Collagen-polysaccharide matrix for treatment of bone tumors

Drug – bio-affecting and body treating compositions – Preparations characterized by special physical form – Matrices

Reexamination Certificate

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C424S484000

Reexamination Certificate

active

06309670

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to crosslinked collagen-polysaccharide matrices for the treatment of bone tumors and methods of treatment using the matrices. The present invention provides a crosslinked collagen-polysaccharide matrix that is administered together with a differentiation factor for treatment of bone tumors.
BACKGROUND OF THE INVENTION
Bone tumors represents the major cause of morbidity and mortality associated with many types of cancer. Currently, the conventional procedure for treatment of bone tumor involves a combination of surgical resection, radiation and/or chemotherapy. Despite recent advances in the management of neoplastic diseases, the prognosis and quality of life for patients with aggressive bone tumors undergoing such multimodal therapies is still poor. Current experimental approaches for treatment of tumors are focused on the localized delivery of inhibitors of growth and neovascularization.
Collagens and glycosaminoglycans are two classes of biomaterials suited for use in bone regeneration. Collagen based matrices have been used in bone grafting. Type I collagen has good cell adhesive properties, in particular, for bone forming osteoblast cells.
Hyaluronic acid is a natural component of the extracellular matrix in many tissues including bone, and it is readily sterilized, is biodegradable and can be produced in a wide range of consistencies and formats. It is generally biocompatable and its resorption characteristics can be controlled by the manipulation of monomers to polymer forms, most commonly through the esterification of the carboxylic groups of the glucuronic acid residues.
The TGF-&bgr; superfamily consists of a large group of growth factors that exert profound influences on cellular morphology and the growth and differentiation of many cell types both in vitro and in vivo. The members of this superfamily are distinguished from other growth factors based on their unique ability to induce cell cycle arrest and the differentiation of mesenchymal cells and uncommitted cells during embryogenesis (Massague, J. S., Cheifetz, F. T., Andres, J. L. Ann. NY. Acad. Sci. 593:59-72, 1990). The mechanism of TGF-&bgr; induced cell cycle arrest is among the most intensively studied over the last decade. Members of this family include TGF-&bgr;1, &bgr;2 and &bgr;3, the Bone Morphogenetic Proteins (BMPs), and growth and differentiation factors (GDFs).
SUMMARY OF THE INVENTION
The present invention relates to methods of using a crosslinked collagen-polysaccharide matrice comprising a differentiation factor in the treatment of bone tumors. The present invention is based, in part, upon the unexpected finding that a collagen-polysaccharide matrix containing a differentiation factor inhibits growth of osteosarcoma cell lines more potently than the factor alone and the collagen-polysaccharide matrix alone. The present invention is also based upon the unexpected in vitro finding that a collagen-polysaccharide matrix containing TGF-&bgr; exhibits more potent anti-growth property than a combination of soluble collagen and TGF-&bgr;. The present invention is also based upon the finding that a collagen-polysaccharide matrix containing TGF-&bgr; elicits opposite responses in normal osteoblasts versus tumor cells.
Accordingly, the present invention provides a method for the treatment of bone tumors comprising the step of administering a matrix comprising (1) collagen covalently crosslinked to an exogenous polysaccharide, wherein said polysaccharide is crosslinked to said collagen through oxidized sugar rings on said polysaccharide which form covalent linkages to said collagen, and (2) a differentiation factor at a site of desired treatment As used in this discussion, an exogenous polysaccharide refers to a free polysaccharide.
In one embodiment, the matrix comprising the differentiation factor is implanted in a defect created after surgical resection of the tumor. In another embodiment, the matrix is injected in an osteolytic lesion.
The collagen used in a matrix of the present invention may be purified, native or modified collagen of any type. In one embodiment, the collagen is Type I collagen and in another embodiment, the collagen is Type II collagen.
The type of polysaccharides which can be used include hyaluronic acid, chondroitin sulfate, dermatan sulfate, keratan sulfate, heparan, heparan sulfate, dextran, dextran sulfate, alginate, and other long chain polysaccharides. In a preferred embodiment, the polysaccharide is hyaluronic acid.
The type of differentiation factor which can be used in the present invention include members of the TGF-&bgr; superfamily including, the Bone Morphogenic Proteins (BMPs); growth and differentiation factors (GDFs) and TGF-&bgr;1, B2 and B3. In a preferred embodiment, a crosslinked collagen-hyaluronic acid matrix comprising TGF-&bgr;1 is used in the treatment of bone tumors.
The ratios of the collagen to polysaccharide can be varied to change both the physical and biological properties of the matrix. A higher proportion of collagen will result in a more porous sponge-like matrix. A higher proportion of polysaccharide will result in a more gel-like matrix.
As used in this discussion, “treatment of bone tumor” refers to minimizing or eliminating the presence of tumor cells at the site of administration. Bone tumor includes osteosarcoma and neuroectodermal tumors. Administration encompasses injection of a gel-like matrix as well as implantation of a sponge like matrix.
As used in this discussion, “repair” is defined as growth of new tissue. The new tissue may or may not be phenotypically or genotypically identical to the original lost tissue. As used herein, “regeneration of tissue” means that the new tissue grown is identical to the lost tissue. Tissue repair can also be the result of replacing lost tissue with non-identical tissues. The basic cellular properties involved in repair include adhesion, proliferation, migration and differentiation.
By “conduction”, it is meant that the host tissue, e.g.,bone, grows by extension of existing tissue onto or into the crosslinked collagen-polysaccharide matrix. In conduction, repair cells move onto and into the matrix to synthesize and remodel new tissue identical to the surrounding host tissue. By induction, it is meant that the growth and differentiation of progenitor repair cells is stimulated. These progenitor cells go on to synthesize and remodel new tissue to be continuous with the surrounding host tissue.


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