NL2021113B1 - Synthesis of aromatic epoxide derived compounds - Google Patents

Synthesis of aromatic epoxide derived compounds Download PDF

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NL2021113B1
NL2021113B1 NL2021113A NL2021113A NL2021113B1 NL 2021113 B1 NL2021113 B1 NL 2021113B1 NL 2021113 A NL2021113 A NL 2021113A NL 2021113 A NL2021113 A NL 2021113A NL 2021113 B1 NL2021113 B1 NL 2021113B1
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aromatic
production process
compound
epoxide
oxidoreductase
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NL2021113A
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Hollmann Frank
Zhang Wuyuan
Hamdy Hamed Younes Sabry
Tieves Florian
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Univ Delft Tech
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Priority to PCT/NL2019/050361 priority patent/WO2019240579A1/en
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    • C07C215/44—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having amino groups or hydroxy groups bound to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton bound to carbon atoms of the same ring or condensed ring system
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    • C07C2602/04—One of the condensed rings being a six-membered aromatic ring
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    • C07C2603/40—Ortho- or ortho- and peri-condensed systems containing four condensed rings
    • C07C2603/42—Ortho- or ortho- and peri-condensed systems containing four condensed rings containing only six-membered rings
    • C07C2603/50—Pyrenes; Hydrogenated pyrenes

Abstract

The invention provides a production process for the production of an aromatic epoxide derived compound, the production process comprising: (i) an epoxidation stage comprising: reacting an aromatic compound and a peroxide in the presence of an oxidoreductase, resulting in an aromatic epoxide, Wherein the aromatic compound comprises an aromatic ring structure, wherein the oxidoreductase catalyzes epoxidation of the aromatic ring structure, and wherein the oxidoreductase is selected from the group consisting of oxygenases, peroxygenases, oxidases, and peroxidases; (ii) a ring-opening stage comprising: reacting the aromatic epoxide and a nucleophile, resulting in the aromatic epoxide derived compound.

Description

FIELD OF THE INVENTION
The invention relates to a production process for an aromatic epoxide derived compound. The invention further relates to products formed in such production process.
BACKGROUND OF THE INVENTION
Production processes for aromatic epoxides (‘‘arene oxides”) are known in the art. For example, JPS6347711B2 describes conducting a reaction between a polycyclic aromatic compound such as phenanthrene, pyrene or benzo[a]pyrene and a hypochlorite salt such as sodium hypochlorite under phasetransfer conditions in an inert solvent such as chlorinated or nitrated hydrocarbon such as methylene dichloride or chloroform, at 35-55 °C under reflux to give an arene oxide. The phase transfer condition is made by using tetra-n-butyl ammonium chloride as a catalyst.
SUMMARY OF THE INVENTION
Aromatic epoxides (“arene oxides”) may be valuable building blocks for chemical synthesis. Next to a NIH rearrangement (also “NIH shift” named for the National Institutes of Health where the rearrangement was first reported) into corresponding aromatic alcohols, also nucleophilic epoxide ringopening may lead to a broad range of valuable aromatic epoxide derived compounds.
Their synthetic value, however, may be rather limited due to tedious, multistep synthetic protocols. Using (classical) chemical methods, direct epoxidation of aromatics may only be possible under extreme and/or harsh conditions, such as substantially acidic conditions, and therefore aromatic epoxides may not be accessible for preparative synthesis. Furthermore, the (classical) chemical methods may result in a large amount of waste, such as a large amount of halogen waste.
P450 monooxygenases, however, may directly epoxidise aromatic xenobiotics. Also with the mechanistically related peroxygenases, aromatic epoxide intermediates may be produced. The intermediate aromatic epoxide, however, may be considered to be too unstable to be of any synthetic use except as precursor for the corresponding aromatic alcohols (via NIH rearrangement).
Aromatic epoxides may be potentially useful chemical building blocks. However, known (chemical) synthesis methods, that may be used for the production of aromatic epoxides, may be considered tedious and dangerous.
To the best of our knowledge, there is a very limited exploration and use of aromatic epoxides as substrates in chemical syntheses, if any.
Aromatic epoxides may, however, also be formed in biological systems. For example, the gene APO1 of Agrocybe aegerita codes for an unspecific peroxygenase (also: “aromatic peroxygenase”). The unspecific peroxygenase may be able to catalyze a reaction between an aromatic compound and a peroxide leading to the formation of an aromatic alcohol. The aforementioned reaction may occur via an aromatic epoxide intermediate undergoing a NIH rearrangement. Such aromatic epoxide may generally be regarded to spontaneously undergo the NIH rearrangement leading to the formation of the aromatic alcohol. Indeed, leading biochemical databases such as KEGG, BRENDA, and Biocyc relate the activity of an unspecific peroxygenase (EC: 1.11.2.1) to the introduction of a hydroxyl-group rather than to the formation of an epoxide.
Hence, the application of aromatic epoxides as building blocks for further chemical syntheses may suffer from difficulties in their synthesis as well as an (alleged) lack of stability leading to the NIH rearrangement.
Hence, it is an aspect of the invention to provide an alternative production process for an aromatic epoxide derived compound (also “aromatic epoxide derived chemical”), which preferably further at least partly obviates one or more of above-described drawbacks. The present invention may have as object to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.
We have surprisingly discovered, however, that aromatic epoxides may be sufficiently stable for a nucleophilic attack using, for example, azide, thereby opening up the potential for the synthesis of a wide range of aromatic epoxide derived compounds.
The present invention relates to the formation of aromatic epoxide derived compounds, via the formation of an aromatic epoxide using an oxidoreductase, and a subsequent reaction involving a nucleophile - such as an azide ion - instead of the spontaneous isomerization to an aromatic alcohol.
Therefore, in a first aspect the invention provides a production process for the production of an aromatic epoxide derived compound, the production process comprising: (i) an epoxidation stage comprising: reacting an aromatic compound and a peroxide in the presence of an oxidoreductase, resulting in an aromatic epoxide, wherein the aromatic compound comprises an aromatic ring structure, wherein the oxidoreductase catalyzes epoxidation of the aromatic ring structure, and wherein the oxidoreductase is especially selected from the group consisting of oxygenases, peroxygenases, oxidases, and peroxidases; (ii) a ring-opening stage comprising: reacting the aromatic epoxide and a nucleophile, resulting in the aromatic epoxide derived compound.
The invention as described herein may provide a wealth of novel synthesis opportunities as aromatic epoxides may have received little attention so far due to an alleged lack of stability. The ring-opening-stage (also: nucleophilic attack stage) following the epoxidation of the ring structure may be carried out by various nucleophiles, resulting in a variation of possible aromatic epoxide derived compounds for each potential starting aromatic compound. The aromatic epoxide derived compound may be further reacted with one or more reactants, resulting in a second aromatic epoxide derived compound. Several non-limiting examples hereof are highlighted in the embodiments.
The term “aromatic epoxide derived compound” is hereinafter especially used to refer both to the product of the ring-opening-stage, as well as to a “second aromatic epoxide derived compound”, i.e., any product of the production process as described herein may be referred to as “aromatic epoxide derived compound”. The term “second aromatic epoxide derived compound” is specifically used herein to refer to a compound resulting from one or more additional reactions following the ring-opening-stage.
The enzymatic epoxidation step may result in a substantially enantiomerically pure aromatic epoxide, which may further result in a substantially enantiomerically pure aromatic epoxide derived compound. The term “enantiomer” refers to a member of a pair of non-superimposable mirrorimaged compounds. As two enantiomers often have different biological functions, the relative abundance of one enantiomer is typically reported as enantiomeric excess (ee), wherein ee = | F+ - F.|, i.e., the absolute difference between the mole fractions of a first enantiomer (F+) and the corresponding second enantiomer (F_). A substantially enantiomerically pure compound may have an enantiomeric excess (ee) selected from the range of 0.8-1, such as from the range of 0.9-1, especially from the range of 0.95-1. The term “enantiomerically pure” refers to a compound (sample) consisting of a single enantiomer, i.e., a compound (sample) having an ee equal to 1. In embodiments, the production process as described herein may provide enantiomerically pure aromatic epoxide derived compounds.
The epoxidation stage comprises the oxidoreductase catalyzed epoxidation of an aromatic compound by a peroxide, resulting in the formation of an aromatic epoxide. In embodiments, the epoxidation stage may comprise reacting the aromatic compound and the peroxide in the presence of a first solvent for the aromatic compound. The first solvent may comprise one or more of a nonpolar solvent, a polar aprotic solvent, a polar protic solvent, and/or a supercritical solvent. In further embodiments, the first solvent may comprise one or more of a nonpolar solvent a polar aprotic solvent, or a supercritical solvent. In alternative embodiments, the first solvent may comprise an aqueous solvent.
In embodiments wherein the first solvent comprises a nonpolar solvent, the first solvent may comprise one or more solvents selected from the group consisting of pentane, cyclopentane, hexane, cyclohexane, benzene, toluene, 1,4-dioxane, chloroform, diethyl ether, and dichloromethane.
In embodiments wherein the first solvent comprises a polar aprotic solvent, the first solvent may comprise one or more solvents selected from the group consisting of tetrahydrofuran, ethyl acetate, acetone, dimethylformamide, acetonitrile, dimethyl sulfoxide, nitromethane, and propylene carbonate.
