Camptothecin derivative for treating or preventing cancer, and antibody-drug conjugate thereof

Abstract

The invention relates to a camptothecin derivative for treating or preventing cancers, a drug linker conjugate and an antibody drug conjugate thereof, or a pharmaceutically acceptable salt or ester, a solvate, a tautomer, a stereoisomer, a prodrug or an isotope label thereof. The invention also relates to a pharmaceutical composition containing the camptothecin derivative or the drug linker conjugate or the antibody drug conjugate thereof, and a preparation method and application of the camptothecin derivative or the drug linker conjugate or the antibody drug conjugate.

Description

(12) International application published under the Patent Cooperation Treaty (19) wer Pe ZZ LNA AMO AT oe = (10) International Publication Number (43) International Publication Date 21WO 2024 / 255740 Al 2024 4F December 19 9 (19.12.2024) WIPO!|PCT 6D International Patent Classification: IR, IS, IT, JM, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, CO7D 491 / 22 (2006.01) A61LK 31 / 541 (2006.01) LA, LC, LK, LR, LS, LU, LY, MA, MD, MG, MK, MN, A6I1K 47 / 68 (2017.01) A61P 35 / 00 (2006.01) MU, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, CD International Application Number: PCT / CN2024 / 098491 PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, (22) International Application Date: 2024 4£ 6 A 11 A (11.06.2024) UA, UG, US, UZ, VC, VN, WS, ZA, ZM, ZW. (25) Application Language: Chinese (84) Designated Country (Unless otherwise specified, each available region is required) (26) Language of publication: Chinese protected): ARIPO (BW, CV, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SC, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasia (30) Priority: (AM, AZ, BY, KG, KZ, RU, TJ, TM), Europe (AL, AT, BE, 202310696385.9 2023466 A134 (13.06.2023) CN BG, CH, CY, CZ, DE, DK. EE, ES, Fl. FR, GB, GR. HR. 2023 11768933.0 December 21, 202321.12.2023) CN HU, IE, IS, IT, LT, LU, LV, MC, ME, MK, MT, NL, NO, (7D) Applicant: Chengdu Brilliant Pharmaceutical Co., Ltd. PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, (CHENGDU BRILLIANT PHARMACEUTICAL CO., CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, LTD.) [CN / CN]; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). TD, TO). (72) CHEN, Tonghun; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). CAI, Jun; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN), WANG, Tong; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). FANG, Xuerong; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). LIAO, Jianyu; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). HUANG, Haoxi; No. 263, Hexiang Third Street, High-tech Zone, Chengdu, Sichuan Province, China, Sichuan 610094 (CN). (74) Agent: Zhongzi Law Office; 7th Floor, New Era Building, No. 26, Ping'anli West Street, Xicheng District, Beijing, China,Beijing 100034 (CN). II (81) As the designated country otherwise specifies, claiming national protection for each available species): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, — BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, — CV, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, — GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IQ, — (54) Title: CAMPTOTHECIN DERIVATIVE FOR TREATING OR PREVENTING CANCER, AND ANTIBODY-DRUG CONJU- - GATE THEREOF (G4) Invention Title: Camptothecin derivative for treating or preventing cancer, and a drug-linker conjugate and antibody-drug conjugate thereof, or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug or isotope marker thereof. The present invention also relates to a pharmaceutical composition containing the camptothecin derivative or the drug-linker conjugate or antibody-drug conjugate thereof, and a preparation method therefor and a use thereof.PO a A, a ECT 25 To use salts or esters, solvates, tautomers, stereoisomers, prodrugs or isotope-labeled substances. Also relates to pharmaceutical compositions comprising the said camptothecin derivative or its drug linker conjugates or antibody-drug conjugates, methods of their preparation, and uses. II WO 2024 / 255740 PCT / CN2024 / 098491 FATA BOT TE RAT ED Re th RD Technical Field 5 In general, the present invention relates to camptothecin derivatives for the treatment or prevention of cancer, and their drug linker conjugates and antibody-drug conjugates, or their pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs or isotope-labeled substances. The present invention also relates to pharmaceutical compositions comprising the said camptothecin derivative or its drug linker conjugates or antibody-drug conjugates, methods of their preparation, and uses. 10. Background of the Invention: Antibody-drug conjugates (ADCs) are a promising type of drug, mainly composed of three parts: an antibody (Ab) responsible for selectively recognizing tumor cell surface antigens, a small molecule cytotoxic drug (Payload) responsible for killing tumor cells, and a linker (Linkem) that links the Payload to the Ab. The antibody acts as a carrier to target and deliver the small molecule cytotoxic drug to the target cells. 15. Through the binding of the antibody to the tumor cell surface antigen, the ADC drug enters the cell via endocytosis. Then, the small molecule cytotoxic drug dissociates from the ADC and exerts its biological function. Therefore, ADCs combine the powerful killing effect of traditional small molecule cytotoxic drugs with the tumor-targeting properties of antibodies. Currently, many ADC drugs are used for the treatment of tumors and solid tumors, such as POLIVY (Polatuzumab).Vedotin-piig (CD79b-MMAE) is FDA-approved for the treatment of relapsed or refractory diffuse large B4H BLCL (R / RDLBCL); PADCEV (Enfortumab vedotin-ejfv, 20"Nectin4-MMAE) is FDA-approved for the treatment of locally advanced or metastatic urothelial carcinoma (UC); Enhertu (Fam-trastuzumab deruxtecan-nxki, Her2-Dxd) is used to treat breast cancer, etc.; and Trodelvy (sacituzumab govitecan-hziy, TROP2-SN38) is used to treat patients with metastatic triple-negative breast cancer. Camptothecin (CPT) is a small-molecule plant antitumor drug used as a DNA topoisomerase inhibitor, showing good efficacy against intestinal, bone, and head and neck cancers. Several camptothecin derivatives have been developed, such as irinotecan 25. "(Irinotecan) and exatecam, they have been used clinically to treat tumors, including colorectal cancer, pancreatic cancer and lung cancer. Camptothecin derivatives have also been used in the development of ADCs, such as Enhertu which uses Dxd as payload and Trodelvy which uses SN-38 as payload. However, many solid tumors still respond poorly to camptothecin drugs (Joshua Z Drago, Shanu Modi, Sarat Chandarlapaty et al. Unlocking the potential of antibody-drug conjugates for cancer therapy. Nat Rev Clin Oncol. 2021 Jun;18(6): 327-344. dot: 30 ~~: 10.1038 / s41571-021-00470-8. Epub 2021 Feb 8.). Tumor cells can respond to camptothecin drugs through various mechanisms, such as reducing the accumulation of drugs in cells.Drug resistance to drugs and their ADCs (GL Beretta, L Gatti, P Perego et al. Camptothecin resistance in cancer: insights into the molecular mechanisms of a DNA-damaging drug. Curr Med Chem. 2013;20(12):1541-65. doi: 10.2174 / 0929867311320120006.) is related to x45 SN38 being a substrate of P-glycoprotein (Michael Tagen, Yanli Zhuang, Fan Zhang et al. P-glycoprotein, but not 35 multidrug resistance protein 4, plays a role in the systemic clearance of irinotecan and SN-38 in mice. Drug Metab Lett. 2010 Dec;4(4):195-201.). Therefore, there is still a need to develop new camptothecin derivatives with good cytotoxic effects and highly effective, low-toxicity ADC drugs. In particular, the camptothecin derivatives and their ADC drugs of the present invention have good therapeutic efficacy. Furthermore, the camptothecin derivatives and their ADC drugs of the present invention have good tolerability, few toxic side effects, and a wide therapeutic window. The ADC drugs of the present invention also have good tumor tissue targeting and a strong bystander effect. Summary of the Invention 5 Invention Overview In a first aspect, this application provides antibody-drug conjugates of formula (D) or pharmaceutically acceptable salts or esters thereof, solvates, tautomers, stereoisomers, prodrugs, or isotope-labeled compounds. In a second aspect, this application provides compounds of formula (D) or pharmaceutically acceptable salts or esters thereof, solvates, macromutators, stereoisomers, prodrugs, or isotope-labeled compounds. 10 In a third aspect, this application...The application provides drug linker conjugates of formula (D) or pharmaceutically acceptable salts or esters thereof, solvates, tautomers, stereoisomers, prodrugs, or isotopic labels thereof. In a fourth aspect, the application provides pharmaceutical compositions comprising the substance of the invention as defined herein, and pharmaceutically acceptable carriers, diluents, or excipients. In a fifth aspect, the application provides the substance of the invention as defined herein for the treatment or prevention of cancer. In a sixth aspect, the application provides a method for the prevention or treatment of cancer in a patient, the method comprising administering a therapeutically effective amount of the substance of the invention as defined herein to the patient. In a seventh aspect, the application provides the use of the substance of the invention as defined herein in the preparation of a medicament for the treatment or prevention of cancer. In an eighth aspect, the application provides a medicament for the treatment or prevention of cancer, the medicament comprising the substance of the invention as defined herein, and other common pharmaceuticals. The common pharmaceuticals may have the same or different effects as the substance of the invention. In a ninth aspect, the application provides a method for preparing the substance of the invention as defined herein. The substances of the invention include camptothecin derivatives of the invention and their drug linker conjugates and antibody-drug conjugates. It exhibits high cytotoxicity and can be used to treat or prevent tumors such as cancer, including but not limited to solid tumors, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver tumors, digestive system cancers (e.g., intestinal tumors, upper gastric cancer, gastric cardia cancer, esophageal cancer, appendiceal cancer, colon cancer, rectal tumors, colorectal cancer, pancreatic cancer), bladder tumors, melanoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, basal cell carcinoma, bile duct cancer, squamous cell carcinoma, thyroid cancer, brain cancer, head and neck cancer, and abdominal cancer.Membrane carcinoma, renal carcinoma, urothelioma, testicular carcinoma, central nervous system tumors (e.g., glioma, glioblastoma such as glioblastoma multiforme, glioma, or sarcoma), choriocarcinoma, oral epidermoid carcinoma, or hematologic malignancies, especially leukemia (e.g., acute or chronic diffuse myeloid leukemia, acute or chronic granulocytic leukemia), lymphoma (e.g., Hodgkin's lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), multiple osteomas. Preferably, the cancer is ovarian cancer, colon cancer, lung adenocarcinoma, or oral epidermoid carcinoma. The substance of the present invention has stronger antitumor activity compared to Dxd and SN38. Invention Details 35 fe, AS ARE TT DM a Ab Lyte te ted | (I) n 2 WO 2024 / 255740 PCT / CN2024 / 098491 BOA FAche. TANG, BAR. SKE MIAL HIZB IA Ase ily, wherein: Ab is an antibody or its antigen-binding segment; LI are each independently a linker unit; 5 Lo $B eae Ha e A PRA ERE IC: -(CRmRa)-(L)o-1-(CH2CH2O)5-(CRmRn)i-CO-, -(CRimnRa):-LM-(CRmRa):-(Lm)o-1-Lx-Lo-(CH2O)s-(CRmRn)-CO-; -(CRinRa)iL-(CRmRn)N(Rp)-(CRmRn)iN(Rp)-(CRmRa)-CO-, #H-(CRmRn)-Lu-CH(Rp)-CO-, HH, Ls' -CO- end with Lak, A Lo im Li eH, GFA where 10 Ra and Ran are each independently alkyl or Cl5 alkyl, LM are each independently -NH-CO- or -CO-NH-, LN & & Baz dt Fé -(CH2CH2O)m-(CH>)-. -(CH2)-(OCH2CH2)m-. -(3-8FEAR 2&)-(CH2) -(OCH2CH2)m BR-(OCH2CH2)m-(CH2):-(3-8 membered heteroarylene group)-, each Ri is independently -CO-(CHCHO)m-CH3 or -(CHz)rLw-(CH2CH2O)m-CH3, s and t are each independently 0, 1, 2, 3, 4, 5, 6, 7 or 8, and each m is independently 2, 3, 4, 5, 6, 7 or 8; each L3 is independently an amino acid residue, a short peptide chain composed of 2-10 amino acid residues, or -NH-(CH2)p-CO-; each La is independently a single bond or a spacer unit; each D is independently a small molecule drug moiety derived from a compound of formula GD; Ry Ri O N \ R2 Ho (II) wherein, Ri is H; CN; OH; SH; C1-6 alkyl; COOH; COOR; alkyl, wherein each alkyl group is optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and amino; -(CH2)p-(C3-8 cycloalkyl) or -(CH2)p-(C3-8 heterocycloalkyl), wherein said cycloalkyl and heterocycloalkyl are each optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH, C1-6 alkyl, C1-6 hydroxyalkyl, C1-6 cyanoalkyl, and C1-6 aminoalkyl; -(CH2)-(C1-6 alkoxy), -(CH2)-(C1-6 alkoxy); -(CH2)b-NRaRb; -(CH2)p-CH=NR; -NH-CO-R; -NH-CO-(CH2)-CH(OH)-Ra; -NH-(CH2)p-(CH=CH)-Ra; or -COOH; R2 R3 H; NO2; C1-6 alkyl; C1-6 alkoxy; C1-6 haloalkyl; C1-6 -NH-CO-(C1-6 alkyl); or NRdRe; Ro and Rs together with the atoms to which they are attached form4 R4 is R or Cg cycloalkyl, preferably Hi; Rs and Re are each independently Hy Cro ESE, -CO-(Cr6 HESE), -SO-(C1-6 ESE), -SO2-(C1- alkyl), 3 WO 2024 / 255740 PCT / CN2024 / 098491 BY-(CH2)q-(C3-8 cycloalkyl), said alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and毛基, and said cycloalkyl is further optionally substituted with one or more substituents independently selected from Cl6 alkyl, Ci-s PAGE, Cis FEB, Cis ERA, Cie RECN, RARER, and p and q are each independently 0, 1, 2, 3, 4, 5 or 6; 5 wherein the attachment point of D to L4 is at Ri BY Rs beet dD Fa aE, nn is an integer from 0 to 10, preferably an integer from 0 to 8. In another aspect, the present application provides an antibody-drug conjugate of formula (I) Ab-L1-Lo-L3-La-D (I) n 10 or a pharmaceutically acceptable salt or ester, solvate, tautomer, stereoisomer, prodrug or isotope derivative thereof, wherein Ab is an antibody or an antigen-binding fragment thereof; each LI is independently a linker unit; each Lo is independently a linking unit of the formula -(CHCHO)x-(CRaRuy)CO-, wherein the -CO- end is connected to L3, and the other end is connected to L1, and wherein Ra and Rn are each independently H or Cio ZE, x is 0 to 5, y is an integer from 1 to 5, and x+y<6; each L3 is independently an amino acid residue, a short peptide chain consisting of 2-10 amino acid residues, or -NH-(CH2)p-CO-; each La is independently a single bond or a spacer unit; each D is independently a small molecule drug moiety derived from a compound of formula (II); Ry Ri O N \ 4 %o Re HO 二 _O 20 (II) theIn this context, Ri is: H; BA: CN: OH; SH: Cyc HEFE. C6 MeFR EK Coo RSE, Pribbese. Mase AIA. Each of the following groups is optionally substituted by one or more substituents independently selected from halogens, CN, OH, SH, and amino groups; -(CH2)p-(C-cycloalkyl)] or -(CH2)p-(C3-s heterocycloalkyl), wherein the cycloalkyl and heterocycloalkyl groups are optionally substituted by one or more substituents independently selected from 25. Big. CN. OH, SH, 28. C6 GEFE. Crs de-alkyl, C16 alkyl, Ci-cyanoalkyl, and Ci-6 aminoalkyl: -(CH2)-(C1s alkoxy), -(CH)-(Ci-6 CHEE); -(CH2)b-NRaRb; -(CH2)p-CH=NR; -NH-CO-R; -NH-CO-(CH2))-CH(OH)-Ra; -NH-(CH2)p-(CH=CH)-R,; or -COOH:; Ro fll Rs & Aha A IA. HEL NOo. Crs GER. Cre BITRE. Cro PASE GEES Cre 30 eR. -NH-CO-(C1-6 F23E GFE). BK NRARD, BO Ro and Rs together with the atoms they are attached to form 4 RaA H BK C33 AGE, FLYEN H; A WO 2024 / 255740 PCT / CN2024 / 098491 Rs and Rb are each independently Hy C6 HERE. -CO-(Ci-6 KEE). -SO-(C1-6 HEE). -SO2-(C1-6 CFE). BY-(CH2)q-(C3-8 cycloalkyl), The alkyl and cycloalkyl groups are each optionally substituted with one or more substituents independently selected from halogens, CN, OH, SH, and cycloalkyl, and the cycloalkyl group is further optionally substituted with one or more substituents independently selected from Cl6 alkyl, Ci-s PAGER, Cig FEF EAE, Cis EERE Cl aminoalkyl; and 5p and q are each independently substituted with one or more substituents.0, 1, 2, 3, 4, 5, or 6; wherein the linking point of 5 Ly is on Ri or Rs or linked by the oxidative oxygen shown in formula (GD), and nn is an integer from 0 to 10, preferably an integer from 0 to 8. In another aspect, the present invention provides an antibody-drug conjugate with the formula: Ab Lyte tet | (I) 10 n BA AER OAR. TG. BARR. SEE A. HIZO BRIA ASR ily, wherein: Ab is an antibody or its antigen-binding fragment; LI are each independently a linker unit; 15 Ly are each independently a linker unit of the formula -(CHCHO)x-(CRaRaoyrCO-), wherein the -CO- end is connected to L; HE. A is connected to 1 at one end, and wherein Rn and Rn are each independently integers from HOBR Cis ESE, x HO BS ARM, yl to $, And x+y<6; L3 are each independently an amino acid residue, a short peptide chain composed of 2-10 amino acid residues, or -NH-(CH2)p-CO-; L4 are each independently a valence bond or spacer unit; 20 D are each independently a small molecule drug moiety derived from the formula (GD compound; Ry Ni ON \ 4 %o Re HO =O (II) where, Ri is, H; pies: CN: OH: SH: Ci-6 bese. Coo MFR Coo RAE, Prk. Mace A SLE A EER ak Se a A PR. CN. OH. SH FIREMEN; -(CH2)p-(C3- 25 ANGER), PRIA Gee (Eee — PRS Mi hit AR. CN. OH, SH #IZAZE RL RERL 4&3 -(CH2)p-(Cis GEA); (CH2)p(Cis Alkylthio), -(CH)p-NRaRo, -CH=NRa; -NH-CO-Ra:; -NH-CO-(CH2)p-CH(OH)-Ra: 2%-COOH; Ro fl R3 4 AoA pA. GE. NOo. Crs GES. Crs PATRUESES CrsPEGE. Cr alkoxy, or NRaRe, or Rs and Rs SEPT Be RF ET mk 4 8-membered heterocyclic group; 30 R4aA H BK C33 AGE, FLYEN H; Ra fll Re are each independently alkyl, -SO-(Cl6 alkyl), -SO-(Ci5 alkyl), or -(CH2)q-(C3s cycloalkyl), wherein the alkyl and cycloalkyl groups are each optionally substituted by one or more substituents independently selected from alkyl, CN, OH, SH Fa 5 WO 2024 / 255740 PCT / CN2024 / 098491; p and gq are each independently 0, 1, 2, 3, 4, 5 or 6; FP DS Ly ARES AE Ri or Rs are linked to or through the hydroxyl oxygen shown in formula (GD), and n is an integer from 0 to 10, Preferably an integer from 0 to 8. 5. In another aspect, the present invention provides camptothecin derivatives of formula (GD; Ry Ri ON \ 4 O R2 HO ne) (II) or pharmaceutically acceptable salts or esters, solvates, tautomers, stereoisomers, prodrugs or isotopic labels thereof, wherein 10 Ri is: H; eb: CN: OH: SH;, C15 alkyl, C: 5 alkenyl or Co RSE, Arse, Hse AI group each optionally substituted by one or more substituents independently selected from alkyl, CN, OH, SH and nitrogen groups; -(CH2)p-(C3-Th fot Sek) BK -(CH2)p-(C3-8 heterocyclic alkyl), said cycloalkyl and heterocyclic alkyl each optionally substituted by one or more substituents independently selected from Bi, CN, OH, SH, S38, Ci6 GEE, Cro EEF RGESE, Cro FESR GEE. Cro USE KESE. A Ci aminoalkyl substituents: -(CHo)p-(Cio KEE); = -(CHd)p-(Ci-6 alkylyl); -(CH2)p-NRaRb : 15 -(CH»)p-CH=NR,;-NH-CO-Ry; -NH-CO-(CH2)n-CH(OH)-Ra; -NH-(CH2)p-(CH=CH)-Ra; R1 is H, halogen, C1-10 alkyl, C1-5 hydroxyalkyl, C1-6 alkoxy, -NH-CO-(C1-6 hydroxyalkyl), or NRaRb, or R1 and R3 together with the atoms to which they are attached form a 4- to 8-membered heterocyclic group; R4 is H or C3-8 cycloalkyl, preferably H; Ra and Rb are each independently H, C1-5 alkyl, -CO-(C1-5 alkyl), -SO-(C1-5 alkyl), -SO2-(C1-5 alkyl), or -(CH2)q-(C3-8 cycloalkyl), wherein said alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and amino, and said cycloalkyl is further optionally substituted with one or more substituents independently selected from C1-6 alkyl, C1-6 alkoxy and C1-6 aminoalkyl, and p and q are each independently 0, 1, 2, 3, 4, 5 or 6. In another aspect, the present invention provides a compound of formula II: wherein the groups are each optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and amino; -(CH2)v-(C3-8 cycloalkyl), wherein said cycloalkyl is optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and amino, -(CH2)p-(C1-6 alkoxy); -(CH2)p-(C1-6 alkylthio), -(CH2)p-NRaRb, -CH=NRa, -NH-CO-Ry-NH-CO-(CH2)n-CH(OH)-Ra; or -COOH; R0, R1 and R3 are each independently H, C1-16 alkoxy, or NR5R6, or R0 and R5 together with the atoms to which they are attached form a 4- to 8-membered heterocyclic group; R4 is H or C3-8 cycloalkyl, preferably H; Ra and Rb are each independently H, C1-6 alkyl, -SO-(C1-6 alkyl), -SO2-(C1-6 alkyl), or -(CH2)q-(C3-8 cycloalkyl), wherein said alkyl and cycloalkyl are each optionally substituted with one or more substituents independently selected from halogen, CN, OH, SH and amino, and p and q are each independently 0, 1, 2, 3, 4, 5 or 6. In another aspect, the present invention provides a drug-linker conjugate of formula (I): Lg-L1-L2-L3-L4-D (III), wherein L1, L2, L3, L4 and D are as defined herein; and wherein Lg and L1 together form a leaving group; and Lg is F-, Cl-, Br-, CF3SO3-, MesN+ or a diazo group. More preferably, Lg is F, Cl, Br, or methanesulfonyl. Particularly preferably, Lg is methanesulfonyl. In some embodiments, R is: H; F; CN; OH; SH, C1-5 alkyl; C1-6 haloalkyl; C1-6 hydroxyalkyl; -(CH2)p-(C3-8 cycloalkyl) or -(CH2)p-(C3-8 heterocycloalkyl), which is optionally substituted with OH, SH, C1-6 alkyl and C1-6 carboxyalkylSubstituents: -(CH2)p-(C1-6 alkoxy); -(CH2)p-(Cis alkyl FE ) + -(CH)rNRaR ; -(CH)rCH=NR. ;:, -NH-CO-R, : -NH-CO-(CH2)p-CH(OH)-Ra: -NH-(CH2)p-(CH=CH)-R.:; #%-COOH. #E—2Fuh, Ra Al Re A is independently H, Cl6 alkyl, Cl6 hydroxyalkyl, Ci5 nitrogen alkyl, -SO:-(Cl6 alkyl), or -(CH2z)d- (optionally substituted with one or more Cs cycloalkyl groups independently selected from hydroxyl and Cl hydroxyalkyl). In other embodiments, Ri is H; Cr human alkyl:Cl6 alkyl:, Cl hydroxyalkyl:, -(CH2)p-(C3-IBGE); -(CH2)j-(C3s hydroxycycloalkyl); -(CH2)p-(C3-8 heterocycloalkyl), said heterocycloalkyl is optionally substituted with one or more groups independently selected from OH, Cl5 human alkyl and Cl The hydroxyl group is substituented with an alkyl group, preferably selected from OH and Cl-hydroxyalkanes, more preferably from Cl-6 hydroxyalkyl groups; -(CH2)p-(Ci-s alkyloxy); -(CH2)p-(Ci-6 alkylthio); 7 WO 2024 / 255740 PCT / CN2024 / 098491 -(CH»)p-NRaRp; -(CH2)p-CH=NR.; -NH-CO-R,; -NH-CO-(CH2))-CH(OH)-R,; -NH-(CH2)p-(CH=CH)-Ra; or -COOH. Further, Rs and Ru are each independently Hy Cro HEEL Cis FETE GEE. Cio BR. -SO2-(C1-6 HEE). -(CH2)o-(C3-3 ACE). -(CH2)q-(C3-8 AE tS). -(CH2)4-(C33 AGE substituted with one or more nitrogen groups). -(CH2)o-HE-P MET Cio hydroxyalkyl substituted with 5 Css cycloalkyl). In some other embodiments, RiWH: Creo alkyl:, Cro BITRE: Cro FESR GEE; © -(CH2)p-(C3-2 heterocyclic alkyl), said heterocyclic alkyl is optionally substituented by one or more substituents independently selected from OH and Ci hydroxyalkyl, more preferably -(CH2)p-NH(Ci-6 alkyl); -(CH)p-NHCI hydroxyalkyl); -(CH2)p-NH(Ci-6 nitrogen alkyl): 10 ""-(CH2z)b-NH-(CH2)a-(Cs-s cycloalkyl optionally substituted by one or more Cl- hydroxyalkyl): -(CH2z)pN(Ci-6 alkyl)(-SO2-Cl6 alkyl); -(CH2)p-CH=N(C1-6 HEAR); -NH-CO-(Ci-5 hydroxyalkyl); -NH-CO-(C3-s 258 It HE); -NH-CO-(CHz)b-CH(OH)-(Cls alkyl); -NH-CO-(CHz)p-CHOH)-(Css cycloalkyl), -NH-(CH>)p-(CH=CH)-(Cie alkyl): or -COOH. In other embodiments, Ri AH: Cro WBE: Cro alkyl:, Cro hydroxyalkyl: -(CH2)p-(C3-15" heterocyclic alkyl), wherein the heterocyclic alkyl is optionally one or more Cro FESR GEREN s -(CH2z)p-(Cis alkoxy), -(CH2)n-(Ci65 alkathio); -(CH2)p-NHo; -(CH2)p-NH(Ci-6 alkyl):; -(CH)-NH(Cis hydroxyalkyl); -(CH2)p-NH(C1-6 aminoalkyl); -(CH)p-NH-(CH2)a-(C3-s cycloalkyl), wherein the cycloalkyl is optionally substituted by one or more substituents selected from Cro hydroxyalkyl; -(CH>)pN(Ci-6 alkyl)CSO-Ci6 ERE); -CH=N(Ci-ese); -NH-CO-~(C1-6 hydroxyalkyl); -NH-CO-(C3.3 #2E 4 bid); -NH-CO-(CH)p-CH(OH)-(Ci-alkyl); 20.-NH-CO-(CH2z)-CHCOH)-(C3s cycloalkyl); -NH-(CH2)p-(CH=CH)-(Ci5 hydroxyalkyl); or -COOH. In other embodiments, R; is Hs Cio alkyl: Cu alkyl: Cio hydroxyalkyl:, -(CH2)p-(Cs3-heterocyclic alkyl), wherein the heterocyclic alkyl is optionally substituted with one or more Cro hydroxyalkyl; -(CHa)p-(Ci-o Seid); -(CH2)p-NH, -(CH2)p-NHCCI5 alkyl); -(CH2)p-NH(C1-6 FEE ESE); -(CH2)p-NH(C1-6 REE); -(CH2)p-NH-(CH2)q-(C3s Atk), wherein the cycloalkyl is optionally substituted with one or more substituents selected from Ci hydroxyalkyl for 25" substituents:, -(CH2)pN(Ci6 alkyl)(-SO-Ci5 HERE); -CH=N(Ci-6 alkyl), -NH-CO-(Ci6 carboxylalkyl), -NH-CO-(C3-3 Hydroxyl cycloalkyl); -NH-CO-(CH2)p-CH(OH)-(C3-3 cycloalkyl); -NH-(CH2)p-(CH=CH)-(C1-6 hydroxyalkyl); or -COOH. In some embodiments, RI WH: Cl6 alkyl: -(CH2)s-(C3-3 AGERE optionally substituted with one or more Cl hydroxyalkyl): -(CH2)-(Cl6 alkylthio); -(CH2)p-(NH2); -(CH2)p-NH(C1-6 663%); -(CH2)p-NH(C1-6 30" alkylalkyl); -(CH2z)jp-NH-(CH2)a-(C1-6 663%) optionally substituted with one or more Cl hydroxyalkyl); -NH-CO-(CH2)p-CH(OH)-(C3-8 cycloalkyl); EK-NH-(CH2)p-(CH=CH)-(C1-6 alkyl). In some embodiments, RI is an organic, C16 alkyl, -(CH2)s- (optionally a C1-6 nitrogen-containing heterocyclic alkyl group substituted with one or more C15 hydroxyalkyl groups), -(CH2)p-(C1-6 alkylthioalkyl group).-NH2, -(CH)-NHCCi-5 alkyl, -NH(Ci16 hydroxyalkyl), -(CH2)p-NH-(CH2)q- (optionally a C38 cycloalkyl substituted with one or more Cro hydroxyalkyl), 35. -NH-CO-CH(OH)- (C3-8 cycloalkyl), or -NH-(CH=CH)- (Cu6 alkyl). In further embodiments, RiI is H, Cl6 alkyl, C3s substituted with one or more Ci hydroxy alkyl groups, -CH2~(C 1-6 ef). -NH2. -CH>-NH(C1-6 Wt2&). -NH(C1-6 236 Gt3e). -NH-({£ 8 WO 2024 / 255740 PCT / CN2024 / 098491 -NH-CO-CH(OH)-(C3-s G28). BK-NH-(CH=CH)-(C 1-6 #38). In some embodiments, Ri is Cl6 alkyl, -(CH2z)p-(optionally substituted with one or more C1 PEGE C3. RAY GER). -(CH2)p-(Ci6 alkylthio), -(CH2)-NH(Ci6 6638). -(CH2)p-NH-(CH2)q-CEXE 8 — one or more Css AG HE substituted with a Cro hydroxyalkyl group. Be -(CH2)p-NH (Ci-o hydroxyalkyl), or 5-NH-(CH>)p-(CH=CH)-(C1-6 alkyl). Preferably, Ri is Cro alkyl, -(CH2z)- (optionally a Cs-s cycloalkyl group substituted with a Ci hydroxyalkyl group), -CH2-(Ci6 alkylthio), -NHCCi-Gt. -NH (Ci-6 hydroxyalkyl), -(CH2z)p-NH-(CH2)u- (optionally a Cs3-s cycloalkyl group substituted with a Cl hydroxyalkyl group), or -NH-(CH=CH)-(Ci-alkyl). In some embodiments, Ri is a Cl6 alkyl group, -(CH2)p- (optionally substituted with a Cl-hydroxy alkyl group), -(CHo)p-(C1-6 RE), or -NH2.-(CH>2)p-NH(C1-6 Ste). -(CH2)p-NH(C1- P25 ERE). -(CHz)p-NH-(CH2)a-(optionally replaced by a Cro FEAR GEAR AUR C3-s GEE). -NH-CO-(C16 F228 S28). -NH-CO-(CH?2)»-CH(OH)-(C3-s cycloalkyl), or -NH-(CH=CH)-(Ctu6 alkyl). Preferably, R is a C1-alkyl, a C3-hydroxyalkyl substituted with a C1-hydroxyalkyl, -NH, -(CHz)p-NH (C1-6 alkyl), -(CHz)p-NH (C1-hydroxyalkyl), -NH- (C1-hydroxyalkyl substituted with a C1-hydroxyalkyl), -NH-CO- (C1-hydroxyalkyl), or 15-NH-(CH=CH)-(C1-6). In some embodiments, Ri MEA: H; RSE. ZI. ASE. JHE. ese. GSE, and chloromethyl, fluoroHAZE, UAE, Ale, ZR. ROS, BAZ. UAE. RAE. TSE. SR. OR butyl; FOSk SR, FESR CE, ESE. Fede SR. FESR IR. FESR CE, -(piperyl)-CH2OH: ~CHo-( Fig.). -CHo-(Z. ie). -CH2-(A i). -CHo-( J id&). -CHo-(R mE). -CH-~( Offi 20 4); -NH2, -NHMe, -NHEt. -NHPr. -NHBu. -NHUX2e). -NH( GE); -NH G36 FSS). -NHG2 JE ZTE). NH GSE ASE). -NH GSE TE). -NH GSE RS). -NH FEE OE); -NH(nitromethyl), -NH(228 2.3%). -NH (2028 2). -NH(2028 JE). -NH (2028 eee). -NH( 22 3S); -CH2-NHMe, -CH-NHEt, -CH-NHPr, -CH-NHBu, -CH2-NH(base), -CH2-NH( (hexyl);, -CH2-NH-CH2-(cyclopropyl);-CH-NH-CH2- (cyclobutyl), -CH2-NH-CH2- (cycloalkyl), 25 -CH-NH-CH2- (cyclohexyl); -(CH2)2-N(Pr)(-SO2Me); -CH=NMe, -CH=NEt, -CH=NPr, -CH=NBnu, -CH=N (pentyl), -CH=N (hexyl); -NH-CO- hydroxymethyl, -NH-CO- hydroxyethyl, -NH-CO- propyl, -NH-CO- ¥% 3& J 36. -NH-CO-¥% 3£ 3. -NH-CO-## 3% GSE; -NH-CO- (hydroxycyclopropyl), -NH-CO- (hydroxycyclobutyl), -NH-CO- (hydroxycycloalkyl), -NH-CO- (hydroxycyclohexyl); -NH-CO-CH(OH)- ( Two groups), -NH-CO-CH(OH)-(cyclobutyl), -NH-CO-CH(OH)-(cyclobutyl), -NH-CO-CH(OH)-(cyclohexyl); 30 -NH-CO-CH>-CH(OH)(Me), -NH-CO-CH2-CH(OH)(Et), -NH-CO-CH)-CH(OH)(Pr), -NH-CO-CH>-CH(OH)(Bu), -NH-CO-CH-CH(OH)(pentyl), -NH-CO-CH2-CH(OH)(hexyl); -NH-(CH2)-(CH=CH)-(CH2OH); or -COOH. In some implementations, Ri is H; alkylating agents: MUSE, FASE TSE; -(piperidinyl)-CHOH; -CH2-(dithioH group); -NH, -NH(hydroxyl); -CH-NH(pentyl); -NH(hydroxybutyl); -NH(aminobutyl); Donon, 35 -CH2-NH-CH2-(# Ke); -(CH2)2-N(Pr)(-SO2Me); -CH=N(pentyl); + -NH-CO-(#% 3k AE), -NH-CO-(hydroxycyclohexyl); -NH-CO-CH(OH)-( 4 i 3); ~-NH-CO-CH2-CH(OH)(Me); -NH-(CH2)-(CH=CH)-(CH2OH); or -COOH. 9 WO 2024 / 255740 PCT / CN2024 / 098491Press. Me IRIES BE PS Pha Pa. CN. OH, SH MASINI FLEW R; (CH2)p-(C3-2 UAE), ATR Gee EK — TMS ST HE IA. CN. OH, SH and nitrogen substituents:, -(CH2)-(Ci-alkoxy);, -(CH2)p-(Ci6 efi HE); -(CH2)p-NRaRo; 5 -CH=NR,:; -NH-CO-R,; -NH-CO-(CH2))-CH(OH)-Ra:; or -COOH; wherein Rs and R are each independently H, C16 alkyl, -SO-(Ci6 alkyl), -SO:-(C16 alkyl), or -(CH2)a-(C3-s cycloalkyl), wherein the alkyl and cycloalkyl are each optionally substituted by one or more substituents independently selected from halogens, CN, OH, SH and amino. Alkyl groups: -(CH2)p-(C3-2 AGE); -(CH2)n-(C3-s Fe FRA EAE); -(CH2)p-(C1-6 CAE); -(CH2)p-(C16 10 eWRFE); ~-(CH>2)p-NRaRb; -CH=NR.; -NH-CO-R.; -NH-CO-(CH2)»-CH(OH)-Ra; #%-COOH. hie 8 BE) = -(CH>)p-(Ci-6 i Gi ZE ) ; -(CH)oNRaRo ; -CH=NR,. ; -NH-CO-R, ; -NH-CO-(CH2)p-CH(OH)-Ra: or -COOH; preferably wherein Rs and Ru are each independently Hy Cr HUF. Ci POSE ERE. Cro nitrogen-based alkyl, -SO-(Cl6 EA). -(CH2)q-(C3-3 cycloalkyl), or -(CH2)q-(C3 sg hydroxycycloalkyl). 15 In a further embodiment, Ri AH; Cio ERE: Cro alkyl group: Cio FASE GEE; -(CH2)p-(Cis tie); -(CH2)p-(Ci-6 iti); -(CH2)p-NH>; -(CH)-NHCC6 alkyl); -(CH2)p-NH(C1-0 #22 70 alkyl);-(CH2)p-NH(C 1-5 22 G28); (CH2)p-NH-(CH2)q-(C3-s FA G62); -(CH2)pN(C 1-6 S73)(-SO2-C 1-5 ht BE); -CH=N(Cis bi 28); -NH-CO-(Cis hydroxyalkyl);, -NH-CO-(C3.3 hydroxycycloalkyl); -NH-CO-(CH?2)»-CH(OH)-(C1-6 alkyl); -NH-CO-(CH2)»-CH(OH)-(C3-3 Aes); or -COOH. 20 In some embodiments, RiI is HY Cro alkyl, -(CH2)p-(Cis Ge fit BE), -(CH2)p-(NH2), -(CH2)p-NH (C1-6 alkyl), -(CH)-NHCCLe hydroxyalkyl, -(CH)p-NH-(CH2)-(C3s cycloalkyl), or -NH-CO-(CH2)-CH(OH)-(C3s cycloalkyl). In a further embodiment, Ri W Hy Cro ESR. (CHa) p-(C1-6 GG BREE). -NHb. -(CH2)p-NH(C1-6 Ide). -NH(Ci6 Fe 2E GEE). -(CH2)p-NH-(CH2)q-(C3-s FR GEES). BK-NH-CO-CH(OH)-(C3-3 cycloalkyl). In a further embodiment, R WHY C6 alkyl, -CH:-(Ci- GEMS). -NH2. -CHo-NH(C1-« alkyl), -NH(CI6 alkylyl), -CH:-NH-CH2-(C3-s cycloalkyl), or -NH-CO-CH(OH)-(Cs3-s cycloalkyl). In some embodiments, RI is Cl₂-alkyl, -(CH₂)-(C₅ alkylthio), -(CH₂)p-NH (C₅ alkyl), or -(CH₂)p-NH (C₁L₆ hydroxyalkyl). Preferably, RI is C₆ alkyl, -CH₂-(C₆ alkylthio), -NH (C₅ alkyl), or 30"-NH (C₁L₆ hydroxyalkyl). In some embodiments, RI is C₆ alkyl, -(CH₂)-(C₁L₆ alkylthio), or -(CH₂)-(C₁L₆ alkylthio).-NH2, -(CH2)-NHCCL5 alkyl), -(CH2)p-NH(C1-6 C2236 tz), -NH-CO-(C1-5 F228 KtAK), BK-NH-CO-(CH2)p-CH(OH)-(C3-s cycloalkyl). Preferably, Ri Cre bes. -NH2. -(CH2)p-NH(Ci-6 4€5E), -(CH2)p-NH(Ci-6 carboxyalkyl), or -NH-CO-(Ci6 hydroxyalkyl). 35 In some embodiments, Ri WN: Cre alkyl; -(CH2)-(C3s heterocyclic alkyl), said heterocyclic alkyl is optionally substituted by one or more substituents independently selected from OH, Cis alkyl or Crs hydroxyalkyl; -(CH2)p-NRsRbo, wherein Ra and Re are each independently H, Cl15 alkyl, Ci6 hydroxyalkyl, or -(CH2)a- (Cs cycloalkyl optionally substituted by one or more substituents independently selected from alkyl and Cro hydroxyalkyl); or -NH-(CH2z)p-(CH=CH)-Ra, wherein Ra is Cl hydroxyalkyl, and p is 0, 1, 2, 3 or 4. Preferably, Ri is: Cl alkyl, -(CH2)- (optionally a C3-s heterocyclic alkyl substituted with a Cu6 hydroxyalkyl); (CH2)p-NRaRv, HEF RaW H, H Ro 4 Cis ER GER. B-( CHa) g (HE optionally substituted with a Cro EDU Coe cycloalkyl); BK-NH-(CH2)p-(CH=CH)-Ra, FER RaW Cyc Pek alkyl, and p is 0, 1, 2, 3 or 4. 5 In other embodiments, Ri is Cie alkyl; -(CH2)p-(C3-s RIA KEEL), FTIR ZR EEE one or more Cro FEE GEEDUR -(CH2)p-NH(C1-6 Fe SE GEES); -(CH2z)p-NH-(CH2)a-(C3-s cycloalkyl), wherein the cycloalkyl is optionally substituted with one or more Cro FEE GEEDUR.One of the substituents selected from Cr hydroxyalkyl is substituted; BK-NH-(CH2)p-(CH=CH)-(C1-6 hydroxyalkyl). In other embodiments, RI is Cl EAE; -(CH2)p-(C3-2 cycloalkyl), wherein the cycloalkyl group is optionally substituted with one or more Cro-alkyl groups; -(CH2)p-(Ci-6 alkylthio); -(CH)p-NH; -(CH2)p-NH(Ci-6 alkyl); -(CH2)p-NH(C1-6 hydroxyalkyl); -(CH2)p-NH-(CH2)a-(C3-s cycloalkyl), wherein the cycloalkyl group is optionally substituted with one or more substituents selected from Cro-alkyl groups; -NH-CO-(C1-5 cycloalkyl); or -NH-CO-(CH2)p-CH(OHD-(Cs-cycloalkyl). In other embodiments, RI is Cl6 HEHE: -(CH2)-(C3-s cycloalkyl), wherein the heterocyclic alkyl group is optionally substituted with one or more Cl6 groups. Alkyl substitution: -(CH)p-NH; -(CH2)p-NH(C1-6 bid); -(CH2)p-NH(C1-0 #228 Wi); -(CH2)p-NH-(CH2)q-(C3-8 cycloalkyl), wherein the cycloalkyl is optionally replaced by one or more of the following: -NH-CO-(C1-6 #22 bed); BK-NH-CO-(CH2))-CH(OH)-(C3-s cycloalkyl). In other embodiments, RI is Cu6 ESE; -(CH2)-(C3s cycloalkyl), wherein the heterocyclic alkyl group is optionally substituted with one or more Cl6 hydroxyalkyl groups; -(CH2)p-NHCC46 hydroxyalkyl; -(CH2)p-NH-(CH2)q-(C3-8 cycloalkyl), wherein the cycloalkyl group is optionally substituted with one or more substituents selected from Cu hydroxyalkyl groups; or -NH-CO-(CH2)»-CH(OH)-(C3-3 cycloalkyl). In other embodiments, RI is Cro alkyl.-(CH2)-(C3s heterocyclic alkyl), wherein the heterocyclic alkyl group is optionally substituted with one or more Cl6 hydroxyalkyl groups; -(CH2)p-NHCCL5 hydroxyalkyl); or -(CH)p-NH-(CH2)d-(Css ried), wherein the cycloalkyl group is optionally substituted with one or more substituents selected from Cus hydroxyalkyl groups. In further embodiments, pbp is 0, 1, 2, 3, or 4. In some further embodiments, p is 0. In other further embodiments, p is 1. In other further embodiments, p is 2. In other further embodiments, p is 3. In other further embodiments, p is 4. Methyl, bromomethyl, oxyethyl, fluoroethyl, bromoethyl, and chloropropyl, fluorodimethyl, bromodimethyl, oxyethyl, fluoropropyl, bromoethyl, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl, -CH2-(methylthio), -CH2-(ethylthio), -CH2-(propylthio), -CH2-(butylthio), -CH2-(pentylthio), -CH2-(hexylthio); -NH2. -NHMe,. -NHEt, -NHPr. -NHBu, -NH( / %2%). -NH(CU38); -NHG#28'F 38). -NHG#2e 38). -NHG# JLSL). -NHCG43£ J 38). -NHG22E 228). -NH (SE USE); -NH( -NH (nitromethyl), -NH (nitroethyl), -NH (aminopropyl), -NH (aminobutyl), -NH (nitropentyl), -NH (aminohexyl); -CH-NHMe, -CH2-NHEt, -CH-NHPr, -CH-NHBu, -CH-NH (pentyl), -CH-NH (hexyl); -CH2-NH-CH2- (cyclopropyl), -CH-NH-CH- (cyclobutyl), -CH-NH-CH- (cycloalkyl), -CH-NH-CH- (cyclohexyl):-(CH»)2-N(Pr)(-SO2Me); -CH=NMe, -CH=NEt, -CH=NPr, -CH=NBu, -CH=N(pentyl), -CH=N(hexyl); -NH-CO-hydroxymethyl, -NH-CO-hydroxyethyl, -NH-CO-hydroxydimethyl, -NH-CO-hydroxybutyl, -NH-CO- 11 WO 2024 / 255740 PCT / CN2024 / 098491 Fez MR, -NH-CO-##38 8; -NH-CO-GtE WS). -NH-CO-G438 J 38). -NH-CO-(F# FLIP MAL). -NH-CO-G23kf GSE); -NH-CO-CH(OH)-(4 38). -NH-CO-CH(OH)-( J 38), -NH-CO-CH(OH)-( / &% #6). -NH-CO-CH(OH)-( # Gi 3£); -NH-CO-CH2-CH(OH)(Me) , -NH-CO-CH»-CH(OH)(Et) . | -NH-CO-CH2-CH(OH)(Pr) , -NH-CO-CH2-CH(OH)(Bu) , 5 | -NH-CO-CH>-CH(OH)( / K 3%). -NH-CO-CH)-CH(OH)(4&); or -COOH. In some implementations, Ri is H; Esk; SAE, POSE TSE, -CH2-(propylthio): -NH2; -NHI(pentyl); -CH-NH(pentyl); -NH(hydroxybutyl); -NH(nitrobutyl); -CH-NH-CH2-(cyclohexyl); -(CH2)2-N(Pr)(-SO2Me); -CH=N( / % 3%); -NH-CO-(## 3& FA SE); -NH-CO-(carboxycyclohexyl); -NH-CO-CH(OH)-(cyclopropyl): -NH-CO-CH2-CH(OH)(Me); or -COOH. 10. In some embodiments, Ri1 is -NH (hydroxymethyl), -NH (hydroxyethyl), -NH (hydroxypropyl), -NH (carboxybutyl), -NH (hydroxyethyl), -NH (hydroxyhexyl); -(azacyclopentane)-(CHOH), -(azacyclohexane)-(CHOH), -(R1 is -NH-cyclopentane-(CHOH), -NH-cyclohexane-(CH2OH), or -NH-cycloheptane-(CH2OH). Preferably, R1 is -NH (hydroxyethyl), -NH (hydroxypropyl), -NH (hydroxybutyl), -NH (hydroxyethyl), -NH (hydroxyethyl); -(heterocyclic alkyl)-(CHOH); -NH-cyclohexane-(CHOH); or -NH-(CH2)-(CH=CH)-(CH2OH). In some embodiments, R1 is: -NH (hydroxyethyl), -NH (hydroxypropyl), -NH (hydroxybutyl), -NH (hydroxyethyl); -(heterocyclic alkyl)-(CHOH); -NH-cyclohexane-(CHOH); or -NH-(CH2)-(CH=CH)-(CH2OH). In other embodiments, R1 is: -NH (hydroxyethyl), -CH (propylthio), -NH (pentyl), -NH (hydroxyyl), -CHo-NH (2). -NH-CO-#238 ASE. -NH-CO-CH(OH)-(4 Ade). -(AZEY Cbe)-G2 FE). BY-NH-(H4 chft)-F2 HA). In other embodiments, RI WeedE. -NH>. -NH(ESE TE). -CHo-NH(R2E). -NH-CO-#% 20 " ylmethyl, -NH-CO-CH(OH)-(cyclopropyl), -(azacyclohexane)-(hydroxymethyl), or -NH-(cyclohexane)-(hydroxymethyl). In other embodiments, Ri is alkyl, -NH (hydroxybutyl), -NH-CO-CH(OH)-(cyclopropyl), -(azacyclohexane)-(hydroxymethyl), or -NH-(cyclohexane)-(hydroxymethyl). In other embodiments, Ri is alkyl, -NH (hydroxybutyl), -( (-(cyclohexane)-(hydroxymethyl), or -NH-(cyclohexane)-(hydroxymethyl). 