Anti-cancer combination therapy

文档序号：1835522 发布日期：2021-11-12 浏览：22次中文

阅读说明：本技术 抗癌组合疗法 (Anti-cancer combination therapy ) 是由 V·津萨拉 B·德罗比特·汉德尔 M·J·鲍尔于 2020-03-26 设计创作，主要内容包括：本发明描述抗癌疗法,其包括使用LRP5拮抗剂与抗PD1抗体(各如本申请描述)组合。(The present invention describes anti-cancer therapies comprising the use of LRP5 antagonists in combination with anti-PD 1 antibodies (each as described herein).)

1. An LRP5 antagonist for use in a method for treating and/or preventing a hyperproliferative disease, preferably cancer, wherein the method comprises administering to a patient in need thereof the LRP 5-antagonist in combination with a PD-1 antibody,

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

(i) an anti-PD 1 antibody comprising heavy chain CDRs comprising the amino acid sequences of SEQ ID NO:1(HCDR1), SEQ ID NO:2(HCDR2) and SEQ ID NO:3(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO:4(LCDR1), SEQ ID NO:5(LCDR2) and SEQ ID NO:6(LCDR 3);

(ii) an anti-PD 1 antibody comprising heavy chain CDRs comprising the amino acid sequences of SEQ ID NO 7(HCDR1), SEQ ID NO 8(HCDR2) and SEQ ID NO 9(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO 10(LCDR1), SEQ ID NO 11(LCDR2) and SEQ ID NO 12(LCDR 3); and

(iii) an anti-PD 1 antibody comprising heavy chain CDRs comprising the amino acid sequences of SEQ ID NO 13(HCDR1), SEQ ID NO 14(HCDR2) and SEQ ID NO 15(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO 16(LCDR1), SEQ ID NO 17(LCDR2) and SEQ ID NO 18(LCDR 3).

2. A method of treating and/or preventing a hyperproliferative disease, preferably cancer, comprising administering to a patient in need thereof a therapeutically effective amount of an LRP5 antagonist and a therapeutically effective amount of a PD-1-antibody;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

3. A PD-1 antibody for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein said method comprises administering to a patient in need thereof the PD-1 antibody in combination with an LRP 5-antagonist:

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

Use of an LRP5 antagonist for the preparation of a pharmaceutical composition for use in a method for the treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the LRP 5-antagonist is used in combination with a PD-1 antibody,

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

Use of a PD-1 antibody for the preparation of a pharmaceutical composition for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the PD-1-antibody is used in combination with an LRP5 antagonist;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

6. A pharmaceutical composition, comprising:

LRP5 antagonists;

a PD-1 antibody; and

optionally, one or more pharmaceutically acceptable carriers, excipients and/or vehicles;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

7. The pharmaceutical composition according to claim 6 for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer.

8. A kit comprising, in one or more containers:

a first pharmaceutical composition or dosage form comprising an LRP5 antagonist and optionally one or more pharmaceutically acceptable carriers, excipients, and/or vehicles;

a second pharmaceutical composition or dosage form comprising the PD-1 antibody and optionally one or more pharmaceutically acceptable carriers, excipients, and/or vehicles;

and optionally a package insert comprising printed instructions;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising:

(a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising:

(a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and wherein the PD-1 antibody is selected from the group consisting of:

9. The kit according to claim 8 for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer.

10. An LRP5 antagonist for use according to claim 1, a method of treatment according to claim 2, a PD-1 antibody for use according to claim 3, a use according to claim 4 or 5, a pharmaceutical composition according to claim 6 or 7, or a kit according to claim 8 or 9, wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO. 50 and a second ISVD comprising the sequence of SEQ ID NO. 61;

(ii) a polypeptide comprising a first ISVG comprising the amino acid sequence of SEQ ID No. 51 and a second ISVD comprising the sequence of SEQ ID No. 52;

(iii) a polypeptide comprising a first ISVD comprising the sequence of SEQ ID NO:62 and a second ISVD comprising the sequence of SEQ ID NO: 53;

(iv) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO. 50 and a second ISVD comprising the sequence of SEQ ID NO. 53;

(v) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO:62 and a second ISVD comprising the sequence of SEQ ID NO: 61; and

(vi) a polypeptide comprising a first ISVD comprising the amino acid sequence of SEQ ID NO. 51 and a second ISVD comprising the sequence of SEQ ID NO. 53;

preferably wherein the LRP5 antagonist further comprises an Alb11 domain comprising the amino acid sequence of SEQ ID NO: 60.

11. An LRP5 antagonist for use according to claim 1, a method of treatment according to claim 2, a PD-1 antibody for use according to claim 3, a use according to claim 4 or 5, a pharmaceutical composition according to claim 6 or 7, or a kit according to claim 8 or 9, wherein the LRP5 antagonist comprises a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO 57, SEQ ID NO 58, and SEQ ID NO 59.

12. The LRP5 antagonist for use according to claim 1, 10 or 11, the method of treatment according to claim 2, 10 or 11, the PD-1 antibody for use according to claim 3, 10 or 11, the use according to claim 4, 5, 10 or 11, the pharmaceutical composition according to claim 6, 7, 10 or 11, or the kit according to claim 8, 9, 10 or 11, wherein the anti-PD 1 antibody is selected from the group consisting of:

(i) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 20;

(ii) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 22;

(iii) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 24;

(iv) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 26; and

(v) an antibody having a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 28.

13. An LRP5 antagonist for use according to claim 1, 10 or 11, a method of treatment according to claim 2, 10 or 11, a PD-1 antibody for use according to claim 3, 10 or 11, a use according to claim 4, 5, 10 or 11, a pharmaceutical composition according to claim 6, 7, 10 or 11, or a kit according to claim 8, 9, 10 or 11, wherein the PD-1 antibody is selected from the group consisting of:

(i) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO. 29 and a light chain comprising the amino acid sequence of SEQ ID NO. 30;

(ii) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO. 31 and a light chain comprising the amino acid sequence of SEQ ID NO. 32;

(iii) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO. 33 and a light chain comprising the amino acid sequence of SEQ ID NO. 34;

(iv) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO 35 and a light chain comprising the amino acid sequence of SEQ ID NO 36; and

(v) an antibody having a heavy chain comprising the amino acid sequence of SEQ ID NO. 37 and a light chain comprising the amino acid sequence of SEQ ID NO. 38.

14. The LRP5 antagonist for use according to claim 1, 10, 11, 12 or 13, the method of treatment according to claim 2, 10, 11, 12 or 13, the PD-1 antibody for use according to claim 3, 10, 11, 12 or 13, the use according to claim 4, 5, 10, 11, 12 or 13, or the kit for use according to claim 9, 10, 11, 12 or 13, wherein the PD-1 antibody and the LRP5 antagonist are administered simultaneously, concurrently, sequentially, alternately or separately.

15. The LRP5 antagonist for use according to claim 1, 10, 11, 12 or 13, the method of treatment according to claim 2, 10, 11, 12 or 13, the PD-1 antibody for use according to claim 3, 10, 11, 12 or 13, the use according to claim 4, 5, 10, 11, 12 or 13, or the kit for use according to claim 9, 10, 11, 12 or 13, wherein the LRP5 antagonist and the PD-1 antibody are administered according to the following treatment regimen:

(i) a first treatment period, wherein the LRP5 antagonist and the PD-1 antibody are administered simultaneously or concurrently, preferably every three or four weeks; and

(ii) a second treatment period, wherein only the PD-1 antibody is administered and the LRP5 antagonist is not administered, preferably wherein the PD-1 antibody is administered every three or four weeks.

16. The LRP5 antagonist, the method of treatment, the PD-1 antibody for use, the use or the kit for use according to claim 15, wherein the first treatment period is 3 or 6 weeks when the LRP5 antagonist and PD-1 antibody are administered every three weeks, or 4 or 8 weeks when the LRP5 antagonist and PD-1 antibody are administered every four weeks.

17. The LRP5 antagonist, the method of treatment, the PD-1 antibody used, the use, the pharmaceutical composition used, or the kit of use, for use according to claim 14, 15, or 16, wherein the administration is intravenous administration.

18. The LRP5 antagonist for use according to any one of claims 1, 10 to 17, the method of treatment according to any one of claims 2, 10 to 17, the PD-1 antibody for use according to any one of claims 3, 10 to 17, the use according to any one of claims 4, 5, 10 to 17, the pharmaceutical composition for use according to any one of claims 7, 10 to 17 or the kit for use according to any one of claims 9 to 17, wherein the hyperproliferative disease to be treated is selected from the group of cancers consisting of: gastrointestinal cancer, melanoma tumor, bladder cancer, and lung cancer (e.g., NSCLC).

19. An LRP5 antagonist, a method of treatment, a PD-1 antibody for use, a pharmaceutical composition for use, or a kit for use according to claim 18, wherein the cancer of the gastrointestinal tract is cancer of the esophagus (e.g. gastroesophageal junction cancer), stomach (stomach) cancer, hepatocellular cancer, biliary tract cancer (e.g. cholangiocarcinoma), gall bladder cancer, pancreatic cancer, or colorectal cancer (CRC).

20. The LRP5 antagonist, the method of treatment, the PD-1 antibody used, the use, the pharmaceutical composition used or the kit of use for use according to claim 18 or 19, wherein the cancer is an anti-immunotherapeutic tumour.

21. The LRP5 antagonist for use according to any one of claims 1, 10 to 17, the method of treatment according to any one of claims 2, 10 to 17, the PD-1 antibody for use according to any one of claims 3, 10 to 17, the use according to any one of claims 4, 5, 10 to 17, the pharmaceutical composition for use according to any one of claims 7, 10 to 17 or the kit for use according to any one of claims 9 to 17, wherein the hyperproliferative disease to be treated is an anti-immunotherapeutic solid tumor.

Technical Field

The present invention relates to combination therapies for the treatment of cancer and to compounds for use in such combination therapies. The compounds for use in combination are LRP5 antagonists and PD-1 antagonists.

Background

Activation of the Wnt signaling pathway requires binding of extracellular Wnt ligand to the frizzled receptor and to the co-receptor LRP5 (accession No.: UniProtKB-O75197/LRP5_ HUMAN) or its closely related homologous LRP6 (accession No.: UniProtKB-O75581/LRP6_ HUMAN). There are 19 Wnt proteins and 10 frizzled receptors in mammalian cells. In the absence of Wnt ligands, cytoplasmic β -catenin is phosphorylated by a protein complex consisting of the scaffold protein Axin (Axin) and APC and the kinases GSK3 β and CK1 a. Subsequent recognition of ubiquitin ligase β -TrcP results in ubiquitin-mediated degradation of β -catenin. In the presence of Wnt ligands, Wnt binding to frizzled and LRP5 results in recruitment of cytoplasmic effector protein Dvl and phosphorylation of the LRP5 cytoplasmic tail, which provides a docking site for axin. LRP5 isolates axin resulting in inactivation of the axin-APC-GSK 3 β complex and thus stabilization and aggregation of intracellular β -catenin. Thus, the cytoplasmic concentration of β -catenin increases, and β -catenin migrates to the nucleus and complexes with members of the T-cytokine (TCF)/lymphoenhancer binding factor (LEF) family of transcription factors. Basal transcription machinery and transcriptional co-activators, including cAMP response component binding protein (CREB) -binding protein (CBP) or its homolog p300, are then recruited, resulting in the expression of various target genes, including axin 2, cyclin D1, Naked1, Notum, and c-Myc.

Additional levels of ligand-dependent Wnt pathway regulation are mediated by The E3 ligase RNF43 and its closely related homologue ZNRF3, and by secreted R-spinal protein proteins (de Lau et al, "The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength h". Genes Dev.2014; 28(4): 305-16). RNF43 mediates ubiquitination of the frizzled/LRP 5 receptor complex at the cell surface, causing its degradation, thereby inhibiting ligand-dependent Wnt pathway activity. The activity of RNF43 is counteracted by members of the R-vertebrate family (R-vertebrate 1 to 4 ligands). When present, R-spondyloprotein ligands remove RNF43 from the cell surface, allowing aggregation of the frizzled/LRP 5 complex and enhancement of Wnt signaling in the presence of Wnt ligands.

Hyperactivation of Wnt signaling is involved in the pathogenesis of various (although not all) types of cancer in at least two different ways: in some cancer types, frequent mutations in downstream signaling molecules contribute to the structurally activated Wnt pathway (e.g., APC mutations in colorectal cancer; β -catenin activating mutations in hepatocellular carcinoma), while in other types of cancer, e.g., Triple Negative Breast Cancer (TNBC), non-small cell lung cancer (NSCLC), pancreatic cancer, and in the endometrium and colorectal cancer (CRC), Wnt signaling activation is driven by a ligand-dependent mechanism (i.e., by autocrine/paracrine Wnt activation), as detected by β -catenin intracellular aggregation. Ligand-dependent Wnt activation is mediated in NSCLC, TNBC, and Pancreatic Cancer by a variety of mechanisms, including increased expression of Wnt ligand and/or LRP5 receptor, or silencing of the LRP5 negative regulator DKK1 (TNBC: Khramtsov et al, "Wnt/beta-catenin pathway activity expressed in basic-lipid complexes and precursors pore outer genome". Am J Pathol.2010; 176(6): 2911-20; NSCLC: Nakashima et al, "Wnt 1 expression with structural expression and a pore expression in non-cell lung tissues". 2008. "(19) (1): 203-9; functional expression: Zrecognition et al," Cancer expression in n-cell tissue reaction 20115: "Cancer expression". In particular, published data has shown that in healthy tissues (e.g., breast and lung epithelium), β -catenin is located only in the serosa. In contrast, most TNBC, NSCLC, and pancreatic cancer major clinical specimens show intracellular aggregation of β -catenin (i.e., in the cytoplasm/nucleus; a biomarker of Wnt signaling activation) due to aberrant Wnt signaling.

Because LRP5 acts as a gatekeeper for ligand-dependent Wnt signaling activation, it can be considered as a target to achieve complete blockade of pathways mediated by all 19 Wnt ligands and 10 frizzled receptors.

An alternative to the above described methods of directly targeting cancer/cancer cells is cancer immunotherapy. Cancer immunotherapy is a branch of oncology, where the immune system is used to treat cancer, in sharp contrast to the existing common treatment methods in which tumors are directly excised or treated. This therapeutic concept is based on the recognition of many proteins on the surface of T cells that act to suppress the immune function of these cells. An example of such a protein is PD-1 (programmed cell death protein 1).

PD-1 is a cell surface receptor protein expressed on T cells. PD-1 has two ligands, PD-L1 and PD-L2, which interact with the cell surface receptor. Upon binding of the ligand, PD-1 induces an intracellular signal that negatively regulates the T cell response. Thus, in general, the protein functions as an inhibitor of "immune checkpoints", i.e. it functions to regulate the activity of cells in the immune system in order to regulate and limit autoimmune diseases. It has recently been appreciated that many cancers can be kept away from the immune system by modifying "immune checkpoint" inhibitors and thus avoid detection.

Thus, it has also been shown that antagonistic PD-1 antibody molecules such as e.g. nivolumab (nivolumab) and pembrolizumab (pembrolizumab) can be used to stimulate the immune system and thus treat cancer in a range of different cancer situations.

Despite the above described advances in cancer treatment, there remains a need for novel treatment concepts directed to cancer treatment that exhibit advantages over standard therapies. These advantages may include in vivo utility (e.g., improved clinical response, prolongation of response, increase in response rate, duration of response, rate of disease stabilization, duration of stabilization, time to disease progression, progression-free survival (PFS) and/or Overall Survival (OS), subsequent emergence of resistance, and the like), safe and well-tolerated administration and reduced frequency and severity of adverse events. In particular, additional treatment options are needed for patients with cancer, such as, for example, lung cancer (e.g., NSCLC), melanoma, bladder cancer, and gastrointestinal cancer.

It is therefore an object of the present invention to provide novel treatments for cancer that are superior to treatments/methods currently used and/or known in the art.

Disclosure of Invention

The present invention provides methods of treating a patient having a hyperproliferative disease with an LRP5 antagonist (which term is used interchangeably herein with the terms "polypeptide that specifically binds to LRP 5" or "polypeptide that specifically binds to LRP 5"), together with an antibody specific for programmed cell death protein 1(PD-1) (which term is used interchangeably herein with the terms "anti-PD-1 antibody", "PD-1 antibody", or "PD-1 antagonist"), thereby antagonizing the PD-1 signaling pathway. Accordingly, the invention provides combination therapies comprising an LRP5 antagonist and an anti-PD-1 antibody.