In embodiments wherein the first solvent comprises a polar protic solvent, the first solvent may comprise one or more solvents selected from the group consisting of formic acid, n-butanol, isopropyl alcohol, n-propanol, ethanol, methanol, acetic acid, and water.
In embodiments wherein the first solvent comprises a supercritical solvent, the first solvent may comprise e.g., scCO2 (super critical CO2).
In embodiments, the first solvent may comprise the aromatic compound. In further embodiments, the first solvent may consist of the aromatic compound. In alternative embodiments, the first solvent may be chosen such that the oxidoreductase does not catalyze epoxidation of the first solvent.
The epoxidation stage may comprise reacting the aromatic compound and the peroxide in the presence of an epoxidation reaction mixture, wherein the epoxidation reaction mixture comprises one or more of the aromatic compound, the peroxide, the peroxygenase, the first solvent, a first buffer, and a first salt. The epoxidation reaction mixture may be configured to provide specific reaction conditions, such as a specific pH and/or ionic strength. Hence, in embodiments the epoxidation stage may comprise reacting the aromatic compound and the peroxide at a pH selected from the range of 5.0-8.5.
In embodiments, the epoxidation stage may comprise reacting the aromatic compound and the peroxide at about a first temperature. In further embodiments, the first temperature may especially be selected from the range of 0-45 °C, more especially from the range of 20-45 °C. Hence, in embodiments the epoxidation stage may comprise reacting the aromatic compound and the peroxide at a temperature selected from the range of 0-45 °C.
An epoxide is especially defined as a cyclic ether with a threeatom ring structure, e.g., a compound with the formula R1-O-R2, or including a group with such formula, wherein Ri and R2 are also covalently bonded. Hence, an epoxide comprises an epoxide group of an O and 2 other atoms, wherein each of the three atoms is covalently bonded to the other two. The term “aromatic epoxide” herein especially refers to a compound obtainable by the introduction of an epoxide group in an aromatic ring structure, i.e., both atoms covalently bonded to oxygen after epoxidation were part of an aromatic ring structure prior to epoxidation. Hence, the oxidoreductase may catalyze the epoxidation of the aromatic ring structure. Ri and R2 are independently selected from carbon comprising (organic) groups, wherein a carbon atom is bonded to the O atoms of the respective organic compounds of the epoxide. Hence, an epoxide may also be defined as a compound with the formula (Ra,Rb)Cl-O-C2(Rc,Rd), or including a group with such formula, wherein C1 and C2 are covalently bonded, and wherein Ra, Rb, Rc, and Rd are each independently e.g. selected from H and a hydrocarbon.
Aromatic compounds may be well known, and may be very stable. In general, a compound has to meet four criteria in order to be considered aromatic: (i) the compound comprises a delocalized conjugated π system (commonly depicted as alternating single and double bonds), (ii) the compound comprises a coplanar structure, with all atoms contributing to the conjugated π system in the same plane, (iii) the compound comprises an aromatic ring structure comprising the atoms contributing to the delocalized conjugated π system in one or more rings, (iv) the compound comprises 4n+2 delocalized π electrons, wherein n is a non-negative integer (Hiickel's rule).
The aromatic compound herein especially comprises an aromatic ring structure. In embodiments, the aromatic compound may comprise one or more of C, N, P, O or any other (ring) element. In further embodiments, the aromatic compound may comprise an aromatic hydrocarbon. In general, in embodiments the aromatic compound comprises at least H and C. However, embodiments wherein the aromatic compound does not comprise H and/or C are not excluded. For example, the aromatic compound may comprise pentazole (HN5), or hexachlorobenzene. In embodiments, the aromatic ring structure may comprise one or more different elements, especially one or more of C, N, O, S, Sn, Si, B, Se and P. In specific embodiments, the aromatic ring structure may comprise two or more different elements, i.e., the aromatic ring structure may comprise a heterocyclic ring structure. In alternative embodiments, the aromatic ring structure may consist of a plurality of atoms of the same type of one element, especially of C or N, i.e., in embodiments the aromatic ring structure may comprise a homocyclic ring structure. Hence, in embodiments, the aromatic ring structure may consist of carbon atoms.
The term “aromatic ring structure’’ used herein especially refers to the ring structure and not to the side groups. Hence, the aromatic compound comprises an aromatic ring stracture having side groups. For example, the aromatic ring structures of benzene and toluene both consists of six C, whereas the aromatic ring structure of naphthalene consists of ten C, and the aromatic ring structure of furan consists of four C and one O.
In embodiments, the side groups of the aromatic ring structure may comprise one or more different elements. Especially, the aromatic ring structure may have one or more side groups independently selected from the group comprising H, CH3, Cl, Br, I, OCH3, CF3, F, CN, NO2, NH2, CONH2, COCH3, NHR, OCH2CH3, OH. Especially, one or more side groups of the aromatic ring stracture may consist of H, such as two or more (adjacent) side groups. More especially, all side groups of the aromatic ring stracture may consist of H.
In specific embodiments, the aromatic compound may comprise one or more of naphthalene, 1-chloronaphthalene, benzene, aniline, toluene, phenanthrene, pyrene, p-nitrophenol, pyridine, dibenzofuran, anthracene, chrysene, fluoranthrene, fluorine, corannulene, coronene, hexahelicene.
The peroxide comprises a compound selected from the group of compounds having the formula Ri-0-0-R2, wherein Ri and R2 refer to side groups comprising any elements, especially side groups comprising C and/or H, more especially side groups consisting of H. Hence, in embodiments the peroxide may comprise H2O2. In further embodiments, the epoxidation step may comprise reacting the aromatic compound and the peroxide in the presence of an H2O2 source, such as one or more compounds selected from the group consisting of carbamide peroxide, sodium percarbonate, ammonium peroxy di sulfate, organic hydroperoxides (e.g. acetone peroxide, acetyl acetone peroxide, acetozone (acetyl benzoyl peroxide), ascaridole, alkenyl peroxides, tert-butyl hydroperoxide, bis(trimethylsilyl) peroxide, cumene hydroperoxide, di-(lnaphthoyl)peroxide, diacetyl peroxide, di-tert-butyl peroxide (dtbp), dimethyldioxirane (dmdo), dioxirane, dipropyl peroxydicarbonate, ethyl hydroperoxide, iodoxy compounds, magnesium monoperoxy-phthalate, methyl ethyl ketone peroxide, methyl isobutyl ketone peroxide, metachloroperoxybenzoic acid (mcpba), tert-butyl peroxy benzoate (tbpb), paramenthane hydroperoxide (pmhp), peracetic acid, performic acid, peroxyacyl nitrates, and peroxybenzoic acid). Hence, in embodiments, the first reaction mixture may further comprise an H2O2 source.
Oxidoreductases are a large class of enzymes generally catalyzing reactions involving the transfer of electrons. Of specific interest herein are those oxidoreductases that catalyze reactions further involving the introduction of O into an organic molecule, such as reactions catalyzed by subclasses of oxidoreductases such as oxidases, peroxidases, oxygenases, and peroxygenases. Hence, in embodiments of the invention the oxidoreductase may be selected from the group consisting of oxidases, peroxidases, oxygenases and peroxygenases. It will be clear to one skilled in the art that the invention is not limited to the use of oxidoreductase (classified as) belonging to any one of the aforementioned subclasses. It will further be clear to one skilled in the art that not each enzyme belonging to the aforementioned subclasses will catalyze the epoxidation of an aromatic ring structure. Hence, it will be clear to one skilled in the art that the invention relates to any oxidoreductase that catalyzes the epoxidation of an aromatic ring structure.
Oxidases may catalyze oxidation-reduction reactions, especially oxidation-reduction reactions involving O2. At least part of the oxidases may catalyze a reaction involving the introduction of an oxygen into an organic compound. For example, cytochrome P450 oxidase may catalyze a monooxygenase reaction of the form RH + 0? + NADPH + H -½ ROH + H.O + NADP+. Similarly, a xanthine oxidase may catalyze a reaction of the form RH +
202 + H2O ROH + 2Ο2’ + 2H+. In specific instances, the introduction of oxygen in an organic compound may occur via an epoxide intermediate.
Peroxidases may typically catalyze reactions of the form R1OOR2 + 2e + 2H’ RiOH + R2OH. At least part of the peroxidases may catalyze a reaction involving the introduction of an oxygen into an organic compound. For example, a peroxidase may catalyze a reaction selected from the group comprising Baeyer-Villiger oxidations, oxidations of styrene derivatives to corresponding ketones, and oxidations of sulfides to sulfoxides and sulfones.
Oxygenases may catalyze reactions involving the transfer of an oxygen atom from O2 to a substrate, especially to an organic compound. Oxygenases may be further classified into (i) monooxygenases, such as the aforementioned cytochrome P450 oxidase, which introduce one oxygen atom into a substrate, and (ii) dioxygenases which introduce both oxygen atoms from O2 into a substrate. In specific instances, the introduction of oxygen may occur via an epoxide intermediate.
Peroxygenases may catalyze reactions involving the transfer of an oxygen atom from a peroxide to a substrate, especially to an organic compound. Hence, peroxygenases may typically catalyze a reaction of the form RiH + R200H RiOH + R2OH, especially wherein R2 consists of H. In specific instances, the introduction of oxygen may occur via an epoxide intermediate.
