Small structural changes of the imidazopyridine diacylglycerol acyltransferase 2 (DGAT2) inhibitors produce an improved safety profile.

Small molecule DGAT2 inhibitors have shown promise for the treatment of metabolic diseases in preclinical models. Herein, we report the first toxicological evaluation of imidazopyridine-based DGAT2 inhibitors and show that the arteriopathy associated with imidazopyridine 1 can be mitigated with small structural modifications, and is thus not mechanism related.

[A comparative study of the effect of granisetron and a combination of granisetron plus steroids on cancer chemotherapy induced emesis].

Effects of 40 micrograms/kg of granisetron monotherapy (K group) and concurrent therapy with a steroid (KS group) on acute and delayed emesis induced by cancer chemotherapy which included CDDP at a dose of 60 mg/m2 or more were compared in random clinical trials under the central registration method. In KS group, either 500 mg of methylprednisolone succinate or 8 mg of dexamethasone phosphate was given prior to granisetron administration. Clinical symptoms such as vomiting, nausea and anorexia were better in KS group than in K group, on any day from day 1 to day 7, and there was a statistically significant difference on day 1 and day 2. The cumulative total control rate throughout the period of seven days was also significantly higher in KS group. KS group was rated higher in the final clinical evaluation based on doctor's impressions, but there was no significant difference between the two groups. Augmented antiemetic effect of granisetron by concurrent therapy with a steroid was most notably demonstrated in male patients under 60 years of age. The antiemetic effect at the acute stage was proven to influence the final clinical effectiveness, thus suggesting the importance of antiemetic therapy of acute emesis. Adverse reactions were seen in two out of 122 patients (1.6%). They were slight headache and moderate diarrhea in 1 case each, both of which disappeared soon, confirming the high safety profile of granisetron.

Preparation of monoclonal antibodies against the IL-6 signal transducer, gp130, that can inhibit IL-6-mediated functions.

mAbs specific to human gp130, a signal transducing component of the IL-6 receptor complex, were prepared by immunizing mice with a previously described recombinant human soluble gp130. Some of the mAbs inhibited the IL-6-induced association of soluble gp130 and soluble IL-6 receptor. Three mAbs (GPX7, GPX22 and GPZ35) were shown to inhibit IL-6-mediated biological responses such as Ig production in a human B cell line and proliferative responses of a human Lennert's lymphoma-derived T cell line, a human myeloma cell line, and a mouse pro-B cell line-derived transfectant expressing human gp130.

Association of recombinant soluble IL-6-signal transducer, gp130, with a complex of IL 6 and soluble IL-6 receptor, and establishment of an ELISA for soluble gp130.

IL-6 mediates its pleiotropic functions through two membrane proteins, a ligand-binding molecule (IL-6 receptor, IL-6R) and a non-ligand-binding signal transducer (gp130). Starting with a previously isolated cDNA clone encoding human gp130, recombinant soluble gp130 (sgp130) lacking the transmembrane and cytoplasmic regions was expressed in COS7 cells or CHO cells. sgp130 could associate with a complex of IL-6 and soluble IL-6R (sIL-6R), also lacking transmembrane and cytoplasmic regions. This indicated that extracellular region of gp130 was responsible for the association with IL-6R which was occupied by IL-6. An enzyme-linked immunosorbent assay (ELISA) for the quantitation of sgp130 was established, which was based on the interaction of sgp130 with the complex of IL-6 and sIL-6R and could detect sgp130 as low as 1 ng/ml.

Preparation of soluble murine IL-6 receptor and anti-murine IL-6 receptor antibodies.

Starting with a previously isolated cDNA clone encoding murine IL-6R, a stable transformed Chinese hamster ovary cell line constitutively expressing soluble murine IL-6R (smIL-6R) has been established. The smIL-6R was purified to homogeneity by sequential filtration and chromatography of culture medium. The smIL-6R augmented the sensitivity of M1 cells to IL-6 in their growth inhibition in a dose-response manner. Rat hybridomas producing mAb specific to murine IL-6R were also established. One of the clones, RS13, produced IgG2a isotype that was capable of inhibiting IL-6 activity. ELISA for the quantitation of smIL-6R was established, which could detect smIL-6R in a quantity as low as 1 ng/ml.

Site-specific mutagenesis of human interleukin-6 and its biological activity.

