DNA polymerase theta inhibitor and application thereof

Abstract

The invention discloses a DNA polymerase theta inhibitor and application thereof, the compound has the following structure (I), and the compound has a good inhibition effect on P01 theta.

Description

DNA polymerase theta inhibitor and its application Technical Field

[0001] The present invention relates to a DNA polymerase theta inhibitor and application thereof. Background Art

[0002] Mammalian cells have evolved multiple pathways to repair DNA double-strand breaks (DSBs) to ensure genomic stability. DNA polymerase theta (Polθ) is a key component of the alternative end joining (alt-NHEJ) pathway, also known as the microhomology-mediated end joining (MMEJ) pathway, which is involved in DNA double-strand break repair. MMEJ is another repair pathway in cells in addition to non-homologous end joining (NHEJ) and homologous recombination (HR). MMEJ repair is driven by annealing of microhomology sequences flanking the DNA ends and was initially thought to be a backup pathway for gene repair. However, as research progressed, it was discovered that MMEJ is not a backup repair mechanism. Genetic, cell biological, and biochemical data have identified Polθ as a key protein in MMEJ.

[0003] When DNA end resection occurs, in the presence of BRCA2, BRCA2 not only recruits the recombinase RAD51 to the DSB to promote HR but also inhibits repair pathways such as alt-NHEJ. When homologous recombination-mediated repair is impaired (HR deficiency), as in the case of BRCA1 or BRCA2 mutations, Pol θ is highly expressed and directs DSB repair toward alt-EJ, initiating the DNA repair process of microhomology-mediated end joining (MMEJ). In the presence of HR deficiency, inhibition of Pol θ leads to cell death through the accumulation of toxic RAD51 intermediates and inhibition of the alt-EJ repair pathway.

[0004] Polθ is a multifunctional enzyme consisting of an N-terminal helicase domain and a C-terminal low-fidelity DNA polymerase domain. These two domains have been shown to have coordinated mechanistic functions in MMEJ. The helicase domain mediates the removal of RPA proteins from ssDNA ends and stimulates annealing. The polymerase domain extends the ssDNA ends and fills the remaining gaps. Polθ also reverse-transcribes RNA and promotes RNA-templated DNA repair. Polθ is barely expressed in normal tissues but is highly expressed in various tumor types (e.g., breast cancer, ovarian cancer, HNSCC, and lung cancer), and its overexpression is associated with poor prognosis. Furthermore, synthetic lethal interactions have been identified between Polθ and multiple DNA repair genes, including factors involved in homologous recombination (e.g., BRCA1 / 2). For these reasons, Polθ inhibitors represent a promising cancer treatment strategy.

[0005] Summary of the Invention

[0006] The technical problem to be solved by the present invention is that the existing Polθ inhibitors have a relatively simple structure. To this end, the present invention provides a DNA polymerase theta inhibitor and its application, which has a good inhibitory effect on Polθ.

[0007] Specifically, the present invention provides a compound having a structure of formula (I), its pharmaceutically acceptable salts, isotopic isomers, stereoisomers,

[0008] Among them, R 1 Indicates -LR L ;

[0009] Among them, R 2 represents hydrogen, deuterium, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、CN、nitro、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, -C(O)R a 、-C(O)NR a R b 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b or be selected from 0-4 of the following substituents: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, -NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a Rb 、-NR a COR b or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl;

[0010] L represents a bond or C1-C6 alkylene, halogenated C1-C6 alkylene, wherein any alkylene may be replaced by -NR a 、-O-、hydroxy、halogen、-C(O)-、-C(O)O-、-OC(O)-、-NR a C(O)-、-C(O)NR a -, -S- replaced;

[0011] R L represents a ring B or ring CY-ring D group;

[0012] or -LR L represents -C1-C6 alkyl or -C1-C6 alkyl-NR a R b wherein the alkyl group is optionally replaced by one or more C1-C6 alkyl, C2-C6 alkenyl, -NR a 、-O-、Hydroxy、-C(O)-、-C(O)O-、-OC(O)-、-NR a C(O)-、-C(O)NR a -, replaced by S;

[0013] Ring B represents a C3-C6 cycloalkyl, a 3-6 membered heterocyclyl or a 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2, or 3 members selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent;

[0014] Ring C represents C3-C6 cycloalkyl, 3-6 membered heterocyclyl or 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2 or 3 members selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, C1-C6 alkoxy, -COR a, substituted by a C1-C6 alkylamino substituent;

[0015] Ring D represents a C3-C6 cycloalkyl, a 3-6 membered heterocyclyl or a 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2, or 3 members selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent;

[0016] Wherein, Y represents absence or C1-C6 alkylene; Wherein, R 3 represents hydrogen or C1-C6 alkyl; wherein m and n each independently represent an integer of 0, 1, 2, 3, or 4;

[0017] Wherein, Cy represents a 9-10 membered saturated or unsaturated fused ring; and, further, the Cy may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; and, further, the Cy may be further selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0018] Among them, R a 、R b Each independently represents hydrogen, C1-C6 alkyl,

[0019] In a preferred technical solution of the present invention, wherein the Cy represents the following group:

[0020] Wherein, in Formula I-1, Formula I-2 and Formula I-3:

[0021] Ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogenated, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-P(O)R a R b 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0022] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; X4 means CR X4 or N; X5 means CR X5 or N;

[0023] Among them, R X1 、R X2 、R X3 、R X4 、R X5 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NRa COR b or -CONR a R b .

[0024] In a preferred technical solution of the present invention, wherein the Cy represents the following group:

[0025] Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0026] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N;

[0027] Among them, R X1 、R X2 、R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a、-OC(O)NR a R b 、-NR a COR b or -CONR a R b ;

[0028] Wherein, X4 represents CH or N; X5 represents CH or N.

[0029] In a preferred technical solution of the present invention, wherein the Cy represents the following group:

[0030] Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0031] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N;

[0032] Among them, R X1 、R X2 、R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3Ra 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b ;

[0033] wherein X4 and X5 each independently represent CH or N.

[0034] In a preferred embodiment of the present invention, Cy represents the following group:

[0035] Furthermore, the Cy can be arbitrarily replaced by 0, 1, 2, or 3 deuterated, halogenated, C1-C6 alkylated, C2-C6 alkenylated, C2-C6 alkynylated, C3-C6 cycloalkylated, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0036] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N;

[0037] Among them, R X1 、R X2 、R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b, CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b .

[0038] In a preferred technical solution of the present invention, X1 represents CH or N; preferably CH.

[0039] In a preferred technical solution of the present invention, X3 represents CH or N; preferably N.

[0040] In the preferred technical solution of the present invention, wherein X2 represents CR X2 or N; where R X2 represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), -NR a R b , CN, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b .

[0041] In a preferred embodiment of the present invention, Cy represents the following group:

[0042] In a preferred technical solution of the present invention, wherein the Cy represents the following group:

[0043] Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0044] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N;

[0045] Among them, R X1 、R X2 、R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b ;

[0046] wherein X4 and X5 each independently represent CH or N.

[0047] In a preferred technical solution of the present invention, wherein the Cy represents the following group:

[0048] Furthermore, the Cy can be arbitrarily replaced by 0, 1, 2, or 3 deuterated, halogenated, C1-C6 alkylated, C2-C6 alkenylated, C2-C6 alkynylated, C3-C6 cycloalkylated, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b substituted by a substituent;

[0049] Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N;

[0050] Among them, R X1 、R X2 、R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b .

[0051] In a preferred technical solution of the present invention, X1 represents CH or N; preferably CH.

[0052] In a preferred technical solution of the present invention, X3 represents CH or N; preferably N.

[0053] In the preferred technical solution of the present invention, wherein X2 represents CR X2 or N; where R X2 represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b or -CONR a R b .

[0054] In a preferred embodiment of the present invention, Cy represents the following group:

[0055] In the preferred technical solution of the present invention, L represents -C1-C6 alkylene or -(C1-C6 alkylene)C(O)-, -C(O)(C1-C6 alkylene)-, -(C1-C6 alkylene)-(5-6 membered heteroaryl).

[0056] In a preferred technical solution of the present invention, L represents -CH2- or -CH2CH2-.

[0057] In a preferred technical solution of the present invention, L represents -CH2C(O)- or -C(O)CH2-.

[0058] In a preferred technical solution of the present invention, L represents -CH2-(5-6 membered heteroaryl)-.

[0059] In a preferred technical solution of the present invention, the 5-6 membered heteroaryl group is selected from any one of the following groups:

[0060] In the preferred technical solution of the present invention, R Lrepresents ring B, wherein ring B represents any of the following groups:

[0061] Wherein, the ring B is optionally substituted by 0, 1, 2, or 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent;

[0062] The wavy line indicates the site where L is connected.

[0063] In the preferred technical solution of the present invention, R L represents ring B, wherein ring B represents any of the following groups:

[0064] The wavy line indicates the site where L is connected.

[0065] In the preferred technical solution of the present invention, R L represents ring CY-ring D, wherein ring C represents The wavy line indicates the site connected to L, and * indicates the site connected to Y when Y is present, and * indicates the site connected to ring D when Y is absent.

[0066] In a preferred technical solution of the present invention, Y represents absence or -CH2-.

[0067] In the preferred technical solution of the present invention, wherein ring D represents Wherein, when Y is present, * represents the site of connection with Y, when Y is absent, * represents the site of connection with ring C, wherein the ring D is optionally substituted by 0, 1, 2, or 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent.

