The excretion mechanism of fibronectin (FN)-related substances into the urine of normal and passive Hey-mann nephritis (PHN) rats was studied using enzyme immunoassay and immunoblot analysis. In normal rats, a small amount (0.20±0.67 μg/d) of FN-related substances, composed of 55- and 65-kDa FN fragments derived from the central cell-binding (Cell) domain of FN, were constitutively excreted into the urine. When PHN was induced in rats by the injection of an anti-Fx1A antibody, an increased excretion (4.96±3.51 μg/d) of intact FN and large (Mr>100-kDa) FN fragments containing the Cell and the other functional domains were seen. The PHN induction also caused the appearance of a considerable amount of Cell domain-containing FN fragments in plasma. Both the renal cortex homogenates of normal and PHN rats were capable of degrading plasma FN to generated the Cell domain-containing large FN fragments. Degradation of FN by the renal cortex homogenate was shown to be due to metal and/or thiol proteinase(s). These results suggest that the PHN-induced urinary excretion of FN fragments may be due to the degradation of plasma FN by renal proteinases that may be leaked upon PHN induction.
We examined the effects of in vitro low-dose irradiation on myeloid leukemia cells (M1 cells) and found that it enhanced the colony-forming ability (CFA) of M1 cells in semi-solid agar. This enhancement was inhibited by treatment with a protein synthesis inhibitor, cycloheximide and with an RNA synthesis inhibitor, actinomycin D, after irradiation. These findings suggested that low-dose irradiation induced the synthesis of some proteins which were attributed to the enhancement of CFA. Since we expected that one species of these proteins were heat shock proteins (hsps), we attempted to detect the hsp70 family by the Western blotting method and inducible hsp70 mRNA by the RT-PCR method. Low-dose irradiation induced the expression of hsp70 mRNA, whereas the enhancement of hsp70 (an inducible isoform) and hsc70 (a constitutively expressed isoform) expression could not be found. Furthermore, the M1 cells showed thermoresistance 1h after low-dose pre-irradiation, and also showed radioresistance 4h after irradiation. This time difference after pre-irradiation might be attributed to the different species of proteins in showing resistance to lethal stress. Therefore, some proteins other than hsp70 were believed to be concerned with the augmentation of CFA and the induction of thermo- and radio-resistance.
The effect of regucalcin, a Ca2+-binding protein, on Ca2+/calmodulin-dependent phosphatase activity in rat renal cortex cytosol was investigated. The addition of Ca2+/calmodulin in the enzyme reaction mixture caused a significant increase in the dephosphorylation of p-nitrophenylphosphate and phosphotyrosine used as the substrate for phosphatase in rat renal cortex cytosol. The presence of regucalcin (10-6M) in the enzyme reaction mixture caused a complete inhibition of Ca2+/calmodulin-dependent phosphatase activity in renal cortex cytosol. A half miximum effect of regucalcin inhibition was seen at 10-8M concentration. Moreover, phosphatase activity of purified calcineurin was significantly enhanced by the addition of Ca2+/calmodilin. This enhancement was completely inhibited by the presence of regucalcin (10-7M). The inhibitory effect of regucalcin was not weakened by increasing concentrations of CaCl2 (10-6 to 10-4M). The present results suggest that regucalcin can inhibit Ca2+/calmodilin-dependent phosphatase activity in rat renal cortex.
As described previously (Mizushina Y., Tanaka N., Yagi H., Kurosawa T., Onoue M., Seto H., Horie T., Aoyagi N., Yamaoka M., Matsukage A., Yoshida S., and Sakaguchi K., Biochim. Biophys. Acta, 1308, 256-262, 1996), linoleic acid (LA) inhibits the activities of mammalian DNA polymerases. We found a natural product from a basidiomycete, Ganoderma lucidum, that enhances this effect of LA in a special manner. The structure was identified to be an ergosterol peroxide, 5, 8-epidioxy-5α, 8α-ergosta-6, 22E-dien-3β-ol by spectroscopic analyses. The ergosterol peroxide (EPO) itself scarcely inhibited the activities of calf thymus DNA polymerase α(pol. α) or rat DNA polymerase β(pol. β). However, when EPO at 0.25 mM was present, 10 μM or less of LA almost completely inhibited the pol. β activity, while almost complete inhibition by LA itself was acheived at 80 μM or higher. Interestingly, under the same conditions, EPO did not affect the LA-effect on pol. α. The action mode of the EPO was discussed.
