AU2020101059A4 - Preparation method of indoleamine 2,3-dioxygenase inhibitor - Google Patents

Abstract

The present invention relates to a preparation method for compounds shown in formula I. The compounds prepared by the method of the present invention have activity of inhibiting indoleamine 2,3-dioxygenase. [Ri] R2 Norl (1N-Cy2 N Cy1 -(CRaRb) A formula (I)

Description

PREPARATION METHOD OF INDOLEAMINE 2,3-DIOXYGENASE INHIBITOR

TECHNICAL FIELD

[0001] The present invention relates to the field of drugs, and particularly relates to a preparation method of an indoleamine 2,3-dioxygenase inhibitor.

BACKGROUND OF THE PRESENT INVENTION

[0002] Tryptophan (TRP) is an a-amino acid used for protein biosynthesis. TRP contains a-amino groups, a-carboxylic acid groups and side-chain indole. TRP is indispensable to human beings, but cannot be synthesized by the human body, and can be obtained only from the diet. TRP is also a precursor for synthesizing a neurotransmitter 5-hydroxytryptamine (serotonin) and hormone N-acetyl-5-methoxytryptamine (melatonin). The heme-dependent enzyme indoleamine 2,3-dioxygenase (also called IDO, or IDOl) is an extrahepatic metabolic enzyme that is responsible for converting the tryptophan to N-formyl-kynurenine, which is the first step of a metabolism process of the tryptophan and is also the rate-limiting step of the entire process. N-formyl-kynurenine is a precursor of various bioactive molecules of kynurenine (Kyn), and kynurenine has an immunomodulatory function (Schwarcz et al, Nat Rev Neurosci. 2012; 13(7): 465).

[0003] The research on IDO inhibitor anti-tumor drugs has already made a great progress in the world, for example, INCB024360, NLG919 and BMS-986205 have all entered the clinical phase. However, INCB024360 has problems of toxicity and side effects, resulting in that the existing clinical research dose (50mg bid or 100mg bid) is about 30% of the optimum dose (300mg bid, 600mg bid), and the clinical activity is greatly limited. Meanwhile, toxic groups of the INCB024360 are pharmacophores, and INCB024360 and derivatives thereof have a problem of greater toxicity. NLG919 is good in safety, but is poor in biological activity. BMS-986205 has also already entered the clinical phase at present, but the clinical data is limited. Based on BMS-986205, the research on novel compounds with high biological activity and high safety has important practical significance in discovering novel IDO tumor immunotherapy drugs with better clinical treatment activity such as the possibility of treating the tumor rather than merely inhibiting the tumor.

SUMMARY OF PRESENT INVENTION

[0004] The present invention also provides a preparation method for compounds having structure of formula (X):

[R1] 0 [R1] OEt c Rd

N EcY N0 ylides NODt N-, base/organic solvent N step (1)

R2 R Rd R2 HN-Cy 2 N-CY2 Grignard reagents

Formula (X) route I

[0005] wherein ^^^ represents: - , nu or m.-;

[0006] wherein each R is independently selected from a hydrogen atom, halogen, hydroxyl, nitro, cyano, sulfonate, C1-6 alkyl, C3 -6cycloalkyl, C 2 -6 alkenyl, C 2 -6 alkynyl, C 1 .6 alkoxy, and halogenated C1 -C 6 alkyl;

[0007] wherein R2 is independently selected from hydrogen, C1 -C 6 alkyl or C3-6 cycloalkyl;

[0008] wherein Rc and Rd are independently selected from hydrogen or C1.6 alkyl;

[0009] wherein Cy2 is C 5 -C 1 0 aryl and C 5 -C 1 0 heteroaryl containing one or more than one substituent group;

[0010] wherein m is an integer of 0-4.

[0011] The base used instep (1) is selected from inorganic bases or organic bases, including but not limited to: sodium hydride, calcium hydride, sodium amide, sodium methoxide, sodium ethoxide, potassium hydroxide, sodium hydroxide, lithium hydroxide, lithium aluminium hydride, tert-butyl lithium, tert-butyl potassium, potassium tert-butoxide, lithium diisopropylamide, barium hydroxide, or any combination thereof.

[0012] The organic solvents used instep (1) include but are not limited to: 1,4-dioxane, N,N-dimethylformamide, dichloromethane, chloroform, DMSO, DMF, THF, acetone, methanol, ethanol or any combination thereof.

