• 한국양식학회지
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• 제11권2호
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• pp.151-158
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• 1998

동자개의 배란유도에 대한 LHRH-a, pimozide 및 HCG의 복강내 투여효과에 대해 조사하였다. 어체중 kg당 HCG를 각각 5,000, 10,0000, 20,000 IU를 주사하였을 경우 71.4, 80, 100%의 배란유도율을 나타냈다. 또한 pimozide의 존재에 관계없이 LHRH-a를 주사한 개체들 대부분이 배란되었다. LHRH-a 단독투여 실험에서 kg당 200, 200, $400^{\mu}$g 처리에서 각각 50, 75 및 100%의 배란유도율을 나타냈다. pimozide첨가는LHRH-a의 작용을 향상시켰다. 즉, LHRH-a와 pimozide의 혼합투여에서 어체중 kg당 pimozide 1 mg과 LHRH-a $300^{\mu}$g에서 100%의 가장 좋은 배란유도율을 나타내었다. 그러나 pimozide 단독투여에서 배란이 유도되지 않았다. 호르몬 처리후의 생식소숙도지수(GSI)와 pesudo-GSI의 변화를 조사한 결과, HCG, LHRH-a 및 LHRH-a+pimozide를 주사한 그룹간의 생식소숙도지수(GSI)와 pseudo-GSI에 있어서 퉁계적인 유의차는 없었다(P>0.05). 호르모처리후 배란이 일어나기까지의 소요시간은 HCG처리후 17~22시간, LHRH-a단독 또는 pimozide와의 혼합처리에서 대체로 23~29시간이 걸렸다.

• 약학회지
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• 제38권2호
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• pp.202-210
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• 1994

To investigate the feasibility of mucosal delivery of $[D-Ala^6]$ LHRH, a potent analogue of LHRH, enzymatic proteolysis of $[D-Ala^6]$ LHRH and inhibitory effect of medium chain fatty acid salts(MFA) were studied using rabbit mucosal homogenate. $[D-Ala^6]$ LHRH incubated in homogenates of rectal(RE), nasal(NA) and vaginal(VA) mucosa were assayed by HPLC. The degradation of $[D-Ala^6]$ LHRH followed the first order kinetics. The degradation products were found as $[D-Ala^6]$ $LHRH^{1-7}$(m-i), to a lesser extent, $[D-Ala^6]$ $LHRH^{1-9}$(m-ii) and $[D-Ala^6]$ $LHRH^{1-3}$(m-iii) by the method of amino acid analysis(PITC method). The formation of$[D-Ala^6]$ $LHRH^{1-7}$ was not inhibited by the addition of disodium ethylenediaminetetraacetic acid but inhibited by sodium tauro-24,25-dihydrofusidate, suggesting that endopeptidase 24.11(EP 24.11) cleaves the $Leu^7-Arg^8$ bond of $[D-Ala^6]$ LHRH and is the primary $[D-Ala^6]$ LHRH degrading enzyme. The patterns of $[D-Ala^6]$ LHRH degradation indicated that EP 24.11 exists in each mucosal homogenate with the order of RE>NA>VA. MFA significantly inhibited the proteolysis of $[D-Ala^6]$ LHRH. The addition of sodium caprate(1.0%) or sodium laurate(0.5%) to the each mucosal homogenate completely protected $[D-Ala^6]$ LHRH from the degradation.

• 한국동물학회지
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• 제32권4호
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• pp.329-338
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• 1989

Present study examined the effect of intermittent versus continuous infusion of progesterone(P) on LHRH release in uiuo from the mediobasal hvpothalamus of ovariectomiEed, estradiol-primed adult rats bearing push-pull cannulae. Three experimental groups were studied: 1) control (perfusion medium only),2) intermittent perfusion of P (10-min on,20-min off, and 3) continuous perfusion of p. p (10 ng/mll was directly infused into the MBH following a 3 hr basal collection. Perfusates were collected at 10 min intents린s on ice and LHRH release was measured by LHRH radioimmunoassav. Cycle detector analysis revealed that the spontaneous HRH output in the control group was pulsatile over a 7 hr push-pull perfusion period. The mean basal LHRH release, pulse amplitude and pl서se period were 0.68 $\pm$ 0.03 ps110 min, 1.15 $\pm$0.08 pg and 60 $\pm$ 9 min, respectivelv. Intermi구eat perfusion of P clearly stimulated the mean LHRH release (pre-P vs post-P: 1.14 $\pm$ 0.18 vs 1.99 $\pm$ 0.53 pg) without changes in LHRH pulse frequency. In contrast to intermittent infusion of p, continuous administration of P faithed to modify LHRH release, since the mean LHRH release and pulse amplitude between pre-P and post-P perfusion urere similar. The in vitro study clearly showed that intermittent, but not continuous administration of P is effective in stimulating LHRH release. Therefore, it appears that rhythmic secretion of P mal be the erective signal for activating the neural LHRH apparatus.

