.8, 29.4, 29.3, 29.2, 29.1, 28.7, 28.1, 25.7 (C-Alk).

10-(tert-butyldiphenylsilyloxy)decyl-tetra-O-benzoyl-β-D-glucopyranoside (5)

##STR00008##

[0132] Molecular sieves (7.00 g), ZnCl.sub.2 (1.43 g, 10.5 mmol) and alcohol 1 (2.17 g, 5.25 mmol) in CH.sub.2Cl.sub.2 (15 mL) was added to freshly prepared iodide 3 (3.71 g, 5.25 mmol) in CH.sub.2Cl.sub.2 (60 mL) and the reaction stirred for 8 hrs. CH.sub.2Cl.sub.2 (40 mL) was added to the solution and the molecular sieves were filtered off through a celite pad followed by the addition of an aqueous solution (100 mL) of NaHCO.sub.3 (0.960 g) and Na.sub.2S.sub.2O.sub.3 (1.44 g) and stirred for 10 min. The organic layer was separated and the aq. phase extracted with CH.sub.2Cl.sub.2 (3×50 mL). The combined organic layers were washed with brine and aq. phase extracted once more with CH.sub.2Cl.sub.2 (50 mL). The organic extracts were dried over MgSO.sub.4, filtered and concentrated in vacuo to yield a crude oil product which was dry-loaded onto a prepacked column and purified using column chromatography (Hex:EtOAc 9:1, 8:2, 7:3) to yield the title compound as an oil product (3.55 g, 68%). R.sub.f=0.51 (Hex: EtOAc 8:2) δ.sub.H (CDCl.sub.3, 400 MHz): 8.05-7.83 (8H, m, ArH), 7.68 (4H, m, ArH), 7.55-7.26 (18H, m, ArH), 5.91 (1H, t, J=9.6 Hz, H-3′), 5.68 (1H, t, J=9.6 Hz, H-4′), 5.53 (1H, dd, J=7.8, 9.6 Hz, H-2′), 4.85 (1H, d, J=7.8 Hz, H-1′), 4.65 (1H, dd, J=3.3, 12.0 Hz, H-6a′), 4.52 (1H, dd, J=5.2, 12.0 Hz, H-6b′), 4.17 (1H, m, H-5′), 3.92 (1H, dt, J=6.3, 9.6 Hz, H-1a), 3.66 (2H, t, J=6.6 Hz, H-10), 3.55 (1H, dt, J=6.7, 9.6 Hz, H-1b), 1.59-1.50 (4H, m, H-2/9), 1.29 (2H, m, H-8), 1.22-1.00 (10H, m, AlkCH.sub.2), 1.04 (9H, s, H-2″).

δ.sub.C (CDCl.sub.3, 100 MHz): [166.1, 165.8, 165.2, 165.0 (C═O)], [135.6 (x4), 134.2 (x2), 133.4, 133.2, 133.1, 133.0, 129.8 (x2), 129.7 (x6), 129.5 (x2), 128.9 (x2), 128.4 (x2), 128.3 (x4), 128.3 (x2), 128.3 (x2), 127.5 (x4) (ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.2 (C-2′), 72.0 (C-5′), 70.3 (C-1), 70.0 (C-4′), 64.0 (C-10), 63.3 (C-6′), 32.6, 29.5, 29.4, 29.4, 29.3, 29.2, 26.9 (C-2″), 25.8, 25.7, 19.2 (C-1″).
10-hydroxydecyl-tetra-O-benzoyl-β-D-glucopyranoside (6)

##STR00009##

[0133] Acetic acid (0.245 g, 4.09 mmol) and N-tetra butyl ammonium fluoride (6.80 mL, 1.0 M in THF, 6.80 mmol) was added to a solution of TBDPSO-C10-glucopyranoside 5 (3.38 g, 3.40 mmol) in THF (100 mL) and stirred for 36 hrs. The solvent was evaporated to yield a residue to which H.sub.2O (50 mL) was added and then extracted with EtOAc (3×50 mL). The combined organic extracts were washed with brine (50 mL), dried over MgSO.sub.4, filtered and concentrated. The crude material was purified with column chromatography (Hex:EtOAc 2:1) to yield the title alcohol as a clear oil (2.30 g, 90%) R.sub.f=0.16 (Hex: EtOAc 2:1), δ.sub.H (CDCl.sub.3, 300 MHz): 8.02-7.81 (8H, m, ArH), 7.56-7.25 (12H, m, ArH), 5.90 (1H, t, J=9.6 Hz, H-3′), 5.67 (1H, t, J=9.6 Hz, H-4′), 5.51 (1H, dd, J=7.8, 9.6 Hz, H-2′), 4.83 (1H, d, J=7.8 Hz, H-1′), 4.63 (1H, dd, J=3.3, 12.0 Hz, H-6a′), 4.51 (1H, dd, J=5.2, 12.0 Hz, H-6b′), 4.18-4.12 (1H, m, H-5′), 3.90 (1H, dt, J=6.3, 9.6 Hz, H-1a), 3.62 (2H, t, J=6.6 Hz, H-10), 3.53 (1H, dt, J=6.7, 9.6 Hz, H-1b), 1.63-1.46 (4H, m, AlkCH.sub.2), 1.35-1.05 (12H, m, AlkCH.sub.2).

δ.sub.C (CDCl.sub.3, 100 MHz): [166.1, 165.8, 165.2, 165.1 (C═O)], [133.4, 133.2, 133.1, 133.0, 129.8, 129.7, 129.6, 129.4, 128.9, 128.4, 128.3, 128.2 (ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.2 (C-2′), 72.0 (C-5′), 70.3 (C-1), 69.9 (C-4′), 63.3 (C-6′), 63.1 (C-10), 32.8, 29.4, 29.4, 29.3, 29.3, 29.1, 25.7, 25.6.
10-(tetra-O-benzoyl-β-D-glucopyranos-1-yl)-decanal (7)

##STR00010##

[0134] Dess-Martin periodinane (0.59 g, 1.4 mmol) was added to a solution of alcohol 6 (0.870 g, 1.16 mmol) in anh. CH.sub.2Cl.sub.2 (60 mL) and stirred for 1.5 hrs. The reaction was quenched by the addition of sat. aq. NaHCO.sub.3 solution (60 mL) and stirred for 15 min. The aqueous phase was extracted with CH.sub.2Cl.sub.2 (3×50 mL). The combined organic phase was washed with brine (50 mL), dried over MgSO.sub.4, filtered and concentrated. The crude material was purified with column chromatography (Hex:EtOAc 6:4) to yield the title compound as a clear oil (0.693 g, 79%), R.sub.f=0.75 (Hex: EtOAc 1:1). δ.sub.H (CDCl.sub.3, 300 MHz): 9.75 (1H, t, J=1.5 Hz, H-1), 8.02-7.82 (8H, m, ArH), 7.56-7.23 (12H, m, ArH), 5.90 (1H, t, J=7.2 Hz, H-3′), 5.67 (1H, t, J=7.2 Hz, H-4′), 5.51 (1H, dd, J=7.5, 6.0 Hz, H-2′), 4.83 (1H, d, J=6.0 Hz, H-1′), 4.63 (1H, dd, J=2.4, 9.0 Hz, H-6a′), 4.51 (1H, dd, J=4.2, 9.0 Hz, H-6b′), 4.14 (1H, m, H-5′), 3.91 (1H, dt, J=4.8, 7.2 Hz, H-10a), 3.54 (1H, dt, J=4.8, 7.2 Hz, H-10b), 2.38 (2H, dt, J=1.2, 5.4 Hz, H-2), 1.60-1.47 (4H, m, AlkCH.sub.2), 1.27-1.05 (10H, m, AlkCH.sub.2).

