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Vitamin D3 (vit-D3), as an indispensable and fat-soluble nutrient, is associated with skeletal mineralization and health in mammals. However, such associations have not been well studied in economically important crustaceans. Six levels of vit-D3 with isonitrogenous and isolipidic diets were used to feed Eriocheir sinensis. The range of optimal vit-D3 requirements is 5685.43-10,000 IU/kg based on growth. The crabs fed 9000 IU/kg vit-D3 showed the best growth performance. This vit-D3 dose significantly increased antioxidant capacity in the hepatopancreas and intestine and was optimal for molting and innate immunity via quantitative polymerase chain reaction analysis. Transcriptomics analyses indicate that vit-D3 could alter protein processing in the endoplasmic reticulum, steroid biosynthesis, and antigen processing and presentation. As shown by the enzyme-linked immunosorbent assay, vit-D3 could improve vitamin D receptor, retinoic acid receptor, and C-type lectins concentrations. The 1α,25-dihydroxy vit-D3 content in serum was significantly higher in 3000-9000 IU/kg vit-D3. The study suggests that dietary vit-D3 and its metabolites can regulate molting and innate immunity in crabs.

This study evaluated the effects of dietary myo-inositol (MI) on growth performance, antioxidant status and lipid metabolism of juvenile Chinese mitten crab (Eriocheir sinensis) fed different percentage of lipid. Crabs (4·58 (sem 0·05) g) were fed four diets including a normal lipid diet (N, containing 7 % lipid and 0 mg/kg MI), N with MI supplementation (N + MI, containing 7 % lipid and 1600 mg/kg MI), a high lipid diet (H, containing 13 % lipid and 0 mg/kg MI) and H with MI supplementation (H + MI, containing 13 % lipid and 1600 mg/kg MI) for 8 weeks. The H + MI group showed higher weight gain and specific growth rate than those in the H group. The dietary MI could improve the lipid accumulations in the whole body, hepatopancreas and muscle as a result of feeding on the high dietary lipid (13 %) in crabs. Besides, the crabs fed the H + MI diets increased the activities of antioxidant enzymes but reduced the malondialdehyde content in hepatopancreas compared with those fed the H diets. Moreover, dietary MI enhanced the expression of genes involved in lipid oxidation and exportation, yet reduced lipid absorption and synthesis genes expression in the hepatopancreas of crabs fed the H diet, which might be related to the activation of inositol 1,4,5-trisphosphate receptor (IP3R)/calmodulin-dependent protein kinase kinase-β (CaMKKβ)/adenosine 5’-monophosphate-activated protein kinase (AMPK) signalling pathway. This study demonstrates that MI could increase lipid utilisation and reduce lipid deposition in the hepatopancreas of E. sinensis fed a high lipid diet through IP3R/CaMKKβ/AMPK activation. This work provides new insights into the function of MI in the diet of crustaceans.

The effects of dietary vitamin A level on the growth performance, immunity, oxidation resistance and lipid metabolism of juvenile Chinese mitten crab Eriocheir sinensis (0.84 ± 0.00 g) were determined in the present study. Six group of crabs (four replicate tanks per group with 40 crabs per tank) were respectively fed with 0 (control), 2322, 4511, 8478, 14,333 and 30,444 IU/kg dietary vitamin A for 8 weeks. The weight gain (WG) and specific growth rate (SGR) of juvenile crabs significantly increased in dietary vitamin A supplement groups compared with the control. Dietary vitamin A supplementation increased immunity and antioxidant capacity, hepatopancreas health and alleviated lipid peroxidation in the hepatopancreas of crabs. Moreover, total lipid content in the whole body and hepatopancreas, and triglyceride (TG) content in the hepatopancreas and serum of crabs showed a decreasing tendency with increasing inclusion of vitamin A. The hepatosomatic index (HSI) was lowest in crabs fed 30,444 IU/kg dietary vitamin A. Interestingly, the transcriptome sequencing data showed that the expression of genes related to immunity, antioxidant capacity, lipolysis and β-oxidation was up-regulated. In contrast, the expression of genes related to lipid synthesis was down-regulated in crabs fed 30,444 IU/kg dietary vitamin A compared with the control. This study indicates that dietary vitamin A requirement for E. sinensis is 5340 for WG and 5540 IU/kg for SGR based on the broken-line model.

