Masao Hisadome

A novel anti-rheumatic drug suppresses tumor necrosis factor-α and augments interleukin-10 in adjuvant arthritic rats

Tumor necrosis factor-alpha (TNF-alpha) plays an important role in the pathology of rheumatoid arthritis. When N-[1-(4-¿[4-(pyrimidin-2-yl)piperazin-1-yl]methyl¿phenyl)cycloprop yl] acetamide (Y-39041) (3-30 mg/kg) was orally administered to rats with established arthritis from day 15 to day 20, hindpaw volume was significantly reduced. This inhibitory activity of Y-39041 was kept up after administration was stopped. On day 17 Y-39041 suppressed lipopolysaccharide-induced TNF-alpha and interleukin-6 production in serum at doses of 3-30 mg/kg, and augmented interleukin-10 production at doses of 10 and 30 mg/kg. The finding that Y-39041 suppresses TNF-alpha and interleukin-6 production and augments interleukin-10 production could be beneficial in the therapy of chronic inflammatory diseases.

EVALUATION OF COMBINED ANTIBIOTIC-OMEPRAZOLE THERAPIES IN HELICOBACTER PYLORI-INFECTED MONGOLIAN GERBILS

Abstract: Mongolian gerbils are a laboratory host for gastric colonization with Helicobacter pylori, showing gastritis followed by typical gastric ulcer after infection with H. pylori. In such gerbils, we evaluated combined therapies of amoxicillin (AMPC) and clarithromycin (CAM) as antibiotics, and omeprazole (OPZ) as a H+/K+ adenosine triphosphatase (ATPase) inhibitor. The gerbils were orally inoculated with 2 × 108 bacilli of H. pylori ATCC 43504. Four weeks after inoculation, the infected gerbils were orally treated singly with OPZ, AMPC, and CAM, and their insufficient efficacy on bacterial clearance was confirmed by a polymerase chain reaction technique, and by a culture method. In contrast, combined therapy of OPZ plus either AMPC or CAM showed significant bacterial clearance, demonstrating the efficacy of this combined therapy in the gerbil model. Mongolian gerbils are suggested to be useful for the pharmacological evaluation of anti-H. pylori compounds.

A novel dual regulator of tumor necrosis factor-α and interleukin-10 protects mice from endotoxin-induced shock

A pyrimidylpiperazine derivative, N-[1-(4-¿[4-(pyrimidin-2-yl)piperazin-1-yl]methyl¿phenyl)cycloprop yl] acetamide (Y-39041), is a dual cytokine regulator of tumor necrosis factor (TNF)-alpha and interleukin-10 production. Lipopolysaccharide-induced TNF-alpha release in BALB/c mice was inhibited by the oral treatment with the compound at 10-100 mg/kg (about 80% suppression) while interleukin-10 release was augmented (about 10-fold increase at 30 mg/kg). In addition, Y-39041 (30 mg/kg, p.o.) completely protected mice from lipopolysaccharide-induced death by the treatment before and after lipopolysaccharide injection. The finding that Y-39041 suppresses TNF-alpha production and stimulates interleukin-10 production at the same time provides new insights for the treatment of septic shock, rheumatoid arthritis and Crohns diseases.

Novel DMARDs on the basis of a new concept of dual cytokine regulation, TNF-α suppression and IL-10 augmentation

A series of arylpiperazine derivatives was synthesized to obtain agents showing apparent therapeutic effects in a chronic inflammatory animal model, starting from a lead possessing potent dual cytokine regulatory activity in vivo. We found a pyrimidylpiperazine derivative 17c showing the dual regulatory activity and an excellent therapeutic effect in an adjuvant-induced arthritis model.

Novel phenylpiperazine derivatives as dual cytokine regulators with TNF-α suppressing and IL-10 augmenting activity

Phenylpiperazine derivatives were synthesized as dual cytokine regulators with TNF-alpha suppressing and IL-10 augmenting activity. Lead optimization led to compound 5k having the potent regulatory activity and demonstrating remarkable protective effects against the lethal challenge of LPS in mice. suggesting that 5k would be a promising drug candidate for the treatment of TNF-alpha associated diseases including septic shock.

Treatment with Y‐40138, a multiple cytokine production modulator, inhibits lipopolysaccharide‐ or tumour necrosis factor‐α‐induced production of pro‐inflammatory cytokines and augments interleukin‐10

N‐[1‐(4‐[4‐(pyrimidin‐2‐yl)piperazin‐1‐yl]methyl phenyl)cyclopropyl] acetamide.HCl (Y‐40138) suppresses liver injury in concanavalin A‐ and d‐galactosamine/lipopolysaccharide (LPS)‐induced mouse hepatitis models. However, the mechanism of action of Y‐40138 has not been fully investigated. In this study, we examined the effect of Y‐40138 on cytokine production in mice. Cytokine production was induced by intraperitoneal injection of LPS (0.5 mg kg−1) or intravenous injection of recombinant mouse tumour necrosis factor (TNF)‐α (10μg mouse−1) in BALB/c mice. TNF‐α and interleukin (IL)‐10 reached maximum levels 1.5 h after the LPS injection. IL‐12 and interferon‐γ (IFN‐γ) reached maximum levels 3 to 9 h after the injection. When Y‐40138 was orally administered 30 min prior to the injection, it inhibited TNF‐α, IL‐12 and IFN‐γ production and augmented IL‐10 production. Y‐40138 also inhibited IL‐12 production and augmented IL‐10 production in TNF‐α‐stimulated mice. In IL‐10 knockout mice, Y‐40138 inhibited TNF‐α and IL‐12 production 1.5h after the LPS injection but not after 3 h or later, unlike in wild mice. In addition, TNF‐α production was inhibited by Y‐40138 at concentrations that could not augment IL‐10 production. These data suggest that Y‐40138 modulates pro‐inflammatory cytokine production by both IL‐10‐dependent and ‐independent mechanisms.

