Preparation process of chiral pyrrotriazole alcohols

Abstract

The present invention relates to formula (I) TIFF2026521749000123.tif33161 This invention relates to a novel process for the preparation of chiral pyrrotriazole alcohols, where X is a halogen atom and n is an integer of 1, 2, or 3, and a helical bond (II) TIFF2026521749000124.tif14161 (III) TIFF2026521749000125.tif17161 or (IV) TIFF2026521749000126.tif14161 Or it represents a mixture of these enantiomers. The chiral pyrrotriazole alcohol of formula (I) is a versatile intermediate for preparing compounds that may function as active pharmaceutical ingredients in drugs.

Description

[Technical Field]

[0001] This invention relates to chiral pyrrotriazole alcohol of formula I. [ka] This relates to a novel process for the preparation of In the formula, X is a halogen atom, and n is an integer of 1, 2, or 3, and the helical bond " [ka] "teeth," [ka] "or" [ka] This represents either a compound of these enantiomers or a mixture thereof. [Background technology]

[0002] Chiral pyrrotriazole alcohol of formula I is a versatile intermediate for preparing compounds that may function as active pharmaceutical ingredients in drugs. For example, chiral pyrrotriazole alcohol of formula I can be used as an intermediate for preparing compounds that may act as gamma-secretase modulators, as disclosed in International Publication No. 2020 / 120521; compounds that may act as melanocortin 4 receptor antagonists, as disclosed in International Publication No. 2021250541; or compounds that may act as ubiquitin-specific processing protease 1, as disclosed in International Publication No. 2021 / 247606.

[0003] The present invention further relates to a novel chiral pyrrolotriazole alcohol of formula I. [ka] (In the formula, X is a halogen atom, n is an integer of 1, 2, or 3, and the helical bond " [ka] "teeth," [ka] "or" [ka] (or a mixture of these enantiomers) [Overview of the project]

[0004] The objective of the present invention was to find a suitable process for preparing this versatile chiral pyrrorotriazole alcohol compound of formula I.

[0005] This objective can be achieved through the process outlined below.

[0006] Chiral pyrrotriazole alcohol of formula I [ka] Process for preparation (In the formula, X is a halogen atom, n is an integer of 1, 2, or 3, and the helical bond " [ka] "teeth," [ka] "or" [ka] (or a mixture of these enantiomers) This is the ketone of formula V. [ka] (wherein X and n are as above) a) Metal complex catalysts in the presence of a reducing agent, or b) Oxidoreductase This includes reduction to form a chiral alcohol of formula I. [Modes for carrying out the invention]

[0007] The following definitions are provided to illustrate and define the meanings and scopes of various terms used in this specification to describe the present invention.

[0008] "C 1-6 The term "-alkyl" refers to a branched or linear monovalent saturated aliphatic hydrocarbon radical consisting of 1 to 6 carbon atoms, preferably 1 to 4, more preferably 1 to 2 carbon atoms. This term is further exemplified by radicals such as methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl or t-butyl, pentyl and its isomers, or hexyl and its isomers.

[0009] "C 1-6 The term "alkoxy" refers to the C defined above, which has an oxygen atom bonded to it. 1-6 - Refers to an alkyl group.

[0010] The term "halogen" refers to fluorine, chlorine, bromine, or iodine, but more specifically to chlorine and bromine.

[0011] The term "aryl" relates to aromatic carbon rings, such as phenyl or naphthyl rings, preferably phenyl rings.

[0012] The term "heteroaryl" refers to aromatic 5-6 member monocyclic or 9-10 member bicyclic rings that may contain one, two, or three heteroatoms selected from nitrogen, oxygen, and / or sulfur, such as pyridinyl, pyrazolyl, pyrimidinyl, benzimidazolyl, quinolinyl, and isoquinolinyl.

[0013] Spiral connection [ka] "teeth" [ka] "or" [ka] It represents the chirality of the molecule, and therefore indicates the chirality of the molecule, but it also represents a mixture of these enantiomers.

[0014] Whenever a chiral carbon is present in a chemical structure, all stereoisomers associated with that chiral carbon are intended to be included in the structure as pure stereoisomers and mixtures thereof.

[0015] The process of the present invention can be illustrated using the following scheme 1. Scheme 1 [ka] In the formula, X is a halogen atom, and n is an integer of 1, 2, or 3, and the helical bond " [ka] "teeth," [ka] "or" [ka] This represents either a compound of these enantiomers or a mixture thereof.

[0016] The reduction is, a) Metal complex catalysts in the presence of a reducing agent, or b) This can be carried out using an oxidoreductase enzyme.

[0017] The ketone of formula V can be prepared according to schemes 2 and 3 below. Scheme 2 [ka]

[0018] This process involves, in the first step, a) 3,5-dihalogen-1H-1,2,4-triazole of formula II [ka] (In the formula, X is a halogen atom) a 1 ) Michael addition using acrylic ester IIIa [ka] (In the formula, R is C 1-4 (It is alkyl) by, or a 2 Alkylation of the halogenated carboxylic acid alkyl ester of formula IIIb [ka] (In the formula, R is C 1-4 (It is an alkyl group, where n is an integer of 1, 2, or 3, and X is a halogen.) Formula IV 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester [ka] (where R, n and X are as defined above) and in the second step, b) ring closure of the 3,5-dihalo-1,2,4-triazole-carboxylic acid ester of formula IV with an organometallic reagent, and

[0019] The 3,5-dihalo-1H-1,2,4-triazole starting compound, preferably 3,5-dibromo-1H-1,2,4-triazole, is commercially available.

[0020] The Michael addition using acrylic ester IIIa and the alkylation with the alkyl halocarboxylate of formula IIIb are typically carried out in the presence of a base selected from organic bases such as triethylamine, N,N-diisopropylethylamine, tributylamine, and inorganic bases such as potassium carbonate or cesium carbonate.

[0021] Triethylamine is the preferred base for the Michael addition using acrylic ester IIIa, and potassium carbonate is the preferred base for the alkylation with the alkyl halocarboxylate of formula IIIb. 5 7> Suitable acrylic esters IIIa are C 1-4 alkyl esters, preferably methyl esters, and suitable alkyl halocarboxylates of formula IIIb are bromo-C 1-4 alkyl esters, preferably methyl esters or ethyl esters.

[0023] The reaction can be carried out in a suitable solvent selected from alcoholic solvents (e.g., methanol, ethanol, isopropanol) or polar aprotic solvents (e.g., DMSO, acetonitrile, THF, MeTHF) at a reaction temperature of 30 °C to 170 °C, preferably in MeTHF at 70 °C.

[0024] N-alkylation of N-containing heterocycles has been reported in the literature (Gmach J. et. al., Synthesis, 2016, 48, 2681-2704).

[0025] Formula IV 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester [ka] (In the formula, R is C 1-4 A (alkyl, where n is an integer of 1, 2, or 3, and X is a halogen) is a novel compound and thus constitutes a further embodiment of the present invention.

[0026] In a preferred embodiment, X is bromine.

[0027] In a more preferred embodiment, R is methyl or ethyl.

[0028] Particularly preferred 3,5-dihalogen 1,2,4-triazole-carboxylic acid esters of formula IV are, X = bromine, R = methyl, and n is 1. X = bromine, R = ethyl, and n is 1. X = bromine, R = methyl, and n is 2. X = bromine, R = ethyl, and n = 2.

[0029] In step b), the 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester of formula IV can be subjected to a ring-closing reaction with an organometallic reagent such as an organolithium or organomagnesium compound.

[0030] Suitable organolithium compounds are n-hexyllithium, n-butyllithium, phenyllithium, or methyllithium.

[0031] Suitable organomagnesium compounds include methylmagnesium bromide, ethylmagnesium chloride, ethylmagnesium bromide, isopropylmagnesium bromide, or isopropylmagnesium chloride.

[0032] The reaction can be carried out in a suitable solvent selected from 2-methyltetrahydrofuran, tetrahydrofuran, toluene, or methyl t-butyl ether, at a reaction temperature of 0°C to 100°C.

[0033] The reaction requires quenching with an acid. Suitable citric acids are acetic acid or citric acid.

[0034] Suitable reaction techniques include batch reactor setups, plug-flow reactor setups, or, as a preferred technique, continuous stirred tank reactor (CSTR) setups.

