After chromatographic purification and subsequent deprotection, we achieved a 39% yield of compound 9i

After chromatographic purification and subsequent deprotection, we achieved a 39% yield of compound 9i. the PD-1/PD-L1 axis has shown impressive clinical benefit, with durable regression and even cure in a subset of hard-to-treat cancers.2,3 In general, ICB-responsive cancers are characterized by high levels of mutations and corresponding neoantigens (hot tumors). These neoantigens can be recognized by immune effector T-cells, which under homeostatic conditions would result in cancer cell elimination.4 In cancer, this elimination is restrained by the immune checkpoint PD-1 expressed on T-cells. PD-1 binds to PD-L1 expressed on cancer cells, resulting in an inhibitory intracellular signaling cascade that prevents proper T-cell activation.5 Consequently, inhibition of the interaction of PD-L1 and PD-1 receptors promotes T-cell activation. Several PD-1/PD-L1-directed antibodies are in clinical use, and numerous experimental ones are under development. However, current PD-1/PD-L1-directed therapies are useful only for a small subset of patients, are expensive to produce, have a risk of adverse effects, and show development of resistance, which limits their utility.6,7 Therefore, novel therapeutic modalities such as small molecules or peptides exhibit a lot of promise.8,9 The only small-molecule inhibitor targeted against PD-L1 currently undergoing clinical trial is CA-170 (Figure ?Figure11B).10 However, by means of various functional cell assays it was recently proved not to be a direct binder to PD-1 or PD-L1, and its mode of action remains unclear.11 As part of our ongoing efforts to understand and develop small molecules that antagonize PD-1/PD-L1, we present here the design, synthesis, biological activity, and structural basis of imidazopyridines as PD-1/PD-L1 antagonists.8,11?14 Open in Mouse monoclonal to beta Actin.beta Actin is one of six different actin isoforms that have been identified. The actin molecules found in cells of various species and tissues tend to be very similar in their immunological and physical properties. Therefore, Antibodies againstbeta Actin are useful as loading controls for Western Blotting. However it should be noted that levels ofbeta Actin may not be stable in certain cells. For example, expression ofbeta Actin in adipose tissue is very low and therefore it should not be used as loading control for these tissues a separate window Figure 1 Design of PD-1/PD-L1 antagonists. (A) Generalized pharmacophore model of a PD-L1 antagonist. Aromatic (purple), hydrophobic (green), and basic (positive charged, blue) pharmacophores are included. (B) Examples of potent PD-L1 dimerizers taken from the patent literature. The pharmacophore is indicated by red and blue colors. (C) Design of imidazopyridines accessible by GBB-3CR. The four variable parts of the scaffold are indicated by the different colored boxes. (D) Modeling of an imidazopyridine into a PD-L1 dimer structure (PDB ID:5NIX). Our recently published cocrystal structures of several small molecules binding to PD-L1 have been used to propose a generalized pharmacophore model for small-molecule PD-L1 binders (Figure ?Figure11A).9,14?17 These structures triggered a wave of small-molecule designs and subsequent patent applications.18,19 Accordingly, a twisted biphenyl moiety is linked via a two-atom linker to a planar (hetero)aromatic ring fragment that has a methanamine para to the linker moiety. Symmetrical central biphenyl moieties with two times the linker (hetero)aromatic fragment have also been described as highly potent PD-L1 binders.20 The diversity of PD-L1 small-molecule scaffolds based on our proposed pharmacophore model and claimed in patents is great (Figure ?Figure11B).18,19 The biphenyl component allows for fewer variations, but the linker and (hetero)aromatic moieties can be executed in a variety of designs. Finally, the water-exposed part of the molecule allows for many variations useful to tune drug-like properties such as water solubility. To circumvent the lengthy and linear OP-3633 sequential syntheses of many small-molecule PD-1/PD-L1 antagonists, we decided to explore multicomponent reactions for the one-pot assembly of the central (hetero)aromatic part of the pharmacophore model. For this, we chose the GroebkeCBlackburnCBienaym reaction (GBB-3CR) which is a versatile three-component reaction of heterocyclic amidines, aldehydes, and isocyanides that gives access to drug-like molecules (Figure ?Figure11C).21?23 A key fragment of the scaffold is a bicyclic imidazo ring. OP-3633 In our design, the bicyclic heteroaromatic moiety (Figure ?Figure11 C, gray) is the central element of the scaffold, to which three suitable substituents are attached, namely, the biphenyl (Figure ?Figure11 C, blue), methanamine (Figure ?Figure11 C, yellow), and amino (Figure ?Figure11 C, OP-3633 green) moieties. To decide on the position of the substituents, we used molecular modeling performed via Moloc and Scorpion software (Figure ?Figure11D).24,25 The best fit into the receptor was to introduce the methanamine moiety by the GBB-3CR aldehyde component and the biphenyl moiety through a CCO coupling to the aminopyridine linker. Following the design and docking studies (Figure ?Figure11D), we opted for the imidazopyridine-containing scaffold (Figure ?Figure11C) and started on the development of a corresponding synthesis route that would yield the established scaffold. We envisioned a synthetic route in which the step with the highest introduction of variation should be carried out last. Therefore, the GBB reaction was chosen as one of the last stages. Consequently, the preparation of the biphenyl-substituted amidines was approached first (Scheme 1). We started synthesizing the twisted biphenyl moiety via the Suzuki cross-coupling reaction.