Development of phosphonate derivatives of gadolinium chelates for NMR imaging of calcified soft tissues.

We have synthesized several classes of gadolinium (Gd) complexes for use as NMR contrast agents in the detection of soft-tissue calcification. Class I was made up of strongly chelated GdDTPA complexes with one carboxylate arm coupled to a phosphonate-containing molecule through an amide link. Class II complexes were formed by Gd with several aminophosphonates and phosphono carboxylic acids. Class III were Gd complexes of weak chelates containing no phosphonate. The calcium-seeking ability of each complex was assessed by in vivo bone uptake. Tissue distribution in normal rats showed that only the complexes of GdDTPA modified with a diphosphonate group and GdEDTMP (EDTMP is ethylenediaminetetrakis(methylenephosponate] showed adequate bone localization at the concentrations required for NMR contrast enhancement (approximately 20% of a 100 mumol/kg dose).

p53 gene mutations in human astrocytic brain tumors including pilocytic astrocytomas.

Recent molecular biological studies have shown evidence for a distinct pathogenesis of pilocytic astrocytomas based on alterations other than mutations of the tumor suppressor gene p53. To prove these data, the authors screened a series of 42 astrocytic human brain tumors with a relatively high proportion (16.6%) of the pilocytic variant for the presence of p53 mutations, using the polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP) analysis, followed by DNA sequencing. Mutations were found in one of seven (14.3%) pilocytic astrocytomas, in one of 18 (5.6%) low grade astrocytomas, and in one of four (25%) anaplastic astrocytomas, but in none of 13 glioblastomas. Sites of missense mutations were in exon 8 (codons 281 and 282), and exon 5 (codon 151). Silent mutation was found in exon 9 (codon 324), which was related to pilocytic astrocytoma. This is, to the authors' knowledge, the first report that shows a p53 mutation in pilocytic astrocytomas. However, the p53 mutation was only found in one of seven tumors of this entity and was a silent mutation, which does not lead to change of amino acids. Thus, the significance of this alteration for the development of this special tumor type seems to be low. Nevertheless, it may be a sign of genetic instability and is thus suggested to be of certain pathogenetic relevance. The p53 findings concerning the other tumors are in accordance with the view of p53 gene mutations to be early events in astrocytoma formation.

Effect of structure on biological tolerance. Comparison of triiodobenzene derivatives.

Synthesis of basic structures of contrast media is reviewed. Biological tolerance of ionic and nonionic derivatives of triiodobenzene are related to the degree of withdrawal of electrons from their aromatic systems.

p53 mutation and protein alteration in 50 gliomas. Retrospective study by DNA-sequencing techniques and immunohistochemistry.

Alterations of the p53 protein, which is a 53 kD phosphoprotein and gene product of the p53 gene, has been found to play a major role in the genesis of a variety of human malignancies including tumors of the central nervous system. We investigated 50 tumor specimens from primary central nervous system neoplasms. Tissue samples were screened for mutations by the single-strand conformation polymorphism method and detected mutations were sequenced. All tissue specimens were stained immunohistochemically for p53 protein, which when altered accumulates in the nucleus due to prolonged half-life. Mutations were found in six cases, including one pilocytic astrocytoma World Health Organization (WHO) grade I, two astrocytomas WHO grade II, two anaplastic astrocytomas WHO grade III, and one primitive neuroectodermal tumor (PNET). In terms of relative frequency mutations were found mostly in the group of anaplastic astrocytomas WHO grade III. Interestingly, no mutations were found in the group of investigated glioblastomas. P53 immunopositivity did not correlated with the mutations found, whereas the staining index was significantly higher in the cases with detected mutations than in those without. When p53 alterations is seen as an indicator for different pathogenic pathways in glioma formation, this study gives evidence for a difference between anaplastic astrocytoma and glioblastoma. However, since there was a great overlap in p53 immunopositivity and p53 mutation in tumors of different WHO grades and entities, it seems that p53 will not act as a marker molecule neither for tumor entities nor for tumor malignancy.

Synthesis and Structure of a New Macrocyclic Polyhydroxylated Gadolinium Chelate Used as a Contrast Agent for Magnetic Resonance Imaging.

Three approaches to the synthesis of a new ligand 1,4,7-tris(carboxymethyl)-10-(1-(hydroxymethyl)-2,3-dihydroxypropyl)-1,4,7,10-tetraazacyclododecane (6) are described. This ligand forms the both thermodynamically and kinetically very stable gadolinium chelate Gadobutrol (1), which is a neutral and highly hydrophilic compound that is used for magnetic resonance imaging in the clinic. According to the crystal structure the Gd(III) ion in 1 is nine coordinated. The ligand provides eight coordination sites whereas the ninth coordination partner surprisingly is a carboxylate oxygen of a neighboring centrosymmetrically-related complex molecule. Ligand 6 was also utilized to prepare the calcium complex 12 which is used as an additive in the pharmaceutical formulation of 1. For the calcium complex 12, two complex molecules adopting almost identical conformations are present in the crystal.

Some physicochemical properties of the gadolinium-DTPA complex, a contrast agent for MRI.

The physicochemical characteristics of a good contrast agent for NMR are met by the gadolinium-DTPA complex. Measurements of density, viscosity and osmotic pressure of solutions to be injected are reported. A short survey of the results of X-ray analysis of the di-sodium-Gd-DTPA is given.

Trihalogenated benzamides--a new class of artificial sweeteners.

2,4,6-tribromo-3-carboxyalkyl-benzamides and -carboxyalkoxy-benzamides are intensely sweet. With a recognition threshold value of about 1 mumol/l the 3-carboxyethyl compound is one of the sweetest compounds known. The intensity of the sweet taste depends upon the nature of the carboxyalkyl/carboxylalkoxy group. In respect to the acute toxicity to mice the compounds are similar to the sodium salt of saccharin and sodium cyclamate. The compounds are easily obtainable in good yield.

A new lipophilic gadolinium chelate as a tissue-specific contrast medium for MRI.

Gd-Ethoxybenzyl-DTPA (Gd-EOB-DTPA) is a highly water-soluble paramagnetic contrast agent. Due to its protein binding of about 10% and its lipophilic residue, Gd-EOB-DTPA exhibits both renal (30% of the dose) and hepatobiliary (70%) excretion in rats. Despite its lipophilic character, the compound displays a low toxicity (LD50 = 7.5 mmol/kg). T1-relaxivity of 5.3 liters mmol-1 s-1 in water, 8.7 liters mmol-1 s-1 in plasma, and 16.9 liters mmol-1 s-1 in rat liver together with the hepatocellular uptake explain the liver-specific contrast enhancement of Gd-EOB-DTPA. The diagnostic dose is considerably lower than the amount of Magnevist used in abdominal imaging. The preclinical studies suggest its clinical role as being a hepatobiliary contrast agent for MRI.