Multimodal contrast and radiopharmaceutical agent for an imaging and a targeted therapy guided by imaging

Abstract

A multimodal contrast and radiopharmaceutical agent for an imaging and a targeted therapy guided by imaging.

Claims

1. Functionalized dendritic nanoprobes of the following formula (I) ##STR00021## wherein: for generation 1, R.sub.4 and R.sub.6 represent independently from each other a chain composed of oligoethyleneglycol patterns, at least one of said oligoethyleneglycol chains being functionalized at its extremity by a group chosen among a fluorophore, or a biocompatible dye, and R.sub.5 represents an hydrogen atom or a chain composed of oligoethyleneglycol patterns, said chain being optionally functionalized at its extremity by a group chosen among or a biocompatible dye, for higher generations, R.sub.5 represents an hydrogen atom and R.sub.4 and R.sub.6 represent a dendritic structure (D)m comprising at least one ether of benzyl alcohol, said benzyl being substituted either at positions 3, 4, and 5, or at positions 3 and 5, by chains composed of oligoethyleneglycol patterns, at least one of said oligoethyleneglycol chains being functionalized at its extremity by a group chosen among a fluorophore, or a biocompatible dye, and m=1, 2 or 4, X represents a group of the following formula (II): ##STR00022## wherein p is from 3 to 6 and wherein the mean diameter of the nanoprobe is from 2 to 60 nm.

2. The nanoprobes according to claim 1, wherein the group of formula (II) is complexed to a ligand or a radioelement to give a group of formula (III): ##STR00023## wherein ##STR00024## represents: a metallic ion, or a gamma radiation emitter radio-element or a positon emitter radio-element, or an alpha or beta negative radiation emitter radio-element.

3. The nanoprobes according to claim 1, wherein the formula (I) is selected from the following formulae: ##STR00025## ##STR00026## wherein R.sub.1 represents the group X of formula (II) as previously defined, n is an integer from 1 to 10, r is an integer from 1 to 20, r being n, and R.sub.a and R.sub.b independently represents a group selected from: a fluorophore, or a biocompatible dye bearing at least one group SO.sub.3R.sub.3, wherein R.sub.3 represents an hydrogen, sodium or calcium atom and eventually one or more groups chosen among OH and CO.sub.2H, or a linear or branched alkyl.

4. The nanoprobes according to claim 3, wherein the structure of formula (I) has the following formulae: ##STR00027## ##STR00028## ##STR00029## ##STR00030##

5. The nanoprobes according to claim 4, further comprising a complexed .sup.99mtechnetium.

6. The nanoprobes according to claim 1, in a form suitable for its use as a medicament, or for detecting and/or treating a cancer cell or tissue or organ.

7. The nanoprobes according to claim 6, wherein said cancer is brain cancer.

8. The nanoprobes according to claim 7, wherein said cancer is breast cancer and the organ is the sentinel node.

9. Pharmaceutical or diagnostic compositions comprising the nanoprobes according to claim 1.

10. The nanoprobes according to claim 2, wherein the metallic ion is gadolinium or manganese, and/or the gamma radiation emitter radio-element or positon emitter radio-element is .sup.99mtechnetium, .sup.64copper, .sup.(67, 68)gallium, or .sup.124iodine, and/or the alpha or beta negative radiation emitter radioelement is .sup.177lutetium, .sup.90yttrium, .sup.166holmium or .sup.186rhenium.

11. The nanoprobes according to claim 1, wherein the mean diameter of the nanoprobe is from 15 to 50 nm.

12. The nanoprobes according to claim 1, wherein the mean diameter of the nanoprobe is from 20 to 35 nm.

Description

DESCRIPTION OF THE FIGURES

(1) FIGS. 1A to 1E present the scintigraphic imaging (Single Photon Emission Computed Tomography SPECT) obtained with the nanoprobe (Ia1a) after intraveinous injection in rats at a dose of 20 MBq of 99mTc-Dopadendron (Ia1a).

(2) No adverse side-effect was observed after intraveinous (IV) injection in rats and mice.

