Conjugation of pharmaceutically active agents with transthyretin ligands through adjustable linkers to increase serum half-life

Abstract

A delivery system for active agents, and methods of making and using the systems, are provided. The delivery systems have (i) a ligand that is selective for an endogeneous plasma protein in the serum of a subject; and, (ii) a linker configured for operatively attaching the ligand covalently to an active agent to increase the half-life of the active agent in the serum.

1 . A delivery system for an active agent, comprising:
 a ligand with (i) a high selectivity for a plasma protein endogeneous to the subject, the molecular weight of the plasma protein ranging from about 30 kDa to about 80 kDa; (ii) a high binding affinity, Kd, of at least 10 −6  M for the plasma protein; and, (iii) a molecular weight ranging from about 200 Da to about 2000 Da;   and,   a linker that ranges in length from about 10 angstroms to about 50 angstroms, or from 8 atoms to 50 atoms;   wherein, the active agent has a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an imaging agent, and combinations thereof.   
     
     
         2 . (canceled) 
     
     
         3 . (canceled) 
     
     
         4 . The delivery system of  claim 1 , wherein the plasma protein is TTR. 
     
     
         5 . A delivery system for an active agent, comprising:
 a ligand that is selective for transthyretin in the serum of a subject; and,   a linker configured for operatively attaching the ligand covalently to an active agent, wherein the linker ranges in length from 14 angstroms to 30 angstroms, or from 10 atoms to 22 atoms;   wherein,
 the ligand has the following structure of Compound (I) 
   
       
         
           
           
               
               
           
         
         
           where X a , X b  and X c  are independently selected from C(R 4 )(R 5 ), O, N—R 5  or S; where R 4  and R 5  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
           a ring is a 4 to 12-membered ring, in certain embodiments the 4 to 12-membered ring is an aromatic or heteroaromatic ring; 
           each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, sulfonamide, sulfonyl fluoride, thioester and cyano; 
           c is an integer ranging from 0 to 5; and, 
           B ring is a hetercyclic ring selected from the following (h1-h30): 
         
       
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           where R 11 -R 16  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; R 17  is selected from a hydroxyl, alkyl, amino, and alkyl amino; and at least one of R 11 -R 16  is the linking group to X c ; 
           or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, amide, acetal, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
         
       
     
     
         6 . The delivery system of  claim 5 , wherein the ligand has the following structure of Compound (II), comprising: 
       
         
           
           
               
               
           
         
         where,
 n is an integer ranging from 0 to 8; 
 R 1 , R 2  and R 3  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, sulfonamide, sulfonyl fluoride, thioester and cyano; 
 X a  is C(R 4 )(R 5 ), O, N—R 5  or S; where R 4  and R 5  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 A is a 5 to 12-membered ring, in certain embodiments the 5 to 12-membered ring is an aromatic or heteroaromatic ring; 
 each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, sulfonamide, sulfonyl fluoride, thioester and cyano; and, 
 c is a number from zero to 5; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, amide, acetal, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         7 . The delivery system of  claim 5 , wherein the ligand has the structure of Compound (Ill), comprising: 
       
         
           
           
               
               
           
         
         where,
 n is an integer ranging from 0 to 7; 
 Z is carbon and/or up to three of the five Z may be nitrogen; 
 R1, R2 and R3 are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 Xa is C(R4)(R5), O, N—R5 or S; where R4 and R5 are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 each Y is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, sulfonamide, sulfonyl fluoride, thioester and cyano; and 
 c is an integer ranging from 0 to 5; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, amide, acetal, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         8 . (canceled) 
     
     
         9 . The delivery system of  claim 6 ; wherein, the ligand has the structure of Compound (IV), comprising: 
       
         
           
           
               
               
           
         
         where,
 n is an integer ranging from 1 to 4; 
 R 1  is a short chain alkyl having 1 to 4 carbon atoms; 
 R 2  is hydrogen; 
 R 3  is a short chain alkyl having 1 to 4 carbon atoms; 
 X a  is C(R 4 )(R 5 ), O, N—R 5  or S; where R 4  and R 5  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 each Y is independently selected from hydrogen, halogen, acyl, substituted acyl, carboxyl, heterocyclic group, alkoxycarbonyl sulfonamide, sulfonyl fluoride, thioester and substituted alkoxycarbonyl; and 
 c is 2; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         10 . The delivery system of  claim 9 , wherein:
 R1 is methyl and R3 is methyl;   Xa is O, and,   Y is fluoro or carboxyl.   
     
