• Biomolecules & Therapeutics
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• v.3 no.3
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• pp.233-237
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• 1995

The growth effects of newly developed recombinant human growth hormone (rHGH), were compared with those of Biotropine. For the effective evaluation, we examined the increasing rate of body weight and the thickness of tibial epiphysis as criteria of growth effects on hypophysectomised female rats treated with varing concentration of rHGH for 4 days. rHGH treated groups showed significant body weight gain which was less evident in Biotropine and vehicle treatment group. In tibial epiphyseal test, rHGH also showed clear effects compared to Biotropine and vehicle treatment group. Above findings indicate that newly developed rHGH has better effects of growth stimulation on female rats than Biotropine does.

• Biomolecules & Therapeutics
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• v.3 no.4
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• pp.251-255
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• 1995

DA-3002 is a genuine human growth hormone produced by Dong-A Pharm. Co. Ltd. research laboratory using recombinant DNA technic. In this study, antigenic potential of DA-3002 was examined by active systemic anaphyaxis(ASA) in guinea pigs, mouse-rat passive cutaneous anaphylaxis(PCA) and passive hemagglutination(PHA) test as a part of safety research. DA-3002 induced anaphylactic shock in ASA test using guinea pigs Immunized with DA-3002 alone or DA-3002 incoporated into Freund's complete adjutant(FCA) when challenged with 10 times higher dose of anticipated clinical dose of DA-3002. In the mouse-rat PCA and PHA test, DA-3002 also showed positive results. DA-3002, therfore, was considered to produce IgE, IgG, and/or IgM in mice. The results of this study were similar to those of the other human growth hormones and these positive results were thought to be caused due to the fact that both DA-3002 and the other human growth hormones were heterogenous proteins to guinea pigs and mice. Considering the fact that DA-3002 is a genuine human growth hormone of which structure is identical with indigenous human growth hormone, DA-3002 is thought not to cause immunological problems in clinical use.

• BMB Reports
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• v.28 no.5
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• pp.437-442
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• 1995

The potency of yeast-derived methionyl-free human growth hormone (rhGH), which was obtained by removal of the N-terminal Met from methionyl-hGH, was estimated by in vitro and in vivo assays. In radio-receptor assay where the binding affinity of growth hormone to the receptor was estimated, the recombinant hGH showed 2.9 international units (IU) per mg of specific activity. In contrast, pitUitary-derived human growth hormone had a slightly lower receptor binding activity (2.5 IU/mg) compared with recombinant growth hormone. For the in vivo assay, efficacy of rhGH was tested by use of hypophysectomized rats, in which pituitary organs were surgically removed, resulting in the termination of growth hormone secretion. The weight-increase in rats by the injection of rhGH was almost identical to the result obtained by the injection of the same amount of pituitary-derived (international standard) hGH. A comparision of the secondary structures of rhGH and rMet-hGH by circular dichroism spectrophotometer demonstrated that the removal of the methionyl residue from rMet-hGH did not exert any effect on the structure of the growth hormone. In conclusion, methionyl-free human growth hormone produced from yeast was highly potent in biological activity and maintained a legitimate three dimensional structure.

• Biomolecules & Therapeutics
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• v.3 no.1
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• pp.63-71
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• 1995

The local irritation studies of DA-3002, an authentic recombinant human growth hormone (rhGH), were carried out in rabbits after the following treatment ; application into the conjunctival sac of the eye (single), single subcutaneous and intramuscular injection, 7-day repeated subcutaneous and intramuscular injection. The results obtained were as follows. In the result of ocular irritation test, 0.16% solution of DA-3002 could be considered as a non-irritating material. In single subcutaneous and intramuscular irritation test, the irritancy of 0.16% DA-3002 solution was not so much different from that of saline. The local irritation of DA-3002 by 7-day repeated injection was negligible and similar to that of saline by both subcutaneous and intramuscular routes. These results suggest that DA-3002 has no irritating activity when injected through subcutaneous or intramuscular route for clinical practice as 0.16% solution.

• Toxicological Research
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• v.9 no.1
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• pp.125-132
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• 1993

Antigenic potential of recombinant human growth hormone (LBD-007), a newly developed drug for growth hormone deficiency, was investigated in mice and guinea pigs. 1. Mice showed production of antibodies against LBD-009 (1.5IU/kg) with aluminum hydroxide gel(alum) as an adjuvant, Judaged by the heterologous anaphylaxis (PCA) test using rats. On the other hand, antibodies against ovalbumin (OVA) inoculated with alum were definitely detected. 2. In the studies with guinea pigs, both the inoculation of LBD-009 (0.15IU/kg-1.5IU/kg) only and of LBD-009 with complete Freund's adjuvant (CFA) as an adjuvant did produce weak positive reactions in homologous passive cutaneous anaphylaxis (PCA). On the other hand, the inoculation of ovalbumin with complete Freund's adjuvant (CFA) produced positive reaction in PCA.