[0013] The ylides used instep (2) are selected from sulfur ylide or phosphorus ylide.

[0014] The Grignard reagents used instep (3) are selected from CH3MgCl, CH 3MgBr, C 2 H 5MgCl, C 2H 5MgBr, i-PrMgCl, i-PrMgBr, PhCH 2 MgCl, PhCH 2MgBr or any combination thereof.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[0015] Example 1

[0016] N-(4-chlorophenyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-carboxami de F H N

N /

CI FC1O O PhNTf 2 O PddfCF NaHMDS KOAc, NaBr O O O

MTBE TfO 1,4-dioxane Pd(PPh 1,4-dioxane/H 2 0 N

la lb 1c

F F 0F

0O COOEt 0C Pd/C,H 2 N) 1-P ~ 4N____ IPA N ceon N - 0 t-BuONa. THF N I

1d le if

oF F H F O OEt 4-chloroaniline N OC

NaH, DMSO i-PrMgCI, THF

1g 1

[0017] Step 1: To a solution of 1, 4-cyclohexanedione monoethylene acetal (10.0 g, 64.03 mmol) in tert-butyl methyl ether (250 mL) was added N-phenyl-bis(trifluoromethanesulfonimide (22.9 g, 64.03 mmol). The mixture was cooled to -78°C, and sodium bis(trimethylsilyl)amide (2.0 M in THF, 32 mL, 64.03 mmol) was added dropwise under N 2 . The mixture was stirred at -78°C for 1 h and then warmed to r.t. for 16 h until TLC showed complete conversion of starting material. The reaction was quenched by 3 mL KHS04 solution and filtered. The filtrate was concentrated under reduced pressure. The residue was dissolved in 30 mL MTBE, which was washed with 5% NaOH (45 mL x 3) and brine (50 mL). The organic layer was dried over Na 2 SO 4 , filtered and concentrated to give compound la

(34 g, yield: 93.6%) as red-orange oil. 'H NMR (500 MHz, CDCl3) 6 5.66 (J= 4.0 Hz, 1H), 4.01-3.96 (m, 4H), 2.56-2.52 (m, 2H), 2.42-2.40 (m, 2H), 1.90 (t, J= 6.5 Hz, 2H).

[0018] Step 2: To a solution of compound la (13 g, 45.1 mmol) in dioxane (100 mL) were added bis(pinacolato)diboron (14.9 g, 58.64 mmol), KOAc (13.3 g, 135.3 mmol) and Pd(dppf)C12 (1.65 g, 2.26 mmol). The reaction mixture was stirred at 100°C under

N 2 for 16 h. The mixture was concentrated under reduced pressure. The residue was suspended in EA and filtered over a pad of celite. The filtrate was concentrated and purified by column chromatography on silica gel to afford compound lb (7.6 g, yield: 63%) as a pale-yellow solid. IH NMR (500 MHz, CDCl 3) 6 6.48-6.45 (m, 1H), 3.98 (s, 4H), 2.40-2.34 (m, 4H), 1.73 (t, J= 6.5 Hz, 2H), 1.25 (s, 12H).

[0019] Step 3: To a solution of compound lb (5.7 g, 21.48 mmol) in dioxane (60 mL) and water (15 mL) were added 4-chloro-6-fluoroquinoline (3.0 g, 16.53 mmol), potassium carbonate (6.8 g, 49.56 mmol), and tetrakis(triphenylphosphine) palladium(0) (954 mg, 0.83mmol). The reaction mixture was stirred at 100°C under N 2 for 16 h. The mixture was concentrated under reduced pressure. The residue was diluted with H 2 0 and extracted with EtOAc. The organic layer was concentrated. The residue was purified by column chromatography on silica gel to afford compound le (2.42 g, yield 51%) as pale-yellow oil. MS (ESI): m/z 286.1 (M+H). 'H NMR (500 MHz, CDC 3) 68.81 (d, J= 4.5 Hz, 1H), 8.15 (dd, J= 9.0, 5.5 Hz, 1H), 7.65 (dd, J= 10.0, 2.5 Hz, 1H), 7.49 (td, J= 9.0, 2.5 Hz, 1H), 7.26 (d, J= 4.5 Hz, 1H), 5.77 (t, J 3.5 Hz, 1H), 4.08-40.6 (m, 4H), 2.65-2.60 (m, 2H), 2.56-2.53 (m, 2H), 2.00 (t, J= 6.5 Hz, 2H).