     
         11 . The delivery system of  claim 7 , wherein the compound has structure of Compound (V), comprising: 
       
         
           
           
               
               
           
         
         where,
 n is 1 to 8; 
 R 1 , R 2  and R 3  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halo, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 X a  is C(R 4 )(R 5 ), O, N—R 5  or S; where R 4  and R 5  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 R a  is CHO, COOH, COOCH 3 , COOR 6 , CONR 7 R 8 , tetrazolyl, CONHOH, B(OH) 2 , CONHSO 2 Ar, CONHCH(R 9 )COOH, CF 3 , hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic group, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; 
 R b  is CHO, COOH, COOCH 3 , COOR 6 , CONR 7 R 8 , tetrazolyl, CONHOH, B(OH) 2 , CONHSO 2 Ar, CONHCH(R 9 )COOH, CF 3 , hydrogen, halogen, alkyl, substituted alkyl, acyl, substituted acyl, carboxyl, heterocyclic group, sulfonamide, sulfonyl fluoride, thioester, alkoxycarbonyl or substituted alkoxycarbonyl; 
 R 6  is alkyl, haloalkyl, cycloalkyl, or heterocyclyl; 
 R 7  and R 8  are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, or heteroaryl; and, 
 R 9  is the side chain of a naturally occurring α-amino carboxylic acid; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, amide, acetal, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         12 . The delivery system of  claim 11 , wherein R b  is selected from bromo, chloro and fluoro. 
     
     
         13 . The delivery system of  claim 6 , wherein the ligand has the structure of Compound (VI), comprising: 
       
         
           
           
               
               
           
         
         where,
 n is 3; 
 R 1  is a short chain alkyl having 1 to 4 carbon atoms; 
 R 2  is hydrogen; 
 R 3  is a short chain alkyl having 1 to 4 carbon atoms; 
 X a  is C(R 4 )(R 5 ), O, N—R 5  or S; where R 4  and R 5  are independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, alkoxy, aryloxy, hydroxyl, heterocyclic group, halogen, nitro, acyl, substituted acyl, carboxyl, alkoxycarbonyl, substituted alkoxycarbonyl, aminoacyl, substituted aminoacyl, amino, substituted amino, acylamino, substituted acylamino, and cyano; 
 R a  is CHO, COOH, COOCH 3 , COOR 6 , CONR 7 R 8 , tetrazolyl, CONHOH, B(OH) 2 , CONHSO 2 Ar, CONHCH(R 9 )COOH, hydrogen, an acyl, substituted acyl, carboxyl, alkoxycarbonyl, heterocyclic group, sulfonamide, sulfonyl fluoride, thioester, or substituted alkoxycarbonyl; 
 R b  is CHO, COOH, COOCH 3 , COOR 6 , CONR 7 R 8 , tetrazolyl, CONHOH, B(OH) 2 , CONHSO 2 Ar, CONHCH(R 9 )COOH, a halogen or heterocyclic group; 
 R 6  is alkyl, haloalkyl, cycloalkyl, or heterocyclyl; 
 R 7  and R 8  are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, or heteroaryl; and 
 R 9  is the side chain of a naturally occurring α-amino carboxylic acid; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         14 . The delivery system of  claim 6 , wherein the ligand has the structure of Compound (VIIc): 
       
         
           
           
               
               
           
         
         or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
       
     
     
         15 . The delivery system of  claim 5 , wherein the ligand has the structure of Compound (VIIa), comprising: 
       
