An inhibitor having affinity for the bromodomain of smarca2 protein, and preparation method and application thereof

Abstract

The application provides a compound with SMARCA2 protein bromodomain binding ability and a preparation method and application thereof. Specifically, a compound with the structure of formula (I), an enantiomer, a diastereoisomer, a racemate or a pharmaceutically acceptable salt thereof is provided, which is used as an inhibitor with SMARCA2 protein bromodomain binding ability, and is used for preparing a medicine for preventing or treating a disease related to the activity or expression amount of SMARCA2 and SMARCA4 proteins.

Description

Technical Field

[0001] This invention belongs to the field of drug synthesis, specifically relating to an inhibitor with affinity for the bromine domain of the SMARCA2 protein, its preparation method, and its application. Background Technology

[0002] The SWI / SNF chromatin remodeling complexes were first identified in yeast, where they influence mating type transition (SWI) and sucrose nonfermenting (SNF) phenotypes. Since this discovery, homologs of these multi-component protein complexes have been found in other eukaryotes, indicating strong evolutionary conservation of SWI / SNF. In mammals, three SWI / SNF complexes have been identified: BAF, PBAF, and the recently discovered atypical BAF (ncBAF or GBAF). These complexes initiate gene expression by opening tightly packed chromatin structures within the nucleus, allowing transcription factors and their associated proteins to bind. The activity of these complexes depends on the catalytic activity of ATPases, which are provided in a mutually exclusive manner by the SMARCA2 or SMARCA4 proteins. The remaining protein subunits in each SWI / SNF complex can be selected from nearly 30 proteins to determine the biological function of each multi-component complex.

[0003] In various human cancers, researchers have discovered abnormal SWI / SNF complexes in cancer cells. These anomalies are currently attracting considerable attention to explore whether the unusual characteristics of these SWI / SNF complexes in cancer cells can serve as a breakthrough for developing cancer therapeutics. For example, cancer cells carrying SMARCA4 loss-of-function mutations are highly dependent on SMARCA2 to maintain their growth and proliferation. These observations have prompted researchers to actively develop inhibitors of SMARCA2 ATPase activity. Genetic studies have shown that loss of SMARCA4, or a dual loss of SMARCA2 / 4, may have adverse effects on the health of normal cells and tissues. Consistent with these findings, in pharmacodynamic studies of mouse xenograft tumor models, SMARCA2 / 4 ATPase inhibitors, due to a lack of homologous subtype selectivity, exhibit poor tolerability in animal models and lack sufficient biosafety window.

[0004] The C-terminal region of the SMARCA2 protein contains a unique bromine domain, which could serve as another strategy for targeting SMARCA2 in the treatment of SMARCA4-mutant cancers. In recent years, highly effective small-molecule binders / inhibitors of many proteins containing bromine domains have been reported, and some of these compounds have entered clinical trials. Several research teams have reported different series of high-affinity binding molecules (inhibitors) targeting the SMARCA2 bromine domain. However, biological studies on these inhibitors have shown that although potent inhibitors block the binding of the bromine domain in SMARCA2 to histones, their activity in inhibiting cancer cell proliferation is not particularly ideal. In contrast, intracellular knockdown of the SMARCA2 protein using biological methods significantly inhibits the growth and proliferation of SMARCA4-mutant cancer cells. These results suggest that complete degradation of SMARCA2 is an effective strategy for achieving more desirable anticancer efficacy.

[0005] PROTAC technology (protein degradation-targeting chimera) is a rapidly developing protein-targeting degradation technology in recent years. PROTAC molecules can induce the formation of a ternary complex between the target protein and the E3 ubiquitin ligase, leading to ubiquitination of the target protein and subsequent degradation by the proteasome, achieving pharmaceutical "knockdown" or "elimination" of the protein. Several published scientific studies have disclosed PROTAC degraders targeting SMARCA2, demonstrating good antitumor efficacy in SMARCA4 mutant cancer cells and animal models. Therefore, developing inhibitors with SMARCA2 bromine domain binding ability, and subsequently developing SMARCA2 PROTAC degraders, holds significant potential for drug development. Summary of the Invention

[0006] The purpose of this invention is to provide a ligand or inhibitor with affinity for the bromine domain of SMARCA2 protein and / or a degrading agent with SMARCA2 protein degradation activity, its stereoisomer, enantiomer or pharmaceutically acceptable salt thereof, its preparation method and use.

[0007] In a first aspect of the invention, a compound, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof, is provided, wherein the compound is as shown in Formula I;

[0008]

[0009] In the formula,

[0010] R1 is selected from the following group: H, halogens, and hydroxyl groups;

[0011] R2 is the ligand portion of the E3 ubiquitin ligase, or R A L1 ;

[0012] R A L1 Selected from the following group: H, halogen, cyano, optionally substituted C1-C4 alkyl, -OR A P -N(R3)R A P -N(R3)C(O)R A P -C(O)R A P -C(O)OR A P -OC(O)R A P -C(O)N(R3)R A P Optionally substituted C3-C10 carbocyclic groups, Optionally substituted 3- to 10-membered heterocyclic groups, Optionally substituted C6-C10 aryl groups, Optionally substituted 5- to 10-membered heteroaryl groups, Optionally substituted -C3-C10 carbocyclic groups -N(R3)R A P -Optionally substituted -3 to 10-membered subheterocyclic group -N(R3)R A P -Optionally substituted -C6-C10 arylene-N(R3)R A P -Optionally substituted -5 to 10-membered heteroaryl-N(R3)R A P -Optionally substituted -C3-C10 subcarbonyl cyclogroup-OR A P -Optionally substituted -3 to 10-membered heterocyclic group -OR A P -Optionally substituted -C6-C10 arylene-OR A P and - optional substitution of -5 to 10-membered heteroaryl-OR A P ;

[0013] R A P Selected from the following groups: H, C1-C4 alkyl, amino protecting group, hydroxy protecting group;

[0014] R3 is hydrogen or a C1-C4 alkyl group;

[0015] L represents none (single bond) or a connector;

[0016] "Optionally substituted" means that the group is unsubstituted or that one or more hydrogens in the group are replaced by R. S Replaced; each R SIndependently selected from the following group: halogen, -OR a -OC(O)R a -NR a R b -SR a -R a -CN, -NO2, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR b C(O)R a -NR b C(O)2R a -NR a -C(O)NR b R c -S(O)R a -S(O)2R a -S(O)2NR a R b -NR a S(O)2R b and -C1-C2 alkylene-R a ;where R a R b and R c Each is independently selected from the following group: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl, 5-6 heteroaryl, C3-C6 cycloalkyl, 4 to 6 heterocyclic alkyl and benzyl.

[0017] In another preferred embodiment, when used for alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylene, each R S Independently selected from the following group: halogen, -OR a -NR a R b -R a -CN, -NO2, -CO2R a -CONR a R b 、 and -C1-C2 alkylene-R a ;where R a R b and R cEach is independently selected from the following group: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl and C3-C6 cycloalkyl.

[0018] In another preferred embodiment, when used for alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylene, each R S Independently selected from the following group: halogen, -OR a -NR a R b -R a -CN, -NO2; where R a and R b Each is independently selected from the group consisting of: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl and C1-C4 cycloalkyl.

[0019] In another preferred embodiment, when used for alkyl, alkenyl, alkynyl, alkylene, alkenylene, and alkynylene, each R S Independently selected from the group consisting of: D, halogens, and C1-C4 alkyl groups.

[0020] In another preferred embodiment, when used for alkylene, alkenylene, and yntylide groups in L1, each R S Independently selected from the group consisting of: hydrogen, deuterium, halogen, hydroxyl, C1-C4 alkyl, C1-C4 alkoxy, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, and -CN.

[0021] In another preferred embodiment, when used for rings (such as carbocyclic, heterocyclic, aryl, heteroaryl, carbocyclic, heterocyclic, aryl, and heteroaryl), each R S Independently selected from the following group: halogen, -OR a -OC(O)R a -NR a R b -SR a -R a -CN, -NO2, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR b C(O)R a -NR b C(O)2R a -NR a -C(O)NR b Rc -S(O)R a -S(O)2R a -S(O)2NR a R b -NR a S(O)2R b and -C1-C2 alkylene-R a ;where R a R b and R c Each is independently selected from the following group: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl, 5-6 heteroaryl, C3-C6 cycloalkyl, 4- to 6 heterocyclic alkyl and benzyl.

[0022] In another preferred embodiment, when used for rings (such as carbocyclic, heterocyclic, aryl, heteroaryl, carbocyclic, heterocyclic, aryl, and heteroaryl), each R S Independently selected from the following group: halogen, -OR a -OC(O)R a -NR a R b -R a -CN, -NO2, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR b C(O)R a -NR b C(O)2R a -NR a -C(O)NR b R c 、 and -C1-C2 alkylene-R a ;where R a R b and R c Each is independently selected from the following group: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl, C3-C6 cycloalkyl, phenyl, 5-6 membered heteroaryl, C3-C6 cycloalkyl, 4-6 membered heterocycloalkyl and benzyl.

[0023] In another preferred embodiment, when used for rings (such as carbocyclic, heterocyclic, aryl, heteroaryl, carbocyclic, heterocyclic, aryl, and heteroaryl), each RS Independently selected from the group consisting of: deuterium, halogen, hydroxyl, C1-C4 alkoxy, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, -NH2, -NH(C1-C4 alkyl), -N(C1-C4 alkyl)2, -CN, C3-C6 cycloalkyl, phenyl, 5-6 membered heteroaryl, C3-C6 cycloalkyl, 4-6 membered heterocyclic alkyl and benzyl.

[0024] In another preferred embodiment, the linker refers to the divalent linker group in PROTAC used for covalently linking the target protein affinity moiety (R1) and the E3 ubiquitin ligase ligand moiety (R2) for targeting the target protein (i.e., the SMARCA2 protein).

[0025] In another preferred embodiment, R A L1 Selected from the following group: H, halogen, cyano, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, -OR A P (such as C1-C4 alkoxy groups), -N(R3)R A P -N(R3)C(O)R A P (such as C1-C4 carbonyl amine group), -C(O)R A P -C(O)OR A P (such as C1-C3 alkoxycarbonyl), -OC(O)R A P -C(O)N(R3)R A P .

[0026] In another preferred embodiment, R A L1 Selected from the following group: H, halogen, cyano, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 carbonylamino, C1-C3 alkoxycarbonyl.

[0027] In another preferred embodiment, R2 is hydrogen, or a linker fragment having E3 ubiquitinase VHL or CRBN binding capability.

[0028] In another preferred embodiment, R2 is the E3 ubiquitin ligase ligand moiety.

[0029] In another preferred embodiment, the E3 ubiquitin ligase ligand portion is derived from a group based on VHL or CRBN ligands.

[0030] In another preferred embodiment, the E3 ubiquitin ligase ligand portion is a linker fragment based on VHL or CRBN binding ability.

[0031] In another preferred embodiment, the E3 ubiquitin ligase ligand portion (i.e., when R2 is an E3 ubiquitin ligase ligand portion, R2) is selected from the group consisting of:

[0032]

[0033]

[0034]

[0035] In each formula, X1 is selected from the group consisting of hydrogen, halogen, C1-C6 alkoxy, cyano (CN), and optionally substituted C1-C6 alkyl.

[0036] In another preferred embodiment, the optionally substituted C1-C6 alkyl group includes: C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 deuteralkyl, and C1-C6 hydroxyalkyl.

[0037] In another preferred embodiment, X1 is hydrogen, halogen, OMe, CN, or a C1-C3 alkyl substituent.

[0038] In another preferred embodiment, L is a divalent linker or linker.

[0039] In another preferred embodiment, L is R. B L1 -L1.

[0040] In another preferred embodiment, R B L1 Selected from the following group: none, optionally substituted C3-C10 carbocyclic, optionally substituted 3- to 10-membered heterocyclic, optionally substituted C6-C10 arylene, optionally substituted 5- to 10-membered heterocyclic, optionally substituted -C3-C10 carbocyclic-N(R3)-, optionally substituted -3- to 10-membered heterocyclic-N(R3)-, optionally substituted -C6-C10 arylene-N(R3)-, optionally substituted -5- to 10-membered heterocyclic-N(R3)-, optionally substituted -C3-C10 carbocyclic-O-, optionally substituted -3- to 10-membered heterocyclic-O-, optionally substituted -C6-C10 arylene-O- and optionally substituted -5- to 10-membered heterocyclic-O-.

[0041] In another preferred embodiment, R B L1 Selected from the following group:

[0042]

[0043]

[0044] in,

[0045] R3 is hydrogen or a C1-C4 alkyl group;

[0046] m, n, m1, m2, n1, and n2 can each independently represent 1, 2, 3, or 4 (preferably, m+n≤5, m1+n1≤5, and m2+n2≤5).

[0047] In another preferred embodiment, L1 is a divalent linker.

[0048] In another preferred example, L is selected from the following group:

[0049]

[0050] Among the various types,

[0051] m, n, m1, m2, n1, and n2 each independently represent 1, 2, 3, or 4 (preferably, m+n≤5, m1+n1≤5, and m2+n2≤5);

[0052] R3 is hydrogen or a C1-C4 alkyl group;

[0053] L1 is a divalent linker.

[0054] In another preferred embodiment, L1 is selected from the group consisting of: optionally substituted C1-C12 alkylene groups, optionally substituted C1-C12 alkenyl groups, and optionally substituted C1-C12 ynylene groups.

[0055] Furthermore, the -CH2- group in the alkylene, alkenylene, and alkyne groups is independently surrounded by 0, 1, 2, 3, or 4 M groups. a and 0 or 1 M b Replaced; among them,

[0056] M a Each person independently selects from the following groups:

[0057] -O-, -CO-, -NR7-, -CONR7-, -R7NCO-, -NHCONH-, -S-, -SO-, SO2-,

[0058]

[0059] M b Independently selected from the following subcyclic groups: optionally substituted C3-C10 carbon rings, optionally substituted C6-C10 aromatic rings, optionally substituted 3- to 10-membered heterocycles and optionally substituted 5- to 10-membered heteroaromatic rings;

[0060] The condition for attachment is: when there are two -CH2- particles being M aWhen substitution occurs, the two substituted -CH2- groups are separated by at least one C atom (preferably a C atom located on an alkylene chain, an alkenylene chain, or an alkynylene chain), and when both CH2 groups are substituted by groups selected from -O-, -NR7-, and -S-, the two substituted CH2 groups are separated by at least two C atoms (preferably C atoms located on an alkylene chain, an alkenylene chain, or an alkynylene chain).

[0061] Each R7 is independently hydrogen or C1-C3 alkyl.

[0062] In another preferred embodiment, in L1, the optional substitution refers to unsubstituted or one or more H groups being substituted by substituents selected from the group consisting of: D, halogens, C1-C4 alkyl groups.

[0063] In another preferred embodiment, in L1, the alkylene group is a straight-chain alkylene group.

[0064] In another preferred embodiment, in L1, the subalkenyl group is a linear subalkenyl group.

[0065] In another preferred embodiment, in L1, the alkyne group is a straight-chain alkyne group.

[0066] In another preferred embodiment, M b Independently selected from the group consisting of: optionally substituted C3-C7 carbon rings (preferably saturated carbon rings), optionally substituted benzene rings, optionally substituted 4- to 7-membered heterocycles (preferably saturated heterocycles) and optionally substituted 5- to 6-membered heteroaromatic rings.

[0067] In another preferred embodiment, M b The optional subcyclic group selected from the following group:

[0068]

[0069] Where p1 and p2 are each independently 1, 2, 3 or 4 (preferably p1+p2≤5).

[0070] In another preferred embodiment, in L1, the -CH2- in the alkylene, alkenylene, and yntyl groups is independently controlled by 0, 1, 2, 3, or 4 M groups. a What it replaced.

[0071] In another preferred embodiment, in L1, the -CH2- in the alkylene, alkenylene, and yntylide groups is independently separated by 0, 1, or 2 M groups. a and 1 M b What it replaced.

[0072] In another preferred embodiment, L1 is selected from the group consisting of: optionally substituted C1-C12 alkylene groups, optionally substituted C1-C12 alkenyl groups, and optionally substituted C1-C12 ynylene groups.

[0073] Furthermore, one, two, three, or four of the -CH2- groups in the alkylene, alkenylene, and alkyneylene groups are optionally independently selected from the group consisting of groups (i.e., M groups). a Replaced by (group):

[0074] -O-, -CO-, -NR7-, -CONR7-, -R7NCO-, -NHCONH-, -S-, -SO-, SO2-,

[0075]

[0076] The appendix conditions are: when two CH2 groups are substituted, the two substituted -CH2- groups are separated by at least one C atom (preferably, a C atom located on an alkylene chain, an alkenylene chain, or an alkyneylene chain); and when both CH2 groups are substituted by groups selected from -O-, -NR7-, and -S-, the two substituted -CH2- groups are separated by at least two C atoms (preferably, C atoms located on an alkylene chain, an alkenylene chain, or an alkyneylene chain).

[0077] Each R7 is independently hydrogen or a C1-C3 alkyl group;

[0078] The substitution refers to the substitution of one or more H groups by substituents selected from the group consisting of: D, halogens, and C1-C4 alkyl groups.

[0079] In another preferred embodiment, L1 is an optionally substituted divalent linker selected from the group consisting of:

[0080]

[0081] Furthermore, one or two -CH2- groups not within the ring may be independently selected from the group consisting of (i.e., M) a Replaced by (group):

[0082] -O-, -CO-, -NR7-, -CONR7-, -R7NCO-, -NHCONH-, -S-, -SO-, SO2-,

[0083]

[0084] The additional conditions are: when two CH2 atoms are substituted, the two substituted CH2 atoms are separated by at least one C atom, and when both CH2 atoms are substituted by groups selected from -O-, -NR7-, -S-, the two substituted CH2 atoms are separated by at least two C atoms; the substitution refers to one or more H atoms on the group being replaced by groups selected from the group consisting of D and halogens;

[0085] m3 and m4 are each independently 1, 2, 3, 4, 5, 6, 7, 8 or 9 (preferably m3+m4≤11);

[0086] p1 and p2 are each independently 1, 2, 3 or 4 (preferably p1+p2≤5);

[0087] R7 is independently either hydrogen or a C1-C3 alkyl group.

[0088] In another preferred embodiment, the compound is as shown in Formula II;

[0089]

[0090] In the formula, R1, R2 and L are defined as in Formula I.

[0091] In another preferred embodiment, the compound is as shown in Formula III:

[0092]

[0093] In the formula, R1, R2 and L are defined as in Formula I.

[0094] In another preferred embodiment, R1 is a halogen; more preferably, R1 is Br or Cl; even more preferably, R1 is Br or R1 is Cl.

[0095] In another preferred embodiment, the compound is shown in Formula IV;

[0096]

[0097] In the formula, R2 and L are defined as in Formula I.

[0098] In another preferred embodiment, the compound is as shown in Formula V;

[0099]

[0100] In the formula, R2 and L are defined as in Formula I.

[0101] In another preferred embodiment, L1 is selected from the following group:

[0102] -C1-C 12 Alkylene -, -C2-C 12 alkenyl-, -C2-C 12 Ethyne-, -C1-C 12 Alkylene-C3-C 10 Carbon rings -

[0103] -((CH2) s O) t -、-C1-C 12 Alkylene -NH-, -C1-C 12 Alkylene -S-, -C1-C 12 Alkylene-O-

[0104] -C1-C 12 Alkylene-O-C1-C 12 Alkylene-NH-C1-C 12 Alkylene-, C1-C 12 Alkylene S-C1-C 12 Alkylene NH-C1-C 12 Alkylene, C1-C 12 Alkylene O-C1-C 12 Alkylene NH-, C1-C 12 Alkylene S-C0-C 12 Alkylene NH-, C1-C 12 Alkylene PO (C1-C4 alkyl)-C1-C 12 Alkylene NH-, C1-C 12 Alkylene PO (C1-C4 alkyl)-C0-C 12 Alkylene NH, C1-C 12 Alkylene PO (C1-C4 alkyl)-C0-C 12 Alkylene NH-C1-C 12 Alkylene, C1-C 12 alkylene 3-10-membered heterocyclic -C0-C 12 Alkylene, C1-C 12 alkylene 3-10 membered heterocyclic -C1-C 12 Alkylene 3-10 membered heterocycles -, C1-C 12 Alkylene C6-C 10 Aromatic ring-C0-C 12 Alkylene, C1-C 12 Alkylene-C3-C 10 carbon ring -C1-C 12 Alkylene-3-10-membered heterocyclic-C0-C 12 Alkylene;

[0105] In this context, each s is independently 1, 2, 3, or 4; and t is 1, 2, 3, 4, or 5.

[0106] Furthermore, the above groups may be optionally substituted by 1-4 substituents selected from the following group: C1-C6 alkyl, =O;

[0107] Preferably, -C1-C 12 alkylene- and / or -C2-C 12 alkenyl- and / or -C2-C 12 The total chain length of the alkynyl group (i.e., the number of carbon atoms on the alkylene, alkenylene, and alkynylene backbones) is ≤12; preferably, ≤10.

[0108] In another preferred embodiment, L1 is a divalent linker selected from C1-C 10 Alkylene, C2-C 10 alkenyl, C2-C 10 Ethyne group, C1-C 10 Alkylene-C3-C6 carbon ring -、-((CH2) s O) t -、C1-C 10 Alkylene NH-, C1-C 10 Alkylene S-, C1-C 10 Alkylene O-, C1-C 10 Alkylene O-C1-C 10 Alkylene NH-C1-C 10 Alkylene, C1-C 10 Alkylene S-C1-C 10 Alkylene NH-C1-C 10 Alkylene, C1-C 10 Alkylene O-C1-C 10 Alkylene NH-, C1-C 10 Alkylene S-C0-C 10 Alkylene NH-, C1-C 10 Alkylene PO (C1-C2 alkyl)-C1-C 10 Alkylene NH-, C1-C 10 Alkylene PO (C1-C2 alkyl)-C0-C 10 Alkylene NH, C1-C 10 Alkylene PO (C1-C4 alkyl)-C0-C 10 Alkylene NH-C1-C 10 Alkylene, C1-C 10 alkylene 4-8 membered heterocyclic -C0-C 10 Alkylene, C1-C 10 Alkyl 4-8 membered heterocyclic -C1-C 10 Alkyl 4-8 membered heterocyclic -C1-C 10 alkylene ring -C0-C 10 Alkylene, C1-C 10 Alkylene-C3-C6 carbide ring-C1-C 10 Alkylene-4-8-membered heterocyclic-C0-C 12 Alkylene; wherein each s is independently 1, 2, 3 or 4; t is 1, 2, 3, 4 or 5;

[0109] The heterocycle has 1, 2 or 3 heteroatoms selected from the group consisting of N, S, and O, and at least one N as a connecting atom;

[0110] The above groups may be optionally substituted by 1-4 substituents selected from the following group: C1-C4 alkyl, =O;

[0111] Preferably, -C1-C 12 alkylene- and / or -C2-C 12 alkenyl- and / or -C2-C 12 The total chain length of the alkynyl group (i.e., the number of carbon atoms on the alkylene, alkenylene, and alkynylene backbones) is ≤12; preferably, ≤10.

[0112] In another preferred embodiment, L1 is selected from the following group:

[0113]

[0114]

[0115] In another preferred embodiment, R1, R2, L; X1; R B L1 ,L1,R3,m,n,m1,n1,m2,n2,R7,M a M b p1, p2, m3 and m4 are each independently the corresponding groups in the compounds of the embodiments of this application (such as the compounds of Examples 1 to 119 or the compounds in Table A or Table 1).

[0116] In another preferred embodiment, the compound is selected from Table 1.

[0117] In another preferred embodiment, the compound is selected from Table A below:

[0118] Table A

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130]

[0131]

[0132]

[0133] In a second aspect of the invention, a pharmaceutical composition is provided comprising (i) a compound as described in the first aspect, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

[0134] In a third aspect of the invention, the use of a compound as described in the first aspect, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in the second aspect, is provided for the preparation of (i) an inhibitor having the ability to bind to the SMARCA2 protein; and / or (ii) a medicament for the prevention or treatment of diseases related to the activity or expression levels of the SMARCA2 and SMARCA4 proteins.

