Hepatocellular carcinoma (HCC) is the 5th most common cancer type and one of the most aggressive tumors with poor recovery rates. Sorafenib (SOR) is a potent multiple kinase inhibitor drug assigned by FDA as the drug of choice for resistant HCC. Midkine (MK) is a heparin-binding cytokine known to be overexpressed in several malignant tumors with mitogenic, angiogenic, anti-apoptotic and chemo-resistant functions. The aim of the current study was to design a nano-sized lipid-based co-delivery system encapsulating both SOR and a small interfering RNA (siRNA) against MK gene to create a synergistic formulation for maximal efficacy. Various lipids were screened and the highest efficiency was obtained with YSK05, a novel pH-sensitive lipid synthesized in our laboratory. Cell viability studies confirmed synergism between SOR and MK-siRNA with efficient gene silencing as demonstrated by quantitative real time polymerase chain reaction (qRT-PCR) studies. Optimization of different formulation variables created an efficient system with minimum carrier’s toxicity. To improve system’s specificity towards HCC, the system was decorated with a novel targeting ligand exclusive for HCC, namely SP94 peptide, and further optimization has been carried out to balance efficiency and specificity. The system was tested on HCC cell line, HepG2, versus normal hepatocytes, FL83B. Cellular uptake studies by flow cytometry indicated a significant difference between HCC and normal hepatocytes in the uptake of the optimized system proving system’s selectivity for HCC. Cell viability data confirmed the high efficiency and specificity of the optimized system in HCC cells with a minimum effect on normal hepatocytes. The current data are promising and will be extrapolated to the in vivo level in the future.

Hepatocellular carcinoma (HCC) is the 5th most common cancer type and one of the most aggressive tumors with poor recovery rates. Sorafenib (SOR) is a potent multiple kinase inhibitor drug assigned by FDA as the drug of choice for resistant HCC. Midkine (MK) is a heparin-binding cytokine known to be overexpressed in several malignant tumors with mitogenic, angiogenic, anti-apoptotic and chemo-resistant functions. The aim of the current study was to design a nano-sized lipid-based co-delivery system encapsulating both SOR and a small interfering RNA (siRNA) against MK gene to create a synergistic formulation for maximal efficacy. Various lipids were screened and the highest efficiency was obtained with YSK05, a novel pH-sensitive lipid synthesized in our laboratory. Cell viability studies confirmed synergism between SOR and MK-siRNA with efficient gene silencing as demonstrated by quantitative real time polymerase chain reaction (qRT-PCR) studies. Optimization of different formulation variables created an efficient system with minimum carrier’s toxicity. To improve system’s specificity towards HCC, the system was decorated with a novel targeting ligand exclusive for HCC, namely SP94 peptide, and further optimization has been carried out to balance efficiency and specificity. The system was tested on HCC cell line, HepG2, versus normal hepatocytes, FL83B. Cellular uptake studies by flow cytometry indicated a significant difference between HCC and normal hepatocytes in the uptake of the optimized system proving system’s selectivity for HCC. Cell viability data confirmed the high efficiency and specificity of the optimized system in HCC cells with a minimum effect on normal hepatocytes. The current data are promising and will be extrapolated to the in vivo level in the future.

Hepatocellular carcinoma (HCC) is the worldwide's fifth cause of death. In the present study, we designed a novel lipid-based nanoplatform for highly-specific and efficient co-delivery of the cytotoxic drug, sorafenib (SOR), and small interfering RNA against Midkine gene (MK-siRNA) to HCC cells. SOR is designated by the FDA as the drug of choice for resistant HCC. Midkine (MK) is a growth cytokine highly-expressed in HCC with multiple malignant functions. The system was modified with a specific peptide ligand for selective uptake into HCC cells. We showed the first evidence that the combination of SOR and MK-siRNA had a synergistic cytotoxic effect on HCC cells. Our laboratory’s novel pH-sensitive cationic lipid, YSK05, showed the highest efficiency among the investigated lipids. Moreover, optimization of formulation variables improved the system’s outcome and resulted in a highly-specific internalization in both human and mouse HCC cells, (HepG2 and Hepa 1-6, respectively) compared to a significantly-lower internalization in other cancerous cells from non-HCC origin (HeLa), and no internalization in normal hepatocytes (FL83B). Cell viability studies confirmed the same highly-selective cytotoxicity pattern. The nanoplatform developed in this study has high potential as a promising therapy for HCC with minimum effect on normal cells.

