Pharmacological Interest Research Articles

2-Alkyl-Substituted-4-Amino-Thieno[2,3-d]Pyrimidines: Anti-Proliferative Properties to In Vitro Breast Cancer Models.

Thienopyrimidines are structural analogs of quinazolines, and the creation of new 2-alkyl derivatives of ethyl 4-aminothienopyrimidine-6-carboxylates for the study of their anti-proliferative properties is of great pharmacological interest. Some 2-alkyl-4-amino-thieno[2,3-d]pyrimidines 2-5 were synthesized, and their cyto- and phototoxicity against BALB 3T3 cells were established by an in vitro 3T3 NRU test. The obtained results indicate that the tested compounds are not cytotoxic or phototoxic, and that they are appropriate to be studied for their anti-proliferative and anti-tumor properties. The anti-proliferative potential of the compounds was investigated on MCF-7 and MDA-MB-231 cancer cells, as well as a MCF-10A cell line (normal human mammary epithelial cells). The most toxic to MCF-7 was thienopyrimidine 3 with IC50 13.42 μg/mL (IC50 0.045 μM), followed by compound 4 (IC50 28.89 μg/mL or IC50 0.11 μM). The thienopyrimidine 4 revealed higher selectivity to MCF-7 and lower activity (IC50 367 μg/mL i.e., 1.4 μM) than compound 3 with MCF-10A cells. With respect to MDA-MB-231 cells, ester 2 manifested the highest effect with IC50 52.56 μg/mL (IC50 0.16 μM), and 2-ethyl derivative 4 revealed IC50 62.86 μg/mL (IC50 0.24 μM). It was estimated that the effect of the substances on the cell cycle progression was due to cell cycle arrest in the G2 stage for MDA-MB-231, while arrest in G1 was detected for the estrogen (ER)-positive MCF-7 cell line. The tested compound's effects on the change of the zeta potential in the tumorigenic cells utilized in this study were determined. The calculation which we performed of the physicochemical properties and pharmacokinetic parameters influencing the biological activity suggested high intestinal absorption, as well as drug-likeness.

Informing the Cannabis Conjecture: From Life's Beginnings to Mitochondria, Membranes and the Electrome-A Review.

Before the late 1980s, ideas around how the lipophilic phytocannabinoids might be working involved membranes and bioenergetics as these disciplines were "in vogue". However, as interest in genetics and pharmacology grew, interest in mitochondria (and membranes) waned. The discovery of the cognate receptor for tetrahydrocannabinol (THC) led to the classification of the endocannabinoid system (ECS) and the conjecture that phytocannabinoids might be "working" through this system. However, the how and the "why" they might be beneficial, especially for compounds like CBD, remains unclear. Given the centrality of membranes and mitochondria in complex organisms, and their evolutionary heritage from the beginnings of life, revisiting phytocannabinoid action in this light could be enlightening. For example, life can be described as a self-organising and replicating far from equilibrium dissipating system, which is defined by the movement of charge across a membrane. Hence the building evidence, at least in animals, that THC and CBD modulate mitochondrial function could be highly informative. In this paper, we offer a unique perspective to the question, why and how do compounds like CBD potentially work as medicines in so many different conditions? The answer, we suggest, is that they can modulate membrane fluidity in a number of ways and thus dissipation and engender homeostasis, particularly under stress. To understand this, we need to embrace origins of life theories, the role of mitochondria in plants and explanations of disease and ageing from an adaptive thermodynamic perspective, as well as quantum mechanics.

No prescription? No problem: A qualitative study investigating self-medication with novel psychoactive substances (NPS).