In embodiments, the oxidoreductase may comprise one or more of a cytochrome P450 enzyme, a heme-dependent and/or a vanadium-dependent oxygenase, a catalase, an unspecific peroxygenase (Enzyme Classification (EC) 1.11.2.1), a peroxidase (EC 1.11.1.7), a chloride peroxidase (EC 1.11.1.10) and a bromide peroxidase (EC 1.11.1.18), especially one or more of an unspecific peroxygenase, a peroxidase, a chloride peroxidase, and a bromide peroxidase, more especially an unspecific peroxygenase.
In embodiments wherein the oxidoreductase comprises an unspecific peroxygenase, the unspecific peroxygenase and/or a gene encoding the unspecific peroxygenase may be derived from one or more organisms selected from the group comprising Agrocybe aegerita, Agrocybe acericola, Agrocybe amara, Agrocybe aivalis, Agrocybe cylindracea, Agrocybe dura,
Agrocybe erebia, Agrocybe farinacea, Agrocybe jinua, Agrocybe molesta, Agrocybe paludosa, Agrocybe parasitica, Agrocybe ped iade s, Agrocybe praecox, Agrocybe putaminum, Agrocybe retiger a, Agrocybe semiorbiculcuis, Agrocybe sororia, Agrocybe vervacti, Coprinellus radians, Coprinellus amphithallus, Coprinellus angulatus, Coprinellus aureogranulatus, CoprineUus bipellis, Coprinellus bisporiger, Coprinellus bisporus, Coprinellus callinus, Coprinellus coiigi-egates, Coprinellus curtus, Coprinellus deliquescens, Coprinellus deminutus, Coprinellus dilectus, Coprinellus disseminatus, Coprinellus domesticus, Coprinellus ellisii, Coprinellus ephemerus, Coprinellus flocculosus, Coprinellus heptemerus, Coprinellus heterosetulosus, Copinellus hiascens, Coprinellus inipatiens, Coprinellus marculentus, Coprinellus mitrinodulisporus, Coprinellus pellucidus, Coprinellus plagioporus, Copriiiellus pyrrhanthes, Coprinellus radians, Coprinellus sassii, Coprinellus sclerocystidiosus, Coprinellus subdis semina tus, Coprinellus subimpatiens, Coprinellus subpurpureus, Coprinellus truncoruni, Coprinellus velatopruinatus, Coprinellus verrucispei-imis, Coprinellus xcnithothrix, Coprinopsis cinerea, Mai'asmius rotula, and Sulfolobus tokodaii. Especially, the unspecific peroxygenases and/or gene encoding the enzyme may be derived from one or more organisms selected from the group consisting οΐ Agrocybe aegerita, Coprinellus radians, Marasmius rotula and Sulfolobus tokodaii.
In embodiments wherein the oxidoreductase comprises a peroxidase, the peroxidase and/or a gene encoding the peroxidase may be derived from one or more organisms selected from the group comprising Acorus calamus, Aedes aegypti, Aggegatibacter actinomycetemcomitaiis, Allium sativum, Arahidopsis thalicnia, Arachis hypogaea, Armoracia rusticana, Arthromyces ramosus, Arundo donax, Beta vulgaris, Bjerkandera adusta, Bos taurus, Brassica napus, Brassica oleracea, Brassica rapa, Bubalus bubali, Butia capitata, Camellia sinensis, Capra hircus, Capsiam anmmm, Catharantus roseus, Chromolaena odorata, Cicer anetinmn, Coprinopsis cinerea, Cucimiis melo, Ciicumis melo var, inodorus, Cynara cardunculiis, Elaeis giiineensis, Elizabethkingia meningoseptica, Escherichia coli, Euphorbia characias, Fagopyrtmi esculentum, Fragaria vesca, Fragaria x ananassa, Glycine max,
Gossypiwv hirsutum, Helianthus annuus, Homo sapiens, Hordeum vulgare, Ipomoea batatas, Ipomoea caniea, Jubaea chilensis, Landoltia punctata, Leptogium saturninum, Malus x domestica, Mentha arvensis, Momordica charantia, Mus musculus, Mycobacterium tuberculosis, Neurospora crassa, Nicotiana sylvestris, Nicotiana tabacum, Oryza sativa, Ovis aries, Pelargonium gim>eolens, Plasmodium falciparum, Pleurotus eryngii, Pleurotus ostreatus, Prunus persico, Pyrococcus furiosus, Raphanus sativus, Ratius non'egicus, Roystonea regia, Ruegeria pomeroyi DSS-3, Sabal minor, Sclerocarya birrea, Scutellaria baicalensis, Senecio scpialidus, Seshania rosfata, Solwmm lycopersicum, Solarium melongena, Sorghum bicolor, Sphagimm magellanicum, Sti-eptomyces thermoviolaceus, Sulfolobus acidocaldarius, Sus scrofa, Trachycarpus fortunei, Triticum aestivum, Vitis vinifera, Washingtonia filifera, and Yersinia pseudotuberculosis.
In embodiments wherein the oxidoreductase comprises a chloride peroxidase, the chloride peroxidase and/or a gene encoding the chloride peroxidase may be derived from one or more organisms selected from the group comprising Caldariomyces fumago, Aspergillus niger, Bazzania tiilobata, Musa paradisiaca, and Streptomyces toyocaensis.
In embodiments wherein the oxidoreductase comprises a bromide peroxidase, the bromide peroxidase and/or a gene encoding the bromide peroxidase may be derived from one or more organisms selected from the group comprising Agocybe aegerita, Ascophyllum nodosum, Corallina officinalis, Corallina pilulifera, Delisea pulchra, Ecldonia stolonifera, Fucus distichus, Gracilaria changii, Homo sapiens, Kappaphycus alvaiezii, Laminaria hyperborea, Macrocystis pyrifera, Ochtodes secundiramea, Pseudomonas fluorescens, Pseudomonas putida, Saccharina latissima, Sti-eptomyces aureofaciens, Sti-eptomyces gi-iseus, Sti-eptomyces venezuelae, and Syiiechococcus sp.
It will be clear to one skilled in the art that the invention is not limited to the use of a native oxidoreductase of any one of the organisms specifically mentioned herein. Rather, the oxidoreductase may comprise an oxidoreductase of an organism not specifically mentioned herein, especially wherein the enzyme is homologous to an oxidoreductase of any one of the mentioned organisms. Alternatively or additionally, the oxidoreductase may comprise a mutant oxidoreductase, such as a mutant oxidoreductase comprising one or more amino acid substitutions, deletions and/or additions relative to a native oxidoreductase, especially wherein the mutant oxidoreductase is specifically designed through protein engineering.
In specific embodiments, the oxidoreductase may comprise a mutant unspecific peroxygenase. For example, the oxidoreductase may comprise one or more of the PaDa-I, the JaWa, and the Solo mutants of the unspecific peroxygenase of Agrocybe aegerita as described in Molina-Espeja et al. 2016 ChemBioChem and in WO2017081355A1.
In embodiments, the oxidoreductase may be provided via one or more micro-organisms producing the oxidoreductase, or via an addition of isolated oxidoreductase. Herein the term “isolated oxidoreductase” refers to biologically, especially microbially, produced oxidoreductase that has been isolated from the production organism. The isolated oxidoreductase may essentially comprise purified oxidoreductase. In general, embodiments of the invention involve the use of isolated oxidoreductase.
In embodiments, the oxidoreductase may be produced by an organism naturally producing the oxidoreductase. Alternatively or additionally, the oxidoreductase may be produced by a genetically modified organism. For example, in an embodiment the unspecific peroxygenase apol gene of Agrocybe aegerita is heterologously expressed in Pichia pastor is X-33, which exports the Apol protein (the unspecific peroxygenase) into a medium, for example into a liquid (growth) medium. In alternative embodiments, the oxidoreductase may be produced by an organism naturally producing the oxidoreductase, especially wherein the organism exports the oxidoreductase into a medium. In further embodiments, the oxidoreductase may be isolated from the medium to obtain an isolated oxidoreductase. For example, the media comprising (microbial) cells and oxidoreductase may be centrifuged such that the cells precipitate while the oxidoreductase remains in the supernatant. The supernatant comprising the oxidoreductase may be used as a crude enzyme preparation in the epoxidation stage, i.e., in embodiments the oxidoreductase may be provided as crude enzyme preparation. The oxidoreductase may also be further purified from the crude enzyme preparation, i.e., in embodiments the oxidoreductase may be provided in purified form. Methods for heterologous gene expression, protein production, protein isolation, and protein purification will be known by a person skilled in the art.
The ring-opening stage comprises reacting the aromatic epoxide (of the epoxidation stage) and a nucleophile, resulting in an aromatic epoxide derived compound. Especially, the ring-opening stage may comprise a nucleophilic substitution reaction at the epoxide group (“ring-opening reaction”), especially at an element bonded to O. The nucleophilic substitution reaction may comprise an SnI or an Sn2 type reaction.
The ring-opening stage may comprise reacting the aromatic epoxide and the nucleophile in the presence of a ring-opening reaction mixture, wherein the ring-opening reaction mixture may comprise one or more of the aromatic epoxide, the nucleophile, a second solvent, a second buffer, a second salt, and a second enzyme. The ring-opening reaction mixture may be configured to provide specific reaction conditions, such as a specific pH and/or ionic strength. In embodiments, the ring-opening reaction mixture may further be configured to catalyze the nucleophilic substitution reaction, especially the second enzyme may catalyze the nucleophilic substitution reaction.