Copper (Cu) is a widespread pollutant in water and has posed a significant threat to aquatic animals' health. It is essential to find a strategy to reduce the adverse effects of copper on aquatic organisms. This study investigated the role of myo-inositol (MI) in the reducing chronic toxicity induced by copper in Chinese mitten crab (Eriocheir sinensis). Two diets, including 0 mg/kg and 1600 mg/kg MI were fed to crabs exposed to 0 μg/L and 40 μg/L Cu2+ for 7 weeks. Dietary MI could improve the decreased weight gain and specific growth rate induced by copper exposure. Copper exposure increased the copper levels in the whole body and hepatopancreas, increased the malondialdehyde content, and suppressed the antioxidant enzymes system of the crabs. Simultaneously, copper exposure induced DNA fragmentation, decreased the gene expression/activities of immune-related genes/enzymes and the expression of the anti-apoptotic genes, and increased the gene expression of heat shock protein 70 and pro-apoptotic genes. The mitochondrial dysfunction, increased the gene expression of inflammation-related genes and caused the higher activity of damage-related enzymes in the crabs under Cu exposure. The administration of dietary MI markedly ameliorated the copper-induced oxidative stress, immune and hepatopancreas damage, mitochondrial dysfunction, apoptosis and inflammatory responses. This study indicates that dietary MI supplementation can effectively alleviate the negative effect of chronic copper exposure on E. sinensis.

This study investigates the effects of vitamin D3 (VD3) on growth performance, antioxidant capacity, immunity and molting of larval Chinese mitten crab Eriocheir sinensis. A total of 6,000 larvae (7.52 ± 0.10 mg) were fed with six isonitrogenous and isolipidic experimental diets with different levels of dietary VD3 (0, 3000, 6000, 9000, 12000 and 36000 IU/kg) respectively for 23 days. The highest survival and molting frequency were found in crabs fed 6000 IU/kg VD3. Weight gain, specific growth rate, and carapace growth significantly increased in crabs fed 3000 and 6000 IU/kg VD3 compared to the control. Broken-line analysis of molting frequency, weight gain and specific growth rate against dietary VD3 levels indicates that the optimal VD3 requirement for larval crabs is 4825–5918 IU/kg. The highest whole-body VD3 content occurred in the 12000 IU/kg VD3 group, and the 25-dihydroxy VD3 content decreased with the increase of dietary VD3. The malonaldehyde content was lower than the control. Moreover, the superoxide dismutase activity, glutathione peroxidase and total antioxidant capacity of crab fed 6000 IU/kg VD3 were significantly higher than in control. Crabs fed 9000 IU/kg showed the highest survival after 120 h of salinity stress, and the relative mRNA expressions indicate vitamin D receptor (VDR) is the important regulatory element in molting and innate immunity. The molting-related gene expressions showed that the response of crab to salinity was self-protective. This study would contribute to a new understanding of the molecular basis underlying molting and innate immunity regulation by vitamin D3 in E. sinensis.

An 8-wk feeding trial was conducted to evaluate the effects of replacing fishmeal with rapeseed meal (RM) on growth, feed utilization, antioxidant capacity, nonspecific immunity, and growth-related gene expression of Pseudobagrus ussuriensis (initial body weight 14.0 g). Six isonitrogenous and isoenergetic diets were formulated to contain RM to replace fishmeal at 0 (R0), 10 (R10), 20 (R20), 30 (R30), 40 (R40), and 50% (R50), respectively. The results showed that final body weight, specific growth rate (SGR), feed intake, feed efficiency, and protein efficiency ratio decreased with increasing dietary RM levels. Fish fed the diets with 30% or more fishmeal replaced by RM had significantly lower SGR (P