δ.sub.C (CDCl.sub.3, 100 MHz): 202.7 (C-1), [166.1, 165.8, 165.2, 165.0 (C═O)], [133.3, 133.1, 133.1, 133.0, 129.8 (x2), 129.7 (x2), 129.4, 128.9, 128.4 (x2), 128.3 (x2), 128.2 (x2)(ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.2 (C-2′), 72.0 (C-5′), 70.2 (C-10), 69.9 (C-4′), 63.3 (C-6′), 43.8 (C-2), 29.3, 29.1, 29.1, 29.1, 29.0, 25.7, 22.0 (C-Alk).
10-(tetra-O-benzoyl-β-D-glucopyranos-1-yl)decanoic acid (8)

##STR00011##

[0135] An aqueous solution (4.0 mL) of NaClO.sub.2 (0.157 g, 1.7 mmol) and NaH.sub.2PO.sub.4 (0.208 g, 1.30 mmol) was added to a solution of aldehyde 7 (1.00 g, 1.30 mmol) and 2-methyl-2-butene (0.626 g, 8.90 mmol) in t-butanol (40 mL) and the solution stirred for 2.5 hrs until all the yellow colour had disappeared. The solvent was evaporated to yield a crude oil which was redissolved in CH.sub.2Cl.sub.2 (50 mL) and H.sub.2O (100 mL). The solution was acidified with 1M HCl (10 mL) and the aqueous phase extracted with CH.sub.2Cl.sub.2 (3×50 mL). The combined organic extracts were dried over MgSO.sub.4, filtered and concentrated. The crude material was purified with column chromatography (Hex:EtOAc 1:1) to yield the title compound as a clear oil (0.90 g, 89%) R.sub.f=0.39 (Hex: EtOAc 1:1) (CDCl.sub.3, 300 MHz): 8.03-7.81 (8H, m, ArH), 7.53-7.26 (12H, m, ArH), 5.91 (1H, t, J=9.6 Hz, H-3′), 5.67 (1H, t, J=9.6 Hz, H-4′), 5.52 (1H, dd, J=7.8, 9.6 Hz, H-2′), 4.83 (1H, d, J=7.8 Hz, H-1′), 4.64 (1H, dd, J=3.4, 12.1 Hz, H-6a′), 4.51 (1H, dd, J=5.2, 12.1 Hz, H-6b′), 4.19-4.13 (1H, m, H-5′), 3.91 (1H, dt, J=6.2, 9.7 Hz, H-10a), 3.54 (1H, dt, J=6.2, 9.7 Hz, H-10b), 2.32 (2H, t, J=7.4 Hz, H-2), 1.62-1.46 (4H, m, AlkCH.sub.2), 1.27-1.02 (10H, m, AlkCH.sub.2).

δ.sub.C (CDCl.sub.3, 100 MHz): 179.1 (C═O), [166.1, 165.8, 165.2, 165.0 (C═O)], [133.4, 133.2, 133.0, 129.8, 129.7, 129.6, 129.4, 128.9, 128.4, 128.3, 128.2 (ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.2 (C-2′), 72.0 (C-5′), 70.3 (C-10), 69.9 (C-4′), 63.3 (C-6′), 33.9 (C-2), 29.3, 29.1, 29.1, 29.0, 28.9, 25.7, 24.6 (C-Alk)
10-(tetra-O-benzoyl-β-D-glucopyranos-1yl)decanoyl chloride (9)

##STR00012##

[0136] A few drops of DMF were added to a solution of decanoic acid 8 (0.900 g, 1.17 mmol) in anh. CH.sub.2Cl.sub.2 (30 mL) under N.sub.2(g). The flask was placed at 0° C. and oxalyl chloride (0.11 mL, 1.29 mmol) was added dropwise to the solution. The reaction was left stirring for 1 hr after which the solvent was removed under vacuum. A small amount of toluene (5 mL) was added and again evaporated in vacuo and the residue was then dried under a vacuum pump for 10 min to remove any remaining oxalyl-Cl. The crude oil product was not characterised but used directly in the next acylation reaction.

Example 3

Linker—Synthesis of Non-Cleavable Linker

10-Hydroxydecyl-phthalimide (10)

[0137] ##STR00013##

[0138] Potassium phthalimide (3.50 g, 18.9 mmol) was added to a solution of 10-bromodecanol (4.48 g, 18.9 mmol) in DMF (50 mL). The mixture was heated at 100° C. for 20 hrs after which most of the DMF was distilled off. The remaining product was redissolved in CH.sub.2Cl.sub.2 and then washed with H.sub.2O (2×50 mL). The combined organic layer was dried over MgSO.sub.4, filtered and concentrated in vacuo to yield a crude solid which was purified using column chromatography (Hex: EtOAc 6:4). The title compound was obtained as a white solid (4.92 g, 97%). R.sub.f=0.45 (Hex: EtOAc 1:1). δ.sub.H (CDCl.sub.3, 300 MHz): 7.84-7.81 (2H, m, ArH), 7.70-7.68 (2H, m, ArH), 3.66 (2H, t, J=7.2 Hz, H-1), 3.62 (2H, t, J=6.8 Hz, H-10), 1.66 (2H, qn, J=8.0 Hz, H-2), 1.55 (2H, qn, J=8.0 Hz, H-9), 1.42 (1H, s, —OH), 1.32-1.25 (12H, m, Alk-CH.sub.2)

δ.sub.C (CDCl.sub.3, 100 MHz): 168.4 (C═O), 133.8 (Ar-3′), 132.2 (ArC-2′), 123.1 (ArC-4′), 63.0 (C-10), 38.0 (C-1), 32.8 (C-9), [29.4, 29.3, 29.3, 29.0, 28.5, 26.8, 25.6 (Alk-CH.sub.2)]
10-aminodecan-1-ol (11)

##STR00014##

[0139] Hydrazine hydrate (0.81 g, 0.78 mL, 25.3 mmol) was added to a solution of 10-hydroxydecyl-phthalimide (10) (4.50 g, 14.8 mmol) in EtOH (180 mL) and refluxed overnight. Starting material remained and so extra hydrazine (0.4 ml) was added and the solution again refluxed overnight. The solvent was evaporated and the crude solid product redissolved in CH.sub.2Cl.sub.2 (100 mL) and basified with 2M NaOH until all solid had dissolved. The organic layer was separated out and the aqueous phase extracted with CH.sub.2Cl.sub.2 (2×80 mL). The combined organic layer was dried over MgSO.sub.4, filtered and concentrated in vacuo. The crude white solid obtained was recrystallised (CH.sub.2Cl.sub.2: Hexane) to yield the title compound as a white solid (2.22 g, 88%). R.sub.f=0.07 (CH.sub.2Cl.sub.2: MeOH 9:1 δ.sub.H (CDCl.sub.3, 400 MHz): 3.60 (2H, t, J=6.6 Hz, H-1), 2.66 (2H, t, J=6.9 Hz, H-10), 1.54 (2H, qn, J=7.0 Hz, H-9), 1.41 (2H, qn, J=6.9 Hz, H-2), 1.35-1.24 (12H, m, H-3-8) Sc (CDCl.sub.3, 100 MHz): 63.0 (C-1), 42.3 (C-10), 33.9 (C-9), 32.9 (C-2), [29.5, 29.5, 29.4, 29.4, 26.9, 25.8 (Alk-CH.sub.2)]

O-Benzyl-N-(10-hydroxydecyl) carbamate (12)

##STR00015##

[0140] Na.sub.2CO.sub.3 (2.96 g, 27.9 mmol) and benzyl chloroformate (2.07 mL, 2.47 g, 14.5 mmol) was added to a solution of 10-amino-1-decanol (11) (1.93 g, 11.1 mmol) in CH.sub.2Cl.sub.2:H.sub.2O (1:1, 100 mL), which was stirred for 20 hrs. The organic layer was separated and aqueous layer extracted with CH.sub.2Cl.sub.2 (3×50 mL). The combined organic extracts were dried over MgSO.sub.4, filtered and concentrated in vacuo to yield a crude solid which was purified using column chromatography (Hex:EtOAc 6:4, 5:5) to give the title compound as a white solid (3.13 g, 91%). R.sub.f=0.35 (Hex: EtOAc 1:1),

δ.sub.H (CDCl.sub.3, 400 MHz): 7.36-7.26 (5H, m, ArH), 5.10 (2H, s, H-2′), 4.70 (1H, b.s, —NH), 3.63 (2H, t, J=6.6 Hz, H-10), 3.18 (2H, m, H-1), 1.58-1.47 (4H, m, H-2/9), 1.30-1.20 (12H, m, H-3-8)

[0141] δ.sub.C (CDCl.sub.3, 100 MHz): 156.4 (C═O), [136.7, 128.5, 128.0 (ArC)], 66.6 (C-2′), 63.0 (C-10), 41.1 (C-1), 32.8 (C-9), 29.9 (C-2), [29.4, 29.4, 29.3, 29.2, 26.7, 25.7 (AlkCH.sub.2)]

O-Cert-Butyl-N-(10-hydroxydecyl) carbamate (13)

##STR00016##

[0142] Di-tent-butyl dicarbonate (1.05 g, 4.8 mmol) dissolved in CH.sub.2Cl.sub.2(5 mL) was added at 0° C. to a solution of 10-amino-1-decanol (11) (0.70 g, 4.0 mmol) in CH.sub.2Cl.sub.2:MeOH (4:1, 20 mL) and stirred for 2.5 hrs at 0-5° C. All the solvent was removed under vacume and the white solid residue was purified using column chromatography (Hex:EtOAc5:5; 4:6;3:7) to give the title compound as a white solid (0.84 g, 76%). R.sub.f=0.60 (Hex: EtOAc 4:6), 3.sub.H (CDCl.sub.3, 400 MHz): 4.51 (1H, b.s, —NH), 3.62 (2H, t, J=6.6 Hz, H-10), 3.08 (2H, m, H-1), 1.55 (2H, m, H-2), 1.46 (2H, m, H-9), 1.43 (9H, s, H-3′), 1.35-1.25 (12H, m, H-3-8)

