Insulin-like growth factor 1 receptor-specific antibodies and uses thereof

Patent metadata

Field	Value
Publication number	US-10100117-B2
Application number	US-201415123735-A
Country	US
Kind code	B2
Filing date	Dec 4, 2014
Priority date	Mar 6, 2014
Publication date	Oct 16, 2018
Grant date	Oct 16, 2018

Abstract

The blood-brain barrier (BBB) prevents transport of molecules larger than 500 Dal tons from blood to brain. Receptor-mediated transcytosis (RMT) facilitates transport across the BBB of specific molecules that bind receptors on brain endothelial cells that form the BBB. An insulin-like growth factor 1 receptor (IGF 1R)-binding antibody or fragment thereof is identified that transmigrates the BBB by RMT. The antibody or fragment is used to deliver a cargo molecule across the BBB, wherein the cargo molecule may be a therapeutic or detectable agent. The antibody is a camelid VHH, prepared by immunizing a llama with a 933-amino acid IGF 1R polypeptide. Humanized forms of the camelid VHH are also generated.

First claim

The invention claimed is:

1. An isolated or purified single domain antibody or antigen-binding fragment thereof, comprising
a complementarity determining region (CDR) 1 sequence of GRTIDNYA (SEQ ID NO:1);
a CDR2 sequence of IDWGDGGX (SEQ ID NO:2), where X is A or T; and
a CDR3 sequence of AMARQSRVNLDVARYDY (SEQ ID NO:3),
wherein the single domain antibody or antigen-binding fragment thereof specifically binds the Insulin-Like Growth Factor 1 Receptor (IGF1R).

2. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , comprising the sequence X 1 VX 2 LX 3 ESGGGLVQX 4 GGSLRLSCAASGRTIDNYAMAWX 5 RQAPGKX 6 X 7 EX 8 V X 9 TIDWGDGGX 10 RYANSVKGRFTISRDNX 11 KX 12 TX 13 YLQMNX 14 LX 15 X 16 EDTAV YX 17 CAMARQSRVNLDVARYDYWGQGTX 18 VTVSS (SEQ ID NO:4), where X 1 is E or Q; X 2 is K or Q; X 3 is V or E; X 4 is A or P; X 5 is V or S; X 6 is D or G; X 7 is L or R; X 8 is F or W; X 9 is A or S; X 10 is A or T; X 11 is A or S; X 12 is G or N; X 13 is M or L; X 14 is N or R; X 15 is E or R; X 16 is P or A; X 17 is S or Y; and X 18 is Q or L.

3. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , comprising a sequence selected from the group consisting of:

(SEQ ID NO: 5)

QVKLEESGGGLVQAGGSLRLSCAASGRTIDNYAMAWSRQAPGKDREFVA

TIDWGDGGARYANSVKGRFTISRDNAKGTMYLQMNNLEPEDTAVYSCAM

ARQSRVNLDVARYDYWGQGTQVTVSS;

(SEQ ID NO: 6)

EVQLVESGGGLVQPGGSLRLSCAASGRTIDNYAMAWVRQAPGKGLEWVS

TIDWGDGGTRYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAM

ARQSRVNLDVARYDYWGQGTLVTVSS;

(SEQ ID NO: 7)

QVQLVESGGGLVQPGGSLRLSCAASGRTIDNYAMAWVRQAPGKGLEWVA

TIDWGDGGTRYANSVKGRFTISRDNSKNTMYLQMNSLRAEDTAVYYCAM

ARQSRVNLDVARYDYWGQGTLVTVSS;

(SEQ ID NO: 8)

QVQLVESGGGLVQPGGSLRLSCAASGRTIDNYAMAWSRQAPGKGLEFVA

TIDWGDGGTRYANSVKGRFTISRDNSKNTMYLQMNSLRAEDTAVYYCAM

ARQSRVNLDVARYDYWGQGTLVTVSS;

(SEQ ID NO: 9)

QVQLVESGGGLVQPGGSLRLSCAASGRTIDNYAMAWSRQAPGKDREFVA

TIDWGDGGTRYANSVKGRFTISRDNSKGTMYLQMNSLRAEDTAVYSCAM

ARQSRVNLDVARYDYWGQGTLVTVSS; 

and

(SEQ ID NO: 10)

EVQLVESGGGLVQPGGSLRLSCAASGRTIDNYAMAWSRQAPGKDREFVS

TIDWGDGGTRYANSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAM

ARQSRVNLDVARYDYWGQGTLVTVSS.

4. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody is of camelid origin.

5. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding antibody fragment thereof is in a multivalent display format.

6. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 5 , wherein the single domain antibody or antigen-binding fragment thereof is linked to a Fc fragment.

7. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding fragment thereof is immobilized onto a surface.

8. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 , wherein the single domain antibody or antigen-binding fragment thereof is linked to a cargo molecule.

9. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 8 , wherein the cargo molecule has a molecular weight in the range of about 1 kD to about 200 kDa.

10. The isolated or purified single domain antibody or antigen-binding fragment thereof of claim 8 , wherein the cargo molecule is a detectable agent, a therapeutic, a peptide, a growth factor, a cytokine, a receptor trap, a chemical compound, a carbohydrate moiety, an enzyme, an antibody or fragment thereof, a DNA-based molecule, a viral vector, or a cytotoxic agent; one or more liposomes or nanocarriers loaded with a detectable agent, a therapeutic, a peptide, an enzyme, an antibody or fragment thereof, a DNA-based molecule, a viral vector, or a cytotoxic agent; or one or more nanoparticle, nanowire, nanotube, or quantum dots.

11. A composition comprising one or more than one isolated or purified single domain antibody or antigen-binding fragment thereof of claim 1 and a pharmaceutically-acceptable carrier, diluent, or excipient.

Assignees

• Nat Res Council Canada

Inventors

• Stanimirovic Danica
• KEMMERICH Kristin
• Haqqani Arsalan S
• Sulea Traian
• Arbabi-Ghahroudi Mehdi
• Massie Bernard
• GILBERT Rénald

Classifications

• G01N33/5759

  involving compounds localised on the membrane of tumour or cancer cells · CPC title

• A61K47/6811 Primary

  the drug being a protein or peptide, e.g. transferrin or bleomycin · CPC title

• C07K2317/77

  Internalization into the cell · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title