US2015218280A1PendingUtilityA1

CD20 scFv-ELPs METHODS AND THERAPEUTICS

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Assignee: UNIV SOUTHERN CALIFORNIAPriority: Aug 10, 2012Filed: Aug 8, 2013Published: Aug 6, 2015
Est. expiryAug 10, 2032(~6 yrs left)· nominal 20-yr term from priority
Inventors:Alan L. EpsteinJohn Andrew MackayPeisheng HuSuhaas Aluri
C07K 2317/73A61K 48/005A61K 47/6435A61K 9/51C07K 2319/735A61K 2039/505A61K 47/6849A61K 47/6929C07K 2317/622C07K 2319/74A61K 9/1075C07K 16/2887C07K 2319/33C07K 14/78A61K 38/00A61K 47/48561
49
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Claims

Abstract

Disclosed herein are recombinant polypeptides comprising an elastin-like peptide (ELP) and a scFv, or a biological equivalent of the scFv. Also disclosed are compositions containing scFv-ELP polypeptides and methods of use.

Claims

exact text as granted — not AI-modified
1 . A recombinant polypeptide comprising an elastin-like peptide (ELP) fused to a scFv, or a biological equivalent of the scFv. 
     
     
         2 . The recombinant polypeptide of  claim 1 , wherein the scFv is fused to the N-terminus of ELP. 
     
     
         3 . The polypeptide of  claim 1  or  2 , wherein the scFv is the single chain variable region of the anti-CD20 antibody. 
     
     
         4 . The polypeptide of  claim 2 , wherein the sequence of the polypeptide corresponds to a sequence selected from the group consisting of: SEQ ID NOS: 3, 4, and 5 or a biological equivalent thereof. 
     
     
         5 . The polypeptide of  claim 1 , wherein the ELP comprises the primary sequence of SEQ ID NO. 6, wherein X is any amino acid and n is an integer from 1 to about 500 (SEQ ID NO: 14). 
     
     
         6 . The polypeptide of  claim 1 , wherein the ELP comprises the primary sequence of SEQ ID NO: 7, wherein n is an integer from 1 to about 500. 
     
     
         7 . The polypeptide of  claim 1 , wherein the ELP consists essentially of the primary sequence of SEQ ID NO: 6, wherein X is any amino acid (Repeat sequences of SEQ NO: 6 wherein n is an integer from 1 to about 500 is disclosed as SEQ ID NO:
 14) or SEQ ID NO: 7, wherein n is an integer from 1 to about 500.   
     
     
         8 . The polypeptide of  claim 1  further comprising a detectable label. 
     
     
         9 . The polypeptide of  claim 3 , wherein the scFv comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2 or a biological equivalent thereof. 
     
     
         10 . The polypeptide of  claim 1  linked to a therapeutic agent. 
     
     
         11 . The polypeptide of  claim 10 , wherein the therapeutic agent is an anti-cancer drug. 
     
     
         12 . An isolated polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         13 . A vector and/or host cell comprising the polynucleotide of  claim 12 . 
     
     
         14 . A composition comprising at least two polypeptides of  claim 1 . 
     
     
         15 . The composition of  claim 14 , wherein the at least two polypeptides are organized into a cylindrical particle. 
     
     
         16 . The composition of  claim 15 , wherein the cylindrical particle has a core. 
     
     
         17 . The composition of  claim 16 , wherein the core of the cylindrical particle comprises an scFv is fused to a N-terminus of an ELP. 
     
     
         18 . The composition of  claim 14 , wherein the at least two polypeptides are organized into a spherical particle or a nanoworm. 
     
     
         19 . The composition of  claim 18 , wherein the particle has a core. 
     
     
         20 . The composition of  claim 19 , wherein the core of the particle comprises an scFv is fused to a N-terminus of an ELP. 
     
     
         21 . A composition comprising a carrier and the polypeptide of  claim 1 . 
     
     
         22 . A method for preparing a therapeutic polypeptide, comprising expressing the polynucleotide of  claim 12  in a suitable expression system. 
     
     
         23 . The method of  claim 22 , further comprising separating or purifying the polypeptide from the expression system. 
     
     
         24 . The method of  claim 23 , further comprising denaturing and refolding the polypeptide. 
     
     
         25 . The method of  claim 24 , wherein the denaturing and refolding is performed at least twice. 
     
     
         26 . A method for inducing apoptosis of a CD20+ cell comprising contacting the cell with an effective amount of the polypeptide of  claim 3 . 
     
     
         27 . The method of  claim 26 , wherein the contacting is to a cell in vitro or in vivo. 
     
     
         28 . The method of  claim 26 , wherein the cell is a malignant B-cell. 
     
     
         29 . A method for treating a CD20-related disease or disorder, comprising administering to a patient in need of such treatment the polypeptide of  claim 3 . 
     
     
         30 . The method of  claim 29 , wherein the CD20-related disease or disorder is cancer or an autoimmune disease. 
     
     
         31 . A kit for treating a CD20-related disease or disorder or inducing apoptosis of a CD20+ cell, comprising the polypeptide of  claim 3 . 
     
     
         32 . A method for targeting a scFv-ELP to a cell expressing CD20 comprising contacting the cell with an effective amount of the polypeptide of  claim 1 , wherein the scFv component of the scFv-ELP binds to the CD20 receptor on the cell. 
     
     
         33 . The method of  claim 32  wherein the contacting is to a cell in vitro or in vivo. 
     
     
         34 . The method of  claim 33  wherein the scFv-ELP is linked to a therapeutic agent.

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