Discovery of HPG1860, a Structurally Novel Nonbile Acid FXR Agonist Currently in Clinical Development for the Treatment of Nonalcoholic Steatohepatitis.

The farnesoid X receptor (FXR) is a ligand-activated nuclear receptor. Activation of FXR significantly impacts the expressions of the pivotal genes involved in bile acid metabolism, inflammation, fibrosis, and homeostasis of lipid and glucose, leading to considerable interests in developing FXR agonists for the treatment of nonalcoholic steatohepatitis (NASH) or other FXR-relevant diseases. Herein, we describe the design, optimization, and characterization of a series of N-methylene-piperazinyl derivatives as the nonbile acid FXR agonists. Particularly, compound 23 (HPG1860), a potent full FXR agonist, shows high selectivity, favorable ADME and pharmacokinetics profile, along with favorable in vivo activities demonstrated in both rodent PD model and HFD-CCl4 model and is currently in clinical development in patients with NASH in phase II.

CircME1 promotes aerobic glycolysis and sunitinib resistance of clear cell renal cell carcinoma through cis-regulation of ME1.

Circular RNAs (circRNAs) play critical roles in clear cell renal cell carcinoma (ccRCC). However, their involvement in sunitinib resistance remains largely unknown. Herein, we identified a novel circRNA, named circME1, which contributes to sunitinib resistance development in ccRCC. CircME1 also promoted proliferation, migration, and invasion of ccRCC cells. Further mechanism analysis showed that circME1 interacted with U1 snRNP at the promoter of its parental gene ME1, thereby upregulating the expression of ME1, enhancing aerobic glycolysis of ccRCC, and promoting its malignant phenotype. Furthermore, ME1 specific inhibitor could effectively repress the oncogenic functions of circME1. Taken together, our study demonstrates that the circME1/ME1 pathway is involved in ccRCC progression and sunitinib resistance development, which may be exploited for anticancer therapy.

Production of functional sperm from in vitro-cultured premeiotic spermatogonia in a marine fish.

In vitro production of functional gametes can revolutionize reproduction by reducing generation intervals and accelerating genetic breeding in aquaculture, especially in fish with relatively long generations. Nevertheless, functional sperm production from in vitro-cultured spermatogonia remains a challenge in most aquaculture fish. In this study, we isolated and characterized premeiotic spermatogonia from marine four-eyed sleepers ( Bostrychus sinensis), which are prone to ovotesticular or sterile testicular development, and induced the differentiation of the spermatogonia into flagellated sperm in a three-dimensional (3D) culture system. Artificial insemination indicated that the in vitro-derived sperm were capable of fertilizing mature oocytes to develop into normal larvae. Furthermore, melatonin significantly promoted spermatogonia proliferation and differentiation through the ERK1/2 signaling pathway, and thus increased the efficiency in functional sperm production. The 3D culture system and resulting functional sperm hold great promise for improving the genetic breeding of aquaculture fish.

Ultrasonic-assisted extraction of polysaccharides from coix seeds: Optimization, purification, and in vitro digestibility.

To optimize the extraction of polysaccharides from coix seeds (CSP), an auxiliary method of ultrasound was developed by response surface methodology (RSM). The maximum extraction yield (8.340%) was obtained under 480 W power, 16 min ultrasound extraction (UE) time and 21.00 mL/g water to raw material ratio. Compared to hot water extraction (HE), UE-treated CSP led to a higher extraction efficiency and decreased average CSP molecular weight. FT-IR indicated that CSP extracted by UE and HE were neutral polysaccharides, and linkages between sugar units were mainly in the α-conformation. Furthermore, NMR spectra indicated that UE-treated CSP was a neutral polysaccharide with (1 â†’ 6)-linked α-d-glucopyranose in the main chain. Two polysaccharide components (CSP-A and CSP-B) were purified by anion exchange chromatography, therein, CSP-A was more resistant to the digestion in stomach and intestine. These results suggest that CSP-A has the potential to be a functional agent utilized by gut microbes.

Identification and Functional Characterization of Metabolites for Bone Mass in Peri- and Postmenopausal Chinese Women.

