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KR-20260067388-A - Multidrug linker and antibody-drug conjugate

KR20260067388AKR 20260067388 AKR20260067388 AKR 20260067388AKR-20260067388-A

Abstract

This specification relates to a multidrug linker, an antibody-drug conjugate comprising said multidrug linker, a method for manufacturing an antibody-drug conjugate, and the use thereof for treating diseases. The specified structure of the present invention effectively reduces the aggregation of the multi-payload drug-antibody conjugate, which is advantageous for process scaling. Furthermore, the payload drugs exhibit a significant synergistic effect, thereby obtaining a synergistic antitumor effect, effectively overcoming drug resistance, and improving the therapeutic window of the drugs.

Inventors

  • 쑨 원룽
  • 멍 쉰
  • 스 위

Assignees

  • 상하이 화오 컴퍼니 리미티드

Dates

Publication Date
20260512
Application Date
20240805
Priority Date
20230804

Claims (20)

  1. In a compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, The above compound includes a structure represented by Formula I, but (I) Q is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of a direct bond, an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from a halogen, a hydroxyl group, an amino group, a carboxyl group, a sulfonic acid group, a sulfonate group, a phosphate group, and an alkoxy group; B1 and B2 are independently direct-bonded or branched groups, and the condition is that when C1 is directly bonded, B1 is not directly bonded; P1 , P2 , and P3 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 , T2, and T3 are each independently directly bonded or arbitrarily substituted spacer groups, and the condition is that at most two of T1 , T2 , and T3 are directly bonded and at least one of B2 , T1 , T2 , and T3 is substituted by a hydrophilic group; D1 , D2 , and D3 are the first drug unit, the second drug unit, and the third drug unit, respectively, and are identical or different; a and b are independently 0, 1, 2, or 3, and the condition is that a and b are not simultaneously 0; When a=0, B1 is a direct coupling and T1 is not a direct coupling; When b=0, B1 is a compound that is not directly bonded, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  2. In paragraph 1, The above compound includes a structure represented by formula Ia, but (Ia) Q is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of a direct bond, an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from a halogen, a hydroxyl group, an amino group, a carboxyl group, a sulfonic acid group, a sulfonate group, a phosphate group, and an alkoxy group; B 1 is a branched group; P1 and P2 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 and T2 are each independently directly bonded or arbitrarily substituted spacer groups, and the condition is that at least one of T1 and T2 is directly bonded and at least one of T1 and T2 is substituted by a hydrophilic group; D1 and D2 are the first drug unit and the second drug unit, respectively, and are identical or different; a is a compound, which is 1, 2, or 3, or its tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  3. In paragraph 1, The above compound includes a structure represented by the formula Ib, but (Ib) Q is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from halogen, hydroxyl group, amino group, carboxyl group, sulfonic acid group, sulfonate group, phosphate group, and alkoxy group; B 2 is a branched group; P1 and P3 are each independently directly bonded or arbitrarily substituted polypeptide residues or glucose fragments; T1 is an arbitrarily substituted spacer group, T3 is a directly bonded or arbitrarily substituted spacer group, and at least one of B2 and T3 is substituted by a hydrophilic group; D1 and D3 are the first drug unit and the third drug unit, respectively, and are identical or different; b is a compound that is 1, 2, or 3, or its tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  4. In paragraph 1, B1 and B2 are independently branched groups; P1 , P2 , and P3 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 is an arbitrarily substituted spacer group, T2 and T3 are each independently directly bonded or arbitrarily substituted spacer groups, and at least one of B2 , T2 , and T3 is substituted by a hydrophilic group; D1 , D2 , and D3 are the first drug unit, the second drug unit, and the third drug unit, respectively, and are identical or different; a and b are independently 1, 2 or 3, a compound, or its tautomer, meso compound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  5. In any one of paragraphs 1 through 4, Q is arbitrarily substituted (Q1), arbitrarily substituted (Q2), arbitrarily substituted (Q3), arbitrarily substituted (Q4), arbitrarily substituted (Q5), arbitrarily substituted (Q6), arbitrarily substituted (Q7), arbitrarily substituted (Q8), arbitrarily substituted A compound , or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the group consisting of (Q9), where * indicates a position connected to C1, and in which the W group of Q7 is bromine, Ar-S group, Ar is a phenyl group or a substituted phenyl group, and the substituent of the substituted phenyl group is selected from alkyl groups, alkoxy groups, halogens, ester groups, nitro groups, and -C(O)NR1R2-, and R1 and R2 are independently selected from chemical bonds, H and alkyl groups, or R1 and R2 form a 5-7-membered heterocycle, and the heteroatom is selected from one or more of O, N and S.