Effects of a newly synthesized antiulcer agent, YJA20379-4, on gastric proton pump (H+/K+-ATPase)activity, Helicobacter pylori (H. pylori) growth, gastric acid secretion, and gastro-duodenal lesions, were examined in comparison with those of omeprazole. YJA20379-4 markedly inhibited the H+/K+-ATPase activity in a concentration-dependent manner and the inhibitory effect was invreased under a weak acidic condition; the IC50 values were 32 and 81 μM at pH 6.4 and 7.4, respectively. The inhibition was completely antagonized by 0.5 mM dithiothretiol (DTT). In addition, YJA20379-4 showed a significant anti-H. pylori activity determined by the agar dilution method. The value of minimum inhibitory concentration (MIC, 3.9-11.7 μg/ml) was at least 3 times more potent than that of omeprazole. In pylorus ligated rats, YJA20379-4 inhibited basal gastric acid secretion when administered by the intraduodenal route (ED50 : 23.6 mg/kg). In experimental ulcer models, YJA20379-4 administered by the oral route dose-dependently prevented the development of gastro-duodenal lesions in rats. Moreover, repeated administration of YJA20379-4 promoted the healing of gastric ulcers induced by acetic acid. On the basis of the data obtained, it is suggested that YJA20379-4 has a wide spectrum of antiulcer activities, and its mode of antiulcer actions is dependent on the inhibition of H+/K+-ATPase activity and H. pylori growth and the enhancement of a mucosal defense. Thus, YJA20379-4 might prove to be a beneficial therapy for gastritis and peptic ulcer diseases.
T-3762, a newly developed fluoroquinolone antimicrobial agent, ciprofloxacin (CPFX) and ofloxacin (OFLX) were administered intravenously to anesthetized dogs in intravenous infusion, and blood pressure, heart rate and plasma histamine concentrations were monitored. T-3762 decreased blood pressure by 12.0%, without alterations in heart rate and plasma histamine concentration, only when infused at 150 mg/min. CPFX and OFLX both produced a rapid decrease in blood pressure in a dose-related manner, with an accompanying decrease in heart rate, but to a lesser extent. After infusion at 150 mg/min, CPFX caused death in 2 animals within a few minutes, while OFLX produced maximum decreases in blood pressure and heart rate, by 69.0% abd 26.4%, respectively. The infusion of these 2 agents resulted in dose-related increases in plasma histamine concentrations parallel to the decreases in blood pressure : the miximum, attained with CPFX at 50 mg/min and OFLX at 150 mg/min, were 379.2 and 167.8 ng/ml, respectively. For CPFX and OFLX, the relationship between the maximum levels of decreased blood pressure and increased histamine concentration in plasma was highly significant. The hypotension induced by CPFX was efficiently reduced by the pretreatment of animals with antihistamines. The results from this study suggest that hypotension induced in dogs following the intravenous infusion of fluoroquinolone antimicrobial agents may be dependent on their ability to cause histamine release from cells and tissues, and indicates that T-3762 is devoid of this ability in comparison to CPFX and OFLX.
To predict the actions of T-3762, a newly developed fluoroquinolone antimicrobial agent, as well as ciprofloxacin (CPFX) and ofloxacin (OFLX), on injection sites when dosed parenterally, their ability to increase cutaneous vascular permeability in dogs and to release histamine from rat peritoneal mast cells was examined. CPFX and OFLX increased cutaneous vascular permeability in concentrations ranging from 16 to 32 μg/ml, while T-3762 was inactive at 2000 μg/ml. The vascular permeability-increasing activities of these drugs were inhibited efficiently by pretreatment with a combined dose of diphenhydramine and cimetidine. CPFX induced histamine release from rat mast cells in a dose-dependent manner, whereas T-3762 was ineffective. Therefore, it is concluded that fluoroquinolone antimicrobial agents may have the ability to cause an increase in cutaneous vascular permeability by releasing histamine from mast cells at the injection site when administered parenterally, and that T-3762 has minimum activity among the agents tested in this study.