The proliferation of Novel Psychoactive Substances (NPS) presents a challenge for global drug policy. The ease of online drug purchase and the emergence of the dark web have created new avenues for the growth of NPS. Despite the global nature of this issue, limited research has examined motivations of use. These include perceived safety or convenience, an interest in novel pharmacology and self-exploration. Recent evidence has suggested individuals may be self-medicating with NPS, however this phenomenon has yet to be thoroughly explored. This study aims to investigate the occurrence of NPS self-medication, identify the specific NPS involved, and understand the motivations behind their use. Discussions surrounding self-medication using NPS were collected between October 2022 and February 2023 via a content analysis of a Reddit community. Ninety-three threads, comprising 182,490 words and 5023 comments, were collected and cleaned. A frequency analysis was conducted to identify the NPS discussed, and data was analysed systematically through the process of iterative categorization (IC). Our study revealed frequent discussions about the self-medication with several NPS, notably etizolam, clonazolam, diclazepam, flualprazolam, 2-FMA, 4F-MPH, 3-FPM and 3-MeO-PCP. Individuals were mainly self-treating ADHD, anxiety and depression. Motivations for choosing NPS included access, cost, legality and a dissatisfaction with conventional healthcare. Substances were often chosen based on a profile of "Functionality" and outcomes varied. The use of clonazolam was highlighted as particularly problematic. The current study provides insight into the phenomenon of self-medication with NPS within an internet demographic, exploring the motivations behind why individuals choose NPS for a variety of disorders. The easy access to NPS and lack of scientific data pose a significant challenge for drug policy. Future policies should focus on improving healthcare providers knowledge of NPS use, removing barriers to adult ADHD diagnosis and rebuilding trust between individuals and addiction services.

Chemical composition, antioxidant and antimicrobial activities of essential oil extracted from Myrceugenia euosma (O. Berg) D. Legrand (Myrtaceae)

Essential oils (EOs) have gained notoriety due to their potential to suppress pathogenic microorganisms capable of resisting to antibiotics. It mainly happened because studies have found bioactive compounds with antimicrobial and antioxidant properties that can be used in several pharmacological and industrial applications. Brazil has a wide variety of plant species belonging to family Myrtaceae; however, studies about the pharmacological potential of its bioactive compounds remain scarce. Therefore, the aim of the current study is to investigate the chemical composition, as well as the antioxidant and antibacterial activity, of EOs extracted from fresh and dry Myrceugenia euosma (O. Berg) D. Legrand leaves. EOs were extracted both from fresh and room temperature-dried leaves, which were subjected to hydrodistillation in modified Clevenger system. EOs’ chemical composition was analyzed through Gas Chromatography coupled to Mass Spectrometry. Antioxidant activity was analyzed based on the DPPH, ABTS and FRAP methodologies. Antimicrobial activity was assessed based on determining the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of bacteria of clinical and pharmacological interest to the food, agricultural and industrial sectors, such as Bacillus cereus, Enterococcus faesium, Listeria monocytogenes, Staphylococcus aureus and Salmonella Typhimurium. Fresh-leaf EO extraction yield reached 0.06%, whereas dry-leaf EO extraction yield reached 0.11%. Based on EOS’ chemical composition analysis, spathulenol (17.55%), neointermedeol (11.22%), α-terpineol (9.1%) and eucalyptol (6.08%) were the major compounds identified in EO extracted from fresh leaves. On the other hand, α-terpineol (11.54%), spathulenol (10.24%), eucalyptol (10.1%) and neointermedeol (9.11%) were identified in EO extracted from dry leaves. Overall, the analyzed EOs have shown low antioxidant activity; there was little difference between EOs extracted fresh leaves and those extracted from dry leaves. Antimicrobial activity capable of inhibiting all tested microorganisms was observed for both EO types - MIC values ranged from 0.098 mg/mL to 0.781 mg/mL. With respect to MBC, the lowest concentrations with bactericidal action were recorded for EO extracted from fresh leaves - values ranged from 0.098 mg/mL to 0.195 mg/mL. Finally, great compound diversity was observed in the chemical composition of the investigated EOs, which presented low antioxidant activity and high antimicrobial activity. To the best of our knowledge, the current study was the first to investigate M. euosma as promising source of bioactives with potential for biotechnological application.

Could cannabinoids provide a new hope for ovarian cancer patients?

It is known that gynecological cancers remain a worldwide problem and as shown by the statistics, there is a need for new gynecological cancer treatments. Cannabinoids, the pharmacologically active compounds of the Cannabis sativa plant, have been used for many centuries by individuals as a symptomatic treatment to alleviate pain, nausea, vomiting, and to help stimulate appetite. Research has revealed that cannabinoids also exert anti-cancer activity such as anti-proliferative and pro-apoptotic effects through a variety of mechanisms. There is significant value in the development of these compounds as anti-cancer therapies in clinical practice as they do not produce the typical toxic side effects that exist with conventional therapies and recent clinical trials have shown their great tolerability by patients at high doses. Cannabinoids can induce psychoactive effects that could limit their progression. Therefore, non-psychoactive cannabinoids are attracting pharmacological interest due to their inability to produce psychological effects. Recent studies have focussed on non-psychoactive cannabinoids in ovarian cancer and have revealed promising pre-clinical results that indicate that these compounds may have potential benefits in the treatment of these cancers. However, there are still unanswered questions and research gaps that need to be addressed. This review summarizes the current understanding of this topic and identifies the current gaps in knowledge that provide a useful direction for future work.