  6. In any one of paragraphs 1 through 5, C1 is directly bonded, -( CH2 ) m1- , -( CH2 ) m1- (O- CH2 - CH2 ) m2- , -( CH2 ) m1- (O- CH2 -CH2) m2- (CH2) m3- , -( CH2 ) m1 -C(O)NH-( CH2 ) m2- , -( CH2 ) m1 - NHC(O)-( CH2 ) m2- and -(C A compound selected from C-(CH 2 ) m1 - wherein m1, m2 and m3 are independently integers from 1 to 6, and the left side of the indicated substituent is connected to Q and the right side is connected to B 1 or P 1 , or a tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound form thereof, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.
  7. In any one of paragraphs 1 through 6, B 1 is directly coupled, and Selected from a group consisting of; Preferably and and, junction point 1 is connected to C1 or Q (when C1 is directly bonded), junction point 2 is connected to P1 , junction points *, 3, 4 and 5 are connected to P2 , L1 is selected from -( CH2 ) m1- , -( CH2 ) m1 -C(O)NH-( CH2 ) m2- , and -( CH2 ) m1 -NHC(O)-( CH2 ) m2- , L2 , L3 and L4 are independently -( CH2 ) m1- (O- CH2 - CH2 ) m2- or -( CH2 ) m1- (O- CH2 - CH2 ) m2- ( CH2 ) m3- , L5 is -( CH2 ) m1- , and L6 is -( CH2 ) m1- or -(CH 2 -CH 2 -O)m 2 and m1, m2, m3, w and v are each independently 1, 2, 3, 4, 5 or 6; Preferably, when C1 is a direct bond, B1 is or Phosphorus, a compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  8. In any one of paragraphs 1 through 7, B 2 is , and A compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer , or deuterium compound form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the group consisting of the following, wherein linkage point 1 is connected to T1 , linkage point 2 is connected to P3, linkage point 3 is connected to the hydrophilic group, and M is selected from the group consisting of an optionally substituted C1 - C6 alkylene group, an optionally substituted C1-C6 alkoxy group, or an optionally substituted C1-C5 alkenylene group.
  9. In any one of paragraphs 1 through 8, T1 , T2 , and T3 are independently and directly coupled, and arbitrarily substituted A compound selected from the group consisting of, wherein linkage 1 is connected to P1 , P2 or P3 and linkage 2 is connected to D1 , D2 or D3 , or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound form thereof, or a mixture form thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.
  10. In any one of paragraphs 1 through 9, P1 , P2 , and P3 comprise a polypeptide residue composed of an optionally substituted amino acid selected from phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine; preferably, P1 , P2 , and P3 comprise a polypeptide residue composed of an optionally substituted amino acid selected from glycine, phenylalanine, valine, alanine, arginine, citrulline, aspartic acid, asparagine, and lysine; More preferably, P1 , P2 , P3 are optionally substituted phenylalanine-lysine (Phe-Lys), valine-alanine (Val-Ala), valine-citrulline (Val-Cit), glutamic acid-valine-alanine (Glu-Val-Ala), glutamic acid-valine-citrulline (Glu-Val-Cit), valine-lysine (Val-Lys), alanine-alanine (Ala-Ala), alanine-alanine-alanine (Ala-Ala-Ala), alanine-alanine-asparagine (Ala-Ala-Asn), alanine-leucine (Ala-Leu), leucine-leucine (Leu-Leu), phenylalanine-arginine (Phe-Arg), phenylalanine-lysine (Phe-Lys), (cBu-Cit), , , comprising polypeptide residues selected from glycine-phenylalanine-glycine (Gly-Gly-Phe-Gly) and glycine-proline (Gly-Pro); Or, one of the P 2 - T 2 combination or the P 3 - T 3 combination A compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein linkage point 1 is connected to B1 or B2 and linkage point 2 is connected to D2 or D3.
