Photoreactive heparin was synthesized by reaction of 4-azidoaniline and heparin. An organic layer was introduced on the surface of Ti-O by 3-aminopropylphosphonic acid assembling, and then the modified heparin was immobilized on the surface by UV irradiation. Water contact angle was used to characterize the hydrophilicity, quantitive assay was done by azure staining methods, and blood compatibility was evaluated by platelet adhesion experiment. Water contact angle of heparinized surface was smaller than that of Ti-O film, which indicated more hydrophilic property of heparinized surface. The surface density of heparin increased with the prolonging of irradiation time and the density was 2.1 microg/cm2 when irradiated for 300s. It showed the heparinized surface was effective in resisting platelets from adhesion and aggregation.
Aniline Compounds ; chemistry ; Azo Compounds ; chemistry ; Blood Proteins ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Heparin ; chemistry ; Humans ; Membranes, Artificial ; Propylamines ; chemistry ; Surface Properties ; Titanium ; chemistry

OBJECTIVETo study whether the analgesis of oxymatrine (OMT) affects N-type voltage-gated calcium channels (VGCCs).

METHODSTotally 45 mice were randomly divided into the sham-operation group, the model group [established by partial sciatic nerve ligation (PSNL)] , and the OMT treatment group according to random digit table, 15 in each group. The dorsal root ganglions (DRG) were separated in PSNL pain model mice. Intracellular calcium concentration ([Ca2+]i) was determined with Fluo-3 AM immunofluorescent probe in cultured DRG neurons. Different protein expression levels of N-type (Cav2. 2) and L-type ( Cav1. 3) among VGCCs from brain and DRG tissues were detected with Western blot.

RESULTSCompared with the sham-operation group, [Ca2+]i, increased in cultured DRG neurons (P <0. 05) , protein expression levels of Cav2. 2 in the brain tissue increased (P <0. 05), protein expression levels of Cav2. 2 in DRG tissues decreased in the model group (P <0. 01). Compared with the model group, [Ca2+]i, decreased in cultured DRG neurons (P < 0. 05), protein expression levels of Cav2. 2 in the brain tissue decreased (P <0. 01), protein expression levels of Cav2. 2 in DRG tissues increased in the OMT treatment group (P <0. 01). There was no statistical difference in Cav1. 3 expressions in cultured DRG neurons and the brain (P >0. 05).

CONCLUSIONAnalgesic effect of OMT might be related to Cav2. 2 channel mediated calcium ion flux.

Alkaloids ; pharmacology ; Analgesia ; methods ; Analgesics ; pharmacology ; Aniline Compounds ; Animals ; Calcium ; Calcium Channels, N-Type ; physiology ; Ganglia, Spinal ; Mice ; Neurons ; Pain ; Quinolizines ; pharmacology ; Xanthenes

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.
Aniline Compounds ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Humans ; Inhibitory Concentration 50 ; Phosphorylation ; Quinazolines ; pharmacology ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors

A series of N-(3-cyano-6-methyl-4-anilinoquinoline-7-yl) amide derivatives 13a-13n have been synthesized, their structures were confirmed with 1H NMR, EI-MS, IR and elemental analysis. Antitumor activities of all the synthesized compounds in vitro were tested with MTT. Compound 13j showed better than or equal antitumor activity on Bosutinib and EKB-569.
Aniline Compounds ; chemical synthesis ; chemistry ; pharmacology ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Humans ; Quinolines ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo identify the bacteria tolerating chlorinated anilines and to study the biodegradability of o-chloroaniline and its coexistent compounds.

METHODSMicrobial community of complex bacteria was identified by plate culture observation techniques and Gram stain method. Bacterial growth inhibition test was used to determine t he tolerance of complex bacteria to toxicant. Biodegradability of chlorinated anilines was determined using domesticated complex bacteria as an inoculum by shaking-flask test.

RESULTSThe complex bacteria were identified, consisting of Xanthomonas, Bacillus alcaligenes, Acinetobacter Pseudomonas, and Actinomycetaceae nocardia. The obtained complex bacteria were more tolerant to o-chloroaniline than mixture bacteria in natural river waters. The effects of exposure concentration and inoculum size on the biodegradability of o-chloroaniline were analyzed, and the biodegradation characteristics of single o-chloroaniline and 2, 4-dichloroaniline were compared with the coexistent compounds.