• 한국동물학회지
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• 제32권3호
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• pp.275-280
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• 1989

본 연구는 한우에서 적출한 시상하부조직을 superfusion 하여 체외배양 하면서 in vitro LHRH의 내인성 분비양상을 조사하였으며, 한우의 in vitro LHRH 분비양상과 수입소인 홀스타인 젖소의 LHRH 분비양상을 비교하였다. 시상하부의 median eminence조직을 적출하여 조각을 내어서 superfusion chamber에 넣은 후 37$^{\circ}C$ 배양기에서 배양하였다. superfusion은 4시간동안 계속되었으며, 10분간격으로 분획을 받아내어 LHRH 분비능을 LHRH방사면역 측정법으로 측정하였다. LHRH 분비 양상의 특성은 PULSAR algorithm으로 분석하였다. 한우와 수입 젖소에 있어서 내인성 LHRH분비는 맥동적인 양상을 보였다. 한우에 있어서 LHRH 분비의 평균분비, 펄스의 진폭과 그 간격은 11.08 $\pm$ 1.50 pg/min/mg x 10-$^2$, 21.43 1 7.28 pg/mg x 10-$^2$, and 39.42 $\pm$ 3.08 min이었고 이 값은 홀스타인 젖소에서의 측정치와 거의 유사하였다. 이와 같은 한우의 펄스 발신기의 기본적인 특징은 한우의 신경내분비적 연구에 중요하리라 사료된다.

• Archives of Pharmacal Research
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• 제18권5호
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• pp.325-331
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• 1995

The objective of this study was to find a rational dosage form for vaginal mucosal delivery of LHRH. Vaginal absorption of LHRH was estimated by measuring its ovulation inducting effect in rat and in vitro vaginal membrane permeation study in rabbit. THe effects of different hydrogel bases, such as Polycarbophil and Pemulen compared with solutions on vaginal membrane permeation of LHRH were investigated. Sodium laurate, disodium ethylenediamine brane permeaiton of LHRH were investigated. Sodium ethylenediamine tetraacetate (EDTA) and sodium tauro-24, 25-dihydrofusidate (STDHF), which are effective peptidase inhibitors were chosen as additives to a LHRH hydrogel delivery system and LHRH solutions. A Polycarbophil compared with a solution formulation 3.4 times increase in LHRH vaginal membrane permeability compared with a solution formulation. Vaginal membrane permeability from the Polycarbophil was greater than that from Pemulen hydrogels. This may be due to the larger bioadhesive values. LHRH solution with EDTA(2%), STDHF(1%) and sodlaurate(0.5%) showed 4.1 times, 4.8 times and 6.0 times of ovulation inducing activity compared with control. These results suggest that enzyme inhibition effect of EDTA, STDHF and sod, laurate may be result in substantial enhancement of vaginal absorption. By administraiton of Polycarbophil hydrogels containing LHRH the ovulation inducing activity was 3.3 times greater than the solutions. This result indicates the bioadhesive hydrogels as well as peptidase in hibition significantly improved absorption of LHRH. By coadministration with these inhibitors the ovulation inducing activity of Polycarbophi hydrogel containing LHRH was comparable with subcutaneous administration in ovulation inducing activity.

• Bulletin of the Korean Chemical Society
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• 제32권11호
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• pp.4021-4026
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• 2011

Functions of the luteinizing hormone releasing hormone (LHRH) and its induced release by divalent metal ions have received great attention because this neurotransmitter subsequently regulates the secretion of luteinizing hormone (LH). Metal-LHRH complexes were synthesized by addition of various Cu(II),Ni(II),Zn(II) ions into LHRH in order to understand how the induced release of LHRH is possible. The degree of complexation was monitored by $^1H$, $^{13}C$-NMR chemical shifts, and final products were identified by Mass spectrometry. Solutionstate structure determination of Zn(II)-LHRH out of metal-complexes was accomplished by using NMR and NMR-based distance geometry (DG). Interproton distance information from nuclear Overhauser effect spectroscopy was utilized for structure determination. Structure obtained in this study has a cyclic conformation exhibiting a specific ${\alpha}$-helical turn with residue numbers His[2]-Leu[7] out of 10 amino acids. Comparison of chemical shifts and EPR studies of Ni(II),Cu(II)-LHRH complexes exhibit that these metal complexes have 4-coordination geometry.