Hepatocellular carcinoma (HCC) is the worldwide's fifth cause of death. In the present study, we designed a novel lipid-based nanoplatform for highly-specific and efficient co-delivery of the cytotoxic drug, sorafenib (SOR), and small interfering RNA against Midkine gene (MK-siRNA) to HCC cells. SOR is designated by the FDA as the drug of choice for resistant HCC. Midkine (MK) is a growth cytokine highly-expressed in HCC with multiple malignant functions. The system was modified with a specific peptide ligand for selective uptake into HCC cells. We showed the first evidence that the combination of SOR and MK-siRNA had a synergistic cytotoxic effect on HCC cells. Our laboratory’s novel pH-sensitive cationic lipid, YSK05, showed the highest efficiency among the investigated lipids. Moreover, optimization of formulation variables improved the system’s outcome and resulted in a highly-specific internalization in both human and mouse HCC cells, (HepG2 and Hepa 1-6, respectively) compared to a significantly-lower internalization in other cancerous cells from non-HCC origin (HeLa), and no internalization in normal hepatocytes (FL83B). Cell viability studies confirmed the same highly-selective cytotoxicity pattern. The nanoplatform developed in this study has high potential as a promising therapy for HCC with minimum effect on normal cells.

Hepatocellular carcinoma (HCC) is the fifth most common malignancy type around the world. The current study aimed at the design of a novel smart liposome for efficient and highly-specific co-delivery of the cytotoxic drug, Sorafenib (SOR), and a small interfering RNA against Midkine gene (MK-siRNA) to HCC cells. SOR is a multiple kinase inhibitor approved as the drug of choice for resistant HCC, but it is limited by its non-selective cytotoxicity and liability for the emergence of chemoresistance. Midkine is a growth cytokine overexpressed in HCC cells with several malignant functions including resistance to apoptosis. YSK05, a novel smart pH-sensitive cationic lipid synthesized in our laboratory, was selected as the main lipid for the formulation of the co-delivery liposome and showed superior efficiency over other investigated lipids in terms of both chemotherapy and gene therapy. Optimization of various formulation variables improved the efficiency and biosafety of the liposome. Furthermore, the introduction of a novel targeting peptide, SP94, imparted high selectivity to hepatic cancer cells with a minimal effect on normal hepatocytes. Selectivity was confirmed in terms of cellular uptake, cytotoxicity and gene knockdown activities. Moreover, we showed the first evidence for synergism between SOR and MK-siRNA via the increase of HCC cell sensitivity to SOR by MK-siRNA. In addition, our liposome showed a pH-sensitive controlled release pattern of SOR which secures prolonged action and minimizes off-target cytotoxicity. It can be concluded that the SP94-modified smart liposome is a promising co-delivery system for the targeted combinational treatment of HCC.

Hepatocellular carcinoma (HCC) is the fifth most common malignancy type around the world. The current study aimed at the design of a novel smart liposome for efficient and highly-specific co-delivery of the cytotoxic drug, Sorafenib (SOR), and a small interfering RNA against Midkine gene (MK-siRNA) to HCC cells. SOR is a multiple kinase inhibitor approved as the drug of choice for resistant HCC, but it is limited by its non-selective cytotoxicity and liability for the emergence of chemoresistance. Midkine is a growth cytokine overexpressed in HCC cells with several malignant functions including resistance to apoptosis. YSK05, a novel smart pH-sensitive cationic lipid synthesized in our laboratory, was selected as the main lipid for the formulation of the co-delivery liposome and showed superior efficiency over other investigated lipids in terms of both chemotherapy and gene therapy. Optimization of various formulation variables improved the efficiency and biosafety of the liposome. Furthermore, the introduction of a novel targeting peptide, SP94, imparted high selectivity to hepatic cancer cells with a minimal effect on normal hepatocytes. Selectivity was confirmed in terms of cellular uptake, cytotoxicity and gene knockdown activities. Moreover, we showed the first evidence for synergism between SOR and MK-siRNA via the increase of HCC cell sensitivity to SOR by MK-siRNA. In addition, our liposome showed a pH-sensitive controlled release pattern of SOR which secures prolonged action and minimizes off-target cytotoxicity. It can be concluded that the SP94-modified smart liposome is a promising co-delivery system for the targeted combinational treatment of HCC.

