Discovery of Macrocycle-Based HPK1 Inhibitors for T-Cell-Based Immunotherapy.

Hematopoietic progenitor kinase 1 (HPK1) is a negative regulator of T-cell activation, and targeting HPK1 is considered a promising strategy for improving responses to antitumor immune therapies. The biggest challenge of HPK1 inhibitor design is to achieve a higher selectivity to GLK, an HPK1 homology protein as a positive regulator of T-cell activation. Herein, we report the design of a series of macrocycle-based HPK1 inhibitors via a conformational constraint strategy. The identified candidate compound 5i exhibited HPK1 inhibition with an IC50 value of 0.8 nM and 101.3-fold selectivity against GLK. Compound 5i also displayed good oral bioavailability (F = 27-49%) in mice and beagles and favorable metabolic stability (T1/2 > 186.4 min) in human liver microsomes. More importantly, compound 5i demonstrated a clear synergistic effect with anti-PD-1 in both MC38 (MSI) and CT26 (MSS) syngeneic tumor mouse models. These results showed that compound 5i has a great potential in immunotherapy.

[1]  Kuojun Zhang,et al.  The development of small-molecule inhibitors targeting HPK1. , 2022, European journal of medicinal chemistry.

[2]  Wei Huang,et al.  Discovery of 3-pyrazolyl-substituted pyrazolo[1,5-a]pyrimidine derivatives as potent TRK inhibitors to overcome clinically acquired resistance. , 2022, European journal of medicinal chemistry.

[3]  Q. You,et al.  Hematopoietic Progenitor Kinase 1 in Tumor Immunology: A Medicinal Chemistry Perspective. , 2022, Journal of medicinal chemistry.

[4]  Wei Huang,et al.  Conformational adjustment overcomes multiple drug-resistance mutants of tropomyosin receptor kinase. , 2022, European journal of medicinal chemistry.

[5]  Hoai An Le Thi,et al.  Discovery of Spiro-azaindoline Inhibitors of Hematopoietic Progenitor Kinase 1 (HPK1). , 2021, ACS medicinal chemistry letters.

[6]  Wei Huang,et al.  Discovery of Next-Generation Tropomyosin Receptor Kinase Inhibitors for Combating Multiple Resistance Associated with Protein Mutation. , 2021, Journal of medicinal chemistry.

[7]  Zhijun Sun,et al.  Improving antitumor immunity using antiangiogenic agents: Mechanistic insights, current progress, and clinical challenges , 2021, Cancer communications.

[8]  Ming Jiang,et al.  Developing a Novel Anticancer Gold(III) Agent to Integrate Chemotherapy and Immunotherapy. , 2021, Journal of medicinal chemistry.

[9]  I. Linney,et al.  Inhibitors of immuno-oncology target HPK1 – a patent review (2016 to 2020) , 2021, Expert opinion on therapeutic patents.

[10]  X. Fradera,et al.  Identification of Potent Reverse Indazole Inhibitors for HPK1. , 2021, ACS medicinal chemistry letters.

[11]  Alan S. Futran,et al.  Discovery of Orally Active Isofuranones as Potent, Selective Inhibitors of Hematopoetic Progenitor Kinase 1 , 2021, ACS medicinal chemistry letters.

[12]  E. Ileana-Dumbrava Unraveling the mysteries of microsatellite instability , 2021, Science Translational Medicine.

[13]  Haiyan Xu,et al.  Pharmacological inhibition of hematopoietic progenitor kinase 1 positively regulates T-cell function , 2020, PloS one.

[14]  Yonghui Wang,et al.  Adenosine A2A Receptor Antagonists for Cancer Immunotherapy. , 2020, Journal of medicinal chemistry.

[15]  D. Spring,et al.  Strategies for the Diversity-Oriented Synthesis of Macrocycles. , 2019, Chemical reviews.

[16]  Vinay Prasad,et al.  Estimation of the Percentage of US Patients With Cancer Who Are Eligible for and Respond to Checkpoint Inhibitor Immunotherapy Drugs , 2019, JAMA network open.

[17]  M. Cummings,et al.  Structure-Based Macrocycle Design in Small-Molecule Drug Discovery and Simple Metrics To Identify Opportunities for Macrocyclization of Small-Molecule Ligands. , 2019, Journal of medicinal chemistry.

[18]  G. Dranoff,et al.  Cancer Immunotherapy: Beyond Checkpoint Blockade , 2019, Annual Review of Cancer Biology.

[19]  BinQing Wei,et al.  Hematopoietic Progenitor Kinase-1 Structure in a Domain-Swapped Dimer. , 2019, Structure.

[20]  I. Mellman,et al.  The Kinase Activity of Hematopoietic Progenitor Kinase 1 Is Essential for the Regulation of T Cell Function. , 2018, Cell reports.

[21]  T. Tan,et al.  GLK-IKKβ signaling induces dimerization and translocation of the AhR-RORγt complex in IL-17A induction and autoimmune disease , 2018, Science Advances.

[22]  M. Ahluwalia,et al.  Management of Brain Metastases in the New Era of Checkpoint Inhibition , 2018, Current Neurology and Neuroscience Reports.

[23]  Jedd D. Wolchok,et al.  Cancer immunotherapy using checkpoint blockade , 2018, Science.

[24]  Pornpimol Charoentong,et al.  Targeting immune checkpoints potentiates immunoediting and changes the dynamics of tumor evolution , 2018, Nature Communications.

[25]  C. Lukacs,et al.  Germinal‐center kinase‐like kinase co‐crystal structure reveals a swapped activation loop and C‐terminal extension , 2017, Protein science : a publication of the Protein Society.

[26]  Wei Liu,et al.  Discovery of (10R)-7-amino-12-fluoro-2,10,16-trimethyl-15-oxo-10,15,16,17-tetrahydro-2H-8,4-(metheno)pyrazolo[4,3-h][2,5,11]-benzoxadiazacyclotetradecine-3-carbonitrile (PF-06463922), a macrocyclic inhibitor of anaplastic lymphoma kinase (ALK) and c-ros oncogene 1 (ROS1) with preclinical brain expos , 2014, Journal of medicinal chemistry.

[27]  T. Tan,et al.  The kinase GLK controls autoimmunity and NF-κB signaling by activating the kinase PKC-θ in T cells , 2011, Nature Immunology.

[28]  É. Marsault,et al.  Macrocycles are great cycles: applications, opportunities, and challenges of synthetic macrocycles in drug discovery. , 2011, Journal of medicinal chemistry.

[29]  H. Yee,et al.  Hematopoietic progenitor kinase 1 is a critical component of prostaglandin E2-mediated suppression of the anti-tumor immune response , 2009, Cancer Immunology, Immunotherapy.

[30]  S. Alzabin,et al.  Hematopoietic Progenitor Kinase 1 Is a Negative Regulator of Dendritic Cell Activation , 2009, The Journal of Immunology.

[31]  Stephen P. Hale,et al.  The exploration of macrocycles for drug discovery — an underexploited structural class , 2008, Nature Reviews Drug Discovery.

[32]  S. Sawasdikosol,et al.  Hematopoietic progenitor kinase 1 (HPK1) negatively regulates prostaglandin E2-induced fos gene transcription. , 2003, Blood.

[33]  OUP accepted manuscript , 2022, Briefings In Bioinformatics.

[34]  T. Tan,et al.  Hematopoietic progenitor kinase 1 negatively regulates T cell receptor signaling and T cell–mediated immune responses , 2007, Nature Immunology.