Sonochemical synthesis, docking studies and in vitro evaluation of imidazo-1,3-oxazinone derivatives as potential inhibitors of TNF-α

A series of imidazo-1,3-oxazinone derivatives was explored as new and prospective inhibitors of TNF-α. These compounds were synthesized via a direct, one-pot method involving the coupling of appropriate bromo compound with various terminal alkynes in the presence of (PPh3)4Pd, CuI and ZnCl2 in DMF under ultrasound. This sonochemical reaction proceeded via the sequential formation of CC (coupling) and then intramolecular CO bond (cyclization) in the same pot affording the imidazo-1,3-oxazinone derivatives in good yield. A thorough docking studies carried out using the TNF-α protein (PDB: 7JRA and 6 × 81) in silico primarily with the help of AutoDock and then JAMDA docking tool predicted 3j, 3l, 3m and 3o as potential hit molecules from the current series of compounds synthesized. The majority of the compounds interacted with the common interacting residues in the TNF-α trimer e.g. TYR135, TYR227, ILE231, LEU133, LEU233, LEU196 in case of 7JRA and TYR119, TYR59, LEU57, GLY121 in case of 6 × 81. When examined in vitro against TNF-α, the compound 3l, 3m and 3o (with 69–82% inhibition) emerged as the most active compounds in this series. It was observed that the compounds possessing the tricyclic fused ring generally showed better inhibition than that having the fused bicyclic ring as noted during the docking studies. From the viewpoint of SAR (Structure Activity Relationship) the effectiveness of the C-3 aryl group in case of benzoimidazo-1,3-oxazinones was found to be in the order C6H5 > C6H4Me-p > 2-pyridyl > C6H4OMe-p > C6H4Cl-p etc. In terms of IC50 values the compound 3l, 3m and 3o (IC50 ∼5.2–6.2 µM) did not show better activities than rolipram (IC50 ∼0.9 µM) but they appeared to be superior than thalidomide (IC50 ∼197.3 µM). Thus the outcome of docking studies, in vitro evaluation and in silico ADME predictions indicated further pharmacological interest of the preliminary hits identified.

Determination of binding characteristics as a measure for effective albumin using different methods

Background & aimsTransport functions of albumin are of clinical and pharmacological interest and are determined by albumin's properties like posttranslational modifications or bound ligands. Both are affected in pathological conditions and in therapeutic grade albumin solutions. The term effective albumin concentration was introduced as a measure of functionally intact albumin. Our aim was to evaluate the impact of ligands and modifications with different approaches as a measure of effective albumin. Approach & resultsWe used a spin labelled fatty acid and dansylsarcosine to characterize binding properties of albumin i) prepared from plasma of patients and healthy control donors, ii) measured directly out of plasma, iii) research grade albumin, iv) in vitro modified albumin, and v) therapeutic infusion solutions before and after removal of stabilizers.Bilirubin is the main determinant for binding function in patients' albumin. In in vitro prepared albumin bound fatty acids correlated with impaired binding. Human nonmercaptalbumin1, not human nonmercaptalbumin2, showed reduced binding properties. Binding and transport function of therapeutic albumin was severely impaired and restored by filtration. Glycation of research grade albumin had no effect on the binding of dansylsarcosine and only a minor effect on fatty acid binding. ConclusionsOur results suggest that effective albumin -in terms of binding properties- is primarily determined by bound ligands and only to a minor extent by posttranslational modifications. Characterizing albumin directly from plasma better reflects the physiological situation whereas in the case of therapeutic grade albumin stabilizers should be removed to make its binding properties accessible.

Rational design of highly stabilized and selective adrenomedullin analogs.