[0135] In another preferred embodiment, the disease associated with the activity or expression level of the SMARCA2 and SMARCA4 proteins is a disease associated with abnormal activation of SMARCA2.

[0136] In another preferred embodiment, the disease associated with the activity or expression level of SMARCA2 and SMARCA4 proteins is a SMARCA4 loss-of-function disease.

[0137] In another preferred embodiment, the SMARCA4 loss-of-function disease depends on SMARCA2 protein activity.

[0138] In another preferred embodiment, the disease is selected from the group consisting of: cancer, fibrotic diseases.

[0139] In another preferred embodiment, the cancer is selected from the group consisting of: lung cancer, meningioma, mesothelioma, fibrosarcoma, bladder cancer, ovarian cancer, adenocarcinoma, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, bone cancer, brain cancer, neurocytoma, rectal cancer, colon cancer, familial adenomatous polyposis, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, and kidney cancer. Renal parenchymal carcinoma, ovarian cancer, cervical cancer, uterine endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary tract cancer, melanoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma; the fibrotic diseases mentioned are pulmonary fibrosis, liver fibrosis, renal fibrosis, or myocardial fibrosis.

[0140] In a fourth aspect of the invention, a ligand is provided, wherein the ligand is a compound of formula A, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof.

[0141]

[0142] Among them, R1 and R A L1 As defined in the first aspect.

[0143] In another preferred embodiment, R A L1 Selected from the following group:

[0144]

[0145] Where m, n, m1, n1, m2, n2, R3 and R A P As defined in the first aspect.

[0146] In another preferred embodiment, the compound of formula A is shown as formula A1 or A2;

[0147]

[0148] In another preferred embodiment, the compound of formula A is shown as formula A3 or A4;

[0149]

[0150] In another preferred embodiment, R1; R A L1 R3, m, n, m1, n1, m2 and n2 are each independently the corresponding groups in the compounds of the embodiments of this application (such as compounds 1 to 16, 88 and 116-119).

[0151] In another preferred embodiment, the ligand is selected from compounds 1 to 16, 88 and 116-119 (i.e. compounds L-1 to L-21).

[0152] In another preferred embodiment, the ligand is a SMARCA2 protein bromine domain binding ligand.

[0153] In another preferred embodiment, the ligand is used to target the SMARCA2 protein.

[0154] In another preferred embodiment, the ligand is used to prepare a conjugate that targets or binds to the bromine domain of the SMARCA2 protein.

[0155] In another preferred embodiment, the conjugate is a SMARCA2 protein inhibitor; more preferably, the conjugate is a SMARCA2 protein degrader.

[0156] In another preferred embodiment, the conjugate is PROTAC or LYTAC (lysosome-targeting chimaeras).

[0157] In a fifth aspect of the invention, a degrading agent is provided, wherein the degrading agent is a conjugate formed by linking the ligand described in the fourth aspect with an E3 ubiquitinase ligand portion.

[0158] In another preferred embodiment, the ligand is a conjugate formed by linking the ligand to a portion of the E3 ubiquitinase ligand via a linker.

[0159] In another preferred embodiment, the connector is L1, and L1 is as defined above.

[0160] In another preferred embodiment, the ligand is connected to R at one end. A L1 Or R A P Or R A L1 The linker at the location is connected to the E3 ubiquitination enzyme ligand to form a conjugate.

[0161] In another preferred embodiment, the ligand is connected at one end to R. A P The linker at the location is connected to the E3 ubiquitination enzyme ligand to form a conjugate.

[0162] In another preferred embodiment, the degrading agent is a compound of formula B, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof;

[0163]

[0164] Wherein, R2 is the E3 ubiquitin ligase ligand moiety; and the E3 ubiquitin ligase ligand moiety, R1, R B L1 L1 is defined in the first aspect.

[0165] In another preferred embodiment, the compound of formula B is as shown in formula B1 or B2;

[0166]

[0167] In another preferred embodiment, the compound of formula B is as shown in formula B3 or B4;

[0168]

[0169] In another preferred embodiment, R1, R2; X1; R B L1 ,L1,R3,m,n,m1,n1,m2,n2,R7,M a M b p1, p2, m3 and m4 are each independently the corresponding groups in the compounds of the embodiments of this application (such as compounds 17 to 87 and 89 to 115).

[0170] In another preferred embodiment, the degrading agent is selected from compounds 17 to 87 and compounds 89 to 115 (i.e., compounds D-1 to D-98).

[0171] In a sixth aspect of the invention, a pharmaceutical composition is provided comprising (i) a degrading agent as described in the fifth aspect; and (ii) a pharmaceutically acceptable carrier.

[0172] In a seventh aspect of the invention, the use of the degrading agent as described in the fifth aspect or the pharmaceutical composition as described in the sixth aspect is provided in the preparation of (i) SMARCA2 protein inhibitors and / or (ii) medicaments for the prevention or treatment of diseases related to the activity or expression levels of SMARCA2 and SMARCA4 proteins.

[0173] In another preferred embodiment, the diseases associated with the activity or expression level of the SMARCA2 and SMARCA4 proteins are as defined above.

[0174] In an eighth aspect of the invention, a method for degrading SMARCA2 protein is provided, comprising the steps of contacting a compound as described in the first aspect, or an enantiomer, diastereomer, racemate thereof, or a pharmaceutically acceptable salt thereof, or a degrading agent as described in the fifth aspect, with a target to thereby degrade the SMARCA2 protein.

[0175] In another preferred embodiment, the method is an in vitro, non-therapeutic method.

[0176] In another preferred embodiment, the object is the SMARCA2 protein or a cell expressing the SMARCA2 protein.

[0177] In a ninth aspect of the invention, a method for inhibiting cell proliferation is provided, comprising the steps of contacting a compound as described in the first aspect, or an enantiomer, diastereomer, racemate thereof, or a pharmaceutically acceptable salt thereof, or a degrading agent as described in the fifth aspect, with a target to inhibit cell proliferation.

[0178] In another preferred embodiment, the method is an in vitro, non-therapeutic method.

[0179] In another preferred embodiment, the object is a cell expressing the SMARCA2 protein; more preferably, it is a tumor cell.

[0180] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Attached Figure Description

[0181] Figure 1 The results of the degradation of SMARCA2, SMARCA4 and PBRM1 by the compounds of Example 6 are shown.

[0182] Figure 2 The antitumor efficacy of the compound of Example 7 in a mouse model is shown. Detailed Implementation

[0183] Through long-term and in-depth research, the inventors of this invention unexpectedly developed a novel small molecule compound with excellent affinity for the SMARCA2 bromine domain. Building upon this, the inventors further synthesized a bifunctional molecule possessing both SMARCA2 bromine domain binding fragments and E3 ubiquitinase binding capabilities. Through structural modification, design optimization, and activity screening, a class of compounds was ultimately provided that can degrade SMARCA2 protein intracellularly, inhibit cancer cell proliferation, and exhibit excellent anti-cancer activity, potentially applicable to the prevention and treatment of certain cancers. Based on these findings, the inventors completed this invention.

[0184] the term

[0185] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0186] As used herein, when referring to a specific enumerated value, the term “about” means that the value can vary by no more than 1% from the enumerated values. For example, as used herein, the expression “about 100” includes all values ​​between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0187] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed. In other words, the terms also include “consistently made of” or “composed of”.

[0188] As used herein, the term “room temperature” or “normal temperature” refers to a temperature of 4–40°C, preferably 25 ± 5°C.

[0189] When a substituent is described using a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, -CH2O- includes -OCH2-.

[0190] In this invention, the halogen is F, Cl, Br or I.

[0191] In this invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0192] In this invention, the terms "C1-C12" refer to having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms, "C1-C4" refers to having 1, 2, 3, or 4 carbon atoms, and so on. Similarly, "3-6-membered" refers to having 3, 4, 5, or 6 ring atoms, and so on. Similarly, C0 refers to having 0 carbon atoms, i.e., none or none. Therefore, C0-C12 represents either none or having 1 to 12 carbon atoms.

[0193] In this invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon group, wherein the alkyl group used herein, or as part of other groups, has 1 to 12 carbon atoms (i.e., C1-C12 alkyl), 1 to 9 carbon atoms (i.e., C1-C9 alkyl), 1 to 6 carbon atoms (i.e., C1-C6 alkyl), 1 to 4 carbon atoms (i.e., C1-C4 alkyl), or 1 to 3 carbon atoms (i.e., C1-C3 alkyl). Non-limitingly included are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl; preferably ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0194] In this invention, the term "alkoxy" refers to an -O-(alkyl) group. For example, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms (including 1 to 3 carbon atoms), and includes, without limitation, methoxy, ethoxy, n-propoxy, isopropoxy, and butoxy.

[0195] In this invention, the term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing at least one (e.g., 1, 2, or 3) double bonds, for example, the term "C2-C 12 "Alkenyl" refers to a straight-chain or branched alkenyl group having 2 to 12 carbon atoms (including 2-6 carbon atoms or 2-4 carbon atoms) and containing one or two double bonds, including vinyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl, etc.

[0196] In this invention, the term "alkynyl" refers to a straight-chain or branched alkynyl group containing one (e.g., 1, 2, or 3) triple bond, for example, the term "C2-C". 12 "Alynyl" refers to a straight-chain or branched alkynyl group with 2 to 12 carbon atoms (including 2-6 carbon atoms or 2-4 carbon atoms) containing one or two triple bonds. It includes, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentyynyl (2-methyl-3-butynyl, 2-pentynyl, 3-pentynyl) and hexynyl, etc.

[0197] Divalent groups such as "alkylene", "alkenylene", and "alkynylene" refer to divalent groups formed by the loss of one H atom from alkyl, alkenyl, or alkynyl groups. The alkyl, alkenyl, or alkynyl groups defined above in this article can be used for alkylene, alkenylene, and alkynylene, respectively.

[0198] In this invention, the term "carbocyclic group" refers to a fully saturated or unsaturated non-aromatic (e.g., containing one or two double bonds) cyclic hydrocarbon group having a specified number of carbon atoms. The cyclic hydrocarbon group can be monocyclic, bridged, or spirocyclic. For example, C3-C10 carbocyclic groups refer to cyclic hydrocarbon groups having 3 to 10 ring carbon atoms, preferably having 3 to 6 (e.g., 3, 4, 5, or 6) ring carbon atoms, i.e., (C3-C6 carbocyclic groups). When the carbocyclic group is fully saturated, the carbocyclic ring can also be referred to as a cycloalkyl group.

[0199] In this invention, the term "subcarbocyclic group" refers to a divalent group formed by the loss of a hydrogen atom from a carbocyclic group as defined above.

[0200] In this invention, the term "cycloalkyl" refers to a saturated monocyclic, bridged, or spirocyclic cyclic hydrocarbon moiety. For example, the term "C3-C8 cycloalkyl" refers to a monocyclic, bridged, or spirocyclic cyclic alkyl group having 3 to 8 carbon atoms on the ring. Similarly, the term "C3-C6 cycloalkyl" refers to a monocyclic, bridged, or spirocyclic cyclic alkyl group (preferably monocyclic) having 3 to 8 carbon atoms on the ring. Non-limitingly, it includes cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl, etc. The term "bridged cycloalkyl" non-limitingly includes bicyclo[1,1,1]pentyl, bicyclo[2,1,1]hexyl, bicyclo[2,2,1]heptyl, bicyclo[2,2,2]octyl, bicyclo[3,2,2]octyl, etc.

[0201] In this invention, the term "aryl" refers to a hydrocarbon moiety comprising one or more aromatic rings. For example, the term "C6-C..." 10 "Aryl" refers to an aromatic cyclic group with 6 to 10 carbon atoms that does not contain heteroatoms on the ring, such as phenyl and naphthyl.

[0202] In this invention, the term "aryl" refers to a divalent group formed by the loss of a hydrogen atom from an aryl group as defined above.

[0203] In this invention, the term "heterocyclic group" refers to a saturated or unsaturated, non-aromatic (e.g., containing one or two double bonds) cyclic group comprising at least one (e.g., 1, 2, 3, or 4) cyclic heteroatoms (e.g., N, O, or S). The heterocyclic group may have 3 to 10 ring atoms (i.e., a 3 to 10-membered heterocyclic group), preferably, it may have 3 to 6 (e.g., 3, 4, 5, or 6) ring atoms (i.e., a 3 to 6-membered heterocyclic group). Examples of heterocyclic groups include, for example, tetrahydropyridyl, pyrrolinyl, dihydropyridyl, dihydrofuranyl, dihydrothiophenyl, and morpholinyl.

[0204] In this invention, the term "subheterocyclic group" refers to a divalent group formed by the loss of a hydrogen atom from a heterocyclic group as defined above.

[0205] In this invention, the term "heteroaryl" refers to an aromatic cyclic group comprising at least one (e.g., 1, 2, 3, or 4) cyclic heteroatoms (e.g., N, O, or S). Heteroaryl groups may have 5 to 10 cyclic atoms (i.e., 5 to 10-membered heteroaryl groups), preferably, the heterocyclic group may have 5 or 6 cyclic atoms (i.e., 5 to 6-membered heteroaryl groups). Examples of heteroaryl groups include, for example, furanyl, pyrroleyl, thiopheneyl, oxazolyl, imidazolyl, thiazolyl, pyridinyl, quinolinyl, isoquinolinyl, indoleyl, pyrimidinyl, and pyranyl.

[0206] The term "multiple" can refer to 2, 3, 4, 5, or 6.

[0207] In this invention, the term "optionally substituted" means that the group is unsubstituted or substituted by one or more (e.g., 1, 2, 3, and 4) substituents. Unless otherwise defined herein, the substituents may be selected from the group consisting of: -halogens, -OR a -OC(O)R a -NR a R b -SR a -R a -CN, -NO2, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR b C(O)R a -NR b C(O)2R a -NR a -C(O)NR b R c -NH-C(NH2)=NH, -NR a C(NH2)=NH、-NH-C(NH2)=NR a -S(O)R a -S(O)2R a -S(O)2NR a R b -NR a S(O)2R b -N3; where R a R b and R c Independently selected from: hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Deuterated alkyl, C 1-6 aminoalkyl, C 3-6 cycloalkyl, C 2-4 alkenyl, C 2-4 Alkynyl, unsubstituted aryl and heteroaryl, unsubstituted -C 1-4 alkylene-aryl or heteroaryl and unsubstituted -C 1-4 Alkylene-aryloxy compounds, etc.

[0208] The compounds of the present invention

[0209] As used herein, the term "compound of the invention" refers to the compound described in the first aspect of the invention, such as the compound represented by formula (I). The term also includes various crystalline forms of the compound, pharmaceutically acceptable salts, hydrates, or solvates.

[0210] In one aspect, the present invention provides a compound of formula (I), its enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof.

[0211]

[0212] The definitions of R1, R2, and L1 are the same as above.

[0213] In other respects, "compound of the present invention" may also refer to the ligands as described in the fourth aspect of the present invention or the degrading agents as described in the fifth aspect of the present invention.

[0214] As used in this article, "degrading agent" refers to a class of compounds that can target and induce protein degradation through the ubiquitination-proteasome pathway. Generally, such degrading agents are bifunctional small molecules formed by linking a ligand of the target protein (such as SMARCA2 protein) to an E3 ubiquitin ligase ligand, for example, through a linker. They can simultaneously bind to both the target protein and the E3 ubiquitin ligase, causing the target protein to approach the E3 ubiquitin ligase and be ubiquitinated, thereby being recognized and degraded by the intracellular proteasome.

[0215] Salts that may form from the compounds of this invention are also within the scope of this invention. Unless otherwise stated, compounds of this invention are understood to include their salts. The term "salt" as used herein refers to a salt formed from an inorganic or organic acid and a base in an acidic or basic form. Furthermore, when a compound of this invention contains a basic segment, it includes, but is not limited to, pyridine or imidazole; when it contains an acidic segment, it includes, but is not limited to, carboxylic acids; and any zwitterions ("internal salts") that may form are included within the scope of the term "salt." Pharmaceutically acceptable (i.e., non-toxic and physiologically acceptable) salts are preferred, although other salts are also useful, for example, for separation or purification steps in the preparation process. Compounds of this invention may form salts, for example, by reacting compound I with a certain amount, such as an equimolar amount, of an acid or base, precipitating it in a medium, or by freeze-drying it in an aqueous solution.

[0216] The compounds of this invention contain basic fragments, including but not limited to amines, pyridines, or imidazole rings, which may form salts with organic or inorganic acids. Typical acids that can form salts include acetates (such as acetic acid or trihaloacetic acids, such as trifluoroacetic acid), adipates, alginates, ascorbic acid salts, aspartate salts, benzoates, benzenesulfonates, hydrogen sulfates, borates, butyrates, citrates, camphor salts, camphor sulfonates, cyclopentanepropionate, diethylene glycol salts, dodecyl sulfates, ethanesulfonates, fumarates, glucono-2-phosphates, glycerol phosphates, hemisulfates, heptarates, hexanoates, hydrochlorides, hydrobromide, and hydroiodide. Salts, hydroxyethanesulfonates (e.g., 2-hydroxyethanesulfonate), lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g., 2-naphthalenesulfonate), nicotinates, nitrates, oxalates, pectates, persulfates, phenylpropionates (e.g., 3-phenylpropionates), phosphates, picrates, neopentanoates, propionates, salicylates, succinates, sulfates (e.g., those formed with sulfuric acid), sulfonates, tartrates, thiocyanates, toluenesulfonates such as p-toluenesulfonate, dodecanoates, etc.

[0217] Some compounds of this invention may contain acidic fragments, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases. Typical base-formed salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, and salts formed with organic bases (such as organic amines), such as benzathine, dicyclohexylamine, hepatoamine, N-methyl-D-glucosamine, N-methyl-D-glucosamide, tert-butylamine, and salts formed with amino acids such as arginine, lysine, etc.

[0218] This invention is also intended to include crystal forms of the compounds of this invention, such as hydrates, solvates, etc.

[0219] Some compounds of this invention may exist in specific geometric or stereoisomeric forms. This invention covers all compounds, including their cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures, and other mixtures. Additionally, the asymmetric carbon atom may represent a substituent, such as an alkyl group. All isomers and mixtures thereof are included in this invention.

[0220] According to the present invention, the ratio of isomers in a mixture of isomers can be varied. For example, a mixture containing only two isomers can have the following combinations: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0. All ratios of isomers are within the scope of the present invention. Similar ratios readily understood by those skilled in the art, as well as ratios for mixtures of more complex isomers, are also within the scope of the present invention.

[0221] This invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. However, in practice, it is common for one or more atoms to be replaced by atoms with different atomic weights or mass numbers. Examples of isotopes that can be included in the compounds of this invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, respectively as follows: 2 H, 3 H, 13 C 11 C 14 C 15 N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. The compounds of this invention, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates, wherein the isotopes or other isotopic atoms of the aforementioned compounds are all within the scope of this invention. Certain isotopically labeled compounds of this invention, for example... 3 H and 14 Radioactive isotopes of carbon are also included, and are useful in tissue distribution experiments of drugs and substrates. Tritium, i.e. 3 H and carbon-14, i.e. 14 C, their preparation and detection are relatively easy. They are the preferred isotopes. In addition, heavier isotopes such as deuterium are used for substitution. 2 H, due to its excellent metabolic stability, offers advantages in certain therapies, such as increasing half-life or reducing dosage in vivo, and therefore may be preferred in some cases. Isotopically labeled compounds can be prepared using general methods, by replacing the non-isotopic reagent with an readily available isotopically labeled reagent, according to the scheme described in the examples.

[0222] Pharmaceutical compositions and their uses

[0223] The present invention provides a pharmaceutical composition comprising the compound of the present invention or its enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof; and a pharmaceutically acceptable carrier.

[0224] The compounds of this invention possess SMARCA2 protein bromine domain affinity and / or SMARCA2 protein degradation activity, and therefore can be used to prepare medicaments (such as SMARCA2 protein degraders) for the prevention and / or treatment of diseases related to the activity or expression level of SMARCA2 / 4 proteins, including cancer and fibrotic diseases. The cancers include, but are not limited to, lung cancer, meningioma, mesothelioma, fibrosarcoma, bladder cancer, ovarian cancer, adenocarcinoma, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, bone cancer, brain cancer, neurocytoma, rectal cancer, colon cancer, familial adenomatous polyposis, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, renal cancer, renal parenchymal carcinoma, and ovarian cancer. Cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary tract cancer, melanoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, multiple myeloma; the fibrotic diseases mentioned include, but are not limited to, pulmonary fibrosis, liver fibrosis, kidney fibrosis, or myocardial fibrosis.

[0225] The compounds of the present invention (especially the degrading agents of the present invention) can be used in combination with other known drugs for treating or improving similar conditions. When used in combination, the administration method and dosage of the original drugs can remain unchanged, while the compound of the present invention is taken simultaneously or subsequently. When the compound of the present invention is taken concurrently with one or more other drugs, a pharmaceutical composition containing one or more known drugs and the compound of the present invention is preferred. Drug combination also includes taking the compound of the present invention with one or more other known drugs during overlapping time periods. When the compound of the present invention is used in combination with one or more other drugs, the dosage of the compound of the present invention or the known drugs may be lower than the dosage of either drug alone. The pharmaceutical composition may also contain one or more anticancer agents, for example, selected from (but not limited to): PARP1 / 2 inhibitors, chemotherapeutic agents that induce DNA damage in cancer cells, DNA alkylation agents, DNA or RNA synthesis inhibitors, EGFR, ALK, or FGFR tyrosine receptor kinase inhibitors, KRAS, MEK, or ERK tumor signaling pathway inhibitors, and tumor immunotherapy drugs (such as PD-1 antibodies, PD-L1 antibodies, etc.).

[0226] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that facilitate the formulation and / or administration and / or absorption of the active agent by an individual and can be included in the compositions disclosed herein without causing significant adverse toxicological effects on that individual. Non-limiting examples of pharmaceutically acceptable carriers and excipients include water, NaCl, physiological saline solutions, lactated Ringer's solution, conventional sucrose, conventional glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, and pigments. Such formulations can be sterilized and, if desired, mixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts affecting osmotic pressure, buffers, colorants, and / or aromatic substances that will not harmfully react with or interfere with the activity of the compounds provided herein. Those skilled in the art will recognize that other pharmaceutical carriers and excipients are suitable for the disclosed compounds.

[0227] In some embodiments, the pharmaceutical compositions of the present invention may be in solid or liquid form.

[0228] The pharmaceutical product containing the active ingredient of this invention can be a suitable oral dosage form, such as tablets, pills, lozenges, water-soluble or oily suspensions, dispersed latex powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Orally administered pharmaceutical products can be prepared according to known process methods of the pharmaceutical ingredient manufacturer. These compositions may include one or more of the following agents: sweeteners, flavoring agents, coloring agents, and protective agents, to provide an elegant and palatable pharmaceutical formulation. The tablet contains the active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for tablet production. Examples of such excipients include inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating agents, disintegrants such as corn starch or alginic acid; binding agents such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. The tablet may be uncoated or coated to delay degradation and absorption in the gastrointestinal tract, thereby maintaining its activity over a longer period.

[0229] The active compound may be administered to a subject via any suitable route, including orally, parenterally, by inhalation spray, locally, rectally, nasally, sublingually, vaginally, or via an implantable cartridge. As used herein, the term "parenterally" includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrasheathic, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the composition is administered orally, intraperitoneally, or intravenously.

[0230] Pharmaceutical compositions suitable for use in this invention are typically in the form of discrete units in solid form, such as tablets, capsules, pouches, powders, granules, lozenges, patches, suppositories, pills, or in liquid form, such as liquid formulations, injectable or infusionable solutions or suspensions.