CONTEXT: Although metabolic profiles appear to play an important role in menopausal bone loss, the functional mechanisms by which metabolites influence bone mineral density (BMD) during menopause are largely unknown. OBJECTIVE: We aimed to systematically identify metabolites associated with BMD variation and their potential functional mechanisms in peri- and postmenopausal women. DESIGN AND METHODS: We performed serum metabolomic profiling and whole-genome sequencing for 517 perimenopausal (16%) and early postmenopausal (84%) women aged 41 to 64 years in this cross-sectional study. Partial least squares regression and general linear regression analysis were applied to identify BMD-associated metabolites, and weighted gene co-expression network analysis was performed to construct co-functional metabolite modules. Furthermore, we performed Mendelian randomization analysis to identify causal relationships between BMD-associated metabolites and BMD variation. Finally, we explored the effects of a novel prominent BMD-associated metabolite on bone metabolism through both in vivo/in vitro experiments. RESULTS: Twenty metabolites and a co-functional metabolite module (consisting of fatty acids) were significantly associated with BMD variation. We found dodecanoic acid (DA), within the identified module causally decreased total hip BMD. Subsequently, the in vivo experiments might support that dietary supplementation with DA could promote bone loss, as well as increase the osteoblast and osteoclast numbers in normal/ovariectomized mice. Dodecanoic acid treatment differentially promoted osteoblast and osteoclast differentiation, especially for osteoclast differentiation at higher concentrations in vitro (eg,10, 100 µM). CONCLUSIONS: This study sheds light on metabolomic profiles associated with postmenopausal osteoporosis risk, highlighting the potential importance of fatty acids, as exemplified by DA, in regulating BMD.

Role of Spinal Cord Akt-mTOR Signaling Pathways in Postoperative Hyperalgesia Induced by Plantar Incision in Mice.

Poor postoperative pain (POP) control increases perioperative morbidity, prolongs hospitalization days, and causes chronic pain. However, the specific mechanism(s) underlying POP is unclear and the identification of optimal perioperative treatment remains elusive. Akt and mammalian target of rapamycin (mTOR) are expressed in the spinal cord, dorsal root ganglion, and sensory axons. In this study, we explored the role of Akt and mTOR in pain-related behaviors induced by plantar incision in mice. Plantar incision activated spinal Akt and mTOR in a dose-dependent manner. Pre-treatment with Akt inhibitors intrathecally prevented the activation of mTOR dose-dependently. In addition, blocking the Akt-mTOR signaling cascade attenuated pain-related behaviors and spinal Fos protein expression induced by plantar incision. Our observations demonstrate that Akt-mTOR might be a potential therapeutic target for the treatment of POP.

Design and Optimization of 3'-(Imidazo[1,2-a]pyrazin-3-yl)-[1,1'-biphenyl]-3-carboxamides as Selective DDR1 Inhibitors.

DDR1 is considered as a promising target for cancer therapy, and selective inhibitors against DDR1 over other kinases may be considered as promising therapeutic agents. Herein, we have identified a series of 3'-(imidazo[1,2-a]pyrazin-3-yl)-[1,1'-biphenyl]-3-carboxamides as novel selective DDR1 inhibitors. Among these, compound 8v potently inhibited DDR1 with an IC50 of 23.8 nM, while it showed less inhibitory activity against DDR2 (IC50 = 1740 nM) and negligible activities against Bcr-Abl (IC50 > 10 µM) and c-Kit (IC50 > 10 µM). 8v also exhibited excellent selectivity in a KINOMEscan screening platform with 468 kinases. This compound dose-dependently suppressed NSCLC cell tumorigenicity, migration, and invasion. Collectively, these studies support its potential application for treatment of NSCLC.

Post-surgical inhibition of phosphatidylinositol 3-kinase attenuates the plantar incision-induced postoperative pain behavior via spinal Akt activation in male mice.

