Suppression of cancer

Abstract

The present invention relates to polypeptides for use in suppressing cancer and cancer disorders. The treatment employs use of a non-cytotoxic protease, which is targeted to the cancer cell, and, when so delivered, the protease is internalised and inhibits secretion from the cancer cell.

1 . A polypeptide, for use in suppressing a cancer cell that expresses a SNARE protein responsible for secretion from said cancer cell, wherein the polypeptide comprises:
 (i) a non-cytotoxic protease, which protease is capable of cleaving said SNARE protein expressed in said cancer cell;   (ii) a Targeting Moiety (TM) that is capable of binding to a Binding Site on a cancer cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and   (iii) a translocation domain that is capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell;   wherein the polypeptide lacks the natural binding function of a clostridial neurotoxin H CC  domain which enables the clostridial neurotoxin to bind to nerve terminals at the neuromuscular junction;   with the proviso that the cancer cell is not a neuroendocrine tumour cell; and   with the proviso that the polypeptide is not a clostridial neurotoxin (holotoxin).   
     
     
         2 . The polypeptide for according to  claim 1 , wherein the cancer is selected from the group consisting of: lung cancer, renal cancer, brain cancer, breast cancer, pancreatic cancer, colorectal cancer, adrenal cancer, oesophageal cancer, lymphoma, B-cell lymphoma, mantle cell lymphoma, leukaemia, multiple myelona, acute leukaemia, bladder cancer, bone cancer, bowel cancer, cervical cancer, chronic lymphocytic leukaemia, Hodgkin's lymphoma, liver cancer, skin cancer, oropharyngeal cancer, myeloma, prostate cancer, prostate soft tissue sarcoma, gastric cancer, testicular cancer, uterine cancer, and Kaposi sarcoma. 
     
     
         3 . The polypeptide according to  claim 1 , wherein the cancer cell is selected from the group consisting of: a lung cancer cell, a renal cancer cell, a brain cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an adrenal cancer cell, an oesophageal cancer cell, a lymphoma cancer cell, a B-cell lymphoma cancer cell, a mantle cell lymphoma cancer cell, a leukaemia cancer cell, a multiple myeloma cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia, cell, a Hodgkin's lymphoma cell, a liver cancer cell, a melanoma skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a prostate cancer cell, a soft tissue sarcoma cell, a gastric cancer cell, a testicular cancer cell, a uterine cancer cell and a Kaposi sarcoma cancer cell. 
     
     
         4 . The polypeptide of  claim 1 , wherein the TM binds to a receptor selected from the group consisting of: an ErbE receptor; an EGF receptor; a growth hormone-releasing hormone (GHRH) receptor; an insulin-like growth factor receptor (IGFR); a gastrin-releasing peptide (GRP) receptor; a bombesin peptide (BB) receptor; a growth hormone (GH) receptor; a vascular endothelial growth factor (VEGF) receptor; an acetylcholine (ACH) receptor; a somatostatin (SST) receptor; a cortistatin (CST) receptor; a chemokine (C-X-C motif) receptor such as CXCR4; a neuropilin receptor; a gonadotropin-releasing hormone (GnRH) receptor; a VIP-glucagon-GRF-secretin superfamily receptor; a PAC receptor; a VPAC receptor; a FLT receptor; a CHRN receptor; an EPH receptor; an EPHA receptor; an EFN receptor; a DLK1 receptor, a DLL3 receptor, a FGF receptor; a JAG receptor; a LIF receptor; a NMB receptor; a NOTCH receptor; a PDGF receptor; a c-kit receptor; a TGF receptor; an endothelin receptor; a chemokine receptor; and an angiopoietin receptor. 
     
     
         5 . The polypeptide of  claim 1 , wherein the TM is selected from the group consisting of: an Erb B peptide, an EGF peptide, an adrenomedullin (ADM) peptide, an AM peptide, an AMF peptide, an amphiregulin peptide, an APRIL peptide, an artemin peptide, a betaceiluhn peptide, a bombesin peptide, a CTR peptide, an endothelin peptide, an erythropoietin peptide, a FGF peptide, a FSH peptide, a gastrin peptide, a gastrin releasing peptide (GRP), a GDNF peptide, a ghrelin peptide, a GHRH peptide, a G-CSF peptide, a growth hormone peptide, a HB-EGF peptide, a hepatocyte growth factor (HGF) peptide, an IL peptide, a keratinocyte growth factor peptide, a leptin peptide, a LIF peptide, an alpha-melanotropin peptide, a MGSA/GRO peptide, a NRG peptide, an oxytocin peptide, an osteopontin (OPN) peptide, a neuregulin-1 peptide, a VIP or PACAP peptide, a PDGF peptide, a prolactin peptide, a SCF peptide, a somatostatin (SST) peptide, a cortistatin (CST) peptide, a TNF peptide, a TGF peptide, a VEGF peptide, a vasopressin peptide, an angiopoietin peptide, a B-CLL peptide, a BCGF-12KD peptide, a BAFF peptide, a galanin peptide, a GDNF peptide, a GnRH peptide, an IGF-II peptide, a LH peptide, a neurotrophin peptide, a substance P peptide, a TGF-alpha peptide; an IGF peptide, an angiopoietin peptide, a CXC peptide, a CCL peptide, or an antibody or antibody fragment that binds to a receptor as defined in  claim 4 . 
     