[0035] The ring extension can be carried out according to Scheme 3 below. Scheme 3: [ka]

[0036] Homologization or ring extension of cyclic ketones can be carried out by using a diazo compound (e.g., diazomethane or trimethylsilyldiazomethane) in the presence of an accelerator such as a Lewis acid (e.g., BF3·Et2O, AlMe3).

[0037] Step d 1 In this case, this results in the ring extension of the bicyclic five-membered ring ketone Va to the bicyclic cyclohexyl ketone Vb.

[0038] Step d 2 In this study, the bicyclic six-membered ketone Vb can be homologized to a seven-membered bicyclic ketone under similar experimental conditions.

[0039] Homology of cyclic and acyclic ketones under different conditions has been reported in the literature (Candeias et al, Chem, Rev, 2016, 2937-2981).

[0040] a) Reduction of the ketone of formula V using a metal complex catalyst: Suitable metal complex catalysts for the reduction of the ketone of formula V are ruthenium complex catalysts or iridium complex catalysts.

[0041] These can be selected from a variety of ruthenium or iridium complex catalysts, as outlined below, including their isomers and mixtures. [ka] [ka] [ka] In the formula, each individual structure, and independently of each other, R 1 They are independent of each other, C 1-6 -alkyl, C 4-6 -Cycloalkyl, phenyl, or heteroaryl, with one or more C 1-6 -alkyl or C 1-6 - May be arbitrarily substituted with alkoxy, R 2 These are, independently of each other, hydrogen and C 1-6 -alkyl, C 4-6 -Cycloalkyl, phenyl, or heteroaryl, with one or more C 1-6 -alkyl or C 1-6 -Optionally substituted with alkoxy or two R 2 Together, they form a ring bridged by -(CH2)4- units. R 3 These are, independently of each other, hydrogen or C 1-6 -alkyl, C 4-6-Cycloalkyl, phenyl, or heteroaryl, with one or more C 1-6 -alkyl or C 1-6 - May be arbitrarily substituted with alkoxy, R 4 These are, independently of each other, hydrogen and C 1-6 -alkyl, C 4-6 -Cycloalkyl, phenyl, or heteroaryl, with one or more C 1-6 -Alkyl or C 1-6 - May be optionally substituted with an alkoxy, or both R 4 Together, they form -O-(CH2) x Forming a ring bridged by -O- units, R 5 These are, independently of each other, hydrogen and C 1-6 -alkyl, C 4-6 -Cycloalkyl, phenyl, or heteroaryl, with one or more C 1-6 -Alkyl or C 1-6 - May be optionally substituted with an alkoxy, or adjacent R 4 and R 5 Both together form -(CH)4-unit or -O-(CH2) x Forming a ring bridged by -O- units, R 6 These are, independently of each other, hydrogen or C 1-6 -It is alkyl, X is halogen, C 1-6 -alkoxy, tetrahalogenovolate, tetrakis(3,5-bis(trihalogeno-C) 1-6 It is one of the coordinated ligands or counter anions selected from -alkyl)phenyl)borate, acetylacetonate, hexahalogenophosphate, p-tolylsulfonate, methanesulfonate, or trihalogenomethanesulfonate. Y is oxygen or -CH2-, x is 1, 2, or 3. Also, the dotted ring is Q 1 is nitrogen, Q 2 This represents the aromatic ring when carbon is present. Also, the dotted ring is Q1 and Q 2 This represents a cycloalkane ring when sulfur is present.

[0042] In preferred embodiments, formulas Xh, Xk, Xm, or Xn are selected.

[0043] The preferred substituents for Xh and Xk are as follows: R 1 They are independent of each other, C 1-6 - Alkyl, phenyl, and one or more C 1-6 -alkyl or C 1-6 - May be arbitrarily substituted with alkoxy, R 2 These are, independently of each other, hydrogen or one or more C 1-6 -Phenyl optionally substituted with alkyl, or two R 2 Together, they form a ring bridged by -(CH2)4- units. R 3 These are, independently of each other, hydrogen and C 1-6 -alkyl, one or more C 1-6 - A phenyl that is optionally substituted with an alkyl group, X is either a coordinated ligand or a counteranion selected from halogens, tetrafluoroborates, tetrakis(3,5-bis(trifluoromethyl)phenyl)borates, hexafluorophosphates, or trifluoromethanesulfonates.

[0044] The preferred substituents for Xm and Xn are as follows: R 1 Each is independently a phenyl compound and one or more C 1-6 -alkyl or C 1-6 - May be arbitrarily substituted with alkoxy, R 6 These are, independently of each other, hydrogen or C 1-6 -It is alkyl, X is a coordinated ligand selected from halogens, Also, the dotted ring is Q 1is nitrogen, Q 2 This represents the aromatic ring when carbon is present.

[0045] In a more preferred embodiment, the substituents of Xh and Xk are as follows: R 1 These are methyl, phenyl, and one or more C 1-6 - May be optionally substituted with alkyl groups, R 2 These are, independently of each other, hydrogen, phenyl, or two R 2 Together, they form a ring bridged by -(CH2)4- units. R 3 These are, independently of each other, hydrogen or C 1-6 -It is alkyl, X is either a coordinated ligand or a counteranion selected from chlorides or trifluoromethanesulfonates.

[0046] Further preferred substituents for Xm and Xn are as follows: R 1 is phenyl, and optionally one or more C 1-6 - Substituted with alkyl, R 6 These are, independently of each other, hydrogen, tert-butyl, or methyl. X is a chloride, Also, the dotted ring is Q 1 is nitrogen, Q 2 This refers to the aromatic ring when carbon is present.

[0047] Suitable catalysts are typically commercially available from suppliers such as Jiuzhou Pharma, Sinocompound, or Johnson Matthey, or from catalog suppliers such as Strem or Sigma Aldrich.

[0048] The reduction of the ketone of formula V can be carried out in the presence of a reducing agent and a suitable organic solvent.

[0049] One option for the reducing agent is the use of a mixture of formic acid and a trialkylamine, preferably an alkali formate such as triethylamine or sodium formate, and the presence of a tetraalkylammonium halide, such as tetrabutylammonium bromide (TBAB).

[0050] The mixing ratio can be varied, for example, 1 to 5 equivalents of formic acid and 1 to 5 equivalents of triethylamine, or 5 equivalents of sodium formate and 0.5 equivalents of TBAB.

[0051] Typically, organic solvents selected from, for example, ethanol, toluene, acetonitrile, 2-methyltetrahydrofuran, or propylene carbonate can be used. The reaction temperature is preferably between 10°C and 100°C. The temperature can be selected between 20°C and 50°C.

[0052] As a further option, the reduction is carried out in the presence of hydrogen, with a hydrogen pressure of 1 bar to 100 bar, preferably 60 bar to 80 bar, and a reaction temperature of 10°C to 100°C, preferably 40°C to 60°C.

[0053] Suitable organic solvents are aliphatic alcohols such as ethanol.

[0054] Some catalysts require a base for activation.

[0055] Suitable bases are inorganic bases selected from alkalis or alkaline earth carbonates, bicarbonates, phosphates, hydrogen phosphates, dihydrogen phosphates, acetates, or formates, or organic bases selected from amines, alkaline alcohols, or amidines. Organic bases are generally preferred. Typical examples of organic bases are potassium tert-butyrate or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), 1,4-diazabicyclo(2.2.2)octane (DABCO), and 7-methyl-1,5,7-triazabicyclo[4.4.0]dec-5-ene (MTBD), with potassium tert-butyrate being the most preferred.

[0056] The chiral pyrrotriazole alcohol of formula I can be separated from the reaction mixture by evaporation of the solvent. Subsequent column chromatography and / or crystallization of the product yields the chiral alcohol of formula I in good yield, high purity, and high enantiomeric excess.

[0057] b) Reduction of the ketone of formula V using oxidoreductase: The appropriate oxidoreductase is the ketone of formula V. [ka] Reducing (wherein X is a halogen atom and n is an integer of 1, 2, or 3) to chiral pyrrotriazole alcohol of formula I: [ka] (In the formula, X and n are as described above, and the helical bond " [ka] "teeth," [ka] "or" [ka] " or represents a mixture of these enantiomers, The mixture is selected from those capable of forming a mixture with an enantiomer excess of at least 90%, preferably at least 95%, and more preferably at least 98%.

