Antibody drug conjugate for anti-inflammatory applications

Abstract

Antibody drug conjugates (ADCs) comprising an antibody conjugated to an anti-inflammatory therapeutic agent via a phosphate-based linker with tunable extracellular and intracellular stability are described.

1 : A composition comprising a compound having formula (I) 
       
         
           
           
               
               
           
         
         wherein V is selected from O and S; 
         W is selected from O, N, and C H 2; 
         X is a tuning element selected from a covalent bond; a carbon atom; a heteroatom; 
       
       an optionally substituted group selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, heteroaryl, and heterocyclic; nucleoside, protease sensitive group, cathepsin B sensitive group, or glycosidase sensitive group;
 Y is selected from a covalent bond or a bivalent, straight or branched, saturated or unsaturated, optionally substituted C 1-30  hydrocarbon chain wherein one or more methylene units of Y are optionally and independently replaced by —O—, —S—, —N(R)—, —C(O)—, C(O)O—, OC(O)—, —N(R)C(O)—, —C(O)N(R)—, —S(O)—, —S(O) 2 —, —N(R)SO 2 —, SO 2 N(R)—, a heterocyclic group, an aryl group, or a heteroaryl group; 
 T is an NR, CR 2 , O, or S; 
 D is an anti-inflammatory agent; 
 antibody is a chimeric, humanized, or human antibody having a light chain and a heavy chain wherein the antibody binds the human CD74 protein or human CD163 protein; 
 Z is a linkage formed between (i) a reactive functional group selected from the group consisting of N-hydroxysuccinimide, para-nitrophenyl carbonate, para-nitrophenyl carbamate, pentafluorophenyl, haloacetamide, maleimide, hydroxylamine, strained cycloalkyne, and heterocycloalkyne, alkyne, diene, azadiene, and heterocyclic azadience, wherein halo is iodine (I), bromine (Br), fluorine (F), or chlorine (Cl) and hetero is N, O, or S, and (ii) the side chain of an amino acid in the heavy chain or light chain of the antibody; 
 each occurrence of R is independently hydrogen, a suitable protecting group, an acyl moiety, arylalkyl moiety, aliphatic moiety, aryl moiety, heteroaryl moiety, or heteroaliphatic moiety; 
 n is 1, 2, 3, or 4; 
 m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 
 and a pharmaceutically acceptable carrier. 
 
     
     
         2 : The compound of  claim 1 , wherein the anti-inflammatory agent comprises a glucocorticoid receptor agonist. 
     
     
         3 : The compound of  claim 1 , wherein the anti-inflammatory agent comprises Cortisol, cortisone acetate, beclometasone, prednisone, prednisolone, methylprednisolone, betamethasone, trimcinolone, budesonide, dexamethasone, fluticasone, or mometasone. 
     
     
         4 : The compound of  claim 1 , wherein the antibody comprises a substitution of an amino acid in the heavy chain or light chain of the antibody with para-azidophenylalanine (pAzF) and the reactive functional group comprises a strained cycloalkyne. 
     
     
         5 - 6 . (canceled) 
     
     
         7 : The composition of  claim 1 , wherein the antibody comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO:69, 70, 71, and 72 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:73. 
     
     
         8 - 12 . (canceled) 
     
     
         13 : The composition of  claim 1 , wherein the antibody comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO:74, 75, 76, and 77 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:78. 
     
     
         14 - 16 . (canceled) 
     
     
         17 : A method for treating an inflammatory disease or disorder by providing to a subject having the disease or disorder a composition comprising a compound having formula (I) 
       
         
           
           
               
               
           
         
         wherein V is selected from O and S; 
         W is selected from O, N, and C H 2; 
         X is a tuning element selected from a covalent bond; a carbon atom; a heteroatom; 
       
       an optionally substituted group selected from the group consisting of acyl, aliphatic, heteroaliphatic, aryl, heteroaryl, and heterocyclic; nucleoside, protease sensitive group, cathepsin B sensitive group, or glycosidase sensitive group;
 Y is selected from a covalent bond or a bivalent, straight or branched, saturated or unsaturated, optionally substituted C 1-30  hydrocarbon chain wherein one or more methylene units of Y are optionally and independently replaced by —O—, —S—, —N(R)—, —C(O)—, C(O)O—, OC(O)—, —N(R)C(O)—, —C(O)N(R)—, —S(O)—, —S(O) 2 —, —N(R)SO 2 —, SO 2 N(R)—, a heterocyclic group, an aryl group, or a heteroaryl group; 
 T is an NR, CR 2 , O, or S; 
 D is an anti-inflammatory agent; 
 antibody is a chimeric, humanized, or human antibody having a light chain and a heavy chain wherein the antibody binds the human CD74 protein or human CD163 protein; 
 Z is a linkage formed between (i) a reactive functional group selected from the group consisting of N-hydroxysuccinimide, para-nitrophenyl carbonate, para-nitrophenyl carbamate, pentafluorophenyl, haloacetamide, maleimide, hydroxylamine, strained cycloalkyne, and heterocycloalkyne, alkyne, diene, azadiene, and heterocyclic azadience, wherein halo is iodine (I), bromine (Br), fluorine (F), or chlorine (Cl) and hetero is N, O, or S, and (ii) the side chain of an amino acid in the heavy chain or light chain of the antibody; 
 each occurrence of R is independently hydrogen, a suitable protecting group, an acyl moiety, arylalkyl moiety, aliphatic moiety, aryl moiety, heteroaryl moiety, or heteroaliphatic moiety; 
 n is 1, 2, 3, or 4; 
 m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; 
 and a pharmaceutically acceptable carrier to treat the inflammatory disease or disorder. 
 