An 8-week feeding trial was conducted to evaluate the effects of replacing fish meal (FM) with cottonseed meal (CM) on growth and immune characteristics of juvenile Ussuri catfish Pseudobagrus ussuriensis. Seven isonitrogenous and isolipidic diets were formulated to contain CM to replace fish meal at 0 (C0, control), 10 (C10), 20 (C20), 30 (C30), 40 (C40), 50 (C50) and 60% (C60), respectively. The replacement level up to 30% did not affect the weight gain and specific growth rate, whereas these parameters were depressed by further replacement level. The apparent digestibility coefficient of dry matter, crude protein and crude lipid decreased with increasing dietary CM level. Fish fed C60 diet had the lowest hepatic total antioxidant capacity but the highest malondialdehyde level. The activities of serum lysozyme, plasma myeloperoxidase and plasma alkaline phosphatase decreased as the proportion of CM increased. Conversely, the inclusion of CM increased the plasma alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase activities. The highest heat shock protein 70 gene expression level was observed in fish fed the C60 diet. In addition, percent mortality increased after challenge with Aeromonas hydrophila. Percent mortality of fish fed the C60 diet was significantly higher than the other groups. Broken-line model analysis based on WG against dietary CM replacement level indicated that the optimum replacement level was 25.3%. However, including too high proportion of CM could depress the antioxidant capacity, non-specific immune response and resistance to A. hydrophila infection of Ussuri catfish.

Effects of dietary Polychlorinated biphenyl (PCB) exposure and dietary vitamin A supplementation on Chinese mitten crab Eriocheir sinensis were studied with the aim to explain dietary PCB toxicity and toxic alleviation by vitamin A intake in crab. Four diets were used including three experimental diets containing 0, 80000 or 240000 IU/kg vitamin A with each experimental diet containing 10 mg PCB/kg diet, and a control diet (without vitamin A and PCB supplementation) in 56 days feeding trial. Crabs fed the PCB-only diet had significantly lower weight gain than those fed the control diet. No significant difference was observed in crab survival among all groups. Crabs fed the PCB-only diet had a significantly higher malondialdehyde content and antioxidase superoxide dismutase activity in the serum and hepatopancreas, and higher erythromycin N-demethylase and glutathione S-transferase activities in the hepatopancreas than those fed the control diet. However, supplementation of dietary vitamin A decreased the levels of all these parameters. The hepatopancreatic cytochrome P450 2 and 4 (CYP2, CYP4), fatty acid binding proteins 3 and 10 (FABP3, FABP10) and intracellular lipolytic enzyme (IL) Messenger Ribonucleic Acid (mRNA) levels in the PCB-only group were significantly higher than those in the control group, and dietary 240000 IU/kg vitamin A supplementation decreased hepatopancreatic CYP4, FABP3, FABP10 and IL enzyme mRNA level. The crabs fed 80000 IU/kg vitamin A supplementation diet had the highest level of retinoid X receptor mRNA in the hepatopancreas. The structure of the hepatopancreas was damaged and the deposit of lipid droplets decreased with dietary PCB exposure. Both levels of vitamin A supplementation alleviated the damage and increased lipid droplets in the hepatopancreas. Dietary PCB exposure significantly reduced total hemocyte count (THC), and phenoloxidase, acid phosphatase activities in the serum. Post-challenge survival of crab in the experimental PCB-only diet group was low compared with that in the control. Supplementation of 240000 IU/kg vitamin A significantly increased the THC and phenoloxidase activity in the serum and post-challenge survival compared with those in the PCB-only group. This study indicates that dietary vitamin A can improve the antioxidant capacity, immune response, detoxification enzymes activities, energy metabolism and hepatopancreas tissue structure of Chinese mitten crab fed PCB contaminated diets.

This study evaluates the effect of dietary supplementation of immunostimulants on the Chinese mitten crab (Eriocheir sinensis) with a single administration of mannan oligosaccharide (MOS), or its combination with either β-glucan or with inulin for 8 weeks. Four diets included an untreated control diet (C), MOS alone (3 g kg−1, M), MOS with β-glucan (3 g kg −1 MOS + 1.5 g kg −1 β-glucan, MB), and MOS with inulin (3 g kg −1 MOS + 10 g kg −1 inulin, MI). The weight gain and specific growth rate of the crabs fed M, MB, and MI diets were improved by lowing feed conversion ratio. The growth and feed utilization of the crabs fed the MB diet were improved compared with the other three groups. The crabs fed the M, MB and MI diets showed a higher intestinal trypsin activity than that in the M and control groups. The highest trypsin activity in the hepatopancreas was observed in the MB group. Crabs fed M, MB and MI diets increased antioxidant system-related enzyme activities, but reduced malondialdehyde. The highest activities of alkaline phosphatase, acid phosphatase, lysozyme and phenol oxidase in the gut and the respiratory burst of the crabs were found in the MB group. The MB diet promoted the mRNA expression of E. sinensis immune genes (ES-PT, ES-Relish, ES-LITAF, p38MAPK and Crustin) compared with the control. After 3 days of infection with Aeromonas hydrophila, the highest survival of crabs was also found in the MB group. This study indicates that the combination of MOS with β-glucan or with inulin can improve growth, antioxidant capacity, non-specific immunity and disease resistance in E. sinensis.