In embodiments, the ring-opening stage may comprise reacting the aromatic epoxide and the nucleophile at about a second temperature. In further embodiments, the second temperature may especially be selected from the range of 0-60°C, more especially from the range of 20-50 °C.
A nucleophile is a chemical compound that can donate an electron pair to an electrophile to form a chemical bond, especially a covalent bond. In embodiments of the invention, the aromatic epoxide, especially the epoxide group, comprises the electrophile. In embodiments, the nucleophile may comprise one or more of N<, CN', SCN’, OCN’, S2‘, ROH, RO’, CT, Br', Γ, hco2; no2; ch3co2; ch3cos; (ch3ch2)3n, (ch3ch2)3p, nh3, h2s, rnh2, RNHR’, R-SH, PhSH, PhSeH, or PhOH, wherein Ph refers to a phenyl group, and wherein R and R’ independently refer to a chemical group comprising any elements but not being directly involved in the reaction.
In embodiments, the production process may further comprise a synthesis stage. The synthesis stage may comprise reacting the aromatic epoxide derived compound with one or more reactants, resulting in a second aromatic epoxide derived compound. Hence, the synthesis stage may comprise a synthesis reaction comprising reacting the aromatic epoxide derived compound with one or more reactants, resulting in a second aromatic epoxide derived compound. Especially, the synthesis stage may comprise a plurality of successive (synthesis) reactions, i.e., the synthesis stage may comprise successively reacting the aromatic epoxide derived compound with one or more reactants.
The synthesis stage may comprise reacting the aromatic epoxide derived compound and the one or more reactants in the presence of a synthesis reaction mixture, wherein the synthesis reaction mixture may comprise one or more of the aromatic epoxide derived compound, the one or more reactants, a third solvent, a third buffer, a third salt, and a third enzyme. In embodiments, the synthesis reaction mixture may further be configured to catalyze the synthesis reaction, especially the third enzyme may catalyze the synthesis reaction. The synthesis reaction mixture may be configured to provide specific reaction conditions, such as a specific pH, temperature, chemical catalyst and/or ionic strength. The term “synthesis reaction mixture” may also refer to a plurality of synthesis reaction mixtures. Especially, in embodiments, the synthesis stage may comprise a plurality of successive reactions, wherein at least two of the successive reactions occur in the presence of two different synthesis reaction mixtures.
In embodiments, the synthesis stage may comprise reacting the aromatic epoxide derived compound and the one or more reactants at about a third temperature. In further embodiments, the third temperature may especially be selected from the range of 0-100 °C, more especially from the range of 25-70 °C.
In a second aspect, the current invention also provides an enantiomerically pure aromatic epoxide derived compound obtainable by the production process as described herein. Especially a substantially enantiomerically pure aromatic epoxide derived compound, such as an aromatic epoxide derived compound having an enantiomeric excess (ee) selected from the range of 0.8-1, such as from the range of 0.9-1, especially from the range of 0.95-1. In embodiments, the aromatic epoxide derived compound may have an ee equal to 1, i.e., the aromatic epoxide derived compound may be enantiomerically pure. Especially, the term enantiomerically pure refers to all molecules in a sample having the same chirality sense, especially the same chirality sense within detection limits.
In embodiments, the (substantially) enantiomerically pure aromatic epoxide derived compound may comprise a product of the synthesis stage, i.e., the (substantially) enantiomerically pure aromatic epoxide derived compound may comprise the second aromatic epoxide derived compound.
In embodiments, the aromatic epoxide derived compound may comprise one or more of (lS,6S)-6-azidocyclohexa-2,4-dien-l-ol, (lS,6S)-6aminocyclohexa-2,4-dien-l-ol, (lR,6R)-6-azidocyclohexa-2,4-dien-l-ol, (lR,6R)-6-aminocyclohexa-2,4-dien-1 -ol, (1R,2R)-1 -azido-1,2-dihydronaphthalen-2-ol, (I R,2R)-1 -amino-1,2-dihydronaphthalen-2-ol, (1 S,2 S)-2-azido-1,2dihydronaphthalen-1 -ol, (1 S,2S)-2-amino-1,2-dihydronaphthalen-1 -ol, (1R,2R)1 -azido-1,2-dihydroanthracen-2-ol, (1R,2R)-1 -azido-1,2-dihydrophenanthren-2ol, and (3R,4S)-4-azido-3,4-dihydrodibenzo[b,d]furan-3-ol.
In embodiments, two or more of the epoxidation stage, the ringopening-stage and the synthesis stage may be temporally and/or spatially separated. Hence, in embodiments the epoxidation stage and the ring-opening stage may be temporally and/or spatially separated. Alternatively or additionally, the ring-opening stage and the synthesis stage may be temporally and/or spatially separated. The temporal and/or spatial separation may be beneficial as suitable, especially optimal, reaction conditions may differ for the different stages. For example, in embodiments the nucleophile reacting in the ring-opening stage may negatively affect the performance of the oxidoreductase in the epoxidation stage.
In embodiments wherein the synthesis stage comprises a plurality of successive reactions, the successive reactions may be spatially and/or temporally separated. Hence, in embodiments, the synthesis reaction mixture may comprise a plurality of synthesis reaction mixtures which are used in succession.
In embodiments, one or more of the epoxidation reaction mixture, the ring-opening reaction mixture and the synthesis reaction mixture may be periodically or continuously adjusted. For example, the pH of the epoxidation (ring-opening / synthesis) reaction mixture may be modified during the process.
In embodiments, the epoxidation stage may comprise supplying the aromatic compound and the peroxide to an enzyme reactor comprising oxidoreductase, wherein the enzyme reactor is configured to provide the aromatic epoxide to a nucleophile reaction unit, and wherein the ring-opening stage comprises reacting the aromatic epoxide and the nucleophile in the nucleophile reaction unit. In a further embodiment, the enzyme reactor may comprise immobilized oxidoreductase.
In further or alternative embodiments, the nucleophile reaction unit may be in fluid connection with an external recovery loop, wherein contents of the nucleophile reaction unit continuously or periodically pass through the external recovery loop, and wherein the external recovery loop is configured to return the aromatic epoxide and the nucleophile to the nucleophile reaction unit and to remove the aromatic epoxide derived compound. In further embodiments, the external recovery loop may comprise an aromatic epoxide derived compound tubing and an external recovery loop reactor. The aromatic epoxide derived compound tubing may be configured to provide a fluid contact between the nucleophile reaction unit and the external recovery loop reactor. The external recovery loop reactor may be configured to enable the in situ removal of the produced aromatic epoxide derived compound, especially to enable the in situ removal of the produced aromatic epoxide derived compound on a solid phase. Additionally or alternatively, the external recovery loop may be configured to return the nucleophile back to the nucleophile reaction unit. Yet additionally or alternatively, the external recovery loop may be configured to return the aromatic epoxide back to the nucleophile reaction unit. Hence, in embodiments, the external recovery loop reactor may be configured to enable the in situ removal of the produced aromatic epoxide derived compound while simultaneously returning the nucleophile and/or the aromatic epoxide back to the nucleophile reaction unit, especially the nucleophile and the aromatic epoxide. In alternative or further embodiments, the external recovery loop reactor may comprise an outlet configured for the release of the aromatic epoxide derived compound.
In yet further or alternative embodiments, the nucleophile reaction unit or the external recovery loop provides the aromatic epoxide derived compound to a synthesis reaction unit, wherein the synthesis stage comprises reacting the aromatic epoxide derived compound and the one or more reactants in the synthesis reaction unit.
In yet further or alternative embodiments, the synthesis reaction unit may be in fluid connection with a third external recovery loop, wherein contents of the synthesis reaction unit continuously or periodically pass through the third external recovery loop, and wherein the third external recovery loop is configured to return the aromatic epoxide derived compound and the one or more reactants to the synthesis reaction unit and to remove the second aromatic epoxide derived compound.
In yet further or alternative embodiments, the enzyme reactor may - during use - comprise a first reaction mixture and/or the nucleophile reaction unit may - during use - comprise a second reaction mixture and/or the synthesis reaction unit may - during use - comprise a third reaction mixture.
In yet further or alternative embodiments, the enzyme reactor may comprise a first inlet configured to supply a component of the first reaction mixture, especially all components of the first reaction mixture. Alternatively or additionally, the nucleophile reaction unit may comprise a second inlet configured to supply a component of the second reaction mixture. Alternatively or additionally, the synthesis reaction unit may comprise a third inlet configured to supply a component of the third reaction mixture.
In further embodiments, the first inlet may be functionally coupled to one or more tubings. For example, the first inlet may be functionally coupled to an aromatic compound inlet tubing supplying the aromatic compound and/or to a peroxide inlet tubing supplying the peroxide. In embodiments, the aromatic compound inlet tubing may be configured to provide a fluid connection between the first inlet and an aromatic compound source. It will be clear to a person skilled in the art that each of the inlets may be functionally coupled to one or more tubings providing one or more components of a reaction mixture.
