Hemp products are currently enjoying much commercial interest as potential anti-inflammatory preparations. However, evidence for efficacy is lacking for some clinical conditions and for crude extracts and mixtures of cannabinoids. Uterine inflammation (endometritis) is a major cause of subfertility in livestock. The aim was to assess whether a commercially important crude hemp extract (extracted with supercritical CO2) and a commercial CBG:CBD (cannabigerol:cannabidiol) cannabinoid mixture have anti-inflammatory properties, using an ex vivo bovine tissue culture model of endometritis. LC-MS2 (liquid chromatography-tandem mass spectrometry) analysis of the crude extract allowed identification of twelve cannabinoids (cannabidiolic acid (CBDA), cannabidiol (CBD), cannabidivarinic acid (CBDVA), cannabidivarin (CBDV), cannabielsoic acid A (CBEA-A), cannabielsoic acid B (CBEA-B), cannabichromene (CBC), cannabinoloic acid (CBNA), cannabinol (CBN), cannabiripsol (CBR), cannabicoumarononic acid (CBCONA), cannabicoumaronone (CBCON)) in the extract, along with isomers, an alkaloid and three polyunsaturated fatty acids. Fragmentation pathways, some including 6-jr-electron pericyclic processes, are proposed. Tissue explants, from uteri from heifers at stage I or IV of the oestrous cycle, were treated with control, LPS (lipopolysaccharide), crude hemp extract, or with LPS + crude hemp extract for 24 h. The CBG:CBD mixture was assessed similarly. Inflammatory responses were measured as concentrations of PGF2a, PGE2 and IL-6 in culture supernatants. Secretion of IL6 from tissue explants was abrogated by the hemp extract (50mg mL-1), compared to LPS alone, but it had no effect on secretion of PGF2a. The CBG:CBD mixture had no effect on any of the markers. The crude hemp extract was not cytotoxic towards MDBK bovine kidney cells whereas the CBG:CBD mixture (50 mg mL-1) caused cell death. The hemp extract warrants further investigation for anti-inflammatory effects using a broader range of inflammatory markers, whereas the CBG:CBD mixture is not recommended for further study in the context of bovine endometritis. (c) 2025 The Authors. Published by Elsevier B.V. on behalf of SAAB. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

Urease catalyses the hydrolysis of urea to ammonia and carbon dioxide. This enzyme is important in the virulence of several human pathogens and urease activity in soil can cleave urea fertilisers prematurely, leading to waste of agricultural nitrogen. A series of arylsulfonylurea-glucosamine hybrid compounds were synthesised. Reaction of arylsulfonamides with phenyl chloroformate and 4-dimethylaminopyridine gave either phenyl N-(2,4-arylsulfonyl)carbamate 4-dimethylaminopyridinium salts or N-(4-arylsulfonyl)-4-dimethylaminopyridinium-1-carboxamide inner salts, depending on the substitution on the arylsulfonamide. Both types of intermediates, gave ester-protected arylsulfonylurea-glucosamines, when treated with 1,3,4,6-tetra-O-acetylglucosamine. Simple methanolysis gave the arylsulfonylurea-glucosamine hybrids as interconverting mixtures of anomers. Both the O-acetyl intermediates and the target arylsulfonylurea-glucosamines inhibited jack-bean urease with IC 50 10–36 μM. This narrow range of values precluded the determination of structure-activity relationships and docking studies suggested several different optimum docking poses for the various analogues. No analogues showed radical-scavenging activity. Several compounds showed modest cytotoxic activity against renal carcinoma cells in the NCI 60-cell-line screen.

Aromatase inhibitors are among the most effective treatment of the breast cancer. Aromatase catalyzes estrogen biosynthesis, which is a long-term cause of breast cancer. Current study describes the synthesis, purification of 26 new fluorinated and non-fluorinated thiourea derivatives of desloratadine (5), and their aromatase inhibition activity, cytotoxicity against cancer cell line (MDA-MB-231). Compounds 7 v and 7 l exhibited a significant anti-aromatase activity, while compounds 7 a, 7 g-h, 7 m and 7 u were also significant active against MDA-MB-231 cell line. Furthermore, the molecular docking studies revealed that active compounds form key interactions with the crucial amino acid of aromatase active site including TRP224, LEU477, CYS437, ALA438, MET374, ARG115, ILE305, and PHE221, which are responsible for the binding interactions of aromatase. All analogues were new, except 7 b and 7 k and also lacked cytotoxicity against BJ human fibroblasts, with the exception of 5 and 7 x. This selectivity makes this series particularly interesting for further studies.

