P-hydroxy Group Research Articles

Structural Insights into Partial Activation of the Prototypic G Protein-Coupled Adenosine A2A Receptor.

The adenosine A2A receptor (A2AAR) belongs to the rhodopsin-like G protein-coupled receptor (GPCR) family, which constitutes the largest class of GPCRs. Partial agonists show reduced efficacy as compared to physiological agonists and can even act as antagonists in the presence of a full agonist. Here, we determined an X-ray crystal structure of the partial A2AAR agonist 2-amino-6-[(1H-imidazol-2-ylmethyl)sulfanyl]-4-p-hydroxyphenyl-3,5-pyridinedicarbonitrile (LUF5834) in complex with the A2AAR construct A2A-PSB2-bRIL, stabilized in its inactive conformation and being devoid of any mutations in the ligand binding pocket. The determined high-resolution structure (2.43 Å) resolved water networks and crucial binding pocket interactions. A direct hydrogen bond of the p-hydroxy group of LUF5834 with T883.36 was observed, an amino acid that was mutated to alanine in the most frequently used A2AAR crystallization constructs thus preventing the discovery of its interactions in most of the previous A2AAR co-crystal structures. G protein dissociation studies confirmed partial agonistic activity of LUF5834 as compared to that of the full agonist N-ethylcarboxamidoadenosine (NECA). In contrast to NECA, the partial agonist was still able to bind to the receptor construct locked in its inactive conformation by an S913.39K mutation, although with an affinity lower than that at the native receptor. This could explain the compound's partial agonistic activity: while full A2AAR agonists bind exclusively to the active conformation, likely following conformational selection, partial agonists bind to active as well as inactive conformations, showing higher affinity for the active conformation. This might be a general mechanism of partial agonism also applicable to other GPCRs.

Synthesis and Crystal Structure Analysis of Some Aromatic Imines of Syringaldehyde

A series of syringaldehyde imines with para-substituted anilines have been synthesized in a good yield, and their crystal structures have been analyzed. The orientation of the syringaldehyde hydroxyl group plays in important role in the intermolecular hydrogen-bonding pattern of the molecules. The O–H…N hydrogen bonding interactions primarily determine the three-dimensional packing of the molecules, even though they make up a relatively small percentage of intermolecular interactions in the molecules. The three structures with the p-hydroxy group cis to the imine group give hydrogen-bonded zigzag chains in the monoclinic crystals, while the structure with a trans hydroxy group crystallize in a hexagonal space group (R3¯) and form hydrogen-bonded hexamers. The hexagonal structure also displays Br…Br interactions, forming additional hexameric clusters. The analysis of published p-hydroxyphenyl imine crystal structures from the Cambridge Crystallographic Database revealed patterns in the length of the hydrogen bonding interactions based on steric congestion around the hydroxyl group.

Dual role of arsenite in hydrolysis and post-hydrolysis fluorescence sensing of selective pH-dependent probes.

In comparison to the sensing activity, the reactivity of arsenite (AsO2-) is less explored. Herein, we focused on AsO2- reactivity studies based on its pKa and compared the study with other common anions. All three pKa values of arsenite are >9.0, affording a flexible working pH range to design a probe for reactivity studies. We designed and synthesized six pH dependent benzothiazole-based Schiff bases, namely, 1-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)naphthalen-2-ol (1), 5-(diethylamino)-2-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)phenol (2), 9-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)-2,3,6,7-tetrahydro-1H,5H-pyrido[3,2,1-ij]quinolin-8-ol (3), 5-methoxy-2-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)phenol (4), 4-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)benzene-1,3-diol (5), and 2-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)phenol (6), as probes for hydrolysis studies containing 5% water in acetonitrile. In spite of the presence of water in the solution, no hydrolysis was observed for all the probes in the absence of a salt. In the presence of selected sodium salts of various anions in solution, intramolecular charge transfer (ICT) was observed after the deprotonation of an aromatic hydroxy group at the ortho position with respect to the imine groups within the probes. Among the studied anions, selective AsO2- induced imine hydrolysis was observed for probes 1 and 4. In the case of 5 with both o- and p-hydroxy groups, no hydrolysis was observed in the presence of AsO2-. Probe 6 with only the o-hydroxy group showed very fast hydrolysis with poor selectivity. The p-hydroxy group in 4-(((6-nitrobenzo[d]thiazol-2-yl)imino)methyl)phenol (7) resulted in poor AsO2- induced hydrolysis. The aldehyde, which was generated after hydrolysis of probe 1, showed selective emission at 450 nm in the presence of AsO2-. The time dependent hydrolysis reaction of probe 1 controls the emission intensity enhancement.

