Synthesis, and QSAR analysis of anti-oncological active spiro-alkaloids.

QSAR study describes the anti-neoplastic spiro-alkaloids with relevant molecular descriptors using CODESSA III software. The dispiro[3H-indole-3,2'-pyrrolidine-3',3"-piperidines] 24-48 were synthesized via [3 + 2]-cycloaddition reaction of azomethine ylides, (generated in situ via decarboxylative condensation of isatins 21-23 with sarcosine) and 3E,5E-1-alkyl-3,5-bis(arylmethylidene)-4-piperidones 10-20. Some of the synthesized analogues exhibited promising antitumor properties against HELA (cervical), HEPG2 (liver), T-47D, MCF7 (breast), and HCT116 (colon) human tumor cell lines, demonstrating activity close to or even better than the standard Doxorubicin, based on in vitro Sulfo-Rhodamine-B bio-assay.

5''-Benzyl-idene-5-chloro-1',1''-dimethyl-4'-phenyl-dispiro-[indoline-3,2'-pyrrolidine-3',3''-piperidine]-2,4''-dione.

The title compound, C30H28ClN3O2, features two spiro links, one connecting the piperidine and pyrrolidine rings, and the other connecting the pyrrolidine ring and indole residue. The configuration about the ethene bond is E. The piperidine ring adopts a half-chair conformation where the C atom connected to the spiro-C atom lies 0.713 (3) Šout of the plane of the remaining five atoms (r.m.s. deviation = 0.086 Å). The pyrrolidine ring has an envelope conformation with the flap atom being the methyl-ene C atom. Centrosymmetric eight-membered {⋯HNCO}2 amide synthons feature in the crystal packing. These are consolidated into a three-dimensional architecture by phen-yl-pyrrolidine C-H⋯N and chloro-benzene-pyrrolidine-bound phenyl C-H⋯π inter-actions.

5-Chloro-5''-(4-chloro-benzyl-idene)-4'-(4-chloro-phen-yl)-1''-ethyl-1'-methyl-dispiro-[indoline-3,2'-pyrrolidine-3',3''-piperidine]-2,4''-dione.

Two spiro links are found in the title compound, C31H28Cl3N3O2, one connecting the piperidine and pyrrolidine rings, and the other connecting the pyrrolidine ring and indole residue. The piperidine ring adopts a half-chair conformation, in which the C atom connected to the spiro-C atom lies 0.741 (3) Šout of the plane of the remaining five atoms (r.m.s. deviation = 0.053 Å). The pyrrolidine ring has an envelope conformation with the flap atom being the methyl-ene C atom. Centrosymmetric eight-membered {⋯HNCO}2 amide dimers are the most significant feature of the crystal packing. These are connected into layers parallel to (-120) by C-H⋯O and π-π inter-actions between pyrrolidine-bound benzene rings [inter-centroid distance = 3.8348 (15) Å]. Slipped face-to-face inter-actions between the edges of pyrrolidine-bound benzene [shortest C⋯C separation = 3.484 (4) Å] connect the layers into a three-dimensional architecture.

5-Chloro-5''-[4-(di-methyl-amino)-benzyl-idene]-4'-[4-(di-methyl-amino)-phen-yl]-1',1''-di-methyl-dispiro-[indoline-3,2'-pyrrolidine-3',3''-piperidine]-2,4''-dione.

The title compound, C34H38ClN5O2, has spiro links connecting the pyrrolidine ring and indole residue, as well as the piperidine and pyrrolidine rings. A half-chair conformation is found for the piperidine ring with the C atom connected to the spiro-C atom lying 0.738 (4) Šout of the plane of the remaining five atoms (r.m.s. deviation = 0.0407 Å). The methyl-ene C atom is the flap in the envelope conformation for the pyrrolidine ring. In the crystal, supra-molecular chains are sustained by alternating eight-membered {⋯HNCO}2 and 14-membered {⋯HC5O}2 synthons. Chains are connected into a three-dimensional network by (pyrrolidine-bound phenyl-meth-yl)C-H⋯π(pyrrolidine-bound phen-yl) edge-to-face inter-actions.

