Antitumor compounds and their applications

Abstract

The ligand conjugate, or its tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, comprises the structure shown in Formula I and can be used as an antitumor drug. [Formula 1] TIFF2025538619000336.tif16170

Description

Detailed Description of the Invention

[0001] [Technical field] The present application relates to the biomedical field, specifically to anti-tumor compounds and their applications.

[0002] [Background technology] Antibody drug conjugates (ADCs) conjugate monoclonal antibodies or antibody fragments to biologically active cytotoxins via stable chemical linker compounds, fully utilizing the antibody's binding specificity for normal and tumor cell surface antigens and the high cytotoxicity of the agent to exert antitumor effects. While the application of the camptothecin derivative exatecan in antibody drug conjugates (ADCs) has been reported in the literature, the art remains in need of further development of ADC drugs with better therapeutic efficacy and improved safety. However, currently available ADC drugs still suffer from issues such as poor plasma stability and off-target toxic side effects, which affect the therapeutic efficacy and safety of the products. To address these issues, the development of new stable connectors and / or connector-drug molecules is necessary.

[0003] [Summary of the Invention] [Problem to be solved by the invention] The present invention provides a ligand conjugate that is highly stable, has a favorable therapeutic effect, and is highly safe, as well as a ligand conjugate precursor, a connector, a connector precursor, and the like.

[0004] [Means for solving the problem] A first aspect of the present invention provides a ligand conjugate, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, wherein said ligand conjugate comprises the structure shown in Formula I:

[0005] [ka]

[0006] where L is an optionally substituted connector that is attached to any O, S, or N atom in the P structure; Ab is a ligand, a is a number greater than 0, and a is a decimal or integer; P is a toxin, and said P has the structure shown in Formula II:

[0007] [ka]

[0008] where n is 0 or 1, X is N or CR 0 is selected from the group consisting of R 0 is selected from the group consisting of H, D, halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, OH, NH2, N3, or NO2; R 1 is selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a C1-C8 alkyl group, a C1-C8 alkoxy group, a C1-C8 halogenated alkyl group, a C1-C8 halogenated alkoxy group, N3, NO2, NH2, NH-OH, -NR'R'', -COOR', -CONR'R'', -NHR'''NR'R'', wherein R', R'' and R''' are each independently selected from hydrogen, deuterium, an alkyl group, an aryl group, an arylalkyl group, an acyl group, an alkoxycarbonyl group and an aryloxycarbonyl group; R 2 , R 3 , R 4 , R 5 and R 6 are each independently a hydrogen atom, a deuterium atom, a halogen atom, a hydroxy group, a cyano group, NH2, NO2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 alkoxy group, a substituted or unsubstituted C1-C8 alkylthio group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m Tri(C1-C4 alkyl)silyl group, -(CH2) m(C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2, -(CH2) m S(CH2) p R 7 , -(CH2) m S(O)(CH2) p R 7 , -(CH2) m S(O)2(CH2) p R 7 , -(CH2) m NH(CH2) p R 7 , -(CH2) m NHC(O)(CH2) p R 7 , -(CH2) m OC(O)(CH2) p R 7 , -(CH2) m C(O)(CH2) p R 7 , -CH=N(OtBu), wherein m and p are each independently 0, 1, 2, 3, or 4; Or, R 2 and R 3 together with the carbon atoms to which they are attached form a substituted or unsubstituted C5-C8 carbocyclic ring or a substituted or unsubstituted 5- to 12-membered heterocyclic group; Or, R 3 and R 4 , or R 4 and R 5 together with the carbon atoms to which they are attached, form a structure selected from the group consisting of a saturated or unsaturated 5- to 12-membered carbocyclic ring that is unsubstituted or substituted by one or more Re, and a saturated or unsaturated 5- to 12-membered heterocyclic ring that is unsubstituted or substituted by one or more Re, e is a hydrogen atom, a deuterium atom, a halogen atom, a nitrile group, a nitro group, a hydroxy group, an amino group, a C1-C6 alkyl-NH-, a (C1-C6 alkyl)2N-, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C1-C6 alkoxy group, an allyl group, a benzyl group, a C6-C12 Aryl group, C1-C6 alkoxy-C1-C6 alkyl group, C1-C6 alkoxy-carbonyl group, phenoxycarbonyl group, C2-C6 alkynyl-carbonyl group, C2-C6 alkenyl-carbonyl group, C3-C6 cycloalkyl-carbonyl group, C1-C6 alkyl-sulfonyl group, phenyl group, 5-7 membered heteroaryl group, C3-C8 cycloalkyl group, 3-12 membered heterocyclic group, -(CH2) m N(R 7 )2, -(CH2) m S(CH2) p R 7 , -(CH2) m S(O)(CH2) p R 7 , -(CH2) m S(O)2(CH2) p R 7 , -(CH2) m NH(CH2) p R 7 , -(CH2) m NHC(O)(CH2) p R 7 , -(CH2) m OC(O)(CH2) p R 7 , -(CH2) m C(O)(CH2) p R 7 wherein m and p are each independently 0, 1, 2, 3, or 4, preferably 0, 1, or 2; R 7is selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a C1-C8 halogenated alkyl group, a C1-C8 deuterated alkyl group, a substituted or unsubstituted C1-C8 alkoxy group, a hydroxy group, an amino group, a cyano group, a nitro group, a mercapto group, a substituted or unsubstituted C1-C8 alkylene-OH, a substituted or unsubstituted C1-C8 alkylene-NH2, SO2Me, -OC(O)(substituted or unsubstituted C1-C4 alkyl), -C(O)(substituted or unsubstituted C1-C4 alkyl), a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5- to 7-membered heteroaryl group, a substituted or unsubstituted C3-C8 cycloalkyl group, and a substituted or unsubstituted 3- to 12-membered heterocyclic group; Unless otherwise specified, each of the above "substituted" groups means that one or more hydrogen atoms on the group have been replaced with a hydrogen atom, a deuterium atom, a halogen, a nitrile group, a nitro group, a hydroxy group, an amino group, a C1-C6 alkyl-NH-, a (C1-C6 alkyl)2N-, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C1-C6 alkoxy group, a halogenated C1-C6 alkyl group, a halogenated C2-C6 alkenyl group, a halogenated C2-C6 alkynyl group, a halogenated C1-C6 alkoxy group, an allyl group, a benzyl group, a C6-C 12 It refers to being substituted by a substituent selected from the group consisting of an aryl group, a C1-C6 alkoxy-C1-C6 alkyl group, a C1-C6 alkoxy-carbonyl group, a phenoxycarbonyl group, a C2-C6 alkynyl-carbonyl group, a C2-C6 alkenyl-carbonyl group, a C3-C6 cycloalkyl-carbonyl group, a C1-C6 alkyl-sulfonyl group, a phenyl group, a 5- to 7-membered heteroaryl group, a C3-C8 cycloalkyl group, and a 3- to 12-membered heterocyclic group.

[0009] In another preferred example, L has a structure as shown in the following formula: -L1-L2-L3-L4-L5- wherein L1 is an optionally substituted

[0010] [ka] and Rd is H, a C1-C6 alkyl group, a C1-C6 deuterated alkyl group, a C3-C8 cycloalkyl group, or a C3-C8 deuterated cycloalkyl group;

[0011] The L2 is an optionally substituted -(CHR) m1 -X1-(CH2CH2O) n3 -(CHR) m2 -C(O)-, optionally substituted -(CHR) m1 -X1-X2-(CH2CH2O) n3 -(CHR) m2 -C(O)-, optionally substituted -X-(CHROCHR) m2 -C(O)-, optionally substituted -(CHR) p 1-C(O)-, optionally substituted -(CHR) m1 -X1-(CHR) m2 -C(O)-, optionally substituted -(CHR) m1 -X1-(CHR) n3 -X2-(CHR) m2 -C(O)-, optionally substituted -X-(CHR) m1 -X2-(CHR) m2 -C(O)-, optionally substituted -(CH2CH2O) n3 is a group selected from the group consisting of —C(O)—, X1 and X2 each independently represent -O-, -C(O)-, -C(O)-NR-, or an optionally substituted C-C 10 selected from the group consisting of an aryl group, an optionally substituted 5- to 9-membered heteroaryl group, an optionally substituted 3- to 8-membered heteroalicyclic group, and an optionally substituted C3-C6 alicyclic group; Here, each R is independently H, D, (CH2) n4 OH, (CH2) n4 NH2, (CH2O) n4 (CH2CH2O) n5 H, (CHO) n4 (CH2CH2O) n5 CH3, (CH2) n4 OCH3, (CH2CH2O) n5 CH3, CH2C(O)NH(CHO) n4(CH2CH2O) n5 H, CH2C(O)NH(CHO) n4 (CH2CH2O) n5 CH3; wherein m1, m2, n3, n4, and n5 are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; and p1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, or 8; L3 is a peptide residue, and L3 is

[0012] [ka] , CH2C(O)R c and wherein R may be substituted by one or more substituents selected from the group consisting of c teeth,

[0013] [ka] wherein n1 and n2 are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;

[0014] L4 is an optionally substituted -L 4a -(NR b ) n6 -R 12 -L 4b -, where L 4a does not exist or L 4a is an arbitrarily substituted

[0015] [ka] where n6 is 0 or 1, and R 12 is a chemical bond, CH2, or CD2,

[0016] L 4b does not exist or L 4b is an arbitrarily substituted

[0017] [ka] where Ra and R b are each independently selected from the group consisting of hydrogen, optionally substituted C1-C4 alkyl groups, and optionally substituted C1-C4 deuterated alkyl groups;

[0018] The L5 is absent or optionally substituted

[0019] [ka] wherein Y is selected from the group consisting of O, S, or NH; v is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; and R 10 and R 11 are each independently selected from the group consisting of hydrogen, deuterium, optionally substituted C1-C4 alkyl groups, optionally substituted C1-C4 halogenated alkyl groups, optionally substituted C3-C6 cycloalkyl groups, and optionally substituted C4-C8 cycloalkyl groups, or R 10 and R 11 together with the atoms to which they are attached form an optionally substituted 3- to 6-membered cycloalkyl group, and R 10 and R 11 are each independently selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, an optionally substituted C1-C8 alkyl group, an optionally substituted C1-C8 halogenated alkyl group, and an optionally substituted C1-C8 deuterated alkyl group.

[0020] In another preferred embodiment, L1 is

[0021] [ka] is.

[0022] In another preferred example, X1 and X2 each independently represent -O-, -C(O)-, -C(O)-NR-, an optionally substituted phenyl group, an optionally substituted pyridyl group, an optionally substituted C3-C6 cycloalkyl group, an optionally substituted

[0023] [ka] or optionally substituted

[0024] [ka] is selected from the group consisting of:

[0025] In another preferred embodiment, L2 is an optionally substituted -(CH2) m1 -X1-(CH2CH2O) n3 -(CH2) m2 wherein X1 is —C(O)—NH—, and preferably m1 and m2 are each independently selected from 1, 2 or 3, and n3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

[0026] In another preferred embodiment, L2 is an optionally substituted -(CHR) m1 -X1-X2-(CH2CH2O) n3 -(CHR) m2 -C(O)-, where X1 is an optionally substituted

[0027] [ka] , or optionally substituted

[0028] [ka] and X2 is —C(O)—NR—, preferably, m1 and m2 are each independently selected from 0, 1 or 2, and n3 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

[0029] In another preferred embodiment, L2 is optionally substituted -X1-(CHROCHR) m2 -C(O)-, wherein X1 is an optionally substituted aryl group, an optionally substituted heteroaryl group, and preferably m2 is selected from 0, 1, 2 or 3.

[0030] In another preferred embodiment, the L2 is an optionally substituted (CHR) p1 -C(O)-, where p1 is selected from 0, 1 or 2, and R is H, (CH2) n4 OH, (CHO) n4 (CH2CH2O) n5 H, and preferably n4 and n5 are each independently selected from 0, 1, 2 or 3.

[0031] In another preferred embodiment, L2 is an optionally substituted -(CH2) m1 -X1-(CH2CH2O) n3 -(CHR) m2 wherein X1 is —C(O)—, and preferably m1 and m2 are each independently selected from 0, 1, 2 or 3, and n3 is selected from 0, 1 or 2.

[0032] In another preferred embodiment, L2 is an optionally substituted -X1-(CH2) m1 -X2-(CHR) m2 wherein X1 is an optionally substituted aryl group or an optionally substituted heteroaryl group, and X2 is -C(O)-; and preferably, m1 and m2 are each independently selected from 0, 1, or 2.

[0033] In another preferred embodiment, L2 is -(CHR) m1 -X1-(CHR) m2 -C(O)-, wherein X1 is an optionally substituted 3-8 Preferably, m1 is 0, 1 or 2, and m2 is 0.

[0034] In another preferred embodiment, L2 is an optionally substituted -(CHR) m1 -X1-(CH2CH2O) n3 -(CHR) m2 In another preferred embodiment, L2 is an optionally substituted -(CHR) m1 -X1-(CHR) n3 -X2-(CHR) m2 -C(O)-, wherein X1 is optionally substituted -C(O)-NR-, X2 is O, and preferably m1, n3, and m2 are each independently 1, 2, or 3, and R is as described above.

[0035] In another preferred embodiment, L2 is

[0036] [ka] is an optionally substituted structure selected from the group consisting of: In a preferred embodiment, the selection of the L2 structure allows the maleimide group of the resulting ligand conjugate to form an open ring structure, thereby inhibiting the Retro-Michael reaction, improving the stability of the ligand conjugate, and reducing the loss of the small molecule moiety, thereby improving the safety of the ligand conjugate drug.

[0037] In another preferred embodiment, L3 is an unsubstituted or CH2C(O)R selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine. c is a peptide residue consisting of an amino acid substituted by

[0038] In another preferred embodiment, L3 is an unsubstituted or CH2C(O)R selected from the group consisting of glycine, alanine, lysine, phenylalanine, valine, and citrulline. c is a peptide residue consisting of an amino acid substituted by

[0039] In another preferred example, L3 is -glycine-phenylalanine-glycine-(-Gly-Phe-Gly-), -glycine-glycine-phenylalanine-glycine-(-Gly-Gly-Phe-Gly-), -valine-citrulline-(-Val-Cit-), -citrulline-valine-(-Cit-Val-), -citrulline-alanine-(-Cit-Ala-), -valine-alanine-(-Val-Ala-), -valine-arginine-(-Val-Arg-), -valine-lysine-(-Val-Lys-), -valine-lysine (Ac)-(-Val-Lys(Ac)-), -lysine-valine-(-Lys-Val-), -leucine-citrulline-(-Leu-Cit-), -isoleucine-citrulline-(-Ile-Cit-), -tryptophan-citrulline-(-Trp-Cit-), -phenylalanine-lysine-(-Phe-Lys-), -phenylalanine-lysine(Ac)-(-Phe-Lys(Ac)-), -phenylalanine-citrulline-(-Phe-Cit-), -phenylalanine-alanine-(-Phe-Ala-), -phenylalanine -Arginine-(-Phe-Arg-), -Alanine-Lysine-(-Ala-Lys-), -Alanine-Alanine-(-Ala-Ala-), -Alanine-Alanine-Alanine-(-Ala-Ala-Ala-), -Alanine-Alanine-Asparagine-(-Ala-Ala-Asn-), -Alanine-Alanine-Aspartic Acid-(Ala-Ala-Asp-), -Lysine-Alanine-Alanine-Asparagine-(-Lys-Ala-Ala-Asn-), -Lysine-Alanine-Alanine-Aspartic Acid-(-Lys-Ala-Ala- Asp-), -(D)-valine-leucine-lysine-(-D-Val-Leu-Lys-), -glycine-glycine-arginine-(-Gly-Gly-Arg-), -glycine-glycine-asparagine-(-Gly-Gly-Asn-), -glycine-glycine-phenylalanine-(-Gly-Gly-Phe-), -valine-lysine-glycine-(-Val-Lys-Gly-), -glutamic acid-alanine-alanine-(-Glu-Ala-Ala-), -aspartic acid-alanine-alanine-(-Asp-Ala-Ala-),unsubstituted or CHC(O)R selected from the group consisting of -valine-lysine-glycine-glycine-(-Val-Lys-Gly-Gly-) and -lysine-alanine-asparagine-(-Lys-Ala-Asn-), c is a peptide residue substituted by

[0040] In another preferred embodiment, L3 is an unsubstituted or CH2C(O)R selected from the group consisting of: c is a structure in which

[0041] [ka] TIFF2025538619000017.tif117170TIFF2025538619000018.tif127170

[0042] In another preferred embodiment, L4 is a chemical bond, or

[0043] wherein R is an optionally substituted group selected from the group consisting of a and R b are each independently selected from the group consisting of hydrogen, an optionally substituted C1-C4 alkyl group, and an optionally substituted C1-C4 deuterated alkyl group.

[0044] [ka] In another preferred embodiment, L4 is a chemical bond, or

[0045] [ka] is an optionally substituted structure selected from the group consisting of:

[0046] In another preferred embodiment, L5 is a chemical bond or an optionally substituted

[0047] [ka] , optionally substituted

[0048] [ka] , optionally substituted

[0049] [ka] , optionally substituted

[0050] [ka] , optionally substituted

[0051] [ka] , optionally substituted

[0052] [ka] , optionally substituted

[0053] [ka] , optionally substituted

[0054] [ka] , optionally substituted

[0055] [ka] , optionally substituted

[0056] [ka] , optionally substituted

[0057] [ka] , optionally substituted

[0058] [ka] and optionally substituted

[0059] [ka] is a structure selected from the group consisting of:

[0060] In another preferred embodiment, the compound of formula II is selected from the group consisting of:

[0061] [ka]

[0062] In another preferred embodiment, the compound of formula II is selected from the group consisting of:

[0063] [ka]

[0064] In another preferred embodiment, the compound of formula II is selected from the group consisting of:

[0065] [ka]

[0066] In another preferred embodiment, the R 4 is a hydrogen atom, a deuterium atom, a halogen atom, a hydroxy group, a cyano group, NH2, NO2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted a substituted C1-C8 alkoxy group, a substituted or unsubstituted C1-C8 alkylthio group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C8 cycloalkyl), -(CH2) m (3-12 membered heterocyclic group), -(CH2) m N(R 7 )2, -(CH2) m S(O)(CH2) p R 7 , -(CH2) m S(O)2(CH2) p R 7 , -(CH2) m NH(CH2) p R 7 wherein m and p are each independently 0, 1, or 2; 7 is defined as above, R 5 is selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen atom, NH, OH, a substituted or unsubstituted C-C alkyl group, and a substituted or unsubstituted C-C alkoxy group; Or, R 4 and R 5 together with the carbon atoms to which they are attached, are unsubstituted or one or more R e saturated or unsaturated 5- to 6-membered carbocyclic ring, unsubstituted or substituted by one or more R e and forming a structure selected from the group consisting of saturated or unsaturated 5- to 6-membered heterocycles substituted by e The definition of is as described above.

