Claims (22)
1. Dijatijelo sposobno za specifično vezanje za epitop CD123 i za epitop CD3, naznačeno time da dijatijelo sadrži prvi polipeptidni lanac i drugi polipeptidni lanac, kovalentno povezani jedan s drugim, pri čemu:
A. prvi polipeptidni lanac sadrži, u smjeru od N-završetka do C-završetka:
i. domena 1, koja sadrži
(1) poddomenu (1A), koja sadrži VL domenu monoklonskog antitijela sposobnog vezati za CD3 (VLCD3) i koja ima aminokiselinsku sekvencu SEK ID BR: 21; i
(2) poddomenu (1B), koja sadrži VH domenu monoklonskog antitijela sposobnu vezati za CD123 (VHCD123) i koja ima aminokiselinsku sekvencu SEK ID BR: 26, pri čemu su navedene poddomene 1A i 1B međusobno odvojene peptidnim poveznikom koji ima aminokiselinsku sekvencu SEK ID BR: 29;
ii. domena 2, pri čemu je navedena domena 2 domena E-spirale koja ima aminokiselinsku sekvencu SEK ID BR: 34; ili domenu K-spirale koja ima aminokiselinsku sekvencu SEK ID BR: 35, pri čemu je navedena domena 2 odvojena od navedene domene 1 pomoću peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 30; i
B. drugi polipeptidni lanac sadrži, u smjeru od N-završetka do C-završetka:
i. domenu 1, koja sadrži
(1) poddomenu (1A), koja sadrži VL domenu monoklonskog antitijela sposobnu povezati za CD123 (VLCD123) i koja ima aminokiselinsku sekvencu SEK ID BR: 25; i
(2) poddomena (1B), koja sadrži VH domenu monoklonskog antitijela sposobnu povezati na CD3 (VHCD3) i koja ima aminokiselinsku sekvencu SEK ID BR: 22, pri čemu su spomenute poddomene 1A i 1B međusobno odvojene peptidnim poveznikom koji ima aminokiselinsku sekvencu SEK ID BR: 29;
ii. domena 2, pri čemu je navedena domena 2 domena K-spirale koja ima aminokiselinsku sekvencu SEK ID BR: 35; i ili domena E-zavojnice koja ima aminokiselinsku sekvencu SEK ID BR: 34, pri čemu je navedena domena 2 odvojena od navedene domene 1 peptidnim poveznikom koja ima aminokiselinsku sekvencu SEK ID BR: 30; i pri čemu navedena domena 2 spomenutog prvog i spomenutog drugog polipeptidnog lanca nisu obje domene E-zavojnice ili obje domene K-zavojnice,
i pri čemu:
(a) spomenuta VL domena spomenutog prvog polipeptidnog lanca i spomenuta VH domena spomenutog drugog polipeptidnog lanca stvaraju antigen vežući domenu sposobnu za specifično vezanje za epitop CD3; i
(b) spomenuta VL domena spomenutog drugog polipeptidnog lanca i navedena VH domena spomenutog prvog polipeptidnog lanca stvaraju antigen vežući domenu sposobnu za specifično vezanje za epitop CD123.1. A dibody capable of specific binding to the CD123 epitope and to the CD3 epitope, characterized in that the dibody contains a first polypeptide chain and a second polypeptide chain, covalently linked to each other, wherein:
A. the first polypeptide chain contains, in the direction from the N-terminus to the C-terminus:
i. domain 1, which contains
(1) subdomain (1A), which contains the VL domain of a monoclonal antibody capable of binding to CD3 (VLCD3) and which has the amino acid sequence of SEQ ID NO: 21; and
(2) subdomain (1B), which contains the VH domain of a monoclonal antibody capable of binding to CD123 (VHCD123) and which has the amino acid sequence of SEQ ID NO: 26, wherein said subdomains 1A and 1B are separated from each other by a peptide linker having the amino acid sequence of SEK ID NO: 29;
ii. domain 2, wherein said domain 2 is an E-helix domain having the amino acid sequence SEQ ID NO: 34; or a K-helix domain having the amino acid sequence of SEQ ID NO: 35, wherein said domain 2 is separated from said domain 1 by a peptide linker having the amino acid sequence of SEQ ID NO: 30; and
B. the second polypeptide chain contains, in the direction from the N-terminus to the C-terminus:
i. domain 1, which contains
(1) subdomain (1A), comprising the VL domain of a monoclonal antibody capable of binding to CD123 (VLCD123) and having the amino acid sequence of SEQ ID NO: 25; and
(2) subdomain (1B), which contains the VH domain of a monoclonal antibody capable of binding to CD3 (VHCD3) and which has the amino acid sequence of SEQ ID NO: 22, wherein said subdomains 1A and 1B are separated from each other by a peptide linker having the amino acid sequence of SEK ID NO: 29;
ii. domain 2, wherein said domain 2 is a K-helix domain having the amino acid sequence SEQ ID NO: 35; and or an E-coil domain having the amino acid sequence of SEQ ID NO: 34, wherein said domain 2 is separated from said domain 1 by a peptide linker having the amino acid sequence of SEQ ID NO: 30; and wherein said domain 2 of said first and said second polypeptide chains are not both E-helix domains or both K-helix domains,
and whereby:
(a) said VL domain of said first polypeptide chain and said VH domain of said second polypeptide chain form an antigen binding domain capable of specific binding to the CD3 epitope; and
(b) said VL domain of said second polypeptide chain and said VH domain of said first polypeptide chain form an antigen binding domain capable of specific binding to the CD123 epitope.