Aim: We aim to develop new anti-leishmanial agents against Leishmania major and Leishmania tropica.Materials & methods: A total of 23 thiourea derivatives of (±)-aminoglutethimide were synthesized and evaluated for in vitro activity against promastigotes of L. major and L. tropica.Results & conclusion: The N-benzoyl analogue 7p was found potent (IC50 = 12.7 μM) against L. major and non toxic to normal cells. The docking studies, indicates that these inhibitors may target folate and glycolytic pathways of the parasite. The N-hexyl compound 7v was found strongly active against both species, and lacked cytotoxicity against normal cells, whereas compound 7r, with a 3,5-bis-(tri-fluoro-methyl)phenyl unit, was active against Leishmania, but was cytotoxic in nature. Compound 7v was thus identified as a hit for further studies.

An understanding of basic organic chemical reactivity is key for many biosciences students. The reactivities of different carbonyl groups affect their function in many biomolecules. A practical class, the two-step synthesis of paracetamol, has been devised to illustrate the electrophilic reactivities of carbonyls, which was covered in the accompanying lecture program. Students also examine the UV, IR, NMR, and mass spectra of the esters and amides, building further on the understanding gained in lectures. The practical work itself has been devised to be able to be run in bioscience laboratories with limited chemical facilities. The class has been enhanced during four academic years with strong support from the students.

Clutia lanceolata is a medicinal plant native to Ethiopia and sub-Saharan Africa and to the Arabian Peninsula. It is used traditionally in Saudi Arabia for the treatment of diabetes. Previous phytochemical analysis of this species has been limited to the identification of methylthiocoumarins. Further work has led to isolation of 19 new diterpenoids in three structural classes. Their structures were established by HRMS and by a range of NMR techniques (1H, 13C, COSY, NOESY, HSQC, HMBC), with confirmation for some examples by X-ray crystallography. NOESY and 1H-1H NMR coupling constants gave the relative stereochemical configurations and conformational information, with absolute configurations being established through X-ray crystallography. One example closely related to the known hypoglycemic compound saudin (found in C. richardiana and also in C. lanceolata) and one with a different core tetracycle were found to enhance strongly the glucose-triggered release of insulin from murine pancreatic islets. Biosynthetic proposals for the three groups of new diterpenoids by alternative cyclization of a common precursor are put forward. Lanceolide P (16) is proposed as a lead compound for further development for the treatment of diabetes.

Rare actinomycetes are a source of numerous diverse, biologically active secondary metabolites, including macrolides, which have been shown to display several antibiotic activities. The bioactivities and representative structures of 26 groups of macrolides from rare actinomycetes are presented in this review. The most interesting groups, with a wide range of biological activities, are ammocidins, bafilomycins, neomaclafungins, rosaramicins, spinosyns, and tiacumicins. Most macrolides are from the genus, Micromonospora, with smaller contributions from genera such as Saccharothrix, Amycolatopsis, Nocardiopsis and Catenulispora. These macrolides display unique cytotoxic, antibacterial, antifungal, antimicrobial, insecticidal, anti-trypanosomal, antimalarial, antiprotozoal, antimycobacterial and anti-herpetic activity. Considering their bioactivities and diverse structures, macrolides from rare actinomycetes warrant further investigation for future applications in medicine. This work highlights the bioactivities and structures of important classes of macrolides from rare actinomycetes that are already marketed or could be used in medicine in the future.

