GLUT-1 as a receptor for HTLV envelopes and its uses

Patent metadata

Field	Value
Publication number	US-9777044-B2
Application number	US-201414575132-A
Country	US
Kind code	B2
Filing date	Dec 18, 2014
Priority date	May 2, 2003
Publication date	Oct 3, 2017
Grant date	Oct 3, 2017

Abstract

The present application relates to polypeptides derived from the soluble part of the glycoprotein of the enveloped virus of Primate T-cell leukemia virus (PTLV), or fragments or variants thereof named receptor binding domain ligands (RBD) selected for their ability to bind specifically to the nutrient transporter GLUT1.

First claim

The invention claimed is:

1. An isolated fusion polypeptide, wherein said fusion polypeptide consist of a soluble receptor binding domain (RBD) ligand derived from the soluble part of the glycoprotein of the human T-cell leukemia virus (HTLV-2) binding to the Glucose Transporter 1 (GLUT1), consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 43, and fragments of SEQ ID NO: 4 or SEQ ID NO: 43,
wherein said fragments of SEQ ID NO: 43 are polypeptides delimited in their N-terminal extremity by the amino acid located in position 1 to 90 and in their C-terminal extremity by the amino acid located in position 135 to 245 of SEQ ID NO: 43; and
wherein said fragments of SEQ ID NO: 4 consist of amino acids 19 to 224 of SEQ ID NO: 4, or 20 to 224 of SEQ ID NO: 4, or 21 to 224 of SEQ ID NO: 4, or 19 to 178 of SEQ ID NO: 4, or 20to 178 of SEQ ID NO: 4, or 21 to 178 of SEQ ID NO: 4, and
wherein said isolated polypeptide is chemically modified to add a fluorochrome or a fluorescent compound.

2. The isolated polypeptide according to claim 1 , wherein said polypeptide binds to the Glucose Transporter 1 (GLUT1) comprising the amino acid sequence SEQ ID NO: 2 or to a fragment of GLUT1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 35 to SEQ ID NO: 42.

3. The isolated polypeptide according to claim 1 , wherein the fluorochrome or fluorescent compound is selected from the group consisting of Cyanine dye, Alexa dye and Quantum dye.

4. A method for diagnosing a GLUT1 related disease comprising:
collecting a biological sample from a subject,
determining the level of GLUT1 expression at a cell surface using the isolated polypeptide of claim 1 , and
comparing said level to a reference value for GLUT1 expression at the cell surface in a biological sample from a healthy individual, wherein an elevated level or decreased level of GLUT1 expression in the sample from the subject indicates a GLUT1 related disease.

5. The method according to claim 4 , wherein said GLUT1 related disease is GLUT1 deficiency syndrome.

6. The method according to claim 4 , wherein said GLUT1 related disease is a cancer disease.

7. An isolated fusion polypeptide, wherein said fusion polypeptide consists of:
a soluble receptor binding domain (RBD) ligand derived from the soluble part of the glycoprotein of the human T-cell leukemia virus (HTLV-2) binding to the Glucose Transporter 1 (GLUT1), consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 43, and fragments of SEQ ID NO: 4 or SEQ ID NO: 43,
wherein said fragments of SEQ ID NO: 43 are polypeptides delimited in their N-terminal extremity by the amino acid located in position 1 to 90 and in their C-terminal extremity by the amino acid located in position 135 to 245 of SEQ ID NO: 43, and
wherein said fragments of SEQ ID NO: 4 consist of amino acids 19 to 224 of SEQ ID NO: 4, or 20 to 224 of SEQ ID NO: 4, or 21 to 224 of SEQ ID NO: 4, or 19 to 178 of SEQ ID NO: 4, or 20 to 178 of SEQ ID NO: 4, or 21 to 178 of SEQ ID NO: 4; and

a Tag, an antibody constant fragment, or a fluorescent protein.

8. The isolated fusion polypeptide according to claim 7 , wherein the tag is selected from the group consisting of hemaglutinin tag, poly arginine tag, poly histidine tag, myc tag, strep tag, S-tag, HAT tag, 3 times flag tag, calmodulin-binding peptide tag, SBP tag, chitin binding domain tag, GST tag, maltose-binding protein tag, fluorescent protein tag, T7 tag, V5tag and Xpress tag.

9. The isolated fusion polypeptide according to claim 7 , wherein the antibody constant fragment is selected from the group consisting of Fc fragment from rabbit and Fc fragment from mouse.

10. The isolated fusion polypeptide according to claim 7 , wherein the fluorescent protein is GFP.

11. The isolated fusion polypeptide according to claim 7 , wherein said polypeptide binds to the Glucose Transporter 1 (GLUT1) comprising the amino acid sequence SEQ ID NO: 2, or to a fragment of GLUT1 having an amino acid sequence selected from the group consisting of SEQ ID NO: 35 to SEQ ID NO: 42.

12. A kit of parts comprising at least one isolated fusion polypeptide, wherein said fusion polypeptide is the fusion polypeptide of claim 7 .

13. A method for diagnosing a GLUT1 related disease comprising:
collecting a biological sample from a subject,
determining the level of GLUT1 expression at a cell surface using the isolated polypeptide of claim 7 , and
comparing said level to a reference value for GLUT1 expression at the cell surface in a biological sample from a healthy individual, wherein an elevated level or decreased level of GLUT1 expression in the sample from the subject indicates a GLUT1related disease.

14. The method according to claim 13 , wherein said GLUT1 related disease is GLUT1 deficiency syndrome.

15. The method according to claim 13 , wherein said GLUT1 related disease is a cancer disease.

16. A composition comprising at least one isolated fusion polypeptide, wherein said fusion polypeptide consists of:
a soluble receptor binding domain (RBD) ligand derived from the soluble part of the glycoprotein of the human T-cell leukemia virus (HTLV-2) binding to the Glucose Transporter 1 (GLUT1), consisting of the amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 43, and fragments of SEQ ID NO: 4 or SEQ ID NO: 43,
wherein said fragments of SEQ ID NO: 43 are polypeptides delimited in their N-terminal extremity by the amino acid located in position 1 to 90 and in their C-terminal extremity by the amino acid located in position 135 to 245 of SEQ ID NO: 43; and
wherein said fragments of SEQ ID NO: 4 consist of amino acids 19 to 224 of SEQ ID NO: 4, or 20 to 224 of SEQ ID NO: 4, or 21 to 224 of SEQ ID NO: 4, or 19 to 178 of SEQ ID NO: 4, or 20 to 178 of SEQ ID NO: 4, or 21 to 178 of SEQ ID NO: 4; and

a Tag, an antibody constant fragment, or a fluorescent protein.

17. The composition according to claim 16 , being a pharmaceutical composition and further comprising a pharmaceutically acceptable excipient.

Assignees

• Centre Nat De La Rech Scient (Cnrs)
• Univ De Montpellier 2

Inventors

• Battini Jean-Luc
• Manel Nicolas
• Kim Felix
• Kinet Sandrina
• Taylor Naomi
• Sitbon Marc

Classifications

• C07K14/62

  Insulins · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• G01N2333/62

  Insulins · CPC title

• C12N2740/14022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C12N2740/15022

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title