Synthesis of dimeric phenol derivatives and determination of in vitro antioxidant and radical scavenging activities

In this study, di(2,6-dimethylphenol) (Di-DMP), di(2,6-diisopropylphenol) (Di-DIP, dipropofol) and di(2,6-di-t-butylphenol) (Di-DTP) were synthesized by the reaction of monomeric phenol derivatives with catalytic CuCl(OH). TMEDA and Na₂S₂O₄. Their antioxidant capacity and radical scavenging activity were examined using different in vitro methodologies such as 1,1-diphenyl-2-picryl-hydrazyl (DPPH·) free radical scavenging, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, total antioxidant activity by ferric thiocyanate, total reducing power by potassium ferricyanide reduction method, superoxide anion radical scavenging, hydrogen peroxide scavenging and ferrous ions chelating activities.     

DFT study on the radical scavenging activity of β,β-dimethylacrylalkannin derivatives

The radical scavenging activity of β,β-dimethylacrylalkannin derivatives has been studied by using density functional theory. The hydrogen bond property of the studied structures was investigated using the atoms in molecules theory. It turned out that the hydrogen bond is important for good radical scavenging activity. The hydrogen atom transfer for β,β-dimethylacrylalkannin derivatives is difficult to occur compared with the zero compound phenol. However, β,β-dimethylacrylalkannin derivatives appear to be good candidates for the one-electron transfer, particularly for β,β-dimethylacrylalkannin derivatives with electron-donating groups. Their naphthoquinone planar conformation and the extended electronic delocalisation between adjacent substituent groups determine low adiabatic ionisation potential (IP_a) values. The IP_a values of β,β-dimethylacrylalkannin derivatives with –NHPh, –N(CH₃)Ph, –N(CH₂CH₂)O and –N(CH₃)₂ groups are lower than that of the parent compound β,β-dimethylacrylalkannin, suggesting that these derivatives are expected to be the promising candidates for radical scavenging activity compounds. Taking this system as an example, we present an efficient method for the investigation of radical scavenging activity from theoretical point of view.     

Chromane derivatives of small aromatic molecules: Chemoenzymatic synthesis and growth inhibitory activity on human tumor cell line LoVo WT

Aromatic substrates tyrosol (p-hydroxyphenylethanol) and 2,6-dihydroxynaphthalene (2,6-DHN) were converted into chromane derivatives by means of chemoenzymatic reactions catalyzed by the aromatic prenyltransferase of bacterial origin NovQ, using dimethylallyl bromide as allylic substrate instead of the natural isoprenyl pyrophosphate substrate. Stereoselective prenylation occurred in o-position with respect to the phenol hydroxyl in both compounds. Prenylated derivatives were readily converted into chromane products via a selective 6-endo-trig cyclization involving the oxygen atom from the phenol moiety and the double bond of the prenyl substituent, a process catalyzed by FeCl₃. These findings set up the basis of a most convenient two-step, one-pot process which allows for easy recovery of the chromane products in high yields. The chromane derivatives thus obtained were tested for cytotoxicity and pro-apoptotic activity using LoVo WT cells, a line of human colon adenocarcinoma.     

Synthesis and Antioxidant Capacity of Some Derivatives of Sesamol at the C-6 Position

Several synthetic approaches (aminomethylation, alkylation, condensation, etc.) have been used to synthesize derivatives based on the sesamol ( 1 ), natural phenol. The set of methods, including the study of antioxidant activity (AOA) by the ability to inhibit the initiated oxidation of animal lipids, radical scavenging activity, Fe²⁺-chelation ability, as well as a comparative assessment of membrane-protective activity under the conditions of H₂O₂-induced hemolysis of mice red blood cells (RBCs), was used to analyze the antioxidant potential of the synthesized compounds. The synthesized derivatives have demonstrated different activity in the listed test systems, and we have identified compounds which appear to be most promising for a detailed study of their pharmacological properties.     