2. Diatijelo sukladno patentnom zahtjevu 1, naznačeno time da spomenuta prvi polipeptidni lanac dodatno sadrži domenu za vezanje albumina koji ima aminokiselinsku sekvencu SEK ID BR: 36 povezan sa navedenom domenom 2 putem peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 31.2. Dibody according to claim 1, characterized in that said first polypeptide chain additionally contains an albumin binding domain having amino acid sequence SEQ ID NO: 36 connected to said domain 2 via a peptide linker having amino acid sequence SEQ ID NO: 31.
3. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da spomenuti drugi polipeptidni lanac dodatno sadrži domenu 3 koja obuhvaća CH2 i CH3 domenu imunoglobulinsku Fc domenu koja ima aminokiselinsku sekvencu SEK ID BR: 37, pri čemu je navedena domena 3 povezana s navedenom domenom 1 putem peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 33.3. Diet according to patent claim 1, characterized in that said second polypeptide chain additionally contains domain 3, which includes CH2 and CH3 domains, an immunoglobulin Fc domain having amino acid sequence SEQ ID NO: 37, wherein said domain 3 is connected to said domain 1 via of a peptide linker having the amino acid sequence SEQ ID NO: 33.
4. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da spomenuti prvi polipeptidni lanac dodatno sadrži domenu 3 koja obuhvaća CH2 i CH3 domenu imunoglobulinskog Fc domenu koja ima aminokiselinsku sekvencu SEK ID BR: 37, pri čemu je navedena domena 3 povezana s navedenom domenom 1 putem peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 33.4. Diet according to patent claim 1, characterized in that said first polypeptide chain additionally contains domain 3, which includes CH2 and CH3 domains of immunoglobulin Fc domain having amino acid sequence SEQ ID NO: 37, wherein said domain 3 is connected to said domain 1 via of a peptide linker having the amino acid sequence SEQ ID NO: 33.
5. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da spomenuti drugi polipeptidni lanac dodatno sadrži domenu 3 koja obuhvaća CH2 i CH3 domenu imunoglobulinskog Fc domene koja ima aminokiselinsku sekvencu SEK ID BR: 37, pri čemu je navedena domena 3 povezana s navedenom domenom 2 putem peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 32.5. Diet according to patent claim 1, characterized in that said second polypeptide chain additionally contains domain 3, which includes the CH2 and CH3 domains of the immunoglobulin Fc domain, which has the amino acid sequence SEQ ID NO: 37, wherein said domain 3 is connected to said domain 2 via of a peptide linker having the amino acid sequence SEQ ID NO: 32.
6. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da spomenuti prvi polipeptidni lanac dodatno sadrži domenu 3 koja sadrži CH2 i CH3 domenu imunoglobulinske Fc domene koja ima aminokiselinsku sekvencu SEK ID BR: 37, pri čemu je navedena domena 3 povezana s navedenom domenom 2 putem peptidnog poveznika koji ima aminokiselinsku sekvencu SEK ID BR: 32.6. Diet according to patent claim 1, characterized in that said first polypeptide chain additionally contains domain 3 which contains CH2 and CH3 domains of the immunoglobulin Fc domain which has the amino acid sequence SEQ ID NO: 37, wherein said domain 3 is connected to said domain 2 via of a peptide linker having the amino acid sequence SEQ ID NO: 32.
7. Dijatijelo sukladno bilo kojem od patentnih zahtjeva 3-6, naznačeno time, da spomenuto diatijelo nadalje sadrži treći polipeptidni lanac koji sadrži CH2 i CH3 domenu imunoglobulinske Fc domene koja ima aminokiselinsku sekvencu SEK ID BR: 11.7. Diabody according to any one of patent claims 3-6, characterized in that said diabody further contains a third polypeptide chain containing the CH2 and CH3 domains of the immunoglobulin Fc domain having the amino acid sequence SEQ ID NO: 11.