[0231] The precise amount of compound provided to an individual for therapeutic efficacy will depend on the route of administration, the type and severity of the disease and / or condition, and individual characteristics such as general health, age, sex, weight, and tolerance to the drug. Those skilled in the art will be able to determine the appropriate dosage based on these and other factors. When administered in combination with other therapeutic agents, the “therapeutic efficacy” of any other therapeutic agent will depend on the type of drug used. Appropriate dosages are known for approved therapeutic agents and can be adjusted by those skilled in the art based on the individual’s condition, the type of condition being treated, and the amount of the compound of the invention used below, for example, the dosages reported in the literature and recommended in Physician's Desk Reference (57th edition, 2003). Preferably, the compositions should be formulated such that an inhibitory dose of 0.01-100 mg / kg body weight / day can be administered to patients receiving these compositions. In some embodiments, the compositions of the invention provide doses from 0.01 mg to 50 mg. In other embodiments, doses of 0.1 mg-25 mg or 5 mg-40 mg are provided.

[0232] Examples of subjects to which the pharmaceutical compositions or therapeutic agents of the present invention are administered include mammals (e.g., humans, mice, rats, hamsters, rabbits, cats, dogs, cattle, sheep, monkeys, etc.).

[0233] The present invention also provides a treatment method comprising the steps of: administering to a subject requiring treatment a compound of the present invention, an enantiomer, a diastereomer, a racemic mixture or a pharmaceutically acceptable salt thereof, or administering a pharmaceutical composition of the present invention to block the binding between the bromine domain / histone of the SMARCA2 protein, or to degrade the SMARCA2 protein, or to treat a disease associated with a loss-of-function mutation of SMARCA4.

[0234] Preparation method

[0235] The compounds of the present invention can be prepared by the following reaction route.

[0236] Route 1: The route for preparing compounds (I-1) and (I-2)

[0237]

[0238] Step 1: Dissolve compound S1 and substituted benzaldehyde compound S2 in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and add a sodium alkoxide organic base (sodium methoxide, sodium ethoxide, sodium tert-butoxide). Then, heat the reaction solution at 40-90°C for an appropriate time to obtain the final product S3.

[0239] Step 2: Compound S4 and substituted benzaldehyde compound S5 are dissolved in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added. The reaction solution is then stirred at room temperature for an appropriate time to obtain the final product S6.

[0240] Step 3: Add excess hydrazine hydrate aqueous solution to either S3 or S6. Then, reflux the reaction solution for an appropriate time to obtain the final product S7.

[0241] Step 4: Mix S7 with carbon tetrachloride, acetonitrile, or 1,2-dichloroethane, and then add NBS and AIBN respectively. Afterward, reflux the reaction solution for an appropriate time to obtain the final product S8.

[0242] Step 5: Mix S8 with an inert solvent (e.g., acetonitrile, toluene, 1,2-dichloroethane, 1,4-dioxane), add an alkali (potassium carbonate, sodium carbonate, K3PO3, etc.), and heat the reaction solution at 70-90°C for an appropriate time to obtain the final product S9.

[0243] Step 6: First add DMSO to S9 (R=F), then add piperazine and alkali (potassium carbonate, sodium carbonate, K3PO3, etc.), heat at 100-140℃ for an appropriate time to obtain the final product (I)-1.

[0244] Step 7: Mix S9 (R=Br) and S10, then add an inert solvent (e.g., 1,4-dioxane, ethylene glycol dimethyl ether, toluene, etc.) and water. Next, add a base (potassium carbonate, sodium carbonate, K3PO3, triethylamine, etc.) to remove oxygen. Finally, add a palladium catalyst (e.g., Pd(dppf)Cl2-CH2Cl2, Pd(PPh3)4, Pd2(dba)3, PdCl2, Pd(OAc)2, etc.). Heat the reaction solution at 70-120℃ for an appropriate time to obtain the final product S11.

[0245] Step 8: Add S11 to an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, acetonitrile, etc.), mix well, add HCl solution of 1,4-dioxane, stir the reaction solution at room temperature for an appropriate time to obtain the final product (I)-2.

[0246] Route 2: The route for preparing compounds (I-3), (I-4), and (I-5)

[0247]

[0248] Step 1: Known compounds S20 and S21, or S20 and S22, are mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.). Then, a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added, and the reaction solution is allowed to react at room temperature to yield one of S24.

[0249] Under other conditions, S23 is mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, DBU, etc.) and a condensing agent (diphenylphosphine chloride, CDI, PyBOP, HATU, and EDCI, etc.) are added. After the reaction, amine S20 is added. Finally, the reaction solution reacts at room temperature to give an S24.

[0250] Step 2: S24 is mixed in an inert solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) and a condensing agent (diphenylphosphine chloride, CDI, PyBOP, HATU, and EDCI, etc.) are added. The reaction solution is then allowed to react (e.g., 0.4-2 hours), followed by the addition of (I)-2. The reaction is then carried out at room temperature (e.g., 0.4-6 hours) to yield (I)-3.

[0251] Step 3: Mix S25 and (I)-1, or S25 and (I)-2, in an inert solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and add a base (potassium carbonate, cesium carbonate, triethylamine, ethyl diisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) and potassium iodide (sodium iodide, lithium iodide, tetra-n-butylammonium iodide, etc.). Then, heat the reaction solution at 40-70°C for an appropriate time (e.g., 4-12 hours) to obtain the final products (I)-4 and (I)-5.

[0252] Route 3: The route for preparing compounds (I-6) and (I-7)

[0253]

[0254] Step 1: Compounds S31 and (I)-1 are mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added. Subsequently, at room temperature, the reaction solution is reacted with sodium borohydride acetate as a reducing agent, resulting in a reductive amination reaction to yield the final product (I)-6. Under another condition, S31 and (I)-1 are mixed in an inert solvent, and a weak acid (acetic acid, formic acid, citric acid, etc.) is added. Subsequently, at room temperature, the reaction solution is reacted with sodium borohydride acetate as a reducing agent, resulting in a reductive amination reaction to yield the final product (I)-6.

[0255] Similarly, compounds S31 and (I)-2 are known to be mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added. Subsequently, at room temperature, the reaction solution is subjected to a reducing agent, sodium borohydride acetate, to obtain the final product (I)-7 via a reductive amination reaction. Under other conditions, S31 and (I)-2 are mixed in an inert solvent, and a weak acid (acetic acid, formic acid, citric acid, etc.) is added. Subsequently, at room temperature, the reaction solution is subjected to a reducing agent, sodium borohydride acetate, to obtain the final product (I)-7 via a reductive amination reaction.

[0256] Route 4: The route for preparing compounds (I-8), (I-9), (I-10), and (I-11)

[0257]

[0258] Step 1: Compounds S41 and (I)-1 or S42 and (I)-1 are mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added. Subsequently, the reaction solution is kept at room temperature, and the reducing agent sodium borohydride acetate is added, resulting in a reductive amination reaction to yield the final products (I)-8 and (I)-10.

[0259] Under other conditions, S41 and (I)-1 or S42 and (I)-1 are mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a weak acid (acetic acid, formic acid, citric acid, etc.) is added. Subsequently, the reaction solution is kept at room temperature, and the reducing agent sodium borohydride acetate is added to obtain the final products (I)-8 and (I)-10 through a reductive amination reaction.

[0260] Similarly, compounds S41 and (I)-2 or S42 and (I)-2 are known to be mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and then a base (triethylamine, ethyldiisopropylamine, N,N-diisopropylethylamine, and DBU, etc.) is added. Subsequently, the reaction solution is kept at room temperature, and the reducing agent sodium borohydride acetate is added, resulting in a reductive amination reaction to give the final products (I)-9 and (I)-11.

[0261] Under other conditions, S41 and (I)-2 or S42 and (I)-2 are mixed in an inert solvent (e.g., methanol, tetrahydrofuran, 2-methyltetrahydrofuran, 1,4-dioxane, toluene, dichloromethane, ethanol, 1,2-dichloroethane, N,N-dimethylformamide, acetonitrile, etc.), and a weak acid (acetic acid, formic acid, citric acid, etc.) is added. Subsequently, the reaction solution is kept at room temperature, and the reducing agent sodium borohydride acetate is added to obtain the final products (I)-9 and (I)-11 through a reductive amination reaction.

[0262] The main advantages of this invention include:

[0263] 1. A novel class of small molecule compounds with affinity for the bromine domain of the SMARCA2 protein is provided. The preparation method has the advantages of mild reaction conditions, abundant and readily available raw materials, and simple operation and post-processing.

[0264] 2. A novel class of small molecule compounds with VHL and CRBN ligands and similar structures, as well as affinity for the bromine domain of the SMARCA2 protein, is provided. These compounds have the function of degrading SMARCA2 in cells. The preparation method has the advantages of mild reaction conditions, abundant and readily available raw materials, and simple operation and post-processing.

[0265] 3. A compound with good anticancer activity against SMARCA4 mutant cancer cells is provided; this type of compound has good solubility, high bioavailability and excellent metabolic properties, and is a potential antitumor drug.

[0266] The invention is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0267] Example 1: Synthesis of key intermediates and target compounds

[0268] (1) Synthesis of key intermediate: (4-bromo-3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate dimethyl ester (LJI149)

[0269]

[0270] Step 1: Synthesis of intermediate methyl 2-(acetoxymethyl)-6-bromobenzoate (LJI068)

[0271] The reactants methyl 2-methyl-6-bromobenzoate (7.4 g, 32 mmol), NBS (6.3 g, 35 mmol), and AIBN (530 mg, 3.2 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 2 hours. After the reaction was complete, insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure, redissolved in tetrahydrofuran, cooled to 0 °C, and diethyl phosphite (6.7 g, 48 mmol) and DIEA (6.3 g, 48 mmol) were slowly added. The mixture was then heated to room temperature and stirred for 2 hours. After the reaction was complete, the system was concentrated under reduced pressure, extracted with water and ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain a crude product (9.7 g, 97%), a pale yellow liquid, which was used directly in the next reaction.

[0272] The crude product (9.7 g, 31 mmol), potassium acetate (6.2 g, 63 mmol), and tetrabutylammonium iodide (233 mg, 0.63 mmol) were dissolved in tetrahydrofuran and stirred at room temperature for 12 hours. After the reaction was complete, the solid was removed by filtration, the filtrate was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the target compound LJI068 (7.46 g, 82%) as a pale yellow liquid. 1 ¹H NMR (500MHz, chloroform-d) δ 7.52 (dd, J = 1.0, 8.1Hz, 1H), 7.33 (dd, J = 1.0, 7.7Hz, 1H), 7.23 (t, J = 7.9Hz, 1H), 5.08 (s, 2H), 3.92 (s, 3H), 2.04 (s, 3H). LRMS (ESI) calculated values: C 11 H 12 79 BrO4[M+H] + =287.0; Measured value: 287.0.

[0273] Step 2: Synthesis of intermediate 7-bromoisobenzofuran-1(3H)-one (LJI076)

[0274] LJI068 (7.46 g, 26 mmol) was dissolved in methanol, and potassium carbonate (179 mg, 1.3 mmol) was added. The mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the system was concentrated under reduced pressure, extracted with water and dichloromethane, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain crude LJI076 (5.53 g, 100%), a white solid, which was used directly in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 7.69 (dd, J = 1.0, 7.8Hz, 1H), 7.52 (t, J = 7.7Hz, 1H), 7.44 (dd, J = 1.0, 7.6Hz, 1H), 5.26 (s, 2H). LRMS (ESI) calculated value: C₈H₆ 79 BrO2[M+H] + =213.0; Measured value: 213.0.

[0275] Step 3: Synthesis of intermediate 3,7-dibromoisobenzofuran-1(3H)-one (LJI078)

[0276] LJI076 (5.53 g, 26 mmol), NBS (5.08 g, 29 mmol), and AIBN (427 mg, 2.6 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 2 hours. After the reaction was complete, insoluble matter was removed by diatomaceous earth filtration. The filtrate was concentrated under reduced pressure and purified by silica gel column chromatography to obtain the target compound LJI078 (7.11 g, 93%) as a yellow liquid.1 ¹H NMR (400MHz, chloroform-d) δ 7.77–7.73 (m, 1H), 7.64–7.56 (m, 2H), 7.34 (s, 1H). LRMS (ESI) calculated value: C8H5 79 Br2O2[M+H] + =290.9; Measured value: 290.9.

[0277] Step 4: Synthesis of intermediate 3-hydroxy,7-bromoisobenzofuran-1(3H)-one (LJI148)

[0278] LJI078 (7.11 g, 24 mmol) was dissolved in water and refluxed at 100 °C for 2 hours. After the reaction was complete, the mixture was filtered, and the filter cake was washed successively with water and a small amount of dichloromethane, and then dried to obtain the target compound LJI148 (4.55 g, 81%), which was a white solid. 1 HNMR (400MHz, DMSO-d6) δ 8.19 (d, J = 8.1Hz, 1H), 7.85 (dd, J = 2.0, 6.7Hz, 1H), 7.73–7.64 (m, 2H), 6.60 (d, J = 7.7Hz, 1H). LRMS (ESI) calculated value: C8H6 79 BrO3[M+H] + =229.0; Measured value: 229.0.

[0279] Step 5: Synthesis of dimethyl (4-bromo-3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate (LJI149)

[0280] Sodium methoxide (28 mmol, 5 M in MeOH) was dissolved in methanol and cooled to 0 °C. Then, dimethyl phosphite (2.88 g, 26 mmol) was added, and the mixture was stirred at 0 °C for 10 minutes. LJI148 (4 g, 17 mmol) was dissolved in methanol and slowly added to the reaction system, followed by stirring at room temperature for 1 hour. Finally, methanesulfonic acid (2.85 g, 29 mmol) was added to the system, and the mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the system was concentrated under reduced pressure, extracted with water and ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain crude LJI149 (quantitatively converted), a yellow liquid that did not require purification and could be used directly in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 7.74–7.65 (m, 2H), 7.54 (t, J = 7.8 Hz, 1H), 5.65 (d, J = 11.2 Hz, 1H), 3.91 (d, J = 11.0 Hz, 3H), 3.63 (d, J = 10.5 Hz, 3H). LRMS (ESI) calculated values: C10 H 11 79 BrO5P[M+H] + =321.0; Measured value: 321.0.

[0281] (2) Key intermediate for synthesis: dimethyl (4-chloro-3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate (LJK011)

[0282]

[0283] Step 1: Synthesis of intermediate methyl 2-(acetoxymethyl)-6-bromobenzoate (LJK128)

[0284] Following the synthetic method of LJI068, methyl 2-methyl-6-chlorobenzoate (7.9 g, 43 mmol), NBS (8.4 g, 47 mmol), AIBN (703 mg, 4.3 mmol), diethyl phosphite (8.8 g, 64 mmol), and DIEA (8.3 g, 64 mmol) were used as raw materials to obtain a crude product (10.7 g, 97%). This crude product was then reacted with potassium acetate (8 g, 81 mmol) and tetrabutylammonium iodide (300 mg, 0.81 mmol) to obtain the target compound LJK128 (9.03 g, 91%), which was a yellow liquid.

[0285] Step 2: Synthesis of intermediate 7-chloroisobenzofuran-1(3H)-one (LJK126)

[0286] Following the synthesis method of LJI076, crude LJK126 (5.90 g, 94%) was obtained from LJK128 (9.03 g, 37 mmol) and potassium carbonate (257 mg, 1.86 mmol) as raw materials. It was a white solid and was directly used in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 7.58 (t, J = 7.7Hz, 1H), 7.43 (dd, J = 0.9, 7.8Hz, 1H), 7.39 (dd, J = 0.9, 7.6Hz, 1H), 5.25 (s, 2H). LRMS (ESI) calculated value: C8H6 35 ClO2[M+H] + =169.0; Measured value: 169.0.

[0287] Step 3: Synthesis of intermediate 3-chloro,7-bromoisobenzofuran-1(3H)-one (LJK132)

[0288] Following the synthesis method of LJI078, LJK126 (5.9 g, 35 mmol), NBS (6.8 g, 38 mmol), and AIBN (575 mg, 3.5 mmol) were used as raw materials. The target compound LJK132 (8.6 g, 99%) was obtained by silica gel column purification and was a yellow solid.

[0289] Step 4: Synthesis of intermediate 3-hydroxy,7-chloroisobenzofuran-1(3H)-one (LJI133)

[0290] Following the synthesis method of LJI148, crude LJK133 (6.4 g, 99%) was obtained from LJK132 (8.6 g, 35 mmol) as a raw material. It was a white solid and was directly used in the next reaction.

[0291] Step 5: Synthesis of dimethyl (4-chloro-3-oxo-1,3-dihydroisobenzofuran-1-yl)phosphonate (LJK011)

[0292] Following the synthesis method of LJI149, using LJK133 (1.48 g, 8.1 mmol), sodium methoxide (12.1 mmol, 5 M inMeOH), dimethyl phosphite (1.77 g, 16 mmol), and methanesulfonic acid (1.70 g, 17 mmol) as raw materials, crude product LJK011 (quantitative conversion) was obtained. It was a yellow liquid that did not require purification and could be used directly in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 7.68–7.64 (m, 2H), 7.38 (d, J = 2.0 Hz, 1H), 5.69 (dd, J = 1.0, 11.3 Hz, 1H), 3.95 (d, J = 11.0 Hz, 3H), 3.66 (d, J = 11.0 Hz, 3H). LRMS (ESI) calculated values: C 10 H 11 35 ClO5P[M+H] + =277.0; Measured value: 277.0.

[0293] Synthesized target compound 1: 5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK070)

[0294]

[0295] Step 1: Synthesis of compound 2-(o-tolyl)-1H-inden-1,3(2H)-dione (LJK064)

[0296] Phenylphthalide (250 mg, 1.86 mmol) and o-tolualdehyde (246 mg, 2.05 mmol) were dissolved in ethyl acetate / methanol, and sodium methoxide (7.44 mmol, 5 M in MeOH) was added. The mixture was refluxed at 70 °C for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure. The crude product was diluted with water and adjusted to pH 4 with 1 N hydrochloric acid solution, precipitating a solid. The solid was filtered, and the filter cake was washed successively with water and n-hexane. After drying, crude product LJK064 was obtained as a yellowish-white solid, which was directly used in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 8.11–8.02 (m, 2H), 7.94–7.86 (m, 2H), 7.26–7.19 (m, 2H), 7.13 (td, J = 2.1, 7.3 Hz, 1H), 6.89 (dd, J = 1.2, 7.5 Hz, 1H), 4.52 (s, 1H), 2.33 (s, 3H). LRMS (ESI) calculated values: C 16 H 13 O2[M+H] + =237.1; Measured value: 237.1.

[0297] Step 2: Synthesize compound 4-(2-methylbenzyl)phthalazine-1(2H)-one (LJK066)

[0298] The crude product LJK064 was dissolved in 85% hydrazine hydrate and refluxed at 100°C for 5 hours. After the reaction was completed, the mixture was filtered, the filter cake was washed with water, and dried to obtain LJK066 (324 mg, 69%), which was a light brownish-red solid. 1 ¹H NMR (400MHz, chloroform-d) δ 10.77 (s, 1H), 8.53–8.45 (m, 1H), 7.81–7.73 (m, 2H), 7.73–7.67 (m, 1H), 7.25–7.21 (m, 1H), 7.17 (td, J = 1.4, 7.4 Hz, 1H), 7.09 (td, J = 1.5, 7.5 Hz, 1H), 6.99 (dd, J = 1.3, 7.6 Hz, 1H), 4.28 (s, 2H), 2.39 (s, 3H). LRMS (ESI) calculated values: C 16 H 15 N₂O[M+H] + =251.1; Measured value: 251.1.

[0299] Step 3: Synthesis of compound 5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK070):

[0300] LJK066 (312 mg, 1.25 mmol), NBS (333 mg, 1.87 mmol), and AIBN (20 mg, 0.12 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (170 mg, 1.25 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane. The mixture was dried to obtain the crude product, which was then purified by HPLC to obtain the trifluoroacetate of the target compound, LJK070 (70 mg, 22%), as a white solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.14 (s, ¹H), 9.74 (s, ¹H), 8.98 (d, J = 8.2 Hz, ¹H), 8.58-

[0301] 8.41 (m, 3H), 8.28-8.16 (m, 2H), 8.13-8.03 (m, 2H). LRMS (ESI) calculated values: C 16 H 10 N₂O[M+H] + =246.1; Measured value: 246.1.

[0302] Synthesized target compound 2: 4-chloro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK019)

[0303]

[0304] Step 1: Synthesis of compound (Z)-7-chloro-3-(2-methylbenzylmethylene)isobenzofuran-1(3H)-one (LJK014):

[0305] LJK011 (460 mg, 1.66 mmol) and o-methylbenzaldehyde (200 mg, 1.66 mmol) were dissolved in ultra-dry tetrahydrofuran. Triethylamine (337 mg, 3.33 mmol) was added dropwise with stirring, and the mixture was stirred at room temperature for 12 hours. After the reaction was complete, a solid precipitated. The solvent was removed under reduced pressure, and the mixture was diluted with water. The mixture was stirred at room temperature for 0.5 hours, filtered, and the filter cake was washed successively with water and n-hexane. After drying, crude product LJK014 (320 mg, 71%) was obtained as a yellow solid, which was used directly in the next reaction. 1¹H NMR (400MHz, chloroform-d) δ 8.14 (d, J = 7.7Hz, 1H), 7.71 (d, J = 7.7Hz, 1H), 7.63 (t, J = 7.8Hz, 1H), 7.48 (d, J = 7.7Hz, 1H), 7.31–7.27 (m, 1H), 7.24–7.20 (m, 2H), 6.66 (s, 1H), 2.47 (s, 3H). LRMS (ESI) calculated values: C 16 H 12 35 ClO2[M+H] + =271.1; Measured value: 271.1.

[0306] Step 2: Synthesis of compound 8-chloro-4-(2-methylbenzyl)phthalazine-1(2H)-one (LJK017):

[0307] LJK014 (320 mg, 1.18 mmol) was dissolved in anhydrous ethanol, and 85% hydrazine hydrate (118 mg, 2.36 mmol) was added. The mixture was refluxed at 80 °C for 12 hours. After the reaction was completed, a solid precipitated from the system. The solid was filtered, and the filter cake was washed successively with water, ethanol, and n-hexane. The solid was dried to give (LJK017) (184 mg, 54%), which was a white solid. 1 ¹H NMR (400MHz, DMSO-d⁶) δ 12.51 (s, 1H), 7.90–7.76 (m, 3H), 7.19 (dd, J = 1.5, 7.6Hz, 1H), 7.11 (td, J = 1.5, 7.4Hz, 1H), 7.05 (td, J = 1.6, 7.4Hz, 1H), 6.93 (dd, J = 1.4, 7.6Hz, 1H), 4.23 (s, 2H), 2.30 (s, 3H). LRMS (ESI) calculated values: C 16 H 14 35 ClN2O[M+H] + =285.1; Measured value: 285.1.

[0308] Step 3: Synthesize compound 4-chloro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK019)

[0309] Following the synthesis method of LJK070, LJK017 (184 mg, 0.65 mmol), NBS (173 mg, 0.97 mmol), AIBN (11 mg, 0.06 mmol), and potassium carbonate (90 mg, 0.65 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJK019 (52 mg, 29%), was obtained by HPLC purification and was a yellow solid. 1¹H NMR (400MHz, methanol-d⁴) δ 10.07 (s, ¹H), 9.75 (s, ¹H), 8.96 (dd, J = 3.0, 6.3Hz, ¹H), 8.53 (d, J = 8.4Hz, ¹H), 8.46 (d, J = 8.4Hz, ¹H), 8.22 (ddd, J = 1.2, 6.9, 8.3Hz, ¹H), 8.15–8.03 (m, ³H). LRMS (ESI) calculated values: C 16 H 10 35 ClN2O[M+H] + =281.0; Measured value: 281.0.