Enhancement of specific nitrogenase activity inAzospirillum brasilense andKlebsiella pneumoniae,inhibition inRhizobium japonicum under air by phenol

Specific nitrogenase activity inAzospirillum brasilense ATCC 29145 in surface cultures under air is enhanced from about 50 nmol C₂H₄·mg protein^-1·h^-1 to 400 nmol C₂H₄ by the addition of 1 mM phenol. 0.5 and 2 mM phenol added increase the rate 5-fold and 4-fold. This enhancement effect is observed only between 2 and 3 days after inoculation, with only a small reduction of the growth of the cells by the phenol added. In surface cultures under 1% O₂, nitrogenase activity is slightly reduced by the addition of 1–0.01 mM phenol. Utilization of succinate is enhanced during the period of maximum enhancement of nitrogenase activity by 60% by addition of 1 mM phenol. The cells did not produce¹⁴CO₂ from [U-¹⁴C] phenol, neither in surface cultures nor in liquid cultures and less than 0.1% of the phenol was incorporated into the cells. A smaller but significant enhancement of nitrogenase activity by about 100% in surface cultures under air was found withKlebsiella pneumoniae K 11 after addition of 1 mM phenol. However, inRhizobium japonicum 61-A-101 all phenol concentrations above 0.01 mM reduced nitrogenase activity. With 1 mM phenol added activity was reduced to less than 10% with no effect on the growth in the same cultivation system. With thisRhizobium japonicum strain significant quantities of phenol (25 mol in 24 h by 2·10¹² cells) were metabolized to¹⁴CO₂, with phenol as sole carbon source. WithAzospirillum brasilense in liquid culture under 1% and 2% O₂ in the gas phase, no enhancement of nitrogenase activity by phenol was noticed.     

Influence of various concentrations of phenol and its derivatives on the activity of fish intestinal peptidases

The effects of phenol and its derivatives (4-chlorophenol, 4-nirophenol and 2,4-dinitrophenol) on the activity of peptidases in five species of fish differing in feeding modes have been studied. The power of the effects depends on the fish species and localization of the enzyme (mucosa or chyme). As a rule, in the in vitro conditions the phenols in concentrations of 0.06?0.5 μM/L considerably decreases activities of the intestinal peptidases in bream; silver bream; and, especially, in pike. In some cases, phenol and its derivatives induce an inconsiderable increase in peptidase activities. In zander and perch, in fact, no changes in the enzymatic activity under impact of phenols are observed. It is suggested that the effect of phenol and its derivatives depends to a high extent on the species specificity of peptidases: in the fish of fam. Percidae, the enzymes are relatively tolerant; in fam. Cyprinidae and Esocidae, they are sensitive to the studied toxicants.     

Influence of various phenolic compounds on phenol hydroxylase activity of a Trichosporon cutaneum strain

The phenol-degrading strain Trichosporon cutaneum R57 utilizes various aromatic and aliphatic compounds as a sole carbon and energy source. The intracellular activities of phenol hydroxylase [EC 1.14.13.7] of a Trichosporon cutaneum R57 strain grown on phenol (0.5 g/l) were measured. Different toxic phenol derivatives (cresols, nitrophenols and hydroxyphenols) were used as substrates in the reaction mixture for determination of the enzyme activity. The data obtained showed that the investigated enzyme was capable to hydroxylate all applied aromatic substrates. The measured activities of phenol hydroxylase varied significantly depending on the aromatic compounds used as substrates. The rate of phenol hydroxylase activity with phenol as a substrate (1.0 U/mg total cell protein) was accepted as 100%.     

Photosynthetic Electron Transport Inhibition by 3-Acyl-2,4,6-trihydroxybenzamide Derivatives

A series of 3-acyl-2,4,6-trihydroxybenzamides was synthesized, and the compounds’ PET inhibitory activities were examined in isolated chloroplasts. In general, the PET inhibitory activity was found to depend on the overall lipophilicity of the molecule. Low activities of the mono and dihydroxy derivatives indicated that the three hydroxyl groups on the nucleus were essential for high activity. The PET inhibition study, on chloroplasts isolated from an atrazine resistant biotype of Brassica napus and using thermoluminescence analysis, suggested that the trihydroxybenzamide derivatives would be classified as a urea type rather than a phenol type of PET inhibitor. However the trihydroxybenzamide derivatives, like the phenol type of PET inhibitor, showed a lag time before inhibition started, which was followed by constant activity. These results indicate that the binding domain for the trihy-droxybenzamide derivatives overlapped with those of both the urea type and phenol type of PET inhibitors.     