O-Benzyl-N-(10-bromodecyl) carbamate (14)

##STR00017##

[0143] Triphenylphosphine (3.45 g, 13.15 mmol) and carbon tetrabromide (4.38 g, 13.15 mmol) were added to a solution of carbamate 12 (2.70 g, 8.77 mmol) in dry CH.sub.2Cl.sub.2 (160 mL) under N.sub.2(.sub.g), which was stirred for 3 hrs. Silica was added to the solution and the crude material was then dry loaded onto a prepacked column and purified using automated flash column chromatography (Hexane:EtOAc 9:1, 8.2). The title compound was obtained as a white solid (2.99 g, 92%). R.sub.f=0.68 (Hex: EtOAc 1:1);

δ.sub.H (CDCl.sub.3, 300 MHz): 7.35-7.28 (5H, m, ArH), 5.09 (2H, s, H-2′), 4.75 (1H, b.s, —NH), 3.40 (2H, t, J=6.8 Hz, H-10), 3.18 (2H, m, H-1), 1.84 (2H, qn, J=6.8 Hz, H-9), 1.50-1.36 (4H, m, H-2, H-8), 1.33-1.20 (10H, m, Alk-CH.sub.2) o.sub.c (CDCl.sub.3, 100 MHz): 156.4 (C═O), [136.7, 128.5, 128.3, 128.0 (Ar—C)], 66.5 (C-2′), 41.1 (C-1), 33.9 (C-10), 32.8 (C-9), 29.9 (C-2), [29.3, 29.3, 29.1, 28.7, 28.1, 26.7 (AlkC)]
O-tert-Butyl-N-(10-bromodecyl)carbamate (15)

##STR00018##

[0144] Triphenylphosphine (0.623 g, 2.37 mmol) and carbon tetrabromide (0.787 g, 2.37 mmol) were added to a solution of carbamate 13 (0.500 g, 1.82 mmol) in dry CH.sub.2Cl.sub.2 (20 mL) under N.sub.2(g), which was stirred for 2.5 hrs. Silica was added to the solution and the crude material was then dry loaded onto a column and purified using column chromatography (Hexane:EtOAc 9:1, 8.2). The title compound was obtained as a white solid (0.56 g, 91%). R.sub.f=0.65 (Hex: EtOAc 8:2);

δ.sub.H (CDCl.sub.3, 400 MHz): 4.49 (1H, b.s, —NH), 3.40 (2H, t, J=6.6 Hz, H-10), 3.09 (2H, m, H-1), 1.85 (2H, m, H-9), 1.46-1.38 (13H, m, H-2/8/3′), 1.33-1.25 (10H, m, H-3-7)

Example 4

Linker—Synthesis of Components for a Cleavable Linker

[0145] S-(10-((tert-butoxycarbonyl)amino)decyl) ethanethioate (16)

##STR00019##

[0146] Thioacetic acid (0.078 g, 1.03 mmol) was added to a suspension of NaH (60% in mineral oil) (0.034 mg, 0.86 mmol) in anhydrous THF (5 mL) and stirred for 10 min. This solution was then added dropwise to a solution of carbamate 15 (0.290 mg, 0.86 mmol) in anh. THF (5.0 mL) under N.sub.2(.sub.g) at 0° C. and stirred overnight at RT°. All solvent was evaporated under vacuum and the residue redissolved in EtOAc (10 mL) followed by extraction with sat. NH.sub.4Cl (.sub.aq) (2×15 mL). The organic phase was then washed with NaHCO.sub.3 (15.0 mL) and brine (15.0 mL). The organic phase was dried, filtered and concentrated and the crude oil purified with column chromatography (Hex:EtOAc 9.5:0.5) to yield the title compound as a white solid (0.178 mg, 62%), R.sub.f=0.40 (Hex: EtOAc 9:1)

δ.sub.H (CDCl.sub.3, 400 MHz): 4.49 (1H, b.s, —NH), 3.09 (2H, qt, J=6.0 Hz, H-1), 2.85 (2H, t, J=7.2 Hz, H-10), 2.31 (3H, s, H-2″), 1.56 (2H, qn, J=7.6 Hz, H-9), 1.46-1.40 (11H, m, H-2/311.35-1.26 (12H, m, H-3-8)
O-tert-butyl N-(10-mercaptodecyl)carbamate (17)

##STR00020##

[0147] NaOMe (25% in MeOH)(0.05 mL) was added to a solution of thioacetate 16(0.160 g, 0.48 mmol) in MeOH (2.0 mL) and stirred for 30 min. No starting material remained and so all solvent was evaporated and the residue redissolved in EtOAc followed by washing with NH.sub.4Cl.sub.(aq). The organic phase was dried, filtered and concentrated and purified with column chromatography (Hex: EtOAc 9:1) to yield the title compound as a white solid (0.135 g, 98%) R.sub.f=0.37 (Hex: EtOAc 9:1).

δ.sub.H (CDl.sub.3, 400 MHz): 4.49 (1H, b.s, —NH), 3.09 (2H, qt, J=6.4 Hz, H-1), 2.51 (2H, qt, J=7.2 Hz, H-10), 1.59 (2H, qn, J=7.2 Hz, H-9), 1.46-1.40 (11H, m, H-2/3′),1.36 (2H, m, H-8), 1.31 (1H, t, J=7.6 Hz), 1.27 (10H, m, H-3-7)

3-Bromopropyl-thiotosylate (18)

[0148] ##STR00021##

[0149] 1,3-dibromopropane (3.56 g, 17.64 mmol) was added to a solution of potassium thiotosylate (1.00 g, 4.41 mmol) in MeCN (30.0 mL) under N.sub.2 (g) and refluxed for 2 hrs. The reaction was then removed from the heat, allowed to cool and the solvent evaporated under vacuum. The residue was redissolved in CH.sub.2Cl.sub.2 (25.0 mL) and washed with water (3×20 mL) and brine (1×20 mL). The organic phase was then dried, filtered and concentrated followed by purification of the crude material with column chromatography (Hex:EtOAc 9:1). The title compound was obtained as a clear oil (1.02 g, 75%). R.sub.f=0.35 (Hex: EtOAc 8:2).

Example 5

Pro-Conjugate—Synthesis by Attachment of Glucose Linker and Non-Cleavable Linker to Cyclam

[0150] 1,4,8,11-Tetraazatricyclo[9.3.1.1]hexadecane (19)

##STR00022##

[0151] Formaldehyde (37% in H.sub.2O) (0.75 mL, 10.0 mmol) was added to a solution of cyclam (1.00 g, 5.0 mmol) dissolved in water (20 mL) and cooled in an ice bath to 0-5° C. The solution was stirred for 5 min at that temperature after which it was allowed to warm to room temperature and stirred for 2.5 hrs. The reaction was cooled again to 0-5 ° C. and stirred for 5 min to maximise the precipitation of white solid which had formed which was then filtered off and washed with ice water (3×20 mL). The solid was dissolved in CH.sub.2Cl.sub.2 (25 mL) and MeOH (5 mL) and the solution dried over MgSO.sub.4.

[0152] The MgSO.sub.4 was filtered of and the solvent removed in vacuo to yield the title product as a white solid (1.05 g, 94%).