[0067] In another preferred embodiment, R 2 and R 3 are each independently a hydrogen atom, a deuterium atom, a halogen atom, NH, a substituted or unsubstituted C-C alkyl group, a substituted or unsubstituted C-C deuterated alkyl group, -(CH) m (C3-C6 cycloalkyl), -(CH2) m (3- to 6-membered heterocyclic group), -(CH2) m N(R 7 )2, -(CH2)m OC(O)R 7 wherein m is 0, 1, 2, 3, or 4; and R 7 is defined as above, Or, R 2 and R 3 together with the carbon atoms to which they are attached, are unsubstituted or one or more R e saturated or unsaturated 5- to 6-membered ring, unsubstituted or substituted by one or more R e and forming a structure selected from the group consisting of saturated or unsaturated 5- to 6-membered heterocycles substituted by e The definition of is as described above.

[0068] In another preferred embodiment, R 1 and R 6 are each independently a hydrogen atom. In another preferred embodiment, the R 4 and R 5 are each independently selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxy group, NH2, and a substituted or unsubstituted C1-C4 alkyl group; Or, R 4 and R 5 together with the carbon atoms to which they are attached, are unsubstituted or one or more R e and forming an oxa 5-6 membered heterocycle substituted by e The definition of is as described above.

[0069] In another preferred embodiment, R 2 represents a deuterium atom, a halogen, NH2, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3- to 6-membered heterocyclic group), -(CH2) m N(R 7 )2, -(CH2) m OC(O)R 7wherein m is 0, 1, 2, 3, or 4; R 3 are respectively a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted C1-C8 deuterated alkyl group, -(CH2) m (C3-C6 cycloalkyl), -(CH2) m (3- to 6-membered heterocyclic group), -(CH2) m N(R 7 )2, -(CH2) m OC(O)R 7 wherein m is 0, 1, 2, 3, or 4; Or, R 2 and R 3 together with the carbon atoms to which they are attached, are unsubstituted or one or more R e saturated or unsaturated 5- to 6-membered ring, unsubstituted or substituted by one or more R e and forming a structure selected from the group consisting of saturated or unsaturated 5- to 6-membered heterocycles substituted by R 4 is selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a hydroxy group, a cyano group, NH2, NO2, a substituted or unsubstituted C1-C8 alkyl group, and a substituted or unsubstituted C1-C8 alkoxy group; R 5 is a hydrogen atom, a deuterium atom, a halogen atom, a substituted or unsubstituted C1-C8 alkyl group is selected from the group consisting of Or, R 4 and R 5 together with the carbon atom to which they are attached form a group selected from the group consisting of —OCHO— or —O(CH)O—, unsubstituted or substituted by one or more Re; R 7 is selected from the group consisting of a hydrogen atom, a deuterium atom, a halogen, a substituted or unsubstituted C1-C8 alkyl group, a hydroxy group, an amino group, a cyano group, a nitro group, and a mercapto group; where R e The definition of is as described above.

[0070] In another preferred embodiment, the compound of formula II is selected from the group consisting of:

[0071] [ka] TIFF2025538619000038.tif153170TIFF2025538619000039.tif220170TIFF2025538619000040.t if185170TIFF2025538619000041.tif203170TIFF2025538619000042.tif218170TIFF2025538619 000043.tif191170TIFF2025538619000044.tif204170TIFF2025538619000045.tif209170TIFF20 25538619000046.tif171170TIFF2025538619000047.tif215170TIFF2025538619000048.tif77170

[0072] In a preferred embodiment, the ligand complex is selected from the group consisting of:

[0073] [ka] TIFF2025538619000050.tif117170TIFF2025538619000051.tif207170TIFF2025538619000052.tif198170TIFF2025538619000053.tif121170In the structure shown above, "-L1-" is

[0074] [ka] The structure is selected from the following:

[0075] In a preferred embodiment, the ligand conjugate is selected from the group consisting of:

[0076] [ka] TIFF2025538619000056.tif197170TIFF2025538619000057.tif79170

[0077] In the structure shown above, "-L1-" means

[0078] [ka] and the like, preferably

[0079] [ka] and more preferably

[0080] [ka] is.

[0081] In another preferred embodiment, the ligand conjugate comprises the structure shown in formula (Ia):

[0082] [ka]

[0083] where R 4 , R 5 and L is as defined in the first aspect of the present invention, a is a number greater than 0, and a is a decimal number or an integer. In another preferred embodiment, the ligand complex has a structure selected from the group consisting of:

[0084] [ka] TIFF2025538619000063.tif179170TIFF2025538619000064.tif192170TIFF2025538619000065.tif138170

[0085] Here, a is a number greater than 0, and a is a decimal number or an integer. In another preferred example, a is an integer or decimal number other than zero from 0 to 8, preferably an integer or decimal number from 1 to 8, more preferably 2 to 8, and may be an integer or a decimal number, and most preferably 3 to 8, and may be an integer such as 4.0 or a decimal number such as 3.9.

[0086] In another preferred embodiment, the Ab is an antibody or an antigen-binding fragment thereof. In another preferred embodiment, the antibody is selected from the group consisting of a murine antibody, a chimeric antibody, a humanized antibody, and a fully human antibody.

[0087] In another preferred embodiment, the antibody is a monoclonal antibody. In another preferred embodiment, the antigen-binding fragment is selected from the group consisting of Fab, Fab', Fv fragment, F(ab')2, F(ab)2, scFv, di-scFv, VHH, and dAb.

[0088] In another preferred embodiment, the antibody is a Her-2-specific antibody (e.g., Trastuzumab), a Trop2-specific antibody (e.g., humanized RS7 or Sacituzumab in US7238785B2), a PSMA-specific antibody (e.g., PSMA-specific monoclonal antibody AB-PG1-XG1-006 in WO2003034903A2), an FR-α-specific antibody (e.g., humanized LK26 antibody (Farletuzumab, MORAb-003) in US5952484), a B7H3 antibody (e.g., antibody P7-C05-H4L3 in WO2021244590A1), or an I GF-1R specific antibodies.

[0089] In another preferred embodiment, the antibody is an IGF-1R-specific antibody. In another preferred embodiment, the IGF-1R comprises an IGF-1R derived from a primate.

[0090] In another preferred embodiment, the antibody is an IgG1 or a mutant thereof. In another preferred embodiment, the antibody comprises an HCDR3, wherein the HCDR3 is It comprises the amino acid sequence shown in ID NO:3.

[0091] In another preferred embodiment, the antibody comprises an HCDR2, wherein the HCDR2 is It comprises the amino acid sequence shown in ID NO:2. In another preferred embodiment, the antibody comprises an HCDR1, wherein the HCDR1 is It comprises the amino acid sequence shown in ID NO:1.

[0092] In another preferred example, the antibody comprises a heavy chain variable region VH, wherein the VH comprises the HCDR1, HCDR2 and HCDR3, wherein the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:3, the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:2, and the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:1.

[0093] In another preferred embodiment, the antibody comprises a heavy chain variable region VH, wherein the VH is It comprises the amino acid sequence shown in ID NO:4. In another preferred embodiment, the antibody has the full-length sequence as shown in SEQ ID No. 5.

[0094] In another preferred example, the antibody is a variant of any of the above antibodies, and the variant comprises the CDR region. In another preferred example, the variant is a sequence formed by substituting, deleting, and / or adding one or more amino acids to the amino acid sequence of the antibody (e.g., substituting, deleting, and / or inserting 1 to 30, 1 to 20, or 1 to 10, e.g., 1, 2, 3, 4, 5, 6, 7, 8, or 9 amino acids).

[0095] In another preferred example, the variant is a homolog of the amino acid sequence of the antibody, and the homolog may be an amino acid sequence having at least about 85% sequence identity (e.g., at least about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more) with the amino acid sequence of the CDR.

[0096] In another preferred embodiment, the CDRs are determined by the Kabat numbering system. A second aspect of the present invention provides a ligand conjugate precursor, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, wherein said ligand conjugate precursor comprises the structure shown in formula IA: Formula (IA): L A -P where L is L 1A -L2-L3-L4-L5-, where L 1A teeth,

[0097] [ka] where R d , L2, L3, L4, L5 and P are as defined in the first aspect of the present invention.

[0098] In another preferred embodiment, the ligand conjugate precursor is selected from the following structures:

[0099] [ka] TIFF2025538619000068.tif196170TIFF2025538619000069.tif201170TIFF2025538619000070.tif209170TIFF2025538619000071.tif213170

[0100] In the structure shown above, L 1A teeth,

[0101] [ka] is selected from.

[0102] A third aspect of the present invention provides a connector as shown in formula (L), which links a Drug unit to a Ligand to form a Ligand-Drug conjugate: Formula (L): L1-L2-L3-L4-L5 Here, the definitions of L1, L2, L3, L4, and L5 are as described in the first aspect of the present invention.

[0103] In another preferred embodiment, the connector is selected from the group consisting of:

[0104] [ka] TIFF2025538619000074.tif203170TIFF2025538619000075.tif187170TIFF2025538619000076.tif195170TIFF2025538619000077.tif213170

[0105] In another preferred embodiment, the connector is linked to a ligand via an L1 segment and to P1 via an L5 segment to form a ligand-drug conjugate, and P1 is selected from the group consisting of glycopeptide antibiotics such as bleomycin or bleomycin; DNA topoisomerase inhibitors such as topoisomerase I inhibitors (e.g., camptothecin, hydroxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, isotecan, topotecan, belotecan, rubitecan, DXd, etc.), topoisomerase II inhibitors (e.g., actinomycin D, doxorubicin, duocarmycin, daunorubicin, mitoxantrone, podophyllotoxin, or etoposide); and methotrexate. drugs that interfere with DNA synthesis, such as methadone, 5-fluorouracil, cytarabine, gemcitabine, mercaptopurine, pentostatin, fludarabine, cladribine, or nelarabine; drugs that act on structural proteins, such as tubulin inhibitors, vinca alkaloids, vincristine, vinblastine, paclitaxel, docetaxel, or cabazitaxel; inhibitors of tumor signaling pathways, such as serine / threonine kinase inhibitors, tyrosine kinase inhibitors, aspartate kinase inhibitors, or histidine kinase inhibitors; proteasome inhibitors; histone deacetylase inhibitors; tumor angiogenesis inhibitors; cell cycle protein inhibitors; maytansine derivatives (e.g., DM1, DM4, etc.); calicheamicin derivatives; auristatin derivatives (e.g., monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), auristatin E, auristatin F, etc.); pyrrolobenzodiazepine dimers dimers (PBD) derivatives; Amanita mushroom derivatives (such as α-amanitin); anthracyclines; ducalcins; eribulin; melphalan; mitomycin C; chlorambucil; TLR agonists; STING agonists; glucocorticoids, as well as groups formed by dehydrogenation of other active substances that inhibit tumor cell growth and promote tumor cell apoptosis or necrosis.

[0106] A fourth aspect of the present invention provides a connector precursor as shown in formula (L-1), which is used to link a drug unit to a ligand to obtain a ligand-drug conjugate formed thereon, which is used to link a Drug unit to a Ligand to obtain a Ligand-Drug Conjugate; Formula (L-1):L A -R g where R g is H, OH, O(C1-C6 alkyl), where L A is as defined in the third aspect of the present invention.

[0107] In another preferred embodiment, the connector precursor is selected from the group consisting of:

[0108] [ka] TIFF2025538619000079.tif206170TIFF2025538619000080.tif201170TIFF2025538619000081.tif155170

[0109] At the same time, the present invention further provides a synthesis scheme of another preferred example of (L-1), which is described as follows:

[0110] [ka]

[0111] Scheme 1: Pg1 is

[0112] [ka] are selected from,

[0113] Pg2 is selected from Boc, Fmoc, Cbz, etc. Pg3 is selected from Boc, Fmoc, Cbz, etc. Scheme 2:

[0114] [ka]

[0115] Pg1 is

[0116] [ka] are selected from,

[0117] Pg2 is selected from Boc, Fmoc, Cbz, etc. Pg4 is Me, Et, i Pr, Allyl, t It is selected from Bu, Bn, 4-methylbenzyl, 4-methoxybenzyl, 2,4-Dimethoxybenzyl, 2,6-Dimethoxybenzyl, trimethylsilyl, tert-butyldimethylsilyl, pentafluorophenyl, and the like.

[0118] A fifth aspect of the present invention provides a pharmaceutical composition comprising a ligand conjugate according to the first aspect of the invention, or a tautomeric, meso-, racemic, enantiomeric, diastereomeric or mixture thereof form thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and optionally a pharmaceutically acceptable carrier.

[0119] A sixth aspect of the present invention provides the use of a ligand conjugate according to the first aspect of the present invention, or a tautomer, meso form, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and / or a pharmaceutical composition according to the fifth aspect of the present invention, in the preparation of a medicament for the treatment and / or prevention of a disease or condition associated with target point expression and / or abnormal expression of said ligand.

[0120] In another preferred embodiment, the disease or condition associated with expression and / or abnormal expression of the target point of the ligand is a tumor / cancer, an autoimmune disease or an infectious disease, and preferably, the tumor / cancer is a tumor / cancer with high, medium or low expression of the target point of the ligand.

[0121] In another preferred embodiment, the tumor is selected from tumors associated with expression of the target points Her-2, Trop2, PSMA, FR-α, B7H3, or IGF-1R.

[0122] In another preferred embodiment, the tumor comprises a solid tumor and / or a hematological tumor. In another preferred embodiment, the tumor is a tumor associated with IGF-1R target point expression.

[0123] In another preferred embodiment, the tumor is selected from the group consisting of breast cancer, ovarian cancer, cervical cancer, endometrial cancer, urothelial cancer, lung cancer, prostate cancer, colorectal cancer, gastric cancer, esophageal cancer, bladder cancer, kidney cancer, pancreatic cancer, thyroid cancer and head and neck cancer, astrocytoma, basal cell carcinoma or squamous cell carcinoma, brain tumor, neuroblastoma, glioblastoma, cellular sarcoma, bladder cancer, colorectal cancer, colon cancer, chondrosarcoma, kidney cancer, choriocarcinoma, leukemia, multiple myeloma, Ewing's sarcoma, gastrointestinal cancer, head and neck cancer, liver cancer, glioma, hepatocellular carcinoma, leiomyoma, melanoma, non-small cell lung cancer, nervous system cancer, pancreatic cancer, renal cell carcinoma, rhabdomyosarcoma, small cell lung cancer, thymoma, thyroid cancer, testicular cancer and osteosarcoma.

[0124] Another aspect of the present invention provides a method for preparing a connector precursor as shown in formula (L-1): The method includes the following steps:

[0125] [ka] By reacting with

[0126] [ka] Obtained,

[0127] where L 1A teeth,

[0128] [ka] and preferably, L 1A teeth,

[0129] [ka] and more preferably, L 1A teeth,

[0130] [ka] is.

[0131] L2 is as described in any of the above embodiments, preferably L2 is

[0132] [ka] and more preferably, L2 is selected from the group consisting of:

[0133] [ka] More preferably, L2 is selected from

[0134] [ka] is selected from.

[0135] Preferably,

[0136] [ka] is selected from amino acids or dipeptide fragments, preferably

[0137] [ka] is glycine

[0138] [ka] or a glycine-glycine fragment

[0139] [ka] and preferably

[0140] [ka] is a glycine-glycine fragment

[0141] [ka] and

[0142] Preferably,

[0143] [ka] is selected from amino acids or dipeptide fragments, preferably

[0144] [ka] is glycine

[0145] [ka] or a glycine-glycine fragment

[0146] [ka] and preferably

[0147] [ka] is a glycine-glycine fragment

[0148] [ka] and

[0149] Preferably,

[0150] [ka] is a dipeptide or tripeptide fragment, preferably

[0151] [ka] is the glycine-phenylalanine-glycine fragment

[0152] [ka] , phenylalanine-glycine fragment

[0153] [ka] and preferably

[0154] [ka] is a phenylalanine-glycine fragment

[0155] [ka] and

[0156] Preferably,

[0157] [ka] is a dipeptide or tripeptide fragment, preferably

[0158] [ka] is the glycine-phenylalanine-glycine fragment

[0159] [ka] , phenylalanine-glycine fragment

[0160] [ka] and preferably

[0161] [ka] is a phenylalanine-glycine fragment

[0162] [ka] and

[0163] Pg1 is

[0164] [ka] Preferably, Pg1 is selected from

[0165] [ka] Preferably, Pg1 is selected from

[0166] [ka] Preferably, Pg1 is selected from

[0167] [ka] Preferably, Pg1 is selected from

[0168] [ka] is selected from

[0169] Preferably, the

[0170] [ka] is prepared by the following method:

[0171] (1)L 1A -L2-OPg5

[0172] [ka] React with

[0173] [ka] to obtain, preferably reacting with or without a base, preferably reacting without the participation of a base, preferably said base being selected from triethylamine, N,N-diisopropylethylamine, 2,4,6-trimethylpyridine, DMAP, DBU, NMM, NaHCO3, preferably said base being NaHCO3;

[0174] Pg5 is

[0175] [ka] Preferably, Pg5 is selected from

[0176] [ka] Preferably, Pg5 is selected from

[0177] [ka] Preferably, Pg5 is selected from

[0178] [ka] Preferably, Pg5 is selected from

[0179] [ka] is selected from

[0180] (2) Pg1OH and

[0181] [ka] By reacting with

[0182] [ka] Obtained,

[0183] Preferably, the Pg1OH is

[0184] [ka] Preferably, said Pg1OH is selected from:

[0185] [ka] Preferably, said Pg1OH is selected from:

[0186] [ka] Preferably, said Pg1OH is selected from:

[0187] [ka] Preferably, said Pg1OH is selected from:

[0188] [ka] is selected from

[0189] Preferably, the

[0190] [ka] is prepared by the following method:

[0191] [ka]

[0192] (i)

[0193] [ka] By the reaction of

[0194] [ka] Obtained,

[0195] (ii)

[0196] [ka] and

[0197] [ka] By reacting with

[0198] [ka] Obtained,

[0199] The method is carried out by the following method (iii) or (iv):

[0200] [ka] and further comprising the step of obtaining

[0201] (iii) First

[0202] [ka] Deprotection of Pg4 from

[0203] [ka] Then we get

[0204] [ka] Deprotection of Pg2 from

[0205] [ka] Obtained,

[0206] Preferably, the deprotection is carried out in the presence of a base, preferably the base is an organic base, more preferably the base is DBU; preferably, the deprotection is carried out in the presence of a reducing agent, preferably the reducing agent is H; preferably, the deprotection is carried out in the presence of a catalyst, preferably the catalyst is palladium on carbon Pd / C; (iv) First

[0207] [ka] Deprotection of Pg2 from

[0208] [ka] and then

[0209] [ka] Deprotection of Pg4 from

[0210] [ka] Obtained,

[0211] Preferably, the deprotection is carried out in the presence of a base, preferably the base is an organic base, more preferably the base is DBU; preferably, the deprotection is carried out in the presence of a reducing agent, preferably the reducing agent is H; preferably, the deprotection is carried out in the presence of a catalyst, preferably the catalyst is palladium on carbon Pd / C; wherein Pg2 is selected from Boc, Fmoc, and Cbz; Pg4 is Me, Et, i Pr, Allyl,t Bu, Bn, 4-methylbenzyl group, 4-methoxybenzyl group, 2,4-dimethoxybenzyl group, 2,6-dimethoxybenzyl group group, a trimethylsilyl group, a t-butyldimethylsilyl group, and a pentafluorophenyl group.

