RP-R-02LM

Amgen StudyLung Micro-Metastases Diameter

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RP-R-02LM

Amgen StudyTotal Lung Metastases Count

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AMG 386

Inhibition of angiopoietin 1/2 and c-met impairs metastatic potential in a patient derived xenograft of renal carcinoma

Ashley Orillion2,4, Remi Adelaiye1,2, Li Shen2, Eric Ciamporcero2,3, Sheng Yu Ku1,2, Swathi Ramakrishnan1,2, May Elbanna5, Sreenivasulu Chintala2,5, and Roberto Pili1,2

1Department of Cancer Pathology and Prevention, Roswell Park Cancer Institute Division, University at Buffalo, Buffalo NY, USA; 2Genitourinary Program, Roswell Park Cancer Institute, Buffalo NY, USA; Department of Medicine, Simon Cancer Center, Indiana University, Indianapolis IN, USA: 3Department of Medicine and Experimental Oncology, University of Turin, Turin Italy; 4Department of Stress and Cellular Biology, Roswell

Park Cancer Institute Division, University at Buffalo NY, USA; 5Faculty of Pharmacokinetics and Pharmacodynamics, Roswell Park Cancer Institute, Buffalo NY, USA; 6Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo NY, USA.

Background: Clear cell renal cell carcinoma (ccRCC) is the most common form of kidney cancer and diagnoses have been on the rise since the 1990s. While patients with early stage ccRCC have more than a 50% survival rate over four years, those with advanced stage disease have a less than 10% chance of 5 year survival. Angiopoietins and c-MET pathway activation have been implicated in tumro angiogenesis and metastasis. In our current study, we utilized an angiopoietin 1/2 inhibitor (Amgen 386) and a novel c-MET inhibitor in murine models grafted with human derived xenografts (PDX) with a high metastatic penetrance. Our goal was to assess the ability of these compounds to prolong survival via metastatic inhibition. Methods: Seven week old SCID mice were implanted either subcutaneously or orthotopically with human ccRCC PDX RP-R-02LM. Approximately one month post implantation, mice were started on treatment with Angiopoietin 1/2 inhibitor (Amgen 386), c-met inhibitor (compound A) or a combination of the two. Amgen 386 was administered twice a week subcutaneously at 5.6mg/kg. Compound A was given daily via oral gavage at 30mg/kg. As a survival study, treatment was continued until the mice reached a point where they needed to be euthanized or they were found dead. Results: In our metastatic ccRCC PDX model, RP-R-02LM, Amgen386 single agent treatment and combination c-MET inhibitor and Amgen386 treatment significantly reduced metastases (p=0.0075 and p=0.0001 respectively). Further, an ongoing survival study has shown that at four and a half months post implantation both single agent and combination treatments prolong the survival of mice implanted with RP-R-02LM, a highly metastatic PDX. Conclusions: Our results show that the combination of Amgen386 and novel c-MET inhibitor, Compound A, significantly inhibits the spread of metastases and prolongs survival in our highly metastatic ccRCC PDX model. This indicates a potential use for combination therapy in treating patients with ccRCC.

ABSTRACT

INTRODUCTION

RESULTS

CONCLUSIONS

ACKNOWLEDGEMENTS

This study was supported by in part by the National Cancer Institute, National Institutes of Health (P30CA016056) (RP) and a research grant from Amgen (RP).

OBJECTIVES

• Our initial results pointed to the potential of both AMG386 as a single agent and in combination with a novel c-MET inhibitor, Compound A, as inhibitors of metastases in our patient derived xenograft lung metastatic model (RP-R-02LM).

• Our results show that, in a PDX model of metastatic ccRCC, Amgen 386 and Combination treatment of Amgen 386 and Compound A results in improved survival and reduced metastases.

• Overall, our current data suggests that there may be potential for treating some subtypes of ccRCC that have similar characteristics to our highly metastatic patient derived xenograft 2 – Lung Metastatic (RP-R-02LM) by using this combination approach.

• Assess the pre-clinical efficacy of Amgen 386, a novel angiopoietin 2 inhibitor, as an anti-angiogenic therapy for ccRCC patients with metastatic disease.

• Assess the pre-clinical role of Compound A, a novel c-MET inhibitor, in enhancing the anti-metastatic activity of Amgen 386.

• Assess the potential of AMG 386 and Compound A on extending survival in a metastatic renal carcinoma model.

Treatment of clear cell renal cell carcinoma continues to be present a clinical challenge due to tumor progression and driven by anti-VEGF resistance. Anti-VEGF resistance in ccRCC involves various growth factors including c-MET and those of the angiopoietin family.

Amgen 386 has been reported to neutralize angiopoietin 1 and 2, thus preventing the interaction of these molecules with their target receptors. This may result in an inhibition of angiogenesis and a potential inhibition tumor cell metastasis.

As a selective inhibitor of c-MET, Compound A, is expected to block c-MET activation. Thus Compound A should inhibit the proliferation, survival and migration of the ccRCC in our pre-clinical models.

We expected that this study may provide pre-clinical evidence of AMG 386 and Compound A anti-metastastic effects in ccRCC models and provide a rationale for a clinical trial in patients with metastatic kidney cancer.

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Proliferation MigrationCell Survival

Figure 1. AMG386 and Combination treatments yielded a significant reduction in the number and size of lung metastases. (A) Quantification of the macro- and micro-metastases showed a statistically significant reduction in metastases to the lungs when comparing Vehicle to both single agent treatment AMG386 (p=0.0337) and combination treatment (p=0.0004). Additionally, the combination treatment surpassed both single agent treatments in suppressing the metastases, with Amgen – Combination p=0.0036 and Compound A – Combination p=0.0012. (B) Lung tissues were paraffin embedded, sectioned and stained with hematoxylin and eosin stain to quantify the presence and diameter of micro-metastases. (C) In addition to a significant reduction in the number of metastases, the size of the lung metastases was significantly reduced when treated with single agent AMG 386 (p=0.0047) and in an additive manner when treated with Amgen 386 and Compound A together (p=0.0001). Results are expressed as the mean ± SE, n = mice per group. *p<0.05, **p<0.005.

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Figure 2. Amgen 386 single agent and Combination treatments prolong the life and reduce the metastatic burden of mice with metastatic renal cell carcinoma. (A) Survival curve of a subcutaneous RP-R-02LM study shows a significant shift in the survival of mice with metastatic ccRCC. (Vehicle-Amgen: *p-value = 0.0125; Vehicle-Compound A: p-value = not significant; Vehicle-Combination: p-value = 0.0250; Amgen-Compound A: p-value = 0.0381; Amgen-Combination: p-value = not significant; Compound A-Combination: p-value = 0.0318. (B) Lungs from mice in the survival study combination group have significantly fewer metastatic nodules at time of death than those in the vehicle

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Poster #2857