Fragment-based discovery of 6-substituted isoquinolin-1-amine based ROCK-I inhibitors

Fragment-based NMR screening of a small literature focused library led to identification of a historical thrombin/FactorXa building block, 17A, that was found to be a ROCK-I inhibitor. In the absence of an X-ray structure, fragment growth afforded 6-substituted isoquinolin-1-amine derivatives which were profiled in the primary ROCK-I IMAP assay. Compounds 23A and 23E were selected as fragment optimized hits for further profiling. Compound 23A has similar ROCK-1 affinity, potency and cell based efficacy to the first generation ROCK inhibitors, however, it has a superior PK profile in C57 mouse. Compound 23E demonstrates the feasibility of improving ROCK-1 affinity, potency and cell based efficacy for the series, however, it has a poor PK profile relative to 23A.     

Methyl or ethyl makes the difference: Synthesis and solid-state structure of 9,10-dialkyl-9,10-dihydro-9,10-digallaanthracenes

The donor-free 9,10-dialkyl-9,10-dihydro-9,10-digallaanthracenes 1 (alkyl: methyl) and 2 (alkyl: ethyl) were prepared by reaction of 1,2-di(chloromercurio)benzene with the corresponding trialkylgallium and isolated as colourless air- and moisture sensitive crystalline compounds. In the solid-state structure of 1, two slightly different monomers 1A and 1B are found, which form a dimer 1A?1B held together by “medium” strong gallium arene π-interactions. Further weak π-interactions between 1A and 1A and 1B and 1B constitute a one-dimensional coordination polymer containing strands of the composition [?(1A?1B)?(1B?1A)?]_n. In contrast, compound 2 crystallizes in the form of distinct molecular units without any further intermolecular π-interactions. The molecular units possess D_2d symmetry and are built by strong π-interactions between two digallaanthracene monomers. Two symmetrical aryl group bridges between two gallium atoms are observed for the first time in the subunits of 2. By addition of a Lewis-base (THF, Pyridine) to 2, a monomeric planar digallaanthracene framework is restored, as proven by an X-ray crystal structure analysis of 2 · 2Py. The different structures of 1 and 2 are explained on the basis of steric effects.     

Allosteric modulators of human A2B adenosine receptor

Background

Among adenosine receptors (ARs) the A_2B subtype exhibits low affinity for the endogenous agonist compared with the A₁, A_2A, and A₃ subtypes and is therefore activated when concentrations of adenosine increase to a large extent following tissue damages (e.g. ischemia, inflammation). For this reason, A_2B AR represents an important pharmacological target.

Methods

We evaluated seven 1-benzyl-3-ketoindole derivatives (7–9) for their ability to act as positive or negative allosteric modulators of human A_2B AR through binding and functional assays using CHO cells expressing human A₁, A_2A, A_2B, and A₃ ARs.

Results

The investigated compounds behaved as specific positive or negative allosteric modulators of human A_2B AR depending on small differences in their structures. The positive allosteric modulators 7a,b and 8a increased agonist efficacy without any effect on agonist potency. The negative allosteric modulators 8b,c and 9a,b reduced agonist potency and efficacy.

Conclusions

A number of 1-benzyl-3-ketoindole derivatives were pharmacologically characterized as selective positive (7a,b) or negative (8c, 9a,b) allosteric modulators of human A_2B AR.

General significance

The 1-benzyl-3-ketoindole derivatives 7–9 acting as positive or negative allosteric modulators of human A_2B AR represent new pharmacological tools useful for the development of therapeutic agents to treat pathological conditions related to an altered functionality of A_2B AR.     

A novel glycosyl donor for synthesis of 2-acetamido-4-amino-2,4,6-trideoxy-α-d-galactopyranosides

2-Azido-4-benzylamino-4-N-,3-O-carbonyl-2,4,6-trideoxy-d-galactopyranosyl trichloroacetimidate (14) was conveniently prepared in six steps by regioselective introduction of an N-benzyl carbamate at O-3 of 6-deoxy-d-glucal 6, followed by mesylation at O-4. Intramolecular displacement of the leaving group afforded oxazolidinone 11. Azidonitration of the bicyclic glycal 11 gave the glycosyl nitrate anomers 12 in good yield and stereoselectivity. Hydrolysis of the anomeric nitrates under aqueous conditions gave the pyranose 13, which was easily converted into the imidate 14. Glycosylation of cyclohexanol by 14 gave glycosides 16α and 16β in a ratio of 4:1.     