δ.sub.H (CDCl.sub.3, 400 MHz): 5.40 (2H, dt, J=3.0 Hz, 13.5 Hz, H-1′a), 3.14 (4H, m, H-2/3/9/10), 2.90 (2H, d, J=13.5 Hz, H-1′b), 2.85-2.80 (4H, m, H-5/7/12/14), 2.62 (4H, td, J=4.5 Hz, 15.5 Hz, H-5/7/12/14), 2.38 (4H, m, H-2/3/9/10), 2.28-2.18 (2H, m, H-6/13), 1.21-1.16 (2H, dqn, J=2 Hz, J=16.5 Hz, H-6/13),
δ.sub.C (CDCl.sub.3, 100 MHz): 68.9 (C-1′), 53.6 (C-2/3/9/10), 49.3 (C-5/7/12/14), 20.3 (C-6/13)
1-[10-(Benzyloxycarbonylamino)decyl]-1,4,8,11-tetraaza-4,8-methano-cyclopentadecane ammonium bromide (20)

##STR00023##

[0153] Bromide 15 (1.06 g, 2.88 mmol) was added under N.sub.2(.sub.9) to a solution of bridged cyclam 19 (0.773 g, 3.45 mmol) in anh. CH.sub.3CN (80 mL) and stirred for 72 hrs in a flask wrapped with foil. The solvent was evaporated and the crude oil redissolved in CH.sub.2Cl.sub.2 (50 mL) and washed with H.sub.2O (50 mL). The aqueous layer was extracted with CH.sub.2Cl.sub.2 (2×50 mL) and the organic layers combined, dried, filtered and concentrated. The residue obtained was purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9:1 with few drops of NH.sub.4OH) to yield an oil (1.02 g, 69%), R.sub.f=0.56 (CH.sub.2Cl.sub.2: MeOH 9:1)

δ.sub.H (CDCl.sub.3, 400 MHz): 12.70 (1H, bs, NH), 9.10 (1H, bs, NH), 7.31-7.24 (5H, m, ArH), 5.05 (2H, s, H-2′), 4.77 (1H, bs, NH), 4.05 (1H, d, J=12.0 Hz, H-15a), 3.68 (1H, m, CH.sub.2N), 3.34 (1H, d, J=12.0 Hz, H-15b), 3.20-3.10 (4H, m, CH.sub.2N, H-25), 3.05-2.83 (8H, m, CH.sub.2N), 2.73 (1H, dt, J=5.0, 12.0 Hz, CH.sub.2N), 2.43 (1H, m, H-16a), 2.35-2.25 (3H, m, H-16b, CH.sub.2N), 2.20 (1H, m, CH.sub.2N), 2.11 (2H, m, CH.sub.2N, H-6a), 1.87 (1H, m, H-6b), 1.65 (1H, m, H-13a), 1.55 (1H, m, H-13b), 1.50-1.30 (4H, m, H-17/24), 1.28-1.20 (12H, m, CH.sub.2Alk)
δ.sub.C (CDCl.sub.3, 100 MHz): 156.4 (C-1′), [136.6, 128.4, 128.0, 128.0 (ArC)], 72.8 (C-15), 66.5 (C-2′), 53.7 (C-16), [52.9, 49.3, 48.5, 48.1, 48.0, 48.0, 46.0, 45.1 (C-N)], 41.0 (C-25), 29.9 (C-17), 29.5, 29.5, 29.4, 29.1, 27.7, 26.6, 26.3, 25.1 (C-13), 22.4 (C-6)
1-[10-(2,3,4,6-O-Tetrabenzoyl-β,D-glucopyranos-1-yl)-1-oxodecyl]-11-[10-(benzyloxycarbonylamino)decyl]-1,4,8,11-tetraaza-4,8-methano-cyclopentadecane (21)

##STR00024##

[0154] DMAP (0.046 g, 0.37 mmol) and Et.sub.3N (0.190 g, 1.88 mmol) was added to a solution of cyclam derivative 20 (0.460 g, 0.93 mmol) dissolved in anh. THF (35 mL). Acid chloride 9 (0.775 g, 1.0 mmol) dissolved in anh. THF (7.8 mL) was added to the solution and stirred for 1.5 hrs. The solution was filtered through Celite to remove triethylammonium salts followed by the removal of THF in vacuo. The crude oil was redissolved in CH.sub.2Cl.sub.2 (20 mL) and extracted with H.sub.2O (2×20 mL). The organic layer was washed with 2M NaOH (20 mL) and separated and the aqueous layer extracted with CH.sub.2Cl.sub.2 (2×20 mL). All organic phases were combined, dried, filtered and concentrated. The crude oil was purified using column chromatography (CH.sub.2Cl.sub.2: MeOH 9:1) to yield the title compound as an oil (0.76 g, 65%) R.sub.f=0.45 (CH.sub.2Cl.sub.2: MeOH 9:1). HPLC analysis indicated that the one spot contained two inseparable compounds, which were the title compound as indicated and the same compound with no bisaminal bridge.

δ.sub.H (CDCl.sub.3, 300 MHz): 8.01-7.81 (8H, m, ArH), 7.52-7.25 (17H, m, ArH), 5.89 (1H, t, J=9.6 Hz, 3′), 5.65 (1H, t, J=9.6 Hz, H-4′), 5.50 (1H, dd, J=7.8, 9.6 Hz, H-2′), 5.08 (2H, s, H-2″), 4.83 (1H, d, J=7.8 Hz, H-1′), 4.80 (1H, bs,NH), 4.62 (1H, dd, J=3.2, 12 Hz, H-6′a), 4.50 (1H, dd, J=5.2, 12.0 Hz, H-6′b), 4.15 (1H, m, H-5′), 3.90 (1H, dt, J=6.4, 9.6 Hz, H-25a), 3.85-3.55 (3H, bm, —NCH.sub.2), 3.53 (1H, dt, J=6.8, 9.6 Hz, H-25b), 3.41 (2H, bs, —NCH.sub.2), 3.17 (2H, t, J=6.8 Hz, H-35), 2.78-2.45 (10H, bm, —NCH.sub.2), 2.39-2.30 (5H, m, H-26/-NCH.sub.2), 2.26 (2H, m, H-17), 1.70-1.38 (12H, m, H-18/24/27/34/6/13), 1.35-1.00 (22H, m, CH.sub.2-alk),
δ.sub.C (CDCl.sub.3, 100 MHz): 172.9 (C═O), [166.1, 165.8, 165.2, 165.0 (C═O)], 156.4 (C═O), 136.7 (ArC), [133.4, 133.2, 133.2, 133.1 (ArC)], [129.8, 129.8, 129.7, 129.5, 128.9, 128.9, 128.5, 128.5, 128.4, 128.4, 128.3, 128.3, 128.1 (ArC)], 101.3 (C-1′), 73.0 (C-3′), 72.1 (C-2′), 71.9 (C-5′), 70.7 (C-15), 70.3 (C-25), 69.9 (C-4′), 66.5 (C-2″), 63.3 (C-6′), 56.0 (C-26), [55.4, 55.0, 54.7, 54.4, 54.1, 53.9, 52.7, 52.4, 51.9, 51.1, 50.2, 46.5, 45.1, 43.7 (NCH.sub.2)], 41.1 (C-35), 33.4/33.1 (C-17), 29.9 (C-34), 29.5, 29.5, 29.4, 29.4, 29.3, 29.2, 27.8, 27.6, 27.5, 26.9, 26.8, 25.7, 25.5, 21.6
1-[10-(2,3,4,6-O-Tetrabenzoyl-β,D-glucopyranos-1-yl)-1 -oxodecyl]-4,8-bis(tert-butoxycarbonylmethyl)-11-[10-(benzyloxycarbonylamino)decyl]-1,4,8,11 - tetraazacyclotetradecane (22)

##STR00025##

[0155] Potassium carbonate (0.033 g, 0.24 mmol) was added to a solution of cyclam 21 (0.100 g, 0.08 mmol) in anh. CH.sub.3CN (15 mL) under N.sub.2(.sub.9). t-Butylbrornoacetate(0.047 g, 0.24 mmol) was dissolved in anh. CH.sub.3CN (1 mL) and added to the solution which was then stirred for 16 hrs. The solvent was removed on the rotary evaporator and the residue was redissolved in H.sub.2O (20 mL) and extracted with CH.sub.2Cl.sub.2 (4×20 mL). The organic extracts were dried, filtered and the solvent evaporated under vacuum. The oily residue was purified using column chromatography (CH.sub.2Cl.sub.2: MeOH 9.5:0.5) to yield the title compound as an oil (0.086 g, 73%).

