Antiproliferative Properties of Vinyl Dipeptides: Synthesis and MCF-7 Cell Line Testing

Peptide mimics derived with close structure to peptide have vast utility because they are expected to interfere with biological targets while having superior drug-like properties if compared to peptides. In this work, novel vinyl dipeptides which are different in a double bond between the α-carbon of peptide and C1 of its side chain. Added to that, suitable substituents were selected to harness drug-like properties. The compounds were found to have moderate activities when tested against MCF-7 breast cancer cell line. For instance, the adamantyl analogue 2-(benzoylamino)-3-(2-furyl)-N-(1-adamantyl) propenamide (1c) and the heterocyclic analogue 2-(Benzoylamino)-3-(2-furyl)-N-[2-(5-cyanothia-zol-2-yl)] propenamide (1o) exhibited inhibition potency at 27.4 and 37.8 μM, respectively.

Systematic screening and structural characterization of dipeptides using offline 2D LC-LTQ-Orbitrap MS:A case study of Cordyceps sinensis

Cordyceps sinensis(C.sinensis)is a widely used and highly valuable traditional Chinese medicine.Several dipeptides have been detected in C.sinensis,but current scientific knowledge of its chemical makeup remains limited.In this study,an improved approach that integrates offline two-dimensional liquid chromatography(2D LC)separation,precursor ion list,library screening,and diagnostic ion filtering was established to systematically screen and characterize dipeptides in C.sinensis.Offline 2D LC integrating hydrophilic interaction LC and reverse phase separations was established to eliminate interference and identify the target dipeptides.A library containing the potential 400 dipeptides was created,and a precursor ion list with all theoretical precursor ions was adopted to trigger the MS/MS scan with high sensitivity.To identify dipeptides,the type and connection sequence of amino acids were determined according to the product ions.Ile and Leu residues were differentiated for the first time according to the characteristic ion at m/z 69.07.Ultimately,170 dipeptides were identified or tentatively characterized from C.sinensis,and most are reported for the first time in this species herein.In addition,the identified dipeptides were also applied for discrimination among the three Cordyceps species,and 11 markers were identified.The obtained results provide a deeper understanding of the chemical basis of C.sinensis.

DETERM INATION OF THE STABILITY CONSTANTS OF COMPLEXES OF HISTIDINE,SOME DIPEPTIDES AND TRIGLYCINE WITH Zn(Ⅱ)AND Cd(Ⅱ)USING SMALL-SCALE PH-METRIC TITRATIONS

A potentiometric study on the complexes of His,Gly-His,ALa-His,Gly-Gly and Gly—Gly—Gly with Zn(Ⅱ)and Cd(Ⅱ)has been reported.Small-scale potentiometric titrations were car- ried out to determine stabil ity constants of complexes at 25℃ with I=0.10 mol dm^(-3)(KNO_3).The com- puter programs SUPERQUAD were applied for data treatment with satisfactory results.

Proline-based Amino Pyridine Dipeptides as Efficient Organocatalysts for Direct Aldol Reaction

A series of proline-based amino pyridine dipeptide organocatalysts was synthesized and applied in direct asymmetric intermolecular aldol reaction. These catalysts showed good solubility in organic solvents, good yields （73%--97%） and enantioselectivitives（74%--94%）. Among them, dipeptide organocatalyst（2） was found to be the most efficient one for the asymmetric aldol reaction between cyclohexanone and 4-nitrobeznaldehyde. After optimizing the catalytic reaction conditions, we found that the catalyst showed high yield（97%）, enantioselectivity（e.e., up to 92%） and anti-diastcreoselectivity（up to 95：5） at mild room temperature without any additives.

A New Metal Tag for Highly Selective and Sensitive Analyses of Amino Acids and Dipeptides by HPLC/ICP-MS

We have developed a novel metal tag, bis(ethylenediamine)-4'-methyl-4-carboxybipyridine-ruthenium N-succinimidyl ester (ECRS) for sensitive analysis of amino acids using high performance liquid chromatography/inductively coupled plasma mass spectrometry (HPLC/ICP-MS). ECRS is a functional reagent, containing an ester group at one end that can be activated to bind to amino group and a chelated ruthenium at the other. The activated ester was reacted briefly with amino groups under weakly alkaline conditions. The ruthenium was detected sensitively by ICP-MS. ECRS was reacted with 17 proteinogenic amino acids in borate buffer. The derivatives were separated by reversed phase HPLC and identified by quadrupole-based ICP-MS. ECRS was suitable for speciation;low molecular weight compounds containing amino groups. We have thus established a quantitative analytical method for amino acids and dipeptides. The detection limits of branched amino acids (signal-to-noise ratio of 3) were 1.5 nmol·L-1 in the standard solution (100 amol per injection).