( + )-3-[4-(2-dimethylamino-1-methylethoxy)-phenyl]-1H-pyrazolo[3,4-b]pyridine-1-acetic acid (Y-25510) stimulates production of IL-1β and IL-6 at the level of messenger RNA expression in cultured human monocytes

(+/-)-3-[4-(2-Dimethylamino-1-methylethoxy)phenyl]-1H-pyrazolo[3, 4-b]pyridine-1-acetic acid (Y-25510) stimulated the mRNA expression for interleukin-1 beta (IL-1 beta), and enhanced the expression induced by lipopolysaccharide (LPS) in cultured human peripheral blood mononuclear cells (PBMC) and THP-1 cells, a cell-line derived from human monocytic leukemia. Y-25510 also stimulated the mRNA expression for IL-6 in both types of the cells, however, the stimulation required the presence of LPS. In THP-1 cells, the stimulation of IL-1 beta mRNA expression by Y-25510 was suppressed by cycloheximide, an inhibitor of protein synthesis. This phenomenon indicates that the stimulation requires de norv protein synthesis. In contrast, the stimulation of mRNA expression for IL-6 by Y-25510 was not suppressed by cycloheximide but suppressed by N alpha-p-tosyl-L-phenylalanine chloromethyl ketone (TPCK), an inhibitor of nuclear transcription factor-kappa B (NF-kappa B) activation, in the presence of LPS, suggesting that the stimulation requires NF-kappa activation. These results demonstrate that Y-25510 stimulates the mRNA expression for IL-1 beta and IL-6 by different mechanisms. Dexamethasone suppressed the LPS-induced expression of mRNA for IL-1 beta and IL-6 in THP-1 cells, whereas the drug never suppressed the mRNA expression for these cytokines in the presence of Y-25510. The result indicates that Y-25510 stimulates the mRNA expression for IL-1 beta and IL-6 by different mechanisms from those of LPS.

Enhancement of in vivo production of IL-1α and IL-6 in mice by Y-25510, a 1H-pyrazolo[3,4-b]pyridine-1-acetic acid derivative

The serum concentrations of interleukin(IL)-1 alpha and IL-6 in C57BL/6 and C3H/HeN mice reached the maximum at 12-16 h after the intravenous treatment with (+/-)-3-[4-(2-dimethylamino-1-methylethoxy)- phenyl]-1H-pyrazolo[3,4-b]pyridine-1-acetic acid (Y-25510) at a dose of 3 mg/kg, and the concentration of IL-10 did at 20 h after the treatment. By repeated treatments with Y-25510 to C57BL/6 mice for 14 days, the maximal values of IL-1 alpha and IL-6 at day 14 were respectively 6.6 times and 5.7 times relative to those on day 1. Neither the counts of peripheral leukocytes nor those of platelets were, however, increased until day 15. The repeated treatment with Y-25510 followed by anti-IL-10 antibody for 14 days was significantly more effective than that with Y-25510 alone in increasing the concentrations of IL-1 alpha and IL-6 in C3H/HeN mice. In addition, both the counts of peripheral leukocytes and platelets were significantly increased at day 18. In conclusion, Y-25510 enhanced not only the production of endogenous IL-1 alpha and IL-6 but also that of IL-10 in healthy mice. As a result, in normal conditions, both the counts of peripheral leukocytes and platelets were never increased because of the inhibitory effect of endogenously produced IL-10.

Enhancement of host defense by Y-25510, (±)-3-[4-(2-dimethylamino-1-methylethoxy)phenyl]-1H-pyrazolo[3,4-b]pyridine-1-acetic acid, a novel synthetic compound. A comparison with recombinant human granulocyte colony-stimulating factor in 5-fluorouracil-treated mice

The effect of a novel synthetic compound, Y-25510, (+-)-3-[4-(2-dimethylamino-1-methylethoxy)phenyl]-1H-pyrazolo[3,4 -b] pyridine-1-acetic acid, on recovery from long-lasting leukopenia induced by 5-fluorouracil was compared with that of recombinant human granulocyte colony-stimulating factor (rhG-CSF). When mice were administered i.p. with 5-FU (200 mg/kg) on days 0 and 7, intravenous administration of Y-25510 (100 and 1000 micrograms/kg) prevented the decrease in the peripheral leukocyte and neutrophil number and accelerated the recovery from leukopenia. Subcutaneous administration of rhG-CSF (50 micrograms/kg) did not prevent leukopenia but accelerated the recovery from leukopenia. In particular, peripheral neutrophil number increased over a normal level. The administration of Y-25510 (10, 100 and 1000 micrograms/kg) restored the decrease in the number of bone marrow cells, spleen cells, lymphocytes, neutrophils and monocytes. The administration of rhG-CSF (50 micrograms/kg) restored the decrease in the number of bone marrow cells, spleen cells, and neutrophils but not that of lymphocytes and monocytes. In fractions of bone marrow cells on day 21, the administration of Y-25510 (1000 micrograms/kg) showed a tendency of restoring the decrease in neutrophil number. In conclusion, the administration of Y-25510 prevented leukopenia and accelerated the recovery from leukopenia in the 5-FU-treated mice. It is suggested that the mechanism of the restorative action of Y-25510 is different from that of rhG-CSF. In a number of immature bone marrow cells Y-25510 has a potent stimulatory effect on the recovery from the decrease in number of hematopoietic cells, keeping a balance in number of each blood cell.