The peptide hormone adrenomedullin (ADM) consists of 52 amino acids with a disulfide bond and an amidated C-terminus. Due to the vasodilatory and cardioprotective effects, the agonistic activity of the peptide on the adrenomedullin 1 receptor (AM1 R) is of high pharmacological interest. However, the wild-type peptide shows low metabolic stability leading to rapid degradation in the cardiovascular system. Previous work by our group has identified proteolytic cleavage sites and demonstrated stabilization of ADM by lipidation, cyclization, and N-methylation. Nevertheless, these ADM analogs showed reduced activity and subtype selectivity toward the closely related calcitonin gene-related peptide receptor (CGRPR). Here, we report on the rational development of ADM derivatives with increased proteolytic stability and high receptor selectivity. Stabilizing motifs, including lactamization and lipidation, were evaluated regarding AM1 R and CGRPR activation. Furthermore, the central DKDK motif of the peptide was replaced by oligoethylene glycol linkers. The modified peptides were synthesized by Fmoc/t-Bu solid-phase peptide synthesis and receptor activation of AM1 R and CGRPR was measured by cAMP reporter gene assay. Peptide stability was tested in human blood plasma and porcine liver homogenate and analyzed by RP-HPLC and MALDI-ToF mass spectrometry. Combination of the favorable lactam, lipidation, ethylene glycol linker, and previously described disulfide mimetic resulted in highly stabilized analogs with a plasma half-life of more than 144 h. The compounds display excellent AM1 R activity and wild-type-like selectivity toward CGRPR. Additionally, dose-dependent vasodilatory effects of the ADM derivatives lasted for several hours in rodents. Thus, we successfully developed an ADM analog with long-term in vivo activity.

The indications and results of the use of teduglutide in patients with short bowel.

Short bowel syndrome (SBS) is a rare condition defined as a reduced residual functional small intestinal length to less than 200 cm often resulting from extensive intestinal resection, and can lead to chronic intestinal failure (CIF). Patients with SBS-CIF are unable to absorb sufficient nutrients or fluids to maintain metabolic homeostasis through oral or enteral intake and require long-term parenteral nutrition and/or fluids and electrolytes. However, complications may arise from both SBS-IF and life-sustaining intravenous support, such as intestinal failure-associated liver disease (IFALD), chronic renal failure, metabolic bone disease and catheter-related complications. An interdisciplinary approach is required to optimize intestinal adaptation and decrease complications. In the last two decades, glucagon-like peptide 2 (GLP-2) analogs have sparked pharmacological interest as a potential disease-modifying therapy for SBS-IF. Teduglutide (TED) is the first developed and marketed GLP-2 analog for SBS-IF. It is approved in the United States, Europe, and Japan for use in adults and children with SBS-IF who are intravenous supplementation dependent. This article discusses the indications, candidacy criteria and results of the use of TED in patients with SBS.

An innovative method for the extraction and HPLC analysis of bioactive policosanols from non-psychoactive Cannabis sativa L.

Policosanols (PCs) refer to a mixture of long-chain aliphatic alcohols. Sugar cane is the main industrial source of PCs, but others, including beeswax and Cannabis sativa L., are also known. In the raw material PCs are bonded to fatty acids to form long-chain esters, known as waxes.PCs are mainly used as a cholesterol-lowering product, even though their efficacy is controversial. More recently, the pharmacological interest in PCs has increased, as they have been investigated as antioxidant, anti-inflammatory and anti-proliferative agents.Given their promising biological implications, the development of efficient extraction and analytical methodologies for the determination of PCs is extremely important to identify new potential sources of these compounds and to ensure the reproducibility of biological data. Conventional techniques used for the extraction of PCs involve time-consuming approaches leading to low yields, while analytical methods for their quantification are based on gas-chromatographic (GC) techniques, which require an additional derivatization step during the sample preparation to increase their volatility.In the light of all the above, this work was aimed at the development of an innovative method for the extraction of PCs from non-psychoactive C. sativa (hemp) inflorescences, taking advantage of the microwave-assisted technology. In addition, a new analytical method based on high-performance liquid chromatography (HPLC) coupled with an evaporative light scattering detector (ELSD) was developed for the first time for both the qualitative and quantitative analysis of these compounds in the extracts. The method was validated according to ICH guidelines, and it was applied to the analysis of PCs in hemp inflorescences belonging to different varieties. The results were analyzed using Principal Component Analysis (PCA) and hierarchical clustering analysis to rapidly identify samples with the highest content of PCs, which might find an application as alternative sources of these bioactive compounds in both the pharmaceutical and nutraceutical fields.