In one aspect, the invention provides an LRP5 antagonist for use in a method of treating and/or preventing a hyperproliferative disease, preferably cancer, comprising administering to a patient in need thereof the LRP5 antagonist in combination with a PD-1 antibody,

the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and the PD-1 antibody is selected from the group consisting of:

In another aspect, the invention provides a method of treating and/or preventing a hyperproliferative disease, preferably cancer, comprising administering to a patient in need thereof a therapeutically effective amount of an LRP5 antagonist and a therapeutically effective amount of a PD-1 antibody,

the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

and the PD-1 antibody is selected from the group consisting of:

In another aspect, the invention provides a PD-1 antibody for use in a method of treating and/or preventing a hyperproliferative disease, preferably cancer, comprising administering to a patient in need thereof a PD-1 antibody in combination with an LRP5 antagonist,

the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

and the PD-1 antibody is selected from the group consisting of:

In a further aspect, the invention provides the use of an LRP5 antagonist for the preparation of a pharmaceutical composition for use in a method for the treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the LRP 5-antagonist is to be used in combination with a PD-1 antibody,

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

wherein the PD-1 antibody is selected from the group consisting of:

In another aspect, the invention provides the use of a PD-1 antibody for the manufacture of a pharmaceutical composition for use in a method for the treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the PD-1-antibody is to be used in combination with an LRP5 antagonist;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

wherein the PD-1 antibody is selected from the group consisting of:

In another aspect, the present invention provides a pharmaceutical composition comprising:

an LRP5 antagonist;

PD-1 antibody; and

optionally, one or more pharmaceutically acceptable carriers, excipients and/or vehicles;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and (b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and (b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and (b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

wherein the PD-1 antibody is selected from the group consisting of:

In some embodiments, the pharmaceutical composition is used in a method of treating and/or preventing a hyperproliferative disease, preferably cancer.

In another aspect, the present invention provides a kit comprising, in one or more containers:

a first pharmaceutical composition or dosage form comprising an LRP5 antagonist and optionally one or more pharmaceutically acceptable carriers, excipients, and/or vehicles;

a second pharmaceutical composition or dosage form comprising the PD-1 antibody and optionally one or more pharmaceutically acceptable carriers, excipients and/or vehicles;

and optionally a package insert comprising printed instructions;

wherein the LRP5 antagonist is selected from the group consisting of:

(i) a polypeptide comprising: (a) a first Immunoglobulin Single Variable Domain (ISVD) comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(ii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

(iii) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO: 48); and

(iv) a polypeptide comprising: (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

(b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)；

wherein the PD-1 antibody is selected from the group consisting of:

In some embodiments, the kit according to the invention is used in a method for the treatment and/or prevention of a hyperproliferative disease, preferably cancer.

In a preferred embodiment of the invention, the LRP5 antagonist is selected from the group consisting of:

(i) a first ISVD comprising the amino acid sequence of SEQ ID NO. 50, and a second ISVD comprising the sequence of SEQ ID NO. 61;

(ii) a first ISVG comprising the amino acid sequence of SEQ ID No. 51, and a second ISVD comprising the sequence of SEQ ID No. 52;

(iii) a first ISVD comprising the sequence of SEQ ID NO:62, and a second ISVD comprising the sequence of SEQ ID NO: 53;

(iv) a first ISVD comprising the amino acid sequence of SEQ ID NO. 50, and a second ISVD comprising the sequence of SEQ ID NO. 53;

(v) a first ISVD comprising the amino acid sequence of SEQ ID NO:62, and a second ISVD comprising the sequence of SEQ ID NO: 61; and

(vi) a first ISVD comprising the amino acid sequence of SEQ ID NO. 51, and a second ISVD comprising the sequence of SEQ ID NO. 53;

preferably, the LRP5 antagonist further includes an AIb11 domain comprising the amino acid sequence of SEQ ID NO 60.

In particularly preferred embodiments, the LRP5 antagonist includes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO 57, SEQ ID NO 58, and SEQ ID NO 59.

In a preferred embodiment of the invention, the anti-PD 1 antibody is selected from the group consisting of:

(i) an antibody comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 20;

(ii) an antibody comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 22;

(iii) an antibody comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 24;

(iv) an antibody comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 26; and

(v) an antibody comprising a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 28.

In a particularly preferred embodiment of the invention, the PD-1 antibody is selected from the group consisting of:

(i) an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO. 29 and a light chain comprising the amino acid sequence of SEQ ID NO. 30;

(ii) an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO. 31 and a light chain comprising the amino acid sequence of SEQ ID NO. 32;

(iii) an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO. 33 and a light chain comprising the amino acid sequence of SEQ ID NO. 34;

(iv) an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO 35 and a light chain comprising the amino acid sequence of SEQ ID NO 36; and

(v) an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO. 37 and a light chain comprising the amino acid sequence of SEQ ID NO. 38.

In some embodiments of the invention, the PD-1 antibody will be administered simultaneously, concurrently, sequentially, alternately, or separately with the LRP5 antagonist.

In preferred embodiments, the LRP5 antagonist and the PD-1 antibody will be administered according to the following treatment regimens:

(i) a first treatment cycle wherein the LRP5 antagonist and the PD-1 antibody are to be administered simultaneously or concurrently, preferably every three or four weeks; and

(ii) a second treatment cycle wherein only the PD-1 antibody and the LRP5 antagonist are to be administered, preferably wherein the PD-1 antibody is to be administered every three or four weeks.

In a preferred embodiment of the invention, the hyperproliferative disease to be treated is a cancer selected from the group consisting of: gastrointestinal cancer, melanoma tumor, bladder cancer, and lung cancer (e.g., NSCLC), even more preferably the cancer is an anti-immunotherapeutic gastrointestinal cancer (including, but not limited to, esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular cancer, biliary tract cancer (e.g., cholangiocarcinoma), gallbladder cancer, pancreatic cancer, or colorectal cancer (CRC)), an anti-immunotherapeutic melanoma, an anti-immunotherapeutic bladder cancer, or an anti-immunotherapeutic lung cancer.

In an alternative preferred embodiment of the invention, the hyperproliferative disease to be treated is an anti-immunotherapeutic solid tumor.

Brief description of the drawings

Fig. 1A to 1H: the anti-tumor activity of an exemplary LRP5 antagonist as a single agent and in combination with an exemplary PD-1 antibody in a subcutaneous isogenic mouse model derived from the breast cancer cell line EMT6 in Balb/c mice is shown. FIG. 1A: in isotype-matched antibodies; 1B: an LRP5 antagonist; 1C: with a PD-1 antibody; and 1D: measurement of tumor volume after the indicated days of treatment with LRP5 antagonist + PD-1 antibody. FIG. 1E: in isotype-matched antibodies; 1F: an LRP5 antagonist; 1G: with a PD-1 antibody; and 1H: measurement of tumor contractile response after the indicated days of treatment with LRP5 antagonist + PD-1 antibody. The numbers indicated by a-indicate the number of mice in the total study mice in which a response was observed, i.e. in which the ratio between the tumor volume at the end of the treatment and the tumor volume at the beginning of the treatment was below 1 (i.e. indicating shrinkage of the tumor).

Detailed Description

Definition of

The above and other aspects and embodiments of the invention will become apparent from the further description of the present application, in which:

unless otherwise indicated or defined herein, all terms used have their ordinary meanings in the art, which will be apparent to those skilled in the art. In case of conflict, the present patent specification, including definitions, will control. For example, reference is made to standard manuals, such as Sambrook et al, "Molecular Cloning: A Laboratory Manual" (2 nd edition), volumes 1 to 3, Cold Spring Harbor Laboratory Press (1989); lewis, "Genes IV", Oxford University Press, New York, (1990), and Roitt et al, "Immunology" (2 nd edition), Gower Medical Publishing, London, New York (1989), and for the general background of the present application. Moreover, unless otherwise indicated herein, all methods, steps, techniques and operations not specifically described in detail can be performed and performed in a manner known per se, as would be apparent to one of ordinary skill in the art. For example, reference is again made to standard manuals, to the prior art mentioned above and to other references cited in the present application.

The term "antibody" includes antibodies, antibody fragments, antibody-like molecules, and conjugates that bind to any of the above. Antibodies include, but are not limited to, polyclonal or monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, monospecific antibodies, bispecific antibodies, or multispecific antibodies. The term "antibody" shall encompass intact immunoglobulins as they are produced by lymphocytes and are present, for example, in blood serum, monoclonal antibodies secreted by hybridoma cell lines, polypeptides produced by recombinant expression in host cells which have the binding specificity of an immunoglobulin or monoclonal antibody, and molecules which have been derived from these immunoglobulins, monoclonal antibodies or polypeptides by further processing while retaining their binding specificity. In particular, the term "antibody" includes intact immunoglobulins comprising two heavy chains and two light chains. In another embodiment, the term encompasses fragments of immunoglobulins, such as Fab fragments. In another embodiment, the term "antibody" includes polypeptides having one or more variable domains derived from an immunoglobulin, such as single chain antibodies (scFv), single domain antibodies, and the like.

"human antibodies" are antibodies having amino acid sequences corresponding to those of antibodies produced by human cells or derived from non-human sources using human antibody libraries or other human antibody coding sequences. This definition of human antibodies specifically excludes humanized antibodies comprising non-human antigen binding residues.

As used herein, the term "recombinant human antibody" is intended to include all human antibodies that are prepared, expressed, created or isolated by recombinant means, such as antibodies isolated from a host cell (e.g., NS0 or CHO cells) or from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes or antibodies expressed using a recombinant expression vector transfected into a host cell. These recombinant human antibodies have variable and constant regions in rearranged form. Recombinant human antibodies according to the invention have undergone high frequency mutagenesis of endosomal cells in vivo. Thus, the amino acid sequences of the VH and VL regions of the recombinant antibodies, while derived from and related to human germline VH and VL sequences, may not be sequences naturally occurring in vivo in the human antibody germline repertoire.

"humanized" antibodies refer to chimeric antibodies comprising amino acid residues from the non-human Highly Variable Region (HVR) and amino acid residues from the human Framework Region (FR). In certain embodiments, a humanized antibody will comprise substantially all of at least one and typically two variable domains, wherein all or substantially all (e.g., Complementarity Determining Regions (CDRs)) of these HVRs correspond to those of a non-human antibody, and all or substantially the entire Framework Regions (FRs) correspond to those of a human antibody. The humanized antibody optionally may comprise at least a portion of an antibody constant region derived from a human antibody. "humanized forms" of antibodies (e.g., non-human antibodies) refer to antibodies that have undergone humanization.

The expression "variable domain" or "variable region" or Fv as used herein denotes each of the light and heavy chain pairs, which is directly involved in the binding of an antibody to an antigen. The variable domain of the light chain is abbreviated "VL" and the variable domain of the heavy chain is abbreviated "VH". The variable light and heavy chain domains have the same general structure and each domain comprises four Framework (FR) regions whose sequences are widely conserved, connected by three HVRs (or CDRs). The framework regions adopt a beta sheet conformation and the CDRs can form loops connecting the beta sheet structure. The CDRs in each chain are held in their three-dimensional structure by the framework regions and form together with the CDRs from the other chains the antigen binding site. The heavy and light chain CDR regions of the antibody play a particularly important role in the binding specificity/affinity of the antibody according to the invention and thus provide a further object of the invention.

In the context of the present invention, references to CDRs associated with an antibody (e.g., the PD1 antibody) are based on the definition of Chothia (Chothia and Lesk, J.mol.biol.1987,196:901-917), along with the definition of Kabat (E.A.Kabat, T.T.Wu, H.Bilofsky, M.Reid-Miller and H.Perry, Sequence of Proteins of Immunological Interest, National Institutes of Health, Bethesda (1983)).

Unless otherwise indicated herein, the terms "immunoglobulin" and "immunoglobulin sequence" are used as general terms to include full-size antibodies, individual chains and all portions thereof, domains or fragments thereof (including, but not limited to, antigen-binding domains or fragments such as VHH domains or VH/VL domains, respectively), whether or not used herein to refer to heavy chain antibodies or to conventional 4 chain antibodies. In addition, unless the context requires more limited interpretation, the term "sequence" as used herein (e.g. in terms such as "immunoglobulin sequence", "antibody sequence", "(single) variable domain sequence", "VHH sequence" or "protein sequence") should generally be understood to include related amino acid sequences and nucleic acid sequences or nucleotide sequences encoding the same.

The term "domain" (of a polypeptide or protein) as used herein refers to a folded protein structure that has the ability to retain its tertiary structure independent of the remainder of the protein. In general, domains are responsible for discrete functional properties of a protein, and in many cases may be added, removed, or transferred to other proteins without loss of the function of the remainder of the protein and/or domain.

The term "immunoglobulin domain" as used herein refers to a globular region of an antibody chain (e.g., a chain of a conventional 4-chain antibody or heavy chain antibody), or to a polypeptide consisting essentially of such a globular region. The immunoglobulin domain is characterized in that it retains the immunoglobulin fold properties of the antibody molecule, and is composed of a 2-layer sandwich of about 7 antiparallel beta chains arranged in two beta sheets, optionally stabilized by conserved disulfide bonds.

The term "immunoglobulin variable domain" as used herein means the "framework region 1" or "FR 1", respectively, substantially by reference in the art and in the present application; referred to as "framework region 2" or "FR 2"; referred to as "framework region 3" or "FR 3"; and four "framework regions" designated "framework region 4" or "FR 4"; these framework regions are referred to by reference in the art and herein as "complementarity determining region 1" or "CDR 1," respectively; referred to as "complementarity determining region 2" or "CDR 2"; and three "complementarity determining regions" or "CDR" discontinuities referred to as "complementarity determining region 3" or "CDR 3". Thus, the general structure or sequence of an immunoglobulin variable domain can be indicated as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR 4. It is the immunoglobulin variable domain that confers specificity to an antibody for an antigen by carrying an antigen binding site.

The term "immunoglobulin single variable domain" (or ISVD), as used herein, means an immunoglobulin variable domain that can specifically bind to an epitope of an antigen without pairing with additional variable immunoglobulin domains. An example of an ISVD within the meaning of the present invention is a "domain antibody", e.g. an ISVD VH and VL (VH domain and VL domain). Another important example of an ISVD, as defined below, is a "VHH domain" (or only a "VHH") from a camelid.

In view of the above definitions, the antigen binding domain or Fab fragment, F (ab')2 fragment, Fv fragment (e.g. disulfide-linked Fv or scFv fragment), of a conventional 4 chain antibody (e.g. IgG, IgM, IgA, IgD or IgE molecule; known in the art), or of a diabody derived from such a conventional 4 chain antibody (all known in the art) should generally not be considered an ISVD, since in these cases the individual epitopes bound to an antigen generally do not occur from one (single) immunoglobulin domain but from a pair of (associated) immunoglobulin domains (e.g. light and heavy chain variable domains), i.e. from a VH-VL pair of immunoglobulin domains that bind together to the epitopes of the individual antigen.

"VHH domain", also called VHH, V_HH domains, VHH antibody fragments and VHH antibodies have been described initially as "heavy chain antibodies" (i.e., "light chain-deficient antibodies"; Hamers-Casterman C, Atathouch T, Muydermans S, Robinson G, Ab, and Ab, to be, e.g., antibodies,Hamers C, Songa EB, Bendahman N, Hamers R. "Natural cure antibodies void of light chains"; antigen binding immunoglobulin (variable) domains of Nature363,446-448 (1993)). The term "VHH domain" has been selected to distinguish these variable domains from the heavy chain variable domains (which are referred to herein as "V" s) present in conventional 4 chain antibodies_HDomain "or" VH domain ") and the light chain variable domain present in conventional 4 chain antibodies (which is referred to herein as" V "in this application)_LDomain "or" VL domain "). The VHH domain may specifically bind to an epitope without an additional antigen-binding domain (relative to the VH or VL domain in conventional 4 chain antibodies, in which case the epitope is recognized by the VL domain together with the VH domain). The VHH domain is a small, robust and efficient antigen recognition unit formed by a single immunoglobulin domain.