25 In some embodiments, R: APIA, BOR. A. IR. WH. AEHE DWT RH, Ro AK. In some embodiments, R3 is Cie ESE. Cio HEA. KR. UGE. -NH2. -NH-CO-(Ci-6 hydroxyalkyl)Rs is preferably methyl, ethyl, adimethyl, butyl, fluorine, chlorine, bromine, -NH, -NH-CO-(CH2OH), or NO. More particularly, R3 is methyl, bromine, -NH2, -NH-CO-(CH2OH), or NO. 30. In some embodiments, R3 is a halogen, alkyl, NO:, C16 alkyl, C16 alkyl-substituted, C16 carboxyalkyl, C16 alkoxy, or NRaRb, wherein Rs and Re are each independently C165 alkyl, -SO-(C1-5 alkyl), -SO:-(C1-6 alkyl), or -(CH2)q-(C3s cycloalkyl), wherein the alkyl and cycloalkyl are optionally substituted by one or more substituents independently selected from halogen, CN, OH, SH, and hydroxyl. In some embodiments, R3 ∈ Cio Wek. Cio OME. EPR. UE. -NH2 or NO. In a further 35” embodiment, R3 is Cr Hes. BPA. -NH> or NO. In particular, Rs is methyl, ethyl, dimethyl, butyl, gL. Sl. YR. -NH2 or NO:. More particularly, Rs is methyl, bromine, -NH2 or NO:. In some embodiments, Rs3 is Cl6 alkyl or NRaRb, wherein Res and Re are each independently HBL-CO-(C1-6Ht alkyl), wherein the alkyl group is optionally substituted with one or more OH groups. Further, Rs is Cl alkyl, -NH2 or 12 WO 2024 / 255740 PCT / CN2024 / 098491 -NH-CO-(CH2OH). More further, R3: is Cl6 alkyl or -NH-CO-(CH2OH). In some embodiments, R3 is Cl6 alkyl, For example, methyl groups. In some embodiments, Rz and Rs, together with the atoms they are attached to, form a 4 to 8-membered group containing N, O, or...a heterocyclic group having $ as a heteroatom, for example, dioxacyclopentene, for example, in the compound of Example 6 or 7. 5 In some embodiments, D is linked to L4 through a nitrogen or oxygen atom in Ri or Rs, or through the hydroxyl oxygen atom shown in formula (I), for example, linked through an amide bond, a N-methyl ether bond or a carbamate bond. In some embodiments, D is a small-molecule drug moiety obtained by removing one atom (e.g., an oxygen atom) or an atomic group from a compound of formula (I), for example, removing a hydrogen atom connected to a nitrogen or oxygen atom in Ri or Rs, or removing an oxygen atom from the hydroxyl group shown in formula (I). Specifically, D is selected from the following moieties: I Cl 6 2° ~ N N} Ww oN / °o — F (er) v NA? STI-A-4 F Bog 20 STI-A-6" 3° 4 “ N O N O (er) (er) AS / MY AN D STI-A' Hor / STI-A2' Ho 7 O o (er) ne O se N N \_ 4 vn (er) ? or ° F HO 2 0 — 9 STI-F-04' STI-E-08' 7 Ko 8’ O H3C | ON nN VL mp O F not C q ‘ Is STI-F-8" — HO* = (er) STL 一 9 oat 10’ » ae HN O == ON w , oO em eeOT F novo F Hotz OO STI-F-03¢-1' STI-F2' ~~ 13 WO 2024 / 255740 PCT / CN2024 / 098491 Ll’ 12’ (er) (er) af tN Nn’ \ | (er) OoN N \ | (er) F Ho7: =O F 全 — 二 STI-G1' STI-G03' —™ 13° SS 14 LL S (er) s Oo F 0 20 F HO 20 STI-G2-06c' STI-G2" + ay + 15 0 16 ; ae o HN’ “9 O N == ON i jf oO H3C y / H3C ay SN Ww 半 HO’ 2°F HOY: O F , — STID, STI-F 17 at 18’ “ fe) AL N O NH 9 = N = N H3C HC / ~o PAL . AAS F How: 0 F How: Oo STI-F6’ ™ STI-F7' > > O 19 20 o2 入 Tu / N oO 9 H3C y \ | Oo ara Na NN I 0 N 二 Station F HO 20 F Ho"= 0 STI-G4 STI-F13" > Tu 21 0 22 o” 人 人, fe) Y 口 N O H3C oY fo HIN 0 N == ON F HO 2 0 HC wv» [| © SIT-F12R' F HO 2 0 STI-F10" 14 WO 2024 / 255740 PCT / CN2024 / 098491 Pl’ OO P2’ O 〇 we 7 nae O ™ a SN NH O H3C a\ If bb = ON N : H3C o\ If D F HO :- 60 N STI-F5' ~ F Ho 20 STI-F8’ P3’ is | Ko N o NH O H3C AS / Do H3C aN / F HO“: 0 F HOw: “‘O STI-F14' All STI-F15" —™ PS’ do P6’ ye a O HN — N 人 SS N O HsC 二 3 X / \ I ~o D NA 7 / 、 STI-F17' ~ STI-F48' \ 0 P7, O == ON ie) HN | re) 了 v O F How: 0 | — ww STI-G5' fe Hee EP, Each is independently: O o O 2 2 N a N N N N N= n~ N@ N o. * Me nae N maw oe NTS | pie b O pee ee Be N=, more preferably, wherein position 1 is connected to Ab, and position 2 is connected to Lo. 5 TE HES ATR A, Lo are each independently a linking unit selected from the following formula: -(CH2)r(Lwo-l-(CH2CH2O)s-(CH2)rCO-,-(CH2)rLM-(CH2)r(Lo-l-LN-LM-(CH2O)s-(CH2)rCO-, -(CH)eLw(CHOeNGR-(CH2)-N(Rp)- (CH2)-CO-, #(CH2)-Lat-CH(Rp)-CO-, 15 WO 2024 / 255740 PCT / CN2024 / 098491 In some implementations, the -CO- end of L2 & Eph Hh FE -(CRmRa)-(Lm)o-1-(CH2CH2O) -(CRmRa)i-CO-» fi) UI -(CRmRn)-(CH2CH2O);-(CRmRn):-CO- BK -(CRmRn)-Lu-(CH2CH2O)s-(CRmRa)-CO , FLAP Le is connected to Ls, and Le IN —mn-S Li is connected. In a further embodiment, L2 is independently 5, -(CHz)r(Lo-(CHCHO)-(CHJrCO-), for example -(CH2)-(CH2CH2O), -(CH2)-CO- or -(CH2):-Lu-(CH2CH2O)5-(CH2),-CO-, the -CO- end of EFA Le is connected to 3, and the other end of 1 is connected to LI. In a further embodiment, L: is independently -(CHz)r(CHCH2O):-(CHz)rCO-, -(CHz)rCO-NH-(CH2CH2O)-(CHz)rCO-, or -(CH)rNH-CO-(CHCH2O)s-(CH2)rCO-, preferably -(CH2):-(CH2CH2O).-(CH2):-CO-. #&%-(CH2):-CO-NH-(CH2CH2O);-(CH2)-CO-, #:4F L2's -CO- 10 "" end is connected to L3 ER, A Lo's other end is connected to LI. In some embodiments, I2 is independently -(CRnRnrLv-(CRnRor(Loi-LNL-(CHO)-(CRnRorCO-), for example -(CRanRrLv-(CRaRarLN-Lv-(CHO)s-(CRnRnrCO-), or -(CRmRn)-L-(CRmRa)-Ly-Lx-Ly-(CH2O)<-(CRmRn)-CO-,» FAA Le's -CO- end is connected to Ls, and Le 15 "'''''''''''''''''''''''''''''''''''' '"'' '''''''''''''''' '"'''''''''''''''''''''' '"'''''''''''''''''''''''''' ...-(CH2)rLvM-(CH2)r-LN-LM-(CH2O)s-(CH2)rCO-, or -(CH2)-Lw-(CH2)rLM-LN-L-(CHO)-(CH)CO-, where the -CO- end is connected to L; and the other end of Le is connected to LI. In a further embodiment, L2: is independently -(CH2):-CO-NH-(CH)):-(3-8 methylheteroaryl)-(CH)r(OCHCH)m-NH-CO-(CH2O)-(CH2)rCO- (preferably 20. -(CH2)-CO-NH-(CH2)-(= Mz Wy 8 ) (CH2):-(OCH2CH2)m-NH-CO-(CH20)-(CH2):-CO-), -(CH»),-CO-NH-(CH»),-NH-CO-(CH2CH2O)m-(CH2)-NH-CO-(CH20)-(CH2)-CO-. BY -(CH2)-CO-NH-(CH2);-NH-CO-(CH2):-(OCH2CH?)m-NH-CO-(CH20)-(CH2):-CO-, FEF Ly of -CO- Sug Ls Me, A Le Wy iin Li connect. each of the following is independently 25 -(CRmRak-Lyi-(CRimRn)teN(Rp)-(CRmRn)tN(Rp)-(CRinRn)i-CO-, (51) 0 -(CH2)-Li-(CH2):-N(Rp)-(CH2):-N(Rp)-(CH2)-CO-» FEAF The -CO- end of La is connected to Ls, and the other wm 5S LI of La is connected. In a further embodiment, each of L is independently -(CH»2)-NH-CO-(CH2):-N(Rp)-(CH2)-N(Rp)-(CH2)-CO-. BK -(CH2):-CO-NH-(CH2):-N(Rp)-(CH2):-N(Rp)-(CH2)-CO-, Heveze 30 -(CH2)-NH-CO-(CH2).-N(Rp)-(CH2):-N(Rp)-(CH2)-CO-, the -CO- end of EHF Ly is connected to L3, and the other end of L2 is connected to Li. In a further embodiment, Rs are each independently -CO-(CH2CH2O)m-CHs3,-(CH2):-NH-CO-(CH2CH20)m-CH3. #%-(CH2)-CO-NH-(CH2CH20)m-CH3, (itz -CO-(CH2CH20)m-CH3). In some embodiments, 2BAH E-(CRmRn)t-Lo-CH(R,y)-CO-, (9 / 20 35 “”-(CH)rLM-CHOR)-CO-, and for example -(CH)rCO-NH-CHOR)-CO- or -(CH2)+NH-CO-CH(R,)-CO-, wherein the -CO- end of Lo is connected to L3, and the other end of Lo is connected to Li. In a further embodiment, R is independently -CO-(CH2CH2O)m-CH3x, -(CH)rNH-CO-(CH2CH2O)m-CH3x, or -(CH)rCO-NH-(CHCH2O)m-CH3, preferably -(CH?)rcNH-CO-(CHCH2O)m-CH3. 16 WO 2024 / 255740 PCT / CN2024 / 098491 Preferably, L2> are independently: -(CH2)H-(CH2CH2O), -(CH2)-CO-, -(CH2z)rCO-NH-(CH2CH2O)s-(CH2)rCO-, -(CH2)rNH-CO-(CH2CH2O)s-(CH2)rCO-, -(CHz)-CO-NH-(CH2z)c(3-8 methyl aryl)-(CHz)r(OCHCH2)m-NH-CO-(CH2O)-(CH2)rCO-, -(CHz)rCO-NH-(CH)rNH-CO-(CHCH2O)m-(CH2)rNH-CO-(CH2O)-(CH2)rCO-, 5 -(CH»)-CO-NH-(CH2)-NH-CO-(CH2)-(OCH2CH2)m-NH-CO-(CH2O)-(CH2)-CO-; -(CH2)-NH-CO-(CH2):-N(R»)-(CH2):-N(Rp)-(CH2),-CO-, -(CH2):-CO-NH-(CH2)-N(R»)-(CH2):-N(Rp)-(CH2)-CO-, BY -(CH2)-CO-NH-CH(Rp)-CO- or -(CH2)ttNH-CO-CH(R,)-CO-, where the -CO- end of 1 is understood to be related to Ls EH, A Lo 55 — bing Li ERE. 10, as LwThe structural units (e.g., -NH-CO- or -CO-NH-) are arranged in the structural formula of Le from left to right. Similarly, the structural units of LN are also arranged in the structural formula of Le from left to right. In some embodiments, s are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. In a further embodiment, s are each independently 0. In some embodiments, s are each independently 3. In other embodiments, s are each independently 8. In some embodiments, t are each independently 0, 1, 2, 3, 4, 5, 6, 7, or 8. In a further embodiment, s are each independently 0, 1, 2, 3, 4, 5, or 6, preferably each independently 0, 1, 2, 3, 4, or 6. In some embodiments, m are each independently 3, 4, 5, 6, 7, or 8. In a further embodiment, m are each independently 3. In other embodiments, m is independently 8. In some embodiments, L2: is independently a connecting unit of the formula -(CHCHO)x-(CH)vCO-, wherein -CO- 2O in LER, ALi connected, and wherein x is an integer from 0 to 5, y is an integer from 1 to 5, and x+y<6; preferably, L1 is -(CH2)-CO-, -(CH)-CO-, -(CH)3-CO-, -(CH)4-CO-, -(CH2)5-CO-, -(CH2)e-CO-, -(CH2CH2O)-(CH2)-CO-, -(CH2CH2O)-(CH2)2-CO-, -(CH2CH2O)-(CH2)3-CO-, -(CH2CH2O)-(CH2)4-CO-, -(CH2CH2O)-(CH2)5-CO- , -(CH2CH20)2-(CH2)-CO--(CH2CH2O)3-(CH2)3-CO-. EVE and Le are independently -(CH?)s-CO- or -(CHCH2O)3-(CH2)-CO-. In other embodiments, L2 is independently: * -(CH2)5-CO-; 4 -(CH2CH2O)3-(CH2)2-CO-; 30 -(CH)-CO-NH-(CH2CH2O)s-(CH2)-CO-; * -(CH2)3-CO-NH-(CH2?)-(triazolyl subunit)-(CH2)-(OCHCH2)s-NH-CO-(CH2)-O-(CH2)-CO-: ¢ -(CH2)3-CO-NH-(CH2)2-NH-CO-(CH2CH2O)s-(CH2)2-NH-CO-(CH2)-O-(CH2)-CO-; ¢ -(CH2)3-CO-NH-(CH2)2-NH-CO-(CH2)2-(OCH2CH)2)s-NH-CO-(CH2)-O-(CH2)-CO-; HAN OY . 9 otto , 17 WO 2024 / 255740 PCT / CN2024 / 098491 OO HAN A OAS . O om 04, . H 9 oN 6) O fo hy Nh 8 4 O : Wherein the -CO- end of Lo is connected to Ls and the other end of Lo is connected to LI, or where position 1 is connected to Ll ve and position 2 is connected to Ls. 5 In some embodiments, L3 is independently an amino acid residue, a short peptide chain consisting of 2-6 amino acid residues, or -NH-(CH2)-CO-, where p is 0, 1, 2, 3, 4, 5S or 6. Specifically, the amino acid is a natural or non-natural amino acid, preferably selected from glycine (GI), alanine (Ala), citrulline (VaD), alanine (Phe), citrulline (Cib), lysine (Lys), and asparagine (Asm). In some embodiments, L3 is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Lys, Ala-Ala-Ala, Ala-Ala-Asn, or -NH-(CH2)-CO-. Specifically, L3 is...Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Val-Lys, Ala-Ala-Ala, Ala-Ala-Asn, or -NH-(CH2)-CO-. In some embodiments, L3 is Gly-Gly-Phe-Gly, Val-Cit, Val-Ala, Phe-Lys, Lys or -NH-(CH2)-CO-. Specifically, IL53 is Gly-Gly-Phe-Gly, Val-Ala, Phe-Lys or s. 15 In some embodiments, each IL4 is independently: OH ro Os fe) fe) | : OH A N. & 6 BS ote So INT tyre Yo «ley a covalent bond, .¢ , H , H and H 0 , wherein the 1-position is connected to L3, and the 2-position is connected to D. fe) | SS o Ne In further embodiments, each L4 is independently a covalent bond, HH , 4H or ce) OH ot any Os : OH 口 was " o , wherein the 1-position is connected to L3, and the 2-position is connected to D. , , AN 一 , , , , 20 In some embodiments, each L4 is independently OH , wherein the 1-position is connected to L3, and the 2-position is connected to D. 18 WO 2024 / 255740 PCT / CN2024 / 098491 In some embodiments, the -L1-L2-L3-L4- moiety is each independently selected from: N Ns) O O O re) WD (MC-GGFG-) ; 9 H 了 H 量 H 5 O OO \ O 5 3 O O O 4 9 cre 6 H 0 oa x 9 NH, (MC-VC-PAB-) ; Fi OH ,, OH HOT "Ss ous) ~~ OH O fe) wens NAA HK AK, SP 6 H H 9 , and 10 天、 一 人、 N Ns) O O O O 1 (MC-PEG3-GGFG) ; wherein the 1-position is connected to Ab, and the 2-position is connected to D. In some embodiments, the -L1-L2-L3-L4- moiety is each independently selected from: N N N No NON ofp Ain N H a oO (~ 8oY Ho 15 O , 19 WO 2024 / 255740 PCT / CN2024 / 098491 Oo NOAA O O = O H 9 H 9 one one one ro person all NN ne O H fe) = O the person mI , N > N NAVAS H O = H N ee \ = H _ H ot yo I N=N N N HN? ® NH person WI g 7 Oo O by O WN NO yo, S) N,, ne H 0 H H and H oO 5 NH2 , A a A N HN,, es O O N NH> , O O O H er 9 H 9 all all person NAN O = H O O O 7 so O 4H 9 O = O ook O 20 WO 2024 / 255740 PCT / CN2024 / 098491 ONS | oar TNE 0 4H 9 SS Ny ANA, N person “eo O ALN H : h O of 9 , and O O O H H O o = 4 O £0 NH 8 O many Jip, position 1 is connected to Ab, position 2 is connected to D PERE. In some embodiments, the -L1-L2-L3-L4-D moieties are each independently selected from: 23° oO NOAA IL N H 9 O oO N O O a N = \ ff 0 F 2 STI-A3" HoO _O 24° O NOAA N H 9 NO nN 9 O NH O O H3C a N F : HO": OO STI-F3" ™ 25° oO veut we \ 3 N N O oO NIC Sa NOP eo ew 0 O O N HN — — O H S HO 二 N STI-F4' H3C F 21 WO 2024 / 255740 PCT / CN2024 / 098491 26° O O y, N H 9 NaN _\ NI F HO = O ae — H still ° STI-G' 27° ? S oO 10) V N H Q NHN _ AN F HO = O pL ~ H “i 9 STI-G3' 28° Q O O NM \¥ H ff N — Ho O o 4 gy Ayre “7 NA Ay 7) Mn H H O O H3C F STI-F1002' 29° Q O OH oO H N a O N ty O see es “7 9 HaC F STI-F1001' 22 WO 2024 / 255740 PCT / CN2024 / 098491 30’ or NH O O fe) Oo H wey N ‘ou, a HO N O | a O O N OO H3C STI-F1003' F NH O O O H O O H \ 一 一 一 f N “CL N\ ye N H O. UN HO O 9 OY ce) oO N HC STI-F1004' F 32’ re) O ML O 4 O H 9 we AN 9 N “nN OO O O F STI-F1006 33’ 0 O O O H 9 4 N \ 9 O H oO N STI-F1010 HaC F 34’ O O O 4 O 4H 9 nN \ 9 H _ ft eee re eS Cee, ZA HO fe) H O : | 5 N STI-F1011 H3C F 35° oN O o WI ie) N \ 9 SS we H H — = SS Staff 一 2 : HO H fe) = H N STI-F1012 F 23 WO 2024 / 255740 PCT / CN2024 / 098491 36’ N ° -two H NN ae: N=N N N “o _ , oO for ~HN N L NAY, Cr oT : ° o aa v\~e NH, HN. OH N STI-F1013 H3C F 37° 4 O O 4 fe) H 9 we \ fe) N N “oo N person |aa q N “ Z — ud 一 O O | fe) N STI-F1014 H3C F 38° 5 i i we we -one person 一 8 12} N N N Oo” ayy (of Suny N i N O.. N as bb © ° a Ho | N STI-F1015 MeC 3 F 39° oN 8 Q “yr | ro) Na ~ re) H re} re) i fo) H N person 一 SS NN bOI {S) N,,, NOT A HO H g H HOT (a N STI-F1018 H3C NH F 40° Q O ; nv H 9 O H — / be N H N HO BA a N ,NE wort yhoo} Woy“% No SS ie] fe] N STI-F1019 HC i NH 41° ce] Nl fe) H fe) H 口 fe) N S Cybol Ty tA, N No N 0 O O OF NON LN STI-G1001 HO” \ — 42’ Q 0 fe) fe) H fH 了 H 9 H N a N N 一 SNaaa NA Ag ae HO ie) = H N O ott ok O STI-F1031 HaC F 24 WO 2024 / 2SS740 PCT / CN2024 / 098491 43° je) , f°}? 0 4 o ie) O NAAN Ay AON A NH Ao NM \_e O © AL ok H oO = 4H ae A 一 1 一 N STI-F1034 Hac F 44’ a Q 斑 a | fo \ 9 N72 g fe) fe) N ‘ey, Aa ORE ZO ~~“ HO 一 9 ot ol Noo FF N STI-F1035 H3C F 45’ 6 9 9 Q fe) ; Hye HN Ayo N_\ _ O 0 = 4 4 HO 9 Lo ; NH H.C ay STI-F1043 OF JOP, 1 position represents the site linked to Ab. In some embodiments, Ab is an antibody or antigen-binding fragment thereof that binds to a tumor cell surface antigen; for example, an anti-Her2 antibody or antigen-binding fragment thereof and / or an anti-FRa antibody or antigen-binding fragment thereof. In some embodiments, Ab is a monoclonal antibody or antigen-binding fragment thereof and / or trastuzumab or antigen-binding fragment thereof. re) Oo Hcy" er In some embodiments, when X is O, Lg and X together form 5. When Lg is a leaving group, the leaving group can be easily eliminated when reacting with an antibody or an antigen-binding fragment thereof. In some embodiments, the leaving group is a group including p-toluenesulfonyl), sulfonyloxy (for example, mesyloxy, CF3SO3-, pMethylsulfonyloxy), tertiary amine salt (e.g., Me or EN), or diazonium salt. Preferably, Lg is FE, CI, Br, or methanesulfonyl. More preferably, Lg is an acyl group. In some embodiments, the antibody-drug conjugate of formula four of the present invention is selected from: 25 WO 2024 / 2SS740 PCT / CN2024 / 098491 Fa-ADCI 0 fe) fe) O = NHHH NL Nhl N y 1 / OOF HO 20 n Fa-ADC2 oo O (S) OH OH 9 O NK NOOK a \ mAb N NO H Oo > O fe) F n Fa-ADC3 OOO nN SY fe) 4 station Ai 9 H OH OO HL NEN MA mAb N NL H oO OOF n Fa-ADC4 OOO \ AS) N a7) O an N - oH Pk Oh Ayo All from " ° N mAb N NOE H oO OOF n 26 WO 2024 / 255740 PCT / CN2024 / 098491 Fa-ADC5 o 0 fe) fe) This Z — OH H 9 OH Can_ One person " mAb N NO H (e) F (6) n Fa-ADC6 OO Mouth NA O HH Ho One 0 4 gyre 5 So i , O mAb NALA Ay SAN " 6 H 0 H H3C F n Tra-ADC7 fe) oy Nn \ ° 5 N i we we AN oO ~ HO N "o | mAb yO oh a NINN ° HC t Tra-ADC8 QO fe) OO NA 9 mA Do one person down N and N : HN S07 NN oO HO H3C F n Tra-ADC9 QO o 4 9 4 9 4 Nn \ Ye mAb ee eee Oe Se, a HO D ° o 8 N H3C F n 27 WO 2024 / 255740 PCT / CN2024 / 098491 Tra-ADC10 mAb. UN QO Wh 4 9 | VGN SS N NA No aHG H 6 = 4H N H3CF n Tra-ADC11 O / N N=N H N ; HN7 ~O mAb 二 = L To Q oo . 下 9 人 he, HN A OH N H3C | 四 Tra-ADC12 O © ‘ 4 fe H oO H N 从 \ oO N NK ie NO N 全 mAb aaa (ofS N I N TO) Ad ud -一 bb N H3C F n Tra-ADC13 fa O O Q H 9 H 人 由 从 一 or 4 N re) mAb yaAy jf N Ch. N 》 ce A HO ° N H3C , F Tra-ADC14 mAb an 4 O WI re) O fe) NM 全 2 no yr yey 1 N,, 6 N o 人 一 一 a y HO HC NH» F n 28 WO 2024 / 255740 PCT / CN2024 / 098491 Tra-ADC15 O fo) \ oO H ?9 O H / 一 N H N HO yr Ay to ott No “1 SN 口 mAb 1 N ° HaC NH; F A Tra-ADC16 mAb} NSN Yoh yw OY NN ao STON bb O 8 H ft H H o NA 7 5 HO 7 ‘5 Tra-ADC17 O fe) O 0 ao O mAb -上 并 COR H 9 HP Nu ohee ~ 全 全 一 YN 人 一 N NN AGN Zo HO H : ° © AL Aok oF 8 N H3C F n Tra-ADC18 O , fe 6 9 0 ° mAb NAN A OA, NIL AG vw N O O O AN Ho : 4H “TC = “一 fe) of, a HO N H3C F n Tra-ADC19 mAb 下 _-NN 9 Q Im de 一 fe) H Oo 4 N N Qe S RAR MT a HO 0 AL Aol; Hoo fF 4H N H3C F n 29 WO 2024 / 255740 PCT / CN2024 / 098491 Tra-ADC20 Oo Hg n°) 8 9 5 mAb NN Ne HN Amo N _\ _ fe) O = O HO 9 N Lo NH H2C Tay 3 if In Fa-ADCs,mAb represents farletuzumab (Fa); in Tra-ADCs, mAb represents trastuzumab (Tra); AnNHo# 8 Hee, HMO. 1, 2. 3. 4. 5. 6. 7S. In some embodiments, the antibody-drug conjugate of the present invention has an average DAR of any value selected from 1.0 to 10.0, preferably has an average DAR of any value selected from 2.0 to 8.0, and more preferably has an average DAR of any value selected from 6.0 to 8.0. In some embodiments, the compound of formula (I) of the present invention is selected from: 1 Cl 2 to ww \ { ‘o = N F HO”: nN N / O STI-A-4 全 F How: “Oo STI-A-6 全 3 H 4 H N fe) N 口 SU LA, 和 尼 \ | re) nN \ 人 Oo : F HO = 0 STIA HO" © STI-A2 人 5 OH O 0 o @ Sy N = ON o NA 7 / oY Io O N : Ho 以 F HO 20 NN STI-F-01 STI-E05 7 HO O 0 = N H3C 、 N 3 必 \ / O e aN / | D F HO 2 0 ° HG = _9 STLF-8 STI-E 30 WO 2024 / 255740 PCT / CN2024 / 098491 ie 10 H2N O fe) = N HN O HasC / O Hac 四 3 nN \ / O F HO = 0 F HO = 0 STI-F-03c-1 STI-F2 11 12 O O H2N 人 1 / = ON 2 4 \ 0 ON i re) N 2 nN’ \ F HO 2 0 F HO’ 20 STI-G1 STI-G03 13 国 14 本 S fe) S fe) Br aN io H2N aN / F Hom: 0 F Hom: 0 STI-G2-06¢ STI-G2 15 HO 16 HO 9 HN 9 O N = 、N AR || o Hof: © N F — F HO’ a fe) STI-D STI-F 17 a 18 a NH O NH O = N = N HaC i) re) FHO 20 F Ho"= 0 STI-F6 STIF7 19 20 (fe) and O=g_ \ PON NH O re) H3C y Ir %o HzN / N \ vn ~ F HO 20 F HO = 0 STI-F13 STI-G4 31 WO 2024 / 255740 PCT / CN2024 / 098491 21 HO 22 HO = N HN 9 fe) H3C o\ If o -AN N : H3C o-\ fo F HO’ 2 0 N - F HO ~_e STI-F12R ng Pl none P2 HO NH O - N NH O N H3C a If °o - N F HO z O " ~~ F HOY: O STI-FS STI-F8 P3 P4 HO N 9 NH O = N = N H3C oN / Oo H3C vs Io F HOw: 0 F HOw: oO STI-F14 STI-F15 PS HO o O HN H3C o\ I ~o a_N N O N \ 4 F HO 2 0 O STI-F17 STI-F18 HO Sb P7 oO == NN Ox HN a\ If Db HO F HOw 0 — STI-G5 In some embodiments, the drug linker conjugate of formula (I) of the present invention is selected from: 32 WO 2024 / 255740 PCT / CN2024 / 098491 23 0 jj 10) 7 O NOAA JL N H oO O O N ° O Or \ N Sy \ / D F STI-A3 HO _O 24 ae ra) fe) ry if H oO NAA SF NA H N N HN 0 vy att 0 = N H3C y io F HO = 0 STI-F3 25 0 N_ N H FO 9 口 fe) ae RP a a O & N aA — Te N H3C STI-F4 F 26 O 1 3 a N Q NN | SNY Oo F pL mu H ¢ O STI-G 27 ) S O O y N H 9 Nan _ . Io F HO: O ae: H Us 9 STI-G3 33 WO 2024 / 255740PCT / CN2024 / 098491 28 Q O oO N \y H 9 H 一 HE 一 O 6 oo NA a A ATF Nr N a7) O H H O H3C F STI-F1002 29 0 O NM AN_ / 5 HJ N 一 HO 一 fe) j ao 4 goo oO" i , O 0 H 6 H H3C F STI-F1001 NH O O O O H \ wey” H OT Nz HO N I Ho ni | \ oO N O H3C STI-F1003 F NH O O O H fe) O H \ 0 9 ona O O N H3C STI-F1004 F 32 Q Q NM NA] i we HL AN a ~ HO N N N N 人 ~ | comet(oh N if H N 9 H3C STI-F1006 F 34 WO 2024 / 255740 PCT / CN2024 / 098491 33 O O O j O O NA 0 N : HN SOT NEN a : HO O H 6 : N H3C STI-F1010 F 34 8 Q fe) 4 9 yO nN \ O N H _ 入 : Commee ee, Z : HO (e) O 〇 = N fe) H3C STI-F1011 F 35 O ~s N O fe) ng 9 | \ O NOAA O 4H 9 | N H O = H N H3C STI-F1012 F 36 o N 一 和 H 4 -过 Ho Ns ~ 人ae: N=N N Nw HNO oN d nto eet ; ° as ae N (一 HN —~ OH STI-F1013 NW 4 i H3C t 37 6 9 H q we Hk a nN \} 0 N N N oO ‘, Comet {ofa N 1 a6 L I > b N H3C STI-F1014 i 38 ° 4 fe) 4 人 wet cn Q Q N N N DO” one(of N I N Ch. N \ cs . 9 9 ZN HO N STLF1015 H3C F 35 WO 2024 / 255740 PCT / CN2024 / 098491 39 Q O O Wk N I 9 H 9? H N 2 S yO Oy Loo ON, AgeNN HOH 6 H H OO (YH person STI-F1018 Hic NH, F 40 Qe Naa) ° H 9 o H -le 一 Zé NA N , 一 -一 -AN and HO - 1 Nobo}Wo“person No i 8 As | O fe) > N 9 STI-F1019 H3C E NH, Al oO iN o H Oo H oO H fe) “= Ory oh ay SA, NNO , N O H H re) O O O N \ 4 F fe) STI-G1001 Ho”: SN fe) 42 Q King a oO for} nv \ 9 NN You N N NON Zo O fe) AN + H O = H N Oo of; H3C STI-F1031 F fe) 43 j 4 f° O H fe) Q fe) N N n JL No JL - O O : 一 “ O o Ji 一 co oO N 2 : HO N STI-F1034 H3C F 44 Ne N 9 O 4 _ ce) ° T Zz person Ru、 一 le 9 fe) fe) n \ 1 3 H O oo}, fe) N HC STI-F1035 F O 〇 45 。 6 | 0 o O N, NO er HN AON N\ person, 一 O o = 4 NA HO ° £0 NH H3C re) F STI-F1043 。 36 WO 2024 / 255740 PCT / CN2024 / 098491 In another aspect, the present invention provides a pharmaceutical composition, comprising the antibody-drug conjugate described herein, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug or isotope-labeled derivative thereof, or the compound of the present invention, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug or isotope-labeled derivative thereof, or the drug-linker conjugate of the present invention, or a pharmaceutically acceptable salt or ester thereof, solvate, tautomer, stereoisomer, prodrug or isotope-labeled derivative thereof, and a pharmaceutically acceptable carrier, diluent or excipient. In another aspect, the present inventionThe present invention provides an antibody-drug conjugate or its pharmaceutically acceptable salt or ester, solvent compound, tautomer, stereoisomer, prodrug, or isotope-labeled compound or its pharmaceutically acceptable salt or ester, solvent compound, tautomer, stereoisomer, prodrug, or isotope-labeled compound or its pharmaceutically acceptable salt or ester, solvent compound, tautomer, stereoisomer, prodrug, or isotope-labeled compound or its pharmaceutically acceptable linker conjugate or its pharmaceutically acceptable salt or ester, solvent compound, tautomer, stereoisomer, prodrug, or isotope-labeled compound ... stereoisomer, or isotope-labeled compound for the treatment or prevention of tumors. Prodrugs or isotope-labeled compounds, or drug linker conjugates as described in this invention, or their pharmaceutically acceptable salts or esters, complexes, tautomers, stereoisomers, prodrugs or isotope-labeled compounds. In another aspect, this invention provides antibody-drug conjugates as described in this invention, or their pharmaceutically acceptable salts or esters, complexes, tautomers, stereoisomers, prodrugs or isotope-labeled compounds, or compounds as described in this invention, or their pharmaceutically acceptable salts or esters, complexes, tautomers, stereoisomers, prodrugs or isotope-labeled compounds, or drug linker conjugates as described in this invention, or their pharmaceutically acceptable salts or esters, complexes, tautomers, stereoisomers, prodrugs or isotope-labeled compounds, in the preparation of medicaments for treating or preventing cancer.Applications. In some embodiments, the cancer is a solid tumor, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, digestive system cancers (e.g., intestinal cancer, upper gastric cancer, cardia cancer, esophageal cancer, appendiceal adenoma, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer), bladder cancer, melanoma, breast cancer, pectinary cancer, cervical cancer, endometrial cancer, prostate cancer, testicular cancer, central nervous system tumors (e.g., glioma, glioblastoma such as glioblastoma multiforme, glioma, or sarcoma), choriocarcinoma, oral epidermoid carcinoma, or hematologic malignancies, particularly leukemia (e.g., acute or chronic sporadic leukemia, and acute or chronic myeloid leukemia). Lymphomas (e.g., Hodgkin's lymphoma, non-Hodgkin's lymphoma, diffuse HEA B-cell lymphoma, and acute B-cell lymphoma, follicular lymphoma), multiple osteosarcomas, preferably, the 30 HE EON Ses. Zeiss Bk J Re BEI ASA Ts BAN a ABS BR TE Fs aS BOR ESC RDP In particular, the terminology of this application has the following meanings. The terms "an", "a", "the", and "the entity itself" as used herein should be understood to include both the singular and the plural, unless otherwise specified or clearly contradicted by the context. A hyphen (not between two letters or symbols) indicates the linking site of a substituent. For example, -NRaRe indicates that the group is linked to the rest of the molecule via a nitrogen atom, and -(CH2)p-NRaRp indicates that the group is linked to the rest of the molecule via -(CH2)p-. (37 WO 2024 / 255740 PCT / CN2024 / 098491) When the linking site of a substituent is relevant to the art...When it is obvious to a person skilled in the art (e.g., for elements, CN, OH, NH2 SB), “to Ty LHS Ais PAR Pa TRAP). RUC). FA(Bo aH, Tita. SBR. ARE cE” BBB BEN OE EE A] — Bi a ERAN Ee PAA ek 2a NY Sk A> EA) EAE BC SE 5, hydrocarbon group. Preferably, the valeryl group has 1-6 carbon atoms (Ci-6 alkyl), 1-4 carbon atoms (Ci1.4 alkyl), or 1-3 carbon atoms (Ci1a3 alkyl). Examples of alkyl groups include, but are not limited to, methyl (Mej), ethyl (Eb), propyl (Pn (FRIESE AS), TR (Bu) (including n-butyl, isobutyl, sec-butyl, and tert-butyl), pentyl (including n-butyl, isopentyl, neoalkyl, etc.), hexyl, heptyl, octyl, etc. The term "alkenyl" alone or as part of other groups refers to a straight-chain or branched alkenyl group consisting of methyl and oxygen atoms containing at least one double bond. Preferably, the alkenyl group has 2-6 carbon atoms (Ce alkenyl), 2-4 carbon atoms (C2-4 alkenyl), or 2-3 carbon atoms (C2-3 alkenyl). Examples of alkenyl groups include, but are not limited to, vinyl, terpenyl, alkenyl, isobutylenyl, etc. — RE EL SEB SC EEE]. fiat, RAL 2-6-7 SWRI (Cr-6 RAE). 2-4 RIF (Coat BE) BK2-3 PR 15 JRF (Coste). REE REN LFA BR EZR TES TASES TSE. ET UR FR, DOR ZE. EOI SEA IRE The terms “alkoxy” and “alkyl-O-” are used interchangeably to refer to an alkyl group as defined above, linked by an oxygen atom. Preferably, the alkoxy group has 1-6 carbon atoms.(Ci6 alkoxy), 1-4 carbon atoms (Ci4 alkoxy), or 1-3 carbon atoms (Ci3 alkoxy). Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy, isopropoxy), butoxy (including n-butoxy, sec-butoxy, isobutoxy, tert-butoxy, etc.), pentoxy (including n-oxy, isooxy, neooxy, etc.), hexoxy, heptoxy, octoxy, etc. The terms "alkathio" and "*alkyl-S-" are used interchangeably to indicate an alkyl group as defined above linked by a sulfur atom. Preferably, the alkathio group has 1-6 alkyl atoms (Ci6 alkathio), 1-4 alkyl atoms (Ci4 alkathio), or 1-3 alkyl atoms (Ci3 alkathio). Examples of thioyl groups include, but are not limited to, methylthio, ethylthio, propylthio (including n-propylthio and isopropylthio), butylthio (including n-butylthio, sec-butylthio, isobutylthio, tert-butylthio, etc.), and pentyl (including n-pentyl, isopentylthio, neopentyl). CE. PGT. Teme ARAB" Bed (REE TB RS IML. 2.36 46 5. OBRT4S) PA ERAT AN CE LAY alkyl. It is understood that when there is more than one halogen substituent, the halogen substituents can be the same or different, and can be located on the same or different carbon atoms. Preferably, the permanent alkyl group is a Cl-subalkyl, a Ci-4 permanent alkyl, or a Cl3 alkyl. Examples of 30” alkyl groups include, but are not limited to, fluoromethyl, chloromethyl, difluoromethyl, dioxymethyl, fluorooxymethyl, trifluoromethyl, SSP, Ca, OA, HRA, HACK, CRAZE, KRALL AW, etc. The term “hydroxyalkyl” refers to a group consisting of one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) alkyl groups.The hydroxyalkyl group is a alkyl group substituted with an alkyl group as defined herein. It is understood that when there is more than one hydroxyalkyl group, the hydroxyalkyl group may be located on the same or different carbon atoms. Preferably, the hydroxyalkyl group is a Ci-hydroxyalkyl, Ci-4-hydroxyalkyl, or Ci-hydroxyalkyl. Examples of hydroxyalkyl groups include, but are not limited to, hydroxymethyl, ethyl, propyl, hydroxybutyl, etc. The term "alkyl α" refers to an alkyl group as defined herein that is substituted with one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) alkyl groups. It is understood that when there is more than one cyano substituent, the alkyl substituents can be located on the same or different carbon atoms. Preferably, the chloroalkyl group is a Cl-6 fluoroalkyl, a Cl-4 alkyl α, or a Cl-3 cyanoalkyl. Examples of alkyl α include, but are not limited to, alkylmethyl, fluoroethyl, and alkylpropyl. The term "alkyl α" refers to an alkyl group as defined herein that is substituted with one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) nitrogen groups. It is understood that when there is more than one nitrogen substituent, the alkyl substituents can be located on the same or different carbon atoms. Preferably, the alkyl α is a Ci-6 fluoroalkyl. Aminoalkyl, C114 aminoalkyl, or C13 aminoalkyl. Examples of aminoalkyl include, but are not limited to, cycloalkylmethyl, cycloalkylethyl, cycloalkylpropyl, etc. The term "cycloalkyl" alone or as part of other groups refers to a fully or partially saturated non-aromatic monocyclic or bicyclic alkyl group consisting of carbon and oxygen atoms, preferably fully saturated. Preferably, the cycloalkyl group has 3-8 cyclic carbon atoms (Cs-s cycloalkyl) or 3-6 cyclic carbon atoms (C3-6 cycloalkyl). Examples of cycloalkyl include, but are not limited to, […].Cyclopropyl, cyclobutyl, cycloalkyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, etc. When a cycloalkyl group is replaced by one or more hydroxyl groups, it can be called a "hydroxycycloalkyl group", including C3-s hydroxycycloalkyl or C3-e hydroxycycloalkyl. The term "heterocyclic group" refers to a non-aromatic monocyclic or bicyclic group containing one or more, for example, 1, 2, 3, or 4 heteroatoms independently selected from N, O, or β, and whose remaining ring members are carbon, and is either fully saturated (i.e., heterocyclic valeryl) or partially saturated (e.g., heterocyclic valeryl containing one or more double bonds, heterocyclic valeryl containing one or more triple bonds, etc.). The carbon member of the heterocyclic group can be substituted with -CO- at any 15" angle, and the arbitrary N and S heteroatoms can be optionally oxidized (e.g., in NO, SO, SO₂) and the arbitrary N heteroatoms can be optionally quaternized (e.g., in [INR]Cl, [NR]OH). Preferably, the heterocyclic group is a 3-8 membered, 4-8 membered, 4-6 membered, or 4-5 membered heterocyclic group. A nitrogen-containing heterocyclic group refers to a heterocyclic group containing one or more nitrogen heteroatoms and with the remaining ring members being carbon, such as pyrrole, dioxopyrrole, pyrrolidine, pyridine, dioxopyridine, tetraoxopyridine, piperidine, etc. Examples of heterocyclic groups include, but are not limited to: aziridinyl, ethylene oxide, aziridine butyl, 20" oxadicyclic butyl, thiohexacyclic butyl, dioxopyrrole, pyrrolidinyl, etc. Pyrrolidone group, dioxonyl group, tetraoxonyl group, dioxonyl group, tetraoxonyl group, tetraoxonyl 1-oxide, tetraoxonyl 1,1-dioxide, dioxonyl group, dioxonyl heterocyclic alkenyl group, dioxonyl pyrazolyl group, Pyrazolyl, Zolbolinyl, Imidazolyl, Imidazolone, Dioxoimidoxone, Tetraoxoimidoxone, Dioxosazolyl, Pyrazolyl, Dioxosazolyl, Isosazolyl, Dioxosazolyl, Carboazolyl, Dioxosazolyl, Isozolyl, Dioxosazolyl, Dioxospyryl, Tetraoxyridyl, Piperidinyl, Piperidone, Dioxazinyl, Tetraoxazinyl, Hexahydroazinyl, Dihydrocaridinyl, Tetraaminocaridinyl, Hexahydrocaridinyl, Dioxospyrazinyl, PU Aube. RMSE. MGM, UMM E. DULCE. EME | = AUIBEM PU SUE SL, WBURIL, SRRBMRIL | —SURRSS. EME GES. MOSES OSE. SO - I. MOK, 1-0, ANSE. RSENS. AOR SPIE BURIED SE, AAFP EMER Cdk. AAS EICI ZR. DU SUS DUAL EEURE SS. SRIF REAL AY DG 30 "The carbon atom or heteroatom is attached to the rest of the molecule, provided that it is chemically feasible. The heterocyclic group may also optionally be coupled with other cyclic groups, which may be aromatic or non-aromatic and may optionally contain one or more cyclic groups. The term "aryl" refers to a monovalent carbon ring having one or more rings, preferably one or two rings, wherein at least one ring is aromatic, and the other rings (if present) may be aromatic or non-aromatic." The aryl group preferably has 6-10 ring carbon atoms (i.e., Celo aryl), or preferably 6 ring carbon atoms (i.e., C6 aryl or heterol). Representative examples of aryl groups include, but are not limited to, teryl, teryl, tetra-hydroteryl, and terylene. The term "heteroaryl" refers to a group having one ring carbon atom.One or more, preferably 1, 2, 3, 4, 5 or 6, independently selected from W, O or $, and any N heteroatom may be optionally quaternized (e.g. in [NR]CLT, [NR]T OH). The heteroaryl group is preferably 3-8 nucleotides, more preferably 5-8 nucleotides (e.g., monocyclic, e.g., nitrogen-containing). Representative examples of heteroaryl compounds include, but are not limited to: MEN 5 Sk, WME, WEEE, MPCIBRSE, WRIA, FERRE AIEEE | ​​ETAL | GEMS WIB E IE mime SG, MAWES, WWE EE, MERGE, MMSE, EME, RSE WR TWEE, Me RRIB[IYE FE, BURFI ZL (5 G0 7- BARR 6-F RFR ER. 5-RUARTG|R SE. 4-RURISIRER). ASSFIR We. FRASIER. RIESE. ARS EIBIARE. Me). SEERA. ARS EIRBIAR SE ISI FE, FSW. AIPM 2 MIKE, WSK. BH IKZE, OR SPIRMESE, ARIF IMSL WE 10 MASEL RUBE. MMR EMCEE. MMB SEES. MEMBER SE. IDRIMR SEMI ES IRIE SHE a WES. MCR EMCEE. MCR S EAE. MIR S ERE RE, ARIAS MEME FFM E. ME GE (The text then lists various aryl groups, including pyridinyl, pyridinopyridinyl, pyrazinopyrazinyl, and pyridinyl groups.) Heteroaryl groups can be attached to the rest of the compound via carbon atoms or heteroatoms, provided it is chemically feasible. Terminology: * "heteroaryl" * "heteroaryl subunit" * These are interchangeable, referring to divalent groups.As defined in this paper, heteroaryl groups. 15 The term "subunit" or "sub-group" indicates a divalent group derived by removing two oxygen atoms from the molecule. Ai "CN" RAR BUZE AE "OH" RARE AE "SH" ZANE ARE "NH2" 7 BE 20 ARABS -CO-" Be -C(=O)-" AB EE ARB -SO-" 22 78 BREE The term "-SO: - indicates a sulfonyl group. The term "COOH" indicates a carboxyl group. The term "NO" indicates a nitro group. 25. The term "leaving group" refers to an atom or functional group that readily leaves the molecule in a chemical reaction. Examples include, but are not limited to: halogens, such as F, Cl, or Br; sulfonyl groups, such as methyl sulfonyl or p-toluenesulfonyl; acyloxy groups, such as alkyl acyloxys (e.g., methanesulfonyloxy), trifluoromethylsulfonyloxy, or aryl acyloxys (e.g., p-methylsulfonyloxy); tertiary acyl groups (e.g., MesN or E); or heavy acyl groups. The expressions "optional," "optionally," or "optionally" indicate that the event described subsequently may or may not occur, and the expression includes both the occurrence and non-occurrence of the event. For example, "optionally substituted" includes both unsubstituted and substituted cases. "Optional substituent"* indicates that the substituent may or may not be present. When any variable appears more than once in the structural formula, it is defined independently for each occurrence. For example, the statement "optionally substituted by one or more substituents independently selected from…" means substituted by one or more independently selected substituents, which may be the same or different. Combinations of substituents and / or variables are permitted as long as a stable compound is produced. The terms "comprising" or "including?" refer to…Includes the described elements, integers, or steps, but does not exclude any other elements, integers, or steps. In this document, when the terms "comprise" or "include" are used, unless otherwise specified, and otherwise cover combinations of the described elements, integers, or steps. The term "substance of the invention" includes antibody-drug conjugates of formula four of the invention, small-molecule camptothecin derivatives of formula (ID) of the invention, drug linker conjugates of formula (D) of the invention, or their salts or esters, esters (e.g., hydrates), tautomers, stereoisomers, prodrugs, metabolites, or isotope labels (e.g., esters). In some embodiments, "substance of the invention" specifically refers to the compounds or conjugates of the embodiments, or their salts or esters, esters (e.g., hydrates), tautomers, stereoisomers, prodrugs, metabolites, or isotope labels (e.g., esters). The term "medicinalizable" refers to a substance or composition that, when administered to animals such as humans, does not produce adverse reactions, allergic reactions, or other unwanted reactions. The term "medicinalizable salt" refers to those salts that retain the biological effectiveness of the substance of the invention and are not biologically or otherwise undesirable. The substance of the invention may be in the form of a salt, preferably a pharmaceutically available salt. This includes acid addition salts and base addition salts. Acid addition salts can be formed from inorganic or organic acids. Inorganic acids include hydrochloric acid, hydrochloric acid, oxocyanate, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, peracetic acid, etc., while organic acids include formic acid, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, etc.PPR. ARR. AUR AIR. CAFU. ARERR. i DRT. URES EIR. 15 ANZA FAT ZZ I. NTE OT FORT FORT PE ZR AT A. ARR TERT. KR PR. PR. ERA. TR. PR. SRI. PLR and pyric acid, etc. Base addition salts can be formed from organic or inorganic bases, including but not limited to alkali metal salts, such as lithium, sodium or potassium salts; alkali metal salts, such as calcium or magnesium salts; organic base salts, such as ammonium salts formed by reacting with an organometallic compound containing an N group. Salts can be synthesized from parent compounds by conventional methods. 