javid shahbazi mojarrad

Methotrexate (MTX) is one of the most effective therapeutics to treat different types of solid tumors; however, it suffers low permeability limiting its bioavailability and cellular uptake. To tackle this, we aim to design and fabricate different types of cell-penetrating peptides (CPPs) to improve the intracellular uptake of MTX without causing any immunogenic response. CPPs were synthesized by the solid-phase peptide synthesis method. Peptide-MTX conjugates were prepared via covalent binding of peptide and drug molecule. CPPs and peptide-E8 nanoparticles were characterized using zeta-sizer and scanning electron microscopy. Cytotoxicity of CPPs and peptide-MTX conjugates was evaluated by MTT assay. An enzyme-linked immunosorbent assay was employed to assess the IL-6 and TNF-α cytokine release profile. Amongst all sequences, W4R4-MTX possessed the highest loading efficiency (97%) and drug to peptide percentage (24.02%). The lowest loading efficiency (36%) and drug to peptide percentage (8.76%) were seen for NGRWK-MTX conjugates. The NGRWR peptide and NGRWR-E8 nanoparticles had acceptable size (~100 nm) with spherical and rod-like structures, respectively. The selected CPPs and peptide-MTX conjugates did not show any cytotoxicity or immunogenicity. The fabricated peptides are represented as promising carriers to improve the intracellular delivery of MTX to cancer cells with low immunogenic and cytotoxic effects on normal cells.

Doxorubicin (Dox) is one of the most well-known chemotherapeutics that are commonly applied for a wide range of cancer treatments. However, in most cases, efflux pumps like P-glycoprotein (P-gp), expel the taken drugs out of the cell and decrease the Dox bioavailability. Expression of P-gp is associated with elevated mRNA expression of the ATP-binding cassette B1 (ABCB1) gene.

In the current study, different sequences of cell-penetrating peptides (CPPs) containing tryptophan, lysine, and arginine and their nano-complexes were synthesized and their impact on the expression and activity of the ABCB1 gene was evaluated in the A549 lung carcinoma cell line. Furthermore, the cellular uptake of designed CPPs in the A549 cell line was assessed.

The designed peptides, including [W4K4], [WR]3-QGR, R10, and K10 increased Dox cytotoxicity after 48 hr. Furthermore, arginine-rich peptides showed higher cellular uptake. Rhodamin123 accumulation studies illustrated that all the obtained peptides could successfully inhibit the P-gp pump. The designed peptides inhibited the ABCB1 gene expression, of which, [W4K4] resulted in the lowest expression ratio.

[W4K4], [WR]3-QGR, R10, and K10 could successfully increase the Dox cytotoxicity by decreasing the efflux pump gene expression.

The emergence of multidrug-resistant (MDR) Pseudomonas aeruginosa andAcinetobacter baumanniiis a profound therapeutic concern requiring innovative antimicrobials. Nanotechnology has unlocked new promises, and nanoparticles (NPs) have been used either alone or in combinations against MDR bacteria to overcome antibacterial resistance.

We aimed to evaluate the antibacterial activity of silver and chitosan nanoparticles alone and in combination with either ciprofloxacin or gentamicin against A. baumannii and P. aeruginosa strains.

The antibacterial activities of synthesized materials were evaluated using microdilution broth, disc diffusion, and drop diffusion methods and the expression of target genes by the quantitative real-time polymerase chain reaction.

The result of disc diffusion and drop diffusion methods for chitosan when assessed against MDR or wild type P. aeruginosa and MDR A. baumannii at different concentrations ranged from 8 - 35 mm and 6 - 15 mm respectively, while the range was lower in the nanoparticle form of chitosan. The results also showed that the expression of efflux pump adeB in A. baumannii and mexB in P. aeruginosa decreased after treatment with chitosan, chitosan nanoparticles, and silver nanoparticles; thus, they may be good candidates as efflux pump inhibitors.

The present study showed the superiority of antibacterial and anti-efflux pump activity of chitosan over silver nanoparticles against MDR P. aeruginosa and A. baumannii

In this study, a series of piperazin-2-one derivatives were prepared through bioisosteric substitution of the imidazole ring of L-778,123 (imidazole-containing FTase inhibitor) and rearrangement of groups based on the tipifarnib structure. Final compounds were evaluated for their cytotoxic activities on cancer and normal cell lines by MTT assay.