Identification and Structure-Activity Relationship of Recovered Phenolics with Antioxidant and Antihyperglycemic Potential from Sugarcane Molasses Vinasse.

Sugarcane molasses vinasse is the residue of the fermentation of molasses and the water and soil environmental pollutants from distilleries. However, its recycling value has been neglected. The chemical analysis of the molasses vinasse led to the isolation of a new benzoyl chloride called 2,3,4-trihydroxy-5-methoxy benzoyl chloride, as well as thirteen known compounds, including six benzoic acids. The structure of the new benzoyl chloride was elucidated on the basis of extensive spectroscopic analysis. The antioxidant activity of all isolated compounds was measured using the ORAC assay. Moreover, we compared the cellular antioxidant activity (CAA) and inhibitory activity against α-amylase and α-glucosidase for structure–activity analysis. The results showed that only vanillic acid had CAA (8.64 μmol QE/100 μmol in the no PBS wash protocol and 6.18 μmol QE/100 μmol in the PBS wash protocol), although other benzoic acid derivatives had high ORAC values ranging between 1879.9 and 32,648.1 μmol TE/g. Additional methoxy groups at the ortho-positions of the p-hydroxy group of benzoic acids enhanced the inhibition of α-glucosidase but reduced the ORAC activity unless at the para-position. This work indicated that phenolics, especially phenolic acids in the sugarcane molasses vinasse, possessed potential antioxidant and antihyperglycemic activity, which improved the utilization rate of resources and reduced the discharge of pollutants.

Curcumin activates G protein-coupled receptor 97 (GPR97) in a manner different from glucocorticoid

Curcumin is a yellow pigment in turmeric (Curcuma longa) with various physiological effects in the body. To elucidate the molecular mechanisms by which bioactive compounds exert their function, identification of their molecular targets is crucial. In this study, we show that curcumin activates G protein-coupled receptor 97 (GPR97). Curcumin dose-dependently activated serum-response element-, but not serum-response factor-response element-, nuclear factor of activated T-cell-response element-, or cAMP-response element-, mediated transcription in cells overexpressed with GPR97. The structure–activity relationship indicated that (i) the double-bonds of the central 7-carbon chain were essential for activation; (ii) a methoxy group on the aromatic ring was required for maximal activity; (iii) the addition of glucuronic acid moiety or a methoxy group to the aromatic ring, but not the methylation of the aromatic p-hydroxy group, eliminated the activity; (iv) the stability of curcumin would be related to receptor activation. Both mutant GPR97(T250A) lacking the cleavage at GPCR proteolysis site and mutant GPR97(ΔN) lacking the N-terminal extracellular region were activated by curcumin and its related compounds similar to wild-type GPR97. In contrast, the synthetic glucocorticoid beclomethasone dipropionate and l-Phe activated wild-type GPR97 and GPR97(T250A), but not GPR97(ΔN). Moreover, curcumin exerted an additive effect on the activation of wild-type GPR97 with beclomethasone dipropionate, but not with l-Phe. Taken together, these results indicate that curcumin activates GPR97 coupled to Gi/Go subunit, and suggest that curcumin and glucocorticoid activate GPR97 in a different manner.

