WRN-targeted small molecule inhibitor and application thereof

Abstract

The invention relates to a small molecule inhibitor targeting WRN and application thereof. The invention discloses a compound shown in a formula I, or pharmaceutically acceptable salt or stereoisomer, isotope derivative or prodrug thereof, a pharmaceutical composition containing the compound, and a corresponding preparation method and application. Biological experiments show that the compound disclosed by the invention has a very strong WRN inhibition effect and can be used as a medicine for treating MSI tumors or MSI cancers.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of pharmaceutical chemistry, and particularly relates to a small-molecule inhibitor compound targeting WRN and application thereof. BACKGROUND

[0002] Cancer is originated from uncontrolled cell proliferation, which is related to genetic, age, environment and other factors, and essentially can be attributed to the result of gene mutation. These mutations are produced in normal life activities of cells such as genome replication, transcription, or DNA double-strand and single-strand breaks caused by other biological, physical and chemical means. In view of different types of DNA damage and spatiotemporal characteristics of cell life activities, cells have evolved complex and conservative DNA damage repair (DDR) mechanisms to maintain the stability of genetic material. The DNA mismatch repair (MMR) mechanism is to recognize and repair DNA damage caused by possible base insertion, deletion and mismatch during replication or recombination, which usually depends on several key protein factors MSH2, MSH6, MLH1 and PMS2. Mutation of these proteins will cause failure of DNA mismatch repair mechanism (MMRd), cause insertion or deletion of some simple repeat sequences-microsatellite on the genome, produce microsatellite instability (MSI) phenomenon, and cause the occurrence of Lynch Syndrome (LS) and other cancers. The clinical diagnosis scheme of MSI includes gene detection of MMR, immunohistochemistry (IHC) detection, or analysis of sequence changes of microsatellite sites BAT25, BAT26, D2S123, D5S346 and D17S250 by PCR, and according to different standards, tumors are divided into microsatellite stability (MSS), low-frequently microsatellite instability (MSI-L) and high-frequently microsatellite instability-high (MSI-H). MSI phenomenon accounts for about 3.8% of all tumor cases, about 15% of 1 / 2 grade solid tumors are MSI-H, and about 9% of 3 / 4 grade solid tumors are MSI-H, among which these cases contain about 30% of endometrial cancer, 20% of colorectal cancer and 20% of gastric cancer.

[0003] DNA double-strand break (DSB) repair is mainly achieved by homologous recombination (HR) and nonhomologus end joining (NHEJ). HR requires the participation of homologous DNA template, is more conservative and accurate, and mainly acts in S and G2 phases. NEHJ includes classic NHEJ (c-NHEJ) and alternative NHEJ (aNHEJ), and is involved in 75% of DSB repair and acts in all cell cycles. NHEJ does not require a template and can cause gene mutation and rearrangement, and is also the molecular basis of the great diversity of antibodies produced by VDJ in immune cells. In MSI cells, microsatellite repeat dinucleotide (TA) will accumulate and amplify, forming a cruciform secondary structure, interfering with DNA replication, and causing replication stress. Continuous replication stress can cause replication fork collapse, resulting in DNA double-strand breaks, causing cell apoptosis or becoming tumors, so the repair and restart of damaged replication forks and the repair of double-strand breaks are of great significance to maintain the stability of genetic material.

[0004] Werner Syndrome Helicase (WRN) is a member of the DNA helicase family, with a full length of 1432 amino acids. The N-terminal of the protein has 3'-5' DNA exonuclease activity, and the C-terminal has ATP-dependent 3'-5' DNA helicase activity. WRN can interact with proteins such as RPA, Ku, TRF2 and MRN complex, unwind DNA higher-order structures such as D-loops, holiday junction and G4, and is involved in DNA replication, telomere protection and double-strand break repair. The helicase activity of WRN is indispensable for the growth of MSI tumor cells. Knockout of WRN gene and inactivation mutation of the helicase activity center can inhibit the growth of MSI cell lines, and thus WRN is a synthetic lethality (SL) target for MSI tumors. Knocking out WRN in MSI tumor samples derived from patients resistant to chemotherapy or immune checkpoint inhibitors can also effectively inhibit the production of these tumor cells, indicating that WRN inhibitors have the potential for single-agent or combination therapy. Therefore, the present application provides a small molecule inhibitor targeting WRN as a safe and effective tumor-targeted therapeutic drug for microsatellite instability (MSI) cancer. SUMMARY

[0005] The present application relates to a small molecule inhibitor targeting WRN or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative or prodrug thereof, and its use in the treatment of tumors.

[0006] The present application relates to a small molecule inhibitor targeting WRN or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative or prodrug thereof, and its use in the treatment of tumors.

[0007] In a first aspect of the present application, there is provided a compound of Formula I or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative or prodrug thereof,

[0008]

[0009] In Formula I,

[0010] A represents a 5-6 membered aryl, saturated or partially unsaturated 5-6 membered heterocyclyl, 5-6 membered heteroaryl, 5-6 membered aryl and saturated or partially unsaturated 5-6 membered heterocyclyl, 5-6 membered aryl and 5-6 membered heteroaryl, saturated or partially unsaturated 5-6 membered heterocyclo and saturated or partially unsaturated 5-6 membered heterocyclyl, saturated or partially unsaturated 5-6 membered heterocyclo and 5-6 membered heteroaryl, 5-6 membered heteroaryl ring and saturated or partially unsaturated 5-6 membered heterocyclyl, or 5-6 membered heteroaryl ring and 5-6 membered heteroaryl, wherein ring A is optionally substituted with one or more groups;

[0011] L is an optionally substituted linking group selected from saturated or partially unsaturated straight chain C 1-13 heteroalkylene;

[0012] R1is selected from -C(O)-, -C(O)-CH2-, pyridinylidene, -NH-, -CH2-, and

[0013] R2, R3, R4, R5, and R6are each independently selected from the following groups:

[0014] -H, -OH, halogen (e.g., -F, -Cl, -Br, -I), -NO2, -COOH, and -B(OH)2;

[0015] optionally substituted C 1-5 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 cycloalkenyl, optionally substituted C 3-5 alkenyl, C 1-5 alkoxy, and -O-C 1-5 alkanoyl;

[0016] Optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups, wherein the optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups comprise one or more heteroatoms selected from O, N, or S, and when multiple heteroatoms are included, the heteroatoms may be identical or different; or

[0017] -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH, and -C. 1-5 Alkyl group;

[0018] Optionally, R4 is connected to one of R3 or R5 to form a saturated or unsaturated linear C that is optionally substituted. 1-5 Heteroalkyl group, wherein the heteroatom is selected from one or both of O or N;

[0019] Optionally, R1 and an adjacent group and their respective attached carbon atoms form a saturated or partially unsaturated 5-6 membered heterocycle containing one or two heteroatoms selected from O, N and S.

[0020] In some implementations, R1 is selected from...

[0021] In some implementations, L is derived from the formula "-D x -(CH2) p -(C(O)) y -(NR L ) z -(CH2) w -” indicates a linking group, where D is an atom selected from O and S, and x, y, and z are each independently selected from 0 or 1, w and p are each independently selected from integers from 0 to 10 and w + p ≤ 10, where R L Selected from -H, or optionally substituted C 1-5 Alkyl group. In some embodiments, w and p are each independently selected from integers from 0 to 5, for example, 0, 1, 2, 3, 4, or 5. In some embodiments, w is an integer selected from 0 to 5, for example, 0 to 3. In some embodiments, w is selected from 0 or 1.

[0022] In some embodiments, R2 and R6 are each independently selected from: -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), or -NH2, provided that R2 and R6 are both -H or R2 and R6 are different. In some embodiments, R2 and R6 are each independently selected from -F or -Br.

[0023] In some embodiments, R3 and R5 are each independently selected from the following groups, provided that R3 and R5 are both -H or -OH, or that R3 and R5 are different:

[0024] -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), -NO2, and -COOH;

[0025] Optional substitution of C 1-5 Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 Cycloalkenyl, optionally substituted C 3-5 alkenyl, C 1-5 Alkyl groups and -OC 1-5 Alkyl group;

[0026] Optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups, wherein the heterocyclic or heteroaryl group comprises one or more heteroatoms selected from O, N, or S, and when multiple heteroatoms are included, the heteroatoms may be identical or different; or

[0027] -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH, and -C. 1-5 Alkyl group.

[0028] In some embodiments, R3 and R5 are each independently selected from: optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted cycloheptyl, optionally substituted cyclooctyl, optionally substituted cyclononyl, optionally substituted cyclodecyl, optionally substituted cyclobutenyl, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cycloheptenyl, optionally substituted cyclooctenyl, optionally substituted cyclononenyl, optionally substituted cyclodecenyl, optionally substituted propenyl, optionally substituted butenyl, optionally substituted pentenyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-pentanoyl. In some embodiments, R3 and R5 are each independently selected from: -CH3 (-Me), -OCH3 (-OMe), -OC(O)CH3 (-OAc), -OC(O)CH2CH3, -C3H5 (e.g., isopropenyl), cyclopropyl, or cyclohexenyl substituted with -OH (e.g., ).

[0029] In some embodiments, R3 and R5 are each independently selected from: optionally halogen-substituted 5-6 membered heterocyclic groups or heteroaryl groups, wherein the heterocyclic group or heteroaryl group comprises one or more heteroatoms selected from O, N, or S. In some embodiments, R3 and R5 are each independently selected from: optionally halogen-substituted thiophene group, optionally halogen-substituted thiazolyl group, optionally halogen-substituted pyridinyl group, optionally halogen-substituted morpholinyl group, optionally halogen-substituted piperazine group. In some embodiments, R3 and R5 are each independently selected from:

[0030] In some implementations, R3 and R5 are each independently selected from: -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH and formyl, acetyl, propionyl, -butyryl or valeryl. In some embodiments, R3 and R5 are each independently selected from -NH2, -CONH2, or -CONHOH.

[0031] In some embodiments, R4 is selected from: -H, -OH, -B(OH)2, halogens (e.g., -F, -Cl, -Br, -I); optionally substituted C 1-5 Alkyl, C 1-5 Alkoxy, -OC 1-5 Alkyl group; or -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH, and -C. 1-5 Alkyl group.

[0032] In some embodiments, R4 is selected from: optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-pentanoyl. In some embodiments, R4 is selected from: -CH3 (-Me), -OCH3 (-OMe), -OCOCH3 (-OAc), or -OCOCH2CH3.

[0033] In some implementations, R4 is selected from: -NR a R b -C(O)NR a R b , where R a and Rb Each group is independently selected from -H, -OH, and formyl, acetyl, propionyl, -butyryl, or valeryl groups. In some embodiments, R4 is selected from -NH2 or -NHCOCH3 (-NHAc).

[0034] In some embodiments, R4 and R3 are connected to form a optionally substituted saturated or unsaturated linear C chain. 1-5 Heterohydrocarbon groups, in which the straight-chain C 1-5 The heteroalkylene group comprises one or two heteroatoms selected from O or N. In some embodiments, R4 and R3 are linked to form a heteroalkylene group selected from: -Q-optionally substituted C 1-5 Alkyl-Q- or -Q- optionally substituted C 1-5 The alkyl subunits are -Q, -QC(O)-Q-, where Q is independently O or N each time it appears. In some embodiments, the optionally substituted C... 1-5 The alkylene group is selected from optionally substituted methylene, ethylene, propylene, butylene, or pentylene. In some embodiments, the optionally substituted C 1-5 The alkyl subunit is selected from methyl, ethyl, propyl, butyl, or pentyl subunits. In some embodiments, the C-subunit is optionally substituted. 1-5 Alkylene or optionally substituted C 1-5 The alkyl subunit is replaced by -NH2. In some embodiments, the group formed by the connection of R4 and R3 is selected from...

[0035] In some embodiments, ring A is optionally replaced by R7 and / or R8, wherein,

[0036] R7 is selected from the following groups:

[0037] -H, =O, halogens (e.g., -F, -Cl, -Br, -I);

[0038] Optional substitution of C 1-5 Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 1-5 alkenyl, optionally substituted C 1-5 alkynyl group, optionally substituted C 1-5 Alkoxy, optional substituted C 1-5 Alkylene-C 1-5 Alkoxy;

[0039] Optionally substituted phenyl groups, or optionally substituted 5-6 membered heterocyclic groups or heteroaryl groups containing one or more heteroatoms selected from O, N, and S;

[0040] Optional substitution of C 1-5 Alkylene-NR c R d -NRc R d Where R c and R d Each is independently selected from -H or C. 1-5 Alkyl groups; and

[0041] C 1-5 alkylene-5-6-membered heterocyclic groups or C 1-5 Alkylene-5-6-membered heteroaryl, wherein the 5-6-membered heterocyclic group and the 5-6-membered heteroaryl group each independently comprise one or more heteroatoms selected from O, N, and S; and

[0042] R8 is selected from the following groups:

[0043] -H, halogens (-F, -Cl, -Br, -I);

[0044] Optional substitution of C 1-5 Alkyl, -C 1-5 Alkyl group; -C(O)-NR c R d , where R c and R d Each is independently selected from -H or C. 1-5 alkyl;

[0045] Optionally substituted phenyl groups, or optionally substituted 5-6 membered heterocyclic groups or heteroaryl groups containing one or more heteroatoms selected from O, N, and S;

[0046] Optionally substituted benzene with a 5-6 membered heterocyclic or heteroaryl group comprising one or more heteroatoms selected from O, N, and S; or optionally substituted 5-6 membered heterocyclic or heteroaryl cyclophenyl group comprising one or more heteroatoms selected from O, N, and S; or

[0047] Where Z represents O or S.

[0048] In some implementations, R7 is selected from one or more (e.g., 1, 2, 3, 4, or 5) R7s. 7s Replacement C 1-5 Alkyl group, optionally enclosed by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 3-10 Cycloalkyl, optionally enclosed by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 1-5 Alkenyl group, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 1-5 Alkyne group, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 1-5Alkyl group, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement -C 1-5 Alkylene-C 1-5 Alkyl group, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Substituted phenyl, wherein R 7s Selected from halogens, methyl, ethyl, propyl, butyl, pentyl, -CN, and phenyl.

[0049] In some implementations, R7 is selected from one or more (e.g., 1, 2, 3, 4, or 5) R7s. 7s Replacement C 1-5 Alkylene-NR c R d -NR c R d , where R c and R d Each is independently selected from -H, methyl, ethyl, propyl, butyl, and pentyl, wherein R 7s Selected from halogens, methyl, ethyl, propyl, butyl, pentyl, -CN, and phenyl.

[0050] In some implementations, R7 is selected from C. 1-5 Alkylene-pyridyl group.

[0051] In some implementations, R7 is selected from one or more (e.g., 1, 2, 3, 4, or 5) R7s. 7s Replacement One or two of Y1-Y3 are heteroatoms selected from N and O, and n is 1 or 2. This indicates that adjacent atoms on the ring form single bonds, double bonds, or the ring exhibits aromaticity, where R 7s The radical is selected from halogen, methyl, ethyl, propyl, butyl, pentyl, or cyano. In some embodiments, R7 is selected from those optionally mixed with one or more (e.g., 1, 2, 3, 4, or 5) R... 7s Replacement Where R 7s It is selected from halogen, methyl, ethyl, propyl, butyl, pentyl or cyano.

[0052] In some embodiments, R7 is optionally selected from: -F, =O, methyl, ethyl, propyl (e.g., n-propyl, isopropyl), cyclopropyl, cyclopentyl, cyclohexyl, phenyl, benzyl, dimethylaminoethyl, methoxyethyl, methylaminoethyl, cyanomethyl, allyl, cyanophenyl.

[0053] In some implementations, R7 is optionally selected from:

[0054] In some embodiments, R8 is selected from the following groups:

[0055] Optional by one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 or R 8S2 Substituted phenyl groups; optionally substituted with one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 or R 8S2 The substituted 5-6 membered heterocyclic or heteroaryl group, optionally with one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Substituted benzo5-6-membered heterocyclic or heteroaryl groups, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 and / or R 8S2 Substituted 5-6 membered heterocyclic or heteroaromatic phenyl groups, optionally with one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 and / or R 8S2 Substituted 5-6 membered heterocyclic or heteroaryl rings and 5-6 membered heterocyclic or heteroaryl groups, wherein R 8S1 Selected from: -H, -OH, =O, halogens (-F, -Cl, -Br, -I), -CN, -COOH, C 1-10 Alkyl, C 1-5 Alkoxy, C 1-5 Haloalkyl, C 1-5 Alkyl group, C 1-5 Alkyl group, -C(O)NR e R f C(O)NR e R f -(CH2) q -NR e R f -SO2R g -NR e R f -C 1-5 Alkyl group, wherein R e and R f Selected from -H, C 1-5 Alkyl groups, and R g Selected from C 1-5 Alkyl and amino groups, and R 8S2 Selected from 5-6 member saturated or partially unsaturated heterocyclic ketone groups, 5-6 member saturated or partially unsaturated heterocyclic groups.

[0056] In some implementations, R 8S2 It is selected from 5-6 member saturated or partially unsaturated heterocyclic ketone groups, 5-6 member saturated or partially unsaturated heterocyclic groups, wherein the heterocycle contains one or more heteroatoms selected from O, N and S.

[0057] In some implementations, R 8S2 Optionally R 8S2a Replace, where R 8S2a Selected from C 1-5 alkyl.

[0058] In some implementations, R 8S2 Selected from: In some implementations, R 8S2 Selected from:

[0059] In some implementations, R8 is selected from...

[0060] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 The substituted 5-6 membered heterocyclic or heteroaryl group, optionally with one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Substituted benzo5-6-membered heterocyclic or heteroaryl groups, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 or R 8S2 A substituted 5-6 membered heterocyclic or heteroaryl phenyl group, wherein the 5-6 membered heterocyclic or heteroaryl group or the 5-6 membered heterocyclic or heteroaryl ring contains one or more heteroatoms selected from O, N and S.

[0061] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 Replaced by: In U1-U5, one or two of the elements are independently selected from one or two of O, N, or S, and the rest are carbon; in V1-V6, one or two of the elements are independently selected from N or O, and the rest are carbon. This indicates that adjacent atoms on the ring are connected by single bonds, double bonds, or that the ring is aromatic.

[0062] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement

[0063] In some implementations, R8 is selected from:

[0064] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement Where Q is CH or N, T1 and T2 are independently one of N, S, O or C, T3 and T4 are independently one of N, S, O or C, and n is 1 or 2. This indicates that single or double bonds are formed between adjacent atoms on the ring, or that the ring is aromatic.

[0065] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement

[0066] In some implementations, R8 is selected from:

[0067] In some implementations, ring A represents rings that are optionally replaced by R7 and / or R8: R7 and R8 are as described above. Among them, 1-3 of G1-G5 are independently selected from N or O, and the rest are C; 1 to 2 of I1-I4 are N, and the rest are C; 1 to 3 of J1-J6 are N, and the rest are C; 1 to 2 of K1-K3 are N, and the rest are C, and 0 to 2 of K4-K7 are N, and the rest are C; 1 to 2 of L1-L3 are N, and the rest are C; 1 of M1-M2 is S, and the rest are C. This indicates that adjacent atoms on the ring are connected by single bonds, double bonds, or that the ring is aromatic.

[0068] In some implementations, ring A represents: In this configuration, one to three of G1, G3, and G4 are N, and the rest are CH. In some implementations, ring A represents: In this system, one of G1, G3, G4, and G5 is 0, one is N, and the rest are C. In some implementations, ring A represents: One of G1, G3, G4, and G5 is N, and the rest are C.

[0069] In some implementations, ring A represents And R7 and R8 are as described above.

[0070] In some implementations, ring A represents And R7 and R8 are as described above.

[0071] In some implementations, ring A represents Of J2-J6, 1 to 3 are N, and the rest are C. In some implementations, ring A represents: Of these, one or two of J2-J6 are N, and the rest are C. In some implementations, ring A represents: And R7 and R8 are as described above.

[0072] In some implementations, ring A represents In this configuration, one of K1-K2 is N, and the rest are C; and in K4-K7, 0 to 2 are N, and the rest are C. In some implementations, ring A represents: In this configuration, one or two of K1 and K3 are N, and the rest are C; and in K4-K7, one or two are N, and the rest are C. In some implementations, ring A represents: R7 and R8 are as described above.

[0073] In some implementations, ring A represents

[0074] In some embodiments, a compound represented by Formula II, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, is provided.

[0075]

[0076] Wherein rings A, L, R2, R3, R4, and R5 are defined as described above, and ring B represents a 5-6 membered heterocycle containing one or two saturated or partially unsaturated heterocycles selected from O, N, and S heteroatoms.

[0077] In some embodiments, the compounds of formula II have the structures shown in formulas II-1 to II-4 as follows:

[0078]

[0079] Where Q appears independently as either N or CH each time. This indicates that single or double bonds are formed between adjacent atoms on the ring, or that the ring is aromatic, and that R9 is selected from: -H, halogens (e.g., -F, -Cl, -Br, -I), C 1-5 Haloalkyl, C 1-5 Alkyl alcohol, optionally substituted phenyl, or C 1-5 Alkylene-C(O)NR 9a R 9b , where R 9aand R 9b Selected from -H, C 1-5 Alkyl group. In some embodiments, R9 is a halogen (e.g., -F, -Cl, -Br, -I), -CH3, -C3H7 (e.g., - i Pr), -C4H9 (e.g., - n Bu), -CH2CHF2, -CH2CH2OH, -CH2C(O)NMe2, -CH2C(O)NH2. In some embodiments, R9 is selected from optionally substituted phenyl groups, wherein the optionally substituted phenyl group is substituted by one or more C2O4 groups. 1-5 Halogenated alkyl groups or halogens (e.g., -F, -Cl, -Br, -I) are substituted. In some embodiments, C 1-5 The haloalkyl group is -CF3. In some embodiments, the optionally substituted phenyl group is...

[0080] In some embodiments, the compounds represented by Formula II-1 or Formula II-2 have structures represented by the following formulas:

[0081]

[0082] In some embodiments, the compound represented by Formula II has a structure represented by the following formula:

[0083]

[0084] In some embodiments, R2 is selected from -H or halogens (e.g., -F, -Cl, -Br, -I). In some embodiments, R2 is selected from -Br.

[0085] In some embodiments, R3 and R4 are each independently selected from: -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), -OC 1-5 Alkyl group. In some embodiments, R3 and R4 are linked to form a heteroalkylene group, wherein the heteroatom is selected from O or N. In some embodiments, the group formed by the linkage of R4 and R3 is selected from...