20 Pharmaceutically usable salts are preferred. However, other salts may also be useful, for example, in separation or purification steps, and can be used during preparation, and are therefore included within the scope of this application. The term "stereoisomer" as used herein refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, mixtures of non-enantiomers, and single diastereomers can be produced. Specific individual molecules can also exhibit geometric isomerism. The term "tautomer" refers to structural isomers with different energies that can interconvert at low energies. For example, proton tautomers (also called proton-transfer tautomers) include interconversions via proton transfer, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the recombination of some bonding electrons. In this application, solid lines, solid pivots, or dashed pivots may be used to depict valence bonds in the substances of the invention. Solid lines connecting to asymmetric carbon 30" atoms are intended to indicate...This includes all possible stereoisomers at the carbon atom (e.g., specific enantiomers, racemic mixtures, etc.). The real or imaginary modifiers attached to the asymmetric carbon atom are intended to indicate the presence of the indicated stereoisomer. In racemic mixtures, real and imaginary modifiers are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise specified, the substances of the present invention can exist in the form of stereoisomers, including cis and trans isomers, optical isomers (e.g., BS enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, shunting isomers, and mixtures thereof. The substances of the present invention can exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs). The present invention also includes prodrugs of the substances of the present invention. The term "prodrug" refers to a chemically modified active or inactive compound that, after being administered to an individual, is converted into the substance of the present invention through physiological processes in vivo (e.g., hydrolysis, metabolism, etc.). Therefore, in this application, the term *application* includes administering the prodrug of the substance of the present invention to treat various diseases or conditions, wherein the prodrug is converted into the substance of the present invention in vivo. Techniques for manufacturing and using prodrugs are known to those skilled in the art. The present invention also includes all pharmaceutically usable isotopic labels that are identical to the substance of the present invention, but with one or more atoms replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the substance of the present invention include, but are not limited to: isotopes of oxygen (e.g., 7H);BUI MC. BC and:CO), isotopes of fluorine (e.g., %CD), isotopes of minerals (e.g., 2 and 2253D), isotopes of ammonia (e.g., PN KN); isotopes of oxygen (e.g., 50, 70 KO); isotopes of BEY (e.g., β>P), and isotopes of sulfur (e.g., 8S). This invention also includes metabolites of the substances of this invention, i.e., substances formed in the body after administration of the substances of this invention. These can be produced by, for example, oxidation, reduction, hydrolysis, amidation, deacylation, esterification, defatting, enzymatic hydrolysis, etc. Therefore, this invention includes metabolites of the substances of this invention, including compounds obtained by methods that allow the substances of this invention to be exposed to mammals for a time sufficient to produce their metabolites. Some substances of this invention can exist in non-solventized and solvated forms, including hydrated forms. RIB compounds" The term "hydrate" refers to a complex of solvent molecules with the substances of the present invention. Examples of solvents that form hydrates include water, isobutyl alcohol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanol. The term "hydrate" refers to a complex in which the solvent molecules are water. Methods of solvation are well known in the art. The term "individual" or "patient" refers to an animal, preferably a mammal. Examples of individuals include, but are not limited to, primates (e.g., humans and non-human primates such as monkeys), horses, cattle, sheep, cats, dogs, rabbits, and animals such as mice and rats. In some embodiments, the individual is a human, including children, adolescents, or adults. The term "treatment" refers to the treatment or prevention of a particular disease, symptom, or disorder, (i) the reduction, improvement, or elimination of one or more symptoms of a particular disease, symptom, or disorder, and optionally (ti) the prevention or delay of the particular disease, symptom, or disorder described herein.The onset of one or more symptoms of a disorder or condition. In some embodiments, “treatment” refers to the improvement of at least one bodily parameter, which may not be perceptible to the patient. In other embodiments, “*treatment” refers to the regulation of a disease or condition from a physical (e.g., stabilizing perceptible symptoms) or physiological (e.g., stabilizing bodily parameters) or both. 25 The term “prevention” refers to the administration of one or more pharmaceutical substances, particularly the compounds of the present invention and / or their pharmaceutically acceptable salts, to an individual with a predisposition to the disease or condition in order to prevent the individual from developing the disease. The terms “inhibition” and “reduction” refer to the reduction or inhibition of a specific symptom or condition or disease, or a significant reduction in biological activity or baseline activity of a process. The term "effective amount" refers to the amount that effectively achieves the desired therapeutic or preventative effect at the required dose and for the required duration. It can be determined by the participating physician or veterinary practitioner and will vary depending on factors such as the compound, the disease state being treated, the severity of the disease being treated, the individual's age and relevant health conditions, the route and form of administration, and the judgment of the attending physician or veterinary practitioner. Generally, the "preventative effective amount" will be less than the "therapeutic effective amount." The term "formulation" or "pharmaceutical composition" refers to a composition suitable for administration to animals, preferably mammals (including humans), comprising at least one active ingredient and at least one inactive ingredient, such as a pharmaceutically acceptable excipient. The formulations of this invention can be any formulation applicable in the art, such as tablets, capsules, liquid formulations, etc. The term "pharmaceutically acceptable excipient" refers to a component in a pharmaceutical formulation other than the active ingredient that is non-toxic to the individual. Examples of pharmaceutically acceptable excipients include, but are not limited to, adhesives, osteolytic agents, lubricants, solvents, etc.Dispersion media, buffers, excipients, and anti-saturates are used to produce the indicated and / or desired products. It should be understood that the reaction producing the indicated and / or desired products may not necessarily be directly derived from the combination of the two initially added reagents; that is, one or more intermediates may be present in the mixture that ultimately lead to the formation of the indicated and / or desired products. The term “about” when used in conjunction with a numerical value indicates a range of approximately 0%, preferably 0%, more preferably 0%. Antibody-Drug Conjugates The term “antibody-drug conjugate” or “ADC” as used herein refers to a small molecule drug portion (Payload) and an antibody or its antigen-binding segment (Ab, responsible for targeting function). Sometimes also biologically active) substances obtained via linker coupling. 10 The coupling methods of linkers to antibodies or their antigen-binding fragments are known to those skilled in the art, including non-site-directed and site-directed coupling, such as, but not limited to, lysine coupling, hemigelatinic acid coupling, Thiomab technology, introduction of non-natural amino acid technology, and enzymatic methods. For example, for maleimide linkers, cysteine ​​residues on the antibody are used as the linking site for the payload. Eight free kinases are generated by reducing interchain dikinases on the antibody, which undergo a Michael addition reaction with maleimide to attach the payload, thereby producing an ADC product with a DAR value close to 8. 15 Antibody-drug conjugates can be characterized by the drug-antibody ratio (DAR). The term "drug-antibody ratio" as used herein is used to describe the drug-antibody ratio."5DAR" refers to the ratio of the small molecule drug moiety (D) coupled to the antibody or its antigen-binding fragment (Ab) to the total amount of the antibody or its antigen-binding fragment (Ab). The average drug-antibody ratio (average DAR) represents the molar ratio of the total drug molecules to the antibody molecules in the assay system. DAR can range from 1 to 20, but higher loadings are possible, depending on the number of binding sites on the antibody. Methods for determining DAR are known in the art, such as reversed-phase high-performance liquid chromatography (RP-HPLC), size exclusion chromatography (SEC-HPLC), mass spectrometry, and hydrochromatographic chromatography (HIC-HPLO). In some embodiments, the antibody-drug conjugate of the present invention has an average DAR selected from any of the following ranges: 1.0-20.0, 1.0-18.0, 1.0-16.0, 1.0-10.0, 2.0-14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0-8.0, or 2.0-8.0. In some embodiments, the antibody-drug conjugate of the present invention has an average DAR selected from any of the following ranges: 240.4, 440.4, 640.4, or 840.4. In some embodiments, the antibody-drug conjugate of the present invention has a DAR of 25. "The average DAR is approximately 2.0, 4.0, 6.0, 8.0, 10.0, or 12.0. In some embodiments, the average DAR value of the antibody-drug conjugate of the present invention is approximately 1.0-20.0, for example, approximately 1.0-18.0, 1.0-16.0, 2.0-14.0, 3.0-12.0, 4.0-10.0, 5.0-9.0, 6.0-8.0, 1.0-8.0, 2.0-6.0, for example, approximately 1.0, 1.1, 12, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4." 3.5. 3.6. 3.7. 3.8. 3.9. 4. 4.1. 4.2. 4.3.4.4. 4.5. 4.6, 30 4.7, 4.8. 4.9. 5.0. 5.1. 5.2. 5.3. 5.4. 5.5. 5.6. 5.7. 5.8. 5.9. 6.0. 6.1. 6.2. 6.3. 6.4, 6.5. 6.6. 6.7. 6.8. 6.9. 7.0. 7.1. 7.2. 7.3. 7.4. 7.5. 7.6. 7.7. 7.8. 7.9. 8. 8.1. 8.2. 8.3. 8.4. 8.5. 8.6. 8.7. 8.8. 8.9. 9.0. 9.1. 9.2. 9.3. 9.4. 9.5. 9.6. 9.7. 9.8. 9.9. 10.0, 12.0, and 16.0, and ranges with any two of these values ​​as endpoints. 35. The terms “complete antibody,” “full antibody,” or “full-length antibody” are used interchangeably herein and refer to antibody molecules that have the molecular structure of natural immunoglobulins. In the case of a conventional tetrachain IgG antibody, the full-length antibody comprises two heavy chains (T) and two light chains (L) linked together by disulfide bonds. In the case of a heavy chain antibody that has only heavy chains and lacks light chains, the full-length antibody comprises two heavy chains (HT) linked together by disulfide bonds. The term "antibody fragment" includes a portion of the complete antibody. Preferably, the antibody fragment is an antigen-binding fragment. The term "antigen-binding fragment" refers to a molecule that is different from the complete antibody, which contains a portion of the complete antibody and binds the antigen bound by the complete antibody. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; dAb (domain 5 antibody); linear antibodies; single-chain antibodies (e.g., scFv); single-domain antibodies such as VHH; bivalent antibodies or fragments thereof; or camelid antibodies. The term "antigen" refers to a molecule that elicits an immune response. This immune response may involve antibody production or activation of specific immune cells.Or both. Technicians will understand that any macromolecule, including virtually all proteins or peptides, can be used as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. In some implementations, the antigen is a tumor cell surface antigen, such as Her2 BX FRA. The term "complementary region" or "CDR region" or "CDR" is a region in the antibody variable domain that is sequence-hypervariant and forms a structurally defined loop ("hypervariant loop") and / or contains antigen contact residues (antigen contact sites). The CDR is primarily responsible for binding to the antigen epitope. The CDRs of the heavy and light chains are usually referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus. CDRs located within the antibody heavy chain variable domain are referred to as HCDR1, HCDR2, and HCDR3, while CDRs located within the antibody light chain variable domain are referred to as LCDR1, LCDR2, and LCDR3. Within a given amino acid sequence of the light chain variable region or heavy chain variable region, the precise amino acid sequence boundaries of each CDR can be determined using many well-known antibody CDRs. The assignment system is determined by any one or a combination thereof, including, for example: Chothia (Chothia et al., 1989, Nature 342: 877-883; Al-Lazikani et al., “Standard conformations for the canonical structures of immunoglobulins”, Journal of Molecular Biology, 273, 927-948, 1997), based on the three-dimensional structure of the antibody and the topology of the CDR ring; and Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, @ 4 ftz, US Department of Health and Human Services), based on the variability of the antibody sequence.Services, National Institutes of Health (1987)), AbM (University of Bath), Contact (University College London), Fx ImMunoGeneTics database (IMGT) (on imgtcines.f on the World Wide Web), and the North CDR definition based on affinity propagation clustering using a large number of antibody structures (North et al., “A New Clustering of Antibody CDR Loop 25 Con#%zions”, Journal of Molecular Biology, 406, 228-256, 2011). The term “antibody in TITgG form” refers to the IgG form to which the heavy chain constant region of the antibody belongs. For example, an antibody in IgG4 form means that its heavy chain constant region is derived from IgG4, or an antibody in IgG1 form means that its heavy chain constant region is derived from IgG1. The antibody or its antigen-binding segment applicable to this application can bind any antigen suitable for the antibody, preferably a tumor cell surface antigen, such as Her2 or FRa. 30. In some embodiments, the antibody or antigen-binding fragment thereof suitable for this application is an antibody or antigen-binding fragment thereof that binds to Her2, particularly human Her2. Any Her2 antibody or antigen-binding fragment thereof known to those skilled in the art is applicable to this invention, including but not limited to disclosed antibodies or commercially available antibodies, such as trastuzumab. In some embodiments, the antibody or antigen-binding fragment thereof suitable for this application is an antibody or antigen-binding fragment thereof that binds to FRue, particularly human FRe. Any FRe antibody or antigen-binding fragment thereof known to those skilled in the art is applicable to 35. This invention, including but not limited to disclosed antibodies or commercially available antibodies.The obtained antibody, for example, faucitizumab. In some embodiments, the antibody suitable for this application is an antibody in the form of IgG, IgG2, IgG3, or IgG4. Preferably, the antibody suitable for the ADC of this application is an antibody in the form of IgG1 or its antigen-binding fragment. 44 WO 2024 / 255740 PCT / CN2024 / 098491 In some embodiments, the antibody suitable for this application is a monoclonal antibody. In some embodiments, the antibody suitable for this invention is humanized. In some embodiments, the antibody suitable for this invention is a human antibody. In some embodiments, the antibody suitable for this invention is a chimeric antibody. In one embodiment, the antibody suitable for this application is a full-length antibody. 5. In one embodiment, the antigen-binding fragment of the antibody suitable for this application is selected from the following antibody fragments: Fab, Fab', Fab'-SH, Fv, single-chain antibodies (e.g., scFvV), (Fab')2, single-domain antibodies such as VHH, dAb (domain antibody), linear antibodies, haptens, or bispecific, trispecific, tetraspecific, and other multispecific antibodies. In some embodiments, the antigen-binding fragment of the ADC suitable for this application includes, but is not limited to, VHH, Fv molecules, scFv molecules, Fab molecules, and Fab')2 molecules. 10. In one embodiment of the present invention, the antibody or its antigen-binding fragment suitable for this application is trastuzumab or fastatin or its antigen-binding fragment. In one embodiment, the antibody or its antigen-binding fragment comprises one or more CDRs (preferably 3 CDRs, i.e., HCDR1, HCDR2, and HCDR3; or LCDR1, LCDR2, and LCDR3, more preferably 6 CDRs, i.e.,HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3), or VH and / or YL containing trastuzumab or fortuzumab or their antigen-binding fragments, or containing the heavy chain and / or light chain of said antibody. In some embodiments, the anti-Her2 antibody or its antigen-binding fragment of the present invention contains heavy chain variable regions CDR1, CDR2, and CDR3 from trastuzumab, and light chain variable regions CDR1, CDR2, and CDR3. In some embodiments, the anti-Her2 antibody or its antigen-binding fragment of the present invention contains heavy chain variable regions and light chain variable regions from trastuzumab. In some embodiments, the anti-Her2 antibody or its antigen-binding fragment of the present invention contains heavy chain 20 RUA HE from trastuzumab. In some embodiments, the anti-FRe antibody or its antigen-binding fragment of the present invention contains heavy chain variable regions CDR1, CDR2, and CDR3 from fortuzumab. And light chain variable regions CDR1, CDR2, and CDR3. In some embodiments, the anti-FRea antibody or its antigen-binding fragment of the present invention comprises heavy chain variable regions and light chain variable regions derived from fatatumozymec. In some embodiments, the anti-Her2 antibody or its antigen-binding fragment of the present invention comprises heavy chain and light chain variable regions derived from fatatumozymec. Linker The term “linker” as used herein is a linking portion for linking a small molecule cytotoxic drug (payload) to an antibody or its antigen-binding fragment (Ab), and may be represented in this application as a -LiL2-L3-L4- portion. Linkers typically include cleavable linkers and non-cleavable linkers. Cleavable linkers utilize the difference in conditions between the bloodstream and tumor cells (e.g., pH, protease hydrolysis, or glutathione concentration) or the specific action of the antibody in deep enzymocytes.(For example, lysosomal proteolytic enzymes) break down, releasing the payload and exerting the drug's activity. Unbreakable linkers rely on antibody degradation within lysosomes to release the drug and exert its activity. Linkers usable in this invention include both breakable and unbreakable linkers, such as acid-sensitive linkers (e.g., β-35' linkers), glutathione-sensitive linkers (e.g., disulfide linkers), enzymatically cleavable linkers (e.g., short peptide linkers, β-glucanyl nucleotide linkers), acetylamine linkers, thioether linkers, etc. In particular, linkers usable in this application include, but are not limited to, those given above, especially those used in the examples. The term "connector unit" is a component (LD) of the linker (-Li-L2-L3-L4- part), whose function is to connect the linker or a linker that is part of a drug linker conjugate to an antibody or its antigen-binding fragment. Examples include, but are not limited to, O yn? NN vw CLAN oem aoe . oi )=* ot ya ot nN I, | Fim, where position 1 is connected to Ab, and position 2 is connected to . The term "linker unit" refers to the component (Lz) of the linker (-Li-Lz-L3-L4- portion), which functions to link the linker unit (LU) to the Ls portion, which consists of amino acid residues, short peptide chains, or -NH-(CH2z)p-CO-. In this application, the component Ls of the linker is an amino acid residue, a short peptide chain consisting of 2-10 (preferably 2-6) amino acid residues, or -NH-(CH2)p-CO-, such as -NH-(CH2)-CO-. The amino acid can be either natural or non-natural, such as glycine (GI), alanine (Alt), phenylalanine (VaD), phenylalanine (Phej), citrulline (Citrine).Citrulline (CiD), lysine (Lys) and asparagine (Asn). L3 may be directly linked to the small molecule drug moiety, or linked to the small molecule drug moiety via a spacer unit. The term "spacer unit" is a constituent part (L) of the linker (-Li-L-Ls-L4- moiety). When present, it functions to link the linker to the small molecule drug moiety (D), or may provide an additional structural moiety to facilitate release of the small molecule drug moiety from the ADC. Examples thereof include, but are not limited to, the 2-position of is connected to D. As used herein, the terms "small molecule drug moiety", "small molecule cytotoxic drug moiety" and "Payload" are used interchangeably to refer to the component responsible for killing tumor cells in an antibody-drug conjugate or drug-linker conjugate. After administration, the ADC undergoes cleavage in the intercellular space or within target cells, releasing the small molecule drug or a derivative thereof, which then exerts its biological activity. To avoid ambiguity, it should be pointed out that "drug" does not only refer to "pharmaceutical products" approved by medical regulatory authorities, but also includes any compound with potential biological activity in clinical practice, research and development, or academic research. In the present invention, the small molecule drug moiety is derived from a camptothecin derivative of formula (I), particularly from the camptothecin derivatives in the examples herein. It is a moiety obtained by losing one atom (e.g., an oxygen atom) or an atomic group from the camptothecin derivative of formula (I). For example, the small molecule drug moiety is a moiety obtained by losing one atom attached to a nitrogen or oxygen atom from Ri or Ri, or by losing one oxygen atom from the hydroxyl group shown in formula (I). The hydroxyl group shown in formula (I) isThe hydroxyl group indicated by the arrow in the following structure: R, O Ww “Lo Ra HO” Oo (Il) A 46 WO 2024 / 255740 PCT / CN2024 / 098491 The small molecule drug moiety of the present invention can be linked to the linker by any chemically permissible bonding method, including but not limited to amide bonds, aminomethyl ether bonds and hydrogen tetraester bonds. LICH BIR 5 The substances of the present invention include antibody-drug conjugates of formula (ID), camptothecin derivatives of formula (ID), and drug linker conjugates of formula (ID), or pharmaceutically acceptable salts or esters, complexes, tautomers, stereoisomers, or isotope-labeled forms thereof, which have high cytotoxic activity and can be used to treat or prevent tumors such as cancer, including but not limited to solid tumors, particularly lung cancer (e.g., small cell lung cancer, non-small cell lung cancer), liver cancer, and digestive system cancers (e.g., colorectal cancer, upper gastric cancer, cardia cancer, esophageal cancer, etc.). (The remaining text appears to be a list of names and publications, which are not translated as they are not part of the main text.) Glioblastoma, glioma, or sarcoma; perichoriocarcinoma; oral epidermoid carcinoma; or hematologic malignancies, particularly leukemia (e.g., acute or chronic myeloid leukemia, acute or chronic granulocytic leukemia), lymphoma (e.g., Hodgkin's lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, acute B-cell lymphoma, follicular lymphoma), and multiple 15" osteosarcomas. Preferably, the substances of the present invention can be used for the treatment and / or prevention of pear cancer, colon cancer, lung adenocarcinoma, or oral epidermoid carcinoma. [The text then abruptly shifts to a discussion of camptothecin derivatives, which seems unrelated and likely refers to a different topic.]Compared to biological agents such as SN-38 and DXD, the substances of this invention exhibit stronger antitumor activity and / or fewer side effects. The substances of this invention are well-tolerated, effectively killing tumor cells without causing significant behavioral abnormalities or weight fluctuations (such as weight loss). The substances of this invention have good solubility and physicochemical stability, and can be easily formulated into drug formulations suitable for administration. The ADCs of this invention have a suitable drug-to-antibody ratio, and the distribution of this ratio is uniform. The ADCs of this invention are stable in blood circulation, reducing drug molecule loss in non-tissue regions and mitigating detoxification. Moreover, the ADCs of the present invention exhibit good tumor tissue targeting, accumulate in the tumor microenvironment, increase the concentration ratio of the active drug molecules within the tumor and in the blood, reduce the mechanism-related toxicity of the conjugates, and have a good therapeutic index. The small molecule drugs released from the ADCs of the present invention can also exert a bystander effect, further killing tumor cells surrounding the target tumor cells that do not express or express low levels of antigens. The ADCs of the present invention have better therapeutic or preventive efficacy and / or lower side effects than corresponding small molecule camptothecin derivatives, and have a wider therapeutic window. The substances of the present invention can be administered in the form of pharmaceutical compositions via any applicable route, such as, but not limited to, oral (e.g., in the form of tablets, gels, liquid formulations such as solutions, emulsions, or wettable powders), inhalation (e.g., in the form of sprays), rectal (e.g., in the form of suppositories), or parenteral (e.g., in the form of injections, such as intravenous, intramuscular, subcutaneous, intraperitoneal, intracranial, etc.). Parenteral administration is particularly preferred, for example, intravenous injection. The substances of the present invention are formulated into pharmaceuticals.The technology for the composition is well known in the art. For example, the substances of the present invention can be processed into pharmaceutical compositions, such as tablets, capsules, liquid formulations (e.g., injections, infusions, syrups, emulsions, suspensions, etc.), powders for injection, dispersions, sprays, suppositories, solvents, stabilizers, humectants, emulsifiers, preservatives, sweeteners, colorants, flavoring agents, salts for altering osmotic pressure, powders, masking agents, or antioxidants, etc. Dosage can be varied over a wide range and, of course, must be adjusted according to the individual needs of each specific case. The appropriate dosage of the herb can be determined by the attending physician based on the type and severity of the disease being treated, the individual's health status and medical history, the specific compound and route of administration, and any concomitant medications. The daily dose for a 70 kg adult is typically about 0.01 mg to about 1000 mg of the substance of the present invention or a corresponding amount of pharmaceutically acceptable salt or ester. The dosage of the substance of the present invention may exceed this range as needed. The daily dose may be administered as a single dose or in divided doses. The substance of the present invention can be used alone or in combination with one or more other active agents or therapies, which may have the same or different pharmacological effects as the substance of the present invention. The substance of the present invention can be administered simultaneously, separately, or sequentially with other concomitant active agents via the same or different routes of administration. They can be contained in the same pharmaceutical composition (fixed composition) or in separate forms, such as in a cassette.Products. They may be formulated and / or supplied by the same or different manufacturers. When the substances of the present invention are administered in combination with other active agents, the dosage of the combined active agents will naturally vary depending on factors such as the co-administered drugs, the condition to be treated, the individual's general health condition, and the judgment of a physician or veterinarian. To help patient compliance, the kits of this application may include instructions for use. General Preparation Methods The substances of the present invention may be prepared by a variety of methods, including the methods described in the following processes, the methods given in the examples, or similar methods. For each reaction step, appropriate reaction conditions are known to those skilled in the art or can be readily determined. Raw materials are generally commercially available or can be readily prepared using methods known in the art or methods described herein. Variables in the general formulas have the meanings defined herein unless otherwise stated. In the preparation of the compounds of the present invention, protection of groups (e.g., amino protecting groups, hydroxyl protecting groups) may be required, which can be readily determined by those skilled in the art. For a general description of the protecting group and its uses, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991. 25 UR a Bh Ee NSC RE PAY JOS) DT) (48 DAA as BANE AT oo TS NL, Pa Techniques include, but are not limited to, filtration, steaming, crystallization, chromatography, etc. The materials can be characterized by conventional methods, including physical constants and spectroscopic data. For illustrative purposes only, the following procedures provide an exemplary route for synthesizing the substances of the present invention. Those skilled in the art will understand that other synthetic routes are also available, and the compounds prepared by the methods described below...The product can be further modified based on the content of this application and using conventional compounds well known to those skilled in the art. The process for the synthesis of camptothecin derivatives is as follows: [Formula omitted - likely a typo]. The variables are as defined herein. The reaction can be carried out via Friedl der azazoline synthesis, for example, under the catalysis of an acid, such as p-toluene, glacial acetic acid, or another Lewis acid, to obtain the reaction product. The resulting product can then be further modified, for example by substitution, condensation (formation of amide bonds), Buchwald-Hartwig coupling, or amino deprotection, to produce other camptothecin derivatives. The reaction conditions are readily determined by those skilled in the art of organic synthesis. The synthesis of drug linker conjugates is within the capabilities of those skilled in the art of organic synthesis. For example, drug linker conjugates can be synthesized via Williamson ether synthesis, condensation reactions, etc. The methods for linking antibody-drug conjugates are known in the field of ADCs, such as lysine conjugation, cysteine ​​conjugation, Thiomab technology, non-natural amino acid technology, enzyme catalysis technology, etc. In particular, the antibody can be reduced first, and then the antibody can be conjugated to the drug conjugate, optionally followed by purification of the resulting antibody-drug conjugate. Figures 1a to 1d show the killing efficacy of the ADCs of the present invention against Capan-1 cells; RLU represents relative light units (Example B). Figures 2a to 2d4 show the killing efficacy of the ADCs of the present invention against NCIN87 cells: RLU represents relative light units (Example B). Figures 3a to 3e show the ADCs of the present invention.The bystander effect is observed (Example 0). Figure 4 shows the inhibition of human follicular carcinoma SKOV-3 tumor growth by the ADCs of the present invention in a mouse model (Example D.2). Figure 5 shows that the ADCs of the present invention did not cause weight loss in the SKOV-3 immunodeficient mouse model (Example D.2). Figures 6a and 6b show the tumor growth curves (6a) and weight change rate curves (6b) after administration of the ADCs of the present invention in JIMT-1 and tumor-bearing mouse models, respectively (Example D.3). Figures 7a and 7b show the tumor growth curves (7a) and weight change rate curves (7b) after administration of the ADCSs of the present invention in the Capan-1 tumor-bearing mouse model, respectively (Example D.3). Figures 8a and 8b show the tumor growth curves (8a) and body weight change rate curves (8b) after administration of the ADCs of the present invention in the NCI-N87 tumor-bearing mouse model, respectively (Example D.3). Figures 9a and 9b show the tumor growth curves (9a) and body weight change rate curves (9b) after administration of the ADCs of the present invention in the JIMT-1 tumor-bearing mouse model, respectively (Example D.4). Figures 10a and 10b show the tumor growth curves (10a) and body weight change rate curves (10b) after administration of the ADCs of the present invention in the Capan-1 tumor-bearing mouse model, respectively (Example D.4). Figures 11a and 11b show the tumor growth curves (11a) and body weight change rate curves (11b) after administration of the ADCs of the present invention in the NCLN87 tumor-bearing mouse model, respectively (Example D.4). Figures 12a and 12b show the tumor growth curves (11a) and body weight change rate curves (11b) after administration of the ADCs of the present invention in the Calu-3 tumor-bearing mouse model, respectively. The tumor growth curve (12a) and the weight change rate curve (12b) after the tumor growth curve (Example D.9) are shown. Figures 13a and 13b show the tumor growth curve in OE-19 tumor-bearing mice.The tumor growth curve 35 (13a) and body weight change rate curve (13b) after administration of the ADCs of the present invention in the model (Example D.5). Figures 14a and 14p show the tumor growth curve (1440) and body weight change rate curve (14b) after administration of the ADCs of the present invention in the Capan-1 tumor-bearing mouse model (Example D.6), respectively. 49 WO 2024 / 255740 PCT / CN2024 / 098491 Examples The following examples are provided to further illustrate the present invention. It should be understood that they are only for better understanding of the present invention and are not intended to limit the scope of the present invention in any way. In this application, when the chemical name and structural formula are different, the structural formula shall prevail, unless the chemical name rather than the structural formula can be inferred from the context to be correct. For simplicity, not all oxygen atoms are explicitly marked in some compound structural formulas given in this application. When there is a vacancy in the compound, it indicates the presence of an unmarked oxygen atom. Unless otherwise specified, the experimental materials and reagents used in the examples are commercially available or can be readily prepared using methods known to those skilled in the art. 10 Abbreviations The abbreviations used herein (e.g., for chemical groups and compounds) generally have meanings known in the art, unless otherwise indicated. For example, the following abbreviations are used herein: Acetyl (Ac), BCl, tert-butyloxybyl (Boc), chloroform (CDCIJ), NJN-dicyclohexyldiimide (DCCJ), 1,2-dioxane (DCE), dioxomethane (DCM), NJN-diisopropylethylamine (DIPEADPEA), 4-dimethylaminopyridine (DMAP), NN-dimethylformamide (DMFJ), dimethyl phthalide (DMSO), 1,2-dimethyl phthalide (DMSO), 1,2-dimethyl phthalide (DMSO), 1,2-dioxane (DCCJ), 1,2-dioxane (DCE), dimethyl phthalide (DMSO), 1,2-dioxane (DCCJ ...