Dihydropyrazine Derivatives as a New Type of DNA Strand Breaking Agent.

Yamaguchi, Tadatoshi; Kashige, Nobuhiro; Mishiro, Noriko; Miake, Fumio; Watanabe, Kenji

doi:10.1248/bpb.19.1261

Cited by 37 publications

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“…It is already apparent 1) that Cu 2ϩ converted into Cu 1ϩ by the reducing ability of DHPs. The relationship between the reducing ability shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…1) employed were all synthesized by condensation of diketones and diamines. 2,3-Dihydro-5,6-dimethylpyrazine (DHP-1), 2,3-dihydro-2,5,6-trimethylpyrazine (DHP-2) and 3-hydro-2,2,5,6-tetramethylpyrazine (DHP-3) were synthesized by the method of Yamaguchi et al 1) 2,3,-Dihydro-5-methy, 6-phenylpyrazine 13) (DHP-4) was also synthesized by similar method.…”

Section: Synthesis Of Dihydropyrazine Derivativesmentioning

confidence: 99%

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Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

Kashige

Takeuchi

Matsumoto

et al. 2005

Biological & Pharmaceutical Bulletin

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Dihydropyrazines (DHPs) revealed the single strand-cleavage 1) of covalently closed circular DNA of plasmid pBR322, especially in the presence of Cu 2ϩ . This cleavage was determined to be initiated by the radical species generated from DHPs. Therefore, further investigation of the reactivity of DHPs continues our laboratory.DNA damage is closely related to the variety of biological phenomena such as mutagenesis, carcinogenesis, aging and radiation effects. A better understanding of the mechanism of the biological effects caused by DHPs is of great interest, because DHPs were derived from sugar and universally existed in human body, and consequently might caused various internal injuries in vivo.2) The chemical reactivity 3,4) and the biological effects 2,5) of DHPs have been elucidated. The electron spin resonance (ESR) spectrum of DHPs showed the cogeneration of four radical species 6) such as · OH, · OOH, · CHR 2 and · CR 3 at the same time. Furthermore, the specificity 7) of the nucleotide sequence DNA strand cleavage sites produced by DHPs has been identified. However, whether oxygen radica'ls or carbon-centered radicals (C-radicals) primarily involved in the DNA strand-cleavage remains unclear. There are a number of reports of DNA strand-cleavage due to reactive oxygen species, including hydroxyl radical, however, there have been few reports related to C-radical. [8][9][10][11] The DNA damage caused by the copper-peroxide complex provides a unique example. 12) Herein, we use the results of DNA strand-cleavage activity and the ESR spectra with a spin-trapping agent to propose that the radical species that attacked the DNA strand formed 8-hydroxydeoxyguanosine (8-OHdG). MATERIALS AND METHODS Synthesis of Dihydropyrazine DerivativesThe dihydropyrazine derivatives ( Fig. 1) employed were all synthesized by condensation of diketones and diamines. 2,3-Dihydro-5,6-dimethylpyrazine (DHP-1), 2,3-dihydro-2,5,6-trimethylpyrazine (DHP-2) and 3-hydro-2,2,5,6-tetramethylpyrazine (DHP-3) were synthesized by the method of Yamaguchi et al. 1) 2,3,-Dihydro-5-methy, 6-phenylpyrazine 13) (DHP-4) was also synthesized by similar method.Assay of DNA Strand-Breaking Activity The method of assaying the DNA strand-cleavage activity of DHPs, using a covalently closed circular duplex DNA of plasmid pBR32 (ccc-DNA) was previously described. 14)ESR Spectroscopy of Dihydropyrazines The ESR spectra were recorded on a JES-FA200 spectrometer (JEOL Co., Tokyo) using a Mn 2ϩ marker as an external standard, and an ES-LC12 flat cell (JEOL Co., Tokyo). The spectra were measured in a 50 mM Tris-HCl buffer (pH 7.1) using 5,5-dimethy-1-pyrroline N-oxide (DMPO) as a spin trapping agent, according to previous paper. 6) The relative values were summarized in Table 3. The instrumental condition were: field center 335.9 mT, scan width Ϯ5 mT, modulation frequency 100 kHz, modulation width 0.14 mT, time constant 0.3 s, amplitude 7ϫ100, microwave power 10 mW, and microwave frequency 9.427 GHz. The spectra were recorded at 30 min after mixing.The Measureme...

