FC Engineered Anti-TNFR Superfamily Member Antibodies Having Enhanced Atonistic Activity and Methods of Using Them

Abstract

The present invention relates to engineered anti-TNFR superfamily member antibodies having enhanced agonistic activity and methods of using them.

We claim: 
     
         1 . An isolated anti-tumor necrosis factor receptor (TNFR) superfamily member antibody, wherein the antibody comprises an E345R mutation, an E345R/E430G mutation or a E345R/E430G/S440Y mutation, residue numbering according to the EU Index, and has enhanced agonistic activity when compared to a parental antibody without the mutation. 
     
     
         2 . The antibody of  claim 1 , comprising the E345R mutation. 
     
     
         3 . The antibody of  claim 1 , comprising the E345R/E430G mutation. 
     
     
         4 . The antibody of  claim 1 , comprising the E345R/E430G/S440Y mutation. 
     
     
         5 . The antibody of  claim 1 , wherein the antibody has agonistic activity independent of antibody cross-linking. 
     
     
         6 . The antibody of  claim 5 , wherein the antibody is an IgG1, IgG2, IgG3 or IgG4 isotype. 
     
     
         7 . The antibody of  claim 6 , wherein the antibody comprises an amino acid sequence of SEQ ID NOs: 63, 64, 65, 66 or 67. 
     
     
         8 . The antibody of  claim 6 , further comprising a second mutation. 
     
     
         9 . The antibody of  claim 7 , wherein the second mutation is a L234A/L235A mutation on IgG1, a V234A/G237A/P238S/H268A/V309L/A330S/P331S mutation on IgG2, a F234A/L235A mutation on IgG4, a S228P/F234A/L235A mutation on IgG4, a N297A mutation on all Ig isotypes, a V234A/G237A mutation on IgG2, a K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M mutation on IgG1, a H268Q/V309L/A330S/P331S mutation on IgG2, a L234F/L235E/D265A mutation on IgG1, a L234A/L235A/G237A/P238S/H268A/A330S/P331S mutation on IgG1, a S228P/F234A/L235A/G237A/P238S mutation on IgG4, or a S228P/F234A/L235A/G236-deleted/G237A/P238S mutation on IgG4. 
     
     
         10 . The antibody of  claim 9 , wherein the second mutation is the V234A/G237A/P238S/H268A/V309L/A330S/P331S mutation on IgG2. 
     
     
         11 . The antibody of  claim 9 , wherein the second mutation is the L234A/L235A/G237A/P238S/H268A/A330S/P331S mutation on IgG1 
     
     
         12 . The antibody of  claim 9 , wherein the second mutation is the S228P/F234A/L235A mutation on IgG4. 
     
     
         13 . The antibody of  claim 1 , wherein the receptor of the TNFR family is OX40 (SEQ ID NO: 4), CD27 (SEQ ID NO: 8), CD40 (SEQ ID NO: 5), CD137 (SEQ ID NO: 10), or GITR (SEQ ID NO: 23). 
     
     
         14 . A pharmaceutical composition comprising the antibody of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         15 . A method of enhancing an agonistic activity of an anti-TNFR superfamily member antibody in a subject, comprising introducing an E345R mutation, an E345R/E430G mutation or an E345R/E430G/S440Y mutation into the antibody to generate an engineered antibody specifically binding the TNFR superfamily member, and administering the engineered antibody to the subject. 
     
     
         16 . The method of  claim 15 , wherein the antibody further comprises a second mutation. 
     
     
         17 . The method of  claim 16 , wherein the second mutation is a L234A/L235A mutation on IgG1, a V234A/G237A/P238S/H268A/V309L/A330S/P331S mutation on IgG2, a F234A/L235A mutation on IgG4, a S228P/F234A/L235A mutation on IgG4, a N297A mutation on all Ig isotypes, a V234A/G237A mutation on IgG2, a K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M mutation on IgG1, a H268Q/V309L/A330S/P331S mutation on IgG2, a L234F/L235E/D265A mutation on IgG1, a L234A/L235A/G237A/P238S/H268A/A330S/P331S mutation on IgG1, a S228P/F234A/L235A/G237A/P238S mutation on IgG4, or a S228P/F234A/L235A/G236-deleted/G237A/P238S mutation on IgG4. 
     