[0310] Synthesized target compound 3: 4-bromo-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK002)

[0311]

[0312] Step 1: Synthesize compound (Z)-7-bromo-3-(2-methylbenzylmethylene)isobenzofuran-1(3H)-one (LJK004)

[0313] Following the synthesis method of LJK014, using LJI149 (534 mg, 1.66 mmol), o-methylbenzaldehyde (200 mg, 1.66 mmol), and triethylamine (337 mg, 3.33 mmol) as raw materials, crude product LJK004 (364 mg, 69%) was obtained as a yellow solid, which was directly used in the next reaction. 1 ¹H NMR (400MHz, chloroform-d) δ 8.17–8.11 (m, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.55 (t, J = 7.8 Hz, 1H), 7.31–7.26 (m, 1H), 7.24–7.20 (m, 2H), 6.65 (s, 1H), 2.47 (s, 3H). LRMS (ESI) calculated values: C 16 H 12 79 BrO2[M+H] + =315.0; Measured value: 315.0.

[0314] Step 2: Synthesis of compound 8-bromo-4-(2-methylbenzyl)phthalazine-1(2H)-one (LJI140)

[0315] Following the synthesis method of LJK017, LJK004 (364 mg, 1.15 mmol) and 85% hydrazine hydrate (115 mg, 2.3 mmol) were used as raw materials to obtain LJI140 (200 mg, 52%), which was a white solid. 1 ¹H NMR (400MHz, DMSO-d⁶) δ 12.51 (s, 1H), 8.05 (dd, J = 1.1, 7.8Hz, 1H), 7.85 (dd, J = 1.1, 8.2Hz, 1H), 7.71 (t, J = 8.0Hz, 1H), 7.23–7.16 (m, 1H), 7.11 (td, J = 1.5, 7.4Hz, 1H), 7.05 (td, J = 1.5, 7.5Hz, 1H), 6.93 (dd, J = 1.4, 7.5Hz, 1H), 4.23 (s, 2H), 2.31 (s, 3H). LRMS (ESI) calculated values: C 16 H 14 79 BrN2O[M+H] + =329.0; Measured value: 329.0.

[0316] Step 3: Compound 4-bromo-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJI143)

[0317] Following the synthesis method of LJK070, LJI140 (50 mg, 0.15 mmol), NBS (30 mg, 0.17 mmol), AIBN (3 mg, 0.02 mmol), and potassium carbonate (21 mg, 0.15 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJI143 (20 mg, 41%), was obtained by HPLC purification and was a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.07 (s, ¹H), 9.73 (s, ¹H), 9.00 (dd, J = 1.0, 8.3Hz, ¹H), 8.53 (d, J = 8.5Hz, ¹H), 8.46 (d, J = 8.4Hz, ¹H), 8.35 (dd, J = 1.0, 7.9Hz, ¹H), 8.21 (ddd, J = 1.2, 6.9, 8.4Hz, ¹H), 8.08 (ddd, J = 1.1, 6.9, 8.2Hz, ¹H), 7.99 (t, J = 8.1Hz, ¹H). LRMS (ESI) calculated values: C 16 H 10 79 BrN2O[M+H] + =325.0; Measured value: 325.0.

[0318] Synthesized target compound 4: 11-fluoro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK072)

[0319]

[0320] Steps 1 and 2: Synthesis of compound 4-(5-fluoro-2-methylbenzyl)phthalazine-1(2H)-one (LJK068)

[0321] Following the synthesis method of LJK066, using phthalide (250 mg, 1.86 mmol), 2-methyl-5-fluorobenzaldehyde (257 mg, 1.86 mmol), sodium methoxide (7.44 mmol, 5 M in MeOH), and hydrazine hydrate as raw materials, LJK068 (395 mg, 79%) was obtained as a white solid. 1 ¹H NMR (400MHz, chloroform-d) δ 10.32 (s, 1H), 8.55–8.45 (m, 1H), 7.85–7.73 (m, 2H), 7.71–7.61 (m, 1H), 7.17 (dd, J = 5.9, 8.4Hz, 1H), 6.86 (td, J = 2.8, 8.4Hz, 1H), 6.70 (dd, J = 2.8, 9.7Hz, 1H), 4.24 (s, 2H), 2.35 (s, 3H). LRMS (ESI) calculated values: C 16 H 14 19 FN2O[M+H] + =269.1; Measured value: 269.1.

[0322] Step 3: Synthesis of 11-fluoro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK072)

[0323] Following the synthesis method of LJK070, using LJK068 (395 mg, 1.47 mmol), NBS (393 mg, 2.21 mmol), AIBN (24 mg, 0.15 mmol), and potassium carbonate (200 mg, 1.47 mmol) as raw materials, the trifluoroacetate of the target compound, LJK072 (243 mg, 62%), was purified by HPLC and obtained as a yellow solid. 1¹H NMR (400MHz, methanol-d⁴) δ 10.14 (s, 1H), 9.67 (s, 1H), 8.93 (d, J = 8.1Hz, 1H), 8.63 (dd, J = 5.3, 9.3Hz, 1H), 8.47 (dd, J = 1.3, 8.0Hz, 1H), 8.20 (ddd, J = 1.4, 7.4, 8.3Hz, 1H), 8.16–8.04 (m, 2H), 7.91 (td, J = 2.4, 8.9Hz, 1H). LRMS (ESI) calculated values: C 16 H 10 19 FN2O[M+H] + =265.1; Measured value: 265.1.

[0324] Synthesized target compound 5: 5-oxo-11-(piperazin-1-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK073)

[0325]

[0326] LJK072 (50 mg, 0.19 mmol), anhydrous piperazine (24 mg, 0.28 mmol), and potassium phosphate (60 mg, 0.28 mmol) were dissolved in DMSO and reacted at 120 °C for 1 hour. After the reaction was completed, the crude product was purified by HPLC to obtain the trifluoroacetate of the target compound, LJK073 (35 mg, 56%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.51(s,1H),8.99(s,1H),8.68(d,J=8.1Hz,1H),8.34(dd,J=1.3,7.9Hz,1H),8.18(d,J=9.4Hz,1H),8.10- 8.01(m,1H),8.00-7.92(m,1H),7.75(dd,J=2.4,9.4Hz,1H),7.42(d,J=2.4Hz,1H),3.90(t,J=5.3Hz,4H),3.48(t,J=5.3Hz,4H). 13 C NMR (101MHz, MeOD) δ 162.44, 155.26, 142.83, 139.69, 135.92, 134.68, 133.85, 131.63, 129.56, 126.81, 124.94, 123.78, 122.53, 119.63, 117.97, 106.47, 45.27, 44.18. LRMS (ESI) calculated values: C 20 H 19 N4O[M+H] +=331.2; Measured value: 331.2.

[0327] Synthesized target compound 6: 4-chloro-11-fluoro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG012)

[0328]

[0329] Step 1: Synthesis of compound (Z)-7-chloro-3-(5-fluoro-2-methylbenzylmethylene)isobenzofuran-1(3H)-one (LCG009)

[0330] Following the synthetic method of LJK014, using LJK011 (480 mg, 1.74 mmol), 2-methyl-5-fluorobenzaldehyde (240 mg, 1.74 mmol), and triethylamine (352 mg, 3.47 mmol) as starting materials, crude product LCG009 (248 mg, 49%) was obtained as a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H 11 35 Cl 19 FO2[M+H] + =289.0; Measured value: 289.0.

[0331] Step 2: Synthesis of compound 8-chloro-4-(5-fluoro-2-methylbenzyl)phthalazine-1(2H)-one (LCG010)

[0332] Following the synthesis method of LJK017, LCG009 (260 mg, 0.9 mmol) and 85% hydrazine hydrate (90 mg, 1.8 mmol) were used as raw materials to obtain LCG010 (206 mg, 75%), which was a white solid. 1 ¹H NMR (400MHz, chloroform-d) δ 10.08 (s, 1H), 7.74 (d, J = 7.8Hz, 1H), 7.64 (t, J = 8.0Hz, 1H), 7.52 (d, J = 8.0Hz, 1H), 7.17 (dd, J = 5.9, 8.4Hz, 1H), 6.86 (td, J = 2.7, 8.5Hz, 1H), 6.66 (d, J = 9.6Hz, 1H), 4.18 (s, 2H), 2.34 (s, 3H). LRMS (ESI) calculated values: C 16 H 13 35 Cl 19 FN2O[M+H] + =303.1; Measured value: 303.1.

[0333] Step 3: Synthesize compound 4-chloro-11-fluoro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG012)

[0334] LCG010 (206 mg, 0.68 mmol), NBS (182 mg, 1.02 mmol), and AIBN (11 mg, 0.07 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (94 mg, 0.68 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane, and dried to obtain LCG012 (106 mg, 52%), a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H9 35 Cl 19 FN2O[M+H] + =299.0; Measured value: 299.0.

[0335] Synthesize target compound 7: 4-chloro-5-oxo-11-(piperazin-1-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG013):

[0336]

[0337] LCG012 (106 mg, 0.36 mmol), anhydrous piperazine (37 mg, 0.43 mmol), and potassium phosphate (91 mg, 0.43 mmol) were dissolved in DMSO and reacted at 120 °C for 1 hour. After the reaction was completed, the crude product was purified by HPLC to obtain the trifluoroacetate of the target compound, LCG013 (75 mg, 58%), as a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 9.56 (s, 1H), 9.11 (s, 1H), 8.80–8.68 (m, 1H), 8.27 (d, J = 9.2 Hz, 1H), 8.09–7.94 (m, 2H), 7.86 (d, J = 9.3 Hz, 1H), 7.52 (s, 1H), 3.95 (t, J = 4.9 Hz, 4H), 3.48 (t, J = 4.9 Hz, 4H). LRMS (ESI) calculated values: C 20 H 18 35 ClN4O[M+H] + =365.1; Measured value: 365.0.

[0338] Synthesized target compound 8: 4-bromo-11-fluoro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK049)

[0339]

[0340] Step 1: Synthesize compound (Z)-7-bromo-3-(5-fluoro-2-methylbenzylmethylene)isobenzofuran-1(3H)-one (LJK032)

[0341] Following the synthetic method for LJK014, using LJI149 (348 mg, 1.09 mmol), 2-methyl-5-fluorobenzaldehyde (150 mg, 1.09 mmol), and triethylamine (220 mg, 2.17 mmol) as starting materials, crude product LJK032 (122 mg, 33%) was obtained as a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H 11 79 Br 19 FO2[M+H] + =333.0; Measured value: 333.0.

[0342] Step 2: Synthesize compound 8-bromo-4-(5-fluoro-2-methylbenzyl)phthalazine-1(2H)-one (LJK034)

[0343] Following the synthesis method of LJK017, LJK034 (120 mg, 94%) was obtained as a yellow solid using LJK032 (122 mg, 0.37 mmol) and 85% hydrazine hydrate (37 mg, 0.74 mmol) as raw materials. 1 ¹H NMR (400MHz, chloroform-d) δ 10.29 (s, 1H), 7.99 (dd, J = 1.6, 7.4Hz, 1H), 7.61–7.47 (m, 2H), 7.17 (dd, J = 5.9, 8.5Hz, 1H), 6.85 (td, J = 2.7, 8.4Hz, 1H), 6.64 (dd, J = 2.7, 9.7Hz, 1H), 4.18 (s, 2H), 2.34 (s, 3H). LRMS (ESI) calculated values: C 16 H 13 79 Br 19 FN2O[M+H] + =347.0; Measured value: 347.0.

[0344] Step 3: Synthesize compound 4-bromo-11-fluoro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK049)

[0345] LJK034 (60 mg, 0.17 mmol), NBS (46 mg, 0.26 mmol), and AIBN (3 mg, 0.02 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (24 mg, 0.17 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane, and dried to obtain LJK049 (38 mg, 64%), a yellow solid, which was directly used in the next reaction. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.10 (s, ¹H), 9.70 (s, ¹H), 8.98 (d, J = 8.2 Hz, ¹H), 8.64 (dd, J = 5.2, 9.3 Hz, ¹H), 8.38 (d, J = 7.8 Hz, ¹H), 8.14 (d, J = 8.7 Hz, ¹H), 8.00 (t, J = 8.1 Hz, ¹H), 7.96–7.90 (m, ¹H). LRMS (ESI) calculated values: C 16 H9 79 Br 19 FN2O[M+H] + =343.0; Measured value: 343.0.

[0346] Synthesized target compound 9: 4-bromo-5-oxo-11-(piperazin-1-yl)-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK050)

[0347]

[0348] LJK049 (50 mg, 0.15 mmol), anhydrous piperazine (25 mg, 0.29 mmol), and potassium phosphate (62 mg, 0.29 mmol) were dissolved in DMSO and reacted at 120 °C for 1 hour. After the reaction was completed, the crude product was purified by HPLC to obtain the trifluoroacetate of the target compound, LJK050 (15 mg, 26%), as a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 9.40 (s, 1H), 8.91 (s, 1H), 8.64 (d, J = 8.2Hz, 1H), 8.21–8.10 (m, 2H), 7.82 (t, J = 8.0Hz, 1H), 7.75 (dd, J = 2.3, 9.4Hz, 1H), 7.37 (d, J = 2.4Hz, 1H), 3.92 (t, J = 5.3Hz, 4H), 3.49 (t, J = 5.3Hz, 4H). LRMS (ESI) calculated values: C 20 H 1879 BrN4O[M+H] + =409.1; Measured value: 409.1.

[0349] Synthesized target compound 10: 11-bromo-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-iod-6-ide (LJK095)

[0350]

[0351] Steps 1 and 2: Synthesis of compound 4-(5-bromo-2-methylbenzyl)phthalazine-1(2H)-one (LJK091)

[0352] Following the synthesis method of LJK066, using phthalide (200 mg, 1.49 mmol), 2-methyl-5-bromobenzaldehyde (297 mg, 1.49 mmol), sodium methoxide (5.96 mmol, 5 M in MeOH), and hydrazine hydrate as raw materials, LJK091 (279 mg, 56%) was obtained as a white solid. 1 ¹H NMR (400MHz, chloroform-d) δ 10.24 (s, 1H), 8.56–8.44 (m, 1H), 7.89–7.73 (m, 2H), 7.72–7.63 (m, 1H), 7.30 (dd, J = 2.1, 8.2Hz, 1H), 7.15 (s, 1H), 7.09 (d, J = 8.1Hz, 1H), 4.23 (s, 2H), 2.32 (s, 3H). LRMS (ESI) calculated values: C 16 H 14 79 BrN2O[M+H] + =329.0; Measured value: 329.0.

[0353] Step 3: Synthesize compound 11-bromo-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJK095)

[0354] LJK091 (270 mg, 0.82 mmol), NBS (219 mg, 1.23 mmol), and AIBN (13 mg, 0.08 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (113 mg, 0.82 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane, and dried to obtain LJK095 (147 mg, 55%), a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H 1079 BrN2O[M+H] + =325.0; Measured value: 325.0.

[0355] Synthesized target compound 11: 5-oxo-11-(1,2,3,6-tetrahydropyridin-4-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK100)

[0356]

[0357] LJK095 (74 mg, 0.23 mmol), SM3 (84 mg, 0.27 mmol), Pd(dppf)Cl2-CH2Cl2 (18 mg, 0.02 mmol), and potassium carbonate (47 mg, 0.34 mmol) were dissolved in dioxane / water (volume ratio 10:1). The system was evacuated of air and purged with nitrogen, and reacted at 80 °C for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and extracted with a mixture of water and dichloromethane / methanol. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain crude LJK099, a brownish-black liquid, which was directly used in the next reaction.

[0358] The crude product LJK099 was dissolved in a dichloromethane / methanol mixture, and hydrochloric acid (2.3 mmol, 4 M in Dioxane) was added. The mixture was stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed under reduced pressure. The crude product was washed successively with diethyl ether and dichloromethane, dried, and then purified by HPLC to obtain the trifluoroacetate salt LJK100 (48 mg, 64%) of the target compound, which was a yellow solid. 1 HNMR (500MHz, methanol-d4) δ 10.06 (s, 1H), 9.68 (s, 1H), 8.95 (d, J = 8.2Hz, 1H), 8.54–8.43 (m, 3H), 8.31–8.17 (m, 2H), 8.09 (t, J = 7.6Hz, 1H), 6.74 (s, 1H), 4.03 (s, 2H), 3.61 (t, J = 6.0Hz, 2H), 3.07 (d, J = 7.2Hz, 2H). LRMS (ESI) calculated values: C 21 H 18 N3O[M+H] + =328.1; Measured value: 328.1.

[0359] Synthesized target compound 12: 11-bromo-4-chloro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJM156)

[0360]

[0361] Step 1: Synthesis of compound (Z)-3-(5-bromo-2-methylbenzylmethylene)-7-chloroisobenzofuran-1(3H)-one (LJK051)

[0362] Following the synthetic method for LJK014, using LJK011 (695 mg, 2.51 mmol), 2-methyl-5-bromobenzaldehyde (500 mg, 2.51 mmol), and triethylamine (508 mg, 5.02 mmol) as starting materials, crude product LJK051 (608 mg, 69%) was obtained as a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H 11 79 Br 35 ClO2[M+H] + =349.0; Measured value: 349.0.

[0363] Step 2: Synthesize compound 4-(5-bromo-2-methylbenzyl)-8-chlorophthalazine-1(2H)-one (LJK052)

[0364] Following the synthesis method of LJK017, LJK052 (338 mg, 53%) was obtained from LJK051 (608 mg, 1.74 mmol) and 85% hydrazine hydrate (174 mg, 3.48 mmol) as raw materials, and was a yellow solid. 1 ¹H NMR (400MHz, chloroform-d) δ 10.02 (s, 1H), 7.75 (dd, J = 1.1, 7.8Hz, 1H), 7.66 (t, J = 8.0Hz, 1H), 7.55 (dd, J = 1.1, 8.1Hz, 1H), 7.30 (dd, J = 2.1, 8.1Hz, 1H), 7.12–7.06 (m, 2H), 4.17 (s, 2H), 2.31 (s, 3H). LRMS (ESI) calculated values: C 16 H 13 79 Br 35 ClN2O[M+H] + =363.0; Measured value: 363.0.

[0365] Step 3: Synthesize compound 11-bromo-4-chloro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJM156)

[0366] LJK052 (600 mg, 1.65 mmol), NBS (440 mg, 2.47 mmol), and AIBN (27 mg, 0.16 mmol) were dissolved in carbon tetrachloride and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (456 mg, 3.30 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane, and dried to obtain LJM156 (249 mg, 42%), a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H9 79 Br 35 ClN2O[M+H] + =359.0; Measured value: 359.0.

[0367] Synthesized target compound 13: 4-chloro-5-oxo-11-(1,2,3,6-tetrahydropyridin-4-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK056)

[0368]

[0369] Step 1: Synthesize compound 11-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-4-chloro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL126)

[0370] LJM156 (249 mg, 0.69 mmol), SM3 (204 mg, 0.66 mmol), Pd(dppf)Cl2-CH2Cl2 (28 mg, 0.03 mmol), and potassium carbonate (192 mg, 1.39 mmol) were dissolved in dioxane / water (10:1, v / v). The system was evacuated of air and purged with nitrogen, and reacted at 80 °C for 12 hours. After the reaction was complete, the mixture was concentrated under reduced pressure, and extracted with a mixture of water and dichloromethane / methanol. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain the target compound LJL126 (216 mg, 67%) as a yellow solid. 1 ¹H NMR (400 MHz, chloroform-d) δ 9.18 (s, ¹H), 8.67 (s, ¹H), 8.24 (s, ¹H), 8.00–7.73 (m, ³H), 7.73–

[0371] 7.54 (m, 2H), 6.40 (s, 1H), 4.33–4.03 (m, 2H), 3.82–3.61 (m, 2H), 2.77–2.54 (m, 2H), 1.51 (s, 9H). LRMS (ESI) calculated values: C 26 H 25 35 ClN3O3[M+H] + =462.2; Measured value: 462.2.

[0372] Step 2: Synthesize compound 4-chloro-5-oxo-11-(1,2,3,6-tetrahydropyridin-4-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK056)

[0373] LJL126 (210 mg, 0.46 mmol) was dissolved in a dichloromethane / methanol mixture, and hydrochloric acid (4.6 mmol, 4 min Dioxane) was added. The mixture was stirred at room temperature for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure. The resulting solid was washed successively with diethyl ether and dichloromethane, dried, and then purified by HPLC to obtain the trifluoroacetate salt of the target compound, LJK056 (152 mg, 93%), as a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.02 (s, ¹H), 9.70 (s, ¹H), 8.93 (dd, J = 2.1, 7.3Hz, ¹H), 8.52 (d, J = 8.9Hz, ¹H), 8.46 (s, ¹H), 8.29 (dd, J = 1.7, 8.9Hz, ¹H), 8.17–8.07 (m, 2H), 6.75 (s, ¹H), 4.03 (d, J = 3.4Hz, 2H), 3.61 (t, J = 6.1Hz, 2H), 3.11–3.01 (m, 2H). LRMS (ESI) calculated values: C 21 H 17 35 ClN3O[M+H] + =362.1; Measured value: 362.1.

[0374] Synthesized target compound 14: 4-chloro-11-(1-(4-methoxy-4-oxobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL118)

[0375]

[0376] LJK056 (20 mg, 0.06 mmol), methyl 4-bromobutyrate (20 mg, 0.11 mmol), potassium carbonate (15 mg, 0.11 mmol), and potassium iodide (18 mg, 0.11 mmol) were dissolved in acetonitrile / DMF (1:1, v / v) and reacted at 80 °C for 2 hours. After the reaction was completed, the solution was concentrated under reduced pressure and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL118 (10.7 mg, 41%), as a yellow solid. 1 H NMR (500MHz, methanol-d4) δ9.69 (s, 1H), 9.27 (s, 1H), 8.70 (dd, J = 1.6, 7.8Hz, 1H), 8.23 ​​(d, J=9.0Hz,1H),8.14(s,1H),8.05-7.99(m,2H),7.93(dd,J=1.8,9.0Hz,1H),6.53(s,1 4.41-4.13 (m, 1H), 4.13-3.82 (m, 2H), 3.73 (s, 3H), 3.60-3.45 (m, 1H), 3.44-3.37 (m, 2H), 3.25-2.94 (m, 2H), 2.60 (t, J = 7.0 Hz, 2H), 2.25-2.17 (m, 2H). LRMS (ESI) calculated values: C 26 H 25 35 ClN3O3[M+H] + =462.2; Measured value: 462.2.

[0377] Synthesized target compound 15: 10-bromo-4-chloro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJM025)

[0378]

[0379] Step 1: Synthesis of compound (Z)-3-(4-bromo-2-methylbenzylmethylene)-7-chloroisobenzofuran-1(3H)-one (LJL156)

[0380] Following the synthetic method of LJK014, using LJK011 (770 mg, 2.78 mmol), 2-methyl-4-bromobenzaldehyde (554 mg, 2.78 mmol), and triethylamine (422 mg, 4.18 mmol) as starting materials, crude product LJL156 (700 mg, 71%) was obtained as a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H 11 79 Br 35 ClO2[M+H] +=349.0; Measured value: 349.0.

[0381] Step 2: Synthesis of compound 4-(4-bromo-2-methylbenzyl)-8-chlorophthalazine-1(2H)-one (LJL160)

[0382] Following the synthesis method of LJK017, LJL156 (700 mg, 2.0 mmol) and 85% hydrazine hydrate (200 mg, 4.0 mmol) were used as raw materials to obtain LJL160 (690 mg, 94%), which was a white solid. 1 ¹H NMR (400MHz, chloroform-d) δ 9.68 (s, 1H), 7.75 (dd, J = 1.2, 8.0Hz, 1H), 7.65 (t, J = 8.0Hz, 1H), 7.55 (dd, J = 1.2, 8.1Hz, 1H), 7.38 (d, J = 2.1Hz, 1H), 7.25–7.21 (m, 1H), 6.83 (d, J = 8.1Hz, 1H), 4.15 (s, 2H), 2.34 (s, 3H). LRMS (ESI) calculated values: C 16 H 13 79 Br 35 ClN2O[M+H] + =363.0; Measured value: 363.0.

