PLASMA CELL DISORDERS

Use of NOTCH Pathway Interfering Agents for Treatment of Plasma Cell Disorders

U.S. Patent Number: 7,282,203
Issue Date: October 16, 2007

Summary: The invention provides a method for reducing the severity of, or treatment of, plasma cell disorders. The method comprises the step of administering to an individual afflicted with a plasma cell disorder, a composition comprising an antibody directed to the extracellular portion of NOTCH or to JAG2.

Detail: The invention provides a method for novel therapeutic approaches to the treatment of plasma cell disorders. The method comprises the use of agents which interfere with the NOTCH pathway.

In one embodiment, the invention provides a method for reducing the severity of plasma cell disorders or treatment of plasma cell disorders comprising the steps of administering antibodies to NOTCH protein, particularly the extracellular portion of the NOTCH1 protein. In another embodiment, the invention provides a method for reducing the severity of plasma cell disorders or treatment of plasma cell disorders comprising the steps of administering antibodies to JAG2 protein. The JAG2 protein is a surface protein considered to be on malignant plasma cells.

In another embodiment, the invention provides a method for reducing the severity of plasma cell disorders or treating plasma cell disorders comprising the steps of administering antibodies to NOTCH protein and/or antibodies to the JAG2 protein in combination with a cytotoxic agent. An example of a useful chemotherapeutic agent is doxorubicin.

Method for Inducing Apoptosis of Primary Central Nervous System B Cell Lymphomas

U.S. Patent Number: 6,042,826
Issue Date: March 28, 2000

Summary: A method for treating a primary central nervous system lymphoma in an individual relates to administering intrathecally or intralesionally a therapeutically effective amount of a Fas-cross-linking composition thereby inducing the lymphoma cells to undergo Fas-mediated cytotoxicity. The Fas-cross-linking composition may be an agonist anti-human Fas monoclonal antibody or fragments thereof, soluble Fas-ligand (Fas-L), and a combination thereof. In another embodiment, the lymphoma is pretreated with a composition that enhances Fas-mediated cytotoxicity induced by a Fas-crosslinking composition, followed by treatment with the Fas-cross-linking composition.

Detail: Accordingly, it is a primary object of the invention to provide methods for treating individuals having PCNSL. It is another object of the invention to provide a method for treating individuals having PCNSL, wherein the method offers a mechanism for achieving better therapeutic efficacy than the current standard therapy for PCNSL. It is another object of the invention to provide a method for treating individuals having PCNSL, wherein the method offers a mechanism for achieving better therapeutic efficacy than the current standard therapy for PCNSL, and further offers a mechanism to minimize toxicity to the treated individual.

It is a further object of the invention to provide a method for treating individuals having PCNSL, wherein the method offers a mechanism for achieving better therapeutic efficacy than the current standard therapy for PCNSL, and further offers a mechanism to minimize toxicity to the treated individual, wherein the method is facilitated by Fas-mediated cytotoxicity of the malignant tumor. The foregoing objects are based on a novel discovery that primary central nervous system B cell lymphomas, unexpectedly, express significant amounts of Fas on their cell-surface, and further, that ligation (cross-linking) of the receptor on these Fas-bearing tumor cells leads to rapid programmed cell death of the treated tumor. In another embodiment of the present invention, the method comprises combination therapy involving the use of one or more compositions for ligation of Fas expressed by primary central nervous system B cell lymphomas in conjunction with one or more compositions which potentiates or enhances Fas–mediated cytoxitcity of the treated tumor, such as by upregulating the cell-surface expression of Fas by the tumor.

