| 1. |
- Çevik Aras, Hülya, 1975, et al.
(författare)
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A non-peptide receptor inhibitor with selectivity for one of the neutrophil formyl peptide receptors, FPR 1.
- 2012
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Ingår i: Biochemical pharmacology. - : Elsevier BV. - 1873-2968 .- 0006-2952. ; 83:12, s. 1655-1662
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Tidskriftsartikel (refereegranskat)abstract
- The neutrophil formyl peptide receptors (FPR1 and FPR2) are members of the G-protein coupled receptor family. The signals generated by occupied FPRs are both pro-inflammatory and anti-inflammatory. Accordingly, these receptors have become a therapeutic target for the development of novel drugs that may be used to reduce injuries in inflammatory diseases including asthma, rheumatoid arthritis, Alzheimer's disease and cardiovascular diseases. To support the basis for a future pharmacological characterization, we have identified a small molecular non-peptide inhibitor with selectivity for FPR1. We used the FPR1 and FPR2 specific ligands fMLF and WKYMVM, respectively, and an earlier described ratio technique, to determine inhibitory activity combined with selectivity. We show that the compound 3,5-dichloro-N-(2-chloro-5-methyl-phenyl)-2-hydroxy-benzamide (BVT173187) fulfills the criteria for an FPR1 inhibitor selective for FPR1 over FPR2, and it inhibits the same functional repertoire in neutrophils as earlier described peptide antagonists. Accordingly, the new inhibitor reduced neutrophil activation with FPR1 agonists, leading to mobilization of adhesion molecules (CR3) and the generation of superoxide anion from the neutrophil NADPH-oxidase. The effects of a number of structural analogs were determined but these were either without activity or less active/specific than BVT173187. The potency of the new inhibitor for reduction of FPR1 activity was the same as that of the earlier described FPR1 antagonist cyclosporine H, but signaling through the C5aR and CXCR (recognizing IL8) was also affected by BVT173187.
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| 2. |
- Forsman, Huamei, et al.
(författare)
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Stable formyl peptide receptor agonists that activate the neutrophil NADPH-oxidase identified through screening of a compound library.
- 2011
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Ingår i: Biochemical pharmacology. - : Elsevier BV. - 1873-2968 .- 0006-2952. ; 81:3, s. 402-411
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Tidskriftsartikel (refereegranskat)abstract
- The neutrophil formyl peptide receptors (FPR1 and FPR2) are G-protein coupled receptors that can induce pro-inflammatory as well as anti-inflammatory activities when activated. Accordingly, these receptors may become therapeutic targets for the development of novel drugs to be used for reducing the inflammation induced injuries in asthma, rheumatoid arthritis, Alzheimer's disease, cardiovascular diseases and traumatic shock. We screened a library of more then 50K small compounds for an ability of the compounds to induce a transient rise in intracellular Ca(2+) in cells transfected to express FPR2 (earlier called FPRL1 or the lipoxin A(4) receptor). Ten agonist hits were selected for further analysis representing different chemical series and five new together with five earlier described molecules were further profiled. Compounds 1-10 gave rise to a calcium response in the FPR2 transfectants with EC(50) values ranging from 4×10(-9)M to 2×10(-7)M. All 10 compounds activated human neutrophils to release superoxide, and based on the potency of their activity, the three most potent activators of the neutrophil NADPH-oxidase were further characterized. These three agonists were largely resistant to inactivation by neutrophil produced reactive oxygen species and shown to trigger the same functional repertoire in neutrophils as earlier described peptide agonists. Accordingly they induced chemotaxis, granule mobilization and secretion of superoxide. Interestingly, the oxidase activity was largely inhibited by cyclosporine H, an FPR1 selective antagonist, but not by PBP10, an FPR2 selective inhibitor, suggesting that FPR1 is the preferred receptor in neutrophils for all three agonists.
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| 3. |
- Gad, Helge, et al.
(författare)
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MTH1 inhibition eradicates cancer by preventing sanitation of the dNTP pool
- 2014
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Ingår i: Nature. - : Nature Publishing Group. - 0028-0836 .- 1476-4687. ; 508:7495, s. 215-221
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Tidskriftsartikel (refereegranskat)abstract
- Cancers have dysfunctional redox regulation resulting in reactive oxygen species production, damaging both DNA and free dNTPs. The MTH1 protein sanitizes oxidized dNTP pools to prevent incorporation of damaged bases during DNA replication. Although MTH1 is non-essential in normal cells, we show that cancer cells require MTH1 activity to avoid incorporation of oxidized dNTPs, resulting in DNA damage and cell death. We validate MTH1 as an anticancer target in vivo and describe small molecules TH287 and TH588 as first-in-class nudix hydrolase family inhibitors that potently and selectively engage and inhibit the MTH1 protein in cells. Protein co-crystal structures demonstrate that the inhibitors bindin the active site of MTH1. The inhibitors cause incorporation of oxidized dNTPs in cancer cells, leading to DNA damage, cytotoxicity and therapeutic responses in patient-derived mouse xenografts. This study exemplifies the non-oncogene addiction concept for anticancer treatment and validates MTH1 as being cancer phenotypic lethal.
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| 4. |
- Kalderén, Christina, et al.
