Method for synthesizing the small molecule compound BRD0539

Abstract

To provide a method for readily synthesizing BRD0539, which is an inhibitor of CRISPR-Cas9.SOLUTION: For example, a following synthesizing method is illustrated.SELECTED DRAWING: None

Description

[Technical field]

[0001] This invention relates to a novel method for synthesizing the small molecule compound BRD0539, an inhibitor of CRISPR-Cas9. [Background technology]

[0002] CRISPR-Cas9 was discovered in bacteria and archaea as an adaptive immune system of microorganisms that confers acquired immunity to foreign viruses and plasmids, and is an immune mechanism in which when a virus invades a bacterium, the bacterium incorporates the viral gene into its own genes and memorizes it, and if the same virus invades again, it cuts and eliminates the gene (Non-Patent Document 1, etc.). By applying this, it is possible to recognize any base sequence in DNA and delete or replace that part, and it has attracted attention as a genome editing technology that is more accurate and simpler than conventional techniques (Non-Patent Documents 2, 3, etc.). Meanwhile, in 2019, a small molecule compound BRD0539 (Chemical Formula 1) that acts as an inhibitor of CRISPR-Cas9 was discovered (Non-Patent Document 4). By using this inhibitor, BRD0539, it becomes possible to more precisely control the activity of CRISPR-Cas9. [ka]

[0003] The synthetic route for BRD0539 has already been reported (Non-Patent Document 5, Patent Document 1, etc.). The outline of the first reported synthesis method of BRD0539 (Non-Patent Document 5) is shown in the following formula (Chemical Formula 4). That is, 4-bromoaniline is used as a starting material, and tricyclic compound 2 is synthesized by Povarov reaction, which is then converted to compound 3 by reduction. The conversion reactions after compound 3 (deprotection of Fmoc group, N-tosylation (sulfonamidation), Suzuki-Miyaura coupling) are carried out by solid-phase synthesis. In this synthesis method, the hydroxyl group is protected by fixing it to a polymer (Si) in the conversion after compound 3. [ka]

[0004] In a method reported later (Patent Document 1), as shown in the following formula (Chemical Formula 5), ​​based on the above synthetic route, a compound corresponding to compound 3 is obtained as an N-Boc compound 3' (93% ee), and then a compound in which the hydroxy group is protected with a 3,3-dimethylbutyryl (DMB) group is used for subsequent transformations (removal of Boc, tosylation, Suzuki-Miyaura coupling, removal of DMB). In this synthetic method, the hydroxy group is protected with a DMB group and the subsequent transformations are carried out. [ka] [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication WO2020 / 068304 [Non-patent literature]

[0006] [Non-Patent Document 1] Science, 2007, 315, 1709-1712. [Non-Patent Document 2] Science, 2012, 337, 816-821. [Non-Patent Document 3] Nature Biotechnology, 2014, 32, 347-355. [Non-Patent Document 4] Cell, 2019, 177, 1067-1079.e19. [Non-Patent Document 5] ACS Comb. Sci. 2012, 14, 621-630. (Corrected in ACS Comb. Sci. 2014, 16, 46-46.) Summary of the Invention [Problem to be solved by the invention]

[0007] Using the CRISPR-Cas9 inhibitor BRD0539, it is possible to control CRISPR-Cas9 activity more precisely; therefore, if the synthesis of BRD0539 becomes easier and more readily available, this synthesis method will be highly useful. However, all of the previously reported synthetic routes to BRD0539 (Non-Patent Document 5, Patent Document 1, etc.) were carried out by protecting the hydroxy group (corresponding to the hydroxy group of the hydroxymethyl group at position 4 of the final product, BRD0539) of the starting material (compound 3 in the above chemical formula (Chemical Formula 4)). The routes involved many complicated steps, and it was believed that improvements were required. Therefore, an object of the present invention is to provide a synthetic route for BRD0539 that is more efficient than conventional synthetic routes. [Means for solving the problem]

[0008] As a result of extensive efforts, the present inventors have achieved a more efficient synthesis as shown in the following formula (Chemical Formula 6), which differs from conventional synthesis methods in that it does not require protection of the hydroxy group of the starting material (here, compound 3). [ka] First, in the first step, the amino protecting group (here, Fmoc group) of the starting material (compound 3) was deprotected, and the reaction solution was treated with amines (pyridine) and tosyl chloride (TsCl) to perform tosylation (one-pot reaction), successfully obtaining the target compound 7 in good yield (73%). Next, in the second step, the Suzuki-Miyaura coupling reaction, the reaction also proceeded in high yield (94%) without protecting the hydroxyl group, and the target compound BRD0539 was obtained. This synthetic route can obtain BRD0539 in 69% yield in two steps from compound 3, making it a very efficient synthetic method. That is, the present inventors have discovered that this reaction can be carried out without protecting the hydroxy group of the starting material, Compound A, throughout the entire reaction process, and have succeeded in developing a route to easily synthesize BRD0539 in fewer steps.