8. Dijatijelo sukladno bilo kojem od patentnih zahtjeva 3-7, dalje sadrži peptid koja sadrži cistein N terminalno sukladno spomenutom CH2 i CH3 domenu spomenute imunoglobulinske Fc domene, spomenuti peptid koji sadrži cistein ima aminokiselinsku sekvencu SEK ID BR: 55.8. Dietary body according to any one of claims 3-7, further comprising a peptide containing cysteine N terminally in accordance with the mentioned CH2 and CH3 domain of the mentioned immunoglobulin Fc domain, said peptide containing cysteine has the amino acid sequence SEQ ID NO: 55.
9. Dijatijelo sukladno bilo kojem od patentnih zahtjeva 1 do 8, pri čemu je spomenuta domena 2 spomenutog prvog polipeptidnog lanca domena K-zavojnice koja ima aminokiselinsku sekvencu SEK ID BR: 35 i spomenuta domena 2 spomenutog drugog polipeptidnog lanca je domena E-zavojnice koja ima aminokiselinsku sekvencu SEK ID BR: 34.9. A dibody according to any one of claims 1 to 8, wherein said domain 2 of said first polypeptide chain is a K-helix domain having the amino acid sequence SEQ ID NO: 35 and said domain 2 of said second polypeptide chain is an E-helix domain which has the amino acid sequence SEQ ID NO: 34.
10. Dijatijelo sukladno bilo kojem od patentnih zahtgeva 1 do 8, pri čemu je spomenuta domena 2 spomenutog prvog polipeptidnog lanca domena E-zavojnice koja ima aminokiselinsku sekvencu SEK ID BR: 34 i spomenuta domena 2 spomenutog drugog polipeptidnog lanca je K-spiralna domena koja ima aminokiselinsku sekvencu SEK ID BR: 35.10. The body according to any one of claims 1 to 8, wherein said domain 2 of said first polypeptide chain is an E-coil domain having the amino acid sequence SEQ ID NO: 34 and said domain 2 of said second polypeptide chain is a K-helix domain which has the amino acid sequence SEQ ID NO: 35.
11. Dijatijelo sukladno bilo kojem od prethodnih patentnih zahtjeva, naznačeno time da je dijatijelo sposobno za unakrsnu reakciju ljudskih i s proteinima CD123 i CD3 primata.11. Diabody according to any of the preceding patent claims, characterized in that the diabody is capable of cross-reacting with human and primate CD123 and CD3 proteins.
12. Dijatijelo sukladno patentnom zahtjevu 1, pri čemu:
A. prvi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: l; i
B. drugi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 3.12. Diet according to patent claim 1, wherein:
A. the first polypeptide chain contains the amino acid sequence SEQ ID NO: 1; and
B. the second polypeptide chain contains the amino acid sequence SEQ ID NO: 3.
13. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da dijatijelo dalje sadrži treći polipeptidni lanac, pri čemu:
A. prvi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 15;
B. drugi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 13; i
C. treći polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 54.13. Diet according to patent claim 1, characterized in that the diet further contains a third polypeptide chain, wherein:
A. the first polypeptide chain contains the amino acid sequence SEQ ID NO: 15;
B. the second polypeptide chain contains the amino acid sequence SEQ ID NO: 13; and
C. the third polypeptide chain contains the amino acid sequence SEQ ID NO: 54.
14. Dijatijelo sukladno patentnom zahtjevu 1, naznačeno time da dijatijelo dalje sadrži treći polipeptidni lanac, pri čemu:
A. prvi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 1;
B. drugi polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 17; i
C. treći polipeptidni lanac sadrži aminokiselinsku sekvencu SEK ID BR: 54.14. Diet according to patent claim 1, characterized in that the diet further contains a third polypeptide chain, wherein:
A. the first polypeptide chain contains the amino acid sequence SEQ ID NO: 1;
B. the second polypeptide chain contains the amino acid sequence SEQ ID NO: 17; and
C. the third polypeptide chain contains the amino acid sequence SEQ ID NO: 54.
15. Dijatijelo sukladno bilo kojem od prethodnih patentnih zahtjeva, za uporabu kao lijek.15. Dietary composition according to any of the previous patent claims, for use as medicine.
16. Dijatijelo sukladno bilo kojem od patentnih zahtjeva 1-14, za liječenje bolesti ili stanja povezanih s ili karakteriziranih ekspresijom CD123, pri čemu je navedena bolest ili stanje karcinom ili upalno stanje.16. A diet according to any one of claims 1-14, for the treatment of a disease or condition associated with or characterized by the expression of CD123, wherein said disease or condition is cancer or an inflammatory condition.