Physiologically-based toxicokinetic model of botulinum neurotoxin biodistribution in mice and rats

Botulinum neurotoxin (BoNT) is a highly toxic protein and a Tier 1 Biodefense Select Agent and Toxin. BoNT is also a widely used therapeutic and cosmetic. Despite the toxicological and pharmacological interest, little is known about its biodistribution in the body. The objective herein was to develop a dose-dependent, species-specific physiologically-based toxicokinetic (PBTK) model of BoNT biodistribution in rodents following a single intravenous dose. The PBTK model was based on published physiologically-based pharmacokinetic (PBPK) models of therapeutic monoclonal antibody (mAb) biodistribution because the size and charge of BoNT is nearly identical to a typical IgG4 mAb and size/charge are main factors governing protein biodistribution. Physiological compartments included the circulation, lymphatics and tissues grouped by capillary pore characteristics. Host species-specific parameters included weight, plasma volume, lymph volume/flow, and tissue interstitial fluid parameters. BoNT parameters included extravasation from blood to tissues, charge, binding to internal lamella or cholinergic neuron receptors. Parameter values were obtained from the literature or estimated using an Approximate Bayesian Computation-Sequential Monte Carlo algorithm, to fit the model to published mouse BoNT low-dose, time-course plasma concentration data. Fits captured the low-dose mouse data well and parameter estimates appeared biologically plausible. The fully-parameterized model was then used to simulate mouse high-dose IV data. Model results compared well with published data. Finally, the model was re-parameterized to reflect rat physiology. Model toxicokinetics agreed well with published rat BoNT intravenous data for two different sized rats with different intravenous doses (an a priori cross-species extrapolation). These results suggested the BoNT model predicted dose-dependent biodistribution in rodents, and for rats, without any BoNT-specific data from rats. To our knowledge, this represented a first-in-kind physiologically-based model for a large protein toxin. Results are discussed in general and in the context of human simulations to support BoNT risk assessment and therapeutic research objectives.

A Cross sectional study on Students’ Attitude towards Communication Pharmacology in Medical Education

Abstract Background: Effective communication between doctor-patient is essential to provide high-quality health care. The introduction of communication skills is a promising step at an undergraduate level by National Medical Commission (NMC) that improve students’ attitude towards patient care. Objectives: The study objective was to assess the student’s attitude and their perception towards learning communication skills. Methods: A total of 122 students participated in the study who were pursuing second-year MBBS in a Medical college located in Telangana State. A Questionnaire related to attitude towards Communication Pharmacology (Communication Skills Attitude Scale), their interest in the topic of choice and reason for their interest in communication pharmacology was given. Using the Likert scale, scores were given ranging from 13 to 65 and the median values of responses were taken for positive and negative attitudes. Results: The study results showed that the median value of positive response for girls was 56 and for boys was 52 whereas for a negative response was 25 and 27 respectively. There was no significant (p>0.05) difference was observed in the tested groups. The majority of the students opted for empathy-related communication skills as their interest (30.33%) and opined that it will help to build future communication skills (31.33%). Conclusion: It was concluded that students have a more positive attitude towards communication pharmacology and girls have shown more interest in these types of skills.

Dragon's Blood Sap Microencapsulation within Whey Protein Concentrate and Zein Using Electrospraying Assisted by Pressurized Gas Technology.