Amino acid substitutions of human interleukin-6 (IL-6) were performed. Single substitution Met162----Ala and double substitutions Leu159.166----Val resulted in a significant decrease of IL-6 activity in the production of immunoglobulin (Ig) from B-cells. Single substitution Leu166----Val or Leu159----Val gave a slight or no significant decrease in the Ig-induction activity, respectively. The receptor-binding activity of each IL-6 mutant was also examined. It was observed that the decrease of the receptor-binding activity was generally in parallel with that of the Ig-induction activity. We therefore suggest that hydrophobic side-chains existing in Met162, Leu159, and Leu166 are significantly involved in the receptor-binding of IL-6.

Chemical modification and 1H-NMR studies on the receptor-binding region of human interleukin 6.

Oxidation of the Met residues of human interleukin 6 (IL-6) molecule has been performed. Reactivity of Met for the oxidation reaction was found to decrease in the order of Met50, Met118, Met185, Met162, and Met68. Chemical modifications involving oxidation and carboxypeptidase A digestion of IL-6 have led to the assignments of the methyl proton resonances of Met162 and Met185, respectively. The hydroxynitrobenzyl chromophore attached to Trp158 in the IL-6 molecule showed a different absorption spectrum when the labeled IL-6 was bound to the soluble IL-6 receptor. This result indicates that Trp158 is near the receptor-binding region in IL-6. On the basis of the 1H-NMR and chemical modification data, it has been concluded that Trp158 is in spatial proximity to Met162, His165 and Met185. The receptor-binding activity decreased with an increase in the number of oxidized Met residues. Of these five Met residues, Met162 was the residue in which the receptor-binding activity decreased in the most parallel degree with that of the oxidation reaction.

[A case of hepatocellular carcinoma (HCC) with lung metastasis which responded to chemotherapy with a single use of 1-hexylcarbamoyl-5-fluorouracil (HCFU)].

To our knowledge, there has been only one report pertaining to the efficacy of HCFU for treatment of metastatic lung lesion of HCC, so we reported a case of HCC with lung metastasis which responded to chemotherapy with a single use of HCFU. A 64-year-old male was diagnosed as having HCC with lung metastasis by biochemical examination, abdominal CT, hepatic arteriogram and chest X-P. He had been treated previously with gamma-interferon and mitoxantrone, which were assessed as NC and PD, respectively. Two months after last chemotherapy, HCFU was administrated at a dose of 400 mg/body everyday for 8 months. After 4 weeks metastatic lung lesions showed remarkable regression (47% decrease) and disappeared completely 9 weeks later. The size of primary liver tumor gradually decreased during therapy and revealed marked improvement (85.7% decrease) after about 2 months of this therapy. During these periods serum levels of alpha-fetoprotein dropped from 430 ng/ml to less than 10 ng/ml. He is presently still alive and the duration of the PR attained to 35 weeks. As side effects, hypoproteinemia, anorexia and hot sensation were observed.

[Antitumoral effect of gamma-interferon in patients with hepatocellular carcinoma].

The antitumoral effect of gamma-Interferon (Re-IFN-gamma, KW-2202) on nine patients with hepatocellular carcinoma (HCC) has been investigated. gamma-IFN was administered intravenously biweekly at a dose of 8-24 x 10(6) units/day for 5 consecutive days. As all patients had measurable disease determined by an abdominal CT, the antitumoral effect was evaluated by CT, according to the criteria of Koyama and Saito, During gamma-IFN therapy, one patient, who received a total of 4.4 x 10(8) units of gamma-IFN, achieved a partial response (PR) 133 days after onset of treatment. Another patient showed a minor response (MR) 43 days after start of therapy. The duration of the PR and MR were 7.8 weeks and 10.8 weeks, respectively. Two patients were assessed as having had no change (NC), and 5 patients as still manifesting a progressive disease (PD). Marked falls in serum alpha-fetoprotein levels during therapy were observed in 2 cases of which one was graded as having obtained a PR, and the other, a NC. The toxicities observed were fever, general malaise, headaches, and joint pains, which were slight and transient in most cases. In one case, however, a therapy was stopped because of a continuous and severe shoulder pain with no metastasis. Further studies on the use of IFN, possibly in combination with other chemotherapeutic agents, should be performed in patients with HCC.