DGKα/ζ inhibitors combine with PD-1 checkpoint therapy to promote T cell-mediated antitumor immunity.

Programmed cell death protein 1 (PD-1) immune checkpoint blockade therapy has revolutionized cancer treatment. Although PD-1 blockade is effective in a subset of patients with cancer, many fail to respond because of either primary or acquired resistance. Thus, next-generation strategies are needed to expand the depth and breadth of clinical responses. Toward this end, we designed a human primary T cell phenotypic high-throughput screening strategy to identify small molecules with distinct and complementary mechanisms of action to PD-1 checkpoint blockade. Through these efforts, we selected and optimized a chemical series that showed robust potentiation of T cell activation and combinatorial activity with αPD-1 blockade. Target identification was facilitated by chemical proteomic profiling with a lipid-based photoaffinity probe, which displayed enhanced binding to diacylglycerol kinase α (DGKα) in the presence of the active compound, a phenomenon that correlated with the translocation of DGKα to the plasma membrane. We further found that optimized leads within this chemical series were potent and selective inhibitors of both DGKα and DGKζ, lipid kinases that constitute an intracellular T cell checkpoint that blunts T cell signaling through diacylglycerol metabolism. We show that dual DGKα/ζ inhibition amplified suboptimal T cell receptor signaling mediated by low-affinity antigen presentation and low major histocompatibility complex class I expression on tumor cells, both hallmarks of resistance to PD-1 blockade. In addition, DGKα/ζ inhibitors combined with αPD-1 therapy to elicit robust tumor regression in syngeneic mouse tumor models. Together, these findings support targeting DGKα/ζ as a next-generation T cell immune checkpoint strategy.

The effect of cadmium on the growth and antioxidant response for freshwater algae Chlorella vulgaris.

The objective of the present work was to evaluate the effect of exogenously applied cadmium on the physiological response of green algae Chlorella vulgaris. The study investigated the long-term effect (18 days) of cadmium on the levels of algae biomass, assimilation pigment composition, soluble protein, oxidative status (production of hydrogen peroxide and superoxide anion), antioxidant enzymes (such as superoxide dismutase, peroxidase, catalase and glutathione reductase enzyme) in C. vulgaris. The results showed that growth, the amount of chlorophyll a (Chl a), chlorophyll b (Chl b) and carotenoids gradually decreased with increasing cadmium over 18 days exposure. Cadmium at concentration of 7 mg L(-1) inhibited algal growth expressed as the number of cells. Our research found that C. vulgaris has a high tolerance to cadmium. Contents of chlorophylls (Chl a and Chl b) and carotenoids (Car) of C. vulgaris was significantly decline with rising concentration of cadmium (p < 0.05). The decrease of 54.04 and 93.37 % in Chl a, 60.65 and 74.32 % in Chl b, 50.00 and 71.88 % in total carotenoids was noticed following the treatment with 3 and 7 mg L(-1) cadmium doses compared with control treatment, respectively. Cadmium treatments caused a significant change in the physiological competence (calculated as chlorophyll a/b) which increased with increasing Cd(II) doses up to 1 mg L(-1) but decreased at 3 mg L(-1). While accumulation of soluble protein was enhanced by presence of cadmium, the treatment with cadmium at 3 and 7 mg L(-1) increased the concentration of soluble proteins by 88, 95.8 % in C. vulgaris, respectively. Moreover, low doses of cadmium stimulated enzymatic (superoxide dismutase, catalase and glutathione reductase) in C. vulgaris, The content of peroxidase increased with the increasing cadmium concentration, and had slightly decreased at the concentration of 7 mg L(-1), but was still higher than control group, which showed that cadmium stress at high concentration mainly peroxidase works in C. vulgaris. And therefore, suppressed reactive oxygen species (hydrogen peroxide and superoxide) accumulated. The present study also showed that cadmium increased oxidative stress and induced antioxidant defense systems against reactive oxygen species. The observation in here analyzed C. vulgaris after exposure to cadmium indicate that hydrogen peroxide, superoxide and peroxidase in the alga with exposure to Cd(II) seemed to be parameters as biomarkers for metal-induced oxidative stress.

3,4-Diamino-2,5-thiadiazole-1-oxides as potent CXCR2/CXCR1 antagonists.

A series of novel and potent 3,4-diamino-2,5-thiadiazole-1-oxides were prepared and found to show excellent binding affinities for CXCR2 and CXCR1 receptors and excellent inhibitory activity of Gro-alpha and IL-8 mediated in vitro hPMN MPO release of CXCR2 and CXCR1 expressing cell lines. On the other hand, a closely related 3,4-diamino-2,5-thiadiazole-dioxide did not show functional activity despite its excellent binding affinities for CXCR2 and CXCR1 in membrane binding assays. A detailed SAR has been discussed in these two closely related structures.

Identification of full, partial and inverse CC chemokine receptor 3 agonists using [35S]GTPgammaS binding.

Study of the CC chemokine receptor 3 (CCR3) has been limited to using radiolabeled agonist chemokines. A small molecule CCR3 antagonist, 2-[(6-amino-2-benzothiazolyl)thio]-N-[1-[(3,4-dichlorylphenyl)methyl]-4-piperidinyl]acetamide, Banyu (I), was tritiated and used for pharmacological studies. Banyu (I) has a K(d) of 5.0+/-0.4 and 4.3+/-1.8 nM on human CCR3 transfectants and eosinophils, and noncompetitively inhibits [125I]eotaxin binding and eotaxin-induced [35S]guanosine-5'-O-(3-thiotriphosphate) ([35S]GTPgammaS) binding. The proportion of [125I]eotaxin: [3H]Banyu (I) binding sites in eosinophils or transfectants was 35% or 13%, although both binding sites were overexpressed in transfectants. CCR3 spontaneously couples to G-proteins in CCR3 transfectants, demonstrated by changes in basal and eotaxin-induced [35S]GTPgammaS binding under reduced NaCl and GDP concentrations. Consequently, Banyu (I) was identified as an inverse agonist. In contrast, CCL18 and I-TAC (interferon-inducible T cell alpha-chemoattractant) were neutral antagonists, inhibiting eotaxin-induced [35S]GTPgammaS binding, with minimal effect on basal coupling of CCR3 to G proteins. Eotaxin, eotaxin-2 and monocyte chemoattractant protein (MCP)-4 are full agonists inducing [35S]GTPgammaS binding; eotaxin-3, MCP-3, RANTES (regulated on activation normal T cell expressed and secreted), vMIP-I (Kaposi's sarcoma-associated herpesvirus macrophage inflammatory protein-) and vMIP-II are partial agonists, indicating that this is a sensitive method to quantitate agonist efficacy.