Synthesis of photoactivable inhibitors of osteoclast vacuolar ATPase.
Amides of (2Z,4E)-5-[(5,6-dichloroindol-2-yl)]-2-methoxy-N-[3-[4-[3-(carboxymethoxy)phenyl)] piperazin-1-yl]propyl]-2,4-pentadienamide (1) and of 5-(5,6-dichloro-2-indolyl)-2-methoxy-2,4-pentadienoic acid (2) are strong inhibitors of the vacuolar ATPase located on the plasma membrane of osteoclasts. In order to understand which V-ATPase subunit is involved in the interaction with these novel inhibitors, analogues containing a photoactivable group and an iodine atom were designed. A series of alcohols or amines containing the photoactivable trifluoroaziridinophenyl or benzophenone moiety and an iodine atom were linked to the above acids via an ester or amide group. These compounds could be thereafter used as a radioactive photoprobe to label the protein. Whereas the compounds containing the photoactivable groups maintained good inhibitory activity, the introduction of the bulky iodine atom was generally detrimental, decreasing potency significantly. Better results were obtained by linking 3-(4-aminopiperidinomethyl)-3'-iodobenzophenone to 3-ethoxy-4-(2-(5,6-dichlorobenzimidazolyl))benzoic acid to give the corresponding amide 27, that inhibited vacuolar ATP-ase with a IC(50)=140 nM. The feasibility of introducing a radioactive 125I atom was ascertained by exchanging the iodine with a tributylstannyl group, that was again substituted by iodine.
Biasotti
Barbara
B
Dipartimento di Scienze Molecolari Agroalimentari, Università di Milano, Via Celoria 2, 20133 Milan, Italy.
Dallavalle
Sabrina
S
Merlini
Lucio
L
Farina
Carlo
C
Gagliardi
Stefania
S
Parini
Carlo
C
Belfiore
Pietro
P
D000577:Amides / Q000138:chemical synthesis*; D000818:Animals; D001562:Benzimidazoles / Q000737:chemistry; D019817:Benzoic Acid / Q000737:chemistry; D001577:Benzophenones / Q000737:chemistry; D001862:Bone Resorption / Q000201:enzymology; D045744:Cell Line, Tumor; D002645:Chickens; D004791:Enzyme Inhibitors / Q000138:chemical synthesis* / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D007211:Indoles / Q000138:chemical synthesis / Q000494:pharmacology; D007457:Iodine Radioisotopes; D007700:Kinetics; D015394:Molecular Structure; D010010:Osteoclasts / Q000187:drug effects / Q000201:enzymology; D019920:Photoaffinity Labels; D010880:Piperidines / Q000138:chemical synthesis / Q000494:pharmacology; D013329:Structure-Activity Relationship; D025262:Vacuolar Proton-Translocating ATPases / Q000037:antagonists & inhibitors*
C110962:5,6-dichlorobenzimidazole; D000577:Amides; D001562:Benzimidazoles; D001577:Benzophenones; D004791:Enzyme Inhibitors; D007211:Indoles; D007457:Iodine Radioisotopes; D019920:Photoaffinity Labels; D010880:Piperidines; C118025:SB 242784; C047723:benzophenone; D019817:Benzoic Acid; D025262:Vacuolar Proton-Translocating ATPases