δ.sub.H (CDCl.sub.3, 400 MHz): 8.02-7.81 (8H, m, ArH), 7.53-7.25 (17H, m, ArH), 5.89 (1H, t, J=9.6 Hz, H-3′), 5.66 (1H, t, J=9.6 Hz, H-4′), 5.50 (1H, dd, J=7.8, 9.6 Hz, H-2′), 5.09 (2H, s, H-2″), 4.83 (1H, d, J=7.8 Hz, H-1′), 4.78 (1H, bs,NH), 4.63 (1H, dd, J=3.2, 12.0 Hz, H-6′a), 4.50 (1H, dd, J=5.2, 12.0 Hz, H-6′b), 4.15 (1H, m, H-5′), 3.90 (1H, dt, J=6.4, 9.6 Hz, H-25a), 3.53 (1H, dt, J=6.8, 9.6 Hz, H-25b), 3.45 (4H, m, NCH.sub.2), 3.25 (2H, m, H-1′″), 3.22 (2H, s, H-1″′), 3.17 (2H, t, J=6.8 Hz, H-35), 2.82-2.62 (8H, m, H-26/NCH.sub.2), 2.46 (2H, m, NCH.sub.2), 2.37 (4H, m, NCH.sub.2), 2.25 (2H, t, J=6.8 Hz, H-17), 1.67 (1H, bm, CH.sub.2), 1.57 (5H, m, CH.sub.2), 1.50 (4H, m, CH.sub.2), 1.45 (9H, s, H-4″), 1.43 (9H, s, H-4′″), 1.32-1.02 (24H, m, CH.sub.2Alk)
δ.sub.C (CDCl.sub.3, 100 MHz): 172.9 (C-16), 170.8 (C-2′″), [166.1, 165.8, 165.2, 165.0 (C═O)], 156.4 (C-1″), 136.7 (ArC), [133.3, 133.1, 133.1, 133.0, 129.8, 129.7 (x4), 129.4, 128.9 (x2), 128.5, 128.4, 128.3 (x3), 128.2, 128.0(ArC)], 101.3 (C-1′), 80.6 (x2)(C-3″), 73.0 (C-3′), 72.2 (C-2′), 72.0 (C-5′), 70.3 (C-25), 70.0 (C-4′), 66.5 (C-2″), 63.3 (C-6′), 57.2 (C-1″′), 55.9, (C-1″′), [55.0, 53.9, 52.3, 52.0, 51.3, 51.2 (x2), 47.0, 44.9 (NCH.sub.2)], 41.1 (C-35), 33.1/33.0 (0-17), [30.0, 29.5, 29.4 (x3), 29.3, 29.2 (x2), 28.2 (x2), 28.0, 27.6, 26.7 (x2), 26.5, 25.8, 25.5 (x3)(CH.sub.2Alk, C-6/13, C-4′″)]
1-[10-(β,D-glucopyranos-1-yl)-1-oxodecyl]-11-[10-(benzyloxycarbonylamino)decyl]-1,4,8,11-tetraaza-4,8-methano-cyclopentadecane (23)

##STR00026##

[0156] Sodium metal (0.119 g, 5.17 mmol) was reacted with anh. MeOH (5 mL) and then added to a solution of glucose-cyclam 21 (0.640 g, 0.512 mmol) in anh. MeOH (30 mL) under N.sub.2 (g) and stirred for 1 hr. The MeOH was evaporated in vacuo and the product redissolved in CH.sub.2Cl.sub.2 (20 mL). Water (20 mL) was added for extraction upon which an emulsion formed. The emulsion was left to separate and the organic layer was removed followed by further extraction of the aqueous phase with DCM (3×40 mL) with a bit of MeOH (5 mL). The organic layers were combined, dried, filtered and concentrated. The crude oil was dry-loaded and purified using automated column chromatography (CH.sub.2Cl.sub.2: MeOH: NH.sub.4OH, 8:1.8:0.2) to yield the title product as an oil (0.374 g, 92%). R.sub.f=0.55 (CH.sub.2Cl.sub.2: MeOH: NH.sub.4OH, 8:1.8:0.2)

δ.sub.H (CD.sub.3OD, 400 MHz): 7.34-7.23 (5H, m, ArH), 5.06 (2H, s, H-2″), 4.25 (1H, d, J=7.8 Hz, H-1′), 3.91-3.84 (2H, m, H-6′a/25a), 3.80-3.55 (2H, bm, —NCH.sub.2 including 3.67 (1H, m, H-6′b)), 3.60-3.40 (2H, bs, —NCH.sub.2 including 3.53 (1H, dt, J=6.9, 9.6 Hz, H-25b)), 3.37-3.23 (5H, m, H-3′/4′/NCH.sub.2), 3.17 (1H, t, J=7.8 Hz, H-2′), 3.10 (2H, t, J=7.2 Hz, H-35), 2.80-2.50 (10H, bm, —NCH.sub.2), 2.50-2.40 (4H, m, H-26/NCH.sub.2), 2.37 (2H, t, J=7.2 Hz, H-17), 1.74 (2H, m), 1.61 (4H, m), 1.48 (4H, m), 1.40-1.20 (24H, m, CH.sub.2-alk)
δ.sub.C (CD.sub.3OD, 100 MHz): 175.6 (C═O), 158.8 (C═O), 138.5 (ArC), [129.4, 128.9, 128.7 (ArC)], 104.4 (C-1′), 78.1 (C-3′), 77.9 (C-2′), 75.1 (C-5′), 71.7 (C-25), 71.1 (C-15), 70.9 (C-4′), 67.2 (C-2′), 62.8 (C-6′), 56.9 (C-26), [56.2, 55.4, 55.2, 55.2, 54.6, 54.5, 54.5, 53.8, 53.1, 52.5, 51.1, 51.1, 47.9, 46.0, 45.1, 42.3 (NCH.sub.2)], 41.8 (C-35), 34.2 /34.0 (C-17), [30.9, 30.8, 30.6, 30.6, 30.5, 30.5, 30.4, 30.4, 28.7, 28.6, 28.1, 28.0, 27.8, 27.0, 26.8, 26.7, 22.4 (C-alk including C-6/13)]
1-[10-(β,D-glucopyranos-1-yl)-1-oxodecyl]-11-[10-aminodecyl]-1,4,8,11-tetraazacyclotetradecane (24)

##STR00027##

[0157] Pd/C (0.037 g, 10% w/w) was added to a solution of glucose cyclam 23 (0.374 g, 0.45 mmol) in anh. MeOH (10 mL). The flask was flushed with H.sub.2(g) and stirred overnight under a H.sub.2 (g) environment using a hydrogen-filled balloon. The solution was filtered through a Celite pad which was washed with MeOH and the MeOH then removed under vacuum. The crude oil was then redissolved in MeOH (5 mL) and 2M NaOH (5 mL) added to obtain the product in its free base form. The water and MeOH were removed under vacuum and MeOH (10 mL) again added to the flask. A white solid precipitated out which was then filtered off though Celite. The solvent was evaporated and the crude product dry loaded and purified using column chromatography (CH.sub.2Cl.sub.2: MeOH: NH.sub.4OH, 7:2.5:0.5). Two major isomers were obtained which could not be fully separated (top isomer 0.040 g, mixed isomers 0.153 g, bottom isomer 0.084 g, total yield=89%). NMR analysis indicated that the top isomer still contained the bis-aminal bridge whereas the bottom isomer was analysed to be the title compound.

δ.sub.H (CD.sub.3OD, 400 MHz): 4.25 (1H, d, J=8.0 Hz, H-1′), 3.89 (1H, dt, J=6.8, 9.6 Hz, H-25a), 3.86 (1H, dd, J=2.0, 12.0 Hz, H-6′a), 3.68 (1H, dd, J=5.6, 12.0 Hz, H-6′b), 3.62-3.44 (5H, m, H-2/14/25b), 3.37 (1H, t, J=8.8 Hz, H-3′), 3.32-3.24 (2H, m, H-4′/5′), 3.17 (1H, t, J=8.0 Hz, H-2′), 2.96 (2H, m, —NCH.sub.2), 2.86-2.76 (8H, m, H-35/-NCH.sub.2(x3)), 2.66 (2H, m, -NCH.sub.2), 2.55-2.33 (6H, m, H-17/26/NCH.sub.2), 1.85 (2H, m, H-6), 1.80-1.55 (8H, m, H-13/18/24/34), 1.49 (2H, m, H-27), 1.42-1.28 (22H, m, CH.sub.2-alk)
δ.sub.C (CD.sub.3OD, 100 MHz): 175.6 (C-16), 104.4 (C-1′), 78.2 (C-3′), 77.9 (C-2′), 75.2 (C-5′), 71.8 (C-25), 70.9 (C-4′), 62.9 (C-6′), 56.0 (C-26), [53.5, 53.4, 52.8, 52.6, 51.3 (x2), 50.5 (x2), 49.7 (x2), 48.6 (x2), 48.0 (x2), 46.4 (x2) (NCH.sub.2 and rotamer)], 41.4 (C-35), 34.2/33.9 (C-17 and rotamer), [30.8, 30.6(x2), 30.4(x2), 30.4, 30.3, 28.7, 28.5, 28.2, 27.8, 27.6, 27.4, 27.0, 26.7 (CH.sub.2Alk including C-6/13 and rotamers)]
1-[10-(2,3,4,6-O-Tetrabenzoyl-(β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-1,4,8,11-tetraazacyclotetradecane (25)

##STR00028##

[0158] Bromide 4 (0.126 g, 0.16 mmol) was dissolved in CH.sub.3CN (5 mL) and added under N.sub.2(g) to a solution of 1,8-bis(t-butoxycarbonylmethyl)-1,4,8,11-tetraazacyclotetradecane (0.095 g, 0.22 mmol) in anh. CH.sub.3CN (5 mL) with K.sub.2CO.sub.3 (0.061 g, 0.44 mmol) and stirred for 72 hrs at 60° C. The solvent was evaporated and the crude oil redissolved in 1 M HCl (10 mL) and extracted with EtOAc (2×20 mL). The organic layers were combined, dried, filtered and concentrated and the residue was purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9.6:0.4; 9.4:0.6; 9.2:0.8; 9:1) to yield an oil (0.05 g, 28%), R.sub.f=0.45 (CH.sub.2Cl.sub.2: MeOH 9:1).