[0212] In another preferred embodiment, the connector precursor comprises:

[0213] [ka] is.

[0214] Another aspect of the present invention provides a method for preparing a compound according to formula 4:

[0215] [ka]

[0216] The method includes the steps of: The compound represented by the following formula 15h is subjected to a condensation reaction with the compound represented by the following formula 1d in an inert solvent to obtain the compound represented by the formula 4.

[0217] [ka]

[0218] In another preferred embodiment, the inert solvent is selected from the group consisting of dichloromethane, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, HMPA, or a combination thereof, preferably N,N-dimethylformamide; In another preferred embodiment, the reaction is carried out in the presence of a condensing agent, and more preferably, the condensing agent is HATU, HBTU, TBTU, EDCI, HOAt, HOBt, CDI, TC FH, TFFH, DCC, DIC, BOP, AOP, PyAOP, BrOP, PyClOP, PyBrOP, DMTMM, or a combination thereof, more preferably, the condensing agent is HATU; In another preferred example, the reaction is carried out in the presence of a base, and more preferably, the base is selected from the group consisting of TEA, DIPEA, DBU, DMAP, pyridine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, imidazole, N-methylimidazole, or a combination thereof, and more preferably, the base is 2,4,6-trimethylpyridine.

[0219] Another aspect of the present invention provides a method for preparing a compound according to formula 15h:

[0220] [ka]

[0221] The method includes the steps of:

[0222] [ka]

[0223] (1) reacting a compound represented by the following formula 15f with a compound represented by the following formula 8e in an inert solvent to obtain a compound represented by the following formula 15g;

[0224] [ka]

[0225] In another preferred example, the inert solvent is selected from the group consisting of acetonitrile, water, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or a combination thereof, more preferably acetonitrile, water, or a combination thereof, even more preferably a combination of acetonitrile and water in a volume ratio of 10:1 to 1:10, more preferably a combination of acetonitrile and water in a volume ratio of 2:1 to 4:1, more preferably a combination of acetonitrile and water in a volume ratio of 3:1, In another preferred embodiment, the reaction can be carried out in the presence of a base, and preferably, the base is selected from TEA, DIPEA, NMM, sodium bicarbonate, DBU, or a combination thereof, and more preferably, the base is DIPEA; (2) In an inert solvent, the compound represented by the following formula 15g is subjected to a silicon-based removal reagent to remove the protecting group, thereby obtaining the compound represented by the formula 15h:

[0226] [ka]

[0227] In another preferred example, the silicon-based removal reagent is selected from the group consisting of an acid and a fluoride reagent (fluorine-containing reagent), and preferably, the silicon-based removal reagent is selected from hydrogen fluoride, an aqueous hydrogen fluoride solution, triethylamine trihydrofluoride, pyridine hydrofluoride, formic acid, acetic acid, trifluoroacetic acid, dichloroacetic acid, methanesulfonic acid, and trifluoromethanesulfonic acid, and preferably, the silicon-based removal reagent is formic acid; In another preferred example, the inert solvent is selected from the group consisting of tetrahydrofuran, N,N-dimethylformamide, acetonitrile, water, or a combination thereof, more preferably acetonitrile, water, or a combination thereof, more preferably a volume ratio of acetonitrile to water of 10:1 to 1:10, more preferably a volume ratio of acetonitrile:water of 1:4 to 4:1, and more preferably a volume ratio of acetonitrile:water of 1:1.5.

[0228] In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 8e:

[0229] [ka]

[0230] The method includes the steps of:

[0231] [ka]

[0232] (1) removing the Fmoc protecting group from a compound represented by the following formula 8c under basic conditions in an inert solvent to obtain a compound represented by formula 15b;

[0233] [ka]

[0234] In another preferred example, the inert solvent is selected from dichloromethane, tetrahydrofuran, 2-methyltetrahydrofuran, or a combination thereof, more preferably, the inert solvent is dichloromethane; In another preferred example, the base is an organic base, more preferably, the base is dimethylamine, diethylamine, DBU, or piperidine, more preferably, the base is DBU.

[0235] (2) removing the benzyl protecting group from a compound represented by the following formula 15b in an inert solvent to obtain a compound represented by the following formula 8e;

[0236] [ka]

[0237] In another preferred example, the inert solvent is selected from tetrahydrofuran, water, or a combination thereof, more preferably, the inert solvent is water, and more preferably, the inert solvent is a combination of tetrahydrofuran and water in a volume ratio of 1:5 to 5:1.

[0238] In another preferred example, the deprotection is carried out in the presence of a base, preferably, the base is an organic base, more preferably, the base is DBU; In another preferred embodiment, the deprotection is carried out in the presence of a reducing agent, more preferably, the reducing agent is hydrogen gas H2; more preferably, the deprotection is carried out in the presence of a catalyst, more preferably, the catalyst is palladium on carbon Pd / C; In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 8c:

[0239] [ka]

[0240] The method includes the steps of:

[0241] [ka]

[0242] Reacting the compound of formula 8b with benzyl glycolic acid in an inert solvent to obtain the compound of formula 8c; In another preferred example, the inert solvent is selected from dichloromethane, tetrahydrofuran, 2-methyltetrahydrofuran, dioxane, or a combination thereof, and more preferably, the inert solvent is tetrahydrofuran.

[0243] In another preferred example, the reaction is carried out in the presence of an acid or a base, more preferably the acid is p-toluenesulfonic acid, p-toluenesulfonic acid monohydrate, more preferably the base is lithium hydroxide, lithium hydroxide monohydrate, potassium t-butoxide, sodium t-butoxide, sodium hydroxide, more preferably the base is lithium hydroxide, more preferably the base is lithium hydroxide monohydrate, In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 8b:

[0244] [ka]

[0245] The method includes the steps of:

[0246] [ka]

[0247] (1) reacting a compound represented by the following formula 15a with a diglycine peptide in an inert solvent to obtain a compound represented by the following formula 8a;

[0248] [ka]

[0249] In another preferred embodiment, the reaction is carried out in the presence of a base, more preferably, the base is sodium bicarbonate; In another preferred example, the inert solvent is selected from ethylene glycol dimethyl ether, water, or a combination thereof, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water in a volume ratio of 1:5 to 5:1, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water in a volume ratio of 1:2 to 2:1, (2) reacting a compound represented by the following formula 8a in an inert solvent to obtain a compound represented by the following formula 8b:

[0250] [ka]

[0251] In another preferred example, the inert solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, and HMPA, and more preferably, the inert solvent is N,N-dimethylformamide.

[0252] In another preferred embodiment, the reaction is carried out in the presence of lead tetraacetate, In another preferred embodiment, the reaction is carried out in the presence of copper acetate, In another preferred embodiment, the reaction is carried out in the presence of acetic acid, In another preferred embodiment, the reaction is carried out in the simultaneous presence of acetic acid, lead tetraacetate, and copper acetate; In another preferred embodiment, the reaction is carried out in the simultaneous presence of acetic acid and lead tetraacetate, In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 15f:

[0253] [ka]

[0254] The method includes the steps of:

[0255] [ka]

[0256] (1) In an inert solvent, a compound represented by the following formula 15c is subjected to a condensation reaction with tetrafluorophenol to obtain a compound represented by the following formula 15d:

[0257] [ka]

[0258] In another preferred embodiment, the inert solvent is selected from dichloromethane, tetrahydrofuran, and 2-methyltetrahydrofuran, and more preferably, the inert solvent is dichloromethane; In another preferred example, the reaction is carried out in the presence of a condensing agent, more preferably the condensing agent is selected from the group consisting of HATU, HBTU, TBTU, EDCI, HOAt, HOBt, CDI, TCFH, TFFH, DCC, DIC, BOP, AOP, PyAOP, BrOP, PyClOP, PyBrOP, DMTMM, or a combination thereof, more preferably the condensing agent is DCC.

[0259] (2) reacting a compound represented by the following formula 15d with a diglycine peptide in an inert solvent to obtain a compound represented by the following formula 15e:

[0260] [ka]

[0261] In another preferred example, the inert solvent is selected from ethylene glycol dimethyl ether, water, or a combination thereof, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water in a volume ratio of 1:5 to 5:1, more preferably the inert solvent is a combination of ethylene glycol dimethyl ether and water in a volume ratio of 1:2 to 2:1, In another preferred embodiment, the reaction is carried out with or without a reaction base, and more preferably, the base is selected from triethylamine, N,N-diisopropylethylamine, 2,4,6-trimethylpyridine, DMAP, DBU, NMM, NaHCO3, and more preferably the base is NaHCO3, (3) In an inert solvent, a compound represented by the following formula 15e is subjected to a condensation reaction with tetrafluorophenol to obtain a compound represented by the following formula 15f:

[0262] [ka]

[0263] In another preferred embodiment, the inert solvent is selected from dichloromethane, tetrahydrofuran, and 2-methyltetrahydrofuran, and more preferably, the inert solvent is dichloromethane; In another preferred example, the reaction is carried out in the presence of a condensing agent, more preferably the condensing agent is selected from the group consisting of HATU, HBTU, TBTU, EDCI, HOAt, HOBt, CDI, TCFH, TFFH, DCC, DIC, BOP, AOP, PyAOP, BrOP, PyClOP, PyBrOP, DMTMM, or a combination thereof, more preferably the condensing agent is DCC.

[0264] In another preferred example, the reaction is carried out in the presence or absence of a base, more preferably the reaction does not require the participation of a base; In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 15c:

[0265] [ka]

[0266] The method includes the steps of:

[0267] [ka]

[0268] (1) reacting a compound represented by the following formula 4a with a silicon-based reagent in an inert solvent to obtain a compound represented by the following formula 4b:

[0269] [ka]

[0270] In another preferred embodiment, the inert solvent is selected from the group consisting of acetonitrile, DMF, DMA, THF, and dichloromethane, and more preferably, the inert solvent is acetonitrile; In another preferred embodiment, the reaction is carried out in the presence of a base, and more preferably, the base is selected from DBU, imidazole, DIPEA, and TEA, and more preferably, the base is DBU; In another preferred embodiment, the silicon-based reagent is TBSCl or TBSOTf, and more preferably, the silicon-based reagent is TBSCl; (2) In an inert solvent, a compound represented by the following formula 4b is reacted with N-methoxycarbonylmaleimide in the presence of a base to obtain a compound represented by formula 15c:

[0271] [ka]

[0272] In another preferred example, the inert solvent is selected from the group consisting of water, THF, DMF, or a combination thereof, and more preferably, the volume ratio of the inert tetrahydrofuran to water is 1:1; In another preferred example, the base is selected from the group consisting of sodium bicarbonate, sodium carbonate, triethylamine, and diisopropylethylamine, and more preferably, the base is sodium bicarbonate.

[0273] In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 1d:

[0274] [ka]

[0275] The method includes the steps of:

[0276] [ka]

[0277] (1) In an inert solvent, a compound represented by the following formula 14g and a compound represented by the following formula 14d are subjected to a cyclocondensation reaction in the presence of an additive to obtain a compound represented by the following formula 14k,

[0278] [ka]

[0279] In another preferred example, the inert solvent is preferably selected from toluene, acetic acid, chlorobenzene, or a combination thereof, more preferably a combination of toluene and acetic acid, more preferably a combination of toluene and acetic acid in a volume ratio of 1:5 to 5:1, more preferably a combination of toluene and acetic acid in a volume ratio of 1:2 to 2:1, more preferably a combination of toluene and acetic acid in a volume ratio of 1:1, In another preferred example, the reaction can be carried out in the presence of an additive, and more preferably, the additive can be selected from the group consisting of p-toluenesulfonic acid, p-toluenesulfonic acid monohydrate, and p-toluenesulfonic acid pyridine salt, and more preferably, the additive is p-toluenesulfonic acid pyridine salt.

[0280] (2) removing the Fmoc protecting group from a compound represented by the following formula 14k under organic base conditions in an inert solvent to obtain a mixture of formula 1d and formula 1d';

[0281] [ka]

[0282] In another preferred example, the organic base is selected from a secondary amine and a tertiary amine, more preferably, the organic base is selected from piperidine, dimethylamine, and diethylamine, more preferably, the organic base is piperidine; In another preferred example, the inert solvent is selected from tetrahydrofuran and 2-methyltetrahydrofuran, and more preferably, the inert solvent is tetrahydrofuran.

[0283] (3) The mixture of 1d and 1d' obtained is subjected to chiral resolution to obtain 1d. In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 14g:

[0284] [ka]

[0285] The method includes the steps of:

[0286] [ka]

[0287] (1) In an inert solvent, the compound of formula 14e is deacetylated to give the compound of formula 14f. In another preferred example, the inert solvent is selected from no solvent, water, tetrahydrofuran, or a combination thereof, more preferably the inert solvent is water, more preferably the inert solvent is no solvent; In another preferred example, the reaction is carried out in the presence of an acid, more preferably, the acid is hydrochloric acid, more preferably, the concentration of the hydrochloric acid is 3 to 9 N, more preferably, the concentration of the hydrochloric acid is 6 N; (2) reacting the compound of formula 14f with FmocCl in the presence of a base to obtain the compound of formula 14g; In another preferred example, the base is selected from an organic base and an inorganic base, more preferably, the base is selected from triethylamine, diisopropylethylamine, DBU, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, more preferably, the base is potassium carbonate; In another preferred example, the inert solvent is selected from acetonitrile, DMF, DMA, dichloromethane, water, tetrahydrofuran, or a combination thereof, more preferably, the inert solvent is a combination of water and tetrahydrofuran in a volume ratio of 1:5 to 5:1, more preferably, the inert solvent is a combination of water and tetrahydrofuran in a volume ratio of 3:4; In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 14b:

[0288] [ka]

[0289] The method includes the steps of:

[0290] [ka]

[0291] Silicon-based protection is carried out by reacting the compound of formula 14a with two different silicon-based reagents in an inert solvent in the presence of a base to give the compound of formula 14b; In another preferred embodiment, the silicon-based reagent is TESCl, TESOTf, TIPSCl, or TIPSOTf, and more preferably, the silicon-based reagent is selected from TESCl and TIPSCl; In another preferred example, the inert solvent is selected from N,N-dimethylformamide, N,N-dimethylacetamide, HPMA, tetrahydrofuran, and dichloromethane; more preferably, the inert solvent is selected from N,N-dimethylformamide; In another preferred example, the base is preferably selected from TEA, DIPEA, NMM, pyridine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, imidazole, DMAP, more preferably the base is imidazole.

[0292] In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 14c:

[0293] [ka]

[0294] The method includes the steps of:

[0295] [ka]

[0296] Reacting the compound of formula 14b with Lawesson's reagent in an inert solvent to obtain the compound of formula 14c; In another preferred embodiment, the inert solvent is selected from tetrahydrofuran, dioxane, 2-methyltetrahydrofuran, and toluene, and more preferably, the inert solvent is toluene; In another preferred example, the reaction can be carried out with or without a base, and the base is preferably selected from triethylamine, DIPEA, NMM, pyridine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, DMAP, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, potassium t-butoxide, sodium t-butoxide, more preferably selected from sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, more preferably sodium carbonate.

[0297] In another aspect of the present invention, the present invention further provides a method for preparing a compound according to formula 14d:

[0298] [ka]

[0299] The method includes the steps of:

[0300] [ka]

[0301] The TIPS and TES silicon-based protecting groups are removed from the compound of formula 14c under the action of a silicon-based removing reagent in an inert solvent to give the compound of formula 14d.

[0302] In another preferred example, the silicon-based removal reagent is selected from the group consisting of an acid and a fluoride reagent (fluorine-containing reagent), more preferably, the silicon-based removal reagent is selected from formic acid, acetic acid, trifluoroacetic acid, methanesulfonic acid, trifluoromethanesulfonic acid, dichloroacetic acid, hydrogen fluoride, an aqueous hydrogen fluoride solution, triethylamine trihydrofluoride, and hydrogen fluoride pyridine salt, more preferably, the silicon-based removal reagent is an aqueous hydrogen fluoride solution. be.

[0303] In another preferred example, the inert solvent is selected from tetrahydrofuran, 2-methyltetrahydrofuran, and toluene, and more preferably, the inert solvent is tetrahydrofuran.

[0304] Another aspect of the present invention provides a method for preparing a molecule of formula 7:

[0305] [ka]

[0306] The method includes the steps of: In an inert solvent, a compound represented by the following formula 7i is subjected to a condensation reaction with a compound represented by the following formula 1d to produce a compound represented by the following formula 7:

[0307] [ka]

[0308] In another preferred example, the inert solvent is selected from the group consisting of dichloromethane, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, HMPA, or a combination thereof, preferably N,N-dimethylformamide.