     
         6 . The polypeptide of  claim 1 , wherein the non-cytotoxic protease comprises a clostridial neurotoxin L-chain or an IgA protease. 
     
     
         7 . The polypeptide of  claim 1 , wherein the translocation domain comprises a clostridial neurotoxin translocation domain. 
     
     
         8 . A polypeptide comprising:
 (i) a non-cytotoxic: protease, which protease is capable of cleaving a SNARE protein expressed in a cancer cell, wherein said SNARE protein is responsible for secretion from the cancer cell;   (ii) a Targeting Moiety (TM) that is capable of binding to a Binding Site on the cancer cell, which Binding Site is capable of undergoing endocytosis to be incorporated into an endosome within the cancer cell; and   (iii) a translocation domain that is capable of translocating the protease from within an endosome, across the endosomal membrane and into the cytosol of the cancer cell;   wherein the polypeptide lacks the natural binding function of a clostridial neurotoxin H CC  domain which enables the clostridial neurotoxin to bind to nerve terminals at the neuromuscular junction;   with the proviso that the cancer cell is not a neuroendocrine tumour cell; and   with the proviso that the polypeptide is not a clostridial neurotoxin (holotoxin).   
     
     
         9 . The polypeptide according to  claim 8 , wherein the TM binds to a receptor selected from the group consisting of: an ErbB receptor; an EGF receptor; a growth hormone-releasing hormone (GHRH) receptor; an insulin-like growth factor (IGF) receptor; a gastrin-releasing peptide (GRP) receptor; a bombesin peptide (BB) receptor a growth hormone (GH) receptor; a vascular endothelial growth factor (VFGF) receptor; an acetylcholine (ACH) receptor; a somatostatin (SST) receptor; a cortistatin (CST) receptor; a chemokine (C-X-C motif) receptor; CXCR4; a neuropilin receptor; a gonadotropin-releasing hormone (GnRH) receptor; a VIP-glucagon-GRF-secretin superfamily receptor; a PAC receptor; a VPAC receptor; a FLT receptor; a CHRN receptor; an EPH receptor; an EPHA receptor; an EFN receptor; a DLK1 receptor, a DLL3 receptor, a FGF receptor; a JAG receptor; a LIF receptor; a NMB receptor; a NOTCH receptor; a PDGF receptor; a c-kit receptor; a TGF receptor; an endothelin receptor; a chemokine receptor; and an angiopoietin receptor. 
     
     
         10 . The polypeptide according to  claim 8 , wherein the TM is selected from the group consisting of: an ErbB peptide, an EGF peptide, an adrenomedullin (ADM) peptide, an AM peptide, an autocrine motility factor (AMF) peptide, an amphiregulin peptide, an APRIL peptide, an artemin peptide, autocrine motility factor (AMF) peptide, a betacellulin peptide, a bombesin peptide, a CTR peptide, an endothelin peptide, an erythropoietin peptide, a FGF peptide, a FSH peptide, a gastrin peptide, a gastrin releasing peptide (GRP), a GDNF peptide, a ghrelin peptide, a GHRH peptide, a G-CSF peptide, a growth hormone peptide, a HB-EGF peptide, a hepatocyte growth factor (HGF) peptide, an IL peptide, a keratinocyte growth factor peptide, a leptin peptide, a LIF peptide, an aipha-melanotropin peptide, a MGSA/GRO peptide, a NRG peptide, an oxytocin peptide, an osteopontin (OPN) peptide, a neureguiin-1 peptide, a VIP peptide, a PACAP peptide, a PDGF peptide, a prolactin peptide, a SCF peptide, a somatostatin (SST) peptide, a cortistatin (CST) peptide, a TNF peptide, a TGF peptide, a VEGF peptide, a vasopressin peptide, an angiopoietin peptide, a B-CLL peptide, a BCGF-12KD peptide, a BAFF peptide, a galanin peptide, a GDNF peptide, a GnRH peptide, au IGF-II peptide, a LH peptide, a NT peptide, a substance-P peptide, a TGF-alpha peptide; an IGF peptide, an angiopoietin peptide, a CXC peptide, a CCL peptide, and an antibody or antibody fragment that binds to a receptor as defined in  claim 9 . 
     