Racemases and epimerases catalyse changes in the stereochemical configurations of chiral centres and are of interest as model enzymes and as biotechnological tools. They also occupy pivotal positions within metabolic pathways and, hence, many of them are important drug targets. This review summarises the catalytic mechanisms of PLP-dependent, enolase family and cofactor-independent racemases and epimerases operating by a deprotonation/reprotonation (1,1-proton transfer) mechanism and methods for measuring their catalytic activity. Strategies for inhibiting these enzymes are reviewed, as are specific examples of inhibitors. Rational design of inhibitors based on substrates has been extensively explored but there is considerable scope for development of transition-state mimics and covalent inhibitors and for the identification of inhibitors by high-throughput, fragment and virtual screening approaches. The increasing availability of enzyme structures obtained using X-ray crystallography will facilitate development of inhibitors by rational design and fragment screening, whilst protein models will facilitate development of transition-state mimics.

Teucrium yemense , a medicinal plant commonly grown in Saudi Arabia and Yemen, is traditionally used to treat infections, kidney diseases, rheumatism, and diabetes. Extraction of the dried aerial parts of the plant with methanol, followed by further extraction with butanol and chromatography, gave twenty novel neoclerodanes. Their structures, relative configurations and some conformations were determined by MS and 1-D and 2-D NMR techniques. Most were fairly conventional but one contained an unusual stable orthoester, one had its (C-16)-(C-13)-(C-14)-(C-15) (tetrahydro)furan unit present as a succinic anhydride and one had a rearranged carbon skeleton resulting from ring-contraction to give a central octahydroindene bicyclic core, rather than the usual decalin. Mechanisms are proposed for the biosynthetic formation of the orthoester and for the ring-contraction. Four novel neoclerodanes increased the glucose-triggered release of insulin from isolated murine pancreatic islets by more than the standard drug tolbutamide, showing that they are potential leads for the development of new anti-diabetic drugs.

PARP10 is an intracellular mono-ADP ribosyltransferase and recent reports suggest that it regulates proliferation of some cell types. However, its effect on the proliferation of colorectal carcinoma cells has not yet been systematically reported. We explored the influence of PARP10 on the proliferation of several colorectal carcinoma cell types and carried out initial studies on the underlying mechanisms. Inhibition of the enzymatic activity of PARP10 led to significantly decreases in proliferative ability in LoVo cells and CT26 cells in vitro and suppressed growth of CT26 tumours in the subaxilliary region in Balb/c mice in vivo. Cell-cycle arrest accompanied these observations. Expression of the nuclear transfer factor β-catenin and it trans-location to the nucleus were also affected and the expression of its associated signal proteins Axin2 and c-Myb were increased and decreased, respectively. We demonstrate that PARP10 promotes proliferation of those colorectal carcinoma cells which express significant levels of PARP10. This promotion is suppressed when the enzymatic activity is inhibited. β-Catenin is likely to be the mediator of the antiproliferative effect.

PARP10 is an intracellular mono-ADP ribosyltransferase and recent reports suggest that it regulates proliferation of some cell types. However, its effect on the proliferation of colorectal carcinoma cells has not yet been systematically reported. We explored the influence of PARP10 on the proliferation of several colorectal carcinoma cell types and carried out initial studies on the underlying mechanisms. Inhibition of the enzymatic activity of PARP10 led to significantly decreases in proliferative ability in LoVo cells and CT26 cells in vitro and suppressed growth of CT26 tumours in the subaxilliary region in Balb/c mice in vivo. Cell-cycle arrest accompanied these observations. Expression of the nuclear transfer factor β-catenin and it translocation to the nucleus were also affected and the expression of its associated signal proteins Axin2 and c-Myb were increased and decreased, respectively. We demonstrate that PARP10 promotes proliferation of those colorectal carcinoma cells which express significant levels of PARP10. This promotion is suppressed when the enzymatic activity is inhibited. β-Catenin is likely to be the mediator of the antiproliferative effect.

Thieno[2,3-c]isoquinolin-5(4H)-one is known for its potential as an anti-ischemic agent through the inhibition of poly(ADP-ribose) polymerase 1 (PARP1). However, the compound also inhibits many other enzymes of the PARP family, potentially limiting its usability. The broad inhibition profile, on the other hand, indicates that this molecule backbone could be potentially used as a scaffold for the development of specific inhibitors for certain PARP enzymes. These efforts call for novel synthetic strategies for substituted thieno[2,3-c]isoquinolin-5(4H)-one that could provide the needed selectivity. In this article, an efficient synthetic strategy for 8-alkoxythieno[2,3-c]isoquinolin-5(4H)-ones through eight steps is presented and other tested synthetic pathways are discussed in detail. Synthesis of 7-methoxythieno[2,3-c]isoquinolin-5(4H)-one is also demonstrated to show that the strategy can be applied widely in the syntheses of substituted alkoxythieno[2,3-c]isoquinolin-5(4H)-ones.