Perfluoroalkylated derivatives of 6-deoxy-6-ethylamino-d-galactose, 1-deoxy-1-methylamino-d-glucitol, and 1-amino-1-deoxy-d-glucitol: syntheses, hemocompatibility, and effect on perfluorocarbon emulsion

N-Polyfluoroalkyl derivatives of 6-deoxy-6-ethylamino-1,2;3,4-di-O-isopropylidene-α-d-galactopyranose (8-10), 1-deoxy-1-methylamino-d-glucitol (13-15), and 1-amino-1-deoxy-d-glucitol (16-18), all possessing perfluoroalkyl segment, were prepared using nucleophilic epoxide ring opening of 2-[(perfluoroalkyl)methyl]oxiranes 1-3. Co-emulsifying properties and hemolytic activity of the new perfluoroalkylated amphiphiles were tested. Both types of the polyol derivatives 8-10 and 13-18 generally displayed good to excellent co-emulsifying properties on testing on perfluorodecalin/Pluronic F-68 microemulsions. Mono-perfluoroalkylated compounds 8-10 and 13-15 displayed high hemolysis, whereas acyclic bis-perfluoroalkylated compounds 16-18 were non-hemolytic even for short perfluorobutyl segment (16). The properties were generally improving with increasing perfluoroalkyl chain length.     

6-Halogenopyridin-2-olato complexes with the diruthenium(2+) core: Equilibria between head-to-head and head-to-tail structures

The dinuclear bis(6-X-pyridin-2-olato) ruthenium complexes [Ru₂(μ-XpyO)₂(CO)₄(PPh₃)₂] (X = Cl (4B) and Br (5B)), [Ru₂(μ-XpyO)₂(CO)₄(CH₃CN)₂] (X = Cl (6B), Br (7B) and F (8B)) and [Ru₂(μ-ClpyO)₂(CO)₄(PhCN)₂] (9B) were prepared from the corresponding tetranuclear coordination dimers [Ru₂(μ-XpyO)₂(CO)₄]₂ (1: X = Cl; 2: X = Br) and [Ru₂(μ-FpyO)₂(CO)₆]₂ (3) by treatment with an excess of triphenylphosphane, acetonitrile and benzonitrile, respectively. In the solid state, complexes 4B-9B all have a head-to-tail arrangement of the two pyridonate ligands, as evidenced by X-ray crystal structure analyses of 4B, 6B and 9B, in contrast to the head-to-head arrangement in the precursors 1-3. A temperature- and solvent-dependent equilibrium between the yellow head-to-tail complexes and the red head-to-head complexes 4A-7A and 9A, bearing an axial ligand only at the O,O-substituted ruthenium atom, exists in solution and was studied by NMR spectroscopy. Full ¹H and ¹³C NMR assignments were made in each case. Treatment of 1 and 2 with the N-heterocyclic carbene (NHC) 1-butyl-3-methylimidazolin-2-ylidene provided the complexes [Ru₂(μ-XpyO)₂(CO)₄(NHC)], X = Cl (11A) or Br (12A). An XRD analysis revealed the head-to-head arrangement of the pyridonate ligands and axial coordination of the carbene ligand at the O,O-substituted ruthenium atom. The conversion of 11A and 12A into the corresponding head-to-tail complexes was not possible.     

Ecdysteroids and a sucrose phenylpropanoid ester from Froelichia floridana

Phytoecdysteroid glycosides (1-5) and a phenylpropanoid ester of sucrose (6) were isolated from the whole plant of Froelichia floridana, along with eight known compounds including three ecdysteroids (7-9), four flavonoids (10-13), and one phenolic compound (14). Structures were determined using a combination of spectroscopic techniques. Compounds 1, 2 and 6-14 were tested in vitro for their activity against human DNA topoisomerase I. Compound 13 (diosmetin) showed marginal inhibition against topoisomerase I with IC₅₀ of 130 μM in conjunction with low intercalation ability.     