[0068] In the preferred technical solution of the present invention, wherein ring D represents

[0069] In the preferred technical solution of the present invention, R 1 express

[0070] Specifically, the present invention provides a compound having the following structure: DETAILED DESCRIPTION

[0071] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0072] INT-1: Synthesis of tert-butyl (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylate

[0073] Step 1: Dissolve (2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-5-oxopyrrolidine-3-carboxylic acid methyl ester (6 g, 14.09 mmol, synthesized using the same method as patent WO2023067355) in tetrahydrofuran (10 ml) and methanol (1 ml). The reaction mixture was cooled to 0°C and sodium borohydride (1 g, 28.18 mmol) was added. The mixture was stirred at 0°C for 1 hour. After the reaction was completed, the reaction solution was allowed to warm to room temperature, quenched by adding 2 ml of water, and extracted three times with 10 ml of ethyl acetate each time. The combined organic phases were washed with saturated brine (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo and then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 50:50) to afford tert-butyl (2-chloro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (4 g, 14.09 mmol, yield: 71.36%) as a yellow solid. LCMS (ESI): 397.86 [M+H] + , 1 H NMR(400MHz,DMSO-d6)δ8.81-8.52(m,1H),7.62–7.60(m,1H),7.41-7.37(m,2H),6.89-6.80(m,1H),4.92-4.90(m,1H),4 .67-4.60(m,1H),3.68-3.61(m,1H),3.20-3.18(m,1H),3.04(s,3H),2.36-2.32(m,2H),1.83-1.82(m,1H),1.42(s,9H).

[0074] Step 2: Dissolve tert-butyl (2-chloro-6-((2S,3S)-3-(hydroxymethyl)-N-methyl-5-oxopyrrolidine-2-carboxamido)phenyl)carbamate (4 g, 10.05 mmol) in dichloromethane (40 ml) and add triethylamine (3 g, 30.16 mmol). Cool the reaction mixture to 0°C and add methanesulfonyl chloride (2 g, 15.08 mmol). Stir the mixture at 0°C for 30 minutes. After the reaction is complete, extract with ethyl acetate three times, 10 ml each time. The combined organic phases were washed with saturated brine (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo to yield methyl ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-5-oxopyrrolidin-3-yl)methanesulfonate (4 g, 10.05 mmol, 83.59% yield) as a yellow solid. LCMS (ESI): 475.94 [M+H] +

[0075] Step 3: Methyl ((2S,3S)-2-((2-((tert-butoxycarbonyl)amino)-3-chlorophenyl)(methyl)carbamoyl)-5-oxopyrrolidin-3-yl)methanesulfonate (4 g, 8.40 mmol) was dissolved in N-methylpyrrolidone (40 ml) and anhydrous potassium carbonate (3 g, 25.21 mmol) was added. The mixture was stirred at 60°C for 12 hours. After the reaction was completed, the mixture was extracted with ethyl acetate three times with 10 ml each time. The organic phases were combined, washed with saturated brine (10 ml x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo. The product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=60:40) to give (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-carboxylic acid tert-butyl ester (2 g, 8.40 mmol, yield: 72.05%) as a yellow solid.

[0076] LCMS (ESI): 379.84 [M+H] +

[0077] 1 H NMR(400MHz,DMSO-d6)δppm 7.69–7.68(m,1H),7.64-7.62(m,1H),7.51-7.50(m,2H),4.26-4.24(m ,1H),3.95-3.92(m,1H),3.23(s,3H),3.01-2.87(m,2H),2.20-1.99(m, 2H),1.24(s,9H).

[0078] Example 1: Synthesis of (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0079] Step 1: (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylic acid tert-butyl ester INT-1 (90 mg, 0.24 mmol) was dissolved in 1,4-dioxane (2 ml) and 4-(trifluoromethyl)-6,7-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylic acid tert-butyl ester INT-1 (90 mg, 0.24 mmol) was added to 1,4-dioxane (2 ml). Hydrogen-5H-cyclopenta[b]pyridin-2-yl trifluoromethanesulfonate (103 mg, 0.31 mmol) (synthesis method reference patent WO2023125918A1), tris(dibenzylideneacetone)dipalladium (22 mg, 0.02 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (27 mg, 0.05 mmol), potassium carbonate (98 mg, 0.71 mmol). The mixture was stirred at 100°C for 2 hours. After the reaction, the mixture was extracted with ethyl acetate three times, 10 ml each time. The organic phases were combined, washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and the combined organic phases were concentrated in vacuo. The product was then purified by reverse phase column chromatography (alkaline water (0.1% ammonium bicarbonate): acetonitrile = 50%:50%) to give a yellow solid (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-carboxylic acid tert-butyl ester (100 mg, 0.24 mmol, yield: 72.39%).

[0080] LCMS (ESI): 583.02 [M+H] +

[0081] Step 2: Dissolve tert-butyl (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylate (100 mg, 0.18 mmol) in a mixture of dichloromethane (1 ml) and trifluoroacetic acid (8 ml). Stir the mixture at room temperature for 1 hour. After the reaction is complete, concentrate the reaction mixture in vacuo to remove the trifluoroacetic acid. Dissolve the mixture in ethyl acetate and adjust the pH to 8 with sodium bicarbonate. Extract with ethyl acetate three times (10 ml each time), dry over anhydrous sodium sulfate, and combine the organic phases and concentrate in vacuo. A yellow solid (3aR, 11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione was obtained (80 mg, 0.18 mmol, yield: 97.23%).

[0082] LCMS (ESI): 464.87 [M+H] +

[0083] Step 3: Dissolve (3aR,11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (80 mg, 0.17 mmol) in N,N-dimethylformamide (2 mL). Add sodium carbonate (91 mg, 0.86 mmol), tetrabutylammonium bromide (11 mg, 0.03 mmol), and 3-bromoprop-1-ene (1 g, 8.68 mmol). Stir the mixture at 100°C for 12 hours. After completion of the reaction, extract with ethyl acetate (3 times, 10 mL each). The combined organic phases were washed with saturated brine (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo and then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 30:70) to afford a yellow solid (70 mg, 0.17 mmol, 80.56%) of (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione.

[0084] LCMS (ESI): 504.93 [M+H] +

[0085] Step 4: (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (70 mg, 0.14 mmol) was dissolved in tetrahydrofuran (2 mL) and water (1 mL). Sodium periodate (890 mg, 4.16 mmol) and potassium osmate (77 mg, 0.21 mmol) were added. The mixture was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was quenched with sodium thiosulfate and extracted with ethyl acetate three times with 10 mL each. The organic phases were combined, washed with saturated sodium bicarbonate solution (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo to afford 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)acetaldehyde (60 mg, 0.14 mmol, yield: 85.38%) as a brown solid.

[0086] LCMS (ESI): 506.91 [M+H] +

[0087] Step 5: 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)acetaldehyde (60 mg, 0.12 mmol) was dissolved in methanol (2 ml), and molecular sieves (10 mg) and acetic acid (71 mg, 1.18 mmol) were added. The mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (30 mg, 0.47 mmol) was then added and reacted at room temperature for 30 minutes. After completion of the reaction, 1 ml of water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, 10 ml each time. The combined organic phases were washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by HPLC: column specifications: Kinetex 5m EVO C18, 30 mm x 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (3aR,11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (3.67 mg, 0.12 mmol, 5.17% yield) was obtained as a white solid.

[0088] LCMS (ESI): 590.90 [M+H] +

[0089] 1 H NMR(400MHz,DMSO-d6)δppm 8.12(s,1H),7.51-7.49(m,1H),7.46–7.42(m,1H),7.37(t,J=8.0Hz,1 H),4.57(d,J=9.6Hz,1H),3.49-3.47(m,1H),3.28(s,2H),3.25(s,3H), 3.04–2.95(m,5H),2.88(s,1H),2.77(s,2H),2.60–2.51(m,2H),2.41(d ,J=11.6Hz,2H),2.34(d,J=9.7Hz,2H),2.29–2.19(m,5H),2.10(s,4H).

[0090] Example 2: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0091] (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (70 mg, 0.12 mmol) was dissolved in toluene (2 ml), and methylboronic acid (213 mg, 3.55 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (19 mg, 0.02 mmol), and cesium carbonate (116 mg, 0.36 mmol) were added. The mixture was stirred at 100°C for 2 hours. Extraction was performed with ethyl acetate three times, each time with 10 ml. The organic phases were combined, washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography: column specifications: Kinetex 5m EVO C18, 30mm*150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B in 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (3aR,11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)-6,7-dihydro-5H-cyclopenta[b]pyridin-2-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (29.1 mg, 0.12 mmol, 43.06% yield) was obtained as a white solid.

[0092] LCMS (ESI): 571.35 [M+H] +

[0093] 1H NMR(400MHz,DMSO-d6)δppm 8.11(s,1H),7.23(s,3H),4.61(d,J=9.0Hz,1H),3.55–3.45(m,1H),3.23(s,3H),2.98-2.97(m,4H),2.8 6-2.84(m,4H),2.60–2.52(m,2H),2.38(d,J=11.0Hz,1H),2.34(s,3H),2.31–2.17(m,8H),2.11(s,6H).

[0094] Example 3: Synthesis of (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0095] Step 1: (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylic acid tert-butyl ester INT-1 (245 mg, 1.03 mmol) was dissolved in 1,4-dioxane (4 ml) and 5-chloro-7-( (trifluoromethyl)thiazolo[5,4-b]pyridine (300 mg, 0.79 mmol) (synthesis method reference patent WO2023125918A1), tris(dibenzylideneacetone)dipalladium (72 mg, 0.08 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (91 mg, 0.16 mmol), potassium carbonate (328 mg, 2.37 mmol). The mixture was stirred at 100°C for 2 hours. After the reaction was completed, it was extracted with ethyl acetate three times, each time with 10 ml. The organic phases were combined, washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and the combined organic phases were concentrated in vacuo. The product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate=40:60) to give a yellow solid (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-carboxylic acid tert-butyl ester (350 mg, 0.79 mmol, yield: 76.14%).

