JPWO2021119516A5

JPWO2021119516A5 -

Info

Publication number: JPWO2021119516A5
Application number: JP2022535520A
Authority: JP
Publication date: 2023-12-19

Claims

A physiologically active polypeptide drug construct comprising, from the N-terminus to the C-terminus (D1-D2-D3),
1) a D1 domain (“D1”) that is a tissue or diseased area target portion; 2) a D2 domain (“D2”) that is a physiologically active portion; and 3) a D3 domain (“D3”) that is a shielding portion. wherein D1 serves to target the bioactive moiety to the desired treatment site; D3 is capable of masking the functional activity of D2 until activated at the desired treatment site; or
1) a D1 domain (“D1”) that is a dual functional moiety; 2) a D2 domain (“D2”) that is a physiologically active moiety; and 3) a D3 domain (“D3”) that is a shielding moiety. D1 serves to target and retain the bioactive moiety at the desired treatment site; D3 is capable of masking the functional activity of D2 until it is activated at the desired treatment site. ;
D2 is an IL-15 variant polypeptide selected from the group of polypeptides having the amino acid sequences set forth in SEQ ID NOs: 182-192 and 199-215;
D1 is an Fc domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 156, and each amino acid sequence set forth in SEQ ID NO: 166 to 168;
Said D3 domain is a cognate receptor/binding partner for IL-15 (or a variant thereof) and comprises the amino acid sequence set forth in SEQ ID NO: 4 or any functional fragment thereof, or the amino acid sequence set forth in SEQ ID NO: 5. containing an array,
Bioactivated polypeptide drug construct.

A physiologically active polypeptide drug construct comprising, from the N-terminus to the C-terminus (D1-D2-D3),
1) a D1 domain (“D1”) that is a tissue or diseased area target portion; 2) a D2 domain (“D2”) that is a physiologically active portion; and 3) a D3 domain (“D3”) that is a shielding portion. wherein D1 serves to target the bioactive moiety to the desired treatment site; D3 is capable of masking the functional activity of D2 until activated at the desired treatment site; or
1) a D1 domain (“D1”) that is a dual functional moiety; 2) a D2 domain (“D2”) that is a physiologically active moiety; and 3) a D3 domain (“D3”) that is a shielding moiety. D1 serves to target and retain the bioactive moiety at the desired treatment site; D3 is capable of masking the functional activity of D2 until it is activated at the desired treatment site. ;
D2 is an IL-2 variant polypeptide selected from the group of polypeptides having the amino acid sequence set forth in SEQ ID NOs: 232-247;
D1 is an Fc domain having an amino acid sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 156, and each amino acid sequence set forth in SEQ ID NO: 166 to 168;
said D3 domain is a cognate receptor/binding partner for IL-2 (or a variant thereof) and comprises the amino acid sequence set forth in SEQ ID NO: 10 and 267-270;
Bioactivated polypeptide drug construct.

A physiologically active polypeptide drug construct comprising, from the N-terminus to the C-terminus (D1-D2-D3),
1) a D1 domain (“D1”) that is a tissue or diseased area target portion; 2) a D2 domain (“D2”) that is a physiologically active portion; and 3) a D3 domain (“D3”) that is a shielding portion. wherein D1 serves to target the bioactive moiety to the desired treatment site; D3 is capable of masking the functional activity of D2 until activated at the desired treatment site; or
1) a D1 domain (“D1”) that is a dual functional moiety; 2) a D2 domain (“D2”) that is a physiologically active moiety; and 3) a D3 domain (“D3”) that is a shielding moiety. D1 serves to target and retain the bioactive moiety at the desired treatment site; D3 is capable of masking the functional activity of D2 until it is activated at the desired treatment site. ;
D2 is an IL-15 variant polypeptide selected from the group of polypeptides having the amino acid sequences set forth in SEQ ID NOs: 182-192 and 199-215.
D1 is the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 195 and 196, the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 197 and 198, and the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 275 and 276. , or an antagonist humanized PD-1 antibody selected from antibodies comprising the heavy chain and light chain amino acid sequences set forth in SEQ ID NOs: 277 and 278,
Said D3 domain is a cognate receptor/binding partner for IL-15 (or a variant thereof) and comprises the amino acid sequence set forth in SEQ ID NO: 4 or any functional fragment thereof, or the amino acid sequence set forth in SEQ ID NO: 5. containing an array,
Bioactivated polypeptide drug construct.