[DMSO-d], Ethyl acetate (EA), 6-(maleimide)hexanoic acid succinimide ester (EMCS), Triethyl ester (EN), 9-ethylmethoxymethyl ester (Fmoc), Glycine (Gly or GO), 2-(7-azodecanotriazole)-NINUNN-tetramethylurea hexafluorophosphate (HATU), Lysine (Lys or nitric acid, mol / L GO, m- and oxyperoxycarboxylic acid (mCPBA), Acetonitrile (MeCN), N-hydroxysuccinimide (CNHS), Trifluoromethanesulfonate (OT and, tri(di(20WAKSE EN TAM) — 440) [Pd(dba)j], 1,1-bis(di-tartrate) diammonium ferric chloride [Pd(dppbCl], Petroleum ether (PE), Pentafluorophenol FP-OHJ, Coalanine (Phe Or J, p-methylthiazoline pyridine salt (PPTS), preparative thin-layer chromatography (Prepb-TLCJ), pyridine (Py), trifluoroacetic acid (TFA), trifluoromethanesulfonic acid ester (TPO), thin-layer chromatography (TLCJ), p-methyl-co-acyl (Ts), p-methylthiazoline sulfonic acid (TsOHJ), 4,5-bis(didecyl)-9,9-dimethyloxazine (Xantphos) and sodium triacetyloxysine (STAB). 25 Basic information on detection and preparation: Liquid chromatography-mass spectrometry (LCMIS) instrument model: Agilent 1260II G6125B; test method: electrospray ionization source positive / negative ion mode (ESD); 30 Column model: YMC-TRIART C18 (4.6x50mmx5Shm); mobile phase: A (10Mm NH4HCO3) — B (MeCN): 0-1.5 min (5%B-100%B), 1.5-2.5 min (100%B-100%B), 2.5-2.6 min (100%B-5%B), 2.6-3.75 min (5%B-5%B); Flow rate: 2.0 mL / min; FEY: 40°C. LCMS data were generated using the following conditions: SUA RRA PCL JRA IR A Br. 35Nuclear magnetic resonance spectroscopy (LUH NMR) was performed using a Bruker AVANCE NEO 400 NMR spectroscopy system. The MAA was YARN DMSO-ds (+D20)8K CDCHb, with tetramethylsilane (TMS) as the internal standard. The signal multiplicity was indicated by the following abbreviations: s, HUE; d, — EI; t, =H; gq, PUI; dd, M (doublet), dt (doubletuplet), tt (tripletuplet), td (tripletuplet), ddd (double doubletuplet), br (broad peak); heptet, m (multiplex). All observed coupling constants J are reported in Hertz. Exchangeable protons are not always observed, including but not limited to reactive oxygen species such as hydroxyl oxygen, hydroxyl oxygen, and nitrogen-based hydrogen. 5. Medium- and High-Pressure Reverse Preparation Instrument: SHIMADZU LC-20AP; Column: SHIMADZU C18 (50x250mmx10hm); Acidic mobile phase: 0.05% TFA aqueous solution - MeCN; Alkaline mobile phase: 0.1% NH4HCO3 aqueous solution - MeCN; 10" Neutral mobile phase: H2O - MeCN; Flow rate: 15 mL / min; Detection wavelength: 214nm, 254nm. Medium- and Low-Pressure Reverse Preparation Instrument: ISOLERA PRIME 15; Column: Spherical C18 (40-750 pm, 100 A); Alkaline mobile phase: 0.1% NH4HCO3 aqueous solution - MeCN; Acidic mobile phase: 0.05% TFA aqueous solution - MeCN; Flow rate: 25 mL / min; Detection wavelength: 214nm, 254nm. Example 1, Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-1L-(methyl)-112-dichloro-14H-pyrrolidine[3144':6,71 pyrazino-20 [1,2-b] SHK-3,14(4H)-— Be] CSTI-A-4)Step 1: Under nitrogen and ice bath conditions, add 160 mL of DCE and 1M boron trioxide DCM solution (32.0 mL, 32.0 mmol), then add compound 1a (5.0 g, 40.0 mmol, 0°C) and stir for 10 minutes. Continue stirring in an ice bath with PINAR ZR (5.4 mL, 47.0 mmol, 0°C) and aluminum trichloride (7.0 g, 52.0 mmol), then slowly raise the reaction system to room temperature (25°C). Stir at room temperature for 10 minutes, then reflux and stir for 39 hours. After the reaction is complete, add the system to 80 mL of ice water, then add 40 mL of 4M HCl and stir at room temperature for 30 minutes. Add 80 mL of dioxane, and concentrate the organic phase with anhydrous sodium aluminate under reduced pressure. The residue is then prepared by reverse mixing under medium and high pressure (neutral mobile phase). After lyophilization, 1.0 g of compound 1b was obtained as a yellow solid powder, with a yield of 12.4%. LCMS (254 nm, purity 97.2%, Rt = 1.789 min; MS calculated value: 201.0; MS measured value: 202.0 [M+H]*. 'H NMR (400 MHz, DMSO-d6) 5 7.68 (d J = 8.7 Hz, 1H), 7.30 30 (s, 2H), 6.53 (d, J = 12.5 Hz, 1H), 4.97 (s, 2H), 2.08 (s, J = 8.6 Hz, 3H). Step 2: Compound 1b (500 mg, 2.48 mmoD) and compound 1c (490 mg, 1.86 mmoD) were dissolved in 30 mL of anhydrous methyl methacrylate. Under nitrogen protection, refluxed for 30 minutes using a water separator. Added p-methylthiazolic acid (48 mg, 0.25 mmol), ARSED 51 WO 2024 / 255740 PCT / CN2024 / 098491 PE 3.5" IN. TLC (DCM:MeOH = 30:1, product Rf = 0.4). The reaction was monitored by 40 LCMS. After completion, the reaction system was cooled to room temperature and analyzed.Filter, wash with acetone, vacuum-dry the filter cake at room temperature to obtain 719 mg of compound STI-A-4 as a yellow solid, with a yield of 90.2%. LCMS (254 nm, purity 96.8%, Rt = 1.711 min; MS calculated value: 428.1; MS found value: 429.2 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (d, J = 8.1 Hz, 1H), 7.94 (d, J = 10.7 Hz, 1H), 7.32 (s, 1H), 5.44 (d, J= 5.1 Hz, 4H), 5.35 (s, 2H), 2.54 (s, 3H), 1.92 — 1.78 (m, 2H), 0.88 (t, J = 7.3 Hz, 3H). Example 2: Synthesis of (S,E)-4-ethyl-4-hydroxy-8-chloro-9-methyl-11-((neopentylimino)methyl)-1,12-dihydro-14H-pyrido[3',4':6,7]indolo[1,2-b]quinoxaline-3,14(4H)-dione (STI-A-6) 1) TSOAg, DMSO 2) 90°C STI-A-4, 2a → STI-A-6 Compound STI-A-4 obtained in Example 1 (500 mg, 1.17 mmol) and Cs₂CO₃ (390 mg, 1.40 mmol) were dissolved in 30 mL of DMSO, and stirred overnight at room temperature under nitrogen protection; then heated and stirred at 90°C for 3 hours. After LCMS monitored the completion of the reaction, the reaction mixture was cooled to room temperature, p-toluenesulfonic acid (21 mg, 0.12 mmol) and compound 2a (234 mg, 2.69 mmol) were added, and stirred at room temperature for 3.5 hours. After LCMS detected the completion of the reaction, the title compound (STI-A-6) was obtained by medium-low pressure reverse-phase preparation (alkaline mobile phase) and lyophilization, 152 mg, as a yellow solid, with a yield of 27.3%. LCMS (254 nm, purity 94.8%, Rt = 2.234 min; MS calculated value:477.2; MS Found: 478.1 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 9.29 (s, 1H), 8.56 (d, J=8.1 Hz, 1H), 7.78 (d, J = 10.6 Hz, 1H), 7.22 (s, 1H), 6.45 (s, 1H), 5.34 (s, 2H), 5.17 (s, 2H), 3.56 (s, 2H), 2.42 (s, 3H), 1.87 — 1.76 (m, 2H), 1.00 (s, 9H), 0.83 (t, J= 7.4 Hz, 3H). Example 3: Synthesis of (S)-4-((1-(tert-butylpropyl)-4H-[3',4':6,7]pyrido[1,2-b]indole-3,14(2H)-dione) (STI-A) N o Ho, Pd / C nN 0 — — — = N — N Ha State D F HO x O STI-A-6 STI-A After compound STI-A-6 obtained in Example 2 (152 mg, 0.32 mmol) and 10% Pd / C (51 mg, 0.05 mmol) were reacted in 10 mL of methanol at room temperature under 1 atmosphere of hydrogen, the reaction was monitored by LC-MS till completion. The reaction system was filtered, and the filtrate was subjected to medium-low pressure reverse phase preparation (alkaline mobile phase), and lyophilized to obtain 37 mg of the title compound (STI-A), which was a yellow solid, with a yield of 24.2%. LCMS (254 nm, purity 99.0%, Rt = 2.147 min; MS calculated value: 479.2; MS found: 480.3 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (d, J= 8.5 Hz, 1H), 7.85 (d, J= 10.8 Hz, 1H), 7.31 (s, 1H), 6.51 (s, 1H), 5.43 (s, 2H), 5.38 (s, 2H), 4.30 (s, 2H), 2.49 (s, 3H), 2.43 (s, 2H), 1.93 — 1.81 (m,2H), 0.88 (s, 12H). 5 Example 4: Synthesis of (S)-4-Z,3k-4- FRE 8-FR-9- FE 1-H EE A AE) FB) -1,12-—A-1 4 SF [3',4':6,7] 95] ESF [1,2-b] EWK-3,14(4H)-— A CSTI-A2) == ON rk To MeOH aa on Ww VL 3) N : F Hore Ny (Nie , TSOH F Hot: Pb F Ho"=_ © AL 人 4a 4b STLA2 Step 1: Compound STLA-4 (150 mg, 0.35 mmol) obtained in Example 1 and silver p-toluenesulfonate (117 mg, 0.42 mmol) were dissolved in 10 mL of DMSO, and the mixture was stirred overnight at room temperature under nitrogen protection; then the mixture was heated and stirred at 90°C for 3 hours. After LCMS monitoring indicated the reaction was completed, the mixture was cooled to room temperature, p-toluenesulfonic acid (3 mg, 0.02 mmol) and compound 4a (73 mg, 0.74 mmol) were added, and the mixture was stirred at room temperature for 1 hour. After LCMS detection indicated the reaction was completed, purification was performed by medium-low pressure reverse-phase preparation (alkaline mobile phase), and lyophilization gave 16 mg of compound 4b as a yellow solid, with a yield of 10.4%. LCMS (254nm) purity 95.5%, Rt = 2.084 min; MS calculated value: 489.2; MS found value: 490.2 [M+H]+. ¹H NMR (400 MHz, DMSO-d6) δ 9.45 (s, 1H), 8.73 (d, J= 8.1 Hz, 1H), 7.94 (d, J= 10.9 Hz, 1H), 7.32 (s, 1H), 6.52 (s, 1H), 5.42 (s, 2H), 5.33 (s, 2H), 2.52 (s, 3H), 1.89 — 1.78 (m, 4H), 1.72 — 1.62 (m, 2H), 1.63 — 1.48 (m, 3H), 1.49 — 1.37 (m, 4H), 0.88 (t, J= 7.3 Hz, 3H). Step 2: Compound 4b (10 mg, 0.02 mmol) and 10% Pd / C (3mg, 0.003 mmOD was suspended in 3 mL methanol and stirred at room temperature for 6 hours under nitrogen (1 tm). After the reaction was completed, the system was filtered, and the filtrate contained 3 mg of the title compound (STLA2) prepared by low-pressure reverse radiation (MIE A), UEFA SI, and LCMS (254 20 nm). The purity was 95.7%, Rt = 1.959 min; MS calculated value: 491.2; MS measured value: 492.3 [M+H]-. !IH NMR (400 MHz, DMSO-de) 5 8.32 (d, J = 8.5 Hz, 1H), 7.88 (d, J = 10.7 Hz, 1H), 7.57 (s, 1H), 5.32 (s, 2H), 4.80 — 4.61 (m, 2H), 4.35 (s, 2H), 4.19 (d, J= 13.5 Hz, 1H), 2.65 (d, J= 7.5 Hz, 2H), 2.47 (s, 3H), 2.09 — 2.01 (m, 2H), 1.72 (d, J= 14.2 Hz, 3H), 1.52 (d, J= 22.0 Hz, 6H), 0.86 (d, J= 7.1 Hz, 3H). 25 "Example 5: (S)-4-Z,5-8-Hl-4-4 HE-9- A E-3,14-— AR-3,4,12,14- PO SL-1 A-AEB F# [3',4" 6,7] 5] BE Synthesis of F+[1,2-b] EMK-11-FA BB CSTI-F-01) cl OH OOO = N 1) TsOAg,DMSO NN On Hom: F How 0 STI-A-4 STI-F-01 The compound STLA-4 (250 mg, 0.58 mmol) AMT FF EMER AR (200 mg, 0.70 mmol) ¥% from Example 1 was stirred overnight at room temperature under nitrogen protection in 20 mL DMSO, and then heated and stirred at 90*C for 3 hours. After LCMS monitoring showed the formation of aldehyde 30", a portion of the reaction solution was taken for low-pressure reverse preparation (alkali mobile phase), oxidized in air, and lyophilized to obtain 50 mg of the title compound (STI-F-01), AA.LCMS, Rt = 1.25 min; MS calcd. value: 424.1; MS found value: 425.2 [M+H]^{+}. 53 WO 2024 / 255740 PCT / CN2024 / 098491 1H NMR (400 MHz, DMSO-d6) δ 8.85 (d, J = 8.5 Hz, 1H), 7.84 (d, J= 11.0 Hz, 1H), 7.29 (s, 1H), 6.50 (s, 1H), 5.42 (s, 2H), 5.27 (s, 2H), 2.47 (s, 3H), 1.92 — 1.80 (m, 2H), 0.88 (t, J=7.4 Hz, 3H). Example 6: Synthesis of (S)-7-Z.F&-7-FEFE-10,13-— Al-1 1H-[1,3] = AAR CFF [4,5-g]HS FF [3',4' 26,7] 5 1S 5 3£]1,2-b]E0K-8,11(7H)-— BA CSTI-EOS) and Example 7: (S)-7- ZFe-7-FEEE-1 4-(4-F4E TJ H8)-10,13-= A - 11 -[1,3] = BAR CFF [4,5-g] Ot F£[3',4':6,7] SERIF [1 2-b] -AMK-8,11(7H)-— Ad CSTI-E) Br OH 、 J o 、 Nitric acid O ~ FeSO4·7H2O O ss 6a 6b 6c 6d 6 OH N fap TORS ho 一 -一 ey te Ho = ° NY 《 LN ne CC OH ° VN N\4 CH3COOH, toluene, 80°C Hore LA -Ruttpych HEIR, cream A soon’ STI-E05 Bet sve 玉 ° Step 1: Under nitrogen atmosphere, add compound 6a (1 g, 7.24 mmol) into a reaction flask, then add 15 mL DMF 10 to dissolve it. Add potassium carbonate (3.84 g, 10.86 mmol) and dibromomethane (1.89 g, 10.86 mmol). Stir at FMB 110 for 2.5 H. TLC (PE:DCM = 1:1, product Rf= 0.7) monitors the completion of the reaction, and stop the reaction. Prep-TLCPurification yielded 972 mg of compound 6b, with a yield of 89.4%. Step 2: Compound 6b (972 mg, 6.48 mmol D) was added to a reaction flask, followed by 20 mL of nitric acid. The reaction was carried out at room temperature for 0.5 hours under fluorine protection. TLC (PE:EA = 5:1, product Rf = 0.5) detected the reaction results. After the reaction was complete, ice water was added to the reaction system. 15 HAAN, SAREE, BELAY 6c, a yellow solid of 1.053 g, yielded 83.3%. 1H NMR (400 MHz, CDCHD) 8 10.22 (s, 1H), 7.45 (s, 1H), 7.26 (s, 1H), 6.14 (s, 2H). Step 3: Compound 6c (7.0 g, 35.87 mmol D) was added to 350 mL of 5% HAAN, SAREE, BELAY 6c. In an ethanol-water solution, heat at 100°C; add FeSO4·7H2O (70 g, 251.09 mmol) to 350 mL of water, heat at 100°C for 2 minutes, and add to the reaction system. Slowly add 91.0 mL of concentrated nitrogen water. Heat at 100°C for 15 minutes. After the reaction is complete, return to room temperature, filter, cool in an ice bath, and precipitate 4.5 g (OA 6d), PK 76.0%. LCMS (254 nm) purity 96.61%, Rt = 1.40 min; MS calculated value: 165.0; MS measured value: 166.1 [M+H]'. 'H NMR (400 MHz, CDCH) 8 9.61 (d J= 0.6 Hz, 1H), 6.82 (s, 1H), 6.27 (s, 2H), 6.14 (s, 1H), 5.93 (s, 2H). Step 4: Compound 6d (300 mg, 1.82 mmol / D) and compound lc (360 mg, 1.37 mmol / D) were suspended in 4 mL of carboxylic acid, and 4 mL of ZR. AAP was added and heated overnight at 80°C. After the reaction was completed by LCMS monitoring, the mixture was cooled to room temperature, filtered, and washed with acetone to obtain 45 mg of compound STIL-E05.Pale yellow solid, yield 45.9%. LCOMS (254 nm) pure FE 99.62%, Rt = 1.49 min; MS calculated value: 392.1; MS measured value: 393.2 [M+H]. !IH NMR (400 MHz, DMSO-ds) 8 8.46 (s, 1H), 7.51 (s, 2H), 7.25 (s, 1H), 6.50 (s, 1H), 6.28 (dd, J = 1.4, 0.7 Hz, 2H), 5.41 (s, 2H), 5.21 (s, 2H), 1.92 — 1.80 (m, 2H), 0.87 (t, J=7.3 Hz, 3H). 54 WO 2024 / 255740 PCT / CN2024 / 098491 Procedure $: Compound STLE05 (100 mg, 0.26 mmol), IE JM (58 mg, 0.78 mmol), =e nea (WAT ANIK) GYA.6 mg, 0.0026 mmon D and 4,5,6,7-tetrafluoro-1-hydroxy-1)3-benzo[d]

[12] iodooxyl-3(1H)-one (175 mg, 0.52 mmol JIA RGR, AL 5S mL hexafluoroisobutyl alcohol (HFIP) mixed, bubbled under nitrogen for 1 hour, heated to 70"C under nitrogen protection, irradiated with a 23W energy-saving lamp, stirred for 65 hours. LCMS monitoring showed approximately 20% conversion, at which point the reaction was stopped. The product was then prepared by low-pressure reverse reaction (alkali mobile phase), lyophilized to obtain 20 mg of compound STI-E as a brownish-yellow solid. LCMS (254 nm) showed purity 94.36%, Rt = 1.75 min; MS calculated value: 464.2; MS measured value: 465.2. [M+H]-. ]!IH NMR (400 MHz, DMSO-de) 8 7.55 (s, 1H), 7.50 (s, 1H), 7.30 (s, 1H), 6.51 (s, 1H), 6.36 (t, J= 6.2 Hz, 1H), 6.29 (s, 2H), 5.42 (s, 2H), 5.26 (s, 2H), 3.92 — 3.82 (m, 2H), 3.79 (d, J= 6.7 Hz, 2H), 1.92 — 1.81(m, 4H), 1.38 — 1.26 (m, 2H), 0.84 (t, J= 6.8 Hz, 3H). 10 SCH 8: (S)-4-Z,F-4-FAIL-8-Sl-9- A E-1,12-— A -1 4-H SF [3',4': 6,7] 5] SSE [1,2-b] EM -3,14(4HD- Synthesis of dione (STI-F-8) NH2OH,HCl CClzCH(OH), fe) H2SO4 ? H2O> KOH COOH Th, HGcvNasouho Domo 8 see. ho 2oe la 8a Oo 8b 8c N LiAIH, OH MnO, So i 20 -AN f THF TA. DCM AL. TsOH, toluene,130°C eeate. 8d 8e F HO 20 STI-F-8 Step 1: i NaoSOu (22.7 g, 159.81 mmol) RK AR (3.0 g, 17.58 mmol) JA=Fie +H, Ho 15 A67mL water, then add compound 1a (2.0 g, 15.98 mmol D, carboxyamine hydrochloride (4.4 g, 63.92 mmol) All 25 mL 1M hydrochloric acid, and heat and stir overnight at 80*C under nitrogen protection. After the reaction is complete, filter, dry the filter cake at 50*C to obtain 2.9 g of compound 8a, as a yellow solid, yield 92.5%. LCMS (254 nm, purity 97.76%, Rt = 1.522 min; MS calculated value: 196.1; MS measured value: 197.1 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 58 12.21 (s, 1H), 10.28 (s, 1H), 7.64 (s, 1H), 7.61 (dd, J= 12.3, 2.0 Hz, 1H), 7.36 (dd, J= 8.3, 2.0 Hz, 1H), 7.22 (t, J=8.6 Hz, 1H), 2.19 (d, 20 J=1.9Hz, 3H). Step 2: Compound 8a (2.9 g, 14.79 mmol WF) was heated and stirred in 10 mL concentrated sulfuric acid at 80°C for 3 hours. The reaction mixture was monitored by LCMS until the reactants were completely reacted.The system was cooled to room temperature, and 200 mL of KK, WE, and YEG were added to the reaction system under ice bath conditions. The mixture was washed with water until the filtrate was neutral. The filter cake was dried at 0°C to obtain 2.07 g of compound 8b as an orange-yellow solid, with a yield of 78.2%. ¹H NMR (400 MHz, DMSO-ds): 11.06 (s 1H), 7.51 (d, J = 7.7 Hz, 1H), 6.69 (d, J = 9.8 Hz, 1H), 2.16 (d, J = 2.2 Hz, 3H). Step 3: Compound 8b (2.07 g, 11.6 mmol) was dissolved in 20 mL of 7K URIAF and 90 mg of 1 Hz O₂ was slowly added dropwise. The mixture was stirred at room temperature for 3 hours. After the reaction was complete, the pH was adjusted to 4-6 with hydrochloric acid in an ice bath, the mixture was washed with JE, and the filter cake was dried at 50°C to obtain 660 mg of compound 8c, which was a yellow solid. LCMS (254 nm, purity 94.31%, Rt = 1.065 min; MS calculated value: 169.1; MS measured value: 170.1 [M+H]'. 'H NMR (400 MHz, DMSO-de) 8 30 7.61 (d, J= 9.2 Hz, 1H), 6.49 (d, J= 12.3 Hz, 1H), 2.07 (s, 3H). Step 4: AR I PULA 10 mL PUA, nitrogen protection, ice bath cooling to 0"C, then add LiAlH4 (352 mg, 9.28 mmoloD), slowly add compound 8c (100 mg, 0.6 mmol) fH) PU APRIR (LO mD) deep solution, after addition, naturally 55 WO 2024 / 255740 PCT / CN2024 / 098491 KAP Bin, WARM. RMAKI, UIE RA 10°CLUR, RMIMAGERK, GHRZABAERL, saturated and washed with sodium oxide, dried with anhydrous sodium sulfate, filtered, vortexed and dried to give 510 mg KAW 8d as a brownish-yellow solid, yield 82.9%. LCMS (254nm) #4 FF 88.5%, Rt= 1.375min; MS calculated value: 15.1; MS measured value: 156.1 [M+H]. 1H NMR (400 MHz, DMSO-de) 8 6.90 (d, J= 9.0 Hz, 1H), 6.36 (d, J= 12.2 Hz, 1H), 4.94 5 (d, J= 5.5 Hz, 3H), 4.32 (d, J= 4.7 Hz, 2H), 2.05 (s, 3H). Step 5: Add MnO; (1.5 g, 17.11 mmol) to 10 mL of dioxane containing compound 8d (510 mg, 3.29 mmol) JAF and stir overnight at room temperature. After the reaction was completed, the solution was washed, vortexed, and purified by column chromatography to obtain 464 mg of compound 8e, AYR He [El AK, PERE 92.2%. LCMS (24nm) purity 84.82%, Rt = 1.787 min; MS calculated value: 153.1; MS measured value: 154.1 [M+H]*. 'H NMR (400 MHz, CDCls) 5 9.69 (s, 1H), 7.21 (d, J= 8.4 Hz, 1H), 6.23 10 (d, J= 11.6 Hz, 1H), 6.17 — 5.88 (m, 2H), 2.11 (d, J= 1.8 Hz, 3H). Step 6: Compound ge (466 mg, 3.04 mmol) ALLA Y 1c (600 mg, 2.28 mmol / L) was dissolved in 28 mL HK, heated to 130°C under nitrogen protection, and stirred under reflux for 30 minutes after water separation. 2) ela, DUA TsOH (131 mg, 0.76 mmol / L) was added and stirred continuously. The reaction was monitored by LCMS. After the reaction was completed, the system was cooled to room temperature, filtered, washed with acetone, and the filtrate was concentrated and prepared under medium and low pressure in reverse (alkali mobile phase). The target component was collected, lyophilized, and 350 mg of the title compound (STI-F-8) was obtained as a 15" yellow solid, with a yield of 40.4%. LCMS (254 nm) showed a purity of 94.07%, Rt = 1.589 min; MS calculated value: 380.1; MS measured value: 381.1 [M+H]*. 'HNMR (400 MHz, DMSO-d₆) δ 8.61 (s, 1H), 8.05 (d, J = 8.5 Hz, 1H), 7.88 (d, J= 10.9 Hz, 1H), 7.32 (s, 1H), 6.53 (s, 1H), 5.42 (s, 2H), 5.25 (s, 2H), 2.48 (s, 3H), 1.92 — 1.80 (m, 2H), 0.88 (t, J = 7.4 Hz, 3H). Example 9: Synthesis of (S)-N-(4-ethyl-4-hydroxy-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]naphtho[1,2-b]isoquinolin-11-yl)-2-carboxyacetamide (STI-F2), and Example 10: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-methyl-11-amino-1,12-dihydro-14H-pyrano[3',4':6,7]naphtho[1,2-b]isoquinoline-3,14(4H)-dione (STI-F-03c-1) fe) fe] Cl O — N Ac₂O, Py sx ON 1) H₂O₂, AcOH, 70°C = N Hac 入 人 Je 一 一 ~- hac a4 eo = > ite eI N 40°C N . 2) (COCl)₂, DMF, 0°C~rt. N F HOw: “O F AcO = 0 F AcO STI-F-8 一 9a 9b , ⁿBuOK, Pd(OAc)₂, Xantphos me N 7 人 AgNO₃ ye 6 NH₂ aeete, - O 、 一 , N 2) HCl / THF F AcO*: 0 2) MeOH, LiOH III 一 F HO STI-F2 H₂N fe) = N LiOH et F HO STI-F-03c-1 Step 1: At room temperature, mix 3.0 mL of acetic anhydride and 2.4 mL of pyridine uniformly, add compound STI-F-8 (300 mg, 0.79 mmol), heat and stir at 40°C, monitor by LCMS. After completion of the reaction, add saturated NaCl, extract with dichloromethane,PCT / CN2024 / 098491 Phase, medium-low pressure reverse preparation (alkali mobile phase), collect target component, freeze dry, to obtain 117 mg of compound 9a as a yellow solid, yield 3.1%. LCMS (24 nm, purity 97.16%, Rt = 1.698 min; MS calculated value: 422.1; MS measured value (A: 423.2) [M+H]*. 'H NMR (400 MHz, CDCl) 8 8.23 ​​(s, 1H), 7.77 — 7.64 (m, 2H), 7.12 (s, 1H), 5.60 (d, J = 17.2 Hz, 1H), 5.33 (d, J = 17.2 Hz, 1H), 5.19 (t, J = 1.5 Hz, 2H), 2.46 (t, J = 1.4 Hz, 3H), 2.21 (dd, J = 13.9, 7.5 Hz, 1H), 2.15 (s, 3H), 2.08 (dd, J = 13.9, 7.5 Hz, 1H), 0.91 (t, J = 7.5 Hz, 3H). WWE 2: Compound 9a (480 mg, 1.14 mmol) was added to 5 mL of acetic acid, followed by 6 mL of 30% ammonia peroxide aqueous solution. The mixture was heated and stirred at 70°C for 3.5 hours, collected by DCM, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. 24 mL of DMF TARE, URS ENE 0°C was added, followed by slow dropwise addition of 1.5 mL of oxaloyl chloride. After the reaction was complete, the mixture was brought to room temperature and stirred. The reaction was monitored by LCMS. After 10 °C, the mixture was sterilized by adding water, collected by DCM, evaporated to dryness, and prepared by reverse mixing under medium and low pressure (alkali mobile phase). The target fraction was collected, lyophilized, and 420 mg of compound 9b was obtained as a pink solid, with a yield of 81.0%. LCMS (254 nm) purity 97.37%, Rt = 2.014 min; MS calculated value: 456.1; MS measured value: 457.0 [M+H]*. 4H NMR (400 MHz, DMSO-d6) 8.24 (d, J = 8.0 Hz, 1H), 7.98 (d, J = 10.6 Hz, 1H), 7.05 (s, 1H), 5.50 (d, J = 4.2Hz, 2H), 5.28 (d, J = 4.7 Hz, 2H), 2.54 (s, 3H), 2.22 (s, 3H), 2.18 — 2.12 (m, 2H), 0.91 (t, J = 7.4 Hz, 3H). 15 Step 3: ELE 9b (240 mg, 0.52 mmol). t-BuOK (128 mg, 1.04 mmol). Pd(OAc)2 (16 mg, 0.07 mmol), Xantphos (30 mg, 0.05 mmol)#l — 7 FA Ae WZ He (104 mg, 0.58 mmol) ANAK IY, after purging with nitrogen 3 times, add 16 mL REF ASR, 100°CHIFAEEE 6 UM. HEP RDO, DOZAA Solution: Prepare by reverse process under medium and low pressure (alkali mobile phase), collect the target component, freeze dry, and obtain the coupling product. 90 mg of the coupling product was added to 15 mL of THF and 2 mL of 6M hydrochloric acid. The mixture was stirred overnight at room temperature. The reaction solution was concentrated, and the product was prepared by low-to-medium pressure reverse radiation (alkali flow 20" dynamic phase). The target component was collected, lyophilized, and 50 mg of LAY 9c was obtained as a pale yellow solid. The overall yield of the two steps was 21.7%. LCMS (254 nm) purity 90.36%, Rt = 1.730 min; MS calculated value: 437.1; MS measured value: 438.2. [M+H]'. 'H NMR (400 MHz, DMSO-de) 8.25 (d, J = 8.3 Hz, 1H), 7.58 (d, J = 11.4 Hz, 1H), 7.38 (s, 2H), 6.87 (s, 1H), 5.46 (s, 2H), 5.08 — 4.97 (m, 2H), 2.43 (s, 3H), 2.20 (s, 3H), 2.15 — 2.07 (m, 2H), 0.90 (t, J = 7.4 Hz, 3H). Step 4: Add 9gc (50mg, 0.01 mmol) of compound and DMAP (1mg, cab) to the reaction flask under nitrogen protection. Then add supernatant DMF (1.5ml), followed by DIPEA.(29 mg, 0.023 mmol D, 31 mg, 0.023 mmol, reacted at room temperature for 2 hours, then heated to SS$"C and reacted for 3 hours, then stirred at room temperature overnight. LCMS detected that the reaction was about 0%, the reaction was stopped, the system was concentrated to dryness under reduced pressure, methanol (4 mL) and LiOH (50 mg) were added, stirred at room temperature for about 30 min, the pH was adjusted to 2-3 with acetic acid, the system was concentrated to dryness, prepared by medium-low pressure alkaline reverse phase high pressure, lyophilized, to obtain 30 " to compound STILF2, a pale yellow solid of about 7.5 mg; the remaining compound gc in the reaction system was hydrolyzed with LiO to obtain compound STI-F-03c-1, a pale yellow solid of about 8.3 mg. Compound STI-F2: LCMS (254 nm) purity 97.89%, Rt = 1.51 min; MS Calculated value: 453.1; MS measured value: 454.2 [M+H]*. 1H NMR (400 MHz, DMSO-ds) 8 10.65 (s, 1H), 8.13 (d, J = 8.0 Hz, 1H), 7.90 (d, J = 10.8 Hz, 1H), 7.32 (s, 1H), 5.81 (br, 1H), 5.43 (s, 2H), 5.12 (s, 2H), 4.30 (s, 2H), 1.92-1.81 35 (m, 2H), 0.88 (t, J = 7.6 Hz, 3H). Compound STI-F-03c-1: LCMS (254nm)#E FF 97.67%, Rt = 1.60 min; MS calculated value: 395.1; MS measured value: 396.2 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 5 8.76-8.58 (m, 2H), 8.38 (d, J = 8.0 Hz, 1H), 7.60 (d, J = 10.84 Hz, 1H), 7.46-7.42 (m, 1H), 6.61 (br, 1H), 5.44 (s, 2H), 5.05 (s, 2H), 2.44 (s, 3H),1.93 — 1.78 (m, 2H), 0.87 (t, J = 7.2 Hz, 3H). Example 11: Synthesis of (S)-4-ethyl-4-hydroxy-8-amino-1L-pentyl-112-dioxo-14H-pyteno[3.4':6,7]oxazino[1,2-b]SUK-3,14(4H)-—A (STLG1) fe) ol — a "TL o AlCls ON i F OH EtsN, DCM F oR PhNOg, 150°C [oeee dia 11b 11c 1) NH2Boc, Pd2(dba), O Xantphos,CszCOs% O Tf,O, Pyridine oN we Dioxane. 80°C ON wee Fe, NH,Cl DCM, 0°C Fc ott 2) 50% TFA / DCM NH, EtOH / H2O, 80°C 11d tle oO “coo ° HG 1c O H2N 1 person—_ F NH2 TsOH, toluene, reflux F 、 How 11f STI-G1 Step 1: Under nitrogen and ice bath conditions, compound 11a (500 mg, 3.18 mmol / D) was dissolved in 30 mL DCM. Hexanoyl chloride (0.67 mL, 4.77 mmol / D) and triethylamine (0.88 mL, 6.36 mmol / D) were added to the reaction flask with stirring at 0°C. The reaction was continued at this temperature for 30 minutes. The reaction was confirmed to be complete by TLC (PE:EA=30:1, product R0.5). 50 mL of Sa 10" alkyl and 50 mL of water were added, and the reaction was repeated twice. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA = 10:1) to give 650 mg of compound 11b as a colorless liquid, yielding 80.9%. ¹H NMR (400 MHz, DMSO-ds) ¹⁵ 8.25 (t, J = 8.9 Hz, 1H), 7.54 (dd, J = 12.1, 2.4 Hz, 1H), 7.27 (dt, J = 9.1, 1.8 Hz, 1H), 2.62 (t, J = 7.4 Hz, 2H).1.64 (dd, J = 10.0, 4.8 Hz, 2H), 1.33 (q, J = 3.7 Hz, 4H), 0.89 (t, J = 6.9 Hz, 3H). 15 ER 2: BALA 11b (500 mg, 1.96 mmol) ¥4-F 4 mL TASER, aluminum oxide (290 mg, 2.16 mmol) was added under nitrogen protection and the reaction was carried out at 150°C. BSD 5 MY. TLC (PE:EA = 30:1, product Rf = 0.5) was used to detect the reaction. After the reaction was completed, the reaction system was cooled to room temperature and added to 0 mL of IN HCl in ice water. The mixture was stirred at room temperature for 30 minutes. Add 50 mL of ZAR ZARA, extract the aqueous phase twice with ethyl acetate, combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (PE:EA=200:1 to remove the nitro cosolvent, PE:EA=20:1 to pass the product). Compound 11c is a yellow solid, yield 60%. ¹H NMR (400 MHz, DMSO-d6) ¹²⁸ 12.66 (s, 1H), 8.54 (d, J = 9.0 Hz, 1H), 7.04 (d, J = 13.0 Hz, 1H), 3.06 (t, J = 7.2 Hz, 2H), 1.60 (dd, J = 8.5, 5.9 Hz, 2H), 1.33 — 1.28 (m, 4H), 0.89 — 0.85 (m, 3H). Step 3: Add pyridine (0.63 mL, 7.84 mmol AF) and THO (0.99 mL, 5.88 mmol) to the reaction system of compound 11c (500 mg, 1.96 mmol AF) in 20 mL SAP, UKIB RATER PA, and FRAY for 30 minutes. After the reaction is complete, add 20 mL of dioxane to the reaction system, extract with water, and extract the aqueous phase twice with 10 mL of dioxane. Combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and 58 WOThe brown crude product obtained from 2024 / 255740 PCT / CN2024 / 098491 was purified by silica gel column chromatography (PE:EA=20:D, after bipolar chromatography) to give 650 mg of compound 11d as a yellow solid, with a yield of 85.7%. ¹H NMR (400 MHz, CDCH) 5 8.55 (d, J= 7.9 Hz, 1H), 7.36 (d 7= 9.9 Hz, 1H), 2.96 (t, J= 7.3 Hz, 2H), 1.75 (dd, J= 9.1, 5.6 Hz, 2H), 1.36 (q, J= 3.9 Hz, 4H), 0.93 (d, J= 6.8 Hz, 3H). Step 4: Compound 11d (1.12 g, In a nitrogen-protected system, 28 mL of 1,4-dioxane was prepared by adding butyl carbamate (680 mg, 5.80 mmol), Xantphos (336 mg, 0.58 mmol), Pd2(dba)s (266 mg, 0.29 mmol), and FUE HA (1.89 g, 5.80 mmol). The reaction was monitored by TLC (PE:EA = 10:1, product Rf = 0.6). After the reaction was complete, the system was cooled to room temperature, diluted with 20 mL of ethyl acetate, and water was added. The aqueous phase was then diluted twice with 10 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting brown crude product was purified by silica gel column chromatography (PE:EA = 50:1). After rotary evaporation, 440 mg of a yellow liquid was obtained. The yellow liquid was dissolved in 12 mL of 50% TFA / DCM JAK, FIL DY for 30 minutes. The reaction was detected by TLC (PE:EA = 5:1, product Rf = 0.3). After the reaction was complete, 20 mL of dioxane was added to the reaction system for dilution, followed by water collection. The aqueous phase was collected twice more with 10 mL of dioxane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting brown crude product was subjected to silica gel column chromatography.Purified by precipitate (PE:EA=5:UD), and after rotation and drying, 260 mg of compound 1 eI was given as a pale yellow solid, yield 35.3%. 15 LCMS (254 nm) Rt = 2.108 min; MS calculated value: 254.1; MS measured value: 253.1 [MH] .'H NMR (400 MHz, DMSO-ds) 8.64 (d, J = 8.7 Hz, 1H), 8.53 — 7.43 (m, 2H), 6.67 (d, J = 14.4 Hz, 1H), 3.01 (t, J = 7.3 Hz, 2H), 1.59 (t, J = 7.2 Hz, 2H), 1.31 (dg, J = 7.6, 3.4 Hz, 4H), 0.92 — 0.83 (m, 3H). WHE 5S: Compound 11le (200 mg, 0.79 mmol) was added to a 12 mL mixture of ethanol:water = 3:1 under nitrogen protection. Iron powder (176 mg, 3.16 mmol) and FE (44 mg, 0.79 mmol) were added to the reaction system. BHAA 3 times, 20 $80 © ERIN. TLC (PE:EA = 5:1, product Rf = 0.2) to detect the reaction. After the reaction was completed, the reaction was cooled to room temperature and soluble in the reaction solvent. The crude product was purified by silica gel column chromatography (PE:EA = 2:1). After evaporation to dryness, 160 mg of compound 11f was obtained as a brown solid, with a yield of 90%. LCMS (254 nm) Rt = 2.189 min; MS calculated value: 224.1; MS measured value: 225.2 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 5 7.25 (d, J = 10.1 Hz, 1H), 6.70 (s, 2H), 6.45 (d, J = 13.5 Hz, 1H), 4.39 (s, 2H), 2.78 (t, J = 7.4 Hz, 2H), 1.57 (t, J = 7.3 Hz, 2H), 1.30 (tt, J = 5.9, 3.2 25 Hz, 4H), 0.87 (t, J = 6.8 Hz, 3H). Step 6: Compound 11f (20 mg, 0.09 mmol) FILA Ic (24mg, 0.09 mmol of FF were dissolved in 1 mL of anhydrous methanol. Under nitrogen protection, the mixture was refluxed with a water separator for 4 hours. DABCO (4 mg, 0.023 mmol) was added, and the mixture was continuously stirred and refluxed for 3.5 hours. After the reaction was detected to be completed by TLC (DCM:MeOH=30:1, product Rf = 0.4) and LCMS, the reaction system was cooled to room temperature, filtered, washed with acetone, and the crude product was separated and purified by preparative plate to obtain the title compound (STI-G1H) as a yellow solid, 5 mg. LCMS (254nm) Rt = 1.839 min; Calcd mass: 451.2; Found mass: 452.3 [M+H]+. 1H NMR (400 MHz, DMSO-d6) δ 7.74 (d, J= 12.3 Hz, 1H), 7.30 (d, J=9.5 Hz, 1H), 7.19 (s, 1H), 6.46 (s, 1H), 6.06 (s, 2H), 5.40 (s, 2H), 5.21 (s, 2H), 3.00 (t, J = 8.1 Hz, 2H), 2.05 — 1.94 (m, 2H), 1.91 — 1.79 (m, 2H), 1.73 — 1.62 (m, 2H), 1.48 — 1.40 (m, 2H), 0.90 (t, J= 6.0 Hz, 3H), 0.87 (t, J = 5.9 Hz, 3H). Example 12: Synthesis of (S)-4-ethyl-4-hydroxy-8-fluoro-9-nitro-11-pentyl-1,12-dihydro-14H-pyrano[3'',4':6,7]isoquinolino[1,2-b]quinazoline-3,14(4H)-dione (STI-G03) WO 2024 / 255740 PCT / CN2024 / 098491 Compound 11e (20 mg, 0.078 mmol) and 1c (20 mg, 0.078 mmol) were added to 1 mL of toluene. Under nitrogen protection, the mixture was refluxed with a water separator for 30 minutesbell. Add p-toluenesulfonic acid (3.5 mg, 0.02 mmol / D), and continue reflux with stirring for 3.5 hours. After the reaction was complete, the reaction system was cooled to room temperature (5 °C), filtered, washed with acetone, and the crude product was purified by preparative chromatography to obtain the title compound as a yellow solid (8 mg, yield 21%). LCMS (254 nm) purity 93.78%, Rt = 2.01 min; MS calculated value: 481.2; MS measured value: 482.2 [MH+HHT]. 1H NMR (400 MHz, DMSO-ds) 9.05 (d, J = 7.9 Hz, 1H), 8.26 (d, J = 12.2 Hz, 1H), 7.38 (s, 1H). 6.57 (s, 1H), 5.45 (s, 2H), 5.35 (s, 2H), 3.30 — 3.25 (m, 2H), 1.92 — 1.82 (m, 2H), 1.72 (t, J= 7.7 Hz, 2H), 1.49 — 1.44 (m, 2H), 1.38 (t, J= 7.1 Hz, 2H), 0.88 (d, J= 8.1 Hz, 6H). 10 Example 13: (S)-9-Ye-4- 2, 3E-8-F-4-PEHE- 1 1- A te FA K-11 2-2-1 4-H FF [3,4]: 6,7] 51 WE F2[1,2-b] Synthesis of EWK-3,14(4H)-— BW (STI-G2-06c) 0 er ee es Or AP 00, aaa Cte” SES en eg Ry Step 1: TERA AUGER AEE FS ALA 70 mL DCE and 1M =a) DCM solution 15 (12.5 mL, 12.5 mmoloD, then add compound 13a (3.0 g, 15.6 mmol), O°CHEFE 10 44. ARAEFEVIOA BIAS ZSRU416 mg, 18.7 mmoloD and aluminum trichloride (2706 mg, 20.3 mmoloD, then slowly raise the reaction system to room temperature. Stir at room temperature for 10 minutes, then reflux and stir for 39 hours.After the reaction was complete, the system was added to 100 mL of ice water, followed by 40 mL of 2 M HCl, and stirred at room temperature for 1 hour. Add 100 mL of dioxane to extract the organic phase, dry it with anhydrous sodium sulfate, concentrate under reduced pressure, and prepare the residue by reverse high-pressure neutralization (alkali mobile phase). After lyophilization, 20-11 g of compound 13b was obtained as a yellow solid powder, with a yield of 26.2%. LCMS (254 nm, purity 100%, Rt = 1.93 min; MS calculated value: 264.9; MS measured value: 263.9 [MH]). ¹H NMR (400 MHz, DMSO-d6O) 5 8.03 (d J = 7.9 Hz, 1H), 7.56 (s, 2H), 6.74 (d, J = 11.6 Hz, 1H), 5.05 (s, 2H). Step 2: Add 250 mg of compound 13b (0.94 mmol AF) to 5 mL of 0.94 mmol AF. OAR. AER, IA (94 uL, 1.03 mmoD and triethylamine (261 wL, 1.88 mmol), stirred overnight at room temperature, TLC (PE:EA = 5:1, product Rf = 25 0.3) was used to detect the reaction. After the reaction was complete, the mixture was washed with water, dried, and purified by pre-TLC to give 218 mg of compound 13c as a pale yellow solid, with a yield of 75.7%. 1H NMR (400 MHz, DMSO-ds) 5 8.08 (d, J = 8.0 Hz, 1H), 7.53 (s, 2H), 6.71 (d, J = 11.5 Hz, 1H), 3.90 (s, 2H), 2.47 (d, J = 7.3 Hz, 2H), 1.54 (h, J = 7.3 Hz, 2H), 0.91 (t, J=7.3 Hz, 3H). Step 3: Add 10 mL of compound 13c (187 mg, 0.61 mmol AMLG 1c (121 mg, 0.46 mmol) J8-F ACK AE and nitrogen protection, and reflux for 30 eH using a water separator. Add DAT HAR TRFR (11 mg, 0.06 mmol D, and continue reflux with stirring for 60 rpm). WO 2024 / 255740PCT / CN2024 / 098491, 40° F for 4 hours. After LCMS detection showed the reaction was completed, the reaction system was cooled to room temperature, filtered, washed with acetone, and purified by Pre-TLC (DCM:MeOH = 15:1) to obtain 230 mg of compound STI-G2-06c as a yellow solid, with a yield of 94.0%. Rt = 2.09 min; MS calculated value: 532.1; MS observed value: 533.1 [M+H]. 1H NMR (400 MHz, DMSO-d6) δ 8.79 (d, J = 7.5 Hz, 1H), 8.13 (d, J= 9.8 Hz, 1H), 7.33 (s, 1H), 6.54 (s, 1H), 5.44 (s, 2H), 5.33 (s, 2H), 4.45 (s, 2H), 2.05 — 1.95 (m, 2H), 1.92 — 1.81 (m, 2H), 1.63 — 1.52 (m, 2H), 0.93 — 0.88 (m, 3H), 0.87 — 0.82 (m, 3H). Example 14: Synthesis of (S)-9-Rk-4-Z,5k-8-F-4-FE-11-H FHE-1, 12-2 S-14-S(3,4: 6,7] 5] WEBS IE 1,2-b] EMK-3,14(4H)--Be (STI-G2) Boc-NHp, Pd(AcO)2, Cs2CO3, Xantphos S oN i TFA / DCM ry penta 人TO RD -~ HN AN i re) F " Ho": 0 F HoT: 0 F Ho" = © 10 STI-G2-06c 14a STI-G2 Step 1: Under nitrogen protection, the compound STI-G2-06c (53 mg, 0.10 mmol) from Example 13 and Rk AAT H (23 mg, 0.20 mmol) were added to 5 mL of anhydrous dioxane. Under nitrogen protection, cesium carbonate (65 mg, 0.20 mmol), Xantphos (5.8 mg, 0.01 mmol) and palladium acetate (2.2 mg, 0.01 mmol) were added, followed by fHPEIIS], the mixture was heated to 100°C and reacted for 2.0 hours. TLC (DCM:MeOH = 15:1, 77-49After the reaction was completed (Rf = 0.5), the system was concentrated to dryness under reduced pressure. The organic phase was concentrated using a 15 GBB WB AUZKARER filter to obtain 40 mg of compound 14a as a yellow solid, yield 70.5%. LCMS (254 nm) purity 98.42%, Rt = 2.07 min; calculated value: 369.2; measured value: 570.3 [M+H]". Step 2: Compound 14a (65 mg, 0.11 mmol) was added to 2.0 mL DCM, along with 0.6 mL TFA, and stirred at room temperature for 0.5 hours under nitrogen protection. The reaction was stopped, and the mixture was concentrated to dryness under reduced pressure. The mixture was then prepared by reverse mixing under medium and high pressure (acidic mobile phase), and dried to obtain 10 mg of compound STI-G2 as a reddish solid, yield 18.7%. LCMS (254 nm) purity 98.48%, Rt = 2.07 min; calculated value: 369.2; measured value: 570.3 [M+H]". 1.75 min; 20 MS Calculated value: 469.2; MS measured value: 470.2 [M+HHT 。 IH NMR (400 MHz, DMSO-ds) 8 7.76 (d, J = 12.3 Hz, 1H), 7.35 (d, J = 9.6 Hz, 1H), 7.20 (s, 1H), 6.48 (s, 1H), 6.16 (s, 2H), 5.41 (s, 2H), 5.24 (s, 2H), 4.19 (s, 2H), 2.58 (t, J = 7.2 Hz, 2H), 1.91 — 1.80 (m, 2H), 1.65 — 1.56 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H), 0.87 (t, J = 7.3 Hz, 3H). 