In the context of the present invention, the terms VHH domain, VHH, V_HH-domains, VHH antibody fragments, VHH antibodies and' nanobodies'and' nano antibodyDomains "(a" nanobody "being a trademark of the company Ablynx n.v.; Ghent; Belgium) are used interchangeably and are as defined, for example, in WO2009/109635, fig.1, denoting ISVD (having the structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 and specifically binding to an epitope without the presence of a second immunoglobulin variable domain) and which can also be distinguished from VH domains by so-called" marker residues ".

For example, as shown in FIG. 2 of Riechmann and Muydermans, J.Immunol.methods 231,25-38(1999), as applied to a VHH domain from a camelid, the amino acid residues of the VHH domain are directed against V according to the protocol by Kabat et al ("Sequence of proteins of immunological interest", US Public Health Services, NIH Bethesda, Md., Pub. No. 91)_HThe general numbers given by the fields are numbered. According to this numbering the number of the first row,

-FR1 contains the amino acid residues in positions 1 to 30,

-CDR1 comprises amino acid residues in positions 31 to 35,

-FR2 contains the amino acids in positions 36 to 49,

-CDR2 comprises amino acid residues in positions 50 to 65,

-FR3 contains the amino acid residues at positions 66 to 94,

-CDR3 comprises an amino acid residue in positions 95 to 102, and

-FR4 comprises the amino acid residues at positions 103 to 113.

However, it should be noted that V is well known in the art_HThe total number of amino acid residues in each CDR may vary for the domain and VHH domain, and thus may not correspond to the total number of amino acid residues indicated by Kabat numbering (i.e., one or more positions according to Kabat numbering may not be occupied by the actual sequence, or the actual sequence may contain more amino acid residues than the Kabat numbering allows). This means that although the numbering of the amino acid residues of the VHH domain is based on the numbering according to Kabat, the actual numbering of the amino acid residues in the actual sequence may differ. Since such variations are well known in the art, the individual numbering and assignment of framework regions and CDRs within this sequence can be determined by one of skill in the art without further trouble.

For numbering V_HSubstitution methods for amino acid residues of a domain (these methods can also be applied in a similar manner to VHH domains) are known in the art. However, in this specification, unless the application indicates otherwise, the numbering according to Kabat and applied to VHH domains as described above will be followed by the claims and drawings relating to ISVD as described herein.

The total number of amino acid residues in the VHH domain will typically range from 110 to 120, typically between 112 and 115. However, it should be noted that smaller and longer sequences may also be suitable for the purposes described herein.

Methods for obtaining VHH domains that bind to specific antigens or epitopes have been described earlier, for example in WO2006/040153 and WO 2006/122786. A VHH domain derived from a camelid may be "humanised" by substituting one or more amino acid residues in the amino acid sequence of the original VHH sequence with one or more of the amino acid residues found at corresponding positions in the VH domain of a conventional 4 chain antibody from human. The humanized VHH domain may comprise one or more fully human framework sequences, and in even more particular embodiments, may comprise human framework sequences derived from DP-29, DP-47, DP-51, or portions thereof, optionally in combination with a JH sequence (e.g., JH 5).

The terms "epitope" and "antigenic determinant" are used interchangeably to refer to the portion of a macromolecule (e.g., a polypeptide) that is recognized by an antigen binding molecule (e.g., a conventional antibody or a polypeptide of the invention), and more particularly by the antigen binding site of such molecules. An epitope defines the minimum binding site for an immunoglobulin and thus represents a specific target for the immunoglobulin.

The portion of an antigen binding molecule (e.g., a conventional antibody or a polypeptide disclosed herein) that recognizes an epitope is referred to as a paratope.

The term "biparatopic" (antigen) binding molecule or "biparatopic" polypeptide as used herein shall mean a polypeptide comprising a first and a second ISVD as defined herein, wherein these two variable domains can bind to two different epitopes of an antigen, which epitopes are usually not bound simultaneously by one monospecific immunoglobulin (e.g. a conventional antibody or one ISVD). Biparatopic polypeptides according to the invention comprise variable domains that have different epitope specificities and do not contain complementary pairs of variable domains that bind to the same epitope. Therefore, they do not compete with each other for binding to LRP 5.

A polypeptide (e.g., an immunoglobulin, antibody, ISVD, or antigen binding molecule or fragment thereof in general) that "binds," "specifically binds," or "specifically binds" to a certain epitope, antigen, or protein, has "affinity" for a certain epitope, antigen, or protein (or for at least a portion, fragment, or epitope thereof), and/or is "specific" is said to be "directed to" or "directed against" the epitope, antigen, or protein, or a "binding" molecule for the epitope, antigen, or protein.

In general, the term "specificity" refers to specific antigen bindingThe number of different types of antigens or epitopes that a molecule or antigen binding protein (e.g., immunoglobulin, antibody, ISVD) can bind. The specificity of an antigen binding protein can be determined based on its affinity and/or binding (avidity). By the dissociation equilibrium constant (K) of the antigen to the antigen-binding protein_D) The indicated affinities are measures of the strength of binding between an epitope and an antigen binding site on an antigen binding protein: the K is_DThe smaller the value, the stronger the binding strength between the epitope and the antigen binding molecule (alternatively, the affinity may also be expressed as an affinity constant (K)_A) It is 1/K_D). As will be apparent to the skilled person (e.g. based on further disclosure of the present application), the affinity may be determined in a manner known per se, depending on the specific antigen of interest. A measure of the strength of binding between a binding antigen-binding molecule (e.g., immunoglobulin, antibody, ISVD) and an associated antigen. Binding is the affinity between an epitope and its antigen binding site on an antigen binding molecule and the number of associated binding sites present on the antigen binding molecule.

Typically, an antigen binding molecule (e.g., a polypeptide that specifically binds to LRP5) will have a dissociation constant (K) of 10E-5 to 10E-14 moles/liter (M) or less, and preferably, 10E-7 to 10E-14 moles/liter (M) or less, more preferably, 10E-8 to 10E-14 moles/liter, and even more preferably, 10E-11 to 10E-13_D) (as measured in the Kinexa assay; known in the art), and/or with a binding constant (K11 ME-1) of at least 10E7 ME-1, preferably at least 10E8 ME-1, more preferably at least 10E9 ME-1, e.g., at least 10E11 ME-1_A) And (4) combining. It is generally considered that any K greater than 10E-4M_DValues indicate non-specific binding. Preferably, the antigen binding molecule (e.g., a polypeptide that specifically binds to LRP5) will have a K of less than 500nM, preferably less than 200nM, more preferably less than 10nM, e.g., less than 500pM_DBinding to the desired antigen. Specific binding of an antigen binding protein to an antigen or epitope can be by any suitable means known per se, including for example the assays described herein, Scatchard (Scatchard) assays and/or competitive binding membersAssays such as Radioimmunoassays (RIA), Enzyme Immunoassays (EIA) and sandwich competition assays and different variation assays known per se in the art.

The LRP5 antagonists described herein (which term is used interchangeably with the terms "polypeptide that specifically binds to LRP 5" or "polypeptide that specifically binds to LRP 5") have specificity for LRP5 in that they comprise an ISVD that binds to an epitope of LRP5, but does not or substantially does not cross-react with an epitope having a structure similar to that of LRP5 or having an unrelated structure. Preferably, these LRP5 antagonists have an affinity and/or binding to LRP5, in particular HUMAN LRP5, which is at least 10-fold, preferably at least 100-fold, more preferably at least 1000-fold, even more preferably at least 10000-fold, yet even more preferably at least 100000-fold or at least 1000000-fold stronger than its affinity and/or binding to any other antigen, in particular LRP6, and more in particular HUMAN LRP6 (accession number: UniProtKB-O75581/LRP6_ HUMAN). Most preferably, these LRP5 antagonists are not cross-reactive to LRP6, particularly human LRP 6.

LRP5 antagonists will bind to the human form of LRP5 and preferably also to the counterparts of other species relevant for drug development, i.e. cynomolgus monkey and mouse LRP 5.

As used in this application, the terms "comprises" and variations such as "comprises" and "comprising" may be substituted with the terms "comprising" or "including" or "having". Furthermore, the term "comprising" also expressly encompasses embodiments "consisting of" the enumerated components of this application.

Combination therapy

It is an object of the present invention to provide novel therapies for the treatment or management of various hyperproliferative diseases, in particular various malignancies.

The inventors of the present application have surprisingly found that the use of LRP5 antagonists in combination with anti-PD-1 (programmed cell death protein 1) antibodies has the potential to improve clinical outcomes compared to the use of LRP5 antagonists or anti-PD-1 antibodies alone.

Specifically, in preclinical studies, the inventors tested LRP5 antagonists for immunomodulatory function and anti-tumor activity, either alone or in combination with anti-PD-1 antibodies (see example 1 below). Complete response as determined by histopathological analysis, and massive T cell tumor infiltration were only observable against LRP5 antagonist in combination with anti-PD-1 antibody. FACS analysis of tumor draining lymph nodes further showed that this combination treatment resulted in an increase in the number of activated Dendritic Cells (DCs) in draining lymph nodes. Without wishing to be bound by theory, these findings indicate that combination treatment with LRP5 antagonists and anti-PD-1 antibodies results in inhibition of the Wnt signaling pathway in DCs, which in turn results in upregulation of pro-inflammatory cytokines, cross-priming restoration, and promotion of tumor T cell infiltration and anti-tumor activity.

While various combination therapies are known in the art and are currently under investigation (e.g., in preclinical or clinical trials), there is still a lack of satisfactory therapeutic concepts for the treatment of cancer diseases, in particular, solid tumors such as lung cancer (e.g., NSCLC), melanoma, bladder cancer, and gastrointestinal cancer. Any therapy that shows advantages over standard therapies, such as for example better therapeutic outcome, beneficial effects, excellent efficacy and/or improved tolerability, such as reduced side effects, would therefore represent an important development.

The unexpected results shown in example 1 below indicate that the combination of an LRP5 antagonist (which itself has no therapeutic effect in a tumor model) and an anti-PD-1 antibody (which has only limited therapeutic effect) results in a synergistic (i.e. greater than additive) interaction of these two compounds, providing excellent results so that a complete response can be obtained.

The present invention thus relates to methods for the treatment and/or prevention of hyperproliferative diseases, in particular cancer, comprising the combined administration of an LRP5 antagonist and an anti-PD-1 antibody, each as described herein, and to medical uses, pharmaceutical compositions or combinations and kits comprising these therapeutic agents.

Furthermore, the invention relates to anti-cancer therapies comprising the combined use of an LRP5 antagonist and an anti-PD-1 antibody, each as described herein.

This combination therapy may be given as an unfixed (e.g., free) combination of substances or in a fixed combination (including a backup package).

For the treatment of diseases with oncological properties, a number of anti-cancer agents have been proposed, including target-specific and non-target-specific anti-cancer agents, which can be used as monotherapy or as combination therapy involving more than one agent (e.g. dual or triple combination therapy) and/or which can be combined with radiotherapy (e.g. radiation therapy), radioimmunotherapy and/or surgery. Thus, the combination treatments described herein may be given in addition to other therapeutic agents and/or treatments such as radiotherapy, radioimmunotherapy and surgery.

LRP5 antagonists

Within the meaning of the present invention and all embodiments thereof, a polypeptide that specifically binds to LRP5 (also referred to herein as an LRP5 antagonist) that binds to low density lipoprotein receptor-like 5(LRP5) comprises two or more ISVD that bind to LRP5 at different epitopes, preferably a first domain that blocks the Wnt3a binding site of LRP5 and preferably inhibits Wnt3a driven transcription of a target gene, and/or a second domain that blocks the Wnt1 binding site of LRP5 and preferably inhibits Wnt1 driven transcription of a target gene.

In other words, LRP5 antagonists of the invention include:

-a first ISVD which can specifically bind to LRP5 via an epitope/in a manner leading to inhibition of the Wnt3a signaling pathway, such that Wnt3a driven transcription of a target gene is inhibited, and

a second ISVD that can specifically bind to LRP5 via an epitope/in a manner that results in inhibition of the Wnt1 signaling pathway, such that Wnt 1-driven transcription of the target gene is inhibited.

These molecules are biparatopic binding molecules due to the presence of two ISVD (Wnt1/Wnt3 signaling related) with two domains binding to different epitopes in the LRP5 antagonists described above.

In some embodiments of the invention, the LRP5 antagonist comprises:

- (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

- (b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)。

this particular combination of CDR sequences is for example comprised in LRP5 antagonists referred to hereinafter as LRP5#1, LRP5#2, LRP5#3 and LRP5# 4.

In some embodiments of the invention, the LRP5 antagonist comprises:

- (a) a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3: SRGTSTPSRASGVSRYDY (SEQ ID NO:42), and

- (b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)。

this particular combination of CDR sequences is for example comprised in LRP5 antagonists referred to hereinafter as LRP5#5 and LRP5# 6.

In some embodiments of the invention, the LRP5 antagonist comprises:

- (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

- (b) a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)。

this particular combination of CDR sequences is for example comprised in LRP5 antagonists referred to hereinafter as LRP5#7 and LRP5# 8.

In some embodiments of the invention, the LRP5 antagonist comprises:

- (a) a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3: DRRPRSTGRSGTGSPSTYDY (SEQ ID NO:45), and

- (b) a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3:ETGPYGPPKRDY(SEQ ID NO:48)。

this particular combination of CDR sequences is for example comprised in the LRP5 antagonist referred to hereinafter as LRP5# 9.

As used herein, the terms "first" and "second" in general with respect to these ISVD or domains are only intended to indicate that these domains are two different domains (as they include at least different CDR sequences). Thus, these terms should not be construed to refer to the exact order or sequence of these domains within this polypeptide chain. In other words, isvd (a) and (b) above can be arranged within the polypeptides described herein in the order (a) to (b) or in the order (b) to (a).

In particular, the ISVD (e.g. an ISVD comprising a CDR sequence as defined above) of a polypeptide described herein is a VHH domain, preferably a humanized VHH domain.

In some embodiments of the invention, the LRP5 antagonist comprises a polypeptide having a first ISVD (a) and a second ISVD (b), the first ISVD comprising a VHH domain having a sequence selected from the group consisting of SEQ ID No. 50, SEQ ID No. 51, and SEQ ID No. 62, and the second ISVD comprising a VHH domain having a sequence selected from the group consisting of SEQ ID No. 52, SEQ ID No. 53, and SEQ ID No. 61.

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:50 and the second ISVD comprises the sequence of SEQ ID NO:52 (LRP5# 1).

In some embodiments of the invention, the first ISVD of the LRP5 antagonist comprises the sequence of SEQ ID NO:50 and the second ISVD of the LRP5 antagonist comprises the sequence of SEQ ID NO:61 (LRP5# 2).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:62 and the second ISVD comprises the sequence of SEQ ID NO:52 (LRP5# 3).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:62 and the second ISVD comprises the sequence of SEQ ID NO:61 (LRP5# 4).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:50 and the second ISVD comprises the sequence of SEQ ID NO:53 (LRP5# 5).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:62 and the second ISVD comprises the sequence of SEQ ID NO:53 (LRP5# 6).

In some embodiments of the invention, the first ISVG comprises the sequence of SEQ ID NO:51 and the second ISVD comprises the sequence of SEQ ID NO:52 (LRP5# 7).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:51 and the second ISVD comprises the sequence of SEQ ID NO:61 (LRP5# 8).

In some embodiments, the first ISVD comprises the sequence of SEQ ID NO:51 and the second ISVD comprises the sequence of SEQ ID NO:53 (LRP5# 9).

In preferred embodiments of the invention, the LRP5 antagonist is any one of LRP5#2, LRP5#6 or LRP5#7 as defined by the CDR and/or VHH sequences above.

According to a preferred aspect of the invention, the LRP5 antagonist comprises polypeptides having a first (a) and a second (b) LRP5 that bind to ISVD and a third ISVD (c). Preferably, the LRP5 antagonist comprises first and second ISVD as defined by the CDR sequences above and a third ISVD that is directly or indirectly connected to the first and second ISVD. In some embodiments, the first ISVD is covalently linked to the third ISVD via a peptide linker, and the third ISVD is covalently linked to the second ISVD via a peptide linker. The two linkers may be the same or different linkers. The present application also encompasses the presence of only one linker. As noted above, the terms "first" and "second" do not indicate that they are equal to positions within the polypeptide, and thus from N to C-terminus, the ISVD sequences within the polypeptide may be arranged in the order ISVD (a) - (C) - (b), (a) - [ linker ] - (C) - [ linker ] - (b), (b) - (C) - (a), (b) - [ linker ] - (C) - [ linker ] - (a), (a) - [ linker ] - (C) - (b), (a) - (C) - [ linker ] - (b), (b) - [ linker ] - (C) - (a), (b) - (C) - [ linker ] - (a).