Teleocidin derivatives and the core structure, (-)-indolactam-V ((-)-IL-V), adopt two conformations in solution, the "twist" and he "sofa" forms. (-)-Benzolactam-V8-310 ((-)-BL-V8-310), which specifically adopts the twist form in solution, has been reported to have a significant effect on HL-60 cells and protein kinase C affinity. In this paper, we describe the biological activity with regard to tumor promotion on mouse skin and the wide variety of biological activity of (-)-BL-V8-310 and its derivatives. In both twist and sofa forms (-)-BL-V8-310 inhibited specific 3H-12-O-tetradecanoylphorbol-13-acetate (TPA) binding to a particulate fraction of mouse skin more strongly than (-)-IL-V. The doses for 50% inhibition (IC50) of (-)-IL-V, (-)-BL-V8-310, and teleocidin B-4 were 1000, 400 and 12nM, respectively. As for the induction of tumor necrosis factor-α(TNF-α) release into the medium from HL-60 cells, the EC200 values, which are the concentrations of the compound required to achieve 200 pg/ml TNF-α in the medium, were 1700, 500 and 19 nM for (-)-IL-V, (-)-BL-V8-310 and telecidin B-4, respectively. The same amounts (5.5 nmol per application) of (-)-BL-V8-310 and teleocidin B-4, induced tumors on mouse skin initiated with 7, 12-dimethylbenz(a)anthracene (DMBA) in 13.3% abd 86.7% of tumor-bearing mice, respectively, in week 20. These results confirmed that the twist form of teleocidin derivatives is the active form as far as the induction of biological activity is concerned. Also (-)-BL-V8-310 is a new synthetic tumor promoter designed from data obtained using the receptor cavity model of TPA-type tumor promoters.
The antihypertensive effect of sesamin, a lignan from sesame oil, was examined using salt-loaded and unloaded stroke-prone spontaneously hypertensive rats (SHRSP). The animals at 6 weeks of age were separated into a salt-loaded group and an unloaded group. Salt-loaded animals were maintained on 1% NaCl drinking water. Each group was further divided into two groups : normal-diet group and sesamin-diet group. Systolic blood pressure of all animals was monitored once weekly. At the end of the feeding periods, cardiovascular hypertrophy and renal damage were evaluated. In the salt-loaded group, sesamin feeding significantly suppressed the development of hypertension, and efficient suppression was maintained from 9 to 26 weeks (e.g., 215±4 vs. 180±4mmHg, at 17 weeks old). The left ventricle plus septum weight-to-body weigth ratio was slightly but significantly lowered by sesamin feeding. When the degree of vascular hypertrophy of the aorta and superior mesenteric artery was histochemically evaluated, wall thickness and wall area of these vessels were significantly decreased by the sesamin feeding. Histological renal damage such as thickening of the tunica intima and fibrinoid degeneration of the arterial wall were often observed in the normal-diet group, but this damage was efficiently reduced in the sesamin-fed animal. On the other hand, in the salt-unloaded group, only a slight and nonsignificant suppressive effect of sesamin on th development of hypertension was observed. Although the wall area of the aorta was significantly decreased by the sesamin feeding, other vascular parameters were not ameliorated. The incidence of histological renal damage tended to decrease in sesamin-fed animals, but these alterations were not statistically significant. Thus, sesamin feeding was much more effective as an antihypertensive regimen in salt-loaded SHRSP than in unloaded SHRSP, thereby suggesting that sesamin is more useful as a prophylactic treatment in the malignant status of hypertension and/or hypertension followed by water ans salt retention.
The phylogenetic relationship of Atractylodes lancea, A. chinensis, A. koreana, A. ovata and A. japonica were analyzed by comparing the 2.6 kb sequence in a chloroplast gene trnK encoding tRNALys (UUU). The dried rhizomes of the former three species have been used as the crude drug "So-jutsu" and those of the latter two as "Byaku-jutsu" in Chinese and Japanese traditional medicine ("Kampo-medicine"). The trnK phylogenetic tree revealed that A. ovata is an outgroup of the five Atractylodes species examined and that A. japonica and A. lancea are most closely related. PCR amplification of trnK with HinfI digestion provided us with a simple method to distinguish A. ovata from other Atractylodes species at the molecular level.
Two strains of autotetraploid plants of Atractylodes lancea DC. (Compositae) were raised from the in vitro colchicine-treated shoot cultures, and field trials were performed to evaluate their growth and the amount of essential oil components in the rhizome in comparison with the corresponding diploids. The tetraploid plants had larger leaves than the diploids. One of the selected tetraploid lines had about 1.5 times as heavy rhizomes as the diploid and contained atractylodin, hinesol and β-eudesmol in the rhizome to as great an extent or slightly less than the diploids. However, the contents of these constituents in the rhizome of the other tetraploid strain were lower than the diploids. The chloroplast number per guard cell, stomatal length, and stomatal density of leaf lower epidermis of the shoot cultures were good indicators for distinguishing tetraploids from diploids.
