Granted Patent B2

US 12,624,087 · App. 18/046,859 · Granted May 12, 2026

Inventors: Ryan Swanson (Seattle, WA); Michael Kornacker (Seattle, WA); Daniel William Demonte (Seattle, WA); Mark F. Maurer (Seattle, WA)

Assignee: Alpine Immune Sciences, Inc.

C07K14/70596A61K35/17A61K40/10A61K40/11A61K40/24A61K40/42A61K40/421A61P35/00C07K14/70532C07K14/70535C07K16/28C12N15/62C12N15/85A61K40/50

Abstract

Claims (24)

1 . A variant CD155 polypeptide comprising an amino acid sequence that exhibits at least 90% sequence identity to SEQ ID NO: 47 or amino acid residues 4-199 of SEQ ID NO: 47, wherein the variant CD155 polypeptide comprises the amino acid substitution L104Q with reference to positions set forth in SEQ ID NO:47 of unmodified CD155;

wherein the variant CD155 polypeptide specifically binds to the ectodomain of TIGIT with increased affinity compared to that of the unmodified CD155.

2 . The variant CD155 polypeptide of claim 1 , further comprising one or more amino acid substitutions selected from G7E, D8G, V9A, V9D, V9I, V9L, V10F, V10G, V10I, V11A, V11E, V11M, Q12H, Q12K, Q12L, A13E, A13R, T15I, T15S, Q16H, P18C, P18F, P18H, P18L, P18S, P18T, P18Y, G19D, F201, F20S, F20Y, L21S, L21M, G22S, D23A, D23G, D23N, D23Y, S24A, S24P, V25A, V25E, T26M, C29R, Y30C, Y30F, Y30H, Q32L, Q32R, V33M, P34S, N35D, N35F, N35S, M36I, M36R, M36T, E37G, E37P, E37S, E37V, V38A, V38G, T39A, T39S, H40Q, H40R, H40T, V41A, V41M, S42A, S42C, S42G, S42L, S42N, S42P, S42Q, S42T, S42V, S42W, L44P, L44V, T45A, T45G, T45I, T45S, T45Q, T45V, W46C, W46R, A47E, A47G, A47V, R48Q, H49L, H49Q, H49R, G50S, E51G, E51K, E51V, S52A, S52E, S52G, S52K, S52L, S52M, S52P, S52Q, S52R, S52T, S52W, G53R, S54C, S54G, S54H, S54N, S54R, M55I, M55L, M55V, A56V, V57A, V57L, V57T, F58L, F58Y, H59E, H59N, N59R, Q60H, Q60K, Q60P, Q60R, T61A, T61G, T61K, T61M, T61R, T61S, Q62F, Q62H, Q62K, Q62L, Q62M, Q62R, Q62Y, P64S, S65A, S65C, S65G, S65D, S65T, S65Y, S65H, S65N, S65T, S65W, S67A, S67E, S67G, S67H, S67L, S67T, S67V, S67W, E68D, E68G, S69L, S69P, K70E, K70R, K70Q, L72Q, E73D, E73G, E73R, V75A, V75L, A76E, A76G, A76T, A77T, A77V, R78G, R78K, R78S, L79P, L79Q, L79V, G80D, G80S, A81E, A81P, A81T, A81V, E82D, E82G, L83P, L83Q, R84W, N85D, N85Y, N87T, L88P, R89K, M90I, M90L, M90V, F91S, F91T, F91P, G92A, G92E, G92W, R94H, V95A, E96D, D97G, E98D, E98S, G99D, G99Y, N100Y, T102S, V106A, V106I, V106L, T107A, T107L, T107M, T107S, T107V, F108H, F108L, F108Y, Q110R, G111D, G111R, S112I, S112N, S112V, R113G, R113W, S114N, S114T, V115A, V115M, D116G, or D116N, or a conservative amino acid substitution thereof.

3 . The variant CD155 polypeptide of claim 1 , comprising amino acid substitutions selected from S42A/L104Q/G111R, S42A/S52Q/L104Q/G111R, S52M/L104Q, S42L/S52L/Q62F/L104Q, S42N/L104Q/G111R, L104Q, S42W/S52R/Q62Y/L104Q, S52R/Q62F/L104Q/G111R, S42G/S52L/Q62F/L104Q, T45Q/S52K/Q62F/L104Q/G111R, T45Q/S52Q/Q62Y/L104Q/G111R, P18S/S65W/S67A/M90V/V95A/L104Q/G111R, or P18S/S65W/S67A/L104Q/G111R.

