A High Throughput Screen Identifies Nefopam as Targeting Cell Proliferation in β-Catenin Driven Neoplastic and Reactive Fibroproliferative Disorders

Abstract

Fibroproliferative disorders include neoplastic and reactive processes (e.g. desmoid tumor and hypertrophic scars). They are characterized by activation of β-catenin signaling, and effective pharmacologic approaches are lacking. Here we undertook a high throughput screen using human desmoid tumor cell cultures to identify agents that would inhibit cell viability in tumor cells but not normal fibroblasts. Agents were then tested in additional cell cultures for an effect on cell proliferation, apoptosis, and β-catenin protein level. Ultimately they were tested in Apc1638N mice, which develop desmoid tumors, as well as in wild type mice subjected to full thickness skin wounds. The screen identified Neofopam, as an agent that inhibited cell numbers to 42% of baseline in cell cultures from β-catenin driven fibroproliferative disorders. Nefopam decreased cell proliferation and β-catenin protein level to 50% of baseline in these same cell cultures. The half maximal effective concentration in-vitro was 0.5 uM and there was a plateau in the effect after 48 hours of treatment. Nefopam caused a 45% decline in tumor number, 33% decline in tumor volume, and a 40% decline in scar size when tested in mice. There was also a 50% decline in β-catenin level in-vivo. Nefopam targets β-catenin protein level in mesenchymal cells in-vitro and in-vivo, and may be an effective therapy for neoplastic and reactive processes driven by β-catenin mediated signaling.

Patient‐derived mouse xenografts from pediatric liver cancer predict tumor recurrence and advise clinical management

“…In agreement with SIOPEL's oncologists, we evaluated PDX response to treatments that could be proposed to children with recurrent disease, including irinotecan alone or in combination with temsirolimus or sirolimus, two mammalian target of rapamycin inhibitors, sorafenib, which is currently used in HCC in adults, crizotinib, given that its activity as a epithelial‐mesenchymal transition (EMT) inhibitor might counteract increased MET signaling previously described in HB, and paclitaxel, also extensively evaluated in pediatric solid tumors . In addition to these clinically well‐known compounds, we tested tumor response to Nefopam, a nonopioid analgesic drug, because it was shown to decrease CTNNB1 product β‐catenin expression in cancer cells . For all these targeted therapies, tumor response in the PDXs tested was highly insufficient.…”

“…(39) In addition to these clinically well-known compounds, we tested tumor response to Nefopam, a nonopioid analgesic drug, because it was shown to decrease CTNNB1 product b-catenin expression in cancer cells. (40) For all these targeted therapies, tumor response in the PDXs tested was highly insufficient. Irinotecan alone has been tested with promising results in a phase II trial of the SIOPEL group, (41) whereas irinotecan in combination with rapamycin (sirolimus) is currently administered in a phase I protocol for pediatric patients with refractory solid tumors (Clinical-Trials identifier: NCT01282697).…”

Pharmacologically targeting beta-catenin for NF1 associated deficiencies in fracture repair

“…Taking this limitation into consideration, both in vitro and in vivo models displayed enhanced cellular differentiation and matrix production however, previous work has indicated Nefopam treatment to lead to decreased cellular proliferation [28]. These observations work in concert with the idea of β-catenin playing a disparate role in bone healing.…”

“…Here we investigate a therapeutic intervention to improve bone repair in NF1, by targeting the Wnt/β-catenin signaling pathway using (RS)-5-methyl-1-phenyl-1,3,4,6-tetrahydro-2,5-benzoxazocine (Nefopam). Nefopam is an analgesic agent which also is a modulator of β-catenin signaling [28].…”

“…Nefopam was administered via an intraperitoneal injection (30mg/kg/day) on the day before, the day of, and the day after fracture surgery. Dosing analysis has previously been performed [28].…”