• Molecules and Cells
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• 제37권2호
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• pp.140-148
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• 2014

We identified four basic amino acid residues as nuclear localization signals (NLS) in the C-terminal domain of the prototype foamy viral (PFV) integrase (IN) protein that were essential for viral replication. We constructed seven point mutants in the C-terminal domain by changing the lysine and arginine at residues 305, 308, 313, 315, 318, 324, and 329 to threonine or proline, respectively, to identify residues conferring NLS activity. Our results showed that mutation of these residues had no effect on expression assembly, release of viral particles, or in vitro recombinant IN enzymatic activity. However, mutations at residues 305 (R ${\rightarrow}$ T), 313(R ${\rightarrow}$ T), 315(R ${\rightarrow}$ P), and 329(R ${\rightarrow}$ T) lead to the production of defective viral particles with loss of infectivity, whereas non-defective mutations at residues 308(R ${\rightarrow}$ T), 318(K ${\rightarrow}$ T), and 324(K ${\rightarrow}$ T) did not show any adverse effects on subsequent production or release of viral particles. Sub-cellular fractionation and immunostaining for viral protein PFV-IN and PFV-Gag localization revealed predominant cytoplasmic localization of PFV-IN in defective mutants, whereas cytoplasmic and nuclear localization of PFV-IN was observed in wild type and non-defective mutants. However sub-cellular localization of PFV-Gag resulted in predominant nuclear localization and less presence in the cytoplasm of the wild type and non-defective mutants. But defective mutants showed only nuclear localization of Gag. Therefore, we postulate that four basic arginine residues at 305, 313, 315 and 329 confer the karyoplilic properties of PFV-IN and are essential for successful viral integration and replication.

• Bulletin of the Korean Chemical Society
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• 제28권9호
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• pp.1510-1514
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• 2007

Recently, it has been reported that the inhibition of the strand transfer function of HIV-1 integrase is necessary to obtain significant antiviral activity. Accordingly, several compounds typified by aryl 1,3-diketo acids that can inhibit strand transfer reaction of HIV-1 IN have been identified. In this work, we synthesized new 4- hydroxy-5-azacoumarin-3-carbox(thio)amides (1a-h) and evaluated for the inhibition of HIV-1 IN strand transfer reaction with a brief SAR. Among synthesized, compound 1e was the most potent HIV-1 IN inhibitor with equipotent activity to that of L-708,906. Therefore, the 4-hydroxy-5-azacoumarin ring can be considered as a new scaffold in designing more potent of HIV-1 IN inhibitors for treatment of AIDS.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.163.2-163.2
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• 2000

• 약학회지
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• 제59권3호
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• pp.85-91
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• 2015

Highly active antiretroviral therapy (HAART) has reduced AIDS-related morbidity and mortality; however, it has been associated with metabolic abnormalities including dyslipidemia and dysglycemia depending on the regimens used. The aims of this study were to analyze the prescription patterns of antiretroviral agents and to examine the prevalence of lipid abnormalities among the prescriptions of HAART. The electronic medical records (EMR) of HIV patients were retrospectively reviewed from January 2007 to September 2012 based on our inclusion criteria. The patients who had taken HAART for at least 3 months were included in this study. The lipid profiles of patients on antiretrovirals (ARTs) were collected from his or her laboratory data, and dyslipidemia was defined as total cholesterol (TC) ${\geq}240mg/dL$ and triglycerides (TG) >200 mg/dL. Eighty-four prescriptions were discovered during the study period. Twenty-three prescriptions were the combination of two nucleoside reverse transcriptase inhibitors (NRTIs) and one non-nucleoside reverse transcriptase inhibitor (NNRTI). Fifty-three prescriptions were the combination of two NRTIs and one protease inhibitor (PI) and thirty-nine prescriptions of them included a PI booster. Eight prescriptions were the combination of two NRTIs and one integrase inhibitor. The Incidence of hypertriglyceridemia among the patients receiving HAART was totally about 41.7% (2NRTIs+PI regimen vs. 2NRTIs+NNRTI regimen vs. 2 NRTIs+integrase inhibitor regimen, 52% vs. 12.5% vs. 25%), but there was no incidence of hypercholesterolemia. This study investigated that the prescription medication patterns and dyslipidemia associated with lipid abnormalities among HIV patients receiving HAART. The types of HAART prescription regimens had an effect on the occurrence of hypertriglycemia. Further studies related to metabolic abnormalities and adverse effects of HIV patients on ARTs are needed in the near future.

• Journal of Microbiology and Biotechnology
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• 제5권3호
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• pp.132-137
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• 1995

The lambda integrase (lnt) is believed to bind to several arm and core sites of attP DNA in order to facilitate intasome formation. We have done systematic mutagenic analysis on all 5 arm sites and found that P1 is absolutely required for integration while P2 is not. We also found that all 3 P' arm sites(P'1, P'2, and P'3) are required for efficient integrative recombination. P'1, which is an important binding site for excision, also seems to be crucial for integration when preincubation of attP DNA with Int and IHF is performed before recombination. Preincubation assay revealed that preincubation with Int and IHF improved the efficiency of recombination of wild type attP DNA and demolished recombinations of P'1 mutant attP DNAs.