Preferably, the third ISVD (c) is albumin binding ISVD. A non-limiting example of this albumin binding ISVD is the Alb11 domain, which comprises the following CDRs:

CDR(Alb11)1:SFGMS(＝SEQ ID NO:54)

CDR(Alb11)2:SISGSGSDTLYADSVKG(＝SEQ ID NO:55)

CDR(Alb11)3:GGSLSR(＝SEQ ID NO:56)

this results in a group of preferred LRP5 antagonists having the following structure:

FR (a)1-CDR (a)1-FR (a)2-CDR (a)2-FR (a)3-CDR (a)3-FR (a)4- [ optionally linker peptide ] -FR (Alb11)1-CDR (Alb11)1-FR (Alb11)2-CDR (Alb11)2-FR (Alb11)3-CDR (Alb11)3-FR (Alb11)4- [ optionally linker peptide ] -FR (b)1-CDR (b)1-FR (b)2-CDR (b)2-FR (b)3-CDR (b)3-FR (b)4, preferably wherein the CDRs comprise sequences as listed above.

Again, the order of the three isvd (a), (b) and Alb11 is not fixed but shall also comprise polypeptides in which the upper domains are arranged in the following order:

(b)-Alb11-(a)。

in addition, polypeptides having an Alb11 domain (e.g., Alb11- (a) - (b), Alb11- (b) - (a), (a) - (b) -Alb11, or (b) - (a) -Alb11) at the N-or C-terminus of the polypeptide are also intended to be encompassed by the present invention.

In some embodiments of the invention, the LRP5 antagonist includes a polypeptide comprising:

-a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3:SRGTSTPSRASGVSRYDY(SEQ ID NO:42)，

-a second ISVD comprising the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO:48), and

albumin binding ISVD (third ISVD) comprising the following CDR sequences:

CDR1:SFGMS(SEQ ID NO:54)

CDR2:SISGSGSDTLYADSVKG(SEQ ID NO:55)

CDR3:GGSLSR(SEQ ID NO:56)。

this particular combination of CDR sequences is for example comprised in LRP5 antagonists referred to hereinafter as LRP5#1, LRP5#2, LRP5#3 and LRP5# 4.

In some embodiments of the invention, the LRP5 antagonist includes a polypeptide comprising:

-a first ISVD comprising the following CDR sequences:

CDR1:TYVMG(SEQ ID NO:40)

CDR2:AISWSGGSTYYADSVKG(SEQ ID NO:41)

CDR3:SRGTSTPSRASGVSRYDY(SEQ ID NO:42)，

-a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO:48), and

albumin binding ISVD comprising the following CDR sequences:

CDR1:SFGMS(SEQ ID NO:54)

CDR2:SISGSGSDTLYADSVKG(SEQ ID NO:55)

CDR3:GGSLSR(SEQ ID NO:56)。

this particular combination of CDR sequences is included, for example, in the LRP5 antagonists referred to hereinafter as LRP5#5 and LRP5# 6.

In some embodiments of the invention, the LRP5 antagonist includes a polypeptide comprising:

-a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3:DRRPRSTGRSGTGSPSTYDY(SEQ ID NO:45)，

-a second ISVD having the following CDR sequences:

CDR1:IGAMG(SEQ ID NO:46)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO:48), and

albumin binding ISVD comprising the following CDR sequences:

CDR1:SFGMS(SEQ ID NO:54)

CDR2:SISGSGSDTLYADSVKG(SEQ ID NO:55)

CDR3:GGSLSR(SEQ ID NO:56)。

this particular combination of CDR sequences is included, for example, in the LRP5 antagonists referred to hereinafter as LRP5#7 and LRP5# 8.

In some embodiments of the invention, the LRP5 antagonist includes a polypeptide comprising:

-a first ISVD comprising the following CDR sequences:

CDR1:RYAVA(SEQ ID NO:43)

CDR2:AITWSSGRIDYADSVKG(SEQ ID NO:44)

CDR3:DRRPRSTGRSGTGSPSTYDY(SEQ ID NO:45)，

-a second ISVD comprising the following CDR sequences:

CDR1:INAMG(SEQ ID NO:49)

CDR2:AVSSGGSTYYVDSVKG(SEQ ID NO:47)

CDR3: ETGPYGPPKRDY (SEQ ID NO:48), and

albumin binding ISVD comprising the following CDR sequences:

CDR1:SFGMS(SEQ ID NO:54)

CDR2:SISGSGSDTLYADSVKG(SEQ ID NO:55)

CDR3:GGSLSR(SEQ ID NO:56)。

this particular combination of CDR sequences is for example comprised in the LRP5 antagonist referred to hereinafter as LRP5# 9.

In some embodiments, the ISVD's in the LRP5 antagonists above as defined by the CDR sequences of the ISVD's are arranged such that the albumin-binding ISVD connects the first and second ISVD, either directly or indirectly (e.g., via (a) a linker peptide).

In a preferred embodiment, the sequence of Alb11 ISVD mentioned above is as follows:

EVQLVESGGGLVQPGNSLRLSCAASGFTFSSFGMSWVRQAPGKGLEWVSSISGSGSDTLYADSVKGRFTISRDNAKTTLYLQMNSLRPEDTAVYYCTIGGSLSRSSQGTLVTVSS

(═ Alb11 domain;: SEQ ID NO:60)

The CDR sequences mentioned above are summarized in tables 1A, 1B and 1C:

table 1A: CDR sequences of ISVD that interfere with Wnt3a signaling:

table 1B: CDR sequences of ISVD that interfere with Wnt1 signaling:

table 1C: CDR sequences of ISVD that bind to serum albumin (Alb11 domain):

the three preferred LRP5 antagonists described herein are as follows:

first preferred LRP5 antagonist (LRP5# 6): a polypeptide comprising:

-a first (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 62;

-albumin binding ISVD comprising the amino acid sequence as shown in SEQ ID No. 60;

-a second (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 53;

in this order, or in an order that the three domains change above.

Second preferred LRP5 antagonists (LRP5# 7): a polypeptide comprising:

-a first (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 51;

-albumin binding ISVD comprising the amino acid sequence as shown in SEQ ID No. 60;

-a second (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 52;

in this order, or in an order that the three domains change above.

Third preferred LRP5 antagonists (LRP5# 2): a polypeptide comprising:

-a first (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 50;

-albumin binding ISVD comprising the amino acid sequence as shown in SEQ ID No. 60;

-a second (LRP5 binding) ISVD comprising the amino acid sequence as shown in SEQ ID No. 61;

in this order, or in an order that the three domains change above.

In an even more specific preferred embodiment, the albumin-binding ISVD is located between two ISVD-binding LRP 5.

The sequences of the VHHs mentioned above are summarized in tables 2A, 2B and 2C:

table 2A: VHH sequence of ISVD that interferes with Wnt3a signaling:

table 2B: VHH sequence of ISVD that interferes with Wnt1 signaling:

table 2C: sequence of ISVD binding to serum albumin (Alb11 domain):

in preferred embodiments of the invention, the LRP5 antagonists comprise a sequence selected from SEQ ID NOs 57, 58, and 59 (these preferred LRP5 antagonists are also referred to herein as LRP5#6, LRP5#7, and LRP5#2, respectively), wherein the precise amino acid sequence can be taken from table 2D below:

table 2D:

the manufacture and therapeutic use of the above LRP5 antagonists is disclosed in WO2018/220080a 1. In particular, this document provides a full disclosure of methods of making LRP5 antagonists for use in the present invention.

anti-PD-1 antibodies

anti-PD-1 antibodies (also referred to herein as "PD-1 antibodies") and all embodiments thereof within the meaning of the present invention are compounds that inhibit the interaction of PD-1 with its ligand. Preferably, the anti-PD-1 antibody is a humanized or fully human anti-PD-1 antibody. Any of these antibodies may be recombinant human antibodies.

The PD-1 gene encodes a 55kDa type I transmembrane protein, which is part of the Ig gene superfamily (Agata et al (1996) Int Immunol.8: 765-72). The complete PD-1 sequence can be found under genbank accession number U64863. Although structurally similar to CTLA-4, PD-1 lacks the MYPPY matrix (SEQ ID NO:39), which is important for B7-1 and B7-2 binding.

PD-1 is an inhibitory member of the expanded CD28/CTLA-4 family of T cell regulatory factors. Other members of the CD28 family include CD28, CTLA-4, ICOS and BTLA. PD-1 is proposed to exist as a monomer, lacking the unpaired cysteine residue characteristic of other CD28 family members. PD-1 is expressed on activated B cells, T cells and monocytes (Okazaki et al (2002) CurrOpinImmunol 14: 391779-82; Bennett et al (2003) J. Immunol.170: 711-8). Two ligands for PD-1, PD-L1(B7-H1) and PD-L2(B7-DC), have been identified which have been shown to bind to PD-1, i.e., to down-regulate T cell activation (Freeman et al (2000) J.Exp.Med.192: 1027-34; Carter et al (2002) Eur.J.Immunol.32: 634-43). Both PD-L1 and PD-L2 are B7 homologs that bind to PD-1. PD-L1 is abundantly present in a variety of human cancers (Dong et al (2002) nat. Med.8: 787-9).

PD-1 is known as an immunosuppressive protein that negatively regulates TCR signaling (Ishida, Y. et al (1992) EMBO J.11: 3887-3895; Blank, C. et al (2006) Immunol. immunother.56(6): 739-745). The interaction between PD-1 and PD-L1 can serve as an immune checkpoint that can lead to, for example, a reduction in tumor-infiltrated lymphocytes, a reduction in proliferation mediated by T cell receptors, and/or immune evasion of Cancer cells (Dong et al (2003) J. mol. Med.81: 281-7; Blank et al (2005) Cancer Immunol. Immunother.54: 307-314; Konishi et al (2004) Clin. Cancer Res.10: 5094-100). Immunosuppression can be reversed by inhibiting local interaction of PD-1 with PD-L1 or PD-L2; the effect is additive when blocking the interaction of PD-1 with both PD-L1 and PD-L2 (Iwai et al (2002) Proc. nat' l.Acad.Sci USA 99: 12293-7; Brown et al (2003) J.Immunol.170: 1257-66).

In one aspect of the invention, an anti-PD-1 antibody is defined by any one of antibodies PD1-1, PD1-2, PD-1-3, PD1-4 and PD1-5 whose sequences are as shown in table 3 by means of SEQ ID numbers, wherein VH represents a heavy chain variable domain, VL represents a light chain variable domain, HC represents a (full length) heavy chain and LC represents a (full length) light chain:

table 3: SEQ ID NOs of CDR, VH, VL, HC and LC sequences

And wherein the amino acid sequences (and sequence names) of the SEQ ID numbers are shown in table 4 below:

table 4:

specifically, the anti-PD-1 antibody molecules described herein comprise:

(a) heavy chain CDRs comprising the amino acid sequences of SEQ ID NO 1(HCDR1), SEQ ID NO 2(HCDR2) and SEQ ID NO 3(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO 4(LCDR1), SEQ ID NO 5(LCDR2) and SEQ ID NO 6(LCDR 3); or

(b) Heavy chain CDRs comprising the amino acid sequences of SEQ ID NO 7(HCDR1), SEQ ID NO 8(HCDR2) and SEQ ID NO 9(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO 10(LCDR1), SEQ ID NO 11(LCDR2) and SEQ ID NO 12(LCDR 3); or

(c) Heavy chain CDRs comprising the amino acid sequences of SEQ ID NO 13(HCDR1), SEQ ID NO 14(HCDR2) and SEQ ID NO 15(HCDR3) and light chain CDRs comprising the amino acid sequences of SEQ ID NO 16(LCDR1), SEQ ID NO 17(LCDR2) and SEQ ID NO 18(LCDR 3).

In some embodiments, the anti-PD-1 antibody molecule includes a heavy chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 19, 21, 23, 25, and 27.

In some embodiments, the anti-PD-1 antibody molecule includes a light chain variable domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOs 20, 22, 24, 26, and 28.

In some embodiments, the anti-PD-1 antibody molecule comprises:

(a) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 19 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 20,

(b) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 21 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 22,

(c) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 23 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 24,

(d) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO. 25 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO. 26, or

(e) A heavy chain variable domain comprising the amino acid sequence of SEQ ID NO 27 and a light chain variable domain comprising the amino acid sequence of SEQ ID NO 28.

In some embodiments, the anti-PD-1 antibody comprises:

(a) a heavy chain comprising the amino acid sequence of SEQ ID NO. 29 and a light chain comprising the amino acid sequence of SEQ ID NO. 30,

(b) a heavy chain comprising the amino acid sequence of SEQ ID NO. 31 and a light chain comprising the amino acid sequence of SEQ ID NO. 32,

(c) a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34,

(d) heavy chain comprising the amino acid sequence of SEQ ID NO 35 and light chain comprising the amino acid sequence of SEQ ID NO 36, or

(e) A heavy chain comprising the amino acid sequence of SEQ ID NO 37 and a light chain comprising the amino acid sequence of SEQ ID NO 38.

In a preferred embodiment, the anti-PD-1 antibody PD 1-1.

In a preferred embodiment, the anti-PD-1 antibody PD 1-2.

In a preferred embodiment, the anti-PD-1 antibody PD 1-3.

In a preferred embodiment, the anti-PD-1 antibody PD 1-4.

In a preferred embodiment, the anti-PD-1 antibody PD 1-5.

In one aspect, the invention provides a method of treating and/or preventing a hyperproliferative disease, preferably a cancer, comprising administering to a patient in need thereof a therapeutically effective amount of an LRP5 antagonist (e.g., any of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VH/VL sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) and a therapeutically effective amount of an anti-PD-1 antibody (e.g., any of PD1-1, PD1-2, PD1-3, PD1-4, PD 1-5) and a therapeutically effective amount of an anti-PD-1 antibody. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In a further aspect, the invention provides a combination of an LRP5 antagonist described herein (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) with an anti-PD-1 antibody described herein (e.g. any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4), particularly for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably a cancer, wherein the method comprises administering to a patient in need thereof a therapeutically effective amount. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the invention relates to an LRP5 antagonist described herein (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the method comprises administering to a patient in need thereof an effective amount of the antagonist 5 in combination with an anti-PD-1 antibody described herein (e.g. any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4). In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the invention relates to an anti-PD-1 antibody described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4) for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, wherein the method comprises administering to a patient in need thereof a therapeutically effective amount of the anti-PD-1 antibody in combination with any one of the LRP5 antagonists described herein (e.g., LRP5#1, LRP5#2, LRP5#3, 737 5#4, LRP5#5, LRP 25 #6, 733 #7, LRP5#8, LRP5 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c). In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the invention relates to a kit comprising, in one or more containers:

a first pharmaceutical composition or dosage form comprising an LRP5 antagonist described herein (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c), and optionally one or more pharmaceutically acceptable carriers, excipients and/or vehicles, and

a second pharmaceutical composition or dosage form comprising an anti-PD-1 antibody as described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4), and optionally one or more pharmaceutically acceptable carriers, excipients, and/or vehicles;

and optionally a package insert comprising printed instructions.

In a preferred embodiment of the kits of the invention, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In a preferred embodiment of the kits of the invention, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In a preferred embodiment of the kits of the invention, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

Preferably, the package insert comprises printed instructions for the simultaneous, concurrent, sequential, alternating or separate treatment and/or prevention of a hyperproliferative disease, in particular cancer, in a patient in need thereof as described herein.

In another aspect, the present invention relates to the above-mentioned kit for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, as described herein.

In another aspect, the present invention relates to a pharmaceutical composition comprising:

an LRP5 antagonist described herein (e.g., any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c),

an anti-PD-1 antibody described herein (e.g., any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4), and

optionally, one or more pharmaceutically acceptable carriers, excipients and/or vehicles.