The skin penetrative action of high purity cis-ω-12-octadecenoic acid (petroselinic acid, HP-PSA) on rat skin was compared with that of high purity cis-ω-9-octadecenoic acid (oleic acid, HP-OA), following treatment of rat intact skin surface with either 0.05M HP-PSA or HP-OA in propylene glycol (PG), using Fourier transform/attenuated total reflection (FT-IR/ATR) analysis. Both HP-PSA and HP-OA disordered the lipid structures of the stratum corneum region to a similar extent. Removal of the extractable lipids of the stratum corneum region was marked with HP-PSA/PG but was very slight upon HP-OA/PG treatment. The spectra of the amide II region which originated from proteins suggests that HP-PSA/PG more rapidly disordered the protein structures of both the stratum corneum and the dermis than HP-OA/PG. However, the extent of disordering of the protein structures was presumed to be similar between these two skin penetration enhancers at the maximum level. Enhancement of PG flux in the dermis showed strong positive correlation with the degree of dermisdisordering action of HP-PSA/PG and HP-OA/PG. These results demonstrate that HP-PSA, which has a double bond at an even numbered position (ω-12), more rapidly affects the perturbation of the structures of both the stratum corneum and the dermis than HP-OA, which has the double bond at an odd numvered position (ω-9). Differences in the physicochemical properties of HP-PSA and HP-OA which originate from differences in the double bond position most likely determine the efficacy of these compounds as skin penetration enhancers.
In order to explore the relationship between the pharmacokinetic properties and pharmacological actions of lipophilic drugs injected with lipid carrier systems, probucol was selected as a model drug with high lipophilicity, and the effect of disposition control on cholesterol-lowering activities was evaluated. Both large emulsion, with mean diameter of 280 nm, and long-circulating type small emulsion containing egg sphingomyelin with mean diameter of 100 nm, showed stable incorporation of probucol. The former produced rapid accumulation of producol in the liver, while the latter demonstrated prolonged systemic circulation and gradual hepatic uptake. On the other hand, injection of a micellar solution with HCO-60 (polyoxyethylene hydrogenated castor oil) showed a rapid decrease in plasma concentration and a high hepatic uptake of producol, similar to injections with serum, suggesting the rapid release of the drug from the micelles. However, probucol in a micellar solution showed higher cholesterol-lowering action than that in emulsion formulations. These results suggested that the pharmacological action of probucol in the liver might be affected by the uptake mode and sequential disposition in the organ, depending on the drug retention properties of the lipid carrier particles.
We investigated the antiviral mechanisms of K-252a, a broad non-speciflic protein kinase inhibitor which was isolated from Nocardiopsis sp. and its derivative (KT5926), against vesicular stomatitis virus (VSV) replication in BHK-21 cells. Although K-252a (5 μM) and KT5926 (15 μM) similarly suppressed the viral primary and secondary transcriptions and genomic RNA synthesis in vivo, the inhibitory mechanisms did not seem to be the same; phosphorylation of the viral NS protein was suppressed by K-252a, which might account for the decreased viral RNA synthesis caused by K-252a. On the other hand, KT5926, being known to preferentially inhibit myosin light chain kinase (MLCK), had little effect on NS protein phosphorylation. Cellular casein kinaes II, which is delieved to be involved in the phosphorylation of the N-terminal side (domain I) of NS protein, was not inhibited at all by KT5926 even at 15 μM under in vitro assay conditions, and was only weakly inhibited by K-252a at 1 to 10 μM. Neither inhibitor seemed to directly affect viral protein synthesis, but affected it indirectly as a secondary effect of reduced viral RNA synthesis. These results suggest that both the KT5926-sensitive and the KT5926-resistant but K-252a-sensitive functions are involved in the essential processes of viral RNA synthesis. The KT5926-sensitive function(s) might not be involved in the NS protein phosphorylation, but may participate in some other way in the process of virus replication. On the other hand, the KT5926-resistant, K-252a-sensitive function(s) are probably involved in NS protein phosphorylation. The possible nature of those functions is discussed.