[0086] In some embodiments, R5 is selected from: -H, -OH, halogens (e.g., -F, -Cl, -Br, -I); optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 Cycloalkenyl, optionally substituted C 3-5 Alkenyl; optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups, wherein the heterocyclic or heteroaryl group contains one or more heteroatoms selected from O, N or S, and when multiple heteroatoms are contained, the heteroatoms may be the same or different.

[0087] In some embodiments, R5 is selected from: optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted cycloheptyl, optionally substituted cyclooctyl, optionally substituted cyclononyl, optionally substituted cyclodecyl, optionally substituted cyclobutenyl, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cycloheptenyl, optionally substituted cyclooctenyl, optionally substituted cyclononenyl, optionally substituted cyclodecenyl, optionally substituted propenyl, optionally substituted butenyl, optionally substituted pentenyl. In some embodiments, R5 is each independently selected from: -C3H5 (e.g., isopropenyl), cyclopropyl, or cyclohexenyl substituted with -OH (e.g., ...). ).

[0088] In some embodiments, R5 is selected from: optionally halogen-substituted 5-6 membered heterocyclic groups or heteroaryl groups. In some embodiments, R5 is selected from: optionally halogen-substituted thiophene group, optionally halogen-substituted thiazolyl group, optionally halogen-substituted pyridinyl group, optionally halogen-substituted morpholinyl group, optionally halogen-substituted piperazine group. In some embodiments, R5 is independently selected from:

[0089] In some embodiments, the compound represented by Formula I has the following structure: Rings A, L, R2, R3, R4, R5, and R6 are defined as above.

[0090] In some embodiments, R2 and R6 are each independently selected from: -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), provided that R2 and R6 are both -H or R2 and R6 are different. In some embodiments, R2 and R6 are each independently selected from -F or -Br.

[0091] In some embodiments, R3 and R5 are each independently selected from the following groups, provided that R3 and R5 are both -H or -OH, or that R3 and R5 are different:

[0092] -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), -NO2, and -COOH;

[0093] Optional substitution of C 1-5 Alkyl, C 1-5 Alkyl groups and -OC 1-5 Alkyl group;

[0094] -NR a R b -C(O)NR a R b , where R a and Rb Each is independently selected from -H and -OH.

[0095] In some embodiments, R3 and R5 are each independently selected from: optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-pentanoyl. In some embodiments, R3 and R5 are each independently selected from: -CH3 (-Me), -OCH3 (-OMe), -OC(O)CH3 (-OAc).

[0096] In some implementations, R3 and R5 are each independently selected from: -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H and -OH. In some embodiments, R3 and R5 are independently selected from -NH2, -CONH2, or -CONHOH.

[0097] In some embodiments, R4 is selected from: -H, -OH, -B(OH)2, halogens (e.g., -F, -Cl, -Br, -I); optionally substituted C 1-5 Alkyl, C 1-5 Alkoxy, -OC 1-5 Alkyl group; or -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH, and -C. 1-5 Alkyl group.

[0098] In some embodiments, R4 is selected from: optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-pentanoyl. In some embodiments, R4 is selected from: -CH3 (-Me), -OCH3 (-OMe), -OCOCH3 (-OAc), or -OCOCH2CH3.

[0099] In some implementations, R4 is selected from: -NR a R b -C(O)NR a R b, where R a and R b Each group is independently selected from -H, -OH, and formyl, acetyl, propionyl, -butyryl, or valeryl groups. In some embodiments, R4 is selected from -NH2 or -NHCOCH3 (-NHAc).

[0100] In some embodiments, R4 is connected with R3 or R5 to form a optionally substituted saturated or unsaturated linear C chain. 1-5 Heterohydrocarbon groups, in which the straight-chain C 1-5 The heteroalkylene group contains one or both heteroatoms selected from O or N. In some embodiments, R4 and R3 are linked to form a group selected from: -Q-optionally substituted C 1-5 Alkyl-Q- or -Q- optionally substituted C 1-5 The alkyl subunit is -Q, -C(O)-, where Q is independently O or N each time it appears. In some embodiments, the optionally substituted C... 1-5 The alkylene group is selected from optionally substituted methylene, ethylene, propylene, butylene, or pentylene. In some embodiments, the optionally substituted C 1-5 The alkyl subunit is selected from methyl, ethyl, propyl, butyl, or pentyl subunits. In some embodiments, the group formed by the connection of R4 and R3 is selected from...

[0101] In some embodiments, a compound represented by formula IV-1 or IV-2, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, is provided.

[0102] R2, R3, R4, R5, R7, R8 and R9 have the same definitions as those defined with respect to Equation I or Equation II, and X is N or CN.

[0103] In some embodiments, the compounds represented by formula IV-1 or IV-2 have the following structures:

[0104]

[0105] In some embodiments, R2 is selected from -H or halogens (e.g., -F, -Cl, -Br, -I). In some embodiments, R2 is selected from -Br.

[0106] In some embodiments, R3 and R4 are each independently selected from: -H, -OH, halogens (e.g., -F, -Cl, -Br, -I), -OC 1-5 Alkyl group. In some embodiments, R3 and R4 are linked to form a heteroalkylene group, wherein the heteroatom is selected from O or N. In some embodiments, the group formed by the linkage of R4 and R3 is selected from...

[0107] In some implementations, R5 is selected from: -H, halogen (e.g., -F, -Cl, -Br, -I).

[0108] In some implementations, R9 is selected from: -H, C 1-5 Alkyl group. In some embodiments, R9 is -CH3.

[0109] In some embodiments, R7 is selected from the following groups:

[0110] -H;

[0111] Optional substitution of C 1-5 Alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 1-5 alkenyl;

[0112] Optionally substituted phenyl groups, optionally substituted 5-6 membered heterocyclic groups, or heteroaryl groups; and

[0113] C 1-5 Alkylene-5-6-membered heterocyclic or heteroaryl groups.

[0114] In some implementations, R7 is selected from one or more (e.g., 1, 2, 3, 4, or 5) R7s. 7s Replacement C 1-5 Alkyl group, optionally enclosed by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 3-10 Cycloalkyl, optionally enclosed by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Replacement C 1-5 Alkenyl group, optionally surrounded by one or more (e.g., 1, 2, 3, 4, or 5) R 7s Substituted phenyl, wherein R 7s Selected from halogens, methyl, ethyl, propyl, butyl, pentyl, -CN, and phenyl.

[0115] In some implementations, R7 is selected from C. 1-5 Alkylene-pyridyl group.

[0116] In some implementations, R7 is selected from one or more (e.g., 1, 2, 3, 4, or 5) R7s. 7s Replacement In Y1-Y3, one or two heteroatoms are independently selected from N and O, and n is 1 or 2. This indicates that adjacent atoms on the ring form single bonds, double bonds, or the ring exhibits aromaticity, where R 7sThe radical is selected from halogen, methyl, ethyl, propyl, butyl, pentyl, or cyano. In some embodiments, R7 is selected from those optionally mixed with one or more (e.g., 1, 2, 3, 4, or 5) R... 7s Replacement Where R 7s It is selected from halogen, methyl, ethyl, propyl, butyl, pentyl or cyano.

[0117] In some embodiments, R7 is optionally selected from: methyl, ethyl, propyl (e.g., n-propyl, isopropyl), cyclopropyl, cyclopentyl, cyclohexyl, phenyl, benzyl, cyanomethyl, allyl, cyanophenyl.

[0118] In some implementations, R7 is optionally selected from:

[0119] R8 is selected from the following groups:

[0120] Optionally substituted phenyl groups, optionally substituted 5-6 membered heterocyclic groups, or heteroaryl groups; or

[0121] Optional substituted benzo5-6 membered heterocyclic groups or heteroaryl groups.

[0122] In some embodiments, R8 is selected from the following groups:

[0123] Optional by one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 or R 8S2 Substituted phenyl groups; optionally substituted with one or more (e.g., 1, 2, 3, 4, or 5) R 8S1 or R 8S2 The substituted 5-6 membered heterocyclic or heteroaryl group, optionally with one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Substituted benzo5-6-membered heterocyclic or heteroaryl groups, wherein R 8S1 Selected from: -H, -OH, halogens (-F, -Cl, -Br, -I), -COOH, C 1-10 Alkyl, C 1-5 Haloalkyl, C 1-5 alkyl ester group, and R 8S2 Selected from 5-6 member saturated or partially unsaturated heterocyclic groups, wherein the heterocycle contains one or more heteroatoms selected from O, N and S.

[0124] In some implementations, R 8S2 for

[0125] In some implementations, R8 is selected from:

[0126] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 The substituted 5-6 membered heterocyclic or heteroaryl group, optionally with one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 The substituted benzo5-6-membered heterocyclic group or heteroaryl group, wherein the 5-6-membered heterocyclic group or heteroaryl group or 5-6-membered heterocyclic group or heteroaryl group contains one or more heteroatoms selected from O, N and S.

[0127] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 Replaced by: In U1-U5, one or two of the elements are independently selected from one or two of N, O and S, and the rest are carbon; in V1-V6, one or two of the elements are independently selected from N or O, and the rest are carbon. This indicates that adjacent atoms on the ring are connected by single bonds, double bonds, or that the ring is aromatic.

[0128] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement

[0129] In some implementations, R8 is selected from:

[0130] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement Where Q is CH or N, T1 and T2 are independently one of N, S, O or C, T3 and T4 are independently one of N, S, O or C, and n is 1 or 2. This indicates that single or double bonds are formed between adjacent atoms on the ring, or that the ring is aromatic.

[0131] In some implementations, R8 is selected from: optionally selected by one or more (e.g., 1, 2, 3, 4 or 5) R 8S1 or R 8S2 Replacement

[0132] In some implementations, R8 is selected from:

[0133] In some embodiments, a compound represented by formula V-1 or V-2, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, is provided.

[0134] Among them, R2, R3, R4, R5, R7, R8 and R9 have the same definition as defined with respect to Equation I or Equation II, one of K1 and K2 is N and the other is CR7; one of K4 to K7 is CR8, 0-2 are N, and the rest are CH; This indicates that adjacent atoms on the ring are connected by single bonds, double bonds, or that the ring is aromatic.

[0135] In some embodiments, the compound represented by formula V-1 or formula V-2 is the compound shown in the formula:

[0136]

[0137] In some embodiments, R2 is selected from -H or halogens (e.g., -F, -Cl, -Br, -I). In some embodiments, R2 is selected from -Br.

[0138] In some implementations, R3 and R4 are each independently selected from -H and -OH.

[0139] In some embodiments, R5 is selected from: -H, halogens (e.g., -F, -Cl, -Br, -I); optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 Cycloalkenyl, optionally substituted C 3-5 Alkenyl; optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups, wherein the heterocyclic or heteroaryl group contains one or more heteroatoms selected from O, N or S, and when multiple heteroatoms are contained, the heteroatoms may be the same or different.

[0140] In some embodiments, R5 is selected from: optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted cycloheptyl, optionally substituted cyclooctyl, optionally substituted cyclononyl, optionally substituted cyclodecyl, optionally substituted cyclobutenyl, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cycloheptenyl, optionally substituted cyclooctenyl, optionally substituted cyclononenyl, optionally substituted cyclodecenyl, optionally substituted propenyl, optionally substituted butenyl, optionally substituted pentenyl. In some embodiments, R5 is each independently selected from: -C3H5 (e.g., isopropenyl), cyclopropyl, or cyclohexenyl substituted with -OH (e.g., ...). ).

[0141] In some embodiments, R5 is selected from: optionally halogen-substituted 5-6 membered heterocyclic groups or heteroaryl groups. In some embodiments, R5 is selected from: optionally halogen-substituted thiophene group, optionally halogen-substituted thiazolyl group, optionally halogen-substituted pyridinyl group, optionally halogen-substituted morpholinyl group, optionally halogen-substituted piperazine group. In some embodiments, R5 is independently selected from:

[0142] In the following implementation, R7 is selected from: -H, C 1-5 Alkyl group. In some embodiments, R7 is selected from methyl groups.

[0143] R8 is selected from the following groups: -H, halogens (-F, -Cl, -Br, -I); -C(O)-NH2.

[0144] In some embodiments, the compounds of the present invention are selected from the group consisting of the compounds in Table 1, as well as their pharmaceutically acceptable salts, stereoisomers, isotope derivatives, or prodrugs.

[0145] Table 1: Exemplary Compounds of the Invention

[0146]

[0147]

[0148]

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155] A second aspect of the present invention is to provide a pharmaceutical composition comprising a compound provided according to a first aspect of the present invention, or a pharmaceutically acceptable salt, stereoisomer, isotope derivative or prodrug thereof, and a pharmaceutically acceptable carrier or excipient.

[0156] A third aspect of the invention is to provide the use of a compound provided according to the first aspect of the invention, or a pharmaceutically acceptable salt, stereoisomer, isotope derivative or prodrug thereof, in the preparation of a medicament or kit for reducing WRN activity in cells or in a subject in need.

[0157] In some embodiments, the cells are derived from proliferative diseases, preferably cancer cells, and more preferably, MSI cancer cells.

[0158] In some embodiments, the subject suffers from a proliferative disease, preferably cancer, and more preferably, MSI cancer.

[0159] In some implementations, the MSI cancer is either a low-frequently microsatellite instability (MSI-L) cancer or a high-frequently microsatellite instability (MSI-H) cancer.

[0160] In some implementations, the MSI cancer is characterized by high microsatellite instability.

[0161] In some embodiments, the MSI cancer is selected from colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and / or prostate cancer.

[0162] In some embodiments, the subject is a mammal, preferably a human or a veterinary animal.

[0163] A fourth aspect of the invention is to provide a method for reducing the activity of WRN in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a compound provided according to a first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, isotope derivative or prodrug thereof, or a pharmaceutical composition provided according to a second aspect of the invention.

[0164] In some embodiments, the subject suffers from a proliferative disease, preferably cancer, and more preferably, MSI cancer.

[0165] In some embodiments, the MSI cancer is a microsatellite low-instability MSI-L cancer or a microsatellite high-instability MSI-H cancer.

[0166] In some implementations, the MSI cancer is characterized by high microsatellite instability.

[0167] In some embodiments, the MSI cancer is selected from colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and / or prostate cancer.

[0168] In some embodiments, the subject is a mammal, preferably a human or a veterinary animal.

[0169] A fifth aspect of the invention is to provide a method for treating proliferative diseases in a subject in need, the method comprising administering to the subject a therapeutically effective amount of a compound provided according to a first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, isotope derivative or prodrug thereof, or a pharmaceutical composition provided according to a second aspect of the invention.

[0170] In some embodiments, the proliferative disease is cancer, preferably, the cancer is MSI cancer.

[0171] In some embodiments, the MSI cancer is a microsatellite low-instability MSI-L cancer or a microsatellite high-instability MSI-H cancer.

[0172] In some implementations, the MSI cancer is characterized by high microsatellite instability.

[0173] In some embodiments, the MSI cancer is selected from colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and / or prostate cancer.

[0174] In some embodiments, the subject is a mammal, preferably a human or a veterinary animal.

[0175] A sixth aspect of the invention is to provide a method for reducing the activity of WRN in MSI cancer cells, the method comprising contacting the cells with a compound provided according to a first aspect of the invention or a pharmaceutically acceptable salt, stereoisomer, isotope derivative or prodrug thereof, or a pharmaceutical composition provided according to a second aspect of the invention.

[0176] In some embodiments, the MSI cancer cells are characterized by low microsatellite instability (MSI-L) or high microsatellite instability (MSI-H).

[0177] In some embodiments, the MSI cancer cells are mammalian cells. In some embodiments, the method is an in vitro, non-therapeutic method. In some embodiments, the MSI cancer cells are in vivo in a subject.

[0178] In some embodiments, the animal is a mammal, preferably a human or veterinary animal. Detailed Implementation

[0179] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly used in the field to which this invention pertains. For the purposes of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural forms, and vice versa.

[0180] definition

[0181] As described herein, the compounds of this invention can be expanded by substituting any number of substituents or functional groups. Generally, when used in general formulas including substituents in this invention, the term "substitution," whether preceding or following the term "optional," refers to replacing a hydrogen radical with a substituent of a specified structure. When multiple positions in a particular structure are substituted by multiple specific substituents, each substituent position can be the same or different. The term "substitution" as used herein includes all permissible substitutions in organic compounds. In a broad sense, permissible substituents include acyclic, cyclic, branched, unbranched, carbocyclic, and heterocyclic, aromatic and non-aromatic organic compounds. In this invention, heteroatomic nitrogen may be supplemented with hydrogen substituents or any permissible substituents to complement its valence state. Furthermore, this invention considers the combination of substituents and variable groups to provide a stable form of compound, thereby facilitating the treatment of diseases. The term "stable" here means a compound having a stable structure that is sufficient to maintain the integrity of the compound structure when detected for a sufficiently long period of time, preferably remaining effective for a sufficiently long period of time.

[0182] The term "alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having a specified number of carbon atoms. Alkyl groups used in this invention (including those themselves or as a portion of another substituent) may contain 1 to 20 carbon atoms, for example 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms, preferably 1 to 15 carbon atoms, more preferably 1 to 10 carbon atoms, and even more preferably 1 to 5 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and their various branched isomers. Further, the term "C m-n "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having m or n carbon atoms. For example, "C 1-8 "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having 1 to 8 carbon atoms, for example, a straight-chain or branched saturated aliphatic hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms.

[0183] The term "alkenyl" refers to a straight-chain or branched, non-cyclic unsaturated hydrocarbon group having a specified number of carbon atoms, wherein at least two carbon atoms are bonded together by unsaturated double bonds. The alkenyl groups suitable for use in this invention can have 2-10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) carbon atoms, preferably 2-5 carbon atoms. Further, the term "C..." m-n "Alkenyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having mn carbon atoms. For example, "C 2-10 "Alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group having 2-10 carbon atoms, for example, a straight-chain or branched saturated aliphatic hydrocarbon group having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. 2-5 Examples of alkenyl groups include, but are not limited to, vinyl, 1-propen-1-yl, 1-propen-2-yl, 2-propen-1-yl, 2-methyl-1-propen-1-yl, 2-methyl-2-propen-1-yl, 1-buten-1-yl, 1-buten-2-yl, 2-buten-1-yl, 2-buten-2-yl, 3-buten-1-yl, 3-buten-2-yl, 1,3-butadien-1-yl, 1,3-butadien-2-yl, 1-penten-1-yl, 2-penten-1-yl, 2-penten-2-yl, 3-penten-1-yl, 3-penten-3-yl, 4-penten-1-yl, 4-penten-4-yl, etc. Preferably, the alkenyl group has a double bond. It is also preferred that the double bond in the alkenyl group is directly connected to the rest of the compound containing the alkenyl group. For example, 1-propen-1-yl is more preferred than 2-propen-1-yl, whereby when the substituent is connected to ring A or ring B, it is possible to form an extended conjugated system.

[0184] The term "cycloalkyl" refers to cyclic alkyl groups comprising saturated monocyclic, bicyclic, or polycyclic rings, such as 3-10 membered cycloalkyl groups, for example, monocyclic, bicyclic, or polycyclic rings having 3, 4, 5, 6, 7, 8, 9, or 10 carbon ring atoms. Cycloalkyl groups may also include spirocyclic, bridged, and fused ring structures, such as 5-10 membered fused bicyclic rings and 5-10 membered spirobicyclic rings. Cycloalkyl groups suitable for use in this invention include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and norbornel. Examples of 5-10 membered fused bicyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and norbornel. Examples of 5-10 inch double-ring screws include, but are not limited to, those shown below. In this invention, the cycloalkyl group is preferably a monocyclic cycloalkyl group containing 3 to 10 carbon ring atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0185] The term "cycloalkenyl" refers to a cycloalkyl group containing one or more unsaturated double bonds, such as a cycloalkenyl group containing 3 to 10 carbon ring atoms, for example, a monocyclic, bicyclic, or polycyclic cycloalkenyl group having 3, 4, 5, 6, 7, 8, 9, or 10 carbon ring atoms. The cycloalkenyl groups suitable for use in this invention may have 4 to 8 carbon ring atoms, for example, cyclohexene.

[0186] Unless otherwise explicitly specified, the term "heteroatom" as used herein refers to an oxygen atom, a nitrogen atom, or a sulfur atom. For example, "optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups comprising one or more heteroatoms" means "optionally substituted 5-6 member saturated or partially unsaturated heterocyclic or heteroaryl groups comprising one or more atoms selected from oxygen, nitrogen, or sulfur." ​​When explicitly specified, "heteroatom" refers only to the specifically designated atom; for example, in "one or two of V1-V6 selected from N or O," the "heteroatom" can only be nitrogen or oxygen.

[0187] The term "heteroalkylene group" itself, or as part of another substituent, refers to a divalent group containing a heteroatom, such as, but not limited to, -S-CH2-CH2-CH2-CH2-, -O-CH2-CH2-CH2-CH2-, -S-CH2-CH2-NR-CH2-CH2-, -O-CH2-CH2-O-, and -N-CH=N-. For heteroalkylene groups, the heteroatom can also occupy any one or both of the chain ends (e.g., alkyleneoxy, alkylenedioxy, alkyleneamino, alkylenediamino, etc.). Furthermore, for the heteroalkylene linking group, the direction of the formula for the linking group does not indicate the orientation of the linking group. For example, the formula -C(O)2R'- represents both -C(O)2R'- and -R'C(O)2-. In this document, the heteroalkylene group is fully saturated or partially unsaturated, and its main chain can contain one or more double or triple bonds.

[0188] The terms "heterocyclic," "heterocyclic group," or "heterocyclic alkyl" refer to substituted or unsubstituted non-aromatic, partially unsaturated, or fully saturated heterocyclic groups. Heterocyclic groups may contain 1 to 3 heteroatoms selected from N, O, or S as ring members, and may include monocyclic heterocycles, bicyclic bridged heterocycles, bicyclic fused heterocycles, and bicyclic spirocyclic heterocycles, etc. Heterocyclic alkyl groups suitable for this invention may contain 3 to 12 ring atoms (3-12 membered heterocycles), for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, preferably 3-12, and even more preferably 4-10 or 4-8 ring atoms. Heterocyclic alkyl groups may be attached to heteroatoms or carbon atoms. The N and S in the heterocyclic alkyl ring may be oxidized to various oxidation states. Non-limiting examples of heterocyclic alkyl groups include oxoheterobutyl, azaheterobutyl, tetrahydrofuranyl, tetrahydropyrroleyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiaranyl, dioxaneyl, piperazineyl, hexahydropyrazineyl, morpholinyl, piperidinyl, dithiaalkyl, etc. In general, the terms "heterocyclic," "heterocyclic group," or "heterocyclic alkyl" are used interchangeably. In certain specific cases, the terms "heterocyclic" and "heterocyclic group" are used to describe different parts of a group in which the group is connected to the main structure of the compound via the "heterocyclic group." For example, a 5-membered heterocyclic fused to a 6-membered heterocyclic group indicates that the group is connected to the main structure of the compound via atoms on the 6-membered heterocyclic group.

