Chemical Synthesis, Safety and Efficacy of Antihypertensive Candidate Drug 221s (2,9).

Hypertension is the main risk factor of cardiovascular and cerebrovascular diseases. In this paper, a novel compound known as 221s (2,9), which includes tanshinol, borneol and a mother nucleus of ACEI, was synthesized by condensation esterification, deprotection, amidation, deprotection, and amidation, with borneol as the initial raw material, using the strategy of combinatorial molecular chemistry. The structure of the compound was confirmed by 1H NMR, 13C NMR, and high-resolution mass spectrometry, with a purity of more than 99.5%. The compound 221s (2,9) can significantly reduce the systolic and diastolic blood pressure of SHR rats by about 50 mmHg and 35 mmHg after 4 weeks of administration. The antihypertensive effect of 221s (2,9) is equivalent to that of captopril. The use of 221s (2,9) can reduce the content of Ren, Ang II and ACE in the serum of SHR rats, inhibit the RAAS and enhance the vascular endothelial function by upregulating the level of NO. Pathological studies in this area have shown that high dosage of 221s (2,9) can notably protect myocardial fibrosis in rats and reduce the degeneration and necrosis of myocardial fibers, inflammatory cell infiltration, and proliferation of fibrous tissue in the heart of rat. Therefore, the existing work provided a foundation for preclinical research and follow-up clinical research of 221s (2,9) as a new drug.

Boronate affinity magnetic nanoparticles with hyperbranched polymer brushes for the adsorption of cis-diol biomolecules.

A boronate-modified magnetic affinity sorbent was prepared by adopting hyperbranched polyethyleneimine as the scaffold to amplify initiator sites. 3-Acrylamidophenylboronic acid was employed as monomer to proceed in situ free-radical polymerization on magnetite (Fe3O4) nanoparticles. Due to the improved density of boronic acid polymers, the adsorbent exhibited high adsorption capacity, typically (134 ± 8) µg mg-1 for dopamine, (92 ± 7) µg mg-1 for catechol, (363 ± 14) µg mg-1 for ovalbumin and (464 ± 22) µg mg-1 for horseradish peroxidase. These capacities are much higher than those of other adsorbents. The sorbent was applied to the enrichment of catecholamines from spiked human urine. Owing to the high adsorption capacity, only 1.0 mg of adsorbent was sufficient to eliminate the interferences and enrich the targets (dopamine, norepinephrine and epinephrine) within 5 min. They were quantified by HPLC. The recoveries from spiked samples range between 83.5% ~106%, with relative standard deviations of 3.2 ~ 9.4% (n = 5). The separation of glycoproteins from egg white was also accomplished prior to their analysis by PAGE. In the authors' perception, this approach is promising in that the density of functional groups on the adsorbent is strongly increased. Graphical abstract The preparation routine of boronate affinity magnetic adsorbent (Fe3O4@HpAAPBA). The adsorbent is used for the magnetic solid phase extraction of cis-dol compounds from real sample.

Facile preparation of boronic acid-functionalized magnetic nanoparticles with a high capacity and their use in the enrichment of cis-diol-containing compounds from plasma.

The use of bronate affinity adsorbents is a new separation method that appeared recently with great potential for specific extraction of cis-diol-containing compounds. In this work,a new strategy for the facile construction of boronic acid-functionalized Fe3 O4 magnetic nanoparticles (Fe3 O4 @FPBA MNPs) with a high capacity was described. The extraction capacity of the Fe3 O4 @FPBA MNPs was determined to be 66.0 ± 2.7 µmol/g for catechol and 80.6 ± 2.0 µmol/g for dopamine, being higher than that for the reported methods. The Fe3 O4 @FPBA MNPs were used to extract four cis-diol drugs: caffeic acid isopropyl ester, caffic acid bornyl ester, isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate and 3-(3, 4-dihydroxyphenyl)-2-hydroxylpropionic acid - from the spiked rabbit plasma, and the recoveries of four drugs were between 87.29 and104.37% with relative standard deviations ranging from 1.34 to 8.81%. Under the most favorable conditions, the solid-phase extraction combined with HPLC-UV for the analysis of four drugs in plasma could eliminate interferences from endogenous components of the biological fluids and exhibited sufficient precision and accuracy. These results showed that the prepared Fe3 O4 @FPBA MNPs were qualified for efficiently enriching and determining the trace cis-diol substances from biological samples.