[0058] Asymmetric reduction is usually catalyzed by oxidoreductase in the presence of NADH or NADPH as a cofactor and is regenerated in situ. Oxidized NAD, not reduced NAD. + or NADP + It is added to the reaction system as NAD. + or NADP + The substrate-to-cofactor ratio (S / C) is typically maintained within the range of 5 to 1000, preferably between 10 and 500.

[0059] The oxidized cofactor is, in principle, continuously regenerated using a secondary alcohol as an auxiliary substrate. Typical auxiliary substrates can be selected from 2-propanol, 2-butanol, pentane-1,4-diol, 2-pentanol, 4-methyl-2-pentanol, 2-heptanol, hexane-1,5-diol, 2-heptanol, or 2-octanol, and preferably 2-propanol. Preferably, the cofactor is regenerated using the auxiliary substrate with the same enzyme that also catalyzes the target reaction. In a more preferred embodiment, the acetone formed when 2-propanol is used as an auxiliary substrate is continuously removed from the reaction mixture.

[0060] Furthermore, cofactor regeneration by additional enzymes that oxidize the native substrate and provide a reduced cofactor is also well known. Examples include secondary alcohol dehydrogenase / alcohol, glucose dehydrogenase / glucose, formate dehydrogenase / formic acid, glucose-6-phosphate dehydrogenase / glucose-6-phosphate, phosphite dehydrogenase / phosphate, and hydrogenase / molecular hydrogen. In addition, electrochemical regeneration methods are known, and chemical cofactor regeneration methods including metal catalysts and reducing agents are suitable. In particular, when glucose dehydrogenase / glucose is used for cofactor regeneration, the pH must be maintained by the addition of a controlled base to neutralize the formed gluconic acid (an oxidized byproduct of reduced nicotinamide cofactor regeneration). The substrate-to-coenzyme ratio (S / GDH) is usually maintained in the range of 5 to 1000, preferably in the range of 10 to 200.

[0061] Preferred microbial oxidoreductases are derived from yeast, bacteria, or mammalian cells.

[0062] Oxidoreductase can be applied in isolated enzyme form, whole-cell form, or sometimes in immobilized form, by one of the many conventional methods described in the literature.

[0063] In certain embodiments of the present invention, asymmetric reduction is carried out in an aqueous solvent in the presence of an organic cosolvent which can be selected from, for example, glycerol, 2-propanol, dimethyl sulfoxide, diethyl ether, tert-butyl methyl ether, diisopropyl ether, dibutyl ether, toluene, 2-methyltetrahydrofuran, ethyl acetate, butyl acetate, heptane, hexane or cyclohexene, or mixtures thereof.

[0064] The presence of an organic cosolvent is particularly advantageous because it can form a homogeneous suspension, enabling the simple separation of the desired ketone of formula V by filtration.

[0065] The reaction concentration (the concentration of the ketone of formula V and the chiral alcohol of formula I in the reaction mixture) is usually maintained in the range of 1% to 25% w / v, preferably 4% to 20% w / v.

[0066] A buffer salt is used in the reaction, which can be selected from, for example, potassium phosphate buffer, Tris-HCl buffer, bicine buffer, HEPES buffer, or PIPES buffer. The pH of the reaction is maintained in the range of 6.0 to 9.0, preferably 6.5 to 7.5.

[0067] The reaction temperature is usually maintained in the range of 10 to 50°C, preferably in the range of 20 to 30°C.

[0068] The substrate-to-enzyme ratio (S / E) is typically maintained in the range of 5 to 1000, preferably in the range of 10 to 200.

[0069] Once the reaction is complete (generally >90% conversion), the product is usually post-processed by extraction.

[0070] Depending on the ketone substrate, the preferred catalyst / cofactor / auxiliary substrate system may vary.

[0071] The following oxidoreductases have been shown to be useful in the formation of (S)-Ia (S)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol.

[0072] NADPH-dependent oxidoreductases can be selected from the KRED-NADPH-130, KRED-P1-C01, KRED-P2-D11, KRED-P2-D12, and KRED-463 types from Codexis Inc., or NADH-dependent oxidoreductases can be selected from the ADH-109, ADH-132, and ADH-172 types from c-LEcta.

[0073] The following oxidoreductases have been shown to be useful in the formation of (R)-Ia (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol.

[0074] NADPH-dependent oxidoreductase can be selected from Johnson Matthey's ADH-61 type, or NADH-dependent oxidoreductase can be selected from Codexis Inc.'s KRED-NADH-110 type.

[0075] The following oxidoreductases have been shown to be useful in the formation of (S)-Ib (S)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol.

[0076] NADPH-dependent oxidoreductase can be selected from Johnson Matthey's ADH-153 type, or NADH-dependent oxidoreductase can be selected from c-LEcta's ADH-109, ADH-110, ADH-132, and ADH-172 types.

[0077] The following oxidoreductases have been shown to be useful in the formation of (R)-Ib (R)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol.

[0078] NADPH-dependent oxidoreductases can be selected from Johnson Matthey's ADH-19, ADH-20, and ADH-61 types, or from Codexis Inc.'s KRED-425 type.

[0079] The following oxidoreductases have been shown to be useful in the formation of (S)-Ic (S)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol.

[0080] NADPH-dependent oxidoreductase can be selected from Johnson Matthey's ADH-153 type, Codexis Inc.'s KRED-P2-D11 and KRED-464 types, or NADH-dependent oxidoreductase can be selected from c-LEcta's ADH-109, ADH-110, ADH-132 and ADH-172 types.

[0081] The following oxidoreductases have been shown to be useful in the formation of (R)-Ic (R)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol.

[0082] NADPH-dependent oxidoreductases can be selected from Johnson Matthey's ADH-19, ADH-20, and ADH-61 types, or Codexis Inc.'s KRED-425 type.

[0083] Chiral pyrrolotriazole alcohol of the following formula [ka] (In the formula, X is a halogen atom, n is an integer of 1, 2, or 3, and the spiral joint " [ka] "teeth," [ka] "or" [ka] (or a mixture of these enantiomers) This is a novel compound and therefore constitutes a further embodiment of the present invention.

[0084] In a preferred embodiment, X is chlorine or bromine, preferably bromine, and n is 1, 2, or 3, and the spiral joint " [ka] "teeth," [ka] "or" [ka] This represents either a compound of these enantiomers or a mixture thereof.

[0085] In a more preferred embodiment, a) X is bromine, n is 1, and the helical bond " [ka] "teeth" [ka] This represents ".

[0086] b) X is bromine, n is 1, and the helical bond " [ka] "teeth" [ka] This represents ".

[0087] c) X is bromine, n is 2, and the helical bond " [ka] "teeth" [ka] This represents ".

[0088] d) X is bromine, n is 2, and the helical bond " [ka] "teeth" [ka] This represents ".

[0089] e) X is bromine, n is 3, and helical bond " [ka] "teeth" [ka] This represents ".

[0090] f)X is bromine, n is 3, and helical bond " [ka] "teeth" [ka] This represents ". [Examples]

[0091] [Table 1] [Table 2] TIFF2026521749000058.tif245165 TIFF2026521749000059.tif133165

[0092] Unless otherwise specified, all solvents, reagents, and compounds were purchased and used without further purification. All catalysts and ligands are typically commercially available from suppliers such as Jiuzhou Pharma, Sinocompound, Johnson Matthey, or catalog suppliers such as Strem or Sigma-Aldrich.

[0093] Analysis method: NMR method: 1 ¹H-NMR spectra were measured in CDCl3 or DMSO-d6 solution at 25°C using a Bruker AV 600 MHz, 400 MHz, or 300 MHz spectrometer equipped with a DCH cryoprobe, and the chemical shift (δ) was reported in ppm using trimethylsilyl chloride (δ=0 ppm) as an internal standard.

[0094] LC / MS method: LC / MS method for conversion from II to IVA-c and from IVA-c to Va-c, as well as for determining the purity of IVA-c and Va-c:

[0095] System: UPLC, Waters, Photodiode Array detector (PDA, Waters). Evaporative light scattering detector (ELSD, VWR 90LT). LC: Stationary phase: Agilent Zorbax EclipsePlus C18 RRHT, L=30mm, ID=2.1mm, 1.8μm, Eluent: A) Water 0.1% Formic Acid; B) MeCN 0.07% Formic Acid. Pump program: Gradient from 97A:3B to 13A:87B over 2 minutes (using UV spectrum). Run time: 2 minutes. Flow rate: 1 mL / min. Column oven temperature: 50℃. Injection volume: 2 μl. MS: Single quadrupole (Waters SQD01). m / z 150~900. Detection: PDA 210~400 nm. Residence time: II: 0.65 minutes, IVa: 0.93 minutes, IVb: 1.08 minutes, IVc: 1.15 minutes, Va: 0.70 minutes, Vb: 0.61 minutes, Vc: 0.69 minutes.

