Trilogy of drug repurposing for developing cancer and chemotherapy-induced heart failure co-therapy agent

Chemotherapy-induced complications, particularly lethal cardiovascular diseases, pose significant challenges for cancer survivors. The intertwined adverse effects, brought by cancer and its complication, further complicate anticancer therapy and lead to diminished clinical outcomes. Simple supplementation of cardioprotective agents falls short in addressing these challenges. Developing bi-functional co-therapy agents provided another potential solution to consolidate the chemotherapy and reduce cardiac events simultaneously. Drug repurposing was naturally endowed with co-therapeutic potential of two indications, implying a unique chance in the development of bi-functional agents. Herein, we further proposed a novel "trilogy of drug repurposing" strategy that comprises function-based, target-focused, and scaffold-driven repurposing approaches, aiming to systematically elucidate the advantages of repurposed drugs in rationally developing bi-functional agent. Through function-based repurposing, a cardioprotective agent, carvedilol (CAR), was identified as a potential neddylation inhibitor to suppress lung cancer growth. Employing target-focused SAR studies and scaffold-driven drug design, we synthesized 44 CAR derivatives to achieve a balance between anticancer and cardioprotection. Remarkably, optimal derivative 43 displayed promising bi-functional effects, especially in various self-established heart failure mice models with and without tumor-bearing. Collectively, the present study validated the practicability of the "trilogy of drug repurposing" strategy in the development of bi-functional co-therapy agents.

A benzenesulfonic acid-modified organic polymer monolithic column with reversed-phase/hydrophilic bifunctional selectivity for capillary electrochromatography / 药物分析学报

Here,a styrene-based polymer monolithic column poly(VBS-co-TAT-co-AHM)with reversed-phase/hydrophilic interaction liquid chromatography(RPLC/HILIC)bifunctional separation mode was success-fully prepared for capillary electrochromatography by the in situ polymerization of sodium p-styrene sulfonate(VBS)with cross-linkers 3-(acryloyloxy)-2-hydroxypropyl methacrylate(AHM)and 1,3,5-triacryloylhexahydro-1,3,5-triazine(TAT).The preparation conditions of the monolith were optimized.The morphology and formation of the poly(VBS-co-TAT-co-AHM)monolith were confirmed by scanning electron microscopy(SEM)and Fourier transform infrared spectroscopy(FT-IR).The separation perfor-mances of the monolith were evaluated systematically.It should be noted that the incorporation of VBS functional monomer can provide π-π interactions,hydrophilic interactions,and ion-exchange in-teractions.Hence,the prepared poly(VBS-co-TAT-co-AHM)monolith can achieve efficient separation of thiourea compounds,benzene series,phenol compounds,aniline compounds and sulfonamides in RPLC or HILIC separation mode.The largest theoretical plate number for N,N'-dimethylthiourea reached 1.7×105 plates/m.In addition,the poly(VBS-co-TAT-co-AHM)monolithic column showed excellent reproducibility and stability.This novel monolithic column has great application value and potential in capillary electrochromatography(CEC).

Construction of multi-enzyme cascade reactions and its application in the synthesis of bifunctional chemicals / 生物工程学报

The synthesis of fine chemicals using multi-enzyme cascade reactions is a recent hot research topic in the field of biocatalysis. The traditional chemical synthesis methods were replaced by constructing in vitro multi-enzyme cascades, then the green synthesis of a variety of bifunctional chemicals can be achieved. This article summarizes the construction strategies of different types of multi-enzyme cascade reactions and their characteristics. In addition, the general methods for recruiting enzymes used in cascade reactions, as well as the regeneration of coenzyme such as NAD(P)H or ATP and their application in multi-enzyme cascade reactions are summarized. Finally, we illustrate the application of multi-enzyme cascades in the synthesis of six bifunctional chemicals, including ω-amino fatty acids, alkyl lactams, α, ω-dicarboxylic acids, α, ω-diamines, α, ω-diols, and ω-amino alcohols.

Expression and characterization of a bifunctional thermal β-glucosidase IuBgl3 from thermophilic archaeon Infirmifilum uzonense / 生物工程学报

β-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring β-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family β-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 ℃, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 ℃ for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl β-d-glucoside (pNPG) and p-nitrophenyl β-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters Km and Vmax were 0.38 mmol and 248.55 μmol/(mg·min), respectively, and the catalytic efficiency kcat/Km was 6 149.20 s-1mmol-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the β-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.