[0189] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic ring system (preferably 5-10 membered aromatic rings) having a total of 5 to 15 ring members (or ring atoms), wherein all rings in the system are aromatic and each ring in the system contains 3 to 7 ring members; when not all rings in the system are aromatic, it is considered a cycloalkyl or heterocyclic alkyl group. In some embodiments of the invention, "aryl" refers to an aromatic ring system, including but not limited to phenyl, biphenyl, indanyl, 1-naphthyl, 2-naphthyl, tetrahydronaphthyl, anthracene, and phenanthrene. Fused aryl groups may be attached to another group at a suitable position on a cycloalkyl or aromatic ring. The term "aromaticity" refers to the fact that the electrons of the atoms in the ring structure form a conjugated system and the number of electrons conforms to Hückel's rule.

[0190] The terms "heteroaromatic ring" or "heteroaryl" refer to a heterocycle or heterocyclic group that contains aromatic compounds. In this invention, 5- or 6-membered heterocycles or heteroaryl groups are preferred. Non-limiting examples of heteroaryl groups include thiophene, furanyl, pyrrole, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, thiazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl. In this application, the terms "heteroaromatic ring" and "heteroaryl" are used interchangeably. In certain specific cases, the terms "heteroaromatic ring" and "heteroaryl" are used to describe different parts of a group in which the group is linked to the main structure of the compound via the "heteroaryl" group. For example, a 5-membered heteroaromatic ring fused to a 6-membered heteroaryl indicates that the group is linked to the main structure of the compound via atoms on the 6-membered heteroaryl group.

[0191] The term "fused-ring heteroaryl" refers to a plurality of rings fused together, wherein at least one of the fused rings is a heteroaryl ring. In this document, "fused-ring heteroaryl" is also described as benzo[a]heteroaryl (heterocyclic) or heteroaryl[a]phenyl. When described as "benzo[a]heteroaryl (heterocyclic)", it is intended to indicate that the bond to which the group is attached to the remainder of the compound is located on a heteroaryl ring; when described as "heteroaryl[a]phenyl[a]phenyl", it is intended to indicate that the bond to which the group is attached to the remainder of the compound is located on a benzene ring. Fused-ring heteroaryl groups suitable for use in this invention include 5,6-fused-ring heteroaryl, 6,6-fused-ring heteroaryl, or 6,5-fused-ring heteroaryl. Non-limiting examples of fused-ring heteroaryl groups include fluorenyl, quinolinyl, isoquinolinyl, quinazolinyl, quinoxalolinyl, cyclophosphinyl, naphthidyl, triazineyl, indoleyl, indazoleyl, benzofuranyl, benzothiophenyl, benzooxazolyl, benzotriazolyl, and benzothiazolyl. Heteroaryl or fused-ring heteroaryl groups can be attached to the rest of the molecule via carbon atoms or heteroatoms.

[0192] The term "alkoxy" or "oxoalkyl" refers to -O-alkyl, such as -OC. 1-10 Alkyl group, preferably -OC 1-5 Alkyl, more preferably -OC 1-3 Alkyl groups. Non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy, and cyclobutoxy.

[0193] The term "alkanoyl" or "acylalkyl" refers to -C(O)-alkyl or -C(O)-H (formyl or aldehyde), for example -C(O)-C 1-10 Alkyl group, preferably --C(O)-C 1-5 Alkyl groups, more preferably -C(O)-C 1-3Alkyl groups, such as -C(O)-CH3 (acetyl). Non-limiting examples of alkyl groups include acetyl, n-propionyl, isopropionyl, n-butyryl, sec-butyryl, isobutyryl, tert-butyryl, n-valeryl, n-hexanoyl, cyclopropionyl, and cyclobutyryl.

[0194] The terms "alkylidene" or "alkylidene" or "alkylenetriole" refer to an alkyl group that is attached to the molecular backbone by two single bonds, for example, -alkyl-, such as -C. 1-10 Alkyl groups, where an alkyl group is attached to the same atom of the molecular backbone by two single bonds, are called "alkylidene" or "alkylene"; where an alkyl group is attached to two separate atoms of the molecular backbone by two separate single bonds, the alkyl group is called "alkylidene" or "alkylene". In this document, the terms "alkylidene", "alkylene", "alkylidene", or "alkylene" are used interchangeably. Non-limiting examples of alkylidene groups include methyleneidene (-CH2-), ethoxyidene (-C2H4-), propyleneidene (-C3H6-), butylidene (-C4H8-), and hexylidene (-C5H6-). 10 -).

[0195] The term "alkyl subunit" refers to an alkyl group that is attached to the molecular skeleton by one single bond and one double bond, for example, -alkyl=, such as -C 1-10 Alkyl group = . Non-limiting examples of alkyl subunits include methyl subunit (-CH=), ethyl subunit (-C2H3=), propyl subunit (-C3H5-), butyl subunit (-C4H7-), and hexyl subunit (-C5H9-).

[0196] The term "halogen" refers to F, Cl, Br, I, or their isotopes.

[0197] The term "halogenation" or "halogen substitution" refers to the substitution of a hydrogen atom by one or more isotopes selected from F, Cl, Br, I, or their isotopes. The upper limit of the number of halogen substituents is equal to the sum of the number of hydrogen atoms that can be substituted in the substituted group. Unless otherwise specified, the number of halogen substituents is any integer between 1 and the upper limit. When the number of halogen substituents is greater than 1, it can be the substitution of the same or different halogens.

[0198] In this article, "oxo" or "oxygen group" refers to double-bonded oxygen (=O). "O" and "oxygen group" are used interchangeably in this article.

[0199] In this specification, for the purpose of brevity, the groups defined above may be used in combination to describe groups not defined in the definition section herein, wherein each group individually has the meaning as defined above. For example, "alkylaminoalkyl" means that the hydrogen in the alkyl group is replaced by an amino group, and the hydrogen in the amino group is replaced by an alkyl group; for example, methylaminoethyl means -C2H5-NH-CH3, or "cyanophenyl" means that the hydrogen on the phenyl group is replaced by a cyano group, methoxyamino means -NHOMe, and dimethylamino means -NMe2.

[0200] The term "connecting unit" refers to a divalent molecular chain that links two parts of a compound. This molecular chain can be composed of atoms, groups, bonds, etc., and the hydrogen atoms on the molecular chain can optionally be replaced by other groups.

[0201] The connecting lines drawn from a ring system indicate that one end of the bond can be connected to any suitable ring atom in the ring system that the bond passes through. Passing through one ring indicates a connection at any optional position within that ring; passing through multiple rings indicates a connection at any optional position within those multiple rings. For example, The structure includes any of the following structures:

[0202] It should be understood that the terms related to the groups defined above are not mutually exclusive.

[0203] The terms "optional substitution" or "optionally substituted by one or more substituents" mean that the hydrogen on the referred group is not substituted or that one or more substituted sites of the referred group are independently substituted by groups selected from the following groups: halogens (e.g., -F, -Cl, -Br, -I), amino groups, C groups. 1-10 Alkyl groups (e.g., methyl, ethyl, n-propyl, isopropyl, cyclopropyl, tert-butyl), haloalkyl groups (e.g., trifluoromethyl), C 2-10 Alkenyl (e.g., propenyl, allyl), C 2-10 Alkyne, cycloalkyl, heterocyclic, aryl (e.g., phenyl), heteroaryl, alkoxy (e.g., methoxy, ethoxy), thiol, cyano, nitro, hydroxy, carboxyl, ester, ketone, aldehyde, sulfonic acid, phosphate, silyl, and any suitable group described herein.

[0204] The term "treatment" for a subject's disease or "treatment" for a subject with or suspected of having a disease refers to administering medication to the subject, such as one or more agents, to reduce or prevent the worsening of at least one symptom of the disease. Therefore, in one implementation, "treatment" specifically refers to delaying progression, accelerating remission, inducing remission, increasing remission, accelerating recovery, increasing the efficacy of alternative therapies, or reducing resistance to alternative therapies, or combinations thereof. For example, reducing or maintaining the number or volume of detectable tumors in a treated patient population relative to an untreated control group, or reducing their invasiveness and metastatic potential, or reducing their resistance to known therapies or chemotherapy, etc.

[0205] In some embodiments, a "therapeutic effective amount" means that applying a sufficient amount of the compound disclosed in this invention will, to some extent, alleviate, reduce, or cure one or more symptoms of the treated disease or condition. In some embodiments, the result is a reduction and / or mitigation of the signs, symptoms, or causes of the disease, or any other desired alteration of the biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising the compound disclosed in this application required to provide a clinically significant reduction in disease symptoms.

[0206] The term "cancer" can include cancers resulting from genetic mutations. Examples of such cancers include, but are not limited to, breast cancer; cancers associated with Lee-Flaumeni syndrome, such as childhood sarcoma, leukemia, and brain cancer; cancers associated with Lynch syndrome, such as colon cancer, bile duct cancer, brain cancer, endometrial cancer, kidney cancer, ovarian cancer, pancreatic cancer, small intestine cancer, stomach cancer, and ureteral cancer; lung cancer; melanoma; prostate cancer; retinoblastoma; thyroid cancer; and uterine cancer. Furthermore, cancer can be the result of acquired mutations, such as mutations caused by diet, environment, and / or lifestyle, or somatic mutations. Examples of such cancers may include, but are not limited to, adrenal carcinoma, adrenocortical carcinoma, bladder cancer, brain cancer, primary brain cancer, glioma, glioblastoma, breast cancer, cervical cancer, colon cancer (non-limiting examples include colorectal cancers such as colonic adenocarcinoma and colon cancer), endometrial cancer, epidermal cancer, esophageal cancer, gallbladder cancer, genitourinary tract cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer (non-limiting examples include adenocarcinoma, small cell lung cancer and non-small cell lung cancer), lymphoma (non-limiting examples include B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma), melanoma, malignant melanoma, malignant carcinoid, malignant pancreatic insulinoma, myeloma, multiple myeloma, ovarian cancer, pancreatic cancer (e.g., exocrine pancreatic cancer), prostate cancer, renal cell carcinoma, skin cancer, and, for example, squamous cell carcinoma, gastric cancer, testicular cancer, thyroid cancer, and thyroid follicle cancer, among others. Follicular carcinoma, Wilms' tumor, choriocarcinoma, fungal infection, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, pilocellular lymphoma, Burkholderia lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, promyelocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia, fibrosarcoma, habdomyosarcoma, astrocytoma, neuroblastoma, rhabdomyosarcoma, schwannoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, seminoma, teratoma, osteosarcoma, xenodermal melanoma, keratokeratoma, and retinoblastoma. The cancers envisioned in this invention, particularly MSI cancers, are characterized in a preferred embodiment by high microsatellite instability. In this application, MSI cancer and MSI tumor have the same meaning.

[0207] The term "subject" refers to an animal, such as a mammal, like a human or veterinary animal, including but not limited to primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In some implementations, the subject is a human.

[0208] The term "pharmaceutical composition" refers to a mixture of one or more compounds described in this invention or their pharmaceutically acceptable salts, stereoisomers, isotopic compounds or prodrugs, with other components, wherein the other components contain physiologically / pharmaceuticalally acceptable carriers and / or excipients.

[0209] The term "medicinal salt" refers to (i) the salt formed by an acidic functional group (e.g., -COOH) present in the compounds provided by this invention and a suitable inorganic or organic cation (base), including but not limited to, alkali metal salts such as sodium, potassium, and lithium salts; alkaline earth metal salts such as calcium and magnesium salts; other metal salts such as aluminum, iron, zinc, copper, nickel, and cobalt salts; inorganic base salts such as ammonium salts; and organic base salts such as tert-octylamine, dibenzylamine, morpholine, glucosamine, phenylglycine alkyl ester, ethylenediamine, N-methylglucosamine, guanidine, diethylamine, triethylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, chloroprocaine, procaine, diethanolamine, N-benzyl-phenylethylamine, piperazine, tetramethylamine, and tris(hydroxymethyl)aminomethane. And (ii) the salts formed by the basic functional group (e.g., -NH2) present in the compounds provided by this invention and suitable inorganic or organic anions (acids), including but not limited to, hydrohalides such as hydrofluoric acid, hydrochloride, hydrobromide, hydroiodide, etc.; inorganic acid salts such as nitrates, perchlorates, sulfates, phosphates, etc.; lower alkyl sulfonates such as methanesulfonates, trifluoromethanesulfonates, ethanesulfonates, etc.; aryl sulfonates such as benzenesulfonates, p-benzenesulfonates, etc.; organic acid salts such as acetates, malates, fumarates, succinates, citrates, tartrates, oxalates, maleates, etc.; amino acid salts such as glycine salts, trimethylglycine salts, arginine salts, ornithine salts, glutamate salts, aspartate salts, etc. See Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002).

[0210] The compounds or salts of this invention may exist as enantiomers and diastereomers resulting from axial chirality or chiral centers. All such stereoisomers are part of this invention. Independent stereoisomers of the compounds of this invention may not coexist with other isomers (e.g., possessing special activity as a pure or substantially pure optical isomer), or may be mixtures, such as racemates, or mixtures formed with all other stereoisomers or a portion thereof.

[0211] The term "isotope derivative" refers to an isotopically labeled compound. Examples of isotopes that can be listed as compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, such as 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F, and 36Cl, respectively. In some embodiments, heavier isotopes, such as deuterium, i.e., 2H or D, are substituted, which may be preferred in certain cases due to their good metabolic stability, which is advantageous in some therapies, such as increasing the half-life or reducing the dosage in vivo. Isotopically labeled compounds can be prepared using general methods by replacing non-isotopic reagents with readily available isotopically labeled reagents, according to the schemes disclosed in the examples.

[0212] The term "prodrug" refers to a derivative of the compound that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide the compound of the present invention. Prodrugs become active compounds only after undergoing the reaction under biological conditions, or they do not have or only have low activity in their unreacted forms. Prodrugs can generally be prepared using well-known methods, such as those described in Burger's Medicinal Chemistry and Drug Discovery (1995) 172-178, 949-982 (Manfred E. Wolff, 5th edition).

[0213] The pharmaceutical compositions of the present invention may contain pharmaceutically acceptable excipients, including but not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycerol, sorbic acid, potassium sorbate, a mixture of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, beeswax, lanolin, etc.

[0214] "Pharmaceutically acceptable carriers" refer to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here refers to the ability of the components in the composition to interact with and be mixed with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0215] "Pharmaceutically acceptable excipients" refer to substances that contribute to the formulation and / or administration and / or absorption of an 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, aqueous 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.

[0216] In some embodiments, the pharmaceutical compositions of the present invention may be in solid or liquid form.

[0217] Drugs containing active ingredients can be in suitable oral dosage forms, 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 drugs can be prepared according to known process methods of the drug 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. Tablets contain the active ingredient mixed with non-toxic, pharmaceutically acceptable excipients suitable for tablet production. Examples of these 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.

[0218] The pharmaceutical compositions of the present invention can be prepared in various forms according to different routes of administration. For example, the pharmaceutical compositions can be administered in any of the following ways: orally, by spray inhalation, rectal administration, nasal administration, buccal administration, vaginal administration, topical administration, and extra-intestinal administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intrasternal, and intracranial injection or infusion, or via an external implantation device. Oral or intravenous administration is preferred.

[0219] The compounds of this invention may optionally be used in combination with one or more other active ingredients, the amounts and proportions of which may be adjusted by those skilled in the art according to specific symptoms, patient conditions, and clinical needs. Combined use does not mean that the therapy or treatment agent must be administered simultaneously and / or formulated for joint delivery, although such delivery methods are within the scope of this invention. The combined therapeutic agent may be administered simultaneously, before, or after one or more other additional therapies or treatment agents. The therapeutic agent or treatment regimen may be administered in any order.

[0220] "Optional" or "optionally" means that the event or situation described below may but does not have to happen, and the description includes the circumstances under which the event or situation may or may not happen.

[0221] Some of the exemplary compounds in Formula I can be produced by one or more of the following synthetic methods. The substituent identifiers R2, R3, R4, R5, R6, R7, R8 and R9, as well as K2 to K7 and X, in the methods have the meanings defined above for compounds of Formula I and Formula II, and x or y can be selected from Cl, Br or I.

[0222] Synthesis Method 1

[0223]

[0224] Synthesis Method 2

[0225]

[0226] Synthesis Method 3

[0227]

[0228] Synthesis Method 4

[0229]

[0230] Synthesis Method 5

[0231]

[0232] Synthesis Method 6

[0233]

[0234] Synthesis Method 7

[0235]

[0236] Synthesis Method 8

[0237]

[0238] Synthesis Method 9

[0239]

[0240] Synthesis Method 10

[0241]

[0242] Synthesis Method 11

[0243]

[0244] The present invention will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed by the present invention.

[0245] Example

[0246] Unless otherwise specified, the compounds used in the embodiments of this application are obtained through conventional commercial purchases. The abbreviations used in this application and their meanings are shown in Table 2.

[0247] Table 2. Abbreviations used in this application and their meanings

[0248]

[0249] Preparation of intermediates

[0250] 1. Preparation of intermediate A (Int-A)

[0251]

[0252] Step 1: Dissolve 4-chlorobenzoyl hydrazide (50 mg, 293 μmol) in MeOH (0.5 mL), then add ethyl isothiocyanate (26 mg, 293 μmol, 26 μL) and react at 65 °C for 15 minutes. After the reaction is complete, evaporate the solvent to obtain Int-A-1 (75 mg, 99% yield). No purification is required; it can be used directly in the next step.

[0253] Step 2: Add Int-A-1 (70 mg, 272 μmol) to the reaction flask and add 10% NaOH solution (2 mL). React at 85 °C for 16 hours. After the reaction is complete, adjust the pH to 2 with 10% dilute hydrochloric acid, filter, collect the filter cake, and dry to obtain Int-A (62 mg, yield 95%), which is a white powder.

[0254] 2. Preparation of intermediate B (Int-B)

[0255]

[0256] The preparation method of Int-B is the same as that of Int-A, except that 4-chlorobenzoylhydrazine and methyl isothiocyanate are used as raw materials, and the overall yield of the two-step reaction is 87%.

[0257] 3. Preparation of intermediate C (Int-C)

[0258]

[0259] Procedure: 1-(6-bromopyridin-3-yl)acetone (100 mg, 0.5 mmol), pinacol ester of 3,6-dihydro-2H-pyran-4-boronate (210 mg, 1 mmol), tetrakis(triphenylphosphine)palladium (58 mg, 50 μmol), and potassium carbonate (208 mg, 1.5 mmol) were dissolved in 1,4-dioxane (2 mL). After purging with nitrogen six times, the reaction was carried out at 90 °C for 16 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted three times with EA, and the organic phase was dried and purified by column chromatography to obtain Int-C (100 mg, 98% yield).

[0260] 4. Preparation of intermediate D (Int-D)

[0261]

[0262] Step 1: 4-Chlorobenzoylhydrazide (2 g, 11.7 mmol) and N,N-dimethylformamide dimethyl acetal (1.4 g, 11.7 mmol, 1.6 mL) were dissolved in ACN (45 mL). After reacting at 50 °C for 1 hour, methylamine hydrochloride (752 mg, 11.1 mmol) and acetic acid (12 mL) were added sequentially. The temperature was raised to 120 °C and the reaction was continued for 16 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and Int-D-1 (1.22 g, yield 53%) was purified by column chromatography.

[0263] Step 2: Int-D-1 (500 mg, 2.6 mmol) was dissolved in formaldehyde solution (15.5 mL, 37% in H2O, 206 mmol). The reaction was carried out overnight at 90 °C. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and Int-D-2 (550 mg, 95% yield) was obtained by column chromatography.

[0264] Step 3: Int-D-2 (200 mg, 894 μmol) was dissolved in DCM (3 mL). Carbon tetrabromide (415 mg, 1.25 mmol) and triphenylphosphine (328 mg, 1.25 mmol) were added sequentially at 0 °C. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted with DCM, dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by column chromatography to obtain Int-D-3 (209 mg, yield 81%).

[0265] Step 4: Dissolve Int-D-3 (209 mg, 729 μmol) in EtOH (2 mL), and add NH4OH solution (2 mL, 50% inH2O). React at 30 °C for 24 hours. After the reaction is complete as monitored by TLC, desolvate under reduced pressure and purify by column chromatography to obtain Int-D (155 mg, yield 95%).

[0266] 5. Preparation of intermediate E (Int-E)

[0267]

[0268] Step 1: 2,4-Dibromopyridine (1 g, 4.22 mmol), 3,4-dimethoxyphenylboronic acid (769 mg, 4.22 mmol), tetrakis(triphenylphosphine)palladium (488 mg, 422 μmol), and sodium carbonate (895 mg, 8.44 mmol) were dissolved in a mixed solvent of toluene (12 mL), EtOH (6 mL), and water (3 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 100 °C. After the reaction was monitored by TLC, water was added to quench the reaction, followed by extraction with EA, solvent removal under reduced pressure, and purification by column chromatography to obtain Int-E-1 (970 mg, yield 78%).

[0269] Step 2: Int-E-1 (500 mg, 1.7 mmol), B(pin)2 (518 mg, 2 mmol), tetrakis(triphenylphosphine)palladium (197 mg, 170 μmol), and potassium acetate (501 mg, 5.1 mmol) were dissolved in 1,4-dioxane (10 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 90 °C. After the reaction was completed by TLC monitoring, the reaction was quenched with water, then extracted with EA, dissolved under reduced pressure, and purified by column chromatography to obtain Int-E (502 mg, yield 86%).

[0270] 6. Preparation of intermediate F (Int-F)

[0271]

[0272] Step 1: Dissolve iminoaldehyde (460 mg, 4.8 mmol) in ACN (10 mL), and add dropwise bromo-3,4-dimethoxyacetophenone solution (1 g in 10 mL ACN, 3.8 mmol). After reacting at 75 °C for 6 hours, filter while hot. Wash the filter residue three times with ACN, collect the filtrate, and remove the solid under reduced pressure. Reflux in 5 N hydrochloric acid (10 mL) for 1 hour, then add isopropanol for azeotropic removal. The resulting solid is reacted further in isopropanol (20 mL) at room temperature for 0.5 hours. Filter, wash the filter cake with diethyl ether, and dry to obtain Int-F-1. The crude product is not purified further and is used directly in the next reaction.

[0273] Step 2: Dissolve Int-F-1 (250 mg, 1.3 mmol) in MeOH (5 mL). Cool to 0 °C and add sodium borohydride (97 mg, 2.6 mmol). React at room temperature for 1 hour, then quench the reaction with saturated ammonium chloride solution (2 mL). After desolvation under reduced pressure, add 1 N NaOH solution (5 mL), then extract with DCM. Dry the organic phase and desolvate under reduced pressure to obtain Int-F (160 mg). The overall yield of the two-step reaction is 60%.