[On-line enrichment and separation of glycoprotein by capillary electrophoresis based on pH response coating column].

A capillary column coated with 3-aminophenylboronic acid (APBA)-functionalized gold nanoparticles (AuNPs@APBA) was prepared via electrostatic self-assembly. The coated column exhibited anti-nonspecific adsorption of glycoproteins, enabling selective online enrichment during capillary electrophoresis (CE). First, gold nanoparticles (AuNPs) were synthesized using the sodium citrate reduction method. Then, APBA was self-assembled electrostatically on the surface of the AuNPs to obtain AuNPs@APBA. This nanomaterial was bonded to the inner wall of a capillary through ion adsorption to produce a AuNPs@APBA-coated capillary column. Glycoproteins were adsorbed via bond formation with boric acid groups under alkaline conditions (pH 8) to generate borate esters. Under acidic conditions (pH 3), the borate esters dissociated to release the glycoproteins, thereby achieving the selective online enrichment and separation of glycoproteins. The AuNPs and AuNPs@APBA were characterized using Fourier transform infrared spectroscopy, and their sizes and Zeta potentials were determined. In addition, the electroosmotic flow (EOF) of the AuNPs@APBA-coated capillary column was measured. The results showed that the surface of the AuNPs was successfully modified with APBA and that AuNPs@APBA was adsorbed on the inner wall of the capillary. The peak area of ovalbumin (OVA) on the AuNPs@APBA-coated column was 26.46 times higher than that on a bare column via conventional electrophoresis. In contrast, the peak area of bovine serum albumin (BSA) only increased by 8.47 times, indicating that the AuNPs@APBA coated column selectively enriched glycoproteins. Evaluation of the reproducibility and stability of this method revealed that the AuNPs@APBA coated capillary column could be used continually for 33-67 h. The relative standard deviations (RSDs) of the peak areas for intra-day (n=5) and inter-day (n=6) analyses were 2.2% and 3.0%, respectively. The developed method was successfully applied to enrich glycoproteins in a 1×106-fold diluted egg white sample. Glycoproteins were not detected using conventional electrophoresis on the bare column, whereas the AuNPs@APBA-coated capillary column effectively enriched and separated glycoproteins, resulting in a peak area of 10469 mAU·ms. Furthermore, the entire enrichment and separation process was completed within 3 min. This new online enrichment and separation method for glycoproteins has the advantages of low sample consumption, simple operation, and high separation efficiency.

LncRNA NORAD accelerates the progression of non-small cell lung cancer via targeting miRNA-455/CDK14 axis.

BACKGROUND: To explore the potential involvement of long non-coding RNA (lncRNA) NORAD in regulating the progression of Non-small cell lung cancer (NSCLC), and its possible mechanism. METHODS: Relative level of NORAD in NSCLC tissues and cell lines was determined. Its level in NSCLC patients with different tumor staging (T1-T2, T3-T4) and either with lymphatic metastasis or not was examined as well. Kaplan-Meier curves were depicted for assessing the prognostic value of NORAD in NSCLC. Regulatory effects of NORAD on the proliferative ability of NCI-H1650 and HCC827 cells were evaluated. Dual-luciferase reporter gene assay was conducted to identify the binding between NORAD and miRNA-455, as well as between miRNA-455 and CDK14. At last, the role of NORAD/miRNA-455/CDK14 regulatory loop in influencing the progression of NSCLC was determined. RESULTS: NORAD was upregulated in NSCLC tissues and cells. Its level was higher in NSCLC patients with advanced stage or accompanied with lymphatic metastasis. Worse prognosis was observed in NSCLC patients presenting high level of NORAD. Silence of NORAD attenuated the proliferative ability of NCI-H1650 and HCC827 cells. MiRNA-455 was the downstream target binding to NORAD. Its level was negatively regulated by NORAD. Knockdown of miRNA-455 could reverse the role of NORAD in regulating the proliferative ability of NSCLC. Moreover, CDK14 was the target gene of miRNA-455. CDK14 level was negatively regulated by miRNA-455. CONCLUSIONS: LncRNA NORAD is upregulated in NSCLC, which enhances the proliferative ability of tumor cells by targeting miRNA-455/CDK14 axis and thereby accelerates the progression of NSCLC.