5-Chloro-5''-(4-chloro-benzyl-idene)-4'-(4-chloro-phen-yl)-1',1''-dimethyldi-spiro-[indoline-3,2'-pyrrolidine-3',3''-piperidine]-2,4''-dione.

The racemic title compound, C30H26Cl3N3O2, comprises two spiro links, the first connecting the piperidine and pyrrolidine rings and the other connecting the indole and pyrrolidine rings. The piperidine ring adopts a half-chair conformation, while the pyrrolidine ring has an envelope conformation with the unsubstituted C atom as the flap. The dihedral angles between the two p-Cl-substituted benzene rings and the indole ring are 33.13 (14) and 54.11 (14)°. In the crystal, mol-ecules form inversion dimers through pairs of N-H⋯O hydrogen bonds [graph set R 2 (2)(8)]. Aromatic C-H⋯O hydrogen bonds extend these dimers into a ribbon structure, enclosing R (2) 2(14) ring motifs, along the a-axis direction.

1'-Methyl-4'-(4-methyl-phen-yl)dispiro-[indane-2,3'-pyrrolidine-2',3''-indoline]-1,2''-dione.

In the title mol-ecule, C(27)H(24)N(2)O(2), the pyrrolidin-2-one ring is almost planar (r.m.s. deviation = 0.003 Å), the pyrrolidine ring has an envelope conformation (the N atom is the flap atom) and the cyclo-penta-none ring is twisted about the C(q)-C(m) bond (q = quaternary and m = methylene). The ketone O atoms are directed to opposite sides of the mol-ecule. Supra-molecular chains along the a axis are formed in the crystal packing mediated by N-H⋯N and C-H⋯O inter-actions. These are connected into layers in the ab plane via C-H⋯π inter-actions.

Extraction and phytochemical investigation of Calotropis procera: effect of plant extracts on the activity of diverse muscles.

CONTEXT: Calotropis procera (Ait.) R.Br. (Asclepiadaceae) is a shrub or small tree that grows wild in Egypt. Calotropis acts as a purgative, anthelmintic, anticoagulant, palliative (in problems with respiration, blood pressure), antipyretic, and analgesic, and induces neuromuscular blocking activity. Little research has been done to study the electrophysiological effects of this plant's extracts on cardiac, smooth, and skeletal muscle activities. OBJECTIVE: The present study was conducted to determine the phytochemical composition and the effect of the total alcohol extract of the shoot of the plant, which contains almost all of C. procera's cardiac glycosides, flavonoids, and saponins. Also, this study attempted to throw more light on the electrophysiological effects of the plant extracts on cardiac, smooth, and skeletal muscle activities and to clarify the mechanism(s) of their observed action(s). MATERIALS AND METHODS: The aerial parts of the plant were air dried and their ethanol extracts partitioned with successive solvents. Cardiac, smooth, and skeletal muscles were used in this study to investigate the physiological and pharmacological effects of the plant extracts from different solvents. The data were analyzed by paired t-test. RESULTS: The phytochemical investigation of Calotropis procera revealed the presence of cardenolides, flavonoids, and saponins. The effects of ethanol, n-butanol, and ethyl acetate (EtOAc) extracts were each evaluated on isolated toad heart and their mechanisms of action determined. Perfusion with 2 µg/mL ethanol, 0.2 µg/mL butanol, and 0.2 µg/mL EtOAc extracts caused a significant decrease in heart rate (bradycardia), significant increase in the force of ventricular contraction, and increase in T-wave amplitude. In addition, the effects of different extracts of the studied plant on smooth muscle and skeletal muscle were investigated in this study. The different extracts and latex of C. procera induced a negative chronotropic effect and decreased the heart rate (HR) of isolated toad heart. The different extracts increased the power of contraction of the duodenum (trace a). Pretreatment with atropine sulfate as a muscarinic receptor blocker abolished the stimulatory effect of the different plant extracts and latex of C. procera (trace b). DISCUSSION: The present data suggest that ethanol, butanol, and EtOAc extracts of Calotropis procera have negative chronotropism and positive inotropism. Verapamil could abolish the inotropic effect of ethanol as well as that of butanol and EtOAc extracts. Meanwhile, atropine did not abolish the observed negative chronotropic effect. The ethanol extract increased the power of contraction of rabbit duodenum, but atropine abolished this effect. It also decreased the skeletal muscle contraction; this effect could be through blocking of the nicotinic receptors. Butanol and EtOAc extract data for smooth and skeletal muscles are very close to those for the corresponding ethanol extract of the studied plant. The present data for C. procera indicate its direct action on the myocardium, its increase of smooth muscle motility, and its relaxation of skeletal muscle contraction. The chemical constituents could directly affect the cell membrane probably through receptors coupling to G proteins. They regulate the ion channel physiology as in the myocardium. CONCLUSION: The present data on the extracts of C. procera indicate a direct action on the myocardium, stimulatory effect on smooth muscle motility, and relaxant action on skeletal muscle contraction. Chemical constituents could directly affect the cell membrane probably through receptors coupling to G proteins. They regulate the ion channel physiology as in the myocardium.