• 약학회지
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• 제53권2호
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• pp.89-92
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• 2009

Dicaffeoyltartaric acid has a structural feature consisting of two caffeic acid units separated by tartaric acid linker and has been found to be a potent inhibitor of HIV-1 integrase and an antioxidant. In this study, bis(pyronyl)acrylic acid esters joined through a 5-membered ring as a linker were synthesized as the analogues of dicaffeoyltartaric acid.

• Plant Resources
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• 제4권3호
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• pp.192-195
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• 2001

Human immunodeficiency virus (HIV), the causative agent of AIDS, still continues to spread rapidly in the world population, especially in Africa and Southeast Asia. At present, two kinds of therapeutic approaches are used for treatment of AIDS. One is to target HIV reverse transcriptase, which is responsible for the viral genome transcription. The other is to inhibit HIV pretense PR, which is essential for the processing of viral proteins. Drug combinations based on these approaches can reduce the blood virus to an undetectable level. However, a small amount of virus may lurk inside the immune cells in a dormant state. Another major obstacle of long-term treatment of the disease is remarkable mutation in HIV. Most of the clinical chemotherapeutic agents have one or more of these problems. High cost and harmful side-effects further reduced the desirability of these drugs. In the course our studies on development of anti-HIV agents from natural products, we investigated various crude drugs for their inhibitory activity against HIV-induced cytopathic effects (CPE) in culture cells, HIV-pretense (PR), HIV-reverse transcriptase (RT) including ribonuclease H (RNase H), and HIV integrase (INT). In the present paper, some inhibitory substances relating to the development of anti-HIV agents are reported.

• 대한바이러스학회지
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• 제26권1호
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• pp.107-114
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• 1996

본 연구는 인간면역결핍바이러스의 입자를 비이온성 계면활성제로 처리할 때 바이러스 입자구조에서 분리되어 방출되는 바이러스 구조단백질들의 분포를 sucrose gradient로 분석하여, 바이러스 입자를 구성하는 바이러스 구조단백질과 바이러스입자의 생물리학적 특성을 연구하였다. 바이러스입자들을 0.16% NP40 (Nonidet P-40)으로 처리할 때, 바이러스 capsid 단백질과 바이러스 막 단백질 (membrance protein)들은 다른 바이러스 구성성분들과 잘 분리되었다. 계면활성제처리에서 방출되지 않은 구성 성분들은 matrix 단백질, nucleocapsid 단백질, reverse transcriptase, integrase 및 바이러스 RNA genome로써, 이들은 subviral 구조를 형성한다. 이러한 결과는 상대적으로 다른 바이러스들의 capsid 단백질과 면역 결핍 바이러스의 capsid 단백질 (p24)를 비교할 때, 면역결핍바이러스의 capsid 단백질은 바이러스핵을 형성할 때, capsid 단백질 사이의 결합력이 매우 약한 것으로 추정된다. 또한 바이러스 조절단백질의 하나인 vpr 단백질을 함유하는 바이러스입자를 NP40 처리하여 분석하였을 때, vpr 단백질은 subviral 구조에 존재하는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제18권12호
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• pp.1953-1957
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• 2008

Recombinant Lactobacillus strains have been constructed using gene splicing by overlap extension (SOE). Primers were designed of which one end of an amplified product contained complementary sequences for an end of other amplified fragment. For efficient matching, we used an asymmetric PCR step that was effective at generating an excess of strands that would anneal in the final PCR. CP12, a recombinant fragment consisting of the integrase gene and attachment site of the bacteriophage A2, was constructed and inserted into the genome of Lactobacillus casei ATCC 393, yielding Lb. casei ATCC 393::XCP12. Another recombinant Lb. casei strain was constructed, where the egfp gene was a part of the construction. The EGFP produced from Lb. casei ATCC 393::XCEGFP14 was detected by Western blot hybridization. This simple and widely applicable approach has significant advantages over standard recombinant DNA techniques for Lactobacillus species.

• Bulletin of the Korean Chemical Society
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• 제32권1호
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• pp.208-212
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• 2011

In this work, the calculated nuclear quadrupole coupling constants of $^{17}O$ in some styrylquinoline conformers were presented. The calculations were carried out to find the relationships between the charge distribution of styrylquinolines and their pharmaceutical behavior and to explore the differences among the electronic structures of some conformers of these potent HIV IN inhibitors. Furthermore, the HIV IN inhibitory of R1 and R2 rotamers was compared. On the basis of our results: - Charge density on oxygen atoms of carboxyl moiety has a dominant role in the drug activity. - The a conformer in which a divalent hydrogen atom is a link, has more capability in antiviral drug treatment. - The R1 conformer, as a $Mg^{+2}$ chelating agent, is better than R2 conformer and thus it is more inhibitor of HIV IN.