Methyl α-bromophenylacetic acid and 1-(3-chlorophenyl) piperazin-2-one were synthesized using previously described methods. Methyl 2-(4-chlorophenyl)-2-(4-(3- chlorophenyl)-3-oxopiperazin-1-yl) acetate was prepared by reaction between these two compounds in presence of potassium carbonate. Finally, methoxy group of ester was substituted by various amines such as guanidine, thiourea, urea and hydrazide. The synthesized compounds were tested for their cytotoxicity against colon cancer (HT-29) and lung cancer (A549) cell lines as well as MRC-5 (normal fetal lung fibroblasts) cells as a healthy cell line using MTT colorimetric assay method.

Replacement of imidazole moiety with guanidine, thiourea, and hydrazide could increase cytotoxicity toward all three cell lines. Some substituents, such as amine, urea, and hydroxylamine exhibited significant cytotoxicity (<500 µM) but lower than L-778,123 as standard compound. Hydroxyl and methoxy substituents did not show significant cytotoxicity. Imidazole substituent group revealed cytotoxicity similar to L-778,123 All compounds showed lower cytotoxic activity against normal cell lines compared with cancer cell lines.

It seems the electron density of substituted groups and rearrangement of groups may significantly increase cytotoxic activity

Many antimicrobial medications are available to combat infections. However, the indiscriminate use of antibiotics has produced antibiotic resistance in the case of many bacterial pathogens. This study focuses on the development of nanoparticles (NPs) that enhance the in vitro antibiotic activity of vancomycin against multi-drug resistant (MDR) organisms.
Spherical shaped thioglycolic acid-stabilized silver nanoparticles (TGA-AgNPs) were prepared by using a simple chemical reduction method. Then, vancomycin was conjugated to the terminal carboxyl of TGA in the presence of N-Hydroxysuccinimide (NHS) and N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC). Afterwards, the antibacterial activity of these nanoconjugates was examined by using the minimum inhibitory concentration (MIC) assay against MDR bacteria.
The rate of vancomycin bound to the AgNPs was 19.6%. The MIC values of vancomycin (Van)-capped AgNPs against tested pathogens were in the range of (3.2, 1.6, 0.8, 0.4, 0.2, 0.1, 0.05, and 0.025 µl/ml). The MIC was 0.1 µg/ml for VRE, MIC≤0.02 µg/ml for MRSE, and 0.05 µg/ml for S. aureus. The MIC corresponded to the MBC for all bacterial species.
This study indicated that some antimicrobial agents like vancomycin can be conjugated with AgNPs. This can lead to increased antimicrobial activity against MDR microorganisms.

Due to the high number of women affected by cervical cancer and the importance of an early diagnosis, combined with the frequent incidence of false‑negative Papanicolaou (Pap) smear screening results for this disease, several studies have been conducted in recent years in order to find better tests. Liquid‑based cytology (LBC) tests, including the liquid‑based thin layer method, have demonstrated the highest potential for reducing false‑negative cases and improved sample quality. This study aimed to compare the strength of the Pap smear test with fluid cytology and conventional tests in detecting cervical dysplasia.

This descriptive‑analytic study was conducted on 366 women who attended private laboratories for a Pap smear. The Pap smear sampling was conducted simultaneously using two

conventional Pap (CP) smear and LBC), from the cervix.

The mean age of the participants was 32 ± 8.8 years. Diagnostic results of endocervical cells, epithelial cells, vaginitis cells, and metaplastia were consistent with both conventional and liquid cytology smears, and the kappa coefficient was determined to be significant (P < 0.001). In total, 40.5% of diagnostic cases indicated bacterial inflammation 80.3% of the diagnoses in both methods were P1 and 3.9% of cases diagnosed were P2, the overall diagnostic consistency was 83.9% between the two sampling methods. The inflammation diagnosis was 40.5% and this was consistent in both methods of LBC and CP. There was one case of a false‑negative diagnosis in the LBC method and 14 cases in the CP method.

Results showed that the LBC may improve the sample’s quality and reduce the number of unsatisfactory cases more than with the CP method.