δ.sub.H (CDCl.sub.3, 400 MHz): 8.00-7.81 (8H, m, ArH), 7.53-7.25 (17H, m, ArH), 5.88 (1H, t, J=9.6 Hz, H-3′), 5.65 (1H, t, J=9.6 Hz, H-4′), 5.50 (1H, dd, J=7.8, 9.6 Hz, H-2′), 4.82 (1H, d, J=7.8 Hz, H-1′), 4.61 (1H, dd, J=3.2, 12.0 Hz, H-6′a), 4.50 (1H, dd, J=5.2, 12.0 Hz, H-6′b), 4.15 (1H, m, H-5′), 3.89 (1H, dt, J=6.4, 9.6 Hz, H-25a), 3.63 (2H, m, NCH.sub.2), 3.52 (1H, dt, J=6.8, 9.6 Hz, H-25b), 3.46 (2H, m, NCH.sub.2), 3.32 (2H, m, NCH.sub.2), 3.24-3.17 (2H, m,NCH.sub.2), 3.16-3.08 (4H, m, NCH.sub.2), 3.01 (2H, m, NCH.sub.2), 2.92 (2H, m, NCH.sub.2), 2.83 (2H, m, NCH.sub.2), 2.74 (2H, m, NCH.sub.2), 2.63 (2H, t, J=6.8 Hz, H-16), 2.05 (2H, m, H-6), 1.85 (2H, m, H-13), 1.53 (2H, m, CH.sub.2), 1.47 (9H, s, H-4″′), 1.43 (9H, s, H-4′″), 1.32-1.02 (12H, m, CH.sub.2Alk)
1-[10-(2,3,4,6-O-Tetrabenzoyl-β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-8-[10-(t-butoxycarbonylamino)decyl]-1,4,8,11- tetraazacyclotetradecane (26)

##STR00029##

[0159] Bromide 15 (0.029 g, 0.08 mmol) was added under N.sub.2(g) to a solution of cyclam 25 (0.050 g, 0.04 mmol) in anh. CH.sub.3CN (2 mL) with K.sub.2CO.sub.3 (0.018 g, 0.12 mmol) and stirred for 24 hrs at 60° C. The reaction was proceeding very slowly and so extra bromide 15 (0.019 g) and K.sub.2CO.sub.3 (0.012 g) was added and the reaction again stirred for 24 hrs at 60° C. The solvent was evaporated and the crude oil was redissolved in 1 M HCl (10 mL) and extracted with EtOAc (3×10 mL). The organic layers were combined, dried, filtered and concentrated and the residue was purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9.5:0.5 with few drops of AcOH) to yield an oil (0.03 g, 50%), R.sub.f=0.59 (CH.sub.2Cl.sub.2: MeOH 9.5:0.5).

1-[10-(β,D-glucopyranos-1-yl)]-4,11-diacetic acid-8-[10-aminodecyl]-1,4,8,11-tetraazacyclo tetradecane (27)

##STR00030##

[0160] TFA (1.0 mL) was added to a solution of cyclam 26 (0.030 g, 0.02 mmol) in anh. MeOH (4 mL) to make a final solution of 20% TFA. The reaction was stirred for 24 hrs at room temperature. The solvent and TFA was evaporated under vacuum to yield a residue which was used directly in the next reaction. The residue was dissolved in MeOH (2 mL) and was stirred for 5 min after addition of NaOMe (24% in MeOH)(0.5 mL). The reaction was quenched with the addition of Dowex H.sup.+(0.5 g) and stirred for 5 min afterwhich the Dowex was filtered off through a celite pad. The filtrate was concentrated to yield a solid residue to which MeOH (1 mL) was added to extract the product from the TFA salts. The MeOH was evaporated to yield a glassy solid (0.021 g) which still contained some TFA salt. LRMS: m/z calculated for C.sub.40H.sub.79N.sub.5O.sub.10=789.58; found (M+H.sup.+)=790.6

Example 6

Pro-Conjugate—Synthesis by Attachment of Glucose Linker and Cleavable Linker to Cyclam

[0161] 1-[10-(β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-1,4,8,11-tetraazacyclo-tetradecane (28)

##STR00031##

[0162] Sodium methoxide (0.20 mL, 25% solution) is added to a solution of cyclam25 (0.05 g, 0.04 mmol) in anh. MeOH (2.0 mL) and stirred for 30 min. The solvent is evaporated and the residue is redissolved in EtOAc (5 mL) and washed with 0.25 M HCl (2×5 mL). The organic phase is dried, filtered, concentrated and purified by column chromatography(CH.sub.2Cl.sub.2:MeOH 8.5:1.5) to yield the title compound as an oil.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-8-[3-(tosylthio)propyl]-1,4,8,11-tetraazacyclotetradecane (29)

##STR00032##

[0163] Bromide 18 (0.019 g, 0.06 mmol) is added under N.sub.2(.sub.g)to a solution of cyclam 28(0.020 g, 0.03 mmol) in anh. CH.sub.3CN (2 mL) with K.sub.2CO.sub.3 (0.012 g, 0.09 mmol) and stirred for 24 hrs at 60° C. The reaction is not quenched but proceeded directly to the next step.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-8-[3-(pent-4-yn-1-yldisulfanyl)propyl]-1,4,8,11-tetraazacyclotetradecane (30)

##STR00033##

[0164] 4-Pentyne-1-thiol (0.004 g, 0.04 mmol) is added to the previous reaction solution which is then refluxed for a further 2 hrs. The solvent is evaporated under vacuum and the crude residue was redissolved in EtOAc (5 mL) and washed with sat. aq. NH.sub.4Cl (5 mL). The organic phase is dried, filtered, concentrated and purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9:1) to yield the title compound as an oil.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-bis(tert-butoxycarbonylmethyl)-8-[3-((((tert-butoxycarbonyl)amino)decyl)disulfanyl)propyl]-1,4,8,11-tetraazacyclotetradecane (31)

##STR00034##

[0165] Thiol 17 (0.011 g, 0.04 mmol) is added to the solution from reaction 29 and is then refluxed for a further 2 hrs. The solvent is evaporated under vacuum and the crude residue is redissolved in EtOAc (5 mL) and washed with sat. aq. NH.sub.4Cl (5 mL). The organic phase is dried, filtered, concentrated and purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9:1) to yield the title compound as an oil.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-(diacetic acid)-8-[3-((aminodecyl)disulfanyl)propyl]-1,4,8,11-tetraazacyclotetradecane (32)

##STR00035##

[0166] A solution of 6 M HCl/EtOAc (0.50 mL) is added to Cyclam 31(0.020 g, 0.02 mmol) dissolved in EtOAc (4.5 mL) and stirred for 2 hrs. The product precipitated out as a white HCl salt which is then filtered off, washed once with EtOAc (5 mL) and dried.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-(diacetic acid)-8-[3-(pent-4-yn-1-yldisulfanyl)propyl]-1,4,8,11-tetraazacyclotetradecane (33)

##STR00036##

[0167] A solution of 6 M HCl/EtOAc (0.50 mL) is added to Cyclam 30 (0.020 g, 0.02 mmol) dissolved in EtOAc (4.5 mL) and stirred for 2 hrs. All solvent is evaporated off under vacuum and the solid product was recrystallised from EtOH.