SOLID TUMORS

Method for Treating Hyperproliferative Tissue in a Mammal

U.S. Patent Number: 6,495,585
Issue Date: December 17, 2002

Summary: A novel method for treating undesired hyperproliferative tissue in a mammal. The method includes the steps of: injecting the mammal with a photodynamic compound having a selective uptake in the hyperproliferative tissue and which is activated at a particular light frequency; injecting the mammal with a xanthenone-4-acetic acid or a Group I metal, Group II metal or quaternary salt thereof near the time of maximum uptake of the photodynamic compound in the hyperproliferative tissue; and exposing the hyperproliferative tissue to light at the particular frequency that activates the photodynamic compound. The method of the invention causes necrosis of the hyperproliferative tissue to an extent greater than can be obtained by either the photodynamic compound or xanthenone-4-acetic acid alone. Further and surprisingly the method enhances immune response of the mammal to the hyperproliferative tissue even after the photodynamic compound and xanthenone-4-acetic acid are no longer present in the mammal.

Detail: In accordance with the invention a novel method is provided for treating undesired hyperproliferative tissue in a mammal. The method includes the steps of: injecting the mammal with a photodynamic compound having a selective uptake in the hyperproliferative tissue and which is activated at a particular light frequency; injecting the mammal with a xanthenone-4-acetic acid or a Group I metal, Group II metal or quaternary salt thereof near the time of maximum uptake of the photodynamic compound in the hyperproliferative tissue; and exposing the hyperproliferative tissue to light at the particular frequency that activates the photodynamic compound. The method of the invention causes necrosis of the hyperproliferative tissue to an extent greater than can be obtained by either the photodynamic compound or xanthenone-4-acetic acid alone. Further and surprisingly the method enhances immune response of the mammal to the hyperproliferative tissue even after the photodynamic compound and xanthenone-4-acetic acid are no longer present in the mammal.

p53 as Protein and Antibody Therefor

U.S. Patent Number: 7,238,523 (CIP); 6,965,009 (DIV)
Issue Date: July 3, 2007 (CIP); November 15, 2005 (DIV)

Summary: The invention comprises plasmids and viral vectors containing an animal p53as cDNA sequence. A portion of the p53as sequence may be identified to a position of wild type p53 gene from the same animal. In preferred embodiments, the p53as is mouse or human p53as. A preferred viral vector is baculovirus vector. The invention further includes antibodies both polyclonal and monoclonal, to p53as and to at least a portion of human p53 intron 10 sequence encoding SLRPFKALVREKGHRPSSHSC (SEQ. I.D. NO. 1) which is related to p53as sequences and plasmids and viral vectors containing such sequences. All of the above find utility in studying p53 and p53as and their relative expressions which is believed important for detection and control of malignant cells and their susceptibility to treatment agents. The antibodies can detect the presence of p53as and related sequences and when injected into cells could cause cell cycle arrest and the plasmids and viral vectors, with appropriate promoters, can cause expression of the p53as and p53 intron 10 sequences which can affect cell growth and perhaps arrest certain malignancies.

Detail: p53as protein exhibited the antibody binding properties of wild type p53 protein, PAb246+, PAb240-, but lacked the C-terminal epitope reactive with PAb421. p53as protein translated in vitro was activated for binding to a p53 DNA binding sequence. The major p53 protein, in contrast, required activation for DNA binding (by monoclonal antibody PAb421). There appears to be a direct interaction between p53as and p53 proteins which influenced the composition of the DNA binding complex and the magnitude of DNA binding. Because cotranslated p53 protein inactivated p53as protein for DNA binding, and because two bands were super-shifted by PAb421 in lysates containing p53as and p53 cotranslated proteins, compared to one band in lysate containing p53 alone. These results could be explained by binding to DNA of p53as and p53/p53as heterooligomers, in addition to the binding of p53. The significance of these findings is that they are consistent with a functional role for p53as protein in cells, which may, at least in part, be distinct from the function(s) of the major p53 protein form. Considering that vectors and plasmids containing the p53 gene are being tested for applications in gene therapy, and considering the results herein that p53as is active for binding to a p53 binding sequence and that p53as interacts with p53, resulting in altered DNA binding, plasmids and vectors for the expression of mouse and human p53as in cells and for uses in gene therapy in humans are claimed herein.

