	US 12,029,778 B2
	Interleukin-18 mimics and methods of use
Aaron Ring, New Haven, CT (US)
Assigned to Yale University, New Haven, CT (US)
Filed by Yale University, New Haven, CT (US)
Filed on May 11, 2020, as Appl. No. 16/871,833.
Claims priority of provisional application 62/847,190, filed on May 13, 2019.
Prior Publication US 2021/0015891 A1, Jan. 21, 2021
Int. Cl. G01N 33/48 (2006.01); A61K 38/16 (2006.01); A61P 35/00 (2006.01); C07K 14/54 (2006.01); G06G 7/48 (2006.01); G06G 7/58 (2006.01)

CPC A61K 38/16 (2013.01) [A61P 35/00 (2018.01); C07K 14/54 (2013.01); G06G 7/48 (2013.01); G06G 7/58 (2013.01)]

32 Claims

1. A method of making a mimic of a parent IL-18 protein, the method comprising:

(a) computationally designing a polypeptide de novo from a parent IL-18 protein by:

(i) defining as a template a structure of the IL-18 parent protein in a complex with at least human IL-18Rα, wherein the parent IL-18 protein is a decoy resistant (DR) IL-18 variant polypeptide of wild-type (WT) human IL-18;

(ii) designating one or more hotspots in the template based on binding sites in the complex;

(iii) inputting the template comprising the one or more designated hotspots into a mimetic design protocol to generate a de novo polypeptide backbone; and

(iv) outputting an amino acid sequence for the de novo designed polypeptide; and

(b) producing the de novo designed polypeptide that was computationally designed in step (a), wherein the amino acid sequence of the produced de novo designed polypeptide has 84% or less sequence identity with the WT human IL-18 SEQ ID NO:30, has a beta trefoil fold, specifically binds to human IL-18 receptor (IL-18R), and exhibits substantially reduced binding to human IL-18 binding protein (IL-18BP) as compared with WT human IL-18 with a K_Dfor IL-18BP measured by SPR that is 10 nM or greater.