  11. In any one of paragraphs 1 through 10, The hydrophilic group is selected from substituted polysarcosine residues, polyglycerols, polyols, glycosyl groups, cyclodextrins, substituted ethylene glycol fragments, substituted glycosylated polyethylene glycol, substituted glycosylated polyglycerols, substituted cyclodextrin polyethylene glycol, or combinations thereof; Preferably, the substituted polysarcosine residue is and, n 2 is an integer between 4 and 20, for example, an integer between 4 and 16, and R is selected from C 1 -C 6 alkyl groups, C 1 -C 6 cycloalkyl groups, C 1 -C 6 alkoxy groups; Preferably, the substituted glycosylated polyethylene glycol fragment is and, n 3 is an integer between 4 and 18, for example, an integer between 4 and 12; Preferably, the substituted glycosylated polyglycerol fragment is and, n 4 is an integer between 4 and 12, for example, an integer between 4 and 10; Preferably, the substituted polyethylene glycol fragment is and, n 5 is an integer between 4 and 24, for example, an integer between 8 and 18; Preferably, at least one of T1 , T2 , and T3 and X is arbitrarily substituted A compound, or its tautomer, mesocompound, racemic mixture, enantiomer , diastereomer , or deuterium compound form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, wherein the hydrophilic group is connected to T1 , T2 , or T3 through X, connection point 1 is connected to the hydrophilic group, and connection point 2 is connected to one of T1, T2, or T3.
  12. In any one of paragraphs 1 through 11, D1 , D2 , and D3 are a first anticancer agent, a second anticancer agent, and a third anticancer agent; preferably, at least two of the first anticancer agent, the second anticancer agent, and the third anticancer agent have a synergistic anticancer effect; Preferably, D1 , D2 , and D3 are independently cytotoxic drugs or tumor-targeted therapeutic drugs; preferably, at least one of D1 , D2 , and D3 is a cytotoxic drug, and at least one other is a tumor-targeted therapeutic drug; more preferably, at least two of D1 , D2 , and D3 are tumor-targeted therapeutic drugs, for example, tumor-targeted therapeutic drugs capable of causing synergistic or overlapping effects; Preferably, the cytotoxic drug is selected from a drug unit targeting topoisomerase, a drug unit targeting tubulin, a nucleoside antimetabolite anticancer drug unit, and a drug unit targeting DNA; Preferably, the tumor-targeted therapeutic drug is selected from drug units targeting DNA damage response (DDR) or “synthetic lethality” related pathways, drug units targeting epigenetics, drug units targeting apoptosis-related pathways, drug units targeting transcription factors, and drug units targeting immune activation pathways, and comprises EFGR pathway-related target inhibitors; Ras-Raf-MAPK pathway inhibitors; PI3K/AKT/mTOR pathway inhibitors; cell cycle pathway inhibitors; cGAS-STING signaling pathway agonists; estrogen receptor antagonists; androgen receptor antagonists; glucocorticoid receptor modulators; autophagy inhibitors; FAK inhibitors; Smo inhibitors; BTK inhibitors; PDE4 inhibitors; Lck inhibitors; PLK1 inhibitors; TLR7/8 modulators; and N-myristiltransferase (NMT) inhibitors; Preferably, the drug unit targeting the topoisomerase is a topoisomerase I (TOPO1) inhibitor or a topoisomerase II (TOPO2) inhibitor, and Exatecan (CAS 171335-80-1), DXd (CAS 1599440-33-1), 7-ethyl-10-hydroxycamptothecin (SN38, CAS 86639-52-3), Belotecan (CAS 213819-48-8), (4- NH2 )-Exatecan (AZD'0132, CAS 2495742-21-5), 7-MAD-MDCPT (CAS 765871-81-6), 7-aminomethyl-10-methyl-11-fluorocamptothecin (CAS 2378616-23-8), vorelloxin (CAS 175414-77-4) or derivatives and analogs thereof; Preferably, the drug unit targeting tubulin is Eribulin, vinblastine, paclitaxel, MMAE, MMAF, mytansine, or a derivative thereof; Preferably, the nucleoside antimetabolite anticancer drug is gemcitabine or decitabine; Preferably, the DNA-targeting drug unit is a DNA subcompartment conjugate (PBD), DNA alkylating agent (Duocarmycin), Trabectedin, Rubitidine, or a derivative thereof; Preferably, the drug unit targeting the DNA damage response (DDR) or “synthetic lethality” related pathway is a PARP inhibitor, an ATR inhibitor, a CHK1 inhibitor, an ATM inhibitor, a DNA-PK inhibitor, a WEE1 inhibitor, a POLQ inhibitor, a CDK12 inhibitor, a USP1 inhibitor, a PKMYT1 inhibitor, or a Rad51 inhibitor; The above PARP inhibitor is preferably Rucaparib, Niraparib, Veliparib, A-966492, Talazoparib, AZD5305, Venadaparib, Mefuparib, or an analog thereof; the above ATR inhibitor is preferably Berzosertib, Ceralasertib, or an analog thereof; and the above CHK1 inhibitor is preferably Prexasertib (LY2606368), AZD7762, Rabusertib (LY2603618), MK-8776 (SCH 900776), CHIR-124, PF-477736, CCT245737 (SRA737, PNT-737), GDC-0575 (ARRY-575), or an analog thereof; and/or the WEE1 inhibitor is preferably ZN-C3 (CAS 2376146-48-2), Adavosertib (AZD1775, CAS 955365-80-7), Debio 0123 (CAS 2243882-74-6) or an analogue thereof; Preferably, the analogue of the AZD5305 is or It contains, wherein R a is selected from -C(O)NH-R b or -NHC(O)-R b , and R b is selected from a C 2-7 alkyl group, a C 3-7 monocyclic alkyl group, a C 4-10 dicyclic alkyl group, a C 5-9 spirocyclic cycloalkyl group, and a C 5-9 cross-linked cycloalkyl group substituted by at least one primary amine group or secondary amino group, or R b is a 4-6-membered monocyclic heterocycloalkyl group containing 1-5 nitrogen, oxygen, and sulfur atoms, a C 4-10 dicyclic heterocycloalkyl group, a C 5-9 spirocyclic heterocycloalkyl group, or a C 5-9 cross-linked heterocycloalkyl group, and the R b group contains at least one primary amine group or secondary amine group; Preferably, the AZD5305 analog is selected from the following structures, and Preferably, the PKMYT1 inhibitor is preferably RP-6306 and GSK-1520489A or analogs thereof; Preferably, analogs of the WEE1 inhibitors Adavosertib (AZD1775, CAS 955365-80-7) and Debio 0123 (CAS 2243882-74-6) are selected from the following structures, and Preferably, the drug unit targeting the apoptosis-related pathway is a Bcl-2 family protein inhibitor; said Bcl-2 family protein inhibitor includes BCL-2 inhibitors, BCL-XL inhibitors, and MCL-1 inhibitors, and preferably is Venetoclax (CAS 1257044-40-8), Navitoclax (CAS 923564-51-6), Navitoclax analog (CAS 2143096-93-7), ABT-737 (CAS 852808-04-9), A-1331852 (CAS 1430844-80-6), S64315 (CAS 1799631-75-6) or an analog thereof; Preferably, the A-1331852, S64315 analog is selected from the following structures, and Preferably, the drug unit targeting the epigenetics is an LSD1 inhibitor, an EZH2 inhibitor, a BRD4 inhibitor, a PRMT5 inhibitor, or a PRMT1 inhibitor; The above LSD1 inhibitor is preferably Tranylcypromine, ORY-1001 (Iadademstat, CAS 1431303-72-8), CC-90011 (Pulrodemstat, CAS 1821307-10-1), ORY-2001, GSK-2879552, IMG-7289, INCB059872, or TAK-418 or an analog thereof; the above EZH2 inhibitor is preferably Tazemetostat (CAS 1403254-99-8), GSK2816126, CPI-1205, PF-06821497, SHR2554, XNW5004, HH2853 or an analog thereof; and the above BRD4 inhibitor is a BI-2536 analog, Birabresib; The PRMT5 inhibitor is preferably GSK3326595, AMG 193, MRTX1719, SKL27969, TNG908, SCR-6920, SH3765, SYHX2001, or an analog thereof; the PRMT1 inhibitor is preferably GSK3368715 (CAS 1629013-22-4), MS023 (CAS 1831110-54-3), or an analog thereof; Preferably, the drug unit targeting the immune activation pathway is a PD-L1 inhibitor, a CBL-B inhibitor, a TLR7/8 agonist, a PTPN2/1 inhibitor, or a STING agonist; Preferably, the inhibitor of the Ras-Raf-MAPK pathway is an SOS1 inhibitor BAY-293 (CAS 2244904-70-7), BI-3406 (CAS 2230836-55-0) or an analog thereof; a KRAS inhibitor BI-2493 (CAS 2937344-16-4), MRTX1133 (CAS 2621928-55-8) or an analog thereof; MEK inhibitors Cobimetinib (CAS 934660-93-2), Selumetinib (CAS 606143-52-6), Mirdametinib (CAS 391210-10-9), Binimetinib (CAS 606143-89-9), TAK-733 (CAS 1035555-63-5), GDC-0623 (CAS 1168091-68-6), AZD8330 (CAS 869357-68-6), Trametinib (CAS 871700-17-3), Trametiglue (CAS 2666940-97-0) or analogs thereof; comprising ERK inhibitor ASN007 (CAS 2055597-12-9) or analogs thereof; Preferably, the cell cycle pathway inhibitor comprises the CDK4/6 inhibitors Palbociclib (CAS 