[0096] LCMS(ESI):582.00[M+H] +

[0097] Step 2: Dissolve tert-butyl (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylate (320 mg, 0.55 mmol) in a mixture of dichloromethane (1 ml) and trifluoroacetic acid (8 ml). Stir the mixture at room temperature for 1 hour. After the reaction is complete, concentrate the reaction mixture in vacuo to remove the trifluoroacetic acid. Dissolve the mixture in ethyl acetate and adjust the pH to 8 with sodium bicarbonate. Extract with ethyl acetate three times (10 ml each time), dry over anhydrous sodium sulfate, and combine the organic phases and concentrate in vacuo. A yellow solid (3aR, 11aS)-6-chloro-10-methyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione was obtained (250 mg, 0.55 mmol, yield: 94.36%).

[0098] LCMS (ESI): 481.88 [M+H] +

[0099] Step 3: Dissolve (3aR,11aS)-6-chloro-10-methyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (250 mg, 0.52 mmol) in N,N-dimethylformamide (4 mL). Add sodium carbonate (275 mg, 2.59 mmol), tetrabutylammonium bromide (33 mg, 0.10 mmol), and 3-bromoprop-1-ene (314 mg, 2.59 mmol). Stir the mixture at 100°C for 12 hours. After completion of the reaction, extract with ethyl acetate three times (10 mL each time). The combined organic phases were washed with saturated brine (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo and then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 80:20) to afford a yellow solid (220 mg, 0.52 mmol, 81.25%) of (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione).

[0100] LCMS (ESI): 521.94 [M+H] +

[0101] Step 4: (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (220 mg, 0.42 mmol) was dissolved in a mixture of tetrahydrofuran (2 mL) and water (1 mL). Sodium periodate (1353 mg, 6.32 mmol) and potassium osmate (233 mg, 0.63 mmol) were added. The mixture was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was quenched with sodium thiosulfate and extracted with ethyl acetate three times with 10 mL each. The combined organic phases were washed with saturated sodium bicarbonate solution (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo to afford 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetaldehyde as a brown solid (200 mg, 0.42 mmol, yield: 90.57%).

[0102] LCMS(ESI):523.92[M+H] +

[0103] Step 5: 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetaldehyde (80 mg, 0.15 mmol) was dissolved in methanol (2 ml), and molecular sieves (10 mg) and acetic acid (92 mg, 1.53 mmol) were added. The mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (38 mg, 0.61 mmol) was then added and reacted at room temperature for 30 minutes. After the reaction was completed, 1 ml of water was added to quench the reaction, and the mixture was extracted with ethyl acetate three times, 10 ml each time. The combined organic phases were washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by HPLC: column specifications: Kinetex 5m EVO C18, 30 mm x 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B over 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (3aR,11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (27.37 mg, 0.15 mmol, 29.07% yield) was obtained as a white solid.

[0104] LCMS (ESI): 608.05 [M+H] +

[0105] 1 H NMR(400MHz,DMSO-d6)δppm 9.63(s,1H),8.62(s,1H),7.54-7.53(m,1H),7.49-7.41(m,2H),4.69(d,J=9.5Hz,1H),3.52-3.50(m,1H),3.27(s,3H),3. 02-3.01(m,3H),2.83(d,J=11.8Hz,1H),2.61(t,J=8.4Hz,1H),2.45(s,1H),2.41(s,1H),2.34–2.18(m,9H),2.10(s,3H).

[0106] Example 4: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0107] (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (80 mg, 0.13 mmol) was dissolved in toluene (2 ml), and methylboronic acid (236 mg, 3.95 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (21 mg, 0.03 mmol), and cesium carbonate (129 mg, 0.39 mmol) were added. The mixture was stirred at 100°C for 2 hours. Extraction was performed with ethyl acetate three times, each time with 10 ml. The organic phases were combined, washed with saturated brine (10 ml x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography: column specifications: Kinetex 5m EVO C18, 30mm*150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min; gradient: 20% B to 47% B in 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 9.35. (3aR,11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (6.54 mg, 0.13 mmol, 8.40% yield) was obtained as a white solid.

[0108] LCMS (ESI): 588.10 [M+H] +

[0109] 1H NMR(400MHz,DMSO-d6)δppm 9.62(s,1H),8.62(s,1H),7.32–7.23(m,3H),4.73(d,J=8.8Hz,1H),3.53(d,J=9.8Hz,1H),3.24(s,3H),2.99(d,J=6.8Hz,1H), 2.92–2.83(m,3H),2.63(d,J=9.0Hz,1H),2.46(s,1H),2.43(d,J=6.0Hz,1H),2.36(s,3H),2.26(t,J=6.8Hz,9H),2.11(s,3H).

[0110] Example 5: Synthesis of (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11aS hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0111] Step 1: (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylic acid tert-butyl ester INT-1 (0.20 g, 0.53 mmol) was dissolved in 1,4-dioxane (1.00 ml) and potassium carbonate (0.22 g, 1.59 mmol) was added. ), 4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl trifluoromethanesulfonate (0.18 g, 0.51 mmol) (synthesis method reference patent WO2023125918A1), tris(dibenzylideneacetone)dipalladium (0.05 g, 0.05 mmol) and 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (0.06 g, 0.10 mmol), after nitrogen replacement, the reaction was stirred at 100 ° C for 2 hours. After completion of the reaction, the reaction solution was poured into water (2 mL), extracted with ethyl acetate (2 mL*2), dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain the crude product, which was purified by silica gel column chromatography (petroleum ether:ethyl acetate=30:70) to obtain (3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-carboxylic acid tert-butyl ester (0.15 g, 0.26 mmol, 48% yield) as a yellow oil.

[0112] LCMS(ESI):581[M+H]+

[0113] Step 2: Dissolve (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-5-carboxylic acid tert-butyl ester (0.15 g, 0.26 mmol) in trifluoroacetic acid (1.80 ml) and add dichloromethane (0.2 ml). Stir the reaction at room temperature for 1 hour. After the reaction is complete, the reaction solution is added to saturated aqueous ammonium bicarbonate. Extract with ethyl acetate (2×5 ml). Wash the combined organic layers with saturated brine (2×5 ml) and dry over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give (3aR, 11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (0.11 g, 0.23 mmol, 92% yield) as a yellow solid.

[0114] LCMS(ESI):481[M+H]+,

[0115] 1 H NMR(400MHz,DMSO-d6)δppm 8.50(s,1H),8.06-8.04(m,1H),7.82-7.78(m,1H),7.54–7.42(m,2H),7.37-7.32(m,1H),4. 81-4.80(m,1H),4.47-4.45(m,1H),3.54-3.52(m,3H),3.23-3.20(m,2H),2.80-2.64(m,3H).

[0116] Step 3: Dissolve (3aR, 11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (0.11 g, 0.23 mmol) in N,N-dimethylformamide solution (2.00 ml), add 3-bromopropene (1.51 g, 12.58 mmol), sodium carbonate (0.08 g, 0.75 mmol) and tetrabutylammonium bromide (0.01 g, 0.03 mmol), and stir the reaction at 100°C for 2 hours. After completion of the reaction, the reaction solution was poured into water (2 mL), extracted with ethyl acetate (3 mL*3), dried over anhydrous sodium sulfate, filtered, and the organic phase was spin-dried and purified by silica gel column chromatography (petroleum ether:ethyl acetate=20:80) to give (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (0.10 g, 0.19 mmol, yield: 69%) as a yellow solid.

[0117] LCMS(ESI):521[M+H]+

[0118] Step 4: (3aR,11aS)-5-allyl-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (0.10 g, 0.19 mmol) was dissolved in tetrahydrofuran solution (2.00 mL) and water (0.4 mL). Sodium periodate (1.23 g, 5.75 mmol) and potassium osmate (0.07 g, 0.19 mmol) were added and stirred at room temperature for 1 hour. After completion of the reaction, the reaction was quenched by the addition of saturated sodium thiosulfate solution (4 mL) at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 5 mL). The combined organic layers were washed with saturated brine (2 x 5 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to provide 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetaldehyde (60 mg, 0.11 mmol, 60% yield) as a black solid.

[0119] LCMS(ESI):523[M+H]+

[0120] Step 5: 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetaldehyde (60 mg, 0.11 mmol) was dissolved in methanol (2.00 ml), 1-methylpiperazine (23 mg, 0.23 mmol) and acetic acid (69 mg, 11.49 mmol) were added and stirred at room temperature for 20 minutes. Sodium cyanoborohydride (29 mg, 0.46 mmol) was then added at 0°C, and the reaction was stirred at room temperature for 30 minutes. After the reaction was completed, the reaction solution was poured into water (5 ml), extracted with ethyl acetate (5 ml * 2), the organic phase was spin-dried, and the crude product was purified by high pressure preparative chromatography: column: Kinetex 5m EVO C18, 30 mm*150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 29% B to 49% B in 7 minutes; wavelength: 254 nm / 220 nm, retention time: 5.12 minutes; to obtain (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (7.14 mg, 0.01 mmol, 10% yield) as a white solid.

[0121] LCMS (ESI): 606.90 [M+H] +

[0122] 1H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.06(d,J=6.2Hz,1H),7.56-7.50(m,1H),7.49–7.47(m,1H ),7.46-7.41(m,2H),4.70-4.67(m,1H),3.54-3.52(m,1H),3.50-3.48(m ,1H),3.28(s,4H),3.12–2.97(m,4H),2.89-2.80(m,1H),2.62(dd,J=8.4 Hz,1H),2.47-2.39(m,3H),2.32(m,1H),2.34–2.21(m,4H),2.10(s,3H).