25” Example 1 S:, (S)-10-cyclopropyl-4-ethyl-8-amino-4-hydroxy-11-(4-hydroxybenzyl)-9-methyl-1,12-dioxo-14H-pyridineFF [3',4':6,7] 95] BRRSIF [1,2-b] ZEMK-3,14(4H)-—Ad (CSTI-D) Synthesis 61 WO 2024 / 255740 PCT / CN2024 / 098491 ioe . arkig~ OP opus 80 oe ke Sd x ead _ TFA eed Tk Ki LEBER,KPO, bed to ONO set FA o poM O Pd(dpphCl, dioxane 0 Ts0H.H,0, toulene aw h Step 1: At room temperature, compound 1S3a (10.0 g, 80.0 mmol JAF) was added to 100 mL DCM, 100 mL methanol was added, and then Br (32.0 g, 200.0 mmol D) was added dropwise to the system. After the addition was complete, the reaction was carried out at room temperature for two hours. After the reaction was completed, the system was concentrated to dryness under reduced pressure, and then 1M sodium thiosulfate aqueous solution (100 mL) and 100 mL ethyl acetate were added. The pH was adjusted to 7-8 with saturated sodium thiosulfate and NaHCO3. The aqueous phase was washed with EA (100 mL x 2). The organic phases were combined and washed successively with 100 mL of 1M sodium thiosulfate aqueous solution and saturated brine. Then the mixture was dried with anhydrous Na2SO4 and concentrated to dryness under reduced pressure. Purification by column chromatography (PE) yielded 20.0 g of compound 1Sb as a pale purple solid, yield 89.3%. ¹H NMR (400 MHz, DMSO-ds) ¹⁵ 7.44 (d, J = 8.8 Hz, 1H), ¹⁵ 5.16 (s, 2H), ¹⁵ 2.21 (d 7 = 2.4 Hz, 3H). Step 2: TsOH (40.0 g, 232.6 mmol), JA MeCN (240.0 mL), AIL 10°C, MALE 10 4 15b (20.0 g, 70.8 mmol); NaNO₃ (9.8 g, 141.6 mmol) & KI (29.4 g, 141.6 mmol) HEA F 7k (60.0 mL) were added dropwise to the reaction system. After the addition was complete, the system was allowed to return to room temperature naturally for reaction 1a, PIER response. The system was poured into 200 mL of water, and the pH was adjusted to approximately 9 with saturated NaHCO₃. Then, 1M sodium thiosulfate aqueous solution was added until the system no longer decolorized. The organic phases were combined with EA (200.0 mL x 3)2E HL, dried under reduced pressure, concentrated to dryness, and subjected to column chromatography.Purification (PE) yielded 18.6 g of compound 1$c as a white solid, yield 67.1%. ¹H NMR 15 (400 MHz, DMSO-ds) 7.79 (d, J = 9.2 Hz, 1H), 2.26 (d, J = 2.4 Hz, 3H). Step 3: Compound 1$c (5040 mg, 12.8 mmol) was reacted at -30°C for 1.5 h, then DMF (3083 mg, 42.2 mmol) was added dropwise to the system. After addition, the system was allowed to return to room temperature and reacted for 1.5 h. After the reaction was completed by TLC (PE:EA = 20:1, product Rf = 0.3), 50.0 mL of saturated 20" NH4Cl aqueous solution was added to the system to extinguish the reaction. The mixture was then eluented with EA (50.0 mL x 3), and the organic phases were combined, dried over anhydrous Na2SO4, concentrated to dryness under reduced pressure, and purified by column chromatography (PE-PE:EA = 100:1) to give 3.5 g of compound 15d as a pale yellow solid, with a yield of 79.8%. ¹H NMR (400 MHz, DMSO-ds) 10.14 (s, 1H), 7.32 (d, J = 12.6 Hz, 1H), 2.29 (d, J = 2.8 Hz, 3H). Step 4: Compound 15d (2959 mg, 10.0 mmol / L) was added to a three-necked flask. Under nitrogen protection, DCE (30.0 mL) was added to dissolve the precipitate, followed by the sequential addition of ethylene glycol (3724 mg, 60.0 mmol / D), triethyl orthoformate (1630 mg, 11.0 mmol / D), and TsOH 25 (86 mg, 0.5 mmol). The mixture was incubated at [ARDY] for 3 hours. The reaction was monitored by TLC (PE:EA = 20:1, product Rf = 0.29). After the reaction was complete, the reaction was stopped, and the system was allowed to cool naturally to room temperature. 50 mL of DCM was added to dilute the reaction solution, and the mixture was extracted with 80 mL of saturated Na2CO3 aqueous solution.The organic phase was washed with 80 mL of saturated brine, dried over anhydrous Na₂SO₄, and concentrated to dryness under reduced pressure to give 3.3 g of compound 1Se as a pale yellow solid, with a yield of 97.7%. ¹H NMR (400 MHz, DMSO-ds) ⁵ 7.33 (d, J = 62 Hz, 1H), ⁶ 6.42 (s, 1H), ⁴ 4.35–4.31 (m, 2H), ⁴ 4.09–4.06 (m, 2H), ⁴ 2.33 (d, J = 2.4 Hz, 3H). DER 5: 3.3 g of compound 1Se was added to a reaction flask at 10.0 mmol / L. Under atmospheric protection, methyl methacrylate (20.0 mL) was added. Next, dimethyl ketone imine (140 mg, 8.0 mmol), #BuOK (2356 mg, 21.0 mmol), Pd(OAc)2 (225 mg, 1.0 mmol), and Xantphos (579 mg, 1.0 mmol) were added sequentially to the reaction flask. The mixture was heated to 1005°C and reacted for 1 hour. After stopping the reaction, the system was filtered with diatomaceous earth. The filter cake was washed with EA, and the filtrate was concentrated to dryness under reduced pressure and purified by column chromatography (PE:EA = 400:1) to obtain 730 mg of compound 1Sf as a pale yellow solid, with a yield of 16.6%. Step 6: Compound 1Sf (730 mg, 1.67 mmol) was dissolved in 4.0 mL of THE, cooled to 0-3°C, and concentrated hydrochloric acid (11.0 mL, 116.8 mmol) was added dropwise to the system. After the addition was complete, the reaction was stopped after maintaining the temperature at 0-5°C for 10 minutes. The pH was adjusted to neutral with solid NaHCO3, and EA (20.0 mL) and 7K (20.0 mL) were added and collected. The aqueous phase was washed with EA (20.0 mL) and 7K (20.0 mL) for 10 minutes. The organic phases were combined, dried over anhydrous Na2SO4, concentrated to dryness under reduced pressure, and purified by Pre-TLC to give 130 mg of compound 15g as a yellow solid, with a yield of 33.7%. LCMS (254 nm) showed a purity of 92.14%, Rt= 2.024 min; MS calculated value: 231.0; MS measured value: 230.0 [MH]. 'H NMR (400 MHz, DMSO-da 8 10.39 (s, 1H), 6.51 (br, 2H) 6.31 (d, J= 11.2 Hz, 1H), 2.25 (d, J= 2.4 Hz, 3H). Step 7: 15 g of compound (130 mg, 0.56 mmol JIA RMIT, AAR, THE Aa BA 15 “0-$C, BF BUM 17D. FRIE BAY, [Al EA AEE, dry NazSO4, 10.39 (s, 1H), 6.51 (br, 2H) 6.31 (d, J= 11.2 Hz, 1H), 2.25 (d, J= 2.4 Hz, 3H). ... The solution was concentrated to dryness under reduced pressure and purified by Pre-TLC (PE:EA = 3:1) to give 130 mg of compound 151 as a brown oily liquid, with a yield of 54.6%. LCMS (254 nm, purity 98.60%, Rt = 2.174 min; MS calculated value: 425.1; measured value: 424.1 [MH]). ¹H NMR (400 MHz, DMSO-ds) ⁵ 7.26–7.17 (m, 2H), 6.93–6.84 (m, 2H), 6.44 (d, J = 12.0 Hz, 1H), 5.79 (d, J = 3.8 Hz, 20 Hz, 1H), 5.66 (s, 2H), 5.16 (dt, J = 8.7, 4.2 Hz, 1H). 4.34 (s, 2H), 3.74 (s, 3H), 3.36 (t, J = 6.2 Hz, 2H), 2.11 (d, J = 2.2 Hz, 3H), 1.87–1.74 (m, 1H), 1.55–1.46 (m, 5H). Step 8: Compound 1Si (130 mg, 0.31 mmol WF) was added to 4.0 mL of 2.7 ZEA, followed by 2-H EE FA (BX) (95 mg, 0.34 mmol), FRBNY. After 4 hours, the reaction was stopped, and the system was allowed to return to room temperature. The mixture was filtered through diatomaceous earth, the filter cake was washed with ethyl acetate, and the filtrate was reduced to a final volume under reduced pressure and then purified by Pre-TLC.(PE:EA=5:2), 130 25 mg of compound 15j was obtained as a brown oily liquid, yield 100.0%. LCMS (214 nm, purity 90.82%, Rt = 2.226 min; MS calculated value: 423.1; measured value: 422.1) [MH]-. !H NMR (400 MHz, DMSO-ds) 8 7.26-7.21 (m, 2H), 6.91-6.87 (m, 2H), 6.52 (d, J = 12.0 Hz, 1H), 5.35 (br, 2H), 4.36 (s, 2H), 3.74 (s, 3H), 3.39 (t, J = 6.4 Hz, 2H), 2.77 (t, J = 6.4 Hz, 2H). 7.2 Hz, 2H), 2.10 (d, J= 2.4 Hz, 3H), 1.68-1.63 (m, 2H), 1.61-1.55 (m, 2H). 30 Step 9: Dissolve compound 15j (130 mg, 0.31 mmol) in 1 mL DCM, add 3.0 mL of 50% trichloromethyl acetic acid solution to the IRA (IRA) dropwise, JHE, THI 0-5°C, stop the reaction after 20 minutes, redissolve in DCM, with saturated NaHCO3; adjust pH to 7, take, wash the aqueous phase with DCM (20.0 mD), combine the organic phases, dry with anhydrous Na2SO4, concentrate to dryness under reduced pressure, and purify by Pre-TLC (PE:EA=5:2). 80 mg of compound 15k was obtained, 772% (86.0%). LCMS (254 nm, purity 94.67%, Rt = 1.763 min; 35 MS, calculated value: 303.0; found value: 302.0 [MH]). ¹H NMR (400 MHz, DMSO-ds) ¹H NMR: 6.51 (d, J = 12.0 Hz, 1H), 5.35 (s, 2H), 4.34 (s, 1H), 3.40 (t, J = 6.4 Hz, 2H), 2.77 (t, J = 7.2 Hz, 2H), 2.11 (d, J = 2.0 Hz, 3H), 1.67–1.60 (m, 2H), 1.50–1.53 (m, 2H). Procedure10: Compound 15k (40 mg, 0.13 mmol) was added to AFF 1,4-dioxane (4.0 mD), followed by 63 WO 2024 / 255740 PCT / CN2024 / 098491 cyclopropylboronic acid (67 mg, 0.78 mmol), K3PO4 (165 mg, 0.78 mmol) & Pd(dppf)Cl2 (22 mg, 0.03 mmol). Under nitrogen protection, the mixture was heated to 90°C and reacted for 1.5 hours. The reaction was stopped by LCMS monitoring until the starting materials were completely consumed. The system was allowed to cool naturally to room temperature, filtered through silica gel, and the filter cake was washed with ethyl acetate. The filtrate was concentrated to dryness under reduced pressure and then purified by Pre-TLC (DCM:MeOH=20:1). #2 30 mg of compound 15k was a white solid. Yield 85.8%. Step 11: Compound 151 (21 mg, 0.08 mmol), compound 1c (40 mg, 0.13 mmol), and TsOH H2O (40 mg, 0.13 mmol) were added to a reaction flask. Methyl mercaptan was added, and the mixture was refluxed under choke protection for 1 hour. The reaction was then stopped, and the system was concentrated to dryness under reduced pressure. The mixture was prepared by reverse-phase reaction under medium-high pressure (acid mobile phase) and lyophilized to obtain 6.5 mg of compound STILD as a pale yellow solid. LC-MS (254 nm, purity 98.45%, Rt = 1.990 min; MS calculated value: 492.2; measured value: 493.1) [M+H]*. 'H NMR (400 MHz, DMSO-ds) 6.72 (s, 1H), 6.29 (s, 1H), 6.20 (d, J= 11.2 Hz, 1H), 10 6.13 (s, 1H), 4.56 (dd, J = 24.4, 16.0 Hz, 2H), 4.16-3.92 (m, 4H), 2.59-2.54 (m, 2H), 2.13 (d, J= 2.0 Hz, 3H), 2.12-2.08 (m, 1H) 1.95-1.85 (m, 2H), 1.84-1.61 (m, 4H), 0.99-0.88 (m, 2H), 0.75 (t,J = 7.2 Hz, 3H), 0.54-0.48 (m, 1H), 0.32-0.28 (m, 1H). The intermediate compound 15h was prepared as follows: 15h-1 15h-1 Magnesium granules (97 mg, 4.03 mmol) were added to a three-necked flask under nitrogen protection. 4.0 mL of THF and JA 1,2-diethane (2 drops, cat) were added, and the mixture was heated to initiate reaction. 15h-1 (914 mg, 3.36 mmol) was added to 2.0 mL of THF and the reaction mixture was reacted at 505°C for 30 minutes to obtain 15h HY THF YAR. Example 16: 2-Cyclodiyl-N-((S)-4-ethyl-8-fluoro-4- Synthesis of hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetraaza-1H-pyrano[L2-b]azolin-11-yl)-2-hydroxyacetamide (STILE) Nitrogen 25) URE A 3 mL anhydrous methyl methacrylate, stirred evenly, heated to 100°C, ABV 1.0 oz. TLC (DCM:MeOH=20:1, product R0.5) to detect the reaction after completion, the system was concentrated to dryness under reduced pressure, purified by column chromatography (DCM:MeOH=100:1), giving 17 mg of compound 16b as a yellow solid, yield 31.8%. LCMS (254 nm) purity 99.42%. Rt = 1.89 min; tt calculated: S33.2; measured: S34.2 [M+H]. Procedure2: Compound 16b (17 mg, 0.032 mmol) was added to DCM (1.0 mD) in four portions under atmospheric protection. 30 μL of STAB (28 mg, 0.140 mmolD) was added in each portion, and the reaction was carried out at room temperature for 1.5 hours. The reaction was monitored by TLC (DCM:MeOH = 20:1, product Rf = 0.3). After the reaction was complete, 10 mL of DCM and 10 mL of water were added to the system. The aqueous phase was collected and washed with DCM (10 mL x 2). The organic phases were combined and concentrated to dryness under reduced pressure. Methanol (3 mD) and lithium oxide (20 mg) were added, and the reaction was carried out at room temperature for 10 minutes. The pH was adjusted to 3-4 with acetic acid. Add a mixed solvent of dimethyl ether and methanol (DCM:MeOH = 10:1, 10 mL) and water (10 mL), wash with AER, KAA Sat cS AEE AYA (DCM: eOH = 10:1, 10 mL x 2), combine the organic phases, concentrate to dryness under reduced pressure, prepare by reverse mixing under medium and low pressure (acidic mobile phase), lyophilize to give 6 mg of compound STI-F, a pale yellow solid, yield 37.9%. LCMS (254 nm, purity 98.86%, Rt = 1.60 min; MS calculated value: 493.2; MS measured value: 494.2 [M+H]. 1H NMR (400 MHz, DMSO-d) 5 10. 60 (s, 1H), 8.07 (d J = 8.0 Hz, 1H), 7.92 (d, J = 10. ...8.07 (d J = 8.0 Hz 10.8 Hz, 1H), 7.33 (s, 1H), 6.52 (s, 1H), 5.91 (br, 1H), 5.42 (s, 2H), 5.11 (s, 2H), 3.95 (d, J = 6.8 Hz, 1H), 2.50 (s, 3H), 1.92-1.79 (m, 2H), 1.34-1.27 (m, 1H), 0.88 (t, J = 7.2 Hz, 3H), 0.62-0.49 (m, 4H). Intermediate compound 16a was prepared as follows: See HO OO H2N 16a-1 16aCompound 16a-1 (250 mg, 2.19 mmol THF (5 mL)) was mixed with triethylamine (0.61 mL, 4.38 mmol), and the mixture was protected with nitrogen and cooled to -20°C. A tetrahydroquinone solution of isopropyl oxyacetate (299 mg, 2.19 mmol) was added dropwise. After stirring at a constant temperature for 30 minutes, concentrated ammonia (1279 mg, 10.95 mmol) was added to the system. The reaction was stopped after 2 hours at room temperature. The system was concentrated to dryness under reduced pressure, and the organic phase was washed with saturated sodium carbonate aqueous solution and EA. The mixture was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. Pre-TLC (PE:EA = 2:1) showed that 92 mg of compound 16a was a white solid with a yield of 37.2%. ¹H NMR (400 MHz, DMSO-ds) 5 7.91 (s, 1H), 7.69 (s, 1H), 2.86-2.80 (m, 1H), 1.12-1.08 (m, 2H), 0.97-0.94 (m, 2H). 20 ”Example 17: Synthesis of (S)-4-ethyl-8-toco-4-hydroxy-9-methyl-11-(4-hydroxybutyl)amino-Lb12-di-14H-pyrano[3',4:6,7] 94] BRESFF [1,2-b] EMK-3,14(4H)-— Bd (STI-F6) sy AcO HO = N 17a NH ce) LiOH / MeOH NH ce) F AcO = © NNF AcO = 0 F HO' 20 9b 17b STI-F6 Step 1: Compounds 9b (91 mg, 0.20 mmol), 17a (52 mg, 0.40 mmol), Xantphos (23 mg, 0.04 mmol), ZARFE (5 mg, 0.02 mmol), and Fi ARIE (195 mg, 0.60 mmol) from Example 9 were added to a reaction flask and incubated for 5 hours at 100°C using a 6 mL TUK HRTEM, SRP. FRE. The system was concentrated to dryness and purified by silica gel column chromatography.(DCM:MeOH=100:1), #22!) 20 mg of compound 17b, a pale yellow solid, yield 18.2%. LCMS (254 nm) purity 97.1%, Rt = 1.89 min; MS calculated value: S$$1.2; MS measured value: 5$52.3 [M+HT]. IH NMR (400 MHz, DMSO-ds) 5 8.30 (d, J = 8.1 Hz, 1H), 7.56 (d, J= 11.1 Hz, 1H), 7.37 (t, J = 5.8 Hz, 1H), 6.87 (s, 1H), 5.46 (s, 2H), 5.50-5.35 (m, 2H), 4.11-4.03 (m, 2H), 3.67-3.61 (m, 2H), 2.43 (s, 3H), 2.20 (s, 30 3H), 2.20-2.03 (m, 2H), 2.00 (s, 3H), 1.75 (h, / = 4.3 Hz, 4H), 0.90 (t, J= 7.4 Hz, 3H). WR 2: Compound 17b (35 mg, 0.06 mmol HF) was added to 2 mL of methanol and 1 mL of KOMP RAIA, On 65 WO 2024 / 255740 PCT / CN2024 / 098491 A LiOH (35 mg, 1.46 mmol), AUER, img FE for 80 min. The pH was adjusted to 3-4 with glacial acetic acid, and the mixture was concentrated and prepared by reverse mixing under medium and low pressure (acidic mobile phase). The solution was lyophilized to give 15 mg of compound STI-F6 as a light yellow solid, with a yield of 50.6%. LCMS (254 nm) purity 98.6%, Rt = 1.59 min; MS calculated value: 467.2; MS measured value: 468.3 [M+H]*. 1H NMR (400 MHz, DMSO-ds) 8 8.43 (d, J = 7.7 Hz, 1H), 7.57 (d, J= 10.3 Hz, 1H), 7.45 (s, 1H), 5 6.61 (s, 1H), 5.46 (s, 2H), 5.44 (s, 2H), 3.74 (q, J = 6.7 Hz, 2H), 3.48 (t, J = 6.4 Hz, 2H), 2.43 (s, 3H), 1.96 — 1.73 (m,4H), 1.66 — 1.54 (m, 2H), 0.87 (t, J= 7.3 Hz, 3H). Intermediate compound 17a was prepared as follows: = Ac2O AcO TFAIDCM AcO NHBoc Pyridine a SN. 17a-1 17a-2 17a Step 1: Compound 17a-1 (3.0 g, 15.9 mmol ¥4F 15 mL HEE, DA 15 mL ZT, AUR 10 FP, 40°CAFARM LZ. US Bok AD EA 2EHY, A ALARAA washed with 2M hydrochloric acid to adjust pH < 2, then the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated by rotary evaporation to obtain 3.56 g of compound 17a-2 as a transparent oil, with a yield of 97.1%. 1H NMR (400 MHz, DMSO-d6) δ 6.81 (t, J= 5.8 Hz, 1H), 3.98 (t, J= 6.6 Hz, 2H), 2.92 (q, J= 6.6 Hz, 2H), 1.99 (s, 3H), 1.59 — 1.49 (m, 2H), 1.45 — 1.33 (m, 11H). Step 2: Compound 17a-2 (92 mg, 0.4 mmol) was dissolved in 0.5 mL of dichloromethane, 0.5 mL of HAAR was added, the reaction was carried out at room temperature for 20 minutes under 15" nitrogen protection. After rotary evaporation to dryness, crude compound 17a was obtained, which can be directly used in the next reaction. Example 18: Synthesis of (S)-11-((4-aminobutyl)amino)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-diaza-14H-pyrano[3',4':6,7]S| PRBS IF [1,2-b]FEMK-3,14(4H)-di one (STI-F7) BocHN BocHN HN cl fe) “CF H aeeete, 一 ion RN O 1) TFA / DCM a 了 F AcO 20 aerate, 2) LIOH / MeOH et F Aco = F How: _ 0 9b 18b STI-F7 20 Step 1: Compound 9b of Example 9 (91 mg, 0.2Xantphos (23 mg, 0.04 mmol), acetic acid (5 mg, 0.02 mmol), and tungsten carbonate (130 mg, 0.4 mmol) were added to a reaction flask under nitrogen protection. 18a (75 mg, 0.4 mmol) was added to 6 mL of anhydrous methyl methacrylate, stirred thoroughly, and heated to 100°C. The reaction was carried out for 1.0 h. SIN. TLC (DCM:MeOH = 30:1, product Rf = 0.3) was used to detect the reaction. After completion, the system was concentrated under reduced pressure and purified by column chromatography (DCM:MeOH = 100:1) to give 100 mg of compound 18b as a brown solid, yield 54.8%. ¹H NMR (400 MHz, 25 DMSO-ds) ¹H NMR (400 MHz, 25 DMSO-ds) ¹H NMR ¹⁸ ... J= 11.2 Hz, 1H), 7.40 (br, 1H), 6.87 (s, 1H), 6.83 (t, J =5.6 Hz, 1H), 5.46 (s, 2H), 5.53 (d, J =4.8 Hz, 2H), 3.60 (q, J =6.4 Hz, 2H), 2.97 (q, J =6.4 Hz, 2H), 2.44 (s, 3H), 2.20 (s, 3H), 2.12-2.07 (m, 2H), 1.70-1.63 (m, 2H), 1.56-1.49 (m, 2H), 1.33 (s, 9H), 0.90 (t, J = 7.2 Hz, 3H). Step 2: Compound 18b (100 mg, 0.16 mmol) was added to DCM (1.5 mD), followed by trifluoroacetic acid (0.5 mL). The reaction was carried out at room temperature for 20 minutes, then stopped. The system was concentrated to dryness under reduced pressure, and methanol (2.0 mD) and LiOH (100 mg) were added. The mixture was stirred at room temperature for another 20 minutes, then the reaction was stopped. The pH was adjusted to 4-5 with acetic acid. After concentration, the mixture was prepared by reverse top reaction under medium-low pressure (acidic mobile phase). The solution was lyophilized to 60 mg of compound STLF7, which was a pale yellow solid with a yield of 83.3%. LCMS (254 nm) 66 WO2024 / 255740 PCT / CN2024 / 098491 Purity 100.0%, Rt = 1.789 min; MS calcd.: 466.2; MS found: 467.2 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 8.39 (d, J = 8.0 Hz, 1H), 8.06 (br, 1H), 7.73 (br, 3H), 7.61 (d, J = 10.8 Hz, 1H), 7.36 (s, 1H), 6.56 (br, 1H), 5.46 (s, 2H), 5.43 (s, 2H), 3.60 (q, J = 6.4 Hz, 2H), 2.97 (q, J = 6.4 Hz, 2H), 2.44 (s, 3H), 1.91-1.81 (m, 2H), 1.79-1.67 (m, 4H), 0.87 (t, J = 7.2 Hz, 3H). 5 SCH 19: (S)-4-Z,28-8-Fl-4-FEHE-9-ABEL. 1 1-[RAESAIE-1,12- = S-14 H-A SF [3',4":6,7] 5] BR I+ [1,2-b] SOK-3,14(4H)-— A (STI-FI3) Synthesis Cl fe) 一 一 、Nh: 一 一 — N HN oO _ HN re) H₃C o~ / D 一 ~ = N _ HOH 一 N N H₃C o\ fo H₃C a ff np F AcO”= 0 N N — F AcO 20 F HO 20 9b 19a STI-F13 Step 1: Compound 9b (91 mg, 0.20 mmol), n-pentylamine (35 mg, 0.40 mmol), Xantphos 10 (23 mg, 0.04 mmol), Z.BR48 (5 mg, 0.02 mmol) and Al FRES HAC 30 mg (0.40 mmol) of Example 9 were added into a reaction flask, dissolved with 6 mL of anhydrous toluene, heated to 100°C under nitrogen protection, and reacted for 5 hours. The system was concentrated to dryness, and purified by silica gel column chromatography (DCM:MeOH = 200:1~100:1), 742] 30 mg of compound 19a was obtained as a light red solid, with a yield of 29.6%. Step 2: The compound19a (23 mg, 0.05 mmol) was mixed with 2 mL of methanol and 1 mL of dioxane, and A. LiOH (23 mg, 0.96 mmol) was added. Under nitrogen protection, the mixture was stirred at room temperature for 80 minutes. The pH was adjusted to 3-4 with glacial acetic acid, and the mixture was concentrated and prepared by reverse mixing at low pressure (acidic mobile phase) at 15" for lyophilization to obtain 12 mg of compound STI-F13 as a pale yellow solid, with a yield of 56.9%. LCMS (254 nm) purity 98.5%, Rt=1.97 min; MS calculated value: 465.2; MS measured value: 466.3 [M+H]*. 1H NMR (400 MHz, DMSO-de) 5 8.41 (d, J = 7.7 Hz, 1H), 7.55 (d J = 10.3 Hz, 1H), ...7.55 (d J = 10.3 Hz, 1H), 7.55 (d J = 10.3 Hz, 1H), 7.55 (d J = 10.3 1H), 7.42 (s, 1H), 6.60 (s, 1H), 5.50- 5.37 (m, 4H), 3.72-3.67 (m, 2H), 2.42 (s, 3H), 1.87 (dq, J = 14.1, 7.1 Hz, 2H), 1.74 (q, J= 7.4 Hz, 2H), 1.41 (td, J= 12.8, 7.1 Hz, 4H), 0.90 (dt, J= 18.2, 7.2 Hz, 6H). 20 Example 20: (S)-N-(2-(9-A FE -4-Z, HE-8-el -4-F HE-3,14- — AR -3,4,12,14- PU St -1 A ot By [3',4':6,7] "| Synthesis of PRE FF [1 ,2-b] EMK-1 1-2) Z,58)-NF ASE EH CSTI-G4) "Cor HzSO4, HNOg "Cr Fe, mace" "Cr" te HOME _ 9 OCR ea 20b 20¢ 04 Home HC , fe) . Machine ov of FeSQ4, H»SO4, ACOH, fBUOOH and people, IPrNHp, ¢-HCl shed? Cl ed Ho N DMSO "sy ON EtyN, DCM Br ~ IF How: oO Br aN fo "ae ° 20e ~ F How: Oo F HO' 20 20f a 20g / 1) BocNH2C82CO3,Pd(OAc)>,Xantphos = N 9 2) TFA, DCM HN vo' io F HO' Ro STI-G4 67 WO 2024 / 255740 PCT / CN2024 / 098491 Step 1: Compound 20a (2.0 g, 10.0 mmol) JA TAAR (.2 mL, 28.0 mmol) was reacted with sulfuric acid (8.0 mL) AVE in a combined solvent at room temperature for 2 hours. TLC (PE:EA=10:1, product R ≥ 0.7) confirmed the end of the reaction, poured the system into ice water, took a certain amount of ethyl acetate, dried the organic phase with anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained brown crude product was purified by column chromatography (PE:EA=50:1) to give 2.2 g of compound 20b, as a yellow solid, with a yield of 89.1%. 'H NMR (400 MHz, DMSO-ds) 5 810.1 (s, 1H), 8.32 (d, J = 8.4 Hz, 1H), 7.26 (d, J = 6.8 Hz, 1H). Step 2: Compound 20b (247 mg, 1.0 mmolD), iron powder (173 mg, 3.1 mmolD), and glacial acetic acid (1.0 mL, 18.0 mmol) were added to a mixture of ethanol (3.0 mD) and water (1.0 mD). The mixture was precipitated at 80 CF Bey 1 AS, TLC (PE:EA = 10:1, R0.5 for product) to confirm the reaction was complete. The system was allowed to cool naturally to room temperature, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (PE:EA = 50:1) to obtain 190 mg of compound 20c. It was a yellow solid, with a yield of 81.5%. ¹H NMR (400 MHz, 10 DMSOO-ds) 9.75 (s, 1H), 7.92 (d, J = 8.0 Hz, 1H), 7.44 (s, 2H), 6.68 (d J = 11.6 Hz, 1H). Step 3: Compound 20c (233 mg, 1.0 mmol) was FLAY 1c (121 mg, 1.0 mmol) HF ICA AA 5 mL, AUR. DIKES 30 Se. DATFASTA 1 mg, 0.06 mmOD, refluxed and stirred for 1 hour. After the reaction was complete as determined by LCMS, the reaction system was cooled to room temperature, concentrated to dryness under reduced pressure, and eluented with acetone to give 260 mg of compound 20d as a yellow solid, yield 8.6%. ¹H NMR (400 MHz, DMSO-ds) 8 8.69 (s, 1H), 8.66 15 (d, J=8.0 Hz, 1H), 8.12 (d J= 10.0 Hz, 1H), 7.35 (s, 1H), 6.53 (br, 1H), 5.43 (s, 2H), 5.29 (s, 2H), 1.92 - 1.81 (m, 2H), 0.88 (t, / = 6.4 Hz, 3H). Step 4: Compound 20d (222 mg, 0.5 mmol) and RGAE HBL FR (52 mg, 0.19 mmolD) were added sequentially to 8 mL of water, cooled to 0-5 °C, and then glacial acetic acid (0.52 mL, 9.2 mmolD) was added under nitrogen protection. Add sulfuric acid (3.6 mL), [Al] and add butanol peroxide (0.34 mL, 3.5 mmol) dropwise to the system, and react at room temperature for 3 hours. Pour the system into ice water, collect 20 μL, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and purify by silica gel column chromatography (DCM:MeOH = 100:1) to give 130 mg of compound 20e, as a trademark-colored solid, with a yield of 56.8%. ¹H NMR (400 MHz, DMSO-do) 5 8.65 (d, J = 7.6 Hz, 1H), 8.10 (d J = 10.0 Hz, 1H), 7.33 (s, 1H), 6.53 (s, 1H), 5.43 (s, 2H), 5.29 (s, 2H), 2.79 (s, 3H), 1.91 - 1.81 (m, 2H), 0.88 (t, J = 7.6 Hz, 3H). Step 5: Add compound 20e (92 mg, 0.2 mmol / L) to 2 mL DMSO, and under nitrogen protection, add hydrochloric acid (0.12 mL, 1.4 mmol / L) to the reaction system at 25 °C.Isopropylamine (0.1 mL, 1.2 mmol), F 140 AY 1 hour. After LCMS analysis, the system was concentrated under reduced pressure to a certain level, and then prepared under medium-high pressure in reverse (acid mobile phase). The resulting product was lyophilized to obtain 35 mg of compound 20f as a yellow solid, with a yield of 33.0%. Step 6: Compound 20f (30 mg, 0.057 mmol) was added to 2 mL of DCM, cooled to 0-20°C, and methyl thiocyanate (8 mg, 0.068 mmol) and triethylamine (7 mg, 0.068 mmol) were added to the reaction system. The mixture was then fermented for 30 hours. After LCMS analysis, the organic layer was washed sequentially with 1M hydrochloric acid aqueous solution and saturated brine. The organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain 30 mg of crude compound 20g, which was directly used in the next step. Step 7: The crude compound 20g (30 mg, 0.049 mmol) and tert-butyl carbamate (12 mg, 0.049 mmol) were added to the reaction system. Dissolve 3 mL of anhydrous methane (33 mg, 0.01 mmol) in nitrogen-protected solution, add carbonate (33 mg, 0.01 mmol), Xantphos (1 mg, 0.001 mmol), All Z 440.2 mg, 0.001 mmol), HPN, FHA 100°C, and react for 2.0 h. After the reaction is complete as detected by LCMS, concentrate the system to dryness under reduced pressure, remove dioxane and water, concentrate the organic phase, add 2 mL of 25% trioxane-acetic acid in dioxane solution to the residue, stir at room temperature for 30 h, and after the reaction is complete as detected by LCMS, concentrate the system to dryness under reduced pressure and prepare by reverse high pressure (acid mobile phase), freeze-dry to obtain 8 mg of compound STI-G4 as a yellow solid, yield 30.0%. 68 WO 2024 / 255740 PCT / CN2024 / 098491 Similarly, the following compound from the examples was synthesized: 21 HO an O 〇= NF HO 29 STI-F12R 22 HOa = NF HO 20 STI-F10 Example P1:, (S)-4-ethyl-8-amino-4-hydroxy-11-((2-carboxyethyl)amino)-9-methyl-1,12-dioxo-14H-pyridine[3,,4':6,7] 1] BE [1,2-b] ZEWK-3,14(4H)-— A CSTI-FS) Synthesis esotb -s-02b on aco” F Aco”: 0 F How: 5 F5-03b F5-04b STI-F5 Step 1: Dissolve compound F5-01b (1.22 g, 20.00 mmol) = Z. / K (4.17 mL, 30.00 mmolD) in 20 mL TA, AEA T Se reed = $e A RG (6.34 g, 24.00 mmol) AY 10 mL = A bey solution, added dropwise over 3 minutes, under nitrogen protection, reacted at room temperature for 1.5 hours. Added 30 mL KAR, A eR, BSF 10" organic phase, dried over anhydrous sodium sulfate, and the solvent was rotary evaporated to give 3.5 g of compound F5-02b as a colorless oil, yield 100%. ¹H NMR (400 MHz, DMSO-ds) 3.57 (t, J = 5.8 Hz, 2H), 2.67 (t, J = 5.8 Hz, 2H), 0.87 (s, 9H), 0.04 (s, 6H). Step 2: 3%(S)-11-Sl-4-Z.5k-8-F-9-FE-3,14-— A 3,4, 12, 14- VU Si - 1 At [3',47:6, 7] 5] [1,2-b] ZS U-4-38 ZA (184 mg, 0.40 mmol). F5-02b (140 mg, 0.80 mmol), Xantphos (23 mg, 15 0.04 mmol), ZARFEO mg, 0.04 mmol, and carbonated beverage (391 mg, 0.80 mmol) JIA BIH, FA 10 mL anhydrous formaldehyde was dissolved, and under nitrogen protection, the mixture was heated to 100°C and reacted for 1.5 hours. The system was concentrated to dryness and purified by silica gel column chromatography (DCM:MeOH = 200:1) to give 127 mg of compound F5-03b, which was a light-coloredYellow solid, yield 52.9%. ¹H NMR (400 MHz, DMSO-ds) ¹H NMR: 8.23 ​​(d, J = 8.2 Hz, 1H), 7.56 (d, J = 11.1 Hz, 1H), 7.52 (t, J = 6.5 Hz, 1H), 6.86 (s, 1H), 5.46 (s, 2H), 5.43 (s, 2H), 3.83 (t, J = 5.6 Hz, 2H), 3.70 (t, J = 5.5 Hz, 2H), 2.43 (s, 3H), 2.20 (s, 3H), 2.16–2.08 (m, 2H), 0.89 (t, J = 7.6 Hz, 3H), 0.67 (s, 9H), -0.11 (s, 3H), -0.13 (s, 3H). Step 3: Add F5-03b (127 mg, 0.21 mmol JAF 1 mL DAH KE, A 4M HCl AY 14- Dioxane 69 WO 2024 / 255740 PCT / CN2024 / 098491 TAR, AER, SEE) to 12 mL formaldehyde, add lithium oxide (150 mg), AAR, and stir at room temperature for 1 hour. Adjust the pH to 6-7 with acetic acid, prepare by high pressure reverse reaction (mobile phase: acetonitrile / 0.05% formic acid aqueous solution), freeze for 5 minutes to obtain 16 mg. Compound STFS, a pale yellow solid, yield 56.9%. LCMS (254 nm, purity 99.7%, Rt = 1.45 min; MS calculated value: 439.2; MS measured value: 440.1 [M+H-]. IH NMR (400 MHz, DMSO-ds) 5 8.27 (d J = 8.1 Hz, 1H), 7.57 (d, J = 11.1 Hz, 1H), 7.37 (t, J = 6.1 Hz, 1H), 7.20 (s, 1H), 6.46 (s, 1H), 5.43 (s, 2H), 5.40 (s, 2H), 5.00 (s, 1H), 3.69 (d, J = 2.9 Hz, 4H).2.43 (s, 3H), 1.94 — 1.76 (m, J = 7.2 Hz, 2H), 0.87 (t, J= 7.3 Hz, 3H). 10 Example P2: Synthesis of (S,E)-4- Z.2k-8-Fl-4-FEE- 1 1-((4-FESE-2-M- 1-2) AH) -9- FA E-1,12-— Sh -1 4-H [3',4':6,7] §|GE[1,2-b] EUK-3,14(4H)-— Ae] CSTLF8) Oo Ho Coe O O HON OH 5 日 OTBS F8-01 F8-02 F8-02c F8-01b TBSO HO HO a Oo a Oo a ce) 一 > = N 站 = N — = N F Aco”2 “0 F AcO™= 0 F Ho" = _O F8-02b F8-03b STI-F8 Step 1: Compound F8-01 (1.2 g, 13.60 mmol) was dissolved in 40 mL PUA, cooled to 0°C, under nitrogen protection, 15 add phthalimide (1.00 g, 6.80 mmol), then add diethyl azodicarboxylate (3.$7 g, 13.60 mmol) in 60 mL TAY, and react at room temperature for 2 hours. The solvent was rotated to evaporate, purified by silica gel column PE:EA=5$:1D) to obtain 500 mg of compound F8-02 as a white solid. !IH NMR (400 MHz, DMSO-ds) δ 7.93 — 7.80 (m, 4H), 5.74 — 5.60 (m, 2H), 4.71 (t, J = 5.5 Hz, 1H), 4.21 — 4.15 (m, 2H), 3.90 (m, 2H). Step 2: F8-02 (759 mg, 3.5 mmol), = Z.f¥(1062 mg, 10.5 mmol) was dissolved in 8 mL dichloromethane, 20 tert-butyldimethylsilyl chloride (633 mg, 4.2 mmol) in 2 mL dichloromethane solution was slowly added dropwise, under nitrogen protection, stirred at room temperature FER. WEAR, TEBE (PE:EA=10:1), 4421] 536 mg of compound F8-02c was obtained as a colorless oil. 'H NMR (400 MHz,DMSO-ds) 8 7.93 — 7.80 (m, 4H), 5.77 — 5.61 (m, 2H), 4.21 — 4.15 (m, 2H), 4.10 (dd, J = 2.9, 1.5 Hz, 2H), 0.83 (s, 9H), 0.00 (s, 6H). Step 3: Add 8 mL of anhydrous ethanol to 80% 7K AHF (112 mg, 1.78 25 mmol), FRM 6 ZN. SOE, weer. MANS, FE, eer. to 250 mg of compound F8-0l1b, which is a yellow oily substance. Step 4, (S)-11-oxo-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetraaza-1H-pyran [3.4'2:6,7] tincture 70 WO 2024 / 255740 PCT / CN2024 / 098491 [1,2-b] Ze UR-4-3 Z, RH (184 mg, 0.40 mmol), Xantphos (23 mg, 0.04 mmol), Acetic acid (9 mg, 0.04 mmol), FPR ASH (391 mg, 0.80 mmol), JA RMI, JA F8-01b (161 mg, 0.80 mmol), I 10 mL TOK PAAR, AUER. FRR 100°C DY 1 hour. Concentrate the system to dryness. Purification by silica gel column chromatography (DCM:MeOH = 200:1~150:1), #42] 100 mg of compound F8-02b, as a standard solid, yield 39.9%. MS tt 5 SHE: 621.2; MS measured value: 622.3 [M+H]*. Step $: Mix 3 mL of a solution of F8-02b (100 mg, 0.16 mmol) in dioxane and 4 M HCl with 1,4-dioxane (SmD), under nitrogen protection, stir at room temperature for 1 hour, evaporate to dryness, and use the crude product directly for the next step. LCMS (254 nm) purity 85.0%, Rt = 1.67 min; MS calculated value: 507.2; MS measured value: 3508.2 [M+H] . Step 6:The crude product obtained in the previous step was dissolved in 8 mL of He, OA, 38 mg ASU, BU SU BY 1 10 MT. VA RAK pH to $, and prepared under high pressure in reverse (mobile phase: acetic acid / 0.05% formic acid aqueous solution). After freezing, 35 mg of compound STIL-F8 was obtained as a pale yellow solid. LCMS (254 nm, purity 97.5%, Rt = 1.50 min; MS calculated value: 465.2; MS measured value: 466.2 [M+H]). ¹H NMR (400 MHz, DMSO-ds) 8.34 (d, J = 8.1 Hz, 1H), 7.77 (t, J = 6.3 Hz, 1H), 7.61 (d, J = 11.1 Hz, 1H), 7.25 (s, 1H), 6.51 (s, 1H), 5.94 (dt, J = 15.8, 4.5 Hz, 1H), 5.71 (dt, J = 15.7, 5.1 Hz, 1H), 5.45 (s, 2H), 5.37 (s, 2H). 4.75 (s, 1H), 4.28 (t, J= 5.8 Hz, 15 2H), 3.97 (d, J= 4.9 Hz, 2H), 2.47 (s, 3H), 1.97 — 1.85 (m, 2H), 0.93 (t, J= 7.3 Hz, 3H). Example P3: (S)-4- 2 F&-8-H -4-F2 HA-1 1-(4-(FE AE) ORE -1-3E)-9- FR BE -1,12-—= Sh -1 4 OB [3',4°:6,7] 3] PR[1,2-b] EUK-3,14(4)-— A] CSTI-F14) Synthesis of OAc OH OH OAc OAc ia a BS - & - & - oo on, F14-01 F14-02 F14-03 / Aeon / Ho F14-04 STI-F14 20 Step 1: Add compound F14-01 (1.00 g, 4.60 mmol) to 20.0 mL of dioxane, add pyridine (790 mg, 9.20 mmol) #ll ZH (630 mg, 7.40 mmol), under nitrogen protection, and react overnight at room temperature. Add 1M hydrochloric acid aqueous solution under ice bath to adjust pH to 1-2, then add ethyl acetate.The ester was washed with saturated sodium ammonium carbonate, then with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was vortexed and dried to give 1143 mg of compound F14-02 as a pale yellow oil, yield 96.6%. ¹H NMR (400 MHz, DMSO-ds) 5 3.94 (d, J = 13.0 Hz, 2H), 3.86 (d, J = 6.5 Hz, 2H), 2.69 (s, 2H), 2.01 (s, 3H), 1.81 — 25 1.71 (m, 1H), 1.61 (dd, J = 12.7, 3.5 Hz, 2H), 1.39 (s, 9H), 1.12 — 0.97 (m, 2H). GRR 2: 1 F14-02 (680 mg, 2.64 mmol AF) was added to 5 mL of dioxane, followed by 3 mL of trifluoroacetic acid and a protective atmosphere. The mixture was stirred at room temperature for 1 hour, and the reaction solution was evaporated to dryness. Product F14-03 was used directly for the next step of the reaction. Step 3: (S)-11-oxo-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetraoxo-1H-pyridine[3,4'2:6,7]oxo[12-b]azolin-4-yl acetate (184 mg, 0.40 mmol OD), Xantphos (23 mg, 0.04 mmol OD), acetate (9 mg, 0.04 mmol OD), and levonorgestrel (391 mg, 0.80 mmol OD) were added to the reaction flask and mixed with 10 mL of dioxane. Anhydrous methyl methacrylate was hydrolyzed, and F14-03 (217 mg, 0.80 mmol) was added to 1 mL AT BEAK, FUP. The reaction was carried out at 100°C for 1 hour. The system was concentrated to dryness and purified by silica gel column chromatography (DCM:MeOH = 300:1~100:D) to give 120 mg of compound F14-04 as a pale yellow solid, yield 52.0%. LCMS (254 nm, purity 82.5%, Rt = 2.04 min; MS calculated value: 577.2; MS measured value: 378.3 [M+H]). 71 WO 2024 / 255740 PCT / CN2024 / 098491 Step 4:F14-04 (80 mg, 0.14 mmol HAF, 5 mL methanol) was added to lithium oxyoxide (33 mg, 1.40 mmol), and stirred at room temperature for 40 minutes under nitrogen protection. The pH was adjusted to ~6 with glacial acetic acid, and GSA (ZI / 0.05% fluoroacetic acid aqueous solution) was prepared by reverse high-pressure chromatography. The mixture was lyophilized to give 15 mg of compound STLF14 as a pale yellow solid, yield 21.7%. LCMS (254 nm) purity 99.3%, Rt = 1.67 min; MS calculated value: 493.2; MS measured value: 494.3 [M+H]*. ¹H NMR (400 MHz, 5 DMSO-ds) 7.91 (d, J = 8.4 Hz, 1H), 7.74 (d, J = 10.8 Hz, 1H), 7.29 (s, 1H), 6.58 (d, J = 63.2 Hz, 2H), 5.46 (s, 2H), 5.42 (s, 2H), 3.69 — 3.64 (m, 2H), 3.42 (d, J = 6.2 Hz, 2H), 3.25 (t, J = 11.5 Hz, 2H), 2.47 (s, 3H), 1.93 — 1.78 (m, 4H), 1.74 — 1.65 (m, 1H), 1.58 — 1.45 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). 10 “Example P4: (S)-4-ethyl-8-amino-4-hydroxy-1L-((lr,4S)-4-(hydroxymethyl) Synthesis steps of compound F15-01 (1.50 g, 11.62 mmol D, ditert-butyl dicarbonate (3.04 g, 13.94 mmol AF)) were performed overnight at room temperature under nitrogen protection. The solvent was rotary evaporated, and the product was washed with saturated brine, dried over 15° anhydrous sodium sulfate, filtered, and the filtrate was rotary evaporated to dryness to give 2.74 g of compound F15-02, which was a white solid.100% efficiency. ¹H NMR (400 MHz, DMSO-ds) 6.66 (d, J = 8.0 Hz, 1H), 4.36 (t, J = 0.3 Hz, 1H), 3.18 (t, J = 5.8 Hz, 2H), 3.15 — 3.06 (m, 1H), 1.80 — 1.67 (m, 4H), 1.37 (s, 9H), 1.29 — 1.18 (m, 1H), 1.17 — 1.02 (m, 2H), 0.94 — 0.80 (m, 2H). Step 2: Compound F15-02 (1.10 g, 4.80 mmol) was added to 2.5 mL of pyridine, protected with 20” AR, A 20”, and heated to 40 sC. The reaction was heated for 4 hours. 1M hydrochloric acid aqueous solution was added under ice bath to adjust the pH to 1-2. Ethyl acetate was added, saturated brine was added, and the mixture was dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was evaporated to dryness to give 979 mg of compound F15-03 as a white solid, yield 75%. Step 3: F15-03 (740 mg, 2.73 mmol WF) was added to 5 mL of dichloromethane and 3 mL of trifluoroacetic acid. Under nitrogen protection, the mixture was stirred at room temperature for 2 hours. The reaction mixture was evaporated to dryness and used directly for the next step. ¹H NMR (400 MHz, 25 DMSO-ds) ¹³⁸ 3.83 (d, J = 6.4 Hz, 2H), 3.01–2.87 (m, 1H), 2.01 (s, 3H), 1.98–1.89 (m, 2H). 2H), 1.80 — 1.71 (m, 2H), 1.60 — 1.47 (m, 1H), 1.35 — 1.21 (m, 2H), 1.11 —0.96 (m, 2H). Step 4: (S)-11-oxo-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetraoxo-1H-pyrrolidine[3.4'2:6,7]oxosin[12-b]azolin-4-yl acetate (184 mg, 0.40 mmol), Xantphos (23 mg, 0.04 mmol), acetate (9 mg, 0.04 mmol), fi RARE (391 mg, 0.80 mmol) JIASVR was dissolved in 10 mL of anhydrous methyl methacrylate, and 1 mL of A) RAK of FE15-04 (114 30 mg, 0.80 mmol) was added. Under nitrogen protection, the mixture was heated to 100°C and reacted for 1.5 hours. The system was concentrated to dryness and purified by silica gel column chromatography (DCM:MeOH = 300:1~100:1). 