(99m)Tc complexes with activated ester functions; ligands comprising a 3,4-diamino-benzoate backbone.

Preformed (99m)Tc chelates with an activated ester function are useful for the gentle labeling of proteins (precomplexation route). In this context, new heterobifunctional ligands derived from 2,3,5,6-tetrafluorophenyl (TFP) 3,4-diamino-benzoates (OC1, OC3, OC4) were synthesized. Their corresponding (99m)Tc-complexation and protein-conjugation characteristics were elucidated and compared with the results previously reported using 2,3,5,6-tetrafluorophenyl N-(S-benzoylthioacetyl)glycylglycyl-p-aminobenzoate (OC2). The reaction temperatures and the reaction time markedly influenced complexation yields. Compared with OC2, the (99m)Tc-chelate formation with OC1 and OC4 was more effective, showing radiochemical yields of 60% and 70% within 20 min, respectively. Owing to steric hindrance, the complexation of OC3, however, did not exceed 10%. No-carrier-added (99m)Tc complexes were conjugated at pH 10 with the anti-EGF-receptor monoclonal antibody MAb425, resulting in labeling yields of 14% for (99m)Tc-OC1 and 7% for (99m)Tc-OC4 after incubating for 20 min at 30 degrees C. Increasing the temperature to 40 degrees C improved these results by 14% and 3%, respectively. As compared with (99m)Tc-OC2, which provides the chelating substituent at the 4-phenyl position only, the application of 3,4-phenyl substituents proved less appropriate for protein conjugation. However, the 3,4-diaminobenzoate backbone may be attractive for an alternative design of novel (99m)Tc N2S2 or N3S complexes, because they show excellent complexation characteristics.

Alkylating benzamides with melanoma cytotoxicity.

Radioiodinated N-(2-(diethylamino)ethyl)benzamides have recently been discovered as selective agents for melanotic melanoma and are used for scintigraphic imaging in nuclear medicine. Owing to the high binding capacity, benzamide derivatives conjugated with alkylating cytostatics were synthesized and tested for their potential for targeted drug delivery. Conjugates of chlorambucil with procainamide (1), diethylaminoethylamine (2) and 2-pyrrolidin-1-yl-ethylamine (3), as well as 4-(bis(2-chloroethyl)amino)- (6,7) and 4-(N,N-diethyltriazeno)-substituted (8-10) benzamides, were synthesized. Cell uptake studies with B16 melanoma cells revealed high uptake of radioiodinated 1 and 2, while radiolabelled chlorambucil was found to lack this characteristic. These results were confirmed by biodistribution studies in a mouse melanoma model. Viability measurements revealed that all chlorambucil-benzamide derivatives showed higher toxicity against B16 melanoma and SK-MEL-28 cells than did the parent chlorambucil itself, and that the triazene derivatives were more potent than dacarbazine, which is currently used as a standard cytostatic drug in melanoma therapy. Of all the compounds tested in this series, the triazenes 9 and 10 showed the most promising targeting effect. The toxicity of these compounds against hepatoma cells (MH3924A) and, to a lesser extent, against mouse fibroblast (NIH 3T3) and cervix carcinoma (HeLa) cells was also enhanced, but they were not as toxic as dacarbazine (HeLa). These findings support the concept of a selective, benzamide-mediated in vivo delivery of cytostatics in melanoma cells, leading to enhanced efficacy.

PET and SPECT for detection of tumor progression in irradiated low-grade astrocytoma: a receiver-operating-characteristic analysis.