(författare)
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CCL2 mediates anti-fibrotic effects in human fibroblasts independently of CCR2
- 2014
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Ingår i: International Immunopharmacology. - : Elsevier BV. - 1567-5769 .- 1878-1705. ; 20:1, s. 66-73
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Tidskriftsartikel (refereegranskat)abstract
- CCL2 is known for its major role as a chemoattractant of monocytes for immunological surveillance and to site of inflammation. CCL2 acts mainly through the G-protein-coupled receptor CCR2 but has also been described to mediate its effects independently of this receptor in vitro and in vivo. Emerging pieces of evidence indicate that the CCL2/CCR2 axis is involved in fibrotic diseases, such as increased plasma levels of CCL2 and the presence of CCL2-hyperresponsive fibroblasts explanted from patients with systemic sclerosis and idiopathic pulmonary fibrosis. One of the profibrotic key mediators is the myofibroblast characterized by overexpression of alpha-smooth muscle actin and collagen I. However, the correlation between the CCL2/CCR2 axis and the activation of fibroblasts is not yet fully understood. We have screened human fibroblasts of various origins, human pulmonary fibroblasts (HPF), human fetal lung fibroblasts (HFL-1) and primary preadipocytes (SPF-1) in regard to CCL2 stimulated fibrotic responses. Surprisingly we found that CCL2 mediates anti-fibrotic effects independently of CCR2 in human fibroblasts of different origins.
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| 5. |
- Kalderén, Christina
(författare)
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Chemoattractant receptors in drug discovery : FPR2 and CCR2-two potential targets
- 2012
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Chemoattractants and chemoattractant receptors have many important functions in multicellular organisms, not least for their role in regulating migration of leukocytes. The receptors are also involved in many pathologies and they have, since they were cloned in the 1990ies, been regarded as attractive targets for development of drugs against cancer, viral infections, and inflammatory/autoimmune diseases. Although chemotactic molecules are very heterogeneous in terms of chemical structure most of them mediate their actions through binding to a G-protein-coupled receptor (GPCR). The formyl peptide receptors (FPRs) are chemoattractant receptors expressed on phagocytes and have important implications in host defense and inflammation as well as in resolution of inflammation. One of the goals was to identify FPR2 specific ligands with similar anti-inflammatory and proresolution properties as the eicosanoid lipoxin A4 previously described. A small compound library was screened for both agonist and antagonists and selected compounds were analyzed with respect to anti-inflammatory properties such as inhibition of NADPH oxidase activity and degranulation in primary neutrophils. Receptor selectivity was investigated by compound stimulation and desensitization studies in presence of receptor-specific tools. While the agonist screen resulted in non selective hit compounds with proinflammatory properties, the antagonist screen led to identification of anti-inflammatory agents with different selectivity. Another chemoattractant receptor binds CCL2, a member of the superfamily of chemokines. The function of CCL2 and its receptor (CCR2) is of importance for monocyte recruitment, and together this receptor ligand pair has many implications in inflammation. Accordingly, the importance of the CCL2/CCR2 axis in the recruitment of inflammatory cells and fibroblasts into fibrotic tissues was hypothesized as a mechanism for induction of fibrosis. There is, however, emerging evidence indicating that CCL2 may promote fibrosis also by other mechanisms such as activation of the fibroblast. The myofibroblast, synthesizing large amounts of extracellular matrix components (ECM) in response to profibrotic cytokines and growth factors, is the key cell in fibrosis. The effects of CCL2 on human fibroblasts with respect to ECM expression were studied and it was found that CCL2 might have a more multifaceted role in fibroblast activation than previously described. The truncated form of CCL2, CCL2 (1,9-76), also known as 7ND, is a receptor antagonist and is described to have therapeutic effects in several animal disease models such as bleomycin-induced lung fibrosis. The interaction between the 7ND and the human as well as the murine CCR2 was studied in vitro and data showed that 7ND was an inhibitor of human CCR2 but a very low affinity binder of the murine receptor. 7ND was, however, shown to downregulate fibrotic markers in murine fibroblasts, but in accordance with its inability to bind CCR2 the downregulation was found to be independent of this receptor.
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| 6. |
- Kitambi, Satish Srinivas, et al.
(författare)
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Vulnerability of Glioblastoma Cells to Catastrophic Vacuolization and Death Induced by a Small Molecule
- 2014
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Ingår i: Cell. - : Elsevier BV. - 0092-8674 .- 1097-4172. ; 157:2, s. 313-328
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Tidskriftsartikel (refereegranskat)abstract
- Glioblastoma multiforme (GBM) is the most aggressive form of brain cancer with marginal life expectancy. Based on the assumption that GBM cells gain functions not necessarily involved in the cancerous process, patient-derived glioblastoma cells (GCs) were screened to identify cellular processes amenable for development of targeted treatments. The quinine-derivative NSC13316 reliably and selectively compromised viability. Synthetic chemical expansion reveals delicate structure-activity relationship and analogs with increased potency, termed Vacquinols. Vacquinols stimulate death by membrane ruffling, cell rounding, massive macropinocytic vacuole accumulation, ATP depletion, and cytoplasmic membrane rupture of GCs. The MAP kinase MKK4, identified by a shRNA screen, represents a critical signaling node. Vacquinol-1 displays excellent in vivo pharmacokinetics and brain exposure, attenuates disease progression, and prolongs survival in a GBM animal model. These results identify a vulnerability to massive vacuolization that can be targeted by small molecules and point to the possible exploitation of this process in the design of anticancer therapies.
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