The role of mast cells and macrophages in amiodarone induced pulmonary fibrosis and the possible attenuating role of atorvastatin.

Amiodarone (AM) is an effective anti-arrhythmic drug. We investigated the role of mast cells and macrophages on AM induced pulmonary fibrosis and the action of atorvastatin on this fibrosis. Rats were allocated into four groups; negative control (1), positive control (2), 30 mg/kg body weight/day AM (3) and AM + 10 mg/kg/day atorvastatin (4). Lungs were harvested and prepared for histology and immunohistochemistry. Hematoxylin and eosin stained sections of group 3 exhibited disorganized lung architecture. We found cellular debris in the lumen of both intrapulmonary bronchi and bronchioles with partial disruption of the thickened epithelial lining and mononuclear cellular infiltration into the lamina propria. We also observed thickening of the epithelial lining and the smooth muscle layer. Congested, dilated and thickened blood capillaries and thickened inter-alveolar septa were observed with mononuclear cellular infiltrates in the lung of group 3. Most alveoli were collapsed, but some dilated ones were detected. In some alveoli, type ІІ pneumocytes were increased, while type I cells were decreased. We observed significant increases in the amount of collagen in the thickened inter-alveolar septa, around bronchioles and around blood capillaries in sections from group 3. We found a significant increase in mast cells and alveolar macrophages in group 3 compared to group 1. Mast cells and macrophages appear to play important roles in AM induced pulmonary fibrosis. Atorvastatin appears to attenuate this condition.

A Case-control Study of Haemorrhagic Septicaemia in Buffaloes and Cattle in Karachi, Pakistan, in 2012.

A retrospective epidemiological case-control study was performed in Karachi, Pakistan, from January to April 2013. The owners of 217 dairy cattle and buffalo farms from six different locations in Karachi were interviewed. The aim of the study was to identify risk factors associated with the presence of haemorrhagic septicaemia (HS). Farms with a history of at least one instance of sudden death in a dairy animal during 2012 and a positive clinical HS diagnosis (made by local veterinarians) were defined as cases. Farms having no history of sudden deaths in 2012 were defined as controls. Univariable analyses were initially conducted, and factors with P ≤ 0.25 were offered to a multivariable logistic regression model to identify putative risk factors. The final multivariable logistic model contained five factors. Vaccination was found to be a protective factor (OR = 0.22) along with the length of time cattle were kept on farm (months). For every extra month cattle were kept, the odds of HS disease were reduced by a factor of 0.9. In contrast, for every extra animal in a herd, the risk of infection increased by a factor of 1.01. Supplying underground water and the presence of foot and mouth disease on the farm increased the risk by 2.90 and 2.37, respectively. To understand the epidemiology of HS in Karachi dairy herds, more in-depth research is required to study the risk and protective factors identified in this survey and to evaluate risk mitigation strategies, where possible.