Clutia lanceolata Forssk. (C. lanceolata), is a medicinal plant native to sub-Saharan Africa and the Arabian Peninsula. Phytochemical investigation of the aerial parts of C. lanceolata yielded twenty-one coumarins (1-21), including methylthio and methylsulfinyl-coumarins. Thirteen of these compounds (1-13) are reported here for the first time, named as cluteolin A to M. The remaining eight compounds (14-21) are known but have not been associated previously with C. lanceolata. The structures of the new compounds were elucidated from their 2D NMR and MS spectra. Single crystal X-ray analysis confirmed the structures of 1,8,9,11,12,14,17,19,20. As, in Saudi Arabian tradition, C. lanceolata has been reported to have anti-diabetic and anti-fungal properties, the coumarins were examined for their biological activity. Seven compounds (2-4,8,15-17) strongly enhanced the glucose-triggered release of insulin by murine pancreatic islets, with 4 and 16 showing more than two-fold enhancement of insulin secretion, compared with the standard drug glimepiride.

α-Methylacyl-CoA racemase (AMACR; P504S; EC 5.1.99.4) catalyses epimerization of 2-methylacyl-CoAs and is important for the degradation of branched-chain fatty acids and the pharmacological activation of ibuprofen and related drugs. It is also a novel drug target for prostate and other cancers. However, development of AMACR as a drug target has been hampered by the difficulties in assaying enzyme activity. Consequently, reported inhibitors have been rationally designed acyl-CoA esters, which are delivered as their carboxylate prodrugs. The novel colorimetric assay for AMACR based on the elimination of 2,4-dinitrophenolate was developed for high-throughput screening and 20,387 'drug-like compounds' were screened, with a throughput of 768 compounds assayed per day. Pyrazoloquinolines and pyrazolopyrimidines were identified as novel scaffolds and investigated as AMACR inhibitors. The most potent inhibitors have IC50 values of ~2 µM. The pyrazoloquinoline inhibitor 10a displayed uncompetitive inhibition, whilst 10j displayed mixed competitive inhibition. The pyrazolopyrimidine inhibitor 11k displayed uncompetitive inhibition. This is the first report of the identification of specific drug-like small-molecule AMACR inhibitors by high-throughput screening. Pyrazoloquinolines and pyrazolopyrimidines may also be useful as inhibitors of other CoA-utilizing enzymes.

alpha-Methylacyl-CoA racemase (AMACR; P504S) catalyses an essential step in the degradation of branched-chain fatty acids and the activation of ibuprofen and related drugs. AMACR has gained much attention as a drug target and biomarker, since it is found at elevated levels in prostate cancer and several other cancers. Herein, we report the synthesis of 2-(phenylthio)propanoyl-CoA derivatives which provided potent AMACR inhibitory activity (IC50 = 22-100 nM), as measured by the AMACR colorimetric activity assay. Inhibitor potency positively correlates with calculated logP, although 2-(3-benzyloxyphenylthio)propanoyl-CoA and 2-(4-(2-methylpropoxy) phenylthio)propanoyl-CoA were more potent than predicted by this parameter. Subsequently, carboxylic acid precursors were evaluated against androgen-dependent LnCaP prostate cancer cells and androgen-independent Du145 and PC3 prostate cancer cells using the MTS assay. All tested precursor acids showed inhibitory activity against LnCaP, Du145 and PC3 cells at 500 mu M, but lacked activity at 100 mu M. This is the first extensive structure-activity relationship study on the influence of side-chain interactions on the potency of novel rationally designed AMACR inhibitors.