Dragon's blood sap (DBS) obtained from the bark of Croton lechleri (Müll, Arg.) is a complex herbal remedy of pharmacological interest due to its high content in polyphenols, specifically proanthocyanidins. In this paper, electrospraying assisted by pressurized gas (EAPG) was first compared with freeze-drying to dry natural DBS. Secondly, EAPG was used for the first time to entrap natural DBS at room temperature into two different encapsulation matrices, i.e., whey protein concentrate (WPC) and zein (ZN), using different ratios of encapsulant material: bioactive compound, for instance 2:1 w/w and 1:1 w/w. The obtained particles were characterized in terms of morphology, total soluble polyphenolic content (TSP), antioxidant activity, and photo-oxidation stability during the 40 days of the experiment. Regarding the drying process, EAPG produced spherical particles with sizes of 11.38 ± 4.34 µm, whereas freeze-drying produced irregular particles with a broad particle size distribution. However, no significant differences were detected between DBS dried by EAPG or freeze-drying in TSP, antioxidant activity, and photo-oxidation stability, confirming that EAPG is a mild drying process suitable to dry sensitive bioactive compounds. Regarding the encapsulation process, the DBS encapsulated within the WPC produced smooth spherical microparticles, with average sizes of 11.28 ± 4.28 µm and 12.77 ± 4.54 µm for ratios 1:1 w/w and 2:1 w/w, respectively. The DBS was also encapsulated into ZN producing rough spherical microparticles, with average sizes of 6.37 ± 1.67 µm and 7.58 ± 2.54 µm for ratios 1:1 w/w and 2:1 w/w, respectively. The TSP was not affected during the encapsulation process. However, a slight reduction in antioxidant activity measured by DPPH was observed during encapsulation. An accelerated photo-oxidation test under ultraviolet light confirmed that the encapsulated DBS showed an increased oxidative stability in comparison with the non-encapsulated DBS, with the stability being enhanced for the ratio of 2:1 w/w. Among the encapsulating materials and according to the ATR-FTIR results, ZN showed increased protection against UV light. The obtained results demonstrate the potential of EAPG technology in the drying or encapsulation of sensitive natural bioactive compounds in a continuous process available at an industrial scale, which could be an alternative to freeze-drying.

6H-Pyrimido[2,1-a]isoindoles: acid–base and complexation properties and electrooxidation model of metabolic degradation

Physicochemical properties of pharmacological interest were determined for ten 6H-pyrimido[2,1-a]isoindoles. The compounds studied were found to be weak bases with a pKa ranging from 2.38 to 3.46. Furthermore, the association constants of the studied compounds with cyclodextrins were examined. The formation of complexes was observed with 1:1 stoichiometry, γ-cyclodextrin was found to be the best complexing. Finally, the electrochemical oxidation of 6H-pyrimido[2,1-a]-isoindoles in 0.1 mol dm−3 sodium perchlorate in acetonitrile was studied as a model of their possible metabolic degradation. It was found to be a one-electron process, and the values of the half-wave potentials are in the range of 1.34–1.62 V (vs. Ag/AgNO3/NaClO4). The electrooxidation products of three selected compounds were prepared by preparative electrolysis and subsequently identified by mass spectrometry. From the data obtained, it is evident that the electrochemical oxidation of the 6H-pyrimido[2,1-a]isoindoles begins with the formation of N-oxides, followed by dimerization of the molecule or, contrary to this, by oxidative cleavage of the pyrimidine ring. From a metabolic point of view, N-oxidation is the corresponding process to this pathway.Graphical abstract

Synthesis of 2,3-dihydroquinazolin-4(1H)-one derivatives as potential inhibitors of TNF-α

'The anti-inflammatory activities of the 2,3-dihydroquinazolin-4(1H)-one derivatives along with results from our in silico predictions (this study) encouraged us to explore this class of compounds as potential inhibitors of TNF-α. Among all the molecules assessed in silico the best ranked molecules were identified with the order as 3h > 3p > 3l based on the estimated total energy. The compound 3h participated in a H-bond interaction through its C = O group of the quinazoline-4(1H)-one ring with TYR151 of chain A in addition to the interactions with all the critical TYR residues via its chloro group. The interesting docking results were also obtained for compound 3l and 3p. For the access of this class of fused N-heteroarenes we adopted the β-cyclodextrin (β-CD) catalyzed approach involving the cyclocondensation of 2-aminobenzamide with aldehydes / ketones in pure water. Various aromatic, aliphatic and heteroaromatic aldehydes (including the cinnamaldehyde) along with ketones were employed for this purpose to afford the corresponding product in good to excellent yield. The recovery and recycling of the catalyst β-CD was successful. The inexpensive and eco-friendly nature along with the simple product purification as well as good product yield are the key advantages of this approach. When examined in vitro the compound 3h, 3l and 3p with the inhibition > 70% emerged as the most active compounds in this series as predicted by in silico studies. According to the SAR (Structure Activity Relationship) the effectiveness of the C-2 aryl group was found to be in the order Ar = C6H4Cl-p > 4-pyridyl > C6H4OMe-p > C6H2(OMe)3-m,p > C6H4OH-p etc. whereas an heteroaryl or alkyl group at this position was found to be less effective or ineffective. In terms of IC50 values the compound 3h, 3l and 3p appeared as superior than the known inhibitor thalidomide though they were not better than rolipram. Being identified as preliminary hits these compounds are of further pharmacological interest.