[0309] In another preferred example, the condensing agent is selected from the group consisting of HATU, HBTU, TBTU, EDCI, HOAt, HOBt, CDI, TCFH, TFFH, DCC, DIC, BOP, AOP, PyAOP, BrOP, PyClOP, PyBrOP, DMTMM, or a combination thereof, and is preferably EDCI.

[0310] In another preferred example, the base is selected from the group consisting of TEA, DIPEA, NMM, DBU, DMAP, pyridine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, imidazole, N-methylimidazole, or a combination thereof, preferably 2,4,6-trimethylpyridine.

[0311] Another aspect of the present invention provides a method for preparing a molecule of formula 7i:

[0312] [ka]

[0313] The method includes the steps of: In an inert solvent, a compound of formula 14j is reacted with a compound of formula 8e to obtain a compound of formula 7i.

[0314] [ka]

[0315] In another preferred example, the inert solvent is selected from the group consisting of acetonitrile, water, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or a combination thereof, and more preferably, tetrahydrofuran.

[0316] In another preferred embodiment, the reaction is carried out with or without a reaction base, preferably the base is selected from triethylamine, N,N-diisopropylethylamine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, DBU, NMM, more preferably the reaction does not require the participation of a base; In another aspect of the present invention, the present invention further provides a method for preparing a molecule of formula 14j:

[0317] [ka]

[0318] The method includes the steps of:

[0319] [ka]

[0320] (1) In an inert solvent, a compound represented by the following formula 7g is subjected to a condensation reaction with a compound represented by the following formula 14h to obtain a compound represented by the following formula 14i;

[0321] [ka]

[0322] In another preferred example, the inert solvent is selected from the group consisting of acetonitrile, water, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, ethyl acetate, or a combination thereof, more preferably ethyl acetate.

[0323] In another preferred embodiment, the reaction is carried out with or without a reaction base, preferably the base is selected from triethylamine, N,N-diisopropylethylamine, 2,6-dimethylpyridine, 2,4,6-trimethylpyridine, DBU, NMM, more preferably the reaction does not require the participation of a base; (2) Removal of t-butyl protection from a compound of formula 14i below under acidic conditions in an inert solvent to give a compound of formula 8g;

[0324] [ka]

[0325] In another preferred example, the inert solvent is selected from the group consisting of dichloromethane, tetrahydrofuran, dioxane, ethyl acetate, toluene, or a combination thereof, more preferably toluene.

[0326] In another preferred example, the acid is selected from the group consisting of hydrochloric acid, sulfuric acid, hydrogen chloride ethyl acetate solution, hydrogen chloride tetrahydrofuran solution, hydrogen chloride dioxane solution, trifluoroacetic acid, trifluoromethanesulfonic acid, methanesulfonic acid, and dichloroacetic acid, and preferably, the acid is selected from trifluoroacetic acid; (3) In an inert solvent, a compound represented by the following formula 8g is subjected to a condensation reaction with pentafluorophenol in the presence of a condensing agent to obtain a compound represented by formula 14j:

[0327] [ka]

[0328] In another preferred example, the condensing agent is selected from the group consisting of HATU, HBTU, TBTU, EDCI, HOAt, HOBt, CDI, TCFH, TFFH, DCC, DIC, BOP, AOP, PyAOP, BrOP, PyClOP, PyBrOP, DMTMM, or a combination thereof, and is preferably EDCI.

[0329] The inert solvent is selected from the group consisting of dichloromethane, tetrahydrofuran, 2-methyltetrahydrofuran, toluene, or a combination thereof, more preferably dichloromethane.

[0330] Another aspect of the present invention provides a compound represented by formula 14b:

[0331] [ka]

[0332] Another aspect of the present invention provides a compound represented by formula 14c:

[0333] [ka]

[0334] Another aspect of the present invention provides a compound represented by formula 14g:

[0335] [ka]

[0336] Another aspect of the present invention provides a compound represented by formula 14i:

[0337] [ka]

[0338] Another aspect of the present invention provides a compound represented by formula 14j:

[0339] [ka]

[0340] Another aspect of the present invention provides a compound represented by formula 14k:

[0341] [ka]

[0342] Another aspect of the present invention provides a compound represented by formula 15c:

[0343] [ka]

[0344] Another aspect of the present invention provides a compound represented by formula 15d:

[0345] [ka]

[0346] Another aspect of the present invention provides a compound represented by formula 15f:

[0347] [ka]

[0348] Another aspect of the present invention provides a compound represented by formula 15g:

[0349] [ka]

[0350] Another aspect of the present invention provides a compound represented by formula 7c:

[0351] [ka]

[0352] Another aspect of the present invention provides a compound represented by formula 7e:

[0353] [ka]

[0354] Another aspect of the present invention provides a compound represented by formula 7h:

[0355] [ka]

[0356] Another aspect of the present invention provides a compound represented by formula 8b:

[0357] [ka]

[0358] Another aspect of the present invention provides a compound represented by formula 8e:

[0359] [ka]

[0360] Another aspect of the present invention provides a compound represented by formula 8g:

[0361] [ka]

[0362] Another aspect of the present invention provides a compound represented by formula 9b:

[0363] [ka]

[0364] Another aspect of the present invention provides a compound represented by formula 9c:

[0365] [ka]

[0366] Another aspect of the present invention provides a compound represented by formula 9e:

[0367] [ka]

[0368] Another aspect of the present invention provides a compound represented by formula 9f:

[0369] [ka]

[0370] Another aspect of the present invention provides a compound represented by formula 9g:

[0371] [ka]

[0372] Another aspect of the present invention provides a compound represented by formula 9h:

[0373] [ka]

[0374] Another aspect of the present invention provides compounds shown below in formula 9i:

[0375] [ka]

[0376] [Effects of the invention] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (e.g., in the Examples) can be combined with each other to form new or preferred technical solutions, which will not be repeated here due to space limitations. DETAILED DESCRIPTION OF THE INVENTION

[0377] Based on extensive research over a long period of time, the present inventors have prepared a class of ligand-coupled drugs with novel structures, drug molecules (toxins) suitable for ligand conjugation, connector precursors suitable for ligand conjugation, and ligand conjugate precursors suitable for ligand conjugation, which have good stability, excellent therapeutic effects, and high safety. Based on the above findings, the present inventors have completed the present invention.

[0378] term As used herein, the term "alkyl group" includes straight-chain or branched-chain alkyl groups. For example, a C1-C8 alkyl group refers to a straight-chain or branched-chain alkyl group having 1 to 8 carbon atoms, such as a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t-butyl group, etc.

[0379] As used herein, the term "alkenyl group" includes straight-chain and branched-chain alkenyl groups. For example, a C2-C6 alkenyl group refers to a straight-chain or branched-chain alkenyl group having from 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.

[0380] As used herein, the term "alkynyl group" includes straight-chain and branched-chain alkynyl groups. For example, a C2-C6 alkynyl group refers to a straight-chain or branched-chain alkynyl group having from 2 to 6 carbon atoms, such as an ethynyl group, a propynyl group, a butynyl group, or the like.

[0381] As used herein, "C3-C 10 The term "cycloalkyl group" refers to a cycloalkyl group having from 3 to 10 carbon atoms. It may be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. It may be bicyclic, such as bridged or spirocyclic.

[0382] As used herein, the term "C1-C8 alkylamino group" refers to an amino group substituted with a C1-C8 alkyl group, which may be mono- or di-substituted, such as methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, isobutylamino group, t-butylamino group, dimethylamino group, diethylamino group, dipropylamino group, diisopropylamino group, dibutylamino group, diisobutylamino group, di-t-butylamino group, and the like.

[0383] As used herein, the term "C1-C8 alkoxy group" refers to a group consisting of 1 to 8 alkyl groups. It refers to a straight or branched chain alkoxy group having carbon atoms, such as a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a t-butoxy group, and the like.

[0384] As used herein, the term "3- to 10-membered heterocycloalkyl group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O" refers to a saturated or partially saturated cyclic group having 3 to 10 atoms, of which 1 to 3 heteroatoms are selected from the group consisting of N, S, and O. It may be a monocyclic or bicyclic group in the form of a bridged ring or a spirocyclic ring. Specific examples include oxetane, azetidine, tetrahydro-2H-pyranyl group, piperidinyl group, tetrahydrofuranyl group, morpholinyl group, and pyrrolidinyl group.

[0385] As used herein, "C6-C 10 The term "aryl group" refers to an aryl group having 6 to 10 carbon atoms, such as a phenyl group or a naphthyl group or similar group.

[0386] As used herein, the term "5- to 10-membered heteroaryl group having 1 to 3 heteroatoms selected from the group consisting of N, S, and O" refers to a cyclic aromatic group having 5 to 10 atoms, 1 to 3 of which are heteroatoms selected from the group consisting of N, S, and O. It may be a monocyclic or fused ring form. Specific examples include pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, pyrrolyl, pyrazolyl, imidazolyl, (1,2,3)-triazolyl, (1,2,4)-triazolyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, etc.

[0387] Unless otherwise specified, in this specification, a chain group (e.g., an alkyl group, a halogenated alkyl group, or a deuterated alkyl group) has 1 to 10 carbon atoms, and preferably 1 to 6 carbon atoms or 1 to 4 carbon atoms; in this specification, a non-aromatic cyclic group (e.g., a cycloalkyl group, a heterocycloalkyl group, an alicyclic group, an aliheterocyclic group, etc.) has 3 to 12 members, preferably 3 to 8 members, or 3 to 6 members; and in this specification, an aromatic cyclic group (e.g., an aryl group, a heteroaryl group, etc.) has 5 to 15 members, such as a 6 to 10-membered aryl group, a 5 to 7-membered heteroaryl group, or a 5 to 10-membered heteroaryl group.

[0388] Unless otherwise specified, the groups described in the present invention are "substituted or unsubstituted", and all groups in the present invention are substituted or unsubstituted, including halogen, nitrile, nitro, hydroxyl, amino, C1-C6 alkyl-amino, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, halogenated C2-C6 alkenyl, halogenated C2-C6 alkynyl, halogenated C1-C6 alkoxy, allyl, benzyl, C6-C 12 It may be substituted by a substituent selected from the group consisting of an aryl group, a C1-C6 alkoxy-C1-C6 alkyl group, a C1-C6 alkoxy-carbonyl group, a phenoxycarbonyl group, a C2-C6 alkynyl-carbonyl group, a C2-C6 alkenyl-carbonyl group, a C3-C6 cycloalkyl-carbonyl group, a C1-C6 alkyl-sulfonyl group, and the like.

[0389] As used herein, "halogen" or "halogen atom" refers to F, Cl, Br, and I. More preferably, the halogen or halogen atom is selected from F, Cl, and Br. "Halogenated" refers to substitution with an atom selected from F, Cl, Br, and I.

[0390] Unless otherwise specified, the structural formulae depicted in this invention include all isomeric forms (e.g., enantiomers, diastereomeric forms), such as R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, etc. The present invention is intended to include stereoisomers and geometric isomers (or conformational isomers) of the compounds of the present invention. Accordingly, all individual stereochemical isomers or enantiomeric, diastereomeric or geometric isomeric (or conformational isomer) mixtures of the compounds of the present invention are within the scope of the present invention.

[0391] As used herein, the term "tautomer" refers to structural isomers of different energies that can be interconverted across a low energy barrier. For example, proton tautomers (i.e., proton migration) include interconversions via migration of a proton, such as 1H-indazole and 2H-indazole. Valence tautomers include interconversions via recombination of some bond electrons.

[0392] As used herein, the term "hydrate" refers to a complex formed when a compound of the present invention is coordinated with water. The compounds of the present application can be prepared by various synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining specific embodiments with other chemical synthetic methods, and equivalent substitution methods known to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present application.

[0393] The solvents used in this application are commercially available and the compounds are named artificially or by ChemDraw® software; for commercially available compounds, the supplier's catalog name is used.

[0394] In this application, the term "ligand" generally refers to a large molecule compound that can recognize and bind to an antigen or receptor associated with a target cell. The role of the ligand is to present a drug to a target cell population that binds to the ligand. These ligands include, but are not limited to, protein hormones, lectins, growth factors, antibodies, or other molecules that can bind to cells, receptors, and / or antigens. In this application, the ligand is referred to as Ab, and the ligand-antigen is attached to the unit via a heteroatom on the ligand to form a linking bond. The ligand may be an antibody or an antigen-binding fragment thereof. The antibody may be selected from a chimeric antibody, a humanized antibody, a fully human antibody, or a murine antibody, and may be a monoclonal antibody. For example, the antibody may be an antibody or an antigen-binding fragment thereof that targets a target point selected from the group consisting of HER2, TROP2, PSMA, FR-α, B7H3, and IGF-1R.

[0395] In this application, the terms "Trop2" and "TROP2" generally refer to a single-pass, type I plasma membrane protein. In this application, the term "Trop2" can further include homologs, variants, and isoforms of Trop2, including splice isoforms. The term "Trop" further includes proteins having one or more sequences of Trop2 homologs, variants, and isoforms, and fragments of such sequences, so long as the variant proteins (including isoforms) are such variant proteins. Trop2 can be human Trop2. For example, Uniprot Accession No. P09758 provides a description of Trop2 and its sequence.

[0396] In this application, the term "HER2" generally refers to human epidermal growth factor receptor 2 (HER2). For example, the term "HER2" refers to any native HER2 from any human. The term further includes "full-length" and unprocessed HER2, as well as any form of HER2 processed within a cell (e.g., the mature protein). The term further includes naturally occurring variants and isoforms of HER2, such as splice variants or allelic variants. For example, Uniprot Accession No. P04626 provides a description of HER2 and its sequence.

[0397] As used herein, the term "PSMA" refers to glutamate carboxypeptidase II. As used herein, the term "PSMA" can further include homologs, variants, and isoforms of PSMA. The term "PSMA" also includes proteins having one or more sequences of PSMA homologs, variants, and isoforms of various different origins (e.g., human origin), and fragments of such sequences. For example, Uniprot Accession No. Q04609 provides a description of PSMA and its sequences.

[0398] In the present application, the term "FR-α" refers to folate receptor α encoded by FOLR1. In the present application, the term "FR-α" can further include homologs, variants, and isoforms of FR-α. The term "FR-α" further includes proteins having one or more sequences of FR-α homologs, variants, and isoforms of various different origins (e.g., human origin), and fragments of said sequences. For example, Uniprot Accession No. P15328 provides a description of FR-α and the sequence.

[0399] In this application, the term "B7H3" refers to CD276. In this application, the term "B7H3" can include any homologs, variants, and isoforms thereof, including all possible forms of its expression in humans (e.g., 2Ig and 4Ig). The term "B7H3" further includes proteins having one or more sequences of B7H3 homologs, variants, and isoforms of various origins (e.g., human origin), and fragments of such sequences. For example, Uniprot Accession No. Q5ZPR3 provides a description of B7H3 (CD276) and its sequence.

[0400] In this application, the term "IGF-1R" refers to insulin-like growth factor I receptor. In this application, the term "IGF-1R" can further include its homologs, variants, and isoforms. The term "IGF-1R" further includes proteins having one or more sequences of IGF-1R homologs, variants, and isoforms of various different origins (e.g., human origin), and fragments of said sequences. For example, Uniprot Accession No. P08069 provides a description of IGF-1R and sequences.

[0401] In the present application, the term "peptide residue" generally refers to a residue formed by the combination of one or more amino acid residues. For example, one or more amino acids of a polypeptide residue can be optionally substituted. For example, a polypeptide residue of the present application can include glycine-glycine-phenylalanine-glycine (Gly-Gly-Phe-Gly).

[0402] In this application, the term "drug unit" generally refers to a chemical moiety that binds directly or indirectly to an antibody or antigen-binding fragment to form an immunoadduct. For example, a "drug unit" includes, but is not limited to, a compound having anti-tumor activity as described herein. For example, a drug unit includes a topoisomerase inhibitor.

[0403] In this application, the term "compound with anti-tumor activity" generally refers to a compound that has the ability to reduce the proliferation rate, survival, or metastatic activity of tumor cells. For example, anti-tumor activity is demonstrated by a decrease in the growth rate of abnormal cells or a stabilization or reduction in tumor size during treatment, or by an increase in survival time with treatment compared to an untreated control. Anti-tumor activity can be assessed using recognized in vitro or in vivo tumor models, such as xenograft models.

[0404] In this application, the term "comprising" generally refers to the inclusion of the specifically stated feature but not the exclusion of other elements. The terms "equivalent to" and "equivalent to" generally refer to the inclusion of the numerical value itself.

[0405] In this application, the term "about" generally refers to a variation within 0.5% to 10% above or below a specified value, for example, a variation within 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below a specified value.

[0406] In this application, the compounds of the present application include their tautomers, meso-isomers, racemates, enantiomers, and / or diastereomers.

[0407] In this application, certain atoms of the compounds of this application can be present in one or more isotopic forms. For example, hydrogen can be present as hydrogen ( 1 H), deuterium ( 2 H) and tritium ( 3 H), and carbon exists in three different isotopes ( 12 C. 13 C and 14 Examples of isotopes that can be incorporated into compounds of the present application include: 15 N, 18 O. 17 O. 18 F, 32 P,33 P, 129 I, 131 I, 123 I, 124 I, 125 I, or similar isotopes. Thus, the compounds of the present application may be in enriched form with one or more of these isotopes compared to the natural abundance of these isotopes. As is well known to those skilled in the art, such isotopically enriched compounds can be used in a variety of applications. For example, deuterium ( 2 Substitution with heavier isotopes such as 3H may offer certain therapeutic advantages due to increased metabolic stability.

[0408] In the present application, the term "pharmaceutical composition" generally refers to a mixture of one or more compounds described in the present application or physiologically / pharmaceutically acceptable salts or prodrugs thereof with other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. Pharmaceutical compositions can facilitate administration to an organism and favor the absorption of active ingredients, thereby enabling them to exert their biological activity. Conventional pharmaceutical composition preparations can refer to common techniques in the art.

[0409] In the present application, the term "pharmaceutically acceptable salt" or "pharmaceutically acceptable salt" generally refers to a salt of a compound or ligand-drug conjugate of the present application, or a salt of a compound described in the present application. Such salts may be safe and / or effective when used in mammals and may have desirable biological activity. The antibody-antibody drug coupling compound of the present application may form a salt with an acid. Non-limiting examples of pharmaceutically acceptable salts include hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfite, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, hydrogen citrate, tartrate, maleate, fumarate, formate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, or p-toluenesulfonate.