[0123] Example 6: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0124] (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (80 mg, 0.13 mmol) was dissolved in Methylboronic acid (237 mg, 3.95 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (21 mg, 0.01 mmol) and cesium carbonate (129 mg, 0.40 mmol) were added to toluene (2.0 ml), the atmosphere was replaced with nitrogen, and the reaction was stirred at 100°C for 2 hours. After the reaction was completed, the reaction solution was poured into water (3 ml), extracted with ethyl acetate (3 ml * 2), the organic phase was spin-dried, and the crude product was purified by high pressure preparative chromatography (column: XBridge BEH Shield RP18 5m, 30mm×150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 41% B to 71% B in 7 minutes; wavelength: 254 nm / 220 nm, retention time: 6.12 minutes; to obtain (3aR,11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (9.36 mg, 0.02 mmol, 12% yield) as a white solid.

[0125] LCMS (ESI): 587.25 [M+H] +

[0126] 1 H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05(d,J=6.0Hz,1H),7.48-7.47(m,1H),7.30–7.25(m,3H),4.73(d,J=8.8Hz,1H),3.52(d,J=9.8Hz,1H),3.25(s,3H),3.04–2 .95(m,1H),2.93–2.82(m,3H),2.68–2.59(m,1H),2.49(s,1H),2.46-2.39(dd,J=10.8Hz,2H),2.35(s,3H),2.33-2.24(m,8H),2.11(s,3H).

[0127] Example 7: Synthesis of (3aR, 11aS)-5-(2-((R)-3-hydroxypyrrolidin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0128] Referring to Example 5, (R)-pyrrolidin-3-ol was used in place of 1-methylpiperazine to give (3aR,11aS)-5-(2-((R)-3-hydroxypyrrolidin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (3.21 mg, 0.18 mmol, 3.00% yield) as a white solid.

[0129] LCMS(ESI):573.63[M+H] +

[0130] 1 H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05(d,J=6.2Hz,1H),7.48(s,1H),7.29(d,J=5.0Hz,3H),4.73(d,J=8.8Hz,1H),4 .62(d,J=4.6Hz,1H),4.12-4.11(m,2H),3.52(s,1H),3.25(s,3H),3.15(s,1H),3.01-3.00(m,1 H),2.88(d,J=13.0Hz,1H),2.82-2.81(m,1H),2.64(d,J=20.8Hz,2H),2.38-2.37(m,3H),2.34( d,J=6.6Hz,3H),2.24-2.22(m,1H),1.91(d,J=5.8Hz,1H),1.60–1.42(m,1H),1.31-1.30(m,1H).

[0131] Example 8: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-morpholinoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0132] Referring to Examples 5 and 6, morpholine was used to replace 1-methylpiperazine to give (3aR, 11aS)-6,10-dimethyl-5-(2-morpholinoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (10.42 mg, 26.93% yield) as a white solid.

[0133] LCMS(ESI):573.63[M+H] +

[0134] 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.50(d,J=2.0Hz,1H),8.05(m,1H),7.49(d,J=6.2Hz,1H),7.29(d,J=5.1Hz ,3H),4.74(d,J=8.3Hz,1H),3.52-3.51(m,5H),3.26(d,J=2.0Hz,3H),3.02 -3.01(m,1H),2.92(d,J=12.2Hz,2H),2.88-2.86(m,2H),2.67(s,1H),2.61 (d,J=6.4Hz,2H),2.44(s,1H),2.37(d,J=2.2Hz,3H),2.28(d,J=7.8Hz,3H).

[0135] Example 9: Synthesis of (3aR, 11aS)-5-(2-(4-acetylpiperazin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0136] Referring to Example 5, 1-methylpiperazine was replaced with acetylpiperazine to give (3aR, 11aS)-5-(2-(4-acetylpiperazin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (7.60 mg, 6% yield) as a white solid.

[0137] LCMS (ESI): 615.15 [M+H]+

[0138] 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.49(s,1H),8.05(d,J=6.2Hz,1H),7.48-7.46(m,1H),7.31-7.26(m,3H),4.7 3(d,J=8.8Hz,1H)3.52(m,2H),3.38-3.36(m,2H),3.26(s,3H),3.05-3.01(m,1 H),2.97-2.93(d,J=13.0Hz,1H),2.92-2.88(d,J=11.0Hz,1H),2.67-2.60(m,2 H),2.43(m,2H),2.37(s,3H),2.31(m,4H),2.25(d,J=5.8Hz,2H),1.96(s,3H).

[0139] Example 10: Synthesis of (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0140] Referring to Example 5, 1-(methylsulfonyl)piperazine was substituted for 1-methylpiperazine to give (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (8.78 mg, 3% yield) as a white solid.

[0141] LCMS (ESI): 670.90 [M+H] +

[0142] 1H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05(d,J=6.1Hz,1H),7.55(m,1H),7.49(m,1H),7.47(m,2H),4.71(d,J=9.0Hz,1H),3.57( m,2H),3.28(s,4H),3.05-3.03(m,4H),2.84(s,3H),2.67(m,2H),2.49(s,3H),2.43(m,2H),2.32(s,3H).

[0143] Example 11: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0144] With reference to Example 6, (3aR, 11aS)-6,10-dimethyl-5-(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (3.05 mg, 21% yield) was obtained as a white solid.

[0145] LCMS (ESI): 650.90 [M+H] +

[0146] 1 H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05-8.02(m,1H),7.52–7.45(m,1H),7.34–7.24(m,3H),4.74(d,J=9.0Hz,1H),3.54(m,2H),3.05(s,3H),3.04(m,4H ),3.00(m,1H),2.93(m,1H),2.89(d,J=12.0Hz,1H),2.86(s,3H),2.67(m,1H),2.62(m,1H),2.42(s,4H),2.36(s,3H),2.30(d,2H).

[0147] Example 12: Synthesis of (3aR, 11aS)-5-(2-(3,8-diazabicyclo[3.2.1]octan-3-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0148] Step 1: Referring to Example 5, tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate was substituted for 1-methylpiperazine to give tert-butyl 3-(2-((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (73 mg, 0.10 mmol, 52.52% yield) as a yellow solid.

[0149] LCMS(ESI):699[M+H] +

[0150] Step 2: Tert-butyl 3-(2-((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (73 mg, 0.10 mmol) was dissolved in trifluoroacetic acid (1.8 mL), and dichloromethane (0.2 mL) was added. The mixture was stirred at 60° C. for 20 minutes. The reaction progress was monitored by LCMS. The reaction solution was concentrated, and the crude product was purified by preparative HPLC: column: Sunfire C18 5 m, 30 mm × 150 mm; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 15% B to 40% B in 10 minutes; wavelength: 254 nm / 220 nm; RT1 (min): 9. (3aR,11aS)-5-(2-(3,8-diazabicyclo[3.2.1]octan-3-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (29.24 mg, 57% yield) was obtained as a white solid.

[0151] LCMS (ESI): 599.60 [M+H] +

[0152] 1 H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05(d,J=6.2Hz,1H),7.49-7.47(m,1H),7.33-7.26(m,3 H),4.73(d,J=8.8Hz,1H)3.62(s,2H),3.54-3.53(m,1H),3.26(s,3H),3 .01-2.95(m,1H),2.91-2.82(m,3H),2.67-2.57(m,3H),2.43(m,1H),2 .28(d,J=5.8Hz,3H),2.19(d,2H),1.89-1.88(m,2H)1.77-1.68(m,4H).

[0153] Example 13: Synthesis of (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)selenophenyl[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0154] With reference to Example 5, 6-chloro-4-(trifluoromethyl)seleno[2,3-b]pyridine-2-carboxylic acid ethyl ester (synthesis method reference patent: WO2023125918A1) was used to replace 4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yltrifluoromethanesulfonic acid to obtain compound 1, which was then subjected to reductive amination to obtain (3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)selenophenyl[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (3.34 mg, yield 5.80%) as a white solid.

[0155] LCMS(ESI):653.99[M+H] +

[0156] 1 H NMR(400MHz,DMSO-d6)δppm 8.54(d,J=6.6Hz,1H),8.47(s,1H),7.69–7.61(m,1H),7.54(dd,J=6.2,3.4Hz,1H),7 .49–7.40(m,2H),4.69(d,J=9.5Hz,1H),3.53(d,J=4.7Hz,1H),3.3(s,3H),3.06–2.9 7(m,3H),2.83(s,1H),2.60(dd,J=16.6,8.4Hz,2H),2.47(d,J=5.0Hz,1H),2.40(s,2 H),2.33(t,J=1.9Hz,2H),2.27(s,1H),2.26(s,2H),2.25–2.21(m,2H),2.11(s,3H).

[0157] Example 14: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)selenophenyl[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0158] Using a similar method to Example 6, (3aR,11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(4-(trifluoromethyl)selenophenyl[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (2.68 mg, 6.86% yield) was obtained as a white solid. LCMS (ESI): 633.57 [M+H] +

[0159] 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.49(d,J=6.4Hz,1H),8.06-8.04(s,1H),7.49-7.48(d,J=6.4Hz,1H),7.33-7 .32(m,2H),7.27-7.26(m,1H),6.93-6.89(m,1H),6.22-6.17(m,1H),5.79-5.7 6(m,1H),4.81-4.75(m,2H),4.31-4.28(m,1H),4.17-4.13(m,1H),4.12-4.09 (m,1H),3.95-3.94(m,1H),3.49-3.46(m,1H),3.31-3.29(m,3H),3.18-3.13(m 2H),2.96-2.88(m,2H),2.63-2.61(m,1H),2.49-2.48(m,2H),2.28-2.22(m,3H).