Green fluorescent protein chromophore derivatives as a new class of aldose reductase inhibitors

A number of (Z)-4-arylmethylene-1H-imidazol-5(4H)-ones, which are related to the fluorescent chromophore of the Aequorea green fluorescent protein (GFP), have been synthesized and evaluated their in vitro inhibitory activity against recombinant human aldose reductase for the first time. The GFP chromophore model 1a, with a p-hydroxy group on the 4-benzylidene and a carboxymethyl group on the N1 position, exhibited strong bioactivity with an IC50 value of 0.36 μM. This efficacy is higher than that of sorbinil, a known highly potent aldose reductase inhibitor. Compound 1h, the 2-naphtylmethylidene analogue of 1a, exhibited the best inhibitory effect among the tested copounds with an IC50 value of 0.10 μM. Structure-activity relationship studies combined with docking simulations revealed the interaction mode of the newly synthesized inhibitors toward the target protein as well as the structural features required to gain a high inhibitory activity. In conclusion, the GFP chromophore model compounds synthesized in this study have proved to be potential drugs for diabetic complications.

1,2,3,4,6-Pentakis[-O-(3,4,5-trihydroxybenzoyl)]-α,β-D-glucopyranose (PGG) analogs: design, synthesis, anti-tumor and anti-oxidant activities

1,2,3,4,6-Pentakis[-O-(3,4,5-trihydroxybenzoyl)]-α,β-D-glucopyranose (PGG) 12 has been reported for its antioxidant activities, where the free OH groups in PGG seem to be critical for activities. To explore PGG-based compounds as chemotherapeutic agents and to analyze the contribution of specific OH groups in PGG for anti-cancer activities, we designed and synthesized a series of 27 benzoic and cinnamic acid analogs of PGG. These analogs were tested for cytotoxicities against two human lung (A549 and H1299) and two human colon (HCT116 and HT29) cancer cell lines. Compound 12 (PGG) had highest cytotoxicities against HCT116 and A549 cells with IC50 of 1.61 µM and 3.02 µM, respectively. In contrast, the compound 16 (1,2,3,4,6-pentakis[-O-(4-hydroxy-3-methoxybenzoyl)]-α,β-D-glucopyranose, PVG) was most effective at killing HT29 and H1299 cells with IC50 of 1.76 µM and 3.65 µM, respectively, indicating the mutual contribution of m-methoxy and p-hydroxy groups to the observed cytotoxicities. Moreover, cinnamic acid analogs were less active than the benzoic acid analogs evidenced by higher IC50 values. Furthermore, in cinnamic acid analogs the hydrogenation of double bond to saturated 2-C side chain enhance the cytotoxicities in all four cell lines. Compounds also possess good anti-oxidant and reducing activities. Compound 12 and 26 show the highest antioxidant and reducing activities.

Anti-inflammatory phenolics isolated from Juniperus rigida leaves and twigs in lipopolysaccharide-stimulated RAW264.7 macrophage cells

Inflammation is an essential host defense system particularly in response to infection and injury; however, excessive or undesirable inflammatory responses contribute to acute and chronic human diseases. A high-throughput screening effort searching for anti-inflammatory compounds from medicinal plants deduced that the methanolic extract of Juniperus rigida S. et L. (Cupressaceae) inhibited significantly nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Activity-guided fractionation and isolation yielded 13 phenolic compounds, including one new phenylpropanoid glycosides, 3,4-dimethoxycinnamyl 9-O-β-D-glucopyranoside (1). Among the isolated compounds, phenylpropanoid glycosides with p-hydroxy group (2, 4) and massoniaside A (7), (+)-catechin (10), amentoflavone (11) effectively inhibited LPS-induced NO production in RAW264.7 cells.

