CTLA-4 Variants
AssigneeMedImmune, LLC
InventorsRalph Minter, Julie Douthwaite, Jacques Moisan, Michael Bowen, Steven Rust, Cyril Privezentzev
Life of this patent
Estimating the term…
Abstract
Variants of cytotoxic T-lymphocyte antigen 4 (CTLA-4) with high affinity, potency and stability. Formulations of CTLA-4 variants at high concentration for subcutaneous or intravenous administration, e.g. at monthly or less frequent dosage intervals. Use of CTLA-4 variants for treating rheumatoid arthritis and other inflammatory disorders. Fusion of CTLA-4 with IgG Fc having improved stability and longer in vivo half-life.
Claims51
An isolated CTLA-4 polypeptide having greater affinity for binding human CD80, greater potency and/or greater stability compared with wild type CTLA-4 SEQ ID NO:
- 35, the polypeptide comprising an amino acid sequence that is a variant of SEQ ID NO:
- 35, wherein the variant comprises five or more of the following amino acid mutations in SEQ ID NO:
- 35:
- R, S, V or T at I16;
- T at A24;
- N or P at S25;
- S at G27;
- I at V 32;
- G at D41;
- G at S42;
- E at V44;
- K at M54;
- S or G at N56;
- A, G, S or P at L58;
- S or A at T59;
- T at F 60;
- Q or P at L61;
- G at D 62;
- Y at D63;
- P at S 64;
- N, D, V or T at I65;
- A, T, M or H at S70;
- R at Q80;
- Q, S, V, R, K or L at M85;
- S at T87;
- Q, H, T, E or M at K93;
- R, Q or E at L104;
- V at I106;
- D or S at N108;
- V or F at I115;
- S at C120;
- deletion at T51.
A CTLA-4 polypeptide according to any of the preceding claims, comprising an amino acid sequence selected from SEQ ID NOS:
- 36-55 or comprising the CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948, or comprising a variant of any of those sequences with up to ten amino acid mutations.
A CTLA-4 polypeptide according to any of the preceding claims, comprising an amino acid sequence selected from SEQ ID NOS:
- 36-55 or comprising the CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948, or comprising a variant of any of those sequences with up to five amino acid mutations.
An isolated CTLA-4 polypeptide having greater affinity for binding human CD80, greater potency and/or greater stability compared with wild type CTLA-4 SEQ ID NO:
- 35, wherein the polypeptide comprises:
- (i) amino acid sequence SEQ ID NO:
- 68, SEQ ID NO:
- 43, SEQ ID NO:
- 37, SEQ ID NO:
- 38, SEQ ID NO:
- 36, SEQ ID NO:
- 42 or SEQ ID NO:
- 47;
- (ii) an amino acid sequence that is a variant of (i) containing up to ten amino acid mutations, wherein residue 25 is not mutated and is N;
- (iii) an amino acid sequence that is a variant of (i) comprising one or more amino acid mutations, wherein residue 25 is not mutated and is N, the variant having at least 70% sequence identity to (i);
- or
- (iv) a CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948.
A CTLA-4 polypeptide according to any of the preceding claims, having an affinity of 50 nM or less for binding human CD80, wherein affinity is KD as determined by surface plasmon resonance.
A CTLA-4 polypeptide according to any of the preceding claims, wherein the polypeptide has greater affinity than wild type CTLA-4 (SEQ ID NO:
- 35) for binding human CD86.
An isolated CTLA-4 polypeptide which has at least 10-fold greater affinity for binding CD80 than for binding CD86.
A CTLA-4 polypeptide according to any of the preceding claims, conjugated to an IgG Fc amino acid sequence.
An isolated CTLA-4 polypeptide comprising the 1299 CTLA-4-Ig amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948.
A CTLA-4 polypeptide according to any of the preceding claims, wherein the polypeptide is in a multimer.
A host cell containing nucleic acid, wherein the nucleic acid comprises the 1299 CTLA-4-Ig nucleic acid sequence deposited under NCIMB accession no. 41948.
A composition comprising:
- a CTLA-4 polypeptide according to any of the preceding claims;
- and
- one or more pharmaceutical excipients.
A composition comprising:
- a CTLA-4 polypeptide comprising amino acid sequence SEQ ID NO:
- 68, SEQ ID NO:
- 43, SEQ ID NO:
- 47, SEQ ID NO:
- 38, SEQ ID NO:
- 36, SEQ ID NO:
- 42 SEQ ID NO:
- 47 or the CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948, conjugated to an IgG Fc amino acid sequence;
- and
- one or more pharmaceutical excipients.
A composition comprising the 1299 CTLA-4-Ig polypeptide encoded by nucleic acid deposited under NCIMB accession no. 41948 and one or more pharmaceutical excipients.
A method of producing a further CTLA-4 polypeptide by mutation of a CTLA-4 polypeptide amino acid sequence selected from SEQ ID NOS 36-55 or the CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948, the method comprising:
- providing a CTLA-4 polypeptide comprising or consisting of amino acid sequence SEQ ID NOS 36-55 or the CTLA-4 amino acid sequence encoded by nucleic acid deposited under NCIMB accession no. 41948;
- introducing one or more mutations in the amino acid sequence to provide a further CTLA-4 polypeptide;
- testing stability, affinity and/or potency of the further CTLA-4 polypeptide;
- and
- formulating the further CTLA-4 polypeptide into a composition comprising one or more pharmaceutical excipients.
Dependencies are inferred from the claim text.