A Comprehensive Patient-Derived Xenograft Collection Representing the Heterogeneity of Melanoma

Therapy of advanced melanoma is changing dramatically. Following mutational and biological subclassification of this heterogeneous cancer, several targeted and immune therapies were approved and increased survival significantly. To facilitate further advancements through pre-clinical in vivo modeling, we have established 459 patient-derived xenografts (PDX) and live tissue samples from 384 patients representing the full spectrum of clinical, therapeutic, mutational, and biological heterogeneity of melanoma. PDX have been characterized using targeted sequencing and protein arrays and are clinically annotated. This exhaustive live tissue resource includes PDX from 57 samples resistant to targeted therapy, 61 samples from responders and non-responders to immune checkpoint blockade, and 31 samples from brain metastasis. Uveal, mucosal, and acral subtypes are represented as well. We show examples of pre-clinical trials that highlight how the PDX collection can be used to develop and optimize precision therapies, biomarkers of response, and the targeting of rare genetic subgroups

Insulin-like growth factor-I receptor inhibition by specific tyrosine kinase inhibitor NVP-AEW541 in endometroid and serous papillary endometrial cancer cell lines

Endometrial cancer is the most widespread gynecological cancer in Western countries and constitutes a major public health issue. The role of the insulin-like growth factor (IGF) system in endometrial biology as well as in endometrial cancer has been well established. The IGF-I receptor (IGF-IR) emerged in recent years as a promising therapeutic target in a number of cancers. NVP-AEW541 (Novartis Pharma) is a pyrrolo(2,3-d)pyrimidine derivative with specific IGF-IR tyrosine kinase inhibitory activity. NVP-AEW541 has been shown to specifically abrogate IGF-I-mediated IGF-IR autophosphorylation and to reduce activation of the IGF-IR downstream signaling pathways. The aim of the present study was to investigate the potential anti-proliferative activities of NVP-AEW541 in Type I (endometrioid) and Type II (uterine serous papillary endometrial carcinoma, USPC) endometrial cancer cell lines. Results obtained showed that NVP-AEW541 abolished the IGF-I stimulated IGF-IR phosphorylation in all of the cell lines investigated (ECC-1, Ishikawa, USPC-1, USPC-2), whereas it abolished AKT and ERK phosphorylation in ECC-1 and USPC-1 cells. Furthermore, the inhibitor prevented from IGF-I from exerting its antiapoptotic effect in ECC-1, USPC-1 and USPC-2 cells. In addition, proliferation assays showed that NVP-AEW541 caused a significant decrease in proliferation rate in all of the cell lines. NVP-AEW541 had no major effect on the insulin receptor. In summary, our results suggest that specific IGF-IR inhibition by NVP-AEW541 is a promising therapeutic tool in endometrial cancer

Inhibition of IGF1R activity enhances response to trastuzumab in HER-2 positive breast cancer cells

BACKGROUND: Although trastuzumab has improved the prognosis for HER-2 positive breast cancer, not all HER-2 positive breast tumours respond to trastuzumab treatment and those that initially respond frequently develop resistance. Insulin-like growth factor-1 receptor (IGF1R) signalling has been previously implicated in trastuzumab resistance. We tested IGF1R inhibition to determine if dual targeting of HER-2 and IGF1R improves response in cell line models of acquired trastuzumab resistance. METHODS: HER-2, IGF1R, phospho-HER-2, and phospho-IGF1R levels were measured by ELISA in parental and trastuzumab-resistant SKBR3 and BT474 cells. IGF1R signalling was targeted in these cells using siRNA and the tyrosine kinase inhibitor NVP-AEW541. RESULTS: IGF1R levels were significantly increased in the trastuzumab resistant model, SKBR3/Tr, compared to the parental SKBR3 cell line. In both the SKBR3/Tr and BT474/Tr cell lines, inhibition of IGF1R expression with siRNA or inhibition of tyrosine kinase activity by NVP-AEW541 significantly increased response to trastuzumab. The dual targeting approach also improved response in the parental SKBR3 cells but not in the BT474 parental cells. CONCLUSIONS: Our results confirm that IGF1R inhibition improves response to trastuzumab in resistant HER-2 positive breast cancer cells, and also suggest that dual targeting of IGF1R and HER-2 may improve response in some trastuzumab-sensitive cells

