US 9,809,595 B2
Flagellar and needle complex (injectosome) loop as anti bacterial drug target
Fadel Alexis Samatey, Kunigami-gun (JP); Vladimir A Meshcheryakov, Kunigami-gun (JP); Hideyuki Matsunami, Kunigami-gun (JP); and Akio Kitao, Tokyo (JP)
Assigned to OKINAWA INSTITUTE OF SCIENCE AND TECHNOLOGY SCHOOL CORPORATION, Okinawa (JP)
Appl. No. 14/770,708
Filed by OKINAWA INSTITUTE OF SCIENCE AND TECHNOLOGY SCHOOL CORPORATION, Kunigami-gun, Okinawa (JP)
PCT Filed Feb. 27, 2014, PCT No. PCT/JP2014/056082
§ 371(c)(1), (2) Date Aug. 26, 2015,
PCT Pub. No. WO2014/133197, PCT Pub. Date Sep. 4, 2014.
Claims priority of provisional application 61/770,046, filed on Feb. 27, 2013.
Prior Publication US 2016/0002239 A1, Jan. 7, 2016
Int. Cl. C07D 473/08 (2006.01); A61K 31/505 (2006.01); A61K 31/522 (2006.01); C12Q 1/02 (2006.01); C12Q 1/18 (2006.01); C07D 239/49 (2006.01)
CPC C07D 473/08 (2013.01) [A61K 31/505 (2013.01); A61K 31/522 (2013.01); C07D 239/49 (2013.01); C12Q 1/025 (2013.01); C12Q 1/18 (2013.01); G01N 2333/255 (2013.01); G01N 2500/04 (2013.01)] 4 Claims
 
1. A method for screening a compound that inhibits secretion of toxins into the host-cell cytoplasm by virulent bacteria using a needle type III secretion system, wherein the toxins are secreted by the needle type III secretion system and wherein the virulent bacteria is Salmonella typhimurium, the method comprising the steps of:
contacting a candidate compound with a C-terminal cytoplasmic domain of the membrane protein FlhB (SEQ ID NO: 1) from Salmonella typhimurium or an amino acid sequence variant that is at least 90% identical to SEQ ID NO: 1,
analyzing the interaction of the candidate compound with a loop region consisting of amino acids Glu, Asn, Lys, Met and Ser at positions 281 to 285 of FlhB (SEQ ID NO:1) or with the loop region of said variant that is at least 90% identical to SEQ ID NO: 1, wherein the loop region of said variant consists of amino acids Glu, Asn, Lys, Met and Ser, which correspond to positions 281 to 285 of FlhB (SEQ ID NO: 1), and
selecting a compound that reduces the flexibility of the loop region of FlhB or the amino acid sequence variant.