Inhibition of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase enzyme by dipeptides identified in dry-cured ham

High cholesterolemia is a key risk factor for the development of cardiovascular diseases,which are the main cause of mortality in developed countries.Most therapies are focused on the modulation of its biosynthesis through 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMG-CoAR)inhibitors.In this sense,food-derived bioactive peptides might act as promising health alternatives through their ability to interact with crucial enzymes involved in metabolic pathways,avoiding the adverse effects of synthetic drugs.Dry-cured ham has been widely described as an important source of naturally-generated bioactive peptides exerting ACEI-inhibitory activity,antioxidant activity,and anti-inflammatory activity between others.Based on these findings,the aim of this work was to assess,for the first time,the in vitro inhibitory activity of HMG-CoAR exerted by dipeptides generated during the manufacturing of dry-cured ham,previously described with relevant roles on other bioactivities.The in vitro inhibitory activity of the dipeptides was assessed by measuring the substrate consumption rate of the 3-hydroxy-3-methylglutaryl CoA reductase in their presence,with the following pertinent calculations.Further research was carried out to estimate the possible interactions of the most bioactive dipeptides with the enzyme by performing in silico analysis consisting of molecular docking approaches.Main findings showed DA,DD,EE,ES,and LL dipeptides as main HMG-CoAR inhibitors.Additionally,computational analysis indicated statin-like interactions of the dipeptides with HMG-CoAR.This study reveals,for the first time,the hypocholesterolemic potential of dry-cured ham-derived dipeptides and,at the same time,converges in the same vein as many reports that experimentally argue the cardiovascular benefits of dry-cured ham consumption due to its bioactive peptide content.

Influence of peptide bond on photosensitized oxidation of tryptophan, tyrosine and histidine dipeptides

IN photodynamic therapy, the damage of protein results from the photooxidation of one ormore of only five amino acid residues (tryptophanyl, tyrosyl, histidyl, methionyl and cys-teinyl). These five amino acids are also readily photooxidized in their monomeric states. How-ever, the reaction kinetics and mechanisms of free amino acids and bound amino acid residuesin proteins are somewhat different. For example, Tassin found that the incorporation of tryp-tophan into a peptide linkage dramatically altered its susceptibility to photooxidation as well

Theoretical Study on the Cyclization Mechanism of Dipeptides

Cyclization is an important chemical reaction for the dipeptides containing N-alkyl groups. The cyclization mechanism has been examined by theoretical calculations. Our calculation results indicate that the most favorable mechanism is the piperidine-catalyzed stepwise mechanism, in which piperidine acts as a proton shuttle. The attack of the N-terminal amino nitrogen at the C-terminal carbonyl carbon along with the proton transfer is the rate-limiting step. The effect of the alkyl substituent on the amide N on the eyclization reaction was then examined. Finally, the influence of the solvation effect, electronic effect and steric effect on the cyclization was investigated. It is found that all of these effects contribute to the cyclization.

Dock-able linear and homodetic di, tri, tetra and pentapeptide library from canonical amino acids: SARS-CoV-2 Mpro as a case study

Peptide-based therapeutics are increasingly pushing to the forefront of biomedicine with their promise of high specificity and low toxicity.Although noncanonical residues can always be used,employing only the natural 20 residues restricts the chemical space to a finite dimension allowing for comprehensive in silico screening.Towards this goal,the dataset comprising all possible di-,tri-,and tetra-peptide combinations of the canonical residues has been previously reported.However,with increasing computational power,the comprehensive set of pentapeptides is now also feasible for screening as the comprehensive set of cyclic peptides comprising four or five residues.Here,we provide both the complete and prefiltered libraries of all di-,tri-,tetra-,and penta-peptide sequences from 20 canonical amino acids and their homodetic(N-to-C-terminal)cyclic homologues.The FASTA,simplified molecular-input line-entry system(SMILES),and structure-data file(SDF)-three dimension(3D)libraries can be readily used for screening against protein targets.We also provide a simple method and tool for conducting identity-based filtering.Access to this dataset will accelerate small peptide screening workflows and encourage their use in drug discovery campaigns.As a case study,the developed library was screened against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)main protease to identify potential small peptide inhibitors.