Nonenzymic,biogenetic-like cyclization of a trienic acetal

The stereoselective synthesis of the trans,trans trienic acetal 9 is described. Alkylation of the lithio derivative of 2-methyl-1-hexene-5-yne (10) with ethylene oxide gave the acetylenic alcohol 11, which on reduction with sodium in liquid ammonia afforded the trans diene 12. Alkylation of the sodium enolate of acetylacetone with 13, the mesylate derived from 12, gave the dione 14; chlorination of 14 and deacylation of the resulting chloro dione 15 provided the α-chloro ketone 16. Conversion of 16 to the trans,trans acetal 9 was accomplished by means of a Cornforth olefin synthesis. Thus, stereoselective attack by the Grignard reagent derived from 1-ethylenedioxy-4-chlorobutane afforded from 16 the chlorohydrin 17, which on treatment with methanolic base was converted to the trans epoxide 18. Deoxygenation of 18 to give the trans,trans acetal 9 was accomplished via the intermediacy of the iodohydrin 19.     

Steroidal glycosides from the marine sponge Pandaros acanthifolium

The chemical composition of the Caribbean sponge Pandaros acanthifolium was investigated and led to the isolation of seven new steroidal glycosides namely pandarosides A-D (1, 3, 4 and 6) along with the three methyl esters of pandarosides A, C, and D (2, 5 and 7). Their structures were characterized as 3β-[β-glucopyranosyl-(1→2)-β-glucopyranosyloxyuronic acid]-16-hydroxy-5α,14β-poriferast-16-ene-15,23-dione (1) and its methyl ester (2), 3β-[β-glucopyranosyloxyuronic acid]-16-hydroxy-5α,14β-poriferast-16-ene-15,23-dione (3), 3β-[β-glucopyranosyl-(1→2)-β-glucopyranosyloxyuronic acid]-16-hydroxy-5α,14β-cholest-16-ene-15,23-dione (4) and its methyl ester (5), 3β-(β-glucopyranosyloxyuronic acid)-16-hydroxy-5α,14β-cholest-16-ene-15,23-dione (6) and its methyl ester (7) on the basis of detailed spectroscopic analyses, including 2D NMR and HRESIMS studies. Pandarosides A-D and their methyl esters (1-7) are all characterized by a rare 2-hydroxycyclopentenone D-ring with a 14β configuration. The absolute configuration of the aglycon part of pandaroside A (1) was assigned by comparison between experimental and TDDFT calculated circular dichroism spectra on the more stable conformer.     

Design and synthesis of 3-(azol-1-yl)phenylpropanes as microbicidal spermicides for prophylactic contraception

We designed a series of 25 3-(azol-1-yl)phenylpropanes which yielded 10 compounds (3, 4, 7, 8, 13, 14, 19, 21, 23, 26) that irreversibly immobilized 100% human sperm at 1% (w/v) concentration in 60 s; 12 compounds (8, 9, 15, 16, 19-21, 23-25, 27, 28) that showed potent microbicidal activity at 12.5-50 μg/mL against Trichomonas vaginalis; and 17 compounds (3-11, 13, 15, 19, 21, 23, 26, 28, 30) that exhibited potent anticandida activity with minimum inhibitory concentration (MIC) of 12.5-50 μg/mL. Almost all the compounds exhibited high level of safety towards normal vaginal flora (Lactobacillus) and human cervical (HeLa) cells in comparison to the marketed spermicide nonoxynol-9 (N-9). All the biological activities were evaluated in vitro. Two compounds (4, 8) with good safety profile exhibited multiple (spermicidal, antitrichomonas and anticandida) activities, warranting further lead optimization for furnishing a prophylactic vaginal contraceptive.     