In further or alternative embodiments, the enzyme reactor and the nucleophile reaction unit may be functionally coupled through an aromatic epoxide tubing. The aromatic epoxide tubing may be configured to provide the aromatic epoxide to the nucleophile reaction unit from the enzyme reactor. In yet further or alternative embodiments, tubing may be arranged between the nucleophile reaction unit and one or more device elements selected from the group comprising the external recovery loop, the external recovery loop reactor, and the synthesis reactor, especially wherein the tubing is configured to provide a fluid connection between the nucleophile reaction unit and the device element. In yet further or alternative embodiments, tubing may be arranged between two synthesis reactors, especially wherein the tubing is configured to provide a fluid connection between the two synthesis reactors.
The production process may be part of or may be applied in e.g. the synthesis of bulk and/or specialty chemicals comprising, for example, triazole compounds, aromatic azides, amino alcohols, and aromatic amines, with applications, for example, as active pharmaceutical ingredients, agrochemicals, pharmaceutical therapeutics, antibiotics, medicine, and fluorescence probes, dyes, and antioxidants, and as substrates for further organic syntheses.
In a third aspect, the invention also provides a production process for producing an aromatic epoxide, the process comprising: reacting an aromatic compound and a peroxide in the presence of an oxidoreductase, resulting in the aromatic epoxide, wherein the aromatic compound comprises an aromatic ring structure, wherein the oxidoreductase catalyzes epoxidation of the aromatic ring structure, and wherein the oxidoreductase is especially selected from the group consisting of oxygenases, peroxygenases, oxidases, and peroxidases. In a fourth aspect, the invention also provides a product obtainable by such production process. The herein described embodiments - as far as related to the epoxidation stage - may also apply to these aspects of the invention.
In a fifth aspect, the invention also provides a production process for producing an aromatic epoxide derived compound, the process comprising: reacting the aromatic epoxide and a nucleophile, resulting in the aromatic epoxide derived compound. In a sixth aspect, the invention also provides a product obtainable by such production process. The herein described embodiments - as far as related to the ring-opening stage - may also apply to these aspects of the invention. Hence, the embodiments described herein are not limited to a single aspect of the invention. For example, an embodiment describing an epoxidation step may be an embodiment of the first aspect as well as of the third aspect, and a product mentioned in relation to such embodiment may further relate to the second, fourth and optionally sixth aspect of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
Embodiments of the invention will now be described, by way of example only, with reference to the accompanying schematic drawings in which corresponding reference symbols indicate corresponding parts, and in which:
Fig. 1 schematically depicts an embodiment of the production process according to the invention wherein naphthalene is the aromatic compound;
Fig. 2 schematically depicts an embodiment of the production process according to the invention, wherein the epoxidation stage and the ringopening stage are spatially separated;
Fig. 3 A-B depict experimental observations regarding the stability the aromatic epoxide produced in the epoxidation stage;
Fig. 4 depicts several aromatic epoxide derived compounds that were obtained through the production process as described herein;
Fig. 5 depicts several further aromatic epoxide derived compounds that may be obtained through the production process as described herein; and
Fig. 6 depicts several non-limiting aromatic epoxide derived compounds that may be obtained using herein described procedures.
The schematic drawings are not necessarily on scale.
DETAILED DESCRIPTION OF THE EMBODIMENTS
Fig. 1 schematically depicts an embodiment of the production process 10 according to the invention. The epoxidation stage 200 comprises reacting the aromatic compound 100 with the peroxide 210 resulting in the aromatic epoxide 110, wherein the aromatic compound 100 comprises an aromatic ring structure, wherein the oxidoreductase 220 catalyzes epoxidation of the aromatic ring structure, and wherein the oxidoreductase 220 is selected from the group consisting of oxygenases, peroxygenases, oxidases, and peroxidases. The ring-opening stage 300 comprises reacting the aromatic epoxide 110 and a nucleophile 310 resulting in the aromatic epoxide derived compound 120. The synthesis stage 400 comprises reacting the aromatic epoxide derived compound 120 with one or more reactants 410 resulting in the formation of a second aromatic epoxide derived compound 120b. In this embodiment, the aromatic compound 100 comprises naphthalene, the peroxide 210 comprises H2O2, the oxidoreductase 220 comprises an unspecific peroxygenase, especially the unspecific peroxygenase Apol of Agrocybe aegenla, the aromatic epoxide 110 comprises naphthalene oxide, the nucleophile 310 comprises azide, the aromatic epoxide derived compound 120 comprises (lR,2R)-l-azido-l,2-dihydronaphthalen-2-ol, the one or more reactants 410 comprise HCIO4, and the second aromatic epoxide derived compound 120b comprises 1-azidonaphthalene. Hence, In this embodiment, wherein the peroxide 210 comprises hydrogen peroxide, hence, H2O may be produced as a by-product of the epoxidation reaction. In other embodiments, wherein the peroxide 210 does not comprise hydrogen peroxide but one or more other peroxides, one or more different by-products may be produced.
In embodiments, the epoxidation stage may comprise reacting the aromatic compound 100 and the peroxide 210 in the presence of a first solvent for the aromatic compound 100. In further embodiments, the first solvent may comprise an aqueous solvent. In alternative embodiments, the first solvent may comprise one or more of a nonpolar solvent, a polar aprotic solvent, or a supercritical solvent.
Fig. 1 further depicts the spontaneous NIH shift 500 converting the aromatic epoxide 110 into the aromatic alcohol 115. The NIH shift 500 is an undesired side reaction with respect to the invention and may be minimized through design of the first reaction mixture and/or the second reaction mixture as well as through minimizing the time between the epoxidation stage 200 and the ring-opening stage 300. For example, in embodiments, the epoxidation stage may comprise reacting the aromatic compound 100 and the peroxide 210 in the presence of a first solvent for the aromatic compound 100, wherein the first solvent is chosen such that the occurrence of the NIH shift 500 is minimized. As FFO may catalyze the NIH shift, the first solvent may, for example, comprise one or more of a nonpolar solvent, a polar aprotic solvent or a supercritical solvent.
Fig. 2 schematically depicts an embodiment of the production process. The epoxidation stage 200 comprises supplying the aromatic compound 100 and the peroxide 210 to an enzyme reactor 250. The enzyme reactor 250 comprises a first reaction mixture 230. The enzyme reactor 250 further comprises a first inlet 260 configured to provide a component of the first reaction mixture 230, herein specifically the aromatic compound 100 via an aromatic compound inlet tubing 102 and the peroxide 210 via a peroxide inlet tubing 212. In embodiments, the first reaction mixture 230 may comprise the oxidoreductase 220. In further or alternative embodiments, the oxidoreductase 220 may be immobilized on a structure in the enzyme reactor 250, i.e., the enzyme reactor 250 may comprise immobilized oxidoreductase. In such embodiment, the aromatic compound 100 and the peroxide 110 may react in the enzyme reactor resulting in the aromatic epoxide 110. The enzyme reactor 250 may be configured to provide the aromatic epoxide 110 to the nucleophile reaction unit 350. In this embodiment, the enzyme reactor 250 is configured to provide the aromatic epoxide 110 to the nucleophile reaction unit 350 via an aromatic epoxide tubing 112. The ring-opening stage 300 comprises reacting the aromatic epoxide 110 and the nucleophile 310 in the nucleophile reaction unit 350 resulting in the aromatic epoxide derived compound 120. The nucleophile reaction unit 350 may comprise a second reaction mixture 330. In this embodiment, the nucleophile reaction unit 350 is in fluid connection with an external recovery loop 370. Hence, contents of the nucleophile reaction unit 350 continuously or periodically pass through the external recovery loop. The external recovery loop 370 may be configured to return the aromatic epoxide 110 and the nucleophile 310 to the nucleophile reaction unit 350 and to remove the aromatic epoxide derived compound 120. In this embodiment, the external recovery loop 370 comprises an aromatic epoxide derived compound tubing 122 and a external recovery loop reactor 380. The aromatic epoxide derived compound tubing 122 is configured to provide a fluid contact between the nucleophile reaction unit 350 and the external recovery loop reactor 380. The external recovery loop reactor 380 may be configured to enable the in situ removal of the produced aromatic epoxide derived compound on a solid phase while simultaneously returning the nucleophile back to the nucleophile reaction unit. In alternative or further embodiments, the external recovery loop reactor 380 may comprise an outlet configured for the release of the aromatic epoxide derived compound. In the depicted embodiment, the epoxidation stage and the ring-opening stage are spatially separated. In an alternative embodiment, the epoxidation stage and the ring-opening stage may be temporally separated.
Hereinafter several non-limiting experimental procedures are provided which are each either an embodiment of at least part of the production process as described herein, or an analytical procedure to quantify an aromatic epoxide and/or an aromatic epoxide derived compound. It will be clear to a person skilled in the art that many modifications will be possible to these procedures while remaining in the scope of the present invention.
Procedure 1: Enzymatic epoxidation reactions to prepare aromatic epoxides. 2 mM naphthalene, 2 mM H2O2 and 200 nM unspecific peroxygenase of Agrocybe aegerita were added into 1.0 mL NaPi pH 7 (with 20% MeCN). The mixture was slowly stirred for 2.5-15 min, and then the mixture was moved to an ice bath and kept stirring for 30 min. The above concentration of each component is the final concentration under the condition applied. The mixture was then extracted with DCM and dried over MgSO4 at 0 °C. Finally, the solvent was evaporated at 0 °C and the product was purified by column using 10 % ethyl acetate in either pentane or heptane. For the extraction, diethyl ether, chloroform or ethyl acetate can also be used. The amount of co-solvent MeCN can vary from 0-70 % v/v in NaPi buffer. Other co-solvents such as methanol, acetone, ethanol, dimethyl sulfoxide are also applicable. The epoxidation may be performed at a temperature selected from the range of 0 - 45 °C. The pH may be selected from the range of 5.0 to 8.5. In alternative embodiments, other buffers may be used in the reactions such as a buffer selected from the group comprising sodium citrate, sodium acetate, Tris-H2SO4, Bis-Tris, BES buffer and britton-robinson buffer, with the buffer concentration selected from the range of 10 to 500 mM.
