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NPS-1034, a novel MET inhibitor, inhibits the activated MET receptor and its constitutively active mutants

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Summary

The MET proto-oncogene product, which is the receptor for hepatocyte growth factor (HGF), has been implicated in tumorigenesis and metastatic progression. Point mutations in MET lead to the aberrant activation of the receptor in many types of human malignancies, and the deregulated activity of MET has been correlated with tumor growth, invasion, and metastasis. MET has therefore attracted considerable attention as a potential target in anticancer therapy. Here, we report that a novel MET kinase inhibitor, NPS-1034, inhibits various constitutively active mutant forms of MET as well as HGF-activated wild-type MET. NPS-1034 inhibited the proliferation of cells expressing activated MET and promoted the regression of tumors formed from such cells in a mouse xenograft model through anti-angiogenic and pro-apoptotic actions. NPS-1034 also inhibited HGF-stimulated activation of MET signaling in the presence or absence of serum. Furthermore, when tested on 27 different MET variants, NPS-1034 inhibited 15 of the 17 MET variants that exhibited autophosphorylation with nanomolar potency; only the F1218I and M1149T variants were not inhibited by NPS-1034. Notably, NPS-1034 inhibited three MET variants that are resistant to the MET inhibitors SU11274, NVP-BVU972, and PHA665752. Together, these results suggest that NPS-1034 can be used as a potent therapeutic agent for human malignancies bearing MET point mutations or expressing activated MET.

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Abbreviations

HGF:

Hepatocyte growth factor

RTK:

Receptor tyrosine kinase

PI3K:

Phosphatidylinositide-3-kinase

MAPK:

Mitogen-activated protein kinase

SMA:

Smooth muscle actin

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Disclosure of potential conflicts of interests

Yoon SJ is an employee of NeoPharm Co., Ltd., which has ownership interest, including patents, in NPS-1034.

Funding

This study was supported by grants from the Korea Health 21 R&D Project, Ministry of Health and Welfare and Family Affairs, Republic of Korea (A062254 & HI10C2014).

Author information

Authors and Affiliations

  1. Institute for Innovative Cancer Research, University of Ulsan College of Medicine, Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul, 138-736, Seoul, Republic of Korea

    Jae-Sik Shin, Seung-Woo Hong, Jai-Hee Moon, Jin-Sun Kim, Kyung-Ah Jung, Seung-Mi Kim, Dae-Hee Lee, Chang-Gyu Lee, Eun-Kyoung Choi, Joo-Young Lee, Kyu-pyo Kim, Yong Sang Hong, Jae-Lyun Lee, Bongcheol Kim, Eun Kyung Choi, Jung Shin Lee, Dong-Hoon Jin & Tae Won Kim

  2. Department of Oncology, University of Ulsan College of Medicine, Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul, 138-736, Seoul, Republic of Korea

    Jae-Sik Shin, Seung-Woo Hong, Jai-Hee Moon, Jin-Sun Kim, Kyung-Ah Jung, Seung-Mi Kim, Dae-Hee Lee, Chang-Gyu Lee, Eun-Kyoung Choi, Joo-Young Lee, Kyu-pyo Kim, Yong Sang Hong, Jae-Lyun Lee, Jung Shin Lee, Dong-Hoon Jin & Tae Won Kim

  3. Department of Radiation Oncology, University of Ulsan College of Medicine, Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul, 138-736, Seoul, Republic of Korea

    Eun Kyung Choi

  4. Asan Institute for Life Science, University of Ulsan College of Medicine, Asan Medical Center, 88, Olympic-ro 43-gil, Songpa-gu, Seoul, 138-736, Seoul, Republic of Korea

    InKi Kim

  5. Department of New Drug Discovery, NeoPharm Co., Ltd., 928 Tamnip-Dong, Yuseong-Gu, Daejeon, 305-510, Republic of Korea

    Seon-Joo Yoon

Authors
  1. Jae-Sik Shin
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  2. Seung-Woo Hong
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  3. Jai-Hee Moon
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  4. Jin-Sun Kim
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  5. Kyung-Ah Jung
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  6. Seung-Mi Kim
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  8. InKi Kim
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  9. Seon-Joo Yoon
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  14. Yong Sang Hong
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  18. Jung Shin Lee
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  19. Dong-Hoon Jin
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  20. Tae Won Kim
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Corresponding authors

Correspondence to Dong-Hoon Jin or Tae Won Kim.

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Author contribution

Jae-Sik Shin, Seung-Woo Hong, and Jai-Hee Moon performed the majority of the experiments. Jin-Sun Kim, Kyung-Ah Jung, Seung-Mi Kim, Dae-Hee Lee, Chang-Gyu Lee, Eun-Kyoung Choi, and Joo-Young Lee helped with the experiments. InKi Kim, Seon-Joo Yoon, Kyu-pyo Kim, Yong-Sang Hong, Jae-Lyun Lee, Bongcheol Kim, Eun Kyung Choi, Jung Shin Lee, Dong-Hoon Jin, and Tae Won Kim analyzed the data. Dong-Hoon Jin and Tae Won Kim planned the experiments and wrote the manuscript.

Jae-Sik Shin, Seung-Woo Hong and Jai-Hee Moon contributed equally to this work.

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Supplementary Table S1

Analysis of MET autophosphorylation after ectopic expression of the 29 MET point mutants at Tyr-1234/1235 and Tyr-1349. (GIF 15 kb)

High Resolution Image (EPS 464 kb)

Supplementary Fig. S1

NPS-1034 inhibits HGF-stimulated signaling in prostate cancer cells. a PC3 and b DU145 cells were incubated with NPS-1034 or PHA665752 with or without fetal bovine serum for 2 h, followed by treatment with HGF (25 ng/ml) for 15 min. The effects of NPS-1034 on the signaling molecules downstream of MET were evaluated by Western blot analysis using antibodies against p-MET, MET, p-AKT, p-ERK, and γ-tubulin. (GIF 23 kb)

High Resolution Image (EPS 440 kb)

Supplementary Fig. S2

Analysis of body weights after NPS-1034 treatment. (GIF 6 kb)

High Resolution Image (EPS 370 kb)

Supplementary Fig. S3

Sequencing analysis of the various point mutants of MET were constructed from MET cDNA using site-directed mutagenesis. a The histogram displays the 29 MET point mutants using Sanger sequencing methods. b The 29 MET point mutants and wild-type MET cDNA expression was confirmed by RT-PCR. c AGS cells were transfected with the expression constructs of the 29 MET point mutants and the wild-type MET cDNA. The cell lysates were analyzed using antibodies against p-MET (Y1234/1235), p-MET (Y1349), MET, and γ-tubulin. (GIF 545 kb)

High Resolution Image (EPS 425 kb)

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Shin, JS., Hong, SW., Moon, JH. et al. NPS-1034, a novel MET inhibitor, inhibits the activated MET receptor and its constitutively active mutants. Invest New Drugs 32, 389–399 (2014). https://doi.org/10.1007/s10637-013-0039-4

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  • DOI: https://doi.org/10.1007/s10637-013-0039-4

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