Human saliva contains a proline-rich polypeptide, salivary peptide P-C, which potentiates insulin release and reduces glucagon release from perfused rat pancreas to decrease blood glucose level. To elucidate the process of secretion into humoral fluid of this peptide morphologically, we investigated ultrastructural localization of P-C in human submandibular gland by immunogold technique with anti-peptide P-C whose specificity to P-C was confirmed by immunoblotting. The labeling with gold particles which represents the distribution of P-C-like-immunoreactivity (P-C-LI) was detected in the secretory granules and rough endoplasmic reticula of the acinar serous cells and in few mucosa cells. P-C-LI was also observed in the lumen of striated duct but not intracellularly in the ductal cells themselves, indicating the P-C is not probably reabsorbed there. These results suggest that salivary peptide P-C is present in acinar serous cells, is secreted into the oral cavity, and may be reabsorbed through the digestive tract to modulate the blood glucose level after feeding.
A fluorescent derivative of GM1 [20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol], a main Ginseng saponin metabolite formed by intestinal bacteria, was obtained from the condensation of its trisnor-aldehyde derivative with dansyl hydrazine. The dansylated GM1 fluoresced strongly and showed almost the same properties as its parent compound in lipophilicity and biological activities, so this fluorescent compound might provide an insight into the mechanism of pharmacological activities of GM1.
Astilbin, a dihydroflavonol rhamnoside isolated from the leaves of Engelhardtia chrysolepis, enhanced the vanadate-stimulated release of lipoprotein lipase (LPL) activity from rat isolated fat pads. N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide (H-8), a potent inhibitor of cAMP-dependent protein kinase (PKA), markedly inhibited the enhancement by astilbin. Lipolysis in the fat pads was stimulated by astilbin alone in a dose-dependent manner and this stimulation was suppressed in the presence of vanadate, probably due to its antilipolytic action. A significant enhancement by astilbin was observed with increasing effects of vanadate on cAMP content in the fat pads and on cAMP phosphodiesterase (PDE) activity in the particulate fraction although astilbin alone showed only a slight increase in the cellular cAMP content and PDE activity. Astilbin may enhance the vanadate-stimulated release of LPL activity throuth a synergistic effect on an increase in the cellular cAMP content produced by vanadate accompanied by more potent activation of PKA.
The protective effect of ceftriaxone on isepamicin-induced nephorotoxicity was investigated. For 14d, Wistar rats were administered either ceftriaxone 100 mg/kg intraperitoneally, isepamicin 300 mg/kg subcutaneously, or ceftriaxone isepamicin in combination. The animals given 300 mg/kg of isepamicin showed a significant increase in urine NAG (N-acetyl-β-D-glucosaminidase) levels as compared with the control animals which received saline (p<0.01). However, the increase in NAG level was markedly less when isepamicin was administered in combination with ceftriaxone (p<0.01). Ceftriaxone alone had no effect on urine NAG activity. Serum creatinine levels were significantly higher in animals treated with isepamicin alone than in control animals (p<0.01) or animals receiving the isepamicin ceftriaxone combination (p<0.01). After 14 d of treatment, ceftriaxone had not accumulated in renal tissue, but it did reduce the renal intracortical accumulation of isepamicin (p<0.01). Histopathologically, ceftriaxone induced very few cellular alterations and considerably reduced the manifestation of typical signs of isepamicin nephrotoxicity. This investigation demonstrates that ceftriaxone protects animals against isepamicin-induced nephrotoxicity.
Change in cellular reduced glutathione (GSH) level was examined after the addition of 1-10 mM potassium sorbate (SA-K) to cultured rat hepatocytes. The cellular GSH content was decreased to the lowest level at 6 h after the addition of SA-K, and then gradually returned to the normal level except for hepatoxytes exposed to 10 mM SA-K. Although the decrease in GSH level was not associated with lactate dehydrogenase (LDH) leakage in hepatoxytes exposed to SA-K up to the concentration of 5 mM, cell injury was caused in cells exposed to 10 mM SA-K. When eicosapentaenoic acid was added in conjunction with various concentrations of SA-K to hepatocytes, peroxidation of the fatty acid was accelerated in parallel with the decrease in cellular GSH level. The enhanced lipid peroxidation in the hepatoxytes co-exposed to SA-K and eicosapentaenoic acid (EPA) induced the development of cell injury. These results suggest that hepatocytes exposed to SA-K become susceptible to oxidative stress such as lipid peroxidation.