[0020] Step 4: To a solution of compound le (2.42 g, 8.49 mmol) in isopropyl alcohol (45 mL) was added 10% w/w palladium on carbon (300 mg). The mixture was stirred at 55°C under hydrogen atmosphere for 16 h. The reaction was filtered over a pad of celite. The filtrate was concentrated to give compound id (2.04 g, yield: 84%) as slurry oil, which was used to the next step directly. MS (ESI): mlz 288.1 (M+H).

[0021] Step 5: To a solution of compound ld (2.04 g, 7.11 mmol) in acetone (36 mL) was added HCl (4N in water, 9 mL, 36 mmol) and the mixture was stirred at 45°C for 16 h. The organic volatiles were removed and the remaining aqueous solution was adjusted to pH=9 by 6N NaOH solution. The mixture was extracted with EtOAc three times. The combined organic layers were washed with brine, dried over Na2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to give compound le (1.17 g, yield 67%) as a pale-yellow solid. MS (ESI): m/z 244.3 (M+H).'H NMR (500 MHz, CDCl 3) 6 8.85 (d, J= 4.5 Hz, 1H), 8.22 (dd, J= 9.0, 5.5 Hz, 1H), 7.74 (dd, J= 10.0, 2.5 Hz, 1H), 7.57-7.50 (m, 1H), 7.33 (d, J= 4.5 Hz, 1H), 3.74-3.66 (m, 1H), 2.72-2.58 (m, 4H), 2.41-2.34 (m, 2H), 2.11-2.00 (m, 2H).

[0022] Step 6: To a solution of triethyl phosphonoacetate (968 mg, 4.32 mmol) in dry THF (16 mL) at0°C, sodium tert-butoxide (415 mg, 4.32 mmol) was added. After min, a solution of compound le (1 g, 4.12 mmol) in THF (4 mL) was added to the above mixture and the resulting reaction mixture was stirred for 2 h. The reaction was quenched by H 20, extracted with EtOAc three times. The combined organic layers were washed with brine (20 mL), dried over Na2 SO4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel to give compound 1f (1.18 g, yield: 92%) as a white solid. MS (ESI): mlz 314.0 (M+H). H NMR (500 MHz, CDC 3) 68.81 (d, J= 4.5 Hz, 1H), 8.17 (dd, J= 9.0, 5.5 Hz, 1H), 7.72 (dd, J= 10.0, 2.5 Hz, 1H), 7.53-7.47 (m, 1H), 7.28 (d, J= 4.5 Hz, 1H), 5.75 (s, 1H), 4.19 (q, J= 7.0 Hz, 2H), 3.52-3.42 (m, 1H), 2.54-2.48 (m, 2H), 2.26-2.11 (m, 4H), 1.80-1.68 (m, 2H), 1.30 (t, J= 7.0 Hz, 3H).

[0023] Step 7: To a suspension of NaH (60% w/w in mineral oil, 383 mg, 9.57 mmol) in DMSO (15 mL) was added trimethylsulfoxonium iodide (2.11 g, 9.57 mmol). After the mixture was stirred at r.t. for 1.5 h, a solution of compound 1f (1.0 g, 3.19 mmol) in DMSO (5 mL) was added to the above mixture and the resulting reaction mixture was stirred at r.t. for 16 h. The mixture was quenched by H 2 0 and extracted with EtOAc. The organic layer was concentrated and the residue was purified by column chromatography on silica gel to give compound 1g (820 mg, yield: 78%) as colorless oil. MS (ESI): m/z 328.1 (M+H)F. 'H NMR (500 MHz, CDC 3) 6 8.83 (d, J= 4.5 Hz, 1H), 8.24 (dd, J= 9.0, 5.5 Hz, 1H), 7.71 (dd, J= 10.0, 2.5 Hz,1H), 7.55-7.49 (m,1H), 7.35 (d, J= 4.5 Hz, 1H), 4.19 (q, J= 7.0 Hz, 2H), 3.32-3.24 (m, 1H), 2.17 (td, J= 13.0, 3.5 Hz, 1H), 2.07-1.90 (m, 4H), 1.87-1.78 (m, 1H), 1.58 (dd, J= 8.0, 5.5 Hz, 1H), 1.46-1.37 (m, 1H), 1.30 (t, J= 7.0 Hz, 3H), 1.28-1.24 (m, 2H), 1.16-1.11 (m, 1H), 1.00 (dd, J= 8.0, 4.5 Hz, 1H).

