Necuparanib, A Multitargeting Heparan Sulfate Mimetic, Targets Tumor and Stromal Compartments in Pancreatic Cancer

Abstract

Pancreatic cancer has an abysmal 5-year survival rate of 8%, making it a deadly disease with a need for novel therapies. Here we describe a multitargeting heparin-based mimetic, necuparanib, and its antitumor activity in both in vitro and in vivo models of pancreatic cancer. Necuparanib reduced tumor cell proliferation and invasion in a three-dimensional (3D) culture model; in vivo, it extended survival and reduced metastasis. Furthermore, proteomic analysis demonstrated that necuparanib altered the expression levels of multiple proteins involved in cancer-driving pathways including organ development, angiogenesis, proliferation, genomic stability, cellular energetics, and invasion and metastasis. One protein family known to be involved in invasion and metastasis and altered by necuparanib treatment was the matrix metalloprotease (MMP) family. Necuparanib reduced metalloproteinase 1 (MMP1) and increased tissue inhibitor of metalloproteinase 3 (TIMP3) protein levels and was found to increase RNA expression of TIMP3. MMP enzymatic activity was also found to be reduced in the 3D model. Finally, we confirmed necuparanib's in vivo activity by analyzing plasma samples of patients enrolled in a phase I/II study in patients with metastatic pancreatic cancer; treatment with necuparanib plus standard of care significantly increased TIMP3 plasma protein levels. Together, these results demonstrate necuparanib acts as a broad multitargeting therapeutic with in vitro and in vivo antiinvasive and antimetastatic activity.

Cancer Metastasis: The Role of the Extracellular Matrix and the Heparan Sulfate Proteoglycan Perlecan

“…In addition to roneparastat, HS mimetics muparfostat (PI-88) (233), neuparanib (N-402) (234), piixatimod (PG545) (251), have been, or are currently in clinical trials for use as a therapy targeting metastasis of melanoma or pancreatic cancer. More recent reports have detailed the use of these HS mimetics not only in the development of therapeutics, but the development of more representative models for testing anticancer/anti-metastatic therapeutics including patient-derived xenografts (237) and organoid models (238). Neuparanib has been shown to reduce tumor cell proliferation and invasion in an organoid model, and plasma levels of patients within a clinical trial cohort reported increased levels of tissue inhibitor of MMP-3 (238).…”

“…More recent reports have detailed the use of these HS mimetics not only in the development of therapeutics, but the development of more representative models for testing anticancer/anti-metastatic therapeutics including patient-derived xenografts (237) and organoid models (238). Neuparanib has been shown to reduce tumor cell proliferation and invasion in an organoid model, and plasma levels of patients within a clinical trial cohort reported increased levels of tissue inhibitor of MMP-3 (238). The attempt at mimicking the structure of HS has seen the development of glycopolymers with well-defined sulfation patterns and the ability to optimize disaccharide length for peak heparanase inhibition (252), which reduced metastasis of breast cancer in a rodent model.…”

The Challenge of Modulating Heparan Sulfate Turnover by Multitarget Heparin Derivatives

“…Analysis of plasma samples of patients with metastatic pancreatic cancer enrolled in a phase I/II study show that treatment with necuparanib in addition to the standard of care significantly increased TIMP3 plasma protein levels confirming the in vivo and in vitro studies on pancreatic cancer models [171]. PG545 has been selected over the PG500 series of heparan sulfate mimetics for the ability to inhibit both angiogenesis and heparanase activity and was selected as the lead clinical candidate for oncology [191].…”

“…Glycol-splitting of unsubstituted uronic acids obtained by periodate oxidation and borohydride reduction has been applied to a variety of heparin derivatives and increased the flexibility of the molecules for a better interaction and inhibition of heparanase or growth factors. M402 (Necuparanib) is a glycol-split LMWH with reduced anticoagulant activity [170] that was found to reduce tumor burden in vivo in KPFMC pancreatic cancer mice model at a dose of 40 mg/kg/day and reduce AsPC-1 pancreatic cancer cell line proliferation and invasion in vitro in a 3D-culture model [171]. Boothello et al synthesized a non-saccharide compound, G2.2, which is structurally homogeneous and easy to obtain by chemical synthesis, as a mimetic of an HS hexasaccharide active on cancer stem cells.…”

“…Interestingly, MacDonald et al observed that M402 also targeted the stromal compartment. On a co-culture of pancreatic tumor cells and stellate cells that mimics epithelial-stromal interactions, treatment of M402 significantly decreased the invasive behavior in a dose-dependent manner, whereas in vivo it extended survival and reduced metastasis by reducing the protein levels of matrix metalloprotease 1 and by increasing the tissue inhibitor of metalloproteinase 3 (TIMP3) [171]. Karoli et al demonstrated that carbohydrate-based HS-mimicking compounds, PI-88 and analogues, inhibit heparanase and compete with growth factors (FGF-1, FGF-2, and VEGF) for binding to HS, and therefore impact anti-angiogenic and anti-metastatic activity [48,181].…”