Synthèse de phosphoramidates de 2-désoxy-2-iodoglycosyles

Addition of iodazide to acetylated, benzylated, and methoxymethylated glycals yielded 2-deoxy-2-iodoglycosyl azides with 1,2-trans configuration. Stereoselectivity of the reaction favored the manno and talo configurations starting from d-glucal and d-galactal, respectively. With d-xylal derivatives, the stereoselectivity depended on the nature of the substituents. The Staudinger reaction of 2-deoxy-2-iodoglycosyl azides with trimethylphosphite led to the corresponding 2-deoxy-2-iodoglycosyl phosphoramidates in high yield.     

Synthèse des 2-acé-tamido-2-désoxy-6-O-α-et β-D-xylopyranosyl-α-D-glucopyranose

2-Acetamido-2- deoxy-6-O-, -xylopyranosyl-O-D-glucopyranose has been synthesized in crystalline form by condensation of 2,3,4-tri-O-acetyl-α-D-xylopyranosyl chloride (1) with benzyl 2-acetamido-3,4-di-O-acetyl-2-deoxy-β-D-glucopyranoside (2), followed by O-deacetylation and catalytic hydrogenation. Condensation of 2 with 2,3,4-tri-O-chlorosulfonyl-β-D-xylopyranosyl chloride, followed by dechlorosulfonylation and acetylation, gave benzyl 2-acetamido-3,4-di-O-acetyl-2-deoxy-6-O-(2,3,4-tri-O-acetyl-α-D-xylopyranosyl)β-D-glucopyranoside in crystalline form. O-Deacetylation, followed by catalytic hydrogenation, gave 2-acetamido-2-deoxy-6-O-α-D-xylopyranosyl-α-D-glucopyranose in crystalline form.     

Synthèse du méthyl-2-acétamido-2-désoxy-β-D-glucofuranoside et de dérivés

Methyl 2-acetamido-2-deoxy-5,6-O-isopropylidene-β-D-glucofuranoside was prepared in excellent yield from methyl 2-benzamido-2-deoxy-5,6-O-isopropylidene-β-D-glucofuranoside by alkaline hydrolysis, followed by selective N-acetylation. Treatment with 60% acetic acid at room temperature gave syrupy methyl 2-acetamido-2-deoxy-β-D-glucofuranoside, characterized by a crystalline tri-O-p-nitrobenzoyl derivative. The same treatment, at 100° gave methyl 2-acetamido-2-deoxy-β-D-glucopyranoside. In an alternative procedure, the selective N-acetylation was performed after acetic acid hydrolysis of methyl 2-amino-2-deoxy-5,6-O-isopropylidene-β-D-glucofuranoside. Several derivatives of methyl 2-acetamido-2-deoxy-β-D-glucofuranoside were prepared and compared with the corresponding pyranosides. The furanoside structure was clearly demonstrated by mass spectrometry and periodate oxidation.     

Synthèse des méthyl-2,4,6-tridésoxy-4-trifluoroacétamido-l-hexopyranosides. Accès à de nouvelles anthracyclines

The four diastereoisomeric methyl 2,4,6-trideoxy-4-trifluoroacetamido-l-hexopyranosides have been synthesized. Coupling of the corresponding 1-O-acetyl-3-O-benzoyl-l-lyxo and 1-O-acetyl-3-O-p-nitrobenzoyl-l-arabino derivatives with daunomycinone in the presence of p-toluensulfonic acid as catalyst afforded two new anthracyclines.     

Synthèse et étude R.M.N. de disaccharides et trisaccharides dans la série du l-rhamnose

Various di- and tri-saccharides containing l-rhamnose were synthesized by condensation of 2,3,4-tri-O-acetyl- or 2,3,4-tri-O-benzoyl-α-l-rhamnopyranosyl bromide with an unblocked glycopyranoside. The determination of the anomeric configuration of l-rhamnose saccharides by n.m.r. is difficult because structure has a greater effect on the spectra than does configuration. The α and β configurations and the position of the substitution may be assigned from the chemical shifts of H-5 and CH₃. In all the compounds having a β configuration, a shielding of the methyl group and a deshielding of the H-5 proton have been observed as compared to the compounds having an α configuration. The H-5 proton and the methyl group of peracetylated, (1→3)-linked α-l derivatives always resonate at higher fields than the corresponding protons of (1→6)-linked α-l derivatives.     

