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JP-2026076241-A - IL15/IL15Rα heterodimer FC-fusion protein

JP2026076241AJP 2026076241 AJP2026076241 AJP 2026076241AJP-2026076241-A

Abstract

[Problem] To provide a fusion protein that has been engineered to address the short half-life of the IL-15/IL-15Rα heterodimer, which can potently activate T cells. [Solution] A novel IL15/IL15Rα heterodimer Fc fusion protein having a specific amino acid sequence is provided. Methods for producing these proteins and methods for treating patients with these proteins are also provided, along with nucleic acids, expression vectors, and host cells. [Selection Diagram] Figure 1

Inventors

  • バーネット，マシュー
  • ラシッド，ルマナ
  • デスジャルレイス，ジョン
  • ヴァルマ，ラジャット
  • ボンゾン，クリスティーン

Assignees

  • ゼンコア インコーポレイテッド

Dates

Publication Date
20260511
Application Date
20260121
Priority Date
20161014

Claims (20)

  1. It is a heterodimer protein, a) A first fusion protein comprising a first protein domain and a first Fc domain, wherein the first protein domain is covalently attached to the N-terminus of the first Fc domain using a first domain linker, b) A second fusion protein comprising a second protein domain and a second Fc domain, wherein the second protein domain is covalently attached to the N-terminus of the Fc domain using a second domain linker, A heterodimer protein in which the first Fc domain and the second Fc domain have a set of amino acid substitutions selected from the group consisting of S267K/L368D/K370S:S267K/LS364K/E357Q, S364K/E357Q:L368D/K370S, L368D/K370S:S364K, L368E/K370S:S364K, T411T/E360E/Q362E:D401K, L368D/K370S:S364K/E357L, and K370S:S364K/E357Q, wherein the first protein domain contains IL15 protein and the second protein domain contains IL15Rα protein.
  2. The heterodimer protein according to claim 1, wherein the first Fc domain and/or the second Fc domain have an additional set of amino acid substitutions, including Q295E/N384D/Q418E/N421D, according to EU numbering.
  3. The heterodimer protein according to claim 1 or 2, wherein the first Fc domain and/or the second Fc domain have an additional set of amino acid substitutions consisting of G236R/L328R, E233P/L234V/L235A/G236del/S239K, E233P/L234V/L235A/G236del/S267K/A327G, E233P/L234V/L235A/G236del/S267K/A327G, and E233P/L234V/L235A/G236del.
  4. The heterodimer protein according to any one of claims 1 to 3, wherein the IL15 protein has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (full-length human IL15) and SEQ ID NO: 2 (shortened human IL15), and the IL15Rα protein has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (full-length human IL15Rα) and SEQ ID NO: 4 (sushi domain of human IL15Rα).
  5. The heterodimer protein according to any one of claims 1 to 4, wherein the IL15 protein has one or more amino acid substitutions selected from the group consisting of N1D, N4D, D8N, D30N, D61N, E64Q, N65D, and Q108E.
  6. The heterodimer protein according to any one of claims 1 to 5, wherein the IL15 protein and the IL15Rα protein each have a set of amino acid substitutions or additions selected from the group consisting of E87C:D96/P97/C98, E87C:D96/C97/A98, V49C:S40C, L52C:S40C, E89C:K34C, Q48C:G38C, E53C:L42C, C42S:A37C, and L45C:A37C.
  7. The aforementioned heterodimer protein, i) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP15902), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15908), ii) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP15902), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15909), iii) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP16479), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15908), iv) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP15902), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP16481), v) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP15902), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP16483), vi) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP16479), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15909), vii) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP16479), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP16481), viiii) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP16480), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP16482), ix) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP16480), and the second fusion protein h having the polypeptide sequence of Sequence ID XX (XENP15909), x) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP17064), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP17038), xi) The first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP17064), and the second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP17040), or xi) The first fusion protein having the polypeptide sequence of SEQ ID NO: XX (17062), and the second fusion protein having the polypeptide sequence of SEQ ID NO: XX (17044), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17686), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17687), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), xv) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP17688), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15908), xvi) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP17689), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15908), xvii) The first fusion protein having the polypeptide sequence of Sequence ID XX (XENP17690), and the second fusion protein having the polypeptide sequence of Sequence ID XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17691), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17692), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17693), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17694), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17695), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX(XENP17696), and the second fusion protein having the polypeptide sequence of sequence number XX(XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX(XENP17697) and the second fusion protein having the polypeptide sequence of sequence number XX(XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17698), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17699), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17701), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17691), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17702), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17703), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17704), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP17705), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number xxxiii)XX (XENP18295), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP17761), The first fusion protein having the polypeptide sequence of sequence number xxxiv) XX (XENP18783), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP18784), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP18786), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), The first fusion protein having the polypeptide sequence of sequence number XX (XENP18788), and the second fusion protein having the polypeptide sequence of sequence number XX (XENP15908), A heterodimer protein according to any one of claims 1 to 6, comprising: a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP19242) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP16481); or a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP19243) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP16481).
  8. The heterodimer protein according to any one of claims 1 to 7, wherein the first protein domain is directly and covalently attached to the N-terminus of the first Fc domain without using the first domain linker, and/or the second protein domain is directly and covalently attached to the N-terminus of the second Fc domain without using the second domain linker.
  9. The aforementioned heterodimer proteins are XENP20818, XENP20819, XENP21471, XENP21472, XENP21473, XENP21474, XENP21475, XENP21476, XENP21477, XENP22013, XENP22815, XE NP22816, XENP22817, XENP22818, XENP22819, XENP22820, XENP22821, XENP2 2822, XENP22823, XENP22824, XENP22825, XENP22826, XENP22827, XENP22828 A heterodimer protein according to any one of claims 1 to 6, selected from the group consisting of XENP22829, XENP22830, XENP22831, XENP22832, XENP22833, XENP22834, XENP23343, XENP23504, XENP23554, XENP23555, XENP23557, XENP23559, XENP24019, XENP24020, XENP24045, XENP24051, XENP24052, XENP24113, XENP24301, XENP24306, and XENP24341.
  10. A nucleic acid composition encoding the first fusion protein according to any one of claims 1 to 9.
  11. A nucleic acid composition encoding the second fusion protein according to any one of claims 1 to 10.
  12. An expression vector comprising the nucleic acid composition described in claim 10.
  13. An expression vector comprising the nucleic acid composition described in claim 11.
  14. The expression vector according to claim 13, further comprising the nucleic acid composition according to claim 10.
  15. A host cell comprising one or more expression vectors according to any one of claims 12 to 14.
  16. It is a heterodimer protein, a) A fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is covalently attached to the N-terminus of the second protein domain using a first domain linker, and the second protein domain is covalently attached to the N-terminus of the first Fc domain using a second domain linker, b) Including a second Fc domain, A heterodimer protein in which the first Fc domain and the second Fc domain have a set of amino acid substitutions selected from the group consisting of S267K/L368D/K370S:S267K/LS364K/E357Q, S364K/E357Q:L368D/K370S, L368D/K370S:S364K, L368E/K370S:S364K, T411T/E360E/Q362E:D401K, L368D/K370S:S364K/E357L, and K370S:S364K/E357Q, wherein the first protein domain contains IL15Rα protein and the second protein domain contains IL15 protein.
  17. The heterodimer protein according to claim 16, wherein the first Fc domain and/or the second Fc domain have an additional set of amino acid substitutions, including Q295E/N384D/Q418E/N421D, according to EU numbering.
  18. The heterodimer protein according to claim 16 or 17, wherein the first Fc domain and/or the second Fc domain have an additional set of amino acid substitutions consisting of G236R/L328R, E233P/L234V/L235A/G236del/S239K, E233P/L234V/L235A/G236del/S267K/A327G, E233P/L234V/L235A/G236del/S267K/A327G, and E233P/L234V/L235A/G236del.
  19. The heterodimer protein according to any one of claims 16 to 18, wherein the IL15 protein has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (full-length human IL15) and SEQ ID NO: 2 (shortened human IL15), and the IL15Rα protein has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (full-length human IL15Rα) and SEQ ID NO: 4 (sushi domain of human IL15Rα).
  20. The heterodimer protein according to any one of claims 16 to 19, wherein the IL15 protein and the IL15Rα protein each have a set of amino acid substitutions selected from the group consisting of E87C:D96/P97/C98, E87C:D96/C97/A98, V49C:S40C, L52C:S40C, E89C:K34C, Q48C:G38C, E53C:L42C, C42S:A37C, and L45C:A37C.