Clutia lanceolata Forssk. (C. lanceolata) is a medicinal plant native to sub-Saharan Africa and the Arabian Peninsula. Phytochemical investigation of the aerial parts of C. lanceolata yielded twenty-one coumarins including methylthio and methylsulfinyl-coumarins. Thirteen of these compounds are reported here for the first time, named as cluteolin A to M. The remaining eight compounds are known but have not been associated previously with C. lanceolata. The structures of the undescribed compounds were elucidated from their 2D NMR and MS spectra. Single crystal X-ray analyses confirmed the structures of eleven compounds. As, in Saudi Arabian tradition, C. lanceolata has been reported to have anti-diabetic and anti-fungal properties, the coumarins were examined for their biological activity. Seven compounds strongly enhanced the glucose-triggered release of insulin by murine pancreatic islets, with two compounds showing more than two-fold enhancement of insulin secretion, compared with the standard drug glimepiride.

Retama raetam is a bush which is a member of the family Fabaceae. It is used traditionally in North Africa and Saudi Arabia for the treatment of diabetes. Several flavonoids and alkaloids are already known from this plant. Chromatographic fractionation and purification led to the isolation of three new derivatives of prenylated flavones, retamasin C-E, and four new derivatives of prenylated isoflavones, retamasin F-I, in addition to two isoflavones which have not been previously reported in this plant. Particularly interesting structures included isoflavones containing 3,5-dihydro-2H-2,5-methanobenzo[e][1,4]dioxepine and 3a,8b-dihydro-7-hydroxyfuro[3,2-b]benzo[2,1-d]furan units, both of which are new amongst natural product flavonoids. Five new examples (two flavones and three isoflavones) strongly enhanced the glucose-triggered release of insulin by murine pancreatic islets and one isoflavone was a potent inhibitor of α-glucosidase. This study may rationalise the traditional medicinal use of R. raetam and provide new leads for drug design in the treatment of diabetes.

alpha-Methylacyl-CoA racemase (AMACR; P504S) is a promising novel drug target for prostate and other cancers. Assaying enzyme activity is difficult due to the reversibility of the 'racemisation' reaction and the difficulties in the separation of epimeric products; consequently few inhibitors have been described and no structure-activity relationship study has been performed. This paper describes the first structure-activity relationship study, in which a series of 23 known and potential rational AMACR inhibitors were evaluated. AMACR was potently inhibited (IC50= 400-750 nM) by ibuprofenoyl-CoA and derivatives. Potency was positively correlated with inhibitor lipophilicity. AMACR was also inhibited by straight-chain and branched-chain acyl-CoA esters, with potency positively correlating with inhibitor lipophilicity. 2-Methyldecanoyl-CoAs were ca. 3-fold more potent inhibitors than decanoyl-CoA, demonstrating the importance of the 2-methyl group for effective inhibition. Elimination substrates and compounds with modified acyl-CoA cores were also investigated, and shown to be potent inhibitors. These results are the first to demonstrate structure-activity relationships of rational AMACR inhibitors and that potency can be predicted by acyl-CoA lipophilicity. The study also demonstrates the utility of the colorimetric assay for thorough inhibitor characterisation.

Peptide-like self-immolative molecular clips are required for release of active drugs from prodrugs by endopeptidases, generating diketopiperazines (DKPs) as by-products. Two diastereomeric series of cyclo-L-aminoacyl-R/S-dimethyloxazolidine carboxylate DKPs, derived from L-Ala, L-Leu and L Val, were synthesised. The oxazolidines were constructed by acid-catalysed condensation of N-protected L-Aaa-L/D-SerOMe dipeptides with Me2C(OMe)(2), providing the protected 2,2-dimethyloxazolidinecarboxylates (Dmo) dipeptides. Deprotection exposed the primary amines, which cyclised rapidly to give the desired DKPs. The conformations were studied by nuclear magnetic resonance in CDCl3 solution and, for the L, R series, by X-ray crystallography. The L, S series had the DKP in a boat conformation with the oxazolidine in a half-chair; the L, R series had the DKP in a flattened conformation with the oxazolidine in an alternative half-chair. Early kinetic studies have shown that L, R-Dmo dipeptide amides cyclised more rapidly than L, S-diastereoisomers and that the rate ofcyclisation in the L, R series depends inversely on steric bulk at the alpha-carbon. Cyclo-L-Leu-R-Dmo was non-toxic toward human HT29 cells. [GRAPHICS] .