Oxidation of phenols by cells and cell-free enzymes from Candida tropicalis

A yeast strain isolated from soil by enrichment on phenol as major carbon source was identified as Candida tropicalis. Washed cell suspensions of this strain and cell-free preparations obtained from mechanically disrupted cells oxidized phenol via catechol and cis, cis-muconate. In addition to phenol and the three isomeric diphenols, a number of phenol derivatives, amongst them fluoro-, nitro- and short-chain alkyl-phenols, were oxidized by the organism. However, no significant oxygen uptake could be demonstrated in the presence of pyrogallol, phloroglucinol, the cresols, the m-and p-hydroxy-benzoates, methoxylated phenol derivatives, benzene or toluene. Cell-free preparations from the yeast strain exhibited activity of phenol hydroxylase and of catechol 1,2-oxygenase. Both enzymes appeared in the soluble cell fraction. Both exhibit broad substrate specificities. The relative specific activity of the ring-cleaving enzyme towards various substrates seems to be dependent on the phenolic inducer.     

Characterization of a high-affinity phenol hydroxylase from Comamonas testosteroni R5 by gene cloning, and expression in Pseudomonas aeruginosa PAO1c

Comamonas testosteroni strain R5 is a phenol-degrading bacterium which expresses a phenol-oxygenating activity that is characterized by low K _s (the apparent half-saturation constant in Haldane's equation) and low K _SI (the apparent inhibition constant) values. We have now cloned the gene cluster encoding a phenol hydroxylase (phcKLMNOP) and its cognate regulator (phcR) from strain R5. Transformation of Pseudomonas aeruginosa PAO1c (Phenol⁻ Catechol⁺) with pROR502, a derivative of pRO1614 containing the cloned genes, confers the ability to grow on phenol as the sole carbon source. The K _s and K _SI values for the phenol-oxygenating activity of PAO1c(pROR502) are almost identical to those of strain R5, suggesting that the phcKLMNOP genes encode the major phenol hydroxylase in strain R5. A phylogenetic analysis shows the phenol hydroxylase from strain R5 to be more closely related to toluene/benzene-2-monooxygenase (Tb2m) from Pseudomonas sp. JS150 than to the phenol hydroxylases from P. putida CF600 and BH, or to the phenol hydroxylase from Ralstonia eutropha E2. Analysis of the substrate specificity of PAO1c(pROR502) and PAO1c derivatives expressing phenol hydroxylase from P. putida BH or from R. eutropha E2 indicates that these phenol hydroxylases catalyze the oxidation not only of phenol and cresols but also of toluene and benzene. Received: 29 March 1999 / Accepted: 18 July 1999     

Phenolic Content Pattern, Polyphenol Oxidase and Lipoxygenase Activity in Relation to Albinism, Fruit Malformation and Nubbins Production in Strawberry (Fragaria x ananassa Duch)

Total phenol content, polyphenol oxidase (PPO) and lipoxygenase (LOX) activity were determined in strawberry fruits to establish a relationship between phenolic content, PPO and LOX activity with the production of abnormal fruits in 10 different strawberry cultivars. The study revealed that all strawberry cultivars produced abnormal fruits in varying proportion. Highest percentage of abnormal fruits was produced by Etna (21.8%), and the lowest by Senga Sengana (3.7%). Among cultivars, Etna produced maximum percentage of albino (25.8%), malformed (22.7%) or nubbin (button) berries (16.9%), while the lowest percentage of albino (5.2%), and malformed fruits (2.5%) were produced by Senga Sengana, and nubbin (button) (2.9%) by Camarosa. Total phenol content and PPO and LOX activity varied widely among different strawberry cultivars. The highest phenol content was recorded in Senga Sengana (14.4 mg g^?1, fresh tissue weight) and the minimum in Etna (3.1 mg g^?1 fresh tissue weight). Likewise, Etna exhibited the minimum PPO activity (0.503 ΔA₄₀₀ g^?1 min^?1) and Senga Sengana, the maximum (0.732 ΔA₄₀₀ g^?1 min^?1). Similarly, normal fruits exhibited higher PPO activity (0.753 ΔA₄₀₀ g^?1 min^?1) than abnormal fruits. The highest LOX activity was exhibited by Senga Sengana (0.462 μ katal kg^?1) and the lowest by Etna (0.338 μ katal kg^?1) Similarly, LOX activity was very high in normal fruits (0.568 μ katal kg^?1) in comparison to abnormal ones. The regression equations developed between total phenol content, PPO and LOX activity with abnormal fruits revealed that an inverse relationship existed between total phenol content, PPO and LOX activity and production of abnormal fruits in strawberry.     