UNLABELLED: Differentiation between tumor progression and radiation necrosis is one of the most difficult tasks in oncologic neuroradiology. Functional imaging of tumor metabolism can help with this task, but the choice of tracer is still controversial. This prospective study following up irradiated low-grade astrocytoma (LGA) was, to our knowledge, the first receiver-operating-characteristic (ROC) analysis that intraindividually evaluated the diagnostic performance of the SPECT tracers 3-[(123)I]iodo-alpha-methyl-L-tyrosine (IMT) and (99m)Tc(I)-hexakis(2-methoxyisobutylisonitrile) (MIBI) and the PET tracer (18)F-FDG. METHODS: We examined 17 patients, initially with histologically proven LGA and treated by stereotactic radiotherapy, who presented with new gadolinium-diethylenetriaminepentaacetic acid-enhancing lesions (n = 26) on MRI. At that time, MRI could not differentiate between progressive tumor and nonprogressive tumor. This MRI examination was closely followed by (18)F-FDG PET and by (99m)Tc-MIBI and (123)I-IMT SPECT. Lesions were classified as progressive tumor (n = 17) or nonprogressive tumor (n = 9) on the basis of prospective follow-up (through clinical examination, MRI, and proton MR spectroscopy) for 26.6 +/- 6.6 mo after PET or SPECT. RESULTS: (123)I-IMT yielded the best ROC characteristics and was the most accurate for classification, with an area under the ROC curve (A(z)) of 0.991. The A(z) of (18)F-FDG (0.947) was not significantly lower than that of (123)I-IMT. The difference in the A(z) of (99m)Tc-MIBI (0.713) from the A(z) of the other tracers used in our study was highly significant (P

Pharmacological properties of hydrophilic and lipophilic derivatives of octreotate.

Derivatives of somatostatin (SST) represent the most important peptides for receptor targeting in oncological applications. Whereas the pharmacophor in somatostatin receptor-affine substances has been thoroughly investigated, the influence of modifications at the N-terminal has not yet been systematically studied. In order to investigate the influence of hydrophilic versus lipophilic modifications at the N-terminal end, a series of homologous derivatives of Tyr3-octreotate modified with oligomers of ethylene glycol or fatty acids were synthesized. For this purpose, Tyr3-octreotate was assembled using solid phase peptide synthesis and the fatty acids or oligomers of ethylene glycol were conjugated to the N-terminal end. The oligomers of ethylene glycol were activated by 4-nitrophenylchloroformate to obtain carbamate-linked hydrophilic compounds. The receptor affinities of these compounds were determined by competition experiments with [125I]Tyr3-octreotide on rat cortex membranes. The hydrophilic derivatives and the short chain lipophilic derivatives revealed IC50 values between 0.66 +/- 0.02 nM and 2.16 +/- 0.31 nM respectively. After labeling with (125)I the organ distribution of selected derivatives was investigated in Lewis rats bearing the rat pancreatic tumor CA20948. All of the compounds showed high tumor uptake. The peptides conjugated to oligomers of ethylene glycol showed low uptake into the liver and kidneys. Increasing the length of the fatty acids resulted in a remarkable decrease in kidney uptake. In conclusion, the systematic modifications at the N-terminal result in a low effect on the receptor affinity but allow the modulation of the pharmacokinetic properties of octreotide derivatives.

Melanoma uptake of (99m)Tc complexes containing the N-(2-diethylaminoethyl)benzamide structural element.

On the basis of the avid uptake of radioiodinated benzamides by melanoma cells, (99m)Tc complexes containing the structural elements of N-(dialkylaminoalkyl)benzamide pharmacophores have been synthesized and evaluated in vitro and in vivo for melanoma uptake. One of the complexes Tc-12 containing the ligand 4-(S-benzoyl-2-thioacetyl-glycyl-glycylamido)-N-(2-diethylaminoethyl)benzamide (11) displayed the highest melanoma uptake. The 1-h melanoma uptake values and the corresponding blood counts indicate an interdependence of tumor uptake and bioavailability of the (99m)Tc complexes.

Detection of tumour progression in the follow-up of irradiated low-grade astrocytomas: comparison of 3-[123I]iodo-alpha-methyl- L-tyrosine and 99mTc-MIBI SPET.