Discovery of dihydroisoquinolinone derivatives as novel inhibitors of the p53-MDM2 interaction with a distinct binding mode

Blocking the interaction between the p53 tumor suppressor and its regulatory protein MDM2 is a promising therapeutic concept under current investigation in oncology drug research. We report here the discovery of the first representatives of a new class of small molecule inhibitors of this protein-protein interaction: the dihydroisoquinolinones. Starting from an initial hit identified by virtual screening, a derivatization program has resulted in compound 11, a low nanomolar inhibitor of the p53-MDM2 interaction showing significant cellular activity. Initially based on a binding mode hypothesis, this effort was then guided by a X-ray co-crystal structure of MDM2 in complex with one of the synthesized analogues. The X-ray structure revealed an unprecedented binding mode for p53-MDM2 inhibitors

The insulin-like growth factor-I receptor kinase inhibitor, NVP-ADW742, suppresses survival and resistance to chemotherapy in acute myeloid leukemia cells

The overexpression of Insulin-like growth factor-I receptor (IGF-IR) is strongly suggested to exert a role in malignant transformation and survival promotion in solid tumors such as breast cancer and prostate cancer, while fewer studies investigated the pathophysiological role of Insulin-like growth factor-I (IGF-I) signaling in acute myeloid leukemia (AML). In this study, we firstly found that the IGF-IR expression level positively correlated with the blast counts in de novo AML patients. Moreover, we observered that inhibitor of IGF-IR by NVP-ADW742, a novel kinase inhibitor of IGF-IR, could induce apoptosis in both HL-60 cell line and primary AML blasts, However, no significant alteration in the cell cycle status was observed in HL-60 cells. Further studies showed that NVP-ADW742 induced Akt dephosphorylation and p38 phosphorylation, decreased the expression of anti-apoptotic protein bcl-2 in HL-60 cells. Besides, we also demostrated that NVP-ADW742 could synergize with ara-C to induce apoptosis in subsets primary drug-resistant AML blasts. Therefore, IGF-IR targeting might be of therapeutic benefit for some AML patients

Pharmacokinetic–pharmacodynamic guided optimisation of dose and schedule of CGM097, an HDM2 inhibitor, in preclinical and clinical studies

Background: CGM097 inhibits the p53-HDM2 interaction leading to downstream p53 activation. Preclinical in vivo studies support clinical exploration while providing preliminary evidence for dosing regimens. This first-in-human phase I study aimed at assessing the safety, MTD, PK/PD and preliminary antitumor activity of CGM097 in advanced solid tumour patients (NCT01760525). Methods: Fifty-one patients received oral treatment with CGM097 10-400 mg 3qw (n = 31) or 300-700 mg 3qw 2 weeks on/1 week off (n = 20). Choice of dose regimen was guided by PD biomarkers, and quantitative models describing the effect of CGM097 on circulating platelet and PD kinetics. Results: No dose-limiting toxicities were reported in any regimens. The most common treatment-related grade 3/4 AEs were haematologic events. PK/PD models well described the time course of platelet and serum GDF-15 changes, providing a tool to predict response to CGM097 for dose-limiting thrombocytopenia and GDF-15 biomarker. The disease control rate was 39%, including one partial response and 19 patients in stable disease. Twenty patients had a cumulative treatment duration of >16 weeks, with eight patients on treatment for >32 weeks. The MTD was not determined. Conclusions: Despite delayed-onset thrombocytopenia frequently observed, the tolerability of CGM097 appears manageable. This study provided insights on dosing optimisation for next-generation HDM2 inhibitors. Translational relevance: Haematologic toxicity with delayed thrombocytopenia is a well-known on-target effect of HDM2 inhibitors. Here we have developed a PK/PD guided approach to optimise the dose and schedule of CGM097, a novel HDM2 inhibitor, using exposure, platelets and GDF-15, a known p53 downstream target to predict patients at higher risk to develop thrombocytopenia. While CGM097 had shown limited activity, with disease control rate of 39% and only one patient in partial response, the preliminary data from the first-in-human escalation study together with the PK/PD modeling provide important insights on how to optimize dosing of next generation HDM2 inhibitors to mitigate hematologic toxicity

Combined ALK and MDM2 inhibition increases antitumor activity and overcomes resistance in human ALK mutant neuroblastoma cell lines and xenograft models