[0274] 7. Preparation of intermediate G (Int-G)

[0275]

[0276] Step 1: 2-Nitro-4,5-dimethoxy-2'-bromoacetophenone (587 mg, 1.93 mmol), potassium iodide (64 mg, 386 μmol), and potassium acetate (227 mg, 2.32 mmol) were dissolved in DMF (8 mL). The reaction was carried out at 30 °C for 2 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted three times with DCM, and purified by column chromatography to obtain Int-G-1 (545 mg, 99% yield).

[0277] Step 2: Int-G-1 (545 mg, 1.92 mmol) was dissolved in a mixed solvent of EA (5 mL) and MeOH (5 mL). After cooling to 0 °C, stannous chloride dihydrate (1.3 g, 5.77 mmol) was added. The reaction was carried out at 30 °C for 12 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with 10% sodium bicarbonate solution, and the solid precipitated and filtered. The filtrate was extracted three times with EA, the organic phase was dried, and the solvent was removed under reduced pressure. The solution was purified by column chromatography to obtain Int-G-2 (361 mg, yield 74%).

[0278] Step 3: Dissolve Int-G-2 (361 mg, 1.44 mmol) in THF (5 mL), then add lithium hydroxide solution (2 mL, 1 min H2O). React at 30 °C for 1 hour. After the reaction is complete as monitored by TLC, quench the reaction with DCM, adjust the pH to 4 with 10% dilute hydrochloric acid, extract three times with DCM, dry the organic phase, remove the solvent under reduced pressure, and purify by column chromatography to obtain Int-G-3 (241 mg, yield 80%).

[0279] Step 4: Dissolve Int-G-3 (70 mg, 335 μmol) in DCM (1 mL). After cooling to 0 °C, add triphenylphosphine (122 mg, 468 μmol) and carbon tetrabromide (155 mg, 468 μmol). React at 0 °C for 10 minutes. After the reaction is complete as monitored by TLC, purify Int-G (60 mg, yield 66%) by column chromatography as soon as possible. Store at low temperature and protected from light for subsequent reactions.

[0280] 8. Preparation of intermediates H and I (Int-H and Int-I)

[0281]

[0282] Procedure: 2'-Amino-4',5'-dimethoxyacetophenone (500 mg, 2.56 mmol) was dissolved in acetic acid (5 mL), followed by the addition of liquid bromine (614 mg, 3.84 mmol, 197 μL). The reaction was carried out at 30 °C for 18 hours. After the reaction was complete, sodium thiosulfate solution was added to quench the reaction, and the mixture was extracted with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain Int-H (240 mg, 34% yield) and Int-I (140 mg, 15% yield). How to separate them?

[0283] 9. Preparation of intermediate J (Int-J)

[0284]

[0285] Procedure: 4-Aminopyridazine (285 mg, 3 mmol) and 1,1-thiocarbonylbis-2(1H)-pyridone (765 mg, 3.3 mmol) were dissolved in ultradry DCM (30 mL). After purging with nitrogen six times, the reaction was carried out at room temperature for 6 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water and extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain Int-J (200 mg, yield 48%).

[0286] 10. Synthesis of the intermediate (Int-K)

[0287]

[0288] The preparation method of Int-K is the same as that of Int-J, except that 4-aminopyrazole and 1,1-thiocarbonylbis-2(1H)-pyridone are used as raw materials, and the reaction yield is 37%.

[0289] 11. Synthesis of intermediate L (Int-L)

[0290]

[0291] The preparation method of Int-L is the same as that of Int-G, except that 2-nitro-4-methoxy-5-fluoro-2'-bromoacetophenone is used as the starting material, and the overall yield of the four-step reaction is 27%.

[0292] Example 1

[0293] Preparation of 2-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)-1-(3,4-dihydroxyphenyl)ethyl-1-one (compound 1)

[0294]

[0295] Steps: Synthesis of Compound 1

[0296] Int-A (30 mg, 125 μmol) was dissolved in DMF (0.5 mL), and 3,4-dihydroxy-2'-chloroacetophenone (24 mg, 125 μmol) and potassium carbonate (52 mg, 375 μmol) were added. The reaction was carried out at room temperature for 16 hours. After the reaction was complete, the solvent was removed under reduced pressure, and the solution was purified by silica gel column chromatography to give compound 1 (36 mg, yield 72%).

[0297] 1 H NMR (400MHz, DMSO-d6) δ10.03(s,1H),9.42(s,1H),7.62-7.69(m,4H),7.43(dd,J=8.3,2.2Hz,1H),7.37(d, J=2.2Hz,1H),6.83(d,J=8.3Hz,1H),4.88(s,2H),4.02(q,J=6.9Hz,2H),1.22(t,J=7.3Hz,3H).ESI-MS[M+H] + :390.1.

[0298] Examples 2-26:

[0299] Referring to the synthesis method of Example 1, the corresponding compounds, namely compounds 2 to 26, were prepared using the raw materials arylformylhydrazine and isothiocyanate listed in Table 3.

[0300] Table 3: Compounds from Examples 2-26

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308]

[0309] Example 27

[0310] Preparation of 2-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)-1-(4-hydroxy-3-methoxyphenyl)ethyl-1-one (compound 27)

[0311]

[0312] Step 1: Synthesis of Compound 27-1

[0313] 3-Methoxy-4-hydroxyacetophenone (100 mg, 602 μmol) was dissolved in a mixture of chloroform (2 mL) and ethyl acetate (1 mL), and cuprous bromide (134 mg, 602 μmol) was added. The reaction was carried out at 60 °C for 12 hours. After the reaction was completed by TLC monitoring, the mixture was filtered, dissolved under reduced pressure, and purified by column chromatography to give compound 27-1 (100 mg, yield 67%).

[0314] Step 2: Synthesis of Compound 27

[0315] Int-A (20 mg, 83 μmol) was dissolved in DMF (0.5 mL), and 27-1 (31 mg, 125 μmol) and potassium carbonate (24 mg, 170 μmol) were added. The reaction was carried out at room temperature for 4 hours. After the reaction was complete, the compound 27 (10 mg, yield 29%) was purified by silica gel column chromatography.

[0316] 1H NMR (400MHz, DMSO-d6) δ10.16(s,1H),7.61-7.69(m,4H),7.59(dd,J=8.3,2.0Hz,1H),7.49(d,J=1.9Hz,1H) ,6.89(d,J=8.3Hz,1H),4.90(s,2H),4.03(q,J=7.2Hz,2H),3.83(s,3H),1.21(t,J=7.3Hz,3H).ESI-MS[M+H] + 404.1.

[0317] Examples 28-48:

[0318] Referring to the synthesis method of Example 27, using Int-A or Int-B as raw materials, the corresponding compounds, namely compounds 28 to 48, were prepared using the acetophenone raw materials in Table 4.

[0319] Table 4: Compounds from Examples 28-48

[0320]

[0321]

[0322]

[0323]

[0324]

[0325] Example 49

[0326] Preparation of 1-(2-amino-1H-benzo[d]imidazol-6-yl)-2-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)ethyl-1-one (compound 49)

[0327]

[0328] Step 1: Synthesis of Compound 49-1

[0329] 2-Aminobenzimidazole (50 mg, 376 mmol) and bromoacetic acid (53 mg, 376 mmol, 27 μL) were dissolved in polyphosphoric acid (1 mL) and reacted overnight at 115 °C. After quenching the reaction with saturated sodium bicarbonate solution, the mixture was extracted with DCM, and the organic phase was dried over anhydrous sodium sulfate. The crude product 49-1 obtained after desolvation under reduced pressure was used directly in the next reaction without further purification.

[0330] Step 2: Synthesis of Compound 49

[0331] Int-A (8 mg, 32 μmol) was dissolved in DMF (0.5 mL), and 49-1 (8 mg, 32 μmol) and potassium carbonate (7 mg, 50 μmol) were added. The reaction was carried out at room temperature for 4 hours. After the reaction was completed, the product was purified by silica gel column chromatography to obtain compound 49 (5 mg, overall yield of the two-step reaction was 5%).

[0332] 1 H NMR (400MHz, Methanol-d4) δ7.58-7.65(m,4H),7.47-7.50(m,2H),7.22-7.24(m,2H),4.17(q,J=7.3Hz,2H),1.33(t,J=7.3Hz,3H).ESI-MS[M+H] + :413.0.

[0333] Example 50

[0334] Preparation of 5-(2-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)acetyl)-N,2-dihydroxybenzamide (compound 50)

[0335]

[0336] Step 1: Synthesis of Compound 50-1

[0337] Int-B (41 mg, 183 μmol), methyl 2-hydroxy-5-(2-bromoacetyl)benzoate (50 mg, 183 μmol), and potassium carbonate (38 mg, 275 μmol) were dissolved in DMF (2 mL). The reaction was carried out overnight at 30 °C. After the reaction was completed by TLC, a saturated ammonium chloride solution was added to quench the reaction, followed by extraction with DCM. The organic phase was dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by column chromatography to give 50-1 (67 mg, 87% yield).

[0338] Step 2: Synthesis of Compound 50

[0339] Hydroxylamine hydrochloride (292 mg, 4.2 mmol) was dissolved in 3.5 mL of water, and sodium hydroxide solution (400 mg in 1.5 mL H₂O, 10 mmol) was added dropwise. After stirring for 5 minutes, a hydroxylamine solution was obtained and set aside. Compound 50-1 (10 mg, 24 mmol) and sodium hydroxide (2 mg, 50 μmol) were dissolved in a mixed solvent of THF (0.5 mL) and MeOH (0.5 mL), and then 0.1 mL of the above hydroxylamine solution was added. The reaction was carried out overnight at room temperature. The reaction solution was diluted with water, and the pH was adjusted to 7-8 with 1 M hydrochloric acid. Extraction was performed using DCM, and the organic phase was dried over anhydrous sodium sulfate and dissolved under reduced pressure. The compound 50 (1 mg, 9% yield) was purified using semi-preparative high-performance liquid chromatography.

[0340] 1 H NMR (400MHz, Methanol-d4) δ8.41(s,1H),8.07(d,J=8.7Hz,1H),7.59-7.72(m,6H),7.03(d,J=8.2Hz,1H),4.81(q,J=7.2Hz,2H),3.72(s,3H).ESI-MS[M+H] + :419.1.

[0341] Example 51

[0342] Preparation of 1-(3,4-dihydroxyphenyl)-2-((5-(4-hexylphenyl)-4-propyl-4H-1,2,4-triazol-3-yl)thio)ethyl-1-one (compound 51)

[0343]

[0344] Step 1: Synthesis of Compound 51-1

[0345] Propyl isothiocyanate (5.06 g, 50 mmol, 5.2 mL) and formyl hydrazide (3.3 g, 55 mmol) were dissolved in THF (200 mL). The reaction was carried out overnight at room temperature, and a large amount of solid precipitated. The solid was filtered. After drying the filter cake, it was redissolved in DMF (80 mL), and potassium carbonate solution (6.91 g in 20 mL H2O, 50 mmol) was added. The reaction was carried out overnight at 50 °C. After the reaction was completed by TLC monitoring, the reaction solution was poured into ice water. The pH was adjusted to 5-6 by adding 2 M dilute hydrochloric acid, and the mixture was filtered. The filter cake was washed with water and dried to obtain crude product 51-1 (5.87 g, yield 82%).

[0346] Step 2: Synthesis of compound 51-2

[0347] 51-1 (716 mg, 5 mmol), ethyl 3-iodopropionate (1.37 g, 6 mmol, 800 mL), and potassium carbonate (2.08 g, 15 mmol) were dissolved in DMF (20 mL). The reaction was carried out at room temperature for 2 hours. After the reaction was completed by TLC, the reaction was quenched with ice water and extracted three times with DCM. After drying and solvent removal of the organic phase, the solution was purified by silica gel column chromatography to obtain 51-2 (902 mg, yield 74%).

[0348] Step 3: Synthesis of compound 51-3

[0349] 51-2 (730 mg, 3 mmol) and NBS (641 mg, 3.6 mmol) were dissolved in ACN (10 mL). The reaction was carried out at room temperature for 12 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water and extracted three times with DCM. The organic phase was washed successively with 1% sodium thiosulfate solution and saturated brine, dried and desolventized, and purified by silica gel column chromatography to obtain 51-3 (552 mg, yield 57%).

[0350] Step 4: Synthesis of compound 51-4

[0351] 51-3 (484 mg, 1.5 mmol) was dissolved in MeOH (10 mL), and then lithium hydroxide solution (190 mg in 0.5 mL H₂O, 4.5 mmol) was added. The reaction was allowed to proceed at room temperature for 15 minutes. After TLC monitoring showed complete reaction, the reaction was quenched with water, and the pH was adjusted to 5-6 with 2 M dilute hydrochloric acid. The mixture was then extracted three times with DCM. The organic phase was washed with saturated brine, dried, and purified by silica gel column chromatography to obtain 51-4 (267 mg, 80% yield).

[0352] Step 5: Synthesis of compound 51-5

[0353] 51-4 (350 mg, 1.6 mmol), 2-chloro-1-(2,2-dimethylbenzo[D][1,3]dioxolane-5-yl)acetone (381 mg, 1.7 mmol), and potassium carbonate (930 mg, 6.7 mmol) were dissolved in DMF (60 mL). The reaction was carried out overnight at room temperature. After the reaction was completed by TLC, the mixture was filtered. The filtrate was diluted with EA (200 mL) and H2O (150 mL). After separation, the organic phase was washed three times with water (100 mL × 3), dried over anhydrous sodium sulfate, and purified by silica gel column chromatography under reduced pressure to obtain 51-5 (330 mg, 50% yield).

[0354] Step 6: Synthesis of Compounds 51-6

[0355] 51-5 (60 mg, 145 μmol), 4-n-hexylphenylboronic acid (45 mg, 218 μmol), potassium carbonate (41 mg, 291 μmol), and dichlorobis(triphenylphosphine)palladium (11 mg, 15 μmol) were dissolved in a mixed solvent of DME (2 mL) and H₂O (0.5 mL). After reacting in a microwave reactor at 150 °C for 30 min, a saturated NaHCO₃ solution was added, and the mixture was extracted with DCM. The organic phase was dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by preparative TLC to obtain 51-6 (14 mg, 19% yield).

[0356] Step 7: Synthesis of Compound 51

[0357] Compound 51-6 (7 mg, 14 μmol) was dissolved in TFA (1 mL) and reacted at room temperature for 4 hours. After desolvation under reduced pressure, compound 51 (2 mg, 33% yield) was purified by semi-preparative high performance liquid chromatography.

[0358] 1 H NMR (400MHz, Methanol-d4) δ7.41-7.53(m,6H),6.84(d,J=8.2Hz,1H),4.82(s,2H),4.05(t,J=7.6Hz,2H),2.72(t,J=7.6 Hz,2H),2.00-2.04(m,2H),1.59-1.70(m,2H),1.30-1.34(m,6H),0.88-0.92(m,3H),0.88(t,J=7.4Hz,3H).ESI-MS[M+H] + :454.2.

[0359] Examples 52-55:

[0360] Referring to the synthesis method of Example 51, the corresponding compounds, namely compounds 52 to 55, were prepared using the isothiocyanate raw materials in Table 5 and commercially available phenylboronic acid raw materials.

[0361] Table 5: Compounds from Examples 52-55

[0362]

[0363]

[0364] Example 56

[0365] Preparation of 3-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)-1-(3,4-dihydroxyphenyl)prop-1-one (compound 56)

[0366]

[0367] Step 1: Synthesis of Compound 56-1

[0368] Int-A (30 mg, 125 μmol) and 3-chloro-3',4'-dimethoxyphenylacetone (29 mg, 125 μmol) were dissolved in DCM (1 mL), followed by the addition of TEA (38 mg, 375 μmol, 53 μL) and DMAP (3 mg, 25 μmol). The reaction was carried out at 30 °C for 18 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted with DCM, and the organic phase was dried over anhydrous sodium sulfate and purified by column chromatography to obtain 56-1 (10 mg, 18% yield).

[0369] Step 2: Synthesis of Compound 56

[0370] 56-1 (10 mg, 23 μmol) was dissolved in DCM (1 mL), and BBr3 (58 mg, 230 μmol, 23 μL) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the compound 56 (4 mg, yield 40%) was purified by reverse semi-preparative high performance liquid chromatography.

[0371] 1 H NMR (400MHz, DMSO-d6) δ9.90(s,1H),9.37(s,1H),7.67(d,J=8.4Hz,2H),7.62(d,J=8.5Hz,2H),7.36(d,J=10.4Hz,2H), 6.80(d,J=8.1Hz,1H),3.94(q,J=7.1Hz,2H),3.44-3.46(m,2H),2.65-2.70(m,2H),1.16(t,J=7.2Hz,3H).ESI-MS[M+H] + :404.1.

[0372] Example 57

[0373] Preparation of 4-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)-1-(3,4-dihydroxyphenyl)but-1-one (compound 57)

[0374]

[0375] Step 1: Synthesis of Compound 57-1

[0376] In a reaction flask, phthalic acid (100 mg, 724 μmol, 93 μL) and 4-bromobutyryl chloride (134 mg, 724 μmol, 84 μL) were dissolved in DCM (5 mL). AlCl3 (116 mg, 870 μmol) was added in portions at 0 °C. The reaction was carried out at room temperature for 1 hour. TLC monitoring was performed. After the reaction was complete, the temperature was lowered to 0 °C, and the reaction was quenched with ice water. The mixture was extracted with DCM, washed once with water, dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by column chromatography to obtain 57-1 (164 mg, 78% yield).

[0377] Step 2: Synthesis of Compound 57-2

[0378] 57-1 (50 mg, 209 μmol), TEA (43 mg, 418 μmol, 59 μL), and DMAP (3 mg, 21 μmol) were dissolved in DCM (1 mL). The reaction was carried out at 30 °C for 18 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted with DCM, and the organic phase was dried over anhydrous sodium sulfate and purified by column chromatography to obtain 57-2 (75 mg, 80% yield).

[0379] Step 3: Synthesis of Compound 57

[0380] 57-2 (30 mg, 67 μmol) was dissolved in DCM (1 mL), and BBr3 (169 mg, 230 μmol, 23 μL) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until it was complete. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 57 (4 mg, yield 40%).

[0381] 1 H NMR (400MHz, DMSO-d6) δ9.85(s,1H),9.34(s,1H),7.68(d,J=8.6Hz,2H),7.62(d,J=8.6Hz,2H),7.32-7.45(m,2H),6.80(d,J=8.0H z,1H),3.98(q,J=7.2Hz,2H),3.26(t,J=7.2Hz,2H),3.05(t,J=7.1Hz,2H),1.98-2.03(m,2H),1.18(t,J=7.2Hz,3H).ESI-MS[M+H] + :418.1.

[0382] Examples 58-59:

[0383] Referring to the synthesis method of Example 57, the corresponding compounds, namely compounds 58 to 59, were prepared using the acyl chloride raw materials in Table 6.

[0384] Table 6: Compounds from Examples 58-59

[0385]

[0386]

[0387] Example 60

[0388] Preparation of 4-(2-((5-(4-chlorophenyl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)ethyl)phenyl-1,2-diol (compound 60)

[0389]

[0390] Steps: Synthesis of Compound 60

[0391] Int-A (25 mg, 104 μmol) was dissolved in acetone (1 mL), and 4-(2-bromoethyl)benzene-1,2-diol (27 mg, 125 μmol) and potassium carbonate (18 mg, 125 μmol) were added. The reaction was carried out at room temperature for 16 hours. After the reaction was complete, the product was purified by silica gel column chromatography to give compound 60 (28 mg, 71% yield).

[0392] 1 H NMR (400MHz, DMSO-d6) δ8.79(s,1H),8.71(s,1H),7.68(d,J=8.6Hz,2H),7.62(d,J=8.6Hz,2H),6.63(d,J=7.7Hz,2H) ,6.41-6.55(m,1H),3.87-4.01(m,2H),3.36-3.40(m,2H),2.84(t,J=7.5Hz,2H),1.16(t,J=7.2Hz,3H).ESI-MS[M+H] + :376.1.

[0393] Example 61

[0394] Preparation of 4-(4-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)methyl)pyridin-2-yl)phenyl-1,2-diol (compound 61)

[0395]

[0396] Step 1: Synthesis of Compound 61-1

[0397] Methyl 6-bromonicotinic acid (1 g, 4.63 mmol), 3,4-dimethoxyphenylboronic acid (1.01 g, 5.55 mmol), and tetrakis(triphenylphosphine)palladium (268 mg, 231 μmol) were dissolved in 1,4-dioxane (20 mL). After purging the gas with nitrogen six times, 2.0 M sodium carbonate solution (5 mL) was added. The reaction was carried out overnight at 80 °C. After TLC showed that the reaction was complete, water was added to quench the reaction, and the solution was extracted with EA, dissolved under reduced pressure, and purified by column chromatography to obtain 61-1 (730 mg, yield 57%).

[0398] Step 2: Synthesis of Compound 61-2

[0399] Dissolve 61-1 (730 mg, 2.67 mmol) in THF (15 mL). After cooling to 0 °C, add lithium aluminum hydride (136 mg, 4 mmol). React at room temperature for 15 minutes. After TLC shows the reaction is complete, quench the reaction with water (2 mL), then add 1.0 M NaOH solution (2 mL). After filtration through diatomaceous earth, extract with EA, and desolvate under reduced pressure to obtain crude 61-2, which is used directly in the next step without further purification.

[0400] Step 3: Synthesis of compound 61-3

[0401] Int-B (50 mg, 221 μmol), 61-2 (66 mg, 266 μmol), and triphenylphosphine (88 mg, 332 μmol) were dissolved in THF (2 mL). After purging the gas with nitrogen six times, the mixture was cooled to 0 °C, and then diisopropyl azodicarbonate (77 mg, 378 μmol, 75 μL) was added. The reaction was allowed to proceed overnight at room temperature. After the reaction was monitored by TLC, water was added to quench the reaction, and the mixture was extracted with EA, dissolved under reduced pressure, and purified by preparative TLC to obtain 61-3 (36 mg, 35% yield).

[0402] Step 4: Synthesis of Compound 61

[0403] 61-3 (12 mg, 26 μmol) was dissolved in DCM (1 mL), and BBr3 (130 μL, 2.0 M in DCM, 260 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 61 (9 mg, yield 79%).

