Bifunctional aryliodonium salts for highly efficient radioiodination and astatination of antibodies.

In this report we describe the development of an alternative approach to arylstannane chemistry for radiolabeling antibodies with radioiodine or astatine based on aryliodonium salts precursors. Bifunctional aryliodonium salts were designed and tested for the synthesis of 125I and 211At labeled prosthetic groups for bioconjugation. The nature of the electron rich aryl group was varied and its impact on the regioselectivity of radiohalogenation was evaluated. Unexpectedly, whereas the 2-thienyl group provided the best regioselectivity towards the radioiodination of the aryl moiety of interest (98:2), it was less selective for astatination (87:13); the anisyl group providing the best regioselectivity of astatination (94:6). Under optimized conditions, both radioiodination and astatination could be performed very efficiently in mild conditions (radiochemical yields>85%). The ionic nature of the precursors was exploited to develop an efficient purification approach: the HPLC step that is usually necessary in conventionnal approaches to optimize removal of organotin toxic precursors and side products was replaced by a filtration through a silica cartridge with a significantly reduced loss of radiolabeled product. The purified radioiodinated and astatinated prosthetic groups were then conjugated efficiently to an anti-CD138 monoclonal antibody (75-80% conjugation yield). By using this novel and simple radiohalogenation procedure, higher overall radiochemical yields of astatination were obtained in comparison with the use of an arylstannane precursor and procedures of the litterature for labeling the same antibody. Overall, due to their simplicity of use and high robustness, these new precursors should simplify the labeling of proteins of interest with iodine and astatine radioisotopes for imaging and therapeutic applications.

Investigation of the complexation of natZr(iv) and 89Zr(iv) by hydroxypyridinones for the development of chelators for PET imaging applications.

Three hydroxypyridinone (HOPO) positional isomers - 1,2-HOPO (L1H) and its water soluble analogue (L1'H), 3,2-HOPO (L2H) and 3,4-HOPO (L3H) have been investigated for the complexation of Zr(iv). Potentiometric and UV-Vis spectrometric studies show a higher thermodynamic stability for the formation of Zr(L1')4 in comparison with Zr(L2)4 and Zr(L3)4 as well as a higher kinetic inertness in competition studies with EDTA or Fe3+ at a radiotracer concentration with 89Zr. Besides the low pKa of L1H or L1'H (pKa = 5.01) in comparison with L2H and L3H (pKa = 8.83 and 9.55, respectively), the higher stability of Zr(L1')4 can be attributed in part to the presence of the amide group next to the chelating oxygen that induces intramolecular H-bond and amide/π interactions that were observed by X-ray crystallography and confirmed by quantum chemical calculations. The data presented here indicate that the 1,2-HOPO L1' exhibits the best characteristics for Zr(iv) complexation. However, 3,2-HOPO and 3,4-HOPO patterns, if appropriately tuned, for instance with the addition of an amide group as in the 1,2-HOPO ligand, may also become interesting alternatives for the design of Zr(iv) chelators with improved characteristics for applications in nuclear imaging with 89Zr.

Which radionuclide, carrier molecule and clinical indication for alpha-immunotherapy?

Beta-emitting radionuclides are not able to kill isolated tumor cells disseminated in the body, even if a high density of radiolabeled molecules can be targeted at the surface of these cells because the vast majority of emitted electrons deliver their energy outside the targeted cells. Alpha-particle emitting radionuclides may overcome this limitation. It is thus of primary importance to test and validate the radionuclide of choice, the most appropriate carrier molecule and the most promising clinical indication. Four α-particle emitting radionuclides have been or are clinically tested in phase I studies namely 213Bi, 225Ac, 212Pb and 211At. Clinical safety has been documented and encouraging efficacy has been shown for some of them (213Bi and 211At). 211At has been the most studied and could be the most promising radionuclide but 225Ac and 212Pb are also of potential great interest. Any carrier molecule that has been labeled with ß-emitting radionuclides could be labeled with alpha particle-emitting radionuclide using, for some of them, the same chelating agents. However, the physical half-life of the radionuclide should match the biological half-life of the radioconjugate or its catabolites. Finally everybody agrees, based on the quite short range of alpha particles, on the fact that the clinical indications for alpha-immunotherapy should be limited to the situation of disseminated minimal residual diseases made of small clusters of malignant cells or isolated tumor cells.

Application of response surface methodology to optimise the antioxidant activity of a saithe (Pollachius virens) hydrolysate.

The objective of this study was to produce, by an enzymatic hydrolysis process at a pilot scale, a saithe (Pollachius virens) hydrolysate with a high antioxidant activity. Design of experiment methodology, based on laboratory-scale experiments, was used to obtain a behavioral reduced model that allows one to determine the optimal operating conditions maximizing the antioxidant activity. Two specifications were studied: the degree of hydrolysis and the antioxidant activity. The effects of the following hydrolysis parameters (temperature, pH, enzyme concentration, and operating time) were studied and presented as response surfaces. From these results, a multifactorial optimization was performed and the Pareto optimal set of efficient solutions was evaluated. The optimal conditions were tested at laboratory scale and then validated by comparison with tests carried out on a pilot plant.

Evaluation of nitrogenous substrates such as peptones from fish:a new method based on Gompertz modeling of microbial growth.

Fish peptones from tuna, cod, salmon, and unspecified fish were compared with a casein one by using a new method based on Gompertz modeling of microbial growth. Cumulative results obtained from six species of bacteria, yeasts, and fungi showed that, in most cases, these fish peptones are very effective. Nevertheless, this study raised some questions about the standardization of fish raw material, the enzymatic hydrolysis of fish proteins, and the composition of the culture medium used for testing the peptones.

[The history of health in Quebec: relationships and specificities]. / L' histoire de la santé au Québec: filiations et spécificités.

The remarkable development of the history of health in Quebec during the last two decades occurred at the junction of two main historiographical movements: the international resurgence of the history of health, and the metamorphosis of the historian's gaze toward Quebec. The author examines specific features issued from this junction. He shows that many historians challenged the "retard du Québec" thesis. Studying, as they did, the relations between health service providers, they unveiled their respective interests and conflicts, which led to the invalidation of the idea of a monolithic backwards society.

Electrophoretic study of the caseinolytic activity of a pepsin from the dogfish Scyliorhinus canicula.

Electrophoretic patterns of casein and casein subfractions were studied following proteolysis by dogfish pepsin II or calf chymosin. Both enzymes hydrolyze the kappa casein subfraction with the production of kappa paracasein peptide. alpha S1 and beta subfractions hydrolysis is stronger with dogfish enzyme than with chymosin. It is concluded that, despite a broader specificity, the activity spectrum of dogfish enzyme is, in many respects, similar to that of calf chymosin.

Characterization of a chymosin-like pepsin from the dogfish Scyliorhinus canicula.

1. Pepsin II extracted from the gastric mucosa of Scyliorhinus canicula has been characterized and compared to calf chymosin. 2. The kcat and Km of the dogfish enzyme for the synthetic hexapeptide Leu-Ser-Phe(NO2)-Nle-Ala-Leu-OMe have been determined. The kcat/Km ratio is close to that of calf chymosin. Its milk-clotting efficiency is however 21-fold lower than that of calf chymosin. 3. The proteolytic activity against haemoglobin is optimal at pH 2.5. It clots the milk up to pH 6.8. 4. The dogfish pepsin II shows relatively better activity at low temperatures than calf chymosin.