Hemoglobin-based oxygen carrier-containing pharmaceutical composition for cancer targeting treatment and prevention of cancer recurrence

Abstract

The present invention provides a pharmaceutical composition containing a hemoglobin-based oxygen carrier for treating cancer, preventing recurrence and metastasis of cancerous tumor. The composition can be used alone or in combination with at least one chemotherapeutic agent such as 5FU, Bortezomib, doxorubicin, cisplatin, or any combination thereof. The hemoglobin-based oxygen carrier in the composition is capable of targeting a surface receptor expressed on cancerous cells and facilitating the uptake of both hemoglobin-based oxygen carrier and the chemotherapeutic agent by the cancerous cells via a receptor-mediated mechanism. The hemoglobin-based oxygen carrier inhibits the expression of hypoxic response elements such as HIF1α, VEGF, ET1, VHL, etc. The pharmaceutical composition of the present invention is also useful for inducing the apoptosis or cell death of a type of self-renewing and tumor-initiating cells called cancer stem cells which are located in the hypoxic niche of a cancerous tumor.

1 . A hemoglobin-based oxygen carrier pharmaceutical composition configured to target a receptor expressed on the surface of cancer cells within a mass of cancerous tissue or tumors such that the hemoglobin-based oxygen carrier pharmaceutical composition triggers a receptor-mediated mechanism to facilitate the uptake and localization of the hemoglobin-based oxygen carrier into the cancer cells within the mass of the cancerous tissues or tumor for inducing apoptosis in cells of said cancerous tissues or tumors including self-renewing and tumor-initiating cells to prevent cancer recurrence, providing oxidative stress or shock to said cancerous tissues or tumors, and sensitizing the cancerous tissues or tumors to a chemotherapeutic agent or radiotherapy concurrently or subsequently administered. 
     
     
         2 . The pharmaceutical composition of  claim 1 , wherein said a chemotherapeutic agent is chemically conjugated with the hemoglobin-based oxygen carrier for synergistically targeting the cancerous tissues or tumors and the cancerous tumor cells including the self-renewing and tumor initiating cells. 
     
     
         3 . The pharmaceutical composition of  claim 1 , wherein said chemotherapeutic agent is 5 fluorouracil. 
     
     
         4 . The pharmaceutical composition of  claim 1 , wherein said hemoglobin-based oxygen carrier is a heat stable cross-linked tetrameric hemoglobin with an undetectable amount of dimer concentration and low concentration of met-hemoglobin. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein said receptor-mediated mechanism is Clathrin-mediated endocytosis. 
     
     
         6 . The pharmaceutical composition of  claim 1  is administered to a subject in need thereof prior to disruption of blood supply and/or during re-establishment of blood supply in a surgical removal of a tumor from said subject. 
     
     
         7 . The pharmaceutical composition of  claim 6  is administered in a range of approximately 0.2 g/kg-1.2 g/kg body weight of said subject. 
     
     
         8 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are hepatic. 
     
     
         9 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are hypoxic. 
     
     
         10 . The pharmaceutical composition of  claim 4 , wherein said cross-linked tetrameric hemoglobin has a molecular weight of 60-70 kDa and is heat treated with the addition of N-acetyl cysteine at a concentration of 0.05-0.4%. 
     
     
         11 . The pharmaceutical composition of  claim 1  is free of vasoconstricting impurities and protein impurities, non-pyrogenic, endotoxin-free, phospholipid-free, stroma-free and a met-hemoglobin level of less than 5%. 
     
     
         12 . The pharmaceutical composition of  claim 1 , wherein said hemoglobin-based oxygen carrier is modified having a longer half life to penetrate into the mass of said cancerous tissues or tumors and to provide oxidative stress or shock to said cancerous tissues or tumors such that apoptosis in cells of said cancerous tissues or tumors including said self-renewing and tumor-initiating cells is induced. 
     
     
         13 . The pharmaceutical composition of  claim 1 , wherein said self-renewing and tumor-initiating cells are cancer stem cells and/or progenitor cells. 
     
     
         14 . The pharmaceutical composition of  claim 12 , wherein said hemoglobin-based oxygen carrier is administered alone or in combination with at least one chemotherapeutic agent and/or radiotherapy, and wherein said hemoglobin-based oxygen carrier is administered as an adjunctive therapy. 
     
     
         15 . The pharmaceutical composition of  claim 33 , wherein said modified hemoglobin molecule is pegylated hemoglobin molecule. 
     
     
         16 . A method for treating cancerous tissues and preventing recurrence of cancerous tumors by using the composition of  claim 1 , wherein said composition is administered alone or in combination with at least one chemotherapeutic agent to a subject in need thereof. 
     