[0096] HPLC method: a) HPLC method for converting Va to Ia and determining the purity and enantiomeric excess of Ia: System: Agilent 1290. Stationary phase: Chiralpak IBN-3, L=150mm, ID=4.6mm, 3μm, Eluent: A) n-heptane, B) EtOH. Pump program: 75A:25B for 10 minutes. Runtime: 10 minutes. Flow rate: 1 mL / min. Column oven temperature: 25℃. Injection volume: 10 μl. Detection: DAD 205 nm. Residence time: Va: 13.80 min, (S)-Ia: 3.72 min, (R)-Ia: 4.36 min.

[0097] b) HPLC method for converting Vb to Ib and determining the purity and enantiomeric excess of Ib: System: Agilent 1290. Stationary phase: Chiralpak IBN-3, L=150mm, ID=4.6mm, 3μm. Eluent: A) n-heptane, B) EtOH. Pump program: 95A:5B gradient for 10 minutes, 45A:55B gradient for 5 minutes, then hold for 2 minutes, 95A:5B gradient for 0.1 minutes, then hold for 2.9 minutes. Run time: 20 minutes. Flow rate: 1 mL / min. Column oven temperature: 20℃. Injection volume: 6 μL. Detection: DAD 210 nm. Residence time: Vb: 17.0 min, (S)-Ib: 9.0 min, (R)-Ib: 9.7 min.

[0098] c) HPLC method for converting Vc to Ic and determining the purity and enantiomeric excess of Ic: System: Agilent 1290. Stationary phase: Chiralpak AD-3, L=150mm, ID=4.6mm, 3.0μm. Eluent: A) n-heptane, B) EtOH. Pump program: 90A:10B for 10 minutes. Run time: 10 minutes. Flow rate: 1 mL / min. Column oven temperature: 25℃. Injection volume: 10 μL. Detection: DAD 205 nm. Residence time: Vc (keto / enol form): 5.70 min and 6.60 min, (S)-Ic: 3.47 min, (R)-Ic: 3.81 min.

[0099] Optical rotation method: Optical rotation values ​​were obtained using an Anton Paar MCP-500 instrument. Optical rotation was measured in methanol at 20°C at the indicated concentration.

[0100] Reaction scheme Synthesis of chiral alcohols Ia-c Scheme 4: [ka]

[0101] Ring expansion reaction of Va and Vb Scheme 3: [ka]

[0102] Step A: Synthesis of ester IVA-c Example 1: a) Synthesis of methyl 3-(3,5-dibromo-1,2,4-triazol-1-yl)propionate (IVa) [ka]

[0103] 3,5-Dibromo-1H-1,2,4-triazole (40.00 g, 176.32 mmol, 1.00 equivalent) was suspended in MeTHF (90 mL). Triethylamine (5.35 g, 52.90 mmol, 0.30 equivalent) was added to the suspension. After stirring at room temperature for 5 minutes, methyl acrylate (15.94 g, 16.78 mL, 185.14 mmol, 1.05 equivalent) was added over 4 hours at 70°C, and the solution was stirred at 70°C for a further 20 hours. The solution was cooled to 20°C, washed with 27.24 g, 0.30 equivalent 2 M aqueous HCl solution, and then washed with water (20 mL). The organic phase was evaporated under vacuum to obtain the labeled compound (55.60 g, purity 92.0%, yield 92.7%) as a colorless to yellowish oil. LC / MS: 313.9, 315.9 [M+H] + ,ESI pos. 1 H-NMR (300MHz, DMSO-d6): δ 4.37 (t, J = 6.49Hz, 2H), 3.60 (s, 3H), 2.94 (t, J = 6.49Hz, 2H).

[0104] b) Synthesis of methyl 3-(3,5-dibromo-1,2,4-triazol-1-yl)propionate (IVa) on a production scale: 3,5-Dibromo-1H-1,2,4-triazole (131.0 kg, 577 mol, 1.00 equivalent) was suspended in 2-MeTHF (160 kg). Triethylamine (17.1 kg, 170 mmol, 0.30 equivalent) was added to the light suspension, and the mixture was heated to 70°C. Methyl acrylate (51.9 kg, 602 mol, 1.05 equivalent in 110 kg of 2-MeTHF) was added to the 2-MeTHF over 4 hours. The reaction mixture was stirred at 70°C for a further 24 hours. Conversion was confirmed by GC analysis (typically >98.5a%). The reaction mixture was washed with HCl (25.2 kg 25% HCl + 60 kg water) at 20°C and with water (65 kg) at 35°C. The organic phase was distilled azeotropically (200 to 100 mbar) to remove water until <200 ppm (approximately 650 kg of 2-MeTHF). The dried organic phase was diluted with 2-MeTHF and toluene (1528 kg in total) to obtain the labeled compound as a solution of approximately (ca.) 8 wt%.

[0105] Example 2: Synthesis of 4-(3,5-dibromo-1,2,4-triazol-1-yl) butyrate ethyl ester (IVb) [ka] 3,5-Dibromo-1H-1,2,4-triazole (24.70 g, 108.88 mmol, 1.00 equivalent) was dissolved in MeCN (250 mL) and DMF (25 mL). Potassium carbonate (37.62 g, 272.19 mmol, 2.50 equivalents) was added to the solution. After stirring at room temperature for 5 minutes, ethyl 4-bromobutyrate (23.36 g, 17.30 mL, 119.77 mmol, 1.10 equivalents) was added, and the suspension was stirred at 75°C for 3.5 hours, at which point LC / MS indicated that the reaction had reached complete conversion. The suspension was cooled and filtered, and the filter cake was washed several times with SiO2 (250 mL) to ensure that the product was recovered in solution. The solution was evaporated under vacuum to remove all solvent (including DMF). The crude product was purified by silica gel chromatography (eluent: dimethyl / n-heptane 5-40%) to obtain the labeled compound (32.80 g, purity 93.0%, yield 86.6%) as a colorless oil. LC / MS: 341.9, 343.9 [M+H] + ,ESI pos. 1 H-NMR (300MHz, CDCl3): δ 4.24(t,J=6.9Hz,2H),4.15(q,J=7.3Hz,2H),2.41-2.34(m,1H),2.25-2.14(m,2H),1.27(t,J=7.2Hz,3H).

[0106] Example 3: Synthesis of 5-(3,5-dibromo-1,2,4-triazol-1-yl)valerate ethyl ester (IVc) [ka] 3,5-Dibromo-1H-1,2,4-triazole (20.00 g, 88.16 mmol, 1.00 eq) was dissolved in THF (70.53 mL). Potassium carbonate (12.43 g, 89.92 mmol, 1.02 eq) was added to the solution, and the mixture was stirred at room temperature for 5 minutes. Then ethyl 5-bromovalerate (18.80 g, 13.79 mL, 89.92 mmol, 1.02 eq) was added, and the suspension was stirred at 60 °C for 20 hours. The suspension was cooled to room temperature, and the solid was filtered off through a funnel using a frit disk (G3) and washed with THF (100 mL). Evaporation of the solution under vacuum (200 mbar to 10 mbar) at 40 °C gave the title compound (33.00 g, purity 90%, yield 94.9%) as a colorless oil. LC-MS 355.9 / 357.9[M+H]+,ESI pos. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.11 - 1.24 (m, 3H) 1.45 - 1.59 (m, 2H) 1.73 - 1.87 (m, 2H) 2.27 - 2.40 (m, 2H), 3.98 - 4.11 (m, 2H) 4.11 - 4.22 (m, 2H) ppm.

[0107] Step B: Synthesis of ketones Va-c Example 4: a) Synthesis of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazol-7-one (Va)

Chemical formula

[0108] b) Synthesis of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazol-7-one (Va) on a production scale: 3-(3,5-dibromo-1,2,4-triazole-1-yl)propionate methyl ester (approximately 2-MeTHF / 8.00 wt% in toluene)-feed A (212 g / min) was added in parallel to CSTR1 (IT -60°C, Tres 10 min) together with n-hexyl lithium (2.5 M in n-heptane, 18 g / min, 1.20 equivalents). The reaction mixture was transferred from CSTR1 to CSTR2 (IT 0~ +10°C, Tres 10 min), and citric acid solution (30 wt% in water, 50 g / min, 1.5 equivalents) was added. The two-phase reaction mixture was continuously transferred from CSTR2 to batch reactor 3. The process was carried out until all feed A was consumed. The aqueous phase was separated, and the organic phase was washed with water.