Procedure 2: Quantification of naphthalene epoxide. A reaction mixture with 100-400 nM unspecific peroxygenase of Agrocybe aegerita, 2 mM naphthalene and 2 mM H2O2 in NaPi (pH 7.0 with 20% CH3CN) was firstly prepared, then 20 pL of this mixture was added and mixed quickly to a 1980 pL of NaPi (pH 7.0 with 20% CH3CN). The final concentration of naphthalene for UV assay was 0.02 mM. The absorbance at 266.5 nm (extinction coefficient 8850 mM^cnf1) was recorded and the corresponding naphthalene epoxide concentration is shown in Fig. 3a. Fig. 3a depicts the concentration of naphthalene epoxide C over time τ in minutes with 100 nM unspecific peroxygenase (line Ai), with 200 nM unspecific peroxygenase (line A2), and with 400 11M unspecific peroxygenase (line A3).
Procedure 3: Quantification of naphthalene epoxide. A reaction mixture with 200 nM unspecific peroxygenase of Agrocybe aegerita, 2 mM naphthalene and 1-4 mM H2O2 in NaPi (pH 7.0 with 20% CH3CN) was firstly prepared, then 20 pL of this mixture was added and mixed quickly to a 1980 pL of NaPi (pH 7.0 with 20% CH3CN. The absorbance of this mixture at 266.5 nm (extinction coefficient 8850 mM^cm'1) was recorded and the corresponding naphthalene epoxide shown in Fig. 3b. Fig. 3b depicts the concentration of naphthalene epoxide C over time τ in minutes with 1 mM H2O2 (line A4), with 2 mM H2O2 (line A5), and with 4 mM H2O2 (line As). It is clear from Fig. 3a-b that naphthalene epoxide is present for several minutes under these conditions rather than that it immediately undergoes the NIH shift.
Procedure 4: Ring-opening reactions with nucleophiles. 2 mM naphthalene, 2 mM H2O2 and 200 nM unspecific peroxygenase of Agrocybe aegerita were added into 1.0 mL NaPi pH 7 (with 20% MeCN). The mixture was slowly stirred for 2.5-10 min, and then the nucleophiles (e.g. N?,-, CN-) with a certain concentration (0.01 to 0.25 M) were added. In other embodiments, the mixture may be slowly stirred for more than 10 minutes, such as for 15 minutes, especially 25 minutes. The above concentration of each component is the final concentration under the condition applied. The mixture was then extracted with DCM and dried over MgSCfi. Finally, the solvent was evaporated at 30 °C and the aromatic epoxide derived compound was purified by column using 10-20 % ethyl acetate in pentane/heptane. Other co-solvents, buffer and reaction conditions such as described in procedure 1 “Enzymatic epoxidation reactions to prepare aromatic epoxides” may also be used here. The concentration of the aromatic epoxide derived compound was quantified using high-performance liquid chromatography (HPLC) and is reported in the table below. The indicated conversion refers to the percentage of naphthalene converted to the aromatic epoxide derived compound according to the combination of procedures 1 and 4.
N3- [M] Reaction time [min] Conversion [%]
0.25 2.5 68
0.125 2.5 74
0.0625 2.5 48.5
0.03125 2.5 30.5
0.125 3 71
0.125 4 72.5
0.125 6 67
0.125 10 63.5
In embodiments, the nucleophile may comprise one or more of n3; cn; scn-, ocn-, s2; roh, ro; ci-, Br-, ι-, hco2; no2; ch3co2; CH3COS', (CH3CH2)3N, (CH3CH2)3P, HRs, H2S, RNH2, RNHR’, R-SH, PhSH, PhSeH, or PhOH.
Hence, in a specific embodiment of the production process, the aromatic compound comprises naphthalene, the oxidoreductase comprises the Apol protein (unspecific peroxygenase) of Agrocybe aegerita, and the nucleophile comprises N3’.
Procedure 5: Elimination of H2O to aromatic azides. In a one-pot three-step manner, after the epoxidation and ring-opening of napthalene using nucleophiles (Procedure 4 “Ring-opening reactions with nucleophiles”), perchloric acid (0.5-3 M) was added to the reaction mixture and the reaction mixture was stirred for 5 hours. In alternative embodiments, trifluoroacetic acid (0.5-3 M) may be used instead of perchloric acid. The extraction and purification procedures are the same as mentioned in procedure 1 “Enzymatic epoxidation reactions to prepare epoxides”. The concentration of the second aromatic epoxide derived compound was quantified using gas chromatography (GC) and is reported in the table below. The indicated conversion refers to the percentage of naphthalene converted to the second aromatic epoxide derived compound according to the combination of procedures 1,4 and 5.
NaN3 [M] HC1O4 [M] Conversion (%)
0.25 3 44.8
0.1 3 88.3
0.05 3 59
0.01 3 27
0.25 1 42.3
0.25 0.5 -
Procedure 6: Reduction of aromatic azide to aromatic amine. The reaction was performed in a one-pot three-step manner. After the epoxidation and ring-opening reaction with nucleophiles, the pH of the solution was then adjusted to approx. 9 by using NaOH (5 M). An amount of 10 mol% Pd/C was added directly to the solution. N2 gas was bubbled through the reaction mixture for 10 min, the reaction vial was sealed, and a balloon of pure H2 was applied to initiate the reduction. After 6 hours, the second aromatic epoxide derived product was extracted with DCM. The organic phase was dried over MgSO4 and the solvent DCM was slowly evaporated in a rotavap. The second aromatic epoxide derived product was purified by column using 10% ethyl acetate in pentane. The extraction and purification procedures are the same as above mentioned in procedure 1 “enzymatic epoxidation reactions to prepare aromatic epoxides”. A full conversion of aromatic azide to aromatic amine was observed using quantification via GC-MS.
Procedure 7: Reduction of azido alcohol to amino alcohols. The reaction was performed in a one-pot three-step manner in a preparative scale. In a 100 ml reaction scale, 2 mM of naphthalene, 2 mM of H2O2 and 200 nM of unspecific peroxygenase were applied. After the addition of NaNs, the reaction was continued for 4 hours. The pH of the solution was then adjusted to approx. 9 by using NaOH (5 M). An amount of 10 mol% Pd/C was added directly to the solution. N2 gas was bubbled through the reaction mixture for 10 min, the reaction vial was sealed, and a balloon of pure H2 was applied to initiate the reduction. After 6 hours, the product was extracted with DCM. The organic phase was dried over MgSO4 and the solvent DCM was slowly evaporated in a rotavap. The product was purified by column using 10% ethyl acetate in pentane. The extraction and purification procedures are the same as above mentioned in procedure 1 “enzymatic epoxidation reactions to prepare aromatic epoxides”. A full conversion of azido alcohol to amino alcohol was observed using quantification via NMR.
Procedure 8: Click chemistry. The reactions were performed in a two-pot three-step manner; however, in embodiments the reactions may also be performed in a sequential one-pot three-step manner. As to the reaction in twopot three-step manner, the azido alcohol and alkyne (e.g. 1.5 eq. phenyl acetylene) were dissolved in H2O and tert-butyl alcohol (2:1, v:v). 5mol %
CuSO4*5H2O and 10mol% sodium ascorbate were added. The reaction mixture was stirred for 8 hours at 30 °C. The product was extracted with DCM. The organic phase was dried over MgSCE and the solvent DCM was slowly evaporated in a rotavap. The extraction and purification procedures are the same as above mentioned in “Enzymatic epoxidation reactions to prepare aromatic epoxides”. A 41% conversion of azido alcohol was observed using quantification via 1H NMR using phenyl acetylene as alkyne. In a separate experiment, a 73% conversion of azido alcohol was observed using quantification via 1H NMR based on propargyl acetate as alkyne.
Aforementioned experimental procedures represent non-limiting embodiments for the epoxidation stage (procedure 1), for the ring-opening stage (procedure 4), and for the synthesis stage (procedures 5-8). It will be clear to one skilled in the art that the synthesis stage may comprise one or more of the aforementioned procedures and/or one or more other procedures not explicitly mentioned herein.
The aforementioned analytical procedures involving spectroscopy, HPLC, GC GC-MS, NMR, 1H NMR are commonly used analytical procedures and will be known to one skilled in the art.
Fig. 4 schematically depicts several aromatic epoxide derived compounds (P15-P33) that were obtained using procedures 1,4 with azide as nucleophile and varying aromatic compounds. The percentages indicate the conversion of the aromatic compound to the aromatic epoxide derived compound using the native unspecific peroxygenase from Agrocybe aegerita (the left percentage) or the “Solo-variant” of the unspecific peroxygenase (the right percentage). “ND” refers to not determined, i.e., not tested. refers to no observed conversion.