Assessment of Enzymatic Methods in the δ18O Value Determination of the l-Tyrosine p-Hydroxy Group for Proof of Illegal Meat and Bone Meal Feeding to Cattle

The δ(18)O value of the p-hydroxy group of L-tyrosine depends on the biosynthesis by plants or animals, respectively. In animal proteins it reflects the diet and is therefore an absolute indicator for illegal feeding with meat and bone meal. The aim of this investigation was to perform the positional (18)O determination on L-tyrosine via a one-step enzymatic degradation. Proteins from plants, herbivores, omnivores, and carnivores were characterized by their δ(13)C, δ(15)N, and δ(18)O values, the latter for normalizing the positional δ(18)O values. Their L-tyrosine was degraded by tyrosine phenol lyase to phenol, analyzed as (2,4,6)-tribromophenol. Degradation by tyrosine decarboxylase yielded tyramine. The δ(18)O values of both analytes corresponded to the trophic levels of their sources but were not identical, probably due to an isotope effect on the tyrosine phenol lyase reaction. Availability of the enzyme, easy control of the reaction, and isolation of the analyte are in favor of tyrosine decarboxylase degradation as a routine method.

Anion directed template synthesis of Cu(II) complexes of a N,N,O donor mono-condensed Schiff base ligand: A molecular scaffold forming highly ordered H-bonded rectangular grids

Anion directed, template syntheses of two dinuclear copper(II) complexes of mono-condensed Schiff base ligand Hdipn (4-[(3-aminopentylimino)-methyl]-benzene-1,3-diol) involving 2,4-dihydroxybenzaldehyde and 1,3-diaminopentane were realized in the presence of bridging azide and acetate anions. Both complexes, [Cu 2(dipn) 2(N 3) 2] ( 1) and [Cu 2(dipn) 2(OAc) 2] ( 2) have been characterized by X-ray crystallography. The two mononuclear units are joined together by basal–apical, double end-on azido bridges in complex 1 and by basal–apical, double mono-atomic acetate oxygen-bridges in 2. Both complexes form rectangular grid-like supramolecular structures via H-bonds connecting the azide or acetate anion and the p-hydroxy group of 2,4-dihydroxybenzaldehyde. Variable-temperature (300–2 K) magnetic susceptibility measurements reveal that complex 1 has antiferromagnetic coupling ( J = −2.10 cm −1) through the azide bridge while 2 has intra-dimer ferromagnetic coupling through the acetate bridge and inter-dimer antiferromagnetic coupling through H-bonds ( J = 2.85 cm −1, J′ = −1.08 cm −1).

Synthesis, Analgesic, Anti-inflammatory and Antimicrobial Activities of Some Novel Pyrazoline Derivatives

Purpose: Microbial infections often produce pain and inflammation. Chemotherapeutic, analgesic and anti-inflammatory drugs are prescribed simultaneously in normal practice. The compound possessing all three activities is not common.The purpose of the present study was to examine whether molecular modification might result in detection of new potential antirheumatic drugs having antimicrobial activities. Method: A series of novel 4-(5 �-substituted aryl-4�, 5 �-dihydropyrazole-3�-yl-amino) phenols 2a-f have been synthesized by treating substituted aryl-N-chalconyl amino phenols 1a-f with hydrazine hydrate. The starting materials were synthesized from p-aminoacetophenone. Their structures were confirmed by IR, 1 H NMR spectral data. The synthesized compounds were investigated for analgesic, antinflammatory and antimicrobial activities. Result: The data reported in Tables 2, 3 & 4 shows that effect of variation in chemical structure on activity was rather unpredictable. Seldom did a particular structural modification lead to uniform alteration in activity in all tests. The substitution which appeared to be most important for high order of activity in the greatest number of test was the p-choloroaryl group. The introduction of p-nitro and phydroxy group in aryl moiety of the pyrazole analogs 2c and 2e produce compounds with potent analgesic, anti-inflamatory and, in a few cases, antimicrobial properties. Conclusion: The observed increase in analgesic, anti-inflammatory and antimicrobial activities are attributed to the presence of 4-NO 2, 2-OH and 4-Cl in phenyl ring at 5-position of pyrazoline ring of synthesized compounds. In some cases their activities are equal or more potent than the standard drugs.