[0383] Step 3: Synthesize compound 10-bromo-4-chloro-5-oxo-5H-isoquinolinyl[3,2-a]phthalazine-7-onthium-6-oxide (LJM025)

[0384] LJL160 (767 mg, 2.11 mmol), NBS (563 mg, 3.16 mmol), and AIBN (35 mg, 0.21 mmol) were dissolved in 1,2-dichloroethane and refluxed at 80 °C for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product and potassium carbonate (583 mg, 4.22 mmol) were dissolved in acetonitrile and refluxed at 80 °C for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, the system was diluted with water, filtered, and the filter cake was washed successively with water and dichloromethane, and dried to obtain LJM025 (217 mg, 28%), a yellow solid, which was directly used in the next reaction. LRMS (ESI) calculated value: C 16 H9 79 Br 35 ClN2O[M+H] + =359.0; Measured value: 359.0.

[0385] Synthesized target compound 16: 4-chloro-5-oxo-10-(1,2,3,6-tetrahydropyridin-4-yl)-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJM027)

[0386]

[0387] Step 1: LJM025 (253 mg, 0.71 mmol), SM3 (219 mg, 0.71 mmol), Pd(dppf)Cl2-CH2Cl2 (29 mg, 0.04 mmol), and potassium carbonate (195 mg, 1.41 mmol) were dissolved in dioxane / water (volume ratio 10:1). The system was evacuated of air and purged with nitrogen, and reacted at 80 °C for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure. The system was extracted with water and a dichloromethane / methanol mixed solvent. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain crude LJM026, a brownish-black liquid, which was directly used in the next step of the reaction.

[0388] Step 2: The crude product LJM026 was dissolved in a dichloromethane / methanol mixture, and hydrochloric acid (7.1 mmol, 4 min Dioxane) was added. The mixture was stirred at room temperature for 12 hours. After the reaction was completed, the solvent was removed under reduced pressure. The crude product was washed successively with diethyl ether and dichloromethane, dried, and then purified by HPLC to obtain the trifluoroacetate salt LJM027 (93 mg, 36%) of the target compound, which was a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.04 (s, 1H), 9.70 (s, 1H), 8.94 (dd, J = 2.9, 6.4Hz, 1H), 8.52 (s, 1H), 8.49–8.38 (m, 2H), 8.15–8.05 (m, 2H), 6.67 (d, J = 3.6Hz, 1H), 4.01 (d, J = 3.6Hz, 2H), 3.59 (t, J = 6.1Hz, 2H), 3.07–2.99 (m, 2H). LRMS (ESI) calculated values: C 21 H 17 35 ClN3O[M+H] + =362.1; Measured value: 362.1.

[0389] Synthesized target compound 17: 4-chloro-11-(1-(4-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-4-oxobutyryl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL058)

[0390]

[0391] LJK056 (10 mg, 0.03 mmol), known compound LJL057 (15 mg, 0.03 mmol), HATU (12 mg, 0.03 mmol), and DIEA (14 mg, 0.11 mmol) were dissolved in DMF and stirred at room temperature for 1 hour. After the reaction was completed, the system was quenched with water, extracted with a dichloromethane / methanol mixed solvent, concentrated under reduced pressure, and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL058 (4.6 mg, 19%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.91(s,1H),9.61(s,1H),8.95(s,1H),8.92-8.84(m,1H),8.43(d,J=9.3Hz,1H), 8.34(d,J=2.9Hz,1H),8.29-8.21(m,1H),8.13-8.04(m,2H),7.50-7.37(m,4H),6.73(s,1H),4.65-4.4 7(m,4H), 4.45-4.30(m,3H), 3.97-3.82(m,3H), 3.80(dd,J=3.9,11.0Hz,1H), 2.89-2.57(m,6H), 2.47(s,3H), 2.23(dd,J=7.8,13.3Hz,1H), 2.08(ddd,J=4.4,9.1,13.3Hz,1H), 1.04(s,9H). LRMS(ESI) calculated value: C 47 H 49 35 ClN7O6S[M+H] + =874.3; Measured value: 874.3.

[0392] Synthesized target compound 18: 4-chloro-11-(1-(5-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-5-oxopentanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL061)

[0393]

[0394] Following the synthetic method of LJL058, LJK056 (10 mg, 0.03 mmol), known compound LJL059 (15 mg, 0.03 mmol), HATU (12 mg, 0.03 mmol), and DIEA (14 mg, 0.11 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL061 (22.6 mg, 91%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.92(s,1H),9.60(s,1H),9.51(s,1H),8.89(dd,J=2.9,6.5Hz,1H),8.42(d,J=9.0Hz,1H), 8.35(s,1H),8.25(dd,J=1.7,9.0Hz,1H),8.12-8.04(m,2H),7.53-7.42(m,4H),6.74(s,1H),4.65-4.50(m,4H), 4.43-4.32 (m, 3H), 4.00-3.78 (m, 4H), 2.88-2.82 (m, 1H), 2.81-2.72 (m, 1H), 2.60-2.50 (m, 4H), 2.46-2.32 (m, 3H), 2.29-2.21 (m, 1H), 2.10 (ddd, J = 4.4, 9.1, 13.3 Hz, 1H), 2.02-1.91 (m, 2H), 1.09-0.98 (m, 9H). LRMS (ESI) calculated values: C 48 H 51 35 ClN7O6S[M+H] + =888.3; Measured value: 888.3.

[0395] Synthesized target compound 19: 4-chloro-11-(1-(6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-6-oxohexanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL097)

[0396]

[0397] Following the synthesis method of LJL058, LJK056 (10 mg, 0.03 mmol), known compound LJL095 (15 mg, 0.03 mmol), HATU (12 mg, 0.03 mmol), and DIEA (14 mg, 0.11 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL097 (9.3 mg, 37%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.94(s,1H),9.63(s,1H),9.56(s,1H),8.90(dd,J=2.9,6.5Hz,1H),8.44(d,J=8.9Hz,1H),8.35( s,1H),8.26(d,J=8.6Hz,1H),8.13-8.05(m,2H),7.53-7.45(m,4H),6.74(s,1H),4.63(d,J=5.1Hz,1H),4.61-4.48(m,3 H), 4.42-4.33(m,3H), 3.95-3.77(m,4H), 2.89-2.82(m,1H), 2.81-2.71(m,1H), 2.59-2.48(m,5H), 2.40-2.29(m,2H), 2.23(dd,J=7.7,13.3Hz,1H), 2.08(ddd,J=4.5,9.1,13.3Hz,1H), 1.76-1.64(m,4H), 1.06-1.01(m,9H). LRMS(ESI) calculated values: C 49 H 53 35 ClN7O6S[M+H] + =902.3; Measured value: 902.3.

[0398] Synthesized target compound 20: 4-chloro-11-(1-(7-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-7-oxoheptanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL100)

[0399]

[0400] Following the synthetic method of LJL058, LJK056 (13 mg, 0.03 mmol), the known compound LJL098 (20 mg, 0.03 mmol), HATU (15 mg, 0.04 mmol), and DIEA (18 mg, 0.14 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL100 (10.8 mg, 33%), was obtained by HPLC purification and was a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.98-9.85(m,1H),9.60(s,1H),9.14(s,1H),8.88(dd,J=3.2,6.1Hz,1H),8.42(dd,J=2.3,9.3Hz,1H),8.32(s ,1H),8.23(d,J=9.0Hz,1H),8.12-8.03(m,2H),7.49-7.44(m,2H),7.43-7.38(m,2H),6.74(s,1H),4.64(d,J=4.9Hz,1H),4.60-4.49 (m,3H), 4.42-4.31(m,3H), 3.94-3.77(m,4H), 2.88-2.80(m,1H), 2.79-2.69(m,1H), 2.57-2.52(m,1H), 2.51-2.46(m,4H), 2.35-2.26(m,2H), 2.25-2.18(m,1H), 2.08(ddd,J=3.9,9.3,13.0Hz,1H), 1.73-1.62(m,4H), 1.48-1.38(m,2H), 1.03(s,9H). LRMS(ESI) calculated values: C 50 H 55 35 ClN7O6S[M+H] + =916.4; Measured value: 916.4.

[0401] Synthesized target compound 21: 4-chloro-11-(1-(8-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-8-oxooctanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL101)

[0402]

[0403] Following the synthetic method of LJL058, LJK056 (14 mg, 0.04 mmol), known compound LJL099 (23 mg, 0.04 mmol), HATU (16 mg, 0.04 mmol), and DIEA (20 mg, 0.16 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL101 (13.3 mg, 36%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.93(s,1H),9.65-9.57(m,1H),9.02(s,1H),8.89(dd,J=2.8,6.6Hz,1H),8.43(d,J=8.8Hz,1H),8.3 4(s,1H),8.29-8.22(m,1H),8.12-8.04(m,2H),7.48-7.39(m,4H),6.74(s,1H),4.64(d,J=5.1Hz,1H),4.60-4.47(m,3H), 4.41-4.32 (m, 3H), 3.94-3.76 (m, 4H), 2.89-2.81 (m, 1H), 2.80-2.71 (m, 1H), 2.56-2.44 (m, 5H), 2.35-2.26 (m, 2H), 2.24-2.18 (m, 1H), 2.08 (ddd, J = 4.5, 9.1, 13.2 Hz, 1H), 1.71-1.59 (m, 4H), 1.46-1.35 (m, 4H), 1.05-1.00 (m, 9H). LRMS (ESI) calculated value: C 51 H 57 35 ClN7O6S[M+H] + =930.4; Measured value: 930.4.

[0404] Synthesized target compound 22: 4-chloro-11-(1-(9-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-9-oxonanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL069)

[0405]

[0406] Following the synthetic method of LJL058, LJK056 (10 mg, 0.03 mmol), known compound LJL066 (17 mg, 0.03 mmol), HATU (12 mg, 0.03 mmol), and DIEA (14 mg, 0.11 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL069 (6 mg, 22%), was obtained by HPLC purification and was a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.94(s,1H),9.79(s,1H),9.63(s,1H),8.90(dd,J=2.7,6.6Hz,1H),8.44(d,J=9.0Hz,1 H),8.36(s,1H),8.26(d,J=8.9Hz,1H),8.12-8.04(m,2H),7.56-7.47(m,4H),6.74(s,1H),4.64-4.48(m,4H), 4.45-4.28 (m, 3H), 3.96-3.75 (m, 4H), 2.89-2.81 (m, 1H), 2.81-2.71 (m, 1H), 2.59-2.44 (m, 5H), 2.31-2.19 (m, 3H), 2.07 (ddd, J = 4.4, 9.3, 13.3 Hz, 1H), 1.70-1.57 (m, 4H), 1.43-1.31 (m, 6H), 1.02 (s, 9H). LRMS (ESI) calculated values: C 52 H 59 35 ClN7O6S[M+H] + =944.4; Measured value: 944.4.

[0407] Synthesized target compound 23: 4-chloro-11-(1-(2-(2-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-2-oxoethoxy)acetyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL096)

[0408]

[0409] Following the synthesis method of LJL058, LJK056 (12 mg, 0.03 mmol), the known compound LJL094 (18 mg, 0.03 mmol), HATU (14 mg, 0.04 mmol), and DIEA (17 mg, 0.13 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL096 (8.1 mg, 27%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.94-9.83(m,1H),9.62-9.46(m,1H),9.27-9.13(m,1H),8.91-8.80(m,1H),8.43-8.36( m,1H),8.28(d,J=9.4Hz,1H),8.24-8.15(m,1H),8.10-8.03(m,2H),7.49-7.34(m,4H),6.71(s,1H),4.71(d,J =8.3Hz, 1H), 4.63-4.45(m, 5H), 4.40-4.28(m, 3H), 4.24-4.10(m, 2H), 3.98-3.87(m, 2H), 3.86-3.75(m, 2H), 2.88-2.71(m, 2H), 2.51-2.43(m, 3H), 2.30-2.20(m, 1H), 2.14-2.05(m, 1H), 1.09-1.03(m, 9H). LRMS(ESI) calculated value: C 47 H 49 35 ClN7O7S[M+H] + =890.3; Measured value: 890.3.

[0410] Synthesized target compound 24: 4-chloro-11-(1-(4-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-4-oxobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL122)

[0411]

[0412] LJL118 (10.7 mg, 0.02 mmol) and lithium hydroxide monohydrate (5 mg, 0.12 mmol) were dissolved in tetrahydrofuran / methanol / water (volume ratio 2:1:3) and stirred at 50 °C for 1.5 hours. After the reaction was complete, the pH was adjusted to approximately 6 with 1 N hydrochloric acid solution, the solvent was removed under reduced pressure to obtain a solid, which was washed with diethyl ether and dried to obtain crude LJL119. LJL119, SM4 (8 mg, 0.02 mmol), HATU (8 mg, 0.02 mmol), and DIEA (10 mg, 0.08 mmol) were dissolved in DMF and stirred at room temperature for 0.5 hours. After the reaction was complete, the system was quenched with water, extracted with a dichloromethane / methanol mixed solvent, the organic phase was concentrated under reduced pressure, and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL122 (4.7 mg, 17%), as a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 10.02 (s, ¹H), 9.70 (s, ¹H), 9.29 (s, ¹H), 8.93 (dd, J = 2.3, 7.1 Hz, ¹H), 8.55–8.43 (m, 2H), 8.29 (dd, J = 1.7, 9.0 Hz, 1H), 8.15–8.07 (m, 2H), 7.55–7.38 (m, 4H). 6.72 (s, 1H), 4.68-4.17 (m, 6H), 4.05-3.73 (m, 4H), 3.50-3.42 (m, 1H), 3.41-3.33 (m, 2H), 3.22-3.07 (m, 2H), 2.65-2.43 (m, 5H), 2.25-2.04 (m, 4H), 1.06 (s, 9H). LRMS (ESI) calculated value: C 47 H 51 35 ClN7O5S[M+H] + =860.3; Measured value: 860.3.

[0413] Synthesized target compound 25: 4-chloro-11-(1-(5-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-5-oxopentyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL116)

[0414]

[0415] LJK056 (12 mg, 0.03 mmol), the known compound LJL115 (20 mg, 0.03 mmol), potassium carbonate (9 mg, 0.07 mmol), and potassium iodide (11 mg, 0.07 mmol) were dissolved in acetonitrile / DMF (1:1, v / v) and reacted at 60 °C for 4 hours. After the reaction was completed, the solution was concentrated under reduced pressure and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL116 (7.5 mg, 25%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.92(s,1H),9.59(s,1H),8.94(s,1H),8.87(dd,J=2.1,7.3Hz,1H),8.50-8.32(m,2H),8.17(dd,J =1.6,9.0Hz,1H),8.12-8.04(m,2H),7.47-7.35(m,4H),6.67(s,1H),4.67-4.48(m,4H),4.37(d,J=15.6Hz,1H),4.31-4. 17(m,1H), 4.05-3.75(m,4H), 3.55-3.40(m,1H), 3.37-3.32(m,2H), 3.21-3.02(m,2H), 2.52-2.36(m,5H), 2.26(dd,J=7.7,13.3Hz,1H), 2.10(ddd,J=4.4,9.2,13.3Hz,1H), 1.94-1.84(m,2H), 1.82-1.69(m,2H), 1.07(s,9H). LRMS(ESI) calculated value: C 48 H 53 35 ClN7O5S[M+H] + =874.4; Measured value: 874.4.

[0416] Synthesized target compound 26: 4-chloro-11-(1-(6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-6-oxohexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL082)

[0417]

[0418] Following the synthesis method of LJL116, using LJK056 (11 mg, 0.03 mmol), the known compound LJL081 (19 mg, 0.03 mmol), potassium carbonate (9 mg, 0.06 mmol), and potassium iodide (10 mg, 0.06 mmol) as raw materials, the trifluoroacetate of the target compound LJL082 (5.7 mg, 20%) was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.86(s,1H),9.52(s,1H),8.93(s,1H),8.83(dd,J=2.0,7.4Hz,1H),8.38(d,J=9.0Hz,1H),8.32(s,1H), 8.17-8.01(m,3H),7.48-7.37(m,4H),6.64(s,1H),4.67-4.48(m,4H),4.40-4.19(m,2H),4.06-3.85(m,3H),3.81(dd,J=3.8, 10.9Hz, 1H), 3.52-3.40(m, 1H), 3.36-3.32(m, 2H), 3.21-3.05(m, 2H), 2.46(s, 3H), 2.37(t, J = 7.5Hz, 2H), 2.28-2.20(m, 1H), 2.09(ddd, J = 4.4, 9.1, 13.3Hz, 1H), 1.95-1.84(m, 2H), 1.74(p, J = 7.4Hz, 2H), 1.55-1.43(m, 2H), 1.05(s, 9H). LRMS(ESI) calculated values: C 49 H 55 35 ClN7O5S[M+H] + =888.4; Measured value: 888.4.

[0419] Synthesized target compound 27: 4-chloro-11-(1-(7-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-7-oxoheptyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL104)

[0420]

[0421] Following the synthesis method of LJL116, using LJK056 (15 mg, 0.04 mmol), the known compound LJL102 (26 mg, 0.04 mmol), potassium carbonate (9 mg, 0.06 mmol), and potassium iodide (10 mg, 0.06 mmol) as raw materials, the trifluoroacetate of the target compound LJL104 (7 mg, 18%) was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.83(s,1H),9.48(s,1H),8.92(s,1H),8.81(dd,J=1.8,7.7Hz,1H),8.36(d,J=9.0Hz,1H),8.29(s,1H),8.1 4-8.02(m,3H),7.47-7.35(m,4H),6.62(s,1H),4.70-4.46(m,4H),4.42-4.20(m,2H),4.06-3.86(m,3H),3.82(dd,J=3.8,10.9Hz, 1H), 3.52-3.41(m,1H), 3.37-3.32(m,2H), 3.20-3.03(m,2H), 2.45(s,3H), 2.33(td,J=2.4,7.2Hz,2H), 2.25(dd,J=7.7,13.3Hz,1H), 2.10(ddd,J=4.5,9.2,13.3Hz,1H), 1.94-1.84(m,2H), 1.69(p,J=7.1Hz,2H), 1.53-1.41(m,4H), 1.05(s,9H). LRMS(ESI) calculated values: C 50 H 57 35 ClN7O5S[M+H] + =902.4; Measured value: 902.4.

[0422] Synthesized target compound 28: 4-chloro-11-(1-(8-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-8-oxooctyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL105)

[0423]

[0424] Following the synthesis method of LJL116, LJK056 (15 mg, 0.04 mmol), the known compound LJL103 (26 mg, 0.04 mmol), potassium carbonate (9 mg, 0.06 mmol), and potassium iodide (10 mg, 0.06 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL105 (9 mg, 23%), was obtained by HPLC purification and was a yellow solid. 1 H NMR (400MHz, methanol-d4) δ10.00(s,1H),9.68(s,1H),9.10(s,1H),8.92(dd,J=2.2,7.2Hz,1H),8.50(d,J=9.0Hz,1H),8.45(s,1H), 8.27(dd,J=1.7,9.1Hz,1H),8.15-8.06(m,2H),7.52-7.38(m,4H),6.71(s,1H),4.69-4.43(m,4H),4.41-4.22(m,2H),4.06-3. 86 (m, 3H), 3.82 (dd, J = 3.8, 11.0 Hz, 1H), 3.52-3.40 (m, 1H), 3.36-3.32 (m, 2H), 3.19-3.08 (m, 2H), 2.49 (s, 3H), 2.37-2.19 (m, 3H), 2.09 (ddd, J = 4.4, 9.2, 13.3 Hz, 1H), 1.92-1.80 (m, 2H), 1.71-1.59 (m, 2H), 1.50-1.35 (m, 6H), 1.04 (s, 9H). LRMS (ESI) calculated value: C 51 H 59 35 ClN7O5S[M+H] + =916.4; Measured value: 916.4.

[0425] Synthesized target compound 29: 4-chloro-11-(1-(9-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-9-oxonyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL114)

[0426]

[0427] Following the synthesis method of LJL116, using LJK056 (14 mg, 0.04 mmol), the known compound LJL113 (25 mg, 0.04 mmol), potassium carbonate (11 mg, 0.08 mmol), and potassium iodide (13 mg, 0.08 mmol) as raw materials, the trifluoroacetate of the target compound LJL114 (9.9 mg, 27%) was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.88(s,1H),9.53(s,1H),8.96(s,1H),8.84(dd,J=2.0,7.4Hz,1H),8.40(d,J=9.0Hz,1H),8.33(s,1H) ,8.15(d,J=8.8Hz,1H),8.11-8.02(m,2H),7.48-7.38(m,4H),6.64(s,1H),4.67-4.49(m,4H),4.41-4.21(m,2H),4.07-3.85( m, 3H), 3.81 (dd, J = 3.8, 11.0 Hz, 1H), 3.52-3.39 (m, 1H), 3.37-3.32 (m, 2H), 3.21-3.04 (m, 2H), 2.47 (s, 3H), 2.35-2.20 (m, 3H), 2.09 (ddd, J = 4.5, 9.1, 13.3 Hz, 1H), 1.94-1.83 (m, 2H), 1.68-1.59 (m, 2H), 1.49-1.34 (m, 8H), 1.04 (s, 9H). LRMS (ESI) calculated values: C 52 H 61 35 ClN7O5S[M+H] + =930.4; Measured value: 930.4.

[0428] Synthesized target compound 30: 4-bromo-11-(4-(9-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazo-5-yl)benzyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-9-oxonyl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL143)

[0429]

[0430] Following the synthetic method of LJL116, using 3-11 (16 mg, 0.04 mmol), the known compound LJL113 (25 mg, 0.04 mmol), potassium carbonate (11 mg, 0.08 mmol), and potassium iodide (13 mg, 0.08 mmol) as raw materials, the trifluoroacetate of the target compound LJL143 (3.7 mg, 9%) was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.62(s,1H),9.18(s,1H),8.91(s,1H),8.82(dd,J=1.0,8.4Hz,1H),8.36-8.26(m,2H),7 .95-7.86(m,2H),7.57(d,J=2.4Hz,1H),7.49-7.38(m,4H),4.70-4.44(m,4H),4.36(d,J=15.5Hz,1H),3.91(d ,J=11.0Hz,1H), 3.81(dd,J=3.9,11.0Hz,1H), 3.28-3.22(m,2H), 2.47(s,3H), 2.37-2.18(m,3H), 2.09(ddd,J=4.5,9.2,13.3Hz,1H), 1.88-1.77(m,2H), 1.67-1.58(m,2H), 1.49-1.33(m,8H), 1.04(s,9H). LRMS(ESI) calculated value: C 51 H 62 79 BrN8O5S[M+H] + =977.4; Measured value: 977.4.

[0431] Synthesized target compound 31: 4-chloro-11-(1-(9-(((S)-1-((2S,4R)-4-hydroxy-2-(((S)-1-(4-(4-methylthiazo-5-yl)phenyl)ethyl)carbamoyl)pyrrolidine-1-yl)-3,3-dimethyl-1-oxobutane-2-yl)amino)-9-oxonyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL147)

[0432]

[0433] Following the synthetic method of LJL116, using LJK056 (17 mg, 0.05 mmol), the known compound LJL145 (39 mg, 0.06 mmol), potassium carbonate (14 mg, 0.10 mmol), and potassium iodide (16 mg, 0.10 mmol) as raw materials, the trifluoroacetate 4-15 of the target compound was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.92(s,1H),9.59(s,1H),8.97(s,1H),8.87(dd,J=1.9,7.4Hz,1H),8.43(d,J=9.0Hz,1H),8.38(s,1H),8.19(dd,J =1.6,8.9Hz,1H),8.12-8.04(m,2H),7.47-7.39(m,4H),6.67(s,1H),5.01(q,J=7.0Hz,1H),4.67-4.54(m,2H),4.48-4.40(m,1H),4.32-4 .20(m,1H), 4.03-3.85(m,3H), 3.76(dd,J=3.9,11.0Hz,1H), 3.50-3.40(m,1H), 3.37-3.32(m,2H), 3.20-3.06(m,2H), 2.48(s,3H), 2.35-2.17(m,3H), 1.96(ddd,J=4.5,9.0,13.3Hz,1H), 1.92-1.83(m,2H), 1.69-1.60(m,2H), 1.52-1.36(m,11H), 1.05(s,9H). LRMS(ESI) calculated value: C 53 H 63 35 ClN7O5S[M+H] + =944.4; Measured value: 944.4.