GD2 Peptide Mimics

U.S. Patent Number: 6,939,948
Issue Date: September 6, 2005

Summary: The invention provides peptide mimics for GD2 ganglioside. The peptide mimics were identified by panning phage display peptide libraries with an anti-GD2 monoclonal antibody. The identified peptide mimics can be used as immunogens for cancer therapy such as for melanoma and neuroblastoma.

Detail: The invention provides methods of using compositions which elicit an immune response against a tumor associated antigen that is not normally immunogenic. The method comprises using peptide mimics to a ganglioside, GD2, to elicit an immune response. Accordingly, in one aspect, the invention provides methods for identifying peptide mimics. The method comprises the steps of screening phage display peptide libraries with antibodies to GD2. The identified peptides are then tested for their ability to elicit an immure response and the ability of those antibodies to against GD2 bearing cells. In another embodiment, the invention provides a method for eliciting an immune response in patients with GD2 positive tumors. The method comprises administering a composition effective in stimulating a specific immunological response against the GD2 antigen. These composition(s) comprise a peptide that shares immunological characteristics of GD2. While a detectable immunological response is likely to be beneficial, efficacy can also be deduced by an improvement in symptoms or control of growth of the tumor.

Other embodiments include include methods for treating GD2 bearing tumors in an individual by eliciting an antiGD2 immunological response in the subject. The immunological response can be elicited using any of the peptide mimics to the GD2. Still other embodiment include preparing a composition for use in the generation of an immune response and in the treatment of tumors bearing GD2. The composition comprises the peptide mimics disclosed herein.

GD3 Peptide Mimics

U.S. Patent Number: 6,998,237
Issue Date: February 14, 2006

Summary: The invention provides peptide mimics for GD3 ganglioside. The peptide mimics were identified by panning phage display peptide libraries with an anti-GD3 monoclonal antibody. The peptide mimics inhibit the binding of an anti-GD3 antibody to GD3 positive cells and also elicit antibodies which can bind to GD3 positive cells. The identified peptide mimics can be used as immunogens for cancer therapy.

Detail: The invention provides peptide mimics of the ganglioside GD3 and a method for producing same. This invention also provides a method of using the peptides to elicit an immune response against a tumor associated antigen that is not normally immunogenic. Accordingly, in one aspect, the invention provides methods for identifying peptide mimics. The method comprises the steps of screening phage display peptide libraries with antibodies to GD3. The identified peptides are then tested for their ability to elicit an immure response and the reactivity of those antibodies against GD3 bearing cells. In another aspect, the invention provides a method for eliciting an immune response in patients with GD3 positive tumors. The method comprises administering a composition effective in stimulating a specific immunological response against the GD3 antigen. These composition(s) comprise a peptide that shares immunological characteristics of GD3. While a detectable immunological response is likely to be beneficial, efficacy can also be deduced by an improvement in symptoms or control of growth of the tumor.

Other aspects include methods for treating GD3 bearing tumors in an individual by eliciting an anti GD3 immunological response in the subject. The immunological response can be elicited using any of the peptide mimics to the GD3. Still other embodiment include preparing a composition for use in the generation of an immune response and in the treatment of tumors bearing GD3. The composition comprises the peptide mimics disclosed herein.

Method for Inhibition of Angiogenesis and Vasculogenesis

U.S. Patent Number: 6,977,271
Issue Date: December 20, 2005

Summary: The invention provides a method for inhibiting or preventing angiogenesis in animals and humans comprising the steps of administering to the animal or human an effective amount of one or more isomers of conjugated linoleic acid ("CLA"). The CLA may be administered in the form of a free fatty acid, an ester, or as a salt, in a pill, or as a component of a natural or prepared food product.

Detail: The invention provides a method for inhibition of angiogenesis in an individual diagnosed with a solid tumor. The method comprises administering orally a therapeutically effective dose of one or more isomers of conjugated linoleic acid (CLA) such that angiogenesis in the tumor is reduced following administration of the CLA isomer or isomers. The invention also provides a method for prevention of the occurrence or severity of solid tumors comprising identifying individuals with a high risk of developing solid tumors (such as breast cancer) and administering one or more isomers of CLA.