[0009] That is, the present invention provides a compound represented by the following formula (Chemical Formula 1): [ka] A method for synthesizing a compound represented by the formula: (i) Step 1: As shown in the following formula (Chemical Formula 2), a step of preparing the following compound A in a solution, deprotecting a protecting group X of an amino group of the compound A, and then reacting the following compound A with p-toluenesulfonyl chloride (TsCl) or p-toluenesulfonic anhydride in the presence of amines to synthesize a compound B; [ka] (In the formula, X represents a protecting group for an amino group, and Y represents a halogen atom or triflate (TfO). The same applies below.) (ii) second step: reacting compound B with 2-fluorophenylboronic acid or its ester, or 2-fluorophenyltrifluoroborate salt in a solution to synthesize compound BRD0539, as shown in the following formula (chemical formula 3); [ka] The method consists of the following. BEST MODE FOR CARRYING OUT THEINVENTION

[0010] The present invention is a method for synthesizing the small molecule compound BRD0539, which comprises two steps as described below.

[0011] (i) Phase 1: (a) First, prepare a solution of the following compound A. This compound A is represented by the following formula (Chemical Formula 7). [ka] In the formula, X represents an amino-protecting group.

[0012] Examples of the amino-protecting group and the deprotection method thereof are given below, however, the amino-protecting group of compound A of the present invention is not necessarily limited to these. (a) 9-Fluorenylmethyloxycarbonyl (Fmoc) group - can be deprotected by treatment with amines such as tetrabutylammonium fluoride (TBAF) or piperidine or a base. (b) tert-butoxycarbonyl (Boc) group - can be deprotected under strong acidic conditions such as trifluoroacetic acid or hydrochloric acid-ethyl acetate solution. (c) Benzyloxycarbonyl (Cbz) group - can be removed by palladium-catalyzed hydrogenation or Birch reduction. (d) 2-Nitrobenzenesulfonyl (Ns) group - can be deprotected by the action of a thiol under basic conditions. (e) 2,2,2-trichloroethoxycarbonyl (Troc) group - can be deprotected by treatment with zinc powder and acetic acid, etc. (f) Allyloxycarbonyl (Alloc) group: This can be deprotected by adding an amine in the presence of a palladium catalyst. (g) Trifluoroacetyl (CF3CO-) group -- can be removed by treatment with an aqueous solution of sodium hydroxide, etc. (h) p-Toluenesulfonyl (Ts) group - can be removed by Birch reduction, etc. (i) 2-(trimethylsilyl)ethoxycarbonyl (Teoc) group -- can be deprotected by treatment with a fluorine-containing compound (fluoride ion) such as tetrabutylammonium fluoride (TBAF).

[0013] Of these, a 9-fluorenylmethyloxycarbonyl (Fmoc) group, a tert-butoxycarbonyl (Boc) group, a benzyloxycarbonyl (Cbz) group, and a 2-nitrobenzenesulfonyl (nosyl) (Ns) group are preferred for use in the present invention. In the above formula, Y represents a halogen atom or triflate (TfO). As the solvent for the solution, an ether-based organic solvent such as THF or dioxane is preferable. The concentration of compound A in the reaction solution is preferably about 10 mM to 500 mM.

[0014] (b) Next, the protecting group X of the amino group of this compound A is deprotected. The appropriate deprotection method varies depending on the protecting group, and the method is listed above. For example, when the Fmoc group used in the examples is deprotected using TBAF, the concentration of TBAF is about 0.01 mM to 1 M. This deprotection is usually completed within 1 to several hours at a temperature of 0 to 60°C, but it is preferable to confirm that this deprotection has progressed before proceeding to the next reaction. Specifically, for example, the disappearance of the raw material is confirmed by thin layer chromatography (TLC) using UV lamp irradiation or a color developer. The extent of this deprotection is preferably such that the raw material disappears completely, but it is also acceptable if it is not complete and most of the raw material disappears.