     
         18 . The method of  claim 17 , wherein the second mutation is the V234A/G237A/P238S/H268A/V309L/A330S/P331S mutation on IgG2. 
     
     
         19 . The method of  claim 17 , wherein the second mutation is the L234A/L235A/G237A/P238S/H268A/A330S/P331S mutation on IgG1 
     
     
         20 . The method of  claim 17 , wherein the second mutation is the S228P/F234A/L235A mutation on IgG4. 
     
     
         21 . The method of  claim 15 , wherein the subject has a cancer. 
     
     
         22 . The method of  claim 21 , wherein the cancer is a solid tumor. 
     
     
         23 . The method of  claim 22 , wherein the solid tumor is a melanoma, a lung cancer, a squamous non-small cell lung cancer (NSCLC), a non-squamous NSCLC, a colorectal cancer, a prostate cancer, a castration-resistant prostate cancer, a stomach cancer, an ovarian cancer, a gastric cancer, a liver cancer, a pancreatic cancer, a thyroid cancer, a squamous cell carcinoma of the head and neck, a carcinoma of the esophagus or gastrointestinal tract, a breast cancer, a fallopian tube cancer, a brain cancer, an urethral cancer, a genitourinary cancer, an endometriosis, a cervical cancer or a metastatic lesion of the cancer. 
     
     
         24 . The method of  claim 23 , wherein the TNFR superfamily member is OX40 (SEQ ID NO: 4), CD27 (SEQ ID NO: 8), CD40 (SEQ ID NO: 5), CD137 (SEQ ID NO: 10), or GITR (SEQ ID NO: 23). 
     
     
         25 . A method of treating a cancer in a subject, comprising administering to the subject an anti-TNFR superfamily member antibody comprising an E345R mutation, an E345R/E430G mutation or an E345R/E430G/S440Y mutation for a time sufficient to treat the cancer. 
     
     
         26 . The method of  claim 25 , wherein the antibody further comprises a second mutation. 
     
     
         27 . The method of  claim 26 , wherein the second mutation is a L234A/L235A mutation on IgG1, a V234A/G237A/P238S/H268A/V309L/A330S/P331S mutation on IgG2, a F234A/L235A mutation on IgG4, a S228P/F234A/L235A mutation on IgG4, a N297A mutation on all Ig isotypes, a V234A/G237A mutation on IgG2, a K214T/E233P/L234V/L235A/G236-deleted/A327G/P331A/D365E/L358M mutation on IgG1, a H268Q/V309L/A330S/P331S mutation on IgG2, a L234F/L235E/D265A mutation on IgG1, a L234A/L235A/G237A/P238S/H268A/A330S/P331S mutation on IgG1, a S228P/F234A/L235A/G237A/P238S mutation on IgG4, or a S228P/F234A/L235A/G236-deleted/G237A/P238S mutation on IgG4. 
     
     
         28 . The method of  claim 25 , wherein the cancer is a melanoma, a lung cancer, a squamous non-small cell lung cancer (NSCLC), a non-squamous NSCLC, a colorectal cancer, a prostate cancer, a castration-resistant prostate cancer, a stomach cancer, an ovarian cancer, a gastric cancer, a liver cancer, a pancreatic cancer, a thyroid cancer, a squamous cell carcinoma of the head and neck, a carcinoma of the esophagus or gastrointestinal tract, a breast cancer, a fallopian tube cancer, a brain cancer, an urethral cancer, a genitourinary cancer, an endometriosis, a cervical cancer or a metastatic lesion of the cancer. 
     
     
         29 . The method of  claim 28 , wherein the receptor of the TNFR family is OX40 (SEQ ID NO: 4), CD27 (SEQ ID NO: 8), CD40 (SEQ ID NO: 5), CD137 (SEQ ID NO: 10), or GITR (SEQ ID NO: 23).