Description

Priority Claim This application claims priority to U.S. Patent Application No. 62/408,655 filed on 14 October 2016, No. 62/416,087 filed on 1 November 2016, No. 62/443,465 filed on 6 January 2017, and No. 62/477,926 filed on 28 March 2017, which are expressly incorporated herein by reference in their entirety, with particular reference to the drawings, descriptions, and claims contained herein. IL-2 and IL-15 function to promote the proliferation and differentiation of B cells, T cells, and NK cells. IL-2 is also essential for the function and survival of regulatory T cells (Tregs). Both cytokines exert their cellular signaling functions through binding to a trimer complex consisting of two shared receptors: a common gamma chain (γc, CD132) and an IL-2 receptor B chain (IL-2Rβ, CD122), as well as an alpha chain specific to each cytokine, namely IL-2 receptor alpha (IL-2Rα, CD25) or IL-15 receptor alpha (IL-15Rα, CD215). Both cytokines are considered potentially valuable therapeutic agents in oncology, and IL-2 is approved for use in patients with metastatic renal cell carcinoma and malignant melanoma. Currently, there is no approved use for recombinant IL-15, although several clinical trials are underway. IL-2 presents several challenges as a therapeutic agent. Firstly, IL-2 preferentially activates T cells expressing a high-affinity receptor complex dependent on CD25 expression. Since Treg cells constitutively express CD25, they compete with effector T cells for IL-2 supply, and their activation is desirable for oncological treatment. This imbalance leads to the concept of high-dose IL-2. However, this approach presents further problems due to IL-2-mediated toxicity, such as vasoleakage syndrome. IL-2 is primarily secreted by activated T cells, and its receptor is present on activated T cells, Treg cells, NK cells, and B cells. In contrast, IL-15 is produced by monocytes and dendritic cells and is presented primarily as a membrane-bound heterodimer complex with IL-15Rα, which is also present on the same cells. Its effect is achieved by trans-presenting the IL-15/IL-15Rα complex to NK cells and CD8+ T cells expressing IL-2Rβ and a common gamma chain. As promising drugs, both cytokines suffer from extremely rapid clearance, with half-lives measured in minutes. Furthermore, IL-15 itself has low stability due to its preference for the IL-15Rα-associated complex. Recombinantly produced IL-15/IL-15Rα heterodimers have also been shown to potently activate T cells. Nevertheless, their short half-lives hinder desirable drug delivery. This invention solves this problem by providing a novel IL-15/IL-15Rα heterodimer Fc fusion protein. Accordingly, in one embodiment, the present invention provides a heterodimer protein comprising: a) a first fusion protein comprising a first protein domain and a first Fc domain, wherein the first protein domain is covalently bonded to the N-terminus of the first Fc domain using a first domain linker; and b) a second fusion protein comprising a second protein domain and a second Fc domain, wherein the second protein domain is covalently attached to the N-terminus of the Fc domain using a second domain linker. The domain has a set of amino acid substitutions selected from the group consisting of S267K/L368D/K370S:S267K/LS364K/E357Q, S364K/E357Q:L368D/K370S, L368D/K370S:S364K, L368E/K370S:S364K, T411T/E360E/Q362E:D401K, L368D/K370S:S364K/E357L, and K370S:S364K/E357Q, wherein the first protein domain contains the IL15 protein and the second protein domain contains the IL15Rα protein. In some embodiments, the first protein domain is directly covalently bonded to the N-terminus of the first Fc domain without the use of a first domain linker, and/or the second protein domain is directly covalently bonded to the N-terminus of the second Fc domain without the use of a second domain linker. In some embodiments, the heterodimer protein is (i) a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15902) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP5908), (ii) a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15902) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15909), (iii) a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP16479) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15908), (iv) a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15902) and a polypeptide sequence of SEQ ID NO: XX (XENP16481) (v) A second fusion protein having a sequence, a first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP15902) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP16483), (vi) A first fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP16479) and a second fusion protein having the polypeptide sequence of SEQ ID NO: XX (XENP1590