Structure-dependent spectroscopic and redox properties of copper(I) complexes with bidentate iminopyridine ligands

A series of iminopyridine ligands; cyclopropylpyridin-2-ylmethyleneamine (A), cyclopentylpyridin-2-ylmethyleneamine (B), cyclohexylpyridin-2-ylmethyleneamine (C), and cycloheptylpyridin-2-ylmethyleneamine, (D) and their copper(I) complexes, [Cu(L)₂]⁺ (1a-1d) and [Cu(L)(PPh₃)₂]⁺ (2a-2d) have been synthesized and characterized by CHN analyses, ¹H NMR and IR and UV-Vis spectroscopy. Structures of 1a, 1b, 1c and 2a were determined by X-ray crystallography. The coordination polyhedron about the Cu^I center in the complexes is best described as a distorted tetrahedron. The dihedral angles between the least-squares planes of the chelate ligands show considerable variation from 86.1° in 1a to 68.3° in 1b, indicating the importance of packing forces in the crystalline environment. The UV-Vis spectra of the complexes are characterized by first metal to ligand charge transfer bands increasing in wavelength with increasing size of the ring substituents in the ligands, except for the cyclopropyl compounds (1a and 2a), in good agreement with the variation of the dihedral angles between the ligand planes. Cyclic voltammetry of the complexes indicates a quasireversible redox behavior for the complexes. The bulkier ligands (PPh₃) inhibit the geometric distortion within the oxidized form and the redox potentials of complexes 2a-2d are shifted to more positive values, therefore.     

Kuguacins F-S, cucurbitane triterpenoids from Momordica charantia

Chemical investigation of the vines and leaves of Momordica charantia resulted in isolation of fourteen cucurbitane triterpenoids, kuguacins F-S (1-14), including two pentanorcucurbitacins (6 and 7), one octanorcucurbitacin (8), and two trinorcucurbitacins (11 and 12), along with six known analogues. Their structures were elucidated on the basis of extensive spectroscopic and single-crystal X-ray diffraction analyses. Compounds 1-14 exhibited weak anti-HIV-1 activities in vitro.     

Phomoeuphorbins A-D, azaphilones from the fungus Phomopsis euphorbiae

Four azaphilones, named phomoeuphorbins A-D (1-4) were isolated from cultures of Phomopsis euphorbiae, an endophytic fungus isolated from Trewia nudiflora. Structures of 1-4 were established on the basis of spectroscopic analyses, including application of 2D NMR spectroscopic techniques. Phomoeuphorbins A and C exhibited very weak inhibitory activities against HIV replication in C8166 cells in vitro.     

Dopamine release and molecular modeling study of some coumarin derivatives

4-aryl-2-amino-6-(4-hydroxy-2-oxo-2H-chromen-3-yl)-pyridin-3-carbonitrile (1), 4-aryl-2-oxo-6-(4-hydroxy-2-oxo-2H-chromen-3-yl)-pyridin-3-carbonitriles (2a-2c), 3-(6-aryl-1,2,5,6- tetrahydro-2-thioxopyrimidin-4-yl)-4-hydroxy-2H-chromen-2-one (3a, 3b) and pyrazol-3-yl-4-hydroxycoumarin derivatives (4a-4c, 5, 6a, 6b, 7a, 7b, and 8a-8c) were prepared in order to measure their % change dopamine release in comparison to amphetamine as reference, using PC-12 cells in different concentrations. In addition, the molecular modeling study of the compounds into 3BHH receptor was also demonstrated. The calculated inhibition constant (k_i) implemented in the AutoDock program revealed identical correlation with the experimental results to that obtained binding free energy (ΔG_b) as both parameters revealed reasonable correlation coefficients (R²) being 0.51 involving 10 compounds; (1, 2b, 2c, 3a, 3b, 4a, 4b, 6a, and 8c).     