Peptide-based enteral formula improves tolerance and clinical outcomes in abdominal surgery patients relative to a whole protein enteral formula

AIM To compare a dipeptide- and tripeptide-based enteral formula with a standard enteral formula for tolerance and nutritional outcomes in abdominal surgery patients.METHODS A retrospective study was performed to assess the differences between a whole-protein formula(WPF) and a dipeptide- and tripeptide-based formula(PEF) in clinical outcomes.Seventy-two adult intensive care unit(ICU) patients with serum albumin concentrations less than 3.0 g/d L were enrolled in this study.Patients were divided into two groups(WPF group = 40 patients,PEF group = 32 patients).The study patients were fed for at least 7 d,with ≥ 1000 m L of enteral formula infused on at least 3 of the days.RESULTS The mean serum albumin level on postoperative day(POD) 10,prealbumin levels on POD-5 and POD-10,and total lymphocyte count on POD-5 were significantly higher in the PEF group compared to those in the WPF group(P < 0.05).The average maximum gastric residual volume of the PEF patients during their ICU stays was significantly lower than that for WPF patients.CONCLUSION Dipeptide- and tripeptide-based enteral formulas are more efficacious and better tolerated than wholeprotein formulas.

In-silico Prediction of the Sweetness of Aspartame Analogues from QSPR Analysis

The extensive utilization of the low-energy dipeptide sweetener aspartame in foods leads to various studies on searching for new sweeteners in series. However, the real mechanistic cause of their sweetness power is still not completely known owing to their complex interactions with human sweet receptor, which may be different from that of other sweeteners to some extent. In this contribution, predictive quantitative structure-property relationship(QSPR) models have been developed for diverse aspartame analogues using Materials Studio 5.0 software. The optimal QSPR model(r2 = 0.913, r2 CV = 0.881 and r2 pred = 0.730) constructed by the genetic function approximation method has been validated by the tests of cross validation, randomization, external prediction and other statistical criteria, which shows that their sweetness power is mainly governed by their electrotopological-state indices(SssCH and SsNH), spatial descriptors(Shadow length: LX, ellipsoidal volume and Connolly surface occupied volume) and topological descriptors(Chi(3): cluster and Chi(0)(valence modified)), which partially supports both multipoint attachment theory proposed by Nofre and Tinti et al. and B-X theory proposed by Kier et al.. Present exploited results provide the key structural features for the sweetness power of aspartame analogues, supplement the mechanistic understanding of the sweet perception, and would be also helpful for the design of potent sweetener analogs prior to their synthesis.

In silico identification and characterization of common epitope-based peptide vaccine for Nipah and Hendra viruses

Objective: To explore a common B-and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus(He V) and Nipah virus(Ni V). Methods: Membrane proteins F, G and M of He V and Ni V were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B-and T-cell epitopes that shared maximum identity with He V and Ni V were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico. Results: One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope(VDPLRVQWRNNSVIS) showed at least 66% identity with all Ni V and He V G protein sequences, while the 15-mer M-epitope(GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all Ni V and He V M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II(MHC II) and class-I(MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response. Conclusions: Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against He V and Ni V. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.

Carnosine concentration and expression profiles of carnosine related genes in Mytilus after beta-alanine injection

Carnosine and its analogues are histidine-containing dipeptides(HCDs)playing diverse functions in vertebrates.However,the distribution and the metabolism of carnosine in invertebrates are still unknown.In this study,Mytilus coruscus,a shellfish with important economic value in China,was selected for the investigation of HCD content and the expression profiling of carnosine-related genes in various mussel tissues.Quantification of HCD by amino acids analyzer revealed a low concentration of anserine in muscular tissues in Mytilus,indicating the presence of HCD even in an invertebrate.mRNA expression of five carnosine metabolic-related genes was profiled in various tissues,and the results highlighted the relative higher expression level of these genes in muscular tissues.Considering the fact that beta-alanine supplementation can increase the HCD content in vertebrates,a beta-alanine injection was performed and the changes of HCD concentration and the mRNA expression of carnosine related genes were investigated in five mussel tissues.The results revealed the increase of HCD concentration,as well as the up-regulated expression level of related genes,in tested tissues of beta-alanine injected mussels.Transcriptomic analysis for the whole soft tissue of mussel before and after beta-alanine injection were performed,and 3569 differential expression genes(DEGs)were identified in the beta-alanine injected group when compared to their expression levels in the control.These data indicated the complex eff ects of betaalanine on M.coruscus metabolism,and those DEGs enriched in pathways of cancers,muscle contraction,and tyrosine metabolism highlighted the possible functions of beta-alanine in cell proliferation,sports,and melanogenesis,respectively.

Novel Efficient Method for Synthesis of N_(1,4)-Disubstituted-1,4-benzodiazepine-2,5-diones

A novel and efficient method was developed for the liquid-phase synthesis of Nj,4-disubstitutedbenzodiazepine-2,5-diones with 2-chloro-5-nitrobenzoic acid as initiating material via 4 step reactions containing esterifieation, ＂Ulmn reaction＂, acylation, alkylation and cyclization. The reaction conditions were mild and the overall yields of the products ranged from 45% to 71%.

