The MAO-B catalyzed metabolic bioactivation of the parkinsonian inducing agent 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to generate the neurotoxic 1-methyl-4-phenylpyridinium species (MPP+) is well documented. The N-cyclopropyl analog (CPTP) of MPTP is a mechanism based inactivator of MAO-B which presumably is processed by a single electron transfer (SET) pathway to generate a bioalkylating species. These results have prompted us to study how the cytochromes P450, the major liver drug metabolizing oxidases, interact with N-cyclopropyl analogs of MPTP. HPLC with diode array detection and LC-electrosprary ionization mass spectrometry (LC-ESI/MS) based methods have been developed for metabolite detection and characterization. From the UV spectral data and pseudomolecular ion species observed by LC-ESI/MS, we have identified N-oxide, C-hydroxylated, and pyridinium metabolites. For the trans-1-(2-phenylcyclopropyl) analog, cinnamaldehyde and p-hydroxycinnamaldehyde also were characterized. Incubation of CPTP and its derivatives with cDNA expressed human hepatic cytochrome P450 has shown that CYP2D6 catalyzes the formation of cinnamaldehyde, the N-descyclopropyl, pyridinium and hydroxylated products. CYP3A4 is responsible for the formation of the N-descyclopropyl and pyridinium species and cinnamaldehyde but it does not mediate any hydroxylation reactions. Since both the a-carbon oxidation and N-descyclopropylation transformations are mediated by a single enzyme (either CYP2D6 or CYP3A4), we propose a common intermediate for both pathways, namely the cyclopropylaminyl radical cation generated by the SET pathway. This intermediate partitions between the a-carbon oxidation pathway leading to the dihydropyridinium and pyridinium species and the ring opening pathway leading to the N-descyclopropyl metabolite and aldehyde species. The phenyl substituent on the cyclopropyl ring stabilizes the ring opened distonic radical cation and favors the ring opening pathway and results in the formation of less of the pyridinium species. The proton and methyl substituents on the cyclopropyl ring favor the a-carbon oxidation pathway and increased amounts of the pyridinium species are formed. / Master of Science

京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第19407号 / 工博第4123号 / 新制||工||1636(附属図書館) / 32432 / 京都大学大学院工学研究科合成・生物化学専攻 / (主査)教授 吉田 潤一, 教授 松田 建児, 教授 松原 誠二郎 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DFAM

<p>      Chemical reactions in micron-sized droplets under ambient conditions are often orders of magnitude faster than the equivalent bulk reactions due to the large interfacial effects. The investigation of the underlying mechanisms driving the unique surface chemistry of droplets, as well as their applications and implications in synthesis, has garnered considerable interest. This dissertation delves into three key subtopics: (1) Exploring partial solvation as a mechanism for accelerating reactions in microdroplets, (2) Investigating the spontaneous oxidation and reduction of heteroatom double bonds induced by water radical cations and anions generated from water, and (3) Examining the role of oxazolone intermediates in prebiotic peptide synthesis and the emergence of homochirality in living systems.</p> <p>      Chemical reactions can be accelerated in microdroplets but with previously unclear mechanisms. Here we report a systematic study of organic reactions of common types in microvolumes and compare their rates with those in bulk solution. The observed interfacial area effect, molecularity effect and solvent effect provided experimental evidence for partial solvation at gas/liquid interface as one of the major contributors to the observed more than 10⁴-fold acceleration in microdroplets.</p> <p>      Recent spectroscopic results as well as computations demonstrate the existence of a strong electric field at aqueous droplet surfaces, which can result in microdroplet-specific reactions, especially their intrinsic redox properties. Spontaneous oxidation or reduction without external oxidants or reductants has been reported. One explanation for the existence of active species is dissociation of the radical cation/anion pair (H₂O^+∙/ H₂O^-∙), recently argued to occur in pure bulk water, to provide the free radical cation and radical anion. In this work, we reported spontaneous oxidation of heteroatom double bonds (e.g. sulfone to sulfonic acid, ketone to carboxylic acid) in non-aqueous microdroplets containing traces of water (<1%). Meanwhile, the simultaneous oxidation and reduction of several phosphonates was discovered, supporting the radical pair as the source of reactive species in water microdroplets.</p> <p>      One implication of microdroplet chemistry lies in its connection to prebiotic synthesis. Peptide formation from amino acids is thermodynamically unfavorable but a recent study provided evidence that the reaction occurs at the air/solution interfaces of aqueous microdroplets. Here we show that (i) the suggested amino acid complex in microdroplets undergoes dehydration to form oxazolone; (ii) addition of water to the oxazolone forms the dipeptide; and (iii) reaction of oxazolone with other amino acids forms tripeptides. Furthermore, the chirality of the reacting amino acids is preserved in the oxazolone, and strong chiral selectivity is observed when converting the oxazolone to tripeptide. This last fact ensures that optically impure amino acids will undergo chain extension to generate homochiral peptides. Peptide formation in bulk by wet-dry cycling shares a common pathway with the microdroplet reaction, both involving the oxazolone intermediate.</p>