[0434] Synthesized target compound 32: 4-chloro-11-(1-((S)-5-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-5-(4-(4-methylthiazolyl-5-yl)phenyl)pentyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJK061)

[0435]

[0436] LJK056 (12 mg, 0.03 mmol), the known compound LJK060 (19 mg, 0.03 mmol), NaBH(OAc)3 (10 mg, 0.05 mmol), and triethylamine (5 mg, 0.05 mmol) were dissolved in dichloromethane / methanol (3:1, v / v) and stirred at room temperature for 12 hours. After the reaction was complete, the system was quenched with water, extracted with a dichloromethane / methanol mixture, concentrated under reduced pressure, and purified by HPLC to obtain the trifluoroacetate salt of the target compound, LJK061 (9.3 mg, 30%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.96(s,1H),9.62(s,1H),9.06(s,1H),8.89(dd,J=2.4,7.0Hz,1H),8.45(d,J=9.0Hz,1H),8.40(s,1H),8.22(dd,J=1.6,9.0 Hz,1H),8.15-8.04(m,2H),7.50-7.46(m,4H),6.69(s,1H),5.03-4.98(m ,1H),4.77(d,J=9.0Hz,1H),4.69-4.57(m,1H),4.50-4.42(m,1H),4.37- 4.22(m, 1H), 4.09-3.90(m, 2H), 3.87(d, J = 11.1 Hz, 1H), 3.78(dd, J = 3.6, 11.1 Hz, 1H), 3.54-3.43(m, 1H), 3.42-3.32(m, 2H), 3.24-3.04(m, 2H), 2.51(s, 3H), 2.22(dd, J = 7.5, 13.1 Hz, 1H), 2.03-1.86(m, 5H), 1.79-1.66(m, 1H), 1.64-1.52(m, 1H), 1.39-1.26(m, 4H), 1.09(s, 9H). LRMS(ESI) calculated value: C 51 H 56 35 Cl 19FN7O5S[M+H] + =932.4; Measured value: 932.4.

[0437] Synthesized target compound 33: 4-chloro-11-(1-((S)-6-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJM112)

[0438]

[0439] Following the synthetic method of LJK061, LJK056 (120 mg, 0.33 mmol), the known compound LJM111 (200 mg, 0.33 mmol), NaBH(OAc)3 (106 mg, 0.50 mmol), and triethylamine (50 mg, 0.50 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJM112 (239 mg, 75%), was obtained by HPLC purification and was a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.84(s,1H),9.47(s,1H),8.98(s,1H),8.81(dd,J=2.3,7.1Hz,1H),8.35(d,J=8.9Hz,1H),8.29(s,1H),8.13-8. 01(m,3H),7.48-7.43(m,4H),6.62(s,1H),4.98-4.92(m,1H),4.77(dd,J=1.2,9.4Hz,1H),4.70-4.57(m,1H),4.46(s,1H),4.34-4.17( m,1H),4.08-3.88(m,2H),3.86(d,J=11.2Hz,1H),3.78(dd,J=3.6,11.1Hz,1H),3.52-3.41(m,1H),3.40-3.33(m,2H),3.23-3.01(m,2H ),2.49(s,3H),2.21(dd,J=7.6,13.1Hz,1H),1.99-1.85(m,5H),1.74-1.63(m,1H),1.61-1.50(m,3H),1.39-1.27(m,4H),1.09(s,9H). 13 C NMR(101MHz,MeOD)δ173.45,171.57,171.55,171.33(d,J C-F=20.4Hz),163.09,154.10,145.27,144.79,141.80,137.32,136.51,135.73,135.15,134.47,133.72,131 .83,130.42,130.32,129.60,129.45,128.28,127.49,124.33,123.73,123.13,122.26,117.80,79.20(d,J C-F =232.6Hz),70.95,60.67,58.76,58.68,58.14,57.33,54.26,51.84,50 .10,38.82,37.29,36.97,27.13,26.92,26.70,24.97,14.88,14.06(d,J C-F =10.2Hz), 13.89(d,J) C-F =10.4Hz). LRMS (ESI) calculated value: C 52 H 58 35 Cl 19 FN7O5S[M+H] + =946.4; Measured value: 946.4.

[0440] Synthesized target compound 34: 4-chloro-11-(1-((S)-7-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-7-(4-(4-methylthiazolyl-5-yl)phenyl)heptyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJM150)

[0441]

[0442] Following the synthetic method of LJK061, LJK056 (16 mg, 0.05 mmol), the known compound LJM149 (28 mg, 0.05 mmol), NaBH(OAc)3 (14 mg, 0.07 mmol), and triethylamine (7 mg, 0.07 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJM150 (12.6 mg, 28%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.76(s,1H),9.39(s,1H),8.94(s,1H),8.76(dd,J=2.1,7.3Hz,1H),8.29(d,J=9.0Hz,1H),8.21(s,1H),8.07-7.99 (m,3H),7.47-7.41(m,4H),6.57(s,1H),4.95-4.91(m,1H),4.76(d,J=8.3Hz,1H),4.64(t,J=8.5Hz,1H),4.49-4.42(m,1H),4.33-4.16(m ,1H), 4.08-3.88(m,2H), 3.85(d,J=11.1Hz,1H), 3.77(dd,J=3.7,11.1Hz,1H), 3.52-3.39(m,1H), 3.38-3.32(m,2H), 3.23-3.00(m,2H), 2.48(s,3H), 2.25-2.16(m,1H), 1.99-1.83(m,5H), 1.67-1.57(m,1H), 1.57-1.46(m,5H), 1.37-1.27(m,4H), 1.08(s,9H). LRMS(ESI) calculated values: C 53 H 60 35 Cl 19 FN7O5S[M+H] + =960.4; Measured value: 960.4.

[0443] Synthesized target compound 35: 4-chloro-11-(1-((S)-8-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-8-(4-(4-methylthiazolyl-5-yl)phenyl)octyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJN025)

[0444]

[0445] Following the synthetic method of LJK061, LJK056 (13 mg, 0.04 mmol), the known compound LJN024 (23 mg, 0.04 mmol), NaBH(OAc)3 (12 mg, 0.05 mmol), and triethylamine (6 mg, 0.05 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJN025 (32 mg, 89%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.88(s,1H),9.55(s,1H),9.09(s,1H),8.84(dd,J=3.4,5.9Hz,1H),8.40(d,J=9.0Hz,1H),8.34(s,1H),8.16(dd,J=1.5,9.0H z,1H),8.09-8.01(m,2H),7.49-7.41(m,4H),6.66(s,1H),4.92-4.89(m, 1H),4.76(dd,J=1.2,9.4Hz,1H),4.63(dd,J=7.6,9.1Hz,1H),4.49-4.42( m, 1H), 4.37-4.22(m, 1H), 4.04-3.87(m, 2H), 3.84(d, J = 11.2 Hz, 1H), 3.76(dd, J = 3.7, 11.1 Hz, 1H), 3.51-3.39(m, 1H), 3.37-3.32(m, 2H), 3.21-3.01(m, 2H), 2.51(s, 3H), 2.24-2.16(m, 1H), 1.99-1.80(m, 5H), 1.63-1.53(m, 1H), 1.51-1.41(m, 7H), 1.37-1.25(m, 4H), 1.08(s, 9H). LRMS(ESI) calculated values: C 54 H 62 35 Cl 19 FN7O5S[M+H] + = 974.4; Measured value: 974.4.

[0446] Synthesized target compound 36: 4-bromo-11-(4-((S)-5-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-5-(4-(4-methylthiazolyl-5-yl)phenyl)pentyl)piperazin-1-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJK045)

[0447]

[0448] Following the synthetic method of LJK061, LJK050 (19 mg, 0.05 mmol), the known compound LJK060 (27 mg, 0.05 mmol), NaBH(OAc)3 (15 mg, 0.07 mmol), and triethylamine (7 mg, 0.07 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJK045 (3.3 mg, 7%), was obtained by HPLC purification and was a yellow solid.1 H NMR (400MHz, methanol-d4) δ9.64(s,1H),9.20(s,1H),8.95(s,1H),8.86-8.81(m,1H),8.37-8.26(m,2H),7.95- 7.88(m,2H),7.59(d,J=2.4Hz,1H),7.49-7.45(m,4H),5.01-4.95(m,1H),4.76(d,J=8.9Hz,1H),4.63-4. 58 (m, 1H), 4.47-4.44 (m, 1H), 3.86 (d, J = 11.3 Hz, 1H), 3.79-3.75 (m, 1H), 2.49 (s, 3H), 2.25-2.18 (m, 1H), 1.99-1.87 (m, 5H), 1.74-1.64 (m, 1H), 1.60-1.53 ​​(m, 1H), 1.41-1.25 (m, 6H), 1.08 (s, 9H). LRMS (ESI) calculated value: C 50 H 57 79 Br 19 FN8O5S[M+H] + =779.3; Measured value: 779.3.

[0449] Synthesized target compound 37: 4-bromo-11-(4-((S)-6-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)piperazin-1-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJM133)

[0450]

[0451] Following the synthetic method of LJK061, LJK050 (272 mg, 0.67 mmol), the known compound LJM111 (400 mg, 0.67 mmol), NaBH(OAc)3 (212 mg, 1.0 mmol), and triethylamine (101 mg, 1.0 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJM133 (445 mg, 67%), was obtained by HPLC purification and was a yellow solid. 1¹H NMR (400MHz, methanol-d⁴) δ 9.52 (s, 1H), 9.05 (s, 1H), 8.94 (s, 1H), 8.73 (d, J = 8.2 Hz, 1H), 8.27–8.20 (m, 2H), 7.90–7.80 (m, 2H), 7.49–7.43 (m, 5H), 4.95–4.92 (m, 1H), 4.75 (dd, J = 1.2, 9.2 Hz, 1H), 4.62 (dd, J = 7.6, 9.2 Hz, 1H) ),4.47-4.43(m,1H),3.85(d,J=11.0Hz,1H),3.77(dd,J=3.7,11.0Hz,1H),3.30-3.23(m,2H),2.49(s,3H), 2.25-2.16(m,1H),1.99-1.83(m,5H),1.70-1.59(m,1H),1.56-1.48(m,3H),1.38-1.27(m,4H),1.07(s,9H). 13 C NMR(101MHz,MeOD)δ173.42,171.54,171.52,171.26(d,J C-F =20.4Hz), 161.75(TFA,q,J C-F =36.9Hz),160.29,154.41,154.13,146.46,145.30,141.17,140.90,138.47,135.73,134.73,132.65 ,131.40,131.21,130.38,130.17,128.27,124.47,123.51,121.92,121.75,117.91,117.49(TFA,q,J C-F =291.9Hz), 106.09, 79.18 (d, J) C-F =232.7Hz),70.92,60.64,58.73,58.65,58.10,57.98,54.26,52.23,45 .06,38.80,37.27,36.93,27.06,26.90,26.65,24.62,14.81,14.06(d,J C-F =10.5Hz), 13.88(d,J) C-F =10.6Hz). LRMS (ESI) calculated value: C 51 H 59 79 Br 19 FN8O5S[M+H] + =993.3; Measured value: 993.3.

[0452] Synthesized target compound 38: 4-chloro-11-(1-((6S)-6-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-3-methyl-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL142)

[0453]

[0454] Step 1: Synthesis of compound 11-(1-((6S)-6-((tert-butoxycarbonyl)amino)-3-methyl-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-4-chloro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-compound (LJL140)

[0455] LJK056 (37 mg, 0.10 mmol), the known compound LJL139 (50 mg, 0.10 mmol), potassium carbonate (29 mg, 0.21 mmol), and potassium iodide (17 mg, 0.10 mmol) were dissolved in acetonitrile / DMF (3:1, v / v) and reacted at 80 °C for 4 hours. After the reaction was completed, the solution was concentrated under reduced pressure and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL140 (29.3 mg, 37%), as a yellow liquid. 1 H NMR (400MHz, methanol-d4) δ9.95 (s, 1H), 9.63 (s, 1H), 9.18 (s, 1H), 8.89 (dd, J = 3.2, 6.2Hz, 1H), 8.46 (d, J = 8. 9Hz,1H),8.41(s,1H),8.23(dd,J=1.6,9.0Hz,1H),8.13-8.04(m,2H),7.51-7.43(m,4H),6.69(s,1H), 4.63-4.51 (m, 1H), 4.36-4.19 (m, 1H), 4.01-3.87 (m, 2H), 3.51-3.33 (m, 3H), 3.25-3.08 (m, 2H), 2.52 (s, 3H), 1.95-1.76 (m, 3H), 1.74-1.61 (m, 2H), 1.47-1.26 (m, 11H), 1.02 (t, J = 6.5 Hz, 3H). LRMS (ESI) calculated value: C 43 H 47 35 ClN5O3S[M+H] +=748.3; Measured value: 748.3.

[0456] Step 2: Synthesize compound 4-chloro-11-(1-((6S)-6-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-3-methyl-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJL142)

[0457] LJL140 (29 mg, 0.04 mmol) was dissolved in methanol, and hydrochloric acid (0.4 mmol, 4 M in Dioxane) was added. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the solvent was removed under reduced pressure, and the crude product was washed with diethyl ether and dried. The crude product, known compound LJN001 (13 mg, 0.04 mmol), HATU (16 mg, 0.04 mmol), and DIEA (20 mg, 0.15 mmol) were dissolved in DMF and stirred at room temperature for 0.5 hours. After the reaction was complete, the system was quenched with water, extracted with a dichloromethane / methanol mixed solvent, and the organic phase was concentrated under reduced pressure. The purified product was then purified by HPLC to obtain the trifluoroacetate of the target compound, LJL142 (9.5 mg, 25%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.92(s,1H),9.58(s,1H),8.98(s,1H),8.87(d,J=6.9Hz,1H),8.42(d,J=8.6Hz,1H),8.36(s,1H),8.18(d,J= 8.7Hz,1H),8.13-8.03(m,2H),7.49-7.44(m,4H),6.66(s,1H),4.95-4.92(m,1H),4.76(d,J=9.0Hz,1H),4.70-4.59(m,1H),4.46(s ,1H), 4.35-4.20(m,1H), 4.05-3.89(m,2H), 3.86(d,J=11.0Hz,1H), 3.78(dd,J=3.3,10.9Hz,1H), 3.53-3.34(m,3H), 3.23-3.04(m,2H), 2.49(s,3H), 2.26-2.16(m,1H), 1.99-1.83(m,4H), 1.80-1.64(m,3H), 1.41-1.30(m,5H), 1.13-1.01(m,12H). LRMS(ESI) calculated values: C 53 H 60 35 Cl19 FN7O5S[M+H] + =960.4; Measured value: 960.4.

[0458] Synthesized target compound 39: 4-chloro-11-(1-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)pent-4-yn-1-yl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG032)

[0459]

[0460] Following the synthetic method of LJK061, LJK056 (18 mg, 0.05 mmol), the known compound LJK160 (21 mg, 0.06 mmol), NaBH(OAc)3 (21 mg, 0.10 mmol), and triethylamine (10 mg, 0.10 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG032 (8.6 mg, 25%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, DMSO-d6) δ11.03(s,1H),9.51(s,1H),9.28(s,1H),8.59(dd,J=3.5,5.9Hz,1H),8.14(d,J=9.0H z,1H),7.99(s,1H),7.93-7.87(m,3H),7.74(d,J=7.6Hz,1H),7.69(d,J=7.4Hz,1H),7.55(t,J=7.6Hz,1H),6. 53 (s, 1H), 5.18 (dd, J = 5.1, 13.3 Hz, 1H), 4.53 (d, J = 17.7 Hz, 1H), 4.37 (d, J = 17.7 Hz, 1H), 4.24-4.09 (m, 1H), 4.06-3.81 (m, 2H), 3.01-2.88 (m, 3H), 2.78-2.55 (m, 4H), 2.47-2.35 (m, 2H), 2.21-1.96 (m, 4H). LRMS (ESI) calculated values: C 39 H 33 35 ClN5O4[M+H] + =670.2; Measured value: 670.2.

[0461] Synthesized target compound 40: 4-chloro-11-(1-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hex-5-yn-1-yl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK151)

[0462]

[0463] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJK145 (14 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJK151 (4.5 mg, 15%), was obtained by HPLC purification and was a yellow solid. 1 H NMR (400MHz, methanol-d4) δ10.01(s,1H),9.68(s,1H),8.93(dd,J=2.3,7.1Hz,1H),8.50(d,J=9.0Hz,1H),8.42(s,1H),8.26(dd,J=1.7,9.0Hz,1H), 8.15-8.08(m,2H),7.70(d,J=7.6Hz,1H),7.63(dd,J=1.1,7.7Hz,1H),7.48(t,J=7.7Hz,1H),6.72(s,1H),5.18(dd,J=5.2,13.3Hz,1H),4.58-4 .44(m,2H), 4.36-4.24(m,1H), 4.07-3.86(m,2H), 3.56-3.47(m,1H), 3.42(dd,J=5.9,10.4Hz,2H), 3.21-3.07(m,2H), 2.98-2.87(m,1H), 2.84-2.76(m,1H), 2.65(t,J=6.8Hz,2H), 2.52(qd,J=4.6,13.2Hz,1H), 2.24-2.15(m,1H), 2.13-2.03(m,2H), 1.80(p,J=7.1Hz,2H). LRMS(ESI) calculated value: C 40 H 35 35 ClN5O4[M+H] + =684.2; Measured value: 684.2.

[0464] Synthesized target compound 41: 4-chloro-11-(1-(7-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hept-6-yn-1-yl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG027)

[0465]

[0466] Following the synthetic method of LJK061, using LJK056 (14 mg, 0.04 mmol), the known compound LJK152 (14 mg, 0.04 mmol), NaBH(OAc)3 (20 mg, 0.09 mmol), and triethylamine (10 mg, 0.09 mmol) as raw materials, the trifluoroacetate of the target compound LCG027 (9.1 mg, 33%) was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.79(s,1H),9.43(s,1H),8.80(dd,J=2.0,7.3Hz,1H),8.32(d,J=8.9Hz,1H),8.22(s,1H),8.10-8.01(m, 3H),7.67(dd,J=1.0,7.7Hz,1H),7.60-7.55(m,1H),7.44(t,J=7.6Hz,1H),6.60(s,1H),5.17(dd,J=5.2,13.3Hz,1H),4.61-4.3 8 (m, 2H), 4.38-4.17 (m, 1H), 4.05-3.83 (m, 2H), 3.51-3.42 (m, 1H), 3.40-3.34 (m, 2H), 3.20-3.01 (m, 2H), 2.97-2.87 (m, 1H), 2.83-2.75 (m, 1H), 2.61-2.47 (m, 3H), 2.24-2.15 (m, 1H), 2.02-1.91 (m, 2H), 1.81-1.73 (m, 2H), 1.73-1.64 (m, 2H). LRMS (ESI) calculated value: C 41 H 37 35 ClN5O4[M+H] + =698.3; Measured value: 698.3.

[0467] Synthesized target compound 42: 4-chloro-11-(1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oct-7-yn-1-yl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG031)

[0468]

[0469] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJK156 (20 mg, 0.05 mmol), NaBH(OAc)3 (23 mg, 0.11 mmol), and triethylamine (11 mg, 0.11 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG031 (2.9 mg, 9%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.81(s,1H),9.45(s,1H),8.81(d,J=7.4Hz,1H),8.35(d,J=9.0Hz,1H),8.26(s,1H),8.16-7.97(m,3H ),7.72(dd,J=1.1,7.7Hz,1H),7.61(dd,J=1.1,7.6Hz,1H),7.49(t,J=7.6Hz,1H),6.60(s,1H),5.18(dd,J=5.1,13.3Hz,1H) ,4.63-4.39(m,2H),4.33-4.18(m,1H),4.06-3.83(m,2H),3.51-3.40(m,1H),3.39-3.33(m,2H),3.22-3.03(m,2H),2.97-2.87(m,1H),2.84-2.75(m,1H),2.60-2.45(m,3H),2.24-2.15(m,1H),2.01-1.87(m,2H),1.78-1.52(m,6H). LRMS(ESI) calculated value: C 42 H 39 35 ClN5O4[M+H] + =712.3; Measured value: 712.3.

[0470] Synthesized target compound 43: 4-chloro-11-(1-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)propyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL025)

[0471]

[0472] Following the synthetic method of LJK061, LJK056 (16 mg, 0.05 mmol), the known compound LJL015 (14 mg, 0.05 mmol), NaBH(OAc)3 (20 mg, 0.09 mmol), and triethylamine (9 mg, 0.09 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL025 (4 mg, 13%), was obtained by HPLC purification and was a yellow solid. 1 ¹H NMR (400MHz, methanol-d⁴) δ 9.79 (s, 1H), 9.47 (s, 1H), 8.90–8.77 (m, 1H), 8.48–8.25 (m, 2H), 8.18–7.86 (m, 3H), 7.72 (d, J = 7.2 Hz, 1H), 7.65–7.46 (m, 2H), 6.61 (s, 1H), 5.22 (dd, J = 5.1, 13.4 Hz, 1H) ,4.67-4.43(m,2H),4.35-4.16(m,1H),4.16-3.86(m,2H),3.44-3.36(m,1H),3.28-3.19(m,2H),3.19-3.02(m,2H),3.01-2.67(m,4H),2.60-2.46(m,1H),2.38-2.16(m,3H). LRMS(ESI) calculated values: C 37 H 33 35 ClN5O4[M+H] + =646.2; Measured value: 646.3.

[0473] Synthesized target compound 44: 4-chloro-11-(1-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)butyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL024)

[0474]

[0475] Following the synthetic method of LJK061, using LJK056 (15 mg, 0.04 mmol), the known compound LJL009 (13 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) as raw materials, the trifluoroacetate of the target compound LJL024 (12.2 mg, 44%) was purified by HPLC and obtained as a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.72(s,1H),9.33(s,1H),8.74(d,J=7.7Hz,1H),8.27(d,J=8.9Hz,1H),8.17(s,1H),8.11-7. 88(m,3H),7.68(dd,J=1.3,7.3Hz,1H),7.58-7.47(m,2H),6.53(s,1H),5.19(dd,J=5.2,13.3Hz,1H),4.66-4.46(m, 2H), 4.32-4.15(m,1H), 4.06-3.79(m,2H), 3.53-3.41(m,1H), 3.41-3.34(m,2H), 3.24-3.14(m,1H), 3.11-2.99(m,1H), 2.96-2.81(m,3H), 2.80-2.70(m,1H), 2.58-2.45(m,1H), 2.24-2.15(m,1H), 2.03-1.80(m,4H). LRMS(ESI) calculated value: C 38 H 35 35 ClN5O4[M+H] + =660.2; Measured value: 660.2.

[0476] Synthesized target compound 45: 4-chloro-11-(1-(5-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)pentyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL023)

[0477]

[0478] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJL002 (14 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL023 (7.7 mg, 27%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.82(s,1H),9.47(s,1H),8.81(d,J=7.4Hz,1H),8.36(d,J=8.5Hz,1H),8.28(s,1H),8.23-7.91(m,3 H),7.67(dd,J=1.6,7.0Hz,1H),7.54-7.48(m,2H),6.61(s,1H),5.20(dd,J=5.1,13.3Hz,1H),4.59-4.47(m,2H),4.35-4. 10(m,1H), 4.02-3.81(m,2H), 3.46-3.40(m,1H), 3.37-3.33(m,2H), 3.23-3.02(m,2H), 2.98-2.88(m,1H), 2.84-2.75(m,3H), 2.59-2.48(m,1H), 2.24-2.16(m,1H), 1.97-1.87(m,2H), 1.83(q,J=7.6Hz,2H), 1.55(q,J=7.7Hz,2H). LRMS(ESI) calculated value: C 39 H 37 35 ClN5O4[M+H] + =674.3; Measured value: 674.4.

