Antibody fragments for detecting cancer and methods of use

• US 10,166,304 B2
• Filed: 07/10/2015
• Issued: 01/01/2019
• Est. Priority Date: 07/11/2014
• Status: Active Grant

• Current Assignee: Regents of The University of Minnesota (University of Minnesota)
• Original Assignee: Regents of The University of Minnesota (University of Minnesota)
• Inventors: Panyam, Jayanth, Kalscheuer, Stephen
• Primary Examiner(s): Aeder, Sean E
• Application Number:
  US15/325,696
• Publication Number:
  US 20170157275A1
• Time in Patent Office: 1,271 Days
• Field of Search: None
• CPC Class Codes: A61K 47/6855 the tumour determinant being from breast cancer cell
• CPC Class Codes: A61K 47/6865 the tumour determinant being from skin, nerves or brain cancer cell
• CPC Class Codes: A61K 47/6869 the tumour determinant being from a cell of the reproductive system: ovaria, uterus, testes, prostate
• CPC Class Codes: A61K 47/6929 the form being a nanoparticle, e.g. an immuno-nanoparticle
• CPC Class Codes: A61K 49/0039 Coumarin dyes
• CPC Class Codes: A61K 49/0093 Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
• CPC Class Codes: A61K 51/1027 against receptors, cell-surface antigens or cell-surface determinants
• CPC Class Codes: A61K 51/1051 the tumor cell being from breast, e.g. the antibody being herceptin
• CPC Class Codes: A61K 51/1066 the tumor cell being from skin
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/626 Diabody or triabody
• CPC Class Codes: C07K 2317/77 Internalization into the cell
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: G01N 2400/40 Glycosaminoglycans, i.e. GAG or mucopolysaccharides, e.g. chondroitin sulfate, dermatan sulfate, hyaluronic acid, heparin, heparan sulfate, and related sulfated polysaccharides
• CPC Class Codes: G01N 33/57415 of breast
• CPC Class Codes: G01N 33/5743 of skin, e.g. melanoma
• CPC Class Codes: G01N 33/57434 of prostate
• CPC Class Codes: G01N 33/6857 Antibody fragments

Abstract

The present invention relates to diagnostic and therapeutic agents comprising recombinant antibody fragments to bind a protein associated with cancer and methods of use of these diagnostic and therapeutic agents.

Claims

1. An immune reagent comprising a first scFv antibody fragment that specifically binds to membrane protein HSPG2 (Perlecan), wherein the first scFv antibody fragment is Clone-6 (SEQ ID NO:

2).

2. The immune reagent of claim 1, wherein the immune reagent is about 26-29 kDa.

3. The immune reagent of claim 1, further comprising a second scFv antibody fragment operably linked to the first scFv antibody fragment to form a diabody.

4. The immune reagent of claim 3, wherein both the first and second scFv antibody fragments are Clone-6 (SEQ ID NO:

2).

5. The immune reagent of claim 3, wherein the first and second antibody fragments are linked by means of a linker.

6. The immune reagent of claim 3, further comprising a poly-His tail operably linked to either the first or second antibody fragment.