Use of the Allium cepa Model to Assess the Cytogenotoxicity of Luffariella herdmani Marine Sponge Extract.

Marine sponge extracts are known to contain potentially toxic compounds that have biological activities of possible pharmacological interest. Thus, it is vital that biological models are used for the preliminary toxicity screening of such extracts. The present study reports the use of Allium cepa, a low-cost plant-based in vivo model, to assess the cytotoxicity and genotoxicity of Luffariella herdmani marine sponge crude extract (SCE). Pre-germinated onion bulbs, exposed for 96hours to different concentrations of SCE (ranging from 0.3125 to 20μg/ml), were used to determine general cytotoxicity. Root length as well as morphological abnormalities were recorded. Genotoxicity was assessed by exposing the root tips to SCE (0.3125-20μg/ml) and the appropriate controls for 48hours, and then staining with acetocarmine. The Mitotic Index (MI), Mitotic Phase Indices (MPIs) and chromosomal aberrations were evaluated and recorded. SCE inhibited A. cepa root growth (EC50 = 10.34μg/ml) and elicited a mitodepressive effect (LC50 = 1.95μg/ml) in a dose-dependent and significant manner. In addition, macroscopic alterations as well as chromosomal aberrations were detected. Overall, our findings indicate that L. herdmani crude extract exhibits cytotoxic and genotoxic activity, suggesting that it might contain substances with anti-proliferative/anticancer potential that could be subject to further characterisation.

Fatty acids composition of actinobacteria isolated from mussels Mytilus galloprovincialis of the Black Sea Odesa Bay

Analysis of the fatty acid composition of total cellular lipids is important chemotaxonomic characteristic that is used to identify microorganisms, in particular actinobacteria, with the help of fatty acid spectra libraries. Actinobacteria are well known as producers of secondary metabolites, which are of pharmacological and commercial interest. A significant number of actinomycetes is associated with various marine benthic communities. The aim of this study was determination of the fatty acid composition and preliminary identification of actinobacteria isolated from mussels Mytilus galloprovincialis of the Black Sea Odesa Bay. Actinobacteria of 12 isolated strains were grown in a liquid medium at 28° C for 72 h. Methyl esters of fatty acids were determined on a gas chromatograph Agilent. The MIDI Sherlock microorganisms identification system was used to identify the studied strains. It was found that the fatty acids methyl esters profiles of studied actinobacteria strains were characterized by the predominance of 12-17 saturated branched-chain fatty acids with high content of 12-methyltridecanoic, 12-methyltetradecanoic, 14-methylpentadecanoic and 14-methylhexadecanoic fatty acids. All 12 actinobacteria strains isolated from mussels (Mytilus galloprovincialis) of Odesa Bay were identified as members of the Streptomyces genus. Keywords: Black Sea, fatty acid compositions, marine actinobacteria, Mytilus galloprovincialis, Streptomyces

Advances in the phytochemical screening and biological potential of propolis.

Propolis is a natural resinous product collected from different parts of plants by bees and mixed with their salivary secretions. The occurrence of more than 180 different chemotypes has flavonoids, phenolic acids, esters, and phenolic aldehydes, as well as balsamic resins, beeswax, pollen, and essential and aromatic oils, among others. Its biological potential documented throughout the world justifies the need, from time to time, to organize reviews on the subject, with the intention of gathering and informing about the update on propolis. In this review (CRD42020212971), phytochemical advances, in vitro, in vivo, and clinical biological assays of pharmacological interest are showcased. The focus of this work is to present propolis clinical safety assays, antitumor, analgesic, antioxidant, anti-inflammatory, and antimicrobial activities. This literature review highlights propolis' promising biological activity, as it also suggests that studies associating propolis with nanotechnology should be further explored for enhanced bioprocessing applications.

Autonomous Synthesis of Functional, Permanently Phosphorylated Proteins for Defining the Interactome of Monomeric 14-3-3ζ.

