CN-115103686-B - IL-2 orthologs and methods of use thereof

Abstract

The present invention relates to hIL2 orthologs ("IL 2 orthologs") that specifically and selectively bind to extracellular domain (ECD) transmembrane polypeptides comprised of modified hCD122 polypeptides. Binding of the hIL2 ortholog to the modified hCD122 polypeptide is involved in the transduction pathway of intracellular signaling, resulting in the biological activity of the native intracellular signaling pattern associated with binding of hIL2 to the medium-or high-affinity hIL2 receptor, but exhibits selectivity for the engineered expressed hCD122 orthogonal receptor. The hIL2 ortholog of the invention shows significantly reduced binding compared to its binding to the extracellular domain of wild-type hCD122, alone, or when hCD122 is present in the form of an endogenous high or medium affinity hIL2 receptor.

Inventors

• P-J. Penaflor Aspria
• S.A. McCorey
• M. OTT
• S cowder

Assignees

• 辛德凯因股份有限公司

Dates

Publication Date

20260505

Application Date

20201211

Priority Date

20191213

Claims (5)

1. 1. An hIL2 ortholog which is a polypeptide having the amino acid sequence shown below: PTSSSTKKTQLQLSQLLVLLKAILNGINNYKNPKLTRM LTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSK NFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIV EFLNRWITFCQSIISTLT。
2. 2. An ex vivo method of making an engineered T cell product comprising at least 20% T cells expressing an orthogonal hCD122 receptor (hoCD 122), the method comprising the steps of: a. Isolating a population of T cells from a mammalian subject; b. contacting the isolated population of T cells with a recombinant vector comprising a nucleic acid sequence encoding hoCD122,122 operably linked to one or more expression control sequences such that expression of hoCD122 in mammalian T cells is promoted while allowing uptake of the recombinant vector by T cells; c. Contacting the isolated T cell population of step b with an effective amount of the hIL2 ortholog of claim 1.
3. 3. The method of claim 2, wherein prior to step (b), the population of T cells isolated from step (a), is contacted with an agent that stimulates a signal associated with a CD3 TCR complex, and an agent that stimulates a co-stimulatory molecule on the surface of the T cells.
4. 4. A cell population product made by the method of claim 2 or 3, wherein the cell population comprises at least 20% engineered hoCD T cells.
5. 5. A cell population product made by the method of claim 2 or 3, wherein the T cells are CAR-T cells or TILs.

Description

IL-2 orthologs and methods of use thereof Cross reference to related applications The present application claims priority from U.S. provisional patent application Ser. No. 62/948066 filed on 12/13 in 2019. Statement regarding government funds No funds of the united states government are used in the conception or in the practice of the subject matter of this invention. Background Controlled manipulation of differentiation, development and proliferation of cells, particularly engineered immune cells, is of great clinical significance. T cells have been engineered for therapeutic applications, such as the recognition and killing of cancer cells, intracellular pathogens, and cells involved in autoimmunity. The use of engineered cell therapies in the treatment of cancer is facilitated by selectively activating and expanding engineered T cells that provide specific functions and are directed to selectively attack cancer cells. In some examples of adoptive immunotherapy, T cells are isolated from the blood of a subject, treated ex vivo, and reinfused into the subject. Thus, there is a need for compositions and methods that can selectively activate targeted engineered cell populations. The challenge in manufacturing cell therapy products is that such "live drugs" require close control of their environment to maintain viability and function. In practice, isolated cells, whether from the patient (autologous) or from a single donor source (allogeneic), begin to rapidly lose function after leaving the subject or controlled culture conditions. Upon leaving the subject or controlled culture conditions, the health and function of the isolated cells is successfully maintained, allowing the isolated cells to resume function for reinsertion into the cell product manufacturing workflow or patient. Furthermore, a challenge in the clinical application of engineered T cell therapies is to selectively stimulate these engineered cells to maximize their therapeutic effect. A typical means of providing sustained activation of engineered T cell products is systemic administration of cytokines, such as IL2. However, systemic administration of IL2 is associated with non-specific stimulation outside of the engineered cell population, particularly at high doses, with significant toxicity in human subjects. In addition, IL2 has a short life in vivo and requires frequent administration of IL2 to maintain the engineered T cells in an activated state. Although the initially administered engineered cells from the initial population can be detected months or even years after administration of the engineered cell product, a significant portion of these engineered cells alone can be trapped in a resting state, requiring their reactivation to exhibit significant therapeutic effects. Thus, the challenge of cell-based therapies is to impart desirable adjustable behavior to the transferred cells, which is unaffected by endogenous signal transduction pathways, does not affect non-targeted endogenous cells, and can be selectively controlled after administration of the engineered cell population to a subject. CD122 is a component of the medium and high affinity IL2 receptor complex. Sockolosky, et al (Science (2018) 359:1037-1042) and Garcia, et al (U.S. patent application publication US2018/0228841A1, published on month 8, 16 of 2018) describe orthogonal IL2/CD122 ligand/receptor systems to facilitate selective stimulation of cells engineered to express orthogonal CD122 receptors. IL2 muteins that are cognate ligands for orthogonal receptors are also described. Engineered T cells expressing orthogonal (orthographic) CD122 are contacted with an orthogonal ligand ("IL 2 ortholog (IL 2 ortholog)") corresponding to such orthogonal CD122, such that they are capable of specifically activating such engineered T cells. In particular, such orthogonal IL2 receptor ligand complexes provide for the selective expansion of cells engineered to express orthogonal receptors in a mixed population of cells, particularly T cells. IL-2 orthologs with reduced affinity for the non-engineered medium affinity (CD 122/CD 132) IL-2 receptor complex or high affinity (CD 25/CD122/CD 132) IL-2 receptor complex are also used to selectively target the activity of ortholog IL-2 to cells exhibiting high expression of CD25, for example in the treatment of autoimmune diseases. IL-2 orthologs, which have significantly reduced affinity for the native wild-type hCD122 extracellular domain (ECD), but retain binding to the ECD of CD25, can also be used as competitive antagonists of wild-type IL-2 by interfering with the formation of high affinity IL-2 receptor complexes and thus can be used in the treatment of autoimmune diseases or Graft Versus Host (GVH) diseases. The present invention relates to ligands that interact with the orthogonal hCD122 receptor. In particular, there is provided an hIL-2 ortholog (hIL 2 ortholog) which provides selective binding and signal transduction through rece