First Results of ACH Cardioplegic Solution Clinical Application in Newborns and Infants under One Year of Age

Study objective involved comparison of two cardioplegic solutions: HTK-solution possessing high buffer capacity and a new ACH-solution with aminoacid buffer. Results revealed high cardioprotective efficiency during surgical repair of complex congenital heart disease both in the group that had received Custodiol and in the group that had received ACH-solution. Clinical and morphological parameters demonstrate high level of myocardial protection from intraoperative ischemia for single usage of ACH-solution during cardioplegic ischemia under 60 minutes in duration.

Simultaneous determination of doxorubicin and its dipeptide prodrug in mice plasma by HPLC with fluorescence detection

A simple and sensitive high performance liquid chromatography with fluorescence detection （HPLC-FD） has been developed for simultaneous quantification of doxorubicin （DOX） and its dipeptide conjugate prodrug （PDOX） in mice plasma. The chromatographic separation was carried out on an Amethyst C18-H column with gradient mobile phase of 0.1% formic acid and 0.1% formic acid in acetonitrile at a flow rate of 1.0 mL/min. The excitation and emission wavelengths were set at 490 and 550 nm, respectively. The method was comprehensively validated. The limits of detection were low up to 5.0 ng/mL for DOX and 25.0 ng/mL for PDOX. And the limits of quantification were low up to 12.5 ng/mL for DOX and 50 ng/mL for PDOX, which were lower than those for most of the current methods. The calibration curves showed good linearity （R2 〉 0.999） over the concentration ranges. The extraction recoveries ranged from 84.0% to 88.2% for DOX and from 85.4% to 89.2% for PDOX. Satisfactory intra-day and inter-day precisions were achieved with RSDs less than 9.1%. The results show that the developed HPLC-FD method is accurate, reliable and will be helpful for preclinical pharmacokinetic study of DOX and PDOX.

The Effect of Glycyl-Glutamine Dipeptide Concentration on Enzyme Activity, Cell Proliferation and Apoptosis of Jejunal Tissues from Weaned Piglets

An experiment was conducted in a singly factorial design to study the effect of glycyl-glutamine dipeptide on enzyme activity, cell proliferation and apoptosis of jejunal tissues from weaned piglets at different glycyl-glutamine concentration levels of 2, 4, 10, 20, and 30 mmol L-1, respectively. The glutaminase activity, diamine oxidase （DAO） activity, cell peoliferation, apoptosis, and perotein metabolism were measured by the tissue culture method in vitro using jejunal tissues. The immunohistochemical method was used to study the cell proliferation and apoptosis of jejunal tissues. The results showed that compared to the blank control, the percentage and MOD value of BrdU-positicve cells incubated with glycyl-glutamine dipeptide solution were significantly （P0.05） increased. Accordingly, the percentage and MOD value of caspase-3-positive cells from tissue incubated with glycyl-glutamine dipeptide were notably lower （P0.05） than that from the control treatment. The glycyl-glutamine dipeptide increased the glutaminase activity, DAO activity and protein content of jejunal tissues, as the dipeptide concentration was on the rise （P0.05）. These results indicated that glycyl-glutamine dipeptide affected the jejunum development and adaptation of weaned piglets, and the function might be fulfilled by enhancing the glutamine-related enzyme activity, thereby increasing the consumption of glutamine, and then improving the jejunal cell proliferation and suppressing cell apoptosis. The effects of glycyl-glutamine dipeptide relied in a dose-dependent manner, and the maximum effect was achieved at 20-30 mmol L-1 glycyl-glutamine dipeptide.

Science Letters:EHPred: an SVM-based method for epoxide hydrolases recognition and classification

A two-layer method based on support vector machines (SVMs) has been developed to distinguish epoxide hydrolases (EHs) from other enzymes and to classify its subfamilies using its primary protein sequences. SVM classifiers were built using three different feature vectors extracted from the primary sequence of EHs: the amino acid composition (AAC), the dipeptide composition (DPC), and the pseudo-amino acid composition (PAAC). Validated by 5-fold cross tests, the first layer SVM clas- sifier can differentiate EHs and non-EHs with an accuracy of 94.2% and has a Matthew’s correlation coefficient (MCC) of 0.84. Using 2-fold cross validation, PAAC-based second layer SVM can further classify EH subfamilies with an overall accuracy of 90.7% and MCC of 0.87 as compared to AAC (80.0%) and DPC (84.9%). A program called EHPred has also been developed to assist readers to recognize EHs and to classify their subfamilies using primary protein sequences with greater accuracy.

Synthesis of Muramyl dipeptide Analogs by Incorporation of 3,3,3-Trifluoroalanine

Carbobenzoxy-L-3, 3, 3-trifluoroalanine was synthesized and it was incorporated into MDP for replacement of L-alanine.