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“…It is already apparent 1) that Cu 2ϩ converted into Cu 1ϩ by the reducing ability of DHPs. The relationship between the reducing ability shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Synthesis Of Dihydropyrazine Derivativesmentioning

confidence: 99%

Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

Kashige

Takeuchi

Matsumoto

et al. 2005

Biological & Pharmaceutical Bulletin

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“…Entretanto, como reportado em diversos estudos sobre os produtos da condensação cíclica do ALA, esta não é a única forma pela qual duas moléculas de ALA podem condensar para produzir produtos cíclicos 190 . Foi observado que a dimerização cíclica do ALA a pH 5-6 gera exclusivamente um derivado de dihidropirazina, o ácido 3,6-dihidropirazina-2,5-dipropanóico (DHPY) (Esquema 3).Vários estudos mostram que alguns derivados de dihidropirazinas podem causar quebras de fita em DNA plasmidial 191 pela geração de radicais · OH e centrados no carbono 192,193 . Esses danos podem ser facilitados pela alta afinidade do cobre ao DNA 194 .…”

Section: Danos Ao Dna Promovidos Por Derivados De Dihidropirazinaunclassified

Danos ao DNA promovidos por ácido 5-aminolevulínico: possível associação com o desenvolvimento de carcinoma hepatocelular em portadores de porfiria aguda intermitente

Onuki¹,

Teixeira²,

Medeiros³

et al. 2002

Quím. Nova

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Recebido em 26/6/01; aceito em 1/10/01 DNA DAMAGE INDUCED BY 5-AMINOLEVULINIC ACID: A POSSIBLE ASSOCIATION WITH THE DEVELOPMENT OF HEPATOCELLULAR CARCINOMA IN ACUTE INTERMITTENT PORPHYRIA PATIENTS. 5-Aminolevulinic acid (ALA) is a heme precursor accumulated in acute intermittent porphyria (AIP), which might be associated with hepatocellular carcinoma (HCC) in symptomatic patients. Under metal catalyzed oxidation, ALA and its cyclic dimerization product, 3,6-dihydropyrazine-2,5-dipropanoic acid, produce reactive oxygen species that damage plasmid and calf thymus DNA bases, increase the steady state level of 8-oxo-7,8-dihydro-2´-deoxyguanosine in liver DNA and promote mitochondrial DNA damage. The final product of ALA, 4,5-dioxovaleric acid (DOVA), is able to alkylate guanine moieties, producing adducts. ALA and DOVA are mutagenic in bacteria. This review shows an up-to-date literature data that reinforce the hypothesis that the DNA damage induced by ALA may be associated with the development of HCC in AIP patients.Keywords: 5-aminolevulinic acid; acute intermitent porphyria; DNA damage. PORFIRIASAs porfirias são definidas como doenças inatas ou adquiridas, relacionadas à deficiência de enzimas -inibição ou insuficiência -da via biossintética do grupo heme (Esquema 1) 1 . Esta deficiência resulta na produção excessiva e excreção de porfirinas e de seus precursores 2-6 . As porfirias são tradicionalmente classificadas com base no principal órgão que produz os metabólitos porfíricos em excesso (fígado ou os tecidos eritropoiéticos), na ocorrência ou não de ataques agudos e nas lesões cutâneas (fotossensibilidade) e/ou neurológicas produzidas 5,7,8 . Tipo de porfiria Aguda Hepática CutâneaPorfiria deficiente em ALA desidratase X X Porfiria aguda intermitente (AIP) X X Coproporfiria hereditária (HCP) X X X Porfiria variegata (VP) X X X Porfiria cutânea tarda (PCT) X X Porfiria eritropoiética congênita (CEP) X Protoporfiria eritropoiética (EPP) X A natureza molecular do defeito genético é altamente heterogê-nea e defeitos múltiplos têm sido demonstrados em cada tipo de porfiria 3,5 . Alguns modelos animais para os estudos dos diferentes tipos de porfiria já existem naturalmente, no caso de EPP e CEP e alguns foram desenvolvidos em laboratório, para EPP e porfirias hepáticas 9,10

