124-20-9Relevant academic research and scientific papers
Probing mammalian spermine oxidase enzyme-substrate complex through molecular modeling, site-directed mutagenesis and biochemical characterization
Tavladoraki, Paraskevi,Cervelli, Manuela,Antonangeli, Fabrizio,Minervini, Giovanni,Stano, Pasquale,Federico, Rodolfo,Mariottini, Paolo,Polticelli, Fabio
, p. 1115 - 1126 (2011)
Spermine oxidase (SMO) and acetylpolyamine oxidase (APAO) are FAD-dependent enzymes that are involved in the highly regulated pathways of polyamine biosynthesis and degradation. Polyamine content is strictly related to cell growth, and dysfunctions in polyamine metabolism have been linked with cancer. Specific inhibitors of SMO and APAO would allow analyzing the precise role of these enzymes in polyamine metabolism and related pathologies. However, none of the available polyamine oxidase inhibitors displays the desired characteristics of selective affinity and specificity. In addition, repeated efforts to obtain structural details at the atomic level on these two enzymes have all failed. In the present study, in an effort to better understand structure-function relationships, SMO enzyme-substrate complex has been probed through a combination of molecular modeling, site-directed mutagenesis and biochemical studies. Results obtained indicate that SMO binds spermine in a similar conformation as that observed in the yeast polyamine oxidase FMS1-spermine complex and demonstrate a major role for residues His82 and Lys367 in substrate binding and catalysis. In addition, the SMO enzyme-substrate complex highlights the presence of an active site pocket with highly polar characteristics, which may explain the different substrate specificity of SMO with respect to APAO and provide the basis for the design of specific inhibitors for SMO and APAO.
Mechanistic and structural analyses of the roles of active site residues in yeast polyamine oxidase Fms1: Characterization of the N195A and D94N enzymes
Adachi, Mariya S.,Taylor, Alexander B.,Hart, P. John,Fitzpatrick, Paul F.
, p. 8690 - 8697 (2012)
Flavoprotein Fms1 from Saccharomyces cerevisiae catalyzes the oxidation of spermine in the biosynthetic pathway for pantothenic acid. The same reaction is catalyzed by the mammalian polyamine and spermine oxidases. The active site of Fms1 contains three amino acid residues positioned to interact with the polyamine substrate, His67, Asn195, and Asp94. These three residues form a hydrogen-bonding triad with Asn195 being the central residue. Previous studies of the effects of mutating His67 are consistent with that residue being important both for interacting with the substrate and for maintaining the hydrogen bonds in the triad [Adachi, M. S., Taylor, A. B., Hart, P. J., and Fitzpatrick, P. F. (2012) Biochemistry 51, 4888-4897]. The N195A and D94N enzymes have now been characterized to evaluate their roles in catalysis. Both mutations primarily affect the reductive half-reaction. With N 1-acetylspermine as the substrate, the rate constant for flavin reduction decreases ～450-fold for both mutations; the effects with spermine as the substrate are smaller, 20-40-fold. The kcat/Kamine- and kcat-pH profiles with N1-acetylspermine are only slightly changed from the profiles for the wild-type enzyme, consistent with the pKa values arising from the amine substrate or product and not from active site residues. The structure of the N195A enzyme was determined at a resolution of 2.0 ?. The structure shows a molecule of tetraethylene glycol in the active site and establishes that the mutation has no effect on the protein structure. Overall, the results are consistent with the role of Asn195 and Asp94 being to properly position the polyamine substrate for oxidation.
Mechanistic studies of human spermine oxidase: Kinetic mechanism and pH effects
Adachi, Mariya S.,Juarez, Paul R.,Fitzpatrick, Paul F.
, p. 386 - 392 (2010)
In mammalian cells, the flavoprotein spermine oxidase (SMO) catalyzes the oxidation of spermine to spermidine and 3-aminopropanal. Mechanistic studies have been conducted with the recombinant human enzyme. The initial velocity pattern in which the ratio between the concentrations of spermine and oxygen is kept constant establishes the steady-state kinetic pattern as ping-pong. Reduction of SMO by spermine in the absence of oxygen is biphasic. The rate constant for the rapid phase varies with the substrate concentration, with a limiting value (k3) of 49 s-1 and an apparent K d value of 48 μMat pH8.3. The rate constant for the slow step is independent of the spermine concentration, with a value of 5.5 s-1, comparable to the kcat value of 6.6 s-1. The kinetics of the oxidative half-reaction depend on the aging time after the spermine and enzyme are mixed in a double-mixing experiment. At an aging time of 6 s, the reaction is monophasic with a second-order rate constant of 4.2 mM-1 s-1. At an aging time of 0.3 s, the reaction is biphasic with two second-order constants equal to 4.0 and 40 mM-1 s-1. Neither is equal to the kcat/KO2 value of 13 mM -1 s-1. These results establish the existence of more than one pathway for the reaction of the reduced flavin intermediate with oxygen. The kcat/KM value for spermine exhibits a bell-shaped pH profile, with an average pKa value of 8.3. This profile is consistent with the active form of spermine having three charged nitrogens. The pH profile for k3 shows a pKa value of 7.4 for a group that must be unprotonated. The pKi-pH profiles for the competitive inhibitors N,N′-dibenzylbutane-1,4-diamine and spermidine show that the fully protonated forms of the inhibitors and the unprotonated form of an amino acid residue with a pKa of ～7.4 in the active site are preferred for binding. 2009 American Chemical Society.
