US2014315817A1PendingUtilityA1

Variant serum albumin with improved half-life and other properties

47
Assignee: ELEVEN BIOTHERAPEUTICS INCPriority: Nov 18, 2011Filed: Nov 18, 2012Published: Oct 23, 2014
Est. expiryNov 18, 2031(~5.3 yrs left)· nominal 20-yr term from priority
Inventors:Michael March SchmidtEric FurfineAmy Jada AndreucciThomas M. Barnes
C07K 16/18C07K 14/55C07K 14/755C07K 14/765A61K 38/385C12N 9/6437C07K 2319/31
47
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention provides methods and materials for making and using variant serum albumin amino acid sequences which exhibit improved properties compared to wild type serum albumin sequences. The invention further provides methods and materials for making and using fusion proteins in which the variant serum albumin amino acid sequences are fused to a therapeutic or diagnostic agent, such as a therapeutic protein, or a functional fragment or variant thereof that maintains activity, and exhibits improved properties.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . An isolated, recombinant protein comprising a variant serum albumin polypeptide (VSA) sequence that is a variant of domain III of a naturally-occurring serum albumin sequence, wherein the variant comprises a mutation at one or more of the positions corresponding to V418, T420, V424, E505 and V547. 
     
     
         2 . The protein of  claim 1  wherein the VSA or the recombinant protein binds to FcRn at a pH in the range of 5.5 to 6.0, e.g., at a pH of 5.5 or 6.0, with higher affinity than a corresponding native serum albumin. 
     
     
         3 . The protein of  claim 1  or  2 , wherein the ratio of the binding affinity of a serum albumin comprising the VSA sequence at a pH of 5.5 to 6.0 to that at a pH of 7.0 to 7.4 is greater than or equal to the ratio for a corresponding native human albumin. 
     
     
         4 . The protein of any one of  claims 1 - 3 , wherein the ratio of the binding affinity of a serum albumin comprising the VSA sequence at a pH of 5.5 to 6.0 to that at a pH of 7.0 to 7.4 is 5; 10; 50; 100; 1000; 10,000; 100,000; or 1 million times that of a corresponding native human albumin. 
     
     
         5 . The protein of claim any one of  claims 1 - 4 , wherein a serum albumin comprising the VSA sequence binds to FcRn at a pH in the range of 7.0 to 7.4 with an affinity not greater than a corresponding native human albumin. 
     
     
         6 . The protein of claim any one of  claims 1 - 5  wherein the VSA sequence comprises a substitution of V418 with a methionine. 
     
     
         7 . The protein of any one of  claims 1 - 6 , wherein the VSA sequence comprises a substitution of T420 with an uncharged amino acid. 
     
     
         8 . The protein of  claim 7  wherein the VSA sequence comprises a substitution of T420 with alanine. 
     
     
         9 . The protein of any one of  claims 1 - 8 , wherein the VSA sequence comprises a substitution of V424 with an uncharged amino acid. 
     
     
         10 . The protein of  claim 9 , wherein the VSA sequence comprises a substitution of V424 with isoleucine. 
     
     
         11 . The protein of any one of  claims 1 - 10 , wherein the VSA sequence comprises a substitution of E505 with an uncharged amino acid or positively charged amino acid. 
     
     
         12 . The protein of  claim 11 , wherein the VSA sequence comprises a substitution of E505 with arginine, lysine or glycine. 
     
     
         13 . The protein of any one of  claims 1 - 12 , wherein the VSA sequence comprises a substitution of V547 with an uncharged amino acid. 
     
     
         14 . The protein of  claim 13 , wherein the VSA sequence comprises a substitution of V547 with alanine. 
     
     
         15 . The protein of any one of  claims 1 - 14 , wherein the VSA sequence comprises mutations at two or more of the positions selected from V418, T420, E505, and V547. 
     
     
         16 . The protein of  claim 15 , wherein the VSA sequence comprises two or more mutations selected from V418M, T420A, E505(R/K/G) and V547A. 
     
     
         17 . The protein of  claim 15 , wherein the VSA sequence comprises mutations at three or more positions selected from V418M, T420A, E505(R/K/G) and V547A. 
     
     
         18 . The protein of  claim 15 , wherein the VSA sequence comprises mutations V418M, T420A, E505(R/K/G) and V547A. 
     
     
         19 . The protein of any one of  claims 1 - 18 , wherein the VSA sequence comprises at least one mutation selected from V424I, N429D, M446V; A449V; T467M and A552T. 
     
     
         20 . The protein of any one of  claims 1 - 19 , wherein the VSA sequence is at least 80% identical but less than 100% identical to domain III of a naturally-occurring serum albumin. 
     
     
         21 . The protein of  claim 20  wherein the VSA sequence is at least 80% identical but less than 100% identical to the corresponding sequence of human serum albumin. 
     
     
         22 . The protein of any one of  claims 1 - 21 , wherein the protein comprises a heterologous sequence. 
     
     
         23 . The protein of  claim 22 , wherein the protein comprises a first and a second heterologous sequence. 
     