Example 7

Radiolabelling—Synthesis and Radiolabelling of a Glucose-Cyclam Intermediate for Proof of Principle of .SUP.103.Pd Co-Ordination

[0168] 1,4,8-Tris-(tert-butoxycarbonylmethyl)-1,4,8,11-tetraazacyclotetradecane (34)

##STR00037##

[0169] t-Butylbromoacetate (0.205 g, 1.05 mmol) dissolved in CH.sub.3CN (25 mL) was added to a solution of cyclam (0.100 g, 0.50 mmol) and NaHCO.sub.3 (0.088 g, 1.05 mmol) in CH.sub.3CN (70 mL) and refluxed for 15 hrs. The white precipitate that formed was filtered off and the solvent was removed in vacuo. The residue was then purified with column chromatography (CH.sub.2Cl.sub.2: MeOH: NH.sub.4OH 10:1:0.1) to give the title compound as a slightly yellow oil which crystallised. The solid was recrystallised with toluene to yield clear crystals (0.126 g, 46%). δ.sub.H (CDCl.sub.3, 400 MHz): 9.01 (2 H, bs, —NH), 3.42 (2H, s, H-1′), 3.38 (2H, s, H-1′), 3.29 (2H, m, H-12), 3.17 (2H, m, H-9), 3.11 (2H, s, H-1′), 3.03 (2H, m, H-10), 2.74 (2H, t, J=5.6 Hz, CH.sub.2N), 2.70 (2H, m, CH.sub.2N), 2.63 (2H, t, J=5.6 Hz, CH.sub.2N), 2.59 (4H, m, CH.sub.2N), 2.03 (2H, bm, H-13), 1.66 (2H, m, H-6), 1.46 (9H, s, H-4′), 1.45 (9H, s, H-4′), 1.43 (9H, s, H-4′)

δ.sub.C (CDCl.sub.3, 100 MHz): 171.1 (C═O), 170.8 (C═O), 170.5 (C═O), 82.3 (C-3′), 81.6 (C-3′), 81.2 (0-3′), 55.8 (C-1′), 55.7 (C-1′), 55.3 (C-1′), 53.8 (CH.sub.2N), 52.0 (C-12), 51.2 (C-9), 50.5 (CH.sub.2N), 49.2 (CH.sub.2N), 48.5 (C-10), 47.6 (CH.sub.2N), 46.7 (CH.sub.2N), 28.2 (C-4′), 23.3 (C-6), 22.5 (C-13)
1-(10-(tetra-O-benzoyl-β-D-glucopyranos-1-yl)decyl)-4,8,11-Tris-(tert-butoxycarbonylmethyl)-1,4,8,11-tetraazacyclotetradecane (35)

##STR00038##

[0170] Bromide 4 (0.590 g, 0.72 mmol) was dissolved in anh. MeCN (10 mL) and added to a solution of cyclam 34 (0.327 g, 0.60 mmol) and NaHCO.sub.3 (0.152 g, 1.80 mmol) in MeCN (50 mL). The reaction was refluxed at 80° C. for 48 hrs after which H.sub.2O (1 mL) was added and the reaction again refluxed for 48 hrs. The solvent was evaporated under vacuum and the crude material redissolved in DCM (20 mL) and water (20 mL). The aqueous phase was extracted with DCM (3×20 mL) and the organic phase dried, filtered and concentrated to yield a crude oil which was purified with column chromatography using DCM:MeOH (9.5: 0.5) as the mobile phase. The title product was obtained as a clear oil (0.370g, 48%). 8.sub.H (CDCl.sub.3, 300 MHz): 8.02-7.81 (8H, m, ArH), 7.55-7.25 (12H, m, ArH), 5.89 (1H, t, J=9.6 Hz, H-3″), 5.66 (1H, t, J=9.6 Hz, H-4″), 5.51 (1H, dd, J=7.8, 9.7 Hz, H-2″), 4.83 (1H, d, J=7.8 Hz, H-1″), 4.63 (1H, dd, J=3.4, 12.1 Hz, H-6a″), 4.50 (1H, dd, J=5.2, 12.1 Hz, H-6b″), 4.15 (1H, m, H-5″), 3.90 (1H, dt, J=6.2, 9.7 Hz, H-24a), 3.53 (1H, dt, J=6.2, 9.7 Hz, H-24b), 3.53 (2H, m, CH.sub.2N), 3.32-3.22 (8H, m, CH.sub.2N, 3×H-1′), 3.12-3.06 (4H, m, H-15, CH.sub.2N), 2.72-2.60 (10H, m, 5×CH.sub.2N), 1.99 (2H, m, H-13), 1.78 (2H, m, H-16), 1.61 (2H, m, H-6), 1.52 (2H, m, H-23), 1.45 (27H, s, H-4′), 1.30-1.05 (12H, m, CH.sub.2-alk)

δ.sub.C (CDCl.sub.3, 100 MHz): 170.7 (C-2′), 170.5 (C-2′), 170.7 (C-2′), [166.1, 165.8, 165.2, 165.0 (C=0)], [133.3, 133.1, 133.1, 133.0, 129.8 (x2), 129.7 (x4), 129.6 (x2), 129.4 (x2), 128.9 (x2), 128.4 (x4), 128.3 (x2), 128.2 (x2)(ArC)], 101.3 (C-1″), 81.4 (C-3′), 81.4 (C-3′), 81.0 (C-3′), 73.0 (C-3″), 72.1 (C-2″), 72.0 (C-5″), 70.3 (C-24), 69.9 (C-4″), 63.3 (C-6″), 55.7 (C-1′), 55.7 (C-1′), 55.2 (C-1′), 53.2 (C-15), [52.1, 51.9, 51.8, 51.0, 50.7, 50.2, 50.0, 49.6 (CH.sub.2N)], [29.3, 29.3, 29.2, 29.1, 29.0 (CH.sub.2)], 28.2 (x3)(C-4′), 26.9 (C-22), 25.7 (C-17), 25.4 (C-6), 23.4 (C-16), 22.5 (C-13)
1-(10-(β-D-glucopyranos-1-yl)decyl)-4,8,11-Tris-(acetic acid) -1,4,8,11-tetraazacyclotetra-decane trifluoroacetate salt (36)

##STR00039##

[0171] Sodium methoxide (0.20 mL, 25% solution) was added to a solution of cyclam 35 (0.110 g, 0.09 mmol) in anh. MeOH (4.0 mL) and stirred for 1 hr. The reaction was quenched with 0.25 M HCl (10 mL) and extracted with DCM (3×10 mL). The aqueous phase was basified to pH 10-11 with 2 M NaOH and again extracted with DCM (3×10 mL). The organic phase was dried, filtered and concentrated to yield a crude oil. This oil was then dissolved in DCM (2 mL) and trifluoroacetic acid (2.0 mL) added to the solution which was then stirred overnight. All solvent was then evaporated and the remaining oil redissolved in water (5 mL) and extracted with CHCl.sub.3 (3×3 mL). The aqueous phase was concentrated down and further dried under high vacuum to yield the TFA-salt of the title compound as a glassy solid (0.032 g, 36%)

δ.sub.H (D.sub.2O, 400 MHz): 4.32 (1H, d, J=7.8 Hz, H-1″), 3.93 (2H, s, H-1′), 3.79 (2H, m, H-6″a/24a), 3.61-3.51 (6H, m, H-6″b/24b/1″(x2)), 3.40-3.25 (11H, m, H-3″/4″/5″/NCH.sub.2(x4)), 3.16-3.08 (7H, H-2″/15/NCH.sub.2(x2)), 2.90 (4H, bm, —NCH.sub.2(x2)), 1.93 (4H, m, H-6/13), 1.59 (2H, m, H-16), 1.50 (2H, m, H-23), 1.25-1.15 (12H, m, CH.sub.2-alk(x6))
δ.sub.C (D20, 100 MHz): 173.8 (C-2′), 173.7 (C-2′), 169.6 (C-2′), 102.1 (C-1″), 75.8 (C-3″), 75.8 (C-5″), 73.1 (C-2″), 70.6 (C-24), 69.9 (C-4″), 60.8 (C-6″), 54.7 (C-1′), 54.5 (C-1′), 54.4 (C-1′), 53.9 (C-15), [52.9, 52.5, 51.6, 51.5, 50.6, 50.0, 49.6, 48.9 (CH.sub.2N)], [28.7, 28.4, 28.3, 28.3, 28.0, 25.6, 24.9 (CH.sub.2)], 22.5 (C-16), [21.6, 21.0 (C-6/13)]
Procedure for Labelling Cyclam X with .sup.103Pd

##STR00040##

[0172] .sup.103Pd(NH.sub.3).sub.4Cl.sub.2 was dissolved in water (150 uL)(approx. pH 5) and the pH adjusted to 10-11 with 5M NaOH (4 uL). Cyclam 36 (50 uL 0.450 mg) was added to the .sup.103Pd(NH.sub.3).sub.4C1.sub.2 solution in and the mixture heated at 80° C. for 30 min. The reaction solution (20 uL) was then analysed by injection into a HPLC fitted with a Agilent Zorbax Extend C-18 Sum, 4.6×250 mm column and a Raytest Gabi Star Gamma detector and analysed with a mobile phase system of A: 0.01 M ammonium acetate pH 9.5, B: methanol run under a gradient elution (0 min A:B=95:5; 2 min A:B=80:20; 4 min A:B=50:50; 10 min A:B=0:100) with a flow rate of 0.8 mL/min. Free metal eluted at retention time of 1.35 min while labelled product had a retention time of 6.6 min.