[0410] As used herein, the term "conjugate" generally refers to compounds prepared by one or more chemical reactions of the compounds of the present application, or compounds linked together via one or more linking structures, such as a bridge, spacer, or linking moiety.

[0411] In this application, the term "pharmaceutically acceptable carrier" generally refers to a carrier for administering therapeutic agents, such as antibodies or polypeptides, genes, and other therapeutic agents. The term refers to any drug carrier that can be administered without inducing the production of antibodies harmful to the individual receiving the composition and without undue toxicity. For example, a pharmaceutically acceptable carrier is different from a nucleic acid carrier used to contain a target gene in genetic engineering. Suitable carriers can be large, slowly metabolized molecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, lipid aggregates, and inactivated virus particles. These carriers are well known to those skilled in the art. Pharmaceutically acceptable carriers in therapeutic compositions include liquids such as water, saline, glycerol, and ethanol. These carriers can also contain auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, and the like.

[0412] In this application, the term "antibody" generally includes monoclonal antibodies, polyclonal antibodies, dimers, multimers, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity. Antibodies can be murine, human, humanized, chimeric, or derived from other species. Antibodies are proteins produced by the immune system that can recognize and bind to specific antigens. A target antigen typically has multiple binding sites, also called epitopes, distinguished by the CDRs of various antibodies. Each antibody that specifically binds to a different epitope has a different structure. Thus, an antigen can have one or more corresponding antibodies. Antibodies include full-length immunoglobulin molecules or immunologically active portions of full-length immunoglobulin molecules, i.e., molecules containing an antigen or portion thereof that specifically binds to a target of interest, including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with autoimmune diseases. The immunoglobulins described herein can have any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2, or any mutation thereof), or subclass of immunoglobulin molecule. Immunoglobulins can be obtained from any species. However, in one embodiment, the immunoglobulin is of human, murine, or rabbit origin. An "antibody fragment" comprises a portion of a full-length antibody, generally the antigen-binding or variable region thereof. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; minibodies; fragments prepared by an Fab expression library; anti-idiotypic (anti-Id) antibodies; CDRs (complementarity-determining regions); and any epitope-binding fragments of the above that immunospecifically bind to a cancer cell antigen, a viral antigen, or a microbial antigen; single-chain antibody molecules; and multispecific antibodies formed by antibody fragments. The antibody constituting the antibody-drug conjugate in the present application can maintain its original antigen-binding ability in the wild state, and therefore, the antibody in the present application can specifically bind to, for example, an antigen.The antigens involved include, for example, tumor-associated antigens (TAAs), cell surface receptor proteins and other cell surface molecules, cell survival regulators, cell proliferation regulators, molecules associated with tissue growth and differentiation (e.g., with known or predicted functionality), lymphokines, cytokines, molecules involved in regulating cell circulation, molecules involved in angiogenesis, and molecules associated with angiogenesis. For example, known antigens bound by antibodies may be one or a subset of the above categories, while other subsets include molecules / antigens with other unique properties (compared to the target antigen). Antibodies used in antibody-drug conjugates include, but are not limited to, antibodies against cell surface receptors and tumor-associated antigens. Such tumor-associated antigens are well known in the art and can be prepared using antibody preparation methods and information well known in the art. These targets may be specifically expressed on the surface of one or more cancer cells, while being expressed poorly or not at all on the surface of one or more non-cancerous cells. Typically, such tumor-associated polypeptides may be overexpressed on the surface of cancer cells relative to the surface of non-cancerous cells.

[0413] In this application, the term "chimeric antibody" generally refers to an antibody formed by fusing the variable region of a mouse-derived antibody with the constant region of a human antibody, which can reduce the immune response elicited by mouse-derived antibodies. To establish a chimeric antibody, a hybridoma secreting a specific mouse-derived monoclonal antibody can be established, and then the variable region genes can be cloned from the mouse hybridoma cells, and if necessary, the constant region genes of a human antibody (the antibody can be a human antibody). The mouse variable region genes and human constant region genes can be linked to form chimeric genes, which can then be inserted into an expression vehicle and the chimeric antibody molecule can be expressed in eukaryotic or prokaryotic systems.

[0414] In this application, the term "humanized antibody," also referred to as a CDR-grafted antibody, generally refers to an antibody generated by grafting mouse CDR sequences onto a human antibody variable region framework, i.e., a different type of human germline antibody framework sequence. Because they retain a large amount of mouse protein components, they can overcome the heterologous reactions caused by chimeric antibodies. Such framework sequences can be obtained from public DNA databases or published references that contain germline antibody gene sequences. For example, germline DNA sequences of human heavy and light chain variable region genes can be found in the "VBase" human germline sequence database.

[0415] In this application, the term "fully human antibody," also referred to as "fully human monoclonal antibody," refers to an antibody in which both the variable and constant regions are human, eliminating immunogenicity and toxic side effects. The development of monoclonal antibodies goes through four stages: mouse-derived monoclonal antibodies, chimeric monoclonal antibodies, humanized monoclonal antibodies, and fully human monoclonal antibodies. The antibodies or ligands described in this application may be fully human monoclonal antibodies. Related technologies for preparing fully human antibodies include human hybridoma technology, EBV-transformed B lymphocyte technology, phage display technology, transgenic mouse antibody preparation technology, and single B cell antibody preparation technology.

[0416] In this application, the term "CDR" refers to one of the six hypervariable regions in the variable domain of an antibody that primarily mediates antigen binding. One of the most commonly used definitions of the six CDRs is provided by Kabat EA et al., or Chothia et al. and MacCallum et al. As used in this application, the Kabat definition of CDR can be applied to CDR1, CDR2, and CDR3 of the light chain variable domain (LCDR1, LCDR2, LCDR3 or L1, L2, L3) and CDR1, CDR2, and CDR3 of the heavy chain variable domain (HCDR1, HCDR2, HCDR3 or H1, H2, H3).

[0417] In the present application, the term "a group capable of coupling to a mercapto group" generally refers to compound A having a mercapto group, compound B having a group capable of coupling to a mercapto group, and compound B reacting with the mercapto group of compound A via the group capable of coupling to a mercapto group, thereby realizing bonding between compound A and compound B.

[0418] In the present application, the term "connector" generally refers to a chemical fragment or bond having one end bound to one group and the other end bound to another group, which can be linked to a drug and / or ligand after being linked to another connector. The directly or indirectly linked ligand means that the group can be directly linked to the ligand via a covalent bond or can be linked to the ligand via a connector. For example, the connector can be a structure shown in the connectors described in the present application. For example, a chemical fragment or bond containing an acid-labile linker structure (e.g., hydrazone), a protease-sensitive (e.g., peptidase-sensitive) linker structure, a photolabile linker structure, a dimethyl linker structure, or a disulfide-containing linker structure can be used as a connector.

[0419] In this application, the term "linking group" generally refers to a group that has the ability to link to another group. For example, a compound having a linking group can be linked to another group by a coupling reaction between the linking group and another group. For example, a maleimide group can function as a linking group.

[0420] In the present application, the term "disease associated with the expression" of a particular target point generally refers to the occurrence and / or progression of the disease being related to the expression level of the target point. For example, the expression level of the particular target point in cells of a diseased region, such as a particular tissue or organ of a patient, is increased, i.e., highly expressed, compared to the expression level in normal cells of the tissue or organ. Or, for example, the expression level of the particular target point in cells of a diseased region, such as a particular tissue or organ of a patient, is decreased, i.e., underexpressed, compared to the expression level in normal cells of the tissue or organ. Or, for example, cells of a diseased region, such as a particular tissue or organ of a patient, express the particular target point, i.e., are positive. Or, for example, cells of a diseased region, such as a particular tissue or organ of a patient, do not express the particular target point, i.e., are negative. For example, target point expression characteristics can be determined by standard measurements known in the art.

[0421] In this application, the term "effective amount" generally refers to the amount of a therapeutic agent to treat, alleviate, or prevent a targeted disease or condition, or an amount that exhibits a detectable therapeutic or preventative effect. The precise effective amount for a particular subject will vary depending on the subject's size and health, the nature and severity of the condition, and the therapeutic agent and / or combination of therapeutic agents selected for administration. Thus, it is not useful to pre-specify an exact effective amount. However, for a particular situation, the effective amount can be determined using routine experimentation and is within the judgment of the clinician.

[0422] Unless otherwise specified, all compounds presented in this application are intended to include all possible optical isomers, such as a single chiral compound or a mixture of various different chiral compounds (i.e., a racemate). In all compounds in this application, each chiral carbon atom can optionally be in the R or S configuration, or a mixture of the R and S configurations.

[0423] In this application, the term "compound of the application" generally refers to the compound of the application. The term further includes various crystalline forms, pharmaceutically acceptable salts, hydrates, or solvates of the compound of the application.

[0424] When trade names are used herein, the trade names are intended to include formulations of the trade name product, its corresponding generic drug products, and the active ingredients of the trade name product.

[0425] Pharmaceutical compositions and methods of administration Since the compound of the present invention has excellent inhibitory activity against tumor cell proliferation, the compound of the present invention and its various crystalline forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, as well as pharmaceutical compositions containing the compound of the present invention as a main active ingredient, can be used to prevent and / or treat (stabilize, alleviate or cure) diseases associated with tumor cell proliferation.

[0426] The pharmaceutical composition of the present invention contains a compound of the present invention and a pharmaceutically acceptable excipient or vector within a safe and effective amount. Here, "safe and effective amount" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1 to 2000 mg of the compound / agent of the present invention, more preferably 1 to 200 mg of the compound / agent of the present invention. Preferably, the "single agent" is a single capsule or tablet, or a unit-dose injection preparation.

[0427] A "pharmaceutically acceptable vector" is a vector that is suitable for human use, of sufficient purity and sufficient quality. "Compatibility" refers to one or more compatible solid or liquid fillers or gel substances that must have low toxicity. "Compatibility" refers to the ability of the components of the composition to blend with each other without significantly reducing the efficacy of the compounds of the present invention. Some examples of pharmaceutically acceptable vectors include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween®), wetting agents (e.g., sodium dodecyl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0428] The administration route of the compound or pharmaceutical composition of the present invention is not particularly limited, and typical administration routes include (but are not limited to) oral and parenteral (intravenous, intramuscular, or subcutaneous).

[0429] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or vector), such as sodium citrate or dicalcium phosphate, or with (a) a filler or compatibilizer, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) a binder, such as hydroxymethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) a humectant, such as glycerin; (d) agar, calcium carbonate, potato starch, or the like. The formulation may be mixed with ingredients such as disintegrating agents such as potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, retarders such as paraffin, absorption accelerators such as quaternary amine compounds, wetting agents such as cetyl alcohol and glyceryl monostearate, adsorbents such as kaolin, and lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, and sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain a buffering agent.

[0430] Solid dosage forms such as tablets, sugar pills, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials known in the art. They can contain opacifying agents, and the release of the active compound or compounds of such compositions can be delayed in a specific part of the digestive tract. Examples of embedding materials that can be used include polymeric substances and waxes. If necessary, the active compound can be formed into microcapsules with one or more of the above-mentioned excipients.

[0431] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms can contain inert diluents conventionally used in the art, such as water or other solvents, and solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0432] In addition to these inert diluents, compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and perfuming agents. In addition to the active compound, the suspension may contain, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, Suspending agents such as aluminum methoxide and agar or mixtures of these substances may be included.

[0433] Compositions for parenteral injection can include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous vehicles, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0434] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents. When administered in combination, the pharmaceutical composition further comprises one or more (two, three, four, or more) other pharmaceutically acceptable therapeutic agents, which may be used simultaneously, separately, or sequentially with the compounds of the present invention to prevent and / or treat diseases associated with tumor cell proliferation.

[0435] When a pharmaceutical composition is used, a safe and prevalent amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, and the dosage at the time of administration is the considered effective dose, and for a person weighing 60 kg, the daily dose is usually 1 to 2000 mg, preferably 1 to 500 mg. Of course, the specific dosage must also take into account factors such as the route of administration and the patient's health condition, all of which are within the skill of a skilled physician.

[0436] Preparation process of toxin molecule intermediates The present invention further provides a novel preparation process for synthetic intermediate 14d of toxin molecule 1d (compound 1k in WO2022262789A1), the specific preparation process of which is as described above. The synthetic route disclosed in the prior art (WO2022262789A1) is as follows:

[0437] [ka]

[0438] Compared with the above-mentioned prior art, the preparation process of the present invention replaces one of the two TES silicon-based protecting groups in the prior art with a TIPS silicon-based protecting group, and has the following main advantages:

[0439] 1. The intermediates 14b and 14c in the synthetic route of the present application are more stable than the intermediates 1i and 1j in the above-mentioned prior art, and are easier to scale up production and store for long periods of time.

[0440] 2. Intermediates 14b and 14c in the synthetic route of the present application can be separated and purified, and quality control is easy. Intermediates 1i and 1j in the above prior art are crude and suitable for the next step of the reaction. It is used as a product.

[0441] 3. The yield of the synthetic route of the present application is increased from 49% to 73% compared to the yield of the above prior art (WO2022262789A1). Preparation process of toxin molecules The present invention further provides a novel preparation process for toxin molecule 1d (compound 1n-P1 in WO2022262789A1), the synthesis route in the prior art (WO2022262789A1) is as follows:

[0442] [ka]

[0443] Compared with the above prior art, the preparation process of the present invention replaces the acetyl Ac protecting group in the prior art with an Fmoc protecting group, and has the following main advantages:

[0444] In the synthetic route of the present application, the acetyl protecting group of 1l in the synthetic route of the above-mentioned prior art is replaced with an Fmoc protecting group to obtain compound 14g, thereby improving the synthetic yield of compound 1d (compound 1n-P1 in WO2022262789A1) from 7.9% to 15.6%.

[0445] The deacetylation step of compound 1m in the synthetic route of the above-mentioned prior art (WO2022262789A1) has harsh reaction conditions, poor reproducibility, and incomplete removal of the extended protecting group, whereas the reaction conditions for the Fmoc deprotection step in method 14k of the present application are milder and more suitable for large-scale production.

[0446] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and do not limit the scope of the present invention. In the following examples, experimental methods without specific conditions are usually performed according to conventional conditions or conditions suggested by manufacturers. Unless otherwise specified, percentages and parts are calculated by weight.

[0447] Abbreviations

[0448] [Table 1] TIFF2025538619000227.tif224170TIFF2025538619000228.tif223170TIFF2025538619000229.tif222170TIFF20255386190 00230.tif221170TIFF2025538619000231.tif223170TIFF2025538619000232.tif228170TIFF2025538619000233.tif228170

[0449] Example Example 1.1

[0450] [ka]

[0451] Phase 1 Compound 1b (342 mg, 0.81 mmol) and 1a (500 mg, 0.85 mmol) were dissolved in a mixed solvent of acetonitrile (3.42 mL) and water (6.84 mL). The mixture was cooled to 0°C, and N,N-diisopropylethylamine (83 mg, 0.64 mmol) was added dropwise with stirring. After the addition was completed, the mixture was allowed to react for 3 hours with continuous stirring. The reaction mixture was directly separated and purified by preparative chromatography to give 1c (283 mg) in a yield of 39%.

[0452] MS-ESI calculated value [M+Na] + =1020, actual measured value 1020. Phase 2 Compound 1c (85 mg, 0.19 mmol) was dissolved in N,N-dimethylformamide (1.70 mL), trifluoroacetic acid (22 mg, 0.19 mmol) was added, and the mixture was cooled to 0°C. 1d (226 mg, 0.23 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (72 mg, 0.38 mmol), and 2,4,6-trimethylpyridine (23 mg, 0.19 mmol) were added sequentially. After the addition was complete, the reaction system was maintained at 0°C to 10°C and allowed to react for 1 hour with continuous stirring. The reaction solution was then diluted with HCl.aq (0.05M) and adjusted to pH 6~7, the liquid phase was separated and purified, and compound 1 (134 mg) was obtained. Yield: 49%.

[0453] 1 H NMR(400MHz,DMSO-d6)δ 8.66(t,J=6.4Hz,1H),8.60(d,J=8.8Hz,1H),8.28(t,J=6.0Hz,1H),8.15(t,J=5.6Hz,1H),8.09(d,J=8.0Hz,1H),8.04-7.95(m,2H ),7.84-7.77(m,2H),7.28-7.14(m,5H),6.99(s,2H),6.69(s,1H),5.91(d,J=16.8Hz,1H),5.70-5.62(m,1H),5.56-5.45(m,2H),5 .35(d,J=18.8Hz,1H),4.68(d,J=6.8Hz,2H),4.51-4.42(m,1H),4.17-4.04(m,2H),3.80-3.55(m,12H),3.55-3.42(m,32H),3.17- 3.10(m,2H),3.06-2.98(m,1H),2.80-2.71(m,1H),2.41-2.28(m,7H),2.27-2.15(m,2H),1.94-1.82(m,2H),0.86(t,J=7.2Hz,3H).

[0454] Example 1.2

[0455]

change

[0456] Stage 1 2a (14.72 g, 150.16 mmol) was dissolved in acetone (118 mL), and 2b (20.00 g, 150.16 mmol) was added under ice bath conditions. The mixture was stirred for 5 min, and then monitored for completion by TLC. The reaction mixture was concentrated to give a solid crude product. The crude product was dissolved in acetic anhydride (28 mL), and sodium acetate (24.63 g, 300.32 mmol) was added. The reaction mixture was heated to 90 °C and refluxed for 2 h. The reaction mixture was filtered to remove insoluble solids. The filter cake was eluted with toluene, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was isolated by silica gel column chromatography (EA: hexane = 0-100%) to give 2c (17.43 g) in 43% yield.

[0457] MS-ESI calculated value [M+Na] + =236, actual value 236. Phase 2 2c (12.95 g, 60.74 mmol) and 2d (9.00 g, 60.74 mmol) were dissolved in toluene (180 mL), p-toluenesulfonic acid (2.10 g, 12.15 mmol) was added, and the mixture was heated to 90 °C and refluxed for 2 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give a residue. The residue was dissolved in ethyl acetate, and the organic phase was washed successively with saturated sodium bicarbonate and saturated brine (200 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a crude product. The crude product was separated by silica gel column chromatography (EA: hexane = 0 to 100%) to give 2e ( 5.53 g) was obtained, yield: 33%.