[0015] (c) Next, p-toluenesulfonyl chloride (TsCl) or p-toluenesulfonic anhydride is added to the reaction solution of compound A having the deprotected amino group in the presence of amines to perform tosylation (sulfonamidation). Examples of the amines that can be used include pyridine, triethylamine, diisopropylethylamine, imidazole, 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), and 1,4-diazabicyclo[2.2.2]octane (DABCO). This reaction is usually carried out at about 0 to 60°C, preferably at room temperature, and the reaction time is about several hours to several days. The pressure is generally normal pressure. Other preferable reaction conditions are anhydrous conditions (under an argon or nitrogen atmosphere, with a calcium chloride tube attached, etc.). The concentrations of the amines, TsCl, and p-toluenesulfonic anhydride used in the reaction solution are each about 10 mM to 10 M, and preferably about 10 mM to 500 mM.

[0016] As a result, compound B represented by the following formula is produced. [ka]

[0017] (ii) Second Stage: In this step, compound B is reacted with 2-fluorophenylboronic acid or its ester or 2-fluorophenyltrifluoroborate salt in solution. This reaction is the so-called Suzuki-Miyaura coupling, and usually uses a palladium catalyst and a base. The reaction conditions can be the usual conditions for Suzuki-Miyaura coupling. Palladium catalysts include, but are not limited to, tetrakis(triphenylphosphine)palladium, tris(dibenzylideneacetone)dipalladium, palladium on carbon, palladium acetate, [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, dichlorobis(tricyclohexylphosphine)palladium, chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II), and the like. Also, instead of the palladium catalyst, nickel catalysts such as [1,1'-bis(diphenylphosphino)ferrocene]dichloronickel, nickel chloride, and bis(1,5-cyclooctadiene)nickel can be used. Examples of bases that can be used include, but are not limited to, sodium methoxide, sodium bicarbonate, potassium carbonate, sodium carbonate, sodium hydroxide, cesium fluoride, thallium hydroxide, and tripotassium phosphate hydrate. The concentrations of the base and catalyst in the reaction solution are each about 10 mM to 500 mM. As the solvent, an ether-based organic solvent such as THF, as well as water can be used.

[0018] The concentration of compound B in the reaction solution is preferably about 10 mM to 500 mM. As the reactant, 2-fluorophenylboronic acid or its ester, or 2-fluorophenyltrifluoroborate salt is used. Examples of the ester include pinacol ester (pin), trimethylene glycol ester, diisopropyl ester, catechol ester, neopentyl glycol ester, etc., and examples of the 2-fluorophenyltrifluoroborate salt include, but are not limited to, its potassium salt and its tetrabutylammonium salt. The concentration of these reactants in the reaction solution is about 10 mM to 500 mM. This reaction is usually carried out at about 0 to 60°C, preferably at room temperature, and the reaction time is about several hours to several days. The pressure is generally normal pressure. Other preferable reaction conditions are anhydrous conditions (under an argon or nitrogen atmosphere, with a calcium chloride tube attached, etc.). The result is the small molecule compound BRD0539.

[0019] These first and second steps may be carried out in any of the following ways: (1) After extracting and purifying compound B, which is the reaction product of the first step, the second step is carried out separately using this compound B. (2) The second step is carried out without extracting compound B, which is the reaction product of the first step. EXAMPLES

[0020] The present invention will be illustrated by the following examples, but is not intended to limit the present invention. In the following examples, melting points are uncorrected values ​​measured with a Yanaco MP-500P. NMR spectra were measured with a JEOL ECX 400 spectrometer ( 1 H-NMR: 399.8 MHz, 13 C-NMR: 100.5 MHz). 1 H and 13 The reported chemical shifts for the C NMR spectra are based on the internal standard tetramethylsilane (δ H = 0) or the solvent signal (CDCl3δ CThe IR spectra were measured using a FT IR spectrometer (Shimadzu Prestige-21). The optical rotation was measured using an AUTOPOL 4T from Rudolph Research Analytical Co., and the specific rotation was reported as 10 -1 deg cm 2 g -1 The units are reported. High-resolution mass spectra (HRMS) were measured using a mass spectrometer (Agilent Technologies 6530C qTOF) in ESI positive mode. Flash column chromatography and silica gel column chromatography were all performed using Kanto Chemical silica gel 60N (spherical, neutral, 40-50 mm). Merck Kieselgel 60 F254 was used for thin-layer chromatography.