• 약학회지
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• 제38권4호
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• pp.440-450
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• 1994

Due to the limited bioavailability of $[D-Ala^6]$LHRH from nonparenteral transmucosal sites of administration, enhancement of mucosal permeability by coadministration of several protease inhibitors and/or penetration enhancers were studied in rabbit mucosa. As a reliable bioassay method for $[D-Ala^6]$LHRH, ovulation-inducing effect were measured after vaginal administration in the rat. The permeation of $[D-Ala^6]$LHRH through the mucosal membrane of rabbit mounted on George-Grass diffusion cells were examined in the presence of polyoxyethylene 9-lauryl ether (POE), ${\beta}$-cyclodextrin$({\beta}-CyD)$ or ethylene diamine tetra acetate disodium salt(EDTA). The vaginal membrane showed higher permeability of $[D-Ala^6]$LHRH than the rectal and nasal membrane. POE and ${\beta}-CyD$ showed a small promoting effect on the membrane permeation of $[D-Ala^6]$LHRH, but EDTA showed significant enhancement. Ovaluation was enhanced by the coadministration of sodium laurate(0.5%), a protease inhibitor but was not enhanced by EDTA, a penetration enhancer. On the other hands, coadministration of sodium tauro 24,25 dihydrofusidate(1%) and EDTA(2%) enhanced the ovulation inducing-effect 2.8 times. These results suggest that the vaginal administration of $[D-Ala^6]$LHRH with STDHF or sodium laurate as a protease inhibitor, and EDTA as a penetration enhancer, may become an elective method for transmucosal delivery of $[D-Ala^6]$ LHRH.

• 한국자기공명학회논문지
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• 제13권2호
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• pp.143-153
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• 2009

Luteinizing Hormone Releasing Hormone (LHRH) is composed of 10 amino acids, and is best known as a neurotransmitter. Because of the 80% homology in animals, much more concerns have focused on the substances that have similar functions or can control LHRH. Ni, Cu-LHRH complexes were synthesized. The degree of complexation was monitored by $^1H,\;^{13}C$-NMR chemical shifts, and final products were identified by ESI-Mass spectrum. Solution-state structure determination of Ni-LHRH complex was accomplished by using NMR results and NMR-based distance geometry (DG). Interproton distances from nuclear Overhauser effect spectroscopy (NOESY) were utilized for the molecular structure determination. Results were compared with previous structures obtained from energy minimization and other spectroscopic methods. Structure obtained in this study has a cyclic conformation which is similar to that of energy minimized, and exhibits a specific a-helical turn with residue numbers (2~7) out of 10 amino acids. Comparison of chemical shifts and EPR studies of Ni, Cu-LHRH complexes exhibit that Ni-LHRH complex has same binding sites with the 4-coordination mode as in Zn-LHRH complex.

• Journal of Pharmaceutical Investigation
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• 제27권1호
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• pp.15-22
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• 1997

The mucosal route of administration(nasal, buccal, conjunctival and vaginal) has recently been considered as an alternative to parenteral delivery for many peptide drugs because enzymatic degradation of these agents may be partly avoided. The objective of these study was to establish the optimal mucosal administration dosage form of $LHRH/[D-Ala^6]LHRH$, based on presystemic metabolism. We reported previously the peptidase inhibition effect of medium chain fatty acid salts(sodium caprylate, soadium caprate and sodium laurate), EDTA and STDHF on the proteolysis of $LHRH/[D-Ala^6]LHRH$ in rabbit mucosal homgenates. We also reported that EDTA, STDHF and sodium laurate markedly increased the potency of $LHRH/[D-Ala^6]LHRH$ solution applied vaginally. In the present study, by administration of polycarbophil hydrogel containing LHRH the ovulation inducing activity was 3.3 times greater than solution. These results indicate not only peptidase inhibitor but also polycarbophil hydrogel significantly improved the absorption of this drug. The results of this study would provide the feasibility as a rational dosage form for improving bioavailability and self administration of this hydrogel by the vaginal application.

• Clinical and Experimental Reproductive Medicine
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• 제18권2호
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• pp.189-196
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• 1991

Long-term administration of luteinizing hormone-releasing hormone(LHRH) agonists, through a process of pituitary desensitization and down-regulation of receptors, inhibits the secretion of gonadotropin and sex-steroids and induces a reversible suppression of gonadal activity. This approach can be used as an effective endocrine therapy for some hormone-dependent tumors. We have used D-Trp6-LHRH, a long acting LHRH agonist, for the treatment of eleven patients with uterine leiomyomas, thereafter myomectomy was performed in seven cases and observed the ultrastructural changes of leiomyoma with an electron microscope. The use of LHRH agonist may be effective in reducing the size of a myoma considerably by primarily inducing medical hypophysectomy and would allow easier surgical removal. Electron microscopic findings of myoma cells after the use of LHRH agonist included the following: loss of cristae and swelling nuclear chromatin, perinuclear vacuolation in cytoplasm. Bone mineral density was slightly decreased, however, the difference was not statistically significant.