[0160] Example 15: Synthesis of (3aR, 11aS)-5-((7-acryloyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazopyrimidine-2,11(3H)-dione

[0161] Step 1: Dissolve tert-butyl 3-formyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (315 mg, 1.25 mmol) in dimethylformamide (5 mL). Add (3aR,11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (200 mg, 0.42 mmol). Cool the reaction mixture to 0°C and add trimethylsilyl chloride (113 mg, 1.04 mmol). Stir the mixture at 0°C for 30 minutes. Borane tetrahydrofuran complex (36 mg, 0.42 mmol) was added, and the mixture was stirred at 0°C for another 2 hours. After the reaction was complete, the mixture was extracted with ethyl acetate three times (10 mL each time). The combined organic phases were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, and concentrated in vacuo. The product was then purified by silica gel column chromatography (alkaline water:acetonitrile = 1:1) to give tert-butyl 3-(((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11aSoctahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (220 mg, yield 73.76%). LCMS (ESI): 717.16 [M+H]. + .

[0162] The second step: tert-butyl 3-(((3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazooctan-5-yl)methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7-yl)- 8H)-tert-Butyl carboxylate (200 mg, 0.28 mmol) was dissolved in 1,4-dioxane (3 ml), and methylboronic acid (501 mg, 8.37 mmol), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (44 mg, 0.06 mmol), and anhydrous potassium carbonate (116 mg, 0.84 mmol) were added. The mixture was stirred at 100°C for 2 hours. After completion of the reaction, the mixture was extracted three times with 10 ml of ethyl acetate each time. The combined organic phases were washed with saturated brine (10 mL x 3) and dried over anhydrous sodium sulfate. The combined organic phases were concentrated in vacuo to afford tert-butyl 3-(((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoindane-5-yl)methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (130 mg, yield: 66.90%). LCMS (ESI): 696.75 [M+H]. +

[0163] Step 3: Dissolve tert-butyl 3-(((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoindane-5-yl)methyl)-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (100 mg, 0.07 mmol) in dichloromethane (3 mL) and trifluoroacetic acid (1 mL). The mixture was stirred at 60°C for 1 hour. After completion of the reaction, the mixture was concentrated in vacuo. The product was then purified by silica gel column chromatography (alkaline water:acetonitrile = 1:1) to afford (3aR,11aS)-6,10-dimethyl-5-((5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazopyrimidine-2,11(3H)-dione (80 mg, yield: 72.39%). LCMS (ESI): 596.63 [M+H]. +

[0164] Step 4: (3aR,11aS)-6,10-dimethyl-5-((5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazopyrimidine-2,11(3H)-dione (60 mg, 0.10 mmol) was dissolved in N,N-dimethylacetamide (2 mL). Acrylic acid (7 mg, 0.10 mmol), propylphosphonic anhydride (96 mg, 0.30 mmol), and N,N-diisopropylethylamine (78 mg, 0.60 mmol) were added sequentially. The mixture was stirred at room temperature for 1 hour. After the reaction, the mixture was extracted with ethyl acetate three times (10 mL each time). The combined organic phases were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by HPLC: column specifications: Kinetex 5m EVO C18, 30 mm x 150 mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 34% B to 50% B over 10 minutes; wavelength: 254 / 220 nm; retention time (minimum): 7.35. (3aR,11aS)-5-((7-acryloyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazopyrimidine-2,11(3H)-dione (10.04 mg, 14.83% yield) was obtained as a white solid. LCMS (ESI): 650.68 [M+H] +

[0165] 1H NMR(400MHz,DMSO-d6)δppm 8.50(s,1H),8.05(d,J=6.1Hz,1H),7.52-7.45(m,1H),7.33(d,J=5.8Hz,2H),7.27(d,J=5.4H z,1H),6.92(s,1H),6.22(s,1H),5.77(s,1H),4.76(d,J=9.0Hz,2H),4.28(s,1H),4.15(d,J= 14.6Hz,1H),4.09(s,1H),3.95(s,1H),3.48(d,J=10.4Hz,1H),3.27(s,3H),3.24-3.22(m,2H ),2.98(s,1H),2.91(d,J=6.4Hz,1H),2.60(d,J=9.8Hz,1H),2.44-2.43(m,2H),2.29(s,3H).

[0166] Example 16: Synthesis of (3aR, 11aS)-5-(2-(4-(1-acryloylazine-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0167] The first step: 2-{(11aS, 3aR)-6,10-dimethyl-2,11-dioxo-1-[7-(trifluoromethyl)(1,3-thiazolo[4,5-e]pyridin-5-yl)]-4H, 11aH, 3aH-benzo[b]pyrrolidinone[2,3-f]1,4-diazooctan-5-yl}acetaldehyde (90 mg, 0.18 mmol) was dissolved in dichloromethane (10.00 ml) and 3-(piperazine-1-yl)azetidine-1-carboxylic acid benzyl ester (656 mg, 2.39 mmol) was added to the solution and stirred at room temperature for 2 hours. Then, sodium cyanoborohydride (188 mg, 2.98 mmol) was added at 0°C and stirred at room temperature overnight. The reaction progress was monitored by LCMS. After completion of the reaction, the reaction was quenched by adding aqueous ammonium bicarbonate (20 ml) at room temperature. The resulting mixture was extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with saturated brine (2 x 20 ml), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase flash chromatography (column, C18 silica gel; mobile phase, acetonitrile: aqueous ammonium bicarbonate solution (0.01%) = 20%:80%) to give benzyl 3-(4-(2-((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)ethyl)piperazin-1-yl)azetidine-1-carboxylate (100 mg, 0.13 mmol, 66.23% yield) as a yellow solid.

[0168] LCMS(ESI):763[M+H]+

[0169] Step 2: Benzyl 3-(4-(2-((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)ethyl)piperazin-1-yl)azetidine-1-carboxylate (100 mg, 0.13 mmol) was dissolved in hydrochloric acid (2.00 mL) and stirred at 80° C. for 1 hour. The reaction progress was monitored by LCMS. The reaction solution was concentrated under reduced pressure to give (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (80 mg, 0.13 mmol, 97% yield) as a black solid.

[0170] LCMS(ESI):629[M+H]+

[0171] Step 3: (3aR,11aS)-5-(2-(4-(azetidin-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (80 mg, 0.13 mmol) was dissolved in N,N-dimethylacetamide (2.00 mL). Acrylic acid (10 mg, 0.14 mmol), 1-n-propylphosphoric anhydride (94 mg, 0.30 mmol), and N,N-diisopropylethylamine (111 mg, 0.86 mmol) were added and stirred at room temperature for 1 hour. The reaction progress was monitored by LCMS. After the reaction was completed, the reaction mixture was poured into water (2 ml), extracted with ethyl acetate (2 ml * 2), and the reaction solution was concentrated. The crude product was purified by high performance liquid chromatography: column: YMC Triart C18 ExRs 5m, 30mm×150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 37% B to 45% B in 10 minutes; wavelength: 254 nm / 220 nm; to obtain (3aR,11aS)-5-(2-(4-(1-acryloylazine-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(7-(trifluoromethyl)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro 2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (10.44 mg, 10.50% yield) as a white solid.

[0172] LCMS (ESI): 683.00 [M+H]+

[0173] 1H NMR(400MHz,DMSO-d6)δppm 9.62(s,1H),8.62(s,1H),7.34-7.21(m,3H),6.31-6.25(m,1H),6.08-6.06(m,1H),5.66-5.63(m,1H),4.73(d,J =8.8Hz,1H),4.19(t,J=8.0Hz,1H),3.98-3.97(m,1H),3.91-3.85(m,1H),3.70-3.66(m,1H),3.53(d,J=9.8Hz,1 H),3.24(s,3H),3.09-3.06(m,1H),3.02-2.97(m,1H),2.95-2.87(m,1H),2.85-2.83(m,2H),2.67-2.58(m,1H), 2.44(s,2H),2.42(m,1H),2.36(s,3H),2.32(m,2H),2.29(t,J=6.8Hz,1H),2.27-2.26(m,4H),2.25-2.24(m,1H).

[0174] Example 17: Synthesis of (3aR, 11aS)-5-(2-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0175] A similar method to that of Reference Example 16 was used to obtain (3aR, 11aS)-5-(2-(4-(1-acryloylazetidin-3-yl)piperazin-1-yl)ethyl)-6,10-dimethyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (6.81 mg, 28% yield) as a white solid.

[0176] LCMS (ESI): 682.00 [M+H]+

[0177] 1H NMR(400MHz,DMSO-d6)δppm 8.49(s,1H),8.05(d,J=6.0Hz,1H),7.56-7.42(m,1H),7.35-7.23(m,3H),6.31-6.24(m,1H),6.08(dd,J=17.0,2. 3Hz,1H),5.65(dd,J=10.3,2.2Hz,1H),4.74-4.68(m,1H),4.19(t,J=8.0Hz,1H),4.02-3.94(m,1H),3.94-3.85(m, 1H),3.68(dd,J=10.4,5.1Hz,1H),3.52(d,J=9.9Hz,1H),3.27(s,1H),3.24(s,3H),3.11-2.94(m,3H),2.92-2.90( m,1H),2.66-2.58(m,2H),2.43(s,2H),2.41(m,1H),2.38(m,2H),2.36(s,3H),2.28(t,J=6.9Hz,4H),2.26(m,1H).

[0178] Example 18: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0179] Step 1: ((3aR,11aS)-6-chloro-10-methyl-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (200 mg, 0.42 mmol) was dissolved in N-methylpyrrolidone (5.00 ml) and butyl bromoacetate (812 mg, 4.16 mmol), potassium carbonate (172 mg, 1.25 mmol) and tetrabutylammonium iodide (15.4 mg, 0.04 mmol) were added. The mixture was stirred at 120°C overnight. The reaction progress was monitored by LCMS. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was dried by rotary evaporation and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 70%:30%) to afford 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetic acid (80 mg, 0.15 mmol, 36% yield) as a yellow solid.