(3) Planar dynamic acquisition acquired just after intra-venous injection in right saphene vein:

(4) FIG. A: 10 sec image at 1 min post-injection;

(5) FIGS. B and C30 sec

(6) Images respectively 10 and 20 minutes after injection of 20 MBq of 99mTc-Dopadendron

(7) (K=kidney activity, SI=injection site, C=cardiac activity, L=liver activity);

(8) FIGS. D and E: SPECT-CT images acquired at 30 min to 1 H30 after injection

(9) (FIG. D=posterior projection, FIG. E=left oblique posterior projection).

(10) FIGS. 1A to 1E show a rapid vascular dispersion, a low liver uptake, a rapid and intense renal activity. Vascular activity persisted 20 minutes after IV injection. SPECT revealed a digestive uptake, such as observed in human after injection of 18F-Dopa. One hour after injection, liver and vascular activity have disappeared, and only renal uptake was intense. No reticuloendothelial system (RES) uptake was observed.

(11) FIG. 2 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of allyl-protected compound 3.

(12) FIG. 3 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of allyl-protected compound 4.

(13) FIG. 4 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of compound 6.

(14) FIG. 5 presents the .sup.13C NMR (75 Mhz, CDCl.sub.3) of compound 6.

(15) FIG. 6 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of compound 9.

(16) FIG. 7 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of compound 11.

(17) FIG. 8 presents the .sup.1H NMR (300 Mhz, CDCl.sub.3) of compound 12.

(18) FIG. 9 presents the .sup.13C NMR (75 Mhz, CDCl.sub.3) of compound 12.

EXAMPLES

Example 1

Synthesis of Nanoprobes Ia1a Wherein n=2 Complexed with 99mTc

Example 1.1

Catechol Synthesis

(19) Catechol has been synthesized according to scheme I:

(20) ##STR00018##

(21) Materials and Synthesis:

(22) Reagents and solvents were purchased reagent grade and used without further purification.

(23) Compounds 5.sup.1, 10.sup.2 and 13.sup.3 were prepared according to the literature (respectively: .sup.1: D. Imbert, F. Thomas, P. Bare!, G. Serratrice, D. Gaude, J.-L. Pierre, J.-P. Laulhre, New J. Chem. 2000, 24, 281-288; .sup.2: M. Ou, X. L. Wang, R. Xu, C. W. Chang, D. A. Bull, S. W. Kim, Bioconj. Chem., 2008, 19, 626-633; .sup.3: A. Bertin, 1. Steibel, A.-I. Michou-Gallani, J.-L. Gallani, D. Felder-Flesch, Bioconj. Chem. 20, 760-767).

(24) All reactions were performed in standard glassware under Ar and solvents were, if necessary, purified by standard procedures prior to use. Evaporation and concentration were done at water-aspirator pressure and drying in vacuo at 10.sup.2 Torr (1.33 Pa). Column chromatography: silica gel 60 (230-400 mesh, 0.040-0.063 mm) from E. Merck. NMR spectra: Bruker AM-300 (300 MHz); solvent peaks as reference; in ppm.

(25) Compound 3:

(26) a solution of 2,3-dihydroxybenzoc acid (10.00 g, 64.89 mmol) and K.sub.2CO.sub.3 (33.20 g, 240.07 mmol) in acetonitrile (140 mL) was heated at 80 C. for 1 h. The reaction mixture was then cooled to room temperature and a solution of allyl bromide (20.80 mL, 240.07 mmol) in acetonitrile (80 mL) was added drop wise. The resulting mixture was heated 17 h at 70 C. and filtered. The filtrate was evaporated to dryness and diluted in EtOH (100 mL). After adding a solution of NaOH (7.40 g, 184.92 mmol) in 12 mL water, the reaction mixture was refluxed for 23 h and evaporated to dryness. The so-obtained residue was dissolved in CH.sub.2Cl.sub.2 (100 mL) and 150 mL of HCl 1 N were added. The aqueous sub phase was extracted two times by using CH.sub.2Cl.sub.2 (200 mL) and the organic phases put together, washed twice with water (200 mL), dried (MgSO.sub.4), filtered and evaporated. After recristallisation in a mixture hexane/ether (50 mL/50 mL), compound 3 (9.88 g, 42.18 mmol) was obtained with 65% yield. White powder. M.p 138 C. .sup.1H NMR (CDCl.sub.3): 4.62 (d, .sup.3J=5 Hz, 2H), 4.82 (d, .sup.3J=6 Hz, 2H), 5.32-5.50 (m, 4H), 6.01-6.15 (m, 2H), 7.16 (m, 2H), 7.75 (dd, .sup.3J=6 Hz, IH); C.sub.13H.sub.14O.sub.4: calc. C, 66.66; H, 6.02; O, 27.32. found C, 66.58; H, 6.02; O, 27.40.

