Granted Patent B2

US 12,371,486 · App. 17/236,684 · Granted Jul 29, 2025

Inventors: Mikael Maksimow (Turku, FI); Markku Jalkanen (Piispanristi, FI); Marita Vainio (Turku, FI)

Assignee: FARON PHARMACEUTICALS OY

C07K16/28A61K39/39A61K39/395A61P35/00A61P37/04C07K2317/34C07K2317/76

Abstract

Claims (101)

1. A method for treating a patient having a cancer that has CLEVER- 1 positive cells in the tumor microenvironment, the method comprising administering a humanized antibody or single chain Fv or Fab fragment capable of binding to human CLEVER-1 to the patient, wherein said humanized antibody or single chain Fv or Fab fragment comprises the following sequences of complementarity determining regions (CDRs)

i) of the heavy chain

CDR 1:

(SEQ ID NO: 7)

TSGMGIG,

CDR 2:

(SEQ ID NO: 8)

HIWWDDDKRYNPALKS,

and

CDR 3:

(SEQ ID NO: 9)

HYGYDPYYAMDY;

and

ii) of the light chain

CDR 1:

(SEQ ID NO: 10)

TASSSVSSSYLH,

CDR 2:

(SEQ ID NO: 11)

RTSNLAS,

and

CDR 3:

(SEQ ID NO: 12)

HQYHRSPPT.

2. The method according to claim 1 , wherein the humanized antibody or single chain FV or Fab fragment comprises constant regions of human IgG4 heavy chain and kappa light chain.

3. The method according to claim 1 , wherein the humanized antibody or single chain Fv or Fab fragment comprises a human IgG heavy chain variable region sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO 18 and SEQ ID NO: 20, and a human IgG light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28 and SEQ ID NO: 30.

4. The method according to claim 3 , wherein the humanized antibody or the single chain Fv or Fab fragment is capable of binding to human CLEVER-1 with a relative IC50<1.0 in comparison to the IC50 of monoclonal antibody 3-372 (DSM ACC2520 deposited at DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH on Aug. 21, 2001).

5. The method according to claim 3 , wherein the humanized antibody or the single chain Fv or Fab fragment is capable of binding to human CLEVER-1 with a relative IC50<0.8, <0.6 or <0.5 in comparison to the IC50 of monoclonal antibody 3-372.

6. The method according to claim 3 , wherein the humanized antibody or the single chain Fv or Fab fragment is capable of binding to human CLEVER-1 with a relative IC50<0.5 in comparison to the IC50 of monoclonal antibody 3-372.

7. The method according to claim 3 , wherein the combination of the human lgG heavy and light chain variable regions is selected from the group consisting of the following combinations:

SEQ ID NO: 14 and SEQ ID NO: 22;

SEQ ID NO: 16 and SEQ ID NO: 22;

SEQ ID NO: 16 and SEQ ID NO: 24;

SEQ ID NO: 16 and SEQ ID NO: 26;

SEQ ID NO: 16 and SEQ ID NO: 28;

SEQ ID NO: 16 and SEQ ID NO: 30;

SEQ ID NO: 18 and SEQ ID NO: 22;

SEQ ID NO: 18 and SEQ ID NO: 24;

SEQ ID NO: 18 and SEQ ID NO: 28;

SEQ ID NO: 18 and SEQ ID NO: 30;

SEQ ID NO: 20 and SEQ ID NO: 24; and

SEQ ID NO: 20 and SEQ ID NO: 30.

8. The method according to claim 3 , wherein the combination of human IgG heavy and light chain variable region sequence is selected from the group consisting of the following combinations: SEQ ID NO: 16 and SEQ ID NO: 30; SEQ ID NO: 18 and SEQ ID NO: 30; and SEQ ID NO: 20 and SEQ ID NO: 30.

9. The method according to claim 2 , wherein said antibody or single chain FV or Fab fragment comprises constant regions of human lgG4 heavy chain and kappa light chain with the mutations L248E and/or S241P.

10. The method according to claim 1 , wherein the method removes tumor or antigen induced immunosuppression by modulating M2 macrophages into M1 macrophages.

11. A method for reducing malignant tumor growth in an individual who has a tumor comprising CLEVER-1 positive cells in the tumor microenvironment; and/or inhibiting cancer cell transmigration and metastasis formation in a tumor comprising CLEVER-1 positive cells in the tumor microenvironment, the method comprising administering a humanized antibody or single chain Fv or Fab fragment capable of binding to human CLEVER-1 to the individual, wherein said humanized antibody or single chain Fv or Fab fragment comprises the following sequences of complementarity determining regions (CDRs)

i) of the heavy chain

CDR 1:

(SEQ ID NO: 7)

TSGMGIG,

CDR 2:

(SEQ ID NO: 8)

HIWWDDDKRYNPALKS,

and

CDR 3:

(SEQ ID NO: 9)

HYGYDPYYAMDY;

and

ii) of the light chain

CDR 1:

(SEQ ID NO: 10)

TASSSVSSSYLH,

CDR 2:

(SEQ ID NO: 11)

RTSNLAS,

and

CDR 3:

(SEQ ID NO: 12)

HQYHRSPPT.

12. The method according to claim 1 , wherein the antibody or single chain Fv or Fab fragment binds to the epitope sequences:

(SEQ ID NO: 1)

PFTVLVPSVSSFSSR,

and

(SEQ ID NO: 2)

QEITVTFNQFTK

on human CLEVER-1.

13. The method according to claim 1 , wherein the antibody or single chain Fv or Fab fragment binds to one or more epitope sequences selected from the group consisting of

(SEQ ID NO: 3)

ATQTGRVFLQ,

(SEQ ID NO: 4)

DSLRDGRLIYLF,

(SEQ ID NO: 5)

SKGRILTMANQVL,

and

(SEQ ID NO: 6)

LCVYQKPGQAFCTCR

on human CLEVER-1.

14. The method according to claim 1 , wherein the antibody or single chain Fv or Fab fragment is administered intravenously, intraarticularly, intratumorally or subcutaneously.

15. The method according to claim 11 , wherein the humanized antibody or single chain FV or Fab fragment comprises constant regions of human IgG4 heavy chain and kappa light chain.

16. The method according to claim 11 , wherein the humanized antibody or single chain Fv or Fab fragment comprises a human IgG heavy chain variable region sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO 18 and SEQ ID NO: 20, and a human IgG light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28 and SEQ ID NO: 30.

17. The method according to claim 11 , wherein the antibody or single chain Fv or Fab fragment is administered intravenously, intraarticularly, intratumorally or subcutaneously.

18. The method according to claim 1 , wherein the cancer is breast cancer.

19. The method according to claim 11 , wherein the individual has breast cancer.

20. The method according to claim 1 , wherein the cancer is leukemia.

21. The method according to claim 11 , wherein the individual has leukemia.

22. The method according to claim 1 , wherein the cancer is hepatocellular carcinoma.

23. The method according to claim 11 , wherein the individual has hepatocellular carcinoma.

24. The method according to claim 1 , wherein the cancer is bronchogenic carcinoma.

25. The method according to claim 11 , wherein the individual has bronchogenic carcinoma.

26. The method according to claim 1 , wherein the cancer is melanoma.

27. The method according to claim 11 , wherein the individual has melanoma.

Assignments (4)

Priority Claims (2)

Continuity (2)

References Cited (51)