Anti-CD89 cytotoxic complex

Patent metadata

Field	Value
Publication number	US-10668147-B2
Application number	US-201515560770-A
Country	US
Kind code	B2
Filing date	Mar 25, 2015
Priority date	Mar 25, 2015
Publication date	Jun 2, 2020
Grant date	Jun 2, 2020

Abstract

Complexes suitable for targeting and killing a human target cell comprising a first polypeptide with a binding structure for binding the complex to the cellular surface receptor CD89 (e.g. with an anti CD89 antibody) presented on the cell surface of the human target cell and a second polypeptide comprising a toxic effector domain, preferably Pseudomonas exotoxin A); to nucleic acid molecules encoding said complexes, vectors, host cells containing the nucleic acids and methods for preparation and producing such complexes; compositions and methods for using said complexes for the treatment of diseases, in particular of cancer diseases like leukemia.

First claim

What is claimed is:

1. A complex suitable for targeting and killing a human target cell, the complex comprising a first polypeptide comprising a binding structure for binding the complex to a cellular surface receptor CD89 presented on the cell surface of said human target cell and a second polypeptide comprising a toxic effector domain, wherein the first polypeptide comprises the amino acid sequence of SEQ ID NO: 1.

2. The complex according to claim 1 , wherein the binding structure is an antibody or an antibody fragment selected from the group consisting of an Fab, a scFv, a bis scFv, an Fab 2 , an Fab 3 , a minibody, a diabody, a triabody, a tetrabody and a tandab.

3. The complex according to claim 1 , wherein the toxic effector domain is selected from the group consisting of a protease, a serine protease, granzyme B, granzyme A, granzyme H, granzyme K, granzyme M, a trypsin, a chymotrypsin, a bacteria-originated toxic compound, Pseudomonas aeruginosa exotoxin A (ETA), a human hydrolase, angiogenin, a cytoskeleton-associated protein, microtubule-associated protein tau, a photosensitizer, a plant-originated toxin, and Ricin A.

4. The complex according to claim 1 , wherein the binding structure is a CD89-specific single-chain variable fragment (scFv) and the toxic effector domain is selected from the group consisting of Pseudomonas aeruginosa exotoxin A (ETA), granzyme B, angiogenin, and microtubule-associated protein tau.

5. A complex suitable for targeting and killing a human target cell, the complex comprising a first polypeptide comprising a binding structure for binding the complex to a cellular surface receptor CD89 presented on the cell surface of said human target cell and a second polypeptide comprising a toxic effector domain, wherein the complex comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5.

6. The complex according to claim 1 , wherein the human target cell is a cancer cell.

7. An isolated nucleic acid molecule encoding the complex according to claim 1 .

8. A vector comprising the nucleic acid molecule of claim 7 .

9. A host cell transformed with the vector of claim 8 .

10. A pharmaceutical composition comprising a complex according to claim 1 in combination with a pharmacologically acceptable carrier, diluent, stabilizer or formulation.

11. A method for preparing a complex suitable for targeting and killing a human target cell, the complex comprising a first polypeptide comprising a binding structure for binding the complex to a cellular surface receptor CD89 presented on the cell surface of said human target cell and a second polypeptide comprising a toxic effector domain, the method comprising culturing the host cell according to claim 10 and isolating the complex from the cultured host cell.

12. A method for the treatment of a disease selected from the group consisting of a malignant disease, a chronic inflammatory disease, a cutaneous disease, an autoimmune disease, and an intestinal disease, the method comprising administering an effective amount of the pharmaceutical of claim 10 to a patient in need thereof, wherein:
the malignant disease or chronic inflammatory disease is selected from the group consisting of acute myeloid leukaemia, arthritis, chronic obstructive pulmonary disease (COPD), emphysema, intrinsic asthma, and extrinsic asthma;
the cutaneous disease is selected from the group consisting of atopic dermatitis, psoriasis, polymorphic light eruption, and systemic lupus erythematosus (SLE);
the autoimmune disease is selected from the group consisting of graft versus host, multiple sclerosis, macrophage activation syndrome, rheumatoid arthritis, juvenile arthritis;
the intestinal disease is Crohn's disease or chronic bowel disease.

13. The method according to claim 12 , wherein the disease is myeloid leukaemia.

14. A method of treating a malignant disease, autoimmune disease, tissue rejection reaction, or chronic inflammatory disease comprising administering an effective amount of the complex according to claim 1 to a patient in need thereof.

15. A polypeptide suitable for the detection of CD89, wherein said polypeptide comprises the amino acid sequence of SEQ ID NO: 1.

16. The polypeptide according to claim 15 , wherein said polypeptide is coupled to a detectable label.

17. The polypeptide according to claim 15 , wherein the CD89 is presented on a human target cell or a fragment thereof.

18. A method of detecting the presence of CD89 or a cell expressing CD89 in a sample, comprising: contacting the sample with a polypeptide according to claim 15 under conditions that allow for formation of a complex between the polypeptide and CD89; and detecting the formation of the complex.

19. A method for the detection of CD89 contained in a sample, the method comprising:
(a) contacting a sample with a polypeptide according to claim 15 and with a fusion tag specific fluorophore that specifically binds the fusion tag coupled to said polypeptide; and
(b) detecting the presence of CD89 in the sample by fluorescence signals associated with the fusion tag specific fluorophore.

20. A method for the diagnosis of CD89+ malignancies, comprising contacting a biological sample taken from a patient with a polypeptide according to claim 15 .

Assignees

• Fraunhofer Ges Forschung
• Rheinisch Westfaelische Technische Hochschule Aachen

Inventors

• Stein Christoph
• Mladenov Radoslav
• Stockmeyer Bernhard
• Barth Stefan
• Schenke Lea Christin
• Fischer Rainer

Classifications

• G01N33/574

  Physics · mapped topic

• G01N33/566 Primary

  using specific carrier or receptor proteins as ligand binding reagents {where possible specific carrier or receptor proteins are classified with their target compounds} · CPC title

• A61K39/395 Primary

  Antibodies (agglutinins A61K38/36 {; as drug carriers A61K47/50}); Immunoglobulins; Immune serum, e.g. antilymphocytic serum · CPC title

• C07K16/28

  against receptors, cell surface antigens or cell surface determinants · CPC title

• A61K47/6809

  Antibiotics, e.g. antitumor antibiotics anthracyclins, adriamycin, doxorubicin or daunomycin · CPC title