<p> The organic ions studied in this thesis were generated in the rarefied gas phase of the mass spectrometer by electron ionization of selected precursor molecules. The characterization of their structure and reactivity was probed by using a variety of tandem mass spectrometry techniques. These include metastable ion spectra to probe the dissociation chemistry of the low energy ions and collision experiments to establish the atom connectivity of the ions. The technique of neutralization-reionization mass spectrometry (NRMS) was used to probe the structure and stability of the neutral counterparts of the ions. Computational results involving the CBS-QB3 model chemistry formed an integral component in the interpretation of the experimental findings.</p> <p> The above approach was used to study proton-transport catalysis in the formaldehyde elimination from low energy 1,3-dihydroxyacetone radical cations. Solitary ketene-water ions, CH2=C(=O)OH2·+, do not readily isomerize into its more stable isomer, CH2=C(OH)2·+. A mechanistic analysis using the CBS-QB3 model chemistry shows that metastable 1,3-dihydroxyacetone radical cations will rearrange into hydrogen-bridged radical cations [CH2C(=O)O(H)-H•••OCH2]·+, where the CH2=O will catalyze the transformation of CH2=C(=O)OH2·+ into CH2=C(OH)2·+.</p> <p> Metastable pyruvic acid radical cations, CH3C(=O)COOH·+, have been shown to undergo decarboxylation to yield m/z 44 ions, C2H4O·+, in competition with the formation of CH3C=O+ + COOH· by direct bond cleavage. Collision induced dissociation experiments agree with an earlier report that oxycarbene ions CH3COH·+ are formed but they also suggest the more stable isomer CH3C(H)=O·+ may be co-generated. Using the CBS-QB3 model chemistry, a mechanism is proposed to rationalize these results.</p> <p> Next, the isomeric ions CH3O-P=S·+ and CH3S-P=O·+ were characterized and differentiated by tandem mass spectrometry. Metastable CH3O-P=S·+ and CH3S-P=O·+ ions both spontaneously lose water to yield m/lz 74 cyclic product ion [-S-CH=]P·+. Using the CBS-QB3 model chemistry a mechanism is proposed for the water loss from CH3O-P=S·+ and CH3S-P=O·+. Our calculations also show that these two isomers communicate via a common intermediate, the distonic ion CH2S-P-OH·+, prior to the loss of water.</p> <p> The final component of this work details the computational study addressing the long standing question on the mechanism for the water elimination from metastable ethyl acetate radical cations. The CBS-QB3 results show that low energy ethyl acetate ions isomerize into ionized 4-hydroxy-2-butanone prior to the loss of water.</p> / Thesis / Master of Science (MSc)