D-bifunctional protein deficiency caused by / 中国当代儿科杂志

A 15-day-old boy was admitted to the hospital due to repeated convulsions for 14 days. The main clinical manifestations were uncontrolled seizures, hypoergia, feeding difficulties, limb hypotonia, and bilateral hearing impairment. Clinical neurophysiology showed reduced brainstem auditory evoked potential on both sides and burst-suppression pattern on electroencephalogram. Measurement of very-long-chain fatty acids in serum showed that C26:0 was significantly increased. Genetic testing showed a pathogenic compound heterozygous mutation, c.101C>T(p.Ala34Val) and c.1448_1460del(p.Ala483Aspfs*37), in the

Structure, function and application of serine carboxypeptidase-like acyltransferases from plants / 生物工程学报

Plant serine carboxypeptidase-like acyltransferases (SCPL-AT) have similar structural characteristics and high homology compared to the serine carboxypeptidase. They can transfer the acyl from acyl glucose esters to many natural products, participate in the acylation modification of plant secondary metabolites, enrich the structural diversity of natural products, and improve the physicochemical properties such as water solubility and stability of compounds. This review summarizes the structural characteristics, catalytic mechanism, functional characterization, and biocatalytic applications of SCPL-AT from plants. This will help to promote the functional characterization of these acyltransferase genes and the biosynthesis of useful plant secondary metabolites by synthetic biotechnology.

Role of D-bifunctional protein in promoting the growth of hepatocellular carcinoma in rats and nude mice / 临床肝胆病杂志

ObjectiveTo investigate the expression of D-bifunctional protein (DBP) in hepatocellular carcinoma (HCC) tissue in rats and the growth of DBP-induced HCC in nude mice. MethodsA total of 22 male Sprague-Dawley rats were randomly divided into normal control group with 8 rats and model group with 14 rats treated with intraperitoneally injected diethylnitrosamine to induce HCC, and Western blotting, immunohistochemistry, and RT-PCR were used to measure the expression of DBP. A total of 14 specific pathogen-free male BALB/c-nu mice were randomly divided into two groups. HepG2 cells were transfected with empty plasmid or DBP overexpression plasmid and were then injected subcutaneously into nude mice. There were 8 mice in the empty plasmid control group and 6 mice in the DBP high-expression plasmid group, and tumor size was measured for both groups. The t-test was used for comparison of continuous data between groups. ResultsThe rats with HCC had significantly higher protein and mRNA expression of DBP in liver tissue than normal rats (protein: 1.10±0.35 vs 0.67±0.12, t=-7.48, P＜0.05; mRNA: 3.70±0.85 vs 1.17±0.72, t=-20.46, P＜0.05). The DBP high-expression plasmid group had a significantly higher tumor volume than the empty plasmid group ［(7590.50±1867.97)mm3 vs (1663.78±420.24)mm3, t=-39.78, P＜0.01］. ConclusionHighly expressed DBP can promote the progression of HCC in rats and thus provides a new target for the treatment of HCC and the research and development of inhibitory drugs.

Genomic analysis of bifunctional Class C-Class D ß-lactamases in environmental bacteria

β-lactamases, which are found in several bacterial species and environments, are the main cause of resistance to β-lactams in Gram-negative bacteria. In 2009, a protein (LRA-13) with two β-lactamase domains (one class C domain and one class D domain) was experimentally characterised, and an extended action spectrum against β-lactams consistent with two functional domains was found. Here, we present the results of searches in the non-redundant NCBI protein database that revealed the existence of a group of homologous bifunctional β-lactamases in the genomes of environmental bacteria. These findings suggest that bifunctional β-lactamases are widespread in nature; these findings also raise concern that bifunctional β-lactamases may be transferred to bacteria of clinical importance through lateral gene transfer mechanisms.

A Systematic Comparative Evaluation of ⁶⁸Ga-Labeled RGD Peptides Conjugated with Different Chelators / 대한핵의학회잡지