[0479] Synthesized target compound 46: 4-chloro-11-(1-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG026)

[0480]

[0481] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJK147 (14 mg, 0.04 mmol), NaBH(OAc)3 (17 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG026 (6.5 mg, 22%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.76(s,1H),9.37(s,1H),8.76(d,J=7.6Hz,1H),8.29(d,J=9.1Hz,1H),8.20(s,1H),8.13-7.94(m,3H),7. 65(dd,J=1.9,6.7Hz,1H),7.54-7.42(m,2H),6.55(s,1H),5.18(dd,J=5.1,13.3Hz,1H),4.61-4.43(m,2H),4.31-4.17(m,1H),4 0.02-3.82(m,2H), 3.51-3.37(m,1H), 3.30-3.26(m,2H), 3.21-3.12(m,1H), 3.10-3.00(m,1H), 2.97-2.86(m,1H), 2.84-2.70(m,3H), 2.53(qd,J=4.6,13.3Hz,1H), 2.24-2.15(m,1H), 1.97-1.83(m,2H), 1.82-1.71(m,2H), 1.58-1.44(m,4H). LRMS(ESI) calculated value: C 40 H 39 35 ClN5O4[M+H] + =688.3; Measured value: 688.3.

[0482] Synthesized target compound 47: 4-chloro-11-(1-(7-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)heptyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG028)

[0483]

[0484] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJK154 (15 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG028 (5.2 mg, 17%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.40(s,1H),8.97(s,1H),8.55(d,J=7.6Hz,1H),8.08-7.96(m,2H),7.97-7.86(m,2H),7.69(d,J= 9.1Hz,1H),7.63(dd,J=1.4,7.2Hz,1H),7.53-7.41(m,2H),6.37(s,1H),5.18(dd,J=5.2,13.3Hz,1H),4.61-4.41(m,2H ), 4.27-4.15(m,1H), 4.00-3.81(m,2H), 3.46-3.37(m,1H), 3.36-3.32(m,2H), 3.09-2.84(m,3H), 2.84-2.68(m,3H), 2.53(qd,J=4.6,13.2Hz,1H), 2.23-2.15(m,1H), 1.99-1.85(m,2H), 1.82-1.71(m,2H), 1.63-1.37(m,6H). LRMS(ESI) calculated values: C 41 H 41 35 ClN5O4[M+H] + =702.3; Measured value: 702.3.

[0485] Synthesized target compound 48: 4-chloro-11-(1-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)octyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG029)

[0486]

[0487] Following the synthetic method of LJK061, LJK056 (15 mg, 0.04 mmol), the known compound LJK158 (16 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG029 (9.5 mg, 31%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.62(s,1H),9.19(s,1H),8.71-8.64(m,1H),8.18(d,J=8.9Hz,1H),8.10-7.97(m,3H),7.86(d,J=9.1Hz,1H ),7.63(dd,J=2.1,6.6Hz,1H),7.50-7.42(m,2H),6.47(s,1H),5.17(dd,J=5.1,13.3Hz,1H),4.58-4.41(m,2H),4.30-4.15(m,1H ), 4.02-3.80(m,2H), 3.50-3.38(m,1H), 3.36-3.32(m,2H), 3.24-3.12(m,1H), 3.09-2.97(m,1H), 2.97-2.86(m,1H), 2.84-2.69(m,3H), 2.53(qd,J=4.7,13.2Hz,1H), 2.23-2.15(m,1H), 1.97-1.84(m,2H), 1.77-1.67(m,2H), 1.54-1.36(m,8H). LRMS(ESI) calculated values: C 42 H 43 35 ClN5O4[M+H] + =716.3; Measured value: 716.2.

[0488] Synthesized target compound 49: 4-bromo-11-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hex-5-yn-1-yl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJK136)

[0489]

[0490] Following the synthetic method of LJK061, LJK050 (17 mg, 0.04 mmol), the known compound LJK145 (14 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJK136 (2.5 mg, 8%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.59 (s, 1H), 9.14 (s, 1H), 8.80 (dd, J = 1.0, 8.4Hz, 1H), 8.34-8.23 (m, 2H), 7.95-7. 83(m,2H),7.73(dd,J=1.1,7.7Hz,1H),7.61(dd,J=1.0,7.7Hz,1H),7.54-7.47(m,2H),5.19(dd,J=5.1, 13.3Hz, 1H), 4.55-4.46(m, 2H), 3.37-3.33(m, 2H), 2.98-2.88(m, 1H), 2.83-2.76(m, 1H), 2.63(t, J = 6.8Hz, 2H), 2.58-2.47(m, 1H), 2.23-2.16(m, 1H), 2.10-2.00(m, 2H), 1.78(p, J = 7.0Hz, 2H). LRMS(ESI) calculated value: C 39 H 36 79 BrN6O4[M+H] + =731.2; Measured value: 731.3.

[0491] Synthesized target compound 50: 4-chloro-11-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hex-5-yn-1-yl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG016)

[0492]

[0493] Following the synthetic method of LJK061, LCG013 (15 mg, 0.04 mmol), the known compound LJK145 (14 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG016 (5.2 mg, 18%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.64(s,1H),9.20(s,1H),8.79(dd,J=2.1,7.3Hz,1H),8.34(d,J=9.5Hz,1H),8.09-7.99(m,2H) ,7.90(dd,J=2.4,9.5Hz,1H),7.74(d,J=7.3Hz,1H),7.63(dd,J=1.0,7.7Hz,1H),7.57(d,J=2.3Hz,1H),7.50(t,J=7. 6Hz, 1H), 5.20 (dd, J = 5.2, 13.3Hz, 1H), 4.57-4.45 (m, 2H), 3.38-3.33 (m, 2H), 2.97-2.87 (m, 1H), 2.83-2.75 (m, 1H), 2.64 (t, J = 6.8Hz, 2H), 2.58-2.47 (m, 1H), 2.23-2.16 (m, 1H), 2.10-1.99 (m, 2H), 1.78 (p, J = 7.2Hz, 2H). LRMS (ESI) calculated value: C 39 H 36 35 ClN6O4[M+H] + =687.2; Measured value: 687.2.

[0494] Synthesized target compound 51: 4-bromo-11-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hexyl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG025)

[0495]

[0496] Following the synthetic method of LJK061, LJK050 (13 mg, 0.03 mmol), the known compound LJK147 (11 mg, 0.03 mmol), NaBH(OAc)3 (14 mg, 0.06 mmol), and triethylamine (6 mg, 0.06 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG025 (11.6 mg, 49%), was obtained by HPLC purification and was a yellow solid. 1¹H NMR (400MHz, methanol-d⁴) δ 9.55 (s, ¹H), 9.09 (s, ¹H), 8.77 (dd, J = 1.1, 8.4Hz, ¹H), 8.30–8.21 (m, 2H), 7.92–7.80 (m, 2H), 7.63 (dd, J = 2.6, 6.1Hz, ¹H), 7.53–7.42 (m, 3H), 5.17 (dd, J = 5.1, 13.3Hz) ,1H), 4.55-4.43(m,2H), 3.26-3.19(m,2H), 2.96-2.86(m,1H), 2.83-2.69(m,3H), 2.53(qd,J=4.7,13.2Hz,1H), 2.24-2.15(m,1H), 1.86-1.69(m,4H), 1.51-1.42(m,4H). LRMS(ESI) calculated values: C 39 H 40 79 BrN6O4[M+H] + =735.2; Measured value: 735.2.

[0497] Synthesized target compound 52: 4-chloro-11-(4-(6-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)hexyl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG023)

[0498]

[0499] Following the synthetic method described in 4-16, LCG013 (15 mg, 0.04 mmol), the known compound LJK147 (14 mg, 0.04 mmol), NaBH(OAc)3 (18 mg, 0.08 mmol), and triethylamine (8 mg, 0.08 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG023 (14.6 mg, 51%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.54(s,1H),9.08(s,1H),8.71(dd,J=2.7,6.7Hz,1H),8.26(d,J=9.4Hz,1H),8.02- 7.95(m,2H),7.84(dd,J=2.3,9.5Hz,1H),7.62(dd,J=2.5,6.1Hz,1H),7.51-7.42(m,3H),5.16(dd,J=5.2, 13.3Hz, 1H), 4.54-4.42(m, 2H), 3.25-3.19(m, 2H), 2.97-2.85(m, 1H), 2.83-2.69(m, 3H), 2.53(qd, J=4.7, 13.2Hz, 1H), 2.22-2.15(m, 1H), 1.88-1.77(m, 2H), 1.76-1.67(m, 2H), 1.51-1.42(m, 4H). LRMS(ESI) calculated value: C 39 H 40 35 ClN6O4[M+H] + =691.3; Measured value: 691.3.

[0500] Synthesized target compound 53: 4-bromo-11-(4-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)octyl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LCG030)

[0501]

[0502] Following the synthetic method of LJK061, LJK050 (13 mg, 0.03 mmol), the known compound LJK158 (13 mg, 0.03 mmol), NaBH(OAc)3 (14 mg, 0.07 mmol), and triethylamine (7 mg, 0.07 mmol) were used as raw materials. The trifluoroacetate of the target compound, LCG030 (9.8 mg, 40%), was obtained by HPLC purification and was a yellow solid. 1¹H NMR (400MHz, methanol-d⁴) δ 9.52 (s, ¹H), 9.07 (s, ¹H), 8.75 (dd, J = 1.1, 8.4 Hz, ¹H), 8.29–8.21 (m, 2H), 7.91–7.79 (m, 2H), 7.66–7.60 (m, ¹H), 7.52–7.42 (m, 3H), 5.16 (dd, J = 5.2, 13.3 Hz, ¹H), 4.54–4. 42 (m, 2H), 3.26-3.18 (m, 2H), 2.96-2.85 (m, 1H), 2.82-2.68 (m, 3H), 2.53 (qd, J = 4.7, 13.2 Hz, 1H), 2.23-2.14 (m, 1H), 1.85-1.75 (m, 2H), 1.75-1.65 (m, 2H), 1.46-1.34 (m, 8H). LRMS (ESI) calculated values: C 41 H 44 79 BrN6O4[M+H] + =763.3; Measured value: 763.3.

[0503] Synthesized target compound 54: 4-chloro-11-(4-(8-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)octyl)piperazin-1-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL003)

[0504]

[0505] Following the synthetic method of LJK061, LCG013 (14 mg, 0.04 mmol), the known compound LJK158 (14 mg, 0.04 mmol), NaBH(OAc)3 (16 mg, 0.06 mmol), and triethylamine (6 mg, 0.06 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL003 (13.4 mg, 48%), was purified by HPLC and obtained as a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.54(s,1H),9.09(s,1H),8.71(dd,J=2.2,7.3Hz,1H),8.26(d,J=9.4Hz,1H),8.04-7.94(m,2H),7.8 4(dd,J=2.4,9.5Hz,1H),7.66-7.60(m,1H),7.53-7.42(m,3H),5.16(dd,J=5.2,13.3Hz,1H),4.55-4.42(m,2H),3.25-3.1 8(m,2H), 2.90(ddd,J=5.3,13.4,17.7Hz,1H), 2.78(ddd,J=2.4,4.7,17.7Hz,1H), 2.71(t,J=7.6Hz,2H), 2.53(qd,J=4.7,13.2Hz,1H), 2.18(dtd,J=2.5,5.3,12.9Hz,1H), 1.84-1.74(m,2H), 1.73-1.65(m,2H), 1.44-1.34(m,8H). LRMS(ESI) calculated value: C 41 H 44 35 ClN6O4[M+H] + =719.3; Measured value: 719.3.

[0506] Synthesized target compound 55: 4-chloro-11-(1-(6-((2-(2,6-dioxoperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)hexanoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL037)

[0507]

[0508] LJK056 (23 mg, 0.06 mmol), the known compound LJL036 (25 mg, 0.06 mmol), HATU (27 mg, 0.07 mmol), and DIEA (33 mg, 0.26 mmol) were dissolved in DMF and stirred at room temperature for 1 hour. After the reaction was complete, the system was diluted with water, extracted with a dichloromethane / methanol mixed solvent, concentrated under reduced pressure, and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL037 (16 mg, 33%), as a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.96-9.89(m,1H),9.62-9.56(m,1H),8.92-8.84(m,1H),8.44-8.36(m,1H),8.30(s,1H),8 .24(t,J=8.9Hz,1H),8.14-8.06(m,2H),7.50-7.38(m,1H),7.02(d,J=8.6Hz,1H),6.93-6.87(m,1H),6.78-6.72( m, 1H), 5.03 (dd, J=5.5, 12.5Hz, 1H), 4.44-4.33 (m, 2H), 3.95-3.83 (m, 2H), 3.38-3.33 (m, 1H), 3.28-3.24 (m, 1H), 2.85-2.64 (m, 5H), 2.60-2.50 (m, 2H), 2.13-2.05 (m, 1H), 1.79-1.64 (m, 4H), 1.58-1.46 (m, 2H). LRMS(ESI) calculated values: C 40 H 36 35 ClN6O6[M+H] + =731.2; Measured value: 731.2.

[0509] Synthesized target compound 56: 4-chloro-11-(1-(3-(2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)ethoxy)ethoxy)propionyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL040)

[0510]

[0511] Following the synthetic method of LJL037, LJK056 (15 mg, 0.04 mmol), known compound LJL039 (18 mg, 0.04 mmol), HATU (17 mg, 0.05 mmol), and DIEA (21 mg, 0.17 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL040 (17 mg, 52%), was obtained by HPLC purification and was a yellow solid. 1¹H NMR (500MHz, methanol-d⁴) δ 9.89–9.84 (m, 1H), 9.49 (s, 1H), 8.85–8.78 (m, 1H), 8.36–8.31 (m, 1H), 8.26–8.16 (m, 2H), 8.12–8.05 (m, 2H), 7.30–7.20 (m, 1H), 6.83–6.56 (m, 3H), 5.07–5.00 (m, 1H), 4.52–4.28 (m, 2H), 3.92–3.79 (m, 4H), 3.65–3.55 (m, 4H), 3.30–3.21 (m, 3H), 2.88–2.63 (m, 8H), 2.13–2.05 (m, 1H). LRMS (ESI) calculated values: C 41 H 38 35 ClN6O8[M+H] + =777.2; Measured value: 777.2.

[0512] Synthesized target compound 57: 4-chloro-11-(1-(6-((2-(2,6-dioxoperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL043)

[0513]

[0514] Following the synthetic method of LJK061, LJK056 (19 mg, 0.05 mmol), the known compound LJL042 (20 mg, 0.05 mmol), NaBH(OAc)3 (23 mg, 0.11 mmol), and triethylamine (11 mg, 0.11 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL043 (16.2 mg, 41%), was purified by HPLC and obtained as a yellow solid. 1H NMR (400MHz, methanol-d4) δ9.81(s,1H),9.44(s,1H),8.79(d,J=7.0Hz,1H),8.39-8.23(m,2H),8.15-7.97(m,3H),7.53( dd,J=7.0,8.6Hz,1H),7.09-6.96(m,2H),6.61(s,1H),5.06(dd,J=5.5,12.5Hz,1H),4.39-4.11(m,1H),4.07-3.77 (m,2H), 3.52-3.40(m,1H), 3.40-3.32(m,4H), 3.23-2.97(m,2H), 2.87(ddd,J=5.1,13.9,17.3Hz,1H), 2.80-2.62(m,2H), 2.12(tdd,J=2.9,5.9,8.2Hz,1H), 1.98-1.84(m,2H), 1.79-1.67(m,2H), 1.64-1.46(m,4H). LRMS(ESI) calculated values: C 40 H 38 35 ClN6O5[M+H] + =717.3; Measured value: 717.3.

[0515] Synthesized target compound 58: 4-chloro-11-(1-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperazin-1-yl)benzoyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL044)

[0516]

[0517] Following the synthetic method of LJL037, LJK056 (15 mg, 0.04 mmol), the known compound LJL044-S (19 mg, 0.04 mmol), HATU (17 mg, 0.05 mmol), and DIEA (21 mg, 0.17 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL044 (16.2 mg, 48%), was purified by HPLC and obtained as a yellow solid. 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.74(s,1H),9.55(s,1H),8.80-8.69(m,1H),8.33(d,J=8.9Hz,1H),8.23-8 .08(m,2H),7.96(d,J=4.5Hz,2H),7.72(d,J=8.5Hz,1H),7.53-7.36(m,3H),7.32(dd,J=2.3,8.6Hz,1H),7.05(d, J = 8.4 Hz, 2H), 6.70 (s, 1H), 5.08 (dd, J = 5.4, 12.9 Hz, 1H), 4.34 (s, 2H), 3.85-3.76 (m, 2H), 3.65 (t, J = 5.3 Hz, 4H), 3.45 (t, J = 5.3 Hz, 4H), 2.94-2.84 (m, 1H), 2.79-2.68 (m, 2H), 2.65-2.53 (m, 2H), 2.07-1.98 (m, 1H). LRMS (ESI) calculated values: C 45 H 37 35 ClN7O6[M+H] + =806.2; Measured value: 806.2.

[0518] Synthesized target compound 59: 4-chloro-11-(1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL027)

[0519]

[0520] Following the synthetic method of LJK061, LJK056 (25 mg, 0.07 mmol), the known compound LJL026 (25 mg, 0.07 mmol), NaBH(OAc)3 (29 mg, 0.14 mmol), and triethylamine (14 mg, 0.14 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJL027 (4.6 mg, 9%), was obtained by HPLC purification and was a yellow solid. 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.56(s,1H),9.34(s,1H),8.62(dd,J=2.8,6.6Hz,1H),8.18(d,J=9.0Hz,1H) ,8.04(s,1H),7.98-7.88(m,3H),7.69(d,J=8.5Hz,1H),7.39(d,J=2.2Hz,1H),7.30(dd,J=2.3,8.7Hz,1H),6.53(s, 1H), 5.08 (dd, J = 5.4, 12.8 Hz, 1H), 4.27-4.08 (m, 3H), 4.01-3.82 (m, 2H), 3.22 (d, J = 6.7 Hz, 2H), 3.06 (t, J = 12.4 Hz, 2H), 2.99-2.79 (m, 3H), 2.65-2.51 (m, 3H), 2.36-2.25 (m, 1H), 2.08-1.89 (m, 3H), 1.44-1.27 (m, 2H). LRMS (ESI) calculated values: C 40 H 36 35 ClN6O5[M+H] + =715.2; Measured value: 715.2.

[0521] Synthesized target compound 60: 4-chloro-11-(1-((1-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)methyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL084)

[0522]

[0523] Step 1: Synthesize compound 11-(1-((1-(tert-butoxycarbonyl)piperidin-4-yl)methyl)-1,2,3,6-tetrahydropyridin-4-yl)-4-chloro-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL080)

[0524] LJK056 (20 mg, 0.06 mmol), 1-Boc-4-piperidinecarboxaldehyde (18 mg, 0.08 mmol), NaBH(OAc)3 (23 mg, 0.11 mmol), and triethylamine (11 mg, 0.11 mmol) were dissolved in dichloromethane / methanol (3:1, v / v) and stirred at room temperature for 12 hours. After the reaction was complete, the system was quenched with water, extracted with a dichloromethane / methanol mixture, concentrated under reduced pressure, and purified by HPLC to obtain the trifluoroacetate of the target compound, LJL080 (13.3 mg, 42%), as a yellow solid. 1 ¹H NMR (400 MHz, methanol-d⁴) δ 9.81 (s, ¹H), 9.45 (s, ¹H), 8.79 (dd, J = 2.4, 6.9 Hz, ¹H), 8.33 (d, J = 9.0 Hz, ¹H), 8.27 (s, ¹H), 8.08–8.01 (m, ³H), 6.58 (s, ¹H), 4.38–3.86 (m, ⁵H), 3.6 3-3.40 (m, 1H), 3.26 (d, J = 7.0 Hz, 2H), 3.24-2.98 (m, 2H), 2.96-2.78 (m, 2H), 2.30-2.17 (m, 1H), 1.93 (d, J = 13.0 Hz, 2H), 1.47 (s, 9H), 1.32-1.26 (m, 4H). LRMS (ESI) calculated values: C 32 H 36 35 ClN4O3[M+H] + =559.2; Measured value: 559.2.

[0525] Step 2: Synthesize compound 4-chloro-11-(1-((1-((1-(2-(2,6-dioxopiridin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperidin-4-yl)methyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJL084)

[0526] LJL080 (13 mg, 0.02 mmol) was dissolved in methanol, and hydrochloric acid (0.2 mmol, 4 M in dioxane) was added. The mixture was stirred at room temperature for 12 hours. After the reaction was complete, the solvent was removed under reduced pressure. The crude product was washed with diethyl ether, dried, and used directly in the next reaction. The above crude product, known compound LJL026 (9 mg, 0.02 mmol), sodium cyanoborohydride (7 mg, 0.03 mmol), and triethylamine (4 mg, 0.03 mmol) were dissolved in dichloromethane / methanol (3:1, v / v) and stirred at room temperature for 12 hours. After the reaction was complete, the system was quenched with water, extracted with a dichloromethane / methanol mixture, and the organic phase was concentrated under reduced pressure. The purified product by HPLC yielded the trifluoroacetate of the target compound, LJL084 (9 mg, 47%), as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.73(s,1H),9.34(s,1H),8.74(dd,J=1.6,7.8Hz,1H),8.27(d,J=9.0Hz,1H),8.17(s,1H),8.11-7.88(m, 3H),7.63(d,J=8.5Hz,1H),7.31(d,J=2.3Hz,1H),7.20(dd,J=2.3,8.7Hz,1H),6.54(s,1H),5.07(dd,J=5.5,12.4Hz,1H),4.36- 3.99(m,4H), 3.86-3.65(m,3H), 3.53-3.33(m,3H), 3.27-2.93(m,8H), 2.92-2.81(m,1H), 2.80-2.62(m,2H), 2.52-2.39(m,1H), 2.37-2.16(m,3H), 2.15-2.06(m,1H), 1.95(d,J=12.4Hz,2H), 1.79(q,J=11.4Hz,2H), 1.43(q,J=11.5Hz,2H). LRMS(ESI) calculated value: C 46 H 47 35 ClN7O5[M+H] + =812.3; Measured value: 812.5.

[0527] Synthesized target compound 61: 4-chloro-11-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)amino)heptyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinoline[3,2-a]phthalazine-7-onthium-6-oxide (LJN026)

[0528]

[0529] Following the synthetic method of LJK061, LJK056 (33 mg, 0.09 mmol), the known compound LJN026-S (35 mg, 0.09 mmol), NaBH(OAc)3 (29 mg, 0.14 mmol), and triethylamine (14 mg, 0.14 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJN026 (19 mg, 28%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.88(s,1H),9.53(s,1H),8.83(dd,J=2.0,7.3Hz,1H),8.40(d,J=8.9Hz,1H),8.32(s,1H),8.15(dd,J=1.6,8.9H z,1H),8.10-8.03(m,2H),7.51(d,J=8.4Hz,1H),6.94(d,J=2.1Hz,1H),6.81(dd,J=2.2,8.4Hz,1H),6.64(s,1H),5.02(dd,J=5.5,12.5 Hz, 1H), 4.33-4.18(m, 1H), 4.03-3.83(m, 2H), 3.51-3.39(m, 1H), 3.37-3.32(m, 2H), 3.23(t, J = 6.9Hz, 2H), 3.18-3.02(m, 2H), 2.84(ddd, J = 5.1, 13.8, 17.4Hz, 1H), 2.76-2.62(m, 2H), 2.12-2.04(m, 1H), 1.93-1.84(m, 2H), 1.74-1.65(m, 2H), 1.51(s, 6H). LRMS(ESI) calculated value: C 41 H 40 35 ClN6O5[M+H] + =731.3; Measured value: 731.3.