Method for Identifying and Designing Immunogenic Peptides

U.S. Patent Number: 7,348,153
Issue Date: December 20, 2005

Summary: The invention provides a method for identifying peptides for use in increasing a cytotoxic T lymphocytes (CTL) response to an antigen. The method comprises the steps of comparing the amino acid sequence of the VH and/or VL portions of an anti-idiotypic monoclonal antibody to the amino acid sequence of an antigen to identify regions of homology between the Ab2 and the antigen, and to further identify an HLA binding motif in a homologous region. The identified homologous region which comprises an HLA binding motif defines a peptide sequence that is useful for stimulating a CTL response. Also provided are peptides identified by the method, and a method of using the peptides to stimulate a CTL response in an individual.

Detail: The invention provides a method for identifying peptides for use in immunotherapeutic approaches. The peptides are identified as short regions (8-20 amino acid) of an anti-idiotypic antibody having homology with a region of the corresponding antigen and further comprising one or more HLA binding motif(s). These peptides have the ability to stimulate a cytotoxic T lymphocyte (CTL) response. The peptide regions of the ant-id antibodies may be modified to further enhance the immunogenic response.In one embodiment, the method comprises obtaining the amino acid sequences of the VH and/or VL portions of the anti-idiotypic antibody (Ab2), obtaining the sequence of the antigen, comparing the sequences to identify candidate homologous regions within the Ab2 (i.e., regions that show homology to regions of the corresponding antigen), and further identifying those candidate homologous regions of Ab2 which also comprise HLA binding motifs. These peptides are then tested for their ability to stimulate a CTL response. Preferred peptides are those which stimulate a greater CTL response than the corresponding antigen or a peptide derived from the antigen. In another embodiment, the invention also provides peptides identified by the method. In this regard, the peptide sequences are homologous to a portion of Ab2 and a portion of the antigen, and further comprise an HLA binding motif.In another embodiment, the method further comprises modifying the peptide sequences identified as homologous regions of Ab2 to further enhance their ability to stimulate a CTL response. This invention also provides a method for using the peptides identified by the method in generating an increased immunogenic response in an individual against the tumor antigen. The method comprises administering to the individual an amount of peptide which has homology to a portion of the antigen and to a portion of an Ab2 which mimics an epitope on the antigen, and wherein the peptide comprises an HLA binding motif, effective to stimulate a CTL mediated immune response against the antigen.

Method of Treating Solid Tumors and Leukemias Using Combination Therapy of Vitamin D and Anti-Metabolic Nucleoside Analogs

U.S. Patent: Pending
Application Number: 11/093,123

Summary: The invention relates to a method of inhibiting solid tumor cell or leukemia cell proliferation by first administering to a solid tumor cell or leukemia cell either vitamin D or a derivative thereof and subsequently administering at least one anti-metabolic nucleoside analog to the solid tumor cell or the leukemia cell. Also disclosed is a method of treating a cancerous condition. Methods of down regulating a p-AKt survival signaling pathway and modulating activity of a pro-apoptotic caspases is also disclosed.