[0458] MS-ESI calculated value [M+H] + =270, actual value 270. Phase 3 2e (3.87 g, 14.37 mmol) was dissolved in THF (77.4 mL), and lithium hydroxide (1.37 g, 57.49 mmol) was dissolved in HO (38.7 mL) and added to the above solution. The mixture was stirred at room temperature for 30 minutes. Ethyl acetate (15 mL) was added to the reaction solution, and HCl aqThe pH was adjusted to about 2 with 1N aqueous sodium chloride, the aqueous phase was extracted with ethyl acetate (38 mL × 3), the organic phases were combined, the organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 2f (3.06 g).

[0459] MS-ESI calculated value [M+H] + =260, actual value 260. Phase 4 N-Hydroxysuccinimide (6.49 g, 56.37 mmol) was dissolved in N,N-dimethylformamide (36.5 mL), trifluoroacetic anhydride (11.84 g, 56.37 mmol) was added dropwise in an ice bath, and after stirring for 30 minutes, 2,4,6-trimethylpyridine (6.83 g, 56.37 mmol) was added dropwise. After the addition was complete, the mixture was stirred for 40 minutes, and the reaction mixture was recorded as A and stored. Crude product 2f (3.06 g) was dissolved in N,N-dimethylformamide (36.5 mL), 2,4,6-trimethylpyridine (3.41 g, 28.15 mmol) was added dropwise in an ice bath, and after stirring for 30 minutes in an ice bath, reaction mixture A was added dropwise. The reaction mixture was allowed to warm to room temperature and reacted for 24 hours with stirring. Dichloromethane (180 mL) was added to the reaction mixture, and HCl was added. aq After adding 140 mL of 0.7N hexane, the mixture was stirred for 30 minutes, and the layers were separated. The aqueous phase was extracted with 70 mL of dichloromethane. The combined organic phases were adjusted to pH 5-7 with water, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (EA: hexane = 0-100%) to obtain 2g (3.54g), yield: 78%.

[0460] MS-ESI calculated value [M+H] + =339, actual value 339. 1H NMR(400MHz,CDCl3)δ 6.77(s,2H),4.79(t,J=4.8Hz,1H),4.49-4.40(m,2H),3.91(t,J=11.6Hz,2H),3.79-3.70(m,2H),3.45-3.34(m,1H),2.86(s,4H).

[0461] Stage 5 2h (850 mg, 3.38 mmol) and 2g (1.20 g, 3.55 mmol) were dissolved in N,N-dimethylformamide (8.5 mL), and N,N-diisopropylethylamine (437 mg, 3.38 mmol) was added dropwise in an ice bath. The mixture was stirred for 2 hours while controlling the temperature between 0 and 10°C. The reaction mixture was separated and purified by liquid phase separation to give 2i (740 mg) in a 46% yield.

[0462] MS-ESI calculated value [M+H] + 475, actual value 475. Stage 6 2j (10.00 g, 13.59 mmol) was dissolved in THF (450 mL), wet Pd / C (2 g, 20% w / w) was added, and the mixture was stirred under a hydrogen atmosphere for 66 h. The reaction mixture was filtered, and the filter cake was eluted with a DCM / MeOH mixture. The filtrate was dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product. The crude product was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 2k (6.10 g), yield: 69%.

[0463] MS-ESI calculated value [M+H] + =668, actual value 668. Stage 7 Under a nitrogen atmosphere, 1d (2.70 g, 5.97 mmol) and trifluoroacetic acid (680 mg, 5.97 mmol) were dissolved in N,N-dimethylformamide (54 mL) and stirred in an ice bath for 10 minutes. 2k (5.40 g, 8.36 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.30 g, 11.94 mmol), and 2,4,6-trimethylpyridine (722 mg, 5.97 mmol) were then added to the reaction mixture. The temperature was controlled at 0°C to 10°C and the mixture was allowed to react for 2 hours with stirring. HCl was added to the reaction mixture in an ice bath. aq Add 0.05N (54 mL) dropwise to precipitate a solid. Dilute the suspension with 2-methyltetrahydrofuran (100 mL) and stir until dissolved. Then, separate the aqueous phase with 2-methyltetrahydrofuran (100 mL x 2). Combine the organic phases, wash with saturated brine (50 mL x 1), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (MeOH:DCM = 0-100%) to obtain 2L (6.00 g), yield: 84%.

[0464] MS-ESI calculated value [M+H] + =1079, actual value 1079. Stage 8 Dissolve 2L (6.00 g, 5.56 mmol) in a DCM / MeOH (120 mL / 12 mL) mixture and add diethylamine (24 mL) dropwise in an ice bath. After the addition is complete, allow the mixture to warm to room temperature and react with stirring for 4 hours. The reaction mixture is concentrated directly to give a brown solid crude product. Methyl t-butyl ether is added to form a slurry, filtered, and the filter cake is eluted three times with methyl t-butyl ether. The filtrate is concentrated to give a yellow solid crude product. A portion of the crude product is separated by preparative liquid phase separation to give 2M (150 mg). The remaining crude product is separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 2M (800 mg). Combined yield: 59%.

[0465] MS-ESI calculated value [M+H] + =857, actual value 857. Stage 9 2i (74 mg, 0.16 mmol) was dissolved in N,N-dimethylformamide (1.30 mL), and the atmosphere was purged with nitrogen gas three times. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (88 mg, 0.23 mmol) was added in an ice bath, and after stirring for 10 minutes, 2m (132 mg, 0.16 mmol) was added. After stirring until dissolved, 2,4,6-trimethylpyridine (54 mg, 0.45 mmol) was added, and the temperature was controlled at 0°C to 10°C for 2 hours. The reaction solution was then diluted with HCl. aq The pH was adjusted to 6-7 with 0.05N HCl, and the mixture was separated and purified by liquid phase separation to obtain 2 (71 mg), yield: 34%.

[0466] MS-ESI calculated value [M+H] + =1313, actual value 1313. Example 1.3

[0467] [ka]

[0468] Phase 1 Under a nitrogen atmosphere, 3a (5.00 g, 26.74 mmol) was dissolved in N,N-dimethylformamide (50 mL). The mixture was cooled to 0-5°C, and NaH (1.28 g, 32.09 mmol) was added. After stirring for 10 min, t-butyl bromide acetate (6.23 g, 32.09 mmol) was added. The mixture was maintained at 0-5°C and stirred for 2 h. Water (200 mL) was added to the reaction mixture, and the aqueous phase was extracted with ethyl acetate (200 mL). The organic phase was washed with saturated brine (200 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3b (4.60 g) in a 43% yield.

[0469] MS-ESI calculated value [M+H] + =302,304, actual value 302,304. 1H NMR(400MHz, CDCl3)δ 8.60(d,J=2.0Hz,1H),7.84(dd,J=8.4,2.4Hz,1H),7.45(d,J=8.4Hz,1H),4.69(s,2H),4.10(s,2H),1.49(s,9H).

[0470] Phase 2 Under a nitrogen atmosphere, 3b (5.00 g, 16.60 mmol), 3c (3.61 g, 19.93 mmol), Pd(dba) (0.76 g, 0.83 mmol), BINAP (1.03 g, 1.66 mmol), and cesium carbonate (13.53 g, 41.52 mmol) were dissolved in anhydrous toluene (50 mL). The mixture was heated to 80 °C and stirred for 16 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was separated by silica gel column chromatography (EA: hexane = 0-100%) to give 3d (4.50 g) in a 67% yield.

[0471] MS-ESI calculated value [M+H] + =403, actual value 403. Phase 3 3d (2.81 g, 6.98 mmol) in THF (28 mL) and HCl aq The mixture was dissolved in a mixed solvent of 1N (28 mL) and the mixture was stirred at room temperature for 3 hours. Water (100 mL) was added to the reaction mixture, and the aqueous phase was washed with ethyl acetate (100 mL x 2). The organic phase was The aqueous phase was adjusted to pH 8-9 with aqueous ammonia and then extracted with ethyl acetate (100 mL × 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3e (1.43 g), yield: 86%.

[0472] MS-ESI calculated value [M+H] + =239, actual value 239. Phase 4 3e (1.83 g, 7.68 mmol) and maleic anhydride (0.75 g, 7.68 mmol) were dissolved in acetonitrile (18.3 mL) and reacted at room temperature with stirring for 2 hours. The reaction mixture was directly spin-dried to obtain 2.45 g of a white solid intermediate. 2.45 g of the white solid intermediate was added to a reaction bottle, and acetic anhydride (5 mL) and sodium acetate (2.09 g, 15.36 mmol) were added sequentially. The mixture was reacted at room temperature with stirring for 2 hours. Water (50 mL) was added to the reaction mixture, and the aqueous phase was extracted with ethyl acetate (50 mL). The organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was separated by silica gel column chromatography (EA: hexane = 0-100%) to obtain 3f (1.97 g) in an 80% yield.

[0473] MS-ESI calculated value [M+H] + =319, actual value 319. 1 H NMR(400MHz,DMSO-d6)δ 8.51(d,J=2.0Hz,1H),7.83(dd,J=8.4,2.4Hz,1H),7.6(d,J=8.4Hz,1H),7.24(s,2H),4.67(s,2H),4.15(s,2H),1.44(s,9H).

[0474] Stage 5 Dissolve 3f (0.90 g, 2.83 mmol) in dichloromethane (9 mL), add trifluoroacetic acid (1.8 mL), and react at room temperature for 5 h with stirring. The reaction mixture was directly spun down, and the dichloromethane (45 mL × 5) was evaporated. The mixture was then concentrated in an oil pump until no visible oil remained, yielding crude product 3f (0.93 g).

[0475] MS-ESI calculated value [M+H] + =263, actual value 263. Stage 6 Add 3g (0.06g), 2m (0.20g, 0.23mmol) and N,N-dimethylformamide (4mL) sequentially to the reaction bottle, stir to dissolve, then place in an ice bath, add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.13g, 0.35mmol) and 2,4,6-trimethylpyridine (0.26g, 2.15mmol), maintain the mixture at 0-5°C, and react for 1 hour with stirring. Add the reaction solution to HCl aq After adjusting the pH to 4-5 with 0.5N sodium hydroxide, the mixture was separated in a preparative liquid phase neutral water system to obtain 3 (70 mg), yield: 27%.

[0476] MS-ESI calculated value [M+H] + =1101, actual value 1101. 1 H NMR(400MHz,DMSO-d6)δ 8.65(t,J=6.8Hz,1H),8.60(d,J=8.8Hz,1H),8.34-8.24(m,2H),8.09(d,J=8.0Hz,1H),7.99(t,J=5.6Hz,1H),7 .81(d,J=11.2Hz,1H),7.79(s,1H),7.30-7.14(m,5H),7.04(s,2H),6.68(s,1H),5.91(d,J=16.8Hz,1H),5.70-5 .61(m,1H),5.56-5.45(m,2H),5.36(d,J=19.6Hz,1H),5.05(t,J=6.0Hz,1H),4.68(d,J=6.4Hz,2H),4.59(dd,J= 9.2,6.0Hz,1H),4.52-4.42(m,1H),4.17-4.05(m,2H),4.00-3.92(m,1H),3.89-3.79(m,1H),3.78-3.68(m,5H), 3.64-3.54(m,2H),3.01(dd,J=8.8,4.4Hz,1H),2.74(dd,J=13.6,9.6Hz,1H ),2.39(s,3H),2.26-2.18(m,2H),1.94-1.82(m,2H),0.86(t,J=7.6Hz,3H).

[0477] Example 1.4

[0478] [ka]

[0479] Phase 1 4a (10.00 g, 95.20 mmol) was dissolved in acetonitrile (100 mL), t-butyldimethylsilyl chloride (15.06 g, 99.92 mmol) was added, the mixture was cooled to 0 °C, and 1,8-diazobispiro[5.4.0]undecan-7-ene (13.73 g, 90.21 mmol) was added dropwise. After the addition was complete, the reaction mixture was allowed to return to room temperature and react with stirring for 16 hours. The product gradually precipitated. The reaction mixture was directly filtered, and the filter cake was collected to give 4b (11.50 g), yield: 55%.

[0480] MS-ESI calculated value [M+H] + =220, actual value 220. Phase 2 4b (5.55 g, 25.33 mmol), acetone (55.5 mL), and maleic anhydride (2.48 g, 25.33 mmol) were added sequentially to the reaction bottle. After the addition was complete, the mixture was stirred at room temperature for 2 hours. The reaction mixture was directly spin-dried to obtain 8.25 g of a yellow oil. The yellow oil was dissolved in toluene (82 mL), triethylamine (5.12 g, 50.66 mmol) was added, and the mixture was heated to 120 °C and refluxed for 2 hours. The reaction mixture was directly concentrated under reduced pressure to obtain a residue. The residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to obtain 4c (1.53 g) in a 33% yield.

[0481] MS-ESI calculated value [MH]-=184, found value 184. 1 H NMR(400MHz,DMSO-d6)δ 7.00(s,2H),6.05(s,1H),4.30(dd,J=9.6,5.6Hz,1H),3.98(dd,J=10.8,5.6Hz,1H),3.84(dd,J=10.8,10.8Hz,1H).

[0482] Phase 3 Add 4c (65 mg, 0.234 mmol), 2m (0.20 g, 0.23 mmol), and N,N-dimethylformamide (4 mL) to a reaction bottle in sequence, stir to dissolve, then place in an ice bath. Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.13 g, 0.35 mmol) and 2,4,6-trimethylpyridine (85 mg, 0.70 mmol), maintain the mixture at 0-5°C, and react for 1 hour with stirring. Add the reaction solution to HCl. aq After adjusting the pH to 4-5 with 0.5N sodium hydroxide, the compound was prepared and separated by a preparative liquid phase neutral water system to obtain 4 (70 mg), yield: 14%.

[0483] MS-ESI calculated value [M+H] + =1024, actual measured value 1024. 1 H NMR (400 MHz, DMSO-d6) δ 8.65(t,J=6.8Hz,1H),8.60(d,J=8.8Hz,1H),8.29(q,J=5.6Hz,2H),8.09(d,J =8.0Hz,1H),7.99(t,J=5.6Hz,1H),7.81(d,J=11.2Hz,1H),7.80(s,1H),7.29 -7.14(m,5H),7.04(s,2H),6.68(s,1H),5.91(d,J=16.8Hz,1H),5.71-5.60(m ,1H),5.56-5.45(m,2H),5.36(d,J=19.6Hz,1H),5.05(t,J=5.6Hz,1H),4.68(d ,J=6.4Hz,2H),4.59(dd,J=9.2,6.0Hz,1H),4.52-4.43(m,1H),4.17-4.04(m, 2H),4.00-3.92(m,1H),3.98-3.80(m,1H),3.78-3.68(m,5H),3.64-3.54(m,2 H),3.20-3.09(m,1H),3.00(dd,J=14.0,4.4Hz,1H),2.73(dd,J=13.6,9.6Hz, 1H),2.39(s,3H),2.26-2.17(m,2H),1.94-1.83(m,2H),0.86(t,J=7.2Hz,3H).

[0484] Example 1.5

[0485] [ka]

[0486] Phase 1 5a (650 mg, 1.70 mmol) and 5b (699 mg, 1.70 mmol) were dissolved in anhydrous dichloromethane (6 mL). 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (771 mg, 2.03 mmol) and N,N-diisopropylethylamine (440 mg, 2.40 mmol) were added sequentially in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction mixture was directly concentrated to give the residue, which was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 5c (1.10 g) in 83% yield.

[0487] MS-ESI calculated value [M+H] + =777, actual value 777. Phase 2 Dissolve 5c (1.10 g, 1.42 mmol) in anhydrous dichloromethane (2.6 mL) and add trifluoroacetic acid (2 mL) dropwise in an ice bath. After the addition is complete, return the reaction mixture to room temperature and allow to react for 2 hours with stirring. The reaction mixture is directly concentrated to give a residue, which is then separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give crude product 5d (1.16 g).

[0488] MS-ESI calculated value [M+H] + =721, actual value 721. Phase 3 5f (800 mg, 1.69 mmol) was dissolved in anhydrous acetonitrile (12 mL), and the solution was stirred for 1 hour at 40°C for 1 hour at 40°C for 1 hour at 40°C for 1 hour at 40°C for 1 minute ... After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 1 hour. 5e (750 mg, 1.69 mmol) was added to the reaction mixture, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (771 mg, 2.03 mmol) and N,N-diisopropylethylamine (654 mg, 5.07 mmol) were added sequentially in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 2 hours. The reaction mixture was diluted with ethyl acetate (200 mL), and the organic phase was washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to obtain crude product 5g (440 mg) in a 38% yield.

[0489] MS-ESI calculated value [M+Na] + =701, actual value 701. Phase 4 5g (420 mg, 0.62 mmol) was dissolved in anhydrous acetonitrile (10 mL) and 1,8-diazobispiro[5.4.0]undecan-7-ene (94 mg, 0.62 mmol) was added dropwise in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 2 hours. 5d (446 mg, 0.62 mmol) was added to the reaction mixture, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (353 mg, 0.93 mmol) and N,N-diisopropylethylamine (240 mg, 1.86 mmol) were added sequentially in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 1 hour. The reaction mixture was diluted with ethyl acetate (100 mL), and the organic phase was washed with saturated brine (25 mL × 3), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to give a residue, which was then purified by silica gel column chromatography (MeOH:DCM = 0 to 100%) to give 5h (600 mg) in a yield of 83%.

[0490] MS-ESI calculated value [M+Na] +=1181, actual value 1181. Stage 5 5h (600 mg, 0.52 mmol) was dissolved in a tetrahydrofuran / water (8 mL / 2 mL) mixture, Pd / C (60 mg, 10% w / w) was added, and the mixture was stirred under a hydrogen atmosphere for 5 h. The reaction mixture was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to give a residue. The residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 5i (360 mg) in a 65% yield.

[0491] MS-ESI calculated value [M+Na] + =1091.5, actual value 1091.8. Stage 6 1d (50 mg, 0.11 mmol), 5i (140 mg, 0.13 mmol), and N,N-dimethylformamide (2 mL) were added to a reaction bottle in this order, and after stirring to dissolve, the bottle was placed in an ice bath and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (65 mg, 0.17 mmol) and N,N Diisopropylethylamine (44 mg, 0.34 mmol) was added, and the mixture was stirred at 0-5°C for 1 hour. The reaction mixture was concentrated under reduced pressure to give a residue, which was then separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give crude product 5j (140 mg) in a yield of 85%.

