Granted Patent B2

US 12,263,234 · App. 17/425,296 · Granted Apr 1, 2025

Inventors: Lieping Chen (Beijing, CN); Zhenguo Wen (Beijing, CN); Liqun Luo (Beijing, CN); Qianyong Liu (Beijing, CN)

Assignee: Tayu Huaxia Biotech Medical Group Co., Ltd.

A61K51/1093C07K16/2827A61K2039/505C07K2317/56C07K2317/622C07K2317/626C07K2317/92

Abstract

Claims (46)

1. An isolated anti-PD-L1 diabody comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises a first heavy chain variable region (V H-1 ) and a first light chain variable region (V L-1 ), and wherein the second polypeptide comprises a second heavy chain variable region (V H-2 ) and a second light chain variable region (V L-2 ), and wherein:

a) the V H-1 and the V H-2 each comprises an HC-CDR1 comprising the amino acid sequence of SEQ ID NO: 1, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 6, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 11; and

b) the V L-1 and the V L-2 each comprises an LC-CDR1 comprising the amino acid sequence of SEQ ID NO: 14, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 17, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 20.

2. An isolated anti-PD-L1 diabody comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises a first heavy chain variable region (V H-1 ) and a first light chain variable region (V L-1 ), and wherein the second polypeptide comprises a second heavy chain variable region (V H-2 ) and a second light chain variable region (V L-2 ), and wherein:

a) the V H-1 and the V H-2 each comprises a HC-CDR1, a HC-CDR2, and a HC-CDR3, respectively comprising the amino acid sequences of a CDR1, a CDR2, and a CDR3 within a heavy chain variable region (V H ) having the sequence set forth in any of SEQ ID NOs: 22-24; and

b) the V L-1 and the V L-2 each comprises a LC-CDR1, a LC-CDR2, and a LC-CDR3, respectively comprising the amino acid sequences of a CDR1, a CDR2, and a CDR3 within a light chain variable region (V L ) having the sequence set forth in any of SEQ ID NOs: 25-27.

3. The diabody of claim 1 , wherein the first polypeptide and the second polypeptide are linked via a covalent bond.

4. The diabody of claim 3 , wherein the covalent bond comprises a disulfide bond.

5. The diabody of claim 4 , wherein the disulfide bond is formed by a) a cysteine residue in a HC-FR2 of the first polypeptide and a cysteine residue in a LC-FR4 of the second polypeptide, and b) a cysteine residue in a LC-FR4 of the first polypeptide and a cysteine residue in a HC-FR2 of the second polypeptide.

6. The diabody of claim 1 , wherein:

a) the V H-1 in the first polypeptide and the V H-2 in the second polypeptide comprises a G44C mutation or a Q105C mutation according to the Kabat numbering system; and/or

b) the V L-1 in the first polypeptide and the V L-2 in the second polypeptide comprises a Q100C mutation or an A43C mutation according to the Kabat numbering system.

7. The diabody of claim 1 , wherein the V H-1 and the V L-1 of the first polypeptide and the V H-2 and the V L-2 of the second polypeptide are fused to each other via a peptide linker.

8. The diabody of claim 1 , wherein the V H-1 and V H-2 comprises the amino acid sequence of any one of SEQ ID NOs: 22-24, and the V L-1 and V L-2 comprises the amino acid sequence of any one of SEQ ID NOs: 25-27.

9. The diabody of claim 1 , wherein the first polypeptide and the second polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 36-43.

10. A pharmaceutical composition comprising the diabody of claim 1 , and a pharmaceutical acceptable carrier.

11. A polynucleotide encoding the diabody of claim 1 .

12. A nucleic acid construct, comprising the polynucleotide of claim 11 .

13. A vector comprising the nucleic acid construct of claim 12 .

14. An isolated host cell comprising the polynucleotide according to claim 11 .

15. A culture medium comprising the diabody of claim 1 .

16. A method of producing an anti-PD-L1 diabody, comprising:

a) culturing the isolated host cell of claim 14 under conditions effective to express the diabody; and

b) obtaining the expressed diabody from the host cell.

17. A method of determining the distribution of PD-L1 in an individual, comprising:

a) administering to the individual an imaging agent comprising the diabody of claim 1 labeled with a radionuclide; and

b) imaging the imaging agent in the individual with a non-invasive imaging technique.

18. A method of diagnosing an individual having a PD-L1 associated disease or condition, comprising:

determining the distribution of PD-L1 in the individual using the method of claim 17 ,

wherein the individual is positive for PD-L1 if signal of the imaging agent is detected at a tissue of interest, or the individual is negative for PD-L1 if signal of the imaging agent is not detected at a tissue of interest.

19. A method of treating an individual having a PD-L1 associated disease or condition, comprising:

a) diagnosing the individual using the method of claim 18 ; and

b) administering to the individual an effective amount of a therapeutic agent targeting PD-L1, if the individual is positive for PD-L1.

20. A method of treating an individual having a disease or condition, comprising administering to the individual an effective amount of the diabody of claim 1 .

21. An imaging agent comprising the diabody of claim 1 labeled with a radionuclide.

22. A method of preparing an imaging agent targeting PD-L1, comprising:

a) conjugating a chelating compound to the diabody of claim 1 to provide an anti-PD-L1 diabody conjugate; and

b) contacting a radionuclide with the anti-PD-L1 diabody conjugate, thereby providing the imaging agent.

23. A method of determining the distribution of PD-L1 in an individual, comprising:

(a) administering to the individual an effective amount of a diabody agent comprising the diabody of claim 1 and a first conjugation moiety;

(b) subsequently administering to the individual an effective amount of a radionuclide compound comprising a radionuclide and a second conjugation moiety, wherein the first conjugation moiety and the second conjugation moiety is conjugated to each other in vivo to provide an imaging agent; and

(c) imaging the imaging agent in the individual with a non-invasive imaging technique.

24. The diabody of claim 8 , wherein:

a) the V H-1 and V H-2 comprises the amino acid sequence of SEQ ID NO: 22, and the V L-1 and V L-2 comprises the amino acid sequence of SEQ ID NO: 25;

b) the V H-1 and V H-2 comprises the amino acid sequence of SEQ ID NO: 23, and the V L-1 and V L-2 comprises the amino acid sequence of SEQ ID NO: 26; or

c) the V H-1 and V H-2 comprises the amino acid sequence of SEQ ID NO: 24, and the V L-1 and V L-2 comprises the amino acid sequence of SEQ ID NO: 27.

Assignments (1)

Continuity (1)

References Cited (322)