9. A conjugate comprising the immune reagent of claim 1 conjugated to a detection agent and/or therapeutic agent.

10. The conjugate of claim 9, wherein the detection agent or therapeutic agent comprises a radionuclide.

11. The conjugate of claim 10, wherein the radionuclide is selected from Antimony-124, Antimony-125, Arsenic-74, Barium-103, Barium-140, Beryllium-7, Bismuth-206, Bismuth-207, Cadmium-109, Cadmium-115m, Calcium-45, Cerium-139, Cerium-141, Cerium-144, Cesium-137, Chromium-51, Cobalt-55, Cobalt-56, Cobalt-57, Cobalt-58, Cobalt-60, Cobalt-64, Copper-64, Copper-67, Erbium-169, Europium-152, Gallium-64, Gallium-68, Gadolinium-153, Gadolinium-157 Gold-195, Gold-199, Hafnium-175, Hafnium-175-181, Holmium-166, Indium-110, Indium-111, Iridium-192, Iron-55, Iron-59, Krypton-85, Lead-210, Manganese-54, Mercury-197, Mercury-203, Molybdenum-99, Neodymium-147, Neptunium-237, Nickel-63, Niobium-95, Osmium-185+191, Palladium-103, Platinum-195m, Praseodymium-143, Promethium-147, Protactinium-233, Radium-226, Rhenium-186, Rhenium-188, Rubidium-86, Ruthenium-103, Ruthenium-106, Scandium-44, Scandium-46, Selenium-75, Silver-110m, Silver-111, Sodium-22, Strontium-85, Strontium-89, Strontium-90, Sulfur-35, Tantalum-182, Technetium-99m, Tellurium-125, Tellurium-132, Thallium-204, Thorium-228, Thorium-232, Thallium-170, Tin-113, Tin-114, Tin-117m, Titanium-44, Tungsten-185, Vanadium-48, Vanadium-49, Ytterbium-169, Yttrium-86, Yttrium-88, Yttrium-90, Yttrium-91, Zinc-65, and Zirconium-95.

12. The conjugate of claim 9, wherein the immune reagent is conjugated to a detection agent.

13. The conjugate of claim 12, wherein the detection agent comprises a fluorescent group.

14. The conjugate of claim 13, wherein the fluorescent group is fluorescein, tetrachlorofluorescein, hexachlorofluorescein, tetramethylrhodamine, rhodamine, cyanine-derivative dyes, Texas Red, Bodipy, and/or Alexa dye.

15. The conjugate of claim 9, wherein the immune reagent is conjugated to a therapeutic agent.

16. The conjugate of claim 15, wherein the therapeutic agent is a cytotoxic compound.

17. The conjugate of claim 16, wherein the cytotoxic compound is a chemotherapeutic agent.

18. The conjugate of claim 17, wherein the chemotherapeutic agent is selected from all-trans retinoic acid, Azacitidine, Azathioprine, Bleomycin, Bortezomib, Carboplatin, Capecitabine, Cisplatin, Chlorambucil, Cyclophosphamide, Cytarabine, Daunorubicin, Docetaxel, Doxifluridine, Doxorubicin, Epirubicin, Epothilone, Etoposide, Fluorouracil, Gemcitabine, Hydroxyurea, Idarubicin, Imatinib, Mechlorethamine, Mercaptopurine, Methotrexate, Mitoxantrone, Oxaliplatin, Paclitaxel, silicate prodrug of Paclitaxel, Pemetrexed, Teniposide, Tioguanine, Valrubicin, Vinblastine, Vincristine, Vindesine, Vinorelbine, Axitinib, Bosutinib, Cediranib, Dasatinib, Erlotinib, Gefitinib, Imatinib, Lapatinib, Lestaurtinib, Nilotinib, Semaxanib, Sunitinib, Vemurafinib and/or Vandetanib.

19. A pharmaceutical composition comprising the immune reagent of claim 1 and a pharmaceutically acceptable excipient.

20. A composition comprising an immune agent of claim 1 operably linked to a carrier.

21. The composition of claim 20 wherein the carrier is a nanoparticle or liposome.

22. A method of inhibiting proliferation and/or growth of a tumor by administering the immune reagent of claim 1 to a patient in need thereof.

24. A method of inhibiting proliferation and/or growth of a tumor by administering the conjugate of claim 9 to a patient in need thereof.

7. An immune reagent comprising

(a) a heavy chain encoded by a nucleic acid having 100% identity to SEQ ID NO:

3 and a light chain encoded by a nucleic acid having 100% identity to SEQ ID NO;

5;

or

(b) a heavy chain variable region having 100% identity to SEQ ID NO;

4 and a light chain variable region having 100% identity to SEQ ID NO;

6.

8. An immunoglobulin comprising a first immune reagent of claim 7 operably linked to a second immune reagent of claim 7.

23. A method of inhibiting proliferation and/or growth of a tumor by administering the immune reagent of claim 7 to a patient in need thereof.