Detail: The invention relates to a method of inhibiting solid tumor cell or leukemia cell proliferation. This method involves first administering to a solid tumor cell or a leukemia cell either vitamin D or a derivative thereof and subsequently administering at least one antimetabolic nucleoside analog to the solid tumor cell or the leukemia cell. The solid tumor cell or the leukemia cell is susceptible to the first administering and the subsequent administering and proliferation thereof is inhibited. A second aspect of the invention relates to a method of treating a cancerous condition. This method involves first administering to a patient diagnosed with a cancerous condition selected from the group consisting of a solid tumor or a leukemia, either vitamin D or a derivative thereof and subsequently administering to the patient at least one anti-metabolic nucleoside analog. The cancer cells associated with the cancerous condition are susceptible to the first administering and the subsequent administering and progression of the cancerous condition is inhibited. A third aspect of the invention relates to a method of downregulating a p-Akt survival signaling pathway in a cell, particularly a cancer cell. This method involves exposing a cell exhibiting a p-Akt survival signaling pathway to a combination of (i) vitamin D or a derivative thereof and (ii) at least one anti-metabolic nucleoside analog. The exposing step reduces the activity of the p-Akt survival signaling pathway in the cell. A fourth aspect of the invention relates to a method of modulating activity of a pro-apoptotic caspase. This method involves exposing a cell to a combination of (i) vitamin D or a derivative thereof and (ii) at least one anti-metabolic nucleoside analog. The exposing step enhances the activity of a pro-apoptotic caspase in the cell. The invention demonstrates the efficacy of vitamin D (or its derivatives) in a combination therapy with anti-metabolic nucleoside drugs, particularly drugs that induce pro-apoptotic caspase activity such as the pancreatic cancer drug gemcitabine. As demonstrated in the examples, the combination of gemcitabine and calcitriol were able to synergistically activate pro-apoptotic caspase activity while decreasing anti-apoptotic (pro-survival) p-Akt pathways in an in vivo pancreatic cancer model. The demonstration of effective therapies for pancreatic cancer is, in particular, quite important because pancreatic cancer generally has a poor prognosis. Very often, diagnosis occurs only when the disease is quite advanced. Consequently, of those diagnosed, about twenty percent survive one year and fewer than five percent survive five years. Thus, a preferred embodiment of the present invention provides a significant improvement in pancreatic cancer therapy.

5-Amino-4-Imidazolecarboxamide Riboside and Its Nucleobase as Potentiators of Antifolate Transport and Metabolism

U.S. Patent: Pending
Application Number: 11/327,872

Summary: The invention provides a method for increasing the efficacy of antifolates which act via inhibition of dihydrofolate reductase (DHFR). The method comprises the steps of administration of 5-amino-4-imidazolecarboxamide riboside (Z) or its base with the antifolate such that the targeted cells are exposed to both the antifolate and Z simultaneously. This results in increased influx of the antifolate. For MTX, accumulation of the more biologically active polyglutamate forms is also potentiated. This potentiation appears to be mediated by an effect on the RFC.

Detail: The invention provides a method for enhancing the uptake and efficacy of antifolates which act via inhibition of DHFR such as the 2,4 diaminopteridine antifolates methotrexate and aminopterin.The method is based on the unexpected observation that exogenous 5-amino-4-imidazolecarboxamide riboside (Z), a nucleoside precursor of (among others) the triphosphate ZTP, potentiates uptake of MTX and synthesis of MTX polyglutamate in cancer cells. Based on the data presented herein, it is considered that Z potentiates transport of antifolates via the RFC and the increased transport leads to increased synthesis of antifolate polyglutamates and consequently increased drug accumulation. Z was observed to enhance the growth inhibitory potency of MTX against cancer cells.Thus in one embodiment, this invention provides a method comprising the administration of Z or its base (i.e., 5-amino-4-imidazolecarboxamide) with an antifolate which acts via inhibition of the DHFR at concentrations at which the antifolate inhibits DHFR. The administration of Z or its base can be accomplished by any standard method, although systemic administration is preferred. Z has already been tested in clinical trials as a treatment for cardiac ischemia and is known to be nontoxic.In another embodiment, Z or its base and an antifolate which acts via inhibition of DHFR can be administered with a second antifolate(s) which primarily act via another mechanism such as inhibition of thymidylate synthase, inhibition of purine synthesis or other multi-targeted inhibition pathways. Administration of Z or its base with folate(s) which inhibit DHFR (with or without other folates) to enhance the efficacy of the folate(s) can be carried out for inhibiting the growth of cells as in various cancers as well as in other pathological conditions such as rheumatoid arthritis and psoriasis.