Example 8

Radiolabelling—Radiolabelling of a Cyclam Pro-Conjugate 27 for Proof of Principle of .SUP.103.Pd Co-Ordination

[0173] ##STR00041##

[0174] .sup.103Pd(NH.sub.3).sub.4Cl.sub.2 fraction (32 MBq) obtained from .sup.103Pd purification on anion exchange column was dissolved in milliQ-water (200 uL)(approx. pH 4). Cyclam27 (100 uL, 1.0 mg) was added to the .sup.103Pd(NH.sub.3).sub.4Cl.sub.2 solution (100 uL) in a vial and the mixture heated at 85° C. for 30 min. The reaction solution (20 uL) was then analysed by injection into a HPLC fitted with a Agilent Zorbax Extend C-18 5 um, 4.6×250 mm column and a Raytest Gabi Star Gamma detector and analysed with a mobile phase system of A: 0.01 M ammonium acetate pH 9.5, B: methanol run under a gradient elution (0 min A:B=95:5; 2 min A:B=80:20; 4 min A:B=50:50; 10 min A:B=0:100) with a flow rate of 0.8 mL/min. Free metal eluted at retention time of 1.35 min while labelled product had a retention time of 7.9 min.

Example 9

Conversion of an Amine to a Maleimide Moiety

N-(Methoxycarbonyl)maleimide (37)

[0175] ##STR00042##

[0176] Methyl chloroformate (0.87 mL, 11.3 mmol) was added slowly to a solution of maleimide (1.00 g, 10.3 mmol) and N-methyl morpholine (1.24 mL, 11.3 mmol) in EtOAc (80 mL) at 0° C. and stirred for 1 hr. The precipitate was separated out through filtration through a celite pad and the filtrate concentrated in vacuo. It was attempted to recrystallise the crude oil with hexane: CH.sub.2Cl.sub.2 but no crystallisation occurred. The crude product was redissolved in EtOAc (100 mL), adsorbed onto silica and purified using column chromatography (Hex: EtOAc 6:4) to yield a white solid (1.07 g, 67%).

δ.sub.H (CDCl.sub.3, 300 MHz): 6.83 (2H, s, CH-3/4), 3.94 (3H, s, CH.sub.3-2′)
δ.sub.C (CDCl.sub.3, 100 MHz): 165.6 (C-2/5), 148.1 (C-1′), 132.3 (C-3/4), 54.2 (C-2′)
N-10-(β-D-glucopyranos-1-yl)decyl-maleimide (38)

##STR00043##

[0177] N-(methoxycarbonyl) maleimide(37) (0.014 g, 0.09 mmol) was added at RT to a stirring solution of (10-Aminodecyl) β-D-glucopyranoside(0.015 g, 0.04 mmol) in dioxane (0.75 mL) and sat. NaHCO.sub.3 (.sub.aq) (1.5 mL). The reaction was stirred for 1 hr after which the reaction was quenched with EtOAc (10 mL) and water (10 mL). The organic phase was separated out and the aq. phase extracted with EtOAc (3×10 mL). The organic layers were combined, dried, filtered and the solvent evaporated under vacuum. The residue was adsorbed onto silica and purified with column chromatography (CH.sub.2Cl.sub.2: MeOH 9:1). The title compound was obtained as a clear oil (0.011 g, 60%) R.sub.f=0.15 (CH.sub.2Cl.sub.2: MeOH 9:1)

δ.sub.H (CD.sub.3OD, 300 MHz): 6.79 (2H, s, 3′/4′), 4.24 (1H, d, J=7.5 Hz, H-1″), 3.93-3.84 (2H, m, H-6″a/H-10a), 3.69-3.64 (1H, m, H-6″b), 3.57-3.50 (1H, dt, J=6.6, 9.6 Hz, H-10b), 3.48 (2H, t, J=6.9 Hz, H-1), 3.38-3.22 (3H, m, H-3″/4″/5″), 3.19-3.13 (1H, m, H-2″), 1.64-1.54 (4H, m, H-2/9), 1.42-1.28 (12H, m, AlkCH.sub.2). δ.sub.C (CD.sub.3OD, 100 MHz): 172.6 (C-2′/5′), 135.3 (C-3′/4′), 104.4 (C-1″), 78.2 (C-3″), 77.9 (C-2″), 75.1 (C-5″), 71.7 (C-10), 70.9 (C-4″), 62.8 (C-6″), 38.5 (C-1), 30.8 (C-2), 30.5 (x3), 30.1, 29.4, 27.7, 27.0 (CH.sub.2Alk)

Example 10

[0178] Insertion of Maleimide Moiety into Pro-Conjugate for Connection to Albumin Carrier
Synthesis of non-cleavable linker pro-conjugate with maleimide moiety
1-[10-(β,D-glucopyranos-1-yl)]-4,11-(diacetic acid)-8-[10-maleimidodecyl]-1,4,8,11-tetraaza cyclotetradecane (39)

##STR00044##

[0179] N-(methoxycarbonyl) maleimide(37) (0.008 g, 0.05 mmol) is added at RT to a stirring solution of cyclam 27 (0.015 g, 0.02 mmol) in dioxane (0.75 mL) and sat. NaHCO.sub.3 (aq) (1.5 mL). The reaction is stirred for 1 hr after which the reaction is quenched with 0.25 M HCl (5 mL). The aqueous phase is extracted with EtOH:CHCl.sub.3(2:1)(5×5 mL). The organic layers are combined, dried, filtered and the solvent evaporated under vacuum to yield the product as an oil.

Synthesis of Cleavable Linker Pro-Conjugate with Maleimide Moiety
1-[10-(β,D-glucopyranos-1-yl)]-4,11-(diacetic acid)-8-[3-((maleimidodecyl)disulfanyl)propyl]-1,4,8,11-tetraazacyclotetradecane (40)

##STR00045##

[0180] N-(methoxycarbonyl) maleimide(37) (0.008 g, 0.05 mmol) is added at RT to a stirring solution of cyclam 32(0.015 g, 0.02 mmol) in dioxane (0.75 mL) and sat. NaHCO.sub.3 (aq) (1.5 mL).The reaction is stirred for 1 hr after which the reaction is quenched with 0.25 M HCl (5 mL). The aqueous phase is extracted with EtOH:CHCl.sub.3(2:1)(5×5 mL). The organic layers are combined, dried, filtered and the solvent evaporated under vacuum to yield the product as an oil.

1-[10-(β,D-glucopyranos-1-yl)]-4,11-(diacetic acid)-8-[3-((maleimidopropyl-1H-1,2,3-triazol-4-yl)propyl))disulfanyl)propyl]-1, 4,8,11-tetraazacyclotetradecane (41)

##STR00046##

[0181] 3-Azidopropyl-1-maleimide (0.002 g, 0.02 mmol) is added at RT to a stirring solution of diisopropyethylamine (0.0013 g, 0.01 mmol),cyclam 33 (0.015 g, 0.02 mmol) and catalytic copper iodide (0.2 eq) in DMF (1.0 mL) The reaction is stirred for 2 hr after which the reaction was quenched with 0.25 M HCl (1 mL). All solvent is evaporated under vacuum and the residue redissolved in 0.25 M HCl (2 ml) followed by extraction of the product with EtOH:CHCl.sub.3(2:1)(5×3 mL). The organic extracts are combined, dried, filtered and concentrated to yield the title compound.

Example 11

Attachment of Maleimido-Functionalised Compounds to Albumin

[0182] Reaction of albumin with maleimido-glycoside 38

[0183] Control solutions of BSA and 38 were made by dissolving BSA (0.016 g, 0.24 umols) and 38 (0.001 g, 2.4 umols) each in 1×PBS (1 mL). 20 uL of each control solution was analysed by HPLCunder a gradient elution with A: CH.sub.3CN (0.1% TFA) and B: H.sub.2O (0.1% TFA), 0-60% A over 30 min.HPLC analysis was done on an Agilent 1220 Infinity LC with an Agilent Zorbax Eclipse Plus C-18 (4.6×150 mm 5 um) column. The control solutions were then heated at 37° C. for 24 hrs and re-analysed by HPLC using the same method.

[0184] Fresh solutions of BSA and 38 were prepared as above and reacted together in an Eppendorf vial by heating at 37° C. After 5 min, a sample of the solution was analysed by HPLC using the same method as above.

Reaction of albumin with maleimido-pro-conjugate 39

[0185] Control solutions of BSA and 39 are made by dissolving BSA (0.007 g, 0.11 umols) and 39 (0.001 g, 1.1 umols) each in 1×PBS (1 mL). 20 uL of each control solution was analysed by HPLCunder a gradient elution with A: CH.sub.3CN (0.1% TFA) and B: H.sub.2O (0.1% TFA), 0-60% A over 30 min. HPLC analysis is done on an Agilent 1220 Infinity LC with an Agilent Zorbax Eclipse Plus C-18 (4.6×150 mm 5 um) column. The control solutions are then heated at 37° C. for 24 hrs and re-analysed by HPLC using the same method.

[0186] Fresh solutions of BSA and 39 are prepared as above and reacted together in an Eppendorf vial by heating at 37° C. After 5 min, a sample of the solution was analysed by HPLC using the same method as above.