Cell-penetrating peptides (CPPs) are used for delivering drugs and other macromolecular cargo into living cells. In this paper, we investigated the relationship between the structural/physicochemical properties of four new synthetic peptides containing arginine-tryptophan in terms of their cell membrane penetration efficiency. The peptides were prepared using solid phase synthesis procedure using FMOC protected amino acids. Fluorescence-activated cell sorting and fluorescence imaging were used to evaluate uptake efficiency. Prediction of the peptide secondary structure and estimation of physicochemical properties was performed using the GOR V method and MPEx 3.2 software (Wimley-White scale, helical wheel projection and total hydrophobic moment). Our data showed that the uptake efficiency of peptides with two tryptophans at the C- and N-terminus were significantly higher (about 4-fold) than that of peptides containing three tryptophans at both ends. The distribution of arginine at both ends also increased the uptake efficiency 2.52- and 7.18-fold, compared with arginine distribution at the middle of peptides. According to the obtained results the value of transfer free energies of peptides from the aqueous phase to membrane bilayer could be a good predictor for the cellular uptake efficiency of CPPs.

Breast cancer is the second leading cancer type among people of advanced countries. Various methods have been used for cancer treatment such as chemotherapy and radiotherapy. In the present study we have designed and synthesized a new group of drug delivery systems (DDS) containing a new class of Cell Penetrating Peptides (CPPs) named Peptide Amphiphiles (PAs). Two PAs and anionic peptides were synthesized using solid phase peptide synthesis (SPPS), namely [KW]4, [KW]5, E4 and E8. Then nano-peptides were synthesized by non-covalent binding between PAs and poly anions as [KW]4-E4, [KW]4- 8, [KW]5-E4 and [KW]5-E8. Flow cytometry studies showed that increased chain length of PAs with a higher ratio between hydrophobicity and net charge results in increased intracellular uptake by MCF7 cells after 2h incubation. Moreover, nano-peptides showed greater intracellular uptake compared to PAs. Anti-proliferative assay revealed that by increasing chain length of PAs, the toxicity effect on MCF7 cells is reduced, however nano-peptides did not show significant toxicity on MCF7 cells even at high concentration levels. These data suggest that due to the lack of toxicity effect at high concentration levels and also high cellular uptake, nano-peptides are more suitable carrier compared to PAs for drug delivery.

Interference with microtubule polymerization results in cell cycle arrest leading to cell death. Colchicine is a well-known microtubule polymerization inhibitor which does so by binding to a specific site on tubulin. A set of 3',4'-bis (substituted phenyl)-4'H-spiro[indene-2,5'-isoxazol]-1(3H)-one derivatives with known antiproliferative activities were evaluated for their tubulin binding modes. 3D structures of the derivatives were docked into the colchicine binding site of tubulin using GOLD 5.0 program under flexible ligand and semi-flexible receptor condition. The spiroisoxazoline derivatives bind tubulin in a similar manner to colchicine by establishing at least a hydrogen bonding to Cys241 as well as hydrophobic interactions with Leu255, Ile378 and Lys254 and few other residues at the binding pocket. It can be concluded that the spiroisoxazoline core structure common to the studied derivatives is a suitable scaffold for placing the antitubulin pharmacophoric groups in appropriate spatial positions required for tubulin binding activity.

Farnesyltransferase (FTase) is a zinc-dependent enzyme that adds a farnesyl group to the Ras proteins. L778, 123 is a potent peptidomimetic imidazole-containing FTase inhibitor. L778123 was synthesized according to known methods and evaluated alone and in combination with doxorubicin against A549 (adenocarcinomic human alveolar basal epithelial cells) and HT29 (human colonic adenocarcinoma) cell lines by MTT assay. L778123 showed weak cytotoxic activity with IC50 of 100 and 125 for A549 and HT-29 cell lines, respectively. The combination of doxorubicin and L778123 can decrease IC50 of doxorubicin in both cell lines significantly. It can be concluded that L778, 123 can be a good agent for combination therapy.