4 . The variant CD155 polypeptide of claim 1 , comprising the sequence of amino acids set forth in any of SEQ ID NOS: 183-184, 188-189, 199, 204, 207, 209, 222, 242-243, 274 and 1269 or a specific binding fragment thereof, or a sequence of amino acids that exhibits at least 95% sequence identity to any of SEQ ID NOS: 183-184, 188-189, 199, 204, 207, 209, 222, 242-243, 274 and 1269 or a specific binding fragment thereof and that contains the one or more of the amino acid modifications thereof, or

the sequence of amino acids set forth in any of SEQ ID NOS: 280, 281, 285-286, 296, 301, 304, 306, 319, 339-340, 371, 377-378, 382-383, 393, 398, 401, 403, 416, 436-437, 468, 1270, and 1271 or a specific binding fragment thereof, or a sequence of amino acids that exhibits at least 95% sequence identity to any of SEQ ID NOS: 280, 281, 285-286, 296, 301, 304, 306, 319, 339-340, 371, 377-378, 382-383, 393, 398, 401, 403, 416, 436-437, 468, 1270, and 1271 or a specific binding fragment thereof and that contains the one or more of the amino acid modifications thereof.

5 . The variant CD155 polypeptide of claim 1 , wherein the variant CD155 polypeptide specifically binds to the ectodomain of CD226 or CD96 with increased affinity compared to the binding of the unmodified CD155 to the same ectodomain of, CD226 or CD96.

6 . A fusion protein comprising the variant CD155 polypeptide of claim 1 , wherein the variant CD155 polypeptide is linked to a multimerization domain.

7 . The fusion protein of claim 6 , wherein the multimerization domain is an Fc domain or a variant Fc domain with reduced effector function.

8 . A homodimer comprising two copies of the fusion protein of claim 6 .

9 . The fusion protein of claim 6 , wherein the variant CD155 polypeptide comprises the amino acid substitutions P18S/S65W/S67A/L104Q/G111R.

10 . The fusion protein of claim 6 , wherein the variant CD155 polypeptide comprises the sequence of amino acids set forth in SEQ ID NO: 1271.

11 . The variant CD155 polypeptide of claim 1 wherein the variant CD155 polypeptide is a transmembrane immunomodulatory protein further comprising a transmembrane domain.

12 . An immunomodulatory protein, comprising the variant CD155 of claim 1 linked to a second polypeptide comprising an immunoglobulin superfamily (IgSF) domain.

13 . A conjugate, comprising a variant CD155 polypeptide of claim 1 linked to a targeting moiety that specifically binds to a molecule on the surface of a cell.

14 . A nucleic acid molecule, encoding the variant CD155 polypeptide of claim 1 .

15 . A method of producing a variant CD155 polypeptide, comprising introducing the nucleic acid molecule of claim 14 or a vector comprising the nucleic acid molecule of claim 14 into a host cell under conditions to express the polypeptide in the host cell.

16 . A method of engineering a cell expressing a variant CD155 polypeptide, comprising introducing a nucleic acid molecule encoding the variant CD155 polypeptide of claim 1 into a host cell under conditions in which the polypeptide is expressed in the host cell.

17 . An engineered cell, comprising the variant CD155 polypeptide of claim 1 or a nucleic acid molecule encoding a sequence of amino acids comprising the variant CD155 polypeptide of claim 1 .

18 . The engineered cell of claim 17 , wherein the cell further comprises a chimeric antigen receptor (CAR) or an engineered T-cell receptor (TCR).

19 . An infectious agent, comprising a variant CD155 polypeptide of claim 1 or a nucleic acid molecule encoding the variant CD155 polypeptide of claim 1 .

20 . A pharmaceutical composition, the variant CD155 polypeptide of claim 1 ; an immunomodulatory protein or conjugate the variant CD155 polypeptide of claim 1 ; an engineered cell or infectious agent the variant CD155 polypeptide of claim 1 ; or a nucleic acid molecule encoding the variant CD155 polypeptide of claim 1 .

21 . The variant CD155 polypeptide of claim 1 , comprising the amino acid substitutions P18S/S65W/S67A/L104Q/G111R.

22 . The variant CD155 polypeptide of claim 1 , comprising the sequence of amino acids set forth in SEQ ID NO: 1271.

Assignments (2)

Continuity (7)

References Cited (400)