[0024] Step 8: To a mixture of 4-chloroaniline (94 mg, 0.73 mmol) in dry THF (5 mL) at 0°C, isopropylmagnesium chloride solution (2.0 M in THF, 0.4 mL, 0.73 mmol) was added dropwise. After the mixture was stirred at r.t. for 5 min, a solution of compound 1g (60 mg, 0.l8mmol) in dry THF (2 mL) was added dropwise. The resulting mixture was stirred at r.t. for 16 h. The reaction was quenched by aq. NH 4 Cl solution. The mixture was extracted with ethyl acetate three times and the combined organic layers were dried over Na2 SO4 , filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give compound 1 (16.04 mg, yield: 21%) as a white solid. MS (ESI): m/z 408.9 (M+H).'H NMR (500 MHz, d6 -DMSO) 6 10.37 (s, 1H), 8.81 (s,1H), 8.11-8.05 (m, 1H), 8.02 (d, J= 11.0 Hz, 1H), 7.69-7.63 (m, 3H), 7.38-7.31 (m, 3H), 3.48-3.40 (m, 1H), 2.20 (t, J= 12.0 Hz,1H), 1.97-1.84 (m, 4H), 1.78 (d, J= 12.5 Hz, 1H), 1.72 (t, J= 6.5 Hz, 1H), 1.35-1.26 (m, 1H), 1.17-1.08 (m, 2H), 0.96-0.90 (m, 1H).

[0025] Example 2

[0026] N-(4-fluorophenyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-carboxam ide F H

NN F N /

[0027] Compound 2 was prepared using the similar procedures as described for compound 1 using 4-fluoro aniline to replace 4-chloro aniline. MS (ESI): mlz 393.3 (M+H)f.'H NMR (500 MHz, d6 -DMSO) 6 10.28 (s, 1H), 8.81 (s, 1H), 8.10-8.05 (m, 1H), 8.03 (d, J= 11.0 Hz, 1H), 7.69-7.60 (m, 3H), 7.37 (s, 1H), 7.13 (t, J= 8.0 Hz, 2H), 3.49-3.40 (m, 1H), 2.20 (t, J= 12.0 Hz, 1H), 1.98-1.85 (m, 4H), 1.78 (d, J= 11.0 Hz, 1H), 1.72 (t, J= 6.5 Hz, 1H), 1.37-1.28 (m, 1H), 1.17-1.07 (m, 2H), 0.94-0.89 (m, 1H).

[0028] Example 3

[0029] N-(4-chlorobenzyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-carboxam ide CI F H ::

1 0 N /

FF F CI

N OH HATU, DIPEA OEt E OH

1g 3a 3

[0030] Step 1: To a solution of compound 1g (200 mg, 0.61 mmol) in ethanol (10 mL) was added NaOH (2N in water, 4 mL, 8 mmol) and the mixture was stirred at °C for 2 h. The mixture was cooled to r.t. and adjusted to pH=1 by 4N HCl solution. The mixture was extracted with EtOAc three times. The combined organic layers were dried over Na2 SO4 ,filtered and concentrated under reduced pressure. The residue was purified by Prep-TLC to give compound 3a (150 mg, yield 83%) as a white solid. MS (ESI): m/z 300.0 (M+H). 1H NMR (500 MHz, d6-DMSO) 612.02 (br, 1H), 8.83 (d, J = 4.5 Hz, 1H), 8.10 (dd, J= 9.0, 5.5 Hz, 1H), 8.03 (dd, J= 10.0, 2.5 Hz, 1H), 7.71-7.64 (m, 1H), 7.38 (d, J= 4.5 Hz, 1H), 3.48-3.41 (m, 1H), 2.21-2.13 (m, 1H), 2.01-1.80 (m, 4H), 1.75-1.65 (m, 1H), 1.51 (dd, J= 8.0, 5.5 Hz, 1H), 1.38-1.32 (m, 1H), 1.11-1.05 (m, 1H), 1.04-0.99 (m, 1H), 0.95 (dd, J= 7.5, 4.0 Hz, 1H).