(27) Compound 4:

(28) 0.51 mL (5.977 mmol) of trifluorotriazine were added to a stirred solution of 3 (2.00 g, 8.54 mmol) in dry CH.sub.2Cl.sub.2 (100 mL) cooled to 0 C. The mixture was stirred 10 min before addition of an anhydrous CH.sub.2Cl.sub.2 (30 mL) solution of pyridine (0.76 mL, 9.39 mmol). After 17 h at room temperature, the mixture was washed with cold water (275 mL) and NaCl saturated water (75 mL). The organic sub phase was then dried (MgSO.sub.4) filtered and evaporated. Crude compound 4 was obtained as brown oil (1.82 g, 7.69 mmol) with 90% yield and used in next step without further purification. .sup.1H NMR (CDCl.sub.3): 4.60-4.66 (m, 4H), 5.23-5.48 (m, 4H), 6.00-6.19 (m, 2H), 7.09-7.22 (m, 2H), 7.49 (d, .sup.3J=9 Hz, IR); .sup.13C NMR (CDCl.sub.3): 70.03, 75.02, 117.95, 118.42, 120.32, 123.99, 124.10, 132.46, 133.55, 150.50, 152.76, 157.46.

(29) Compound 6:

(30) Compound 4 (4.96 g, 21.00 mmol) in freshly distilled CH.sub.2Cl.sub.2 (100 mL) was added dropwise over 90 min to a stirred solution of N,N-diisopropylethylamine (4.52 mL, 27.36 mmol) and 5 (2.11 g, 6.36 mmol) in freshly distilled CH.sub.2Cl.sub.2 (100 mL) kept under nitrogen. After 60 h stirring at room temperature, the crude mixture was filtered, washed with water, dried (MgS04), filtered and then evaporated to dryness. Column chromatography (SiO.sub.2, CH.sub.2Cl.sub.2/1% MeOH) allowed to obtain compound 6 (6.05 g, 6.17 mmol) in 97% yield. Pale yellow oil. .sup.1H NMR (CDCl.sub.3): 0.01 (s, 6H), 0.82 (s, 9H), 1.25-1.48 (m, 12H), 3.29 (s, 2H), 3.34-3.38 (m, 6H), 4.57 (d, 12H), 5.24-5.46 (m, 12H), 6.00-6.12 (m, 6H), 6.99 (d, 3 J=13 Hz, 3H), 7.03 (d, .sup.3J=9.7 Hz, 3H), 7.68 (d, .sup.3J=9 Hz, 3H), 8.02 (t, 3H); .sup.13C NMR (CDCl.sub.3): 5.67, 18.06, 23.35, 25.76, 31.48, 39.38, 40.54, 66.51, 69.77, 74.59, 116.75, 117.67, 118.76, 123.17, 124.23, 127.50, 132.80, 133.19, 146.24, 151.48, 164.98; C.sub.56H.sub.77N.sub.3O.sub.10Si. H.sub.2O: calc. C, 68.09; H, 7.90; N, 4.26. found C, 68.23; H, 7.83; N, 4.12.

(31) Compound 7:

