Therapeutic peptidomimetic macrocycles

Abstract

The present invention provides biologically active peptidomimetic macrocycles for the treatment of cell proliferative disorders such as cancer and immunoproliferative disease.

1 . A method of treating cancer in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle, wherein the cancer is selected from the group consisting of ovarian cancer, prostate cancer, renal cancer, breast cancer, pancreatic cancer, and Ph+ acute lymphocytic leukemia. 
     
     
         2 . The method of  claim 1 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         3 . The method of  claim 1 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         4 . The method of  claim 1 , wherein the peptidomimetic macrocycle is a BIM polypeptide. 
     
     
         5 . The method of  claim 4 , wherein an amino acid sequence of said BIM polypeptide is more than about 60% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         6 . The method of  claim 4 , wherein an amino acid sequence of said BIM polypeptide is more than about 80% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         7 . The method of  claim 4 , wherein an amino acid sequence of said BIM polypeptide is more than about 95% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         8 . The method of  claim 1 , wherein the cancer is breast cancer. 
     
     
         9 . The method of  claim 8 , wherein the breast cancer is an invasive breast carcinoma. 
     
     
         10 . The method of  claim 9 , wherein the invasive breast carcinoma is invasive ductal carcinoma. 
     
     
         11 . The method of  claim 1 , wherein the cancer is prostate cancer. 
     
     
         12 . The method of  claim 1 , wherein the cancer is ovarian cancer. 
     
     
         13 . The method of  claim 1 , wherein the cancer is pancreatic cancer. 
     
     
         14 . The method of  claim 1 , wherein the cancer is renal cancer. 
     
     
         15 . The method of  claim 1 , wherein the cancer is leukemia. 
     
     
         16 . The method of  claim 1 , wherein the cancer is Ph+ acute lymphocytic leukemia. 
     
     
         17 . A method of treating cancer in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle, wherein the cancer is colon cancer. 
     
     
         18 . The method of  claim 17 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         19 . The method of  claim 17 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         20 . The method of  claim 19 , wherein the peptidomimetic macrocycle is a BID polypeptide. 
     
     
         21 . The method of  claim 20 , wherein an amino acid sequence of said BID polypeptide is more than about 60% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         22 . The method of  claim 20 , wherein an amino acid sequence of said BID polypeptide is more than about 80% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         23 . The method of  claim 20 , wherein an amino acid sequence of said BID polypeptide is more than about 95% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         24 . A method of treating cancer in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle wherein said peptidomimetic macrocycle shows an EC 50  lower than 5 μM when tested in an in vitro cell viability assay against a cell line derived from said cancer. 
     
     
         25 . The method of  claim 24 , wherein the EC 50  is lower than 3 μM. 
     
     
         26 . The method of  claim 24 , wherein the cancer is selected from the group consisting of ovarian cancer, skin cancer, prostate cancer, renal cancer, breast cancer, pancreatic cancer, small-cell lung cancer, colon cancer, liver cancer, Multiple myeloma, Burkitt's lymphoma, acute lymphocytic leukemia (ALL) of T cell lineage or B cell lineage or mixed lineage, Chronic lymphocytic leukemia (CLL), Cutaneous T cell lymphoma (CTCL), Acute myelocytic leukemia (AML), Chronic Myelocytic leukemia, and follicular lymphoma. 
     
     
         27 . The method of  claim 26 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         28 . The method of  claim 26 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         29 . The method of  claim 26 , wherein the peptidomimetic macrocycle is a BIM polypeptide. 
     
     
         30 . The method of  claim 29 , wherein the BIM polypeptide is more than about 60% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         31 . The method of  claim 29 , wherein the BIM polypeptide is more than about 80% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         32 . The method of  claim 29 , wherein the BIM polypeptide is more than about 95% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         33 . The method of  claim 24 , wherein the cancer is selected from the group consisting of colon cancer, small-cell lung cancer, liver cancer, ovarian cancer, skin cancer, prostate cancer, renal cancer, breast cancer, pancreatic cancer, Multiple myeloma, Burkitt's lymphoma, acute lymphocytic leukemia (ALL) of T cell lineage or B cell lineage or mixed lineage, Chronic lymphocytic leukemia (CLL), Cutaneous T cell lymphoma (CTCL), Acute myelocytic leukemia (AML), Chronic Myelocytic leukemia and follicular lymphoma. 
     
     
         34 . The method of  claim 33 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         35 . The method of  claim 33 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         36 . The method of  claim 33 , wherein the peptidomimetic macrocycle is a BID polypeptide. 
     
     
         37 . The method of  claim 36 , wherein the BID polypeptide is more than about 60% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2 and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         38 . The method of  claim 36 , wherein the BID polypeptide is more than about 80% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         39 . The method of  claim 36 , wherein the BID polypeptide is more than about 95% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         40 . A method of treating a disorder in a human patient in need thereof comprising:
 a) preparing a peptidomimetic macrocycle by introducing a cross-link between two amino acid residues of a polypeptide;   b) testing the peptidomimetic macrocycle for the presence or absence of an immunogenic response; and   c) administering the peptidomimetic macrocycle to a patient if said immunogenic response does not cause a substantial side-effect.   
     
     
         41 . The method of  claim 40 , wherein said immunogenic response is evidenced as minimal antibody response in an in vivo assay in rodents. 
     
     
         42 . The method of  claim 40 , wherein the disorder is cancer. 
     
     
         43 . The method of  claim 40 , wherein the disorder is a metabolic disorder. 
     