571190-30-2), Abemaciclib (CAS 1231929-97-7), Ribociclib (CAS 1211441-98-3), Dalpiciclib (CAS 1637781-04-4) or analogs thereof; the pan-CDK inhibitor Dinaciclib (CAS 779353-01-4) or analogs thereof; Preferably, the PI3K/AKT/mTOR pathway inhibitor comprises PKI-587 (CAS 1197160-78-3), demethylated PKI-587 (CAS 1950569-63-7), AZD8055 (CAS 1009298-09-2), Afuresertib (CAS 1047644-62-1), or an analogue thereof; Preferably, the estrogen receptor antagonist/degrader Fulvestrant (CAS 129453-61-8), Elacestrant (CAS 1349723-93-8); androgen receptor antagonist abiraterone (CAS 154229-19-3), JNJ-63576253 (CAS 2110428-64-1) or an analogue thereof; Preferably, the autophagy inhibitor is hydroxychloroquine (HCQ, CAS 118-42-3) and chloroquine (CQ, CAS 54-05-7) or an analogue thereof; Preferably, the drug targeting the transcription factor is a GSPT1 degrader (molecular gel), and the GSPT1 degrader is selected from the following structures, and Preferably, the FAK inhibitor is Ifebemtinib (CAS 1227948-82-4), Defactinib (CAS 1073154-85-4), PF-562271 (CAS 717907-75-0) or an analog thereof, and is selected from the following structures, Preferably, the PLK1 inhibitor is BI2536 (CAS 755038-02-9), Volasertib (CAS 755038-65-4), GSK461364 (CAS 929095-18-1), Onvansertib (CAS 1034616-18-6) or an analogue thereof; Preferably, the N-myristiltransferase (NMT) inhibitor is MYX1715 (CAS 2445448-66-6) or an analog thereof; More preferably, D1 , D2 , and D3 are selected from combinations of the following table, and More preferably, (i) at least one of D1 , D2 and D3 is a cytotoxic drug selected from drugs targeting topoisomerase, e.g., a TOPO1 inhibitor, and at least another is a tumor-targeted therapeutic drug, wherein the tumor-targeted therapeutic drug is a drug targeting DNA damage response (DDR) or “synthetic lethality” related pathways, and is selected from PARP inhibitors, ATR inhibitors, CHK1 inhibitors, ATM inhibitors, DNA-PK inhibitors, WEE1 inhibitors, POLQ inhibitors, CDK12 inhibitors, USP1 inhibitors, PKMYT1 inhibitors, and Rad51 inhibitors; (ii) At least two of D1 , D2 and D3 are cytotoxic drugs and are independently selected from drug units targeting topoisomerase and drug units targeting tubulin, e.g., TOPO1 inhibitors, TOPO2 inhibitors and tubulin inhibitors; (iii) At least two or all three of D1 , D2 and D3 are tumor targeted therapeutic drugs and are independently selected from drug units targeting DNA damage response (DDR) or “synthetic lethality” related pathways, drug units targeting epigenetics, drug units targeting apoptosis-related pathways, drug units targeting cell cycle pathways, drug units targeting transcription factors, and drug units targeting immune activation pathways, wherein the targets include EFGR pathway-related target inhibitors; Ras-Raf-MAPK pathway inhibitors; PI3K/AKT/mTOR pathway inhibitors; cell cycle pathway inhibitors; cGAS-STING signaling pathway agonists; estrogen receptor antagonists; androgen receptor antagonists; glucocorticoid receptor modulators; autophagy inhibitors; FAK inhibitors; Smo inhibitors; BTK inhibitors; PDE4 inhibitors; Lck inhibitors; PLK1 inhibitors; TLR7/8 modulators; and N-myristiltransferase (NMT) inhibitors; (iv) One of D1 , D2 and D3 is a cytotoxic drug, said cytotoxic drug is selected from drug units targeting topoisomerase and drug units targeting tubulin, e.g., TOPO1 inhibitors or tubulin inhibitors, and the other one or two are tumor targeted therapeutic drugs, said tumor targeted therapeutic drugs are selected from drug units targeting DNA damage response (DDR) or “synthetic lethality” related pathways, drug units targeting epigenetics, drug units targeting apoptosis-related pathways, drug units targeting cell cycle pathways, drug units targeting transcription factors, and drug units targeting immune activation pathways, and the targets are EFGR pathway-related target inhibitors; Ras-Raf-MAPK pathway inhibitors; PI3K/AKT/mTOR pathway inhibitors; cell cycle pathway inhibitors; cGAS-STING signaling pathway agonists; estrogen receptor antagonists; androgen receptor antagonists; glucocorticoid receptor modulators; autophagy inhibitors; FAK inhibitors; Includes SMO inhibitors; BTK inhibitors; PDE4 inhibitors; Lck inhibitors; PLK1 inhibitors; TLR7/8 modulators; and N-myristiltransferase (NMT) inhibitors; More preferably, D1 , D2 and D3 are compounds, or tautomers, mesocompounds, racemic mixtures, enantiomers, diastereomers, or deuterium forms thereof, or mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof, selected from the combinations shown in Table A.