Acyl Homoserine Lactones for Inhibition of Cell Growth

U.S. Patent: Pending
Application Number: 11/399,933

Summary: The invention provides a method for inhibiting the growth of cancer cells using AHLs of the general formula CX-homoserine lactone where “X” represents a number of between 5 and 14 carbon atoms in the acyl chain of the AHL. The method comprises the step of administering to an individual an amount of an AHL effective to inhibit the growth of cancer cells. Also provided is a method for enhancing the effect of a chemotherapeutic agent comprising the step of administering to an individual the chemotherapeutic agent and an amount of an AHL effective to enhance the cancer cell growth inhibitory effect of the chemotherapeutic agent.

Detail: The invention is based on the discovery that human cancer cells which over-express rTS.beta. secrete one or more compounds capable of causing down regulation of TS expression, that the conditioned medium of these cells contains AHLs, that rTS.beta. is involved in the synthesis of AHLs, and that AHLs can downregulate TS. Based on these unexpected findings, the present invention provides a method for inhibiting the growth of cancer cells in an individual. The method comprises the step of administering to the individual a composition comprising an amount of an AHL effective to inhibit the growth of the cancer cells. It is demonstrated that the administration of AHLs can inhibit the growth of a variety of cancer cells, including human colorectal, lung, breast and prostate cancer cells. AHLs useful in the method of the invention are those of the L form and which have the general formula of CX-homoserine lactone, where “X” represents a number of between 5 and 14 carbon atoms in the acyl chain of the AHL. Certain positions of the AHLs can also be modified, such as by addition of an aromatic group to the acyl chain, without adversely affecting their capacity to inhibit cancer cell growth. It is preferred that the AHLs are 3-oxo-CX-homoserine lactones. Also provided is a method for enhancing the effect of a chemotherapeutic agent. The method comprises the step of administering to an individual a chemotherapeutic agent and an amount of an AHL effective to enhance the cancer cell growth inhibitory effect of the chemotherapeutic agent relative to administration of the chemotherapeutic agent alone. In particular, it is shown that AHLs can enhance the effect of chemotherapeutic agents known to predominantly target TS, as well as the effects of chemotherapeutic agents thought to act in a manner unrelated to TS expression, such as microtubule inhibitors. Moreover, AHLs can enhance the activity of microtubule inhibitors against cells which are known to be resistant to chemotherapeutic agents that affect TS.

Peptides for Stimulating an Immune Response Against Melanoma

U.S. Patent : Pending
Application Number: 11/487,877

Summary: Provided in the invention are recombinant peptides and a method for using the peptides in stimulating an immune response against human high molecular weight-melanoma associated antigen (HMW-MAA). The peptides were designed from the identification of regions of structural and amino acid sequence homology between HMW-MAA and the mouse anti-idiotypic monoclonal antibody MK2-23. The method comprises the step of administering to an individual a peptide of the invention in an amount effective to elicit an immune response against HMW-MAA.

Detail: The invention provides recombinant peptides for use in stimulating an immune response against melanoma. The peptides were designed from regions of structural and amino acid sequence homology identified herein between HMW-MAA and the mouse anti-idiotypic monoclonal antibody (anti-id mAb) MK2-23, which mimics an HMW-MAA epitope. In particular, X-ray crystallography analysis of the Fab' portion of MK2-23 was used to identify regions of the heavy and light chains of the MK2-23 anti-idiotypic antibody which displayed similar folding patterns as the region of the HMW-MAA comprising the epitope mimicked by MK2-23. These studies indicated that the complementarity determining region 3 (CDR3) of its heavy chain (also referred to herein as "H3") and the complimentarity determining region 1 (CDR1) of its light chain (also referred to herein as "L1") display partial amino acid sequence homology and a similar structural folding as a portion of the HMW-MAA protein that comprises the epitope which is mimicked by MK2-23. Based on these data, three peptides are provided--one each from the H3 and the L1 region of MK2-23 and one from the HMW-MAA.The invention also provides a method for using the peptides identified herein for stimulating an immune response in an individual against melanoma. The method comprises administering to the individual an amount of a composition comprising one or more peptides of the invention in an amount effective to stimulate an immune response against HMW-MAA.