[0492] MS-ESI calculated value [M+H] + =1502.6, actual value 1503. Stage 7 5j (55 mg, 0.037 mmol) was dissolved in anhydrous acetonitrile (2 mL), and diethylamine (27 mg, 0.37 mmol) was added dropwise in an ice bath. After the addition was complete, the reaction mixture was returned to room temperature and allowed to react overnight with stirring. The reaction mixture was concentrated under reduced pressure to obtain a residue. The product was separated by silica gel column chromatography (MeOH:DCM=0-100%) to give 5k (30 mg), yield: 63%.

[0493] MS-ESI calculated value [M+H] + =1280.6, actual value 1281. Stage 8 5k (30 mg, 0.023 mmol) and 5l (20 mg, 0.12 mmol) were dissolved in anhydrous dichloromethane (1 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (35 mg, 0.092 mmol) and N,N-diisopropylethylamine (16 mg, 0.12 mmol) were added in an ice bath. The mixture was stirred at room temperature for 1 hour. The reaction solution was then dissolved in HCl. aq After adjusting the pH to 4-5 with 0.5 N HCl, the compound was prepared and separated by a preparative liquid phase neutral water system to obtain 5 (15 mg), yield: 45%.

[0494] MS-ESI calculated value [M+H] + =1431.6, actual value 1432. Example 1.6

[0495] [ka]

[0496] Phase 1 Dissolve 6a (4.00 g, 21.03 mmol) in anhydrous dichloromethane (64 mL) and add Dess-Martin oxidant (11.15 g, 26.30 mmol). After the addition is complete, the mixture is stirred at room temperature for 2 h. Dilute the reaction mixture with dichloromethane (100 mL). The organic phase is washed sequentially with saturated sodium thiosulfate solution (50 mL × 1) and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure to give crude product 6b (5.30 g).

[0497] Phase 2 Crude products 6b (500 mg) and 6c (1.20 g, 3.19 mmol) were dissolved in anhydrous methanol (25 mL), and acetic acid (7.56 g, 126.00 mmol) and sodium cyanoborohydride (418 mg, 6.65 mmol) were added. After the addition was complete, the mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with dichloromethane (150 mL), and the excess acetic acid was quenched with saturated sodium bicarbonate (50 mL). The organic phase was separated. The residue was washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 6d (1.38 g).

[0498] MS-ESI calculated value [M+H] + =556, actual value 556. Phase 3 Crude product 6d (1.38 g) was dissolved in anhydrous tetrahydrofuran (30 mL), and N,N-diisopropylethylamine (482 mg, 3.73 mmol) and 9-fluorenylmethyl chloroformate (1.29 g, 4.97 mmol) were added. After the addition was complete, the mixture was stirred at room temperature for 3 h. The reaction mixture was diluted with dichloromethane (150 mL). The organic phase was washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (MeOH:DCM = 0-100%) to give 6e (1.00 g) in 65% yield.

[0499] MS-ESI calculated value [M+H] + =778, actual value 778. Phase 4 6e (1.00 g, 1.28 mmol) was dissolved in anhydrous dichloromethane (4 mL), trifluoroacetic acid (1.46 g, 12.85 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After that, trifluoroacetic acid (1.46 g, 12.85 mmol) was added, and the reaction mixture was concentrated under reduced pressure to give a residue. The residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 6f (500 mg) in a 54% yield.

[0500] MS-ESI calculated value [M+H] + =722.4, actual value 722.9. Stage 5 2j (500 mg, 0.68 mmol) was dissolved in anhydrous acetonitrile (5 mL), and 1,8-diazobispiro[5.4.0]undecan-7-ene (134 mg, 0.88 mmol) was added dropwise in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 2 hours. 6f (490 mg, 0.68 mmol) was added to the reaction mixture, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (388 mg, 1.02 mmol) and N,N-diisopropylethylamine (220 mg, 1.70 mmol) were added sequentially in an ice bath. After the addition was complete, the reaction mixture was allowed to warm to room temperature and reacted with stirring for 1 hour. The reaction mixture was diluted with dichloromethane (100 mL), and the organic phase was washed with saturated brine (25 mL × 3), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was separated by silica gel column chromatography (MeOH:DCM = 0 to 100%) to obtain 6 g (470 mg), yield: 57%.

[0501] MS-ESI calculated value [M+Na] + =1239.6, actual value 1239.9. 1 H NMR(400MHz,DMSO-d6)δ 7.84(d,J=7.6Hz,2H),7.82-7.74(m,1H),7.63(d,J=7.6Hz,2H),7.61-7.52(m,2H),7.50- 7.44(m,2H),7.44-7.24(m,15H),4.90-4.81(m,2H),4.73-4.62(m,1H),4.59(m,J=5.6Hz,2 H),4.30(s,2H),4.29-4.25(m,1H),4.15-4.02(m,1H),4.00-3.89(m,2H),3.86(d,J=5.2H z,2H),3.80-3.45(m,34H),3.43(s,3H),3.41-3.38(m,1H),3.11(dd,J=14.0,10.0Hz,1H).

[0502] Stage 6 6g (470 mg, 0.39 mmol) was dissolved in a tetrahydrofuran / water (4 mL / 1 mL) mixture, wet Pd / C (62 mg, 13% w / w) was added, and the mixture was stirred under a hydrogen atmosphere for 2 h. The reaction mixture was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to give crude product 6h (360 mg).

[0503] MS-ESI calculated value [M+Na] + =1149.5, actual value 1149.9. Referring to Example 5, compound 5 is synthesized from compound 5i and compound 1d through three reaction steps, and compound 6 is synthesized from compound 6h and compound 1d through three reaction steps.

[0504] MS-ESI calculated value [M+H] + =1489.6, actual value 1490. Example 1.7

[0505] [ka]

[0506] Synthetic Route 1

[0507] [ka]

[0508] Phase 1 Under a nitrogen atmosphere, 7a (10.60 g, 30.00 mmol) was dissolved in a tetrahydrofuran / toluene (200 mL / 50 mL) mixed solvent, and lead tetraacetate (17.30 g, 39.00 mmol) and pyridine (3.08 g, 39.00 mmol) were added. The mixture was heated to 75 °C and stirred for 4 hours. The reaction mixture was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to remove most of the solvent. The residue was dissolved in ethyl acetate (400 mL). The organic phase was washed successively with water (100 mL × 2), saturated aqueous sodium bicarbonate (100 mL), dilute hydrochloric acid (0.5 N, 100 mL × 2), and saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography. Separation by column chromatography (EA:hexane=0-100%) gave 7b (9.70 g), yield: 88%.

[0509] MS-ESI calculated value [M+Na] + =391, actual value 391. Phase 2 Under a nitrogen atmosphere, 7b (8.00 g, 21.74 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). p-Toluenesulfonic acid (374 mg, 2.18 mmol) and t-butylglycolic acid (8.60 g, 65.14 mmol) were added sequentially in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 1 h. The reaction mixture was diluted with ethyl acetate (200 mL). The organic phase was washed sequentially with water (50 mL × 2), saturated aqueous sodium bicarbonate (50 mL), and saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EA: hexane = 0-100%) to give 7c (5.90 g) in a 62% yield.

[0510] MS-ESI calculated value [M+Na] + =463, actual value 463. Phase 3 Under a nitrogen atmosphere, 7c (4.90 g, 11.14 mmol) was dissolved in anhydrous acetonitrile (40 mL). 1,8-diazobispiro[5.4.0]undecan-7-ene (847 mg, 5.57 mmol) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 3 hours. 7d (6.13 g, 11.14 mmol) was added to the reaction mixture. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (6.34 mg, 16.68 mmol) and N,N-diisopropylethylamine (2.88 g, 22.32 mmol) were added sequentially in an ice bath. After the addition was complete, the reaction was allowed to warm to room temperature and react with stirring for 1 hour. The reaction mixture was diluted with ethyl acetate (200 mL), and the organic phase was washed with saturated brine (25 mL × 3), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to give a residue, which was then purified by silica gel column chromatography (MeOH:DCM = 0-100%) to give 7e (5.8 g), yield: 74%.

[0511] MS-ESI calculated value [M+Na] + =724, actual value 724. Phase 4 Under a nitrogen atmosphere, 7e (2.80 g, 3.99 mmol) was dissolved in anhydrous acetonitrile (30 mL). 1,8-diazobispiro[5.4.0]undecan-7-ene (606 mg, 3.99 mmol) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 5 hours. The reaction mixture was cooled to 0 °C, and 1-hydroxybenzotriazole (1.08 g, 7.98 mmol) was added in batches. A solid (jelly-like) precipitated from the reaction mixture. The mixture was stirred in an ice bath for 1 hour, then filtered. The top filter cake was slurried in isopropyl ether / n-hexane (10 mL / 40 mL) and filtered to give 7f (2.0 g).

[0512] MS-ESI calculated value [M+H] + =480, actual value 480. Stage 5 7g (0.99 g, 3.20 mmol) and 7f (2.40 g, 3.20 mmol) were dissolved in a mixed solvent of acetonitrile / water (15 mL / 15 mL), and 2,4,6-trimethylpyridine (387 mg, 3.20 mmol) was added in an ice bath. After the addition was complete, the mixture was returned to room temperature and reacted with stirring for 2 hours. The reaction solution was diluted with ethyl acetate (200 mL), and the organic phase was washed successively with water (30 mL x 2), dilute hydrochloric acid (0.5 N, 50 mL x 2), and saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (MeOH:DCM = 0.05). 100%) to give 7h (1.25 g), yield: 58%.

[0513] MS-ESI calculated value [M+Na] + =697, actual value 697. Stage 6 Under a nitrogen atmosphere, zinc bromide (8.00 g, 35.60 mmol) was weighed into a reaction bottle, nitromethane (15 mL) was added, and the mixture was stirred at room temperature for 10 minutes until an emulsion formed. The reaction bottle was placed in an ice bath, and a solution of 7i (1.20 g, 1.78 mmol) in nitromethane (20 mL) was added dropwise. After the addition was complete, the mixture was maintained in the ice bath and allowed to react for 4 hours with continuous stirring. The reaction mixture was diluted with water (50 mL), and the aqueous phase was adjusted to pH 5-6 with aqueous ammonia (2% w / w) and separated. The aqueous phase was washed with dichloromethane (50 mL x 3) and then lyophilized. The lyophilized residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to give 7i (450 mg) in a 41% yield.

[0514] MS-ESI calculated value [M+Na] + =641, actual value 641. 1H NMR(400MHz,DMSO-d6)δ 7.28-7.14(m,5H),6.97(s,2H),4.66-4.54(m,2H),4.43(dd,J=9.2,4.4Hz,1H),3.81(s,2H),3.77 -3.52(m,12H),3.07(dd,J=14.0,4.8Hz,1H),2.81(dd,J=13.6,9.6Hz,1H),2.33(t,J=6.0Hz,2H).

[0515] Stage 7 Compound 1d (50 mg, 0.11 mmol), 7i (74 mg, 0.12 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (31 mg, 0.16 mmol), and N,N-dimethylformamide (2 mL) were added sequentially to a 10 mL single-bottle mixture. 2,4,6-Trimethylpyridine (60 mg, 0.49 mmol) was added dropwise to the mixture in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react for 3 hours with stirring. The reaction mixture was added dropwise to methyl t-butyl ether (60 mL), and a solid precipitated. The solid was collected by filtration, dissolved in a dichloromethane / methanol mixture (24 mL / 4 mL), and concentrated to give the crude product. The crude product was purified by liquid-phase preparative separation to give compound 7 (16 mg) in a 17% yield.

[0516] MS-ESI calculated value [M+H] + =1052, measured value: 1052. 1H NMR (400MHz, DMSO-d6)δ 8.65(t,J=6.8Hz,1H),8.61(d,J=9.2Hz,1H),8.28(t,J=4.4Hz,1H),8.14-8.06 (m,2H),8.06(t,J=5.6Hz,1H),7.82(d,J=11.2Hz,1H),7.79(s,1H),7.28-7.13( m,5H),6.99(s,2H),5.92(d,J=16.8Hz,1H),5.70-5,60(m,1H),5.53(d,J=8.0H z,1H),5.48(d,J=4.8Hz,1H),5.37(d,J=19.6Hz,1H),4.68(d,J=6.4Hz,1H),4.5 1-4.41(m,1H),4.17-4.04(m,2H),3.72-3.68(m,2H),3.68-3.63(m,2H),3.62- 3.58(m,1H),3.58-3.54(m,3H),3.54-3.39(m,3H),3.48-3.43(m,2H),3.19-3.0 8(m,1H),3.06-2.97(m,1H),2.80-2.70(m,1H),2.70-2.65(m,1H),2.39(s,3H), 2.35-2.28(m,3H),2.27-2.18(m,2H),1.94-1.82(m,2H),0.85(t,J=7.2Hz,3H).

[0517] Synthesis Road 2

[0518]

change

[0519] Stage 1 Under a nitrogen atmosphere, 8a (4.50 g, 8.98 mmol) was dissolved in anhydrous N,N-dimethylformamide (25 mL), and acetic acid (1.19 g, 19.76 mmol), lead tetraacetate (7.96 g, 17.96 mmol), and copper acetate (1.10 g, 8.98 mmol) were added. The mixture was heated to 50 °C and stirred for 30 min. The reaction mixture was quenched by adding water (50 mL). The aqueous phase was extracted with ethyl acetate (500 mL). The organic phase was washed sequentially with saturated aqueous sodium bicarbonate (100 mL × 1), water (100 mL), and saturated brine (50 mL × 4), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give crude product 8b (4.70 g).

[0520] MS-ESI calculated value [M+Na] + =538, actual value 538. Phase 2 Under a nitrogen atmosphere, 8b (4.70 g, 9.13 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). p-Toluenesulfonic acid (235 mg, 1.37 mmol) and benzyl glycolic acid (5.30 g, 31.96 mmol) were added sequentially in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 4 hours. The reaction mixture was diluted with ethyl acetate (200 mL). The organic phase was washed sequentially with water (50 mL × 1), saturated aqueous sodium bicarbonate (50 mL × 1), and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EA: hexane = 0-100%) to give 8c (4.50 g) in a 79% yield.

[0521] MS-ESI calculated value [M+Na] + =644, actual value 644. Phase 3 8c (4.50 g, 7.25 mmol) was dissolved in a tetrahydrofuran / water (50 mL / 15 mL) mixture, wet Pd / C (380 mg, 8% w / w) was added, and the mixture was stirred overnight under a hydrogen atmosphere. The reaction mixture was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to give crude product 8d (2.65 g).

[0522] MS-ESI calculated value [M+Na] + =554, actual value 554. Phase 4 Under a nitrogen atmosphere, 8d (2.65 g, 4.99 mmol) was dissolved in anhydrous acetonitrile (30 mL), and 1,8-diazobispyro[5.4.0]undecane-7-ene (1.52 g, 9.98 mmol) was added in an ice bath. After the addition was completed, the mixture was returned to room temperature and stirred. The reaction mixture was cooled to 0°C, and 1-hydroxybenzotriazole (1.35 g, 9.98 mmol) was added in batches. A solid precipitated from the reaction mixture. The mixture was stirred in an ice bath for 2 hours, and then filtered. The filter cake was collected and dried under suction to give crude product 8e (1.85 g).

[0523] MS-ESI calculated value [M+H] + =310, actual value 310. Stage 5 7g (3.10 g, 10.00 mmol) and 8f (1.45 g, 11.00 mmol) were dissolved in a mixture of acetonitrile and water (25 mL / 8 mL). Triethylamine (1.51 g, 15.00 mmol) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react for 2 hours with stirring. Most of the acetonitrile was removed by spinning. The residue was diluted with dichloromethane and water (100 mL / 50 mL), separated, and the aqueous phase was washed with dichloromethane (50 mL x 2) and lyophilized. After lyophilization, the residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to obtain crude product 8g (4.25 g).

[0524] MS-ESI calculated value [M+H] + =328, actual value 328. Stage 6 8g (1.03 g) and N-hydroxysuccinimide (552 mg, 4.80 mmol) were dissolved in anhydrous dichloromethane (10 mL). Dicyclohexylcarbodiimide (0.99 g, 4.80 mmol) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 1.5 hours. The reaction mixture was filtered to remove insoluble solids, and the mother liquor was reserved. 8e (0.52 g) and 2,4,6-trimethylpyridine (203 mg, 1.68 mmol) were added to the mother liquor in an ice bath. The mixture was allowed to warm to room temperature and react with stirring for 4 hours. The reaction mixture was diluted with water (50 mL). The aqueous phase was washed with dichloromethane (50 mL x 2) and then lyophilized. After lyophilization, the residue was separated by silica gel column chromatography (MeOH:DCM = 0-100%) to obtain 7i (630 mg).

[0525] MS-ESI calculated value [M+Na] + =641, actual value 641. Stage 7 Compounds 1d (110 mg, 0.24 mmol) and 7i (195 mg, 0.32 mmol) were added sequentially to a 10 mL single-bottle flask. Anhydrous N,N-dimethylformamide (3 mL) was added, and the mixture was cooled to 0 °C. Trifluoroacetic acid (29 mg, 0.25 mmol), 2,4,6-trimethylpyridine (133 mg, 1.10 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (68 mg, 0.35 mmol) were added sequentially. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 3 h. The reaction mixture was directly purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 7 (86 mg) in a 34% yield.

[0526] MS-ESI calculated value [M+H] + =1052, actual value 1052. Synthetic Route 3

[0527] [ka]

[0528] Phase 1 Under a nitrogen atmosphere, 9a (5.10 g, 11.48 mmol) was dissolved in anhydrous dichloromethane (100 mL) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (6.54 g, 17.22 mmol), N,N-diisopropylethylamine (4.44 g, 34.44 mmol), and glycine t-butyl ester (1.80 g, 13.78 mmol) were added sequentially. After the addition was complete, the mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with dichloromethane (100 mL). The organic phase was washed sequentially with dilute hydrochloric acid (2 N, 50 mL × 1), water (50 mL × 1), and saturated brine (50 mL × 2), dried, filtered, and the filtrate was concentrated under reduced pressure to give crude white solid 9b (6.80 g).