[0021] Manufacturing Example 1 In this preparation example, (3aR,4R,9bR)-1-((9H-fluoren-9-yl)methyl) 4-ethyl 8-bromo-3,3a,4,5-tetrahydro-1H-pyrrolo[3,2-c]quinoline-1,4(2H,9bH)-dicarboxylate (compound 2) was synthesized. The reaction formula of this preparation example is shown below. [ka] Under an argon atmosphere, 2-oxoethyl acetate (Tokyo Chemical Industry Co., Ltd.) (47% toluene solution, 1.37 g, 5.53 mmol) was added to anhydrous magnesium sulfate (0.312 g) and cooled to 0°C. A solution of 4-bromoaniline (Tokyo Chemical Industry Co., Ltd.) (0.793 g, 4.61 mmol) in anhydrous toluene (Fujifilm Wako Pure Chemical Industries Co., Ltd.) (9.5 mL) was added over 30 minutes using a syringe pump, and the mixture was stirred at 0°C for 1 hour. The reaction solution was suction filtered and concentrated under reduced pressure to obtain an intermediate imine (compound 1). Toluene was added and distilled off (water was removed by azeotropy), and the subsequent reaction was then carried out. Under an argon atmosphere, compound 1 was dissolved in anhydrous toluene (27.3 mL), and the chiral urea catalyst (R)-N-[(1R,2R)-2-(3-(3,5-bis(trifluoromethyl)phenyl)ureido)cyclohexyl]-tert-butyl-sulfinamide (Sigma-Aldrich) (0.0871 mg, 0.184 mmol), powdered 5 Å molecular sieves (0.0792 g), and a solution of (9H-fluoren-9-yl)methyl 2,3-dihydro-1H-pyrrole-1-carboxylate (self-made) (1.65 g, 5.94 mmol) in anhydrous toluene (9.5 mL) were added, followed by stirring at room temperature for 5 minutes. After cooling the reaction solution to -65°C, anhydrous p-toluenesulfonic acid (0.0158 g, 0.0917 mmol) in anhydrous THF (FUJIFILM Wako Pure Chemical Industries, Ltd.) (200 μL) was added in small portions and stirred at -65°C for 1 hour. The reaction was terminated by concentrating the reaction solution under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (hexane: EtOAc = 1:1, silica gel 407 g) to obtain compound 2 (1.78 g, 70% yield).

[0022] Manufacturing Example 2 In this preparation example, (3aR,4R,9bR)-(9H-fluoren-9-yl)methyl 8-bromo-4-(hydroxymethyl)-2,3,3a,4,5,9b-hexahydro-1H-pyrrolo[3,2-c]quinoline-1-carboxylate (compound 3) was synthesized from compound 2. The reaction scheme of this preparation example is shown below. [ka] Under an argon atmosphere, compound 2 (2.12 g, 3.88 mmol) obtained in Preparation Example 1 was dissolved in anhydrous THF (FUJIFILM Wako Pure Chemical Co., Ltd.) (39 mL) and cooled to 0°C, and then LiBH4in THF (Kanto Chemical Co., Ltd.) (2.0 M, 2.9 mL, 5.8 mmol) was added, and the temperature was slowly raised to room temperature and stirred at room temperature for 3 hours. The reaction was terminated by adding sat. NH4Cl aq. (ca. 20 mL), and the mixture was extracted twice with EtOAc (ca. 15 mL). The extracted organic layer was washed with saturated saline (ca. 20 mL), dried over MgSO4, filtered by suction, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (hexane: EtOAc = 1:1, silica gel 407 g) to obtain compound 3 (1.19 g, 82% ee, yield 74%). These were recrystallized twice from benzene:CH2Cl2= 9:1 to give compound 3 (0.996 g) with 99% ee. (Chiral OJ-H column, MeOH:EtOH=1:1, 1.0 mL / min, 210 nm; retention time: timemajor = 6.64 and timeminor = 5.91 min.) Compound 3: mp 107.8~110.1℃; [α]D20.0 +135.6°(c 0.980, CHCl3); 1H-NMR (399.8 MHz, CDCl3): δ = 7.77 (d, 1 H, J = 7.3 Hz), 7.71 (d, 1 H, J = 1.8 Hz), 7.67-7.59 (m, 2 H), 7.47-7.37 (m, 3 H), 7.36-7.28 (m, 2 H), 7.10 (dd, 1 H, J = 2.3, 8.7 Hz), 6.40 (d, 1 H, J = 8.7 Hz), 5.25 (d, 0.7 H, J = 7.3 Hz), 5.09 (d, 0.3 H, J = 6.7 Hz), 4.57-4.46 (m, 1 H) 4.46-4.37 (m, 1 H), 4.37-4.18 (m, 2 H), 3.84-3.72 (m, 1 H), 3.71-3.50 (m, 2 H), 3.50-3.34 (m, 1 H) , 2.41-2.29 (m, 1 H), 2.01 (quin, 1 H, J = 11.1 Hz),1.94-1.75 (m, 2 H), 1.64 (s, 1 H) ppm; 13 C-NMR (100.5 MHz, CDCl3): δ = 156.6, 143.9,143.9, 142.1, 141.3, 132.6, 131.1, 130.9, 127.7, 127.3, 127.0, 125.0, 125.0, 124.8, 124.2, 119.9, 116.2, 110.1, 67.8, 67.5, 65.0, 56.4, 56.2, 52.7, 52.4, 47.3, 47.2, 45.1, 38.9, 38.4, 34.9, 22.6, 21.6; Anal. Calcd for C 27 H 25 BrN2O3: C, 64.16; H, 4.49; N, 5.54. HRMS (ESI+) Calcd for C 27 H 25 BrN2O3[M+H]+: 505.1127, found: 505.1126.