Inhibition of Breast Carcinoma Stem Cell Growth and Metastasis

U.S. Patent: Pending
Application Number: 11/724,884

Summary: Disclosed is a method for inhibiting the growth of breast carcinoma stem cells that express High Molecular Weight -Melanoma Associated Antigen (HMW-MAA). The method comprises administering to an individual a composition comprising an antibody reactive with HMW-MAA or a fragment of such an antibody in an amount effective to inhibit the growth of the breast carcinoma cells. Also provided are methods for inhibiting metastasis of breast carcinomas and methods for identifying HMW-MAA+ breast cancer stem cells.

Detail: The invention provides a method for inhibiting the growth of breast carcinoma stem cells. The breast carcinoma stem cells express High Molecular Weight--Melanoma Associated Antigen (HMW-MAA). The method comprises administering to an individual a composition comprising an antibody reactive with HMW-MAA in an amount effective to inhibit the growth of the breast carcinoma stem cells. In another embodiment, a method is provided for inhibiting metastasis of a breast carcinoma where the breast carcinoma comprises HMW-MAA+ breast carcinoma stem cells. The method comprises administering to the individual a composition comprising an amount of an antibody reactive with HMW-MAA effective to inhibit metastasis of the breast carcinoma. In another embodiment, a method is provided for detection of HMW-MAA+ breast carcinoma stem cells. The method comprises administering to an individual, or contacting a biological sample obtained from the individual with, a combination of antibodies, where the combination of antibodies comprises an antibody directed HMW-MAA and at least one antibody directed to a breast cancer stem cell marker. Detecting the binding of both the HMWMAA antibody and the at least one breast cancer stem cell marker determines the presence of an HMA-MAA+ breast carcinoma stem cell. In particular embodiments, the antibody employed in practicing the invention can be the monoclonal antibody designated 225.28 and/or the monoclonal antibody designated 763.74.

Stress Protein Compositions and Methods for Prevention and Treatment of Cancer and Infectious Disease

U.S. Patent Number: 6,984,384; 7,378,096 (CIP)
Issue Date: January 10, 2006; May 27, 2008 (CIP)

Summary: Pharmaceutical compositions comprising a stress protein complex and related molecules encoding or cells presenting such a complex are provided. The stress protein complex comprises an hsp110 or grp170 polypeptide complexed with an immunogenic polypeptide. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of infectious disease. The invention further provides a method for producing T cells directed against a tumor cell or a M. tuberculosis- infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with a tumor or with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell, or with a tumor cell in a method for killing a tumor cell.