[0109] (130 kg). The organic phase was concentrated in a vacuum (maximum JT 35°C) to a volume of approximately 300 L (target 20-25 wt% 2-MeTHF). The resulting suspension was cooled to -10°C and stirred at -10°C for at least 2 hours. The solid was filtered off using a frit disk (G3) and a funnel, washed with ice-cold toluene (260 kg), and dried under vacuum to obtain the marked compound (65.4 kg, 90% assay, isolation yield 50.5%) as a brown solid.

[0110] Example 5: Synthesis of 2-bromo-6,7-dihydro-5H-[1,2,4]triazole[1,5-a]pyridine-8-one (Vb) [ka] In a CSTR apparatus, ethyl 4-(3,5-dibromo-1,2,4-triazole-1-yl)butyrate (31.00 g, 90.91 mmol, 1.00 equivalent) was dissolved in MeTHF (482 mL) to obtain feed A solution (0.182 M). Feed A (6.00 mL / min) was added to CSTR1 (IT -60°C, Tres 10 min) in parallel with n-hexyl lithium (2.5 M in n-heptane, 30.11 g, 109.09 mmol, 1.20 equivalent, 0.54 mL / min) together. The reaction mixture was transferred from CSTR1 to CSTR2 (IT +10°C, Tres 10 min), and acetic acid solution (10% by weight in water, 118.18 mmol, 1.30 equivalent, 0.88 mL / min) was added. The two-phase reaction mixture was continuously transferred from CSTR2 to batch reactor 3 (total volume of the two-phase reaction mixture: 594 mL) within 80 minutes. The aqueous phase was separated, and the organic phase was washed with water (60 mL). The organic phase was concentrated to approximately 50 mL under vacuum. n-heptane (15 mL) was added, and the suspension was cooled to 0°C. The solid was filtered using a frit disk (G3) and a funnel, washed with an ice-cold mixture of MeTHF / n-heptane (1:5 v / v, 25 mL), and dried under vacuum to obtain the marked compound (4.20 g, purity 98.0%, yield 21.4%) as a pale yellow solid. LC / MS: 215.9, 217.9 [M+H] + ,ESI pos. 1 H-NMR (300MHz, DMSO-d6): δ 4.40 (t, J = 6.1, 2H), 2.77 (t, J = 6.5Hz, 2H), 2.35 (pent., J = 6.5, 6.1Hz, 2H).

[0111] Example 6: Synthesis of 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one (Vc) [ka]

[0112] Using the CSTR setup outlined, ethyl 5-(3,5-dibromo-1,2,4-triazole-1-yl)valerate (30.80 g, 86.75 mmol, 1.00 equivalent) was dissolved in MeTHF (570.2 mL) to obtain feed A solution (0.130 M). Feed A (4.00 mL / min) was added in parallel to CSTR1 (IT -60°C, Tres 10 min) along with n-hexyl lithium (2.5 M in heptane, 41.64 mL, 104.1 mmol, 1.20 equivalent, 0.25 mL / min). The reaction mixture was transferred from CSTR1 to CSTR2 (IT +5°C, Tres 10 min), and acetic acid solution (10% by weight in water, 127.82 mmol, 2.00 equivalent, 0.625 mL / min) was added. The two-phase reaction mixture was continuously transferred from CSTR2 to batch reactor 3 (within 150 minutes) (a total of 731 mL of the two-phase reaction mixture was recovered). The aqueous phase was separated, and the organic phase was washed with water (60 mL). The organic phase was concentrated under vacuum to obtain crude 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one. The product was purified by SiliaSep® HP25-40 μm silica column chromatography (Â1 / n-heptane), followed by re-purification by C18-silica gel chromatography (water / MeCN) to obtain the marked compound (315 mg, purity 95%, yield 3.1%) as a white solid. LC / MS:229.98,231.97[M+H]+,ESI pos. 1 H NMR (600MHz, CDCl3) δ ppm 4.48-4.62(m,2H),2.89-2.99(m,2H),2.16-2.28(m,2H),2.02-2.12(m,2H)ppm.

[0113] Step D: Synthesis of ketone Vb-c Example 7: Synthesis of 2-bromo-6,7-dihydro-5H-[1,2,4]triazole[1,5-a]pyridine-8-one (Vb) [ka] To a solution of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one (0.50 g, 2.48 mmol, 1.00 equivalent) dissolved in DCM (10 mL), 2M trimethylaluminum in toluene (1.49 mL, 2.97 mmol, 1.20 equivalent) was added dropwise at -78°C, followed by the addition of trimethylsilyldiazomethane in diethyl ether (1.36 mL, 2.72 mmol, 1.10 equivalent) at -78°C. The reaction mixture was stirred at -20°C for 3 hours (until LC / MS showed product formation), and a 1M HCl solution was added. The solution was extracted twice with DCM. The organic layer was dried over MgSO4 and concentrated to dryness. The crude substance was purified by flash chromatography using silica gel to obtain the labeled compound (55 mg, purity 98.0%, yield 9.6%) as a light brown solid. LC / MS: 215.96, 217.98 [M+H] + ,ESI pos. 1 H-NMR (300MHz, CDCl3): δ 4.47 (m, 2H), 2.86 (m, 2H), 2.48 (m, 2H).

[0114] Example 8: Synthesis of 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one (Vc) [ka] 2,6-Di-tert-butyl-4-methylphenol (2.24 g, 10.18 mmol, 2.20 equivalents) was dissolved in DCM (10 mL), and 2M trimethylaluminum in toluene 2M (2.78 mL, 5.55 mmol, 1.20 equivalents) was added at room temperature. The mixture was stirred for 1 hour, cooled to -78 °C, and then 2-bromo-6,7-dihydro-5H-[1,2,4]triazolo[1,5-a]pyridin-8-one (1.00 g, 4.63 mmol, 1.00 equivalent) was dissolved in dichloromethane (10 mL), and then a 2M solution of TMS-diazomethane 2M in diethyl ether (2.55 mL, 5.09 mmol, 1.10 equivalents) was added. The reaction was stirred at -78 °C for 2 hours until LC / MS indicated the formation of the product, and then 1M HCl solution was added. The reaction was extracted twice with DCM, the organic layer was dried over MgSO4, and concentrated to dryness under vacuum. The crude material was purified by flash chromatography on silica gel (40 g, 0 - 20% EtOAc in n-heptane). The resulting compound was further purified again by flash chromatography on C18 (50 g, 10 - 95% water in MeCN) to obtain the title compound (106 mg, purity 95.0%, yield 10.0%) as a light brown solid. LC / MS: 229.98, 231.97 [M+H] + , ESI pos. 1 1H-NMR (300 MHz, CDCl3): δ 4.54 (m, 2H), 2.93 (m, 2H), 2.23 (m, 2H), 2.09 (m, 2H).

[0115] Step C: Synthesis of alcohols Ia-c by metal-catalyzed reduction Example 9.1: (S)-2-Bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazol-7-ol ((S)-Ia) synthesis

Chemical formula

[0116] Example 9.2: Synthesis of (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol ((R)-Ia) [ka] In a glove box (<1 ppm O2), 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one (100 mg, 495.03 μmol, 1.0 equivalent) was weighed into a 10 mL Schlenk flask and suspended in MeCN (0.5 mL). Then, formic acid (57 mg, 47.5 μL, 1.24 mmol, 2.5 equivalents) and triethylamine (62.6 mg, 86.1 μL, 618.8 μmol, 1.25 equivalents) were added, followed by Ru-466 (3.15 mg, 4.95 μmol, 0.01 equivalent) and acetonitrile (0.5 mL). The Schlenk flask was sealed with a septum and removed from the glove box, and the reaction mixture was stirred at Tj 32 °C for 19 hours (LC / MS showed complete conversion). The yellow reaction mixture was cooled to room temperature, the solvent was removed under vacuum, and the mixture was dried to obtain the crude product. The crude product was purified by column chromatography using Â1 / n-heptane to obtain 92 mg of the labeled compound as a white solid (purity >99%, yield 90.1%, (S):(R)-Ia=0.5:99.5). [α] D 20 :-11.0°(MeOH, c=1.053).