Phenylglycine as a novel P2 scaffold in hepatitis C virus NS3 protease inhibitors

Molecular modeling and inhibitory potencies of tetrapeptide protease inhibitors of HCV NS3 proposed phenylglycine as a new promising P2 residue. The results suggest that phenylglycine might be capable of interacting with the NS3 (protease-helicase/NTPase) in ways not possible for the common P2 proline-based inhibitors. Thus, a series of tripeptides, both linear and macrocyclic, based on p-hydroxy-phenylglycine in the P2 position were prepared and their inhibitory effect determined. When the p-hydroxy group was replaced by methoxy, isoquinolin-, or quinolinyloxy functions, inhibitors with improved potencies were obtained. The P2 phenylglycine-based inhibitors were further optimized by C-terminal extension to acyl sulfonamides and by P1–P3 cyclization, which gave products with inhibition constants in the nanomolar range (∼75 nM).

Regiospecific Aryl Nitration of meso-Tetraarylporphyrins: The Directive Effect of para-Substituent

Regiospecific nitration of single p-substituted tetraphenylporphyrins with excess nitric acid is reported. The directive effects of different p-substituents, including nitro, acetamido and hydroxy groups, were demonstrated. The electron-withdrawing p-nitro group was found to lead to predominant nitration at the neighboring phenyl ring, and the electron-releasing p-acetamido and p-hydroxy groups lead to adjacent nitration at the same phenyl ring.

Effect of carboxylate spacers on the supramolecular self-assembly of dicopper(ii) Schiff base complexes stabilizing water assemblies of different conformations

Dicopper(II) complexes, namely [Cu2L(O2C–CHCH–C6H4-p-OH)]·2H2O (1·2H2O), [Cu2L(O2C–CH2–C6H4-p-OH)]·2H2O (2·2H2O) and [Cu2L(O2C–CH2CH2–C6H4-p-OH)]·0.5H2O (3·0.5H2O), having different carboxylate ligands with a p-hydroxyphenyl moiety and the pentadentate Schiff base N,N′-1,3-diylbis(salicylaldimino)propan-2-ol (H3L) in its trianionic form, were prepared and structurally characterized by X-ray crystallography. The complexes have a dicopper(II) unit with an alkoxo bridge from the Schiff base and the carboxylate, showing a three-atom bridging mode. The metal centres in a square planar CuNO3 coordination geometry are antiferromagnetically coupled in the asymmetrically double-bridged dicopper(II) core. A significant effect of the –CHCH–, –CH2– and –CH2CH2– spacers of the carboxylate ligands on the formation of different supramolecular structures is observed. Complex [Cu2L(O2C–CHCH–C6H4-p-OH)], 1, forms a helical supramolecular structure due to hydrogen-bonding interactions involving the p-hydroxy group of the phenol from the carboxylate and one phenoxo oxygen atom from the Schiff base. The lattice waters form a helical one-dimensional chain, in which alternate water molecules are anchored to the supramolecular host and the chain propagates along the crystallographic 21 screw axis. Complex 2 forms water aggregates of quasi-linear and pseudo-hexameric cyclic chair conformations involving lattice water molecules, and the previously mentioned para OH group phenoxo oxygen atom. Complex 3·0.5H2O shows the formation of a supramolecular one-dimensional chain structure due to hydrogen-bonding interactions between the p-OH group and the phenoxo oxygen atom. Two such supramolecular structures are linked by hydrogen-bonding interactions involving the lattice water. Differential scanning calorimetry (DSC) of 1·2H2O gives two endotherms at 61.5 and 88.5 °C for the loss of the “free” and the “anchored” water molecules, respectively. The overall change of enthalpy per water molecule is ∼36 kJ mol−1. Complex 2·2H2O shows an endotherm at 131 °C with a shoulder at ∼126 °C. The enthalpy change per water molecule is ∼26 kJ mol−1. The reversibility in loss or addition of lattice water molecules and the corresponding effect on the overall structure is probed by X-ray powder diffraction studies.

Glycosylation of resveratrol protects it from enzymic oxidation.

