Ethyl (6E)-7-[2-cyclopropyl-4-(4-fluorophenyl)-3-quinolinyl]-5-hydroxy-3-oxo-6-heptenoate

Rhodotorula aurantiaca IFO0754 and Rhodotorula glutinis var dairenensis IFO0415 were respectively inoculated in 2.5 mL of a liquid culture medium having the same composition as that of Example 1. At 27° C. under aerobic conditions, Rhodotorula aurantiaca was incubated for 24 hours and Rhodotorula glutinis var dairenensis was incubated for 48 hours, respectively. The obtained culture solution was centrifuged in an amount of 1 ml at a time to collect microbial cells. Then, 0.2 ml of a 100 mM potassium phosphate buffer (pH 7.0) was added in the microbial cells to suspend them completely, followed by adding 20 μl of a 50percent(w/v) glucose solution, 20 μl of a mixed solution of 2 g/L NADP and NAD, and 30 μl of a 10 g/L DOXE (DMSO solution) in the suspension and then stirring the mixture well to initiate a reaction for 12 hours at 27° C. under the aerobic conditions. [0162] After terminating the reaction, the extraction with ethyl acetate and the TLC were performed in the same way as those of Example 1. An Rf spot portion corresponding to the 5-MOLE or the 3-MOLE and an Rf spot portion corresponding to the DOLE were scraped off, respectively. [0163] Thereafter, DOLE and MOLE were analyzed using a high-performance liquid chromatography (HPLC) under the following conditions. [0164] The following is the conditions of DOLE. [0165] Column: CHIRALCEL AD (manufactured by Daicel Chemical Industries, Ltd.) [0166] Eluting solution: Hexane/ethanol=95/5 [0167] Flow rate: 1 ml/min. [0168] Detection: UV 254 nm [0169] Temperature: Room temperature [0170] Further, the following is the conditions of MOLE. [0171] Column: MCIGEL CHP2MGM (4.6.x.150 mm) (manufactured by Mitsubishi Chemical Corporation) [0172] Eluting solution: Methanol/acetonitrile/water/phosphoric acid=800/100/100/0.5 [0173] Flow rate: 0.6 ml/min. [0174] Detection: UV 254 nm [0175] Temperature: 60° C. [0176] The results are listed in Table 3.60% NaH 80·35 g (2.01 mol, 2.55 eq) and 2.5L anhydrous THF (Tetrahydrofuran) was added to in a 10 L three-necked flask, and the mixture was stirred under a nitrogen atmosphere at a temperature of -25 C, and 174.28 g of the compound II (1.34 mil, 1.7 eq), was added dropwise and stirred under nitrogen for 2 h, A 2.5 M solution of n-butyllithium in n-hexane was added dropwise 662 mL (1.65 mol, 2 leq), stirred for 2 h, Finally, the compound I 250g (787.75 mmol, leq) was added dropwise 2.5 L of anhydrous THF solution, stirred for 3 h, TLC detection of the basic reaction is complete. The reaction was quenched by the addition of 2 L of saturated aqueous ammonium chloride solution, Ether extraction 1L X 3 times, combined with organic phase, Purified washed 1L X 2 times, Na2SO4 dry, filtered, In 30 C ~ 40 C water bath rotation concentrated to dry, The crude product of compound m was 325 g. Yield 92.2%, determination of Chemical Purity by 87.2%.320 g (715.08 mmol, 1 eq) of crude compound of II and 2.2 L of anhydrous THF were added And 1.1 L anhydrous MuOEta Of the mixed solvent was added to a 5 L three-necked flask, stirred and dissolved under a nitrogen atmosphere at -55 C, A solution of 4M solution of diethylmethoxyborane in tetrahydrofuran was added dropwise to a solution of 304 mL (1.22 momicronl, 1.7 eq) stirring for 3h, and then adding 96% NaBH4 73.27g (1.86mol, 2.6eq), stirring 5h, TLC detection of basic reaction completely. After adding 1L H2O, 1L X 3 times was extracted with methyl tert-butyl ether, the