Myo-inositol (MI) as a major intracellular osmolyte can be accumulated to protect cells from a variety of stress. The present study was to evaluate the effects of dietary MI on growth, osmotic response and transcriptional response in the liver of euryhaline fish Oreochromis mossambicus under long-term salinity stress. Five isonitrogenous and isolipidic diets were formulated to include 0 (MI0), 200 (MI200), 400 (MI400), 800 (MI800) and 1600 (MI1600) mg/kg MI. In this study, dietary 400 mg/kg MI significantly increased the weight gain and specific growth rate of O. mossambicus compared with the group without dietary MI supplementation under salinity stress. Moreover, dietary MI decreased serum glucose and insulin levels of fish under long-term salinity stress. Fish fed 400 mg/kg MI supplementation had significantly higher serum cortisol, sodium (Na+) and chloride ion (Cl−) than those fed a diet without MI supplementation. A total of 132.17 million and 115.8 million clean reads were obtained from the 0 mg/kg MI (MI0) and 400 mg/kg (MI400) MI group respectively. Compared with the MI0 group, the expression of 704 genes was significantly changed (521-up and 183-down) in the MI400 group under long-term salinity stress. Transcriptome results indicate that myo-inositol can regulate lipid and pyruvic acid metabolism, increase phospholipid synthesis-related mRNA levels and improve antioxidant properties of O. mossambicus under long-term salinity stress. This study suggests that optimum MI supplementation is 400 mg/kg in the diet based on weight gain against dietary MI and this result provides a nutritional solution for tilapia to cope with salinity stress.

This study was to evaluate the relationship between myo-inositol synthesis and carbohydrate metabolism in Mozambique tilapia (Oreochromis mossambicus). Fish were immediately transferred from 0 psu (practical salinity units, the control) to 20 psu for 96 h with 60 fish in each group. Compared with the control group, the volume of the mitochondria-rich cell in the gills of O. mossambicus was significantly increased by acute hypersaline stress. The osmotic pressure and the contents of Na+, K+ and Cl− in the serum were significantly increased in the 20 psu group. Moreover, fish in the 20 psu group had significantly higher activities of glutathione peroxidase and superoxide dismutase and a higher malondialdehyde content than the control in the liver. Salinity stress significantly increased the serum glucose but decreased the liver glycogen. The activities of glucokinase and phosphofructokinase in the liver and gills were significantly higher in the 20 psu group, indicating that glycolysis could be significantly enhanced by salinity stress. The activities of glucose-6-phosphate synthase and phosphoenolpyruvate carboxykinase in the liver were significantly decreased by salinity stress. The expressions of myo-inositol-1-phosphate synthase and myo-inositol monophosphatase were significantly up-regulated in the gills and kidney. However, the expressions of these two genes were significantly down-regulated in the liver. The myo-inositol content in the gills, kidney and liver in the 20 psu group were significantly increased by salinity stress. This study indicates that salinity stress can impair the gill structure and reduce the antioxidant capacity of O. mossambicus. The changes in MI biosynthesis and carbohydrate metabolism could be an effective strategy to alleviate the adverse effect of salinity stress.