Macrocyclic ligand-complexed transition metal-oxo intermediates are the active oxidizing species in a variety of important biological and catalytic oxidation reactions. Many transition metal catalysts have been designed to mimic the predominant oxidation catalysts in nature, namely the cytochrome P450 enzymes. Iron porphyrin complexes have been the center of research as catalysts. In this study 5,10,15,20- tetramesitylporphyrin (H2TMP) and its corresponding iron complexes FeIII(X)TMP (X= Cl, ClO4, ClO3, NO3, NO2, and BrO3) have been successfully synthesized and fully characterized by UV-vis and NMR spectroscopies. For the catalytic selective oxidation of organic sulfides, the potential of iron(III) porphyrin complexes with iodobenzene diacetate [PhI(OAc)2] have been investigated. Iodobenzene diacetate was found to be an efficient oxygen source in the iron(III) porphyrin-catalyzed oxidation of sulfides to sulfoxides. Iron(III) porphyrin catalysts show an excellent conversion and selectivity for the sulfoxidation reactions. Reaction conditions and environments that effect the catalytic sulfoxidation including solvent, catalytic amount, axial ligand, water, and thioanisole substrates, have been investigated to identify the optimal conditions and the substrate scope. Under optimized conditions, excellent substrate conversions (up to 100%) as well as product selectivies (sulfoxide:sulfone > 95:5) have been achieved. To probe the nature of the oxidizing species in above catalytic sulfoxidations, iron(IV)-oxo porphyrin radical cations model of Compound I were chemically produced from the corresponding iron(III) tetramesitylporphyrin precursors with excess amounts of PhI(OAc)2 (20-50 equivalents) in CH3CN solvent. All O=FeIV(X)TMP·+ (X= Cl, ClO4, ClO3, and NO3) show weaker Soret band and broader Q band that are characteristic of Compound I analogues. A new photochemical method that led to generation of the iron(IV)-oxo porphyrin radical cations was also successfully developed. Iron(IV)-oxo porphyrin radical cations were generated by irradiation of iron(III) porphyrin chlorate or bromate complexes that result in heterolytic cleavage of the O-X bond in the axial ligand.

Polycyclic aromatic azomethine ylide (PAMY) is a versatile building block for the bottom-up construction of unprecedented nitrogen-containing polycyclic aromatic hydrocarbons (N-PAHs). Here, we demonstrate the 1,3-dipolar cycloaddition between PAMY and 1,4-diphenylbut-2-yne-1,4-dione as well as the subsequent condensation reaction with hydrazine, which led to synthesis of unique N-PAHs with a phenyl-substituted pyrrolopyridazine core (PP-1 and PP-2). The molecular structures of pristine PP-1 and tert-butyl-substituted PP-2 were verified by NMR and mass spectroscopy. Moreover, the structure of PP-2 was unambiguously elucidated by X-ray single crystal analysis. The optoelectronic properties were investigated by solvent-dependent UV-Vis absorption and fluorescence emission spectroscopy as well as cyclic voltammetry. Additionally, the density functional theory (DFT) calculations exhibited a push-pull behavior for PP-1 and PP-2. Furthermore, the in situ EPR/UV-Vis-NIR spectroelectrochemistry allowed the detailed insight into the spectroscopic properties and spin distribution of radical cation species of PP-2.

<p>High resolution tandem mass spectrometry (MSⁿ) coupled with various separation techniques, such as high-performance liquid chromatography (HPLC) and gas chromatography (GC), is widely used to analyze mixtures of unknown organic compounds. In a mass spectrometric analysis, analytes of interest are at first transferred into the gas phase, ionized (protonated or deprotonated) and introduced into the instrument. Tandem mass spectrometric experiments may then be used to gain insights into structure and reactivity of the analyte ions in the gas phase. The tandem mass spectral data are often compared to those reported in external databases. However, the tandem mass spectra obtained for protonated analytes may be markedly different from those in external databases because protonation site manifested during a mass spectrometric experiment can be affected by the ionization technique, ionization solvents and condition of the ion source. This thesis focuses on investigating the effects of instrumental conditions and analyte concentrations on the protonation sites of 4-aminobenzoic acid. Reactivities of radical species were also investigated. A modified bracketing method was developed and proton affinities of a series of mono- and biradicals of pyridine were measured. In another study, a <i>para</i>-benzyne analog was generated in both solution and the gas phase and its reactivities towards various neutral reagents in the gas phase were compared to those in solution.</p> <p> Chapter 2 discusses the fundamental aspects of the instruments used in this research. In chapter 3, the effects of residual moisture in linear quadrupole ion trap on the protonation sites of 4-aminobenzoic acid are considered. Chapter 4 focuses on the use of gas-phase ion-molecule reactions with trimethoxymethylsilane (TMMS) for the identification of the protonation sites of 4-aminobenzoic acid. Further, the effects of analyte concentration on the protonation sites of 4-aminobenzoic acid are considered. Chapter 5 introduces a modified bracketing method for the experimental determination of proton affinities of a series of pyridine-based mono- and biradicals. In chapter 6, successful generation of <i>para</i>-benzynes in solution is discussed. The reactivity of a <i>para</i>-benzyne analog, 1,4-didehydrophenazine, is compared to its reactivity in the gas phase.</p>

No description available.