The efficacy of ALK inhibitors in patients with ALK-mutant neuroblastoma is limited, highlighting the need to improve their effectiveness in these patients. To this end, we sought to develop a combination strategy to enhance the antitumor activity of ALK inhibitor monotherapy in human neuroblastoma cell lines and xenograft models expressing activated ALK. Herein, we report that combined inhibition of ALK and MDM2 induced a complementary set of anti-proliferative and pro-apoptotic proteins. Consequently, this combination treatment synergistically inhibited proliferation of TP53 wild-type neuroblastoma cells harboring ALK amplification or mutations in vitro, and resulted in complete and durable responses in neuroblastoma xenografts derived from these cells. We further demonstrate that concurrent inhibition of MDM2 and ALK was able to overcome ceritinib resistance conferred by MYCN upregulation in vitro and in vivo. Together, combined inhibition of ALK and MDM2 may provide an effective treatment for TP53 wild-type neuroblastoma with ALK aberrations

Discovery of a novel class of highly potent inhibitors of the p53–MDM2 interaction by structure-based design starting from a conformational argument

The p53–MDM2 interaction is an anticancer drug target under investigation in the clinic. Our compound NVP-CGM097 is one of the small molecule inhibitors of this protein–protein interaction currently evaluated in cancer patients. As part of our effort to identify new classes of p53–MDM2 inhibitors that could lead to additional clinical candidates, we report here the design of highly potent inhibitors having a pyrazolopyrrolidinone core structure. The conception of these new inhibitors originated in a consideration on the MDM2 bound conformation of the dihydroisoquinolinone class of inhibitors to which NVP-CGM097 belongs. This work forms the foundation of the discovery of HDM201, a second generation p53–MDM2 inhibitor that recently entered phase I clinical trial

Optimisation of a 5-[3-phenyl-(2-cyclic-ether)-methyl-ether]-4-aminopyrrolopyrimidine series of IGF-1R inhibitors

Taking the pyrrolopyrimidine derived IGF-1R inhibitor NVP-AEW541 as the starting point, the benzyl ether back-pocket binding moiety was replaced with a series of 2-cyclic ether methyl ethers leading to the identification of novel achiral [2.2.1]-bicyclic ether methyl ether containing analogues with improved IGF-1R activities and kinase selectivities. Further exploration of the series, including a fluorine scan of the 5-phenyl substituent, and optimisation of the sugar-pocket binding moiety identified compound 41 containing (S)-2-tetrahydrofuran methyl ether 6-fluorophenyl ether back-pocket, and cis-N-Ac-Pip sugar-pocket binding groups. Compound 41 showed improved selectivity and pharmacokinetics compared to NVP-AEW541, and produced comparable in vivo efficacy to linsitinib in inhibiting the growth of an IGF-1R dependent tumor xenograft model in the mouse

First-in-Human, Phase I Dose-Escalation Study of CGM097, a HDM2 Inhibitor in Adult Patients With p53 Wild-type Advanced Solid Malignancies

Background: CGM097 inhibits p53-HDM2 interaction, thus activates p53 downstream effector pathways inducing cell cycle arrest and/or apoptosis. This phase I study aimed at assessing the safety, MTD, PK/PD, and preliminary antitumor activity of CGM097 in advanced solid tumors patients (NCT01760525). Methods: Fifty-one patients received oral treatment with CGM097 10-400mg 3qw (n=31) and on an alternative regimen of 3qw 2 weeks on/1 week off (300-700mg; n=20). Choice of dose regimen was guided by PD biomarkers, PK/PD relationship, and modeling of drug-induced changes in platelet kinetics. Results: No dose-limiting toxicities were reported in any regimens. Grade 3/4 AEs suspected to be drug-related were reported in 21 patients. Main reason for discontinuation was disease progression (n=41). CGM097 plasma concentrations increased in a dose proportional manner. Disease control rate was 39%, including one partial response (300mg, 3qw) and 19 patients in stable disease (10-700mg 3qw). Forty patients had a cumulative treatment duration of >16 weeks, with 8 patients on treatment for >32 weeks. The MTD was not determined. Conclusions: Despite delayed-onset thrombocytopenia frequently observed, the tolerability of CGM097 appears manageable. Although CGM097 is not being further developed, this study led to important modeling-derived learnings to optimize dose scheduling of next generation HDM2 inhibitors