[0404] 1H NMR (400MHz, Methanol-d4) δ8.57(d,J=6.1Hz,1H),8.07(d,J=1.7Hz,1H),7.82(dd,J=1.7,6.1Hz,1H) ,7.54-7.66(m,4H),7.25-7.36(m,2H),6.98(d,J=8.3Hz,1H),4.60(s,2H),3.65(s,3H).ESI-MS[M+H] + :425.1.

[0405] Example 62

[0406] Preparation of 2-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)-N-(3,4-dihydroxyphenylethyl)acetamide (compound 62)

[0407]

[0408] Step 1: Synthesis of Compound 62-1

[0409] 3,4-Dimethoxyphenethylamine (200 mg, 1.1 mmol) was dissolved in DCM (4 mL), and TEA (223 mg, 2.2 mmol, 308 μL) was added. After cooling to 0 °C, chloroacetyl chloride (187 mg, 1.66 mmol, 132 μL) was added dropwise. The reaction was carried out at room temperature for 2 hours. After the reaction was monitored by TLC, saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted with DCM, dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by column chromatography to obtain 62-1 (260 mg, 91% yield).

[0410] Step 2: Synthesis of Compound 62-2

[0411] Int-B (44 mg, 194 μmol), 62-1 (50 mg, 194 μmol), and potassium carbonate (54 mg, 388 μmol) were dissolved in DMF (2 mL). The reaction was carried out overnight at 30 °C. After the reaction was monitored by TLC, saturated ammonium chloride was added to quench the reaction. The mixture was extracted with DCM, dried over anhydrous sodium sulfate, and dissolved under reduced pressure. The solution was purified by preparative TLC to obtain 62-2 (72 mg, 83% yield).

[0412] Step 3: Synthesis of Compound 62

[0413] 62-2 (12 mg, 27 μmol) was dissolved in DCM (1 mL), and BBr3 (135 μL, 2.0 M in DCM, 270 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 62 (8 mg, yield 69%).

[0414] 1 H NMR (400MHz, CDCl3) δ7.34 (d, J=8.6Hz, 2H), 7.17-7.28 (m, 2H), 6.21-6.31 (m, 2H), 6.11 (dd, J=2 .1,8.0Hz,1H),3.50(s,2H),3.31(s,3H),2.96-3.02(m,2H),2.25(t,J=7.1Hz,2H).ESI-MS[M+H] + 419.1.

[0415] Example 63

[0416] Preparation of 2-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)-N-(3,4-dihydroxyphenyl)ethyl-1-one (compound 63)

[0417]

[0418] Steps: Synthesis of Compound 63

[0419] Int-C (35 mg, 156 μmol) was dissolved in DMF (0.5 mL), and 3,4-dihydroxy-2'-chloroacetophenone (35 mg, 187 μmol) and potassium carbonate (33 mg, 234 μmol) were added. The reaction was carried out overnight at 40 °C. After the reaction was complete, the solvent was removed under reduced pressure, and the solution was purified by silica gel column chromatography to give compound 63 (42 mg, 72% yield).

[0420] 1 H NMR(400MHz,DMSO-d6)δ10.10(s,1H),9.44(s,1H),7.52-7.68(m,5H),7.32 -7.43(m,2H),6.84(d,J=8.2Hz,1H),4.80(s,2H),3.69(s,3H).ESI-MS[M+H] + :375.1.

[0421] Example 64

[0422] Preparation of 2-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)quinazolin-6,7-diol (compound 64)

[0423]

[0424] Step 1: Synthesis of Compound 64-1

[0425] Int-B (15 mg, 66 μmol) and 2-chloro-6,7-dimethoxyquinazoline (15 mg, 66 μmol) were dissolved in n-butanol (2 mL), and concentrated hydrochloric acid (5 μL) was added. The mixture was refluxed overnight, and the solvent was removed under reduced pressure. The solution was purified by preparative TLC to obtain 64-1 (5 mg, yield 17%).

[0426] Step 2: Synthesis of Compound 64

[0427] 64-1 (5 mg, 11 μmol) was dissolved in DCM (1 mL), and BBr3 (55 μL, 2.0 M in DCM, 110 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the compound 64 (1.5 mg, yield 35%) was purified by reverse semi-preparative high performance liquid chromatography.

[0428] 1 H NMR(400MHz, CDCl3)δ8.91(s,1H),7.77-7.86(m,2H),7.60-7.72(m,2H),7.19(s,1H),7.01(s,1H),3.77(s,3H).ESI-MS[M+H] + :386.1.

[0429] Example 65

[0430] Preparation of 1-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)methyl)-3,4-dihydroisoquinoline-6,7-diol (compound 65)

[0431]

[0432] Step 1: Synthesis of Compound 65-1

[0433] 62-2 (20 mg, 45 μmol) and phosphorus oxychloride (137 mg, 895 μmol, 84 μL) were dissolved in toluene (2 mL). The mixture was refluxed for 2 hours, then dissolved under reduced pressure and purified by preparative TLC to obtain 65-1 (6 mg, 31% yield).

[0434] Step 2: Synthesis of Compound 65

[0435] 65-1 (6 mg, 14 μmol) was dissolved in DCM (1 mL), and BBr3 (70 μL, 2.0 M in DCM, 140 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 65 (3 mg, yield 53%).

[0436] 1 H NMR (400MHz, CDCl3) δ7.56-7.68(m,4H),7.27(d,J=5.6Hz,1H),6.82(d,J=3.9Hz,1H) ,3.81(td,J=8.2,2.9Hz,2H),3.65(d,J=6.7Hz,3H),2.94-3.06(m,2H).ESI-MS[M+H] + :401.1.

[0437] Example 66

[0438] Preparation of 2-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)amino)-1-(3,4-dihydroxyphenyl)ethyl-1-one (compound 66)

[0439]

[0440] Steps: Synthesis of Compound 66

[0441] Int-D (10 mg, 45 μmol), 3,4-dihydroxy-2'-chloroacetophenone (11 mg, 58 μmol), and potassium carbonate (19 mg, 135 μmol) were dissolved in DMF (1 mL). The reaction was carried out at 30 °C for 3 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and compound 66 (7 mg, 40% yield) was purified by preparative TLC.

[0442] 1 H NMR(600MHz,Methanol-d4)δ7.71(d,J=8.4Hz,2H),7.66(d,J=8.5Hz,2H),7.41-7.5 1(m,2H),6.92(d,J=8.1Hz,1H),4.89(s,2H),4.66(s,2H),3.79(s,3H).ESI-MS[M+H] + :373.1.

[0443] Example 67

[0444] Preparation of N-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)-2-(3,4-dihydroxyphenyl)acetamide (compound 67)

[0445]

[0446] Steps: Synthesis of Compound 67

[0447] Int-D (15 mg, 67 μmol), 3,4-dihydroxyphenylacetic acid (17 mg, 101 μmol), EDCI (16 mg, 81 μmol), HOBt (11 mg, 81 μmol), and DIPEA (26 mg, 202 μmol, 35 μL) were dissolved in DMF (0.8 mL). The reaction was carried out at 30 °C for 18 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and compound 67 (17 mg, 68% yield) was purified by preparative TLC.

[0448] 1 H NMR(600MHz, Methanol-d4)δ7.59-7.69(m,4H),6.76(d,J=2.1Hz,1H),6.72(d,J=8.0Hz ,1H),6.64(dd,J=8.1,2.1Hz,1H),4.64(s,2H),3.56(s,3H),3.43(s,2H).ESI-MS[M+H] + 373.1.

[0449] Example 68

[0450] Preparation of N-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)-3,4-dihydroxybenzenesulfonamide (compound 68)

[0451]

[0452] Step 1: Synthesis of Compound 68-1

[0453] Int-D (14 mg, 63 μmol) was dissolved in DCM (1 mL). After cooling to 0 °C, 3,4-dimethoxybenzenesulfonyl chloride (15 mg, 63 μmol) and pyridine (25 mg, 317 μmol, 26 μL) were added sequentially. The reaction was carried out at room temperature for 16 hours. After the reaction was completed as monitored by TLC, the solvent was removed under reduced pressure, and the solution was purified by preparative TLC to obtain 68-1 (17 mg, 63% yield).

[0454] Step 2: Synthesis of Compound 68

[0455] 68-1 (10 mg, 23 μmol) was dissolved in DCM (1 mL), and BBr3 (115 μL, 2.0 M in DCM, 230 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the compound 68 (7 mg, 74% yield) was purified by reverse semi-preparative high performance liquid chromatography.

[0456] 1 H NMR (400MHz, DMSO-d6) δ9.90 (s, 1H), 9.65 (s, 1H), 8.05 (d, J = 6.2Hz, 1H), 7.60-7.68 (m, 4H), 7.18 (s ,1H),7.10(d,J=8.3Hz,1H),6.85(d,J=8.3Hz,1H),4.15(d,J=5.7Hz,2H),3.58(s,3H).ESI-MS[M+H] + :395.1.

[0457] Example 69

[0458] Preparation of 2-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)(methyl)amino)-1-(3,4-dihydroxyphenyl)ethyl-1-one (compound 69)

[0459]

[0460] Step 1: Synthesis of Compound 69-1

[0461] Int-D-3 (50 mg, 174 μmol), methylamine hydrochloride (36 mg, 522 μmol), and DIPEA (113 mg, 870 μmol, 150 μL) were dissolved in MeOH (1 mL). The reaction was carried out at 30 °C for 16 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 69-1 (27 mg, 65% yield).

[0462] Step 2: Synthesis of Compound 69-2

[0463] 69-1 (30 mg, 126 μmol), bromo-3,4-dimethoxyacetophenone (40 mg, 152 μmol), and potassium carbonate (53 mg, 380 μmol) were dissolved in DMF (0.5 mL). The reaction was carried out at 50 °C for 3 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted three times with DCM, dried the organic phase, removed the solvent under reduced pressure, and purified by column chromatography to obtain 69-2 (40 mg, yield 76%).

[0464] Step 3: Synthesis of Compound 69

[0465] 69-2 (30 mg, 72 μmol) was dissolved in DCM (1 mL), and BBr3 (180 μL, 2.0 M in DCM, 360 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 69 (18 mg, yield 66%).

[0466] 1 H NMR (600MHz, DMSO-d6) δ10.29(s,1H),9.57(s,1H),7.75(d,J=8.7Hz,2H),7.68(d,J=8.7Hz,2H),7.43 -7.30(m,2H),6.90(d,J=8.2Hz,1H),4.96(s,2H),4.60(s,2H),3.76(s,3H),2.97(s,3H).ESI-MS[M+H] + :387.1.

[0467] Example 70

[0468] Preparation of 2-chloro-4-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)amino)quinazoline-6,7-diol (compound 70)

[0469]

[0470] Step 1: Synthesis of Compound 70-1

[0471] Int-D (22 mg, 0.1 mmol), 2,4-dichloro-6,7-dimethoxyquinazoline (20 mg, 77 μmol), and DIPEA (20 mg, 154 μmol, 27 μL) were dissolved in MeOH (2 mL). The reaction was carried out overnight at 30 °C. After the reaction was completed by TLC, the solvent was removed under reduced pressure and purified by column chromatography to give 70-1 (6 mg, yield 17%).

[0472] Step 2: Synthesis of Compound 70

[0473] 70-1 (5 mg, 11 μmol) was dissolved in DCM (1 mL), and BBr3 (28 μL, 2.0 M in DCM, 55 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 70 (2 mg, yield 44%).

[0474] 1H NMR (600MHz, Methanol-d4) δ7.72(d,J=8.3Hz,2H),7.65(d,J=8.2Hz,2H),7.44-7.51(m,1H),7.01(s,1H),5.07(s,2H),3.92(s,3H).ESI-MS[M+H] + :417.1.

[0475] Example 71

[0476] Preparation of 2-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)amino)quinazoline-6,7-diol (compound 71)

[0477]

[0478] Compound 71 was prepared from Int-D and 2-chloro-6,7-dimethoxyquinazoline using the synthetic method of Example 64, with an overall yield of 5% in the two-step reaction.

[0479] 1 H NMR(400MHz, Methanol-d4)δ9.07(s,1H),7.70(d,J=8.3Hz,2H),7.63(d,J=8.5Hz,2H),7.30(s,1H),7.05(s,1H),3.82(s,3H).ESI-MS[M+H] + 383.1.

[0480] Example 72

[0481] Preparation of 4-(4-(5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)pyridin-2-yl)phenyl-1,2-diol (compound 72)

[0482]

[0483] Step 1: Synthesis of Compound 72-1

[0484] Int-D-1 (200 mg, 1 mmol) was dissolved in DCM (5 mL). After cooling to 0 °C, NBS (203 mg, 1.1 mmol) was added. The reaction was allowed to proceed overnight at room temperature. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction. The mixture was then extracted with DCM, dissolved under reduced pressure, and purified by column chromatography to obtain 72-1 (100 mg, 35% yield).

[0485] Step 2: Synthesis of compound 72-2

[0486] 72-1 (50 mg, 183 μmol), Int-E (94 mg, 275 μmol), tetrakis(triphenylphosphine)palladium (22 mg, 18 μmol), and potassium carbonate (51 mg, 367 μmol) were dissolved in a mixed solvent of 1,4-dioxane (1.4 mL) and water (0.6 mL). The reaction was carried out in a microwave reactor at 110 °C for 1.5 hours. The reaction was then quenched with saturated sodium bicarbonate solution, followed by extraction with DCM. The organic phase was dried over anhydrous sodium sulfate, dissolved under reduced pressure, and purified by preparative TLC to obtain 72-2 (12 mg, 16% yield).

[0487] Step 3: Synthesis of Compound 72

[0488] 72-2 (12 mg, 29 μmol) was dissolved in DCM (1 mL), and BBr3 (145 μL, 2.0 M in DCM, 290 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 72 (8 mg, 70% yield).

[0489] 1 H NMR(400MHz, Methanol-d4)δ9.01(d,J=2.2Hz,1H),8.44(dd,J=8.6,2.2Hz,1H),8.17(d,J=8.5Hz,1H),7.82-7.88(m,2H) ,7.68-7.75(m,2H),7.59(d,J=2.3Hz,1H),7.51(dd,J=8.4,2.3Hz,1H),6.98(d,J=8.3Hz,1H),3.89(s,3H).ESI-MS[M+H] + 379.1.

[0490] Example 73

[0491] Preparation of 4-(4-((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)methyl)pyridin-2-yl)phenyl-1,2-diol (compound 73)

[0492]

[0493] Step 1: Synthesis of Compound 73-1

[0494] Int-D-3 (168 mg, 586 μmol), Int-E (200 mg, 586 μmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (65 mg, 88 μmol), and cesium carbonate (573 mg, 1.76 mmol) were dissolved in a mixture of DME (3 mL) and water (0.5 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 110 °C. After the reaction was completed by TLC monitoring, water was added to quench the reaction, followed by extraction with EA, solvent removal under reduced pressure, and purification by preparative TLC to obtain 73-1 (11 mg, yield 4%).

[0495] Step 2: Synthesis of Compound 73

[0496] 72-2 (11 mg, 26 μmol) was dissolved in DCM (1 mL), and BBr3 (130 μL, 2.0 M in DCM, 260 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 73 (1 mg, yield 9%).

[0497] 1 H NMR (400MHz, Methanol-d4) δ8.62(s,1H),8.07(s,1H),7.96(d,J=8.2Hz,1H),7.70(d,J=8.5Hz,2H),7.59-7.6 5(m,2H),7.42(d,J=2.3Hz,1H),7.34(dd,J=8.3,2.3Hz,1H),6.95(d,J=8.3Hz,1H),4.43(s,2H).ESI-MS[M+H] + :393.1.

[0498] Example 74

[0499] Preparation of 1-(3,4-dihydroxyphenyl)-2-(1H-indazol-1-yl)ethyl-1-one (compound 74)

[0500]

[0501] Steps: Synthesis of Compound 74

[0502] 1H-indazole (40 mg, 338 μmol) was dissolved in DMF (0.5 mL). After cooling to 0 °C, sodium hydride (68 mg, 1.69 mmol, 60% mineral oil) was added. After stirring for 30 minutes, 3,4-dihydroxy-2'-chloroacetophenone (70 mg in 0.5 mL DMF, 372 μmol) was added. The reaction was carried out at 30 °C for 2 hours. After the reaction was completed by TLC monitoring, methanol was added to quench the reaction, and the solvent was removed under reduced pressure. The solution was purified by column chromatography to give compound 74 (22 mg, 23% yield).

[0503] 1 H NMR (600MHz, DMSO-d6) δ10.05(s,1H),9.45(s,1H),8.11(s,1H),7.79(d,J=8.1Hz,1H),7.55(d,J=8.4Hz,2H),7 .39–7.46(m,1H),7.35(t,J=7.6Hz,1H),7.15(d,J=8.4Hz,1H),6.89(d,J=8.2Hz,1H),5.99(s,2H).ESI-MS[M+H] + :269.1.

[0504] Example 75

[0505] Preparation of 4-(2-(1H-indazol-1-yl)acetyl)-1,2-phenylenediacetate (compound 75)

[0506]

[0507] Steps: Synthesis of Compound 75

[0508] Compound 74 (30 mg, 111 μmol), TEA (34 mg, 335 μmol, 47 μL), and DMAP (2 mg, 16 μmol) were dissolved in DCM (1 mL). After cooling to 0 °C, acetyl chloride (27 mg, 335 μmol, 21 μL) was slowly added. After the reaction was monitored by TLC until complete, the solvent was removed under reduced pressure, and compound 75 (9 mg, 21% yield) was purified by preparative TLC.

[0509] 1H NMR (600MHz, CDCl3) δ8.08(d,J=1.0Hz,1H),7.96(dd,J=8.5,2.1Hz,1H),7.89(d,J=2.1Hz,1H),7.74-7.78(m,1H ),7.34-7.41(m,2H),7.26-7.30(m,1H),7.15-7.20(m,1H),5.77(s,2H),2.33(s,3H),2.32(s,3H).ESI-MS[M+H] + :353.1.

[0510] Examples 76-89:

[0511] Referring to the synthesis method of Example 74, the corresponding compounds, namely compounds 76 to 89, were prepared using the nitrogen-containing aromatic ring raw materials in Table 7.

[0512] Table 7: Compounds from Examples 75-89

[0513]

[0514]

[0515]

[0516] Example 90

[0517] Preparation of 1-(3,4-dihydroxyphenyl)-2-(quinazolin-2-ylamino)ethyl-1-one (compound 90)

[0518]

[0519] Step 1: Synthesis of Compound 90-1

[0520] 2-Chloroquinazoline (60 mg, 364 μmol) and Int-F (72 mg, 364 μmol) were dissolved in n-butanol (2 mL), and concentrated hydrochloric acid (10 μL) was added. The mixture was heated under reflux overnight, dissolved under reduced pressure, and purified by preparative TLC to give 90-1 (10 mg, 8% yield).

[0521] Step 2: Synthesis of Compound 90-2

[0522] 90-1 (10 mg, 30 μmol) was dissolved in ultradry DCM (1 mL), the gas was purged with nitrogen six times, and after cooling to 0 °C, DMP (20 mg, 46 μmol) was added. The reaction was carried out at room temperature for 0.5 h, the reaction was quenched with 10% sodium sulfite solution, and then saturated sodium bicarbonate solution was added. After stirring thoroughly for 5 min, the mixture was extracted with DCM, dissolved under reduced pressure, and purified by preparative TLC to obtain 90-2 (4.4 mg, yield 44%).

[0523] Step 3: Synthesis of Compound 90

[0524] 90-2 (4.4 mg, 13 μmol) was dissolved in DCM (1 mL), and BBr3 (65 μL, 2.0 M in DCM, 130 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 90 (2 mg, yield 49%).

[0525] 1 H NMR (600MHz, Methanol-d4) δ9.32(brs,1H),8.00(brs,1H),7.91(brs,1H),7.65(d,J=8.5Hz,1H),7.55(dd,J=8.3,2.0H z,1H),7.44-7.49(m,2H),7.29(t,J=7.6Hz,1H),7.20(d,J=6.2Hz,1H),6.90(d,J=7.9Hz,2H),5.05(s,2H).ESI-MS[M+H] + :296.1.

[0526] Examples 91-92:

[0527] Referring to the synthesis method of Example 90, the corresponding compounds, namely compounds 91 to 92, were prepared using the chloroaromatic raw materials listed in Table 8.

[0528] Table 8: Compounds from Examples 91-92

[0529]

[0530]

[0531] Example 93

[0532] Preparation of 1-(2-(3,4-dihydroxyphenyl)-2-oxoethyl)pyridine-2(1H)-one (compound 93)

[0533]

[0534] Compound 93 was prepared using the synthetic method of Example 74, with a yield of 21%, from 3,4-dihydroxy-2'-chloroacetophenone and 2-hydroxypyridine.

[0535] 1 H NMR (400MHz, DMSO-d6) δ10.01(s,1H),9.45(s,1H),7.60(dd,J=6.7,2.1Hz,1H),7.46(ddt,J=10.4,6.6,2.2Hz,2H),7 .39(d,J=2.1Hz,1H),6.87(d,J=8.3Hz,1H),6.37-6.45(m,1H),6.24(td,J=6.7,1.4Hz,1H),5.35(s,2H).ESI-MS[M+H] + 246.1.

[0536] Example 94

[0537] Preparation of 1-(3,4-dihydroxyphenyl)-2-(6-morpholino-1H-indazol-1-yl)ethyl-1-one (compound 94)

[0538]

[0539] Steps: Synthesis of Compound 94

[0540] Compound 76 (30 mg, 86 μmol), morpholine (16 mg, 173 μmol, 16 μL), tris(dibenzylacetone)dipalladium (9 mg, 10 μmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (6 mg, 10 μmol), and sodium tert-butoxide (17 mg, 173 mmol) were dissolved in anhydrous toluene (0.5 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 110 °C. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by reverse semi-preparative high-performance liquid chromatography to give compound 94 (4 mg, yield 13%).

[0541] 1 H NMR (600MHz, DMSO-d6) δ10.00(s,1H),9.43(s,1H),7.89(s,1H),7.59(d,J=8.8Hz,1H),7.53(dd,J=8.3,2.1Hz,1H),7.41( d,J=2.1Hz,1H),6.92-7.01(m,2H),6.89(d,J=8.3Hz,1H),5.88(s,2H),3.72-3.77(m,4H),3.13-3.17(m,4H).ESI-MS[M+H] + :354.1.

[0542] Examples 95-96:

[0543] Referring to the synthesis method of Example 94, the corresponding compounds, namely compounds 95 to 96, were prepared using the starting amines in Table 9.