combined organic phase, 1L saturated aqueous solution of sodium bicarbonate were washed once, 1L of purified water was washed once again, and the organic phase was washed at 30 C ~ 40 C water bath spin to dry. The residue was dissolved in 500 mL of MuOEta and then concentrated to dryness at 30 C to 40 C. The reaction was repeated 12 times to give 300.5 g of crude product of compound IV. Yield 93.5%, HPLC detection Chemical purity 92%.After flushing a reactor flask with nitrogen, In a 500-ml four-neck flask equipped with a stirrer, a dropping funnel, and a thermo-meter, 5.02 g (11.22 mmol) of Rhodotorula aurantiaca IFO0754 and Rhodotorula glutinis var dairenensis IFO0415 were respectively inoculated in 2.5 mL of a liquid culture medium having the same composition as that of Example 1. At 27 C. under aerobic conditions, Rhodotorula aurantiaca was incubated for 24 hours and Rhodotorula glutinis var dairenensis was incubated for 48 hours, respectively. The obtained culture solution was centrifuged in an amount of 1 ml at a time to collect microbial cells. Then, 0.2 ml of a 100 mM potassium phosphate buffer (pH 7.0) was added in the microbial cells to suspend them completely, followed by adding 20 mul of a 50%(w/v) glucose solution, 20 mul of a mixed solution of 2 g/L NADP and NAD, and 30 mul of a 10 g/L DOXE (DMSO solution) in the suspension and then stirring the mixture well to initiate a reaction for 12 hours at 27 C. under the aerobic conditions. [0162] After terminating the reaction, the extraction with ethyl acetate and the TLC were performed in the same way as those of Example 1. An Rf spot portion corresponding to the 5-MOLE or the 3-MOLE and an Rf spot portion corresponding to the DOLE were scraped off, respectively. [0163] Thereafter, DOLE and MOLE were analyzed using a high-performance liquid chromatography (HPLC) under the following conditions. [0164] The following is the conditions of DOLE. [0165] Column: CHIRALCEL AD (manufactured by Daicel Chemical Industries, Ltd.) [0166] Eluting solution: Hexane/ethanol=95/5 [0167] Flow rate: 1 ml/min. [0168] Detection: UV 254 nm [0169] Temperature: Room temperature [0170] Further, the following is the conditions of MOLE. [0171] Column: MCIGEL CHP2MGM (4.6×150 mm) (manufactured by Mitsubishi Chemical Corporation) [0172] Eluting solution: Methanol/acetonitrile/water/phosphoric acid=800/100/100/0.5 [0173] Flow rate: 0.6 ml/min. [0174] Detection: UV 254 nm [0175] Temperature: 60 C. [0176] The results are listed in Table 3.Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12Each kind of strains shown in Table 12 was incubated in the same way as that of Example 1, except that 5-MOLE was used instead of DOXE. A spot (developing solvent; hexane:ethyl acetate=1:1, Rf=0) corresponding to the compound (IV) (which is a compound, in the formula, R=hydrogen: hereinafter, abbreviated as DCOOH) on the TLC was scraped off and was then eluted with 0.25 mL of isopropanol. After centrifugation, a supernatant was subjected to a high-performance liquid chromatography (HPLC) under the same conditions as those of Example 15, to analyze the optical purity. The results are listed in Table 12

Molecular Weight:447.5
XLogP3:4.3
Hydrogen Bond Donor Count:1
Hydrogen Bond Acceptor Count:6
Rotatable Bond Count:10
Exact Mass:447.18458647
Monoisotopic Mass:447.18458647
Topological Polar Surface Area:76.5
Heavy Atom Count:33
Complexity:700
Undefined Atom Stereocenter Count:1
Defined Bond Stereocenter Count:1
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes