Provided herein are hemoglobin-based therapeutic agents useful for treating cancer, pharmaceutical composition including the same, and methods of use and preparation thereof.

The invention provides methods, compositions, and medical kits comprising a nitrite-reductase promoter, such as an allosteric modulator of hemoglobin, for use in treating medical disorders and preservation of blood products. In one aspect, the invention provides methods, compositions, and medical kits comprising an inorganic nitrite salt and a nitrite-reductase promoter, such as an allosteric modulator of hemoglobin, for use in treating medical disorders, such as cancer, cardiovascular disorders, ischemic conditions, hemolytic conditions, and bacterial infections. Exemplary inorganic nitrite salts include sodium nitrite and arginine nitrite. Exemplary allosteric modulators of hemoglobin described herein include alkyl-substituted and acyl-substituted di-nitroheterocycles.

The use of hemoglobin as a targeting carrier for drugs is disclosed. Such hemoglobin-drug complexes have utility in the treatment of cancer, in particular cancers of the liver and colon. Said drug is preferably a nucleoside analog, in particular floxuridine and is covalently attached to the hemoglobin in a molar drug ratio of between 1 and 20.

Described herein are semisynthetic biopolymers comprising: a plurality of polyalkylene glycol chains, a protein, and an antioxidant; wherein the polyalkylene glycol chains are conjugated to the protein through a substituted succinimide linker. In some embodiments, the compounds described herein are conjugated proteins referred to as “semisynthetic supra perfusion agents”, “semisynthetic hybrid biopolymers”, “semisynthetic supra plasma expanders”, or the like. In some embodiments, these compounds mimic the same physiological consequences of high viscosity supra plasma expanders without being highly viscous—i.e. having a viscosity greater than blood.

The beta 2 subunit of mouse hemoglobin (HBB2) has been identified as soluble factor from mouse lungs that exhibits cytostatic/cytotoxic activity against neuroblastoma lung micrometastases. The beta subunit of human hemoglobin (HBB) has been found to have similar activity. Methods of using these proteins and fragments thereof in the treatment of cancer and inhibition of metastasis are provided, along with methods of screening a subject for micrometastases by detecting HBB in a biological sample.

The present invention provides a pharmaceutical composition containing recombinant hemoglobin protein or tetramer or dimer or subunit for tissue oxygenation and treating cancer. The recombinant hemoglobin protein or tetramer or dimer or subunit-based therapeutic agent is also effective for treating cancer. The recombinant hemoglobin or tetramer or dimer or its subunit moiety can target cancer cells and the therapeutic moiety (i.e. active agent/therapeutic drug) can kill the cancer cells efficiently. The recombinant hemoglobin or tetramer or dimer or its subunit-based therapeutic agent used in the present invention can be used in the treatment of various cancers such as pancreatic cancer, leukemia, head and neck cancer, colorectal cancer, lung cancer, breast cancer, liver cancer, nasopharyngeal cancer, esophageal cancer, prostate cancer, stomach cancer and brain cancer. The composition can be used alone or in combination with other therapeutic agent(s) such as chemotherapeutic agent, radiotherapeutic agent, anti-cancer protein drug to give a synergistic effect on cancer treatment, inhibiting metastasis and/or reducing recurrence.

The present invention relates to an isolated cellular targeted delivery system comprising a CD45.sup.+ leukocyte cell comprising within said cell a complex of one or more iron binding proteins and an active ingredient as well as methods for producing such isolated cellular targeted delivery system and uses of such system for therapy, in particular for therapy of cancer.

The present invention relates to an isolated cellular targeted delivery system comprising a CD45+ leukocyte cell comprising within said cell a complex of one or more iron binding proteins and an active pharmaceutically active substance and/or label as well as methods for producing such isolated cellular targeted delivery system and uses of such system for prophylaxis, therapy, diagnosis or theragnosis, in particular for prophylactic or therapeutic vaccination, therapy of cancer, particularly metastatic cancer or inflammatory diseases.

Provided herein are apohemoglobin-haptoglobin complexes as well as apohemoglobin-haptoglobin complexes comprising an active agent coordinated thereto. Methods of using these compositions are also described.

The disclosure concerns a class of hemoglobin based oxygen carriers (HBOCs) comprising a first hemoglobin protein cross-linked by a chemical reaction that is followed by a strain-promoted alkyne-azide cycloaddition (SPAAC) reaction or by a strain-promoted alkyne-nitrone cycloaddition (SPANC) in the absence of added copper salts to a second modified cross-linked hemoglobin protein. The resulting construct is an HBOC that is capable of binding oxygen and releasing same in a useful manner upon addition in an appropriate solution to the circulatory system of a patient. The disclosure also concerns a method of production of the HBOC where a first and second hemoglobin protein are produced by covalently linking hemoglobin to an angle strained cycloalkyne moiety. A compound comprising at least 2 azide or nitrone moieties is then added for reacting under conditions conducive to SPAAC or SPANC in reaction in absence of copper ions with the first and second hemoglobin protein for causing covalent linkage of the first and second hemoglobin protein to the compound comprising said at least 2 azide or nitrone moieties, resulting in said HBOC. The HBOC can be used for transfusion, perfusion or for increasing oxygen transport.