     
         24 . The protein of  claim 24  wherein the first and the second heterologous sequence are identical and are positioned in tandem. 
     
     
         25 . The protein of  claim 23  wherein the first heterologous sequence is located N-terminal to the variant sequence and the second heterologous sequence is located C-terminal to the variant sequence. 
     
     
         26 . The protein of  claim 22  wherein the heterologous sequence comprises a cytokine domain. 
     
     
         27 . The protein of  claim 26  wherein the cytokine is interleukin-2. 
     
     
         28 . The protein of  claim 22 , wherein the heterologous sequence comprises an immunoglobulin variable domain. 
     
     
         29 . The protein of  claim 22 , wherein the heterologous sequence comprises an Adnectin™, a DARPin, or an anti-calin, or a fragment of an Adnectin™, a DARPin, or an anti-calin, 
     
     
         30 . The protein of  claim 22 , wherein the heterologous sequence comprises a soluble fragment of a cell surface receptor. 
     
     
         31 . The protein of  claim 22 , wherein the heterologous sequence comprises an enzyme. 
     
     
         32 . The protein of  claim 22 , wherein the heterologous sequence comprises a functional fragment of a coagulation protein. 
     
     
         33 . The protein of  claim 32 , wherein the heterologous sequence comprises a functional fragment of FVII. 
     
     
         34 . The protein of  claim 32 , wherein the heterologous sequence comprises a functional fragment of FVIII. 
     
     
         35 . An isolated, recombinant protein comprising a VSA that has altered binding properties for human FcRn relative to a wild type human serum albumin and binds to FcRn with a K D  of less than 50 nM at pH 5.5 and optionally an affinity for FcRn at pH 7.4 that is less than or equal to the affinity for FcRn of a wild type albumin at pH 7.4. 
     
     
         36 . A method of treating a subject, the method comprising administering to the subject an effective amount of a therapeutic agent in association with the protein of any one of  claim 1 - 21  or  35 , such that the dosage and/or frequency of administration at which the agent produces a therapeutic effect is reduced relative to the dosage and/or frequency of administration at which the agent produces a therapeutic effect when it is not in association with the albumin protein. 
     
     
         37 . The method of  claim 36 , wherein the agent comprises a polypeptide component that is fused to the albumin protein. 
     
     
         38 . The method of  claim 37 , wherein the polypeptide component and the albumin protein are separated by a linker sequence. 
     
     
         39 . The method of  claim 37 , wherein the polypeptide component and the albumin protein are covalently linked by a non-peptide bond. 
     
     
         40 . The method of  claim 37 , wherein the polypeptide component and the albumin protein are non-covalently and stably associated. 
     
     
         41 . A method of engineering a VSA associated therapeutic agent, the method comprising:
 providing a biologically or pharmaceutically active agent; and   associating the agent with the protein of any one of  claim 1 - 21  or  35  to provide a VSA associated therapeutic agent.   
     
     
         42 . The method of  claim 41 , further comprising formulating the VSA associated therapeutic agent for administration to a subject. 
     
     
         43 . A method of engineering a VSA associated diagnostic agent, the method comprising:
 providing a diagnostic agent; and   associating the agent with the protein of any one of  claim 1 - 21  or  35  to provide a VSA associated diagnostic agent.   
     
     
         44 . The method of  claim 43  further comprising formulating the VSA associated diagnostic agent for administration to a subject. 
     
     
         45 . The method of  claim 44  further comprising administering the VSA associated diagnostic agent to a subject and detecting the VSA associated diagnostic agent. 
     
     
         46 . The method of  claim 45 , wherein the subject is imaged. 
     
     
         47 . A method of treating a subject with a VSA fusion protein, the fusion protein comprising a therapeutic agent linked to a VSA, wherein the VSA comprises a sequence that is a variant of domain III of a naturally-occurring serum albumin that comprises a mutation at one or more of the positions corresponding to V418, T420, V424, E505 and V547;
 the method comprising administering to the subject a therapeutically effective amount of the VSA fusion protein, such that the dosage and/or frequency of administration at which the agent produces a therapeutic effect is reduced relative to the dosage and/or frequency of administration at which the agent produces a therapeutic effect when it is not in association with the VSA.   
     
     
         48 . The method of  claim 47 , wherein the therapeutic agent comprises a sequence encoding human IL-2 or an active variant of IL-2. 
     
     
         49 . The method of  claim 48 , wherein the subject suffers from, or is at risk of suffering from, an immune disorder. 
     
     
         50 . The method of  claim 49 , wherein the subject has undergone, or plans to undergo, a procedure selected from the group consisting of an organ transplant, or blood transfusion, or bone marrow transplantation. 
     
     
         51 . The method of  claim 47 , wherein the therapeutic agent comprises a sequence encoding a urate oxidase, or an active variant of a urate oxidase. 
     
     
         52 . The method of  claim 51 , wherein the subject suffers from gout.

Join the waitlist — get patent alerts

Track US2014315817A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.