  13. In any one of paragraphs 1 through 12, The above compound is a compound selected from Formulas A1 to A190 , or its tautomer, meso compound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.
  14. In a compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate, The above compound includes a structure represented by Formula II, but, ( II) Q' is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of a direct bond, an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from a halogen, a hydroxyl group, an amino group, a carboxyl group, a sulfonic acid group, a sulfonate group, a phosphate group, and an alkoxy group; B1 and B2 are independently direct-bonded or branched groups, and the condition is that when C1 is directly bonded, B1 is not directly bonded; P1 , P2 , and P3 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 , T2, and T3 are each independently directly bonded or arbitrarily substituted spacer groups, and the condition is that at most two of T1 , T2 , and T3 are directly bonded and at least one of B2 , T1 , T2 , and T3 is substituted by a hydrophilic group; D1 , D2 , and D3 are the first drug unit, the second drug unit, and the third drug unit, respectively, and are identical or different; a and b are independently 0, 1, 2, or 3, and the condition is that a and b are not simultaneously 0; When a=0, B1 is a direct coupling and T1 is not a direct coupling; When b=0, B1 is a compound that is not directly bonded, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  15. In Paragraph 14, The above compound includes a structure represented by Formula IIa, but, (IIa) Q' is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of a direct bond, an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from a halogen, a hydroxyl group, an amino group, a carboxyl group, a sulfonic acid group, a sulfonate group, a phosphate group, and an alkoxy group; B 1 is a branched group; P1 and P2 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 and T2 are each independently directly bonded or arbitrarily substituted spacer groups, and the condition is that at least one of T1 and T2 is directly bonded and at least one of T1 and T2 is substituted by a hydrophilic group; D1 and D2 are the first drug unit and the second drug unit, respectively, and are identical or different; a is a compound, which is 1, 2, or 3, or its tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  16. In Paragraph 14, (IIb) Q' is a functional group that can be conjugated to cysteine, lysine, non-natural amino acids, or glycosyl groups of an antibody molecule; C1 is selected from the group consisting of an optionally substituted alkylene group, an optionally substituted polyethylene glycol group, an optionally substituted alkenylene group, an optionally substituted alkynylene group, an optionally substituted aliphatic cyclylene group, an optionally substituted aliphatic heterocyclylene group, an optionally substituted arylene group, and an optionally substituted heteroarylene group; where substituted, the substituent is selected from halogen, hydroxyl group, amino group, carboxyl group, sulfonic acid group, sulfonate group, phosphate group, and alkoxy group; B 2 is a branched group; P1 and P3 are each independently directly bonded or arbitrarily substituted polypeptide residues or glucose fragments; T1 is an arbitrarily substituted spacer group, T3 is a directly bonded or arbitrarily substituted spacer group, and at least one of B2 and T3 is substituted by a hydrophilic group; D1 and D3 are the first drug unit and the third drug unit, respectively, and are identical or different; b is a compound that is 1, 2, or 3, or its tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.
  17. In Paragraph 14, B1 and B2 are independently branched groups; P1 , P2 , and P3 are each independently and arbitrarily substituted polypeptide residues or glucose fragments; T1 is an arbitrarily substituted spacer group, T2 and T3 are each independently directly bonded or arbitrarily substituted spacer groups, and at least one of B2 , T2 , and T3 is substituted by a hydrophilic group; D1 , D2 , and D3 are the first drug unit, the second drug unit, and the third drug unit, respectively, and are identical or different; a and b are independently 1, 2 or 3 compounds, or tautomers, mesocompounds, racemic mixtures, enantiomers, diastereomers, or deuterium forms thereof, or mixtures thereof, or pharmaceutically acceptable salts, prodrugs, or solvates thereof.
  18. In any one of paragraphs 14 through 17, The above is arbitrarily substituted (Q1'), arbitrarily substituted (Q2'), arbitrarily substituted (Q3'), arbitrarily substituted (Q4'), arbitrarily substituted (Q5'), arbitrarily substituted (Q6'), arbitrarily substituted (Q7') and arbitrarily substituted A compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound form, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, selected from the group consisting of (Q8'), wherein * indicates a position connected to C1 and a wavy line indicates a position connected to an antibody.
  19. In any one of paragraphs 14 through 18, C1 is directly bonded, -( CH2 ) m1- , -( CH2 ) m1- (O- CH2 - CH2 ) m2- , -( CH2 ) m1- (O- CH2 -CH2) m2- (CH2) m3- , -( CH2 ) m1 -C(O)NH-( CH2 ) m2- , -( CH2 ) m1 - NHC(O)-( CH2 ) m2- and -(C A compound selected from C-(CH 2 ) m1 - wherein m1, m2 and m3 are independently integers from 1 to 6, and the left side of the indicated substituent is connected to Q' and the right side is connected to B 1 or P 1 , or a tautomeric isomer, mesocompound, racemic mixture, enantiomer, diastereomer, or deuterium compound form thereof, or a mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof.
  20. In any one of paragraphs 14 through 19, B 1 is directly coupled, and , Selected from a group consisting of; Preferably and and, junction point 1 is connected to C1 or Q' (when C1 is a direct bond), junction point 2 is connected to P1 , junction points *, 3, 4 and 5 are connected to P2 , L1 is selected from -( CH2 ) m1- , -( CH2 ) m1 -C(O)NH-( CH2 ) m2- , and -( CH2 ) m1 -NHC(O)-( CH2 ) m2- , L2 , L3 and L4 are independently -( CH2 ) m1- (O- CH2 - CH2 ) m2- or -( CH2 ) m1- (O- CH2 - CH2 ) m2- ( CH2 ) m3- , L5 is -( CH2 ) m1- , and L6 is -( CH2 ) m1- or -(CH 2 -CH 2 -O)m 2 , and m1, m2, m3, w and v are each independently 1, 2, 3, 4, 5 or 6; Preferably, when C1 is a direct bond, B1 is or Phosphorus, a compound, or its tautomer, mesocompound, racemic mixture, enantiomer, diastereomer, or its deuterium compound form, or a mixture thereof, or its pharmaceutically acceptable salt, prodrug, or solvate.