The renin angiotensin system which is stimulated by angiotensin II, leads to increase in blood pressure. An angiotensin II receptor antagonist can control effectively hypertension. Synthesis of new compounds that have the key structural elements present in angiotensin receptor antagonists is of interest. Herein, we report the synthesis of novel Alkyl 2-azido-3-[1-[[2''-(tetrazol-5-yl) biphenyl-4-yl]methyl]-2-alkylimidazolyl]acrylate (4 and 6). 2-Alkyl-4(5)-hydroxymethylimidazoles were synthesized via Weidenhagen reaction using copper salt, an aldehyde, dihydroxyacetone and concentrated ammonium hydroxide. 2-Alkyl-4(5)-hydroxymethylimidazoles were oxidized by heating with MnO2 in CH2Cl2:dioxane to its carboxaldehyde derivatives. The carboxaldehyde derivatives were reacted with tritylated [4´-(bromomethyl) biphenyl-2-yl]tetrazole in presence of potassium carbonate to give two regioisomers 1 & 2 that separated by column chromatography. The regioisomers 1 & 2 were employed in the synthesis of 2-azidoacrylate derivatives 3 & 5 using sodium alkoxide and alkyl α-azidoacetate. Consequently, the protecting group of tetrazole was removed by stirring the compound 3 and 5 in 10% hydrochloric acid solution at room temperature. The regioisomers 1 & 2 were separated chromatographically in 1:3 proportions using 70: 30 toluene: ethyl acetate as eluent, respectively. The alkyl 3-substituted imidazol-2-azido acrylate derivatives were synthetized as oily form in 19-25% yield. The deprotection of tetrazole was afforded light yellow oily final compounds in 60-65% yield. Four new alkyl α-azidoacrylate derivatives bearing biphenyl tetrazole moiety were synthetized via Knoevenagel condensation. The protecting group was removed in acidic media to afford final compounds 4 and 6. The chemical structures of synthesized compounds were characterized by FT-IR and 1HNMR spectroscopies

DHPEE is a newly synthesized compound by merging the key structural elements in an angiotensin receptor blocker (Telmisartan) with key structural elements in 1,4- dihydropyridine calcium channel blocker (Nifedipine). In this study, we examined dual calcium channel blocking and AT1 antagonist activity for DHPEE. The functional inhibitory characteristics of DHPEE were studied in vitro in rat thoracic aorta preparations precontracted by phenylephrine (1µM) or KCl (80µM) or Ang II in normal or calcium-free solutions. Concentration–dependent significant relaxation was observed in aortic rings precontracted with phenylephrine, KCl or Ang II. The tension increment produced by increasing external calcium was also reduced by DHPEE. DHPEE caused a marked decrease in the maximal contractile response of the vasoactive agents and shifted their concentration-response curves to the right. DHPEE possesses dual characteristics and cause vasorelaxation by blocking the L-type calcium channels and blocking Ang II receptors (AT1) in rat aortic smooth muscle.

We report the synthesis of novel 1,4-dihydropyridine derivatives containing biphenyl-2''-tetrazole moieties. We hypothesized that merging the key structural elements present in an AT1 receptor antagonist with key structural elements in 1,4-dihydropyridine calcium channel blockers would yield novel analogs with potential dual activity for both receptors. This strategy led to the design and synthesis of dialkyl 1,4-dihydro-2,6-dimethyl-4-[2-n-alkyl-1-[2΄-(1H-tetrazole-5-yl) biphenyl -4-yl] methyl] imidazole-4(or 5)-yl]- 3, 5-pyridinedicarboxylate analogs. These compounds were obtained by two methods starting from biphenyltetrazolyl-4-(or 5)-imidazolecarboxaldehyde intermediates employing in classical Hantzsch condensation reaction. In the first method, triphenylmethyl protecting group of 4- or 5-carboxaldehyde intermediate was first removed in acidic media and then classical Hantzsch reaction was employed in order to obtain the final products. In the second method, without further deprotection process, protected 4- or 5-carboxaldehyde intermediate directly was used in Hantzsch reaction. The second method was more efficient than the first method since the deprotection and ring closure reaction occurs simultaneously in one pot. Eight novel dihydropridines analogs were synthesized using classic Hantzsch condensation reaction. Chemical structures of the compounds were characterized by 1H NMR, infrared and mass spectroscopy.