135 mg of compound F15-05 was obtained as a pale yellow solid, with a yield of 57.1%. LCMS (254 nm) purity 79.8%, Rt = 1.97 min; MS calculated value: S91.2; MS measured value: 392.3 [M-+HH]. Procedure: F15-05 (135 mg, 0.23 mmol) was added to 10 mL of methanol, and lithium oxyoxide (55 mg, 2.30 mmol) was added. Under nitrogen protection at 35°C, the mixture was stirred at room temperature for 1 hour. The pH was adjusted to 4- with glacial acetic acid. GRBYAH was prepared by reverse high pressure: ZR / 0.05%= 72 WO 2024 / 255740 PCT / CN2024 / 098491 acetic acid aqueous solution), and lyophilized to obtain 10 mg of compound STLFIS, AYRE IA, £728 8.6%. LCMS (254 nm) purity 95.8%, Rt = 1.62 min; MS calculated value: S91.2; MS measured value: 392.3 [M-+HH]. Calculated value: 507.2; MS measured value: 308.3 [M+HH]. IH NMR (400 MHz, DMSO-ds) 8 8.50 (d, J= 7.9 Hz, 1H), 7.59 (d, J= 10.5 Hz, 1H), 7.41 (s, 1H), 6.56 (s, 1H), 5.43 (s, 2H), 5.40 (s, 2H), 3.77 (s, 1H), 3.31 (d, J = 6.0 Hz, 2H), 2.45 (s, 3H), 2.11 — 1.97 (m, 2H), 1.94 — 5 1.79 (m, 4H), 1.59 (q, J= 11.8 Hz, 2H), 1.42 (s, 1H), 1.22 — 1.10 (m, 2H), 0.87 (t, J = 7.3 Hz, 3H). Example PS: (S)-4-ethyl-8-fluoro-4-hydroxy-11-((lr, 4S)-4-(hydroxymethyl)cyclohexyl)Synthesis of N-9-methyl-1,12-dioxo-14H-HbMj [3',4':6,7] 1 9e[1,2-b] SOK-3,14(4H)-— Be] (CSTLEF17) Step 1: Compound F17-01 (1.00 g, 4.92 mmol) was dissolved in 5 mL AGE, DLA 5 mL ZR, BER and heated at 40°C for 4 hours. Add 4M hydrochloric acid aqueous solution under bath pressure to adjust pH to 1-2, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, and evaporate the filtrate to dryness to obtain 1.12 g of compound F17-02 as a transparent oil, yield 92.9%. ¹H NMR (400 MHz, CDCls) 5 4.56 (s, 1H), 4.06 (J = 6.6 Hz, 2H), 3.12 (q, J = 6.7 Hz, 2H), 2.05 (s, 3H), 1.64 (h, J = 6.2 Hz, 2H), 1.55 — 1.49 (m, 2H), 1.45 (s, 9H), 1.42 — 1.34 (m, 15 2H). Step 2: Add F17-02 (1.12 g, 4.57 mmol JAF 9 mL MARK, 1AS mL triacetic acid, nitrogen protection, stirred at room temperature for 3 hours, vortex and dry the reaction solution, crude product F17-03 directly used for the next step of the reaction. Step 3: Dissolve (S)-11-m-4-ethyl-8-fluoro-9-methyl-3,14-dioxo-3,4,12,14-tetram-1-pyridine[3.4':6,7] oxo[1,2-b]MSMt-4-3E 7, WHE (184 mg, 0.40 mmol). Xantphos (46 mg, 0.08 mmol), methyl acetate (9 mg, 0.04 mmol), PAR HA (S21 mg, 1.60 mmol), JA HAP, in 10 mL anhydrous methyl acetate, WHE (184 mg, 0.40 mmol), Xantphos (46 mg, 0.08 mmol), methyl acetate (9 mg, 0.04 mmol), PAR HA (S21 mg, 1.60 mmol), JA HAP, in 10 mL anhydrous methyl acetate, Add F17-03 (114 mg, 0.80 mg)mmol) ff) 2 mL 1,4-—A7\ IA, AE. FP 100°C KADY. 100 minutes. The system was concentrated to dryness and purified by column chromatography (DCM:MeOH = 300:1~200:1). 300 mg of crude compound F17-04 was obtained. Step 4: F17-04 (140 mg, 0.25 mmol) was dissolved in 11 mL methanol, and lithium hydroxide (70 mg, 2.92 mmol) was added. The mixture was stirred at room temperature for 1 hour under nitrogen protection at 25°C. The pH was adjusted to 6 with glacial acetic acid, and the mixture was prepared by reverse high-pressure chromatography (mobile phase: methanol / 0.05% formic acid aqueous solution). The solution was lyophilized to obtain 15 mg of compound STILF17 as a pale yellow solid, with a yield of 12.5%. LCMS (254 nm) purity 99.7%, Rt = 1.55 min; MS calculated value: 481.2; MS measured value: 482.1. [M+H]'. 'H NMR (400 MHz, DMSO-ds) 5 8.26 (d, J = 8.1 Hz, 1H), 7.54 (d, J= 11.1 Hz, 1H), 7.31 (t, J = 5.9 Hz, 1H), 7.20 (s, 1H), 6.47 (s, 1H), 5.40 (s, 2H), 5.36 (s, 2H), 4.43 (t, J=5.5 Hz, 1H), 3.60 (q, J= 6.7 Hz, 4H), 2.42 30 (s, 3H), 1.93 — 1.76 (m, J= 7.2 Hz, 2H), 1.94 — 1.79 (m, 4H), 1.56 — 1.39 (m, 4H), 0.87 (t, J = 7.3 Hz, 3H). Example Po: Synthesis of (S)-7-ethyl-7-hydroxy-14-(4-hydroxybutyl)azono)-10,13-dioxo-11H-[1L3]dioxo[4.Sg]pyran[3',4':6,7]3]Gel 1,2-b]EMK-8,11(7H)-— A CSTI-F18) 73 WO 2024 / 255740 PCT / CN2024 / 098491 4? a bb < ALN < AN HE 1: Compound STI-E-05 (1.0 g, 2.66 mL of JHA (mmol) pyridine was added, followed by 6 mL of CET. Under A gas protection, the mixture was heated and stirred at 40°C, and monitored by LCMS. After the reaction was complete, the solution was concentrated under reduced pressure and slurried with acetone to obtain 360 mg of compound F9-01 as a yellow solid, with a yield of 33.0%. Step 2: Compound F9-01 (360 mg, 0.83 mmObD) was dissolved in 12 mL of acetic acid, and 4 mL of 30% peroxide aqueous solution was added. Under vacuum protection, the mixture was heated and stirred at 70°C for 3.0 hours. The reaction was monitored by LCMS until complete. The solution was concentrated under reduced pressure, and the residue was added to ice water. DCM was used to extract the residue, which was then dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure. 16 mL of DMF was added to the residue to dissolve it. 1.1 mL of oxaloyl chloride was added to the ice water, and the mixture was stirred at room temperature after the addition was complete. The reaction was monitored by LCMS. After the reaction was complete, water was added to extinguish the reaction, and the mixture was filtered. The filter cake was washed with deionized water and dried under reduced pressure to obtain 300 mg of compound F9-02 as a yellow solid. 77.3%. LCMS (254 nm, purity 92.13%, Rt = 1.83 min; MS calculated value: 468.1; MS measured value: 469.1 [M+H]*. Step 3: Compounds F9-02 (140 mg, 0.30 mmol), F18-S2 (122 mg, 0.60 mmol), Xantphos (35 mg, 0.06 mmol), Z.f4l! (7 mg, 0.03 mmol), FUTKEHA (392 mg, 0.60 mmol), JIA RVI, A 6 mL anhydrous methyl methacrylate and 6 mL 1, 4-H, BUR, Fr 100°C 52 2 h. The system was concentrated to dryness and purified by silica gel column chromatography (DCM:MeOH = 100:1) to obtain 50 mg Compound F18-01 is a red solid with a yield of 26.2%. LCMS (254 nm) showed a purity of 78.92%, Rt = 2.22 min; MS calculated value: 635.3; MS measured value: 636.3 [M+H]". Step 4: [M+H] of the compound...F18-01 (50 mg, 0.08 mmol) was reacted with 1 mL of dioxane hydrochloride bath solution containing MP bE and DA 4M at room temperature for 1 hour. After the reaction was completed, the system was concentrated to dryness under reduced pressure. 3 mL of methanol and lithium hydroxide (530 mg, 2.08 mmol / D) were added to the residue, and the mixture was stirred at room temperature for 20 minutes. The pH was adjusted to 3-4 with glacial acetic acid, and the mixture was concentrated and then prepared under high pressure in reverse (mobile phase: ZH5 / 0.05% = ZIRT). The solution was lyophilized to obtain 5 mg of compound STI-F18, a pale yellow solid with a yield of 13.3%. LCMS (254 nm) showed a purity of 97.86%, Rt = 1.38 min; MS calculated value: 479.2; MS measured value: 480.2 [M+H]. ¹H NMR (400 MHz, DMSO-ds) 7.94 (s, 1H), 7.67 (brs, 1H), 7.47 (s, 1H), 7.29 (s, 1H), 6.63 (d, J = 13.0 Hz, 2H), 6.31 (d, 1H), 5.46 (s, 2H), 5.43 (s, 2H), 3.70 (q, J = 7.0 Hz, 2H), 3.48 (d, J = 6.7 Hz, 2H), 1.92 — 1.82 (m, 2H), 1.81 — 1.71 (m, 2H), 1.61 — 1.54 (m, 2H), 0.88 (t, 25 J = 7.3 Hz, 3H). The intermediate compound F18-S2 was prepared as follows: HO TBSO TBSO ae AN. F18-S1 F18-S2 Step 1: Dissolve 4-(N-AUTAUS)-1-TBE (3785 mg, 20.0 mmol) in 20 mL of triethylamine (4.17 mL, 30.0 mmol), AT BAL Te = SE SUE GE (3617 mg, 24.00 mmol) 30 (N10 mL Ma A, SR, DUA AK AER, SARE, ASPABL) phase, dry with anhydrous sodium sulfate, and rotary evaporate the solvent to give 4.49 g of compound F18-S1 as a colorless oil, yield 74.1%. 74WO 2024 / 255740 PCT / CN2024 / 098491 'H NMR (400 MHz, DMSO-ds) 8 6.74 (t, J = 5.7 Hz, 1H), 3.54 (t, J = 5.6 Hz, 2H), 2.88 (q, J = 6.2 Hz, 2H), 1.40 — 1.38 (m, 4H), 1.35 (s, 9H), 0.87 (s, 9H), 0.04 (s, 6H). Step 2: Compound F18-S$1 (712 mg, 2.35 mmol WAT 5 mL dioxane, RP Ret Be O~S°C) was added, followed by trimethyliodosilane (604 mg, 2.82 mmol), and the reaction was carried out at a constant temperature for 3 minutes. Hours. TLC (DCM:MeOH = 10:1, product Rf = 0.2) detected the reaction after completion. Methanol (0.1 mID) was added to the reaction system, concentrated to dryness under reduced pressure, and purified by column chromatography (DCM:MeOH = 50:1). #2) 400 mg compound F18-S2, Wikis GAA, yield 83.8%. !IH NMR (400 MHz, DMSO-ds) 8 7.57 (s, 2H), 3.55 (t, J = 5.9 Hz, 2H), 2.76 (t, J = 7.5 Hz, 2H), 1.62 — 1.39 (m, 4H), 0.87 (s, 9H), 0.04 (s, 6H). 10 “Example P7: (S)-7- 2, 5k-7-FE-14-(4-FEET HE) BM HE)-10,13-= A-11 H-[1,3] =A [4,5-g] [3',4':6,7] 9] YE[1,2-b] -EMK-8,11(7H)-— A CSTI-GS) Synthesis an yen = ee plus nf Step 1: Compound G5-01 (1.4 g, 4.32 mmol) was added to 40 mL of ethanol / water (3:1D), along with iron powder (965 mg, 17.28 mmol / D) and tungsten oxide (231 mg, 4.32 mmol / D). The reaction mixture was reacted at 80°C for 15 hours. TLC monitoring showed complete conversion of the starting material. The mixture was filtered while hot, and the reaction solution was evaporated to dryness. The crude product was then subjected to column chromatography.Chromatographic purification yielded 1.0 g of compound GS-02, which was a colorimetric oil with a yield of 71.4%. LCMS (254 nm), Rt = 2.31 min; MS calculated value: 324.2; MS measured value: 323.2 [MH]. Step 2: Compound G5-02 (1.0 g, 3.09 mmol), 4-2 FA SEULNE (38 mg, 0.309 mmol), ¥ TF 30 mL NN-dimethylformamide, after replacing nitrogen, NN-diisopropylethylamine (1.2 g, 9.25 mmol), ZEAE Z 20” acyl chloride (842 mg, 6.18 mmol), FAA FRY 2 h. TLC monitoring showed complete conversion of the starting materials. Ethyl acetate and water were added to the reaction solution to decompose the reaction. After the reaction was carried out, 3 samples were taken, the organic layers were combined, washed with saturated brine and dried with anhydrous sodium sulfate. Wer. Hana 15 mL methanol, 1.0 g lithium hydroxide was added and the reaction was carried out at room temperature for 1 h. Hours. TLC monitoring showed complete conversion of the raw materials. Ethyl acetate and water were added to the reaction solution to quench the reaction, and the mixture was taken three times. The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain 750 mg of compound GS-03, a deep yellow oil, with a yield of 63.4%. G5-03 25 " The crude product was used directly in the next step. LCMS (254 nm), Rt = 2.21 min; MS calculated value: 382.2; MS measured value: 381.2 [MH]. Step 3: GS-03 (750 mg, 1.96 mmol HAF 10 mL 50%= AZARAE AIK, ZT fF) was precipitated for 1 hour, TLC WR SoA. LR DRACER IS AER 3 times, the organic layers were combined, washed with saturated brine and dried with anhydrous sodium sulfate, and then vortexed to dryness. The crude product was purified by column chromatography to give 450 mg of compound 30 "G5-04", which was a black oily substance with a yield of 81.4%. LCMS (254 nm), Rt = 1.70 min; MS calculated value:282.1; MS measured value: 283.3 [M+H] Step 4: Add (S)-4-ethyl-4-hydroxy-7,8-diA-1H-NMR [3,4-£]"5]PR-3,6,10(4H)-Al] (461 mg, 1.75 mmonD, and nitrogen protection to G5-04 (450 mg, 1.60 mmol JAF) and stir at 130°C for 30 min. Then add p-toluenesulfonic acid (69 mg, 0.40 mmonD) and continue to react at 130°C for 2 hours. During the reaction, a solid precipitated. After the reaction was completed, the reaction solution was evaporated and the crude product was slurried with acetone to obtain 520 mg of compound STI-G5, which was a gray solid. 75 WO 2024 / 255740 PCT / CN2024 / 098491 Yield 63.8%. LCMS (254 nm), Rt = 1.73 min; MS calculated value: 509.2; MS SEWI{A: 510.2 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 8 9.73 — 9.64 (m, 1H), 8.95 (d, J = 8.5 Hz, 1H), 8.01 (d, J = 12.0 Hz, 1H), 7.29 (s, 1H), 6.51 (s, 1H), 5.98 (s, 1H), 5.43 (s, 2H), 5.27 (s, 2H), 4.15 (s, 2H), 3.11 (d, J = 8.1 Hz, 2H), 1.95 — 1.79 (m, 2H), 1.73 (t, J = 7.8 Hz, 2H), 1.48 — 1.34 (m, 4H), 0.88 (d, J = 6.7 Hz, 5 6H). Synthesis of drug linker conjugate Example 23; Synthesis of drug linker conjugate STI-A3 2) 23d, EtaN, DMF p 10 Step 1, add compound STI-A (60 mg, 0.13 mmol), Boc-Glycine (46 mg, 0.26 mmol), and HATU (99 mg, 0.26 mmolD) from Example 3 to the reaction flask, purge three times with nitrogen, and add 1.5 mLDissolved in DMF, triethylamine (72 hL, 0.13 mmol / L, stirred at room temperature for 3.5 h). LCMS monitoring showed approximately 30% conversion, at which point the forward reaction was stopped. The product was prepared under medium-high pressure reverse reaction (alkali mobile phase), lyophilized to obtain 9 mg of compound 23a as a yellow solid. 45 mg of the starting material was recovered, yielding a yield of 53.6%. LCMS (254 nm) purity 98.32%, Rt = 1.98 min; MS calculated value: 636.3; MS SCHUH: 635.4 [MH]. ¹⁵ NMR (400 MHz, DMSO-ds) 8.23 ​​(d, J = 8.0 Hz, 1H), 7.83 (d, J = 10.6 Hz, 1H), 7.36 (s, 1H), 5.54 — 4.92 (m, 6H), 3.64 (d, 2H), 3.55 (t, J = 13.2 Hz, 2H), 2.51 (s, 3H), 1.87 (qt, J = 13.9, 6.8 Hz, 2H), 1.21 (s, 9H), 1.06 (s, 9H), 0.89 (t, J = 7.3 Hz, 3H). Step 2: Compound 23a (6 mg, 0.01 mmol) was added to 1 mL DCM, 0.3 mL TFA, and RARE F PELE for 0.5 hours. The reaction was monitored by LCMS until completion. The reaction was stopped, and the triethylamine (14 pL, 0.10 mmol), compound 23b (4 mg, 0.01 mmol), and HATU (8 mg, 0.02 mmol) were concentrated under reduced pressure. The reaction was purged with RH and nitrogen three times, dissolved in 1.0 mL of DMF, and stirred at room temperature for 1 hour. The reaction was confirmed by LCMS. The reaction was then prepared by high-pressure reverse reaction (alkali mobile phase), yielding compound 23c as a pale yellow solid (3 mg, yield 30.0%). LCMS (254 nm, purity 94.25%, Rt = 1.92 min; MS calculated value: 897.4; MS measured value: 898.5 [M+HT]. ¹H NMR (400 MHz, DMSO-ds) 8.29 (dJ = 8.2 Hz, 1H), 7.94 (t, J = 5.3 Hz, 1H), 7.94 (t, J = 5.3 Hz, 1H).1H), 7.91 — 7.86 (m, 1H), 7.84 (d,J= 8.5 Hz, 1H), 7.65 (t, J= 5.8 Hz, 25 —'1H), 7.35 (s, 1H), 7.14 (q, J= 8.1 Hz, 5H), 6.72 (s, 1H), 6.34 (s, 1H), 5.51 — 5.02 (m, 6H), 4.47 (td, J = 9.0, 4.6 Hz, 1H), 4.08 (s, 1H), 3.85 (s, 1H), 3.72 — 3.52 (m, 4H), 3.51 (d, J = 6.0 Hz, 2H), 2.95 (dd, J = 14.0, 4.6 Hz, 1H), 2.69 (dd, J = 13.5, 9.4 Hz, 1H), 2.54 (s, 3H), 1.88 (p, J = 6.9 Hz, 2H), 1.36 (s, 9H), 1.05 (d, J = 10.3 Hz, 9H), 0.89 (t, J = 7.4 Hz, 3H). Step 3: Compound 23c (16 mg, 0.018 mmol AF) was refluxed with 1 mL DCM, JHA 0.3 mL TFA, and At 30°C for 0.5 hours. LCMS HER, IER, WRIA. MAS ZK pL, 0.09 mmol D and compound 23d (11 mg, 0.036 mmol D) were purged with nitrogen three times. Add 1.0 mL of DMF to dissolve, stir for 1 second at room temperature (76 WO 2024 / 255740 PCT / CN2024 / 098491) for 1 hour. LCMS $d best. Pia AS 2 Pl ti WET), VRS ET PK, compound STI-A3, is a white solid 11 mg, yield 62.3%. LCMS (254 nm) purity 96.87%, Rt = 1.81 min; MS calculated value: 990.4; MS measured value: 991.1 [M+H]>. 4H NMR (400 MHz, DMSO-ds) 5 8.28 (d 1H), 7.87 — 7.81 (m, 3H), 7.35 (s, 1H), 7.22 (s, 2H), 7.21 — 7.13 (m, 2H), 7.21 — 7.13 (m, 2H), 7.28 (d 1H), 7.87 — 7.81 (m, 3H), 7.35 (s, 1H), 7.22 (s, 2H), 7.21 — 7.13 (m, 2H), 7.22 — ... 6H), 6.94 (s, 1H), 6.34 — 6.29(m, 1H), 5.42 5 —5.30(m, 6H), 4.47 - 4.45 (m, 1H), 3.69 — 3.64 (m, 4H), 3.57 (d, J= 6.1 Hz, 2H), 3.49 (d, J= 6.1 Hz, 2H), 3.37 (t, J= 7.1 Hz, 2H), 2.94 (s, 1H), 2.54 (s, 3H), 2.21 — 2.16 (m, 2H), 1.90 — 1.85 (m, 2H), 1.50 — 1.46 (m, 6H), 1.05 (s, 9H), 0.92 (t, J= 7.8 Hz, 3H). Intermediate compound 23b Preparation as follows: H @ 9 1) HATU, EtaN, DCM H 9 po an + TK 2) LIOH, MeOH soain Cy : OH 23b-1 23b-2 23b 10 Compounds 23b-1 (1.0 g, 4.31 mmol), 23b-2 (930 mg, 4.31 mmol), and HATU (2.458 g, 6.47 mmol) were added to a reaction flask. The mixture was purged with nitrogen three times. 20 mL of DCM was added to dissolve the compounds. Triethylamine (2.4 mL, 17.24 mmol) was added, and the mixture was stirred at room temperature for 7.0 hours. The reaction was monitored by LCMS until completion, and then stopped. The mixture was concentrated under reduced pressure, and EA and water were collected. PE and DCM were mixed. The mixture was filtered, and the solid was dissolved in methanol. The mixture was cooled in an ice bath, and LiOH (310 mg, 12.93 mmol, 0°C) was added. Stir for 30 minutes, then move to room temperature and stir for 3 hours. LCMS indicates the reaction is complete. Dry under reduced pressure, adjust pH to 2-3 with 1M hydrochloric acid, add EA and water, dry under anhydrous sodium sulfate, and dry under reduced pressure again to give compound 23b as a white solid, 694 mg, yield 53.6%. LCMS (214 nm, purity 97.25%, Rt = 1.25 min; MS calculated value: 379.2; MS measured value: 378.2) [MH]. ¹H NMR (400 MHz, DMSO-d.) 12.70 (s, 1H), 8.15 (d, J = 8.1 Hz, 1H), 7.93 (t, J = 5.7 Hz, 1H).7.28 (t, J=7.4 Hz, 2H), 7.21 (d J=7.5 Hz, 3H), 6.99 (t, J=6.0 Hz, 1H), 4.42 (td, J=8.5, 5.0 Hz, 1H), 3.69 (qd, J = 16.8, 5.6 Hz, 2H), 3.55 (d, J=6.0 Hz, 2H), 3.05 (dd, J = 13.8, 5.1 Hz, 1H), 2.87 (dd, J=13.8, 9.0 Hz, 20 Hz, 1H), 1.38 (s, 9H). Intermediate compound 23d was prepared as follows: [The remaining text appears to be a garbled string of characters and symbols and is not translated.] 23d-1 23d-2 23d UK PEAY 23d-1 (500 mg, 2.37 mmol) and compound 23d-2 (273 mg, 2.37 mmol) 8 F ZAR, added DCC (489 mg, 2.37 mmol), stirred at 0*C under nitrogen protection for 2 hours, then allowed to return to room temperature, and reacted overnight. The reaction was monitored by TLC (PE:EA 25”=11, product Rf=0.5) to indicate completion, and the forward reaction was stopped. The mixture was filtered, and the filtrate was subjected to silica gel column chromatography (PE:EA = 4:1~10D), concentrated to dryness under reduced pressure, to give compound 23d as a white solid product of 548 mg, yield 75.1%. ¹H NMR (400 MHz, DMSO-ds) 5 7.00 (s, 2H), 3.39 (t, J = 7.0 Hz, 2H), 2.81 (s, 4H), 2.65 (t, J = 7.3 Hz, 2H), 1.62 (p, J = 7.4 Hz, 2H), 1.51 (h, J = 7.2 Hz, 2H), 1.31 (q, J = 8.0 Hz, 2H). 30 “Example 24: Synthesis of drug linker conjugate STI-F3 77 WO 2024 / 255740 PCT / CN2024 / 098491 CR Lio SPQ Bost This” {tBuOK,Pd(OAc)p,Xantphos BocHN. Oo HaC and NAVY pb —— 4:6 a fF 'o TT = N . Na MeOH / DCM . N woreToulene, 100°C ef 9b 24a 2 reo ee ee 0 9 Step 1: 9% (1046 mg, 2.3 mmol / D) of compound 9 from Example 9 was added to a mixed bath of 60 mL dioxane and 60 mL methanol, followed by the addition of lithium oxide (166 mg, 6.9 mmol / D). The mixture was reacted at room temperature for 1 hour. The system was concentrated to dryness under reduced pressure, and the pH was adjusted to 3-4 with 1M hydrochloric acid. The mixture was AUGER 4 times, and the organic phase was concentrated to obtain compound 24a, a brownish-red solid. Approximately 1.0 g was measured. LCMS (254 nm) showed a purity of 93.42%, Rt = 1.88 min; MS calculated value: 414.1; MS measured value: 415.0. [M+H]*. 'H NMR (400 MHz, DMSO-de) 8.22 (d, J= 8.0 Hz, 1H), 7.98 (d J = 10.6 Hz, 1H), 7.31 (s, 1H), 6.56 (s, 1H), 5.44 (s, 2H), 5.25 (s, 2H), 2.54 (s, 3H), 1.87 (hept, J = 7.0 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H). Step 2: Dissolve compound 24a (200 mg, 0.48 mmol), All Boc-HA BEHK (1 26 mg, 0.72 mmol / L) in 6 mL of 10 FUK ASR, AERP FAT F108 mg, 0.96 mmol), Xantphos (28 mg, 0.048 mmol / L) and ethyl W411 mg, 0.048 mmOD), stirred thoroughly, heated to 100°C, and reacted for 3.5 hours. TLC (DCM:MeOH = 15:1, product Rf = 0.2) was used to detect the reaction. After completion, the system was concentrated to dryness under reduced pressure, purified by 1 M HRP A GAR, RS AHL, and silica gel column chromatography to obtain 39 mg of compound 24b as a red solid. 40 mg of starting material 24a was recovered, yielding a yield of 67.0%. ¹H NMR (400 MHz, DMSO-ds) 10.80 (s, 1H), 8.17 (d, J = 8.1 Hz, 1H), 7.85 (d, J = 8.1 Hz, 1H), 10.80 (s, 1H), 8.17 (d, J = 8.1 Hz, 1H), 7.85 (d, J = 8.1 Hz, 1H), 7.85 (d, J = 8.1 Hz, 1H).10.7 Hz, 1H), 157.30 (s, 2H), 6.52 (s, 1H), 5.42 (s, 2H), 5.03 (s, 2H), 4.04 (d, J = 6.0 Hz, 2H), 2.50 (s, 3H), 1.91 — 1.81 (m, 2H), 1.44 (s, 9H), 0.88 (t, 7 = 7.3 Hz, 3H). Step 3: Compound 24b (39 mg, 0.07 mmol HF) was added to 1 mL of dioxane under nitrogen protection with 0.3 mL of trichloroacetic acid. The mixture was stirred at room temperature for 30 minutes. After the reaction was completed by LCMS, the system was concentrated to dryness under reduced pressure. The crude product was then dilute with 1 mL of anhydrous DMF (Yt, AARP, DAS GAR, 0.71 mmol). 23b (27 mg, 0.07 mmol) was thawed with 20 HATU (41 mg, 0.11 mmol) for 1.5 hours. The product was then washed three times with LCMS Rel sen, We-- DMF, ZAR ZR and water tea, dried over anhydrous sodium sulfate, and evaporated to dryness to give 50 mg of compound 24c, a pale pink product, with a yield of 87.2%. LCMS (254 nm, purity 85.45%, Rt = 1.71 min; MS calculated value: 813.3; MS measured value: 814.1) [M+H]'. 'H NMR (400 MHz, DMSO-d6) 5 10.80 (s, 1H), 8.56 (t, J = 5.8 Hz, 1H), 8.53 (dd, J = 8.4, 1.4 Hz, 1H), 8.24 (s, 1H), 8.21 (d, / = 8.6 Hz, 1H), 7.91 (d, J = 10.7 Hz, 2H), 7.32 (s, 25 1H), 7.29 — 7.23 (m, 5H), 6.52 (s, 1H), 5.42 (s, 2H), 5.10 (s, 2H), 4.66 — 4.58 (m, 1H), 4.22 (d, J= 6.4 Hz, 2H), 3.78 (dd, J = 16.6, 5.7 Hz, 1H), 3.61 (dd, J = 17.4, 4.7 Hz, 1H), 3.53 (d,J= 5.7 Hz, 2H), 2.52 (s, 3H), 1.89 — 1.82 (m, 2H), 1.36 (s, 9H), 0.87 (s, 3H). 78 WO 2024 / 255740 PCT / CN2024 / 098491 Step 4: Compound 24c (45 mg, 0.055 mmol) was stirred at room temperature for 0.5 hours under LCMS protection (1.0 mL DCM #, 0.4 mL TFA, A). The reaction was monitored for completion by LCMS, the reaction was stopped, and the mixture was concentrated to dryness under reduced pressure. Triethylamine (76 pL, 0.550 mmoD) and compound 23d (34 mg, 0.110 mmoD) were added, and the mixture was purged with nitrogen three times. 1.0 mL of DMF was added to dissolve the compound, and the mixture was stirred at room temperature for 2.5 hours. The reaction was confirmed by LCMS. The reaction was then prepared under medium-high pressure reverse reaction (alkali mobile phase) to yield 11 mg of the drug linker 5 {KY STI-F3, a yellow solid. LCMS (254 nm, purity 100%, Rt = 1.64 min; MS+) (4: 906.3; MS measured: 907.0 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 8 10.79 (s, 1H), 8.53 (t, J = 5.7 Hz, 1H), 8.20 (dd, J = 8.5, 2.7 Hz, 1H). 2H), 8.06 (t, J= 5.8 Hz, 1H), 8.01 (t, J=5.4 Hz, 1H), 7.90 (d, J= 10.8 Hz, 1H), 7.32 (s, 1H), 7.31 — 7.21 (m, 5H), 6.97 (s, 2H), 6.52 (s, 1H), 5.42 (s, 2H), 5.09 (s, 2H), 4.60 (td, J= 8.9, 4.3 Hz, 1H), 4.24 (d, J= 7.9 Hz, 2H), 3.77 (dd, J= 16.7, 5.8 Hz, 2H), 3.71 — 3.55 (m, 6H), 10 2.51 (s, 3H), 2.09 (t, J= 7.5 Hz, 2H), 1.94 — 1.78 (m, 2H), 1.46 (t, J = 8.0 Hz, 6H), 0.88 (t, = 7.4 Hz, 3H).Example 23, Synthesis of Drug Linker Conjugate STI-F4 3 - 1 ow YK so BocHN IR oy 3 9 ° ° O 15 Step 1: The substrate was dissolved in compound 25a (4.3 g, 12.2 mmol OD), lead tetramethyl ester (6.8 g, 14.7 mmol OM), Al ARE OB LA VY Se (120.0 mL), and methyl methacrylate (40.0 mJ). Pyridine (1.16 mL, 14.7 mmol) was added, and the mixture was heated to 85°C and incubated for 3 hours. The reaction was then stopped. The solution was filtered through a silica gel filter, dried over EA AKER and ALA anhydrous Na2SO4, concentrated to dryness under reduced pressure, and purified by column chromatography (DCM:MeOH=50:D) to give 4.4 g of compound 2Sb as a white solid, yield 98.0%. LCMS (254 nm) purity 97.2%, Rt = 1.789 min; MS calculated value: 368.1; MS 20" measured value: 386.1 [M+NH4. Step 2: Compound STI-F2 (60 mg, 0.13 mmol), 25b (150 mg, 0.39 mmol / D) and p-methyl ATE ARILE WE FR (2 mg, 0.01 mmol / D) from Example 9 were added to a reaction flask, along with DCE (6.0 mL), AAR, and Fri. The reaction was carried out overnight at 80°C. The system was concentrated to dryness under reduced pressure and purified by Pre-TLC (DCM:MeOH = 15:1D) to obtain 40 mg of compound 2Sc as a yellow solid, with a yield of 34.1%. LCMS (254 nm) purity 93.2%, Rt = 1.811 min; MS calculated value: 761.2; MS 25 = SEMEL: 762.3 [M+H]. Step 3: Dissolve compound 25c (40 mg, 0.053 mmol D) in DMF (0.8 mD), add piperidine (0.2 mL), react at room temperature for 20 minutes, concentrate the system to dryness, dilute with water, freeze-dry, and add 23b (21 mg, 0.056 mmol), HATU (30 mg, 0.08 mmol D), and DMF (1.0 mL).Under vacuum protection, triethylamine (16 mg, 0.159 mmol) was added, and the reaction was stopped after TW for 1 hour. The system was concentrated to dryness, extracted with EA and saturated brine, and the aqueous phase was washed with EA 30 (10.0 mLx). The organic phases were combined, dried over anhydrous NasSO4, and concentrated to dryness under reduced pressure to give 30 mg of compound 25d, 79 WO 2024 / 255740 PCT / CN2024 / 098491 AYR IE Us, yield 62.9%. LCMS (254 nm, purity 97.2%, Rt = 1.662 min; MS calculated value: 900.3; MS measured value: 901.4 [M+H]". WHE A: Compound 25d (30 mg, 0.033 mmol JF) was added to 1.0 mL DCM, Under nitrogen protection, 0.3 mL of GA, Bin FD 20 EP was added. The reaction was monitored by LCMS until complete, then the reaction was stopped. The system was concentrated to dryness under reduced pressure. 1.0 mL of DMF was added to the system, and under nitrogen protection, triethylcarbamate (22 mg, 0.22 mmol / D) and 23d (10 mg, 0.033 mmol / D) were added sequentially. The reaction was carried out at room temperature for 20 min, then the reaction was stopped. The system was concentrated to dryness under reduced pressure and prepared by high-pressure reverse-phase reaction (acidic mobile phase). The mixture was frozen to obtain 6 mg of drug linker conjugate STI-F4, a yellow oily liquid with a yield of 27.9%. LCMS (254 nm) showed a purity of 98.65%, Rt = 1.591 min; MS calculated value: 993.4; MS measured value: 994.4. [M+H]*.'H NMR (400 MHz, DMSO-ds) 8 10. 74(s, 1H), 8.75 (t, J = 6.4 Hz, 1H), 8.37 (t, J = 6.0 Hz, 1H), 8.15 (d, J = 8.0 Hz, 10 1H), 8.07 (t, J= 6.0 Hz, 1H), 8.02 (t, J= 5.6 Hz, 1H), 7.90 (d, J = 10.8 Hz, 1H), 7.32 (s, 1H), 7.27 - 7.14(m, 6H), 6.98 (s, 2H), 6.52 (s, 1H), 5.42 (s, 2H), 5.15 (s, 2H), 4.84 — 4.77 (m, 2H), 4.52 — 4.47 (m, 1H), 4.38 (s, 2H), 3.80 (t, J = 6.0 Hz, 2H), 3.73 — 3.57 (m, 6H), 3.04 (dd, J = 14.0, 4.4 Hz, 1H), 2.82 — 2.79 (m, 1H), 2.50 (s, 3H), 2.09 (t, J = 7.6 Hz, 2H), 1.92 — 1.80 (m, 2H), 1.48 — 1.41 (m, 4H), 1.20 — 1.32 (m, 2H), 0.88 (t, J = 7.2 Hz, 3H). 15 Example 26: Synthesis of drug linker conjugate STI-G 9 HoN Fmoc-Gly-OH N l ne 1c fe) 20% Piperidine / DMF ord 1) cM) os 9 rt ha F " ss 20 ay TFA ae ie) F " SS Oo 9 ME 5 Q wo © F 、 Hof: “© Step 1: Compound 11f (200 mg, 0.89 mmol)#ll Fmoc-Gly-OH (530 mg, 1.78 mmol) ¥#-F 5 mL PU AMR, lal SC DARA UIA EtsN (360 mg, 3.56 mmol). HOBt (240 mg, 1.78 mmolD) and EDCI 20 (340 mg, 1.78 mmolD). The reaction mixture was stirred at room temperature. After the reaction was completed by TLC monitoring, 10 mL of ethyl acetate was added back to the reaction system for dilution. Water was added and the aqueous phase was collected. The aqueous phase was collected twice more with 10 mL of ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (PE:EA = 2:1). After evaporation to dryness, 300 mg of compound 26a was obtained as a pink solid, with a yield of 67%. LCMS (254 nm) Rt = 2.112 min; MS calculated value: SO3.2; MS measured value: 504.2.[M+H]*. 'H NMR (400 MHz, DMSO-ds) 9.45 (s, 1H), 8.07 (d, J = 8.8 Hz, 1H), 25 7.90 (d, J= 7.6 Hz, 2H), 7.73 (d, J= 7.5 Hz, 1H), 7.61 (s, 1H), 7.42 (t, J= 7.5 Hz, 2H), 7.34 (d, J= 80 WO 2024 / 255740 PCT / CN2024 / 098491 7.1 Hz, 3H), 6.57 (d, J = 12.9 Hz, 1H), 4.34 — 4.18 (m, 3H), 3.82 (d, J= 6.1 Hz, 2H), 2.81 (t, J=7.4 Hz, 2H), 1.58 (t, J=7.3 Hz, 2H), 1.35 — 1.19 (m 6H), 0.90 — 0.82 (m, 3H). Step 2: Compound 26a (200 mg, 0.40 mmol AF) and compound lc (105 mg, 0.40 mmol AF) were added to 4 mL of anhydrous methyl sulfoxide, and under nitrogen protection, refluxed for 30 minutes using a water separator. Then, p-methylthiazolic acid (18 mg, 0.1 mmol) was added, and ARSE 5 was added and stirred for 3.5 hours. The reaction was monitored by TLC (DCM:MeOH=10:1, product Rf=0.3) and LCMS. After the reaction was complete, the reaction system was cooled to room temperature, the reaction solution was evaporated to dryness, and then slurried with acetone and filtered to obtain 200 mg of the product. Compound 26b is a yellow solid in 70% yield. LCMS (254 nm) purity 96.96%, Rt = 2.084 min; MS calculated value: 730.3; MS measured value: 731.3 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 5 10.21 (s, 1H), 9.00 (d, J = 8.4 Hz, 1H), 8.01 (d, J = 12.0 Hz, 1H), 7.90 (d, J = 7.6 Hz, 2H), 7.75 (d, J = 7.8 Hz, 2H), 7.49 — 7.41 (m, 2H), 7.37 — 7.28 (m, 3H), 7.11 (d, 10 J = 7.9 Hz (1H), 5.43 (s, 2H),5.28 (s, 2H), 4.37 — 4.24 (m, 3H), 4.02 (d, J = 6.0 Hz, 2H), 3.11 (t, J = 8.0 Hz, 2H), 2.28 (s, 1H), 1.86 (p, J = 7.1 Hz, 2H), 1.72 (t, J = 7.7 Hz, 2H), 1.42 (d, J = 7.4 Hz, 2H), 1.34 (d, J = 7.1 Hz, 2H), 0.86 (d, J = 7.6 Hz, 6H). WHR 3: Compound 26b (50 mg, 0.068 mmol) was dissolved in 2 mL of 20% piperazine / DMF. The reaction mixture was stirred for 30 minutes, and TLC was performed. After the reaction was completed (DCM:MeOH=10:1, product Rf = 0.2), the reaction solution was directly purified by silica gel column chromatography (DCM:MeOH=20:1) to obtain 20 mg of compound 26c as a white solid, with a yield of 57.8%. LCMS (254nm) Rt = 1.670 min; MS calculated value: 508.2; MS measured value: 509.3 [M+H]*. Step 4: Compound 26c (20 mg, 0.039 mmol) was added to 1 mL of DCM / DMF (9:1D) mixture, and 23b (30 mg, 0.078 mmol), HATU (30 mg, 0.078 mmol), and DIPEA (25 mg, 0.195 mmol) were added. The reaction was carried out at room temperature with stirring for 30 minutes. After the reaction was completed, 1 mL of trifluoroacetic acid was added to the reaction mixture at room temperature and stirred for 4 hours. The crude product was prepared by reverse mixing under medium-high pressure (acid mobile phase) and lyophilized to obtain 10 mg of the compound as a yellow solid, with a yield of 33.3%. LCMS (254 nm) Rt = 1.616 min; MS calculated value: 769.3; MS measured value: 770.4 [M+H]*. 'H NMR (400 MHz, DMSO-d.)5 10.23 (s, 1H), 8.99 (d, J= 8.4 Hz, 1H), 8.58 (t, J= 6.0 Hz, 1H), 8.50 (t, J= 5.6 Hz, 1H), 8.36 (d, J = 8.4 Hz, 1H), 8.02 (d, J= 12.1 Hz, 1H), 7.98 (s, 2H), 7.28 (t, J= 7.7 Hz, 5H), 25 7.18 (t, J= 6.9 Hz, 1H), 6.52 (s, 1H), 5.43 (s, 2H), 5.29 (s, 2H), 4.65 (td, J= 9.2, 4.3 Hz, 1H), 4.12 (d, J = 5.7 Hz, 2H), 3.89 (dd, J = 16.8, 5.8 Hz, 1H), 3.71 (dd, J = 16.8, 5.3 Hz, 1H), 3.57 (s, 2H), 3.12 (q, J = 5.7, 4.9 Hz, 3H), 2.81 (dd, J = 13.8, 9.8 Hz, 1H), 1.86 (p, J = 7.2 Hz, 2H), 1.74 (t, J = 7.7 Hz, 2H), 1.41 (ddd, J = 21.6, 11.3, 5.2 Hz, 4H), 0.87 (td, J = 7.1, 3.8 Hz, 6H). WHR S: Compound 26d (20 mg, 0.026 mmol) F was added to 0.2 mL of DMF solution, along with 23d (16 mg, 30 0.052 mmol) Ill DIPEA (14 mg, 0.104 mmol D). The reaction was carried out at room temperature with stirring for 30 minutes. After the reaction was completed, the reaction solution was directly prepared by reverse high-pressure (acid mobile phase) and lyophilized to obtain 6 mg of drug linker conjugate STI-G, a white solid with a yield of 23.9%. LCMS (254 nm) purity 99.08%, Rt = 1.693 min; MS calculated value: 962.4; MS measured value: 963.1 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 8 10.19 (s, 1H), 9.00 (dd, 7= 8.5, 3.4 Hz, 1H), 8.45 (dt, J= 9.2, 5.8 Hz,3H), 8.14 (d, J= 8.3 Hz, 1H), 8.07 (t, J=5.7 Hz, 1H), 8.04 — 7.98 (m, 2H), 7.31 — 7.24 (m, 5H), 7.21 — 7.15 (m, 1H), 6.98 (s, 1H), 6.53 (d J = 7.8 Hz, 1H), 5.41 (d J= 14.1 Hz, 2H), 5.31 (d, J= 11.2 Hz, 2H), 4.62 — 4.50 (m, 2H), 4.11 (d, J =5.8 Hz, 2H), 3.74 (d J= 6.2 Hz, 1H), 3.67 (d, J = 5.7 Hz, 2H), 3.60 (d, J = 11.3 Hz, 1H), 3.11 (dd, J = 13.1, 5.2 Hz, 3H), 2.84 (dd, J = 13.8, 9.8 Hz, 1H), 2.10 (q, J= 6.1, 4.7 Hz, 2H), 2.06 — 1.93 (m, 2H), 1.86 (p, J =7.0 Hz, 2H), 1.77 — 1.70 (m, 2H), 1.50 (d, J= 7.0 Hz, 4H), 1.45 (dt, J= 7.7, 3.8 Hz, 4H), 0.89 — 0.84 (m, 6H). 81 WO 2024 / 255740 PCT / CN2024 / 098491 Example 27: Synthesis of Drug-linker Conjugate STI-G3 Can pect, Pd(AcO)₂, Cs₂CO₃ Xantphos DCM, DMF Toluene, 100°C F 1) 2) 236, HATU, TEA, DMF Subsequent 2) Ethylamine, 23d, DME ; BocHN─ ─ Step 1: Under nitrogen protection, compound STI-G2-06c of Example 13 (176 mg, 0.33 mmol), Boc-glycinamide (113 mg, 0.66 mmol) was added into 5 mL of anhydrous toluene, under nitrogen protection, cesium carbonate (215 mg, 0.66 mmol), Xantphos (19 mg, 0.03311Z.AR4E (7 mg, 0.033 mmol), PEPE, FRB 100C, reaction for 4.0 h. After the reaction was detected by LCMS, the system was concentrated to dryness under reduced pressure, dissolved in methanol, and prepared by reverse reaction under medium and low pressure (acid mobile phase) to give 45 mg of compound 27a as a standard solid, yield 21.7%. LCMS (254 nm) purity 96.8%, Rt = 1.86 min; calculated value: 626.2; measured value: 627.3 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 8 10.14 (s, 1H), 10 9.04 (d, J = 8.9 Hz, 1H), 8.07 (s, 1H), 8.04 (d, J = 11.9 Hz, 1H), 7.30 (s, 1H), 6.52 (s, 1H), 5.43 (t, 2H), 5.31 (s, 2H), 4.66 (s, 2H), 4.30 (s, 2H), 2.59 (t, J = 7.2 Hz, 2H), 1.87 (hept J = 7.1 Hz, 2H), 1.59 (q, J = 7.2 Hz, 2H), 1.42 (s, 9H), 0.93 — 0.89 (m, 3H), 0.89 — 0.86 (m, 3H). Step 2: Compound 27a (45 mg, 0.072 mmol) was added to 2 mL DCM, JHA, and 0.6 mL TFA, and stirred at room temperature for 1 hour under nitrogen protection. The reaction was monitored by LCMS until completion. The reaction was stopped, and the mixture was concentrated under reduced pressure. Triethylamine (73 mg, 0.72 mmol), compound 23b (27 mg, 0.072 mmol), and HATU (41 mg, 0.108 mmol) were added to the reaction flask. The mixture was purged with nitrogen three times, and 1.0 mL DMF was added for hydrolysis. The mixture was stirred at room temperature for 1.5 hours. LCMS The reaction was confirmed to be complete. The compound 27b was prepared under medium-high pressure reverse reaction (acidic mobile phase) and lyophilized to obtain 20 mg of compound 27b as a yellow solid, yield 31.4%. LC-MS (254 nm, purity 98.2%, Rt = 1.73 min; MS calculated value: 887.3; MS measured value: )888.3 [M+H]*.'H NMR (400 MHz, MeOD) 5 8.94 (d, J= 8.3 Hz, 1H), 7.65 (d, J= 11.8 Hz, 1H), 7.42 (s, 1H), 7.19 (dd, J= 4.5, 2.7 Hz, 6H), 7.10 (d, J= 6.3 Hz, 20 2H), 5.47 (d, J = 16.3 Hz, 1H), 5.26 (d, J= 16.2 Hz, 1H), 5.12 (d, J= 4.3 Hz, 1H), 4.85 (s, 1H), 4.65 — 4.60 (m, 2H), 4.49 (s, 2H), 4.13 (s, 2H), 4.05 (d, J = 11.3 Hz, 2H), 3.80 (s, 1H), 3.74 (s, 1H), 2.93 (dd, J = 14.0, 9.5 Hz, 2H), 2.52 (t, J = 7.3 Hz, 2H), 1.92 — 1.82 (m, 2H), 1.56 (p, J = 7.4 Hz, 2H), 1.33 (s, 11H), 0.94 — 0.88 (m, 3H), 0.88 — 0.83 (m, 3H). Step 3: Compound 27b (17 mg, 0.019 mmol) was placed in 1.0 mL DCM and protected with 0.4 mL TFA and AA 25. Stir at room temperature for 0.5 hours. Monitor the reaction progress using LCMS, then stop the reaction and concentrate to dryness under reduced pressure. Triethylamine (26 wL, 0.190 mmol) was added. F444 23d (12 mg, 0.038 mmoD) was added, purged three times with nitrogen, dissolved in 1.0 mL DMF, and stirred at room temperature for 1 hour. The reaction was confirmed by LCMS. A high-pressure reverse addition (acidic mobile phase) yielded 11 mg of the drug linker conjugate STIL-G3, a yellow solid. LCMS (254 nm, purity 100%, Rt = 1.65 min; MS calculated value: 980.4; MS measured value: 981.3 [M+H]). ¹H NMR (400 MHz, DMSO-ds) ¹⁵ 10.33 (s, 1H), 8.98 (d, J = 8.5 Hz, 1H), ³⁰ 8.10 (q, ⁻¹).J= 7.8 Hz, 2H), 8.05 (d, J = 11.6 Hz, 1H), 7.95 (s, 1H), 7.33 — 7.23 (m, 5H), 7.20 (s, 2H), 7.01 (s, 2H), 6.67 (s, 1H), 6.54 (d, J= 2.2 Hz, 1H), 6.40 (d, J= 2.4 Hz, 1H), 5.39 (s, 2H), 4.52 (d, J 82 WO 2024 / 255740 PCT / CN2024 / 098491 = 23.6 Hz, 2H), 4.32 — 4.25 (m, 1H), 3.89 (t, / = 12.3 Hz, 2H), 3.83 — 3.73 (m, 2H), 3.63 (t, J= 12.4 Hz, 2H), 3.14 (d, J= 11.8 Hz, 1H), 2.94 (d, J= 12.9 Hz, 1H), 2.89 (s, 2H), 2.73 (d, J= 2.3 Hz, 2H), 2.10 (d, J= 7.7 Hz, 2H), 2.00 (t, J= 7.4 Hz, 2H), 1.84 (q, J= 7.2 Hz, 2H), 1.57 (dd, J= 7.2, 3.7 Hz, 2H), 1.18 (t, J= 8.0 Hz, 4H), 0.87 (t, J= 7.3 Hz, 6H). 5 Example 28: The synthesis of drug linker conjugate STLF1002 is similar to that of STI-F4. The drug linker conjugate STI-F1002 was prepared as shown in the following procedure: Oe 1°. Step 1: Add compound STLF6 (40 mg, 0.09 mmol) and 25b (40 mg, 0.10 mmol) to a reaction flask, add 12-dichloroethane (C2.0 mL), and incubate overnight at 80°C. Concentrate the system under reduced pressure to dryness and purify by Prep-TLC (DCM:MeOH=15:1) to obtain 46 mg of compound 28a, a yellow solid with a yield of 69.7%. LCMS (254 nm) purity 87.90%, Rt =1.90 min; Calculated value: 775.3; Measured value: 776.2 [M+H]'« Step 2: Add piperidine (0.4 mL) to compound 28a (46 mg, 0.06 mmol) and DMF (1.8 mD), concentrate the system to dryness for 20 minutes, add (butyroxyl)glycyl-L-alanine (23b) (27 mg, 0.07 mmolD), 2-(7-azadecano-1-yl)-NJN'N-tetramethylurea hexafluorophosphate (34 mg, 0.09 mmolD) and NIN-dimethylformyl phosphate (1.0 mL) to the residue, under nitrogen protection, add triethylamine (16 mg, 0.159 mmolD), and react at room temperature for 1 hour. PELE BOD, the system was concentrated to dryness, dichloromethane and saturated brine were added and collected, the aqueous phase was washed with dichloromethane (10.0 mLx), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure to give 86 mg of compound 28b Hin, 20" as a yellow oily liquid, yield 62.9%. LCMS (254 nm, purity 93.41%, Rt = 1.4 min; calculated value: 914.4; measured value: 915.4 [M+H]-). Step 3: Compound 28b (86 mg, 0.10 mmol) was dissolved in dichloromethane (1.8 mL), RAR, SFR (0.1 mL), JHAE HH, SH PRY for 1 hour, and the reaction was stopped when the LCMS was monitored until complete. The system was concentrated to dryness under reduced pressure, and NJN-dimethylformyl methyl ether (2.0 mD) was added. Under nitrogen protection, triethyl gum (111 mg, 1.10 mmol / L), 6-(maleic succinimide) hexanoate succinimide (EMCS, 52 mg, 0.17 mmol / L), and 4 μmol / L were added sequentially. The reaction was stopped after 4 hours, and the system was concentrated to dryness under reduced pressure. The mixture was then directly prepared by high-pressure reverse-phase reaction (mobile phase: ethyl acetate / 0.05% trifluoroacetic acid).The aqueous solution was lyophilized to give 12 mg of compound STI-F1002 as a yellow solid, with a yield of 12.6%. LCMS (254 nm, purity 98.5%, Rt = 1.58 min; MS calculated value: 1007.4; MS measured value: 1008.4 [M+H]T). ¹H NMR (400 MHz, DMSO-ds) 510.05 (s, 1H), 8.50 (t, J = 6.6 Hz, 1H), 8.41 (d, J = 5.9 Hz, 1H), 8.29 (t, J = 5.9 Hz, 1H), 8.11 (d, J = 8.0 Hz, 1H), 8.07 (t, J = 5.8 Hz, 1H), 8.01 (t, J = 5.8 Hz, 1H), 7.59 (d, J = 10.5 Hz, 1H). 