[0117] Examples 9.3-9.11 Synthesis of (S)- or (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol(Ia) [ka] Va (5 mg, 25 μmol) was reduced to Ia under the conditions listed in Table E9, similar to Example 9.1, which uses the HTE setting (48-96 well plate format). [Table 3]

[0118] Example 10.1: Synthesis of (S)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol ((S)-Ic) [ka] In a glove box (<1 ppm O2), a Schlenk tube was filled with 2-bromo-6,7-dihydro-5H-[1,2,4]triazolo[1,5-a]pyridine-8-one (100 mg, 462.9 μmol, 1.0 equivalent), toluene (2 mL), formic acid (106.5 mg, 88.8 μL, 2.1 mmol, 5.0 equivalent), and trimethylamine (234.2 mg, 322.6 μL, 2.3 mmol, 5.0 equivalent), and stirred for 2 minutes to obtain a solution. Ru-461 (14.72 mg, 23.14 μmol, 0.05 equivalent) was added to this solution, the Schlenk tube was sealed with a septum, and removed from the glove box. The flask was connected to an argon supply tube, and the yellow reaction mixture was stirred in an oil bath at Tj 42°C for 4 hours to obtain complete conversion (LC / MS analysis, (S):(R)-Ib:98.6:1.4). The yellow reaction mixture was cooled to room temperature, the solvent was removed under vacuum, and the mixture was dried to obtain the crude product (114 mg). The crude product was purified by column chromatography using toluene / n-heptane to obtain the marked compound (72 mg, purity 95.0%, yield 71.0%, (S):(R)-Ib=98.7:1.3) as a white solid. 1H-NMR(600MHz,DMSO-d6):δ 5.85(br s,1H),4.70(t,J=5.1,1H),4.14(s,1H),4.00(ddd,J=12.9,7.9,5.2Hz,1H), 2.08-2.17(m,1H),1.96-2.03(m,1H),1.88-1.94(m,1H),1.80-1.85(m,1H). [α] D 20 : +0.525° (MeOH, c=1.047).

[0119] Example 10.2: Synthesis of (R)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol ((R)-Ib [ka] In a glove box (<1 ppm O2), an autoclave was loaded with 2-bromo-6,7-dihydro-5H-[1,2,4]triazole[1,5-a]pyridine-8-one (100 mg, 462.9 μmol, 1.0 equivalent), KOtBu (1.00 mg, 9.30 μmol, 0.02 equivalent), Ir-15 (6.74 mg, 9.3 μmol, 0.02 equivalent), and ethanol (1.5 mL) to obtain an orange suspension. The autoclave was sealed, pressurized with argon at 7 bar, and removed from the glove box. The autoclave was connected to a hydrogenation line, and the line and autoclave were flushed with hydrogen. The autoclave was then pressurized with hydrogen at 70 bar and stirred at Tj 52°C for 21 hours. The autoclave was then returned to ambient temperature and the pressure was released. The autoclave was opened and a sample was taken for analysis ((S):(R)-Ib=5:95). The reaction mixture was transferred to a round-bottom flask, and the solvent was removed under vacuum to obtain 108 mg of the crude labeled compound as a reddish oil. The crude product was purified by column chromatography using dimethyl / n-heptane to obtain the labeled compound (71 mg, purity 99.0%, yield 77.0%, (S):(R)-Ib=5.3:94.7) as a white solid. MS(EI + ): m / z 217.0 [MH] + . [α] D 20 :-3.836°(MeOH, c=1.040).

[0120] Examples 10.3-10.10: Synthesis of (S)- or (R)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol ((S)- / (R)-Ib) [ka] Vb (2 mg, 8.7 μmol) was reduced to Ib under the conditions listed in Table E10, similar to Example 10.1 using the HTE setting (48-96 well plate format). [Table 4]

[0121] Example 11.1: Synthesis of (S)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol ((S)-Ic) [ka] In a glove box (<1 ppm O2), 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one (50 mg, 217.3 μmol, 1.00 equivalent) and Ru-461 (1.38 mg, 2.17 μmol, 0.01 equivalent) were weighed into 10 mL Schlenk tubes and dissolved in MeCN (1 mL). Then, a pre-formed mixture of MeCN (0.5 mL), formic acid (25 mg, 20.8 μL, 543.3 μmol, 2.5 equivalents) and triethylamine (27.50 mg, 37.8 μL, 271.67 μmol, 1.25 equivalents) was added to the solution. The flask was sealed with a septum and removed from the glove box. The yellow reaction mixture was stirred in an oil bath at Tj 33°C for 22 hours to obtain complete conversion (LC / MS analysis). The yellow reaction mixture was cooled to room temperature, the solvent was removed under vacuum, and the mixture was dried to obtain the crude product. The crude product was purified by column chromatography using toluene / n-heptane to obtain the marked compound (45 mg, purity >99%, yield 89.2%, (S):(R)-Ic=93.6:6.4) as a white solid. MS(EI + ):m / z 231(M + ). 1 H-NMR(600MHz, CDCl3):δ 4.93(dd,J=8.8,2.6Hz,1H),4.34-4.46(m,1H),4.07-4.18(m,1H),2.67 -3.68(m,1H),2.13-2.28(m,1H),1.95-2.04(m,1H),1.72-1.94(m,4H). [α] D 20 :+11.296°(MeOH, c=0.108)

[0122] Example 11.2: Synthesis of (R)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol ((R)-Ic) [ka] In a glove box (<1 ppm O2), 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one (60 mg, 260.8 μmol, 1.0 equivalent) and Ru-466 (1.66 mg, 2.61 μmol, 0.01 equivalent) were weighed into 10 mL Schlenk tubes and dissolved in MeCN (1.2 mL). Then, a pre-formed mixture of MeCN (0.6 mL), formic acid (30 mg, 25 μL, 652 μmol, 2.5 equivalents), and triethylamine (33 mg, 45.4 μL, 326 μmol, 1.25 equivalents) was added to the solution. The flask was sealed with a septum and removed from the glove box. The yellow reaction mixture was stirred in an oil bath at Tj 32°C for 19 hours to obtain complete conversion (LC / MS analysis). The yellow reaction mixture was cooled to room temperature, the solvent was removed under vacuum, and the mixture was dried to obtain the crude product. The crude product was purified by column chromatography using toluene / n-heptane to obtain the labeled compound (56 mg, purity >99%, yield 92.5%, (S):(R)-Ic=6.2:93.8) as a white solid. [α] D 20 :-1.669°(MeOH, c=1.067).

[0123] Examples 11.3-11.10: Synthesis of (S)- and (R)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol ((S)- / (R)-Ic) [ka] Similar to Example 11.1 using the HTE setting (48-96 well plate format), Vc (2 mg, 8.7 μmol) was reduced to Ic under the conditions listed in Table E11. [Table 5]

[0124] Step C: Synthesis of alcohols Ia-c by enzyme-catalyzed reduction Examples 12.1-12.10: Synthesis of (S)- and (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol ((S)- / (R)-Ia) [ka] A series of oxidoreductases were screened to identify oxidoreductases that can reduce 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one to (S)- or (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol.

[0125] The reaction mixture was prepared in 100 mM potassium phosphate buffer at pH 7. NADP + (1g / L), NAD + (1 g / L), oxidoreductase (2 g / L), D-glucose (100 mM), and glucose dehydrogenase GDH-105 (Codexis) (0.1 g / L), as outlined in Table E12, were added to the reaction mixture from stock prepared in water. The reaction substrates were dissolved in dimethyl sulfoxide at a concentration of 100 g / L and dispensed into the reaction mixture at a final concentration of 5 g / L. The final volume of the reaction mixture was 0.5 mL.

[0126] The reaction mixture was incubated at room temperature for 18 hours. The reaction was quenched by adding 1 volume of acetonitrile. The reaction mixture was then centrifuged at 3220°C relative centrifugal force (RCF) for 5 minutes, and the clarified solution was transferred to a new glass vial for analysis.