14-3-3 proteins are dimeric hubs that bind hundreds of phosphorylated "clients" to regulate their function. Installing stable, functional mimics of phosphorylated amino acids into proteins offers a powerful strategy to study 14-3-3 function in cellular-like environments, but a previous genetic code expansion (GCE) system to translationally install nonhydrolyzable phosphoserine (nhpSer), with the γ-oxygen replaced with CH2, site-specifically into proteins has seen limited usage. Here, we achieve a 40-fold improvement in this system by engineering into Escherichia coli a six-step biosynthetic pathway that produces nhpSer from phosphoenolpyruvate. Using this autonomous "PermaPhos" expression system, we produce three biologically relevant proteins with nhpSer and confirm that nhpSer mimics the effects of phosphoserine for activating GSK3β phosphorylation of the SARS-CoV-2 nucleocapsid protein, promoting 14-3-3/client complexation, and monomerizing 14-3-3 dimers. Then, to understand the biological function of these phosphorylated 14-3-3ζ monomers (containing nhpSer at Ser58), we isolate its interactome from HEK293T lysates and compare it with that of wild-type 14-3-3ζ. These data identify two new subsets of 14-3-3 client proteins: (i) those that selectively bind dimeric 14-3-3ζ and (ii) those that selectively bind monomeric 14-3-3ζ. We discover that monomeric-but not dimeric-14-3-3ζ interacts with cereblon, an E3 ubiquitin-ligase adaptor protein of pharmacological interest.

In-Vitro Antioxidant, Anti-Arthritis, Anti-inflammatory, Thrombolysis, Anti-bacterial, and in-Vivo Neuropharmacological Activities of Bioactive Metabolites of Solanum americanum Mill.

Aims: The purpose of this research was to examine the effects of methanol-prepared leaf extracts from the Solanum americanum Mill. plant on a variety of in vitro activities, including antioxidant, anti-Arthritis, anti-inflammatory, thrombolysis, anti-bacterial, and in-vivo neuropharmacological activities.
 Study Design: Methanolic extract of Solanum americanum leaf (MESAL) was primarily evaluated via phytochemical screening. The potential for in vitro activities, including antioxidant, anti-arthritis, anti-inflammatory, thrombolysis, and anti-bacterial activity were determined to study due to the pharmacological interest in the plant's chemical constituent. Whether there is statistical significance to the changes seen in its in vivo neuropharmacological characteristics when tested in experimental animals.
 Place and Duration of Study: This research was carried out between November 2022 to January 2023 at the Laboratory of Phytochemistry and Pharmacology in the Department of Pharmacy and Laboratory of Microbiology, Stamford University Bangladesh and Bangladesh Council of Scientific and Industrial Research, Dhaka.
 Methodology: The plant was subjected to phytochemical screening utilizing a variety of test reagents and potential antioxidant, anti-arthritis, anti-inflammatory, thrombolytic, anti-bacterial, and neuropharmacological activities of MESAL were investigated. Antioxidant, anti-arthritic, anti-inflammatory, thrombolytic, and antibacterial activities were tested at different doses utilizing the DPPH Free Radical Scavenging Assay, Protein Denaturation Assays, Clot Lysis, and the Disk Diffusion Method. Swiss albino mice were tested using open-field and hole-cross methods to measure their locomotion as part of the neuropharmacological study.
 Results: The MESAL phytochemical screening findings demonstrated that the plants' chemical compositions varied. Most antioxidant activities were found in MESA, with an IC50 of 11.73 µg/mL compared to Ascorbic acid's IC50 of 28.86 µg/mL. When compared to the standard, the percent inhibition value of MESAL's anti-arthritis activity was significantly higher. With a maximum protein denaturation value of 94.03% at a concentration of 1000µg/mL, MESAL possesses potent anti-inflammatory activity. The value of MESAL in terms of clot lysis is very significant which is 90.257%. While MESAL's antibacterial value is moderate, it is still worth considering. However, the extract was proven to be less effective than Diazepam in improving motor coordination in the Open Field and Hole Cross Tests.
 Conclusion: Several pathological conditions, including neurodegenerative diseases, may benefit in the future from the use of plant-derived pharmacological agents due to their neuropharmacological, antioxidant, anti-inflammatory, and anti-arthritic activity, which can replace the use of NSAIDs.