Discovery of a novel class of 2-aminopyrimidines as CDK1 and CDK2 inhibitors

A series of new 2-(2-aminopyrimidin-4-yl)phenol derivatives were synthesized as potential antitumor compounds. Substitution with pyrrolidine-3,4-diol at the 4-position of phenol provided potent inhibitory activity against CDK1 and CDK2. X-ray crystal structural studies were performed to account for the effect of the substituent on both the enzymatic and cell growth inhibitory activities.     

Computer calculation-based quantitative structure-activity relationships for the oxidation of phenol derivatives horseradish peroxidase compound II

 The second-order rate constants for the oxidation of a series of phenol derivatives by horseradish peroxidase compound II were compared to computer-calculated chemical parameters characteristic for this reaction step. The phenol derivatives studied were phenol, 4-chlorophenol, 3-hydroxyphenol, 3-methylphenol, 4-methylphenol, 4-hydroxybenzoate, 4-methoxyphenol and 4-hydroxybenzaldehyde. Assuming a reaction of the phenolic substrates in their non-dissociated, uncharged forms, clear correlations (r = 0.977 and r = 0.905) were obtained between the natural logarithm of the second-order rate constants (ln k _app and ln k ₂ respectively) for their oxidation by compound II and their calculated ionisation potential, i.e. minus the energy of their highest occupied molecular orbital [E(HOMO)]. In addition to this first approach in which the quantitative structure-activity relationship (QSAR) was based on a calculated frontier orbital parameter of the substrate, in a second and third approach the relative heat of formation (ΔΔHF) calculated for the process of one-electron abstraction and H^• abstraction from the phenol derivatives was used as a parameter. Plots of the natural logarithms of the second-order rate constants (k _app and k ₂) for the reaction and the calculated ΔΔHF values for the process of one-electron abstraction also provide clear QSARs with correlation coefficients of –0.968 and –0.926 respectively. Plots of the natural logarithms of the second-order rate constants (k _app and k ₂) for the reaction and the calculated ΔΔHF values for the process of H^• abstraction provide QSARs with correlation coefficients of –0.989 and –0.922 respectively. Since both mechanisms considered, i.e. initial electron abstraction versus initial H^• abstraction, provided clear QSARs, the results could not be used to discriminate between these two possible mechanisms for phenol oxidation by horseradish peroxidase compound II. The computer calculation-based QSARs thus obtained for the oxidation of the various phenol derivatives by compound II from horseradish peroxidase indicate the validity of the approaches investigated, i.e. both the frontier orbital approach and the approach in which the process is described by calculated relative heats of formation. The results also indicate that outcomes from computer calculations on relatively unrelated phenol derivatives can be reliably compared to one another. Furthermore, as the actual oxidation of peroxidase substrates by compound II is known to be the rate-limiting step in the overall catalysis by horseradish peroxidase, the QSARs of the present study may have implications for the differences in the overall rate of substrate oxidation of the phenol derivatives by horseradish peroxidase. Received: 29 March 1996 / Accepted: 17 July 1996     

Abbau chlorierter Benzole,Phenole und Cyclohexan-Derivate durch Benzol und Phenol verwertende Bodenbakterien unter aeroben Bedingungen