[0544] Table 9: Compounds from Examples 95-96

[0545]

[0546] Example 97

[0547] Preparation of 1-(3,4-dihydroxyphenyl)-2-(6-phenyl-1H-indazol-1-yl)ethyl-1-one (compound 97)

[0548]

[0549] Steps: Preparation of Compound 97

[0550] Compound 76 (20 mg, 47 μmol), phenylboronic acid (9 mg, 71 μmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (4 mg, 5 μmol), and potassium carbonate (20 mg, 142 μmol) were dissolved in 1,4-dioxane (0.5 mL) and water (0.5 mL). After purging the gas with nitrogen six times, the reaction was carried out at 90 °C for 16 hours. After the reaction was monitored by TLC, the reaction was quenched with water, extracted three times with EA, the organic phases were combined, dried and desolventized, and purified by reverse semi-preparative high performance liquid chromatography to give compound 97 (5 mg, yield 30%).

[0551] 1 H NMR (600MHz, DMSO-d6) δ9.84(s,1H),9.53(s,1H),8.12(d,J=0.6Hz,1H),7.93(s,1H),7.86(d,J=8.4Hz,1H),7.67-7.80(m,2H),7.56(d d,J=8.3,2.1Hz,1H),7.45-7.52(m,3H),7.43(d,J=2.1Hz,1H),7.37(t,J=7.4Hz,1H),6.89(d,J=8.2Hz,1H),6.07(s,2H).ESI-MS[M+H] + :345.1.

[0552] Examples 98-123

[0553] Referring to the synthesis method of Example 97, compounds 98 to 123 were prepared by using compound 76 or compound 83 as raw materials and boric acid (ester) listed in Table 10.

[0554] Table 10: Compounds from Examples 98 to 123

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565] Example 124

[0566] Preparation of 1-(2-(3,4-dihydroxyphenyl)-2-oxoethyl)-5-(thiophen-3-yl)pyrimidin-2(1H)-one (compound 124)

[0567]

[0568] Step 1: Synthesis of Compound 124-1

[0569] 3,4-Dihydroxy-2'-chloroacetophenone (45 mg, 240 μmol) and potassium iodide (40 mg, 240 μmol) were dissolved in acetone (2 mL). After reacting at 50 °C for 20 min, 5-bromo-2-hydroxypyrimidine (28 mg, 160 μmol) and potassium carbonate (66 mg, 480 μmol) were added sequentially. The reaction was continued at 50 °C for 2 h. After the reaction was completed by TLC monitoring, the mixture was filtered, the filtrate was quenched with methanol, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 124-1 (13 mg, yield 22%).

[0570] Step 2: Synthesis of Compound 124

[0571] Compound 124-1 (32 mg, 100 μmol), 3-thiopheneboronic acid (13 mg, 100 μmol), tetrakis(triphenylphosphine)palladium (12 mg, 10 μmol), and potassium carbonate (42 mg, 300 μmol) were dissolved in a mixed solvent of 1,4-dioxane (2.5 mL) and water (0.75 mL). After reacting for 1 hour at 100 °C in a microwave reactor, the pH was adjusted to 6 with dilute hydrochloric acid, and the solvent was removed under reduced pressure. The solution was purified by reverse semi-preparative high-performance liquid chromatography to give compound 124 (4 mg, yield 11%).

[0572] 1 H NMR (400MHz, Methanol-d4) δ9.07 (s, 1H), 8.47 (s, 1H), 7.70 (dd, J = 3.0, 1.3Hz, 1H), 7.56-7.62 (m, 2H),7.51(d,J=2.2Hz,1H),7.43(d,J=1.4Hz,1H),6.92(d,J=8.3Hz,1H),5.52(s,2H).ESI-MS[M+H] + :329.1.

[0573] Examples 125-135

[0574] Referring to the synthesis method of Example 124, the corresponding compounds, namely compounds 125 to 135, were prepared using the boric acid (ester) raw material listed in Table 11.

[0575] Table 11: Compounds from Examples 125-135

[0576]

[0577]

[0578]

[0579]

[0580] Example 136

[0581] Preparation of 3-(((5-(4-chlorophenyl)-1H-imidazol-2-yl)thio)methyl)benzo[c]isoxazole-5,6-diol (compound 136)

[0582]

[0583] Step 1: Synthesis of Compound 136-1

[0584] 2-Amino-4'-chloroacetophenone hydrochloride (300 mg, 1.46 mmol) and potassium thiocyanate (156 mg, 1.60 mmol) were dissolved in acetic acid (1.5 mL). The mixture was heated under reflux for 1 hour, and the color of the reaction solution gradually changed from orange to colorless. The reaction was quenched with water after cooling to 0 °C, producing a large amount of white precipitate. The precipitate was filtered, and the filter cake was dried to obtain crude product 136-1 (273 mg, yield 89%). No further purification was required; it was used directly in the next reaction.

[0585] Step 2: Synthesis of Compound 136-2

[0586] 136-1 (50 mg, 237 μmol) and Int-G (65 mg, 237 μmol) were dissolved in DCM (0.5 mL), and then TEA (72 mg, 712 μmol, 100 μL) was added. The reaction was carried out at 30 °C for 3 hours. After the reaction was monitored by TLC until complete, the reaction was quenched with water, and then extracted three times with DCM. After drying the organic phase, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 136-2 (50 mg, yield 52%).

[0587] Step 3: Synthesis of Compound 136

[0588] 136-2 (15 mg, 37 μmol) was dissolved in DCM (1 mL), and BBr3 (185 μL, 2.0 M in DCM, 370 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 136 (4 mg, yield 27%).

[0589] 1 H NMR (600MHz, DMSO-d6) δ12.54(s,1H),10.15(s,1H),7.78(d,J=7.9Hz,2H),7.73(s,1H),7.47 (d,J=7.5Hz,1H),7.41(d,J=7.9Hz,2H),6.63(s,1H),6.54(s,1H),4.76(s,2H).ESI-MS[M+H] + :374.0.

[0590] Example 137

[0591] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)benzo[c]isoxazole-5,6-diol (compound 137)

[0592]

[0593] Step 1: Synthesis of Compound 137-1

[0594] 136-2 (25 mg, 62 μmol), bromoacetonitrile (38 mg, 311 μmol, 22 μL), potassium iodide (11 mg, 62 μmol), and potassium carbonate (26 mg, 186 μmol) were dissolved in DMF (0.5 mL). The reaction was carried out at 50 °C for 3 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 137-1 (20 mg, yield 72%).

[0595] Step 2: Synthesis of Compound 137

[0596] 137-1 (15 mg, 34 μmol) was dissolved in DCM (1 mL), and BBr3 (170 μL, 2.0 M in DCM, 340 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 137 (6 mg, yield 39%).

[0597] 1 H NMR(600MHz,DMSO-d6)δ10.38(s,1H),9.64(s,1H),7.91(s,1H),7.75(d,J=8.4Hz,2H), 7.43(d,J=8.4Hz,2H),6.63(s,1H),6.55(s,1H),5.20(s,2H),4.79(s,2H).ESI-MS[M+H] + :412.0.

[0598] Examples 138-140

[0599] Referring to the synthesis method of Example 137, the corresponding compounds, namely compounds 138 to 140, were prepared using the brominated raw materials listed in Table 12.

[0600] Table 12: Compounds from Examples 138-140

[0601]

[0602] Example 141

[0603] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)benzo[c]isoxazole-5,6-diol (compound 141)

[0604]

[0605] Step 1: Synthesis of Compound 141-1

[0606] 2-Amino-4'-chloroacetophenone hydrochloride (500 mg, 2.43 mmol) and methyl isothiocyanate (895 mg, 12.2 mmol, 835 μL) were dissolved in pyridine (12.5 mL). The reaction was carried out at 30 °C for 2 h, then increased to 90 °C and reacted for another 1.5 h. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 141-1 (428 mg, yield 78%).

[0607] Step 2: Synthesis of Compound 141-2

[0608] 141-1 (225 mg, 1 mmol), Int-G (272 mg, 1 mmol), and potassium carbonate (415 mg, 3 mmol) were dissolved in DMF (3 mL). The reaction was carried out at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure and purified by column chromatography to obtain 141-2 (170 mg, yield 41%).

[0609] Step 3: Synthesis of Compound 141

[0610] 141-2 (20 mg, 48 μmol) was dissolved in DCM (1 mL), and BBr3 (240 μL, 2.0 M in DCM, 480 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 141 (7 mg, yield 35%).

[0611] 1 H NMR (400MHz, DMSO-d6) δ10.38(s,1H),9.74(s,1H),7.55(d,J=8.4Hz,2H),7.42(d ,J=8.4Hz,3H),6.55(s,1H),6.36(s,1H),4.71(s,2H),3.31(s,3H).ESI-MS[M+H] + :388.0.

[0612] Examples 142-165

[0613] Referring to the synthesis method of Example 141, the corresponding compounds, namely compounds 142 to 165, were prepared using α-aminoaryl ketone and isothiocyanate listed in Table 13.

[0614] Table 13: Compounds from Examples 142-165

[0615]

[0616]

[0617]

[0618]

[0619]

[0620]

[0621]

[0622] Example 165

[0623] Preparation of 3-(2-(((5,6-dihydroxybenzo[c]isoxazol-3-yl)methyl)thio)-1-methyl-1H-imidazol-5-yl)benzoic acid (compound 165)

[0624]

[0625] Steps: Synthesis of Compound 165

[0626] Compound 161 (8 mg, 20 μmol) was dissolved in THF (1 mL), and then 1 M lithium hydroxide aqueous solution was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was monitored by TLC, the reaction was quenched with DCM, and the pH was adjusted to 4 with 10% dilute hydrochloric acid. The mixture was extracted three times with DCM, and the organic phase was dried and desolvated under reduced pressure. The solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 165 (5 mg, yield 67%).

[0627] 1 H NMR(600MHz, Methanol-d4)δ8.19(dt,J=7.8,1.4Hz,1H),8.06(t,J=1.8Hz,1H),7.74(s,1H),7.66(t,J=7 .7Hz,1H),7.61(dt,J=7.7,1.5Hz,1H),6.61(s,1H),6.33(s,1H),4.75(s,2H),3.41(s,3H).ESI-MS[M+H] + :398.1.

[0628] Example 166

[0629] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)-6,7-dihydro-[1,4]dioxin[2',3':4,5]benzo[1,2-c]isoxazole (compound 166)

[0630]

[0631] Step 1: Synthesis of Compound 166-1

[0632] 2'-Bromo-1-(7-amino-2,3-dihydro-1,4-benzodioxin-6-yl) ethyl ketone (604 mg, 2 mmol), potassium iodide (67 mg, 400 μmol), and potassium acetate (236 mg, 2.4 mmol) were dissolved in DMF (8 mL). The reaction was carried out at 30 °C for 2 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water, extracted three times with DCM, and purified by column chromatography to obtain 166-1 (534 mg, 95% yield).

[0633] Step 2: Synthesis of Compound 166-2

[0634] 166-1 (534 mg, 1.9 mmol) was dissolved in a mixed solvent of EA (5 mL) and MeOH (5 mL). After cooling to 0 °C, stannous chloride dihydrate (1.3 g, 5.7 mmol) was added. The reaction was carried out at 30 °C for 12 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with 10% sodium bicarbonate solution, and the solid precipitated and filtered. The filtrate was extracted three times with EA, the organic phase was dried, and the solvent was removed under reduced pressure. The solution was purified by column chromatography to give compound 166-2 (360 mg, 76% yield).

[0635] Step 3: Synthesis of Compound 166-3

[0636] 166-2 (360 mg, 1.44 mmol) was dissolved in THF (5 mL), and then lithium hydroxide solution (2 mL, 1 M in H2O) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was monitored by TLC, the reaction was quenched with DCM, and the pH was adjusted to 4 with 10% dilute hydrochloric acid. The mixture was extracted three times with DCM, and the organic phase was dried and desolvated under reduced pressure. The solution was purified by column chromatography to obtain 166-3 (224 mg, yield 75%).

[0637] Step 4: Synthesis of Compound 166-4

[0638] 166-3 (63 mg, 300 μmol) was dissolved in DCM (1 mL). After cooling to 0 °C, triphenylphosphine (110 mg, 420 μmol) and carbon tetrabromide (140 mg, 420 μmol) were added. The reaction was carried out at 0 °C for 10 min. After the reaction was completed by TLC monitoring, 166-4 (47 mg, 58% yield) was purified by column chromatography as soon as possible.

[0639] Step 5: Synthesis of Compound 166

[0640] Compound 166-4 (47 mg, 174 μmol) and 141-1 (40 mg, 174 μmol) were dissolved in DCM (0.5 mL), and then TEA (53 mg, 522 μmol, 73 μL) was added. The reaction was carried out at 30 °C for 3 hours. After the reaction was monitored by TLC until complete, the reaction was quenched with water, and then extracted three times with DCM. After drying the organic phase, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to give compound 166 (33 mg, yield 47%).

[0641] 1 H NMR (600MHz, CDCl3) δ7.36-7.40(m,2H),7.23(s,1H),7.11-7.16(m,2H),6.84(d,J=0.6Hz,1H ),6.29(s,1H),4.55(s,2H),4.23-4.27(m,2H),4.14-4.18(m,2H),3.05(s,3H).ESI-MS[M+H] + :414.1.

[0642] Example 167

[0643] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)benzo[c]isoxazole-5,6-diester (compound 167)

[0644]

[0645] Steps: Synthesis of Compound 167

[0646] Compound 154 (6 mg, 16 μmol), DMAP (2 mg, 1.6 μmol), and TEA (5 mg, 49 μmol, 7 μL) were dissolved in DCM (1 mL), and propionyl chloride (5 mg, 50 μmol, 5 μL) was slowly added dropwise. After the reaction was monitored by TLC until complete, the solvent was removed under reduced pressure, and the solution was purified by reverse semi-preparative high-performance liquid chromatography to give compound 167 (3 mg, yield 38%).

[0647] 1 H NMR (600MHz, Methanol-d4) δ7.61 (s, 1H), 7.46-7.51 (m, 3H), 7.31 (t, J = 8.7Hz, 2H), 7.25 (s, 1H), 4.91 (s, 2H), 3.56 (s, 3H), 2. 66(qd,J=7.6,1.1Hz,2H),2.60(qd,J=7.6,1.1Hz,2H),1.26(td,J=7.7,1.1Hz,3H),1.20(td,J=7.5,1.0Hz,3H).ESI-MS[M+H]+ :484.1.

[0648] Example 168

[0649] Preparation of 3-(((5-(5-chloropyridin-2-yl)-1-methyl-1H-imidazol-2-yl)thio)methyl)benzo[c]isoxazole-5,6-diol (compound 168)

[0650]

[0651] Step 1: Synthesis of Compound 168-1

[0652] 1-(5-chloropyridin-2-yl)acetone (300 mg, 1.93 mmol) was dissolved in hydrobromic acid solution (4 mL, 33% in HOAc), and then Br2 (339 mg, 2.12 mmol, 109 μL) was added. The reaction was carried out at 30 °C for 3 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with diethyl ether. The mixture was filtered, the filter cake was washed with diethyl ether, collected, and dried to give crude product 168-1 (557 mg, yield 91%).

[0653] Step 2: Synthesis of Compound 168-2

[0654] 168-1 (300 mg, 951 μmol) and sodium azide (186 mg, 2.85 mmol) were dissolved in a mixture of acetone (2 mL) and water (3 mL). The reaction was carried out at 30 °C for 15 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water and extracted three times with EA. After drying the organic phase, the solvent was removed under reduced pressure to obtain crude 168-2 (180 mg, 96% yield).

[0655] Step 3: Synthesis of Compound 168-3

[0656] 168-2 (100 mg, 508 μmol) and stannous chloride dihydrate (345 mg, 1.53 mmol) were dissolved in MeOH (1 mL), and then concentrated hydrochloric acid (0.2 mL) was added. The reaction was carried out at 30 °C for 12 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 168-3 (100 mg, yield 94%).

[0657] Step 4: Synthesis of Compound 168-4

[0658] 168-3 (100 mg, 586 μmol) and methyl isothiocyanate (215 mg, 2.93 mmol, 200 μL) were dissolved in pyridine (1 mL). The reaction was carried out at 30 °C for 2 hours, and then the temperature was increased to 90 °C for another 1.5 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 168-4 (97 mg, yield 73%).

[0659] Step 5: Synthesis of Compound 168-5

[0660] 168-4 (97 mg, 428 μmol), Int-G (117 mg, 428 μmol), and potassium carbonate (178 mg, 1.28 mmol) were dissolved in DMF (1 mL). The reaction was carried out at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 168-5 (82 mg, yield 46%).

[0661] Step 6: Synthesis of Compound 168

[0662] 168-5 (12 mg, 29 μmol) was dissolved in DCM (1 mL), and BBr3 (145 μL, 2.0 M in DCM, 290 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the product was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 168 (9 mg, yield 77%).

[0663] 1 H NMR (600MHz, DMSO-d6) δ10.35(s,1H),9.71(s,1H),8.66(d,J=2.3Hz,1H),8.01(dd,J=8.6,2.5H z,1H),7.79(d,J=8.7Hz,2H),6.54(s,1H),6.51(s,1H),4.77(s,2H),3.69(s,3H).ESI-MS[M+H] + :389.0.

[0664] Examples 169-175

[0665] Referring to the synthesis method of Example 168, the corresponding compounds, namely compounds 169 to 175, were prepared using the aryl ketone raw materials listed in Table 14.

[0666] Table 14: Compounds from Examples 169-175

[0667]

[0668]

[0669]

[0670] Example 176

[0671] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)-5-fluorobenzo[c]isoxazole-6-ol (compound 176)

[0672]

[0673] Step 1: Synthesis of Compound 176-1

[0674] Int-L (19 mg, 74 μmol) and 141-1 (19 mg, 82 μmol) were dissolved in DCM (1 mL), followed by the addition of TEA (23 mg, 224 μmol, 32 μL) and DMAP (7 mg, 56 μmol). The reaction was carried out at 30 °C for 6 hours. After the reaction was monitored by TLC until complete, the reaction was quenched with water, and the mixture was extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 176-1 (16 mg, 53% yield).

[0675] Step 2: Synthesis of Compound 176

[0676] 176-1 (16 mg, 40 μmol) was dissolved in DCM (1 mL), and BBr3 (100 μL, 2.0 M in DCM, 200 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 176 (1 mg, yield 6%).

[0677] 1 H NMR (400MHz, CDCl3) δ7.44(d,J=8.2Hz,2H),7.35(s,1H),7.09(d,J=8.2Hz,2H),6. 96(d,J=9.2Hz,1H),6.87(d,J=7.4Hz,1H),4.79(s,2H),3.24(s,3H).ESI-MS[M+H] + :390.1.

[0678] Example 177

[0679] Preparation of 3-(((5-(4-chlorophenyl)-4-methyl-4H-1,2,4-triazol-3-yl)thio)methyl)benzo[c]isoxazole-5,6-diol (compound 177)

[0680]

[0681] Step 1: Synthesis of Compound 177-1

[0682] Int-B (46 mg, 0.2 mmol), Int-G (55 mg, 0.2 mmol), and potassium carbonate (83 mg, 0.6 mmol) were dissolved in DMF (1 mL). The reaction was carried out at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the solution was removed under reduced pressure and purified by column chromatography to obtain 177-1 (36 mg, yield 44%).

[0683] Step 2: Synthesis of Compound 177

[0684] 177-1 (8 mg, 19 μmol) was dissolved in DCM (1 mL), and BBr3 (95 μL, 2.0 M in DCM, 190 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 177 (3 mg, yield 47%).

[0685] 1 H NMR (400MHz, Methanol-d4) δ7.57(d,J=8.3Hz,2H),7.52(d,J=8.2Hz,2H),6.57(s,1H),6.23(s,1H),4.69(s,2H),3.18(d,J=1.3Hz,3H).ESI-MS[M+H] + :389.0.

[0686] Example 178

[0687] Preparation of 3-((thieno[2,3-d]pyrimidin-2-ylthio)methyl)benzo[c]isoxazole-5,6-diol (compound 178)

[0688]

[0689] Step 1: Synthesis of Compound 178-1

[0690] 2-Chlorothio[2,3-d]pyrimidine (150 mg, 879 μmol) and thiourea (134 mg, 1.76 mmol) were dissolved in EtOH (1.5 mL). After reacting at 80 °C for 24 hours, the mixture was cooled to room temperature, and sodium hydroxide solution (5 mL, 2.0 M in H2O) was added. After reflux for 2 hours, the mixture was desoluble under reduced pressure until no alcohol odor was detected. The remaining substance was extracted twice with DCM, and the aqueous phase was collected. The pH of the aqueous phase was adjusted to 2–3 with dilute hydrochloric acid, and a solid precipitated. The solid was filtered, the filter cake was washed with water, collected, and dried to obtain crude product 178-1 (132 mg, yield 89%).

[0691] Step 2: Synthesis of Compound 178-2

[0692] 178-1 (51 mg, 0.3 mmol), Int-G (82 mg, 0.3 mmol), and potassium carbonate (125 mg, 0.9 mmol) were dissolved in DMF (1 mL). The reaction was carried out at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure and purified by column chromatography to obtain 178-2 (57 mg, yield 53%).

[0693] Step 3: Synthesis of Compound 178

[0694] 178-2 (15 mg, 41 μmol) was dissolved in DCM (1 mL), and BBr3 (205 μL, 2.0 M in DCM, 410 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the product was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 178 (3 mg, yield 20%).

[0695] 1 H NMR(600MHz,DMSO-d6)δ10.34(s,1H),9.72(s,1H),9.18(s,1H),7.69-7.95(m,1H),7.51(s,1H),6.88(s,1H),6.54(s,1H),4.93(s,2H).ESI-MS[M+H] + :332.0.

[0696] Example 179

[0697] Preparation of 3-((thieno[3,2-d]pyrimidin-2-ylthio)methyl)benzo[c]isoxazole-5,6-diol (compound 179)

[0698]

[0699] Compound 179 was prepared from 2-chlorothieno[3,2-d]pyrimidine using the synthetic method of Example 178, with an overall yield of 5% in the three-step reaction.

[0700] 1 H NMR (600MHz, DMSO-d6) δ10.28(s,1H),9.79(s,1H),9.39(s,1H),8.53(d,J=5.4Hz ,1H),7.61(d,J=5.4Hz,1H),6.93(s,1H),6.53(s,1H),4.91(s,2H).ESI-MS[M+H] + :332.0.

[0701] Example 180

[0702] Preparation of 3-((1H-indazol-1-yl)methyl)benzo[c]isoxazole-5,6-diol (compound 180)

[0703]

[0704] Step 1: Synthesis of Compound 180-1

[0705] Indazole (36 mg, 0.3 mmol), Int-G (82 mg, 0.3 mmol), and cesium carbonate (293 mg, 0.9 mmol) were dissolved in DMF (1 mL). The reaction was carried out at room temperature for 16 hours. After the reaction was completed by TLC monitoring, the solution was removed under reduced pressure and purified by column chromatography to obtain 180-1 (36 mg, yield 38%).