Plant polyphenols, including dietary polyphenols such as resveratrol, are important components in the plant antioxidant and defence systems. They are also known to exert beneficial effects on human health through diet. As they are produced, these polyphenols may be subjected to deleterious enzymic oxidation by the plant polyphenol oxidases. They are generally synthesized as glycosides like 5,4'-dihydroxystilbene-3-O-beta-D-glucopyranoside, the 3-glucoside of resveratrol. The effects of the glycosylation and methylation of the parent resveratrol on its enzymic oxidation were studied. Methyl and glucosyl derivatives were synthesized using simple one-step methodologies. The kinetics of their enzymic oxidation by tyrosinases were defined. Substitution at the p-hydroxy group, by either glucose or methyl, abolished enzymic oxidation by both mushroom and grape tyrosinases. Substitution at the m-hydroxy group with methyl had a small effect, but substitution with glucose resulted in a much lower affinity of the enzymes for the glycoside. We suggest that glycosylation of polyphenols in nature helps to protect these vital molecules from enzymic oxidation, extending their half-life in the cell and maintaining their beneficial antioxidant capacity and biological properties.

Acyclic amides as estrogen receptor ligands: Synthesis, binding, activity and receptor interaction

We have prepared a series of bisphenolic amides that mimic bibenzyl and homobibenzyl motifs commonly found as substructures in ligands for the estrogen receptor (ER). Representative members were prepared from three classes: N-phenyl benzamides, N-phenyl acetamides, and N-benzyl benzamides; in some cases the corresponding thiocarboxamides and sulfonamides were also prepared. Of these three classes, the N-phenyl benzamides had the highest affinity for ER, the N-phenyl acetamides had lower, and the N-benzyl benzamides were prone to fragmentation via a quinone methide intermediate. In the N-phenyl benzamide series, the highest affinity analogues had bulky N-substituents; a CF3 group, in particular, conferred high affinity. The thiocarboxamides bound better than the corresponding carboxamides and these bound better than the corresponding sulfonamides. Binding affinity comparisons suggest that the p-hydroxy group on the benzoate ring, which contributes most to the binding, is playing the role of the phenolic hydroxyl of estradiol. Computational studies and NMR and X-ray crystallographic analysis indicate that the two anilide systems studied have a strong preference for the s-cis or exo amide conformation, which places the two aromatic rings in a syn orientation. We used this structural template, together with the X-ray structure of the ER ligand binding domain, to elaborate an additional hydrogen bonding site on a benzamide system that elevated receptor binding further. When assayed on the individual ER subtypes, ERα and ERβ, these compounds show modest binding affinity preference for ERα. In a reporter gene transfection assay of transcriptional activity, the amides generally have full to nearly full agonist character on ERα, but have moderate to full antagonist character on ERβ. One high affinity carboxamide is 500-fold more potent as an agonist on ERα than on ERβ. This work illustrates that ER ligands having simple amide core structures can be readily prepared, but that high affinity binding requires an appropriate distribution of bulk, polarity, and functionality. The strong conformational preference of the core anilide function in all of these ligands defines a rather rigid geometry for further structural and functional expansion of these series.

Hydrogen-Bonded Organic Ferromagnet

Nitronyl nitroxide derivatives carrying a hydroquinone or a resorcinol moiety 2-(2‘,5‘-dihydroxyphenyl)-4,4,5,5-tetramethyl-4,5-dihydro-1H-imidazolyl-1-oxyl-3-oxide (HQNN) and 2-(3‘,5‘-dihydroxyphenyl)-4,4,5,5-tetramethyl-4,5-dihydro-1H-imidazolyl-1-oxyl 3-oxide (RSNN)) have been designed and prepared. These compounds were found to afford hydrogen-bonded crystals. In the case of HQNN, two phases of crystals (α- and β-HQNN) were obtained. Of these, the crystal structure of α-HQNN is characterized by the intramolecular hydrogen bond between the o-hydroxy group and the nitronyl nitroxide and by the intermolecular one-dimensional hydrogen-bonded chain between the o- and p-hydroxy groups of the neighboring molecules, respectively. Two of the hydrogen-bonded chains run in parallel, and they are connected by the bifurcated hydrogen bonds formed between the o-hydroxy groups of the facing molecules in the individual chain. The χT value of this crystal increased monotonously with lowering temperature (ST model, J =...