This study investigated the effects of dietary phosphorus (P) on growth, body composition, antioxidant activities and lipid metabolism of juvenile Chinese mitten crab (Eriocheir sinensis). Six diets containing available P levels at 0.20%, 0.44%, 0.99%, 1.37%, 1.75% and 2.01% were prepared and fed to juvenile E. sinensis (0.38 ± 0.01 g) for 56 days. The weight gain, specific growth rate and molting frequency were markedly affected by dietary P, and the highest weight gain and specific growth rate were obtained in crabs fed the 0.99% available P diet. With increasing the dietary available P levels, the whole-body ash, Ca, and P contents, as well as the hepatopancreas and serum P contents increased, while the whole-body Mg content decreased. The increase of dietary P levels significantly decreased the lipid contents of whole-body, hepatopancreas and muscle, while increased the crude protein content of hepatopancreas. The hepatopancreas and serum alkaline phosphatase activities increased with an increase in dietary available P levels from 0.20% to 1.37%, and then decreased afterward. The crabs fed 1.37% available P diet had a markedly higher alkaline phosphatase activity than those fed 0.20%, 0.44% and 2.01% available P diets. Moreover, dietary P significantly affected total antioxidant capacity, the activities of total superoxide dismutase and glutathione peroxidase, and the malondialdehyde content in the hepatopancreas. The highest antioxidant activity and lowest malondialdehyde content in the hepatopancreas were found in the crabs fed 1.37% of available P in the diet. The relative mRNA expression showed that dietary P reduced hepatopancreas lipid accumulation might be produced through suppressing the genes related to lipid synthesis and promoting the genes related to lipid transport. This study demonstrates that optimum dietary P could enhance growth, antioxidant capacity and reduce hepatopancreatic lipid accumulation in juvenile E. sinensis and the range of available P in the diet is suggested to be 1.16%–1.51%.

An 8-week feeding trial was conducted to investigate the impacts of dietary phospholipid (PL) on growth, antioxidant status and lipid metabolism of Eriocheir sinensis juveniles (0.53 ± 0.01 g) fed four diets including high lipid (13%, HL) and low lipid (8%, LL) and each lipid level containing either high PL (5%, HP) or low PL (1%, LP). Final weight, weight gain, specific growth rate and molting frequency in the HL-LL group were lower than those in the HL-HP group. Compared with the HL-LP group, the enzymatic antioxidant activities increased and hepatopancreatic malondialdehyde content decreased in the HL-HP group. In addition, the high dietary PL significantly suppressed the accumulation of lipid in the hepatopancreas and the whole-body induced by high lipid diets. Quantitative real-time PCR showed that the dietary PL reduced excessive lipid deposition in the hepatopancreas through promoting lipid oxidation and exportation as well as inhibiting lipid absorption and synthesis at the transcriptional level. This study indicates that high dietary PL could promote lipid utilization in E. sinensis fed a high-lipid diet by optimizing lipid metabolism to improve growth performance.

The present study explored the mechanisms of gossypol induced inflammatory response in liver tissue of Ussuri catfish (Pseudobagrus ussuriensis), and investigated whether the yeast culture could relieve the inflammatory response of fish. For this purpose, Ussuri catfish were fed two diets with or without yeast culture for 8 weeks, and then fish were treated with 300 mg gossypol/kg body weight by intragastric administration for 15 days. Fish pre-fed diet without yeast culture supplementation and treated with gossypol oral administration (Gos group) had higher interleukin-1β (IL-1β) and interleukin 8 (IL-8) mRNA levels compared with fish pre-fed diet without yeast culture supplementation and treated with sodium carboxymethylcellulose solution oral administration (control group) and pre-fed yeast culture diet and treated with gossypol oral administration (Gos/YC group), but had lower interleukin 10 mRNA level than the control group (P .05) mRNA levels. Besides, the lowest serum lysozyme and alkaline phosphatase activities were observed in fish of Gos group. There was an increased trend in immunoglobulin M content among the groups. Fish in Gos group had significantly lower hepatic catalase, superoxide dismutase and total antioxidant capacity compared with those fish in the control group (P

This study determines the effects of level and source of dietary phospholipid (PL) on growth performance, body composition, antioxidant capacity and lipid metabolism in juvenile Chinese mitten crab, Eriocheir sinensis. One control diet (PL-devoid) and six test diets containing three PL types (soybean lecithin, egg yolk lecithin, and krill oil) at two levels (1% and 3%) each were formulated to feed E. sinensis juveniles (0.26 ± 0.01 g) for 8 weeks. Final body weight, specific growth rate, weight gain and molting frequency significantly increased in the PL supplementation groups, and the crabs fed 3% krill oil showed the best growth performance. The total lipid of whole body was significantly lower in the PL supplementation groups than that in the control. With PL supplementation, the specific activities of superoxide dismutase and catalase in the hepatopancreas markedly increased, while malondialdehyde decreased. In addition, total lipid and triglyceride contents in the hepatopancreas significantly decreased in the PL supplementation groups. Real-time quantitative PCR revealed that dietary PL reduced hepatic lipid accumulation probably through inhibiting lipid synthesis related genes and promoting lipid export related genes at the transcriptional level. This study indicates that krill oil is an excellent PL source and the diet with 3% PL can improve growth performance, increase antioxidant capacity and promote lipid utilization for juvenile E. sinensis.