CONCLUSIONThe biodegradation rates can be improved by decreasing concentration of compounds and increasing inoculum size of complex bacteria. When o-chloroaniline coexists with aniline, the latter is biodegraded prior to the former, and as a consequence the metabolic efficiency of o-chloroaniline is improved with the increase of aniline concentration. Meanwhile, when o-chloroaniline coexists with 2,4-dichloroaniline, the metabolic efficiency of 2,4-dichloroaniline is markedly improved.

Aniline Compounds ; metabolism ; pharmacology ; Bacteria ; drug effects ; growth & development ; metabolism ; Biodegradation, Environmental ; Rivers ; chemistry ; microbiology ; Water Pollutants, Chemical ; metabolism ; pharmacology ; Water Purification

OBJECTIVETo observe the role of NB127914, a CRF R1 receptor antagonist, in the regulation of neonatal sleep/wake cycle.

METHODSRat pups were surgically implanted with electrodes at postnatal day(PN) 13. At PN 14, 6 hours polysomnographic recording data were continuously collected before and after administration of various doses of NBI 27914, atropine and the same amount of saline.

RESULTSCompared with baseline, rapid eye movement (REM) sleep was significantly reduced and was replaced primarily by non-REM (NREM) sleep in all groups treated with NBI, but not with dimethyl sulfoxide/saline. Atropine suppressed REM sleep significantly and increased wakefulness simultaneously.

CONCLUSIONBlockage of corticotropin-releasing factor (CRF) R1 receptors deprives neonatal rat REM sleep.

Aniline Compounds ; pharmacology ; Animals ; Female ; Male ; Polysomnography ; Pyrimidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Corticotropin-Releasing Hormone ; antagonists & inhibitors ; Sleep, REM ; drug effects ; physiology ; Wakefulness ; drug effects ; physiology

The discovery of high performance leading antidiabetic compounds containing sulfonamide and 4-aminophenylacetic acid moieties is reported. This was achieved by the synthesis of 6 intermediates and subsequently 20 target molecules using 4-aminophenylacetic acid as the starting materials, and through a few synthetic routes aided by multi-step reactions including sulfonylation of amino group, deacylation of amides and esterification of carboxyl group, as well as acylation of amino group. The chemical structures of the twenty-four new compounds were determined using 1H NMR, 13C NMR and HR-MS techniques. Screening in vitro of their peroxisome proliferator-activated receptor (PPAR) activation activities showed weak relative PPAR activation activities to most of the target molecules. However, 4 target molecules exhibit PPAR over 58%, and as high as 81.79% for TM2-i, presenting itself as potent leading compound for antidiabetic drugs. This research also confirms that it is probable to achieve esterification of carboxyl group and deacylation of fatty acid N-phenyl amides concurrently in SOCl2/alcohol solvent system. This provides new synthetic method for the selective reaction within molecules containing both carboxyl and N-aryl amido groups of fatty acids.
Aniline Compounds ; chemistry ; Fatty Acids ; chemistry ; Hep G2 Cells ; metabolism ; Humans ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; Peroxisome Proliferator-Activated Receptors ; metabolism ; Phenylacetates ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Sulfonamides ; chemistry

Epidermal growth factor receptor (EGFR)-targeted therapies have been effective in some cancers, but not in hepatocellular carcinoma (HCC). The aim of this study was to investigate the drug potential to overcome multi-drug resistance in HCC cells. Thirteen drug-sensitive HCC cells were assessed using the CCK-8 assay. G0-G1 arrest was measured by FACS. Western blot analysis was used to detect the key enzymes in both the Ras/Raf and PI3K pathways. When establishing the IC50 of HCC to several drugs, including EKB-569, sorafenib, erlotinib, gefitinib, pazopanib, and brivanib, SK-Hep1 cells treated with EKB-569 have shown the highest (72.8%-86.4%) G0-G1 arrest and decreased the phosphorylation of AKT and ERK at the protein level. We found that EKB-569 had higher efficacy in HCC, compared to first generation, reversible EGFR-TK inhibitors. Furthermore, the combination of sorafenib and EKB-569 showed a synergistic effect to inhibit proliferation of SNU-475, previously the most resistant cell to EGFR-TKIs. Therefore, novel EKB-569 in combination with sorafenib may be able to overcome HCC resistance to EGFR-TK inhibitors.
Aminoquinolines/*pharmacology ; Aniline Compounds/*pharmacology ; Antineoplastic Agents/*pharmacology ; Antineoplastic Combined Chemotherapy Protocols/pharmacology ; Benzenesulfonates/pharmacology ; Carcinoma, Hepatocellular/*drug therapy/pathology ; Cell Cycle Checkpoints/drug effects ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Liver Neoplasms/drug therapy/pathology ; Mitogen-Activated Protein Kinases/*metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt/*metabolism ; Pyridines/pharmacology ; Receptor, Epidermal Growth Factor/*antagonists & inhibitors