[Determination of gastrodin activity inhibition on acetylcholinesterase by capillary electrophoresis].

Alzheimer's disease (AD) is the most common cause of dementia in elderly individuals. Currently, acetylcholinesterase inhibitors (AChEI) are the most effective clinical treatment for AD. AChEIs in natural products may have therapeutic potential and should be screened for use in AD treatment. The authors describe a simple and reliable method for AChEI screening by capillary electrophoresis (CE). A hexadimethrine bromide (HDB) solution was pushed into a capillary (0.015 MPa×10 s) and incubated for 5 min. The capillary was flushed with deionized water for 5 min to remove free HDB, followed by plugging with an acetylcholinesterase (AChE) solution. After a 5 min incubation, the AChE was immobilized on the positively charged coating by ion binding, and the micro-reactor was created. The substrate solution, acetylthiocholine iodide (AThC), was injected into the capillary and incubated in the micro-reactor for 1 min. The unreacted substrate and the enzymolysis product were separated by CE. Gastrodin, an important component of Gastrodia elata, can inhibit AChE activity. After a certain amount of gastrodin was spiked into the substance solution, the peak area of the product decreased. Greater peak area reduction indicated stronger inhibition of AChEI. We observed good reproducibility of the product peak, with relative standard deviation (RSD) values less than 5.3%. The micro-reactor can be reused up to 300 times, which greatly improves efficiency. When the concentration of gastrodin was 5.24 µmol/L, the inhibition rate of AChE reached 64.8%. The IC50 of gastrodin was (2.26±0.14) µmol/L (R2=0.9983), which was consistent with the result of traditional UV method (2.09±0.18 µmol/L). If the function of the micro-reactor deteriorates, it can be conveniently renewed by flushing the column to remove the enzyme and repeating the AChE immobilization protocol. The proposed method is simple, efficient, and low cost, and can be used to screen AChEI from natural products, thus contributing to the improvement of AD treatment.

Specific enrichment of caffeic acid from Taraxacum mon-golicum Hand.-Mazz. by pH and magnetic dual-responsive molecularly imprinted polymers.

Specific recognition of caffeic acid (CA) from Taraxacum mon-golicum Hand.-Mazz. was successfully performed using a new pH responsive magnetic molecularly imprinted polymers (pH-MMIPs) by simple surface molecular imprinting polymerization. The pH-MMIPs were prepared on the surface of the Fe3O4@SiO2@MPS particles using CA as a template, 2-(dimethylamino) ethyl methacrylate (DMA) as the pH responsive functional monomer, 4-vinylpyridine (4-VP) as an assisting functional monomer, ethylene glycol dimethyl acrylate (EGDMA) as cross-linker, 2,2'-azobisisobutyronitrile (AIBN) as initiator and methanol-H2O (1:1, v/v) as the porogen. The resultant polymers were characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), vibrating sample magnetometry (VSM) and x-ray diffraction (XRD). The adsorption experiments revealed that the pH-MMIPs performed high adsorption ability (11.5 mg g-1) by changing solution pH. Successful selective adsorption of CA was achieved with distribution coefficient of 0.12 and 0.21 towards ferulic acid and chlorogenic acid. Furthermore, pH-MMIPs were employed as adsorbents for extraction and enrichment of CA from Taraxacum mon-golicum Hand.-Mazz. extract. The recoveries of CA in the Taraxacum mon-golicum Hand.-Mazz. ranged from 90.47% to 98.97%. The results proved that the polymers have the potential to provide a selective recognition of CA in complex samples by simple pH regulation.

Multidentate boronate magnetic adsorbent assembled with polyhedral oligomeric silsesquioxanes and intramolecular diboronic acid for improving the binding strength toward glycoproteins.

Boronate affinity is an important method for the enrichment and separation of cis-diol containing compounds, but most of the conventional boronate materials suffer from weak binding strength as well as low binding capacity towards glycoproteins due to the use of single boronic acids as ligands. In this work, a novel multidentate boronate magnetic adsorbent was assembled by using amined polyhedral oligomeric silsesquioxane as spacer and a diboronic acid as ligand. The specially designed adsorbent exhibited high adsorption capacity for cis-diols due to the high density of phenylbronic acid moieties. More interestingly, the dissociation constants toward glycoproteins on the material were lowered to be ∼10-6 M, being at least 3 orders lower than the single boronic acid bonded adsorbents. By comparing the binding properties of small molecules containing one and two pairs of cis-diols, the enhanced binding strength of glycoproteins on the multidentate boronate magnetic adsorbent was attributed to the synergistic binding of glycoproteins on the special interface. The new materials successfully captured glycoproteins from 1000-fold diluted egg white, suggesting that the material could be an optional alternative adsorbent for enriching trace glycoproteins from complex bio-samples.