Synthèse de l'acide 5-acétamido-3,5-didésoxy-4-O-méthyl- d-glycéro-d-galacto-2-nonulosonique (acide 4-O-méthyl-N-acétylneuraminique). Partie II

3-Acetamido-3-deoxy-4,5:6,7-di-O-isopropylidene-d-glycero-d-galacto-heptose diethyl dithioacetal was transformed into 3-acetamido-3-deoxy-4,5:6,7-di-O-isopro-pylidene-2-O-methyl-aldehydro-d-glycero-d-galacto-heptose after O-methylation followed by desulfuration. A Wittig reaction with an excess of [ethoxy(ethoxycarbonyl)-methylene]triphenylphosphorane in the presence of benzoic acid gave a mixture of ethyl 5-acetamido-3.5-dideoxy-2-O-ethyl-6,7:8,9-di-O-isopropylidene-4-O-methyl-d-glycero-d-galacto-non-2-enonate (23 %) and the d-glycero-d-talo (22 %) isomer. An ethoxymercuration-demercuration reaction, followed by acid hydrolysis, converted the former into ethyl 4-O-methyl-N-acetylneuraminate and the latter into the C-4 stereoisomer. 4-O-Methyl-N-acetylneuraminic acid was then obtained in crystalline form, and its structure ascertained by mass spectrometry and ¹H- and ¹³C-nuclear magnetic resonance.     

Analyse de la fécondation par la parthénogénèse expérimentale

Sans résumé     

Imagerie de la néoangiogenèse en médecine nucléaire

The formation of new vessels, a process referred to as neoangiogenesis, is one of the key pathophysiological mechanisms in the development and progression of cancer. It contributes to tumour growth and dissemination of neoplastic cells and can determine response or resistance to anticancer therapies. It involves different signaling pathways including the vascular endothelial growth factor (VEGF) pathway and integrins, which are also preferred targets for the development of antiangiogenic therapies. Changes in the microvasculature induced by antiangiogenic treatments occur before morphological changes can be detected with conventional imaging approaches. The development of molecular tools enabling an assessment of these targets before initiating therapy, or early detection of response or recurrence during or following treatment is essential for the close monitoring of antiangiogenic treatments. These outstanding needs call for the development of specific probes enabling the characterization of the molecules and pathways involved. This review summarizes the major signaling pathway involved in promoting tumor neoangiogenes is, the different radiotracers recently developed in preclinical and clinical settings, as well as their potential use in humans in order to improve the management of patients treated with antiangiogenic treatments.     

Synthèse d'ARN et de protéines dans des disques de patte de drosophile cultivés in vitro

Résumé Des disques imaginaux de patte, prélevés dans des larves de fin de 3e stade de drosophile, ont été cultivés dans le milieu M de Mandaron, en l'absence d'hormone ou en présence soit d'- soit de -ecdysone. L'incorporation de précurseurs marqués (³H-uridine,³H-leucine ou³H-proline) a été étudiée en fonction du stade de développement des disques.En l'absence d'hormone de mue, l'incorporation d'uridine décroît dès que les disques ont été explantés; l'incorporation de leucine et de proline ne décroît que 6 à 12 heures après l'explantation.- et -ecdysone stimulent l'incorporation des trois précurseurs; toutefois celle-ci varie en fonction du développement morphologique du disque.Les maxima et les minima d'incorporation d'uridine précèdent dans le temps ceux de la leucine et de la proline.Les maxima d'incorporation peuvent être mis en rapport avec des évènements morphologiques marquants du développement: évagination, sécrétion des cuticules nymphale et maginale.Il n'y a pas de différences significatives d'incorporation d'uridine en présence d'-ecdysone ou de -ecdysone; en revanche les maxima d'incorporation de leucine et de proline sont plus élevés en présence d'-ecdysone que de -ecdysone.Ces résultats montrent que l'-ecdysone—et à un degré moindre la -ecdysone—peuvent induire les synthèses de macromolécules nécessaires au développement des appendices in vitro.

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RNA and protein synthesis inDrosophila leg discs cultured in vitro

Summary Imaginal leg discs from late third instarDrosophila larvae were cultured in Mandaron's medium without hormone or with -ecdysone or -ecdysone. Incorporation of labelled precursors (tritiated uridine, tritiated leucine or tritiated proline) was studied as a function of the stage of in vitro disc development.In the absence of moulting hormone, uridine incorporation decreased as soon as the discs were explanted; leucine and proline incorporation however began to decrease only after 6 to 12 h.- and -ecdysone stimulated the incorporation of all three precursors; however the rate of the incorporation varied as a function of the morphological disc development.The maxima and minima of uridine incorporation preceeded in time those of proline and leucine incorporation.The peaks of incorporation were coincident with salient morphological events of development: evagination, secretion of pupal and imaginal cuticles.There were no significant differences in uridine incorporation in the presence of -ecdysone or -ecdysone. Leucine and proline incorporation maxima however were significantly higher in the presence of -ecdysone than of -ecdysone.The results show that -ecdysone—and to a lesser extent also -ecdysone—can induce the macromolecular syntheses required for the development of the appendage in vitro.