PURPOSE: The present paper reports a systematic study on the effect of bifunctional chelators (BFC) namely, NOTA, DOTA, and DTPA, on the radiochemical formulation, in vitro stability, and in vivo biological properties of ⁶⁸Ga-labeled RGD peptide derivatives.METHODS: The three RGD conjugates namely, NOTA-Bn-E-[c(RGDfk)]₂, DOTA-Bn-E-[c(RGDfk)]₂, and DTPA-Bn-E-[c(RGDfk)]₂ were radiolabeled with ⁶⁸Ga and the radiolabeling was optimized with respect to the ligand amount, radiolabeling time, and temperature. Further, the ⁶⁸Ga complexes were assessed for their in vitro and in vivo stabilities. The biodistribution studies of the three radiolabeled conjugates were carried out in C57BL/6 mice bearing melanoma tumor at 30 min and 1 h post-adimistration.RESULTS: NOTA-Bn-E-[c(RGDfk)]₂ could be radiolabeled with ⁶⁸Ga at room temperature while DOTA-Bn-E-[c(RGDfk)]₂ and DTPA-Bn-E-[c(RGDfk)]₂ were radiolabeled at high temperature. ⁶⁸Ga-NOTA-Bn-E-[c(RGDfk)]₂ was found to be the most kinetically rigid in in vitro stability assay. The uptake of the three radiolabeled peptide conjugates in melanoma tumor was comparable at 1 h post-administration (NOTA; DOTA; DTPA (% I.D./g):: 2.78 ± 0.38; 3.08 ± 1.1; 3.36 ± 0.49). However, the tumor/background ratio of ⁶⁸Ga-NOTA-Bn-E-[c(RGDfk)]₂ was the best amongst the three radiotracers. ⁶⁸Ga-complexes of NOTA-Bn-E-[c(RGDfk)]₂ and DOTABn-E-[c(RGDfk)]₂ showed excellent in vivo stability while ⁶⁸Ga-DTPA-Bn-E-[c(RGDfk)]₂ showed significant metabolic degradation.CONCLUSION: These studies show that ⁶⁸Ga-NOTA-Bn-E-[c(RGDfk)]₂ would be the most appropriate ⁶⁸Ga-labeled radiotracer and the most amenable for kit formulation.

A brief analysis of bifunctional molecules / 药学学报

Selectivity of drug action is a determinant for wide therapeutic window and less adverse response. From the viewpoint of molecular structure the conception and strategy of drug design are mainly embodied in raising selectivity. For the target-based drug discovery it is crucial to precisely obliterate detrimental targets in dimension of time and space, so as to efficaciously translate the in vitro active compounds into in vivo therapeutic medicines. To realize this translation drug molecules must be accurately transported to and destroy the harmful targets. To this end, chemical structures of drugs must be manipulated in multiple dimensions. This article attempts to concisely describe several kinds of bifunctional molecules for raising selectivity from the standpoint of medicinal chemistry. The bifunctionality of antibody-drug conjugates (ADCs) involves in the guidance and carrier of the antibody to guide ADC and reach to target cells, and simultaneously injury quality of the toxin moiety of ADC interacts with and destroys targets. Based upon target 3D structures design of irreversible inhibitors consist in connecting an appropriate electrophilic moiety to a well-defined ligand to endow the molecule with an additional ability to covalently bond to a specific amino acid residue. Hydrophobic tag (HyT), proteosis-targeting chimera (PROTAC), and degradation tag (dTAG) are new developed technologies, which are structurally characterized by bifunctionality, and mechanistically these compounds are capable of recruiting protein of interest (POI), inducing protein-protein interaction (PPI), and cleaving POI. In spite of large molecular size and the bottleneck of pharmacokinetic and physicochemical properties these technologies still have broad development prospect owing to high selectivity and wide adaptations.

Synthesis of Bifunctional Graphene Quantum Dots and Its Application in Fluorescence Detection of pH and Cell Imaging / 分析化学

Functional groups may change the electrical characteristics of graphene quantum dots,thus leading to the improvement of their properties and related applications. To improve the optical properties,we designed and synthesized pentaethylene hexamine and dodecylamine functionalized graphene quantum dots (PEHA-GQD-DA). Citric acid was mixed with pentaethylene hexamine and heated at 170℃ for 0.5 h. Then dodecylamine was added and the reaction was continued at 160℃ for 1.5 h to obtain PEHA-GQD-DA. The Yesults revealed that the PEHA-GQD-DA was composed of the graphene sheets from 1 nm to 3 nm,and their edges contained abundant functional groups. The introduction of pentaethylene hexamine greatly improved the fluorescence emission. The fluorescence quantum yield reached 72.7%,which was much higher than that of the GQD prepared by the thermal hydrolysis of single citric acid. The introduction of dodecylamine created a special amphiphilic structure that allows the quantum dots more likely to enter cell through the phospholipid bilayer of cell membrane. The PEHA-GQD-DA exhibited an excellent optical behavior for pH value of the medium. Within pH range of 1.0-6.5, the fluorescence intensity increased with the increase of pH value. The fluorescence spectrum sensitively changed with the change of pH value. There was a good linear relationship between the maximum emission wavelength and the pH value. When the pH was in the range of 6.5-12.0,the fluorescence spectrum didn't change with the change of pH value. However, its fluorescence intensity linearly decreased with the increase of pH value. The existence of common inorganic ions and organic small molecules does not interfere with pH response of the quantum dots. The PEHA-GQD-DA has been successfully applied to fluorescent detection of pH value in water samples and Hela cell imaging.