• Proceedings of the PSK Conference
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• 2002.04a
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• pp.127.2-128
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• 2002

• Biomolecules & Therapeutics
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• v.2 no.2
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• pp.161-172
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• 1994

This study was conducted to examine DA-3002, a biosynthetic human growth hormone, for its acute and subacute toxicities in mice and rats. The drug was administered subcutaneously and orally at a dose level of 1.0, 3.0, 8.9, 26.7 or 80.0 lU/kg once for single dose toxicity and given subcutaneously at a dose level of 0.34, 1.7 or 8.4 lU/kg daily for 13 weeks to investigate repeated dose toxicity. In the acute toxicity study, doses up to 80 lU/kg had no adverse effect on the behavior or body weight gain. Pathological examinations revealed no abnormal changes which could be attributed to toxic effect of DA-3002. In the subacute toxicity study, the growth hormone was tolerated well in broth mice and rats. No drug related deaths occurred and all animals appeared to be normal throughout the dosing period. Increases in body weight gain, food utilisation and absolute organ weights were observed in the rats in the high dose group. Mild changes in the blood chemical parameters were also seen in the treated groups. Histopathologically, however, no abnormal changes were observed in any organ. The changes noted during the treatment periods presumably represent exaggerated pharmacological effects of the growth hormone, and no observed adverse effect level (NOAEL) was considered to be more than 8.4 lu/kg/day.

• Proceedings of the Korean Society of Toxicology Conference
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• 1997.10a
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• pp.38-42
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• 1997

Human growth hormone is known as one of the peptide hormones which is consisted of 191 amino acids derived from the pituitary gland in humans. The objectives of this study were to supply inexpensive recombinant methionyl human growth hormones (rHGH) synthesized by the DNA technology in a yeast cell line and followed by the establishement of protein purification techniques. The next steps of the research were to study its physic-chemical properties and biological properties, and to evaluate various preclinical aspcts including pharmacokinetics sutdy, general pharmacology study, general toxicity test, and specific toxicity tests. Clinical phase I, II, III studies were also done against growth hormone dficient children to reveal that growth promoting effects were similar compared with the natural HGH extracted from pituitary glands and commercially available rHGHs. The results could be summarized that (I) this yeast dervied rHGH have had excellent physico-chemical and biological properties in comparison with a natural HGH and other synthesized rHGHs, (2) we could not see any toxic side effects when very high doses were administered to the experimental animals, and (3) this growth hormone showed effectiveness in the growth stimulating to growth hormone deficient patients.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.554-556
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• 2003

This research was focused on expression, purification and characterization of ubiquitin-specific protease 1 (UBP1) expressed in recombinant Escherichia coli. Various systems were constructed by fusing polycationic fusion tails or fusion partners to the C- or N-terminus of the product protein. In particular, UBP1 containing 6 histidine residues at the N-terminal end showed best results in terms of expression level and purification efficiency. The N-terminal $6{\times}His-tagged$ UBP1 was overproduced in recombinant E. coli using high cell density cultivation technology and purified using immobilized metal affinity chromatography. The molecular weight of UBP1 was found to be 83,500 daltons. The optimum temperature and pH for the enzyme reaction when ubiquitin-human growth hormone (hGH) was used as a substrate were $40^{\circ}C$ and pH 8.0, respectively.

• YAKHAK HOEJI
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• v.34 no.6
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• pp.439-446
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• 1990

The general pharmacological actions of recombinant human growth hormone (rHGH) were investigated. It had hypothermic action but neither sedative nor analgesic action. No pharmacological effects were observed in isolated guinea pig ileum and tracheal muscle and rat fundus and uterus. Slight hypotensive action with no effect on respiration was revealed at a dose of 20 IU/kg i.v. of rHGH in rabbits. The rHGH exhibited a weak inhibitory action of glucose tolerance in normal rats, significantly lowered the blood glucose contents in adrenalectomized rats 20 min after i.v. administration (80IU/kg), and produced a significant inhibitory effect on in vitro glycerol release in epinephrine-stimulated epididymal fat pad segments of rats.