A lysine conserved in the monoamine oxidase family is involved in oxidation of the reduced flavin in mouse polyamine oxidase
Henderson Pozzi, Michelle,Fitzpatrick, Paul F.
, p. 83 - 88 (2010)
Lysine 315 of mouse polyamine amine oxidase corresponds to a lysine residue that is conserved in the flavoprotein amine oxidases of the monoamine oxidase structural family. In several structures, this lysine residue forms a hydrogen bond to a water molecule that is hydrogen-bonded to the flavin N(5). Mutation of Lys315 in polyamine oxidase to methionine was previously shown to have no effect on the kinetics of the reductive half-reaction of the enzyme (M. Henderson Pozzi, V. Gawandi, P.F. Fitzpatrick, Biochemistry 48 (2009) 1508-1516). In contrast, the mutation does affect steps in the oxidative half-reaction. The kcat value is unaffected by the mutation; this kinetic parameter likely reflects product release. At pH 10, the kcat/Km value for oxygen is 25-fold lower in the mutant enzyme. The kcat/KO2 value is pH-dependent for the wild-type enzyme, decreasing below a pKa of 7.0, while this kinetic parameter for the mutant enzyme is pH-independent. This is consistent with the neutral form of Lys315 being required for more rapid flavin oxidation. The solvent isotope effect on the kcat/KO2 value increases from 1.4 in the wild-type enzyme to 1.9 in the mutant protein, and the solvent inventory changes from linear to bowed. The effects of the mutation can be explained by the lysine orienting the bridging water so that it can accept the proton from the flavin N(5) during flavin oxidation. In the mutant enzyme the lysine amine would be replaced by a water chain.
Preparation method of spermidine
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Paragraph 0032-0037, (2021/12/07)
The invention relates to the technical field of preparation of compounds, in particular to a preparation method of spermidine. The method comprises the following steps: S1, reacting 2-pyrrolidone and di-tert-butyl dicarbonate ester in an organic solvent by using 4-dimethylaminopyridine as a catalyst to obtain 1-(tert-butoxycarbonyl)-2-pyrrolidone; s2, dissolving the obtained 1-(tert-butoxycarbonyl)-2-pyrrolidone in a solvent, adding the solution into a 1, 3-propane diamine solution, carrying out stirring and reaction until the reaction is finished, and carrying out post-treatment to obtain [3-(3-amino-alanyl carbamoyl) propyl]-tert-butyl carbamate; s3, dissolving in a solvent, reducing by a reducing agent, and performing post-treatment to obtain [4-(3-amino-propylamino) butyl]-tert-butyl carbamate; and S4, dissolving in a solvent, dropwise adding hydrochloric acid for reaction to remove a protecting group, and after the reaction, further treating to obtain the spermidine. The raw materials such as pyrrolidone, 1, 3-propane diamine and the like are common chemical raw materials, the cost is relatively low, the reaction conditions are relatively mild, and the potential safety hazard in the reaction is relatively low.
Synthesis process of spermidine and intermediate thereof
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Paragraph 0054-0059, (2021/10/27)
The invention provides a synthesis process of spermidine (I) which takes 3 -t-butyloxycarbonyl V (N -) and -1-t-butyloxycarbonyl 4 - IV butanediamine (N -) as a raw material to obtain the spermidine trihydrochloride (-4 - II III) by reductive amination reaction, and provides a novel method for chemical synthesis of spermidine by liberation of the protecting group from the intermediate I (3 -) to obtain the spermidine (III) II.
Method for preparing spermidine
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Paragraph 0057; 0072-0074, (2019/01/14)
The invention belongs to the field of chemical synthesis and particularly relates to a method for preparing spermidine. The method comprises the steps that the spermidine is prepared through the mainprocess steps of reduction, amino protection and the like after reaction of amino-1-propanol and butyrolactone serving as raw materials. The method has the advantages of being simple in operation, high in product quality, high in yield and the like.
Metabolism of N-alkylated spermine analogues by polyamine and spermine oxidases
Haekkinen, Merja R.,Hyvoenen, Mervi T.,Auriola, Seppo,Casero Jr., Robert A.,Vepsaelaeinen, Jouko,Khomutov, Alex R.,Alhonen, Leena,Keinaenen, Tuomo A.