Efficient Water Splitting Catalyzed by Cobalt Phosphide Nanobead-chain Like Nanoarrays Supported on Three Dimensional Nickel Foam / 分析化学

The research for efficient and low-cost electrocatalysts for water splitting are essential for the exploitation and application of hydrogen energy. Transitional metal phosphides are considered as one of the most promising bifunctional water splitting electrocatalysts. Herein, we reported a facile two-step method (firstly hydrothermal preparation, then phosphorization under low temperature) to synthesize the bead-chain like nanoarrays of CoP supported on three dimensional Nickel foam (CoP/NF). The as-prepared CoP/NF could act as an efficient bifunctional catalyst for overall water splitting. The catalyst exhibited remarkable activity for both the oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) in alkaline media,delivering a current density of 10 mA/cm2at an overpotential of 281 mV for OER and 95 mV for HER, respectively. Efficient water splitting in alkaline system was realized by applying a two-electrode system with the CoP/NF acting as both the anode and cathode. The applied potential was 1.63 V to obtain the current density of 10 mA/cm2,and good stability was also testified.

A novel bifunctional xylanase/cellulase TcXyn10A from Thermoascus crustaceus JCM12803 / 生物工程学报

Efficient utilization of cellulose and xylan is of importance in the bioethanol industry. In this study, a novel bifunctional xylanase/cellulase gene, Tcxyn10a, was cloned from Thermoascus crustaceus JCM12803, and the gene product was successfully overexpressed in Pichia pastoris GS115. The recombinant protein was then purified and characterized. The pH and temperature optima of TcXyn10A were determined to be 5.0 and 65-70 °C, respectively. The enzyme retained stable under acid to alkaline conditions (pH 3.0-11.0) or after 1-h treatment at 60 °C. The specific activities of TcXyn10A towards beechwood xylan, wheat arabinoxylan, sodium carboxymethyl cellulose and lichenan were (1 480±26) U/mg, (2 055±28) U/mg, (7.4±0.2) U/mg and (10.9±0.4) U/mg, respectively. Homologous modeling and molecular docking analyses indicated that the bifunctional TcXyn10A has a single catalytic domain, in which the substrate xylan and cellulose shared the same binding cleft. This study provides a valuable material for the study of structure and function relationship of bifunctional enzymes.

Role of nociceptin/orphanin FQ opioid peptide receptor in pain modulation / 国际药学研究杂志

Pain is not only a common syndrome in clinic,but also a disease harming people′s health and quality of life. Dis?covery of potent and low-or non-addictive analgesic agent is a great challenge and our expectation. Nociceptin/orphanin FQ opioid pep?tide (NOP)receptor,the fourth member of the opioid receptor family,was discovered in 1994. Growing evidence has revealed that NOP receptor plays an important role in pain transduction and modulation and becomes a potential target for novel analgesics develop?ment. This review focuses on the progresses in exploring the biological characteristics of NOP receptor and its complex role in pain modulation,as well as the discovery of novel analgesic agents targeting NOP receptor,which provides reference for understanding the mechanisms of pain and analgesia and finding ideal analgesics.

Role of nociceptin/orphanin FQ opioid peptide receptor in pain modulation / 国际药学研究杂志

Pain is not only a common syndrome in clinic, but also a disease harming people’s health and quality of life. Discovery of potent and low-or non-addictive analgesic agent is a great challenge and our expectation. Nociceptin/orphanin FQ opioid peptide (NOP)receptor, the fourth member of the opioid receptor family, was discovered in 1994. Growing evidence has revealed that NOP receptor plays an important role in pain transduction and modulation and becomes a potential target for novel analgesics development. This review focuses on the progresses in exploring the biological characteristics of NOP receptor and its complex role in pain modulation, as well as the discovery of novel analgesic agents targeting NOP receptor, which provides reference for understanding the mechanisms of pain and analgesia and finding ideal analgesics.

Bifunctional Fluorescence Molecular Probes for Detection of Aluminum and pH / 分析化学

A fluorescent molecular probe R6G-Flu was prepared by modifying fluorescein onto Rhodamine 6G.The probe could be used to recognize Al3+ specifically, and the detection limit could reach as low as 10-8 mol/L.After addition of Al3+ (10 μmol/L) to the probe, the solution showed a color change from colorless to pink, and green fluorescence was observed under the UV irradiation, which could be perceived by the naked eye.By measuring the fluorescence emission intensity of R6G-Flu at different pH, the probe could also be used to determine pH in acidic pH range (3.00-6.00) and basic pH range (8.00-10.50).The detection results of Al3+ and pH indicated that the R6G-Flu was a dual-functional fluorescent molecular probe.