(32) Compound 6 (735 mg, 0.75 mmol) was dissolved in 10 mL THF at 0 C. Tetra-n-butylammonium fluoride (2.21 mmol) was slowly added and the reaction mixture was heated at reflux for 3 h. The solution was evaporated and the so-obtained residue was dissolved in CH.sub.2Cl.sub.2 (100 mL), washed with water, dried (MgSO.sub.4), filtered and evaporated. Column chromatography (SiO.sub.2, CH.sub.2Cl.sub.2/3% MeOH) afforded 7 (610 mg, 0.71 mmol) in 94% yield. Yellow oil. .sup.1H NMR (CDCl.sub.3): 1:1.27-1.63 (m, 12H), 3.39 (m, 8H), 4.58 (d, 12H), 5.26-5.46 (m, 12H), 6.00-6.12 (m, 6H), 7.02 (d, .sup.3J=8. Hz, 3H), 7.12 (t, .sup.3J=8 Hz, 3H), 7.68 (d, .sup.3J=9 Hz, 3H), 8.08 (t, .sup.3J=5 Hz, 3H), .sup.13C NMR (CDCl.sub.3): 23.27, 31.14, 39.41, 40.43, 66.39, 69.76, 74.63, 116.78, 117.73, 118.87, 123.13, 124.37, 127.39, 132.78, 133.21, 146.24, 151.48, 158.25, 165.18; C.sub.50H.sub.63N.sub.3O.sub.10: calc. C, 69.34; H, 7.33. found C, 69.04; H, 7.78.

(33) Compound 8:

(34) A solution of anhydrous dimethyl sulfoxide (144 mL, 2.03 mmol) in 300 L dry dichloromethane was added to a solution of oxalyl chloride (89 L, 1.02 mmol) in freshly distilled dichloromethane (1 mL) kept at 60 C. The mixture was stirred for 15 min and then a solution of 7 (0.80 g, 0.92 mmol) in dry dichloromethane (3 mL) was added within a 10 min period; the solution obtained was stirred for an additional 1 h and the reaction mixture was allowed to warm to 30 C. N, N-diisopropylethylamine (10 eq.) was added and the reaction mixture was allowed to warm to room temperature. Water (50 mL) and dichloromethane (50 mL) were added and the organic layer was washed with brine, dried (MgS04), filtered and evaporated to dryness. Compound 8 was used in the next step without further purification. Colourless oil. .sup.1H NMR (CDC.sub.3): 1.26-1.58 (m, 12H), 3.40 (m, 6H), 4.58 (d, 12H), 5.26-5.47 (m, 12H), 6.02-6.11 (m, 6H), 7.01 (d, .sup.3J=9 Hz, 3H), 7.11 (t, .sup.3J=8 Hz, 3H), 7.68 (d, .sup.3J=7 Hz, 3H), 8.08 (t, 3H), 9.42 (s, IH); .sup.13C NMR (CDCl.sub.3): 23.72, 29.30, 39.93, 51.29, 69.75, 74.49, 116.86, 117.71, 118.79, 123.09, 124.27, 127.20, 132.74, 133.15, 146.25, 151.46, 165.13, 206.59.

(35) Compound 9:

(36) Sulfamic acid (466 mg, 4.80 mmol) and then sodium chlorite (434 mg, 4.80 mmol) were added to a stirred solution of 8 (3.19 g, 3.69 mmol) in a mixture THF/water (1/1). After 13 h stirring at room temperature, 200 mL CH.sub.2Cl.sub.2 and 200 mL water were added and the resulting organic layer was washed with brine, dried (MgSO.sub.4), filtered and evaporated to dryness. Column chromatography (SiO.sub.2, CH.sub.2Cl.sub.2/3% MeOH) afforded 9 (0.47 g, 0.53 mmol) with 58% yield over the two steps. Pale yellow oil. .sup.1H NMR (CDCl.sub.3): 1.53-1.66 (m, 12H), 3.40 (d, 6H), 4.57 (d, 12H), 5.24-5.45 (m, 12H), 5.99-6.12 (m, 6H), 6.98 (d, .sup.3J=9 Hz, 3H), 7.09 (t, .sup.3J=8 Hz, 3H), 7.68 (d, .sup.3J=9 Hz, 3H), 8.08 (t, 3H); .sup.13C NMR (CDCl.sub.3): 24.16, 31.92, 40.02, 48.06, 69.76, 74.64, 116.80, 117.72, 118.96, 123.12, 124.25, 127.34, 132.79, 133.16, 146.25, 151.48, 165.19, 178.96; C.sub.49H.sub.61N.sub.3O.sub.11. 1.5H.sub.2O: calc. C, 66.23; H, 7.06; N, 4.63; O, 22.08%. found C, 66.43; H, 6.83; N, 4.31; O, 22.43.