Ginkgo biloba extract (EGb 761) modulates bleomycin-induced acute lung injury in rats.

The effect of Ginkgo biloba extract (EGb 761) on bleomycin (BLM)-induced acute lung injury was studied in rats. The responsiveness of isolated pulmonary arterial rings to 5-hydroxytryptamine (5-HT) as well as the levels of some relevant biochemical markers in the lung tissue were taken as evidence for the acute lung injury. BLM was given intraperitoneally at a dose of 15 mg/kg/day for five consecutive days. It was found that BLM treatment attenuated the vasoconstrictor effect of 5-HT on the isolated pulmonary arteries. In lung tissues BLM also elevated the level of lipid peroxides and enhanced the activity of glutathione peroxidase. On the other hand, the level of glutathione and the activity of alkaline phosphatase were reduced. Body weight, lung weight and tissue glutathione-S-transferase activity were, however, not altered. Oral administration of EGb 761 at a dose of 100 mg/kg/day for five consecutive days did not alter any of the chosen biochemical parameters in the lung tissue except for a slight reduction in alkaline phosphatase activity. However, treatment with EGb 761 reduced the responsiveness of the pulmonary artery to 5-HT. Administration of EGb 761 (100 mg/kg/day; po) two hours prior to BLM (15 mg/kg/day; ip), for five consecutive days blunted the occurrence of further reduction in the vasoconstrictor response of the pulmonary artery to 5-HT. Furthermore, EGb 761 tended to normalize BLM-induced alterations in the measured biochemical markers in the lung tissue. The apparent modulatory influence of EGb 761 on BLM-induced acute lung injury stems, at least in part, from its beneficial free radical scavenging properties that provide the extract with antioxidant activity.

Pasteurella haemolytica A2 infection in two sheep flocks in Tripoli area (Libya).

330 sheep from two different flocks were examined for respiratory manifestations including serous to mucopurulent nasal discharge dyspnoea, cough and light depression. 55 animals were sick (16.6%) and 13 died (3.9%). Samples were taken from healthy, diseases and dead cases as well. Pasteurella haemolytica A2 was identified from 18 isolates, and serotyped by rapid plate agglutination (RPA). Drug susceptibility was tested and treatment applied in line with the results.

First observation of camel (Camelus dromedarius) lymphadenitis in Libya. A case report.

The clinical signs of lymphadenitis in camels in Libya were investigated. Four animals 6 to 8 years old were inappetent, emaciated and slightly anemic. The disease was characterized by swelling and abscess formation in the inferior cervical lymph nodes at the base of the neck. Corynebacterium pyogenes was the causative microorganism of this diseased condition.

The inhibition by ADP of NADPH-supported adrenal steroid 11beta-hydroxylation.

The 11beta-hydroxylation of deoxycorticosterone to form corticosterone in adrenal mitochondria has been found to be inhibited by ADP and ATP, with ADP being the more inhibitory of the two. The evidence suggests that the ADP directly affects the enzyme system.

Concerning the subcellular distribution of 3beta-hydroxysteroid dehydrogenase/isomerase in the rat adrenal.

The distribution of 3beta-hydroxysteroid dehydrogenase/isomerase in the subcellular fractions of rat adrenal gland has been determined. Based on the total activity found, 55% was in the microsomal fraction, 10% in the heavy-mitochondrial fraction, 3% in the light-mitochondrial fraction and 26% in the fraction consisting of cell-debris plus nuclei. Ninety-five percent of the total activity was recovered in the fractions. Approximately half the activity in the heavy-mitochondrial fraction could be accounted for by microsomal contamination.