In a preferred embodiment of the pharmaceutical composition of the invention, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments of the pharmaceutical composition, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments of the pharmaceutical composition, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the invention relates to the use of an LRP5 antagonist described herein (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) for the preparation of a pharmaceutical composition for use in a method of treatment and/or prevention of an hyperproliferative disease, preferably cancer, as described herein, wherein the LRP 6342 antagonist is to be used in combination with a PD-1 antibody as described herein (e.g. any one of PD1-1, PD1-2, PD 6345-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4). In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the invention relates to the use of a PD-1 antibody as described herein (e.g. any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4) in the manufacture of a pharmaceutical composition for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably cancer, as described herein, wherein the PD-1 antagonist is to be used in combination with an LRP5 antagonist described herein (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5) as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c). In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In a further aspect, the invention relates to the use of an LRP5 antagonist (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) and a PD-1 antibody (e.g. any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4) for the preparation of a pharmaceutical composition for use in a method for the treatment and/or prevention of a hyperproliferative disease, preferably a cancer, as described herein. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

In another aspect, the present invention relates to a combination, a pharmaceutical composition or a kit according to the invention, each as described herein, comprising, consisting of or consisting essentially of: LRP5 antagonists (e.g. any one of LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 as defined by the CDR and/or VHH sequences of tables 1a, 1b, 1c, 2a, 2b, 2 c) and anti-PD-1 antibodies (e.g. any one of PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 as defined by the CDR and/or VH/VL sequences of tables 3 and 4) for use in a method of treatment and/or prevention of a hyperproliferative disease, preferably a cancer, as described herein. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 29 and a light chain comprising the amino acid sequence of SEQ ID NO 30. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 31 and a light chain comprising the amino acid sequence of SEQ ID NO 32. In preferred embodiments, the LRP5 antagonist comprises the amino acid sequence of SEQ ID NO 57, SEQ ID NO 58, or SEQ ID NO 59 and the PD-1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO 33 and a light chain comprising the amino acid sequence of SEQ ID NO 34.

The permutations of the examples for the LRP5 antagonists LRP5#1, LRP5#2, LRP5#3, LRP5#4, LRP5#5, LRP5#6, LRP5#7, LRP5#8, LRP5#9 and the examples for the PD-1 antagonists PD1-1, PD1-2, PD1-3, PD1-4, PD1-5 result in all specific combinations that should be considered as explicitly disclosed and all of the examples of the invention and the combinations, compositions, kits, methods, uses and compounds thereof for use, including methods/dosing regimens applying specific administrations as specified below and/or methods for treating specific cancers as specified below.

Routes of administration for LRP5 antagonists and/or anti-PD 1 antibodies as described herein include, but are not limited to, parenteral (e.g., intramuscular, intraperitoneal, intravenous, transdermal or subcutaneous injection or implant), oral, parenteral, nasal, vaginal, rectal, or topical administration. In a preferred embodiment, the route of administration is intravenous administration, in particular intravenous infusion or injection. The compounds of the present invention may be formulated, alone or together, into suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, excipients, and/or vehicles suitable for each route of administration. More preferably, the formulations include solid, semi-solid, or liquid dosage forms, such as lyophilizates, liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, liposomes, and suppositories. The preferred mode depends on the intended mode of administration and therapeutic application. Particularly preferred embodiments include liquid formulations and lyophilizates. In the case of a lyophilizate, the lyophilizate can be reconstituted in a liquid, preferably water.

As described herein, administration of an anti-PD-1 antibody can be at a dose of about 0.1 to 30mg/kg of patient body weight, e.g., about 0.5 to 25mg/kg of patient body weight, about 1 to 20mg/kg of patient body weight, about 2 to 5mg/kg of patient body weight, or about 3mg/kg of patient body weight, e.g., by injection (e.g., subcutaneously or intravenously).

In some embodiments, the anti-PD-1 antibody is administered at a dose of about 10 to 20mg/kg of patient body weight every two weeks. The antibody molecule can be administered by intravenous infusion at a rate greater than 20mg/min, e.g., 20 to 40mg/min, and typically greater than or equal to 40mg/min to achieve about 35 to 440mg/m²Usually about 70 to 310mg/m²And more typically, about 110 to 130mg/m²The dosage of (a). In some embodiments, about 110 to 130mg/m²Up to a concentration of about 3mg/kg of patient body weight. In other embodiments, the antibody molecule can be administered by intravenous infusion at a rate of less than 10mg/min, e.g., less than or equal to 5mg/min, to achieve about 1 to 100mg/m²E.g., about 5 to 50mg/m²About 7 to 25mg/m²Or about 10mg/m²The dosage of (a). In some embodiments, the antibody is infused for a period of about 30 min.

A preferred dosage regimen for an anti-PD-1 antibody described herein comprises administering 1mg/kg of the patient's body weight or 3mg/kg of the patient's body weight via intravenous administration, and the antibody is given every three weeks or every four weeks.

The LRP5 antagonists described herein or compositions comprising the same may be administered, for example, intravenously (i.v.), subcutaneously (s.c.), intramuscularly (i.m.), intraperitoneally (i.p.), transdermally, orally, sublingually (e.g., in the form of a sublingual tablet, spray or drop placed sublingually and absorbed through the mucosa into the sublingual capillary network), (intranasally) nasally (e.g., in the form of a nasal spray and/or as an aerosol), topically, by means of a suppository, by inhalation, or in any other suitable manner in effective amounts or dosages.

The LRP5 antagonists described herein will generally be administered in amounts and doses between 0.005 and 20.0mg per kilogram of patient body weight, preferably between 0.05 and 10.0 mg/kg/dose, and more preferably between 0.5 and 10 mg/kg/dose, but may vary depending on, among other things, the particular disease, disorder or condition to be treated, the potency of the particular LRP5 antagonist to be used, the particular route of administration used, and the particular pharmaceutical formulation or composition. Thus, in some cases it may be sufficient to use less than the minimum dose given above, whereas in other cases it may be necessary to exceed the upper limit. When administered in large amounts, it is desirable to divide them equally into a number of smaller doses distributed throughout the day.

It should be noted that the dosage amount may vary with the type and severity of the condition to be alleviated. It will be further appreciated that for any particular individual, the specific dosage regimen will be adjusted over time according to the individual requirements and the professional judgment of the person administering the composition or the monitoring composition.

The LRP5 antagonist and the anti-PD 1 antibody as described herein can be administered in therapeutically effective amounts in a single dose or in divided doses administered at appropriate time intervals. A therapeutically effective amount refers to an amount that is effective at the dosages and time periods necessary to achieve the desired therapeutic result and is the minimum amount necessary to prevent, alleviate or treat the disease or disorder. The therapeutically effective amount of a compound described herein can vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects. A therapeutically effective dose preferably inhibits a measurable parameter, such as inhibiting the rate of tumor growth by at least about 20%, more preferably by at least about 40%, even more preferably by at least about 60%, and still more preferably by at least about 80%, relative to an untreated individual or relative to a previous untreated cycle of the same individual to be treated.

The active compounds may be administered at these doses, which are therapeutically effective in monotherapy, or at doses lower or higher than those used in monotherapy, but which when combined result in the desired (co-) therapeutically effective amount. This may be useful, for example, to avoid, limit or reduce any undesirable side effects associated with the use of one or more substances or the principles of achieving a desired medicinal or therapeutic effect when these substances are used in their usual amounts, while still achieving the desired therapeutic effect.

The amount of a compound described herein required for use in treatment can be adapted to the particular compound selected, the route of administration, the nature of the condition being treated and the age and condition of the patient and will ultimately be at the discretion of the attendant physician or clinician. Likewise, the dosage of the compounds described herein can be adjusted depending on the target cell, tumor, tissue, implant, or organ.

The required dose of the LRP5 antagonist or the anti-PD-1 antibody, both as described herein, can be administered as a fixed amount per administration or as a bolus to achieve a set blood concentration in the patient.

Within the present invention, the LRP5 antagonist and the anti-PD 1 antibody are known to be administered either dependently (i.e., mixed together in one composition) or separately (i.e., as separate compositions) in a formulation wherein such administration provides therapeutically effective concentrations of the two compounds in the body of the patient. The latter also applies to cocktail therapies, such as the administration of three or more active agents. In other words, the LRP5 antagonist and the anti-PD 1 antibody can be administered as part of the same pharmaceutical composition/dosage form, or preferably, in different pharmaceutical compositions/dosage forms. With respect to administration in different pharmaceutical compositions/dosage forms, it is understood that simultaneous, concurrent, sequential or alternating administration of the active agents or components is contemplated for such administration in accordance with the present invention.

The term "simultaneously" (also referred to herein as "concomitantly") refers to the administration of both compounds/compositions substantially simultaneously.

Parallel administration includes administration of the active agents within the same general time period, e.g., the same day but not necessarily simultaneously.

Sequential administration includes administration of one agent using one or more doses over a first time period (e.g., over hours, days, or weeks), followed by administration of another agent using one or more doses over a second time period (e.g., over hours, days, or weeks). Overlapping schedules, which include different days within the treatment cycle, may also be employed, not necessarily according to a regular sequential administration of the active agents. Alternatively, continuous administration is also contemplated, with the second administration step occurring immediately upon completion of administration of the first compound. The skilled person knows how to determine the completion of the first administration step, whereby it is possible to determine a point in time suitable for starting the second administration step.

The alternating administration includes administering one agent over a period of time, for example over a period of hours, days, or weeks, followed by administering another agent over a subsequent period of time, for example over a period of hours, days, or weeks, and then repeating the pattern for one or more cycles, wherein the total number of repetitions depends on the selected dosage regimen.

Variations of these general guidelines may also be employed, for example, depending on the agent used and the condition of the individual.

In a preferred embodiment of the invention, in a method according to the invention, each of the LRP5 antagonist and the anti-PD 1 antibody as described herein is administered simultaneously or concurrently (e.g., by intravenous infusion or subcutaneously) over a first period, followed by administration of the anti-PD 1 antibody (e.g., by intravenous infusion or subcutaneously) and no LRP5 antagonist over a second period.

In some embodiments, the first cycle is 3 or 6 weeks when the LRP5 antagonist and PD1 antibody are administered every three weeks.

In some embodiments, the first cycle is 4 or 8 weeks when the LRP5 antagonist and PD1 antibody are administered every four weeks.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-1, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-1, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#2 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#2 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

In another preferred embodiment of the invention, the LRP5 antagonist and the anti-PD 1 antibody as described herein are administered together (by intravenous infusion or subcutaneously, simultaneously or concurrently) and then the anti-PD 1 antibody every three weeks (e.g., 3 or 6 weeks) during a first cycle, e.g., every three weeks (e.g., by intravenous infusion or subcutaneously) during a second cycle. For example, the LRP5 antagonist and anti-PD 1 antibody are administered (i) within week 1 or (ii) concurrently or concurrently (e.g., by intravenous infusion or subcutaneous) within weeks 1 and 4, and then PD1 antibody is administered, e.g., within weeks 7, 10, and any subsequent third week (weeks 13, 16, etc.) until the end of treatment. In the case of option (i), the PD1 antibody has been administered alone within week 4 (i.e., instead of in combination with the LRP5 antagonist as in option (ii)).

Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#2 and anti-PD-1 antibody being PD1-1, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-1, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-1, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#2 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-2, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#2 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#6 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy. Particularly preferably, this administration schedule is employed with LRP5 antagonist being LRP5#7 and anti-PD-1 antibody being PD1-3, even more preferably for the treatment of gastrointestinal, melanoma, bladder or lung cancer (including gastrointestinal, melanoma, bladder and lung cancer that are refractory or resistant to checkpoint inhibitor therapy) or any solid tumor that is refractory or resistant to checkpoint inhibitor therapy.

In another preferred embodiment of the invention, the LRP5 antagonist and the anti-PD 1 antibody as described herein are administered together (by intravenous infusion or subcutaneously, simultaneously or concurrently) and then the anti-PD 1 antibody every four weeks (e.g., by intravenous infusion or subcutaneously) during a first cycle, e.g., every four weeks (e.g., by intravenous infusion or subcutaneously) during a second cycle. For example, the LRP5 antagonist and anti-PD 1 antibody are administered (i) within week 1 or (ii) concurrently or concurrently (e.g., by intravenous infusion or subcutaneous) within weeks 1 and 5, and then the PD1 antibody is administered, e.g., within weeks 9, 13 and any subsequent weeks (weeks 17, 21, etc.) until the end of treatment. In the case of option (i), the PD1 antibody has been administered alone within week 5 (i.e., instead of in combination with the LRP5 antagonist as in option (ii)).

Preferably, an LRP5 antagonist as described herein (e.g., at a dose of about 0.5 to 10mg/kg of patient body weight) and an anti-PD 1 antibody as described herein (e.g., at a dose of any one of 2, 3,4, or 5mg/kg of patient body weight) are administered together (simultaneously or concurrently by intravenous infusion or subcutaneously) and then an anti-PD 1 antibody every three or four weeks (e.g., corresponding to 1 or 2 doses) over a first period, e.g., every three or four weeks (e.g., by intravenous infusion or subcutaneously) over a second period.

In some embodiments of the invention, an LRP5 antagonist and an anti-PD 1 antibody as described herein are administered together (either simultaneously or concurrently by intravenous infusion or subcutaneously) every three or four weeks (e.g., corresponding to 1 or 2 doses) over a first period and then an anti-PD 1 antibody is administered (e.g., by intravenous infusion or subcutaneously) weekly, every other week, every three weeks, or monthly over a second period.

Depending on the disease to be treated, the combination therapy as defined herein may be used alone or in further combination with one or more additional therapeutic agents, in particular chemotherapeutic agents or therapeutically active compounds selected from inhibiting angiogenesis, signal transduction pathways or mitotic checkpoints in cancer cells.

The additional therapeutic agent may be administered concurrently with the LRP5 antagonist and/or the PD1 antibody, optionally as a component of the same pharmaceutical preparation, or before or after administration of the LRP5 antagonist and/or the PD1 antibody.

This/these additional therapeutic agent(s) may be selected from (and are not limited to) the following:

immunotherapeutic agents, e.g. modulators of the following checkpoint inhibitors: TIM3, PD-L1, PD-L2, CTLA-4, VISTA, BTLA, TIGIT, CD160, LAIR1, 2B4, CEACAM;

a cancer vaccine;

a DNA damaging agent;

an inhibitor of angiogenesis;

inhibitors of signal transduction pathways;

