gw2974 and Disease-Models--Animal

When Zika virus emerged as a public health emergency there were no drugs or vaccines approved for its prevention or treatment. We used a high-throughput screen for Zika virus protease inhibitors to identify several inhibitors of Zika virus infection. We expressed the NS2B-NS3 Zika virus protease and conducted a biochemical screen for small-molecule inhibitors. A quantitative structure-activity relationship model was employed to virtually screen ∼138,000 compounds, which increased the identification of active compounds, while decreasing screening time and resources. Candidate inhibitors were validated in several viral infection assays. Small molecules with favorable clinical profiles, especially the five-lipoxygenase-activating protein inhibitor, MK-591, inhibited the Zika virus protease and infection in neural stem cells. Members of the tetracycline family of antibiotics were more potent inhibitors of Zika virus infection than the protease, suggesting they may have multiple mechanisms of action. The most potent tetracycline, methacycline, reduced the amount of Zika virus present in the brain and the severity of Zika virus-induced motor deficits in an immunocompetent mouse model. As Food and Drug Administration-approved drugs, the tetracyclines could be quickly translated to the clinic. The compounds identified through our screening paradigm have the potential to be used as prophylactics for patients traveling to endemic regions or for the treatment of the neurological complications of Zika virus infection.

Topics: Animals; Antiviral Agents; Artificial Intelligence; Chlorocebus aethiops; Disease Models, Animal; Drug Evaluation, Preclinical; High-Throughput Screening Assays; Immunocompetence; Inhibitory Concentration 50; Methacycline; Mice, Inbred C57BL; Protease Inhibitors; Quantitative Structure-Activity Relationship; Small Molecule Libraries; Vero Cells; Zika Virus; Zika Virus Infection

There is a major clinical need for new therapies for the treatment of chronic itch. Many of the molecular components involved in itch neurotransmission are known, including the neuropeptide NPPB, a transmitter required for normal itch responses to multiple pruritogens in mice. Here, we investigated the potential for a novel strategy for the treatment of itch that involves the inhibition of the NPPB receptor NPR1 (natriuretic peptide receptor 1). Because there are no available effective human NPR1 (hNPR1) antagonists, we performed a high-throughput cell-based screen and identified 15 small-molecule hNPR1 inhibitors. Using in vitro assays, we demonstrated that these compounds specifically inhibit hNPR1 and murine NPR1 (mNPR1). In vivo, NPR1 antagonism attenuated behavioral responses to both acute itch- and chronic itch-challenged mice. Together, our results suggest that inhibiting NPR1 might be an effective strategy for treating acute and chronic itch.

Topics: Animals; Behavior, Animal; Cell-Free System; Dermatitis, Contact; Disease Models, Animal; Ganglia, Spinal; Humans; Mice, Inbred C57BL; Mice, Knockout; Neurons; Pruritus; Receptors, Atrial Natriuretic Factor; Reproducibility of Results; Signal Transduction; Small Molecule Libraries

To evaluate the inhibiting effects of GW2974, a tyrosine kinase inhibitors, on dimethyl-benzanthracene (DMBA)-induced Syrian golden hamster buccal pouch carcinogenesis.. Ninety hamsters were painted with 0.5% DMBA in the left buccal pouches three times a week for 6 weeks, which were then divided into 3 groups: low-concentration, high-concentration and positive control groups. Positive control group received no further treatment. Ten hamsters served as negative control. The two treated groups were topically painted with GW2974 (4 mmol/L) and GW2974 (8 mmol/L) three times a week, respectively. Tissue samples of the left cheek pouch were obtained at 24 th week. The average number, average volume and burden of tumor, incidence of tumor and the pathological changes of each group were recorded.. After GW2974 (4 mmol/L and 8 mmol/L) was applied topically, tumor incidence decreased from 80.0% (24/30) to 43.3% (13/30, P < 0.01) and 36.7% (11/30, P < 0.01) respectively, the average number of tumors decreased from 1.00 +/- 0.87 to 0.47 +/- 0.82 (P < 0.05) and 0.37 +/- 0.62 (P < 0.05), the average volume and burden of tumors also declined, tumor incidence decreased from 70.0% (21/30) to 40.0% (12/30, P < 0.05) and 33.3% (10/30, P < 0.01), the number of tumors declined from 1.83 +/- 1.91 to 0.67 +/- 0.99 (P < 0.05) and 0.43 +/- 0.68 (P < 0.01).. Topical application of GW2974 could significantly inhibit hamster buccal pouch carcinogenesis, which suggests that GW2974 may have a major impact in chemoprevention and treatment of oral cancer.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cricetinae; Disease Models, Animal; Male; Mouth Neoplasms; Protein-Tyrosine Kinases; Quinazolines

Biliary tract cancer (BTC) is the second most common primary hepatobiliary cancer after hepatocellular cancer. At the time of diagnosis, most BTC are at an advanced stage and are unresectable. There is presently no effective curative treatment of the advanced disease nor is there any effective clinical therapy that will prevent the development of BTC. All of these factors render gallbladder cancer nearly incurable with a poor survival rate. The aim of our study was to provide a better understanding of the mechanisms involved in the development of gallbladder carcinoma as the advancement of more effective treatment options would significantly improve prognosis. In the present study, we examined the effect of gefitinib, a selective epidermal growth factor receptor/tyrosine kinase inhibitor (EGFR/TKI), on the development of gallbladder carcinoma in BK5.erbB2 mice. In addition, we examined the effect of another quinazoline derivative, GW2974, which is able to block the activation of both the EGFR and erbB2, in this model. Animals were treated with either 400 ppm gefitinib or 200 ppm GW2974 as a supplement in the diet using either a chemopreventive or therapeutic protocol. The results show that both compounds were potent chemopreventive and therapeutic agents in this mouse model of human BTC. The results also suggest that activation of the EGFR plays an important role in development of BTC in this model and that targeting both the EGFR and erbB2 may be an effective strategy for treatment of this disease.

Topics: Animals; Anticarcinogenic Agents; Antineoplastic Agents; Blotting, Western; Body Weight; Carcinoma; Cell Line, Tumor; Disease Models, Animal; Enzyme Inhibitors; ErbB Receptors; Gallbladder; Gallbladder Neoplasms; Gefitinib; Humans; In Situ Nick-End Labeling; MAP Kinase Signaling System; Mice; Mice, Transgenic; Microscopy, Fluorescence; Neoplasms, Experimental; Phosphorylation; Protein-Tyrosine Kinases; Quinazolines; Receptor, ErbB-2; Tolonium Chloride

Described herein is the design and synthesis of indazolylaminopyridopyrimidines and quinazolines as inhibitors of the class 1 tyrosine kinase receptor family. Data is presented for N(4)-(1-benzyl-1H-indazol-5-yl)-N(6),N(6)-dimethylpyrido[3,4-d]pyrimidine-4,6-diamine 3B. This compound inhibited EGFr and c-erbB-2 enzymes selectively over other kinases. It inhibited the proliferation of a range of tumour cell lines in vitro and the growth of BT474 xenografts in SCID mice.

Topics: Animals; Antineoplastic Agents; Cell Division; Disease Models, Animal; ErbB Receptors; Mice; Mice, SCID; Pyrimidines; Quinazolines; Receptor, ErbB-2; Tumor Cells, Cultured; Xenograft Model Antitumor Assays