     
     
         18 : The method of  claim 17 , wherein the anti-inflammatory agent comprises a glucocorticoid receptor agonist. 
     
     
         19 : The method of  claim 17 , wherein the anti-inflammatory agent comprises Cortisol, cortisone acetate, beclometasone, prednisone, prednisolone, methylprednisolone, betamethasone, trimcinolone, budesonide, dexamethasone, fluticasone, or mometasone. 
     
     
         20 : The method of  claim 17 , wherein the antibody comprises a substitution of an amino acid in the heavy chain or light chain of the antibody with pAzF and the reactive functional group comprises a strained cycloalkyne. 
     
     
         21 : The method of  claim 17 , wherein the inflammatory disease or disorder comprises Alzheimer's disease, ankylosing spondylitis arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis), asthma, atherosclerosis, Crohn's disease, colitis, dermatitis, diverticulitis, fibromyalgia, hepatitis, irritable bowel syndrome (IBS), systemic lupus erythematous (SLE), nephritis, Parkinson's disease, or ulcerative colitis. 
     
     
         22 - 23 . (canceled) 
     
     
         24 : The method of  claim 17 , wherein the antibody comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO: 69, 70, 71, and 72 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:73. 
     
     
         25 - 29 . (canceled) 
     
     
         30 : The method of  claim 17 , wherein the antibody comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO:74, 75, 76, and 77 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:78. 
     
     
         31 - 50 . (canceled) 
     
     
         51 : An antibody drug conjugate comprising
 (a) an antibody that binds a CD74 protein, wherein the antibody comprises a para-azidophenylalanine (pAzF) conjugated to a molecule selected from the group of molecules consisting of 1-4, 2-7, 3-4, 4-3, 5-3, 6-2, 7-1, 8-5, 9-1, 10-1, 11-5, 21-1, 13-1, 14-5, 15-1, 16-5, and 17-2;   (b) an antibody that binds a CD163 protein, wherein the antibody comprises a para-azidophenylalanine (pAzF) conjugated to a molecule selected from the group of molecules consisting of 1-4, 2-7, 3-4, 4-3, 5-3, 6-2, 7-1, 8-5, 9-1, 10-1, 11-5, 21-1, 13-1, 14-5, 15-5, 16-5, and 17-2: or   (c) antibody drug conjugate selected from the group consisting of ADC 12-1, ADC 12-2, ADC 12-3, ADC 12-4, ADC 12-5, ADC 12-6, ADC 12-7, ADC 12-8, ADC 12-9, ADC 12-10, ADC 12-13, ADC 12-14, and ADC 12-15.   
     
     
         52 - 53 . (canceled) 
     
     
         54 : The antibody drug conjugate of  claim 51 , wherein the antibody that binds CD74 protein comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO:69, 70, 71, and 72 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:73. 
     
     
         55 - 59 . (canceled) 
     
     
         60 : The antibody drug conjugate of  claim 51 , wherein the antibody that binds CD74 protein comprises a heavy chain (HC) comprising an amino acid sequence selected from SEQ ID NO:74, 75, 76, and 77 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:78. 
     
     
         61 . (canceled) 
     
     
         62 : The antibody drug conjugate of  claim 51 , wherein the antibody that binds CD163 protein comprises light chain CDR sequences CDR1 (ASQSVSSDV; SEQ ID NO:60), CDR2 (YAS), and CDR3 (QDYTSPRT; SEQ ID NO:61) and heavy chain complementarity-determining region (CDR) sequences CDR1 (GYSITSDY; SEQ ID NO:62), CDR2 (YSG), and CDR3 (CVSGTYYFDYWG; SEQ ID NO:63). 
     
     
         63 : The antibody drug conjugate of  claim 51 , wherein the antibody that binds CD163 protein comprises light chain CDR sequences CDR1 (ASQSVSHDV; SEQ ID NO:54), CDR2 (YTS), and CDR3 (QDYSSPRT; SEQ ID NO:65) and heavy chain CDR sequences CDR1 (GYSITSDY; SEQ ID NO:62), CDR2 (YSG), and CDR3 (CVSGTYYFDYWG; SEQ ID NO:63). 
     
     
         64 - 65 . (canceled)