BACKGROUND: Postoperative pain (POP) is a severe acute pain encountered in patients suffering from an operation, and is less than adequately controlled by the currently available analgesics. Phosphatidylinositol 3-kinase (PI3K) has been reported to have an important role in neuropathic and inflammatory pain. Our previous research revealed that pre-surgical inhibition of spinal PI3K alleviated the pain behavior induced by plantar incision in mice. The aim of this study was to clarify whether post-surgical inhibition of PI3K would attenuate the POP and the underlying mechanisms. METHODS: A POP model was established by plantar incision in Kunming mice. A behavioral test was performed to determine mechanical allodynia, thermal hyperalgesia, and cumulative pain scores. The spinal Fos was detected by immunohistochemistry. The spinal expression of protein kinase B (Akt) or phosphorylated Akt (pAkt) was explored using western blot. The cellular location of pAkt was determined by immunofluorescence. RESULTS: Post-surgical inhibition of PI3K attenuated mechanical allodynia, thermal hyperalgesia, and cumulative pain scores induced by plantar incision significantly in male mice, and mildly in female mice. Post-surgical inhibition of PI3K attenuated the expression of spinal Fos in male mice. Plantar incision induced a time-dependent expression of spinal pAkt in male mice, which was primarily expressed in the spinal dorsal horn, and localized with the neuron and microglia's marker. Post-surgical inhibition of PI3K attenuated the activation of Akt induced by plantar incision in male mice as well. CONCLUSIONS: We concluded that post-surgical inhibition of PI3K could attenuate the pain-related behaviors induced by plantar incision, by suppressing the activation of spinal Akt in male mice. This finding might be used in clinical studies to reach a better understanding of POP mechanisms and optimal treatment.

Normalizing JMJD6 Expression in Rat Spinal Dorsal Horn Alleviates Hyperalgesia Following Chronic Constriction Injury.

Jumonji domain-containing protein 6 (JMJD6) is a homolog of hypoxia-inducible factor (HIF) asparaginyl hydroxylase, an inhibitor of HIF. HIF-1α is known to participate in neuropathic pain (NPP) during chronic constriction injury (CCI); however, the roles of JMJD6 in NPP have not been systematically investigated. In this study, we examined the temporal distribution and cellular location of JMJD6 in the spinal cord during CCI. In addition, we assessed behavioral changes representative of NPP in rats. Following CCI, lentiviral vectors (LV-JMJD6) were intrathecally administered to observe the changes in the expression of JMJD6, HIF-1α, and its downstream factor caspase-3. Co-immunoprecipitation was used to detect potential interactions between JMJD6 and HIF-1α. We found that JMJD6 was decreased in rats following CCI, which was accompanied by significant NPP-associated behavioral changes. JMJD6 was mainly expressed in neurons. Intrathecal injection of LV-JMJD6 following CCI alleviated the thermal and mechanical hyperalgesia, normalized JMJD6 protein expression, and decreased HIF-1α protein expression with a corresponding reduction in caspase-3 protein expression. Furthermore, the co-immunoprecipitation analyses showed that JMJD6 and HIF-1α protein immunoprecipitated with each other, indicating an interaction between these two proteins. Taken together, the results suggest that JMJD6 may serve as a sensor in neurons of the adult rat spinal cord during the CCI state. Furthermore, JMJD6 may exert its function in NPP by regulating HIF-1α in rats exposed to CCI.

JMJD6 exerts function in neuropathic pain by regulating NF­κB following peripheral nerve injury in rats.

Treatment of neuropathic pain (NPP) continues to be a major challenge, and the underlying mechanisms remain to be elucidated. Previous studies have demonstrated that histone methylation is important in synaptic plasticity of the nervous system and may affect nuclear factor­κB (NF­κB) signaling through epigenetic mechanisms. The present study aimed to investigate the role of Jumonji C domain 6 (JMJD6), a histone demethylase, in a chronic constriction injury (CCI) model of NPP. On the third day post­CCI surgery, a JMJD6 overexpressing lentiviral vector (LV­JMJD6) was intrathecally injected in the rats. Mechanical withdrawal threshold and thermal withdrawal latency were assessed prior surgery and on days 3, 7, 10 and 14 post­CCI. The results showed that intrathecal injection with the LV­JMJD6 attenuated CCI­induced pain facilitation. The expression of JMJD6 was lower following CCI surgery, and its expression was significantly increased following intrathecal injection with LV­JMJD6, compared with levels in normal saline (NS)­ and negative control lentiviral vector (NC)­treated rats. The expression of spinal NF­κB phosphorylated (p­)p65 subunit and its downstream pain­associated effectors, including interleukin 1ß (IL­1ß), tumor necrosis factor­α (TNF­α) and vascular endothelial growth factor (VEGF), were increased following CCI surgery. Intrathecal injection with LV­JMJD6 suppressed activation of the p­p65 subunit in CCI rats. In addition, expression levels of its downstream effectors IL­1ß, TNF­α and VEGF were attenuated by intrathecal treatment with LV­JMJD6, compared with those in the NS­ and NC­treated CCI rats. Furthermore, the JMJD6­ and p65­immunoreactive cells overlapped in the spinal dorsal horn, however, co­immunoprecipitation showed that JMJD6 and the NF­κB p65 subunit did not directly interact, indicating other functional connections may exist between these factors following CCI surgery. Collectively, these findings indicated an important mechanism underlying the pathogenesis of NPP. JMJD6 may exert its therapeutic function in NPP by regulating NF­κB following CCI.