From soil samples of different origin (field, grassland and forest soils) small numbers ofNocardin andPseudomonas spec., able to utilize benzene and phenol could be isolated. Organisms which could only utilize phenol and phenolcarboxylic acids were more numerous and consisted mainly ofArthrobacter spec. It was tested to what extent these organisms could also utilize chlorinated aromatic and cyclohexane derivatives. For the degradation studies the bacteria were precultivated on benzene or p-hydroxybenzoic acid and then the compounds used were added. These compounds were labeled by¹⁴C and their degradation rates determined by measuring the¹⁴CO₂ release.Pseudomonas andNocardia spec. precultivated on benzene could also degrade the chlorinated derivatives of benzene and phenol. The monochlorinated derivates were degraded more easily than the di- and trichlorinated derivates. The chlorinated benzenes, especially in higher concentrations, were less degraded than the chlorinated phenols, but with lower concentrations their degradation rates were about similar. This was due to a higher toxicity of the benzenes. The phenol utilizingArthrobacter spec. were only able to degrade phenol and the chlorinated phenols. Benzoic and m-chlorobenzoic acid were degraded to CO₂ by thePseudomonas andNocardia spec. only. The benzene utilizing pseudomonads released more CO₂ from γ-pentachlorocyclohexane than from γ-hexachlorocyclohexane, but none from cyclehexane. Upon precultivation of benzene utilizing pseudomonads in glucose, the aromatic compounds were also degraded, but especially the chlorinated derivatives to a lower extent. In comparison with these soil organisms in pure culture, experiments with soil samples showed a degradation of all compounds which were used by the isolated organisms after variable induction periods. Cyclohexane was degraded slowly to CO₂ by the mixed soil flora in contrast to the benzene or phenol utilizing pure cultures.     

Anaerobic degradation of phenol via carboxylation to 4-hydroxybenzoate: in vitro study of isotope exchange between 14CO2 and 4-hydroxybenzoate

Extracts of denitrifying bacteria grown anaerobically with phenol and nitrate catalyzed an isotope exchange between ¹⁴CO₂ and the carboxyl group of 4-hydroxybenzoate. This exchange reaction is ascribed to a novel enzyme, phenol carboxylase, initiating the anaerobic degradation of phenol by para-carboxylation to 4-hydroxybenzoate. Some properties of this enzyme were determined by studying the isotope exchange reaction. Phenol carboxylase was rapidly inactivated by oxygen; strictly anoxic conditions were essential for preserving enzyme activity. The exchange reaction specifically was catalyzed with 4-hydroxybenzoate but not with other aromatic acids. Only the carboxyl group was exchanged; [U-¹⁴C]phenol was not exchanged with the aromatic ring of 4-hydroxybenzoate. Exchange activity depended on Mn²⁺ and inorganic phosphate and was not inhibited by avidin. Ortho-phosphate could not be substituted by organic phosphates nor by inorganic anions; arsenate had no effect. The pH optimum was between pH 6.5–7.0. The specific activity was 100 nmol ¹⁴CO₂ exchange · min^-1 · mg^-1 protein. Phenol grown cells contained 4-hydroxybenzoyl CoA synthetase activity (40 nmol · min^-1 · mg^-1 protein). The possible role of phenol carboxylase and 4-hydroxybenzoyl CoA synthetase in anaerobic phenol metabolism is discussed.     

Anaerobic degradation of phenol by pure cultures of newly isolated denitrifying pseudomonads

From various oxic or anoxic habitats several strains of bacteria were isolated which in the absence of molecular oxygen oxidized phenol to CO₂ with nitrate as the terminal electron acceptor. All strains grew in defined mineral salts medium; two of them were further characterized. The bacteria were facultatively anaerobic Gramnegative rods; metabolism was strictly oxidative with molecular oxygen, nitrate, or nitrite as electron acceptor. The isolates were tentatively identified as pseudomonads. Besides phenol many other benzene derivatives like cresols or aromatic acids were anaerobically oxidized in the presence of nitrate. While benzoate or 4-hydroxybenzoate was degraded both anaerobically and aerobically, phenol was oxidized under anaerobic conditions only. Reduced alicyclic compounds were not degraded. Preliminary evidence is presented that the first reaction in anaerobic phenol oxidation is phenol carboxylation to 4-hydroxybenzoate.     