[0127] The best enzyme selection for the conversion is reported in Table E12. [Table 6]

[0128] Example 12.11: Synthesis of (S)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol ((S)-Ia) [ka] Similar to Example 12.6, Codexis oxidoreductase KRED-P2-D11 (3 mg), D-(+)-glucose monohydrate (9 mmol), commercially available glucose dehydrogenase GDH-105 (15 mg) from Codexis, and oxidative cofactor NADP from Roche Diagnostics were used. + A reaction solution was prepared in 25.5 mL of aqueous buffer (100 mM potassium phosphate buffer, pH 6.5) containing (15 mg) with gentle stirring. The reaction solution was incubated at ambient temperature (23°C) and stirred for 5 minutes. Then, reduction was initiated by adding 1.5 g (7.43 mmol) of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one (Va) in 3 mL of toluene. The pH was adjusted and maintained constant during the reaction by adding 1 M NaOH via Metrohm pH Stat (Metrohm 902 Titrando).

[0129] At ambient temperature, complete conversion (IPC: >99.9 area % of product) was achieved within 8 hours at a constant pH, consuming 7.43 mL of NaOH. Toluene was removed from the reaction by evaporation under reduced pressure. The reaction was then filtered through filter paper. After filtration, sodium carbonate (20 g) and 2-methyltetrahydrofuran (100 mL) were added to the reaction and mixed vigorously, causing spontaneous phase separation. The separated aqueous phase was extracted again with 2-methyltetrahydrofuran (100 mL), and the combined phase was dried over MgSO4, filtered, and evaporated under vacuum at 40°C to obtain the marked compound (1.15 g, purity >95%, yield 76.0%, (S):(R)-Ia>99.9:0.1%, (R)-Ia and Va not detected) as a grayish-white solid. LC / MS: 203.98 (M+H) + .ESI pos. 1H-NMR(600MHz,DMSO-d6)δ 5.99(br s,1H),5.05(dd,J=6.9,3.2Hz,1H),4.23(dddd,J=10.8,8.5,4.9,0.9Hz,1H),4.04(ddd,J=10.9,8 .6,5.0Hz,1H),2.93(dddd,J=13.4,8.5,7.8,5.0Hz,1H),2.32(dddd,J=13.4,8.6,4.9,3.8Hz,1H).

[0130] Example 12.12: Synthesis of (R)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol ((R)-Ia) [ka] Similar to Example 12.12, Codexis oxidoreductase KRED-NADH-110 (250 mg), D-(+)-glucose monohydrate (27.2 mmol), commercially available glucose dehydrogenase GDH-105 (250 mg) from Codexis, and oxidative cofactor NADP from Roche Diagnostics were used. +A reaction solution was prepared in 32.1 mL of aqueous buffer (100 mM potassium phosphate buffer, pH 6.5) containing (250 mg) with gentle stirring. The reaction solution was incubated at ambient temperature (23°C) and stirred for 5 minutes. Then, reduction was initiated by adding 5 g (24.75 mmol) of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one (Va) in 7.5 mL of toluene. The pH was adjusted and maintained constant during the reaction by adding 1 M NaOH via Metrohm pH Stat (Metrohm 902 Titrando). Complete conversion (IPC: >99.9 area % of product) was achieved at a constant pH within 8 hours at ambient temperature, consuming 12.38 mL of NaOH. Toluene was removed from the reactants by evaporation under reduced pressure. The reactants were then filtered through filter paper. After filtration, sodium carbonate (20 g) and 2-methyltetrahydrofuran (100 mL) were added to the reaction mixture and mixed vigorously, causing the phases to spontaneously separate. The separated aqueous phase was extracted again with 2-methyltetrahydrofuran (100 mL), and the combined phases were dried over MgSO4, filtered, and evaporated under vacuum at 40°C to obtain the marked compound (1.64 g, purity >95%, yield 32.0%, (S):(R)-Ia=1.4:98.6, Va not detected) as a grayish-white solid. LC / MS: 203.98 (M+H) + .ESI pos. 1H NMR(600MHz,DMSO-d6)δ ppm 5.99(br s,1H),5.05(br dd,J=6.9,3.2Hz,1H),4.23(dddd,J=10.8,8.5,4.9,0.9Hz,1H),4.04(ddd,J=10.8,8.6,5.0 Hz,1H),2.93(dddd,J=13.4,8.5,7.8,5.0Hz,1H),2.32(dddd,J=13.4,8.6,4.9,3.8Hz,1H).

[0131] Example 12.13: Large-scale synthesis of (S)-2-bromo-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-7-ol ((S)-Ia) Similar to Example 12.11, 11.75 kg of KH2PO4 and 11.76 kg of K2HPO4, and 385 g of NADP were added in a reactor under an inert atmosphere. + The reaction product was prepared by mixing 1.54 kg of disodium salt and KRED-P2-D11 (Codexis) in 1,100 L of water. Then, 240 L of isopropanol was added to the reactor. The pH of the solution was 7.0, and the solution was incubated at 22°C. The reaction was initiated by adding 77 kg (442 mol) of 2-bromo-5,6-dihydropyrrolo[1,2-b][1,2,4]triazole-7-one (Va) in eight divided doses of 9.8 kg to 9.6 kg at 1-hour intervals. The reaction was completed after 12 hours (IPC: >99.9% conversion rate, 97.9% Ia, (S): (R)-Ia >99.9: 0.1%).

[0132] Isopropanol and acetone were removed under reduced pressure at 55°C until 560 kg of distillate was removed. 1150 kg of water was added, and the reaction mixture was stirred for 30 minutes. The pH was then set to 2.0 by adding 53 kg of 20% sulfuric acid in water, and the mixture was then stirred at 55°C for 45 minutes to precipitate the enzyme.

[0133] The solution was filtered through a 3M® Zeta Plus® filter cartridge, and then the feeder was rinsed twice with a mixture of 231 kg of water and 11.5 kg of 20% sulfuric acid in water, and this solution was also passed through the filter.

[0134] The filtered solution was heated to 100°C, and the water was distilled until approximately 720 kg of the solution remained in the autoclave. The temperature was lowered to 25°C, and 1,150 kg of 2-methyltetrahydrofuran was added to the reactor to extract compound Ia into the organic phase. The extraction procedure was repeated twice by removing the water fraction and adding 493 kg of 2-methyltetrahydrofuran. Subsequently, the solution of Ia in 2-methyltetrahydrofuran was filtered through a zetacarbon filter.

[0135] The 2-methyltetrahydrofuran solution was heated to 110°C, and the solvent was removed under vacuum until 380 kg of the solution remained in the reactor. This solution was cooled to 80°C, and then further cooled to 20°C at a rate of 20°C / hour. Next, 237 kg of n-heptane was added to the reactor over 45 minutes, and the solution was stirred for 2 hours. The suspension was then cooled to 0°C at a rate of 10°C / hour and stirred for 2 hours.

[0136] The suspension was centrifuged, and the crystals were washed with 264 kg of n-heptane. Finally, the crystals were dried in an oven at 50°C under total vacuum.

[0137] Compound Ia was isolated as off-white crystals in a reaction yield of 84% (IPC purity 99.7%, (S):(R)-Ia>99.9:0.1%).

[0138] Examples 13.1-13.9: Synthesis of (S)- and (R)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridine-8-ol ((S)- / (R)-Ib) [ka] A series of oxidoreductases were screened to identify oxidoreductases capable of reducing 2-bromo-6,7-dihydro-5H-[1,2,4]triazole[1,5-a]pyridin-8-one to (8S) or (8R)-2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]pyridin-8-ol. The reaction products were prepared in 100 mM potassium phosphate buffer at pH 6.5. NADP + (1g / L), NAD +(1 g / L), oxidoreductase (0.08 g / L), D-glucose (100 mM), and glucose dehydrogenase GDH-105 (Codexis) (0.02 g / L) were added to the reaction mixture from stock prepared in water. The reaction substrates were dissolved in dimethyl sulfoxide at a concentration of 40 g / L and dispensed into the reaction mixture at a final concentration of 2 g / L. The final volume of the reaction mixture was 0.5 mL. The reaction mixture was incubated at room temperature for 2–16 hours. The reaction was quenched by adding 1 volume of MeCN. The reaction mixture was then centrifuged at 3220 rcf for 5 minutes, and the clarified solution was transferred to a new glass vial for analysis. The enzymes selected for conversion are reported in Table E13. [Table 7]