[0180] LCMS(ESI):539[M+H]+

[0181] Step 2: 2-((3aR,11aS)-6-chloro-10-methyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)acetic acid (80 mg, 0.15 mmol) was dissolved in dichloromethane (2.00 mL). 1-Methylpiperazine (30 mg, 0.30 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65 mg, 0.17 mmol), and N,N-diisopropylethylamine (55 mg, 0.43 mmol) were added to the solution and stirred at room temperature for 1 hour. The reaction progress was monitored by LCMS. After the reaction was complete, water (5 mL) was added to terminate the reaction at room temperature. The resulting mixture was extracted with dichloromethane (2 × 5 mL). The combined organic layers were washed with saturated brine (2 × 5 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 50%:50%) to afford (3aR,11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (90 mg, 0.15 mmol, 97% yield) as a white oil.

[0182] LCMS(ESI):621[M+H]+

[0183] Step 3: ((3aR, 11aS)-6-chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (90 mg, 0.15 mmol) was dissolved in 1,4-dioxane (2.00 ml) and added Methylboronic acid (261 mg, 4.35 mmol), potassium carbonate (60 mg, 0.43 mmol) and chloro[(4,5-bis(diphenylphosphino)-9,9-dimethylxanthene)-2-(2-aminobiphenyl)]palladium(II) (23 mg, 0.03 mmol) were replaced with nitrogen and stirred at 100°C for 1 hour. The reaction progress was monitored by LCMS. After completion of the reaction, the reaction solution was poured into water and extracted with ethyl acetate. The organic phase was dried by spin drying and the crude product was purified by HPLC: Column: XBridge BEH Shield RP18 5m, 30mm×150mm; mobile phase A: water (10 mmol / L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml / min ml / min; gradient: from 40% B to 56% B in 8 minutes; wavelength: 254 nm / 220 nm; (3aR,11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)-2-oxoethyl)-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (18.08 mg, 21% yield) was obtained as a white solid.

[0184] LCMS (ESI): 601.00 [M+H]+

[0185] 1H NMR(400MHz,DMSO-d6)δppm 8.50(s,1H),8.05(d,J=6.2Hz,1H),7.48-7.47(m,1H),7.35-7.23(m,3H),4.76(d,J =8.9Hz,1H),3.79(d,J=15.0Hz,1H),3.64(d,J=15.0Hz,1H),3.51(d,J=10.2Hz,1H) ,3.46-3.45(m,2H),3.20(s,3H),3.01-2.91(m,2H),2.67-2.66(m,1H),2.62-2.61( m,2H),2.50-2.49(m,2H),2.42(s,3H),2.26-2.25(m,2H),2.22(s,1H),2.16(s,3H).

[0186] Example 19: Synthesis of (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)furo[3,2-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione

[0187] A similar method to that of Reference Example 6 was used to obtain (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)furo[3,2-b]pyridin-5-yl)-1,3a,4,5,10,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazooctane-2,11(3H)-dione (5.51 mg, 28.52% yield) as a white solid.

[0188] LCMS (ESI): 571.15 [M+H] +

[0189] 1H NMR(400MHz,DMSO-d6)δppm 8.52(d,J=2.4Hz,1H),8.41(s,1H),7.54-7.47(m,1H),7.45(s,1H),7.33-7.22(m,2H),4.69(d,J=8.8Hz,1H),3.51(d,J=10.2Hz,2H),3.30( s,3H),3.16(m,2H),2.99(m,2H),2.93-2.82(m,2H),2.35(m,1H),2.30 (s,3H),2.27(d,J=7.0Hz,2H),2.23(d,4H),2.20(m,2H),2.14(s,3H).

[0190] Example 20: Synthesis of tert-butyl 4-(4-(((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)methyl)oxazol-2-yl)piperazine-1-carboxylate

[0191] A similar method as in Example 15 was used to obtain tert-butyl 4-(4-(((3aR,11aS)-6,10-dimethyl-2,11-dioxo-1-(4-(trifluoromethyl)thieno[2,3-b]pyridin-6-yl)-1,2,3,3a,4,10,11,11a-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazoxazin-5-yl)methyl)oxazol-2-yl)piperazine-1-carboxylate (1.05 mg, 5%).

[0192] LCMS (ESI): 725.95 [M+H] +

[0193] 1 H NMR(400MHz,DMSO-d6)δppm 8.49-8.47(m,1H),8.08-8.03(m,1H),7.57-7.45(m,2H),7.42-7.35(m,2H),7.25- 7.20(m,1H),4.93-4.83(m,1H),3.75-3.65(m,1H),3.35-3.32(m,4H),3.28-3.27(m ,3H),3.22-3.20(m,2H),3.19-2.93(m,2H),2.92-2.90(m,2H),2.50-2.49(m,3H), 2.35-2.34(m,1H),1.95-1.88(m,1H),1.70(s,1H),1.41-1.40(m,9H),1.23(s,1H).

[0194] Example 21: (3aR, 11aS)-6-Chloro-10-methyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thio)thiazol[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione.

[0195] Step 1: Dissolve 4-bromo-2,6-dichloro-3-pyridinamine (20.0 g, 82.68 mmol) in 1,4-dioxane (100.0 mL), add oxyphenylmethane isothiocyanate (20.24 g, 124.02 mmol), and stir at 100°C overnight. LCMS showed the product as the main product. The reaction was cooled to room temperature and concentrated. The residue was recrystallized from dichloromethane and filtered to give N-(7-bromo-5-chloro(1,3-thiazolo[5,4-b]pyridin-2-yl))benzamide (30 g, 98%) as an off-white solid.

[0196] LCMS(ESI):[M+H] + =367.74 / 369.65.

[0197] Step 2: N-(7-bromo-5-chloro(1,3-thiazolo[5,4-b]pyridin-2-yl))benzamide (30.5 g, 82.74 mmol) was added to a solution of concentrated sulfuric acid (70%) (300.00 mL). Stirred at 140 ° C for 15 minutes. LCMS showed 50% product and complete consumption of the starting material. The mixture was poured into water and the pH was adjusted to 8-9 with potassium hydroxide (1.0 N aqueous solution). A white precipitate was precipitated, filtered, and the filter cake was washed with water to collect the crude product. The crude product was dissolved in tetrahydrofuran and filtered. The filtrate was concentrated to give 7-bromo-5-chloro-1,3-thiazolo[5,4-b]pyridin-2-amine (18.90 g, 43%) as an off-white solid.

[0198] LCMS(ESI):[M+H] + =263.90 / 265.87

[0199] Step 3: 7-bromo-5-chloro-1,3-thiazolo[5,4-b]pyridin-2-amine (20.5 g, 38.75 mmol) was dissolved in tetrahydrofuran (200.00 mL), tert-butyl nitrite (11.3 g, 107.74 mmol) was added, and the mixture was stirred at 70°C overnight. LCMS showed that the product was the main product and the starting material was completely consumed. The mixture was poured into water (400 mL) and extracted with ethyl acetate (200 mL × 3). The organic layers were combined, dried, and concentrated. The residue was purified by silica gel column chromatography (eluted with 5% to 20% ethyl acetate / petroleum ether solution) to obtain 7-bromo-5-chloro-1,3-thiazolo[5,4-b]pyridine (8.00 g, 74%) as a yellow solid.

[0200] LCMS(ESI):[M+H] + =248.8 / 250.8

[0201] Step 4: Dissolve 7-bromo-5-chloro-1,3-thiazolo[5,4-b]pyridine (500 mg, 2.00 mmol) in acetonitrile (10.0 mL), add trimethylsilyl chloride (261 mg, 2.40 mmol) and sodium iodide (350 mg, 2.40 mmol), respectively. The reaction mixture was stirred at 80°C for 3 hours. LCMS showed 30% starting material remaining and approximately 60% product. The reaction was cooled to room temperature and concentrated. The residue was purified by flash column chromatography (silica gel, 0-5% gradient of petroleum ether / ethyl acetate) to afford 5-chloro-7-iodo-1,3-thiazolo[5,4-b]pyridine (310 mg, 52%) as a yellow solid.

[0202] LCMS(ESI):[M+H] + =296.8.

[0203] Step 5: Dissolve 5-chloro-7-iodo-1,3-thiazolino[5,4-b]pyridine (320 mg, 1,08 mmol) in 1,4-dioxane (10,0 mL). Add trifluoromethylthio(2,2-bipyridine)copper(I) (451 mg, 1,40 mmol) and stir the reaction mixture at 110°C for 16 hours. Monitor the reaction by TLC (petroleum ether / ethyl acetate = 10 / 1). TLC shows a small amount of raw material remaining and 70% of the product (new point with less polarity). The reactants were filtered, concentrated and purified by flash column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give 5-chloro-7-(trifluoromethylthio)-1,3-thiazolino[5,4-b]pyridine (210 mg, 50%) as a yellow solid. By 1 HNMR and 19 FNMR confirmed the structure.

[0204] LCMS(ESI):[M+H] + =271.0.

[0205] 1 H NMR (400MHz, DMSO-d6) δppm 9.71 (s, 1H), 7.91 (s, 1H).

[0206] 19 FNMR (400MHz, DMSO-d6) δppm-39.06 (s, 3F).

[0207] Step 6: Methyl (2S,3S)-2-(N-{2-[(tert-butoxy)carbonylamino]-3-chlorophenyl}-N-methylcarbamoyl)-5-oxopyrrolidine-3-carboxylate (5.00 g, 11.74 mmol) was dissolved in tetrahydrofuran (5.00 mL) and ethanol (5.00 mL). Solid sodium borohydride (0.96 g, 23.48 mmol) was slowly added portionwise at -5°C. TLC (dichloromethane / methanol = 20:1) monitored the reaction over 30 minutes, indicating 20% ​​starting material remaining. The reaction was quenched by slowly adding aqueous ammonium chloride dropwise with stirring for 30 minutes, controlling the temperature between -5°C and 0°C. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with saturated brine (20 mL), and the organic phase was dried over anhydrous sodium sulfate. After concentration, column chromatography (dichloromethane / methanol = 20:1) gave a white solid [(2S,3S)-3-(hydroxymethyl)-5-oxopyrrolidin-2-yl]-N-{2-[(tert-butoxy)carbonylamino]-3-chlorophenyl}-N-methylformamide (3.10 g, yield: 66%).