17. Dijatijelo za uporabu sukladno patentnom zahtjevu 16, naznačeno time da je spomenuta bolest ili stanje karcinom, a spomenuti karcinom je izabran iz skupine koju čine: akutna mijeloična leukemija (AML), kronična mijelogena leukemija (CML), uključujući blastičnu krizu CML-a i Abelson onkogen udružen sa CML-om (Bcr-ABL translokacija), mijelodisplastični sindrom (MDS), akutna B limfoblastna leukemija (B-ALL), kronična limfocitna leukemija (CLL), uključujući Richterov sindrom ili Richterovu transformaciju CLL, leukemija dlakastih stanica (HCL), blastična plazmacitoidna dendritiska neoplazma (BPDCN), non-Hodgkin limfom (NHL), uključujući leukemiju mantelnih stanica (MCL) i mali limfocitni limfom (SLL), Hodgkin limfom, sistemska mastocitoza i Burkitov limfom.17. Dietary composition for use according to claim 16, characterized in that said disease or condition is cancer, and said cancer is selected from the group consisting of: acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), including blast crisis of CML and Abelson oncogene associated with CML (Bcr-ABL translocation), myelodysplastic syndrome (MDS), acute B lymphoblastic leukemia (B-ALL), chronic lymphocytic leukemia (CLL), including Richter's syndrome or Richter's transformation CLL, hairy cell leukemia ( HCL), blastic plasmacytoid dendritic neoplasm (BPDCN), non-Hodgkin lymphoma (NHL), including mantle cell leukemia (MCL) and small lymphocytic lymphoma (SLL), Hodgkin lymphoma, systemic mastocytosis and Burkitt's lymphoma.
18. Dijatijelo za uporabu sukladno patentnom zahtjevu 16, naznačeno time da je spomenuta bolest ili stanje, upalno stanje, a spomenuto upalno stanje je odabrano iz skupine sastavljene od: Autoimunog lupusa (SLE), alergije, astme i reumatoidnog artritisa.18. Dietary composition for use according to claim 16, characterized in that said disease or condition is an inflammatory condition, and said inflammatory condition is selected from the group consisting of: Autoimmune Lupus (SLE), allergy, asthma and rheumatoid arthritis.
19. Farmaceutski pripravak koji sadrži dijatijelo sukladno bilo kojem od patentnih zahtjeva 1 do 14 i fiziološki prihvatljiv nosač.19. A pharmaceutical preparation containing a dibody according to any of claims 1 to 14 and a physiologically acceptable carrier.
20. Farmaceutski pripravak sukladno patentnom zahtjevu 19, za liječenje bolesti ili stanja povezanih s ili karakteriziranih ekspresijom CD123, pri čemu je navedena bolest ili stanje karcinom ili upalno stanje.20. A pharmaceutical preparation according to patent claim 19, for the treatment of diseases or conditions associated with or characterized by the expression of CD123, wherein said disease or condition is cancer or an inflammatory condition.
21. Farmaceutski pripravak za uporabu sukladno patentnom zahtjevu 20, naznačen time, da je navedena bolest ili stanje karcinom, i pri čemu je spomenuti karcinom izabran iz skupine koju čine: akutna mijeloična leukemija (AML), kronična mijelogena leukemija (CML), uključujući blastičnu krizu CML-a i Abelson onkogen udružen sa CML-om (Bcr-ABL translokacija), mijelodisplastični sindrom (MDS), akutna B limfoblastna leukemija (B-ALL), kronična limfocitna leukemija (CLL), uključujući Richterov sindrom ili Richterovu transformaciju CLL, leukemija dlakastih stanica (HCL), blastična plazmacitoidna dendritska neoplazma (BPDCN), non-Hodgkin limfom (NHL), uključujući leukemiju mantelnih stanica (MCL) i mali limfocitni limfom (SLL), Hodgkin limfom, sistemska mastocitoza i Burkitov limfom.21. Pharmaceutical preparation for use according to patent claim 20, characterized in that said disease or condition is cancer, and wherein said cancer is selected from the group consisting of: acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), including blast CML crisis and Abelson oncogene associated with CML (Bcr-ABL translocation), myelodysplastic syndrome (MDS), acute B lymphoblastic leukemia (B-ALL), chronic lymphocytic leukemia (CLL), including Richter's syndrome or Richter's transformation CLL, hairy cell leukemia (HCL), blastic plasmacytoid dendritic neoplasm (BPDCN), non-Hodgkin lymphoma (NHL), including mantle cell leukemia (MCL) and small lymphocytic lymphoma (SLL), Hodgkin lymphoma, systemic mastocytosis and Burkitt's lymphoma.
22. Farmaceutski pripravak za uporabu sukladno patentnom zahtjevu 20, naznačen time, da je navedena bolest ili stanje upalno stanje, pri čemu je spomenuto upalno stanje izabrano iz skupine koju čine: Autoimuni lupus (SLE), alergija, astma i reumatoidni artritis.22. Pharmaceutical preparation for use according to patent claim 20, characterized in that said disease or condition is an inflammatory condition, wherein said inflammatory condition is selected from the group consisting of: Autoimmune lupus (SLE), allergy, asthma and rheumatoid arthritis.