7.27 — 7.16 (m, 5H), 7.00 (s, 1H), 6.99 (s, 2H), 6.61 (br, 1H), 5.49 (s, 2H), 5.43 (s, 2H), 4.54 — 4.44 (m, 83 WO 2024 / 255740 PCT / CN2024 / 098491 1H), 3.76 — 3.56 (m, 10H), 3.39 — 3.33 (m, 4H), 3.04 (d, J= 4.8 Hz, 1H), 3.01 (d, J = 4.7 Hz, 1H), 2.44 (s, 3H), 2.13 (t, J= 7.4 Hz, 2H), 1.91 — 1.80 (m, 2H), 1.69 — 1.60 (m, 2H), 1.50 — 1.41 (m, 4H), 1.26 — 1.14 (m, 4H), 0.87 (t, J = 7.3 Hz, 3H). 5” Example 29: Synthesis of drug linker conjugate STLF1001 pg TO ay g hye Step 1:; Compound STILF (80 mg, 0.16 mmol), 25b (61 mg, 0.16 mmol), Fl Xf FAA HER Me EE (4 mg, 0.01 mmol JIA RNG, DIA 1,2-— AZ. 665.0 mL), BAR, FZ 7$*C reacted for 48 hours, then the system was concentrated to dryness under reduced pressure and measured by Prep-TLCPurification (DCM:MeOH=15:1) yielded 40 mg of compound 10 FI001-01, Wie IAPR, PEFR 31.3%. LCMS (254 nm, purity 92.16%, Rt = 1.905 min; calculated value: 801.3; measured value: 802.1 [M+H]*. Step 2: Compound F1001-01 (40 mg, 0.05 mmol) was mixed with NJN-dimethylformamide (0.8 mL), HLA piperidine (0.2 mL), and reacted at room temperature for 20 minutes. The system was then concentrated to dryness. (S-butyroxyl)glycyl-L-decylalanine (23b) (27 mg, 0.07 mmol), 2-(7-azabenzotriazol-1-yl)-NNN"N-tetramethyladenosylhexane phosphate 15 (34 mg, 0.09 mmol), NIN-dimethylformamide (1.0 mL), AW. BIAS ZAKS mg, 0.15 mmol), +i FRM 1 JN. GARAIRSRE FE, WADA ER HAE BAAR, 7K AB LAA GE10.0 mL), GIFANLA, FOKGEMAFIR, TRAE, 2S] 46 mg Compound F1001-02 #14» LCMS (254 nm purity 72.13%, Rt = 1.708 min; calculated value: 940.4 Measured value: 941.2 [M+H]*. 20 Step 3: Add compound F1001-02 (46 mg, 0.05 mmol) to dichloromethane (1.8 mL), BARE. trifluoroacetic acid (0.1 mD), react at room temperature for 1 hour, monitor the reaction until complete by LCMS, and stop the reaction. Concentrate the system to dryness under reduced pressure, and add NJN-dimethylformamide (2.0 mD). Under nitrogen protection, triethylamine (0.07 mL, 0.50 mmol) and 6-(maleimide) hexanoic acid succinimide (EMCS, 5 mg, 0.17 mmol D) were added sequentially. The reaction was carried out at room temperature for about 1 hour. The reaction was then stopped, and the system was concentrated to dryness under reduced pressure.After preparation by high-pressure reversed-phase (mobile phase: acetonitrile / 0.05% trifluoroacetic acid aqueous solution 25 WM), 12 mg of compound STI-F1001 was obtained as a yellow solid with a yield of 23.7%. LCMS (254 nm): purity 96.79%, Rt = 1.58 min; MS calculated value: 1033.4; MS found value: 1034.1 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 10.69 (s, 1H), 8.72 (t, J= 6.6 Hz, 1H), 8.35 (t, J= 5.9 Hz, 1H), 8.12 (dd, J= 8.3, 4.3 Hz, 2H), 8.06 (t, J = 5.9 Hz, 1H), 8.00 (t, J = 5.8 Hz, 1H), 7.91 (d, J = 10.7 Hz, 1H), 7.33 (s, 1H), 7.26 — 7.11 (m, 5H), 6.98 (s, 2H), 6.53 (br, 1H), 5.41 (s, 2H), 5.14 (s, 2H), 4.86 — 4.71 (m, 2H), 4.48 (td, J = 8.9, 4.6 Hz, 1H), 3.83 (d, J = 7.5 Hz, 1H), 3.77 (d, J = 6.4 Hz, 2H), 3.75 — 3.53 (m, 6H), 3.02 (dd, J = 14.0, 4.4 Hz, 1H), 2.77 (dd, J = 13.9, 9.7 Hz, 1H), 2.50 (s, 3H), 2.09 (t, J = 7.6 Hz, 2H), 1.92 — 1.81 (m, 2H), 1.50 — 1.41 (m, 4H), 1.31 (q, J=6.4 Hz, 1H), 1.18 (dd, J= 15.4, 7.8 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H), 0.65 — 0.56 (m, 4H). 84 WO 2024 / 255740 PCT / CN2024 / 098491 Similarly, the drug-linker conjugates described below are prepared; Example No. 30 O, -NH> O O Oo H § N N_ — 9 Ni. N ‘ou: N — HOCy HG A a \ Oo O N Oo H3C STI-F1003 F 31 Ox -NH2 ye O O CY “CH 1 ow ( Lond \ O me | fe) fe) N STI-F1004 HC Example 32: Synthesis of Drug-linker Conjugate STI-F1006; ° No OS 5 ,} bod ° i o ped HN ot hoy oe oneal ——————— SC fo 六 二。 I F F1002-02 fad oO H Ox me Cyn a A eye a 5 STI-F1008 : Step 1: F1006-S1 (500 mg, 1.13 mmol) was dissolved in N,N-dimethylformamide (10.0 mL), under argon protection, then EMCS (382 mg, 1.24 mmol) and N,N-diisopropylethylamine (219 mg, 1.70 mmol) were added. The reaction was carried out at room temperature for 2 hours, quenched by adding water, and directly subjected to reversed-phase preparation (mobile phase: acetonitrile / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 587 mg of compound F1006-S2 was obtained as a pale yellow oily liquid with a yield of 80.7%. LCMS (254 nm) purity 98.8%, Rt=1.38 min; MS calculated value: 634.3; MS observed value: 635.2 [M+H]+. Step 2: F1006-S2 (578 mg, 0.93 mmol) and N,N'-dicyclohexylcarbodiimide (210 mg, 1.02 mmol) were dissolved in THF (12.0 mL), under nitrogen protection, then pentafluorophenol (188 mg, 1.02 mmol) was added, and the reaction was carried out at room temperature overnight. After LCMS monitoring indicated the completion of the reaction, the system was filtered by suction, and the filtrate was concentrated to dryness under reduced pressure to obtain 587 mg of compound F1006-S3 as a pale yellow oily liquid with a yield of 80.6%. LCMS (254 nm) purity 80.0%, Rt = 1.85 min; MS calculated value: 800.3; MS observed value: 801.3 [M+H]+. Step 3: Compound 28b (100 mg,Dissolve 0.11 mD in dioxane (1.8 mD) under nitrogen protection, add 0.2 mL of trifluoroacetic acid (AHHH) and frost at 0-5 °C for 3 hours. Monitor the reaction with LCMS until it is complete, then stop the reaction. The system was concentrated to dryness under reduced pressure (85 WO 2024 / 255740 PCT / CN2024 / 098491). NN-dimethylformamide (2.0 mL) was added, and the mixture was cooled to 0–5 °C under nitrogen protection. N,N-diisopropylethylamine (111 mg, 1.10 mmol) and F1006-S3 (80 mg, 0.10 mmol) were added sequentially. The reaction was allowed to proceed for approximately 0.5 hours, at which point the reaction was stopped. The system was then concentrated to dryness under reduced pressure and directly prepared by high-pressure reverse-phase reaction (mobile phase: acetonitrile / 0.05% trifluoroacetic acid aqueous solution). After freezing, 16 mg of compound STI-F1006 was obtained as a yellow solid, with a yield of 10.2%. LCMS (254 nm, purity 99.8%, Rt = 1.59 5 min; MS calculated value: 1430.7; MS measured value: 1429.5 [MH]-). IH NMR (400 MHz, DMSO-ds) 8 8.52 (t, J = 6.7 Hz, 1H), 8.30 (d, J = 7.5 Hz, 2H), 8.17 (d, J = 5.7 Hz, 1H), 8.07 (t, J = 5.7 Hz, 1H), 8.11 (d, J = 8.0 Hz, 1H), 8.01 (t, J = 5.8 Hz, 1H), 7.82 (t, J = 5.7 Hz, 1H), 7.58 (d, J = 11.0 Hz, 1H), 7.38 (t, J= 5.6 Hz, 1H), 7.23 — 7.18 (m, 5H), 6.99 (s, 2H), 6.48 (s, 1H), 5.44 (s, 2H), 5.41 (s, 2H), 4.51 — 4.45 (m, 1H), 3.77 — 3.68 (m, 6H), 3.66 — 3.57 (m, 8H), 3.50 (s, 30H), 3.19 — 3.16 (m, 4H), 3.05 (d, J= 10 4.8 Hz, 1H), 3.01 (d, J= 3.6Hz,1H), 2.43 (s, 3H), 2.38 (t, J = 6.5 Hz, 2H), 2.03 (t, J = 7.4 Hz, 2H), 1.88 — 1.81 (m, 2H), 1.66 — 1.64 (m, 2H), 1.50 — 1.43 (m, 4H), 1.24 — 1.18 (m, 4H), 0.87 (t, J = 7.3 Hz, 3H). Example 33: Synthesis of drug linker conjugate STI-F1010 Step 1: Compound F1007-01 (1.64 g, 4.00 mmol / L) was added to 50 mL of MAA GE, IA N,N-3 AlZé ZR (1551 mg, 12.00 mmol / L) under ice bath conditions. 525 mg (4.00 mmol D) of glycine tert-butyl ester (HT-Gly-OtBu, 525 mg, 4.00 mmol D) and 2-(1H-decanotriazo-L-1-yl)-,1,3,3-tetramethylurea tetrahydroboronic acid ester (1349 mg, 4.20 mmol D) were added to 20 mL of dioxane and 4 mL of trifluoroacetic acid. The mixture was reacted at room temperature for 1 hour under nitrogen protection. After the reaction was complete as monitored by LCMS, the mixture was filtered and washed several times with dichloromethane. The filter cake was added to 8 mL of 20" dioxane and 4 mL of trifluoroacetic acid was added. The mixture was reacted at room temperature for 3 hours. After the reaction was complete as monitored by LCMS, the mixture was evaporated to dryness. Compound F1010-01 was a white solid with a yield of 68.5%. LCMS (254 nm) showed a purity of 100%, Rt = 1.695 min. 467.2; MS measured value: 468.3 [M+H]*. Step 2: Dissolve the substrate of compound F1010-01 (545 mg, 1.17 mmol), HIA VU AVI (1 5.0 mD), and toluene (5.0 mL) under nitrogen protection. Add lead tetraacetate (620 mg, 1.40 mmol) and pyridine (1 L 11 mg, 1.40 mmol), FHA 25 85°C, RM 3 DIN, PIER. ERLE, VER RRA, RE iT ah (DCM:MeOH=100:1), #42!) 400 mg compoundF1010-02 is a white solid with a yield of 71.0%. LCMS (254 nm) purity is 89.0%, Rt = 1.811 min; MS calculated value: 481.2; MS measured value: 499.7 [M+NH4]. Step 3: Compound STI-F6 (160 mg, 0.34 mmol), F1010-02 (197 mg, 0.41 mmol), and pyridine p-methylthiosulfonate (PPTS, 10 mg) from Example 17 were added to a reaction flask. DMF (5.0 mL) was added, and the reaction was stopped after about 3 hours in an AAR. FE 30 "9SsC flask. The system was concentrated to dryness under reduced pressure and purified by column chromatography (DCM:MeOH=50:1D) to give 110 mg of compound F1010-03, AREZT GIA, yield 25.5%. LCMS (254 nm, purity 84.0%, Rt = 1.920 min; MS calculated value: 888.4; MS measured value: 889.2 [M+H]*. Step 4: Compound F1010-03 (55 mg, 0.062) was added to a reaction flask. mmol was added to DMF (0.5 mID), piperidine (0.05 mL) was added, and the mixture was reacted at room temperature for 20 4 h. RAAB, RR TS. UR FG, TAR PULA, EMCS (29 mg, 0.093 mmol D) and DMF (1.0 mL D) were added under choke protection. Triethylamine (19 mg, 0.19 mmol) was then added, and the mixture was reacted for 1 h. The mixture was concentrated to dryness and prepared under medium-high pressure reverse phase (mobile phase: acetic acid / 0.05% formic acid aqueous solution). After lyophilization, 16 mg of compound STI-F1010 was obtained as a pale yellow solid with a yield of 30.2%. LCMS (254 nm, purity 96.38%) was performed. 1.663 min; MS calculated value: 89.4; MS measured value: 860.4 [M+H]*. 'H NMR (400 MHz, DMSO-ds) 5 8.79 (t, J = 6.4 Hz, 1H), 8.59 (t, J = 6.6Hz, 1H), 8.42 (d, J=7.8 Hz, 1H), 8.00 (d, J= 7.2 Hz, 1H), 7.76 (t, J = 8.7 Hz, 1H), 7.59 (d, J= 10.5 Hz, 1H), 7.41 (s, 1H), 7.00 (s, 2H), 6.57 (br, 1H), 5.47 (s, 2H), 5.44 (s, 2H), 4.58 — 4.56 (m, 2H), 4.23 (q, J= 7.1 Hz, 1H), 4.14 — 4.09 (m, 1H), 3.71 (d, J= 6.8 Hz, 2H), 3.45 - 3.43 (m, 2H), 3.35 (t, J = 7.0 Hz, 2H), 2.44 (s, 3H), 2.16 — 2.05 (m, 2H), 1.88 — 10 1.84 (m, 1H), 1.77 — 1.72 (m, 2H), 1.67 — 1.61 (m, 2H), 1.47 — 1.42 (m, 4H), 1.24 — 1.12 (m, 7H), 0.88 (t, J = 6.0 Hz, 3H), 0.79 (dd, J = 11.3, 6.7 Hz, 6H). Example 34: Synthesis of drug linker conjugate STILF1011 15 Compound F1010-03 (55 mg, 0.062) from Example 33 was used. 0.5 mL of DMF (0.5 mmol), J AURHE (0.05 mLD) were added, and the mixture was reacted at room temperature for 10 minutes. The system was concentrated to dryness, and the residue was washed with petroleum ether, dried under reduced pressure, and then HATU (57 mg, 0.19 mmol), F1006-S2 (29 mg, 0.093 mmol) from Example 32, and DMF (1.0 mL) were added. The mixture was protected with nitrogen, and then triethylamine (19 mg, 0.19 mmol) was added. The mixture was reacted at room temperature for 0.5 hours, and the reaction was stopped. The system was concentrated to dryness and prepared under medium-high pressure reverse phase (mobile phase: ethyl acetate / 0.05% formic acid aqueous solution). After drying, 28 mg of compound 20 STI-FIO11 was obtained, which was a pale yellow viscous solid with a yield of 35.0%. LCMS (254 nm purity) was used to prepare the compound.96.07%, Rt = 1.597 min; MS calculated value: 1282.6; MS measured value: 1283.6 [M+HT .IH NMR (400 MHz, DMSO-d¢) & 8.60 — 8.56 (m, 2H), 8.49 — 8.45 (m, 1H), 8.03 (d, J= 7.3 Hz, 1H), 7.90 — 7.78 (m, 2H), 7.60 (d, J= 10.3, 1H), 7.49 (s, 1H), 7.00 (s, 2H), 6.53 (br, 1H), 5.51 (s, 2H), 5.45 (s, 2H), 4.62 — 4.52 (m, 2H), 4.23 (t, J = 7.2 Hz, 1H), 4.15 (dd, J= 8.7, 6.6 Hz, 1H), 3.75 — 3.73 (m, 2H), 3.60 — 3.52 (m, 4H), 3.51 — 3.46 (m, 30H), 25 3.39 — 3.36 (m, 4H), 3.16 (q, J = 5.9 Hz, 2H), 2.45 (s, 3H), 2.03 (t, J= 7.4 Hz, 2H), 1.88 — 1.84 (m, 1H), 1.78 — 1.73 (m, 2H), 1.68 — 1.62 (m, 2H), 1.50 — 1.42 (m, 4H), 1.23 — 1.12 (m, 7H), 0.87 (t, J = 7.4 Hz, 3H), 0.79 (dd, J = 11.5, 6.8 Hz, 6H). SH 35: Synthesis of drug linker conjugate STILF1012 alanana. TOL, 1) Pipercine, OME ° escianene ae 30 F1010-08 an: 1006.01 STLF 1012 med Compound F1010-03 (70 mg, 0.078 mmol) from Example 33, JAF DME (0.5 mL), JILAURHE (0.05 mL), were reacted at room temperature for 10 minutes. The system was concentrated to dryness, the residue was washed with petroleum ether, and dried under reduced pressure. HATU (87 mg, 0.23 mmol D), 6-(2-(methylsulfonyl) methyl-$-yl) hex-5-Triethylamine (25 mg, 0.094 mmol) and DME (2.0 mL) were added under nitrogen protection, followed by the addition of triethylamine (23 mg, 0.23 mmol). The reaction was stopped after 0.5 hours of FAIR treatment. The system was concentrated to dryness and prepared under medium-high pressure reverse phase (mobile phase: ethyl acetate / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 87 WO 2024 / 255740 PCT / CN2024 / 098491 30 mg 444% STI-F1012 was obtained as a pale yellow solid with a yield of 41.9%. LCMS (254 nm, purity 98.69%, Rt = 1.646 min; MS calculated value: 916.4; MS measured value: 917.3 [M+H]*.'H NMR (400 MHz, purity 98.69%, Rt = 1.646 min; MS calculated value: 916.4; MS measured value: 917.3) DMSO-ds) 6 9.11 (s, 2H), 8.60 (t, J = 6.6 Hz, 1H), 8.45 (d, J= 7.7 Hz, 1H), 8.06 (d, J= 7.2 Hz, 1H), 7.89 (d, J= 8.7 Hz, 1H), 7.58 (dd, J = 10.3, 2.3 Hz, 1H), 7.41 (s, 1H), 6.65 (br, 1H), 5.49 (s, 2H), 5.45 (s, 2H), 4.64 — 4.51 5 (m, 2H), 4.24 (q, J= 7.1 Hz, 1H), 4.16 (dd, J = 8.7, 6.7 Hz, 1H), 3.76 — 3.69 (m, 4H), 3.45 — 3.42 (m, 1H), 3.40 (s, 3H), 2.54 (d, J = 7.3 Hz, 2H), 2.44 (s, 3H), 2.16 — 2.05 (m, 2H), 1.89 — 1.83 (m, 2H), 1.81 — 1.76 (m, 4H), 1.67 — 1.61 (m, 2H), 1.21 (d, J = 7.1 Hz, 3H), 0.87 (t, J = 7.4 Hz, 3H), 0.81 (dd, J = 11.8, 6.7 Hz, 6H). 10 “Example 36: Synthesis of drug linker conjugate STILF1013 mee ee AT gee BF as aig Ee HE 1: F1013-01(2.0 g, 4.27 mmol) bleached in dioxane (20.0 mL), BAUR FRU A BUA phenol (943 mg, 5.12 mmol), DCC (1.05 g, 5.12 mmol), reacted at room temperature for 1 ME. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with dioxane (5 mL * 3). The filtrate was concentrated to dryness at room temperature and used directly for the next step. The above product was dissolved in acetone / water (5 mL / 15 mL), and sodium carbonate (1.07 g, 12.8 mmol / L) and glycine (320 mg, 4.27 mmol) were added sequentially. After tweezing for 2 hours, the pH was adjusted to 3-4 by adding 1M hydrochloric acid, followed by washing with ethyl acetate (10 mL*2), saturated brine (10 mL*2), drying with anhydrous sodium sulfate, and concentrated at 45°C. The product was then directly prepared as a white solid by medium-low pressure reverse phase reaction (Gist: Z.fs / 0.05% = CRIA), URT late). 2.0 g of compound F1013-03 was prepared, yielding 90.9%. LCMS (254 nm) FE 95.6%, Rt = 2.16 min; MS calculated value: S25.2, MS measured value: 5326.3 20 [M+H]*. Step 2: F1013-03 (2.0 g, 3.80 mmol) was mixed with Pyridine (40 mD) and methyl methacrylate (10 mL) under atmospheric protection. Then, Pb(OAc)s (2.03 g, 4.57 mmol) and pyridine (361 mg, 4.57 mmol) were added. The mixture was reacted at 8°C for 1 hour. After the reaction was complete, the reaction solution was concentrated to dryness under reduced pressure at 4°C. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 3) to give 1.80 g of compound F1013-04, a pale yellow solid, with a yield of 87.8%. LCMS (254 nm, purity 92.1%, Rt = 2.36, 25 min; MS calculated value: S 39.2; MS measured value: 357.3 [MHNH4]. Step 3: Add compound F1013-04 (260 mg, 0.43 mg) to the solution.mmol) was added to DMF (5 mD) under chlorine protection, and STI-F6 (170 mg, 0.36 mmol) and PPTS (17 mg) from Example 17 were added sequentially. The mixture was incubated at 95°C for 2 hours. The reaction solution was reduced to dryness under reduced pressure at 45°C. The crude product was purified by column chromatography (dichloromethane / methanol = 0.03) to give 70 mg of compound F1013-05, which was black (EK, PEFR 14.2%). LCMS (254 nm) showed a purity of 70.1%, Rt = 2.05 min; MS calculated value: 946.4; MS measured value 30 = {8}: 947.5 [M+H]-. Step 4: Compound F1013-05 (70 mg, 0.07 mmol) was added to DMF (0.7 mD). Add piperidine (70 WL), stir under nitrogen protection for 0.5 hours, and monitor the reaction until complete by LCMS. After concentrating the reaction solution to dryness using a 45°C oil pump, add petroleum ether 88 WO 2024 / 255740 PCT / CN2024 / 098491 and slurry (2 mL * 3). Concentrate again and use directly for the next step. Add F1013-06 (64 mg, 0.07 mmol) to DMF (2 mL), (KYRA HATU (42 mg, 0.11 mmol)#ll NN-diisopropylethylamine (19 mg, 0.148 mmol), and stir under nitrogen protection for 5 minutes. Then add the above concentrate, stir at room temperature under nitrogen protection for 1 hour, and monitor the reaction until complete by LCMS. After concentrating to dryness using a 45°C oil pump, the crude product is purified by column chromatography (dioxane / methanol = 0 / 7) to obtain 60 mg Compound F1013-07, 5”, is a yellow solid with a yield of 52.1%. LCMS (254 nm) showed a purity of 93.0%, Rt = 1.78 min; MS calculated value: 1565.1; MS measured value: 1366.2 [M+H]*. Step 5: Compound F1013-07 (60 mg, 0.03 mmol) was dissolved in 10% TFA / DCM (2 mLD) under nitrogen protection and stirred for 1 hour. LCMS monitoring was performed.The reaction was complete. The reaction solution was adjusted to pH 7-8 with triethylamine, purged with nitrogen, and then prepared under high pressure in reverse phase (mobile phase: triethylamine / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 6.2 mg of compound STI-F1013 was obtained as a yellow solid with a yield of 11.8%. LCMS (254 nm, purity 99.0%, Rt = 1.51 min; MS calculated value: 1465.6; MS measured value: 1466.8) [M+H]+. 1H NMR (400 MHz, MeOD) 8.95 (s, 2H), 8.32 (dJ = 7.6 Hz, 1H), 7.96 (s, 1H), 7.70 (s, 1H), 7.56 (d, J = 10.0 Hz, 1H), 5.66–5.58 (m, 2H), 4.77 (q, J = 10.4 Hz, J = 26.4 Hz, 2H), 4.56 (t, J = 4.8 Hz, 2H), 4.45–4.41 (m, 3H), 4.03 (s, 2H), 3.98 (d, J = 2.0 Hz, 2H), 3.89 - 3.83 (m, 4H), 3.64 - 3.57(m, 28H), 3.52 - 3.49 (m, 2H), 3.37 (s, 3H), 3.37 - 3.31 (m, 6H), 15 2.97 (t, J= 7.2 Hz, 2H), 2.60 (t, J= 7.2 Hz, 2H), 2.52 (s, 3H), 2.45 (t, J= 7.2 Hz, 2H), 2.03 - 1.68 (m, 12H), 1.51-1.47 (m, 2H), 1.02 (t, J= 7.2 Hz, 3H). Intermediate compound F 1013-06 was prepared as follows: Step 1: Compound 6-(2-(methanesulfonyl)carbin-(2-yl)hexyl(2-yl)carboxylic acid (200 mg, 0.74 mmol) + — AHH 20” and valerium (2 mD) were added sequentially to HATU (425 mg, 1.12 mmol) and NJN-diisopropylethylamine (193 mg, 1.49 mmol), and the mixture was stirred under nitrogen protection for 10 minutes. Then, 49 mg of propyl ester (49 mg, ...) was added to the mixture.Add 0.89 mmoD to the reaction solution and continue stirring for 1 hour. Monitor the reaction for completeness using LCMS. The reaction solution was concentrated to dryness under reduced pressure at room temperature. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 3) to give 210 mg of compound F1005-02 as a yellow solid, with a yield of 92.5%. LCMS (254 nm) showed a purity of 92.3%, Rt = 2.20 min; MS calculated value: 30.0; MS measured value: 306.1 [M+H]*. 25 Step 2: Compound F1005-02 (100 mg, 0.32 mmol) was dissolved in a mixed solvent of dimethyl phthalate (1 mL) and water (0.1 mD). F1005-03 (144 mg, 0.32 mmol) and CuBr (94 mg, 0.65 mmol) were added sequentially. The reaction was stirred under nitrogen protection for 2 hours, and the reaction was monitored by LCMS to ensure complete reaction. The reaction solution was prepared by low-pressure reverse phase reaction (mobile phase, Ethylene / 0.05% trifluoroacetic acid aqueous solution), after lyophilization, yielded 350 mg of compound F1013-06, a green oily substance. LCMS (254 nm) purity 85.2%, Rt = 1.95 min; MS calculated value: 89.3; MS measured value: 860.4 [M+H]°. 30 Example 37: Synthesis of drug linker conjugate STILF1014 89 WO 2024 / 255740 PCT / CN2024 / 098491 F 4 Ni eon 1) Fmoc-GGF-OH, Piperidine 3 Al -we we cs ° F1006-03 F1006-S3 9 9 9 Ce i ; and H “™ 1) Piperidine, DMF Cy ol O ~ al 2) F1014-01,HATU,DMF,DIPEA ° N H3C STI-F1014 F AEE 1 AY(QH-2-9-28) FASE) PRE) HA EL- AK WY SA BR (Fmoc-GGF-OH, 400 mg, 0.80 mmol) was dissolved in DMF (5.4 mD), piperidine (0.6 mLD) was added, and the mixture was reacted at room temperature for 30 minutes. The mixture was then concentrated to dryness.The residue was slurried with petroleum ether, and redissolved in DMF (8.0 mL), AR, PRIZE 0~5°C, DLA F1006-03 (832 5 mg, 1.04 mmol) DIPEA (154 mg, 1.20 mmol), and incubated at 0~5°C / RIVA for 1 hour. The reaction was monitored by LCMS until complete. (FIER. PRAMIRIRZA . MER AA, 2) 380 mg of compound F1006-S3 was obtained as a colorless oily liquid, with a yield of 40.8%. LCMS (254 nm, purity 95.96%, Rt = 1.648 min; MS calculated value: 895.4; MS measured value: 896.5 [M+HJ-. Step 2: Compound F1014-01 (180 mg, 0.22 mmol / D) was dissolved in DMF (2.7 mD), piperon (0.3 mL) was added, and the mixture was reacted at room temperature for 30 minutes. The system was concentrated to dryness, slurried with acetone, filtered, and the filter cake was redissolved in DMF (3.0 mL). F1006-S3 (63 mg, 0.07 mmol), HATU (125 mg, 0.33 mmol / D), and nitrogen protection were added. DIPEA (43 mg, 0.33 mmol) was added, and the mixture was reacted at room temperature for 0.5 hours. The reaction was monitored by LCMS until complete. The reaction was then stopped. The system was concentrated to FF, PRR RG AGRA: ZNR / 0.05%= HR CB KAR), HPAES 14 mg. Compound STI-F1014 was a pale yellow solid with a yield of 4.4%. LCMS (254 nm, purity 95.8%, Rt = 1.654 min; MS calculated 15 Ei: 1456.7; MS measured: 1457.8 [M+H]*.'H NMR (400 MHz, DMSO-ds) 5 8.52 (t, J= 6.7 Hz, 1H), 8.37 (t, J = 6.0 Hz, 1H), 8.19 — 8.12 (m, 2H), 8.02 (t, J= 5.7 Hz, 1H), 7.93 (t, J= 8.6 Hz, 1H), 7.80 — 7.75 (m, 2H), 7.28 — 7.24 (m, 6H), 7.00 (s,2H), 6.51 (br, 1H), 5.48 (s, 2H), 5.42 (s, 2H), 4.61 — 4.60 (m, 2H), 4.51 (q, J = 8.3, 7.8 Hz, 1H), 3.77 — 3.69 (m, 6H), 3.61 — 3.59 (m, 6H), 3.49 — 3.47 (m, 28H), 3.28 — 3.22 (m, 6H), 3.18 — 3.16 (m, 2H), 3.09 — 3.08 (m, 1H), 3.06 — 3.05 (m, 1H), 2.49 (s, 20 3H), 2.39 (t, J= 6.6 Hz, 2H), 2.03 (t, J = 7.3 Hz, 2H), 1.88 — 1.82 (m, 2H), 1.58 — 1.52 (m, 1H), 1.48 — 1.42 (m, 4H), 1.25 — 1.17 (m, 6H), 0.88 (t, = 6.0 Hz, 3H). Intermediate compound F1014-01 was prepared as follows: OH 6 fe) o 4 H + total o PPTS no ZA Sm fe Prey ho . SPN OME. 98%. N 3 vn / nc 2 F HO 20 F mes F1014-01 GOR: Compound STI-F14 (170 mg, 0.34 mmol). 25b (151 mg, 0.41 mmol) #ll Xf} FA EGR 25 We FRO mg was added to the reaction flask, DMF (10.0 mL) was added, BAUER FH 95°C RIVA 3 hours later stopped 90 WO 2024 / 255740 PCT / CN2024 / 098491 TE BRD EAS RRR aR EFDA BT 14-4 (DCM: MeOH=50: 1), 180 mg 4447 F1014-01 was obtained as a yellow solid, yield 63.8%. LCMS (254 nm purity 51.39%, Rt = 1.992 min; MS ++ iA: 801.3; MS measured value: 802.3 [M+H]". 5 ” Example 38: Synthesis of drug linker conjugate STLF101s HOO—, Q ma or H + NH O PPTS ele as mee yaNO IQ DMF, 95°C, 3h NYA HOT 一 9 9 HaC 分 必 Pb N 25b F | Ho = _9 H3C STLF15 F F1015-01 1)Piperidine, DMF 5 NN rs yD : \ j O N STI-F1015 H3C ! Step 1: Compound STI-F1S (390 mg, 0.77 mmol), 25b (339 mg, 0.92 mmol) and pyridinium p-toluenesulfonate (20 mg) were added into a reaction flask, DMF (12.0 mL) was added, and the mixture reacted at 95°C for 3 hours, then the reaction was stopped. The system was concentrated to dryness under reduced pressure and purified by column chromatography with DCM:MeOH=50:1, to obtain 180 mg of compound FE1015-01 as a yellow solid, with a yield of 18.5%. LCMS (254 nm, purity 47%, Rt = 1.891 min; MS calculated value: 815.3; MS found value: 816.4 [M+H]+. Step 2: Compound F1015-01 (47%, 180 mg, 0.11 mmol) was dissolved in DMF (2.7 mL), piperidine (0.3 mL) was added, and the reaction was carried out at room temperature for 30 minutes. The system was concentrated to dryness, slurried with acetone, filtered by suction. The filter cake and HATU (125 mg, 0.30 mmol) were added into a reaction flask, under nitrogen protection, DMF (3.0 mL) and Example 37 F1006-S3 (107 mg, 0.12 mmol) and DIPEA (43 mg, 0.30 mmol) were added, and the reaction was carried out at room temperature for 0.5 hours. LCMS monitored that the reaction was complete, then the reaction was stopped. The system was concentrated to dryness and subjected to medium-high pressure reverse-phase preparation (acid mobile phase), after lyophilization, 15 mg of compound STI-F1015 was obtained as a light yellow solid, with a yield of 9.3%. LCMS (254 nm, purity 97.56%, Rt= 1.655 min; MS calculated value: 1470.7; MS found value: 1469.6 [M-H]. ¹H NMR (400 MHz, DMSO-d₆ δ 8.52-8.47(m, 2H), 8.31 (t, J= 5.7 Hz, 1H), 8.17 (t, J= 5.7 Hz, 1H), 8.12 (d, = 8.1 Hz, 1H), 8.02 (t, J=5.8 Hz, 1H), 7.81 (t, J= 5.7 Hz, 1H), 20 7.60 (d, J= 10.7 Hz, 1H), 7.37 (s, 1H),7.28 — 7.20 (m, 5H), 7.00 (s, 2H), 6.54 (br, 1H), 5.42 (s, 2H), 5.38 (s, 2H), 4.57 — 4.55 (m, 2H), 4.53 — 4.48 (m, 1H), 3.78 — 3.69 (m, 7H), 3.63 — 3.58 (m, 7H), 3.49 — 3.47 (m, 28H), 3.29 (d, J = 6.1 Hz, 2H), 3.19 — 3.14 (m, 2H), 3.09 — 3.08 (m, 1H), 3.06 — 3.05 (m, 1H), 2.45 (s, 3H), 2.39 (t, J = 6.6 Hz, 2H), 2.07 — 1.97 (t, J= 7.3 Hz, 4H), 1.88 — 1.82 (m, 2H), 1.64 — 1.54 (m, 2H), 1.50 — 1.42 (m, 5H), 1.21 — 1.15 (m, 6H), 0.88 (t, J = 6.0 Hz, 3H). 25 Example 39: Synthesis of drug linker conjugate STI-F1018 91 WO 2024 / 255740 PCT / CN2024 / 098491 H 9 H BAN : 2) F1018-01, DIEA, DMF By : — atFaDeM 0 fe) ss Nm oH, caer O fe] N Step 1: 4 F1005-01 (65 mg, 0.24 mmol) deep in dioxane (2.0 mL) +, BRE, then add NHS (34 mg, 0.29 mmol) and DCC (60 mg, 0.29 mmol), YHRISRDY 1 hour, TLC After the reaction was completed, the reaction solution was filtered, and the filter cake was rinsed with DCM (5 mL * 3). The filtrate was concentrated to dryness at room temperature and used directly for the next step.The crude product and 5 F1018-01 (145 mg, 0.24 mmol) were dissolved in DMF (2 mL), JA DIEA (130 pL, 0.72 mmol / L), and stirred in an ice bath for 2 hours. After the reaction was monitored by LCMS, acetic acid was added to quench the reaction until the pH was acidic. The mixture was concentrated at 455°C using an oil pump. The crude product was then prepared by medium-low pressure reverse phase reaction (mobile phase: acetic acid / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 200 mg of compound F1018-02 was obtained, which was a transparent oil with a yield of 96.9%. LCMS (254 nm) purity was 92.1%, Rt = 1.95 min; MS calculated value: 849.3; MS measured value {41:850.5 [M+H]*. 10 Step 2: K F1018-02 (53 mg, 0.06 mmol) Dissolved in dichloromethane (2.0 mD), under nitrogen protection, NHS (9.0 mg, 0.07 mmol) and DCC (16 mg, 0.07 mmol) were added. The reaction was monitored by FIBRIN and TLC. After the reaction was complete, the reaction solution was filtered, and the filter cake was washed with DCM (5 mL * 3). The filtrate was concentrated to dryness at room temperature and used directly for the next step. The above concentrate was dissolved in DMF (2.0 mD), and F1016-06 (43 mg, 0.05 mmol) and DIEA (49 mg, 0.37 mmol) were added. The reaction was incubated on ice for 2 hours. The reaction was monitored by LCMS until complete. The oil pump was stopped at 4°C. The crude product was purified by column chromatography (dichloromethane / methanol = 10 / 3) to obtain 15-46 mg of crude compound. The crude compound was dissolved in 10% TFA / DCM (2 mD). Under nitrogen protection, the mixture was stirred in an ice bath for 1 hour, and the reaction was monitored by LCMS to ensure completion. The reaction solution was adjusted to pH 7-8 with triethylamine, dried under nitrogen, and then prepared by high pressure reverse phase (mobile phase: Z58 / 0.05% = CBR IK IAD). After lyophilization, 5.69 mg of compound STI-F1018 was obtained as a yellow solid. LCMS (254 nm) showed a purity of 93.5%, Rt = 1.57 min; MS calculations...Value: 1602.7; MS found: 1603.6 [M+H]+. ¹H NMR (400 MHz, DMSO-d₆) δ 8.95 (s, 2H), 8.32 (t, J= 6.0 Hz, 1H), 8.36 - 8.25 (m, 2H), 8.10 (d, J = 8.0 Hz, 1H), 7.99 (t, J= 6.0 Hz, 1H), 7.87 (s, 2H), 7.61-7.58 (m, 3H), 7.31 (d, J = 2.0 Hz, 1H), 7.21 - 7.08 (m, 5H), 6.53 (brs, 1H), 5.41 (d, J = 16.8 Hz, 4H), 4.59 - 4.54 (m, 3H), 4.24 - 4.23 (m, 1H), 3.88 - 3.79 (m, 3H), 3.67-3.59 (m, 2H), 3.57 (t, J= 6.8 Hz, 3H), 3.47-3.42 (m, 4H), 3.28-3.23 (m, 4H), 3.16 - 3.12 (m, 6H), 2.86 - 2.74 (m, 4H), 2.55 (t, J = 7.2 Hz, 2H), 2.32 - 2.22 (m, 5H), 1.88 - 1.78 (m, 4H), 1.77 - 1.62 (m, 5H), 1.57 - 1.50 (m, 3H), 1.34 - 1.27 (m, 2H), 0.86 (t, J = 7.6 Hz, 3H). Intermediate compound F1018-01 is prepared as follows: 92 WO 2024 / 255740 PCT / CN2024 / 098491 HCl. FmocHN- FmocNH _F 1018-04-02 NHBoc HCl / Dioxane FmocHN OH HATU, DIEA F1018-01-01 F1018-01-03 F1018-01-04 Piperidine / DMF F1018-01-06 F1018-01 Step 1: F1018-01-01 (541 mg, 1.0 mmol) is dissolved in dioxaneIn (10.0 mD), under gas protection, HATU (380 mg, 1.0 mmol) and DIEA (258 mg, 2.0 mmol) were added. The mixture was stirred at room temperature for 5 minutes, then F1018-01-02 (318 mg, 1.0 mmol, HCl salt) was added. After the reaction was monitored by LCMS, the reaction solution was concentrated at 4°C to obtain the crude product. The product was then prepared by medium-low pressure reverse phase reaction (mobile phase: acetic acid / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 680 mg of compound F1018-01-03 was obtained as a transparent oil, with a yield of 84.4%. LCMS (254 nm) showed a purity of 94.5%, Rt = 1.89 min; MS calculated value: 805.4; MS measured value: 806.5 [M+H]*. Step 2: F1018-01-03 (200 mg, 0.24 mmol) was dissolved in 4M HCVEA (4.0 mD) and reacted at room temperature for 1.0 h. After the reaction was monitored by TLC, the reaction solution was concentrated at 4°C, and some EA was removed with petroleum ether (5 mL * 3) to obtain 174 mg of F1018-01-04, which was directly used for the next step. Step 3: F1018-01-04 (174 mg, crude) was dissolved in DMF, and F1018-01-05 (143 mg, 1.24 mmol) and DIEA (160 mg, 1.24 mmolD) were added. The mixture was stirred at room temperature for 0.5 h, and reacted with LCMS HS WIR SEE, TA ZR until the pH reached acidity. The mixture was concentrated at 4°C by oil pump and prepared by medium-low pressure reverse phase (mobile phase: Z.H8 / 0.05% = Ft). CRIA, after lyophilization, yielded 180 mg of compound F1018-01-06 as a transparent oil, with a yield of 88.4%. LCMS (254 nm) purity 15 93.1%, Rt = 1.95 min; MS calculated value: 821.3; MS measured value: 822.5 [M+HJ]. Step 4: Compound F1018-01-06 (180 mg, 0.22 mmol) was dissolved in DMF (2.0 mL), JA 200 nh WWE, BURP TEE BY 0.5 h.The reaction was monitored by LCMS until it was complete. After concentrating the reaction solution to dryness using a 4% C oil pump, petroleum ether was added and the mixture was stirred (2 mL * 3). The supernatant was discarded to obtain 145 mg of compound F1018-01. The crude product was used directly for the next step. The intermediate compound F1016-06 was prepared as follows: en 1) DCC, NHS, DCM aN 1) DCC, NHS, THF ee Pb(OAc),, Pyridine meters 20 F1018-05 F1016-06 Step 1: 4% F1016-01 (1.0 g, 2.58 mmol) was dissolved in dioxane (20.0 mL), under nitrogen protection, and then NHS (357 mg, 3.1 mmol) and DCC (640 mg, 3.1 mmol) were added. The reaction was carried out in an ice bath for 1 hour. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with DCM (3) The filtrate was concentrated to dryness at room temperature, and the crude product was used directly for the next step; the above crude product was reacted with NH2-Lys(Boc)-OH (635 mg, 2.58 mmol / L) and NaHCO3 (665 mg, 7.75 mmol / L), and the reaction was carried out in an ice bath for 2 hours. The reaction was detected by LCMS. 1M hydrochloric acid was added to adjust the pH to 1~2. Ethyl acetate was taken (10 mL x 3), washed with saturated brine (10 mL x 2), dried with anhydrous Na2SO4, concentrated, and the crude product was prepared by medium-low pressure reverse phase (mobile phase: ethyl acetate / 0.05% trifluoroacetic acid aqueous solution). After drying, 900 mg of crude product was obtained. Compound F1016-02, a white pentoxide, yield 6.7%. LCMS (254 nm) showed a purity of 95.1%, Rt = 2.12 min; calculated MS value: 615.2; measured MS value: 616.3 [M+H]+. Step 2: F1016-02 (900 mg, 1.46 mmol) was placed in AF YU ARKIN (10.0 mD) under nitrogen protection, and then NHS (168 mg, 1.46 mD) was added.mmol and DCC (300 mg, 1.46 mmol), FIIS RAL for 1 hour, TLC monitoring of the reaction completion RUA, the reaction solution was filtered, the filter cake was washed with THF (5 mL*3), the mother liquor was concentrated and used directly for the next step, 1.1 g, crude product; 10 "The above concentrate was dissolved in DMFVH2O / acetone=1:1:1 (10 mD), glycine (110 mg, 1.46 mmolD) and NaHCO; (369 mg, 4.38 mmol), the reaction was carried out in an ice bath for 2 hours, the reaction was detected by LCMS, 1N hydrochloric acid was added to make the pH=1-2, ethyl acetate was taken (10 mL*#*3), washed with saturated brine (10 mL*2), dried with anhydrous Na2SO4, concentrated, the crude product was prepared by medium and low pressure reverse phase (mobile phase, acetonitrile / 0.05%). Trichloroacetic acid aqueous solution was lyophilized to give 800 mg of compound F1016-03, a white solid, with a yield of 81.5%. LCMS (254 nm, purity 96.1%, Rt = 2.02 min; MS calculated value: 672.3; 15-MS measured value: 673.4 [M+H]'). Step 3: F1016-03 (800 mg, 1.19 mmol iF) was added to 16 mL of PUSKIRA, 4 mL of aH, IA Pb(OAc)4 (633 mg, 1.42 mmol OD) and pyridine (113 mg, 1.42 mmol OD), and BAUR. 85°C CHPE for 2.5 hours. The raw material was completely consumed by LCMS. The mixture was concentrated to dryness at 45°C. The crude product was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 3) to give 460 mg Compound F1016-04 is a white solid with a yield of 56.3%. LC-MS (254 nm, purity 93.2%, Rt = 1.85 min) showed calculated values ​​of 686.3 and measured values ​​of 704.1 [M+HNH4⁺]. ¹H NMR (400 MHz, DMSO-ds) showed 8.91 (t, J = 6.8 Hz, 1H), 8.10 (dJ = 8.0 Hz, 1H), and 7.87 (d, J = 7.2 Hz, 2H).7.63 - 7.58 (m, 3H), 7.42-7.38 (m, 2H), 7.32-7.26 (m, 7H), 6.72 (t, J = 4.8 Hz, 1H), 5.11-5.06 (m, 2H), 4.31 - 4.13 (m, 5H), 3.03-2.73 (m, 4H), 1.98 (s, 3H), 1.63-1.49 (m, 2H), 1.34-1.18 (m, 13H). Step 4: Under nitrogen protection, add 25 A STI-F6 (120 mg, 0.25 mmol) 41 ml PPTS to compound F1016-04 (176 mg, 0.25 mmol) F DMF (5 mD). (12 mg, cat), RY at 95°C for 2 hours. The reaction solution was concentrated to dryness under reduced pressure at 45°C. The crude product was purified by column chromatography (dichloromethane / methanol = 30 / 1) to give 50 mg of compound F1016-05 as a yellow solid with a purity of 50%; MS calculated value: 1093.6; MS measured value: 1094.6 [M+HT']. Step S: Compound F1016-05 (60 mg, 0.05 mmol) was dissolved in DMF (2.0 mD), 200 pL piperidine was added, and the reaction was stirred under choke protection for 0.5 hours. The reaction was monitored by LCMS to ensure complete reaction. The reaction solution was concentrated to dryness using a 4% C oil pump. 30% petroleum ether was added and the mixture was pulped (2 mL x 3). The supernatant was discarded, and the solution was concentrated again to obtain 43 mg of compound F1016-06. The crude product was used directly for the next step. Example 40: Synthesis of drug linker conjugate STI-F1019 5, 'a te human pperdine OME a8, 5, : A human ° Compound F1013-05 (30 mg, 0.03 mmol / D) from Example 36 was dissolved in DMF (2.0 mD), 200 pL of 3, " Piperamide was added, and the reaction was stirred under nitrogen protection for 0.5 hours. The reaction was monitored by LCMS to ensure complete reaction. The reaction solution was concentrated to dryness by oil pump at 45°C and then 94 WO 2024 / 255740 PCT / CN2024 / 098491 petroleum ether was added and slurryed (2mL*3), felt LYK, FKURARTS I 24 mg crude product without Fmoc. F1018-02 (32 mg, 0.03 mmol) from Example 39 was dissolved in DMF (2.0 mD) under nitrogen protection. Then, HATU (22 mg, 0.05 mmol) and DIEA (15 mg, 0.11 mmol) were added, followed by PES for 1 minute. The crude Fmoc-free product was then added, and the reaction was continued for 1 hour. After the reaction was complete as monitored by LCMS, the reaction solution was concentrated to dryness at 4°C. The crude product was purified by column chromatography (5% dioxane / 20% methanol) to obtain the condensation product. The condensation product was dissolved in 10% TFA / DCM (2 mD), and stirred in an ice bath under nitrogen protection for 1 hour. The reaction was monitored by LCMS to ensure completeness. The reaction solution was adjusted to pH 7-8 with triethylamine hexamethonium, dried under pneumatic pressure, and then prepared by high-pressure reverse-phase chromatography (mobile phase: ZH2O / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 2.87 mg of compound STILF1019 was obtained as a white solid. LCMS (254 nm, purity 99.6%, Rt = 1.51 min; MS calculated value: 1455.6; MS measured value: 1456.7) was performed. ¹H NMR (400 MHz, 10 DMSO-ds) showed the following values: 9.11 (s, 2H), 8.78 (t, J = 6.4 Hz, 1H), 8.41 - 8.39 (d, J = 7.6 Hz, 1H), 8.11 (d, J = 8.0 Hz, 1H), 8.03 (t, J = 4.2 Hz, 1H), 7.88 (s, 2H), 7.65-7.59 (m, 3H), 7.39 (s, 2H). 