[0031] Step 2: To a solution of compound 3a (40 mg, 0.13 mmol) inDMF (5 mL) were added DIPEA (52 mg, 0.39 mmol) and HATU (61 mg, 0.16 mmol) and the mixture was stirred at r.t. for 30 min. 4-chlorobenzylamine (57 mg, 0.39 mmol) was added to the above mixture and the resulting reaction mixture was stirred at r.t. for 2 h. The mixture was quenched by H2 0(20 mL) and extracted with EtOAc three times. The combined organic layers were dried over Na2 SO4 ,filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give compound 3 (6.34 mg, yield 11%) as a white solid.MS (ESI): m/z 423.4 (M+H)+. 'H NMR (500 MHz, d6 -DMSO) 6 8.81 (s, 1H), 8.68 (s, 1H), 8.12-8.07 (m, 1H), 8.00 (d, J= 11.0 Hz, 1H), 7.67 (t, J= 8.5 Hz, 1H), 7.36-7.31 (m, 4H), 7.14 (s, 1H), 4.44 (dd, J= 15.0, 6.5 Hz, 1H), 4.18 (dd, J= 15.0 Hz, 5.0 Hz,1H), 3.42-3.34 (m, 1H), 2.13 (t, J= 12.5 Hz, 1H), 1.86-1.74 (m, 4H), 1.63 (d, J= 12.5 Hz, 1H),1.54-1.48 (m, 1H), 1.25-1.15 (m, 1H), 1.08-0.96 (m, 2H), 0.82-0.76 (m, 1H).

[0032] Example 4

[0033] N-(4-chlorophenethyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-carbo xamide

F H N

0

N /

[0034] Compound 4 was prepared using the similar procedures as described for compound 3 using 4-chlorophenethylamine to replace 4-chlorobenzylamine. MS (ESI): m/z 437.4 (M+H)f.'H NMR (500 MHz, d6 -DMSO) 6 8.84 (d, J= 4.5 Hz, 1H), 8.19 (t, J= 5.5 Hz, 1H), 8.12-8.07 (m, 1H), 7.99 (d, J= 11.0 Hz, 1H), 7.67 (t, J= 8.5 Hz, 1H), 7.27 (d, J= 4.1 Hz,1H), 7.25-7.21 (m, 4H), 3.49-3.42 (m, 1H), 3.30-3.22 (m, 2H), 2.74 (t, J= 6.5 Hz, 2H), 2.11 (t, J= 12.5 Hz,1H), 1.85-1.75 (m, 4H), 1.67 (d, J= 12.5 Hz, 1H), 1.46-1.41 (m, 1H), 1.26-1.14 (m, 1H), 1.03-0.95 (m, 2H), 0.75-0.70 (m, 1H).

[0035] Example 5

[0036] 6-(6-fluoroquinolin-4-yl)-N-(4-(trifluoromethyl)phenyl)spiro[2.5]octane 1-carboxamide F H

00N 1KCF 3 N /

F F H II:OH N T3P, Pyridine N CF 3 N N

3a 5

[0037] To asolutionof compound3a (40 mg, 0.13 mmol) inEA (5 mL) were subsequently added pyridine (32 mg, 0.39 mmol) and 2,4,6-Tripropyl-1,3,5,2,4,6-Trioxatriphosphorinane-2,4,6-Trioxide (127 mg, 0.33 mmol) and the mixture was stirred at r.t. for 10 min. 4-Aminobenzotrifluoride (65 mg, 0.39 mmol) was added to the above mixture and the resulting reaction mixture was stirred at r.t. for 16 h. The mixture was quenched by NaOH (2N in water, 2 mL) and diluted with H 2 0 (20 mL). The resulting mixture was extracted with EtOAc three times. The combined organic layers were dried over Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give compound 5 (2.65 mg, yield 5%) as a white solid. MS (ESI): m/z 437.4 (M+H). IH NMR (500 MHz, d-DMSO) 6 10.63 (s, 1H), 8.80 (d, J= 4.5 Hz, 1H), 8.10-8.05 (m, 1H), 8.03 (d, J= 11.0 Hz, 1H), 7.84 (d, J= 8.0 Hz, 2H), 7.69-7.63 (m, 3H), 7.36 (s, 1H), 3.48-3.41 (m, 1H), 2.25-2.17 (m, 1H), 1.99-1.85 (m, 4H), 1.80-1.74 (m, 2H), 1.35-1.25 (m, 1H), 1.21-1.09 (m, 2H), 0.99-0.95 (m, 1H).