[0706] Step 2: Synthesis of Compound 180

[0707] 180-1 (36 mg, 116 μmol) was dissolved in DCM (1 mL), and BBr3 (580 μL, 2.0 M in DCM, 1.16 mmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the product was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 180 (3 mg, yield 9%).

[0708] 1 H NMR (600MHz, DMSO-d6) δ10.38(s,1H),9.75(s,1H),8.14(s,1H),7.78(dd,J=15.6,8.3Hz,2H ),7.40-7.49(m,1H),7.12-7.24(m,1H),6.55(s,1H),6.46(s,1H),6.12(s,2H).ESI-MS[M+H] + :282.1.

[0709] Example 181

[0710] Preparation of 3-((4-(4-chlorophenyl)-1H-pyrazol-1-yl)methyl)benzo[c]isoxazole-5,6-diol (compound 181)

[0711]

[0712] Compound 181 was prepared from 4-(4-chlorophenyl)-1H-pyrazole using the synthetic method of Example 180, with an overall yield of 4% in the two-step reaction.

[0713] 1H NMR(600MHz,DMSO-d6)δ12.74(s,1H),9.71(s,1H),9.40(s,1H),8.09(s,1H),7.75(d,J=8.1Hz,1H), 7.66(d,J=8.4Hz,1H),7.30-7.47(m,1H),7.06-7.19(m,1H),6.80(s,1H),5.83(s,2H).ESI-MS[M+H] + :342.1.

[0714] Example 182

[0715] Preparation of 3-(((3-(4-chlorophenyl)isoxazole-5-yl)oxy)methyl)benzo[c]isoxazole-5,6-diol (compound 182)

[0716]

[0717] Step 1: Synthesis of Compound 182-1

[0718] 3-(4-chlorophenyl)-5-hydroxyisoxazole (30 mg, 153 μmol) was dissolved in DMF (0.5 mL). After cooling to 0 °C, sodium hydride (20 mg, 500 μmol, 60% mineral oil) was added. After stirring for 30 minutes, Int-G (42 mg, 153 μmol) was added. The reaction was carried out at 30 °C for 2 hours. After the reaction was completed by TLC monitoring, methanol was added to quench the reaction, the solvent was removed under reduced pressure, and the product was purified by column chromatography to obtain 182-1 (20 mg, 33% yield).

[0719] Step 2: Synthesis of Compound 182

[0720] 182-1 (20 mg, 51 μmol) was dissolved in DCM (1 mL), and BBr3 (255 μL, 2.0 M in DCM, 510 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the product was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 182 (2 mg, yield 10%).

[0721] 1 H NMR(400MHz,Methanol-d4)δ7.63-7.50(m,5H),6.59(s,1H),6.52(s,1H),4.10(s,2H).ESI-MS[M+H] + :359.1.

[0722] Example 183

[0723] Preparation of 3-((((4-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)methyl)amino)methyl)benzo[c]isoxazole-5,6-diol (compound 183)

[0724]

[0725] Step 1: Synthesis of Compound 183-1

[0726] Boc-glycine (3 g, 17.1 mmol) and cesium carbonate (3.25 g, 10 mmol) were dispersed in DMF (60 mL). The reaction was carried out at room temperature until the cesium carbonate was completely dissolved, and the generated ethanol was removed under reduced pressure. 2-Bromo-4'-chloroacetophenone (4 g, 17.1 mmol) and DMF (60 mL) were added under nitrogen protection. The reaction was carried out at room temperature for 2 hours, and the solvent was removed under reduced pressure. The solution was extracted with EA and water. The organic phase was dried and purified by column chromatography to give 183-1 (5.1 g, 89% yield).

[0727] Step 2: Synthesis of Compound 183-2

[0728] 183-1 (5.1 g, 15.5 mmol) and ammonium acetate (4.8 g, 62 mmol) were dissolved in xylene. The reaction flask was fitted with a water separator and refluxed at 180 °C until no water was separated. After cooling to room temperature, EA (100 mL), sodium bicarbonate solution (100 mL, 5% inH2O), and water (100 mL) were added sequentially. After drying the organic phase, the solvent was removed under reduced pressure. After slurrying with petroleum ether, 183-2 (2.7 g, yield 56%) was obtained.

[0729] Step 3: Synthesis of Compound 183-3

[0730] 183-2 (2.7 g, 8.77 mmol) and potassium carbonate (1.25 g, 9 mmol) were dissolved in DMF (30 mL). After cooling to 0 °C, iodomethane (1.78 g, 12.5 mmol, 785 μL) was added. The reaction was carried out at 60 °C for 24 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water and extracted three times with DCM. After drying the organic phase, the solvent was removed under reduced pressure and purified by column chromatography to obtain 183-3 (882 mg, yield 31%).

[0731] Step 4: Synthesis of compound 183-4

[0732] 183-3 (882 mg, 2.74 mmol) was dissolved in TFA (1.5 mL). The reaction was allowed to proceed at room temperature for 30 minutes, and then quenched with water. The pH was adjusted to 7-8 with saturated potassium carbonate solution, resulting in the precipitation of a large amount of solid. The mixture was filtered, and the filter cake was pulped with methyl tert-butyl ether to give 183-4 (560 mg, 86% yield).

[0733] Step 5: Synthesis of compound 183-5

[0734] Int-G (62 mg, 229 μmol), 183-4 (59 mg, 263 μmol), and DIPEA (150 mg, 1.15 mmol, 200 μL) were dissolved in DMF (1 mL). The reaction was carried out overnight at room temperature. After the reaction was monitored by TLC until complete, the reaction was quenched by adding saturated ammonium chloride solution and extracted three times with EA. The organic phase was dried and dissolved under reduced pressure, and purified by column chromatography to obtain 183-5 (42 mg, yield 44%).

[0735] Step 6: Synthesis of Compound 183

[0736] 183-5 (30 mg, 72 μmol) was dissolved in DCM (1 mL), and BBr3 (360 μL, 2.0 M in DCM, 720 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 183 (15 mg, yield 50%).

[0737] 1 H NMR (400MHz, Methanol-d4) δ7.57(d,J=8.6Hz,2H),7.52(s,1H),7.43(d,J=8.6Hz, 2H),6.69(s,1H),6.47(s,1H),4.52(s,2H),4.29(s,2H),3.80(s,3H).ESI-MS[M+H] + :385.1.

[0738] Example 184

[0739] Preparation of 3-(((5-(4-chlorophenyl)-1-methyl-1H-imidazol-2-yl)thio)methyl)-1H-indazole-5,6-diol (compound 184)

[0740]

[0741] Step 1: Synthesis of Compound 184-1

[0742] Int-H (40 mg, 146 μmol), 141-1 (40 mg, 175 μmol), and potassium carbonate (61 mg, 438 μmol) were dissolved in DMF (0.5 mL). The reaction was carried out at 30 °C for 3 hours. After the reaction was completed by TLC monitoring, the reaction was quenched with water and extracted three times with DCM. After drying and solvent removal of the organic phase, 184-1 (34 mg, yield 57%) was purified by column chromatography.

[0743] Step 2: Synthesis of Compound 184-2

[0744] 184-1 (30 mg, 71 μmol) was dissolved in concentrated hydrochloric acid (1 mL). After cooling to -10 °C, sodium nitrite (10 mg, 142 μmol) was added. After reacting at -10 °C for 1 hour, stannous chloride dihydrate (81 mg, 359 μmol) was added. The reaction was continued at -10 °C for another hour. After the reaction was monitored by TLC until complete, the reaction was quenched with ice water. The precipitated solid was filtered, the filter cake was washed with water, and the collected filter cake was purified by column chromatography to obtain 184-2 (10 mg, 31% yield).

[0745] Step 3: Synthesis of Compound 184

[0746] 184-2 (10 mg, 23 μmol) was dissolved in DCM (1 mL), and BBr3 (115 μL, 2.0 M in DCM, 230 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 184 (4 mg, yield 42%).

[0747] 1 H NMR(600MHz,DMSO-d6)δ12.35(s,1H),9.42(s,1H),8.82(s,1H),7.59(d,J=8.3Hz,3H), 7.47(d,J=8.4Hz,2H),6.80(s,1H),6.75(s,1H),4.55(s,2H),3.34(s,3H).ESI-MS[M+H] + :387.1.

[0748] Examples 185-187

[0749] Referring to the synthesis method of Example 184, the corresponding compounds, namely compounds 185 to 187, were prepared using the mercaptoimidazole raw material and α-bromoacetophenone raw material listed in Table 15.

[0750] Table 15: Compounds from Examples 185-187

[0751]

[0752] Example 188

[0753] Preparation of 3-((1H-indazol-1-yl)methyl)-7-bromo-1H-indazol-5,6-diol (compound 188)

[0754]

[0755] Step 1: Synthesis of Compound 188-1

[0756] Indazole (16 mg, 136 μmol) was dissolved in DMF (0.8 mL), and cesium carbonate (37 mg, 113 μmol) was added. After reacting at room temperature for 30 minutes, Int-I (40 mg in 0.8 mL DMF, 113 μmol) was added. The reaction was continued at 30 °C for 15 hours. After the reaction was monitored by TLC until complete, the reaction was quenched with saturated brine, and the mixture was extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 188-1 (18 mg, 40% yield).

[0757] Step 2: Synthesis of Compound 188-2

[0758] Dissolve 188-1 (18 mg, 46 μmol) in concentrated hydrochloric acid (0.5 mL). Cool to -10 °C and add sodium nitrite (7 mg, 92 μmol). After reacting at -10 °C for 1 hour, add stannous chloride dihydrate (52 mg, 231 μmol). Continue reacting at -10 °C for 1 hour. After the reaction is complete as monitored by TLC, quench the reaction with ice water. The precipitated solid is filtered, the filter cake is washed with water, collected, and purified by column chromatography to obtain 188-2 (8 mg, yield 44%).

[0759] Step 3: Synthesis of Compound 188

[0760] 188-2 (8 mg, 20 μmol) was dissolved in DCM (0.5 mL), and BBr3 (100 μL, 2.0 M in DCM, 200 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 188 (4 mg, yield 55%).

[0761] 1 H NMR (600MHz, DMSO-d6) δ8.39(s,1H),7.99(s,1H),7.63(d,J=8.5Hz,2H),7.42(d,J=8.5Hz,2H),6.57(s,1H),6.51(s,1H),5.81(s,2H).ESI-MS[M+H] + :359.0.

[0762] Examples 189-200

[0763] Referring to the synthesis method of Example 188, the corresponding compounds, namely compounds 189 to 200, were prepared using the amine raw materials listed in Table 16.

[0764] Table 16: Compounds from Examples 188-200

[0765]

[0766]

[0767]

[0768]

[0769] Example 201

[0770] Preparation of 3-((1H-indazol-1-yl)methyl)-4,7-dibromo-1H-indazol-5,6-diol (compound 201)

[0771]

[0772] Step 1: Synthesis of Compound 201-1

[0773] 188-2 (20 mg, 51 μmol) was dissolved in ultradry THF (1 mL), and then sodium hydride (13 mg, 510 μmol, 60% mineral oil) was added. After 10 minutes, acryloyl chloride (23 mg, 254 μmol, 21 μL) was added. The reaction was allowed to proceed at room temperature for 2 hours. After the reaction was confirmed to be complete by TLC, the reaction was carefully quenched with water, extracted with DCM, dissolved under reduced pressure, and purified by column chromatography to obtain 201-1 (8 mg, 37% yield).

[0774] Step 2: Synthesis of Compound 201

[0775] 201-1 (8 mg, 18 μmol) was dissolved in DCM (0.5 mL), and BBr3 (90 μL, 2.0 M in DCM, 180 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 201 (3 mg, yield 37%).

[0776] 1 H NMR(600MHz,DMSO-d6)δ13.07(s,1H),10.00(s,1H),9.38(s,1H),8.01(s,1H),7.74(d,J=8.1Hz,1 H),7.64(d,J=8.5Hz,1H),7.34(t,J=7.6Hz,1H),7.12(t,J=7.4Hz,1H),6.05(s,2H).ESI-MS[M+H] + :438.9.

[0777] Example 202

[0778] Preparation of 2-((4,7-dibromo-5,6-dihydroxy-1H-indazol-3-yl)methyl)isoindoline-1-one (compound 202)

[0779]

[0780] Step 1: Synthesis of Compound 202-1

[0781] Isoindoline-1-one (80 mg, 601 μmol) was dissolved in ultradry THF (3 mL), and then sodium hydride (15 mg, 601 μmol, in 60% mineral oil) was added. After 10 minutes, Int-I (213 mg, 601 μmol) was added. The reaction was allowed to proceed at room temperature for 2 hours. After the reaction was confirmed to be complete by TLC, the reaction was carefully quenched with water, extracted with DCM, dissolved under reduced pressure, and purified by column chromatography to obtain 202-1 (70 mg, 28% yield).

[0782] Step 2: Synthesis of compound 202-2

[0783] Dissolve 202-1 (70 mg, 172 μmol) in concentrated hydrochloric acid (2 mL). Cool to -10 °C and add sodium nitrite (36 mg, 518 μmol). After reacting at -10 °C for 1 hour, add stannous chloride dihydrate (164 mg, 863 μmol). Continue reacting at -10 °C for another hour. After the reaction is complete as monitored by TLC, quench the reaction with ice water. The precipitated solid is filtered, the filter cake is washed with water, collected, and purified by column chromatography to obtain 202-2 (19 mg, 28% yield).

[0784] Step 3: Synthesis of Compound 202

[0785] Compound 202-2 (7 mg, 17 μmol) was dissolved in DCM (0.5 mL), and BBr3 (85 μL, 2.0 M in DCM, 170 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the compound was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 202 (1.5 mg, yield 17%).

[0786] 1 H NMR (600MHz, DMSO-d6) δ13.01(s,1H),10.01(s,1H),9.40(s,1H),7.72(d,J=7.7Hz,1H),7.57( dt,J=13.8,4.8Hz,2H),7.49(t,J=7.2Hz,1H),5.20(d,J=2.3Hz,2H),4.42(s,2H).ESI-MS[M+H] + :454.9.

[0787] Example 203

[0788] Preparation of 3-((1H-indazol-1-yl)methyl)-7-(thiophen-2-yl)-1H-indazol-5,6-diol (compound 203)

[0789]

[0790] Step 1: Synthesis of Compound 203-1

[0791] 188-2 (15 mg, 38 μmol), thiophene-2-boric acid (25 mg, 193 μmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (7 mg, 8 μmol), and potassium carbonate (16 mg, 116 μmol) were dissolved in a mixed solvent of 1,4-dioxane (0.8 mL) and water (0.2 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 110 °C. After the reaction was monitored by TLC until complete, the insoluble matter was filtered off, the reaction was quenched with water, and extracted with EA. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 203-1 (12 mg, 79% yield).

[0792] Step 2: Synthesis of Compound 203

[0793] 203-1 (10 mg, 25 μmol) was dissolved in DCM (0.5 mL), and BBr3 (125 μL, 2.0 M in DCM, 250 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the product was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 203 (3 mg, yield 32%).

[0794] 1 H NMR (600MHz, DMSO-d6) δ12.49(s,1H),9.67(s,1H),9.15(s,1H),8.10(s,1H),7.76(d,J=8.1Hz,1H),7.70(d,J=8.5Hz,1H),7.59(d,J=5.1Hz, 1H),7.50(d,J=3.6Hz,1H),7.36(t,J=7.7Hz,1H),7.17(dd,J=5.1,3.7Hz,1H),7.13(t,J=7.4Hz,1H),6.87(s,1H),5.86(s,2H).ESI-MS[M+H] + :363.1.

[0795] Examples 204-209

[0796] Referring to the synthesis method of Example 203, the corresponding compounds, namely compounds 204 to 209, were prepared using the boric acid (ester) raw materials listed in Table 17.

[0797] Table 17: Compounds from Examples 204-209

[0798]

[0799]

[0800] Example 210

[0801] Preparation of 3-((1H-indazol-1-yl)methyl)-7-morpholino-1H-indazol-5,6-diol (compound 210)

[0802]

[0803] Step 1: Synthesis of Compound 210-1

[0804] 188-2 (15 mg, 38 μmol), morpholine (34 mg, 387 μmol, 34 μL), methanesulfonic acid (2-di-tert-butylphosphine-2',4',6'-triisopropylbiphenyl)(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (7 mg, 8 μmol), 2-di-tert-butylphosphine-2',4',6'-triisopropylbiphenyl (4 mg, 8 μmol), and sodium tert-butoxide (12 mg, 117 μmol) were dissolved in 1,4-dioxane (1 mL). After purging the gas with nitrogen six times, the reaction was carried out overnight at 110 °C. After the reaction was monitored by TLC until complete, the insoluble matter was filtered off, and the mixture was extracted with water and EA. The organic phase was dried, filtered, evaporated to dryness, and purified by column chromatography to obtain 210-1 (3 mg, 19% yield).

[0805] Step 2: Synthesis of Compound 210

[0806] 210-1 (3 mg, 7 μmol) was dissolved in DCM (0.5 mL), and BBr3 (35 μL, 2.0 M in DCM, 70 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 210 (1 mg, yield 28%).

[0807] 1H NMR (600MHz, Methanol-d4) δ8.03(s,1H),7.73(d,J=8.1Hz,1H),7.59(d,J=8.5Hz,1H),7.35(t,J=7.7 Hz,1H),7.12(t,J=7.5Hz,1H),6.63(s,1H),5.84(s,2H),3.84-3.99(m,4H),3.25(s,4H).ESI-MS[M+H] + :366.1.

[0808] Example 211

[0809] Preparation of 3-((1H-indazol-1-yl)methyl)-7-(piperazin-1-yl)-1H-indazol-5,6-diol (compound 211)

[0810]

[0811] Compound 211 was prepared from 1-Boc-piperazine using the synthetic method of Example 210, with an overall yield of 22% in the two-step reaction.

[0812] 1 H NMR (400MHz, DMSO-d6) δ12.42(s,1H),9.44(s,1H),8.56(s,3H),8.08(s,1H),7.75(d,J=8.2Hz,1H),7.67(d,J=8.4Hz ,1H),7.35(t,J=7.6Hz,1H),7.11(t,J=7.5Hz,1H),6.66(s,1H),5.81(s,2H),3.25(s,4H),2.55(s,4H).ESI-MS[M+H] + 365.2.

[0813] Example 212

[0814] Preparation of 3-((1H-indazol-1-yl)methyl)-7-bromo-1-methyl-1H-indazol-5,6-diol (compound 212)

[0815]

[0816] Step 1: Synthesis of compound 212-1

[0817] 188-2 (15 mg, 38 μmol), methyl iodoform (28 mg, 193 μmol, 13 μL), and cesium carbonate (16 mg, 46 μmol) were dissolved in DMF (0.5 mL). The reaction was carried out at 50 °C for 4 hours. After the reaction was completed by TLC monitoring, the solvent was removed under reduced pressure, and the solution was purified by column chromatography to obtain 212-1 (10 mg, yield 64%).

[0818] Step 2: Synthesis of Compound 212

[0819] 212-1 (10 mg, 24 μmol) was dissolved in DCM (0.5 mL), and BBr3 (120 μL, 2.0 M in DCM, 240 μmol) was added. The reaction was carried out at 30 °C for 1 hour. After the reaction was completed by LC-MS, the solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 212 (5 mg, yield 53%).

[0820] 1 H NMR (600MHz, DMSO-d6) δ9.87 (s, 1H), 9.48 (s, 1H), 8.09 (d, J = 0.8Hz, 1H), 7.75 (d, J = 8.1Hz, 1H), 7.60-7.73 (m, 1H),7.38(ddd,J=8.2,6.9,0.9Hz,1H),7.08-7.18(m,1H),6.84(s,1H),5.80(s,2H),4.20(s,3H).ESI-MS[M+H] + :373.0.

[0821] Examples 213-220

[0822] Referring to the synthesis method of Example 212, the corresponding compounds, namely compounds 204 to 220, were prepared using the halogenated raw materials in Table 18.

[0823] Table 18: Compounds from Examples 213-220

[0824]

[0825]

[0826]

[0827]

[0828] Example 221

[0829] Preparation of 7-bromo-1-(2-hydroxyethyl)-3-(indoline-1-ylmethyl)-1H-indazole-5,6-diol (compound 221)

[0830]

[0831] Step 1: Synthesis of compound 221-1

[0832] Indoline (50 mg, 420 μmol) was dissolved in DMF (1 mL), and cesium carbonate (137 mg, 420 μmol) was added. After reacting at room temperature for 30 minutes, Int-I (149 mg in 1 mL DMF, 420 μmol) was added. The reaction was continued at 30 °C for 15 hours. After the reaction was complete as monitored by TLC, the reaction was quenched with saturated brine, and the mixture was extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 221-1 (80 mg, yield 48%).

[0833] Step 2: Synthesis of compound 221-2

[0834] 221-1 (80 mg, 203 μmol) was dissolved in concentrated hydrochloric acid (1 mL). After cooling to -10 °C, sodium nitrite (43 mg, 610 μmol) was added. After reacting at -10 °C for 1 hour, stannous chloride dihydrate (193 mg, 1.02 mmol) was added. The reaction was continued at -10 °C for another 1 hour. After the reaction was monitored by TLC until complete, the reaction was quenched with ice water. The precipitated solid was filtered, the filter cake was washed with water, and the filter cake was collected and purified by column chromatography to obtain 221-2 (50 mg, 40% yield).

[0835] Step 3: Synthesis of compound 221-3

[0836] 221-2 (20 mg, 51 μmol) was dissolved in DMF (0.5 mL), and cesium carbonate (17 mg, 51 μmol) was added. After reacting at room temperature for 30 minutes, 2-bromoethyl methyl ether (8 mg in 0.5 mL DMF, 51 μmol) was added. The reaction was continued at 30 °C for 15 hours. After the reaction was complete as monitored by TLC, the reaction was quenched with saturated brine, and the mixture was extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 221-3 (10 mg, 43% yield).

[0837] Step 4: Synthesis of Compound 221

[0838] 221-3 (5 mg, 11 μmol) was dissolved in DCM (0.5 mL), and BBr3 (55 μL, 2.0 M in DCM, 110 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 221 (1.2 mg, yield 26%).

[0839] 1H NMR(600MHz,DMSO-d6)δ9.79(s,1H),9.46(s,1H),7.05(s,1H),7.04-6.98(m,2H),6.74(d,J=7.9Hz,1H),6.59(t,J=7.3Hz,1H),4 .66(t,J=6.7Hz,2H),4.40(s,2H),3.71(t,J=6.7Hz,2H),3.22(t,J=8.3Hz,2H),3.17(s,1H),2.84(t,J=8.3Hz,2H).ESI-MS[M+H] + :404.1.