Description

Multidrug linker and antibody-drug conjugate The present invention relates to a multidrug (e.g., dual drug) linker used in an antibody-drug conjugate (ADC), an ADC based on said multidrug linker, a method for manufacturing said, and uses thereof. All existing antibody-conjugated drugs are single-loading drug conjugates, meaning that the same toxin linker is attached to a single antibody molecule; different ADCs differ only in the number and site of drug loading, and single-loading drug-antibody conjugates achieve targeted therapy of tumor cells by binding a single drug molecule to an antibody molecule. However, due to the heterogeneity and resistance of tumor cells, there are certain limitations to the therapeutic efficacy of single-loading drug-antibody conjugates. Over the past decade, tumor immunotherapy and antibody-conjugated drugs have achieved significant success in tumor treatment. However, while high response rates are observed only in certain specific tumors or antigen-high expression specific tumors, such as the success of HER2/3 ADCs in breast cancer, clinical application to gastrointestinal solid tumors is moderate or even poor, and these solid tumors exhibit significant heterogeneity between tumor cells and the surrounding microenvironment (TME). In other words, tumors are not merely modified masses of cells but also possess a fibrotic microenvironment. Some solid tumor types, particularly those associated with extensive fibrotic tumor stromas such as pancreatic ductal adenocarcinoma (PDAC), renal cell carcinoma (RCC), lung cancer, and colorectal cancer, respond poorly to immunotherapy. The overall fibrotic response directly and indirectly influences the therapeutic response to chemotherapy and immunotherapy; therefore, through antibody-conjugated drugs with dual loading, overall efficacy can be enhanced on the one hand, and by shifting the tumor from "cold" to "heat" on the other, the efficacy of combination therapy with immunotherapeutic drugs can be further improved. Dual toxins can reduce drug resistance because they can overcome intracellular or extracellular barriers that single toxins may face, such as the downregulation of target expression, upregulation of drug pump expression, and activation of DNA repair mechanisms. However, ADCs achieve efficient apoptosis by utilizing the high specificity and high affinity of antibodies to precisely deliver two types of synergistic chemotherapy drugs to tumor cells. Therefore, researchers have begun exploring methods to combine two or more drug molecules to form dual-loading or multi-loading drug-antibody conjugates, thereby achieving synergistic effects of various drugs and enhancing therapeutic efficacy. Currently disclosed drug molecule combinations are primarily combinations of different toxins, including combinations of the same or different mechanisms. For example, the MMAE/MMAF combination (Nat Commun 2021;12:3528; Angew Chem Int Ed Engl 2017;56:733-7; Bioconjugate Chem. 2023, 34, 4, 748-755); The combinations are DM1/MMAE (Nature Protocols 2017;12:1702-21; Pharmaceutics 2023;15:2020); PNU-159682/MMAF (Antib Ther 2019;2:71-8); MMAE/α-Amanitin (Int J Mol Sci 2018;19:2098); and MMAE/PBD (Bioorg Med Chem Lett 2018;28:3617-21). Initial efficacy experiments demonstrated that dual toxin ADC molecules can significantly enhance efficacy while simultaneously improving the apoptotic activity of ADC drugs against drug-resistant tumor cells. In addition, existing reports regarding conjugation methods and processes related to dual-toxin ADC drugs include the following: a method of simultaneously conjugating one or more drugs to a limited number of ligation sites of an antibody by serially linking MMAF and other drugs to form a dual-toxin-loaded antibody-conjugated drug molecule (PCT/CN2021/107079) and using one or more cysteine residues or cysteine derivative residues as drug linkage carriers (PCT/CN2019/112663); a method of linking existing drugs such as VC-PAB-MMAE, GGFG-DXd, and Eribulin in parallel via two-tooth linkage groups (PCT/CN2022/080942); and a method of constructing various drug-antibody-drug conjugates through “orthogonal” deprotection and drug loading among site-specific materials (PCT/US2017/066504). A method for constructing a dual payload ADC (Angew Chem Int Ed Engl 2017;56:733-7) by introducing two protected cysteine groups [Cys(SiPr)+Cys(Acm)] using a dual cysteine multiplexing carrier and combining different deprotection reactions is included, but since highly toxic catalytic reagents such as Hg(OAc) 2 must be used in the process, it increases the risk of drug safety and can simultaneously have potential adverse effects on protein structure and stability. The aforementioned prior art primarily realizes antibody conjugation of different toxins through “linear linkers” and “branched-chain linkers.” Because the solubility of toxin compositions in existing linkers is relatively low, existing dual payloads are applicable only to high-ac