     
         17 . The method of  claim 16 , wherein said composition is administered to said subject during or after removal of cancerous tissues or tumors. 
     
     
         18 . The method of  claim 16 , wherein the cancerous tissues or tumors are hepatic, nasopharyngeal, brain, colon, lung, head and neck, mammary and leukemia. 
     
     
         19 . The method of  claim 16 , wherein the cancerous tissues or tumor are hypoxic. 
     
     
         20 . The method of  claim 16 , wherein said hemoglobin-based oxygen carrier is cross-linked tetrameric hemoglobin having a molecular weight of 60-70 kDa and is heat stable after heat treatment and addition of N-acetyl cysteine at a concentration of 0.05-0.4%. 
     
     
         21 . The method of  claim 20 , wherein said composition is free of vasoconstricting impurities and protein impurities, non-pyrogenic, endotoxin-free, phospholipid-free, stroma-free and a met-hemoglobin level of less than 5% after said heat treatment and reaction with the added N-acetyl cysteine. 
     
     
         22 . The method of  claim 16 , wherein said composition is administered by infusion in a range of approximately 0.2-1.2 g/kg body weight and at a rate of less than 10 ml/hour/kg body weight. 
     
     
         23 . The method of  claim 16 , wherein said at least one chemotherapeutic agent is selected from 5-fluorouracil, Bortezomib, doxorubicin, cisplatin, or any combination thereof. 
     
     
         24 . The method of  claim 16 , wherein said combination of the hemoglobin-based oxygen carrier and at least one chemotherapeutic agent are configured to synergistically target cells with expression of a receptor in said cancerous tissues or tumors, triggering a receptor-mediated mechanism, thereby sensitizing the cells in the cancerous tissues or tumors such that more hemoglobin-based oxygen carriers and chemotherapeutic agent are selectively taken up by the cells and localized in the cytoplasm of the cells while said cells become more sensitive to said chemotherapeutic agent. 
     
     
         25 . The method of  claim 16 , wherein said hemoglobin-based oxygen carrier is administered as an adjunctive therapy with said at least one chemotherapeutic agent for providing oxidative stress or shock to a mass of the cancerous tissues or tumors and for sensitizing said cancerous tissues or tumors to said chemotherapeutic agent such that apoptosis of cells in said cancerous tissues or tumors including self-renewing and tumor-initiating cells is induced. 
     
     
         26 . The method of  claim 25 , wherein said self-renewing and tumor-initiating cells are cancer stem cells and/or cancerous progenitor cells. 
     
     
         27 . (canceled) 
     
     
         28 . The method of  claim 24 , wherein said receptor-mediated mechanism is Clathrin-mediated endocytosis. 
     
     
         29 . The pharmaceutical composition of  claim 1 , wherein said chemotherapeutic agent is Bortezomib. 
     
     
         30 . The pharmaceutical composition of  claim 1 , wherein said chemotherapeutic agent is doxorubicin. 
     
     
         31 . The pharmaceutical composition of  claim 1 , wherein said chemotherapeutic agent is cisplatin. 
     
     
         32 . The pharmaceutical composition of  claim 1 , wherein said chemotherapeutic agent is more than one of 5-fluorouracil, Bortezomib, doxorubicin, and cisplatin. 
     
     
         33 . The pharmaceutical composition of  claim 1 , wherein said hemoglobin-based oxygen carrier is a polymerized hemoglobin with an undetectable amount of dimer concentration and low concentration of met-hemoglobin. 
     
     
         34 . The pharmaceutical composition of  claim 1 , wherein said hemoglobin-based oxygen carrier is a modified hemoglobin molecule with an undetectable amount of dimer concentration and low concentration of met-hemoglobin. 
     
     
         35 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are mammary. 
     
     
         36 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are brain cancerous tissues or tumors. 
     
     
         37 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are colon cancerous tissues or tumors. 
     
     
         38 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are lung cancerous tissues or tumors. 
     
     
         39 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are head and neck cancerous tissues or tumors. 
     
     
         40 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are nasopharyngeal. 
     
     
         41 . The pharmaceutical composition of  claim 1 , wherein said cancerous tissues or tumors are leukemia. 
     
     
         42 . The pharmaceutical composition of  claim 1 , wherein said hemoglobin-based oxygen carrier is configured to be administered as an adjunctive therapy with said chemotherapeutic agent for providing oxidative stress or shock to the mass of the cancerous tissues or tumors and for sensitizing said cancerous tissues or tumors to said chemotherapeutic agent such that apoptosis of cells in said cancerous tissues or tumors including self-renewing and tumor-initiating cells is induced.