(37) Compound 11:

(38) A mixture of DCC (112 mg, 0.55 mmol) and DMAP (11 mg, 0.09 mmol) in dry CH.sub.2Cl.sub.2 (5 mL) and a solution of 109 (85 mg, 0.46 mmol) in dry CH.sub.2Cl.sub.2 (5 mL) were added to 9 (400 mg, 0.46 mmol) dissolved in freshly distilled dichloromethane (10 mL). A catalytic amount of HOBt was then added and the mixture obtained was stirred for 60 h at room temperature before filtration and evaporation to dryness. Column chromatography (SiO.sub.2, CH.sub.2Cl.sub.2/3% MeOH) afforded 11 (455 mg, 0.44 mmol) in 97% yield. Pale yellow oil. .sup.1H NMR (CDCl.sub.3): 1.41-1.55 (m, 25H), 3.07 (d, .sup.3J=5 Hz, 2H), 3.23 (d, .sup.3J=5 Hz, 2H), 3.37 (d, 6H), 4.56 (d, 12H), 5.24-5.45 (m, 12H), 5.98-6.11 (m, 6H), 6.98 (d, .sup.3J=9 Hz, 3H), 7.09 (t, .sup.3J=8 Hz, 3H), 7.66 (d, .sup.3J=9 Hz, 3H), 8.04 (t, .sup.3J=11 Hz, 3H), .sup.13C NMR (CDCl.sub.3). 24.28, 25.57, 26.83, 27.53, 28.40, 32.21, 40.00, 47.87, 69.77, 74.65, 116.82, 117.73, 118.96, 123.06, 124.25, 127.36, 132.77, 133.19, 146.26, 151.49, 156.14, 165.21, 175.60; C.sub.59H.sub.79N.sub.5O.sub.12. MeOH: calc. C, 66.58; H, 7.73; N, 6.47; O, 19.22. found C, 66.77; H, 7.75; N, 6.63; O, 18.85.

(39) Compound 12:

(40) TFA (68 mL, 0.89 mmol) was added to a solution of 11 (465 mg, 0.44 mmol) in dry CH.sub.2Cl.sub.2 (10 mL). After 6 h stirring at reflux, the reaction mixture was evaporated and filtered on a silica pad (SiO.sub.2, CH.sub.2Cl.sub.2/5% MeOH). Compound 12 (376 mg, 0.40 mmol) was obtained as a white foam in 90% yield. H NMR (CDCl.sub.3): 1.46-1.73 (m, 16H), 2.99 (d, 2H), 3.26-3.32 (m, 8H), 4.56 (d, 12H), 5.23-5.43 (m, 12H), 5.96-6.09 (m, 6H), 6.98 (d, .sup.3J=9 Hz, 3H), 7.06 (t, .sup.3J=8 Hz, 3H), 7.56 (d, 3H), 7.80 (br s, 2H), 8.34 (br s, 4H); .sup.13C NMR (CDCl.sub.3): 24.19, 24.60, 26.10, 31.84, 38.65, 39.87, 40.20, 47.94, 69.77, 74.67, 117.16, 117.76, 119.03, 122.64, 124.33, 126.76, 132.68, 133.06, 146.33, 151.52, 165.81, 176.71; C.sub.54H.sub.71N.sub.5O.sub.10. 2.5 CH.sub.2Cl.sub.2: calc. C, 58.59; H, 6.56; N, 5.99. found C, 58.25; H, 6.35; N, 5.67.

Example 1.2

N, N di-allyl-L-Dopa tris-catecholamide 17 and technetium complex thereof

(41) These compounds have been synthesized according to scheme II:

(42) ##STR00019## ##STR00020##
Compound 14:

(43) A solution of sodium hydroxide (0.11 g, 2.82 mmol) in 7.5 mL of water was added to a solution of 13 (1.60 g, 2.17 mmol) in dry THF (30 mL). The reaction mixture was stirred at room temperature for 24 h. After evaporation of THF, the resulting aqueous solution was acidified with concentrated HCl to pH=2 and extracted with CH.sub.2Cl.sub.2 (2150 mL). Drying of the combined organic layers over MgSO.sub.4 and evaporation of the solvent yielded the desired compound 14 as a yellow oil (1.49 g, 2.06 mmol) with 95% yield and used in next step without further purification. .sup.1H NMR (CDCl.sub.3): 1.19 (s, 18H), 3.45-3.53 (m, 4H), 3.55-3.70 (m, 24H), 3.72-3.90 (m, 6H), 4.15-4.30 (m, 6H), 7.28 (s, 2H), .sup.13C NMR (CDCl.sub.3): 27.29, 60.95, 61.40, 68.66, 69.46, 70.21, 70.35, 70.42, 70.45, 70.56, 70.95, 72.23, 72.27, 72.93, 109.24, 124.60, 142.43, 152.02, 168.97; C.sub.35H.sub.62O.sub.15: calc. C, 58.15; H, 8.65. found C, 58.20; H, 8.73.

