[ANSA analysis. II. Aminonaphthalenesulfonamides--detecting groups for polysubstrate analysis of proteases]. / ANSA-analiz. II. Aminonaftalinsul'fonamidy--detektiruemye gruppy dlia polisubstratnogo analiza proteaz.

The properties and synthetic methods of aminonaphthalenesulfonamides (ANSA) used as detectable groups of protease substrates are described. A list of chemical and physical properties of seventeen 5.1-ANSA with simple substituents is presented. A comparison of condition for the introduction and removal of acyl protecting groups (acetyl, trifluoroacetyl, phthaloyl, carbobenzoxy) used in ANSA synthesis is given. Examples of applicability of nitronaphthalenesulfonamides as intermediate compounds are given. The possibility of ANSA alkylation at both N(C) and N(S) is demonstrated. Substituted ANSA--sulfonylaziridenes--are used for the production of water-soluble derivatives containing the alcoxy group in the sulfonamide fragment. Criteria for the selection of detectable groups for polysubstrate analysis are discussed. Eighteen typical procedures for ANSA synthesis according to the schemes discussed are presented.

[ANSA analysis. III. Synthesis of aminonaphthalinesulfonamide chromogenic substrates for protease analysis]. / ANSA-analiz. III. Sintez aminonaftalinsul'fonamidnykh khromogreenykh substratov dlia analiza proteaz.

A review of synthetic methods of peptide substrates containing aminonaphthalenesulphonamide (ANSA) as the detected leaving group is presented. Variations of aminoacylic and peptide ANSA derivatives using ANSA as the C-protect group at all stages of the peptide synthesis, condensations of the ANSA with the N-protected peptide fragment obtained preliminary, the application of aminoacyl-ANSA as syntones are discussed. The synthesis scheme used while determining optimal ANSA substrates that involves reactions of aminoacyl derivatives of aminonaphthalenesulfonylchlorides with amines is shown. The application of di-tert-butylpyrocarbonate, DCC, chlorodimethylformiminium chloride, alkylchloroformate as condensing agents is described. The protection of amino groups was carried out by using Boc- and Cbz- groups.

[ANSA analysis. VI. Activation, inhibition and interaction of proteases in enzyme mixtures]. / ANSA-analiz. VI. Aktivatsiia, ingibirovanie i vzaimodeistviia proteaz v smesiakh fermentov.

A new accurate method (ANSA-analysis) is used for studying the interactions of proteases with their inhibitors or other proteases. The method is based on the cleavage by proteases of mixtures of competing chromogenic substrates containing aminonaphthalenesulfonamide (ANSA) detectable groups. Each substrate contained a specifically substituted ANSA group which showed its specific retention time during chromatographic separation. For the analysis of blood coagulation, mixtures of blood-clotting factor substrates were used. Hydrolysis of the substrate mixture catalyzed by blood samples gave characteristic chromatograms (ANSA spectra) for each sample. The activation time before injection of the blood sample into the substrate mixture and the pool of clotting factors and inhibitors both had influence upon the ANSA spectrum. The ANSA spectra of mixtures of trypsin and/or chymotrypsin with snake venoms are described as A x (the ANSA spectrum of a protease) + B x (the ANSA spectrum of a venom) + C x (the ANSA spectrum of catalytically active interaction products). They are additive (A = B = 1, C = 0), if no proteolysis, inhibition or activation takes place. ANSA spectra analysis shows deviations from additivity for some mixtures of Viperidae, (including E. carinatus), Naja naja, Agkistrodon contortrix and A. halys venoms. Explanations for the inability to detect inhibitors in venoms having a high protease activity by previously used methods are given.