[0530] Synthesized target compound 62: 4-chloro-10-(1-((S)-6-((2S,4R)-1-((S)-2-(1-fluorocyclopropane-1-carbamate)-3,3-dimethylbutyryl)-4-hydroxypyrrolidine-2-carbamate)-6-(4-(4-methylthiazolyl-5-yl)phenyl)hexyl)-1,2,3,6-tetrahydropyridin-4-yl)-5-oxo-5H-isoquinolino[3,2-a]phthalazine-7-onthium-6-oxide (LJN027)

[0531]

[0532] Following the synthetic method of LJK061, LJM027 (12 mg, 0.03 mmol), the known compound LJM111 (20 mg, 0.03 mmol), NaBH(OAc)3 (11 mg, 0.05 mmol), and triethylamine (5 mg, 0.05 mmol) were used as raw materials. The trifluoroacetate of the target compound, LJN027 (20.6 mg, 65%), was purified by HPLC and obtained as a yellow solid. 1 H NMR (400MHz, methanol-d4) δ9.94(s,1H),9.61(s,1H),9.04(s,1H),8.88(dd,J=2.9,6.4Hz,1H),8.46(s,1H),8.39-8.28(m,2H),8.10-8.03(m,2H),7. 49-7.44(m,4H),6.61(s,1H),4.97-4.94(m,1H),4.76(dd,J=1.2,9.4Hz ,1H),4.63(t,J=8.5Hz,1H),4.46(dt,J=2.3,4.4Hz,1H),4.32-4.17(m,1 H), 4.06-3.88(m,2H), 3.86(d,J=11.3Hz,1H), 3.77(dd,J=3.7,11.1Hz,1H), 3.51-3.40(m,1H), 3.39-3.32(m,2H), 3.20-3.01(m,2H), 2.50(s,3H), 2.21(ddt,J=1.8,7.7,13.3Hz,1H), 1.98-1.85(m,5H), 1.72-1.62(m,1H), 1.59-1.50(m,3H), 1.38-1.27(m,4H), 1.09(s,9H). LRMS(ESI) calculated values: C 52 H 58 35 Cl 19 FN7O5S[M+H] + =946.4; Measured value: 946.4.

[0533] By selecting appropriate raw materials, the compounds listed in Table 1 below can be synthesized:

[0534] Table 1: Representative Compounds

[0535]

[0536]

[0537]

[0538]

[0539]

[0540]

[0541]

[0542]

[0543]

[0544]

[0545]

[0546]

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559] Example 2: FP detection of the binding force constant K between the compound and the SMARCA2 bromine domain i value

[0560] His-tag-labeled SMARCA2 (1373-1511aa) protein was expressed in *E. coli*. The protein was first purified using a Ni affinity column and then further purified using a Superdex 75 molecular sieve, yielding a SMARCA2 protein purity greater than 95% and a concentration of 222.5 μM. Based on the purified SMARCA2 protein and the fluorescent molecular probe F1, the dissociation constant K between the fluorescent molecular probe F1 and SMARCA2 was determined using a fluorescence biased-positive assay. d It is 59.44 nM.

[0561] 96-well plates were purchased from Corning (black, #3694). The multi-plate reader was a TECAN SPARK 10M. The compound dilution buffer and FP detection buffer were the same: 10mM Tris (pH 7.5), 200mM NaCl (Sigma), 0.01% Tween-20, and 0.01% Trition X-100 (Sigma). The experimental water was Millipore-Q purified water.

[0562] First, the test compound was dissolved in DMSO to prepare a 10 mM standard stock solution. Then, the standard stock solution of the test compound was diluted in an EP tube with compound dilution buffer to prepare a working sample solution. The concentration of the prepared working sample solution was 50 times the highest sample concentration required on the test plate (50× test compound solution). The compound was serially diluted 3-fold in the EP tube for later use. 2 μL of the serially diluted 50× test compound solution of test compound A was added to wells B1-D1 to B12-D12 of a 96-well plate, and 2 μL of the serially diluted 50× test compound solution of test compound B was added to wells E1-G1 to E12-G12. Then, 98 μL of detection buffer containing 5.1 nM fluorescent molecular probe and 306 nM SMARCA2 protein was added to each well. Wells A1-A3 served as a blank control group: 100 μL of detection buffer was added. Wells A4-A6 served as a negative signal reference group: 100 μL of buffer containing only 5 nM fluorescent molecular probe F1 was added. Wells A7-A9 served as the positive control group: 100 μL of a mixed solution containing 5 nM fluorescent molecular probe F1 and 300 nM MARCA2 protein was added.

[0563] Cover the reaction plate with aluminum foil and incubate the 96-well plate on a 96-well plate shaker at room temperature for 1 hour. Then, read the fluorescence polarization mP values ​​at Ex 485 nm / Em 530 nm using a microplate reader. Plot the measured mP values ​​against the compound concentration gradient. The median of the mP maximum and minimum values ​​corresponds to the sample compound concentration, which is the IC50 of the compound binding to the protein. 50 value([I] 50 According to this IC 50value([I] 50 The binding rate between the compound and the protein was calculated using the formula Changshu K. i :K i =[I] 50 / ([L] 50 / K d +[P]0 / K d +1). Where [L] 50 [P]0 represents 50% of the concentration of the fluorescently labeled molecular probe in the above test system; [P]0 represents the concentration of SMARCA2 protein in the above test system, K d It is the dissociation constant of proteins and fluorescently labeled molecular probes.

[0564] Using the above method, the binding force constant K of the SMARCA2 bromine domain in some compounds and the literature compound ACBI2,AU-15330 was tested. i The values ​​and data are shown in Table 2.

[0565]

[0566] Table 2: Binding force constant K of the bromine domain of the compound SMARCA2 in the examples i value

[0567]

[0568]

[0569] ***1nM-100nM, **100nM-10μM, *10μM-100μM.

[0570] Experimental Conclusions: The experimental results show that the compounds have an affinity for binding to the SMARCA2 protein. Some compounds exhibit particularly strong affinity, with a binding force constant K... i In the nM interval.

[0571] Example 3: The inhibitory activity of the compound on the proliferation of SMARCA4 mutant cells NCI-H1944

[0572] NCI-H1944 cells were seeded in 96-well plates at a density of 1500 cells per well. The next day, serially diluted solutions were added and incubated for 5 days. Then, 100 μL of 10% TCA was added to each well and the cells were fixed at 4°C for 1 hour. The TCA was discarded, and the cells were rinsed three times with water and then dried in an oven or air-dried. Next, 100 μL of 0.4% (w / v) sulfonylrhodamine B (SRB) prepared with 1% acetic acid was added to each well. After rinsing several times with 1% (v / v) acetic acid, the cells were dried in an oven or air-dried. Finally, 200 μL of 10 mM Tris solution was added to each well, and the cells were vortexed in a microplate reader to mix. The OD value was read at 560 nm. Inhibition rate (%) = (OD...) Control -OD Compound ) / OD Control ×100%, the half-maximal inhibitory concentration (IC50) was calculated using a four-parameter model fitting based on the inhibition rate of each compound concentration. 50 )value.

[0573] Table 3: Cell proliferation inhibitory activity of compounds in the examples

[0574]

[0575]

[0576] ***1nM-100nM, **100nM-10μM, *10μM-100μM.

[0577] Experimental conclusion: The compound can inhibit the proliferation of SMARCA4 mutant NCI-H1944 cells and has anti-cancer activity.

[0578] Example 4: The inhibitory activity of the compound on the proliferation of SMARCA4 mutant cells NCI-H1568

[0579] Dissolve the test sample in 100% dimethyl sulfoxide (DMSO) to prepare a 10 mM stock solution of the compound. Dilute the compound with 100% DMSO to the highest concentration required for the experiment (1 mM or 5 mM). First, add 115 μL of complete cell culture medium to wells B1-G1 of a 96-well flat-bottomed clear cell culture plate, and add 80 μL of complete culture medium to wells B2-G12. Then, add 5 μL of the highest concentration of the compound solution required to wells B1-D1 and E1-G1, and then serially dilute 3-fold to wells B12-D12 and E12-G12. Finally, add 40 μL of NCI-H1568 cell solution to each well, with approximately 2500-3000 cells per well, for a total volume of 120 μL per well. In addition to the compound being tested, two control groups were set up: 1) a control group with cells and complete culture medium but no compound; 2) a control group with only complete culture medium, no cells, and no compound. After incubating 96-well plates in a 37°C cell culture incubator containing 5% CO2 for 5 days, 12 μL of CCK-8 reagent was added to each well, followed by incubation at 37°C for 2-4 hours. The absorbance of the 96-well plates was read using a TECAN microplate reader at 450 nm. The effect of different concentrations of the compound on cell viability was calculated using the following formula: [Experimental group absorbance - Absorbance of only complete culture medium (cell-free, compound-free group)] / [Absorbance of group with cells but no compound - Absorbance of only complete culture medium (cell-free, compound-free group)] × 100%. The data were processed using GraphPad Prism 10 software, and the IC50 values ​​were... 50 The value represents the concentration of the compound that inhibits cell growth by 50%.

[0580] Table 4: Cell proliferation inhibitory activity of compounds in the examples

[0581]

[0582]

[0583] ***1nM-100nM, **100nM-10μM, *10μM-100μM.

[0584] Experimental conclusion: The compound can inhibit the proliferation of SMARCA4 mutant NCI-H1568 cells and has anti-cancer activity.

[0585] Example 5: Activity of the compound in degrading SMARCA2 in NCI-H1944 cells

[0586] NCI-H1944 cells were seeded in 24-well plates and cultured at 37°C and 5% CO2. On the second day, the final concentration of the test compound (100 nM) was added to the wells. DMSO and the reported SMARCA2 degrader AU-15330 were used as solvent and positive controls, respectively, and the cells were cultured for another 2 hours. Cells were collected and washed twice with pre-chilled PBS. Freshly prepared 1×SDS lysis buffer (50 mM Tris-HCl pH 8.1, 1% SDS, 100 mM DTT, 0.01% bromophenol blue) was added, and the cell lysate was collected into EP tubes. The cells were incubated at 100°C for 15 minutes before further processing or stored at -20°C for later use. 10 μg of the lysis buffer was subjected to SDS-PAGE gel electrophoresis and blotted onto the NC membrane. The membrane was blocked with TBST containing 5% (w / v) skim milk powder for 1 hour at room temperature, and then washed four times with TBST buffer for 5 minutes each time. The membrane was incubated overnight at 4°C with either SMARCA2 antibody (1:1000) or Vinculin antibody (1:1000). The membrane was washed four times with TBST buffer for 5 minutes each time and incubated at room temperature for 1 hour with horseradish peroxidase-linked secondary antibody (1:2000). The membrane was washed four times with TBST for 5 minutes each time, and luminescence images were captured using the ChemiDoc Touch Imaging system (BioRad). The immunoblot was quantified using ImageLab software.

[0587] Table 5: Activity of Compounds in Examples in Degrading Intracellular SMARCA2

[0588]

[0589] ***1nM-100nM; **100nM-10μM; *10μM-100μM; nc, not detected.

[0590] Experimental conclusion: The compound has the activity of degrading SMARCA2 in NCI-H1944 cells.

[0591] Example 6: Western Blot detection of the degradation of SMARCA2, SMARCA4 and PBRM1 by compounds in NCI-H1975 cells

[0592] NCI-H1975 cells were trypsinized and evenly seeded into 6-well plates, then incubated in a 37°C cell culture incubator containing 5% CO2 for 24 hours. After complete adhesion, the original culture medium was removed, and fresh culture medium containing different concentrations of compound 37 and ACBI2 was added to each well of the 6-well plate, with DMSO as a blank control. The plates were then incubated at 37°C for 2 hours. After incubation, the 6-well plates were removed, the compound-containing culture medium was removed, and each well was washed once with 1 mL of pre-chilled 1X PBS. After removing the PBS, 100 μL of RIPA (Beyotime, P0013B) cell lysis buffer was added to each well, and cells were lysed on ice for 30 minutes. After centrifugation at 13,000 rpm at 4°C for 1 hour, the cell lysis supernatant was collected. The concentration of each protein sample was determined using a BCA protein concentration assay kit (Beyotime, P0011). 20 μg of protein was loaded into each well for SDS-PAGE, and then the protein was transferred to a nitrocellulose membrane. Nitrocellulose membranes were blocked with 1×TBST (20mM Tris, 150mM NaCl, pH 7.6, 0.1% Tween-20) solution containing 5% skim milk powder for 1 hour at room temperature, followed by washing with 1×TBST for 5×5 minutes each time. Then, primary antibodies against the corresponding proteins (SMARCA2, SMARCA4, PBRM1, and Tubulin) were added and incubated overnight at 4°C. The next day, the primary antibodies were removed, the membrane was washed with 1×TBST for 5×5 minutes, the corresponding secondary antibodies were added, and the membrane was incubated at room temperature for 1 hour, followed by washing with 1×TBST for 5×5 minutes each time. Finally, the bands of the corresponding proteins were detected and analyzed using an ECL luminescence assay kit (Thermo Scientific). The images were processed using Photoshop and PowerPoint to obtain the corresponding image data.

[0593] The results are as follows Figure 1 As shown, compounds 37 and ACBI2 possess triple activity in degrading SMARCA2, SMARCA4, and PBRM1 in NCI-H1975 cells. After three replicates, the DCs of 37 degrading SMARCA2, SMARCA4, and PBRM1 showed increased activity. 50 The concentrations of ACBI2 at different DC values ​​for the degradation of SMARCA2, SMARCA4, and PBRM1 were 3.5 ± 0.09 nM, 7.0 ± 0.48 nM, and 6.41 ± 3.43 nM, respectively. 50 The values ​​were 3.99±1.16 nM, 6.19±3.84 nM, and 0.86±0.21 nM, respectively. The activity of 37 in degrading PBRM1 was weaker than that of ACBI2, but it had a certain advantage in terms of degradation selectivity.

[0594] Example 7: Antitumor efficacy of the compound in a mouse model

[0595] NCI-H1944 cells in logarithmic growth phase were seeded into the right axilla of BALB / c nude mice for tumor tissue growth, passage, and preservation. Tumor tissue in logarithmic growth phase was cut into 40mm pieces. 3 Small pieces were re-inoculated into the right axilla of BALB / c nude mice for subsequent experiments. The volume of the tumor tissue was measured using calipers, and the tumor was allowed to grow to 150 mm. 3 Animals were randomly divided into a solvent control group and a drug administration group. The compound was administered three times weekly via tail vein injection or once daily via intraperitoneal injection. The solvent control group received an equal volume of solvent for three weeks. Tumor volume and mouse weight were measured twice weekly during this period, and tumor growth and weight change curves were plotted. At the end of the experiment, tumor tissue was weighed and a bar chart was plotted. The formula for calculating tumor volume (V) is as follows: V = 0.5 × a 2 ×b, where a is the width of the tumor tissue and b is the length of the tumor tissue. The relative tumor volume (RTV) is calculated based on the measured tumor volume data as follows: RTV = V t / V0, where V0 represents the tumor volume of the mouse at the time of grouping, V t This represents the mouse tumor volume obtained in each measurement. The formula for calculating the relative tumor proliferation rate (T / C) is as follows: T / C(%) = RTV treatment / RTV control ×100%, of which RTV treatment and RTV control The RTVs are for the drug administration group and the solvent group, respectively.

[0596] The results are as follows Figure 2 As shown, Figure 2 In the diagram, 112 represents compound 33, 133 represents compound 37, and AU-15330 is a positive control compound. Experimental results show that in a mouse xenograft model of H1944 cancer cells, compound 33 at a dose of 50 mg / kg and compound 37 at a dose of 25 mg / kg significantly inhibited tumor growth, demonstrating good antitumor efficacy.

[0597] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound, or its enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts thereof, wherein, The compound is shown in Formula I; In the formula, R1 is selected from the following group: H, halogens, and hydroxyl groups; R2 is the ligand portion of the E3 ubiquitin ligase, or R A L1 ; R A L1 Selected from the following group: H, halogen, cyano, optionally substituted C1-C4 alkyl, -OR A P -N(R3)R A P -N(R3)C(O)R A P -C(O)R A P -C(O)OR A P -OC(O)R A P -C(O)N(R3)R A P Optionally substituted C3-C10 carbocyclic groups, Optionally substituted 3- to 10-membered heterocyclic groups, Optionally substituted C6-C10 aryl groups, Optionally substituted 5- to 10-membered heteroaryl groups, Optionally substituted -C3-C10 carbocyclic groups -N(R3)R A P -Optionally substituted -3 to 10-membered heterocyclic groups -N(R3)R A P -Optionally substituted -C6-C10 arylene-N(R3)R A P -Optionally substituted -5 to 10-membered heteroaryl-N(R3)R A P -Optionally substituted -C3-C10 subcarbonyl cyclogroup-OR A P -Optionally substituted -3 to 10-membered heterocyclic group -OR A P -Optionally substituted -C6-C10 arylene-OR A P and - optional substitution of -5 to 10-membered heteroaryl-OR A P ; R A P Selected from the following groups: H, C1-C4 alkyl, amino protecting group, hydroxy protecting group; R3 is hydrogen or a C1-C4 alkyl group; L represents none (single bond) or a connector; "Optionally substituted" means that the group is unsubstituted or that one or more hydrogens in the group are replaced by R. S Replaced; each R S Independently selected from the following group: halogen, -OR a -OC(O)R a -NR a R b -SR a -R a -CN, -NO2, -CO2R a -CONR a R b -C(O)R a -OC(O)NR a R b -NR b C(O)R a -NR b C(O)2R a -NR a -C(O)NR b R c -S(O)R a -S(O)2R a -S(O)2NR a R b -NR a S(O)2R b and -C1-C2 alkylene-R a ;where R a R b and R c Each is independently selected from the following group: hydrogen, deuterium, C1-C4 alkyl, C1-C4 haloalkyl, C1-C4 hydroxyalkyl, C1-C4 deuteralkyl, C1-C4 aminoalkyl, C3-C6 cycloalkyl, C2-C4 alkenyl, C2-C4 alkynyl, phenyl, 5-6 heteroaryl, C3-C6 cycloalkyl, 4 to 6 heterocyclic alkyl and benzyl.

2. The compound according to claim 1, characterized in that, R2 is the E3 ubiquitin ligase ligand moiety, and the E3 ubiquitin ligase ligand moiety is selected from the following group: In each formula, X1 is selected from the group consisting of: hydrogen, halogen, C1-C6 alkoxy, cyano (CN), optionally substituted C1-C6 alkyl; and / or, L is selected from the following group: Among the various types, m, n, m1, m2, n1, and n2 each independently represent 1, 2, 3, or 4 (preferably, m+n≤5, m1+n1≤5, and m2+n2≤5); R3 is hydrogen or a C1-C4 alkyl group; L1 is a divalent linker.

3. The compound according to claim 1, characterized in that, L1 is selected from the following group: optionally substituted C1-C12 alkylene, optionally substituted C1-C12 alkenyl, optionally substituted C1-C12 ynylene; Furthermore, the -CH2- group in the alkylene, alkenylene, and yntyl groups is independently surrounded by 0, 1, 2, 3, or 4 M groups. a and 0 or 1 M b What it replaces; among them, M a Each person independently selects from the following groups: -O-、-CO-、-NR7-、-CONR7-、-R7NCO-、-NHCONH-、-S-、-SO-、SO2-、 M b Independently selected from the following subcyclic groups: optionally substituted C3-C10 carbon rings, optionally substituted C6-C10 aromatic rings, optionally substituted 3- to 10-membered heterocycles and optionally substituted 5- to 10-membered heteroaromatic rings; The condition for attachment is: when there are two -CH2- particles being M a When substitution occurs, the two substituted -CH2- groups are separated by at least one C atom (preferably a C atom located on an alkylene chain, an alkenylene chain, or an alkynylene chain), and when both CH2 groups are substituted by groups selected from -O-, -NR7-, and -S-, the two substituted CH2 groups are separated by at least two C atoms (preferably C atoms located on an alkylene chain, an alkenylene chain, or an alkynylene chain). Each R7 is independently hydrogen or C1-C3 alkyl.

4. The compound according to claim 1, characterized in that, The compound is shown in Formula II; In the formula, R1, R2, and L are defined as in formula I of claim 1; Alternatively, the compound may be represented as shown in Formula III: In the formula, R1, R2 and L are defined as in Formula I of claim 1.

5. The compound according to claim 1, characterized in that, The compound is shown in Formula IV; In the formula, R2 and L are as defined in Formula I of claim 1; Alternatively, the compound may be represented as shown in Formula V; In the formula, R2 and L are as defined in Formula I of claim 1.

6. The compound according to claim 2, characterized in that, L1 is selected from the following group:

7. The compound according to claim 1, characterized in that, The compounds are selected from Table A.

8. A pharmaceutical composition comprising (i) a compound as claimed in any one of claims 1 to 7, or an enantiomer, diastereomer, racemate, or pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

9. The use of the compound as claimed in any one of claims 1 to 7, or its enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts, or the use of the pharmaceutical composition as claimed in claim 8, wherein... Used to prepare (i) inhibitors with SMARCA2 protein binding ability; and / or (ii) medicines for the prevention or treatment of diseases related to the activity or expression level of SMARCA2 and SMARCA4 proteins.

10. The use as described in claim 9, characterized in that, The diseases mentioned are selected from the following group: cancer, fibrotic diseases.

11. The use as described in claim 10, characterized in that, The cancers are selected from the following group: lung cancer, meningioma, mesothelioma, fibrosarcoma, bladder cancer, ovarian cancer, adenocarcinoma, gastric cancer, pancreatic cancer, prostate cancer, colon cancer, bone cancer, brain cancer, neurocytoma, rectal cancer, colon cancer, familial adenomatous polyposis, hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, kidney cancer, renal parenchymal carcinoma, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, testicular cancer, urinary tract cancer, melanoma, acute lymphoblastic leukemia, chronic lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, and multiple myeloma; the fibrotic diseases are pulmonary fibrosis, liver fibrosis, kidney fibrosis, or myocardial fibrosis.

12. A ligand, characterized in that, The ligand is a compound of formula A, or its enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts; Among them, R1 and R A L1 Same as the definition in claim 1.

13. The ligand as claimed in claim 12, characterized in that, The ligands are selected from compounds 1 to 16, 88 and 116-119 in Table A.

14. A degradation agent, characterized in that, The degrading agent is a conjugate formed by linking the ligand as described in claim 12 with a portion of the E3 ubiquitination enzyme ligand.

15. The degrading agent as described in claim 14, characterized in that, The degrading agent is a compound of formula B, or its enantiomers, diastereomers, racemates, or pharmaceutically acceptable salts; in, R2 is the ligand portion of the E3 ubiquitin ligase; L1 is a divalent linker; R B L1 Selected from the following group: none, optionally substituted C3-C10 carbocyclic, optionally substituted 3 to 10-membered heterocyclic, optionally substituted C6-C10 arylene, optionally substituted 5 to 10-membered heterocyclic, optionally substituted -C3-C10 carbocyclic-N(R3)-, optionally substituted -3 to 10-membered heterocyclic-N(R3)-, optionally substituted -C6-C10 arylene-N(R3)-, optionally substituted -5 to 10-membered heterocyclic-N(R3)-, optionally substituted -C3-C10 carbocyclic-O-, optionally substituted -3 to 10-membered heterocyclic-O-, optionally substituted -C6-C10 arylene-O- and optionally substituted -5 to 10-membered heterocyclic-O-; R3 is hydrogen or a C1-C4 alkyl group; "Optional substitution" and R1 are as defined in claim 1.

16. The degrading agent as described in claim 14, characterized in that, The degrading agents are selected from compounds 17 to 87 and compounds 89 to 115 in Table A.

17. A pharmaceutical composition comprising (i) the degrading agent as described in claim 14; and (ii) a pharmaceutically acceptable carrier.

18. Use of the degrading agent as claimed in any one of claims 14 to 16 or the pharmaceutical composition as claimed in claim 17 in the preparation of (i) a SMARCA2 protein inhibitor and / or (ii) a medicament for the prevention or treatment of diseases related to the activity or expression levels of SMARCA2 and SMARCA4 proteins.

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