Grelan Pharmaceutical Co., Ltd.

Monoclonal antibodies (Mabs) are valuable reagents for the purification, characterization and immunolocalization of proteins. In this study, we raised Mabs against human peroxisome proliferator-activated receptors (PPARs) using baculovirus particles displaying surface glycoprotein gp64-fusion proteins as the immunizing agent. In this system, to display fusion proteins on the viral surface, the amino terminal sequences of human PPARd and PPARg2 are inserted in-frame between the signal sequence and the mature domain of the gp64 nucleotide sequence.Mabs were raised by immunization with whole virus without a purification of the target antigens. The Mabs generated by this novel method were shown to recognize not only the gp64-PPARs fusion protein, but also mature, expressed proteins by a wide variety of techniques, including immunohistochemistry, immunoblotting, and electrophoretic mobility shift assays (EMSAs). Transfection of the transfer vector containing a nucleotide sequence encoding less than 30 amino acids along with linearized baculovirus DNA allows for the production of a high affinity antibody against the corresponding mature form. This method is of potential utility in that it allows the production of valuable antibodies without the requirement of a protein purification step.

We investigated the antinociceptive effect of pentazocine hydrochloride (pentazocine) in combination with morphine hydrochloride (morphine) using two antinociceptive tests; i.e., the tail-immersion and scald-pain tests, in rats.In the tail-immersion test, the rat's tail was immersed in warm water at 47°, and the latency to a nociceptive response was measured.In the scald-pain test, the right hind foot was scalded by immersion into hot water at 57°.Two hours later, addnl. thermal stimulus was applied to the same foot, and the latency to a nociceptive response was measured.S.c. treatment with either pentazocine (6, 12, 24 mg/kg) or morphine (1.5, 3, 6 mg/kg) alone dose-dependently showed antinociceptive effects in both tests.The ED₅₀ values (95% confidence limit) of pentazocine and morphine were 13.0 (5.4-31.5) and 2.4 (1.6-3.7) mg/kg in the tail-immersion test and 11.0 (4.5-26.6) and 3.8 (1.8-7.2) mg/kg in the scald-pain test, resp.Simultaneous treatment with pentazocine at the similar dose augmented the morphine (1.5 mg/kg)-induced antinociception, but did not diminish the morphine (6 mg/kg)-induced antinociception in both tests.These results suggest that the simultaneous administration of pentazocine at the antinociceptive dose and morphine exerts addnl. antinociceptive activity against thermal and scald-induced inflammatory pain.

In this study, a very reliable HPLC method was developed for the determination of fenofibric acid and reduced fenofibric acid in the biological samples described as follows. After addition of the internal standard solution and 0.5 M HCl to the biological sample, fenofibric acid, reduced fenofibric acid and the internal standard were extracted with a mixed solvent of n-hexane and ethyl acetate (90:10) from the mixture. The acids were back-extracted from the organic phase with 0.1 M Na2HPO4 and then re-extracted from the aqueous phase with a mixed solution of n-hexane and ethyl acetate (95:5) after addition of 0.5 M HCl. The organic phase was evaporated to dryness under the vacuum. The residue was dissolved in MeOH and diluted with distilled water. An aliquot of the resulting solution was injected on the HPLC. High reproducibility was observed in this HPLC method (C.V.% less than 4%). Moreover it was confirmed that the conjugates in the urine could be hydrolyzed by incubation at 37 degrees C for 18 h after addition of 400 IU of beta-glucuronidase.