inhibitors of mitotic checkpoints; and

hormones, hormone analogues and anti-hormones (e.g. tamoxifen (tamoxifen), toremifene (toremifene), raloxifene (raloxifene), fulvestrant (fulvestrant), megestrol acetate (megestrol acetate), flutamide (flutamide), nilutamide (nilutamide), bicalutamide (bicalutamide), amicarbazone (aminoglutethimide), cyproterone acetate (cyproterone acetate), finasteride (finasteride), butolozoline acetate (buselin acetate), flucortisone (fluucortisone), fluorometholone (fluoromesterone), medroxyprogesterone (medroxyprogesterone), octreotide (octreotide)), aromatase inhibitors (e.g. anastrozole), letrozole (letrozole), Rilotrimazole (RH), growth factor (leuproletasone), growth factor (PDGF), growth factor (e.g. leuproletasone), growth factor (e) and growth factor (e.g. leuproletasone), growth factor (e.g. growth factor (leuproletasone), growth factor (e.g. leuproletasone), growth factor (e), growth factor (e.g. leuproletasone), leuproletasone (e), and/or a, "Fibroblast Growth Factor (FGF)", "Vascular Endothelial Growth Factor (VEGF)", "Epidermal Growth Factor (EGF)", "insulin-like growth factor (IGF)", "human epidermal growth factor (HER, e.g. HER2, HER3, HER 4)" and "Hepatocyte Growth Factor (HGF)", inhibitors such as "growth factor" antibodies, "growth factor receptor" antibodies and tyrosine kinase inhibitors, e.g. cetuximab (cetuximab), gefitinib (gefitinib), imatinib (imatinib), lapatinib (lapatinib), bosutinib (bosutinib) and trastuzumab (trastuzumab)); antimetabolites (e.g., antifolates such as methotrexate, raltitrexed, pyrimidine analogs such as 5-fluorouracil (5-FU), capecitabine and gemcitabine, purine and adenosine analogs such as mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, ara-C, fludarabine); antitumor antibiotics (e.g. anthracenes)Cyclic drugs such as doxorubicin (doxorubicin), doxorubicin (doxil) (pegylated liposomal doxorubicin hydrochloride, daunorubicin (myocet) (non-pegylated liposomal doxorubicin), daunorubicin (daunorubicin), epirubicin (epirubicin), and idarubicin (idarubicin), mitomycin-C (mitomycin) -C), bleomycin (bleomycin), actinomycin (dactinomycin), plicamycin (plicamycin), streptozotocin (streptozocin)); platinum derivatives (e.g., cisplatin (cissplatin), oxaliplatin (oxaliplatin), carboplatin (carboplatin)); alkylating agents (e.g., estramustine (estramustine), mechlorethamine (meclorethamine), melphalan (melphalan), chlorambucil (chlorambucil), busulfan (busulfan), dacarbazine (dacarbazin), cyclophosphamide (cyclophophamide), ifosfamide (ifosfamide), temozolomide (temozolomide), nitrosoureas such as carmustine (carmustin) and lomustine (lomustin), thiotepa (thiotepa)); antimitotic agents (e.g., Vinca alkaloids (Vinca alkaloids) such as vinblastine (vinblastine), vindesine (vindesin), vinorelbine (vinorelbin), and vincristine (vinchristine), and taxanes such as paclitaxel (paclitaxel), docetaxel (docetaxel)); angiogenesis inhibitors (e.g., tasquinimod), tubulin inhibitors; DNA synthesis inhibitors (e.g., sapacitabine (sapacitabine)), PARP inhibitors, topoisomerase inhibitors (e.g., epipodophyllotoxin (epipodophyllotoxin) such as etoposide (etoposide) and etopohos (etophos), teniposide (teniposide), oxamine (amsacrin), topotecan (topotecan), irinotecan (irinotecan), mitoxantrone (mitoxantrone)), serine/threonine kinase inhibitors (e.g., PDK 1 inhibitors, Raf inhibitors, A-Raf inhibitors, B-Raf inhibitors, C-Raf inhibitors, mTOR inhibitors, mTOR 1/2 inhibitors, PI3K inhibitors, PI3K alpha inhibitors, dual mTOR/PI3K inhibitors, STK 33 inhibitors, AKT inhibitors, PLK 1 inhibitors, CDK inhibitors, tyrosine kinase inhibitors (e.g., PTK 2/protein K mutual inhibitors), protein activators (e.g., protein IAP inhibitors), Mcl-1, MDM2/MDMX), MEK inhibitors (e.g. pimatinib), ERK inhibitors, FLT3 inhibitionAgents (e.g., quinazatinib), BRD4 inhibitors, IGF-1R inhibitors, TRAILR2 agonists, Bcl-xL inhibitors, Bcl-2 inhibitors (e.g., venetoclax), Bcl-2/Bcl-xL inhibitors, ErbB receptor inhibitors, BCR-ABL inhibitors, Src inhibitors, rapamycin (rapamycin) analogs (e.g., everolimus), sirolimus (temsirolimus), diphospholimus (ridaforolimus), sirolimus (sirolimus)), androgen synthesis inhibitors (e.g., abiraterone (abatarone), TAK-700), androgen receptor inhibitors (e.g., enzalutamide (enzalutamide), ARN-509), immunotherapy (e.g., sipuleucel-T (sipuluceucine-T)), telocidine inhibitors (e.g., SGI 110, temozoloside (HDAC)), and lipoxin inhibitors (e.g., vorinoside (s (doxorubin)), and the like, Entinostat (entinostat), prunosporal (pracinostat), panobinostat (panobinostat)), ANG1/2 inhibitors (e.g., terbinafine (trebanib)), CYP17 inhibitors (e.g., galidone (galeterone)), radiopharmaceuticals (e.g., radium-223, radium chloride-223 (alpharadin)), immunotherapeuticals (e.g., poxvirus-based vaccines, ipilimumab (ipilimumab), immune checkpoint inhibitors), and various chemotherapeutic agents such as amifostine (amifostin), anagrelide (anagrelid), clodronate (clodronat), filgrastim (grastimulin), interferon alpha, leucovorin (leucovorin), rituximab (rituximab), procarbazine (carpronine), levamisole (levamisole), sodium mestamide (mestam), mestam (mestam), mitotane (mitotane), mitomycin (mitomycin), and milrinone (phenomycin); 2-chlorodeoxyadenosine, 2-fluorodeoxycytidine, 2-methoxyestradiol, 2C4, 3-Allergenin (3-alethine), 131-I-TM-601, 3CPA, 7-ethyl-10-hydroxycamptothecin, 16-aza-epothilone B, ABT-199, ABT-263/Navitoxix (navitoclax), ABT-737, A105972, A204197, aldesleukin (aldesleukin), alicet (alisertib)/MLN8237, aliretinic acid (alitretinoin), isotretinoin-7 (allovitexin-7), altretamine (altretamine), alvocidib (alvocidib), amonafide (amonafide), anthrapyrazole (anthrazole), AG-7, AP-5280, apalone (apazimine), arabinogalactamine (apazimine), arabinogalactine (arabinogalactamine), arabinogalactine (arabinogalactine), arabinoside (arabinogalactine), arabinogalactine (arabinoside), arabinogalactine (arabinogalactine), arabinoside (arabinogalactine), arabinoside (arabinoside), arabinogalactine (arabinogalactine), arabinoside), or a-L-R-7, L-7, L-7, L-L, Arzoxifene (ar)zoxifene), atamestane, atrasentan (atrasentan), auristatin PE (auristatin PE), AVLB, AZ10992, ABX-EGF, AMG-479 (ganitatumab), AMG-232, AMG-511, AMG 2520765, AMG 2112819, ARRY 162, ARRY 438162, ARRY-300, ARRY-142886/AZD-6244 (Semetinib), ARRY-704/AZD-8330, ATSP-7041, AR-12, AR-42, AS-703988, AXL-1717, AZD-1480, AZD-4547, AZD-8055, AZD-5363, AZD-6244, AZD-7762, ARQ 680, AS-706 (primeveratinib)), azastatin (azaphenacetin), azalide (azalide), azalide (saratin), azalide (20145, azalide), azalidine (azalidine B-20145), azalidine (azalidine B-8330), and azalidine, Bararetinib (barasertib)/AZD1152, BAY-43-9006, BAY 80-6946, BBR-3464, BBR-3576, bevacizumab (bevacizumab), BEZ-235/Daxotinib (dacylisib), bicitricrondate (biricotide dicrate), Birinapant (birinapag), BCX-1777, BKM-120/buparib (buparlisib), bleomycin (bleocin), BLP-25, BMS-184476, BMS-247550, BMS-188797, BMS-275291, BMS-663513, BMS-754807, BI-1350, BI-7787, BIBW 2992/afatinib (BInininib), BF 1120/nintedanib (bledarabib), BI-256727, BI 6727/6727, BII 6757, BII-6757, BIIB, 853520, 3636363682, bleomycin, 363682, bleomycin, bleb, Brivarb (brivarinib), bryozoacin-1, bortezomib (bortezomib), bromomycin (brosplallin), busulfan, BYL-719/aplisib (alpelisib), CA-4 prodrug, CA-4, cabazitaxel (cabazitaxel), cabozantinib (cabozantinib), capped cells (Capcell), calcitriol (calcitriol), canertib (canertinib), glycyrrhizamide (canfosfamide), capecitabine, carboplatin (carbopyphthalatlopatin), CCI-779, CC-115, CC-223, CEP-701, CEP-751, CBT-1cefixime (CBT-1cefixime), homoharringtonine (cefixoripine (ceftriaxone), celecoxib (celecoxib), celecoxib (49655), interleukin (49877), gibberellin (CGMCC 877), CGMCC-3, BCT-1, CET-1, CENTE (CGMCC-1, CENTC-3), CENTC-3, CET-3, CENTC-3, BCE (CGMCC-3), BCE, BCG-3, BCG-D (CGMCC-3), CGMCC-3, BCE, BCG-3, BCE (BCE, BCG-D-3, BCE, BCC-D (CGM-D (CGI, C-D-3, C-D-, CDA-II, CDC-394, CKD-602, CKI-27, clofarabine (clofarabine), colchicine (colchicin)Compstatin A4(combretastatin A4), COT inhibitor, CHS-828, CH-5132799, CLL-Thera, CMT-3 cryptophycin 52, CPI-613, CTP-37, CTLA-4 monoclonal antibody (e.g., ipilimumab), CP-461, crizotinib (crizotinib), CV-247, cyanomorpholinodoxorubicin, cytarabine, D24851, dasatinib (dasatinib), decitabine (decitabine), dirithromycin (deoxorubicin), deoxydaunorubicin (deoxyrubicin), deoxyfumycin (deoxyoformycin), depsipeptide (depsipeptide), deoxyepothilone B, dexamethasone (dexamethasone), dexrazoxane (dexrazoxane), diethylstilbestrol (diethylsulbactam), deflazalidoxime (fludizolide), doxorazine (DMT-12), doxorazine (DMT-7410 DS, DMT-741, DMT-7410, DTE-1, DMT-7410, DMT-D, DMT-3, and DMT, DMT-3, and DMT including DMT, and DMT, Etorin (etotreotide), efaproxil (efaproxiral), eflornithine (eflornithine), EGFR inhibitor, EKB-569, EKB-509, enzastarin (enzastaurin), elimurol (eliscalicolmol), elsamitrucin (elsamimitrucin), epothilone B, epratuzumab (epratuzumab), EPZ-004777, ER-86526, erlotinib (erlotinib), ET-18-OCH3, ethynylcytidine, ethinylestradiol, irinotecan (exatecan), elsamitan mesylate, exemestane, esisultrin (exisultinde), fenretinide (fenretimide), phenanthroizumab (giumumab), floxuridine (floxuridine), folic acid glycoprotein (Acid), FOLFOX, IRLACTURETIN 4, FOLII (FOLIFOLITUM), FOLITUMOVITAMIN (FOLITUMOX), zotocin (FOLITUMORTA), FOLITUMORTIN (FOLITUMORTIN), FOLITUMORTA (FOLIN), FOLITUMORTIN (FOLITUMORTIN), FOLIN (FOLIT), FOLITUMIN (FOLIT), FOLIT (FOLIT-A), FOOTAMOTAMOTAMOTA), FOLIT-A (FOLIT-A), FOLIT-A (FOOTA), FOLIT-A (FOLIT-A), FOLIT-A (FOLIT-A), FOLIT-TAMIN-E (FOLIT-A), FOLIT-TAI), FOLIT-TAMIN-E (FOLIT-TAI), FOLIT-TAI (FOLIT-TAI), FOLIT-TAI (FOLIT-TAI, FOLIT-TAI (FOTAI, FOLIT-TAI, FOTAI, E (FOLIT-TAI, FOTAI, FOLIT-TAI, FOLITAI, FOLIT-TAI, FOLI, Gimatecan, glufosfamide, GCS-IOO, GDC-0623, GDC-0941 (pictelisib), GDC-0980, GDC-0032, GDC-0068, GDC-0349, GDC-0879, G17DT immunogen, GMK, GMX-1778, GPX-100, gp 100-peptide vaccine, GSK-5126766, GSK-690693, GSK-1120212 (trametinib)), GSK-1995010, GSK-2118436 (dabrafenib), GSK-2126458, GSK-2132231A, GSK-2334470, GSK-2110183, GSK-2141795, GSK-2636771, GSK-525762A/I-BET-762, 2016, and granisetron (G)ranitidine), herceptin (herceptin), hexamethylmelamine (hexamethylmelamine), histamine (histamine), homoharringtonine (homoharringtonine), hyaluronic acid (hyaluronic acid), hydroxyurea, progesterone hydroxycapronate, HDM-201, ibandronic acid (ibandronate), ibritumomab (ibritumomab), ibrutinib/PCI-32765, isatoilin (idasutilin), idarubinate (idatrexate), idarasib/101, estrenol (calenesterol), IDN-5109, IGF-1R inhibitors, IMC-1C11, IMC-A12 (sixutumumab), cixurnol (imunol), Indrilulan (INS), interferon alpha-2A, interferon alpha-2-1112, interferon alpha-2, interleukin alpha-2, interferon alpha-2-1117, interleukin alpha-2, interferon alpha-2-1117, interleukin alpha-2, interferon alpha-2, irninb, irninx, indone, irkum, indomethacin, indometha, Isopropyl platinum (iproplatin), iloprost (irofluven), isohalomethocholic acid-B (isohalophthalrin-B), isoflavone (isoflazone), isotretinoin (isotretinoin), ixabepilone (ixabepilone), JRX-2, JF-154, JQ-1, J-107088, conjugated estrogens, carhalide F (kahalid F), ketoconazole (ketoconazole), KW-2170, KW-2450, KU-55933, LCL-161, lobaplatin (lobaplatin), leflunomide (leflunomide), lenalidomide (lenalidomide), lenalidomide (lenograstin), LG (leuprolrolide), leuprolide (leuprolide), lisone (lisoprotein (lipin), leuprolide-d-651), leuprolide (lonamide (lonolide), valtretinone (loxacin), nevirapine (loxacin), linamin (loxacin), linagliptin (loxacin), linarin (loxacin), linamin (loxacin), linagliptin (loxacin), linagliptin (loxacin), linagliptin (loxacin), linagliptin (loxacin), linagliptin (loxacin), linalool), linagliptin (loxacin), linagliptin), linalool), linagliptin (loxacin), linalool), linagliptin (loxacin), linalool (loxacin), linalool (loxacin), linalool), linagliptin (loxacin), linalool (loxacin), linalool (loxacin), linalool (loxacin (lox, LY-S6AKT1, LY-2780301, LY-2109761/Caninidinib (galinisertib), malafosfamide (mafosfamide), marimastat (marimastat), mariroc (masoprocol), mechlorethamine (mechloroethamine), MEK inhibitors, MEK-162, methyltestosterone (methlesteron), methylprednisolone (methylprednisolone), MEDI-573, MEN-10755, MDX-H210, MDX-1379, MGV, midostaurin (midostaurin), minodronic acid (minodronic acid), mitomycin (mitomycin), mivoboolean (mivobu), MK-2206, MK-0646 (dalotuzumab)), MLN518, MLN-248, MLN-2480, MLN-01201280, mexofenamic acid (mivomycin)), MLN518, MLN-50, MLN-01255Gadolinium (Gd)(motexafin gadolinium)、MS-209, MS-275, MX6, neuraminic acid ester (neridronate), latinib (neratinib), sorafenib (Nexavar), simvastatin (neovastat), nilotinib (nilotinib), nimesulide (nimesulide), nitroglycerin (nitroglycerin), nolatrexed (nolatrexed), norrelin (norelin), N-acetylcysteine, NU-744106-benzylguanine, orlistat (oblimersen), omeprazole (omeprazole), olaparib (olaparib), bacteriophage (oncophage), oncoVEX (oncoVEX)^GM-CSFOrmiplatin (ormiplatin), paclitaxel otaxel (ortataxel), OX44 antibody, OSI-027, OSI-906 (lincetitinib), 4-1BB antibody, oxazostazole (oxantazole), estrogen (oestrogen), onapristone (onapristone), palbociclib (palbociclib)/PD-0332991, panitumumab (panitumumab), panobinostat, paltupirone (palozolone), pazopanib (pazopanib), pirfilgrastim (pegfilgrastim), PCK-3145, pirfenitin, PBI-1402, PBI-05204, PD0325901, PD-1 and PD-L1 antibodies (e.g. pemolizumab (perlizumab), nimbranmumab (PD), piriluzumab (Pildipidizumab), albumin (PD-47616), paclitaxel-stabilized taxol/paclitaxel (PEITVAT-7446), paclitaxel-7446 (PETU-D-7446), paclitaxel/PETU-D-7446, and so on, PF-05197281, PF-05212384, PF-04691502, PF-3758309, PHA-665752, PHT-427, P-04, PKC412, P54, PI-88, pelitinib (pelitinib), pemetrexed (pemetrexed), pantopraziquantel (pentrix), perifosine (perifosine), perillylalcohol (perlylcohol), pertuzumab (pertuzumab), pevonidat (pevon disttat), PI3K inhibitor, PI3K/mTOR inhibitor, PG-TXL, PG2, PLX-4032/RO-5185426 (Verofibrib)), PLX-3603/RO-5212054, PT-100, PWT-33597, PX-866, picoplatin (picoplatin), pivaloyloxymethylbutyrate (valoyloxymethyl butyrate), phenoxytryptophane (pkoxyphenol), polypeptine (pkpiroctone), polyprenedione (pkopicolide), polyprenolide (polyprenolide), polyprenolide (PKI (pexol, polyprenedione, polyprenolide), polyprenolide, PPT-3, PPT-TXylol, PPT-3-TXO, PPY (Perkininolate, PPY (PPY, porfinomycin (porfiromycin), posaconazole (posaconazole), prednisone (prednisone), prednisolone (prednisone), PRT-062607, quinamamide (quinamed), quinupristin (quinupristin), quinzatinib/AC 220, R115777, RAF-265, ramosetron (ramosetron), orchienase (rapiranase), RDEA-119/BAY 869766, RDEA-436, a butterfly-mycin (rebeccamycin) analog, a Receptor Tyrosine Kinase (RTK) inhibitor, thalidomide derivative (revimid), RG-7167, RG-7112, RG-7304, RG-7421, RG-7321, RG-7356, RG 7440, RG-7775, rhizoxin (rhizoxin), rhu-MAb, regexilin hydrochloride (rigosetiboate), risedronate (edrisinate), rituximab (robustamumab), robamumab (robamumab), rofecoxib (rofecoxib), romidepsin (romidepsin), RO-4929097, RO-31-7453, RO-5126766, RO-50760, RPR A, biuret (Robert-R), bixin (R-R), flurbiprofen (R-881), irinotecan (RX-R), bixin (R), and ryegrapridine (R) derivatives (rexate), RG-7167, RG-7112, RG-7312, RG-7321, RG-D, RG-D, RG-D, R-D, R (Rb, R-D, R (R-D, R (R-D, R-D, Ruxolitinib (ruxolitinib), S-9788, sabadicin (sabarubicin), SAHA, sabatabine, SAR-405838, sargrastim (sargramostim), satraplatin (satraplatin), SB-408075, SB-431542, Se-015/Ve-015, SU5416, SU6668, SDX-101, sellingo (selnexor), semastin (semustin), seocalcitol (seocalcitol), SM-11355, SN-38, SN-4071, SR-27897, SR-31747, SR-13668, SRL-172, sorafenib (sorafenib), spiroplatin (spirosplastin), squalamine (slaquamine), STF-31, anilino hydroxamic acid (sultialcine acetate), S-9788, Tactafine (Tactalidine), Tactalidine (TArtafin), Tactafine (Tactafine), Tactafine) (Tactafine), Tactafine (Tactafine) and Tactafine (Tactafine) L-32, Tactafine (Tactafine) and its (Tactafine ) can be used in, Tactafine (Tactafine, Takava, Tagetin, and its, Tagetin, and its, Tagetin, Tasolam (taurosulam), taxotere (taxotere), taxol (taxoprexin), tazarotene (tazarotene), tegafur (tegafur), temozolomide (temozolomide), timinil (tesilifene), testosterone (tetasterone), testosterone propionate, temini, tetraplatin (tetraplatin), tetrodotoxin (tetradotoxin), tizacitabine (tezacitabine), thalidomide (thiaidomide), tylunolide (theralux), terlopixin (thermiubin), thymopentin (thymofasin), thymosin (thymopectoracin), thiazolofurarin (tiazofurin), tipifarnib (tipifarnib), tirapazamine (tiapapamine), tolacacine (tacrine), tetrodotoxin (toxin), eugenol (tysotolomide), trematostatin (midtedentin), tretinomycin (midtedin), tretinomycin (107), tretinomycin (vitamin E (vitamin D), tretinomycin (vitamin E (D), tretinomycin (vitamin E (D-D), tretinomycin (D (vitamin E), tretinomycin (D), tretinomycin (D-D), tretinomycin (D-D,Trastuzumab (traszuzumab), tremelimumab (tremelimumab), tretinoin (tretinoinin), triacetyluridine (triacetyluridine), triamcinolone (triapine), triciribine (triciribine), tremetrexate (trimetrexate), TLK-286TXD 258, tyloxa (tykerb)/lapatinib (tyverib), uracil (urocidin), valproic acid (valproic acid), valrubicin (valrubicin), vandetanib (vandetanib), vatalanib (vatalanib), vincristine, vinflunine (vinflunine), virulence factor (virulizin), virgule (vismodegib), vosaloxin, WX-UK1, WX-554, Viktisib (vinfluxib), XAV-9, peruviveleda (vilab), Vmeizumab-41147, ZX-XL-5923, ZDZX-XL-59XL-20, ZDrexXL-59xl, ZDrexate (tyx-5/XL-5), TXylvanib (tyne (tyx-9, trexab), trexabin (vinflunixin), and (vxl-35, XL-35, XL-9, XL-9, XL-XL (vxl-9, XL-9-XL-9, XL-9, XL-9, XL-9, trexab, XL-XL (valrubicin, trexab, trexa-XL-9, trexab, trexa, trexab, tre, ZD-4054, ZD-0473, ZD-6126, ZD-9331, ZDI839, ZSTK-474, sodium zoledronate (zoledronate), and zosuquidar.