A physiologically active polypeptide drug construct comprising, from the N-terminus to the C-terminus (D1-D2-D3),
1) a D1 domain (“D1”) that is a tissue or diseased area target portion; 2) a D2 domain (“D2”) that is a physiologically active portion; and 3) a D3 domain (“D3”) that is a shielding portion. wherein D1 serves to target the bioactive moiety to the desired treatment site; D3 is capable of masking the functional activity of D2 until activated at the desired treatment site; or
1) a D1 domain (“D1”) that is a dual functional moiety; 2) a D2 domain (“D2”) that is a physiologically active moiety; and 3) a D3 domain (“D3”) that is a shielding moiety. D1 serves to target and retain the bioactive moiety at the desired treatment site; D3 is capable of masking the functional activity of D2 until it is activated at the desired treatment site. ;
D2 is an IL-2 variant polypeptide selected from the group of polypeptides having the amino acid sequence set forth in SEQ ID NOs: 232-247;
D1 is the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 195 and 196, the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 197 and 198, and the heavy chain and light chain amino acid sequence set forth in SEQ ID NO: 275 and 276. , or an antagonist humanized PD-1 antibody selected from antibodies comprising the heavy chain and light chain amino acid sequences set forth in SEQ ID NOs: 277 and 278,
said D3 domain is a cognate receptor/binding partner for IL-2 (or a variant thereof) and comprises the amino acid sequence set forth in SEQ ID NO: 10 and 267-270;
Bioactivated polypeptide drug construct.

Claims that the D1, D2, and D3 domains of the construct are each in monomeric form, each in dimeric form, or in the form of a dimer and a monomer combined together. The construct according to any one of items 1 to 4 .

6. D2 is attached to D1 by a peptide linker ("L1") selected from the group consisting of a protease-cleavable peptide linker, and a non-cleavable peptide linker . construct.

7. The construct of claim 6 , wherein said protease cleavable peptide linker is selected from the group consisting of sequences set forth in SEQ ID NOs: 71-96, 157-161 and 298.

7. The construct of claim 6 , wherein the non-cleavable peptide linker is selected from the group consisting of the sequences set forth in SEQ ID NOs: 107-127.

6. D2 is attached to D3 by a peptide linker ("L2") selected from the group consisting of a protease-cleavable peptide linker, and a non-cleavable peptide linker . construct.

10. The construct of claim 9 , wherein said protease cleavable peptide linker is selected from the group consisting of sequences set forth in SEQ ID NOs: 71-96, 157-161 and 298.

10. The construct of claim 9 , wherein the non-cleavable peptide linker is selected from the group consisting of each sequence set forth in SEQ ID NOs: 107-127.

Both L1 and L2 are protease cleavable peptide linkers , or both L1 and L2 are non-cleavable peptide linkers, or L1 is a protease cleavable peptide linker and L2 is non-cleavable. The construct according to any one of claims 1 to 11 , wherein L1 is a non-cleavable peptide linker and L2 is a protease-cleavable peptide linker .

A pharmaceutical composition comprising a construct according to any one of claims 1 to 12 in admixture with a pharmaceutically acceptable carrier.

14. The pharmaceutical composition of claim 13 for use in treating cancer or cancer metastasis in a subject.

15. The pharmaceutical composition of claim 14 , wherein the treatment further comprises a second therapeutic agent or method capable of treating cancer or cancer metastasis in the subject.

A nucleic acid molecule encoding a construct according to any one of claims 1 to 12 .

An expression vector comprising the nucleic acid molecule according to claim 16 .

A host cell comprising the expression vector of claim 17 .

19. A method for producing the bioactive polypeptide drug construct according to any one of claims 1 to 12 , comprising: producing the bioactive polypeptide drug construct according to claim 18 under conditions that promote expression of the bioactive polypeptide drug construct. 1. A method comprising culturing host cells of and recovering a bioactive polypeptide drug construct protein.

An isolated bioactive polypeptide drug construct protein produced by the method of claim 19 .