     
         44 . The method of  claim 40 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         45 . The method of  claim 40 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         46 . A method of treating an immunoproliferative disorder in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle. 
     
     
         47 . The method of  claim 46 , wherein the peptidomimetic macrocycle comprises an α-helix. 
     
     
         48 . The method of  claim 46 , wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         49 . The method of  claim 46 , wherein the peptidomimetic macrocycle is a BID polypeptide. 
     
     
         50 . The method of  claim 49 , wherein the BID polypeptide is more than about 60% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         51 . The method of  claim 49 , wherein the BID polypeptide is more than about 80% identical to a sequence DlIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         52 . The method of  claim 49 , wherein the BID polypeptide is more than about 95% identical to a sequence DIIRNIARHLA*VGD*NleDRSI (SEQ ID NO: 2) and wherein * is a tethered amino acid and Nle is norleucine. 
     
     
         53 . The method of  claim 46 , wherein the peptidomimetic macrocycle is a BIM polypeptide. 
     
     
         54 . The method of  claim 53 , wherein the BIM polypeptide is more than about 60% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         55 . The method of  claim 53 , wherein the BIM polypeptide is more than about 80% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         56 . The method of  claim 53 , wherein the BIM polypeptide is more than about 95% identical to an amino acid sequence IWIAQELR*IGD*FNAYYARR (SEQ ID NO: 1) and wherein * is a tethered amino acid. 
     
     
         57 . The method of  claim 46 , wherein the peptidomimetic macrocycle reduces activated hPBL proliferation. 
     
     
         58 . The method of  claim 57 , wherein the peptidomimetic macrocycle reduces activated hPBL proliferation by more than 20% in an in vitro BrdU incorporation assay. 
     
     
         59 . The method of  claim 46 , wherein said immunoproliferative disease is a lymphoproliferative disorder. 
     
     
         60 . The method of  claim 1 ,  17 ,  24 ,  40  or  46  wherein an α-carbon atom in said peptidomimetic macrocycle is additionally substituted with independent substituents of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         61 . The method of  claim 60 , wherein an α-carbon atom to which the crosslinker is attached is additionally substituted with a substituent of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         62 . The method of  claim 60 , wherein an α-carbon atom to which the crosslinker is not attached is additionally substituted with a substituent of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         63 . The method of  claim 1 ,  17 ,  24 ,  40  or  46 , wherein two α-carbon atoms in said peptidomimetic macrocycle are additionally substituted with independent substituents of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         64 . The method of  claim 63 , wherein two α-carbon atoms to which the crosslinker is attached are additionally substituted with independent substituents of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         65 . The method of  claim 63 , wherein two α-carbon atoms to which the crosslinker is not attached are additionally substituted with independent substituents of formula R—, wherein R— is alkyl, alkenyl, alkynyl, arylalkyl, cycloalkylalkyl, heteroalkyl, or heterocycloalkyl, unsubstituted or substituted with halo-. 
     
     
         66 . The method of  claim 60  or  63 , wherein R— is alkyl. 
     
     
         67 . The method of  claim 60  or  63 , wherein R— is methyl. 
     
     
         68 . The method of  claim 60  or  63 , wherein the crosslinker connects two α-carbon atoms. 
     
     
         69 . The method of  claim 60  or  63 , wherein R— and any portion of the crosslinker taken together form a cyclic structure. 
     
     
         70 . The method of  claim 60  or  63 , wherein the crosslinker is formed of consecutive carbon-carbon bonds. 
     
     
         71 . The method of  claim 60  or  63 , wherein the crosslinker contains about 9 consecutive bonds. 
     
     
         72 . The method of  claim 60  or  63 , wherein the crosslinker contains about 12 consecutive bonds. 
     
     
         73 . The method of  claim 60  or  63 , wherein the crosslinker comprises at least about 6 carbon atoms. 
     
     
         74 . The method of  claim 60  or  63 , wherein the crosslinker comprises at least about 9 carbon atoms. 
     
     
         75 . The method of  claim 1 ,  17 ,  24 ,  40  or  46  wherein the peptidomimetic macrocycle is administered in conjunction with a standard method of care. 
     
     
         76 . The method of  claim 75 , wherein the standard method of care is chemotherapy. 
     
     
         77 . The method of  claim 75 , wherein the standard method of care is radiation therapy. 
     
     
         78 . The method of  claim 75 , wherein the standard method of care is surgery. 
     
     
         79 . The method of  claim 1 ,  17 ,  24 ,  40  or  46 , wherein the peptidomimetic macrocycle is cell permeable. 
     
     
         80 . The method of  claim 24 , wherein the assay is performed in the presence of 10% serum. 
     
     
         81 . The method of  claim 80 , wherein the serum is human serum. 
     
     
         82 . The method of  claim 24 , wherein the peptidomimetic macrocycle possesses an affinity of less than 10 μM for Mcl-1. 
     
     
         83 . The method of  claim 82 , wherein the peptidomimetic macrocycle antagonizes the interaction between Mcl-1 and a pro-apoptotic protein. 
     
     
         84 . The method of  claim 24 , wherein the cancer is resistant to a compound that possesses an affinity greater than 10 μM for Mcl-1. 
     
     
         85 . The method of  claim 84 , wherein the cancer is resistant to ABT-737 or an analog thereof. 
     
     
         86 . A method of treating ABT-737 resistant small cell lung cancer in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises a BH3 domain. 
     
     
         87 . A method of treating prostate cancer in a human patient in need thereof comprising administering to the patient a peptidomimetic macrocycle, wherein the peptidomimetic macrocycle comprises a BH3 domain.