In vivo-stable bis-iminobiotin for targeted radionuclide delivery with the mutant streptavidin.

Targeted delivery of radionuclides to tumors is significant in theranostics applications for precision medicine. Pre-targeting, in which a tumor-targeting vehicle and a radionuclide-loaded effector small molecule are administered separately, holds promise since it can reduce unnecessary internal radiation exposure of healthy cells and can minimize radiation decay. The success of the pre-targeting delivery requires an in vivo-stable tumor-targeting vehicle selectively binding to tumor antigens and an in vivo-stable small molecule effector selectively binding to the vehicle accumulated on the tumor. We previously reported a drug delivery system composed of a low-immunogenic streptavidin with weakened affinity to endogenous biotin and a bis-iminobiotin with high affinity to the engineered streptavidin. It was, however, unknown whether the bis-iminobiotin is stable in vivo when administered alone for the pre-targeting applications. Here we report a new in vivo-stable bis-iminobiotin derivative. The keys to success were the identification of the degradation site of the original bis-iminobiotin treated with mouse plasma and the structural modification of the degradation site. We disclosed the successful pre-targeting delivery of astatine-211 (211At), α-particle emitter, to the CEACAM5-positive tumor in xenograft mouse models.

Abnormal Michael Reaction: Condition-Dependent Product Distributions in the Michael Reaction of Monoalkyl-Substituted Malonates.

The abnormal Michael reaction reported by Thorpe and Michael in 1900, which involves the transfer of an activating group from a nucleophilic species to an α-carbon of the Michael acceptor in reaction with monosubstituted malonates, was revisited using prototypical substrates for both intramolecular and intermolecular reactions. In both reactions, condition-dependent product distributions were observed. Thus, under the conditions using NaHMDS or NaH in THF or Et2O, no formation of the abnormal Michael product was observed, and the major product was an apparent retrograde Michael reaction in the abnormal Michael product resulting from an elimination of a malonate anion with the migrated acyl group as a part of it. The use of a base capable of generating a proton source such as NaOEt resulted in formation of the abnormal Michael product in the intermolecular reaction, although the retrograde Michael product was still a major product. On the other hand, in the intramolecular reaction, the abnormal product was not detected under any conditions used. A plausible reaction mechanism in which the lower kinetic acidity of the malonate proton compared to the thermodynamic acidity plays a significant role has been proposed.

Isolation and Reactions of Imidoyl Fluorides Generated from Oxime Using the Diethylaminosulfur Trifluoride/Tetrahydrofuran (DAST-THF) System.

Fluorination of oximes with the relatively mild diethylaminosulfur trifluoride/tetrahydrofuran (DAST-THF) system affords imidoyl fluorides. These compounds were isolated, and their structures were confirmed by X-ray single-crystal structure analysis. Reaction of imidoyl fluorides with various nucleophiles efficiently afforded amides, amidines, thioamides, and amine derivatives in high yields. Furthermore, one-pot reaction of in situ generated imidoyl fluorides from oximes was also applicable to efficient synthesis of these products. The oxime stereochemistry and acid-labile protecting group remained intact in this system.