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Ce travail a été réalisé avec l'aide du CNRS (Action thématique programmée «Différenciation cellulaire», contrat n^o A 6554324)

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Ce mémoire représente une partie de la thèse qui sera soutenue par l'auteur devant l'Université Scientifique et Médicale de Grenoble     

Facteurs impliqués dans le remodelage de la chromatine au cours de la spermiogenèse

Round spermatids are post-meiotic cells with a haploid genome contained in a nucleus, with a structure initially similar to that of the somatic cell nucleus. During spermatogenesis, the spermatid nucleus undergoes drastic remodelling during which it first elongates and then condenses into the very specific and tightly packaged structure of the sperm nucleus. During this remodelling dthe histones are replaced by transition proteins, which, in turn, are replaced by protamines, the specific nuclear proteins of the spermatozoa. Immediately prior to their replacement, the histones are hyperacetylated. The first part of our work was to precisely characterise the changes in histone acetylation during murine spermatogenesis. We have shown that the core histones H2A, H2B, H3 and H4 are hyperacetylated in the elongating spermatids. We have also shown that these changes in acetylation are associated with degradation of the enzymes responsible for histone deacetylation, histone deacetylases or HDACs, while histone acetyl transferases are still present in these cells. The histone acetylation pattern was also investigated during human spermatogenesis, revealing that histone hyperacetylation in the nucleus of elongating spermatids, which appears to be conserved during the course of evolution, also occurs during human spermatogenesis. Moreover, our data obtained from the testes of men with severely altered spermatogenesis, including SCO syndromes (Sertoli Cells Only Syndromes), show that a global hyperacetylation of the Sertoli cell nuclei is associated with an absence of meiotic and post-meiotic cells. This suggests that the global histone acetylation variations observed during spermatogenesis are part of a signalling pathway involving germ cell — Sertoli cell communication. Altogether, these data provide a basis for a better understanding of the mechanisms and identification of the factors involved in post-meiotic remodelling of chromatin.     

Étude de la cytodifférenciation du rein de l'Epinoche femelle après traitement par la méthyltestostérone

Résumé Au microscope électronique, l'action de la méthyltestostérone sur les cellules rénales de l'Epinoche femelle se traduit par une cytodifférenciation conduisant à la formation de cellules glandulaires muqueuses. Elle a lieu simultanément à deux niveaux distincts:Au niveau du tubule proximal, le premier signe visible de cytodifférenciation est une activation du nucléole, accompagnée par une augmentation de taille des cellules. Puis on assiste à un développement de l'ergastoplasme et de l'appareil de Golgi et à l'élaboration de deux types de sécrétions: d'abord des granules de 2000–2500 Å, ensuite des grains de mucigène typiques, qui subissent rapidement une transformation muqueuse.Une cytodifférenciation régressive intervient en même temps. Elle concerne la pinocytose apicale qui disparaît.Au niveau des tubules collecteurs, la cytodifférenciation se traduit par la formation d'un mucus hyalin d'origine golgienne. Elle s'accompagne également d'une disparition de la pinocytose.La méthyltestostérone est capable de provoquer, chez la femelle, une cytodifférenciation rénale identique à celle que l'on observe chez le mâle pendant la période de reproduction. La transformation muqueuse des cellules rénales est donc sous le contrôle de la seule testostérone, qui déclenche au niveau cellulaire un ensemble de processus conduisant à la formation de mucus.Au microscope électronique, on constate que l'élaboration de mucus prêt à l'excrétion est achevée au bout de trois jours dans les tubules proximaux alors que dans les tubules collecteurs elle ne demande que 24 heures.

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Action of methyl testosterone on the cytodifferentiation of the kidney of the female three-spined Stickleback

Summary At the microscopic level, the action of methyl testosterone on the cells of the kidney of the female three-spined Stickleback gives raise to a cytodifferentiation which leads to the formation of mucous glandular cells. This action is evident at two different levels:At the level of the proximal tubule, the first visible sign of cytodifferentiation is an activation of the nucleolus, accompanied by a growth of the cell size. Then a rapid development of the ergastoplasm and the Golgi apparatus takes place, which leads to the elaboration of two types of secretory particles: granules of 2000–2500 Å in diameter appear first, then typical mucigen granules become visible. These latter undergo a rapid mucous transformation.A regressive cytodifferentiation occurs at the same time. It concerns the apical pinocytosis which disappears in the cells undergoing the glandular differentiation.At the level of the collecting tubules, the cytodifferentiation is characterized by the elaboration of a clear mucus which originates in the Golgi apparatus and migrates to the apex of the cells. A disappearance of the pinocytosis is also noticeable.In the kidney of the female, methyl testosterone induces a cytodifferentiation which is identical to that occuring in the male during the breeding period. So the mucous transformation of renal cells is under the control of a single hormone: the testosterone, which is able to give raise to a succession of phenomena leading to the formation of a mucous secretion.With the electron microscope, it is possible to demonstrate that three days are necessary for the elaboration of the mucus ready to be discharged in the lumen of the renal proximal tubules. In the collecting tubules, the reaction occurs more quickly, after only two days of treatment.

L'auteur tient à remercier Monsieur le Professeur E. Follénius pour ses conseils et son aide précieuse au cours de la réalisation de ce travail.