The renal handling of doxorubicin (DXR) was investigated using a kidney epithelial cell line, LLC-PK1. The uptake of DXR by LLC-PK1 cells cultured on plastic dishes was shown to be temperature and concentration dependent. The initial uptake of DXR was slightly saturable. The Km and Vmax of the saturable component were calculated to be 20.2 μM, and 0.355 nmol/mg protein/10 min, respectively. The release of DXR from LLC-PK1 cells was very slow at 37°C and almost negligible at 4°C, indicating that most of the DXR in the cells irreversibly binds to cellular constituents and that only a slight amount of unbound DXR participates in the efflux out of the cells. DXR uptake at 37°C was significantly decreased in the presence of 2, 4-dinitrophenol. However, organic cations and aminoglycoside antibiotics, such as tetraethylammonium, N1-methylnicotinamide, guanidine, gentamicin and neomycin, did not inhibit DXR uptake, suggesting that a process distinct from the organic cation transport system and absorptive endocytosis might be involved in the uptake of DXR by LLC-PK1 cells.
i-OHAP, a major metabolite of aprindine (AP), was isolated by TLC from rat feces and identified as N-[3-(N, N-diethylamino)propyl]-N-phenyl-2-aminoindan-5-ol, based on 1H-NMR, the H-H correlation spectroscopy (COSY) spectrum, MS and LC-MS. Its structure was also confirmed by comparison with the synthesized compound. The hydroxy group of i-OHAP was located at the 5-position of the indan ring. AP is a prochiral compound, and the metabolism of AP to i-OHAP was stereoselective. The ratio of (+)/(-)-i-OHAP in rat feces and in human urine was about 5 and 15, respectively.
A convenient high-performance liquid chromatography (HPLC) was developed for the rapid assay of granisetron (GRN) in biological fluids, such as serum, urine, and pleural effusion, from cancer patients. Extrelut<[○!R]>-1 was used for the solid-phase extraction. HPLC was carried out using a LiChroCART<[○!R]> cartridge column packed with Lichrospher 100 CN and a mobile phase consisting of 0.1 M acetate buffer (pH 3.5) and acetonitrile (7 : 3). A fluorescence detector of 290 nm for excitation and 365 nm for emission was used. The standard curve was linear over the range of 2 to 100 ng/ml of GRN. Assay precision, expressed as a coefficient of variation (C.V.), was in the range of 0.9-5.4% in the within-day assay and 2.5-6.9% in the between-day assay, respectively. GRN was well separated on the HPLC chromatogram from drugs such as etoposide, metclopramide, ondansetron, and domperidone which are often used together with GRN. It was suggested that the present method is useful for the rapid monitoring of GRN in the serum, urine, and pleural effusion of patients undergoing cancer chemotherapy.
Intraperitoneal administration of 2, 5-dimethylpyrazine (2, 5-DMP) was found to inhibit oxytocin- and prostaglandin F2α -induced tetanic uterine contractions in normal or pregnant female rats. This suggests that 2, 5-DMP may be used as a countercontraction agent or relaxant for preventing oxytocic agent-induced medical accident including uterine rupture or pressure death of the fetus due to uterine contractions.
We studied the inhibition of S-warfarin metabolism by nonsteroidal antiinflammatory drugs (NSAIDs) in human liver microsomes in vitro. After screening for potential inhibitors among ten NSAIDs using human recombinant cytochrome P450, inhibition kinetic parameters were estimated using human liver microsomes. Phenylbutazone and bucolome were suggested to increase the unbound steady-state level of S-warfarin about four- and five-fold, respectively, as estimated from these metabolic parameters.
4-[N-(5, 6, 7, 8-Tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthyl)amino]benzoic acid (5) exhibited weak retinoidal and retinoid synergistic activities in HL-60 cell differentiation assay. N-Alkylation of 5 caused decrease or loss or differentiation-inducing activity, but enhanced the synergistic activity with a synthetic retinoid Am80 (2), as reflected in the potent synergistic EC50 (SEC50) values of DA023 (11, 1.6×10-10 M) and DA113 (14, 1.4×10-10 M) in the presence of 1.0×10-10 M Am80 (2).
Several arylmethylidene thiaxolidinediones were synthesized and their retinoidal activities were examined. TZ181 (7a), having a benzanilide skeleton, exhibited differentiation-inducing activity in HL-60 cell assay, while TZ191 (7b), the N-methylated analog of TZ181 (7a), TZ245 (9) and TZ335 (10) acted as retinoid synergists like the RXR-selective ligand, LGD1069 (5).