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“…The DHPs revealed single strand-cleavage 3) on the covalently closed circular DNA of plasmid pBR322, especially in the presence of Cu 2ϩ . Since the participation of Cu 2ϩ enhanced the cleaving activity, it is likely that the effects of DHPs on E. coli also would be influenced.…”

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The Role of Dihydropyrazines in Accelerated Death of Escherichia coli on Addition of Copper(II)

Takeda

Takechi

Katoh

et al. 2005

Biological & Pharmaceutical Bulletin

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In a recent paper, 2) we demonstrated the biological effects, such as growth inhibition and mutagenesis in Escherichia coli, of dihydropyrazines (DHPs). The DHPs revealed single strand-cleavage 3) on the covalently closed circular DNA of plasmid pBR322, especially in the presence of Cu 2ϩ . Since the participation of Cu 2ϩ enhanced the cleaving activity, it is likely that the effects of DHPs on E. coli also would be influenced. Here, we examined the effects of DHPs on E. coli upon the addition of Cu 2ϩ . Pyrazine derivatives have been detected in the urine of humans with diabetes mellitus, 4) which suggests that DHPs, which are precursors of pyrazines, might be involved in diabetes mellitus, although this remains unconfirmed. DHPs also are produced from the accumulation of the heme precursor, 5-aminolevulinic acid (ALA), in lead poisoning and some types of porphyries. 5,6) We have reported that DHPs induce apoptosis of cultured cells, 7) and, therefore investigated other potential biological effects caused by DHPs. The sugar origin of DHPs and their prevalence in the human body suggests that these compounds may be involved in various internal injuries in vivo. 7) MATERIALS AND METHODS Synthesis of DHPsThe DHPs (Fig. 1) were synthesized by condensation of diketones and diamines. 2,3-Dihydro-5,6-dimethylpyrazine (DHP-1), 2,3-dihydro-2,5,6-trimethylpyrazine (DHP-2), and 3-hydro-2,2,5,6-tetramethylpyrazine (DHP-3) were prepared by the method of Yamaguchi et al. 3)Bacterial Strains E. coli K-12 strains used include the following. AB1157 [ara-14 argE3 galK2 his-4 lacY1 leuB6 mtl-1 proA2 rpsL31 supE44 thi- Survival Assay E. coli cultured overnight were diluted 100-fold in fresh LB medium and grown at 37°C until the OD 600 was approximately 0.1. Cells were challenged with DHPs for 1 h, incubated with shaking at 37°C, plated on LB agar, and incubated overnight at 37°C. Survival rates were calculated as the percentage of cells surviving after challenge by DHP-1, DHP-2 or DHP-3. All experiments were conducted at least four times.Enzymes and Chemicals Bovine erythrocyte superoxide dismutase (5000 units/mg), bovine liver catalase (11500 units/mg), dimethyl sulfoxide (DMSO), and 2-mercaptoethylamine hydrochloride (MEA) were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). RESULTSThe effect of DHPs in the presence of Cu 2ϩ was examined through the measurement of the E. coli viability. A wild-type strain, recA, and uvrB strain were incubated with or without Cu 2ϩ using three DHP types as shown in Fig. 1, which revealed in vitro DNA strand-cleaving activity of the order DHP-3ϾDHP-2ϾDHP-1.3) As shown in Fig. 2, the effect of DHPs in all strains of E. coli was accelerated greatly by addition of Cu 2ϩ . This is definitive evidence that the recA and uvrB strains possess greater sensitivity to DHPs upon addition of Cu 2ϩ compared to the wild-type strain. The effect in the recA strain was obvious, even without Cu 2ϩ . Therefore, this effect on E. coli must involve specific DNA damage caused by radical species generat...

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Dihydropyrazine Derivatives as a New Type of DNA Strand Breaking Agent.

Cited by 37 publications

References 0 publications

Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

Radical Species in DNA Strand-Cleavage Caused by Dihydropyrazines

Danos ao DNA promovidos por ácido 5-aminolevulínico: possível associação com o desenvolvimento de carcinoma hepatocelular em portadores de porfiria aguda intermitente

The Role of Dihydropyrazines in Accelerated Death of Escherichia coli on Addition of Copper(II)

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