2-Aminopyrimidine Derivatives as New Selective Fibroblast Growth Factor Receptor 4 (FGFR4) Inhibitors.

A series of 2-aminopyrimidine derivatives were designed and synthesized as highly selective FGFR4 inhibitors. One of the most promising compounds 2n tightly bound FGFR4 with a Kd value of 3.3 nM and potently inhibited its enzymatic activity with an IC50 value of 2.6 nM, but completely spared FGFR1/2/3. The compound selectively suppressed proliferation of breast cancer cells harboring dysregulated FGFR4 signaling with an IC50 value of 0.38 µM. Furthermore, 2n exhibited extraordinary target specificity in a Kinome-wide screen against 468 kinases, with S(35) and S(10) selectivity scores of 0.01 and 0.007 at 1.0 µM, respectively.

Structure-Based Design of Tetrahydroisoquinoline-7-carboxamides as Selective Discoidin Domain Receptor 1 (DDR1) Inhibitors.

The structure-based design of 1, 2, 3, 4-tetrahydroisoquinoline derivatives as selective DDR1 inhibitors is reported. One of the representative compounds, 6j, binds to DDR1 with a Kd value of 4.7 nM and suppresses its kinase activity with an IC50 value of 9.4 nM, but it is significantly less potent for a panel of 400 nonmutated kinases. 6j also demonstrated reasonable pharmacokinetic properties and a promising oral therapeutic effect in a bleomycin-induced mouse pulmonary fibrosis model.

Matrix metalloproteinase-9 is required for vasculogenic mimicry by clear cell renal carcinoma cells.

BACKGROUND: Vasculogenic mimicry (VM), a new pattern of tumor microcirculation system, has been proved to be important for tumor growth and progression and may be one of the causes of antiangiogenesis resistance. Matrix metalloproteinase-9 (MMP9) was shown to correlate with VM formation in some other cancers. However, the relationship between VM formation and MMP9 in renal cell carcinoma (RCC) has not been determined. METHODS: The VM formation and MMP9 expressions were analyzed by CD34/periodic acid-Schiff dual staining and immunohistochemistry in 119 RCC specimens. We used a well-established 3-dimention culture model to compare VM formation in 786-O, 769-P, and HK-2 cell lines in vitro. MMP9 expressions on either messenger RNA or protein levels were compared among the cell lines by quantitative polymerase chain reaction or Western blot. To determine further the relationship between MMP9 and VM in RCC, 786-O and 769-P were treated with specific MMP9 inhibitor or small interfering RNA. VM formation, cell migration, and invasion were subsequently assessed by 3-dimention culture, wound-healing, and transwell assays. RESULTS: Immunohistochemistry demonstrated both VM formation and MMP9 overexpression were positively associated with clinical staging, pathological grade, and metastasis (P<0.01). VM formation was closely correlated with MMP9 overexpression in RCC (r = 0.602, P<0.01). Lower MMP9 expression level was observed in normal kidney cell line HK-2, which was unable to form VM on Matrigel, whereas higher expression of MMP9 was found in VM-forming cancer cell lines 786-O and 769-P. Inhibition of MMP9 not only disrupted VM formation in 786-O and 769-P but also reduced cell migration and invasion. CONCLUSIONS: These results indicate an intimate relationship between MMP9 overexpression and VM formation in RCC. Treatments targeting VM formation by inhibiting the activity of MMP9 could be beneficial in RCC therapy.

Comparison between laparoscopic partial nephrectomy and laparoscopic ablation therapy: a meta-analysis.