[0208] LCMS(ESI):[M+H] + =398.1.and[2M+H] + =795.5.

[0209] 1 H NMR(400MHz,DMSO-d6)δppm 8.75(s,1H),7.61-7.38(m,3H),4.65-4.64(m,1H),4.08-4.05(m,1H),3.65-3.64(m,1 H),3.17(s,3H),3.07-3.04(m,2H),2.50-2.43(m,2H),1.86-1.82(m,1H),1.42(s,9H).

[0210] Step 7: [(2S,3S)-3-(hydroxymethyl)-5-oxopyrrolidin-2-yl]-N-{2-[(tert-butoxy)carbonylamino]-3-chlorophenyl}-N-methylformamide (10.62 g, 26.64 mmol) was dissolved in dichloromethane (200 mL) and triethylamine (17.8 g, 175.95 mmol) was added. Methanesulfonyl chloride (6.05 g, 52.78 mmol) was then added dropwise at 0°C. The reaction mixture was stirred at 0°C for 1 hour. The reaction was monitored by LCMS and TLC (dichloromethane / methanol = 20:1). After completion of the reaction, water (100 mL) was added to quench the reaction and the mixture was extracted with ethyl acetate (2 x 200 mL). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated to give methyl [(2S,3S)-2-(N-{2-[(tert-butoxy)carbonylamino]-3-chlorophenyl}-N-methylcarbamoyl)-5-oxopyrrolidin-3-yl]methanesulfonate (11.40 g, 76%) as a white solid, which was used directly in the next step without further purification.

[0211] LCMS(ESI):[M-100+H] + and[M-55+H] + =376.2and420.2.

[0212] Step 8: Methyl [(2S,3S)-2-(N-{2-[(tert-butoxy)carbonylamino]-3-chlorophenyl}-N-methylcarbamoyl)-5-oxopyrrolidin-3-yl]methanesulfonate (11.40 g, 20.36 mmol) was dissolved in N-methylpyrrolidone (100 ml) and potassium phosphate (20.07 g, 94.55 mmol) was added. The reaction mixture was stirred at 60° C. for 16 hours. The reaction was monitored by LCMS and TLC (petroleum ether / ethyl acetate = 1:1). After completion of the reaction, the mixture was diluted with water (100 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic layers were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 1:1) to give tert-butyl (11aS, 3aR)-6-chloro-10-methyl-2,11-dioxybenzo[b]pyrrolidino[2,3-f]1,4-diazoindene-5-carboxylate (6,10 g, 75%) as a white solid.

[0213] LCMS(ESI):[M+H] + =380.2.

[0214] Step 9: Dissolve 5-chloro-7-(trifluoromethylthio)-1,3-thiazolino[5,4-b]pyridine (520 mg, 1.54 mmol) in 1,4-dioxane (15.0 mL). Add tert-butyl(11aS,3aR)-6-chloro-10-methyl-2,11-dioxo-4H,11aH,3aH-benzo[b]pyrrolo[2,3-f]1,3-f]1,4-diaminobenzyl (11aS,3aR), potassium carbonate (636 mg, 4.61 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (177.85 mg, 0.31 mmol), and tris(dibenzylideneacetone)dipalladium (141 mg, 0.15 mmol). The mixture was stirred at 100°C for 2 hours. LCMS indicated approximately 30% product. The reaction solution was directly mixed with 100-200 mesh silica gel and purified by normal phase column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain a crude product with a purity of 60%. The product was further purified by reverse phase flash chromatography under the following conditions (C18 column; mobile phase, 30% to 60% acetonitrile / ammonium bicarbonate, detector, UV 254 nm) to obtain tert-butyl (11aS, 3aR)-6-chloro-10-methyl-2,11-dioxo-1-[7-(trifluoromethylethylsulfanyl)(1,3-thiazolinyl[4,5-e]pyridin-5-yl]pyridin-5-yl]benzo[b]pyrrolidino[2,3-f]1,4-diazopiperazine-5-carboxylate (240 mg, 23%) as a white solid.

[0215] LCMS(ESI):[M+H] + =614.3.

[0216] Step 10: Dissolve tert-butyl (3aR, 11aS)-6-chloro-10-methyl-2,11-dioxo-1-(7-(trifluoromethyl)thio)thiazolo[5,4-b]pyridin-5-yl)-1,2,3,3a,4,10,11,11-octahydro-5H-benzo[b]pyrrolo[2,3-f][1,4]diazidine-5-carboxylate (160 mg) in dichloromethane (3.0 mL) and add trifluoroacetic acid (1.0 mL) at 0°C. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored for completion by LCMS. The reactant was concentrated at low temperature, and ethyl acetate (30 mL) and aqueous sodium bicarbonate solution were added to adjust the pH to 8. The mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give (3aR, 11aS)-6-chloro-10-methyl-1-(7-(trifluoromethyl)thio)thiazol[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione (100 mg, 51%) as a white solid.

[0217] LCMS(ESI):[M+H] + =514.1.

[0218] 1 H NMR(400MHz,DMSO-d6)δppm 9.52(s,1H),8.33(s,1H),7.55-7.31(m,3H),5.41(d,J=9.2Hz,1H),4.44-4.43(m,1H),3.60-3.57(m,1H),3.00(s,3H),2.80-2.48(m,4H).

[0219] Step 11: ((3aR,11aS)-6-chloro-10-methyl-1-(7-(trifluoromethyl)thio)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione (60 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (2.0 mL). 3-Bromo-1-propene (70 mg, 0.58 mmol), sodium carbonate (61 mg, 0.58 mmol), and tetrabutylammonium bromide (3.7 mg, 0.01 mmol) were added. The reaction was stirred at 100°C for 16 hours. After overnight reaction, LCMS showed the disappearance of starting material and 85% product. The reactant was poured into water (20 mL) and extracted with ethyl acetate (30 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give (3aR, 11aS)-5-allyl-6-chloro-10-methyl-1-(7-(trifluoromethyl)thio)thiazolo[5,4-b]pyridin-5-yl)-1,3a,4,5,10,11-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione (48 mg, 74%) as a white solid.

[0220] LCMS(ESI):[M+H] + =554.1.

[0221] Step 12: (11aS,3aR)-6-chloro-10-methyl-5-propyl-2-enyl-1-[7-(trifluoromethylthio)(1,3-thiazolo[4,5-e]pyridin-5-yl)]benzo[b]pyrrolidin[2,3-f]1,4-diazoindene-2,11-dione (30 mg, 0.05 mmol) was dissolved in tetrahydrofuran (3.0 mL) and water (0.5 mL) at room temperature. Sodium periodate (35 mg, 0.16 mmol) and potassium osmate dihydrate (1.99 mg, 0.01 mmol) were added, respectively. The reaction mixture was stirred at room temperature for 5 hours. LCMS monitoring showed the disappearance of the starting material and no ion current signal for the product was detected. However, the success of the next step confirmed that the main peak was the product. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The mixture was washed with 0.1 M aqueous sodium thiosulfate (10 mL). The combined organic layers were then dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford crude 2-{(11aS,3aR)-6-chloro-10-methyl-2,11-dioxo-1-[7-(trifluoromethylthio)(1,3-thiazolo[4,5-e]pyridin-5-yl)]benzo[b]pyrrolidinone[2,3-f]1,4-diazolidin-5-yl}acetaldehyde (25.00 mg, 83%), which was used in the next step without further purification.

[0222] LCMS(ESI):[M+H] + =554.1.

[0223] Step 13: Dissolve 2-{(11aS,3aR)-6-chloro-10-methyl-2,11-dioxo-1-[7-(trifluoromethylthio)(1,3-thiazolino[4,5-e]pyridin-5-yl)]benzo[b]pyrrolidino[2,3-f]1,4-diazolidin-5-yl}ethanamine (25.0 mg, 0.04 mmol) in methanol (2.0 mL). Add 1-methylpiperazine (9.0 mg, 0.09 mmol) and acetic acid (1 drop). Stir the reaction at room temperature for 0.5 hours. Add sodium cyanoborohydride (4.2 mg, 0.07 mmol) and continue stirring for 0.5 hours. Monitor the reaction by LCMS, which shows the disappearance of starting material and approximately 75% product. The reaction was added to water (0.5 mL) and directly purified by reverse phase (C18, 0.5% FA / MeCN=5 / 95) and preparative HPLC to give a white solid (11aS, 3aR)-6-chloro-10-methyl-5-[2-(4-methylpiperazinyl)ethyl]-1-[7-(trifluoromethylthio)(1,3-thiazolinyl[4,5-e]pyridin-5-yl)]benzo[b]pyrrolidinyl[2,3-f]1,4-diazoindene-2,11-dione (15.97 mg, 53%).

[0224] LCMS(ESI):[M+H] + =640.3.

[0225] 1 H NMR(400MHz,DMSO-d6)δppm 9.52(s,1H),8.31(s,1H),7.56-7.44(m,3H),5.31(d,J=9.2Hz,1H),3.52-3.51(m,1H), 3.23.25(m,1H),3.00(s,3H),3.00-2.98(m,2H),2.59-2.57(m,1H),2.52-2.10(m,15H).

[0226] 19 FNMR (400MHz, DMSO-d6) δppm-39.06 (s, 3F).