[0023] Example 1 In this example, BRD0539 was synthesized in two steps from compound 3. The reaction scheme of this example is shown below. [ka]

[0024] Step 1: ((3aR,4R,9bR)-8-bromo-1-tosyl-2,3,3a,4,5,9b-hexahydro-1H-pyrrolo[3,2-c]quinolin-4-yl)methanol (compound 7) was synthesized from compound 3. Under an argon atmosphere, compound 3 (1.08 g, 2.14 mmol) obtained in Production Example 2 was dissolved in anhydrous THF (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) (79.3 mL), TBAF in THF (manufactured by Sigma-Aldrich Co., Ltd.) (1.0 M, 4.3 mL, 4.3 mmol) was added, and the mixture was stirred at room temperature for 2 hours. After confirming the disappearance of the raw materials by TLC, dehydrated pyridine (manufactured by Kanto Chemical Co., Ltd.) (13.7 mL) was added, and the mixture was cooled to 0°C, and TsCl (p-toluenesulfonyl chloride (or tosyl chloride) / manufactured by Tokyo Chemical Industry Co., Ltd.) (0.612 g, 3.21 mmol) was added, and the mixture was stirred at room temperature for 3 hours. After that, TsCl (0.312 g, 1.63 mmol) was added and stirred for 15 hours, and then TsCl (0.502 g, 2.63 mmol) was added and stirred for 2 hours, after which water (ca. 100 mL) was added to stop the reaction, and after stirring for a while, the mixture was extracted three times with CHCl (ca. 30 mL). The extracted organic layer was washed with saturated brine (ca. 100 mL), dried over MgSO, suction filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash column chromatography (hexane: EtOAc = 1:3, silica gel 407 g) to obtain compound 7 (0.683 g, 73% yield). A sample for analysis was obtained by recrystallization (EtOAc). Compound 7: mp 207.2~208.5℃; [α]D20.0 +103°(c 1.07, CHCl3); 1H-NMR (CDCl3 / TMS): δ = 7.82 (s, 1H), 7.78 (d, 2H, J = 8.2 Hz), 7.36 (d, 2H, J = 7.8 Hz), 7.12 (d, 1H, J = 8.7 Hz), 6.39 (dd, 1H, J = 1.1, 8.7). Hz), 5.00 (d, 1 H, J = 7.3 Hz), 4.19 (brs, 1 H), 3.63 (d, 1 H, J = 7.3 Hz), 3.55–3.44 (m, 2 H), 3.34–3.36 (m, 1 H), 2.46 (s, 3 H), 1.92 (quin, 1 H, J). = 9.7 Hz), 1.77–1.63 (m, 2 H) ppm; 13 C-NMR (CDCl3 / TMS): δ = 143.8, 143.2, 135.0, 133.4, 131.1, 129.9, 127.6, 122.4, 116.2, 110.5, 65.0, 58.9, 52.5, 47.2, 38.3. 23.1, 21.6; Anal. Calcd for C 19 H 21 BrN2O3S: C, 52.18; H, 4.89; N, 6.41. Found: C, 52.04; H, 4.82; N, 6.31%.