Effect of outer sphere anions on the structure and color of nitrosylpentaamminechromium complex

Three kinds of crystalline compounds containing the nitrosylpentaamminechromium complexes [Cr(NO)(NH₃)₅]²⁺(A) were obtained: chloride ACl₂ (red-orange), chloride perchlorate ACl(ClO₄) (brown), and perchlorate A(ClO₄)₂ (green). The cause of the color change of the complex A with the change of outer sphere anions was sought using X-ray structural data of ACl₂, ACl(ClO₄), and A(ClO₄)₂. Crystal data: ACl₂, orthorhombic, space group Cmcm, a=10.0236 (9) Å, b=9.098 (3) Å, c=10.357(1) Å, V=944.5 (5) ų, Z=4; ACl(ClO₄), tetragonal, space group P4/nmm, a=7.6986 (8) Å, c=9.9566(8) Å, V=590.1 (1) Å^3,Z=2; A(ClO₄)₂, orthorhombic, space group Pnma, a=15.760 (2) Å, b=11.480(2) Å, c=7.920 (2) Å, V=1432.9 (4) ų, Z=4. The complex cation in ACl₂ has a distorted octahedral structure with a linear CrNO moiety. The short CrN (nitrosyl) distance of 1.692 (7) Å indicates the presence of multiple bonding between the chromium atom and the nitrogen atom in the nitrosyl group. The interatomic distances and angles within the complex cations hardly change with the change of the counter anions, while the distances between the complex cations in each crystal increase in the order ACl₂<ACl(ClO₄)<A(ClO₄)₂. The bulky perchlorate anions seems to separate the complex cations, while smaller chloride anions are not large enough to separate them. The distance (3.213(5) Å) between O(NO) and N(NH₃ in the adjacent complex cation) is rather short in the crystal of ACl₂, and there are six hydrogen bonds, where the NO group is surrounded by four NH₃ ligands. The distance (4.002(5) Å) between O(NO) and N(NH₃) is much longer in the crystal of A(ClO₄)₂, indicating the presence of no hydrogen bonding. In the crystal of ACl(ClO₄) the distance (3.452(4) Å) between O(NO) and N(NH₃) is in between those of ACl₂ and A(ClO₄)₂. The presence of hydrogen bonding between O(NO) and N(NH₃ in the adjacent complex cation) seems to cause the color change with the change of outer sphere anions.     

Antitumoral and antileishmanial dioncoquinones and ancistroquinones from cell cultures of Triphyophyllum peltatum (Dioncophyllaceae) and Ancistrocladus abbreviatus (Ancistrocladaceae)

From the methanolic extracts of solid callus cultures from two species of the closely related palaeotropical plant families Dioncophyllaceae and Ancistrocladaceae seven new natural naphthoquinones were isolated, dioncoquinones A (4) and B (5) from Triphyophyllum peltatum, and ancistroquinones B (6), C (7), D (9), E (10), and F (12) from Ancistrocladus abbreviatus. Their structures were elucidated by spectroscopic, chemical, and computational methods. Furthermore, the already known naphthoquinones plumbagin (2), droserone (3), malvone A (8), and nepenthone A (11) were found in the extract of A. abbreviatus. Dioncoquinones A (4) and B (5) showed good - and specific - activity against Leishmania major, while they were not active against other protozoic parasites. Moreover, treatment with 4 and 5 strongly induced apoptosis in human tumor cells derived from two different B cell malignancies, B cell lymphoma and multiple myeloma, without any significant toxicity towards normal peripheral mononuclear blood cells.     

Influence of cis-protecting groups toward ligand exchange reactions in polynuclear Pd(II)-based coordination cages

