Ni–Pd Catalyzed Cyclization of Sulfanyl 1,6-Diynes: Synthesis of 1′‑Homonucleoside Analogues

The Ni–Pd catalyzed addition–cyclization of sulfanyl 1,6-diynes <b>2</b>–<b>9</b> with nucleobases is described. The reactions of <i>N</i>-tethered 1,6-diynes with <i>N</i>³-benzoylthymine, <i>N</i>⁴,<i>N</i>⁴-bis­(Boc)­cytosine, <i>N</i>³-benzoyluracil and <i>N</i>⁶,<i>N</i>⁶-bis­(Boc)­adenine exclusively afforded the pyrrolylmethyl and furylmethyl nucleotides in good yields. Deprotection of nucleobases was completed by treatment with acids or bases. Furthermore, the reactions of pyrroles and furans with nucleophiles such as alkoxides and amines underwent detosylation and conversion to the alkoxymethyl- and arylaminomethyl-pyrroles and furans in good yields

TNFAIP3 promotes survival of CD4 T cells by restricting MTOR and promoting autophagy

<p>Autophagy plays important roles in metabolism, differentiation, and survival in T cells. TNFAIP3/A20 is a ubiquitin-editing enzyme that is thought to be a negative regulator of autophagy in cell lines. However, the role of TNFAIP3 in autophagy remains unclear. To determine whether TNFAIP3 regulates autophagy in CD4 T cells, we first analyzed <i>Tnfaip3</i>-deficient naïve CD4 T cells in vitro. We demonstrated that <i>Tnfaip3</i>-deficient CD4 T cells exhibited reduced MAP1LC3/LC3 (microtubule-associated protein 1 light chain 3) puncta formation, increased mitochondrial content, and exaggerated reactive oxygen species (ROS) production. These results indicate that TNFAIP3 promotes autophagy after T cell receptor (TCR) stimulation in CD4 T cells. We then investigated the mechanism by which TNFAIP3 promotes autophagy signaling. We found that TNFAIP3 bound to the MTOR (mechanistic target of rapamycin) complex and that <i>Tnfaip3</i>-deficient cells displayed enhanced ubiquitination of the MTOR complex and MTOR activity. To confirm the effects of enhanced MTOR activity in <i>Tnfaip3</i>-deficient cells, we analyzed cell survival following treatment with Torin1, an MTOR inhibitor. <i>Tnfaip3</i>-deficient CD4 T cells exhibited fewer cell numbers than the control cells in vitro and in vivo. In addition, the impaired survival of <i>Tnfaip3</i>-deficient cells was ameliorated with Torin1 treatment in vitro and in vivo. The effect of Torin1 was abolished by <i>Atg5</i> deficiency. Thus, enhanced MTOR activity regulates the survival of <i>Tnfaip3</i>-deficient CD4 T cells. Taken together, our findings illustrate that TNFAIP3 restricts MTOR signaling and promotes autophagy, providing new insight into the manner in which MTOR and autophagy regulate survival in CD4 T cells.</p

Frequency scale for symptoms of gastroesophageal reflux disease.

<p>Questionnaire is consist of 12 questions including 7 (light blue lines) with acid reflux symptoms and 5 (dark blue lines) with dysmotility symptoms.</p

HR-QOL (SF-8 survey score) before and after smoking cessation.

<p>General health (GH), vitality (VT), and mental health (MH) significantly improved after smoking cessation in the success group, whereas there were no significant changes in the failure group. PF, physical functioning; RO, role physical; BP, bodily pain; SF, social functioning; RE, role emotional; PCS, physical component summary; MCS, mental component summary. *p<0.05 versus baseline.</p

Clinical characteristics of the study subjects at baseline.

<p>Clinical characteristics of the study subjects at baseline.</p

Factors associated with new development of gastroesophageal reflux disease.

<p>Factors associated with new development of gastroesophageal reflux disease.</p

Flow chart of study participants.

<p>Among 949 patients who initially visited smoking cessation clinics, 420 underwent smoking cessation therapy using varenicline for 12 weeks; all these patients ceased smoking in the short term. Twenty patients who used acid suppressive drugs, had peptic ulcer disease, or had a history of upper GI surgery were excluded, and 209 patients did not complete the survey 1 year later. Of 191 patients who completed the survey 1 year after attempted smoking cessation, 141 patients achieved smoking cessation (success group) and 50 did not (failure group).</p

Novel polyubiquitin imaging system, PolyUb-FC, reveals that K33-linked polyubiquitin is recruited by SQSTM1/p62

<p>Ubiquitin chains are formed with 8 structurally and functionally distinct polymers. However, the functions of each polyubiquitin remain poorly understood. We developed a polyubiquitin-mediated fluorescence complementation (PolyUb-FC) assay using Kusabira Green (KG) as a split fluorescent protein. The PolyUb-FC assay has the advantage that monoubiquitination is nonfluorescent and chain-specific polyubiquitination can be directly visualized in living cells without using antibodies. We applied the PolyUb-FC assay to examine K33-linked polyubiquitin. We demonstrated that SQSTM1/p62 puncta colocalized with K33-linked polyubiquitin and this interaction was modulated by the ZRANB1/TRABID-K29 and -K33 linkage-specific deubiquitinase (DUB). We further showed that the colocalization of K33-linked polyubiquitin and MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) puncta was impaired by SQSTM1/p62 deficiency. Taken together, these findings provide novel insights into how atypical polyubiquitin is recruited by SQSTM1/p62. Finally, we developed an inducible-PolyUb-FC system for visualizing chain-specific polyubiquitin. The PolyUb-FC will be a useful tool for analyzing the dynamics of atypical polyubiquitin chain generation.</p

Carbamoyl Pyridone HIV‑1 Integrase Inhibitors 3. A Diastereomeric Approach to Chiral Nonracemic Tricyclic Ring Systems and the Discovery of Dolutegravir (S/GSK1349572) and (S/GSK1265744)

We report herein the discovery of the human immunodeficiency virus type-1 (HIV-1) integrase inhibitors dolutegravir (S/GSK1349572) (<b>3</b>) and S/GSK1265744 (<b>4</b>). These drugs stem from a series of carbamoyl pyridone analogues designed using a two-metal chelation model of the integrase catalytic active site. Structure–activity studies evolved a tricyclic series of carbamoyl pyridines that demonstrated properties indicative of once-daily dosing and superior potency against resistant viral strains. An inherent hemiaminal ring fusion stereocenter within the tricyclic carbamoyl pyridone scaffold led to a critical substrate controlled diastereoselective synthetic strategy whereby chiral information from small readily available amino alcohols was employed to control relative and absolute stereochemistry of the final drug candidates. Modest to extremely high levels of stereochemical control were observed depending on ring size and position of the stereocenter. This approach resulted in the discovery of <b>3</b> and <b>4</b>, which are currently in clinical development