         
           
           
               
               
           
         
         where,
 R a  is OH, CHO, COOH, CONH 2 , CONH(OH), COOR 6 , CONHR 6 ; 
 R 6  is straight of branched alkyl of 1-3 carbon atoms; 
 or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         16 . The delivery system of  claim 5 , wherein the ligand has the structure of Compound (VIIb), comprising: 
       
         
           
           
               
               
           
         
         where,
 R a  is COOH, CONH 2 , CONH(OH), COOR 6 , CONHR 6 ; 
 R 6  is straight of branched alkyl of 1-3 carbon atoms; 
 
         or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
       
     
     
         17 . The delivery system of  claim 5 , wherein the ligand is selected from the group consisting of: 
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         and, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide,
 and, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof. 
 
       
     
     
         18 . The delivery system of  claim 6 , wherein the ligand has the structure of Compound (VIIIc): 
       
         
           
           
               
               
           
         
         or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof; and, 
         the linker is attached to the ligand ortho at C15 to the carboxyl group at C14. 
       
     
     
         19 . The delivery system of  claim 6 , wherein the ligand has the structure of Compound (VIIIc): 
       
         
           
           
               
               
           
         
         or, a pharmaceutically acceptable salt, ester, enol ether, enol ester, acetal, amide, ketal, orthoester, hemiacetal, hemiketal, hydrate, solvate or prodrug thereof; and, 
         the linker is attached to the ligand meta at C16 to the carboxy carbon at C14. 
       
     
     
         20 . A method of increasing the in vivo half-life of an active agent, the method comprising covalently attaching the delivery system of  claim 5  to an active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         21 . A method of increasing the in vivo half-life of an active agent, the method comprising covalently attaching the delivery system of  claim 6  to an active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         22 . A method of increasing the in vivo half-life of an active agent in the blood serum of a subject, the method comprising covalently attaching the delivery system of  claim 7  to an active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         23 . A method of increasing the in vivo half-life of an active agent in the blood serum of a subject, the method comprising covalently attaching the delivery system of  claim 15  to a drug, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         24 . A method of increasing the in vivo half-life of an active agent in the blood serum of a subject, the method comprising covalently attaching the delivery system of  claim 18  to an active agent; wherein, the active agent comprises a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         25 . A method of increasing the in vivo half-life of an active agent in the blood serum of a subject, the method comprising covalently attaching the delivery system of  claim 19  to an active agent; wherein, the active agent comprises a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof. 
     
     
         26 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 5  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         27 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 6  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         28 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 7  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         29 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 15  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an oligosaccharide, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         30 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 18  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         31 . A method of administering an active agent to a subject, the method comprising:
 covalently attaching the delivery system of  claim 19  to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an imaging agent, and combinations thereof; and,   administering the conjugated active agent to the subject.   
     
     
         32 . A method of reducing the immunogenicity of an active agent in a subject, the method comprising:
 obtaining a delivery system having
 a ligand with (i) a high selectivity for a plasma protein endogeneous to the subject, the molecular weight of the plasma protein ranging from about 30 kDa to about 80 kDa; (ii) a high binding affinity, Kd, of at least 10 −6  M for the plasma protein; and, (iii) a molecular weight ranging from about 200 Da to about 2000 Da; 
 and, 
 a linker the linker ranges in length from about 10 angstroms to about 50 angstroms, or from 10 atoms to 50 atoms; 
   covalently attaching the delivery system to an active agent to create a conjugated active agent, the active agent comprising a structure selected from the group consisting of a peptide, an oligopeptide, a polypeptide, a protein, an antibody, an oligonucleotide, a polynucleotide, a virus-like particle, a small molecule, an imaging agent, and combinations thereof;   and,   administering the conjugated active agent to the subject;   wherein, the plasma protein shields the active agent from antibody generation in the subject after the administering.   
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . The method of  claim 32 , wherein the plasma protein is TTR. 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . (canceled) 
     
     
         40 . (canceled) 
     
     
         41 . (canceled)