A series of phenylaminopyrimidines has been identified as inhibitors of Janus kinases (JAKs). Development of this initial series led to the potent JAK2/JAK1 inhibitor CYT387 (N-(cyanomethyl)-4-[2-[[4-(4-morpholinyl)phenyl]amino]-4-pyrimidinyl]-benzamide). Details of synthesis and SAR studies of these compounds are reported.

7-Oxo-7H-naphtho[1,2,3-de]quinoline-11-carboxamides and analogues were prepared and evaluated for in vitro and in vivo antitumor activity. Chromophore variations included 'deaza' (7-oxo-7H-benz[de]anthracene) and 'diaza' (7-oxo-7H-benzo[e]perimidine) analogues, and side chain variations included chiral alpha-methyl compounds. The naphthoquinolines were the most cytotoxic, with IC(50) values of 5-20 nM, and showed the strongest DNA binding, with high selectivity for G-C rich DNA. The chiral alpha-methyl analogues were 10-20-fold more cytotoxic than the parent des-methyl compound. Both enantiomers provided substantial growth delays against s.c. colon 38 tumors in mice, with the R-enantiomer more active than the S (tumor growth delays of >35 and 12 days, respectively).

Pyrazolo[3,4,5- kl ]acridines were prepared by reaction of ethyl 1,9-dioxo-1,2,3,4,9,10-hexahydroacridine-4-carboxylate ( 4 ) with hydrazine and its methyl and 2-(dimethylamino)ethyl derivatives, followed by aromatization of the intermediate products with 1,4-benzoquinone. Conversion of the ester function to a carboxamide was also carried out and N -(2-(dimethylamino)ethyl)-1-(2-(dimethylamino)ethyl)-1,2-dihydropyrazolo[3,4,5- kl ]acridine-5-carboxamide ( 13c ) was appreciably cytotoxic in a panel of cell lines. Reaction of 4 with 4-methoxyphenylhydrazine gave instead a novel indolo[2,3- a ]acridine derivative.

A series of 7-oxo-7H-dibenz[f,ij]isoquinoline and 7-oxo-7H-benzo[e]perimidines bearing cationic side chains were prepared from aminoanthraquinones. The perimidines were prepared from 1-aminoanthraquinone by initial condensation with urea or dimethylacetamide. A series of 2-, 4-, 8-, and 11-carboxy derivatives of the dibenzisoquinolines were prepared from aminoanthraquinonecarboxylic acids. The cationic derivatives were prepared from these via amide, amine, or methylene linkers to study the effects of side chain positioning on biological activity. Within the series of carboxamide-linked compounds, the order of increasing cytotoxicity was 8- < 4- < 2- < 11-. The 2- and 4-carboxamides showed substantial growth delays against in vivo subcutaneous colon 38 tumors in mice, but the 11-carboxamide had curative activity in this refractory model and is being investigated further.

Cardiovascular disease is a major cause of mortality with the underlying process being atherosclerosis. Modified proteoglycans bind low-density lipoproteins (LDL), a critical initial step in the atherosclerotic cascade, representing a potential therapeutic target. Platelet-derived growth factor (PDGF) stimulates proteoglycan synthesis and is strongly implicated in atherogenesis. In human vascular smooth muscle cells (VSMCs), CYC10424 (Cytopia Research Ltd), a pyrido-pyrimidine derivative, dose-dependently decreased PDGF-mediated radiolabel incorporation into proteoglycans associated with an increase in electrophoretic mobility by SDS-PAGE. PDGF stimulated increases in both chemically-cleaved and xyloside-associated glycosaminoglycan (GAG) chain size, which were inhibited in the presence of CYC10424 by size exclusion chromatography (Sepharose CL-6B). CYC10424 treatment inhibited the PDGF effect to increase the 6:4 position sulfation ratio of monosulfated disaccharides by fluorophore-assisted carbohydrate electrophoresis. Proteoglycans derived from cells treated with CYC10424 had a decreased binding affinity and capacity to human LDL by gel mobility shift assay. CYC10424 and related compounds are possible candidates as therapeutic agents for the prevention of lipid deposition as characteristic of diseases such as atherosclerosis.

