Conjugates of membrane translocating agents and pharmaceutically active agents

Abstract

Membrane translocation peptides, compositions comprising them, chimeric molecules comprising them, and methods of using them to achieve transmembrane transport of various agents.

We claim:  
     
         1 . A composition comprising a translocating peptide, said translocating peptide selected from the group consisting of a transport peptide, an extended peptide comprising said transport peptide, and a transport-active fragment of at least 4 amino acids of said transport peptide, wherein said transport peptide is selected from the group consisting of an L-peptide, a d-peptide, and a retroinverted peptide, and 
 wherein said L-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS: 2-13, and 15-24,    wherein said d-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS. 102-124 corresponding to the d-forms of L-peptides of SEQ ID NOS. 2-24, and    wherein the retroinverted peptide has an amino acid sequence selected from the group consisting of a peptide of SEQ ID NOS. 202-224, corresponding to retroinverted forms of L-peptides of SEQ ID NOS: 2-24.    
     
     
         2 . A composition of  claim 1 , 
 wherein said L-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS: 2-4, 16, 23 and 24.    wherein said d-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS. 102-104, 116, 123 and 124 corresponding to the d-forms of L-peptides of SEQ ID NOS. 2-4,16, 23 and 24,    wherein the retroinverted peptide has an amino acid sequence selected from the group consisting of a peptide of SEQ ID NOS. 202-204, 216, 223 and 224, corresponding to retroinverted forms of an L-peptides of SEQ ID NOS: 2-4, 16, 23 and 24.    
     
     
         3 . A composition of  claim 1 , wherein said transport peptide is partially or completely cyclic.  
     
     
         4 . A composition of  claim 3  wherein any fragment of said transport peptide is also partially or completely cyclic.  
     
     
         5 . A composition of  claim 4  where in the 
 wherein said L-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS: 5-13;  
 wherein said d-peptide has an amino acid sequence selected from the group consisting of SEQ ID NOS. 105-113 corresponding to the d-forms of L-peptides of SEQ ID NOS. 5-13,  
 wherein the retroinverted peptide has an amino acid sequence selected from the group consisting of a peptide of SEQ ID NOS. 205-213, corresponding to retroinverted forms of L-peptides of SEQ ID NOS: 5-13.  
 
     
     
         6 . A composition of  claim 1 , 
 wherein said translocating peptide is an extended peptide of a transport peptide.    
     
     
         7 . A composition of  claim 6  wherein the extended peptide is not more than 100 amino acids in length.  
     
     
         8 . A composition of  claim 7  wherein the extended peptide is not more than 50 amino acids in length.  
     
     
         9 . A composition of  claim 1  wherein the translocating peptide is a transport peptide.  
     
     
         10 . A composition of  claim 1  where in the transport-active fragment is at least 6 amino acids of a transport peptide.  
     
     
         11 . A composition of  claim 1  where in the transport-active fragment is at least 8 amino acids of a transport peptide.  
     
     
         11 A. A composition of  claim 1  wherein the translocating peptide is selected from the group consisting of Elan 094, Elan178, Elan207, and Elan208.  
     
     
         12 . A composition of  claim 1  wherein the carboxyl end group of the translocating peptide has been modified to create an amide group.  
     
     
         13 . A composition comprising a translocating peptide, said translocating peptide selected from the group consisting of a transport peptide, an extended peptide comprising said transport peptide, and a transport-active fragment of at least 4 amino acids of said transport peptide, said transport peptide being an L-peptide that has an amino acid sequence SEQ ID NO: 14 blocked at its carboxyl end with an amide group and wherein any of said fragments is also blocked at its carboxyl end with an amide group.  
     
     
         14 . The composition of claims  1  or  13 , further comprising an active agent, wherein the translocating peptide is complexed to an active agent to form a translocating peptide-active agent complex.  
     
     
         15 . The composition of claims  1  or  13 , further comprising an active particle, wherein the translocating peptide is complexed to the active particle to form a translocating peptide-active particle complex.  
     
     
         16 . A method for enhancing movement of an active agent across a lipid membrane, comprising using a translocating peptide-active agent complex, wherein the translocating peptide enhances movement of the active agent across the lipid membrane.  
     
     
         17 . A method for enhancing movement of an active particle across a lipid membrane, comprising using a translocating peptide-active particle complex, wherein the translocating peptide enhances movement of the active particle across the lipid membrane.  
     
     
         18 . A method for identifying a compound having enhanced ability to transport an active agent across a lipid membrane, wherein the compound competes with the translocating peptide for transport of an fMLP across a membrane selected from the group consisting of a cell membrane, an intracellular membrane, the apical and basal membranes of an epithelial cell layer.  
     
     
         19 . The method of  claim 18 , wherein the epithelial cell layer is a polarized epithelial cell layer.  
     
     
         20 . A method for treating a pathological disorder in an animal, comprising orally administering to the animal in need of such treatment a complex selected from the group consisting of a translocating peptide-active agent complex and a translocating peptide-active particle complex, wherein an amount of the active agent effective to treat the pathological disorder is moved across the gastrointestinal epithelium of the animal into the circulation.  
     
     
         21 . A chimeric polypeptide comprising (A) a translocating peptide of claims  1  or  13 , (B) a translocatable peptide, and (C) an amino acid linker sequence that directly links the translocating peptide to the translocatable peptide, wherein said translocatable peptide is between 3 and 200 amino acids, and wherein said amino acid linker sequence is between 1 and 20 amino acids.  
     
     
         22 . A chimeric peptide of  claim 21  wherein said translocatable peptide is between 3 and 30 amino acids.  
     
     
         23 . A chimeric peptide of  claim 21  wherein the translocatable peptide is an opioid peptide.  
     
     
         24 . A chimeric peptide of  claim 21  wherein said linker sequence is not more than 7 amino acids.  
     
     
         25 . A chimeric peptide of  claim 24  wherein said linker sequence is not more than 3 amino acids.  
     
     
         26 . A chimeric peptide of  claim 25  wherein said linker sequence is 1 amino acids.  
     
     
         27 . A chimeric peptide of  claim 26  wherein said at least 50% of the amino acids in the linker sequence are lysines.  
     
     
         28 . A chimeric peptide of  claim 26  wherein said at least 80% of the amino acids in the linker sequence are lysines.  
     
     
         29 . A chimeric peptide of  claim 26  wherein all of the amino acids in the linker sequence are lysines.  
     
     
         30 . A method of delivering a chimeric peptide to the blood, said method comprising orally administering a chimeric peptide of  claim 21 .  
     
     
         31 . A nucleic acid molecule coding for a translocating peptide of  claim 1 .  
     
     
         32 . A nucleic acid molecule of  claim 31  wherein the translocating peptide is an L-form peptide.  
     
     
         33 . A nucleic acid molecule coding for a chimeric protein of  claim 21 .  
     
     
         34 . A nucleic acid molecule of  claim 33  wherein the chimerica peptide is an L-form peptide.  
     
     
         35 . A chimeric constructs comprising (A) a translocating peptide of claims  1  or  13 , (B) a translocatable peptide, and (C) an non-amino acid linker that directly links the translocating peptide to the translocatable peptide, wherein said translocatable peptide is between 3 and 200 amino acids.  
     
     
         36 . A method of delivering a chimeric construct to a site within a person, said method comprising administering a chimeric construct of  claim 35 , said site being selected from the group consisting of a tissue, a fluid, a cell, and a sub-cellular compartment.