In some embodiments, the combination therapy as described herein involves an LRP5 antagonist and an anti-PD-1 antibody as described herein without any additional chemotherapeutic agent.

Hyperproliferative diseases/cancers

The combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are useful in the treatment and/or prevention of hyperproliferative disorders, in particular cancer.

In certain embodiments, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are useful for treating hyperproliferative disorders, particularly cancer.

As used herein, "hyperproliferative diseases" refer to conditions in which cell growth is increased above normal levels. For example, hyperproliferative diseases or disorders include malignant diseases (e.g., esophageal cancer, colon cancer, biliary tract cancer) and non-malignant diseases (e.g., atherosclerosis, benign hyperplasia, benign prostatic hypertrophy).

In a preferred embodiment, the hyperproliferative disorder is cancer.

Cancer can be classified in two ways: depending on the tissue type (histological type) in which the cancer originates and on the major site or location in the body in which the cancer first develops. The most common sites of cancer development include the skin, lung, breast, prostate, colon and rectum, cervix and uterus, and blood chambers.

The combinations, compositions, kits, uses, methods and compounds (including all examples) used according to the invention may be useful in the treatment of various hyperproliferative disorders, particularly cancer, including (for example, but not limited to) the following:

gastrointestinal tract cancer, such as esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular cancer, biliary tract cancer (e.g., biliary duct cancer), gallbladder cancer, pancreatic cancer, or colorectal cancer (CRC);

melanoma;

bladder cancer; and

lung cancer (e.g., NSCLC).

In some embodiments of the invention, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are for the treatment of gastrointestinal cancer, preferably esophageal cancer (e.g., gastroesophageal junction cancer), gastric (stomach) cancer, hepatocellular cancer, biliary tract cancer (e.g., cholangiocarcinoma), gallbladder cancer, pancreatic cancer or colorectal cancer (CRC). Especially preferred for the treatment of these cancers are LRP5#2 as LRP5 antagonist and PD1-1 as anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In some embodiments of the invention, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) used according to the invention are for the treatment of melanoma. It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In some embodiments of the invention, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) used according to the invention are for the treatment of bladder cancer. It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In some embodiments of the invention, the combinations, compositions, kits, uses, methods and compounds, including all embodiments, used according to the invention are for the treatment of lung cancer, such as non-small cell lung cancer (NSCLC). It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In another embodiment of the invention, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are used for the treatment of a cancer patient (e.g. a patient suffering from (i) a cancer of the gastrointestinal tract, e.g. of the esophagus, stomach, hepatocellular, biliary tract, pancreas or colorectal, (ii) melanoma, (iii) bladder or (iv) lung) who is not undergoing treatment with a checkpoint inhibitor or immunomodulator, i.e. e.g. a patient who is not undergoing treatment with an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In another embodiment of the invention, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are used for the treatment of a cancer patient (e.g. a patient suffering from (i) a gastrointestinal cancer, e.g. esophageal cancer, gastric cancer, hepatocellular cancer, biliary tract cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer) who relapses during, subsequent to or after treatment with a checkpoint inhibitor or immunomodulator, i.e. e. e.g. a patient who relapses during, subsequent to or after treatment with a PD-1 antagonist (e.g. an anti-PD-1 antibody). It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

Therapeutic applicability of the combination therapy according to the invention may include first-line, second-line, third-line or other line treatment of a patient (e.g., a patient suffering from (i) gastrointestinal cancer such as esophageal cancer, gastric cancer, hepatocellular cancer, biliary tract cancer, gallbladder cancer, pancreatic cancer, or colorectal cancer, (ii) melanoma, (iii) bladder cancer, or (iv) lung cancer). The cancer may be metastatic, periodic, recurrent, resistant or refractory to one or more anticancer treatments. Thus, these patients may not be treated or may have received one or more prior anti-cancer therapies that have not yet completely cured the disease.

Patients who relapse and/or are resistant to one or more anti-cancer agents (e.g. a single component of a combination, or a standard chemotherapeutic agent) are also amenable to combination therapy according to the invention, e.g. to two-or three-line treatment cycles (optionally further combined with one or more other anti-cancer agents), e.g. as an additional combination or as a replacement therapy.

Thus, some of the combination therapies of the invention disclosed herein are effective in treating individuals (e.g., patients suffering from (i) gastrointestinal cancer, e.g., esophageal, gastric, hepatocellular, biliary, pancreatic, or colorectal cancer, (ii) melanoma, (iii) bladder, or (iv) lung cancer), whose cancer has relapsed, or whose cancer has become drug-resistant or multi-drug resistant, or for whom monotherapy using one or more anticancer agents (e.g., a single component in combination, or a standard chemotherapeutic agent) or for whom first, second, or more lines of combination therapy have failed.

A cancer that has an initial response to an anticancer drug may recur and when the anticancer drug is no longer effective to treat an individual with the cancer, the cancer may become resistant to the anticancer drug, e.g., even if an increased dose of the anticancer drug is administered. Cancers that have developed resistance to two or more anticancer drugs are referred to as multidrug resistant.

In preferred embodiments, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) used according to the invention are used for treating a cancer patient (e.g., a patient suffering from (i) gastrointestinal cancer, e.g., esophageal cancer, gastric cancer, hepatocellular cancer, biliary tract cancer, pancreatic cancer, or colorectal cancer, (ii) melanoma, (iii) bladder cancer, or (iv) lung cancer) that has not previously been treated with one or more immune checkpoint inhibitors and/or immune modulators, e.g., one or more PD-1 antagonists (e.g., anti-PD 1 antibodies). It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In another preferred embodiment, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are used for treating a cancer patient (e.g., a patient suffering from (i) gastrointestinal cancer, e.g., esophageal cancer, gastric cancer, hepatocellular cancer, biliary tract cancer, gallbladder cancer, pancreatic cancer or colorectal cancer, (ii) melanoma, (iii) bladder cancer or (iv) lung cancer) that is refractory or resistant to checkpoint inhibitor therapy (e.g., to treatment with one or more immune checkpoint inhibitors and/or immune modulators, e.g., one or more PD-1 antagonists (e.g., anti-PD 1 antibodies)). It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

In an alternative preferred embodiment, the combinations, compositions, kits, uses, methods and compounds (including all embodiments) for use according to the invention are used for treating a cancer patient suffering from any solid tumor that is refractory or resistant to checkpoint inhibitor therapy, e.g. to treatment with one or more immune checkpoint inhibitors and/or immune modulators, e.g. one or more PD-1 antagonists, e.g. an anti-PD 1 antibody. It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Examples of solid tumors are well known in the art. Likewise, the terms refractory or resistant are also known to those skilled in the art and are used in the present application according to the definitions adopted in the art.

Tumors that are refractory or resistant to checkpoint inhibitor therapy are also referred to herein as "anti-immunotherapeutic tumors" or "anti-immunotherapeutic non-T cell inflammatory tumors". It has recently been found that in the microenvironment of many tumors, high expression of specific immune cells can be found. This is referred to in the art as the "T-cell inflamed phenotype" and this phenotype has been observed to be associated with these tumors amenable to treatment using multiple immunotherapies including therapeutic vaccines and checkpoint blocking antibodies (e.g., anti-PD-1 antibodies). On the other hand, some tumors lack this expression of immune cells in their microenvironment. These tumors are known in the art as "non-T cell inflammatory tumors" and are found to lack clinical benefit to immunotherapy, particularly with anti-PD-1 antibodies. According to the present invention, the latter type of tumor with active Wnt signaling is a preferred target for the combination therapy claimed in the present invention. It is especially preferred to treat these cancers with LRP5#2 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#6 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. It is especially preferred to treat these cancers with LRP5#7 as an LRP5 antagonist and PD1-1 as an anti-PD-1 antibody. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-2 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#2 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#6 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers. Especially preferred are LRP5#7 as LRP5 antagonist and PD1-3 as anti-PD-1 antibody for the treatment of these cancers.

The present invention is not to be limited in scope by the specific embodiments described herein. Various modifications of the invention, in addition to those described herein, will be apparent to those skilled in the art from the disclosure of the invention. Such modifications are intended to fall within the scope of the appended claims.

All patent applications cited in this application are incorporated by reference in their entirety.

Example 1

Exemplary LRP5 antagonist combination with mouse PD-1 antibody anti-tumor Activity in a subcutaneous syngeneic mouse model derived from the breast cancer cell line EMT6 in Balb/c mice

An exemplary LRP5 antagonist was tested for utility as a single agent and in combination with a mouse PD-1 antibody in an s.c. cell line-derived mouse breast cancer (EMT6) isogenic model.

BALB/cJBomTac mice were used in this study. Each mouse was injected with 1x10⁶EMT6 breast cancer cells to establish tumors. Tumor volume was measured at least three times per week using calipers. When the tumor has reached about 120mm³Start treatment and end after 30 days.

Ten tumor-bearing animals were treated with an exemplary LRP5 antagonist, i.v., i.p. anti-PD-1 antibody, or a combination of two compounds, intravenously (i.v.) twice weekly and twice weekly. Ten animals were used in the vehicle/isotype control-treated group. For moral reasons based on tumor masses (tumors ≥ 1.5 cm)³) Animals were euthanized at the end of the study.

Cells

EMT6 cells were obtained from ATCC (Cat. No.)CRL2755^TM). Establishing a Master Cell Bank (MCB) and a Working Cell Bank (WCB). At 37 ℃ and 5% CO₂The cells were then cultured in T175 tissue culture flasks. The culture medium is 15% fetal calf serum (Fetal bone Serum Characterized; directory number SH 30071.03; supplied by Thermo Scientific), and 2mM L-glutamine (L-glutamine 200mM (100X); reference 25030-; supplied by Gibco by Life Technologies) supplemented wecker MB 752/1. Cultures were isolated every two to three days at a ratio of 1:10/1: 15.

Mouse

Mice were 7 to 8 week old BALB/cJBomTac purchased from Taconic, Denmark. Upon arrival at the animal facility, the mice were allowed to acclimate to ambient conditions for at least 5 days before being used in the experiment. Ten of these mice were housed in each group at 21.5. + -. 1.5 ℃ and 55. + -. 10% humidity under standardized conditionsType III cages. Standardized irradiated diet (PROVIMI KLIBA) and autoclaved tap water were provided ad libitum. Microchips implanted subcutaneously under isoflurane anesthesia were used to identify individual mice. The caged card displays study number, animal number, compound and dose, route of administration and schedule maintained by these animals throughout the study.

Administration of test Compounds

The LRP5 antagonist was suspended in histidine buffer pH 6.5 and administered twice weekly in an administration volume of 10mL/kg at a dose of 15mg/kg for the first two weeks.

PD-1 antibody was diluted in PBS and injected intraperitoneally twice weekly at a dose of 10mg/kg in a volume of 10mL/kg per mouse until the end of the study.

Monitoring tumor growth and disease progression

Tumor diameters were measured with calipers three times per week (monday, wednesday and friday). Volume of each tumor [ in mm ]³Meter]Calculated according to the formula "tumor volume length diameter 2 pi/6". To monitor the side effects of treatment, mice were examined daily for abnormalities and body weights were measured daily. Animals were sacrificed at the end of the study. Will have necrotic tumors or tumor sizes exceeding 1500mm for moral reasons during the study³The animals were sacrificed as early as possible.