[0227] Example 22: (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thio)thiazol[5,4-b]pyridin-5-yl)-1,3a,4,5,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione

[0228] A similar method as in Example 15 was used to obtain (3aR, 11aS)-6,10-dimethyl-5-(2-(4-methylpiperazin-1-yl)ethyl)-1-(7-(trifluoromethyl)thio)thiazo[5,4-b]pyridin-5-yl)-1,3a,4,5,11a-hexahydro-2H-benzo[b]pyrrolo[2,3-f][1,4]diazonium-2,11(3H)-dione (10.05 mg, 35%).

[0229] LCMS (ESI): 620.20 [M+H] +

[0230] The following compounds were obtained by similar methods to the above examples.

[0231] Effect Example 1

[0232] Polθ polymerase activity assay

[0233] The PicoGreen assay was used to determine the ability of compounds to inhibit Polθ polymerase activity in vitro.

[0234] The His-TEV-SUMO-tagged Polθ protein (amino acids 1792-2590) expressed in E. coli was purified and stored in aliquots at -80°C.

[0235] The assay buffer composition was 25 mM Tris HCl pH 7.5, 12.5 mM NaCl, 0.5 mM MgCl2, 5% glycerol, 0.01% Triton X-100, 0.01% BSA and 1 mM DTT.

[0236] Test compounds were prepared by diluting them in 100% DMSO and using a Bravo (Agilent) to dilute them three-fold to 10 concentrations in a dilution plate (Greiner-781280). The DMSO-diluted compounds were then diluted 20-fold into assay buffer using a Bravo. The Bravo was then used to transfer 2 μl of the diluted compounds to an assay plate (Corning-4512). Purified Polθ enzyme and primers (primer strand: 5'-GCGGCTGTCATAAG-3'): template strand: 5'-GCTACATTGACAATGGCATCAAATCTCAGATTGCGTCTTATGACAGCCGCG-3') (1:1.1) were prepared in assay buffer to a 2.5x working concentration (1.5 nM Polθ and 50 nM PTD). 4 μl was transferred to each well of the assay plate using an E1-CLIPTIP 12-channel electrophoresis system (Thermo, 1-30 μl) and incubated at room temperature for 30 minutes. dNTPs (Sigma-D7295) were diluted in assay buffer to a 2.5x working concentration (40 uM dNTPs) and transferred to each well of the assay plate using an E1-CLIPTIP 12-channel electrophoresis system (Thermo, 1-30 μl) (final DMSO concentration of 1%) and incubated at room temperature for 60 minutes. The reaction was terminated by adding 6 μl of a mixture containing 10 mM EDTA, 25 mM Tris pH 7.5, and a 1:200 dilution of PicoGreen dye (Invitrogen-P7581) to each well of the assay plate. After incubation at room temperature for 30 minutes in the dark, fluorescence was read on an EnVision 2105 (PerkinElmer) using the Ex480nm Em520nm program. The raw data were analyzed using XLfit to generate IC50 values. The results are shown in the table below.

Claims

1. A compound having the structure of formula (I), a pharmaceutically acceptable salt thereof, an isotope isomer, a stereoisomer, in, R 1 Indicates -LR L ; Among them, R 2 represents hydrogen, deuterium, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, deuterated C1-C6 alkyl, halogen, -OR a 、-SR a 、-P(O)R a R b 、CN、Nitro、-S(O)2R a 、-S(O)R a 、-SF5、-NR a R b , halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, halogenated C1-C6 alkylthio, hydroxy C1-C6 alkyl, -C(O)R a 、-C(O)NR a R b 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b or is selected from 0-4 of the following substituents: deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy C1-C6 alkyl, -NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b Replaced by C3-C 10 Cycloalkyl, C6-C 10 Cycloalkenyl, 4-10 membered heterocycloalkyl, 6-10 membered heterocycloalkenyl, C6-C 10 Aryl, 5-10 membered heteroaryl; L represents a bond or a C1-C6 alkylene group, a halogenated C1-C6 alkylene group, wherein any alkylene group may be replaced by -NR a , -O-, -C(O)-, -C(O)O-, -OC(O)-, -NR a C(O)-、-C(O)NR a -, -S- replaced; R L represents a ring B or ring CY-ring D group; or -LR L represents -C1-C6 alkyl or -C1-C6 alkyl-NR a R b wherein the alkyl group is optionally replaced by one or more C1-C6 alkyl groups, C2-C6 alkenyl groups, -NR a , -C(O)-, -C(O)O-, -OC(O)-, -NR a C(O)-、-C(O)NR a -, replaced by S; Ring B represents a C3-C6 cycloalkyl, a 3-6 membered heterocyclyl or a 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2, or 3 members selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent; Ring C represents C3-C6 cycloalkyl, 3-6 membered heterocyclyl or 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2, 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent; Ring D represents C3-C6 cycloalkyl, 3-6 membered heterocyclyl or 5-10 membered heteroaryl, which is optionally substituted by 0, 1, 2, 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent; Wherein, Y represents absence or C1-C6 alkylene; Wherein, R 3 represents hydrogen or C1-C6 alkyl; wherein m and n each independently represent an integer of 0, 1, 2, 3, or 4; Wherein, Cy represents a 9-10 membered saturated or unsaturated fused ring; and, further, the Cy may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; and, further, the Cy may be further selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, R a , R b Each independently represents hydrogen or C1-C6 alkyl.

2. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The Cy represents the following group: Wherein, in Formula I-1, Formula I-2 and Formula I-3: Ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-P(O)R a R b 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; X4 means CR X4 or N; X5 means CR X5 or N; Among them, R X1 , R X2 , R X3 , R X4 , R X5 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b .

3. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The Cy represents the following group: Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; Among them, R X1 , R X2 , R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b ; Wherein, X4 represents CH or N; X5 represents CH or N.

4. The compound of formula (I) according to claim 3, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The Cy represents the following group: Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; Among them, R X1 , R X2 , R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b ; Here, X4 and X5 each independently represent CH or N.

5. The compound of formula (I) according to claim 4, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Cy represents the following group: Furthermore, the Cy may be arbitrarily replaced by 0, 1, 2, or 3 deuterated radicals, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; Among them, R X1 , R X2 , R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, nitro, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b .

6. The compound of formula (I) according to claim 5, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X1 represents CH or N; preferably CH.

7. The compound of formula (I) according to claim 5, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X3 represents CH or N; preferably N.

8. The compound of formula (I) according to claim 5, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X2 means CR X2 or N; where R X2 represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy(C1-C6 alkyl), -NR a R b , CN, halogenated C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b .

9. The compound of formula (I) according to claim 8, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Cy represents the following group:

10. The compound of formula (I) according to claim 3, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The Cy represents the following group: Wherein, ring A represents a saturated or unsaturated ring; and the ring A may contain 0, 1, 2, 3, or 4 heteroatoms selected from O, N, S, and Se; further, the ring A may be arbitrarily replaced by 0, 1, 2, or 3 heteroatoms selected from deuterated, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; Among them, R X1 , R X2 , R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b ; Here, X4 and X5 each independently represent CH or N.

11. The compound of formula (I) according to claim 10, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The Cy represents the following group: Furthermore, the Cy may be arbitrarily replaced by 0, 1, 2, or 3 deuterated radicals, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b substituted by a substituent; Among them, X1 represents CR X1 or N; X2 means CR X2 or N; X3 means CR X3 or N; Among them, R X1 , R X2 , R X3 Each independently represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b .

12. The compound of formula (I) according to claim 11, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X1 represents CH or N; preferably CH.

13. The compound of formula (I) according to claim 11, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X3 represents CH or N; preferably N.

14. The compound of formula (I) according to claim 11, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: X2 means CR X2 or N; where R X2 represents hydrogen, deuterium, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, -OR a , oxo, hydroxy (C1-C6 alkyl), NR a R b , CN, nitro, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, -SO3R a 、-SR a 、-SF5、-C(O)R a 、-C(O)OR a 、-OC(O)R a 、-OC(O)NR a R b 、-NR a COR b Or -CONR a R b .

15. The compound of formula (I) according to claim 11, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Cy represents the following group:

16. The compound of formula (I) according to claim 11, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: L represents -C1-C6 alkylene or -(C1-C6 alkylene)C(O)-, -C(O)(C1-C6 alkylene)-, -(C1-C6 alkylene)-(5-6 membered heteroaryl).

17. The compound of formula (I) according to claim 16, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: L represents -CH2- or -CH2CH2-.

18. The compound of formula (I) according to claim 16, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: L represents -CH2C(O)- or -C(O)CH2-.

19. The compound of formula (I) according to claim 16, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: L represents -CH2-(5-6-membered heteroaryl)-.

20. The compound of formula (I) according to claim 16, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: The 5-6 membered heteroaryl group is selected from any one of the following groups:

21. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: R L represents ring B, wherein ring B represents any of the following groups: Wherein, the ring B is optionally substituted by 0, 1, 2, or 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, -COR a , C1-C6 alkylamino substituent; wherein the wavy line indicates the site of connection to L.

22. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: R L represents ring B, wherein ring B represents any of the following groups: The wavy line indicates the site where L is connected.

23. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: R L represents ring CY-ring D, wherein ring C represents The wavy line indicates the site connected to L, the * indicates the site connected to Y when Y is present, and the * indicates the site connected to ring D when Y is absent.

24. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Y represents absence or -CH2-.

25. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Ring D indicates Wherein, when Y is present, * indicates the site of connection with Y, and when Y is absent, * indicates the site of connection with ring C, wherein the ring D is optionally substituted by 0, 1, 2, or 3 selected from CN, halogen, -NR a R b 、-S(O)2R a , C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxyl, C1-C6 alkoxy, -COR a , substituted by a C1-C6 alkylamino substituent.

26. The compound of formula (I) according to claim 25, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: Ring D indicates 27. The compound of formula (I) according to claim 1, its pharmaceutically acceptable salt, isotope isomer, stereoisomer, wherein: R 1 express 28. A compound having the following structure:

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