Some characteristics of adrenal steroidogenesis and their possible relationships to the action of the adrenocorticotropic hormone.

PIP: An attempt to define in quantitative terms the characteristics of the biphasic rate curve for pregnenolone synthesis in cell-free systems from the adrenal using male Sprague-Dawley rats is reported. When adrenocorticotropic hormone (ACTH) was used 2 units of .2 ml of .9% saline were injected ip 15 minutes before killing the rats. The effect of ACTH on adrenal steroidogenesis is in the stimulation of the rate of conversion of cholesterol to pregnenolone. This reaction sequence is thought to occur in the mitochondria. Methods of preparing subcellular fractions are described. Incubation of pregnenolone with mitochondria for 20 minutes at 20 degree C resulted in a 70% disappearance of the pregnenolone. This loss does not occur if the mitochondria are boiled, indicating an enzymatic process. The rate of pregnenolone synthesis characteristically shows a biphasic curve with a rapid primary rate and a slower secondary rate. ACTH administration in vivo increased both rates but the percentage increase was greater for the secondary rate. In addition an increase in the duration of the primary rate resulted. Different explanations are offered for these characteristics. Pregnenolone may act as an inhibitor of its own synthesis from cholesterol but not from 20alpha-hydroxycholesterol. Substances that cause mitochondria to swell may stimulate pregnenolone synthesis. Another theory proposes that the limiting ACTH-sensitive step is the rate at which mitochondrial cholesterol is transported to or binds to the cholesterol side-chain cleavage enzyme. The possible role of an inhibitor in the regulation of steroidogenesis is indicated. Data are consistent with the observation that the transition from the primary rate to the slower secondary rate shows the accumulation of an inhibitory substance. The action of ACTH would then be to modify the structure of the cholesterol side-chain cleavage enzyme so that there is a decreased susceptibility of the enzyme to the inhibitor.^ieng

Studies on the stimulation by Ca2+ and the inhibition by ADP of steroid 11beta-hydroxylation in adrenal mitochondria.

The Ca2+ stimulation and ADP inhibition of the steroid 11beta-hydroxylation enzyme system in adrenal mitochondria have been investigated. The Ca2+-stimulation is competitively inhibited by Sr2+. Nucleoside monophosphates were not inhibitory and among the nucleoside diphosphates and triphosphates only ADP, CDP and ATP were inhibitory with ADP being more inhibitory than the other nucleotides. None of the non-inhibitory nucleotides could reverse the ADP inhibition. ADP appears to inhibit the Ca2+-stimulation since the activity in the absence of Ca2+ was not inhibited by ADP. Ca2% and ADP did not affect the binding of deoxycorticosterone to the mitochondrial preparation. Ca2+ stimulated the reduction of cytochrome P-450 by NADPH and this was inhibited by ADP. These effects were independent of the presence of deoxycorticosterone. Neither Ca2+ nor ADP affected the reduction of adrenodoxin by NADPH. The inhibition by ADP with respect to Ca2+ or to deoxycorticosterone is non-competitive. The Ca2+ stimulation with respect to deoxycorticosterone is uncompetitive. Preincubation of the mitochondrial preparation with Ca2+ decreases the inhibition by ADP and preincubation with ADP increases its inhibitory effects. These data have been interpreted to indicate that Ca2+ and ADP influence the relationship between adrenodoxin and cytochrome P-450 so that a complex of increased activity in the reduction of cytochrome P-450 is formed in the presence of Ca2+ and a less active complex is formed in the presence of Ca2+ plus ADP.

Nadolol-glibenclamide interaction: relevance to carbohydrate metabolism in hyperglycaemic rats.