Fig. 5 schematically depicts several compounds that were obtained using procedures 1,4 with naphthalene or 1-chloronapthalene as aromatic compound and varying nucleophiles. The percentages indicate the conversion of the aromatic compound to the aromatic epoxide derived compound using the native unspecific peroxygenase from Agrocybe aegerita. The nucleophiles and compounds are: HCO2' to produce l-hydroxy-l,2-dihydro naphthal en-2-yl formate (Nf), CN' to produce 1-hydroxy-1,2-dihydronaphthalene-2-carbonitrile (N2), NO3 to produce 1-hydroxy-1,2-dihydronaphthalen-2-yl nitrate (N3), OCN' to produce 2-(isocyanooxy)-l,2-dihydronaphthalen-l-ol (N4), SCN' to produce 2-thiocyanato-l,2-dihydronaphthalen-l-ol (N5), PhNCS to produce 2-(((phenyl-13-sulfaneylidene)methylene)amino)-l,2dihydronaphthalen-l-ol (N6), HCO2- to produce 5-chl oro-1-hydroxy-1,2dihydronaphthalen-2-yl formate (N7) (from 1-chloronapthalene).
Fig. 6 schematically depicts several non-limiting aromatic epoxide derived compounds that may be obtained using the aforementioned procedures. In embodiments, the depicted compounds may be the final product of the production process. In alternative embodiments, the depicted compounds may be subjected to one or more further reactions in the synthesis stage. Examples of the aromatic epoxide derived compounds include: (TS,6S)-6-azidocyclohexa-2,4dien-l-ol (Pl), (lS,6S)-6-aminocyclohexa-2,4-dien-l-ol (P2), azidobenzene (P3), (lR,6R)-6-azidocyclohexa-2,4-dien-l-ol (P4), (lR,6R)-6-aminocyclohexa-2,4dien-l-ol (PS), (lR,2R)-l-azido-l,2-dihydronaphthalen-2-ol (P6), (1R,2R)-
1- amino-l,2-dihydronaphthalen-2-ol (P7),4-azidonapthalene (P8), (1S,2S)-
2- azido-5-bromo-l,2-dihydronaphthalen-l-ol (P9), (lS,2S)-2-amino-5-bromo1,2-dihydronaphthalen-l-ol (PIO), l-azido-5-bromonapthalene (Pl 1), (1R,2R)-1azido-l,2-dihydroanthracen-2-ol (P12), (3R,4S)-4-azido-3,4-dihydrodibenzo[b,d]furan-3-ol (P13), (lR,2R)-l-azido-l,2-dihydrophenanthren-2-ol (Pl4), and one or more derivatives of one or more of the aforementioned aromatic epoxide derived compounds. In a derivative of an aforementioned aromatic epoxide derived compound one or more of the H groups may be independently replaced with a side group selected from the group comprising F, Cl, Br, I, CF3, CH3, C2H5, NO2, C=N, and (j, JE.2X(). Especially, one or more of the H side groups of the aromatic ring structure may be independently replaced with a side group selected from the group comprising F, Cl, Br, I, CF3, CH3, C2H5, NO2, C=N, and Ci JE 2\O
For example, non-limiting embodiments of azidobenzene derivatives include l-azido-4-methylbenzene and l-azido-2-chloro-4fluorobenzene.
The term “plurality” refers to two or more.
The terms “substantially” or “essentially” herein, and similar terms, will be understood by the person skilled in the art. The terms “substantially” or “essentially” may also include embodiments with “entirely”, “completely”, “all”, etc. Hence, in embodiments the adjective substantially or essentially may also be removed. Where applicable, the term “substantially” or the term “essentially” may also relate to 90% or higher, such as 95% or higher, especially 99% or higher, even more especially 99.5% or higher, including 100%.
The term “comprise” includes also embodiments wherein the term “comprises” means “consists of’.
The term “and/or” especially relates to one or more of the items mentioned before and after “and/or”. For instance, a phrase “item 1 and/or item 2” and similar phrases may relate to one or more of item 1 and item 2. The term comprising may in an embodiment refer to consisting of' but may in another embodiment also refer to containing at least the defined species and optionally one or more other species.
Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.
The devices, apparatus, or systems may herein amongst others be described during operation. As will be clear to the person skilled in the art, the invention is not limited to methods of operation, or devices, apparatus, or systems in operation.
It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design many alternative embodiments without departing from the scope of the appended claims.
In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.
Use of the verb to comprise and its conjugations does not exclude the presence of elements or steps other than those stated in a claim. Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.
The article a or an preceding an element does not exclude the presence of a plurality of such elements.
The invention may be implemented by means of hardware comprising several distinct elements, and by means of a suitably programmed computer. In a device claim, or an apparatus claim, or a system claim, enumerating several means, several of these means may be embodied by one and the same item of hardware. The mere fact that certain measures are recited in mutually different dependent claims does not indicate that a combination of these measures cannot be used to advantage.
The invention also provides a control system that may control the device, apparatus, or system, or that may execute the herein described method or process. Yet further, the invention also provides a computer program product, when running on a computer which is functionally coupled to or comprised by the device, apparatus, or system, controls one or more controllable elements of such device, apparatus, or system.
The invention further applies to a device, apparatus, or system comprising one or more of the characterizing features described in the description and/or shown in the attached drawings. The invention further pertains to a method or process comprising one or more of the characterizing features described in the description and/or shown in the attached drawings.
The various aspects discussed in this patent can be combined in order to provide additional advantages. Further, the person skilled in the art will understand that embodiments can be combined, and that also more than two embodiments can be combined. Furthermore, some of the features can form the basis for one or more divisional applications.

Claims (17)

ConclusiesConclusions 1. Een productieproces voor de productie van een verbinding afgeleid van een aromatische epoxide, waarbij het productieproces omvat:A production process for the production of a compound derived from an aromatic epoxide, the production process comprising: (i) een epoxidatiestadium omvattende: het laten reageren van een aromatische verbinding en een peroxide in de aanwezigheid van een oxidoreductase, resulterend in een aromatische epoxide, waarbij de aromatische verbinding een aromatische ringstructuur omvat, waarbij het oxidoreductase epoxidatie van de aromatische ringstructuur katalyseert, en waarbij het oxidoreductase wordt gekozen uit de groep bestaande uit oxygenasen, peroxygenasen, oxidasen en peroxidasen;(i) an epoxidation stage comprising: reacting an aromatic compound and a peroxide in the presence of an oxidoreductase, resulting in an aromatic epoxide, wherein the aromatic compound comprises an aromatic ring structure, wherein the oxidoreductase catalyzes epoxidation of the aromatic ring structure, and wherein the oxidoreductase is selected from the group consisting of oxygenases, peroxygenases, oxidases and peroxidases; (ii) een ringopeningsstadium omvattende: het reageren van het aromatische epoxide en een nucleofiel deeltje, resulterend in de verbinding afgeleid van een aromatische epoxide.(ii) a ring opening stage comprising: reacting the aromatic epoxide and a nucleophilic particle, resulting in the compound derived from an aromatic epoxide. 2. Het productieproces volgens één van de voorgaande conclusies, waarbij het epoxidatiestadium het laten reageren van de aromatische verbinding en het peroxide in de aanwezigheid van een eerste oplosmiddel voor de aromatische verbinding omvat.The production process according to any of the preceding claims, wherein the epoxidation stage comprises reacting the aromatic compound and the peroxide in the presence of a first solvent for the aromatic compound. 3. Het productieproces volgens conclusie 2, waarbij het eerste oplosmiddel een waterig oplosmiddel omvat.The production process of claim 2, wherein the first solvent comprises an aqueous solvent. 4. Het productieproces volgens één van de voorgaande conclusies, waarbij het epoxidatiestadium het laten reageren van de aromatische verbinding en het peroxide bij een pH gekozen uit het bereik van 5,0-8,5 omvat.The production process according to any of the preceding claims, wherein the epoxidation stage comprises reacting the aromatic compound and the peroxide at a pH selected from the range of 5.0-8.5. 5. Het productieproces volgens conclusie 2, waarbij het eerste oplosmiddel één of meer van een niet-polair oplosmiddel, een polair aprotisch oplosmiddel of een superkritisch oplosmiddel omvat.The production process of claim 2, wherein the first solvent comprises one or more of a non-polar solvent, a polar aprotic solvent, or a supercritical solvent. 6. Het productieproces volgens één van de voorgaande conclusies, waarbij het productieproces verder omvat:The production process according to any of the preceding claims, wherein the production process further comprises: (iii) een synthesestadium omvattende: het laten reageren van de verbinding afgeleid van een aromatische epoxide met één of meer reactanten, resulterend in een tweede verbinding afgeleid van een aromatische epoxide.