CXCR4 binding molecules and methods of use thereof
The present disclosure relates to polypeptides (also referred to herein as CXCR4 binding molecules or polypeptides) that are directed against the G-coupled protein receptor CXCR4, also known as Fusin or CD184. The invention also relates to nucleic acids encoding such polypeptides; to methods for preparing such polypeptide; to compositions, and in particular to pharmaceutical compositions that comprise such polypeptides and to uses of such polypeptides for therapeutic or diagnostic purposes.
1. A polypeptide which binds to human C-X-C chemokine receptor type 4 (CXCR4), comprising a scaffold region and binding loop regions contained therein, the scaffold region comprising a sequence which has at least 90% identity to the scaffold region defined by amino acids 1 to 26, 33 to 79 and 88 to 97 of SEQ ID NO:1; and
wherein the first binding loop region comprises a sequence represented by the formula SX 1 SX 2 X 3 R wherein
X 1 is G, K, L or Y;
X 2 is D, H, G or N; and
X 3 is I, V, M, F, Q or Y; or
a conservative amino acid substitution thereof
and/or
wherein the second binding loop region comprises a sequence represented by the formula Y′ 1 RY′ 2 GY′ 3 YRHRY′ 4 LY′ 5 LG wherein
Y′ 1 is Y or W;
Y′ 2 is T, V or I;
Y′ 3 is G or A;
Y′ 4 is A or Y; and
Y′ 5 is V, R or K; or
a conservative amino acid substitution thereof.
2. The polypeptide according to claim 1 , wherein the first binding loop region comprises or consists of the sequence set forth in any one of SEQ ID NO: 41, SEQ ID NO: 45, SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO:57, SEQ ID NO:61, SEQ ID NO:65, SEQ ID NO:69, SEQ ID NO: 73, or SEQ ID NO:77; and/or
wherein the second binding loop region comprises or consists of the sequence set forth in any one of SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, SEQ ID NO:62, SEQ ID NO:66, SEQ ID NO:70, SEQ ID NO: 74, or SEQ ID NO:78.
3. The polypeptide according to claim 1 or 2 :
(i) comprising a sequence at least 90% identical to the sequence selected from the group consisting of SEQ ID NO: 40, SEQ ID NO: 44, SEQ ID NO: 48, SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 72 or SEQ ID NO: 76; or
(ii) comprising or consisting of the sequence set forth in any one of SEQ ID NO: 40, SEQ ID NO: 44, SEQ ID NO: 48, SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, SEQ ID NO: 64, SEQ ID NO: 68, SEQ ID NO: 72 or SEQ ID NO: 76.
4. The polypeptide according to claim 1 which binds to one or more residues of CXCR4 selected from the group consisting of C28, E32, V112, Y184, F189, P191, D193, W195, D262, L266, E268 and E288 of human CXCR4 (SEQ ID NO:105).
5. A conjugate comprising a polypeptide according to claim 1 and an agent selected from a therapeutic agent, a toxin, a detectable label or an agent which extends the half-life of the polypeptide.
6. A polypeptide according to claim 1 , or the conjugate of claim 5 , for use in the treatment or to reduce the likelihood of occurrence of a CXCR4-related disease or disorder, wherein the disease or disorder is selected from the group consisting of cancer, inflammatory disease and fibrosis.
7. A pharmaceutical composition comprising the polypeptide according to claim 1 and an acceptable carrier.
8. An expression construct comprising the nucleic acid molecule encoding the polypeptide according to claim 1 .
9. A method of reducing the likelihood of occurrence of or treating a CXCR4-related disease or disorder wherein the disease or disorder is selected from the group consisting of cancer, inflammatory diseases and fibrosis comprising administering to a subject in need thereof a polypeptide according to claim 1 .
10. A method for localizing and/or detecting and/or diagnosing a CXCR4-related disease or disorder, the method comprising detecting in vivo the polypeptide according to claim 1 bound to CXCR4, if present, wherein the polypeptide is conjugated to a detectable tag.