Modification of mPEGylated novel bifunctional fusion protein GAD and its propenty / 中国药科大学学报

In order to solve the difficucties of renaturation and immunogenicity of new bifunctional fusion protein GAD,inclusion bodies of GAD were modified by PEG-maleimide.Conformational changes of the modified GAD were compared by circular dichroism and tryptophan fluorescence spectroscopy.The biological activity was verified by oral glucose tolerance test and lipid scavenging.The results showed that PEG-maleimide completed the specific-point modification of GAD,and improved its refolding efficiency.The secondary and tertiary structures of mPEGylated GAD were consistent with that of GAD.PEG-GAD has significant hypoglycemic and lipid-lowering effects (P ＜0.001) with longer half life in vivo and lower immunogenicity (P ＜0.01).This study provides effective strategies for the development of strongly hydrophobic peptide drugs.

Progress of research and development of antibody-based therapeutics / 中国药学杂志

OBJECTIVE: With the rapid progress and continuous development of modern biotechnology, the antibody-based biotherapeutics have emerged as a leading force and play an important role in the field of biotechnology industry.

Impact of D-bifunctional Protein Deficiency on Telomere Length and Gene Expression in a Child.

Aim: To explore, in one patient, the possibility that D-bifunctional protein (D-BP) deficiency affects telomere length, and to determine the profile of genetic expression. Presentation of Case: Due to the symptoms of a newborn and his family background, a peroxisomal panel was performed. There were high levels of very long chain fatty acids and abnormal peroxisomes. At 8 months the patient exhibited other complications, including progressive multi systemic deterioration, and at 15 months died of pneumonia. Discussion: Analysis of the patient’s fibroblasts provided evidence of a defect in the peroxisomes and in the oxidation of fatty acids, leading to a diagnosis of D-BP deficiency. Significant alterations were found in the genetic expression profile, with the greatest number of affected genes involved in neuronal functions, two implicated in peroxisomal biogenesis, and some others related to telomere protection and DNA repair. The child had a mixture of very short and normal length telomeres, a condition commonly observed in the elderly and in individuals with chronic degenerative diseases. Conclusion: The abnormal function of peroxisomes and altered gene expression found in the patient under study could explain the affected telomere length. Further studies are needed to explore this possibility.

Radiolabeling of NOTA and DOTA with Positron Emitting 68Ga and Investigation of In Vitro Properties / 핵의학분자영상

PURPOSE: We established radiolabeling conditions of NOTA and DOTA with a generator-produced PET radionuclide 68Ga and studied in vitro characteristics such as stability, serum protein binding, octanol/water distribution, and interference with other metal ions. MATERIALS AND METHODS: Various concentrations of NOTA.3HCl and DOTA.4HCl were labeled with 1 mL 68GaCl3 (0.18~5.75 mCi in 0.1 M HCl) in various pH. NOTA.3HCl (0.373 mM) was labeled with 68GaCl3 (0.183~0.232 mCi/0.1 M HCl 1.0 mL) in the presense of CuCl2, FeCl2, InCl3, FeCl3, GaCl3, MgCl2 or CaCl2 (0~6.07 mM) at room temperature. The labeling efficiencies of 68Ga-NOTA and 68Ga-DOTA were checked by ITLC-SG using acetone or saline as mobile phase. Stabilities, protein bindings, and octanol distribution coefficients of the labeled compounds also were investigated. RESULTS: 68Ga-NOTA and 68Ga-DOTA were labeled optimally at pH 6.5 and pH 3.5, respectively, and the chelates were stable for 4 hr either in the reaction mixture at room temperature or in the human serum at 37 degreesC. NOTA was labeled at room temperature while DOTA required heating for labeling. 68Ga-NOTA labeling efficiency was reduced by CuCl2, FeCl2, InCl2, FeCl3 or GaCl3, however, was not influenced by MgCl2 or CaCl2. The protein binding was low (2.04~3.32%). Log P value of 68Ga-NOTA was -3.07 indicating high hydrophilicity. CONCLUSION: We found that NOTA is a better bifunctional chelating agent than DOTA for 68Ga labeling. Although, 68Ga-NOTA labeling is interfered by various metal ions, it shows high stability and low serum protein binding.