[0139] Examples 14.1-14.11: Synthesis of (S)- and (R)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol ((S)- / (R)-Ic) [ka] To identify oxidoreductases capable of reducing 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazole[1,5-a]azepine-9-one to (9S) or (9R)-2-bromo-6,7,8,9-tetrahydro-5H-[1,2,4]triazole[1,5-a]azepine-9-ol, a panel of oxidoreductases was screened. Reactants were prepared in 100 mM potassium phosphate buffer at pH 6.5. NADP + (1g / L), NAD +(1 g / L), oxidoreductase (0.08 g / L), D-glucose (100 mM), and glucose dehydrogenase GDH-105 (Codexis) (0.02 g / L) were added to the reaction mixture from stock prepared in water. The reaction substrates were dissolved in DMSO at a concentration of 40 g / L and dispensed into the reaction mixture at a final concentration of 2 g / L. The final volume of the reaction mixture was 0.5 mL. The reaction mixture was incubated at room temperature for 2–16 hours. The reaction was quenched by adding 1 volume of MeCN. The reaction mixture was then centrifuged at 3220 rcf for 5 minutes, and the clarified solution was transferred to a new glass vial for analysis. The enzymes selected for conversion are reported in Table E14. [Table 8]

Claims

1. Equation I 【Chemistry 1】 (In the formula, X is a halogen atom, n is an integer of 1, 2, or 3, and the helical bond " 【Chemistry 2】 "teeth," 【Transformation 3】 "or" 【Chemistry 4】 (or a mixture of these enantiomers) A process for the preparation of chiral pyrrotriazole alcohols of formula V 【Transformation 5】 (In the formula, X and n are as described above.) The ketones, a) Metal complex catalysts in the presence of a reducing agent, or b) Oxidoreductase A process comprising reduction to form a chiral alcohol of formula I.

2. The process according to claim 1, wherein the metal complex catalyst is a ruthenium or iridium complex catalyst.

3. The metal complex catalyst is defined as follows: 【Transformation 6】 【Transformation 7】 【Transformation 8】 (In the formula, each individual structure, and independently of each other, R １ They are independent of each other, C １－６ - Alkyl, C ４－６ - Cycloalkyl, phenyl, or heteroaryl, with one or more C １－６ - Alkyl or C １－６ - May be arbitrarily substituted with alkoxy, R ２ is, independently of one another, hydrogen, C １－６ -alkyl, C ４－６ -cycloalkyl, phenyl or heteroaryl, optionally substituted with one or more C １－６ -alkyl or C １－６ -alkoxy, or two Rs ２ together form a ring bridged by a -(CH ２ ) ４ -unit, R ３ These are, independently of each other, hydrogen or C １－６ - Alkyl, C ４－６ - Cycloalkyl, phenyl, or heteroaryl, with one or more C １－６ - Alkyl or C １－６ - May be arbitrarily substituted with alkoxy, R ４ These are, independently of each other, hydrogen and C １－６ - Alkyl, C ４－６ - Cycloalkyl, phenyl, or heteroaryl, with one or more C １－６ - Alkyl or C １－６ - May be optionally substituted with an alkoxy, or both R ４ Together, -O-(CH ２ ) ｘ Forming a ring bridged by -O- units, R ５ These are, independently of each other, hydrogen and C １－６ - Alkyl, C ４－６ - Cycloalkyl, phenyl, or heteroaryl, with one or more C １－６ - Alkyl or C １－６ - May be optionally substituted with an alkoxy, or adjacent R ４ and R ５ Both together, -(CH) ４ - Unit or -O- (CH ２ ) ｘ Forming a ring bridged by -O- units, R ６ These are, independently of each other, hydrogen or C １－６ -It is alkyl, X is halogen, C １－６ - Alkoxy, tetrahalogenovolate, tetrakis(3,5-bis(trihalogeno-C) １－６ -A coordinated ligand or counter anion selected from alkyl)phenyl)borate, acetylacetonate, hexahalogenophosphate, p-tolylsulfonate, methanesulfonate, or trihalogenomethanesulfonate. Y is oxygen or -CH ２ - and x is 1, 2, or 3. The dotted ring is Q １ is nitrogen, Q ２ When it is carbon, it represents an aromatic ring. The dotted ring is Q １ and Q ２ (When sulfur is present, it represents a cycloalkane ring.) The process according to claim 1 or 2, wherein the catalyst is selected from a ruthenium complex catalyst or an iridium complex catalyst.

4. The process according to any one of claims 1 to 3, wherein the metal complex catalyst is selected from metal catalyst complexes of formula Xh, Xk, Xm, or Xn.

5. The process according to any one of claims 1 to 4, wherein the reducing agent is a mixture of formic acid and a trialkylamine, or hydrogen.

6. The process according to claim 5, wherein the reaction of formic acid with a mixture of trialkylamine is carried out in the presence of an organic solvent at a reaction temperature of 10°C to 100°C.

7. The process according to claim 5, wherein the reaction with hydrogen is carried out in an organic solvent, at a hydrogen pressure of 1 bar to 100 bar and a reaction temperature of 10°C to 100°C.

8. The process according to claim 1, wherein the selected oxidoreductase has the ability to reduce the ketone of formula V to form a chiral pyrrotriazole alcohol of formula I with an enantiomer excess of at least 90%, preferably at least 95%, and more preferably at least 98%.

9. The process according to claim 1 or 8, wherein the enzymatic reduction is carried out in the presence of NADH or NADPH as a cofactor.

10. The process according to any one of claims 1, 8, or 9, wherein a cofactor is regenerated with an auxiliary substrate.

11. The process according to claim 10, wherein the auxiliary substrate is a secondary alcohol, preferably 2-propanol.

12. The process according to any one of claims 1, 8 to 11, wherein the enzymatic reduction is carried out in an aqueous medium at a temperature of 10°C to 50°C in the presence of an organic cosolvent.

13. The ketone in formula V is, a) Equation II 【Chemistry 9】 (In the formula, X is a halogen atom.) The 3,5-dihalogen-1H-1,2,4-triazole, a １ Formula IIIa 【Chemistry 10】 (In the formula, R is C) １－４ (It is alkyl.) By Michael addition using acrylic ester, or a ２ Formula IIIb 【Chemistry 11】 (In the formula, R is C) １－４ (It is an alkyl group, n is an integer of 1, 2, or 3, and X is a halogen.) Alkylation by alkylation with halogenated carboxylic acid alkyl esters, Formula IV 【Chemistry 12】 (In the formula, R, n, and X are as described above.) Conversion of to 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester, and b) Ring closure of the 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester of formula IV using organometallic reagents, resulting in formula V 【Chemistry 13】 (In the formula, X and n are as described above.) The process according to claim 1, which can be prepared by the formation of a ketone.

14. Equation I 【Chemistry 14】 (In the formula, X is a halogen atom, n is an integer of 1, 2, or 3, and the helical bond " 【Chemistry 15】 "teeth," 【Chemistry 16】 "or" 【Chemistry 17】 (or a mixture of these enantiomers) Chiral pyrrolotriazole alcohol.

15. X is chlorine or bromine, preferably bromine, n is 1, 2 or 3, and a helical bond " [Chemistry 18] "but" 【Chemistry 19】 "or" 【Chemistry 20】 The chiral pyrrotriazole alcohol according to claim 14, which represents "" or a mixture of these enantiomers.

16. a) X is bromine, n is 1, and the helical bond " 【Chemistry 21】 "teeth" 【Chemistry 22】 This represents ", b) X is bromine, n is 1, and the helical bond " 【Chemistry 23】 "teeth" 【Chemistry 24】 This represents ", c) X is bromine, n is 2, and the helical bond " 【Chemistry 25】 "teeth" 【Chemistry 26】 This represents ", d) X is bromine, n is 2, and the helical bond " 【Chemistry 27】 "teeth" 【Chemistry 28】 This represents ", e) X is bromine, n is 3, and the helical bond " 【Chemistry 29】 "teeth" 【Transformation 30】 This represents ", f) X is bromine, n is 3, and the helical bond " 【Chemistry 31】 "teeth" 【Chemistry 32】 Representing " The chiral pyrrotriazole alcohol according to claim 14 or 15.

17. Formula IV 【Transformation 33】 (In the formula, R is C) １－４ (It is an alkyl group, n is an integer of 1, 2, or 3, and X is a halogen.) 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester.

18. The 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester according to claim 17, wherein X is bromine.

19. The 3,5-dihalogen 1,2,4-triazole-carboxylic acid ester according to claim 17 or 18, wherein R is methyl or ethyl.

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