Sphingosine 1-Phosphate Antagonism

Abstract

Materials and Method for treating cancer and screening for anti-neoplastic agents are provided. These materials and methods can include sphingosine 1-phosphate antagonists that bind to sphingosine-1 phosphate receptor subtype 3. Antibodies and aptamers that selectively bind to an epitope in the extracellular loop between transmembrane domains two and three of sphingosine-1-phosphate receptor subtype 3 are provided.

1 - 73 . (canceled) 
     
     
         74 . An isolated sphingosine-1-phosphate (S1P) antagonist that selectively binds to an epitope in the extracellular loop between transmembrane domains two and three of sphingosine-1-phosphate receptor subtype 3 (S1P3), wherein the epitope optionally comprises the amino acid sequence KKTFSLSPTVWFLREG and wherein the S1P antagonist optionally comprises an antibody or epitope-binding antibody fragment, wherein the antibody or epitope-binding antibody fragment is a monoclonal antibody or fragment thereof, wherein the monoclonal antibody is optionally a chimeric antibody or a humanized antibody, wherein the monoclonal antibody is optionally produced from hybridoma EDDbeta7H9 or hybridoma EDDbeta7F6. 
     
     
         75 . An immunoconjugate, comprising a S1P antagonist according to  claim 74  conjugated to a functional agent, wherein the functional agent optionally is a cytotoxic agent optionally is selected from the group consisting of an aptamer (optionally a peptide or a nucleic acid, which nucleic acid may optionally comprise DNA and/or RNA), an antibiotic, a radioactive isotope, a nucleolytic enzyme, a toxin, and any combination thereof. 
     
     
         76 . A composition comprising a S1P antagonist according to  claim 74 , and optionally further comprising at least one pharmaceutically acceptable excipient. 
     
     
         77 . A composition according to  claim 76  that further comprises an anti-neoplastic agent, optionally: an anti-neoplastic agent that binds to an estrogen receptor or is an antimetabolite, taxane, anthracycline, progestin, megestrol, aromatase inhibitor, tyrosine kinase inhibitor, or epidermal growth factor inhibitor, any prodrug thereof, any salt thereof, and any combination thereof; is selected from the group consisting of tamoxifen, toremifene, raloxifene, clomiphene, any prodrug thereof, any salt thereof, and any combination thereof; is selected from the group consisting of methotrexate, capecitabine, cladribine, cytarabine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, thioguanine any prodrug thereof, any salt thereof, and any combination thereof; is selected from the group consisting of paclitaxel, docetaxel, any prodrug thereof, any salt thereof, and any combination thereof; is selected from the group consisting of doxorubicin, daunorubicin, idarubicin, epirubicin, any prodrug thereof, any salt thereof, and any combination thereof; is megestrol, a prodrug thereof, a salt thereof, or any combination thereof; is selected from the group consisting of methotrexate, capecitabine, cladribine, cytarabine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, thioguanine any prodrug thereof, any salt thereof, and any combination thereof; is selected from the group consisting of aminoglutethimide, anastrozole, letrozole, exemestane, any prodrug thereof, any salt thereof, and any combination thereof; and is selected from the group consisting of gefitinib, cetuximab, lapatinib, erlotinib, trastuzumab, any prodrug thereof, any salt thereof, and any combination thereof. 
     
     
         78 . A kit comprising an S1P antagonist according to  claim 74 , and optionally further comprising an anti-neoplastic agent, wherein the anti-neoplastic agent is optionally selected from the group consisting of an estrogen receptor binder, an antimetabolite, a taxane, an anthracycline, a progestin, a megestrol, an aromatase inhibitor, a tyrosine kinase inhibitor, or an epidermal growth factor inhibitor, any prodrug thereof, any salt thereof, and any combination thereof. 
     
     
         79 . A hybridoma that produces a S1P antagonist according to  claim 74  that is monoclonal antibody, wherein the hybridoma optionally is selected from the group consisting of EDDbeta7H9, EDDbeta7F6, and any combination thereof. 
     
     
         80 . A method selected from the group consisting of:
 a. a method of treating cancer, comprising administering an effective amount of a first anti-neoplastic agent comprising a S1P antagonist according to  claim 74  to a patient having or suspected of having a cancer, thereby treating cancer, optionally breast cancer, colorectal cancer, cervical cancer, endometrial cancer, ovarian cancer, prostate cancer, lung cancer, or glioblastoma, and wherein the method optionally further comprises administering an effective amount of a second anti-neoplastic agent, wherein the first anti-neoplastic agent enhances the efficacy of the second anti-neoplastic agent, optionally in an additive or synergistic manner;   b. a method for identifying a test agent that enhances the efficacy of an anti-neoplastic agent in antagonizing a neoplastic cell, comprising: (i) providing a first sample comprising a neoplastic cell; (ii) providing a second sample comprising a neoplastic cell; (iii) contacting the first and second samples with an anti-neoplastic agent; (iv) contacting the second sample with the text agent; (v) assaying the first and second samples for an anti-neoplastic effect, optionally cell stabilization, cell death, growth inhibition, cytoskeletal stabilization, and/or migration inhibition; and (vi) identifying whether the test agent is an enhancer based on the effect measured in the second sample compared to the effect measured in the first sample, wherein the anti-neoplastic agent is an S1P antagonist according to  claim 74 , and wherein the neoplastic cells are optionally selected from the group consisting of breast cancer cells, colorectal cancer cells, cervical cancer cells, endometrial cancer cells, ovarian cancer cells, prostate cancer cells, lung cancer cells, glioblastoma cells, and any combination thereof   c. a method of treating systemic inflammation, comprising administering an effective amount of a S1P antagonist according to  claim 74  to a subject in need thereof, thereby treating the subject's systemic inflammation;   d. a method of treating sepsis, including optionally sepsis with disseminated intravascular coagulation, comprising administering an effective amount of a S1P antagonist according to  claim 74  to a subject in need thereof, thereby treating sepsis;   e. a method of treating a viral hemorrhagic fever, comprising administering an effective amount of a S1P antagonist according to  claim 74  to a subject in need thereof, thereby treating the viral hemorrhagic fever; and   f. a method of preventing disseminated intravascular coagulation, comprising administering to a subject an amount of a S1P antagonist according to  claim 74  effective to prevent to disseminated intravascular coagulation in a subject, thereby preventing disseminated intravascular coagulation in the subject.