In the present study, hyperglycaemia was induced by intraperitoneal injection of streptozotocin (65 mg/kg). Treatment with glibenclamide (GB) in a dose of 0.45 or 0.9 mg/kg significantly decreased plasma glucose level in a dose-related manner. Administration of nadolol (ND) in a dose of 5 or 10 mg/kg did not affect plasma glucose level. Combined administration of ND (10 mg/kg) with GB (0.45 or 0.9 mg/kg) potentiated the hypoglycaemic effect of GB, an effect prominent 4 hours post-treatment. In relevance to the effect of ND and GB interactions towards other aspects of carbohydrate metabolism, co-administration of the two drugs [ND (10 mg/kg) + GB (0.9 mg/kg)] failed to alter the increase in plasma insulin level and the decrease in blood pyruvate and lactate levels induced by GB alone. Concerning liver glycogen, concurrent administration of the two drugs showed a synergistic effect upon its content (257%), while it was 186% for GB treatment, and 179% for ND alone compared to the hyperglycaemic control value. The data revealed that ND potentiates the hypoglycaemic effect of GB, so it is very important to consider this potentiation when the usage of the combined drug regimens is recommended.

Adverse testicular effects of some quinolone members in rats.

In the last few years, a marked decrease in male fertility has been reported. Environmental factors were recently suspected for this effect. Among those factors is the misuse of drugs and in particular antibiotics. Quinolones are a group of antibacterial agents with broad-spectrum activity. Testicular impairment of some quinolone members is controversial; a matter which stimulated our attention to investigate the adverse testicular effects of the most familiar quinolone members, namely: ofloxacin, ciprofloxacin and pefloxacin. They were given to rats in doses of 72, 135 and 72 mg kg(-1) day(-1) p.o., respectively, for 15 consecutive days. Ofloxacin was also used to establish a dose-response relationship in doses of 36, 72 and 360 mg kg(-1) day(-1) p.o. for 15 consecutive days. Results revealed that ofloxacin, ciprofloxacin and pefloxacin reduced testicular LDH-X activity by 39.8%, 62.7% and 60.7%, respectively. Moreover, sperm count, motility and daily sperm production were markedly decreased. Ofloxacin induced a dose-dependent decrease in testicular LDH-X activity, sperm count and motility. Furthermore, daily sperm production showed a marked reduction which amounted to 26.1% and 40. 0% following administration of ofloxacin (72, 360 mg kg(-1) day(-1) x 15 days), respectively. Moreover, administration of ofloxacin resulted in marked testicular histopathological changes. It is concluded that, ofloxacin, ciprofloxacin and pefloxacin significantly impaired both testicular function and structure in rats.

Effect of bromocriptine on uterine contractility in near-term pregnant rats.

Treatment of pregnant albino rats at gestation day 9 with the dopamine agonist, bromocriptine, in a dose of 0.7 mg kg-1 day-1, i.p. for 11 days produced a significant increase in the normal uterine contractions both in vitro and in vivo. The increase in frequency (F), amplitude (A) and area under the curve (AUC) in the in vitro experiment amounted to 35, 80 and 58%, respectively; while the increase in F and A in the in vivo experiment was 36 and 25%, respectively, in comparison with the corresponding control group. Addition of oxytocin (5x10(-12)-4x10(-11) m) to the uterus isolated from rats pretreated with bromocriptine resulted in marked uterotonic effect (24, 35 and 49% increase in F; 25, 35 and 46% increase in A and 42, 62 and 122% increase in AUC of contractions). Also, the in vivo experiment showed that an injection of oxytocin at the time of investigation (0.125-1.0 I.U. kg-1, i.v.) into rats pretreated with bromocriptine caused a marked increase in F (33, 40 and 81%) and A (33, 37 and 75%) of uterine contractions compared to the values of bromocriptine-treated animals. These results indicate that bromocriptine should be used with caution during pregnancy. In addition, this must be considered when using oxytocin during delivery of females pretreated with bromocriptine.