Recombinant poliovirus for the treatment of cancer

Drug – bio-affecting and body treating compositions – Whole live micro-organism – cell – or virus containing – Genetically modified micro-organism – cell – or virus

Reexamination Certificate

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C424S093600

Reexamination Certificate

active

06264940

ABSTRACT:

The present invention is directed to non-pathogenic, oncolytic, recombinant polioviruses for the treatment of various forms of malignant tumors. More particularly, the present invention is directed to the administration of the non-pathogenic, oncolytic, recombinant poliovirus to the tumor directly, intrathecally or intravenously to cause tumor necrosis. The method of the present invention is particularly useful for the treatment of malignant tumors in various organs, such as: breast, colon, bronchial passage, epithelial lining of the gastrointestinal, upper respiratory and genito-urinary tracts, liver, prostate and the brain. Astounding remissions in experimental animals have been demonstrated for the treatment of malignant glioblastoma multiforme, an almost universally fatal neoplasm of the central nervous system.
The invention was made with Government support under No. AI32100-07 and AI39485 awarded by the National Institutes of Health. The government has certain rights in the invention.
BACKGROUND OF THE INVENTION
Known Methods of Treatment
It has been known that malignant tumors result from the uncontrolled growth of cells in an organ. The tumors grow to an extent where normal organ function may be critically impaired by tumor invasion, replacement of functioning tissue, competition for essential resources and, frequently, metastatic spread to secondary sites. Malignant cancer is the second leading cause of mortality in the United States.
Up to the present, the methods for treating malignant tumors include surgical resection, radiation and/or chemotherapy. However, numerous malignancies respond poorly to all traditionally available treatment options and there are serious adverse side effects to the known and practiced methods. There has been much advancement to reduce the severity of the side effects while increasing the efficiency of commonly practiced treatment regimens. However, many problems remain, and there is a need to search for alternative modalities of treatment. The search is particularly urgent for primary malignant tumors of the central nervous system. Brain tumors, especially glioblastomas, remain one of the most difficult therapeutic challenges. Despite the application of surgery, radiotherapy and chemotherapy, alone and in combination, glioblastomas are almost always fatal, with a median survival rate of less than a year and 5-year survival rates of 5.5% or less. None of the available therapeutic modes has substantially changed the relentless progress of glioblastomas.
Systematic studies of patients who were diagnosed with malignant glioma and underwent surgery to wholly or partially remove the tumor with subsequent chemotherapy and/or radiation showed that the survival rate after 1 year remains very low, particularly for patients who are over 60 ears of age. Leibel, S. A., et al.,
Cancer,
35:1551-1557 (1975); Walker, M. D., et al.,
J. Neurosurg.,
49:333-343 (1978); Chang, C. H., et al.,
Cancer,
52:997-1007 (1983). Malignant gliomas have proven to be relatively resistant to radiation and chemotherapeutic regimens. Bloom, H. J. G.,
Int. J. Radiat. Oncol. Biol. Phys.,
8:1083-1087 (1982). Adding to the poor prognosis for malignant gliomas is the frequent tendency for local recurrence after surgical ablation and adjunct radiation/chemotherapy. Choucair, A. K., et al.,
J. Neurosurg.,
65:654-658 (1986).
Treatment of Cancer with Viruses
In recent years, there have been proposals to use viruses for the treatment of cancer: (1) as gene delivery vehicles, Miller, A. D.,
Nature,
357:455-460 (1992); (2) as direct oncolytic agents by using viruses that have been genetically modified to lose their pathogenic features, Martuza, R. L. , et al.,
Science,
252:854-856 (1991); or (3) as agents to selectively damage malignant cells using viruses which have been genetic engineered for this purpose, Bischoff, J. R. , et al.,
Science,
274:373-376 (1996).
Examples for the use of viruses against malignant gliomas include the following.
Herpes Simplex Virus dlsptk (HSVdlsptk), is a thymidine kinase (TK)-negative mutant of HSV. This virus is attenuated for neurovirulence because of a 360-base-pair deletion in the TK gene, the product of which is necessary for normal viral replication. It has been found that HSVdlsptk retains propagation potential in rapidly dividing malignant cells, causing cell lysis and death. Unfortunately, all defective herpes viruses with attenuated neuropathogenicity have been linked with serious symptoms of encephalitis in experimental animals. Wood, M. J. A., et al.,
Gene Therapy,
1:283-291 (1994). For example, in mice infected intracerebrally with HSVdlsptk, the LD
50
IC
(intracranial administration) is 10
6
pfu, a rather low dose. This limits the use of this mutant HSV. Markert, J. M., et al.,
Neurosurgery,
32:597-603 (1993). Other mutants of HSV have been proposed and tested. Nevertheless, death from viral encephalitis remains a problem. Mineta T. , et al.,
Nature Medicine,
1:938-943 (1995); Andreansky, S. , et al.,
Cancer Res.,
57:1502-1509 (1997).
Another proposal is to use retroviruses engineered to contain the HSV tk gene to express thymidine kinase which causes in vivo phosphorylation of nucleoside analogs, such as gancyclovir or acyclovir, blocking the replication of DNA and selectively killing the dividing cell. Izquierdo, M., et al.,
Gene Therapy,
2:66-69 (1995) reported the use of Moloney Murine Leukemia Virus (MOMLV) engineered with an insertion of the HSV tk gene with its own promoter. Follow-up of patients with glioblastomas that were treated with intraneoplastic inoculations of therapeutic retroviruses by MRI revealed shrinkage of tumors with no apparent short-term side effects. However, the experimental therapy had no effect on short-term or long-term survival of affected patients. Retroviral therapy is typically associated with the danger of serious long-term side effects (e.g. insertional mutagenesis).
Chen, S. H., et al.,
PNAS,
USA, 91:3054-3057 (1994) reported the direct injection of a recombinant into experimentally induced gliomas in athymic mice. ADV/RSV-TK is an adenovirus containing the HSV-tk gene under transcriptional control of the rous sarcoma virus long terminal repeat, followed by treatment with gancyclovir. The treatment caused tumor necrosis without apparent involvement of the cellular immune response. The treated animals survived >50 days after tumor inoculation as contrasted with control tumor inoculated animals all of which died after 23 days. However, further long-term toxicity testing of neuronal, glial and endothelial cells is necessary to assess the potential of genetically engineered retroviruses for the treatment of cancers.
Recently, a novel strategy to use human pathogenic viruses for the treatment of malignant disease was introduced. Adenovirus engineered to selectively replicate within and destroy malignant cells expressing a modified p53 tumor suppressor offers an opportunity to target malignant cells without causing unwanted side effects due to virus propagation at extratumoral sites. Bischoff, J. R., et al., supra.
Similar systems have been developed to target malignancies of the upper airways, tumors that originate within the tissue naturally susceptible to adenovirus infection and that are easy accessible. However, Glioblastoma multiforme, highly malignant tumors composed out of widely heterogeneous cell types (hence the denomination multlforme) are characterized by exceedingly variable genotypes and are unlikely to respond to oncolytic virus systems directed against homogeneous tumors with uniform genetic abnormalities.
The Cells of the Central Nervous System
It is important to recognize that there are two classes of cells in the brain, the neural cells (neurons) and the neuroglia cells (glia). Neurons process information received from the peripheral receptors giving rise to perception and memory. Motor commands are issued and transmitted also by means of neurons to the various muscles of the body. There are nine times more glial cells than neurons. The glial cells have mult

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