(44) Compound 15:

(45) To a solution of amine 12 (0.58 g, 0.61 mmol) in dry CH.sub.2Cl.sub.2 (1 00 mL) were added EDCI (0.19 g, 0.98 mmol), DMAP (0.03 g, 0.24 mmol) and then slowly (within 2 h) a solution of carboxylic acid 14 (0.44 g, 0.61 mmol) in dry CH.sub.2Cl.sub.2 (50 mL). The obtained mixture was stirred for 24 h at room temperature and then was diluted with CH.sub.2Cl.sub.2 (50 mL). The organic phase was washed with saturated aqueous NH.sub.4Cl solution (2150 mL) and brine (1150 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo. Column chromatography (SiO.sub.2, AcOEt) afforded 15 (0.70 g, 0.42 mmol) as colorless oil in 70% yield. H NMR (CDCl.sub.3): 1.17 (s, 18H), 1.40-1.60 (m, 16H), 3.20-3.40 (m, 10H), 3.45-3.52 (m, 4H), 3.53-3.71 (m, 24H), 3.72-3.80 (m, 6H), 4.10-4.20 (m, 6H), 4.51-4.61 (m, 12H), 5.22-5.44 (m, 12H), 5.96-6.10 (m, 7H), 6.96-7.08 (m, 6H), 7.22 (s, 2H), 7.58 (d, .sup.3J=9 Hz, 3H), 7.88 (br s, 1H), 8.12 (br s, 3H); 13C NMR (CDCl.sub.3): 24.36, 27.00, 27.20, 27.40, 32.23, 38.20, 38.90, 40.02, 47.95, 53.38, 61.07, 61.55, 68.63, 69.57, 69.78, 70.30, 70.52, 70.53, 70.54, 71.13, 72.30, 72.40, 72.94, 74.65, 106.72, 116.92, 117.76, 118.95, 122.88, 124.29, 127.18, 130.01, 132.75, 133.20, 140.33, 146.30, 151.53, 152.14, 165.26, 166.73, 175.62; MS (MALDI-TOF) m/z: 1677.92 [M+Na].sup.+; C.sub.89H1.sub.31N.sub.5O.sub.24: calc. C, 64.59; H, 7.98; N, 4.23. found C, 65.50; H, 7.75; N, 4.87.

(46) Compound 16:

(47) To a solution of alcohol 15 (0.48 g, 0.29 mmol) in dry CH.sub.2Cl.sub.2 (35 mL) were added EDCI (0.09 g, 0.46 mmol), DMAP (0.015 g, 0.12 mmol) and then the allyl-protected L-DOPA (0.10 g, 0.29 mmol).