Polynuclear self-assembly molecules of general formula [{Pd(en)}_x(ligand)_y](NO₃)_2x (A) undergo ligand exchange reaction when heated in DMSO. A mixture of [Pd_m(ligand)_n](NO₃)_2m (B) and [Pd(en)₂](NO₃)₂ (C) is generated in this process. The binuclear compound (A) containing a bidentate, non-chelating ligand 1,4-bis(4′-pyridylmethyl)-2,3,5,6-tetrafluorobenzene, is subjected to ligand exchange where upon the compound (A) remains in a dynamic equilibrium with the mixture of ensuing (B) and (C). Combination of separately prepared (B) and (C) also generates (A), thus equilibrium of (A) with (B) and (C) is again observed. We predict [{Pd(bpy)}_x(ligand)_y](NO₃)_2x (A′) where 2,2′-bipyridyl (bpy) is the cis-protecting group would not probably undergo ligand exchange. The idea was conceived from the fact that (bpy) is more rigid compared to (en) moreover one of the expected products in the event of ligand exchange [Pd(bpy)₂](NO₃)₂ (C′) is not really very stable unlike (C). In fact, when (A′) is heated in DMSO no ligand exchange is observed at all. More interestingly combination of (B) and (C′) generated (A′) smoothly. Mononuclear complexes obtained from the ligand 4-phenylpyridine are also used for similar study for comparison. It is suggested that (bpy) could serve as a better cis-protecting group for Pd(II)-based self-assembly coordination cage compounds particularly when dissolution of the assemblies in polar solvents and heating of the resultant solution is required.     

Synthesis of novel pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives and their inhibition against growth of A549 and H322 lung cancer cells

A series of substituted pyrazolo[1,5-a]pyrazin-4(5H)-one was synthesized by the reaction of ethyl 1-(2-oxo-2-phenylethyl)-3-phenyl-1H-pyrazole-5-carboxylate derivatives and 2-(2-aminoethoxy)ethanol or 2-morpholinoethanamine in the condition of microwave-assisted one-step and solvent-free in a good yield. The structures of the compounds were determined by IR, ¹H NMR and mass spectroscopy. In addition, a representative single-crystal structure was characterized by using X-ray diffraction analysis. Preliminary biological evaluation showed that the compounds could inhibit the growth of A549 and H322 cells in dosage-dependent manners.     

Syntheses and structures of cobalt(III) alcoholate complexes formed by addition of a water molecule across 2-pyridyl substituted imine function

Schiff bases L1-L5 {N-[1-pyridine-2-ylethylidene]pyridine-2-amine (L1), 3-methyl-N-[1-pyridine-2-ylmethylidene]pyridine-2-amine (L2), 3-methyl-N-[1-pyridine-2-ylethylidene]pyridine-2-amine (L3), 4-methyl-N-[1-pyridine-2-ylmethylidene]pyridine-2-amine (L4), 4-methyl-N-[1-pyridine-2-ylethylidene]pyridine-2-amine (L5)} were synthesized and on reaction with Co(NO₃)₂·6H₂O, complexes having the molecular formulae [Co(L1O)₂]NO₃ (1), [Co(L2O)₂]NO₃·xH₂O (2a, x = 2; 2b, x = 3), [Co(L3O)₂]NO₃ (3), [Co(L4O)₂]NO₃·4H₂O (4), [Co(L5O)₂]NO₃ (5) were isolated from the respective imines. The salt [Co(L2O)₂]PF₆ (2c) was obtained by treating 2 with KPF₆. Complexes 1-5 were formed as a result of addition of a water molecule across the imine function and the resultant alcohol binds in its deprotonated form. The alcoholate ion remained bound in a facial tridentate fashion to the low-spin cobalt(III). X-ray crystal structure determination confirmed the presence of trans-trans-trans-N_AN_PO (A = aminopyridyl and P = pyridyl) disposition in 2a and cis-cis-trans-N_AN_PO in 2b, 2c and 4. Water dimers in 2a, 2b, 4 and water-nitrate ion network in 2a were other notable features.     

Chemical constituents from fruits of Harrisonia perforata

Eight limonoids (1-8) including three A, B and D-seco-16-nor-type ones, 5,6-dehydrodesepoxyharperforin C2 (1), harrpernoid B (2), and its C-9S epimer, harrpernoid C (3), along with six known compounds (9-14), were isolated from fruits of Harrisonia perforata. Extensive spectroscopic analysis was used to elucidate their structures and stereochemistries. Further confirmation of structures of 1 and 2 were obtained by single-crystal X-ray diffraction. Limonoids (1-8) were evaluated for their anti-tobacco mosaic virus activity and in vitro cytotoxicity against A549 and HL60 cell lines; only compound 2 showed weak activity.