X-Ray Crystal Structures and Chemical Properties of Heterocyclic Azo Compounds Coupled with m-Alkoxyphenols

Abstract X-Ray crystal structures have been determined for two compounds obtained by the coupling reaction of 2-pyridinediazonium salt and m-methoxyphenol, viz., 3-methoxy-4-(2-pyridylazo)phenol (1) and 5-methoxy-2-(2-pyridylazo)phenol (2). The 2-pyridylazo and 2-thiazolylazo compounds having o-methoxy and p-hydroxy groups, such as 1, have lower pKa values and lower affinities for or higher selectivities to metal ions than corresponding o-hydroxy isomers. The resulting bis complexes with metal ions show molar absorptivities greater than 105.

Purification, sequencing and characterization of single amino acid-substituted phospholipase A2 isozymes from Trimeresurus Gramineus (green habu snake) venom

T. Fukagawa, T. Nose, Y. Shimohigashi, T. Ogawa, N. Oda, K. Nakashima, C.-C. Chang and M. Ohno. Purification, sequencing and characterization of single amino acid-substituted phospholipase A2 isozymes from Trimeresurus gramineus (green habu snake) venom. Toxicon31, 957–967, 1993.—Two phospholipases A2 named PLA2-III and IV were newly isolated from Trimeresurus gramineus (green habu snake) venom in addition to PLA2-I and II reported previously [Odaet al. (1991) Toxicon29, 157; Fukagawaet al. (1992) Toxicon30, 133]. Their isoelectric points were determined to be about 4.5. PLA2-III and IV exhibited almost unchanged lipolytic activity toward egg-yolk when compared with PLA2-I. The amino acid sequences were determined by sequencing the native proteins and the peptides produced by enzymatic (Achromobacter protease I and clostripain) and chemical (hydroxylamine) cleavages of the S-carboxamidomethylated derivative of the proteins. Both proteins consisted of 122 amino acid residues. When compared with PLA2-I, PLA2-III showed only a single amino acid substitution at the N-terminal position; namely from His to Asn. PLA2-IV also showed a single substitution from Ala to Asp at position 72. It was inferred that these amino acid substitutions between PLA2-I and PLA2-III or IV are due to the single base substitution at the corresponding codons of genes, which might be preserved independently. The unique presence of Phe at position 28, where Tyr is commonly located and assumed to be a part of the Ca2+-binding loop, was conserved in both PLA2-III and IV as in PLA2-I. There was no significant difference in the dissociation constants (4.3–5.2 × 10−4 M) for Ca2+ between these PLA2s and Tyr-28-containing PLA2s. These results suggested that the p-hydroxy group of Try-28 does not play a crucial role in binding of PLA2s to Ca2+.

Synthese und Antitumoraktivität von [1,2-Diamino-1-(p-hydroxy)benzyl]ethan(dichloro)platin(II)-Komplexen<br />Synthesis and Antitumor Activity of [1,2-Diamino-1-(p-hydroxy)benzyl]ethane(dichloro)platinum(II) Complexes

The complexes [1,2-diamino-l-(p-hydroxy)benzyl-1-(methyl, ethyl, and phenyl)]ethane-(dichloro)platinum(II) were synthesized and characterized. The benzylic p-hydroxy group of the 1,2-diaminoethane ligand leads to good water-solubility of the complexes. The ligands were synthesized in a Strecker-type reaction, starting from the corresponding p-methoxybenzylketones. The amino nitriles were reduced to the diamines. Finally, the p-methoxy groups were converted into hydroxy groups by ether cleavage. For the preparation of the complexes, K2PtCl4 was added to the ligands. The antitumor activity of the platinum(II) compounds was tested towards the human mammary carcinoma cell line MDA-MB 231. The antitumor effect depends on the substituents and increases in the series phenyl, methyl, and ethyl. As a maximum, a growth inhibition value of 77% was obtained.