Novel quinoline derivatives carrying nitrones and oxime as nitric oxide donors were prepared and characterized using different spectroscopic techniques. Nitrones can release nitric oxide in larger amounts compared to corresponding oximes. Antiproliferative screening results showed that the 2-benzylthioquinoline nitrones 6e and 6f and the 2-methylthio analogues 6g and 6h exhibited promising antiproliferative activity especially against leukemia and colon cancer cell lines. Compounds 6c, 6e, and 6f exhibited higher potency as anticancer agents compared to doxorubicin, with IC50 ranging from 0.45 to 0.91 μM. Aremarkable overexpression of caspase-3 protein levels was observed in cells treated with the tested compounds. Compound 6e exhibited more pre-G1 apoptosis and cell cycle arrest at the G2/M phase than in other phases. These results revealed that the tested compounds can cause programmed cell death through overexpression of caspase 3, which may be attributed to the release of nitric oxide.

A series of novel compounds carrying 1,2,4-triazole scaffold were prepared and evaluated for their antiproliferative activities against NCI 60 cell line. Compounds 10 (a, c), 11 (a-d), and 14 (a-e) were selected for evaluation at single concentration of 10 µM towards panel of sixty cancer cell lines. Some of nitric oxide (NO) donating triazole/oxime hybrids 11a-d showed antiproliferative activity better than their corresponding ketones. On the other hand, the thiazolo [3,2-b][1,2,4]-triazoles 14a-e showed remarkable antiproliferative activities against the same cell lines. Compound 14d was selected for five dose testing against the full panel of 60 human tumor cell lines. Compound 14d showed high selectivity against renal subpanel with selectivity ratio of 6.99 at GI50 level. Compounds 11a-d, 10a-d and 14a-e were tested against four cell lines using MTT assay then compounds of the least IC50 were evaluated against three known anticancer targets including EGFR, BRAF and Tubulin. The results revealed that compound 14d showed promising EGFR inhibitory activity of cancer cell proliferation and were also observed to be moderate BRAF and tubulin inhibitors. Moreover, cell cycle analysis and apoptosis assay were finished for compounds 14d and 14f. Finally molecular modeling studies were performed to explore the binding mode of the most active compounds to the target enzymes.

A series of novel compounds carrying 1,2,4-triazole scaffold were prepared and evaluated for their antiproliferative activities against NCI 60 cell line. Compounds 10 (a, c), 11 (a-d), and 14 (a-e) were selected for evaluation at single concentration of 10 µM towards panel of sixty cancer cell lines. Some of nitric oxide (NO) donating triazole/oxime hybrids 11a-d showed antiproliferative activity better than their corresponding ketones. On the other hand, the thiazolo [3,2-b][1,2,4]-triazoles 14a-e showed remarkable antiproliferative activities against the same cell lines. Compound 14d was selected for five dose testing against the full panel of 60 human tumor cell lines. Compound 14d showed high selectivity against renal subpanel with selectivity ratio of 6.99 at GI50 level. Compounds 11a-d, 10a-d and 14a-e were tested against four cell lines using MTT assay then compounds of the least IC50 were evaluated against three known anticancer targets including EGFR, BRAF and Tubulin. The results revealed that compound 14d showed promising EGFR inhibitory activity of cancer cell proliferation and were also observed to be moderate BRAF and tubulin inhibitors. Moreover, cell cycle analysis and apoptosis assay were finished for compounds 14d and 14f. Finally molecular modeling studies were performed to explore the binding mode of the most active compounds to the target enzymes.

A novel quinolinyl pyrrolone and quinolinyl pyridazinone derivatives has been synthesized and characterized using different spectroscopic and elemental analysis techniques. Most of the target compounds displayed promising antiproliferative activity; In general, the pyrrolone derivatives 4a-f exhibited higher antiproliferative activity than their corresponding pyridazinone. The pyrrolone 4f showed outstanding antiproliferative activity with moderate selectivity against CNS and renal cancer with selectivity ratio of 3.49 and 3.56, respectively. Compound 4e and 5d experienced tubulin polymerization inhibitory activity comparable to that of vincristine while 4c, 4e and 4d showed good BRAF kinase inhibition compared to Erlotinib. Docking of compound 4e into colchicine binding site and biological assay results revealed that these compounds act mainly through tubulin polymerization inhibitory mechanism and can exhibit pre G1 apoptosis and cell cycle arrest at G2/M phase.