[0038] Example 6

[0039] N-(4-cyanophenyl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-carboxam ide F H

C N /

[0040] Compound 6 was prepared using the similar procedures as described for compound 5 using 4-aminobenzonitrile to replace 4-aminobenzotrifluoride. MS (ESI): m/z 400.4 (M+H).'H NMR (500 MHz, d 6-DMSO) 6 10.70 (s, 1H), 8.80 (d, J= 4.5 Hz, 1H), 8.10-8.06 (m, 1H), 8.03 (d, J= 11.0 Hz, 1H), 7.82 (d, J= 8.5 Hz, 2H), 7.75 (d, J= 8.5 Hz, 2H), 7.66 (t, J= 9.0 Hz, 1H), 7.35 (d, J= 3.5 Hz, 1H), 3.48-3.40 (m, 1H), 2.25-2.17 (m, 1H), 1.99-1.83 (m, 4H), 1.81-1.74 (m, 2H), 1.33-1.23 (m, 1H), 1.21-1.10 (m, 2H), 1.02-0.96 (m, 1H).

[0041] Example 7

[0042] N-(6-chloropyridin-3-yl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-1-car boxamide F H N

CI N /

[0043] Compound 7 was prepared using the similar procedures as described for compound 5 using 5-amino-2-chloropyridine to replace 4-aminobenzotrifluoride. MS (ESI): m/z 410.4 (M+H)f.'H NMR (500 MHz, d6 -DMSO) 6 10.62 (s, 1H), 8.81 (s, 1H), 8.63 (s, 1H), 8.15-8.06 (m, 2H), 8.03 (d, J= 11.0 Hz, 1H), 7.66 (t, J= 8.5 Hz, 1H), 7.45 (d, J= 8.0 Hz, 1H), 7.38 (s, 1H), 3.48-3.41 (m, 1H), 2.25-2.16 (m, 1H), 1.99-1.82 (m, 4H), 1.82-1.72 (m, 2H), 1.35-1.26 (m, 1H), 1.20-1.09 (m, 2H), 1.00-0.95 (m, 1H).

[0044] Example 8

[0045] N-(5-chloropyridin-2-yl)-6-(6-fluoroquinolin-4-yl)spiro[2.5]octane-I-car boxamide F H

IN NN 0N CI N /

[0046] Compound 8 was prepared using the similar procedures as described for compound 5 using 2-amino-5-chloropyridine to replace 4-aminobenzotrifluoride. MS (ESI): m/z 410.4 (M+H)f.'H NMR (500 MHz, d6 -DMSO) 6 11.04 (s, 1H), 8.80 (s, 1H), 8.38 (s, 1H), 8.13 (d, J= 8.5 Hz, 1H), 8.07 (t, J= 8.0 Hz, 1H), 8.02 (d, J= 11.0 Hz, 1H), 7.85 (d, J= 9.0 Hz, 1H), 7.66 (t, J= 8.0 Hz,1H), 7.34 (s,1H), 3.47-3.40 (m, 1H), 2.19 (t, J= 12.5 Hz, 1H), 2.02-1.84 (m, 5H), 1.78 (d, J= 11.0 Hz, 1H), 1.33-1.22 (m, 1H), 1.21-1.15 (m, 1H), 1.08 (d, J= 12.5 Hz, 1H), 0.98-0.92 (m, 1H).

[0047] Example 9

[0048] N-(4-chlorophenyl)-6-(6-fluoro-7-methylquinolin-4-yl)spiro[2.5]octane-1 carboxamide

F > H Me N

CI N/

F F F H OEt LIDA, Mel Me OEt Prg F Me,,, i-PrMgCI, THF IC C LDe

Ig 9a 9

[0049] Step 1: To a mixture of diisopropylamine (123 mg, 1.22 mmol) in dry THF (5 mL) at -78°C, n-BuLi (2.5 M, 0.5 mL, 1.25 mmol) was added dropwise, followed by a solution of compound 1g (200 mg, 0.61 mmol) in THF (2 mL). After the mixture was stirred at -78°C for 1 h, a solution of CH 3I (173 mg, 1.22 mmol) in dry THF (2 mL) was added dropwise. The resulting mixture was stirred at -78°C for 0.5 h and then warmed to r.t. for 16 h. The reaction was quenched by aq. NH 4 Cl solution. The mixture was extracted with ethyl acetate three times and the combined organic layers were dried over Na2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by Prep-TLC to give compound 9a (24 mg, yield: 12%) as colorless oil. MS (ESI): m/z 342.4 (M+H)f.