(48) The reaction mixture was refluxed for 72 h and then was diluted with CH.sub.2Cl.sub.2 (100 mL). The organic phase was washed with saturated aqueous NH.sub.4Cl solution (2150 mL) and brine (1150 mL), dried (MgSO.sub.4), filtered and evaporated to dryness. Column chromatography (SiO.sub.2, CH.sub.2Cl.sub.2/1% MeOH) afforded the compound 16 (0.30 g, 0.15 mmol) as colorless oil in 50% yield. .sup.1H NMR (CDCl.sub.3): 1.16 (s, 18H), 1.40-1.60 (m, 16H), 2.72-2.80 (m, IH), 2.92-3.07 (m, 3H), 3.20-3.40 (m, 12H), 3.42-150 (m, 4H), 3.51-3.78 (m, 30H), 4.06-4.15 (m, 6H), 4.21-4.31 (m, IH), 4.51-4.62 (m, 16H), 5.00-5.13 (m, 4H), 5.20-5.44 (m, 16H), 5.55-5.71 (m, 2H), 5.91-6.11 (m, 9H), 6.64-6.79 (m, 3H), 6.95-7.09 (m, 6H), 7.22 (s, 2H), 7.58 (d, .sup.3J=9 Hz, 3H), 7.84 (br s, IH), 8.12 (br s, 3H); .sup.13C NMR (CDCl.sub.3): 24.35, 27.03, 27.22, 27.43, 32.22, 35.38, 39.19, 39.78, 40.00, 47.93, 53.33, 61.06, 63.11, 63.62, 68.63, 68.90, 69.58, 69.76, 69.86, 70.02, 70.32, 70.53, 70.61, 71.12, 72.33, 72.88, 74.63, 106.79, 114.14, 115.60, 116.91, 116.98, 117.28, 117.73, 118.92, 121.89, 122.88, 124.26, 127.18, 130.00, 131.67, 132.74, 133.19, 133.54, 133.65, 136.36, 140.61, 146.29, 146.94, 148.16, 151.51, 152.19, 165.24, 166.73, 172.32, 175.60; MS (MALDI-TOF) m/z: 1995.00 [M+H].sup.+, 2016.86 [M+Na].sup.+; C.sub.110H1.sub.56N.sub.6O.sub.27: calc. C, 66.24; H, 7.88; N, 4.21. found C, 65.90; H, 7.78; N, 4.17.
Compound 17:

(49) To a solution of compound 16 (0.057 g, 0.028 mmol) in dry THF (4 mL) was added, under Ar, Pd(PPh.sub.3).sub.4 (0.005 g, 0.004 mmol). The reaction mixture was stirred for 5 minutes and then NaBH.sub.4 (0.008 g, 0.224 mmol) was added. The resulting suspension was stirred at room temperature, under Ar, for 3 h and then was quenched by MeOH (4 mL) addition. After evaporation of the solvent, the crude was purified by size exclusion chromatography (THF) to yield the compound 17 (0.038 g, 0.023 mmol) as brown solid in 81% yield. H NMR (CD.sub.3OD): 1.21 (s, 18H), 1.45-1.80 (m, 16H), 2.75-2.85 (m, 1H), 2.92-3.03 (m, 1H), 3.13-3.23 (m, 2H), 3.24-3.51 (m, 12H), 3.53-3.90 (m, 34H), 4.18-4.40 (m, 7H), 5.12-5.30 (m, 4H), 5.70-5.91 (m, 2H), 6.35-6.43 (m, 1H), 6.48-6.53 (m, 2H), 6.57-6.74 (m, 6H), 7.18 (d, .sup.3J=9 Hz, 3H), 7.38 (s, 2H); .sup.13C NMR (CDCl.sub.3): 25.40, 27.78, 28.00, 28.10, 30.11, 33.09, 40.20, 40.65, 40.85, 54.65, 61.98, 63.80, 64.00, 66.10, 69.35, 70.33, 71.05, 71.12, 71.27, 72.13, 73.65, 74.54, 107.10, 108.50, 110.30, 111.68, 114.27, 117.80, 118.06, 118.19, 119.35, 131.80, 137.49, 153.00, 153.20, 153.67, 153.84, 168.88, 170.40, 174.10, 178.80; MS (MALDI-TOF) m/z: 1674.86 [M+H].sup.+, 1679.12 [M+5H].sup.+; C.sub.86H.sub.124N.sub.6O.sub.27: calc. C, 61.71; H, 7.47; N, 5.02. found C, 63.50; H, 8.93; N, 4.11.

(50) Complexation of the compound 17 with technetium:

(51) The compound 17 is dissolved in water (2 mL, 1 mg/mL). A solution (250 microL) of stannous chloride (1 mg/mL, 1.3 mmol) in 0.1 M hydrochloric acid is first added to the above solution and then the 99mTc(VII)O4- solution (220 MBq mL/L). 25 mL of a 1 N aqueous solution of NaOH

(52) (25 mmol) and 200 mL of an aqueous solution of sodium ascorbate (150 mM) are also added in order to buffer the reaction mixture at pH 7 and to keep a low redox potential.

(53) The resulting mixture is then stirred at room temperature for 15 min The radiolabeling process with pertechnetate was efficient and reached 95% .sup.99mTc-complexation).