     
         11 . The polypeptide according to  claim 8 , wherein the non-cytotoxic protease comprises a clostridial neurotoxin protease or an IgA protease. 
     
     
         12 . The polypeptide according to  claim 8 , wherein the translocation domain comprises a clostridial neurotoxin translocation domain. 
     
     
         13 . The polypeptide according to  claim 8 , wherein said polypeptide comprises au amino acid sequence having at least 90-92%, or at least 95-97%, or at least 98-99% sequence identity to any one of SEQ ID NOs: 7, 10, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, or 44. 
     
     
         14 . A nucleic acid encoding a polypeptide according to  claim 8 . 
     
     
         15 . A nucleic acid encoding a polypeptide according to  claim 8 , wherein said nucleic acid comprises a nucleic acid sequence haying at least 90-94%, or at least 95-97%, or at least 98-99% sequence identity to any one of SEQ ID NOs: 6, or 9. 
     
     
         16 . A method of suppressing a cancer in a patient, comprising administering to the patient an effective amount of a. polypeptide according to  claim 8 . 
     
     
         17 . The method according to  claim 16 , for suppressing a cancer selected from the group consisting of: lung cancer, renal cancer, brain cancer, breast cancer, pancreatic cancer, colorectal cancer, colon cancer, rectal cancer, adrenal cancer, oesophageal cancer, lymphoma, B-cell lymphoma, mantle cell lymphoma, leukaemia, multiple myeloma, acute leukaemia, bladder cancer, bone cancer, bowel cancer, cervical cancer, chronic lymphocytic leukaemia, Hodgkin's lymphoma, liver cancer, skin cancer, oropharyngeal cancer, myeloma, prostate cancer, prostate soft tissue sarcoma, gastric cancer, testicular cancer, uterine cancer and Kaposi sarcoma. 
     
     
         18 . The method according to  claim 16 , for suppressing secretion from a cancer cell selected from the group consisting of: a lung cancer cell, a renal cancer cell, a brain cancer cell, a breast cancer cell, a pancreatic cancer cell, a colorectal cancer cell, an adrenal cancer cell, an oesophageal cancer cell, a lymphoma cancer cell, a B-cell lymphoma cancer cell, a mantle cell lymphoma cancer cell, a leukaemia cancer cell, a multiple myeloma cancer cell, an acute leukaemia cancer cell, a bladder cancer cell, a bone cancer cell, a bowel cancer cell, a cervical cancer cell, a chronic lymphocytic leukaemia cell, a Hodgkin's lymphoma cell, a liver cancer cell, a skin cancer cell, an oropharyngeal cancer cell, a myeloma cell, a prostate cancer cell, a soft tissue sarcoma cell, a gastric cancer cell, a testicular cancer cell, a uterine cancer cell and a Kaposi sarcoma cell. 
     
     
         19 . A method of suppressing a cancer in a patient, comprising administering to the patient an effective amount of a nucleic acid according to  claim 14 . 
     
     
         20 . The polypeptide according to  claim 9 , wherein the TM is selected from the group consisting of: an ErbB peptide, an EGF peptide, an adrenomedullin (ADM) peptide, an AM peptide, an autocrine motility factor (AMF) peptide, an amphiregulin peptide, an APRIL peptide, an artemin peptide, an autocrine motility factor (AMF) peptide, a betacellulin peptide, a bombesin peptide, a CTR peptide, an endothelin peptide, an erythropoietin peptide, a FGF peptide, a FSH peptide, a gastrin peptide, a gastrin releasing peptide (GRP), a GDNF peptide, a ghrelin peptide, a GHRH peptide, a G-CSF peptide, a growth hormone peptide, a HB-EGF peptide, a hepatocyte growth factor (HGF) peptide, an IL peptide, a keratinocyte growth factor peptide, a leptin peptide, a LIF peptide, an alpha-melanotropin peptide, a MGSA/GRO peptide, a NRG peptide, an oxytocin peptide, an osteopontin (OPN) peptide, a neuregulin-1 peptide, a VIP peptide, a PACAP peptide, a PDGF peptide, a prolactin peptide, a SCF peptide, a somatostatin (SST) peptide, a cortistatin (CST) peptide, a TNF peptide, a TGF peptide, a VEGF peptide, a vasopressin peptide, an angiopoietin peptide, a B-CLL peptide, a BCGF-12KD peptide, a BAFF peptide, a galanin peptide, a GDNF peptide, a GnRH peptide, an IGF-II peptide, a LH peptide, a NT peptide, a substance-P peptide, a TGF-alpha peptide; an IGF peptide, an angiopoietin peptide, a CXC peptide, a CCL peptide, and an antibody or antibody fragment that binds to a receptor as defined in  claim 9 .