TY - JOUR

T1 - Target enzyme mutations are the molecular basis for resistance towards pharmacological inhibition of nicotinamide phosphoribosyltransferase

A1 - Olesen,Uffe H

A1 - Petersen,Jakob G

A1 - Garten,Antje

A1 - Kiess,Wieland

A1 - Yoshino,Jun

A1 - Imai,Shin-Ichiro

A1 - Christensen,Mette K

A1 - Fristrup,Peter

A1 - Thougaard,Annemette V

A1 - Björkling,Fredrik

A1 - Jensen,Peter B

A1 - Nielsen,Søren J

A1 - Sehested,Maxwell

AU - Olesen,Uffe H

AU - Petersen,Jakob G

AU - Garten,Antje

AU - Kiess,Wieland

AU - Yoshino,Jun

AU - Imai,Shin-Ichiro

AU - Christensen,Mette K

AU - Fristrup,Peter

AU - Thougaard,Annemette V

AU - Björkling,Fredrik

AU - Jensen,Peter B

AU - Nielsen,Søren J

AU - Sehested,Maxwell

PB - BioMed Central Ltd.

PY - 2010

Y1 - 2010

N2 - BACKGROUND: Inhibitors of nicotinamide phosphoribosyltransferase (NAMPT) are promising cancer drugs currently in clinical trials in oncology, including APO866, CHS-828 and the CHS-828 prodrug EB1627/GMX1777, but cancer cell resistance to these drugs has not been studied in detail. METHODS: Here, we introduce an analogue of CHS-828 called TP201565 with increased potency in cellular assays. Further, we describe and characterize a panel of cell lines with acquired stable resistance towards several NAMPT inhibitors of 18 to 20,000 fold compared to their parental cell lines. RESULTS: We find that 4 out of 5 of the resistant sublines display mutations of NAMPT located in the vicinity of the active site or in the dimer interface of NAMPT. Furthermore, we show that these mutations are responsible for the resistance observed. All the resistant cell lines formed xenograft tumours in vivo. Also, we confirm CHS-828 and TP201565 as competitive inhibitors of NAMPT through docking studies and by NAMPT precipitation from cellular lysate by an analogue of TP201565 linked to sepharose. The NAMPT precipitation could be inhibited by addition of APO866. CONCLUSION: We found that CHS-828 and TP201565 are competitive inhibitors of NAMPT and that acquired resistance towards NAMPT inhibitors can be expected primarily to be caused by mutations in NAMPT.

AB - BACKGROUND: Inhibitors of nicotinamide phosphoribosyltransferase (NAMPT) are promising cancer drugs currently in clinical trials in oncology, including APO866, CHS-828 and the CHS-828 prodrug EB1627/GMX1777, but cancer cell resistance to these drugs has not been studied in detail. METHODS: Here, we introduce an analogue of CHS-828 called TP201565 with increased potency in cellular assays. Further, we describe and characterize a panel of cell lines with acquired stable resistance towards several NAMPT inhibitors of 18 to 20,000 fold compared to their parental cell lines. RESULTS: We find that 4 out of 5 of the resistant sublines display mutations of NAMPT located in the vicinity of the active site or in the dimer interface of NAMPT. Furthermore, we show that these mutations are responsible for the resistance observed. All the resistant cell lines formed xenograft tumours in vivo. Also, we confirm CHS-828 and TP201565 as competitive inhibitors of NAMPT through docking studies and by NAMPT precipitation from cellular lysate by an analogue of TP201565 linked to sepharose. The NAMPT precipitation could be inhibited by addition of APO866. CONCLUSION: We found that CHS-828 and TP201565 are competitive inhibitors of NAMPT and that acquired resistance towards NAMPT inhibitors can be expected primarily to be caused by mutations in NAMPT.

UR - http://www.biomedcentral.com.globalproxy.cvt.dk/1471-2407/10/677

U2 - 10.1186/1471-2407-10-677

DO - 10.1186/1471-2407-10-677

JO - B M C Cancer

JF - B M C Cancer

SN - 1471-2407

IS - 1

VL - 10

SP - 677

ER -