US 12,303,567 B2
Compositions of protein complexes and methods of use thereof
John Thomas Mulligan, Seattle, WA (US); Shannon Lee Okada, Seattle, WA (US); Justin Richard Killebrew, Seattle, WA (US); and Diane Louise Hollenbaugh, Seattle, WA (US)
Assigned to BONUM THERAPEUTICS, INC., Seattle, WA (US)
Filed by Bonum Therapeutics, Inc., Seattle, WA (US)
Filed on Feb. 10, 2023, as Appl. No. 18/108,514.
Application 18/108,514 is a continuation of application No. 17/504,385, filed on Oct. 18, 2021, granted, now 11,642,417.
Application 17/504,385 is a continuation of application No. PCT/US2021/032313, filed on May 13, 2021.
Claims priority of provisional application 63/024,422, filed on May 13, 2020.
Prior Publication US 2024/0024501 A1, Jan. 25, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. A61K 47/68 (2017.01); C07K 16/24 (2006.01); C07K 16/28 (2006.01)
CPC A61K 47/6813 (2017.08) [A61K 47/6845 (2017.08); A61K 47/6849 (2017.08); C07K 16/246 (2013.01); C07K 16/249 (2013.01); C07K 16/2818 (2013.01); C07K 2317/31 (2013.01)] 19 Claims
 
1. A method of selectively activating cytokine receptor signaling in a presence of a marker, the method comprising:
(a) obtaining a self-regulating complex that adopts an inactive state in an absence of the marker and an active state in the presence of the marker, the self-regulating complex comprising a cytokine and a sensor domain, wherein the cytokine is linked to the sensor domain by a flexible linker, wherein the sensor domain is a dual-binding antibody comprising a heavy chain variable domain and a light chain variable domain; wherein the sensor domain comprises a binding affinity for the cytokine and a binding affinity for the marker, wherein the cytokine and the marker compete for binding to the sensor domain and the sensor domain binds the cytokine and the marker, but not both simultaneously;
(b) binding the sensor domain to the cytokine, thereby blocking the cytokine from binding to its cytokine receptor and rendering the cytokine inactive;
(c) contacting the self-regulating complex to the marker, wherein upon the contacting:
(i) the sensor domain binds to the marker;
(ii) the sensor domain is blocked from binding the cytokine; and
(iii) the cytokine is capable of binding to its cytokine receptor; and
(d) contacting the self-regulating complex to the cytokine receptor, wherein the cytokine binds to the cytokine receptor, thereby selectively activating the cytokine receptor signaling in the presence of the marker;
wherein:
(I) the heavy chain variable domain and the light chain variable domain are present in a Fab of an immunoglobulin G (IgG) isotype antibody, wherein a C-terminus of the cytokine is joined to an N-terminus of the heavy chain variable domain or the light chain variable domain by the flexible linker;
(II) the heavy chain variable domain and the light chain variable domain are present in a single chain variable fragment (scFv), wherein the C-terminus of the cytokine is joined to the N-terminus of the heavy chain variable domain or the light chain variable domain by the flexible linker; or
(III) the heavy chain variable domain and the light chain variable domain are present in a scFv, wherein an N-terminus of the scFv is joined to a C-terminus of a first heavy chain of an IgG isotype antibody, and an N-terminus of the cytokine is joined to a C-terminus of a second heavy chain of the IgG isotype antibody;
wherein:
the marker is human Leucine Rich Repeat Containing 15 (LRRC15) and the cytokine is human interferon (IFN)-α;
the marker is human Carcino Embryonic Antigen (CEA) and the cytokine is human IFN-α;
the marker is human PD-L1 and the cytokine is human IFN-α;
the marker is human PD-1 and the cytokine is human interleukin (IL)-2; or
the marker is human PD-L1 and the cytokine is human IL-15.