OBJECTIVE: To conduct a meta-analysis of the literature evaluating comparisons on the peri-operative and oncological outcomes between laparoscopic partial nephrectomy (LPN) and laparoscopic ablation therapy (LAT) in the treatment of small renal masses (SRMs). MATERIAL AND METHODS: MEDLINE, EMBASE, Google Scholar, Cochrane Library, and CNKI were searched for clinical trials comparing LPN with LAT. Data of peri-operative and follow-up outcomes were extracted and compared. Publication bias was identified and sensitivity analysis was also performed. RESULTS: Data from 11 studies including 928 patients (525 patients in the LPN group and 403 in the LAT group) were collected. Baseline characteristics were compared and differences were found in age, preoperative renal function and proportion of solitary kidney (p < 0.05 respectively). For peri-operative outcomes, the LPN group had greater estimated blood loss, longer operative duration and length of hospital stay, and more peri-operative complications (p < 0.05, respectively). The LAT group had a significantly higher local recurrence (p < 0.05). There was no significant difference in postoperative change of renal function (p = 0.21). CONCLUSION: In comparison with LPN, LAT provides better peri-operative outcomes, but a higher local recurrence rate. LAT does not seem to provide an obvious advantage in protecting renal function. Further clinical trials with randomized design and long-term follow-up are needed.

[Protective effects of bicyclol on alcohol-induced liver damage in mice].

OBJECTIVE: To study the protective effect of bicyclol on alcohol-induced liver injury in mice. METHODS: Sixty male mice were randomly divided into 6 groups. Ten mice were fed with Lieber-Decarli liquid diet without alcohol and used as normal controls. Fifty mice were fed with Lieber-Decarli liquid diet containing 5% alcohol for four weeks so as to establish a model of alcohol-induced liver damage. Ten mice fed with Lieber-Decarli liquid diet containing 5% alcohol for four weeks were used as model group. Bicyclol in a dose of either 200 or 300 .kg(-1).d(-1) was given orally simultaneously with alcohol intake as prevention groups (10 mice in each group), and bicyclol in a dose of either 200 or 300 .kg(-1).d(-1) was given orally 2 weeks after the beginning of alcohol intake as treatment groups (10 mice in each group). Twenty-four hours after the last dose of bicyclol the mice were decapitated and then their blood samples and livers were taken. The serum alanine aminotransferase (ALT), cholesterol (CHOL), and high-density lipoprotein/low-density lipoprotein (HDL/LDL), and liver triglyceride (TG), N-nitrosodimethylamine demethylase (NDMA-DM), glutathione (GSH), glutathione S-transferase (GST), glutathione reductase (GR), and aldehyde dehydrogenase (ALDH) were determined by biochemical assays. The extent of liver damage was evaluated by histological examination. RESULTS: Four weeks after alcohol intake the serum ALT and TG were 1.9 and 2.7 times those of the normal control group. The levels of liver TG of the bicyclol 200 kg(-1).d(-1) and 300 kg(-1).d(-1) treatment groups were significantly lower than that of the model group by 28% and 32% respectively (both P < 0.05). The levels of liver TG of the bicyclol 200 kg(-1).d(-1) and 300 kg(-1).d(-1) prevention groups were significantly lower than that of the model group by 32%, and 47% respectively (both P < 0.01). Pathological changes including steatosis and hepatocyte ballooning degeneration were found in the livers of the model group. The levels of liver GSH, GST, and GR in the model group decreased by 37%, 22%, and 19% in comparison with the normal control group. The levels of liver GSH and GST of the bicyclol prevention groups were normal, and the liver GR level was 1.2 times that of the normal control group. The liver NDMA-DM activity of the model group was 1.9 times that of the normal control group and was normal in the bicyclol prevention and treatment groups. The liver cytoplasmic ALDH level was 30% lower in the model group than in the normal control group (P < 0.05), and was 2.9 times in the bicyclol groups (P < 0.01). The serum cholesterol levels of the bicyclol groups were all significantly lower than that of the model group (all P < 0.01). The serum levels of HDL of the bicyclol prevention groups and treatment were all significantly lower than that in the model group (P < 0.01 or P < 0.05). CONCLUSION: Bicyclol protects mice against alcohol-induced hepatotoxicity by reduction of hepatic steatosis and cellular damage, acceleration of alcohol and aldehyde elimination and anti-peroxidation.