[0050] Step 2: To a mixture of 4-chloroaniline (36 mg, 0.28 mmol) in dry THF (2 mL) at 0°C, isopropylmagnesium chloride solution (2.0 M in THF, 0.2 mL, 0.4 mmol) was added dropwise. After the mixture was stirred at r.t. for 5 min, a solution of compound 9a (24 mg, 0.07 mmol) in dry THF (1 mL) was added dropwise. The resulting mixture was stirred at r.t. for 16 h. The reaction was quenched by aq. NH 4 Cl solution. The mixture was extracted with ethyl acetate three times and the combined organic layers were dried over Na2 SO4 , filtered and concentrated under reduced pressure. The residue was purified by Prep-HPLC to give compound 9 (12.01 mg, yield: 41%) as a white solid. MS (ESI): m/z 423.4 (M+H).'H NMR (500 MHz, d6 -DMSO) 6 10.37 (s, 1H), 8.75 (s, 1H), 7.99-7.91 (m, 2H), 7.80-7.56 (m, 3H), 7.34 (d, J= 8.0 Hz, 2H), 7.28 (s, 1H), 3.44-3.37 (m, 1H), 2.44 (s, 3H), 2.23-2.14 (m, 1H), 1.98-1.84 (m, 4H), 1.78-1.69 (m, 2H), 1.17-1.07 (m, 2H), 0.95-0.92 (m, 1H).

Claims (5)

1. We claim: 1. A preparation method for compounds shown in formula (X), comprising the following reaction steps:

   [R1] 0 [R1m Et c Rd EtO COOEt [R1]m O t Fzdylides " OEt - base/organic solvent N -

   step (1) O

   R Rd R2 R2 HN-Cy 2 N-Cy 2 Grignard reagents N Formula (X) route I

   wherein ^^^ represents: - , -- or m.-; wherein each Rl is independently selected from a hydrogen atom, halogen, hydroxyl, nitro, cyano, sulfonate, C1 -6 alkyl, C 3 -6cycloalkyl, C2 -6alkenyl, C 2 -6 alkynyl, CI-6 alkoxy, and halogenated C1 -C 6 alkyl; wherein R2 is independently selected from hydrogen, C1 -C6 alkyl or C 3 -6 cycloalkyl; wherein R and Rd are independently selected from hydrogen or C1 -6alkyl; wherein Cy 2 is C 5 -C 1 0 aryl and C 5 -C 1 0 heteroaryl comprising one or more than one substituent group; wherein m is an integer of 0-4.
2. 2. The method according to claim 1, wherein the base used in step (1) is selected from inorganic bases or organic bases, comprising but not limited to: sodium hydride, calcium hydride, sodium amide, sodium methoxide, sodium ethoxide, potassium hydroxide, sodium hydroxide, lithium hydroxide, lithium aluminium hydride, tert-butyl lithium, tert-butyl potassium, potassium tert-butoxide, lithium diisopropylamide, barium hydroxide, or any combination thereof.
3. 3. The method according to claim 1 or 2, wherein the organic solvents used in step (1) comprise but are not limited to: 1,4-dioxane, N,N-dimethylformamide, dichloromethane, chloroform, DMSO, DMF, THF, acetone, methanol, ethanol or any combination thereof.
4. 4. The method according to any one of claims 1-3, wherein the ylides used in step (2) are selected from sulfur ylide or phosphorus ylide.
5. 5. The method according to any one of claims 1-4, wherein the Grignard reagents used in step (3) are selected from CH 3MgCl, CH 3MgBr, C 2 HMgCl, C 2H 5MgBr, i-PrMgCl, i-PrMgBr,

   PhCH 2 MgCl, PhCH 2MgBr or any combination thereof.