Glycofullerenes as non-receptor tyrosine kinase inhibitors- towards better nanotherapeutics for pancreatic cancer treatment

Abstract

The water-soluble glycofullerenes GF1 and GF2 were synthesized using two-step modified Bingel-Hirsch methodology. Interestingly, we identified buckyballs as a novel class of non-receptor Src kinases inhibitors. The evaluated compounds were found to inhibit Fyn A and BTK proteins with IC 50 values in the low micromolar range, with the most active compound at 39 µM. Moreover, we have demonstrated that formation of protein corona on the surface of [60]fullerene derivatives is changing the landscape of their activity, tuning the selectivity of obtained carbon nanomaterials towards Fyn A and BTK kinases. The performed molecular biology studies revealed no cytotoxicity and no influence of engineered carbon nanomaterials on the cell cycle of PANC-1 and AsPC-1 cancer cell lines. Incubation with the tested compounds resulted in the cellular redox imbalance triggering the repair systems and influenced the changing of protein levels. Pancreatic cancer is an extremely aggressive type of cancer with a poor prognosis and a five-year survival rate of less than 7% 1. Moreover, only about 20% of patients are eligible for surgical resection, the only potentially curative treatment 2. Chemotherapeutic approaches, which are often based on gemcitabine, are still present in the clinic as the adjuvant therapy of choice for resected pancreatic cancer. The novel cytotoxic combination strategies and nanoformulations which include Folfirinox and nab-paclitaxel are being developed intensively, but these treatments have their own shortcomings and their therapeutic effects are disappointing 3. Nanomedical strategies which include metallofullerene-based (Gd@C 82 (OH) 22) inhibitors of matrix metalloproteinases (MMPs) 4 , photothermally active nanoparticles 5,6 , and several nanodelivery systems 7,8 are also being presented in attempts to improve survival in pancreatic cancer patients. At the same time, the pharmaceutical industry has focused significant attention around the use of selective tyrosine kinase inhibitors as anti-cancer agents 9. The EGFR kinase inhibitor Erlotinib has been approved by FDA for use in combination with gemcitabine for locally advanced or metastatic pancreatic cancer 10. Recently, some reports have suggested that inhibition of non-receptor intracellular Src kinases inhibits growth and metastasis of human pancreatic carcinoma in murine models, and is followed by synergetic effects in combination therapy with gemcitabine 11. In order to overcome the barriers to standard chemotherapeutics, we have been exploring the use of [60]fullerene derivatives as novel EPR-targeted nanotherapeutics in in vitro and in vivo models 12,13. Previously, we have reported the creation of photoactive and highly water-soluble glycofullerene GF2 (termed 'Sweet-C 60 ') that predominantly accumulates in the nucleus of pancreatic stellate cells (PSCs) and is inherently non-toxic even in high concentrations (above 1 mg/mL) 14. Results and Discussion Based on the above background, we have synthesized two glycofullerenes: GF1 and GF2 (see Fig. 1A) c...

The Landscape of the Anti-Kinase Activity of the IDH1 Inhibitors

“…The inhibition of the tyrosine kinases by the tested compounds was measured using the Kinase Selectivity TK-2 Profiling Systems (Promega) as was previously reported [68,69]. Briefly, the kinases (ABL1; BRK; BTK; CSK; Fyn A; Lck; Lyn B; Src) and substrate/co factor strips were prepared according to the manufacturer's instructions.…”

Novel Benzenesulfonate Scaffolds with a High Anticancer Activity and G2/M Cell Cycle Arrest

“…Assays using the Kinase Selectivity TK-2 profiling systems and ADP-Glo Kinase Assay (both from Promega) were performed to determine the inhibition of the non-receptor tyrosine kinases. The protocol was previously designed and described by our group in reports [ 15 , 80 ]. The experiments were performed at least four times.…”

Emerging Kinase Therapeutic Targets in Pancreatic Ductal Adenocarcinoma and Pancreatic Cancer Desmoplasia

“…For pancreatic cancer, highly specific pharmaceuticals targeting single kinases (rather than families or groups of kinases) remain underdeveloped. Currently, Bosutinib [ 80 ], 7f [ 251 ], Dapagliflozin [ 256 ], Masitinib [ 221 ], and glycofullerenes [ 257 ] have been studied as potentially useful kinase inhibitors in pancreatic cancer. Alternatively, agents such as proteolysis targeting chimeras (PROTACs) degrade—rather than inhibit—protein targets and may prove better mimics of genetic knockdown for instances in which nonenzymatic activity (e.g., activity as a ligand) underlie a kinase’s role in a particular disease [ 258 ].…”