Selective enrichment and determination of monoamine neurotransmitters by CU(II) immobilized magnetic solid phase extraction coupled with high-performance liquid chromatography-fluorescence detection.

In this paper, iminodiacetic acid-Cu(II) functionalized Fe3O4@SiO2 magnetic nanoparticles were prepared and used as new adsorbents for magnetic solid phase extraction (MSPE) of six monoamine neurotransmitters (MNTs) from rabbit plasma. The selective enrichment of MNTs at pH 5.0 was motivated by the specific coordination interaction between amino groups of MNTs and the immobilized Cu(II). The employed weak acidic extraction condition avoided the oxidation of MNTs, and thus facilitated operation and ensured higher recoveries. Under optimal conditions, the recoveries of six MNTs from rabbit plasma were in the range of 83.9-109.4%, with RSD of 2.0-10.0%. When coupled the Cu(II) immobilized MSPE with high-performance liquid chromatography-fluorescence detection, the method exhibited relatively lower detection limits than the previously reported methods, and the method was successfully used to determine the endogenous MNTs in rabbit plasma. The proposed method has potential application for the determination of MNTs in biological samples. Also, the utilization of coordination interaction to improve the selectivity might open another way to selectively enrich small alkaloids from complex samples.

Preparation of a novel Zr(4+)-immobilized metal affinity membrane for selective adsorption of phosphoprotein.

In this study, a novel phosphate-Zr(4+) immobilized metal affinity membrane (IMAM) was prepared based on the surface initiated-atom transfer radical polymerization technique for the selective adsorption of phosphoprotein. The adsorption capacity and selectivity of the phosphate-Zr(4+) IMAM were evaluated by using the mixture of standard phosphoproteins (ß-casein, ovalbumin) and nonphosphoproteins (bovine serum albumin and lysozyme) as model samples. The adsorption isotherms and competitive adsorption results demonstrated that the phosphate-Zr(4+) IMAM had higher binding capacity and selectivity for phosphoproteins over nonphosphoproteins. Moreover, the phosphate-Zr(4+) IMAM exhibited good re-usability and re-productivity. Finally, the phosphate-Zr(4+) IMAM was applied to separate phosphoprotein from real samples with high purity. Therefore, the as-prepared phosphate-Zr(4+) IMAM could be a promising affinity material for the efficient enrichment of phosphoprotein from complex bio-samples.

Enhanced binding capacity of boronate affinity adsorbent via surface modification of silica by combination of atom transfer radical polymerization and chain-end functionalization for high-efficiency enrichment of cis-diol molecules.

Boronate affinity materials have been widely used for specific separation and preconcentration of cis-diol molecules, but most do not have sufficient capacity due to limited binding sites on the material surface. In this work, we prepared a phenylboronic acid-functionalized adsorbent with a high binding capacity via the combination of surface-initiated atom transfer radical polymerization (SI-ATRP) and chain-end functionalization. With this method, the terminal chlorides of the polymer chains were used fully, and the proposed adsorbent contains dense boronic acid polymers chain with boronic acid on the chain end. Consequently, the proposed adsorbent possesses excellent selectivity and a high binding capacity of 513.6 µmol g(-1) for catechol and 736.8 µmol g(-1) for fructose, which are much higher than those of other reported adsorbents. The dispersed solid-phase extraction (dSPE) based on the prepared adsorbent was used for extraction of three cis-diol drugs (i.e., epinephrine, isoprenaline and caffeic acid isopropyl ester) from plasma; the eluates were analyzed by HPLC-UV. The reduced amount of adsorbent (i.e., 2.0 mg) could still eliminate interferences efficiently and yielded a recovery range of 85.6-101.1% with relative standard deviations ranging from 2.5 to 9.7% (n = 5). The results indicated that the proposed strategy could serve as a promising alternative to increase the density of surface functional groups on the adsorbent; thus, the prepared adsorbent has the potential to effectively enrich cis-diol substances in real samples.