Voltage-gated sodium channels (VGSCs) in primary sensory neurons play a key role in transmitting pain signals to the central nervous system. BmK I, a site-3 sodium channel-specific toxin from scorpion Buthus martensi Karsch, induces pain behaviors in rats. However, the subtypes of VGSCs targeted by BmK I were not entirely clear. We therefore investigated the effects of BmK I on the current amplitude, gating and kinetic properties of Nav1.8, which is associated with neuronal hyperexcitability in DRG neurons. It was found that BmK I dose-dependently increased Nav1.8 current in small-sized (<25 μm) acutely dissociated DRG neurons, which correlated with its inhibition on both fast and slow inactivation. Moreover, voltage-dependent activation and steady-state inactivation curves of Nav1.8 were shifted in a hyperpolarized direction. Thus, BmK I reduced the threshold of neuronal excitability and increased action potential firing in DRG neurons. In conclusion, our data clearly demonstrated that BmK I modulated Nav1.8 remarkably, suggesting BmK I as a valuable probe for studying Nav1.8. And Nav1.8 is an important target related to BmK I-evoked pain.
Aniline Compounds ; pharmacology ; Animals ; Cell Size ; Cells, Cultured ; Electrophysiological Phenomena ; drug effects ; Furans ; pharmacology ; Ganglia, Spinal ; cytology ; Kinetics ; Male ; NAV1.8 Voltage-Gated Sodium Channel ; metabolism ; Rats ; Rats, Sprague-Dawley ; Scorpion Venoms ; antagonists & inhibitors ; pharmacology ; Scorpions ; Sensory Receptor Cells ; drug effects ; metabolism ; physiology ; Sodium Channel Blockers ; pharmacology ; Voltage-Gated Sodium Channel Agonists ; pharmacology

OBJECTIVETo investigate the effect of the metabolite of leflunomide, A771726,on proliferation and collagen synthesis of hepatic stellate cell (HSC).

METHODSHSC and Kupffer cells were isolated from the rat liver by collagenase IV and pronase perfusion, and purified by density gradient separation. The effects of A771726 on cell proliferation and collagen synthesis were examined by 3H-thymidine and 3H-proline incorporation assays, respectively. The TGF-beta, TNF-alpha and IL-1 levels in Kupffer cell conditioned medium (KCCM) were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSHSC and Kupffer cells in rat liver were well separated. The KCCM of CCl4-injured rats had significant stimulation effect on proliferation and collagen synthesis of HSC in primary culture. Addition of A771726 (0.001-10 micromol/Lein HSC culture stimulated by KCCM significantly inhibited proliferation and collagen synthesis of HSC. Furthermore, the elevated TGF-beta, TNF-alpha and IL-1 levels in KCCM of CCl4-injured rats were significantly reduced in A771726 treatment groups.

CONCLUSIONA771726 has markedly inhibitory effect on proliferation and collagen synthesis of HSC and secretion of TGF-beta,TNF-alpha and IL-1 from Kupffer cells.

Aniline Compounds ; pharmacology ; Animals ; Carbon Tetrachloride ; Cell Division ; drug effects ; Cells, Cultured ; Collagen Type I ; biosynthesis ; Hepatocytes ; cytology ; Hydroxybutyrates ; pharmacology ; Immunosuppressive Agents ; pharmacology ; Interleukin-1 ; metabolism ; Isoxazoles ; metabolism ; pharmacology ; Kupffer Cells ; cytology ; Male ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; metabolism