, p. 369 - 381 (2010/11/18)
N-alkylated polyamine analogues have potential as anticancer and antiparasitic drugs. However, their metabolism in the host has remained incompletely defined thus potentially limiting their utility. Here, we have studied the degradation of three different spermine analogues N,N′-bis-(3-ethylaminopropyl)butane-1,4-diamine (DESPM), N-(3-benzyl-aminopropyl)-N′-(3-ethylamino-propyl)butane-1,4-diamine (BnEtSPM) and N,N′-bis-(3-benzylaminopropyl)butane-1,4-diamine (DBSPM) and related mono-alkylated derivatives as substrates of recombinant human polyamine oxidase (APAO) and spermine oxidase (SMO). APAO and SMO metabolized DESPM to EtSPD [Km(APAO) = 10 μM, kcat(APAO) = 1.1 s -1 and Km(SMO) = 28 μM, kcat(SMO) = 0.8 s-1, respectively], metabolized BnEtSPM to EtSPD [Km(APAO) = 0.9 μM, kcat(APAO) = 1.1 s-1 and Km(SMO) = 51 μM, kcat(SMO) = 0.4 s-1, respectively], and metabolized DBSPM to BnSPD [Km(APAO) = 5.4 μM, k cat(APAO) = 2.0 s-1and Km(SMO) = 33 μM, kcat(SMO) = 0.3 s-1, respectively]. Interestingly, mono-alkylated spermine derivatives were metabolized by APAO and SMO to SPD [EtSPM Km(APAO) =16 μM, kcat(APAO) = 1.5 s -1; Km(SMO) = 25 μM, kcat(SMO) = 8.2 s -1; BnSPM Km(APAO) = 6.0 μM, kcat(APAO) = 2.8 s-1; Km(SMO) =19 μM, kcat(SMO) = 0.8 s-1, respectively]. Surprisingly, EtSPD [Km(APAO) = 37 μM, kcat(APAO) = 0.1 s-1; Km(SMO) =48 μM, kcat(SMO) = 0.05 s-1] and BnSPD [Km(APAO) = 2.5 μM, kcat(APAO) = 3.5 s-1; Km(SMO) =60 μM, kcat(SMO) = 0.54 s-1] were metabolized to SPD by both the oxidases. Furthermore, we studied the degradation of DESPM, BnEtSPM or DBSPM in the DU145 prostate carcinoma cell line. The same major metabolites EtSPD and/or BnSPD were detected both in the culture medium and intracellularly after 48 h of culture. Moreover, EtSPM and BnSPM were detected from cell samples. Present data shows that inducible SMO parallel with APAO could playanimportant roleinpolyamine based drug action, i.e. degradation of parent drug and its metabolites, having significant impact on efficiency of these drugs, and hence for the development of novel N-alkylated polyamine analogues. Springer-Verlag 2009.
Protein S-thiolation by glutathionylspermidine (Gsp): The role of Escherichia coli Gsp synthetase/amidase in redox regulation
Chiang, Bing-Yu,Chen, Tzu-Chieh,Pai, Chien-Hua,Chou, Chi-Chi,Chen, Hsuan-He,Ko, Tzu-Ping,Hsu, Wen-Hung,Chang, Chun-Yang,Wu, Whei-Fen,Wang, Andrew H.-J.,Lin, Chun-Hung
experimental part, p. 25345 - 25353 (2011/04/17)
Certain bacteria synthesize glutathionylspermidine (Gsp), from GSH and spermidine. Escherichia coli Gsp synthetase/amidase (GspSA) catalyzes both the synthesis and hydrolysis of Gsp. Prior to the work reported herein, the physiological role(s) of Gsp or how the two opposing GspSA activities are regulated had not been elucidated. We report that Gsp-modified proteins from E. coli contain mixed disulfides of Gsp and protein thiols, representing a new type of post-translational modification formerly undocumented. The level of these proteins is increased by oxidative stress. We attribute the accumulation of such proteins to the selective inactivation of GspSA amidase activity. X-ray crystallography and a chemical modification study indicated that the catalytic cysteine thiol of the GspSA amidase domain is transiently inactivated byH 2O2 oxidation to sulfenic acid, which is stabilized by a very short hydrogen bond with a water molecule. We propose a set of reactions that explains how the levels of Gsp and Gsp S-thiolated proteins are modulated in response to oxidative stress. The hypersensitivities of GspSA and GspSA/glutaredoxin null mutants toH2O2 support the idea that GspSA and glutaredoxin act synergistically to regulate the redox environment of E. coli.
Bisimide compounds
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, (2008/06/13)
A compound of formula (I) STR1 in which: X and Y, which may be identical or different, represent hydrogen or halogen or alkyl, trihaloalkyl, alkoxy, hydroxyl, cyano, nitro, amino, alkylamino, or dialkylamino, Z represent a linear or branched C4 to C12 alkylene chain in which one or more --CH2 -- are optionally replaced by any one of the following atoms or groups: --NR--, --O--, --S--, --SO--, --SO2 --, or --CONH--, A forms, with two adjacent carbon atoms of the phenyl ring, a phenyl, naphthyl or tetrahydronaphthyl ring or a heterocycle, STR2 represents any one of the groups as defined in the description, its optical isomers and its addition salts with a pharmaceutically-acceptable acid or base, and medicinal products containing the same which are useful as anticancer agents.