Lead optimization of a dihydropyrrolopyrimidine inhibitor against phosphoinositide 3-kinase (PI3K) to improve the phenol glucuronic acid conjugation

Our lead compound for a phosphoinositide 3-kinase (PI3K) inhibitor (1) was metabolically unstable because of rapid glucuronidation of the phenol moiety. Based on structure–activity relationship (SAR) information and a FlexSIS docking simulation score, aminopyrimidine was identified as a bioisostere of phenol. An X-ray structure study revealed a hydrogen bonding pattern of aminopyrimidine derivatives. Finally, aminopyrimidine derivatives 33 showed strong tumor growth inhibition against a KPL-4 breast cancer xenograft model in vivo.     

Phenol derivatives as enhancers and inhibitors of luminol–H2O2–horseradish peroxidase chemiluminescence

Systematic studies on phenol derivatives facilitates an explanation of the enhancement or inhibition of the luminol–H₂O₂–horseradish peroxidase system chemiluminescence. Factors that govern the enhancement are the one-electron reduction potentials of the phenoxy radicals (PhO^•/PhOH) vs. luminol radicals (L^•/LH⁻) and the reaction rates of the phenol derivatives with the compounds of horseradish peroxidase (HRP-I and HRP-II). Only compounds with radicals with a similar or greater reduction potential than luminol at pH 8.5 (0.8 V) can act as enhancers. Radicals with reduction potentials lower than luminol behave in a different way, because they destroy luminol radicals and inhibit chemiluminescence. The relations between the reduction potential, reaction rates and the Hammett constant of the substituent in a phenol suggest that 4-substituted phenols with Hammett constants (σ) for their substituents similar or greater than 0.20 are enhancers of the luminol–H₂O₂–horseradish peroxidase chemiluminescence. In contrast, those phenols substituted in position 4 for substituents with Hammett constants (σ) lower than 0.20 are inhibitors of chemiluminescence. On the basis of these studies, the structure of possible new enhancers was predicted. © 1998 John Wiley & Sons, Ltd.     

Modulation of FAD-dependent monooxygenase activity from aromatic compounds-degrading Stenotrophomonas maltophilia strain KB2

The purpose of this study was purification and characterization of phenol monooxygenase from Stenotrophomonas maltophilia strain KB2, enzyme that catabolises phenol and its derivatives through the initial hydroxylation to catechols. The enzyme requires NADH and FAD as a cofactors for activity, catalyses hydroxylation of a wide range of monocyclic phenols, aromatic acids and dihydroxylated derivatives of benzene except for catechol. High activity of this monooxygenase was observed in cell extract of strain KB2 grown on phenol, 2-methylphenol, 3-metylphenol or 4-methylphenol. Ionic surfactants as well as cytochrome P450 inhibitors or 1,4-dioxane, acetone and n-butyl acetate inhibited the enzyme activity, while non-ionic surfactants, chloroethane, ethylbenzene, ethyl acetate, cyclohexane, and benzene enhanced it. These results indicate that the phenol monooxygenase from Stenotrophomonas maltophilia strain KB2 holds great potential for bioremediation.     

New 3-O-substituted xanthone derivatives as promising acetylcholinesterase inhibitors

A new series of 3-O-substituted xanthone derivatives were synthesised and evaluated for their anti-cholinergic activities against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The results indicated that the xanthone derivatives possessed good AChE inhibitory activity with eleven of them (5, 8, 11, 17, 19, 21-23, 26-28) exhibited significant effects with the IC₅₀ values ranged 0.88 to 1.28 µM. The AChE enzyme kinetic study of 3-(4-phenylbutoxy)-9H-xanthen-9-one (23) and ethyl 2-((9-oxo-9H-xanthen-3-yl)oxy)acetate (28) showed a mixed inhibition mechanism. Molecular docking study showed that 23 binds to the active site of AChE and interacts via extensive π–π stacking with the indole and phenol side chains of Trp86 and Tyr337, besides the hydrogen bonding with the hydration site and π–π interaction with the phenol side chain of Y72. This study revealed that 3-O-alkoxyl substituted xanthone derivatives are potential lead structures, especially 23 and 28 which can be further developed into potent AChE inhibitors.