Conventional MRI often fails to distinguish between progressive tumour and radiation injury, because both appear as mass lesions with unspecific Gd-DTPA enhancement. Furthermore, the sensitivity of FDG PET for the evaluation of malignant lesions in the brain is limited owing to high cortical uptake. The aim of this study was to assess the potential of alternative SPET tracers in the same group of patients. 35.2+/-20.1 months after stereotactic radiotherapy (59.3+/-4.2 Gy) of low-grade astrocytomas (median WHO II), 16 patients, presenting 25 Gd-DTPA-enhancing lesions on MRI, were examined by SPET. Lesions were classified as progressive tumour (PT, n=17) or non-PT (nPT, n=8) based on prospective follow-up (clinical examination, MRI, proton-MR spectroscopy) for 25.6+/-6.7 months after SPET. SPET scans were performed 15 and 60 min after injection of 694+/-67 MBq hexakis(2-methoxyisobutylisonitrile)(99m)Tc(I) (MIBI). 3-[(123)I]iodo-alpha-methyl- L-tyrosine (IMT) SPET was acquired 15 min after injection of 291+/-58 MBq IMT. Lesion-to-normal tissue ratios (l/n) for IMT (l/n(IMT)) and MIBI (l/n(MIBI)) were calculated using a reference region mirrored to the contralateral hemisphere. Using IMT, significantly higher ratios ( P<0.001) were found in PT (1.7+/-0.4) than in nPT (1.1+/-0.1). For MIBI, there was no statistically significant difference ( P=0.206) between PT (3.7+/-2.8) and nPT (1.8+/-1.8). Sensitivities for MIBI and IMT were 53% and 94%, and specificities 75% and 100%, respectively. Positive predictive values for MIBI and IMT respectively reached 80% and 100%, and negative predictive values were 46% and 90%. In conclusion, in contrast to MIBI, IMT showed almost no overlap between the PT and the nPT group. The sensitivity, specificity and predictive values of IMT SPET were obviously higher than those of MIBI SPET. IMT is considered to be a useful tracer for differentiating PT from nPT in the follow-up of irradiated low-grade astrocytomas.

Pretreatment evaluation of carcinomas of the hypopharynx and larynx with 18F-fluorodeoxyglucose, 123I-alpha-methyl-L-tyrosine and 99mTc-hexakis-2-methoxyisobutylisonitrile.

While fluorine-18 2-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) is helpful in the pretherapeutic evaluation of head and neck cancer, it is only available in selected centres. Therefore, single-photon emission tomography (SPET) tracers would be desirable if they were to demonstrate tumour uptake reliably. This multitracer study was performed to evaluate the pretherapeutic uptake of the SPET tracers iodine-123 alpha-methyl-L-tyrosine (IMT) and technetium-99m hexakis-2-methoxyisobutylisonitrile (99mTc-MIBI) in primary carcinomas of the hypopharynx and larynx and to compare the results with those of FDG PET. We examined 22 fasted patients (20 male, 2 female, mean age 60.5+/-10.2 years) with histologically confirmed carcinoma of the hypopharynx (n=9) or larynx (n=13), within 1 week before therapy. In 20 patients a cervical PET scan was acquired after intravenous injection of 232+/-43 MBq 18F-FDG. Data analysis was semiquantitative, being based on standardised uptake values (SUVs) obtained at 60-90 min after injection. After injection of 570+/-44 MBq 99mTc-MIBI, cervical SPET scans (high-resolution collimator, 64x64 matrix, 64 steps, 40 s each) were obtained in 19 patients, 15 and 60 min after tracer injection. Finally, 15 min after injection of 327+/-93 MBq 123I-IMT (medium-energy collimator, 64x64 matrix, 64 steps, 40 s each) SPET scans were acquired in 15 patients. All images were analysed visually and by calculating the tumour to nuchal muscle ratio. Eighteen of 20 (90%) carcinomas showed an increased glucose metabolism, with a mean SUV of 8.7 and a mean carcinoma to muscle ratio of 7.3. The IMT uptake was increased in 13 of 15 (87%) patients, who had a mean carcinoma to muscle ratio of 2.9. Only 13 of 19 (68%) carcinomas revealed pathological MIBI uptake, with a mean tumour to muscle ratio of 2.2 and no significant difference between early and late MIBI SPET images (P=0.23). In conclusion, in the diagnosis of primary carcinomas of the hypopharynx and larynx, IMT SPET achieved a detection rate comparable to that of FDG PET. IMT SPET was clearly superior to MIBI SPET in this population. A further evaluation of the specificity of IMT in a larger number of patients appears justified.