1H), 6.57 (brs, 1H), 5.48 (d, J = 38 Hz, 4H), 4.59 - 4.57 (m, 2H), 4.29 - 4.26(m, 1H), 4.01 - 3.90 (m, 3H), 3.70 - 3.59 (m, 7H), 3.45 - 3.30 (m, 30H), 3.32 - 3.16 (m, 4H), 3.09 - 3.07 (m, 3H), 2.77 - 2.73 (m, 2H),2.63-2.53 (m, 4H), 2.44 (s, 3H), 2.32 - 2.22 (m, 5H), 1.89 - 1.64 (m, 12H), 1.30 - 1.23 (m, 2H), 0.87 (t, J = 7.2 Hz, 15 3H). Example 41: Synthesis of drug linker conjugate STI-G1001 Fioos-ss.HAru DIPEA, DMF ec An 4, ra Ag FN~ \ , Step 1, compound STI-G$ (140 mg, 0.275 mmolD) was added to 2.8 mL of NN-dimethylacetamide, 20 A 25b (152 mg, 0.42 mmolD) and PPTS (7 mg, 0.0275 mmolD) were added, Under nitrogen protection, the reaction was carried out at 80°C for 4 hours. After the reaction was completed, the reaction solution was evaporated, acetone was added and the mixture was stirred, filtered, and the filtrate was evaporated and purified by column chromatography to obtain compound G1001-01 (100 mg, 0.122 mmol) as a yellow solid, with a yield of about 40%. LCMS (254 nm) Rt = 1.918 min; MS calculated value: 817.3; MS measured value: 816.3 [MH]-. Step 2: Compound G1001-01 (100 mg, 0.122 mmol) was added to 2 mL of 10% piperidine / 25" dioxane solution and reacted at room temperature for 30 minutes. After the reaction was completed, the reaction solution was evaporated, the product was stirred with acetone, filtered, and the filter cake product was G1001-02, about 50 mg, as a yellow solid. The yield was approximately 68%. LCMS (254 nm) Rt = 1.539 min; MS calculated value: 595.24; MS measured value: 596.30 [M+H]. Step 3: Compound G1001-02 (50 mg, 0.084 mmol) was placed in a reaction flask, and F1006-S3 (75 mg, 0.084 mmol) from Example 37 was added. 1 mL of DMF was added to allow it to dissolve, followed by the addition of DIPEA (30 μL, 0.168 mmol) and HATU (60 mg, 0.168 mmol).mmolD, at room temperature for 30 minutes. After the reaction was monitored by LCMS, the reaction solution was evaporated to dryness, and then separated by high pressure (mobile phase: acetic acid / 0.05% formic acid aqueous solution). After lyophilization, approximately 15 mg of compound STI-G1001 was obtained as a yellow solid. LCMS (254 nm, purity 97.5%, Rt = 1.620 min; MS calculated value: 1472.68; MS measured value: 737.50) 95 WO 2024 / 255740 PCT / CN2024 / 098491 [M / 2+H]*. 'H NMR (400 MHz, DMSO-ds) 6 9.78 (s, 1H), 9.70 (s, LH), 8.95 (d, J = 8.4 Hz, 1H), 8.87 (d, J = 8.4 Hz, 1H), 8.72 (t, J= 6.8 Hz, 1H), 8.33 (t, J= 5.6 Hz, 1H), 8.18 - 8.11 (m, 2H), 8.04-7.99 (m, 2H), 7.81 - 7.99 (m, 1H), 7.30 (s, 2H), 7.22 (s, 2H), 6.99 (s, 1H), 6.52 (brs, 1H), 5.43 (s, 2H), 5.29 (s, 2H), 4.74 (d, J= 6.8 Hz, 1H), 4.51 - 4.47 (m, 1H), 4.22 (s, 2H), 4.15 (s, 2H), 3.80 - 5 3.76(m, 2H), 3.72 - 3.67 (m, 2H), 3.64 - 3.57 (m, 2H), 3.49 (s, 34 H), 3.24 - 3.15 (m, 2H), 3.14 - 3.10 (m, 2H), 3.05 - 3.01 (m, 2H), 2.38 (t, J= 6.4 Hz, 2H), 2.04 - 1.99 (m, 2H), 1.90 - 1.81 (m, 2H), 1.73 (s, 2H), 1.47 - 1.44 (m, 4H), 1.40 - 1.36 (m, 2H), 1.23(s, 4H), 0.91 - 0.87 (m, 6H). Example 42: Synthesis of drug linker conjugate STI-F1031 er ne yh yy hn yt SL,eon O © 4 N, , , NH: oO oO. O 2 0 © o of F1031-S3 O 0 oto x O o thor, 10 al met Step 1: Compound F1031-01 (2.88 g, 12.0 mmol) was added to 4 F Z.f5 (48.0 mL), followed by addition of sodium carbonate (1272 mg, 12.0 mmol), under nitrogen protection. tert-butyl bromoacetate (585 mg, 3.0 mmol) was added dropwise to the system, followed by reaction at room temperature for 24 hours. The reaction was stopped, filtered under suction, the filtrate was concentrated under reduced pressure, extracted with ethyl acetate and water, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (PE:EA=1:10) to obtain 1.1 g of compound F1031-02 as a yellow solid, with a yield of 25.9%. 15 LCMS (254 nm) purity 93.2%, Rt = 1.816 min; MS calculated value: 354.2; MS found value: 355.3 [M+H]+. Step 2: Compound F1031-02 (1200 mg, 3.39 mmol) was added to F Zi (20.0 mL), followed by addition of sodium carbonate (1030 mg, 10.0 mmol) and AUR. TBR RAIN Z RASC (214 mg, 0.5.3 mmol) in AY ZA, and the reaction was allowed to proceed at room temperature for 1 hour. The reaction was stopped, filtered under suction, the filtrate was concentrated under reduced pressure, and purified by column chromatography (PE:EA=30:1) to obtain 1.5 g of compound F1031-01b-1 as a yellow oily liquid, with a yield of 88.2%. LCMS (254 nm) purity 93.2%, Rt = 2.421 min; 20 MS calculated value: 302.2; MS found value: 503.6 [M+H]+. Step 3: Compound F1031-01b-1 (1.0 g, 2.0 mmol) was added to HF FR (20.0 mL), followed by addition of 10% Pd / C (100 mg), replaced with hydrogen, and reacted overnight at room temperature. The reaction was monitored by LCMS to be complete, then the reaction was stopped, filtered under suction, and the filtrate was concentrated to dryness under reduced pressure to obtain 445 mg of compound F1031-02b, which is anA colored oily liquid, yield 95.9%. LCMS (254 nm) purity 91.0%, Rt = 1.910 min; MS calculated value: 232.1; MS measured value: 233.2 [M+H]°. 25 Step 4: 4,7,10,13,16,19,22.25-octaoxahexacosanoic acid (412 mg, 1.0 mmol) XH LANA (221 mg, 1.2 mmol AAW APRI (5.0 mL), ATS, B22 0~$C, add NN'-dicyclohexylcarbodiimide (247 mg, 1.2 mmol), react at room temperature for 4 hours, filter, concentrate the filtrate to dryness under reduced pressure, add compound F1031-02b (126 mg, 0.5 mmol) DMF (5.0 mL), under vibratory protection, Add DIPEA (258 mg, 2.0 mmol / L), react overnight at room temperature, monitor the reaction completion by LCMS, concentrate the system to dryness under reduced pressure, and use the crude product (F1031-03bp) directly in the next step. 96 WO 2024 / 255740 PCT / CN2024 / 098491 Step $: Redissolve the crude product from the previous step in DMF (5.0 mL), JIA F1031-S3, HATU (570 mg, 1.5 mmol) & DIPEA (387 mg, 3.0 mmol), and under nitrogen protection, react at room temperature for 1 hour. Monitor the reaction completion by LCMS, stop the reaction, concentrate the system to dryness, and prepare by reverse phase (mobile phase: acetonitrile / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 240 mg of compound F1031-04b is obtained, which is a colorless oily liquid. Yield 40.2%. LCMS (254 nm), purity 94.1%, Rt = 1.526 min; MS 5" calculated value: 1184.7; MS measured value: 1183.35 [MH]. Step 6: Compound F1031-04b (100 mg, 0.084 mmol AF) was added to a 25% trifluoroacetic acid solution in dioxane. The reaction was monitored to be complete by LCMS at 27 nm. The solution was concentrated to dryness under reduced pressure to obtain 130 mg.Compound F1031-08 was a colorless oily liquid with a yield of 100.0%. LCMS (254 nm, purity 95.1%, Rt = 1.513 min; MS calculated value: 1128.6; MS measured value: 1127.7 [M-HJ]-). Step 7: Compound F1010-03 (40 mg, 0.045 mmol YF DMF (0.9 mL) + A from Example 33 was added to piperidine (0.1 mL), reacted at room temperature for 20 minutes, concentrated to dryness, washed with PE, and dried under reduced pressure. DME (2.0 mL), HATU (34 mg, 0.09 mmol), F1031-08 (56 mg, 0.05 mmol), and triethylcarbamide (23 mg, 0.23 mmol) were added to the residue. AER, FEW 1.0 h, stop the reaction, concentrate the system to dryness, medium and high pressure Bert tl GRA: 2.8 / 0.05% = ZR), YRS BI 17 mg Compound STI-F1031, is a pale yellow 15" solid, yield 21.3%. LCMS (254 nm) purity 98.58%, Rt = 1.623 min; MS calculated value: 1776.9; MS measured value: 1776.0 [MH]. 'H NMR (400 MHz, DMSO-ds) & 8.63 — 8.56 (m, 1H), 8.48 — 8.35 (m, 2H), 8.15 (t, J= 6.8 Hz, 1H), 8.10 — 8.04 (m, 1H), 8.00 (d, J = 6.5 Hz, 1H), 7.60 (d, J= 10.6, 1H), 7.37 (s, 1H) , 6.98 (s, 2H), 6.55 (br, 1H), 5.48 (s, 2H), 5.43 (s, 2H), 4.72 — 4.69 (m, 1H), 4.56 — 4.46 (m, 2H), 4.25 — 4.18 (m, 3H), 4.12 (t, J= 4.6 Hz, 1H), 4.08 — 3.96 (m, 2H), 3.70 — 3.65 (m, 3H), 3.62 — 20 3.58 (m, 4H), 3.51 —3.46 (m, 63H), 3.23 (s, 6H), 3.05 — 2.99 (m, 4H), 2.61 — 2.56 (m, 4H), 2.44 (s, 3H), 2.04 — 1.97 (m, 2H), 1.88 — 1.83 (m, 2H), 1.74 (q, J= 7.4 Hz, 2H), 1.64 (q, J = 6.8 Hz, 2H), 1.51 — 1.42 (m, 2H), 1.21 (d, J= 7.1 Hz, 3H), 0.87 (t, J= 7.2 Hz, 3H), 0.82 (dd, J= 11.2, 4.9 Hz, 6H). The intermediate compound F1031-S3 was prepared as follows: [The rest of the text appears to be a series of symbols and characters, possibly a corrupted or incomplete sample. A direct translation is not possible without further context or clarification.] Column chromatography purification (DCM:EA=20:D) yielded 1.1 g of compound F1031-S2 as a white solid with a yield of 30 3K 5.7%. LCMS (254 nm) purity 93.2%, Rt = 1.723 min; MS calculated value: 282.2; MS measured value: 283.1 [MT+HJ]-. Step 2: Compound F1031-S2 (423 mg, 1.5 mmol JAF 2% trichloroacetic acid in dioxane solution) was stirred at room temperature for about 2 hours. The reaction was monitored by LCMS until complete. The solution was concentrated to dryness under reduced pressure to obtain F1031-S3, which was directly used in the next step. 35” Example 43: Synthesis of drug linker conjugate STI-F1034 97 WO2024 / 255740 PCT / CN2024 / 098491 9 NM SY 2 H 9 nN \-¥ 2 Step 1: Compound STI-F14 (276 mg, 0.56 mmol D, Compound F1010-02 (269 mg, 0.56 mmol) from Example 33) was reacted with PPTS (20 mg) JIA RIA, DOA DMF (10 mL), BURP, FPL 90*C for 5 hours. The reaction was stopped, and the system was concentrated to dryness under reduced pressure and purified by column chromatography (DCM:MeOH=100:1~60:D) to obtain 154 mg of F1033-01, AYER IA, with a yield of 30.0%. LCMS (254 nm) calculated value: purity 76.8%, Rt = 2.043 min; MS calculated value: 914.4; MS measured value: 915.4 [M+H]. Step 2: Compound F1033-01 (154 mg, 0.17 mmol / D) was added to 3 mL of DMF, and 300 uh ORME was added. The mixture was stirred under atmospheric protection for 0.5 hours, and the reaction was monitored by LCMS to ensure complete reaction. The reaction solution was concentrated to dryness by a 4°C oil pump, and then petroleum ether was added for slurrying. After filtration, 100 mg of compound F1033-02 was obtained as a pale yellow solid with a yield of 85.0%. LCMS (254 nm) purity 10 98.7%, Rt = 1.692 min; MS calculated value: 692.3; MS measured value: 693.4 [M+H]. 'H NMR (400 MHz, DMSO-ds) 8 8.74 (t, J = 6.6 Hz, 1H), 8.13 (s, 1H), 7.91 (d, J = 8.4 Hz, 1H), 7.76 (d, J = 10.7 Hz, 1H), 7.27 (s, 1H), 6.51 (s, 1H), 5.46 (s, 2H), 5.43 (s, 2H), 4.67 — 4.55 (m, 2H), 4.37 — 4.29 (m, 1H), 3.63 (d, J= 11.2 Hz, 2H), 3.44 — 3.37 (m, 4H), 3.07 — 3.01 (m, 1H), 2.48 (s, 3H), 1.98 — 1.88 (m, 2H),1.89 - 1.79 (m, 4H), 1.60 — 1.46 (m, 2H), 1.26 (d, J = 7.0 Hz, 3H), 1.24 — 1.14 (m, 2H), 0.90 — 15 0.86 (m, 6H), 0.79 (d, J = 6.8 Hz, 3H). Step 3: F1033-02 (67 mg, 0.097 mmol) and F1031-08 (120 mg, 0.106 mmol) from Example 42, plus HATU (74 mg, 0.194 mmol), were bathed in 3 mL DMF, JA DIPEA (82 pL, 0.485 mmol), #4 protection, and reacted at room temperature. ZH / 0.05% = HZ Bek YO), UF Ja Gl 12.5 mg Compound STI-F1034, is a yellow oily substance with a yield of 6.9%. LCMS (254 nm) 20 1 RE 100%, Rt = 1.684 min; MS calculated value: 1802.9; MS measured value: 1803.7 [M+HT ]. H NMR (400 MHz, DMSO-ds) 5 8.47 (s, 1H), 8.25 — 8.17 (m, 1H), 8.13 — 7.99 (m, 2H), 7.92 (d, J= 8.5 Hz, 1H), 7.77 (d, J = 10.8 Hz, 1H), 7.28 (s, 1H), 6.99 (s, 2H), 6.50 (s, 1H), 5.48 (s, 2H), 5.42 (s, 2H), 4.66 — 4.50 (m, 7H), 4.32 — 4.19 (m, 2H), 3.67 — 3.44 (m, 66H), 3.23 (s, 6H), 3.09 — 2.93 (m, 3H), 2.70 — 2.57 (m, 4H), 2.48 (s, 3H), 2.44 — 2.36 (m, 4H), 2.36 — 2.30 (m, 1H), 2.05 — 1.94 (m, 2H), 1.93 — 1.78 (m, 25 5H), 1.44 — 1.34 (m, 2H), 1.31 — 1.21 (m, 5H), 1.23 — 1.14 (m, 2H), 0.93 — 0.80 (m,9H). Example 44: Synthesis of drug linker conjugate STILF1035 F810 8 AL Ag}, Oo ALA, "OFF N Step 1: 4,7,10,13,16,19,22.25- Octoxohexadecanoic acid (412 mg, 1.0 mmol) XA HANA (221 mg, 30 12 mmol) JU AVUAPAMA (5.00 mL), AUS, BEER 0~$C, added N,N ORO IK 98 WO 2024 / 255740 PCT / CN2024 / 098491 (247 mg, 1.2 mmol), reacted at room temperature for 4 hours, filtered, HERO RRA, added compound F1031-02b (100 mg, 0.43 mmol) J¢ DMF (3.0 The mixture was choked under suffocation, and then DIPEA (111 mg, 0.86 mmol D) was added. The reaction was carried out overnight at room temperature. The reaction was monitored by LCMS until completion. The system was concentrated to dryness under reduced pressure, and the residue was redissolved in DME (5.0 mL). F1035-S4, HATU (327 mg, 0.86 mmol) and DIPEA (166 mg, 1.29 mmol) were added. The reaction was carried out for 1 hour. LCMS showed that compound F103$-01 was a colorless oily liquid with a yield of 33.5%. The purity was 98.0%, Rt = 1.537 min; MS calculated value: 1241.6; MS measured value: 1240.6 [MH]-. Step 2: Compound F1035-01 (59 mg, 0.05 mmol JAF 25% trichloroacetic acid in dichloromethane solution) was stirred at room temperature for about 5 hours. The reaction was monitored by LCMS until complete. The solution was concentrated to dryness under reduced pressure to obtain the deprotected product 103S-01. Compound F1010-03 (40 mg, 0.045 mmol D) from 10 SEH) 33 was added to DMF (0.9 mL D), and piperidine (0.1) was added.The reaction was carried out at room temperature for 20 minutes, the system was concentrated to dryness, the residue was washed with petroleum acid, and dried under reduced pressure. F1035-01 deprotected product, DMF (2.0 mL), HATU (34 mg, 0.09 mmol D), and triethylamine (23 mg, 0.23 mmol), A Und. FSR were added to the residue. After 0.5 hours, the reaction was stopped, the system was concentrated to dryness, and prepared by high-pressure reverse phase (BAA: ethyl acetate / 0.05% trifluoroacetic acid aqueous solution). After lyophilization, 20 mg of compound STI-F1035 was obtained as a yellow solid with a yield of 15 38 24.2%. LCMS (254 nm) showed a purity of 95.23%, Rt = 1.633 min; MS calculated value: 1833.9; MS measured value: 1833.0 [MH]. 'H NMR (400 MHz, DMSO-ds) 5 9.10 (s, 2H), 8.60 (t, J = 6.0 Hz, 1H), 8.45 — 8.40 (m, 2H), 8.16 (t, J = 6.7 Hz, 1H), 8.08 (d, J = 8.5 Hz, 1H), 7.91 — 7.80 (m, 1H), 7.58 (d, J = 10.5 Hz, 1H), 7.41 (s, 1H), 6.58 (br, 1H), 5.47 (s, 2H), 5.44 (s, 2H), 4.56 — 4.49 (m, 2H), 4.26 — 4.18 (m, 3H), 4.13 — 4.11 (m, 1H), 4.05 - 3.95 (m, 2H), 3.73 - 3.68 (m, 2H), 3.61 - 3.56 (m, 4H), 3.51 - 3.46 (m, 20 65H), 3.40 (s, 6H), 3.23 (s, 3H), 2.63 - 2.55 (m, 4H), 2.43 (s, 3H), 2.41 — 2.39 (m, 2H), 2.01 — 1.91 (m, 2H), 1.90 — 1.83 (m, 2H), 1.76 — 1.73 (m, 2H), 1.68 — 1.60 (m, 2H), 1.23 (d, J= 4.6 Hz, 3H), 0.87 (t, J = 7.4 Hz,3H), 0.82 (dd, J= 12.7, 6.5 Hz, 6H). The intermediate compound F1035-S4 was prepared as follows; Step 1: J F1035-S1 (303 mg, 1.5 mmoloD), N-Boc-4-pentadienylamine (330 mg, 1.8 mmoloD), uranium iodide (30 mg, 0.3 mmoloD) were added to the reaction flask, and under nitrogen protection, DMF (2.0 mD) and triethylamine (0.223 mg, 2.25 mmol) were added. The reaction was stopped at 95°C for 22 hours, and water and EA were added to the system. The system was washed with saturated brine, dried with anhydrous sodium sulfate, and concentrated to dryness under reduced pressure. Column chromatography purification (PE:EA=100:1 to PE:EA=10:1), 330 mg of compound F1035-S2, a yellow solid, yield 71.6%. LCMS 30 (254 nm) #1 FE 95.0%, Rt = 1.968 min; MS calculated value: 307.1; MS measured value: 308.6 [M+H]°. Step 2: F1035-S2 (300 mg, 0.98 mmol WFLA GH, AAR, WAT Slax formic acid (507 mg, 2.94 mmolD)) was stirred overnight at room temperature. The reaction was stopped after LCMS monitoring. The system was concentrated to dryness under reduced pressure and purified by column chromatography (PE:EA=20:1 to PE:EA=2:1), #421 260 mg Compound F1035-S3, a yellow precipitate, had a yield of 78.3%. LCMS (254 nm) showed a purity of 96.0%, Rt = 1.532 min; calculated MS value: 339.1; measured MS value: 35338.2 [MH]. Step 3: Compound F1035-S3 (146 mg, 0.43 mmol JAF 2% trichloroacetic acid in dioxane solution) was added to a solution and heated for 2 hours. The reaction was monitored by LCMS until complete. The solution was concentrated to dryness under reduced pressure to obtain F1035-S4. The crude product was directly used in 99...WO 2024 / 255740 PCT / CN2024 / 098491 Next step. Example 4S: Synthesis of drug linker conjugate STI-F1043 ° FmocH,, from, rmoct, le Hawk person WMS》 fa) HO © Hot person fo) ety no 5 HWE 1: F1043-01 (1.0 g, 2.13 mmol) was dissolved in 4M HCVEA (10.0 mL) and reacted at room temperature for 1.5 hours. After the reaction was monitored by TLC, the solution was concentrated at 45"C to obtain the de-Boc crude product. F1043-02 (220 mg, crude product) and the above de-Boc product (178 mg, 0.48 mL) were added to THEF / H2O (2 mL: 2 mL), JA NaHCO3 (203 mg, 2.42 mmol), VTE for 2 hours. After the reaction was monitored by LCMS, 1M HCl pH was added. Adjust to 3-4, take ethyl acetate (10 mL * 2), dry with saturated brine (10 mL * 2), dry with anhydrous sodium sulfate, concentrate at 45°C, and then directly prepare compound F1043-03 under medium-low pressure reverse phase, which is a transparent oily substance with a yield of 74.9%. LCMS (254 nm, purity 93.6%, Rt = 1.96 min; MS calculated value: 762.3; MS measured value: 763.5 [M+H]*. Step 2: 4 F1043-03 (100 mg, 0.13 mmol) was dissolved in dioxane (20.0 mL) under nitrogen protection, and then NHS (18 mg, 0.15 mmol) and DCC (32 mg, 0.15 mmol) were added. After 15 hours, the reaction was monitored by TLC. The reaction solution was filtered, and the filter cake was washed with DCM (5 mL*3). The filtrate was concentrated at room temperature. The crude product was dissolved in DMF (4 mL) with 1033-02 (90 mg, 0.13 mmol), and DIEA (33 mg, 0.26 mmol) was added. FHEPE for 2 hours, LCMS monitoring of reaction completion.Afterwards, the mixture was concentrated using a 4°C oil pump. The crude product was purified by column chromatography (dioxane / alcohol = 25 / 3) to obtain 110 mg of compound F1043-04 as a yellow solid, with a yield of 58.9%. LCMS (254 nm) showed a purity of 91.2%, Rt = 1.85 min; MS calculated value: 1436.7; MS measured value: 1437.9 [M+H]. Step 3: Compound F1043-04 (110 mg, 0.07 mmol) was added to DMF (2.0 mD), and the mixture was stirred under nitrogen protection for 0.5 hours. The reaction was monitored by LCMS to ensure complete reaction. The reaction solution was concentrated to dryness using a 4°C oil pump, then petroleum ether was added and slurry was added. After filtration, the filter cake and EMCS (16 mg, 0.07 mmol) were added to DMF (2 mL). JA DIEA (19 mg, 0.15 mmOD, nitrogen protection, stirred at room temperature for 2 hours, LCMS monitoring showed complete reaction, concentrated to dryness by oil pump at 45°C, crude product was prepared by high pressure reverse phase (mobile phase: acetylene / 0.05% trifluoroacetic acid aqueous solution), lyophilized to give 7.18 mg of compound STI-F1043, 25" as a yellow solid, yield 6.6%. LCMS (254 nm, purity 99.0%, Rt = 1.71 min; MS calculated value: 1407.7; MS measured value: 1408.6 [M+H]*. 'H NMR (400 MHz, DMSO). 88.81 (s, 1H), 8.64 (t, J = 6.4 Hz, 1H), 8.06 (d, J = 6.8 Hz, 1H), 7.95-7.88 (m, 2H), 7.80-7.72 (m, 2H), 7.64 (d, J = 8.8 Hz, 1H), 7.26 (s, 1H), 6.99 (s, 2H), 5.43 (d, J = 9.6 Hz, 4H), 4.64-4.55 (m, 2H), 4.29-4.15 (m, 3H), 3.58-3.55 (m, 22H), 100 WO 2024 / 255740 PCT / CN2024 / 098491 3.49-3.43 (m, LOH), 3.41-3.33 (m, 5H),3.23 (s, 4H), 3.00-2.98 (m, 2H), 2.47 (s, 4H), 2.28 (J = 6.4 Hz, 2H), 1.89-1.80 (m, 4H), 1.65-1.13 (m, 20H), 0.89-0.81 (m, 9H). The intermediate compound F1043-02 was prepared as follows: [F1043-02a F1043-02 5 F1043-02a (200 mg, 0.48 mmol) / % DCC (120 mg, 0.58 mmol) 34 F — SAF (20.0 mL) F, BRE, FEUILA NHS (67 mg, 0.58 mmol) / % DCC (120 mg, 0.58 mmol), After the reaction was monitored by TLC, the reaction was reversed. Example 46: Preparation of antibody 10 Farletuzumab is an anti-folate receptor of (FoR, FRo) antibody, and its sequence information is as follows: Farletuzumab heavy chain sequence: EVQLVESGGGVVQPGRSLRLSCSASGFTFSGYGLSWVRQAPGKGLEWVAMISSGGSYTYY ADSVKGRFAISRDNAKNTLFLQMDSLRPEDTGVYFCARHGDDPAWFAY WGQGTPVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG 18 LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPOVYTLPPSRDELTKN QVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK 20 Fatazolidone light chain sequence:DIQLTQSPSSLSASVGDRVTITCSVSSSISSNNLH WY QQKPGKAPKPWIYGTSNLASGVPSRF SGSGSGTDYTFTISSLQPEDIATY YCQQWSSYPYMYTFGQGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 25 Trastuzumab is an anti-Her2 antibody, and its sequence information is as follows: Trastuzumab light chain sequence; DIQMTOQSPSSLSASVGDRVTITCRASQDVNTAVAWY QQKPGKAPKLLIYSASFLYSGVPSR FSGSRSGTDFTLTISSLQPEDFATY YCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQL 30 KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSOESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC Trastuzumab heavy chain sequence; EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRY ADSVKGRFTISADTSKNTAYLOMNSLRAEDTAVY YCSRWGGDGFY AMDYWGQGTLVTV 35 SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPELLGGPS VFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFN WY VDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGOPREPOVYTLPPSREEMT KNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ 101 WO 2024 / 255740PCT / CN2024 / 098491 GNVFSCSV MHEALHNHYT QKSLSLSPGK Isotype is an IgG1 antibody against Anti-HEL Human, and its sequence information is as follows: Light chain sequence of Anti-HEL Human IgG1 monoclonal antibody; 5 DIQMTQSPASLSASVGETVTITCRASGNIHNYLA WY QOKOGKSPQLLVYNAKTLADGVPS RFSGSGSGTQYSLKINSLQPEDFGSY YCQHFWSTPRTFGGGTKLEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAK VQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Anti-HEL Human IF) Heavy chain sequence of IgG1 monoclonal antibody: 10 QVQLQESGPGLVRPSQTLSLTCTVSGSTFSGYGVNWVROPPGRGLEWIGMIWGDGNT DYNSALKSRVTMLVDTSKNQFSLRLSSVTAADTAVYYCARERDYRLDYWGQGSLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLOSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPELLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY VDGVEVHNAKTKPREEQYNSTY 15 RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKN QVSLTCLVKGFYPSDIA After constructing the above sequence vector, sequencing confirmed that the constructed sequence was consistent with the above sequence. Expression was performed in Expi293F cells (Thermofisher catalog number; A1452) and Protein A (Cytiva, #5:Purification was performed using 17549801). Example 47: Preparation and detection of antibody-drug conjugates. Antibody-drug conjugates (ADCs) were synthesized using the camptothecin derivative of this invention, and their DAR values ​​were detected. Specifically, purified antibody (SEC-HPLC purity greater than 95%) was added to 10 mM PBS buffer, and 25 μL of 500 mM EDTA was added to bring the EDTA concentration in the reaction system to 2 mM, resulting in an antibody concentration of approximately 1-3 mg / ml. 5-8 equivalents of TCEP (TCEP / mAb, ft PEISS) were added, and the reaction was carried out at 37°C and 600 rpm for 2 hours. After the reaction was complete, the reaction solution was cooled to 49°C, and drug linker conjugate solution (10-16 eq, drug / mAb) was added to DMSO. The mixture was thoroughly mixed, and the reaction was carried out at 44°C and 600 rpm for 1 hour. After the reaction was completed, the reaction solution was added to 30K HEE, YR and reduced to a suitable volume. Then, it was added to a pre-equilibrated desalting column and centrifuged to obtain the purified coupled sample. The concentration, purity, and DAR value of the sample were determined. The DAR value can be detected and calculated using the following method: 1. RP-HPLC detection method: Add 75 μL of 8M hydrochloric acid and 1M Tris to a centrifuge tube, add an appropriate amount of sample (40 μg, converted to volume according to concentration), and finally add water to a final volume of 100 μL. 2 μL of PNA, 1M DTT, *A AOE, F 30°C CA 30 min. After incubation, return to room temperature, 13000 rpm AS min, and transfer the supernatant to a liquid chromatography vial. Use 35" Vanquish high-performance liquid chromatography, with a BioResolve™ RP mAb, Polyphenyl, 450A, 2.7 μm, 2.1*100 mm column (Part No: 186008945, Serial No: 01293033515811), fifi 80°C; DAD detector wavelength 280nm; Wee:Injection rate: 1 ml / min; Injection volume: 20 μg. The positions of the heavy and light chains were then distinguished by comparing the naked antibody with the sample. The sample chromatogram was then integrated to calculate the DAR value. Mobile phase configuration: 102 WO 2024 / 255740 PCT / CN2024 / 098491 Reversed-phase column mobile phase A: 0.05% TFA + 0.05% FA in H2O Reversed-phase column mobile phase B: 0.05% TFA + 0.05% FA in ACN Data analysis: Based on the positions of the light and heavy chains, the chromatogram of the detected sample was integrated to calculate the DAR value. The calculation formula is as follows: 5 Name "Number of Linked Drugs" LC 0 LC+1 2 HC 0 HC+1 2 10 HC+2 4 HC+3 6 LC Peak Area Sum = LC Peak Area T + LC+1 Peak Area HC Peak Area Sum = HC Peak Area + HC+1 Peak Area + HC+2 Peak Area + HC+3 Peak Area LC DAR = X (Number of Linked Drugs * Peak Area Percentage) X LC Peak Area Sum 15 HC DAR = © (Number of Linked Drugs * Peak Area Percentage) X HC Peak Area Sum DAR = LC DAR + HC DAR 2. SEC-HPLC Detection Method: Dilute the sample to 1 mg / mL with ultrapure water, injection volume 20 μL, use Vanquish high performance liquid chromatography, chromatographic column selected XBridge@ BEH200A SEC 3.5um, 7.8*150 mm Column (Part No. 186007639, Serial No. 20 01783201716107); Column temperature 25°C; DAD detector wavelength 280nm; Flow rate: 0.5 min. Approximately 25 μg of protein was diluted to 0.5 μg / bL with 50 mM NH4OHCO3, and 0.5 μL of PNGaseF and 1 RL 1 M DDT were added. 0.5 ng of calcium was injected for mass spectrometry analysis. A Waters Xevo G2-XS Q-TOF column was used, with an ACQUITY UPLC BEH C4 column, 1.7 pam, 2.1 x 50 mm.mm, 300 Å (186004495), HEIL 80°C; DAD 25 " detector wavelength 280 nm; flow rate: 0.3 ml / min; DAR fi was calculated based on the response signal of the sample mass spectrum. The antibody-drug conjugate of the present invention was synthesized by the above method and its DAR value was measured, and the results are shown in Table 1. Table 1. Prepared ADCs and their characterization ADC name | Contained drug moiety | Average DAR value of ADC structure Fa-ADC1 | STI-G 7.0 Fa-ADC2 | STI-A3 7.2 WO 2024 / 255740 PCT / CN2024 / 098491 Fa-ADC3 | STI-F3 7.1 Fa-ADC4 | STI-F4 7.1 Fa-ADC5 | STI-FL002 5.8 Fa-ADC6 | STI-F1001 7.2 Tra-ADC7 | STI-F1006 7.9 Tra-ADC8 | STI-F1010 Tra-ADC9 | STI-F1011 Tra-ADC10 | STI-F1012 7.5~~ 6 H 0 2 4H N H3C F n 104 WO 2024 / 255740 PCT / CN2024 / 098491 Tra-ADC11 | STI-F1013 9 nt \ H N=N H N “~o NH。 HN i Tra-ADC12 | STI-F1014 7.6 fe} H i i { N ony nv \} ° mAb CoAy y a a Tra-ADC13 | STI-F1015 fe , 。 7.7 六 一TY CC ee D es HO Tra-ADC14 | STI-F1018 mAb~>_N Q 2 Ne! ° ° ° ° H NM Vee re Tra-ADC15 | STI-F1019 an 9 。 a Nhe wy stant ert mAb N Hse Tra-ADC16 | STI-G1001 78 mAb-)>N MeN i y HL HA, k SNe oy y oY N oy vp, Tra-ADC17 | STI-F1031 74 Q io) O , £0 Oy NM VZ = mAb Cee EeCe OC Ceres: a | Ho ° ° ool; oF OH N HaC Tra-ADC18 | STI-F1034 75 9 do O 吕 mAb Coe eeeeeeeus NAN ; ‘Oo 8 OpAr Ao, Too 5 8 “TH aa Ho N HaC Tra-ADC19 | STI-F1035 78 mAb (Ns pe 5 Q { O © ooh To =H N 105 WO 2024 / 255740 PCT / CN2024 / 098491 Tra-ADC20 | STI-F1043 75 ; Kl pes ; {0 N,,, N =~ N Lo NH 3 ay H3C if Iso-ADC21 | STI-F1006 Q 0 78 y nv ~~ mAb yy nai Ay eal Iso-ADC22 | STI-F1011 75 9 H H 人 vn \ 2 ° mAb ae A a —= 46 . N Iso-ADC23 | STI-F1012 73 mAb. AN Q O WI H 9 4 vn \ O S N NA UN a = HO H 0 2 4H HC F n Iso-ADC24 |STI-F1014 5 H ° H i N hoa, nN \ ; 5 mAb yO oa ren aan Ope Iso-ADC25 | STI-F1031 78 Q ° ° H x? 2 o fe) nN \-+¥ ° mAb fe AOZN Sa ° ° oA Ag No FR N Fa-ADC @ | (MC-GGFC- o 1 9 H 24 9 7.0 tk DXD) mAb Wy OS yoy 囗 re) \ .NH 5 . acon F Nw \ yy wy HO"; ot n Tra-ADC & | (MC-GGFC- o a) 4H on 9 tk DXD) mAb Wy OS yoy oO oO \ ANH b . O MG F Nw \ yy wy HO"; ot n 106 WO 2024 / 255740 PCT / CN2024 / 098491 Iso-ADC & | (MC-GGFC- 0 4 4 2 y 0 7.7 ratio | DXD) mab yy BAe O Cr ; 5. 1) In Fa-ADCs, mAb represents futuximab (Fa); in Tra-ADCs, mAb represents trastuzumab (Tra); in Iso-ADCs, mAb represents IgG1 antibody of Anti-HEL Human, and mn represents an integer from 0 to 8, such as 0, 1, 2, 3, 4, 5, 6, 7 or 8. 5 2) The drug-linker conjugate MC-GGFC-DXD in ADC reference is purchased from Shanghai Gangyuan Biomedical Technology Co., Ltd. Biological Section Examples A. Camptothecin derivative toxin killing assay Tumor cells growing in logarithmic phase (human ovarian cancer cell line SK-OV-3; human colon cancer cell line HCT116; human lung adenoma cell line A549) are digested, and the cell density is adjusted to about 3×104 cells / mL. Plate the cells at about 100 µL per well into a white 96-well plate. The camptothecin derivative toxins of the examples of the present application are subjected to gradient dilution with corresponding cell culture medium, then mixed with the cells, and cultured in an incubator at 37°C and 5% CO2 for 3 days.The growth inhibition of camptothecin derivative toxins on tumor cells was assessed using the Cell Titer Turbo 2.0 Luminescent Cell Viability Assay (Damas life, RA-GLI1T). Relative light unit (RLU) values ​​were read from cell culture plates using a microplate reader. The ICso was calculated using Graphpad Prism 8.0 software to analyze the experimental data. The above experiments were repeated using the following compounds as positive controls: 0 Fo — N asNH HO n7 \ / Oo ' N oO HO 2 0 / " AU¥ oO SN-38 DXD. The results are shown in Table 2. It can be seen that the compounds of the embodiments of this application exhibit good tumor cell killing activity. In particular, the tumor cell killing activity of the compounds of Examples 3 (STI-A), 4 (STI-A2), 6 (STI-E-05), 8 (STI-F-8), 10 (STI-F-03C-1), 13 (STI-G2-06C), and 16 (CSTI-F) is comparable to that of SN-38 and Dxd, and the tumor cell killing activity of the compounds of Examples 11 (STI-GJD), 14 (STI-G2), 17 (STI-F6), and 19 (STILF13) is even significantly better than that of SN-38 and Dxd. Table 2 The killing activity of camptothecin derivatives against tumor cells 7 hs Aa oes 2 A a mre 107 WO 2024 / 255740 PCT / CN2024 / 098491 esterase asf asters [eases t raia fae NT indicates not detected. Example A: Camptothecin derivative toxin killing experiment The camptothecin derivatives of the example were subjected to gradient dilution with DMSO and the corresponding cell culture medium and then added to a white plate with 96 FL 5, and tumor cells (human ovarian tumor cell line SK-OV-3; A Sir AHH & HCT116; human lung adenocarcinoma cell line AS49; human lung) in the logarithmic growth phase were tested.Adenoma cell line NCI-H441 (human oral epidermoid carcinoma KB) was digested and the cell density was adjusted to approximately 3.33 x 10⁷ mL. Cells were seeded at 90 hL / well into 96-well plates with pre-added samples and cultured at 37°C in a 5% CO₂ incubator for 3 days. The inhibitory effect of small molecule compounds on tumor cell growth was evaluated using the Cell Titer Turbo 2.0 Luminescent Cell Viability Assay (Damas life, RA-GLIDT). The relative light unit (RLU) values ​​were read from the cell culture plates using a microplate reader. The IC₀ values ​​were calculated using Graphpad Prism 8.0 software. The results are shown in Table 3. Table 2 shows the cytotoxic activity of camptothecin derivatives against tumor cells. NT indicates no detection. 15” Example B. ADC Killing Assay 108 WO 2024 / 255740 PCT / CN2024 / 098491 After digesting tumor cells in the logarithmic growth phase, the cell density was adjusted to approximately 1-10 x 10⁴ / mL. Cells were seeded at 50 wL / well in 96-well plates with a white background and cultured overnight at 37°C with 5% CO₂. The ADCs or reference ADCs of this application were serially diluted with the corresponding cell line used for cell culture and mixed with the cells, and cultured at 37°C with 5% CO₂ for 5-7 days. The growth inhibition of tumor cells was demonstrated by the Cell Titer Turbo 2.0 Luminescent Cell Viability Assay (Damas life, 5 RA-GLII) iF RE ADC. The relative light unit (RLU) values ​​were read on the cell culture plate using a microplate reader. The experimental data were analyzed using Graphpad Prism 8.0 software. ICso was calculated. The results are shown in Table 3. Fa-ADC1 showed cytotoxic activity against oral epidermal-like KB tumor cells.Significantly superior to the positive control Fa-ADC reference. Table 3. Killing activity of ADC against human oral epidermal-like KB tumor cells. 10 N / A indicates not applicable. In addition, Her2 endogenous expression tumor cell lines NCI-N87 (gastric cancer cell line, Nanjing Kebai, CBP60491) and Capan-1 (pancreatic cancer cell line, Nanjing Kemen, CBP60543) were used for in vitro drug killing experiments. NCI-N87 and Capan-1 cells were obtained by digestion with trypsin (Gibco, 25200072). Resuspend NCI-N87 cells in RPMI-1640 medium containing 15% fetal serum (FEBS, Gibco, 10091-148), adjusting the cell concentration to 2 x 10⁴ cells / mL, adding 100 pL per well (2000 cells) to a white-bottomed clear 96-well microplate (Perkin Elmer, 6005181). Resuspend Capan-1 cells in RPMI-1640 medium containing 20% ​​FBS, adjusting the cell concentration to 4 x 10⁴ cells / mL, adding 100 hbL per well (4000 cells) to a white 96-well microplate. Add 200 μL of DPBS to the periphery of each well of the 96-well microplate. Incubate the 96-well microplate at 37°C in a CO₂ incubator for 1 day. Add 20% FBS... RPMI-1640 de Fe Ses BE ORE AY SCH A) ADCs or reference ADCs. For NCI-N87 cells, the initial concentration was 600 nM (6*), 5-fold serial dilution, DAAEFL 20 wh added to the 96-well plate from the previous step, with a total volume of 120 hL per well. For Capan-1 cells, the initial concentration was 3000 nM (6*), 5-fold serial dilution, 20 pL per well added to the 96-well plate from the previous step, with a total volume of 120 hL per well. The above 96-well plates were placed in a 37°C CO2 incubator and incubated for 4-5 days. On day 25,Remove the 96-well plate containing Capan-1 from the incubator and the Cell Titer Turbo 2.0 (Adamas lift, RA-GLID reagent) from the refrigerator, allowing it to return to room temperature. Then add 60 μL of 96-well FLARE (HD) per well and incubate for 10 minutes. After 10 minutes, read the fluorescence signal on a Microplate Reader (BMG, CLARIOstar Plus). On day 6, remove the 96-well plate containing NCLN87 from the incubator and the Cell Titer Turbo 2.0 reagent from the refrigerator, allowing it to return to room temperature. Then add 60 μL of FLARE per well and incubate for 10 minutes at room temperature. After 10 minutes, read the fluorescence signal on a Microplate Reader. See Figures 2a to 2d (ADC drug killing experiment on Capan-1) and 2a to 2d (ADC drug killing experiment on NCLN87). As shown in the killing experiment, the ADC drug of the present invention exhibited in vitro killing activity in both NCLN87 and Capan-1. Example C, the bystander effect of ADCs will be adjusted to a suitable cell density after digestion of antigen-positive cells (folate receptor wx-positive cells KB) and antigen-negative cells (293T cells) in logarithmic growth phase. Antigen-positive and antigen-negative cells were added to 96-well plates containing 5% FLA ARP, BF2, 37°C, 5% CO2 incubator and cultured overnight. The next day, the ADC was serially diluted with the corresponding cell culture medium and added at 100 hL / well to 96-well plates containing cells, and cultured at 37°C, 5% CO2 incubator for 5-7 days. The results were obtained using Cell Titer Turbo 2.0. Luminescent Cell Viability Assay (Adamas life,The RA-GLII-A)VF fh ADC inhibited the growth of tumor cells. The relative light unit (RLU) values ​​were read on the cell culture plate using a microplate reader. The ADCs of this application exhibit a strong labor observer effect. CHOK1-EGFP cells (a subline of Chinese hamster oocytes, target-negative, laboratory-constructed) and OE19 cells (human esophageal cancer cells, target-positive, BME CBP60495) were obtained by trypsin digestion, resuspended, and counted. OE19 cells were seeded into 96-well plates at a density of 0-8000 cells / well, and CHOK1-EGFP cells were seeded into 96-well plates at a density of 500 cells / well. KK ADCs or reference ADCs were added to 96-well plates at a density of 1 μg / mL. The 96-well plates were placed in an Incucyte assay instrument and incubated for 7 days. The green light signal was monitored in real time. Data were collected after 7 days and plotted to assess the bystander effect of ADCs. The data are ...

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