[0025] Compound 2: The active compound 7 ((3aR,4R,9bR)-8-(2-fluorophenyl)-1-tosyl-2,3,3a,4,5,9b-hexahydro-1H-pyrrolo[3,2-c]quinolin-4-yl)methanol (BRD0539) . Under an argon atmosphere, compound 7 (0.683 g, 1.56 mmol) obtained in the first step was dissolved in THF (52.0 mL) and distilled water (17.3 mL) was added. Then, 2-fluorophenylboronic acid (Tokyo Chemical Industry Co., Ltd.) (0.363 g, 2.34 mmol), K3PO4·nH2O (tripotassium phosphate hydrate 76.7%, 1.12 g, 4.05 mmol), and palladium catalyst (chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) / Sigma-Aldrich X-Phos Pd-G2) (0.0614 g, 0.0781 mmol) were added and the mixture was stirred at room temperature for 3 hours. The reaction mixture was extracted three times with EtOAc (ca. 20 mL), and the organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to give the crude product. The crude product was purified by flash column chromatography (hexane: EtOAc = 1:2, silica gel 212 g) to give BRD0539 (0.661 g, 94% yield). An analytical sample was obtained by recrystallization (MeOH). BRD0539: mp 101.2.0~104.4℃; [α]D20.0 105°(c 0.960, CHCl3); 1 H-NMR (CDCl3 / TMS): δ = 7.90 (s, 1 H), 7.79 (d, 2 H, J = 7.8 Hz), 7.43 (t, 1 H, J = 7.8 Hz), 7.36-7.27 (m, 3 H), 7.25-7.04 (m, 3 H), 6.56 (d, 1 H, J = 8.2 Hz), 5.09 (d, 1 H, J = 7.3 Hz), 4.28 (brs, 1 H ), 3.62 (dd, 3 H, J =2.3, 9.2 Hz), 3.58-3.42 (m, 2 H), 3.42-3.25 (m, 2 H), 2.43 (s, 3 H), 1.98 (quin, 1 H, J = 10.6 Hz), 1.87-1.73 (m, 1 H), 1.71-1.57, (m, 2 H) ppm; 13C-NMR (CDCl3 / TMS): δ = 159.7 (d, J = 246.4 Hz), 143.6, 142.9, 135.2, 131.3 (d, J = 1.9 Hz),130.4 (d, J = 3.8 Hz), 129.9, 129.0 (d, J = 4.8 Hz), 127.9 (d, J = 7.6 Hz), 127.6, 126.2,124.3 (d, J = 3.8 Hz), 120.5, 116.0, 115.8, 114.5, 65.1, 59.2, 52.6, 47.2, 38.7, 23.2, 21.5; Anal. Calcd for C 27 H 25 BrN2O3: C, 66.35; H, 5.57; N, 6.19. Found: C, 66.34; H, 5.73; N, 6.15 %.

Claims

1. The following formula (chemical formula 1) 【Chemistry 1】 A method for synthesizing a compound represented by the formula: (i) First step: as shown in the following formula (Chemical Formula 2), a step of preparing the following compound A in a solution, deprotecting a protecting group X of an amino group of the compound A, and then reacting the following compound A with p-toluenesulfonyl chloride (TsCl) or p-toluenesulfonic anhydride in the presence of amines to synthesize a compound B; 【Chemistry 2】 (In the formula, X represents a protecting group for an amino group, and Y represents a halogen atom or triflate (TfO). The same applies below.) (ii) second step: reacting compound B with 2-fluorophenylboronic acid or its ester, or 2-fluorophenyltrifluoroborate salt in a solution to synthesize compound BRD0539, as shown in the following formula (chemical formula 3); 【Chemistry 3】 The method comprising:

2. 2. The method of claim 1, wherein the first and second steps are carried out in one of the following ways: (1) After extracting and purifying the compound B, which is the reaction product of the first step, the second step is carried out separately using this compound B. (2) The second step is carried out without extracting compound B, which is the reaction product of the first step.

3. 3. The method according to claim 1, wherein the amino-protecting group X is a 9-fluorenylmethyloxycarbonyl (Fmoc) group, a tert-butoxycarbonyl (Boc) group, a benzyloxycarbonyl (Cbz) group, or a 2-nitrobenzenesulfonyl (nosyl) (Ns) group.

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