Detail: The invention provides a pharmaceutical composition comprising a stress protein complex. The stress protein complex comprises an hsp110 or grp170 polypeptide and an immunogenic polypeptide. In some embodiments, the hsp110 or grp170 polypeptide is complexed wit h the immunogenic polypeptide, for example, by non-covalent interaction or by covalent interaction, including a fusion protein. In some embodiments, the complex is derived from a tumor. In other embodiments, the complex is derived from cells infected with an infectious agent. The immunogenic polypeptide of the stress protein complex can be associated with a cancer or an infectious disease. The stress protein complex of the invention can further include additional stress polypeptides, including members of the hsp70, hsp90, grp78 and grp94 stress protein families. In one embodiment, the stress protein complex comprises hsp110 complexed with hsp70 and/or hsp25. The invention additionally provides a pharmaceutical composition comprising a first polynucleotide encoding an hsp 110 or a grp170 polypeptide and a second polynucleotide encoding an immunogenic polypeptide. In some embodiments involving first and second polynucleotides, the first polynucleotide is linked to the second polynucleotide. The pharmaceutical compositions of the invention can further comprise a physiologically acceptable carrier and/or an adjuvant. The efficacy of a pharmaceutical composition can further comprise GM-CSF-secreting cells. Alternatively, GM-CSF-secreting cells can be co-administered with a pharmaceutical composition of the invention, by administration before, during or after administration of the pharmaceutical composition. The use of GM-CSF-secreting cells enhances the efficacy of the pharmaceutical composition. In some embodiments, the complex is purified from a tumor or from cells infected with an infectious agent. In such embodiments, the stress polypeptide, as purified, is complexed with one or more immunogenic polypeptides. The binding of the stress polypeptide to the immunogenic polypeptide can be altered and/or enhanced by stress, such as by exposure to heat, anoxic and/or ischemic conditions, or proteotoxic stress. In particular, a stress protein complex of the invention can comprise a stress polypeptide complexed with an immunogenic polypeptide, wherein the complex has been heated. Such heating, particularly wherein the stress polypeptide comprises a heat-inducible stress protein, can increase the efficacy of the stress protein complex as a vaccine. Examples of heat-inducible stress proteins include, but are not limited to, hsp70 and hsp 110. In some embodiments, the immunogenic polypeptide is known. The immunogenic polypeptide is a known molecule, the immunogenic polypeptide can be provided in admixture with the stress polypeptide, or as a complex with the stress polypeptide. The hsp 110 or grp170 polypeptide can be complexed with the immunogenic polypeptide by non-covalent binding. Alternatively, the complex can comprise a fusion protein, wherein the stress polypeptide is linked to the immunogenic polypeptide. Examples of immunogenic polypeptides include, but are not limited to, antigens associated with cancer or infectious disease, such as the breast cancer antigen her2/neu or the Mycobacterium tuberculosis antigens Mtb8.4 and Mtb39. Where the immunogenic polypeptide is unknown, it can be obtained incidentally to the purification of the stress polypeptide from tissue of a subject having cancer or an infectious disease. Also provided is a pharmaceutical composition comprising an antigen-presenting cell (APC) modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide. Alternatively, the APC can be modified to present an immunogenic polypeptide obtained by purification of hsp 110 or grp170 from disease cells, including cancer cells and cells infected with an infectious agent. Preferably, the APC is a dendritic cell or a macrophage. The APC can be modified by various means including, but not limited to, peptide loading and transfection with a polynucleotide encoding an immunogenic polypeptide. The pharmaceutical compositions of the invention can be administered to a subject, thereby providing methods for inhibiting M. tuberculosis-infection, for inhibiting tumor growth, for inhibiting the development of a cancer, and for the treatment or prevention of cancer or infectious disease. The invention further provides a method for producing T cells directed against a tumor cell. The method comprises contacting a T cell with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell. Such T cells can be used in a method for killing a tumor cell, wherein the tumor cell is contacted with the T cell. Likewise, the invention provides a method for producing T cells directed against a M. tuberculosis-infected cell, wherein a T cell is contacted with an APC that is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the M. tuberculosis-infected cell. Included in the invention are T cells produced by this method and a pharmaceutical composition comprising such T cells. The T cells can be contacted with a M. tuberculosis-infected cell in a method for killing a M. tuberculosis-infected cell The T cells can be CD4+ or CD8+. The invention also provides a method for removing tumor cells from a biological sample. The method comprises contacting a biological sample with a T cell of the invention. In a preferred embodiment, the biological sample is blood or a fraction thereof. Also provided is a method for inhibiting tumor growth in a subject. The method comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp 110 or grp 170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate. The method further comprises administering to the subject an effective amount of the proliferated T cells, and thereby inhibiting tumor growth in the subject. In an alternative embodiment, the method for inhibiting tumor growth in a subject comprises incubating CD4+ and/or CD8+ T cells isolated from the subject with an antigen presenting cell (APC), wherein the APC is modified to present an hsp110 or grp170 polypeptide and an immunogenic polypeptide associated with the tumor cell such that T cells proliferate, cloning at least one proliferated cell, and administering to the patient an effective amount of the cloned T cells, thereby inhibiting tumor growth in the subject.