We describe the synthesis of a series of DNA-threading bis(9-aminoacridine-4-carboxamides) comprising ethylpiperidino and N -methylpiperidin-4-yl sidechains, joined via neutral flexible alkyl chains, charged flexible polyamine chains and a semi-rigid charged piperazine linker. Their cytotoxicity towards human leukaemic cells gives IC 50 values ranging from 99 to 1100 nM, with the ethylpiperidino series generally being more cytotoxic than the N -methylpiperidin-4-yl series. Measurements with supercoiled DNA indicate that they bisintercalate.

We have synthesized a series of bis(9-aminoacridine-4-carboxamides) linked via the 9-position with neutral flexible alkyl chains, charged flexible polyamine chains, and a semirigid charged piperazine-containing chain. The carboxamide side chains comprise N,N-dimethylaminoethyl and ethylmorpholino groups. The compounds are designed to bisintercalate into DNA by a threading mode, in which the side chains are intended to form hydrogen-bonding contacts with the O6/N7 atoms of guanine in the major groove, and the linkers are intended to lie in the minor groove. By this means, we anticipate that they will dissociate slowly from DNA, and be cytotoxic as a consequence of template inhibition of transcription. The dimers remove and reverse the supercoiling of closed circular DNA with helix unwinding angles ranging from 26 degrees to 46 degrees, confirming bifunctional intercalation in all cases, and the DNA complexes of representative members dissociate many orders of magnitude more slowly than simple aminoacridines. Cytotoxicity for human leukemic CCRF-CEM cells was determined, the most active agents having IC(50) values of 35-50 nM in a range extending over 20-fold, with neither the dimethylaminoethyl nor the ethylmorpholino series being intrinsically more toxic. In common with established transcription inhibitors, the morpholino series, with one exception, have no effect on cell cycle distribution in randomly dividing CCRF-CEM populations. By contrast, the dimethylaminoethyl series, with two exceptions, cause G2/M arrest in the manner of topoisomerase poisons, consistent with possible involvement of topoisomerases in their mode of action. Thus, the cellular response to these bisintercalating threading agents is complex and appears to be determined by both their side chain and linker structures. There are no simple relationships between structure, cytotoxicity, and cell cycle arrest, and the origins of this complexity are unclear given that the compounds bind to DNA by a common mechanism.

A series of 7-oxo-7H-dibenz[f,ij]isoquinoline and 7-oxo-7H-benzo[e]perimidines bearing cationic side chains were prepared from aminoanthraquinones. The perimidines were prepared from 1-aminoanthraquinone by initial condensation with urea or dimethylacetamide. A series of 2-, 4-, 8-, and 11-carboxy derivatives of the dibenzisoquinolines were prepared from aminoanthraquinonecarboxylic acids. The cationic derivatives were prepared from these via amide, amine, or methylene linkers to study the effects of side chain positioning on biological activity. Within the series of carboxamide-linked compounds, the order of increasing cytotoxicity was 8- < 4- < 2- < 11-. The 2- and 4-carboxamides showed substantial growth delays against in vivo subcutaneous colon 38 tumors in mice, but the 11-carboxamide had curative activity in this refractory model and is being investigated further.

Spinoflavanone B was prepared for the first time by the condensation of 2-hydroxy-4, 6-dimethoxy-methoxy-3,5-diprenylacetophenone with benzaldehyde followed by cyclization and demethoxymethylation.

The first total synthesis of two new natural prenylflavonoids, exiguaflavanone K (1) and (±)-leachianone G (2), has been achieved by condensation of 2-hydroxy-3-prenyl-4,6-dimethoxymethoxyacetophenone (5) with hydroxy-protected benzaldehydes 6 and 7, respectively, followed by cyclization and demethoxymethylation.

The acid precursors of the title compounds were prepared from methyl 2-amino-3-formylbenzoate (3), by FriedlAnder synthesis with o-methoxy- and o-nitro-phenylacetic acids, phenylpyruvic acid and benzo[b]thiophen-2-one, respectively. Except for the last example, cyclization of an initial 3-arylquinoline derivative was then required to give the tetracycle. Growth inhibition properties of the carboxamides in a series of cancer cell lines were measured for comparison with previous data for an isomeric series. In all cases, the present set were found to be less active.