Results

Treatment of ETM6 tumors with mouse antibodies against PD-1 or LRP5 antagonists resulted in moderate tumor growth inhibition. The combination of the LRP5 antagonist and the PD-1 antibody resulted in a significantly increased efficacy when compared to single agent administration, which caused tumor regression in 5 of 10 mice when tumor regression was observed in a lesser number of mice when compared to monotherapy (tumor regression was only observed in 1 and 3 of 10 mice treated with single agent PD-1 antibody or LRP5 antagonist, respectively). The results demonstrating the synergistic effect of the combination administration compared to the monotherapy are shown in figure 1. Furthermore, the combination of LRP5 antagonist and PD-1 antibody increased in table 5 as from the start of treatment to when the tumor was treated when compared to monotherapyThe volume of the product is at least 500mm³Time of day intervals increased survival was recorded.

Table 5 shows the anti-tumor activity of LRP5 antagonists as single agents and in combination with mouse PD-1 antibodies. Median means from the start of treatment to when the tumor volume reached at least 500mm³The time interval (days).

TABLE 5

Sequence listing

<110> Boringer Invighan International Ltd

<120> anti-cancer combination therapy

<130> 12-0438-WO-1

<160> 62

<170> BiSSAP 1.3.6

<210> 1

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1HCDR1

<400> 1

Gly Phe Thr Phe Ser Ala Ser Ala Met Ser

1 5 10

<210> 2

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1HCDR2

<400> 2

Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val Lys

1 5 10 15

Gly

<210> 3

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1HCDR3

<400> 3

His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr

1 5 10

<210> 4

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1LCDR1

<400> 4

Arg Ala Ser Glu Asn Ile Asp Thr Ser Gly Ile Ser Phe Met Asn

1 5 10 15

<210> 5

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1LCDR2

<400> 5

Val Ala Ser Asn Gln Gly Ser

1 5

<210> 6

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-1LCDR3

<400> 6

Gln Gln Ser Lys Glu Val Pro Trp Thr

1 5

<210> 7

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2HCDR1

<400> 7

Gly Phe Thr Phe Ser Ala Ser Ala Met Ser

1 5 10

<210> 8

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2HCDR2

<400> 8

Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val Lys

1 5 10 15

Gly

<210> 9

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2HCDR3

<400> 9

His Ser Asn Pro Asn Tyr Tyr Ala Met Asp Tyr

1 5 10

<210> 10

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2LCDR1

<400> 10

Arg Ala Ser Glu Asn Ile Asp Thr Ser Gly Ile Ser Phe Met Asn

1 5 10 15

<210> 11

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2LCDR2

<400> 11

Val Ala Ser Asn Gln Gly Ser

1 5

<210> 12

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-2LCDR3

<400> 12

Gln Gln Ser Lys Glu Val Pro Trp Thr

1 5

<210> 13

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3HCDR1

<400> 13

Gly Phe Thr Phe Ser Lys Ser Ala Met Ser

1 5 10

<210> 14

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3HCDR2

<400> 14

Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val Lys

1 5 10 15

Gly

<210> 15

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3HCDR3

<400> 15

His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr

1 5 10

<210> 16

<211> 15

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3LCDR1

<400> 16

Arg Ala Ser Glu Asn Ile Asp Val Ser Gly Ile Ser Phe Met Asn

1 5 10 15

<210> 17

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3LCDR2

<400> 17

Val Ala Ser Asn Gln Gly Ser

1 5

<210> 18

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> PD1-3LCDR3

<400> 18

Gln Gln Ser Lys Glu Val Pro Trp Thr

1 5

<210> 19

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VH1

<400> 19

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ala Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 20

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VL1

<400> 20

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Thr Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 21

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VH2

<400> 21

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ala Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Pro Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 22

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VL2

<400> 22

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Thr Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 23

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VH3

<400> 23

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 24

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VL3

<400> 24

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 25

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VH4

<400> 25

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 26

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VL4

<400> 26

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 27

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VH5

<400> 27

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 28

<211> 111

<212> PRT

<213> Artificial sequence

<220>

<223> PD1VL5

<400> 28

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys

100 105 110

<210> 29

<211> 446

<212> PRT

<213> Artificial sequence

<220>

<223> PD1HC1

<400> 29

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ala Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

<210> 30

<211> 218

<212> PRT

<213> Artificial sequence

<220>

<223> PD1LC1

<400> 30

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Thr Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 31

<211> 446

<212> PRT

<213> Artificial sequence

<220>

<223> PD1HC2

<400> 31

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ala Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Pro Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

<210> 32

<211> 218

<212> PRT

<213> Artificial sequence

<220>

<223> PD1LC2

<400> 32

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Thr Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 33

<211> 446

<212> PRT

<213> Artificial sequence

<220>

<223> PD1HC3

<400> 33

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

<210> 34

<211> 218

<212> PRT

<213> Artificial sequence

<220>

<223> PD1LC3

<400> 34

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 35

<211> 446

<212> PRT

<213> Artificial sequence

<220>

<223> PD1HC4

<400> 35

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

<210> 36

<211> 218

<212> PRT

<213> Artificial sequence

<220>

<223> PD1LC4

<400> 36

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 37

<211> 446

<212> PRT

<213> Artificial sequence

<220>

<223> PD1HC5

<400> 37

Glu Val Met Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Lys Ser

20 25 30

Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Ser Gly Gly Gly Gly Asp Thr Tyr Tyr Ser Ser Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg His Ser Asn Val Asn Tyr Tyr Ala Met Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val

115 120 125

Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala

130 135 140

Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser

145 150 155 160

Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val

165 170 175

Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro

180 185 190

Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys

195 200 205

Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro

210 215 220

Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val

225 230 235 240

Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr

245 250 255

Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu

260 265 270

Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys

275 280 285

Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser

290 295 300

Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys

305 310 315 320

Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile

325 330 335

Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro

340 345 350

Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu

355 360 365

Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn

370 375 380

Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser

385 390 395 400

Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg

405 410 415

Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu

420 425 430

His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly

435 440 445

<210> 38

<211> 218

<212> PRT

<213> Artificial sequence

<220>

<223> PD1LC5

<400> 38

Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly

1 5 10 15

Glu Arg Ala Thr Met Ser Cys Arg Ala Ser Glu Asn Ile Asp Val Ser

20 25 30

Gly Ile Ser Phe Met Asn Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro

35 40 45

Lys Leu Leu Ile Tyr Val Ala Ser Asn Gln Gly Ser Gly Ile Pro Ala

50 55 60

Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser

65 70 75 80

Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Ser Lys

85 90 95

Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg

100 105 110

Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln

115 120 125

Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr

130 135 140

Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser

145 150 155 160

Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr

165 170 175

Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys

180 185 190

His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro

195 200 205

Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215

<210> 39

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> motif for B7-1 and B7-2 binding

<400> 39

Met Tyr Pro Pro Tyr

1 5

<210> 40

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> F0129097A08 CDR1

<400> 40

Thr Tyr Val Met Gly

1 5

<210> 41

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> F0129097A08 CDR2

<400> 41

Ala Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 42

<211> 18

<212> PRT

<213> Artificial sequence

<220>

<223> F0129097A08 CDR3

<400> 42

Ser Arg Gly Thr Ser Thr Pro Ser Arg Ala Ser Gly Val Ser Arg Tyr

1 5 10 15

Asp Tyr

<210> 43

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> F0129093A02 CDR1

<400> 43

Arg Tyr Ala Val Ala

1 5

<210> 44

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> F0129093A02 CDR2

<400> 44

Ala Ile Thr Trp Ser Ser Gly Arg Ile Asp Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 45

<211> 20

<212> PRT

<213> Artificial sequence

<220>

<223> F0129093A02 CDR3

<400> 45

Asp Arg Arg Pro Arg Ser Thr Gly Arg Ser Gly Thr Gly Ser Pro Ser

1 5 10 15

Thr Tyr Asp Tyr

<210> 46

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> F012904 6C10(E1A,N32G) CDR1

<400> 46

Ile Gly Ala Met Gly

1 5

<210> 47

<211> 16

<212> PRT

<213> Artificial sequence

<220>

<223> F012904 6C10(E1A,N32G) CDR2 / F012904 6C10 CDR2

<400> 47

Ala Val Ser Ser Gly Gly Ser Thr Tyr Tyr Val Asp Ser Val Lys Gly

1 5 10 15

<210> 48

<211> 12

<212> PRT

<213> Artificial sequence

<220>

<223> F012904 6C10(E1A,N32G) CDR3 / F012904 6C10 CDR3

<400> 48

Glu Thr Gly Pro Tyr Gly Pro Pro Lys Arg Asp Tyr

1 5 10

<210> 49

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> F012904 6C10 CDR1

<400> 49

Ile Asn Ala Met Gly

1 5

<210> 50

<211> 127

<212> PRT

<213> Artificial sequence

<220>

<223> F0129097A08(E1A,V23A) VHH

<400> 50

Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr

20 25 30

Val Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ala Ser Arg Gly Thr Ser Thr Pro Ser Arg Ala Ser Gly Val Ser

100 105 110

Arg Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

<210> 51

<211> 130

<212> PRT

<213> Artificial sequence

<220>

<223> F0129093A02 VHH

<400> 51

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Arg Tyr

20 25 30

Ala Val Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Thr Trp Ser Ser Gly Arg Ile Asp Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ala Asp Arg Arg Pro Arg Ser Thr Gly Arg Ser Gly Thr Gly Ser

100 105 110

Pro Ser Thr Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser

115 120 125

Ser Ala

130

<210> 52

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> F0129046C10(E1A,N32G) VHH

<400> 52

Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Arg Ile Gly

20 25 30

Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ala Val Ser Ser Gly Gly Ser Thr Tyr Tyr Val Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Arg Glu Thr Gly Pro Tyr Gly Pro Pro Lys Arg Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 53

<211> 120

<212> PRT

<213> Artificial sequence

<220>

<223> F0129046C10 VHH

<400> 53

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Arg Ile Asn

20 25 30

Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ala Val Ser Ser Gly Gly Ser Thr Tyr Tyr Val Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Arg Glu Thr Gly Pro Tyr Gly Pro Pro Lys Arg Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser

115 120

<210> 54

<211> 5

<212> PRT

<213> Artificial sequence

<220>

<223> Alb11 Domain CDR1

<400> 54

Ser Phe Gly Met Ser

1 5

<210> 55

<211> 17

<212> PRT

<213> Artificial sequence

<220>

<223> Alb11 Domain CDR2

<400> 55

Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp Ser Val Lys

1 5 10 15

Gly

<210> 56

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> Alb11 Domain CDR3

<400> 56

Gly Gly Ser Leu Ser Arg

1 5

<210> 57

<211> 432

<212> PRT

<213> Artificial sequence

<220>

<223> polypeptide capable of specifically binding to LRP5 (1)

<400> 57

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Arg Thr Phe Ser Thr Tyr

20 25 30

Val Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ala Ser Arg Gly Thr Ser Thr Pro Ser Arg Ala Ser Gly Val Ser

100 105 110

Arg Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

130 135 140

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

165 170 175

Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

180 185 190

Ser Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

195 200 205

Trp Val Ser Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp

210 215 220

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr

225 230 235 240

Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr

245 250 255

Tyr Cys Thr Ile Gly Gly Ser Leu Ser Arg Ser Ser Gln Gly Thr Leu

260 265 270

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

275 280 285

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

290 295 300

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

305 310 315 320

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

325 330 335

Ser Gly Ser Ile Phe Arg Ile Asn Ala Met Gly Trp Tyr Arg Gln Ala

340 345 350

Pro Gly Lys Gln Arg Glu Leu Val Ala Ala Val Ser Ser Gly Gly Ser

355 360 365

Thr Tyr Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

370 375 380

Asn Ser Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu

385 390 395 400

Asp Thr Ala Val Tyr Tyr Cys Asn Arg Glu Thr Gly Pro Tyr Gly Pro

405 410 415

Pro Lys Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

420 425 430

<210> 58

<211> 435

<212> PRT

<213> Artificial sequence

<220>

<223> polypeptide capable of specifically binding to LRP5 (2)

<400> 58

Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Arg Ile Gly

20 25 30

Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ala Val Ser Ser Gly Gly Ser Thr Tyr Tyr Val Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Arg Glu Thr Gly Pro Tyr Gly Pro Pro Lys Arg Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly

115 120 125

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

130 135 140

Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val

145 150 155 160

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn Ser Leu Arg Leu Ser

165 170 175

Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe Gly Met Ser Trp Val

180 185 190

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Ser Ile Ser Gly

195 200 205

Ser Gly Ser Asp Thr Leu Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr

210 215 220

Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr Leu Gln Met Asn Ser

225 230 235 240

Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Thr Ile Gly Gly Ser

245 250 255

Leu Ser Arg Ser Ser Gln Gly Thr Leu Val Thr Val Ser Ser Gly Gly

260 265 270

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

275 280 285

Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly

290 295 300

Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly

305 310 315 320

Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Phe Ser Arg

325 330 335

Tyr Ala Val Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe

340 345 350

Val Ala Ala Ile Thr Trp Ser Ser Gly Arg Ile Asp Tyr Ala Asp Ser

355 360 365

Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val

370 375 380

Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr

385 390 395 400

Cys Ala Ala Asp Arg Arg Pro Arg Ser Thr Gly Arg Ser Gly Thr Gly

405 410 415

Ser Pro Ser Thr Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val

420 425 430

Ser Ser Ala

435

<210> 59

<211> 433

<212> PRT

<213> Artificial sequence

<220>

<223> polypeptide capable of specifically binding to LRP5 (3)

<400> 59

Ala Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr

20 25 30

Val Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ala Ser Arg Gly Thr Ser Thr Pro Ser Arg Ala Ser Gly Val Ser

100 105 110

Arg Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Gly

115 120 125

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

130 135 140

Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly

145 150 155 160

Gly Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro

165 170 175

Gly Asn Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser

180 185 190

Ser Phe Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu

195 200 205

Trp Val Ser Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp

210 215 220

Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr

225 230 235 240

Leu Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr

245 250 255

Tyr Cys Thr Ile Gly Gly Ser Leu Ser Arg Ser Ser Gln Gly Thr Leu

260 265 270

Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly

275 280 285

Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly

290 295 300

Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Val Glu Ser Gly

305 310 315 320

Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala

325 330 335

Ser Gly Ser Ile Phe Arg Ile Gly Ala Met Gly Trp Tyr Arg Gln Ala

340 345 350

Pro Gly Lys Gln Arg Glu Leu Val Ala Ala Val Ser Ser Gly Gly Ser

355 360 365

Thr Tyr Tyr Val Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp

370 375 380

Asn Ser Lys Asn Thr Val Tyr Leu Gln Met Asn Ser Leu Arg Pro Glu

385 390 395 400

Asp Thr Ala Val Tyr Tyr Cys Asn Arg Glu Thr Gly Pro Tyr Gly Pro

405 410 415

Pro Lys Arg Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

420 425 430

Ala

<210> 60

<211> 115

<212> PRT

<213> Artificial sequence

<220>

<223> Alb11 VHH

<400> 60

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Asn

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Phe

20 25 30

Gly Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ser Ser Ile Ser Gly Ser Gly Ser Asp Thr Leu Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Thr Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Thr Ile Gly Gly Ser Leu Ser Arg Ser Ser Gln Gly Thr Leu Val Thr

100 105 110

Val Ser Ser

115

<210> 61

<211> 121

<212> PRT

<213> Artificial sequence

<220>

<223> F0129046C10(N32G) VHH

<400> 61

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Ile Phe Arg Ile Gly

20 25 30

Ala Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gln Arg Glu Leu Val

35 40 45

Ala Ala Val Ser Ser Gly Gly Ser Thr Tyr Tyr Val Asp Ser Val Lys

50 55 60

Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr Leu

65 70 75 80

Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn

85 90 95

Arg Glu Thr Gly Pro Tyr Gly Pro Pro Lys Arg Asp Tyr Trp Gly Gln

100 105 110

Gly Thr Leu Val Thr Val Ser Ser Ala

115 120

<210> 62

<211> 127

<212> PRT

<213> Artificial sequence

<220>

<223> F0129097A08 VHH

<400> 62

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Arg Thr Phe Ser Thr Tyr

20 25 30

Val Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val

35 40 45

Ala Ala Ile Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Ala Ser Arg Gly Thr Ser Thr Pro Ser Arg Ala Ser Gly Val Ser

100 105 110

Arg Tyr Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser

115 120 125

104页详细技术资料下载

上一篇：一种医用注射器针头装配设备

下一篇：作为SHP2拮抗剂的嘧啶酮衍生物

Anti-cancer combination therapy

相关技术

网友询问留言