Anti-cd89 cytotoxic complex

Patent metadata

Field	Value
Publication number	US-2018050102-A1
Application number	US-201515560770-A
Country	US
Kind code	A1
Filing date	Mar 25, 2015
Priority date	Mar 25, 2015
Publication date	Feb 22, 2018
Grant date	—

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

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.

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 single domain, or a fragment thereof, 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, Ricin A, and a variant or functional fragment thereof.

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 . The complex according to claim 1 , wherein the binding structure comprises the amino acid sequence of SEQ ID NO: 1, or a recombinant or synthetic homologous peptide thereof, having an amino acid sequence which is at least 85%, optionally 90%, optionally 95% identical to the amino acid sequence of SEQ ID NO: 1 differing by substitution, insertion, addition or deletion of one or more amino acid residues, or fragments thereof.

6 . The complex according to claim 1 , 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, or recombinant or synthetic homologous polypeptides, variants or mutations thereof having an amino acid which is at least 85%, optionally 90%, optionally 95% identical to the amino acid sequence of SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4 or SEQ ID NO. 5, differing by substitution, insertion, addition or deletion of one or more amino acid residues, or fragments thereof.

7 . The complex according to claim 1 , wherein the human target cell is a cancer cell, in particular a malignant myeloid cell.

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

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

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

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

12 . A method for preparing a complex according claim 1 , the method comprising culturing the host cell according to claim 10 and isolating the complex from the cultured host cell.

13 . 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 11 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.

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

15 . 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.

16 . A polypeptide suitable for the detection of CD89, wherein said polypeptide comprises the amino acid sequence of SEQ ID NO: 1, or a recombination or synthetic homologous polypeptide, variant or mutation thereof having an amino acid which is at least 85%, optionally 90%, optionally 95% identical to the amino acid sequence of SEQ ID NO. 1, which differs by substitution, insertion, addition or deletion of one or more amino acid residues, or fragments thereof.

17 . The polypeptide according to claim 16 , wherein said polypeptide is coupled to a detectable label; optionally to a fusion tag protein, optionally SNAP-tag, CLIP-tag, Lumio tag or HaloTag.

18 . The polypeptide according to claim 16 , wherein the CD89 is presented on a human target cell or a fragment thereof.

19 . 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 16 under conditions that allow for formation of a complex between the polypeptide and CD89; and detecting the formation of the complex.

20 . A method for the detection of CD89 contained in a sample, the method comprising:
(a) contacting a sample with a polypeptide according to claim 16 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.

21 . A method for the diagnosis of CD89+ malignancies, comprising contacting a biological sample taken from a patient with a polypeptide according to claim 16 .

Assignees

• Fraunhofer Ges Forschung
• Rheinisch Westfaelische Technische Hochschule Aachen
• Univ Friedrich Alexander Er

Inventors

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

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P35/02

  specific for leukemia · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P29/00

  Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] · CPC title