[0840] Examples 222-228

[0841] Referring to the synthesis method of Example 221, the corresponding compounds, namely compounds 222 to 228, were prepared using the amine raw materials listed in Table 19.

[0842] Table 19: Compounds from Examples 222-228

[0843]

[0844]

[0845]

[0846] Example 229

[0847] Preparation of 2-((4,7-dibromo-5,6-dihydroxy-1-(2-hydroxyethyl)-1H-indazol-3-yl)methyl)isoindoline-1-one (compound 229)

[0848]

[0849] Step 1: Synthesis of Compound 229-1

[0850] 202-2 (11 mg, 25 μmol) was dissolved in DMF (0.5 mL), and cesium carbonate (9 mg, 25 μmol) was added. After reacting at room temperature for 30 minutes, 2-bromoethyl methyl ether (14 mg in 0.5 mL DMF, 100 μmol) was added. The reaction was continued at 30 °C for 15 hours. After the reaction was complete as monitored by TLC, the reaction was quenched with saturated brine, and the mixture was extracted three times with DCM. The organic phase was dried, dissolved under reduced pressure, and purified by column chromatography to obtain 229-1 (8 mg, 69% yield).

[0851] Step 2: Synthesis of Compound 229

[0852] 229-1 (6 mg, 13 μmol) was dissolved in DCM (0.5 mL), and BBr3 (65 μL, 2.0 M in DCM, 130 μmol) was added. The reaction was carried out at 30 °C for 1 hour. The reaction was monitored by LC-MS until completion. The solvent was removed under reduced pressure at room temperature, and the solution was purified by reverse semi-preparative high performance liquid chromatography to obtain compound 229 (1.6 mg, yield 24%).

[0853] 1 H NMR (600MHz, DMSO-d6) δ10.02(s,1H),9.52(s,1H),7.72(d,J=7.0Hz,1H),7.57(d,J=6.7Hz,2H),7.49(t, J=7.3Hz,1H),5.21(s,2H),4.81(s,1H),4.74-4.62(m,2H),4.42(s,2H),3.73-3.65(m,2H).ESI-MS[M+H] + :497.7.

[0854] Test Example 1: Compound Enzyme Activity Test

[0855] DNA unwinding assay: The test compound was first diluted in reaction buffer (50 mM Tris-HCl pH 8.0, 5 mM NaCl, 2 mM MgCl2, 0.01% Tween-20, 1 mM DTT, 2.5 μg / ml poly dIC) in a 3-fold serial dilution, resulting in 11 concentrations. Then, 5 μL of the diluted compound was mixed with 10 μL of protein and incubated at room temperature for 15 minutes. Next, 2.5 μL of forked DNA and 2.5 μL of ATP-Mg substrate were added to initiate the reaction, and BioTek was used. TM Synergy TM The Neo2 microplate reader was used to detect and record fluorescence signals. The reaction was carried out at 25°C in a 384-well plate with a total reaction volume of 20 μL. The maximum compound concentration was 100 μM, the final concentration of protein was 30 nM, the final concentration of DNA was 90 nM, the final concentration of ATP-Mg was 1 mM, and the final concentration of DMSO was 1%.

[0856] The sequence of the forked DNA is:

[0857] 5'-TAMRA-GAACGAACACATCGGGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-3',5'-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGATGTGTTCGTTC-BHQ2-3'. The fluorescence signal value after 20 minutes of reaction was taken, and the background signal value was subtracted before calculation.

[0858] The activity data were calculated as follows: Activity% = (Fluorescence Sample / Fluorescence DMSO) × 100%, and the inhibition rate was calculated as follows: Inhibition% = 100% - Activity%. IC50 was fitted using the dose-response-inhibition (variable slope) method in Graphpad Prism software.

[0859] The test results are listed in Table 20.

[0860] Table 20: Inhibitory activity of the compounds of the present invention against WRN helicase (IC50, 100 mg / L) 50 )

[0861]

[0862]

[0863]

[0864]

[0865] Where A ≤ 1 μM; 1 μM <B≤10μΜ;10μΜ<C≤30μΜ;D> 30μM.

[0866] As can be seen from the above, the compounds of the present invention have a strong WRN inhibitory effect and can be used as drugs for treating MSI tumors or cancer.

[0867] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A compound of Formula I, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Formula I R1is selected from the group consisting of: R2, R3, R4, R5, and R6are each independently selected from the group consisting of: L is an optionally substituted linking group selected from a saturated or partially unsaturated straight chain C 1-13 heteroalkylene; R1is selected from -C(O)-, -C(O)-CH2-, pyridinylidene, -NH-, -CH2-, and -H, -OH, halogen (e.g., -F, -CI, -Br, -I), -N02, -COOH, and -B(OH)2; an optionally substituted 5-6 membered saturated or partially unsaturated heterocyclyl or heteroaryl, wherein the optionally substituted 5-6 membered saturated or partially unsaturated heterocyclyl or heteroaryl comprises one or more heteroatoms selected from O, N, or S, and when comprising multiple heteroatoms, the heteroatoms are the same or different; or optionally substituted C 1-5 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 cycloalkenyl, optionally substituted C 3-5 alkenyl, C 1-5 alkoxy, and -O-C 1-5 alkanoyl; R1and R2, or R1and R6, together with the carbon atoms to which they are each attached, form a saturated or partially unsaturated 5-6 membered heterocyclic ring comprising one or two heteroatoms selected from O, N, and S. -NR a R b -C(O)NR a R b , where R a and R b Each is independently selected from -H, -OH, and -C. 1-5 Alkyl group; Optionally, R4is connected with one of R3or R5to form an optionally substituted saturated or unsaturated straight chain C 1-5 heteroalkylene comprising 1 or 2 heteroatoms selected from O or N; 1-5 heteroalkylene comprising 1 or 2 heteroatoms selected from O or N; 2. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein:

3. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: L is a linking group represented by the formula "-D x -(CH2) p -(C(O)) y -(NR L ) z -(CH2) w -” wherein D is an atom selected from O and S, and x, y and z are each independently selected from 0 or 1, w and p are each independently integers selected from 0-10 and w+p < 10, wherein R L is selected from -H, optionally substituted C 1-5 alkyl, preferably, w and p are each independently selected from 0, 1, 2, 3, 4 or 5. R2and R6are each independently selected from the group consisting of: -H, -OH, halogen (e.g., -F, -CI, -Br, -I), or -NH2, provided that R2and R6are both -H, or R2and R6are not the same.

4. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R3and R5are each independently selected from the group consisting of: -H, -OH, halogen (e.g., -F, -CI, -Br, -I), -N02, and -COOH; an optionally substituted 5-6 membered saturated or partially unsaturated heterocyclyl or heteroaryl, wherein the heterocyclyl or heteroaryl comprises one or more heteroatoms selected from O, N, or S, and when comprising multiple heteroatoms, the heteroatoms are the same or different; or optionally substituted C 1-5 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 cycloalkenyl, optionally substituted C 3-5 alkenyl, C 1-5 alkoxy, and -O-C 1-5 alkanoyl; 5. The compound of claim 4, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: -NR a R b 、-C(O)NR a R b , -C(O)OR a and R b are each independently selected from the group consisting of -H, -OH, and -C 1-5 alkanoyl. R3and R5are each independently selected from the group consisting of: ​ optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted cycloheptyl, optionally substituted cyclooctyl, optionally substituted cyclononyl, optionally substituted cyclodecyl, optionally substituted cyclobutenyl, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cycloheptenyl, optionally substituted cyclooctenyl, optionally substituted cyclononetenyl, optionally substituted cyclodecetenyl, optionally substituted propenyl, optionally substituted butenyl, optionally substituted pentenyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-valeryl; or optionally substituted thienyl, optionally substituted thiazolyl, optionally substituted pyridyl, optionally substituted morpholinyl, optionally substituted piperazinyl; or -NR a R b , -C(O)NR a R b , -C(O)OR a , -C(O)H, -CN, -NO2, -F, -Cl, -Br, and -I. In some embodiments, R b each independently is selected from the group consisting of -H, -OH, and formyl, acetyl, propionyl, butyryl, or valeryl.

6. The compound of claim 5, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R3and R5are each independently selected from the group consisting of: -CH3, -OCH3, -OC(0)CH3, -OC(0)CH2CH3, -C3H5(e.g., isopropenyl), cyclopropyl, cyclohexenyl substituted with -OH (e.g., ), -NH2, -CONH2, -CONHOH, 7. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R4is selected from the group consisting of: -H, -OH, -B(OH)2, halogen (e.g., -F, -Cl, -Br, -I); optionally substituted methyl, optionally substituted ethyl, optionally substituted propyl, optionally substituted butyl, optionally substituted pentyl, methoxy, ethoxy, propoxy, butoxy, pentoxy, -O-formyl, -O-acetyl, -O-propionyl, -O-butyryl, or -O-valeryl; or -NR a R b , -C(O)NR a R b , -C(O)NR a and R b are each independently selected from the group consisting of -H, -OH, and -formyl, acetyl, propionyl, -butyryl, or valeryl.

8. The compound of claim 7, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R4 is selected from: -CH3, -OCH3, -OCOCH3, -OCOCH2CH3, -NH2, or -NHCOCH3.

9. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R4and R3are linked to form a heteroalkylene selected from -Q- optionally substituted C 1-5 alkylene-Q-, -Q- optionally substituted C 1-5 alkylene-Q-, -Q- optionally substituted C 1-5 alkylene-Q-, -Q- optionally substituted C 1-5 alkylene-Q-, -Q- optionally substituted C 10. The compound of claim 9, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: the group formed by the connection of R4and R3is selected from:

11. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Ring A represents optionally substituted: wherein 1-3 of G1-G5 are each independently selected from N or O, with the remainder being C; 1-2 of I1-I4 are N, with the remainder being C; 1-3 of J1-J6 are N, with the remainder being C; 1-2 of K1-K3 are N, with the remainder being C, and 0-2 of K4-K7 are N, with the remainder being C; 1-2 of L1-L3 are N, with the remainder being C; 1 of M1-M2 is S, with the remainder being C; represents a single bond, a double bond, or the ring is aromatic between adjacent atoms of the ring.

12. The compound of claim 11, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Ring A represents: wherein 1 to 3 of G1, G3and G4are N, and the others are CH; or Ring A represents: wherein one of G1, G3, G4and G5is O, one is N, and the rest are C; or Ring A represents: wherein one of G1, G3, G4and G5is N, and the others are C; or Ring A represents wherein 1 to 3 of J2-J6are N, the others are C; or Ring A represents: wherein 1 to 2 of J2-J6are N, the others are C; or Ring A represents wherein one of K1-K2is N and the others are C, and 0 to 2 of K4-K7are N and the others are C; or Ring A represents: wherein 1 to 2 of K1and K3are N, and the remainder are C, and 0 to 2 of K4-K7are N, and the remainder are C.

13. The compound of claim 12, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Ring A is optionally substituted with R7 and / or R8, wherein, R7 is selected from the following groups: -H, =0, halogen (e.g., -F, -CI, -Br, -I); optionally substituted C 1-5 alkyl, optionally substituted C 3-10 cycloalkyl, optionally substituted C 1-5 alkenyl, optionally substituted C 1-5 alkynyl, optionally substituted C 1-5 alkoxy, optionally substituted C 1-5 alkylene-C 1-5 alkoxy; optionally substituted phenyl, optionally substituted 5-6 membered heterocyclyl, or heteroaryl; optionally substituted C 1-5 alkylene-NR c R d , -NR c R d , wherein R c and R d are each independently selected from -H or C 1-5 alkyl; and C 1-5 alkylene-5-6 membered heterocyclyl or heteroaryl; and R8 is selected from the following groups: -H, halogen (-F, -CI, -Br, -I); optionally substituted C 1-5 alkyl, -C 1-5 alkoxy; -C(O)-NR c R d wherein R c and R d each independently is selected from -H or C 1-5 alkyl; optionally substituted phenyl, optionally substituted 5-6 membered heterocyclyl, or heteroaryl; optionally substituted benzo 5-6 membered heterocyclyl or heteroaryl, optionally substituted 5-6 membered heterocyclic or heteroaromatic ring biphenyl; or wherein Z is O or S.

14. The compound of claim 13, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R7is selected from optionally substituted C 7s substituted C 1-5 alkyl, optionally substituted with one or more R 7s substituted C 3-10 cycloalkyl, optionally substituted with one or more R 7s substituted C 1-5 alkenyl, optionally substituted with one or more R 7s substituted C 1-5 alkynyl, optionally substituted with one or more R 7s substituted C 1-5 alkoxy, optionally substituted with one or more R 7s substituted -C 1-5 alkylene-C 1-5 alkoxy, optionally substituted with one or more R 7s substituted phenyl, wherein R 7s is selected from halogen, methyl, ethyl, propyl, butyl, pentyl, -CN and phenyl; or R7is selected from optionally substituted C 7s substituted C 1-5 alkylene-NR c R d , -NR c R d wherein R c and R d are each independently selected from -H, methyl, ethyl, propyl, butyl, and pentyl, wherein R 7s is selected from halogen, methyl, ethyl, propyl, butyl, pentyl, -CN, and phenyl; or R7is selected from C 1-5 alkylene-pyridinyl, or, R7is selected from optionally substituted 7s substituted wherein 1 to 2 of Y1-Y3are a heteroatom selected from N, O, n is 1 or 2, and denotes that a single, double bond or the ring is aromatic between the adjacent atoms of the ring, wherein R 7s selected from halogen, methyl, ethyl, propyl, butyl, pentyl or cyano; preferably, selected from: preferably, R7is selected from optionally substituted C 7s substituted C 1-5 alkyl, optionally substituted with 1 or 2 R 7s substituted C 3-10 cycloalkyl, optionally substituted with 1 or 2 R 7s substituted C 1-5 alkenyl, optionally substituted with 1 or 2 R 7s substituted C 1-5 alkynyl, optionally substituted with 1 or 2 R 7s substituted C 1-5 alkoxy, optionally substituted with 1 or 2 R 7s substituted -C 1-5 alkylene-C 1-5 alkoxy, optionally substituted with 1 or 2 R 7s substituted phenyl; or R7is selected from optionally substituted C 7s substituted C 1-5 alkylene-NR c R d -NR c R d wherein R c and R d each independently is selected from -H, methyl, ethyl, propyl, butyl, and pentyl; or R7is selected from optionally substituted 7s substituted wherein 1 to 2 of Y1-Y3are a heteroatom selected from N, O, n is 1 or 2, and denotes that a single, double bond or the ring is aromatic between the adjacent atoms of the ring, preferably, selected from:

15. The compound of claim 14, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R7is selected from: -F, =0, methyl, ethyl, propyl (e.g., n-propyl, isopropyl), cyclopropyl, cyclopentyl, cyclohexyl, phenyl, benzyl, dimethylaminoethyl, methoxyethyl, methylaminoethyl, cyanomethyl, allyl, cyanophenyl, 16. The compound of claim 13, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R8is selected from the group consisting of: optionally substituted by one or more R 8S1 or R 8S2 substituted phenyl; or optionally substituted with one or more R 8S1 or R 8S2 substituted 5-6 membered heterocyclyl or heteroaryl; or optionally substituted by one or more R 8S1 or R 8S2 substituted phenyl, benzo5-6 membered heterocyclyl, or heteroaryl; or optionally substituted by one or more R 8S1 and / or R 8S2 a 5-6 membered heterocyclic or heteroaromatic ring; or optionally substituted by one or more R 8S1 and / or R 8S2 a 5-6 membered heterocyclic or heteroaromatic ring and a 5-6 membered heterocyclyl or heteroaryl group; Preferably, R8is selected from the group consisting of: optionally substituted by 1 or 2 R 8S1 or R 8S2 substituted phenyl; or optionally substituted with 1 or 2 R 8S1 or R 8S2 substituted 5-6 membered heterocyclyl or heteroaryl; or optionally substituted by 1 or 2 R 8S1 or R 8S2 substituted phenyl, benzo5-6 membered heterocyclyl or heteroaryl; or optionally substituted with 1 or 2 R 8S1 and / or R 8S2 substituted 5-6 membered heterocycle or heteroaryl; or optionally substituted by 1 or 2 R 8S1 and / or R 8S2 substituted 5-6 membered heterocyclic or heteroaromatic ring and 5-6 membered heterocyclyl or heteroaryl, wherein R 8S1 selected from the group consisting of: -H, -OH, =0, halogen (-F, -CI, -Br, -I), -CN, -COOH; or C 1-10 alkyl, C 1-5 alkoxy, C 1-5 haloalkyl, C 1-5 alkanol, C 1-5 alkyl ester; or -C(O)NR e R f , C(O)NR e R f -(CH2) q -NR e R f , -SO2R g , -NR e R f -C 1-5 alkenoyl, wherein R e and R f are selected from -H, C 1-5 alkyl, and R g is selected from C 1-5 alkyl and amino, and R 8S2 selected from 5-6 membered saturated or partially unsaturated heterocyclyl, preferably R 8S2 selected from 17. The compound of claim 16, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R8is selected from: optionally substituted: 8S1 substituted: wherein 1 or 2 of U1-U5are each independently selected from 1 or 2 of N, O, and S, the remainder being carbon; 1 or 2 of V1-V6are each independently selected from N or O, the remainder being carbon; or R8is selected from: optionally substituted 8S1 or R 8S2 substituted wherein Q is CH or N, T1and T2are independently one of N, S, O or C, T3and T4are independently one of N, S, O or C, n is 1 or 2, wherein, denotes that a single, double bond or the ring is aromatic between the adjacent atoms of the ring.

18. The compound of claim 17, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R8is selected from:

19. The compound of any one of claims 13-18, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Ring A represents:

20. The compound of any one of claims 1-19, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein the compound is represented by Formula II: wherein ring A, L, R2, R3, R4, R5are defined as above, and ring B represents a saturated or partially unsaturated 5-6 membered heterocyclic ring comprising 1 or 2 heteroatoms selected from O, N and S.

21. The compound of claim 20, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: Compounds of Formula II have the structure shown below: wherein R9is selected from: -H, halo (e.g., -F, -CI, -Br, -I), C 1-5 haloalkyl, C 1-5 alkanol, optionally substituted phenyl, or C 1-5 alkylene-C(O)NR 9a R 9b wherein R 9a and R 9b are selected from -H, C 1-5 alkyl.

22. The compound of claim 21, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R9 is selected from: halogens (e.g., -F, -Cl, -Br, -I), -CH3, -C3H7 (e.g., - i Pr), -C4H9 (e.g., - n Bu), -CH2CHF2, -CH2CH2OH, -CH2C(O)NMe2, -CH2C(O)NH2; or R9is selected from optionally substituted phenyl, wherein optionally substituted phenyl is substituted with one or more C 1-5 haloalkyl or halogen (e.g., -F, -CI, -Br, -I).

23. The compound of any one of claims 20 to 22, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R2is selected from the group consisting of: -H or halogen (e.g., -F, -CI, -Br, -I).

24. The compound of any one of claims 20 to 22, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R3and R4are each independently selected from the group consisting of: -H, -OH, halogen (e.g., -F, -Cl, -Br, -I), -O-C 1-5 alkanoyl, optionally, R3and R4are joined to form a heteroalkylene, wherein the heteroatom is selected from O or N, for example, the group formed by the joining of R4to R3is selected from 25. The compound of any one of claims 20 to 22, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R5is selected from: -H, -OH, halogen (e.g., -F, -CI, -Br, -I); optionally substituted C 3-10 cycloalkyl, optionally substituted C 4-10 cycloalkenyl, optionally substituted C 3-5 alkenyl; optionally substituted 5-6 membered saturated or partially unsaturated heterocyclyl or heteroaryl, wherein the heterocyclyl or heteroaryl comprises one or more heteroatoms selected from O, N, or S, and when comprising multiple heteroatoms, the heteroatoms are the same or different.

26. The compound of claim 25, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R5is selected from the group consisting of: optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted cycloheptyl, optionally substituted cyclooctyl, optionally substituted cyclononyl, optionally substituted cyclodecyl, optionally substituted cyclobutenyl, optionally substituted cyclopentenyl, optionally substituted cyclohexenyl, optionally substituted cycloheptenyl, optionally substituted cyclooctenyl, optionally substituted cyclononanenyl, optionally substituted cyclodecanenyl, optionally substituted propenyl, optionally substituted butenyl, optionally substituted pentenyl; or R5is selected from the group consisting of: optionally substituted thienyl, optionally substituted thiazolyl, optionally substituted pyridyl, optionally substituted morpholinyl, optionally substituted piperazinyl.

27. The compound of claim 26, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein: R5 is selected from: -C3H5 (e.g., isopropenyl), cyclopropyl, cyclohexenyl substituted with -OH (e.g., ), 28. The compound of claim 1, or a pharmaceutically acceptable salt or stereoisomer, isotopic derivative, or prodrug thereof, wherein, Compounds represented by Formula I are selected from the group consisting of the compounds in the following table:

29. A pharmaceutical composition comprising a compound according to any one of claims 1-28, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, or prodrug thereof, and a pharmaceutically acceptable carrier or excipient.

30. Use of a compound according to any one of claims 1-28, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, or prodrug thereof, in the manufacture of a medicament or kit for reducing the activity of WRN in a cell or in a subject in need thereof.

31. The use according to claim 30, wherein the cell is a cell derived from a proliferative disease, preferably the cell is a cancer cell, preferably the cell is an MSI cancer cell.

32. The use according to claim 31, wherein the subject has a proliferative disease, preferably the proliferative disease is a cancer, preferably the cancer is an MSI cancer.

33. The use according to claim 32, wherein the MSI cancer is a Low-frequently microsatellite instability (MSI-L) cancer or a High-frequently microsatellite instability-high (MSI-H) cancer.

34. The use according to claim 32, wherein the MSI cancer is characterized by high microsatellite instability.

35. The use according to claim 32, wherein the MSI cancer is selected from the group consisting of colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, skin cancer, adrenal cortex cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and / or prostate cancer.

36. The use according to claim 30, wherein the subject is a mammal, preferably a human or a veterinary animal.

37. A method of reducing the activity of WRN in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-28, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, or prodrug thereof, or a pharmaceutical composition according to claim 29.

38. A method of treating a proliferative disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1-28, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, or prodrug thereof, or a pharmaceutical composition according to claim 29.

39. A method of reducing the activity of WRN in an MSI cancer cell, the method comprising contacting the cell with a compound according to any one of claims 1-28, or a pharmaceutically acceptable salt, stereoisomer, isotopic derivative, or prodrug thereof, or a pharmaceutical composition according to claim 29.

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