(iii) a synthesis stage comprising: reacting the compound derived from an aromatic epoxide with one or more reactants, resulting in a second compound derived from an aromatic epoxide. 7. Het productieproces volgens één van de voorgaande conclusies, waarbij het oxidoreductase een niet-specifieke peroxygenase omvat.The production process according to any of the preceding claims, wherein the oxidoreductase comprises a non-specific peroxygenase. 8. Het productieproces volgens één van de voorgaande conclusies, waarbij het nucleofiele deeltje één of meer van N3', CN', SCN-, OCN-, S2‘, ROH, RO', Cl', Br', Γ, HCO2; NO/, CH3CO2', CH3COS', (CH3CH2)3N, (CH3CH2)3P, nh3, H2S, RNH2, RNHR, R-SH, PhSH, PhSeH of PhOH omvat.The production process according to any of the preceding claims, wherein the nucleophilic particle is one or more of N 3 ', CN', SCN, OCN, S 2 ', ROH, RO', Cl ', Br', Γ, HCO 2 ; NO /, CH 3 CO 2 ', CH 3 COS', (CH 3 CH 2 ) 3 N, (CH 3 CH 2 ) 3 P, nh 3 , H 2 S, RNH 2 , RNHR, R-SH, PhSH, PhSeH or PhOH. 9. Het productieproces volgens één van de voorgaande conclusies, waarbij het epoxidatiestadium en het ringopeningsstadium temporeel en/of ruimtelijk gescheiden zijn.The production process according to any of the preceding claims, wherein the epoxidation stage and the ring opening stage are temporally and / or spatially separated. 10. Het productieproces volgens conclusie 9, waarbij het epoxidatiestadium het toevoeren van de aromatische verbinding en het peroxide aan een enzymreactor omvattende geïmmobiliseerd oxidoreductase omvat, waarbij de enzymreactor geconfigureerd is om het aromatische epoxide aan een nucleofielereactie-eenheid te verschaffen, en waarbij het ringopeningsstadium het laten reageren van het aromatische epoxide en het nucleofiele deeltje in de nucleofielereactie-eenheid omvat.The production process of claim 9, wherein the epoxidation stage comprises supplying the aromatic compound and peroxide to an enzyme reactor comprising immobilized oxidoreductase, wherein the enzyme reactor is configured to provide the aromatic epoxide to a nucleophilic reaction unit, and wherein the ring opening stage is the reacting the aromatic epoxide and comprising the nucleophilic particle in the nucleophilic reaction unit. 11. Het productieproces volgens conclusie 10, waarbij de nucleofielereactieeenheid in fluïde-verbinding staat met een externe terugwinningsomloop, waarbij inhoud van de nucleofielereactie-eenheid continu of periodiek door de externe terugwinningsomloop passeert en waarbij de externe terugwinningsomloop geconfigureerd is om het aromatische epoxide en het nucleofiele deeltje naar de nucleofielereactie-eenheid terug te brengen en om verbinding afgeleid van een aromatische epoxide weg te halen.The production process according to claim 10, wherein the nucleophilic reaction unit is in fluid communication with an external recovery circulation, wherein content of the nucleophilic reaction unit passes continuously or periodically through the external recovery circulation and wherein the external recovery circulation is configured around the aromatic epoxide and the nucleophilic to return the particle to the nucleophilic reaction unit and to remove compound derived from an aromatic epoxide. 12. Het productieproces volgens een van de conclusies 9-11, waarbij de aromatische verbinding naftaleen omvat, waarbij het oxidoreductase het Apoleiwit van Agrocybe aegerita omvat, en waarbij het nucleofiele deeltje N3' omvat.The production process according to any of claims 9-11, wherein the aromatic compound comprises naphthalene, wherein the oxidoreductase comprises the Apole protein of Agrocybe aegerita, and wherein the nucleophilic particle comprises N 3 '. 13. Het productieproces volgens één van de voorgaande conclusies, waarbij het epoxidatiestadium het laten reageren van de aromatische verbinding en het peroxide bij een temperatuur gekozen uit het bereik van 0-45 °C omvat.The production process according to any of the preceding claims, wherein the epoxidation stage comprises reacting the aromatic compound and the peroxide at a temperature selected from the range of 0-45 ° C. 14. Het productieproces volgens één van de voorgaande conclusies, waarbij de aromatische ringstructuur bestaat uit koolstofatomen.The production process according to any of the preceding claims, wherein the aromatic ring structure consists of carbon atoms. 15. Het productieproces volgens één van de voorgaande conclusies 1-13, waarbij de aromatische ringstructuur een heterocyclische ringstructuur omvat.The production process according to any of the preceding claims 1-13, wherein the aromatic ring structure comprises a heterocyclic ring structure. 16. Een enantiomeer zuivere verbinding afgeleid van een aromatische epoxide verkrijgbaar door het productieproces volgens één van de voorgaande conclusies.An enantiomerically pure compound derived from an aromatic epoxide obtainable by the production process according to any of the preceding claims. 17. De verbinding afgeleid van een aromatische epoxide volgens conclusie 16, waarbij de verbinding afgeleid van een aromatische epoxide één of meer van (lS,6S)-6-azidocyclohexa-2,4-dieen-l-ol, (lS,6S)-6-aminocyclohexa-2,4-dieenl-ol, azidobenzene, (lR,6R)-6-azidocyclohexa-2,4-dieen-l-ol, (lR,6R)-6-aminocyclohexa-2,4-dieen-l-ol , (lR,2R)-l-azido-l,2-dihydronaftaleen-2-ol, (1R,2R)1 -amino-1,2-dihydronaftaleen-2-ol, 4-azidonaftaleen, (1 S,2S)-2-azido-1,2dihydronaftaleen-1 -ol, (1 S,2S)-2-amino-1,2-dihydronaftaleen-1 -ol, 1 -azido-5broom-naftaleen, (1R,2R)-1 -azido-1,2-dihydroanthraceen-2-ol, (1R,2R)-1 -azidol,2-dihydrofenanthren-2-ol en (3R,4S)-4-azido-3,4-dihydro-di-benzo[b,d]-furan3-ol, en één of meer derivaten van één of meer van de hiervoor genoemde verbindingen afgeleid van een aromatische epoxide omvat.The compound derived from an aromatic epoxide according to claim 16, wherein the compound derived from an aromatic epoxide one or more of (1S, 6S) -6-azidocyclohexa-2,4-diene-1-ol, (1S, 6S) -6-aminocyclohexa-2,4-dien-1-ol, azidobenzene, (1R, 6R) -6-azidocyclohexa-2,4-diene-1-ol, (1R, 6R) -6-aminocyclohexa-2,4-diene -1-ol, (1R, 2R) -1-azido-1,2-dihydronaphthalene-2-ol, (1R, 2R) 1 -amino-1,2-dihydronaphthalene-2-ol, 4-azidonaphthalene, (1 S, 2S) -2-azido-1,2-dihydronaphthalene-1-ol, (1 S, 2S) -2-amino-1,2-dihydronaphthalene-1-ol, 1-azido-5-bromo-naphthalene, (1R, 2R ) -1-azido-1,2-dihydroanthracen-2-ol, (1R, 2R) -1-azidol, 2-dihydrophenanthren-2-ol and (3R, 4S) -4-azido-3,4-dihydro- di-benzo [b, d] furan-3-ol, and one or more derivatives of one or more of the aforementioned compounds derived from an aromatic epoxide.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102007016139A1 (en) * 2007-03-30 2008-10-02 Jenabios Gmbh Method for regioselective oxygenation of N-heterocycles

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61109784A (en) 1984-11-02 1986-05-28 Nippon Shokubai Kagaku Kogyo Co Ltd Production of arene oxide
WO2017081355A1 (en) 2015-11-13 2017-05-18 Consejo Superior De Investigaciones Cientificas (Csic) Mutants of unspecific peroxygenase with high monooxygenase activity and uses thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102007016139A1 (en) * 2007-03-30 2008-10-02 Jenabios Gmbh Method for regioselective oxygenation of N-heterocycles

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
ALEXANDER KARICH ET AL: "Benzene oxygenation and oxidation by the peroxygenase of Agrocybe aegerita", AMB EXPRESS, 1 December 2013 (2013-12-01), Berlin/Heidelberg, pages 1 - 8, XP055561517, Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3565944/pdf/2191-0855-3-5.pdf> [retrieved on 20190225], DOI: 10.1186/2191-0855-3-5 *
DAVID AMANTINI ET AL: "Nucleophilic ring opening of 1,2-epoxides in aqueous medium", ARKIVOC, vol. 2002, no. 11, 8 March 2003 (2003-03-08), US, pages 293 - 311, XP055561519, ISSN: 1424-6376, DOI: 10.3998/ark.5550190.0003.b26 *
H. H. KUO ET AL: "Indole peroxygenase activity of indoleamine 2,3-dioxygenase", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 109, no. 35, 13 August 2012 (2012-08-13), US, pages 13966 - 13971, XP055561804, ISSN: 0027-8424, DOI: 10.1073/pnas.1207191109 *
MARTIN KLUGE ET AL: "Hydroxylation of naphthalene by aromatic peroxygenase from Agrocybe aegerita proceeds via oxygen transfer from H2O2 and intermediary epoxidation", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 81, no. 6, 25 September 2008 (2008-09-25), DE, pages 1071 - 1076, XP055561509, ISSN: 0175-7598, DOI: 10.1007/s00253-008-1704-y *
SHELDON W. MAY: "Enzymatic epoxidation reactions", ENZYME AND MICROBIAL TECHNOLOGY, vol. 1, no. 1, 1 January 1979 (1979-01-01), US, pages 15 - 22, XP055561515, ISSN: 0141-0229, DOI: 10.1016/0141-0229(79)90005-X *
SHIV K. AGARWAL ET AL: "Synthesis of arene oxide and trans-dihydrodiol metabolites of quinoline", JOURNAL OF THE CHEMICAL SOCIETY, PERKIN TRANSACTIONS 1, vol. 1, no. 7, 1 January 1990 (1990-01-01), GB, pages 1969 - 1974, XP055561650, ISSN: 0300-922X, DOI: 10.1039/p19900001969 *

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