[0529] MS-ESI calculated value [M+H] + =558, actual value 558. Phase 2 Under a nitrogen atmosphere, the crude product 9b (6.30 g) from the previous step was dissolved in anhydrous acetonitrile (100 mL). 1,8-Diazacyclo[5,4,0]undecene-7 (3.44 g, 22.62 mmol) was added in an ice bath. The mixture was allowed to warm to room temperature and react with stirring for 5 hours. The reaction bottle was then placed back in the ice bath, and Fmoc-glycine (4.03 g, 13.57 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.16 g, 13.57 mmol), and N,N-diisopropylethylamine (2.19 g, 16.97 mmol) were added sequentially. After the addition was complete, the mixture was allowed to react at room temperature with continued stirring for 1 hour. The reaction mixture was diluted with ethyl acetate (200 mL). The organic phase was washed successively with dilute hydrochloric acid (2N, 50 mL × 1), water (50 mL × 2), and saturated brine (50 mL × 2), then dried and filtered. The filtrate was concentrated under reduced pressure to give a residue. The residue was separated by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to give crude product 9c (8.5 g).

[0530] MS-ESI calculated value [M+H] + =615, actual value 615. Phase 3 Under a nitrogen atmosphere, 9c (8.50 g, 13.83 mmol) was dissolved in dichloromethane (20 mL), cooled to 0 °C in an ice-water bath, and trifluoroacetic acid (20 mL, 0.261 mol) was added dropwise. The mixture was allowed to warm to room temperature and react for 4 hours with stirring. The mixture was concentrated under reduced pressure, and ethyl acetate (100 mL) and water (100 mL) were added to the residue. The pH of the solution was adjusted to 4-5 with saturated sodium bicarbonate (a solid precipitated, forming an aqueous emulsion). The mixture was separated, the aqueous emulsion was collected, and the aqueous phase was washed with ethyl acetate (50 mL × 2). The aqueous phase was washed, adjusted to pH 1-2 with 1N aqueous hydrochloric acid, dichloromethane (200 mL) was added, the layers were separated, the aqueous phase was extracted with dichloromethane (150 mL × 2), the organic phases were combined, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product 9d (4.10 g) as a white solid.

[0531] MS-ESI calculated value [M+H] + =559, actual value 559. Phase 4 Under a nitrogen atmosphere, 9d (4.10 g, 7.34 mmol), acetic acid (1.78 g, 29.35 mmol), lead tetraacetate (13.00 g, 17.96 mmol), and copper acetate (1.80 g, 14.68 mmol) were sequentially added to a reaction bottle. The mixture was heated to 50 °C and stirred for 30 min. The reaction mixture was quenched by adding water (50 mL). The aqueous phase was extracted with dichloromethane (500 mL). The organic phase was washed sequentially with saturated aqueous sodium bicarbonate (100 mL × 1), water (100 mL), and saturated brine (50 mL × 4), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give crude product 9e (3.84 g).

[0532] MS-ESI calculated value [M+Na] + =595, actual value 595. Stage 5 Under a nitrogen atmosphere, 9e (3.80 g, 6.60 mmol) was dissolved in anhydrous tetrahydrofuran (40 mL). p-Toluenesulfonic acid (171 mg, 1.00 mmol) and benzyl glycolic acid (3.86 g, 25.20 mmol) were added sequentially in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 4 hours. The reaction mixture was diluted with ethyl acetate (200 mL). The organic phase was washed sequentially with water (50 mL × 1), saturated aqueous sodium bicarbonate (50 mL × 1), and saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give 9f (2.14 g) in a 48% yield.

[0533] MS-ESI calculated value [M+Na] + =701, actual value 701. Stage 6 9f (2.10 g, 3.09 mmol) was dissolved in a tetrahydrofuran / water (50 mL / 15 mL) mixture, wet Pd / C (168 mg, 8% w / w) was added, and the mixture was stirred under a hydrogen atmosphere for 5 h. The reaction mixture was filtered to remove insoluble materials, and the filtrate was concentrated under reduced pressure to give crude product 9f (2.40 g).

[0534] MS-ESI calculated value [M+Na] + =611, actual value 611. Stage 7 Under a nitrogen atmosphere, 9g (2.18 g, 3.70 mmol) was dissolved in anhydrous acetonitrile (30 mL). 1,8-diazobispiro[5.4.0]undecan-7-ene (1.13 g, 7.41 mmol) was added in an ice bath. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 5 hours. The reaction mixture was cooled to 0 °C, and 1-hydroxybenzotriazole (1.35 g, 9.98 mmol) was added in batches. A solid precipitated from the reaction mixture. Acetonitrile (30 mL) was added, and the mixture was slurried at room temperature for 2 hours. The mixture was filtered, washed with acetonitrile (2 x 20 mL), and dried under suction to give crude 9h (1.83 g) as a white powder.

[0535] MS-ESI calculated value [M+H] + =367, actual value 367. Stage 8 9i (500 mg, 1.43 mmol, 9i:2,4,6-trimethylpyridine = 1:0.655), 2,4,6-trimethylpyridine (494 mg, 4.08 mmol), and N-Hydroxysuccinimide (206 mg, 1.79 mmol) was dissolved in anhydrous dichloromethane (25 mL) and dicyclohexylcarbodiimide (443 mg, 2.15 mmol) was added in an ice bath. After the addition was complete, the mixture was stirred at 0 °C for 3 h. 9h (600 mg, 1.60 mmol) was added to the reaction mixture in an ice bath, and the mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was diluted with water (100 mL). The aqueous phase was washed with dichloromethane (100 mL x 3). The aqueous phase was lyophilized. After lyophilization, the residue was separated by silica gel column chromatography (MeOH:DCM = 0-50%) to give 7i (550 mg) in a 62% yield.

[0536] MS-ESI calculated value [M+Na] + =641, measured value 641. Stage 9 Compounds 1d (110 mg, 0.24 mmol) and 7i (195 mg, 0.32 mmol) were added sequentially to a 10 mL single-bottle flask. Anhydrous N,N-dimethylformamide (3 mL) was added, and the mixture was cooled to 0 °C. Trifluoroacetic acid (29 mg, 0.25 mmol), 2,4,6-trimethylpyridine (133 mg, 1.10 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (68 mg, 0.35 mmol) were added sequentially. After the addition was complete, the mixture was allowed to warm to room temperature and react with stirring for 3 h. The reaction mixture was directly purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 7 (90 mg) in a 35% yield.

[0537] MS-ESI calculated value [M+H] + =1052, actual value 1052. Example 1.8

[0538] [ka]

[0539] Phase 1 Under nitrogen gas protection, 10a (10 g, 22.91 mmol) and N6-Boc-L-lysine t-butyl ester hydrochloride (9.32 g, 27.50 mmol) were dissolved in a tetrahydrofuran / water mixed solvent (V テトラヒドロフラン :V 水 = 4:1, 100 mL) and mixed The mixture was cooled to 0°C and stirred for 10 minutes, after which sodium bicarbonate (3.85 g, 45.83 mmol) was added, and the mixture was stirred and reacted for 2 hours at 0°C. Water (100 mL) and ethyl acetate (100 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (100 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give compound 10b (12.50 g) in an 87% yield.

[0540] MS-ESI calculated value [M+Na] + 646, actual value 646. Phase 2 10b (12.50 g, 20.04 mmol) was dissolved in 4M hydrogen chloride in dioxane (63 mL), and the mixture was stirred at room temperature for 3 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was slurried and purified with methyl t-butyl ether (150 mL) to give crude product 10c (11.40 g).

[0541] MS-ESI calculated value [M+H] + =468, actual value 468. Phase 3 10c (11.40 g) was dissolved in anhydrous dichloromethane (190 mL), propionaldehyde (7.01 g, 120.70 mmol) was added, and the mixture was allowed to react at room temperature with stirring for 15 minutes, after which sodium triacetoxyborohydride (21.33 g, 100.64 mmol) was added in batches, and after the addition was complete the mixture was allowed to react at room temperature with continued stirring for 1 hour. The reaction mixture was quenched with saturated aqueous ammonium chloride (10 mL), stirred at room temperature for 1 hour, added with water (200 mL) and ethyl acetate (100 mL), and the mixture was separated. The aqueous phase was extracted with ethyl acetate (100 mL × 2). The organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was then concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0~100%) to give compound 10d (9.40 g). The two-step yield: 85%.

[0542] MS-ESI calculated value [M+H] + =552, actual value 552. Phase 4 Under nitrogen gas protection, 10e (2.5 g, 5.27 mmol) was dissolved in anhydrous dichloromethane (35 mL), 1,8-diazobispiro[5.4.0]undecan-7-ene (1.20 g, 7.88 mmol) was added, and the mixture was cooled to 0 °C and stirred for 10 min, then returned to room temperature and stirred for 1 h. The mixture was cooled again to 0 °C, and 10d (3.49 g, 6.33 mmol), N,N-diisopropylethylamine (1.02 g, 7.89 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.01 g, 7.92 mmol) were added. After the addition was complete, the mixture was maintained at 0 °C and stirred for 1 h. Water (50 mL) and ethyl acetate (50 mL) were added to the reaction mixture, and the layers were separated. The aqueous phase was extracted with ethyl acetate (100 mL × 2). The organic phases were combined and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to give compound 10f (2.00 g) in a 48% yield.

[0543] MS-ESI calculated value [M+H] + =786, actual value 786. Stage 5 10f (2.00 g, 2.54 mmol) was dissolved in a tetrahydrofuran / water mixture (V テトラヒドロフラン :V 水 The mixture was dissolved in 10% wet palladium on carbon (200 mg) in a 3:1 ratio (40 mL), and the mixture was stirred at room temperature under a hydrogen gas atmosphere for 3 hours. The reaction mixture was filtered to remove insoluble matter, and the filter cake was eluted with ethyl acetate. The filtrate was then evaporated under reduced pressure. The mixture was concentrated under reduced pressure to obtain the residual aqueous phase, which was then extracted with ethyl acetate (20 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was then filtered and the filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to obtain compound 10g (930 mg), yield: 53%.

[0544] MS-ESI calculated value [M+H] + =696, actual value 696. Stage 6 Under a nitrogen atmosphere, 10g (652 mg, 0.94 mmol) and 1d (330 mg, 0.73 mmol) were dissolved in anhydrous dichloromethane (5 mL). The mixture was cooled to 0 °C, and N,N-diisopropylethylamine (189 mg, 1.46 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (330 mg, 0.87 mmol) were added. The mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 10h (730 mg) in 88% yield.

[0545] MS-ESI calculated value [M+H] + =1129, actual value 1129. Stage 7 Under nitrogen gas protection, 10h (720 mg, 0.64 mmol) was dissolved in anhydrous tetrahydrofuran (7 mL), cooled to 0 °C, piperidine (272 mg, 3.19 mmol) was added, and the mixture was stirred at 0 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to give compound 10i (350 mg) in a 61% yield.

[0546] MS-ESI calculated value [M+H] + =907, actual value 907. 1 H NMR(400MHz,DMSO-d6)δ 7.78(s,1H),7.72(d,J=10.8Hz,1H),5.91(d,J=16.4Hz,1H),5.70-5.60(m,1H),5.49(d,J=16.4Hz,1H),4.7 4-4.62(m,2H),4.24-4.15(m,1H),4.13(d,J=6.0Hz,2H),3.81-3.61(m,2H),3.34-3.20(m,1H),3.18(s,2H) ,3.16-3.07(m,1H),3.00(d,J=4.8Hz,1H),2.93-2.81(m,2H),2.37(s,3H),2.31-2.21(m,5H),1.99-1.83(m ,3H),1.67-1.46(m,4H),1.39-1.22(m,8H),0.91-0.82(m,6H),0.82(t,J=7.2Hz,6H),0.76(d,J=6.8Hz,3H).

[0547] Stage 8 10i (150 mg, 0.17 mmol) was dissolved in anhydrous N,N-dimethylformamide (2 mL), 2,4,6-trimethylpyridine (60 mg, 0.50 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (94 mg, 0.25 mmol) were added, and the mixture was stirred at 0 °C for 5 minutes. 4c (57 mg, 0.20 mmol) was then added, and the mixture was stirred at 0 °C for 1 hour. Water (5 mL) and ethyl acetate (10 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to give compound 10 (125 mg) in 67% yield.

[0548] MS-ESI calculated value [M+H] + =1074, actual value 1074. Example 1.9

[0549] [ka]

[0550] Phase 1 Under nitrogen gas protection, 11a (5.00 g, 12.18 mmol) and p-aminobenzyl alcohol (2.25 g, 18.27 mmol) were dissolved in anhydrous dichloromethane (100 mL). The mixture was cooled to 0°C, and then N,N-diisopropylethylamine (3.94 g, 30.49 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.56 g, 14.62 mmol) were added. After the addition was complete, the mixture was stirred at 0°C for 1 hour. The reaction mixture was then diluted with water (200 mL) and a dichloromethane / methanol mixed solvent (V). ジクロロメタン :V メタノール The aqueous phase was separated into 100 ml of a dichloromethane / methanol mixed solvent (V ジクロロメタン :V メタノールThe organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to give a residue, which was purified by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to give compound 11b (6.68 g), yield: 100%.

[0551] MS-ESI calculated value [M-OH] + =498, actual value 498. Phase 2 Under nitrogen gas protection, 11b (3.23 g, 6.26 mmol) and bis(4-nitrophenyl) carbonate (2.86 g, 9.40 mmol) were dissolved in a mixed solvent of anhydrous dichloromethane / N,N-dimethylformamide (V ジクロロメタン :V N、N-ジメチルホルムアミド The resulting mixture was dissolved in 10 mL of methyl tert-butyl ether (2:1, 48 mL), N,N-diisopropylethylamine (2.43 g, 18.80 mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction mixture was quenched with water (100 mL), ethyl acetate (100 mL) was added, and the layers were separated. The aqueous phase was extracted with ethyl acetate (100 mL x 2), and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was slurried with methyl tert-butyl ether (10 mL), and purified to give compound 11c (2.00 g), yield: 47%.

[0552] MS-ESI calculated value [M+Na] + =703, actual value 703. Phase 3 Under nitrogen gas protection, 11c (1.27 g, 1.87 mmol) and 1d (430 mg, 0.95 mmol) were dissolved in anhydrous N,N-dimethylformamide (12 mL). The mixture was cooled to 0°C, and then 3 drops of triethylamine were added. The mixture was maintained at 0°C and stirred for 15 minutes. Then, 1-hydroxybenzotriazole (299 mg, 2.21 mmol) and pyridine (2.15 g, 27.18 mmol) were added. After the addition was complete, the mixture was maintained at 0°C and stirred for 15 minutes. The mixture was then returned to room temperature and allowed to react for 2 hours with stirring. Water (20 mL) and ethyl acetate (30 mL) were added to the reaction mixture, and the layers were separated. The aqueous phase was extracted with ethyl acetate (30 mL × 2). The organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was then filtered and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0 to 100%) to give compound 11d (550 mg) in a 58% yield.

[0553] MS-ESI calculated value [M+H] + =993, actual measured value 993. Phase 4 11d (500 mg, 0.50 mmol) was dissolved in 1,4-dioxane (5 mL), piperidine (5 mL) was added, and the mixture was stirred at room temperature for 2 hours. Petroleum ether (5 mL) was added dropwise to the reaction mixture, and a large amount of solid precipitated. The solid was filtered, and the filter cake was collected and dried to give 11e (386 mg), yield: 94%.

[0554] MS-ESI calculated value [M+H] + =771, actual value 771. 1H NMR(400MHz,DMSO-d6)δ 7.76(s,1H),7.66(d,J=10.4Hz,1H),7.60(d,J=8.0Hz,2H),7.43(d,J=8.4Hz,2H),5.91(d,J=16.4Hz,1 H),5.49(d,J=16.4Hz,1H),5.29(q,J=4.8Hz,1H),5.20-5.02(m,2H),4.50-4.39(m,1H),3.30-3.14(m,1 H),3.14-3.02(m,1H),2.99(d,J=5.2Hz,1H),2.31(s,3H),2.30-2.21(m,1H),2.13-1.98(m,1H),1.98- 1.80(m,3H),1.30(d,J=7.2Hz,3H),0.88(d,J=6.8Hz,3H),0.84(t,J=7.2Hz,3H),0.78(d,J=6.8Hz,3H).

[0555] Stage 5 11e (150 mg, 0.20 mmol) was dissolved in anhydrous N,N-dimethylformamide (2 mL), 2,4,6-trimethylpyridine (71 mg, 0.59 mmol), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (111 mg, 0.29 mmol) were added, and the mixture was stirred at 0 °C for 5 min. 4c (67 mg, 0.23 mmol) was then added. After the addition was complete, the mixture was stirred at 0 °C for 1 h. Water (5 mL) and ethyl acetate (10 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to give compound 11 (38 mg) in 21% yield.

[0556] MS-ESI calculated value [M+H] + =938, actual measured value 938. 1H NMR(400MHz,DMSO-d6)δ 8.56-8.00(m,3H),7.78(s,1H),7.72(d,J=10.8Hz,1H),7.59(d,J=8.0Hz,2H),7.42(d,J=8.0Hz,2H),7.02(s,2H) ,5.92(d,J=16.4Hz,1H),5.65-5.42(m,2H),5.42-5.26(m,2H),5.23-4.97(m,2 H),4.68-4.51(m,1H),4.41-4.27(m,1H),4.23-4.11(m,1H),4.04-3.94(m,1H) ,3.92-3.77(m,1H),3.31-3.18(m,1H),3.16-2.92(m,1H),2.35(s,3H),2.30-2 .20(m,1H),2.15-2.02(m,1H),2.03-1.91(m,1H),1.92-1.82(m,2H),1.31-1.26 m,3H),0.86(t,J=6.8Hz,3H),0.83-0.78(m,3H),0.77(d,J=6.8Hz,3H).

[0557] Example 1.10

[0558] [ka]

[0559] Phase 1 12a (4.00 g, 6.65 mmol) and bis(4-nitrophenyl)carbonate (10.08 g, 33.14 mmol) were dissolved in anhydrous N,N-dimethylformamide (80 mL). N,N-diisopropylethylamine (2.40 g, 18.57 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Ethyl acetate (120 mL) and petroleum ether (240 mL) were added dropwise to the reaction solution. After the addition was complete, the mixture was stirred at room temperature for 10 minutes, then filtered. The filter cake was collected and dried to give compound 12b (3.30 g), 65% yield.

[0560] MS-ESI calculated value [M+H] + =767, actual value 767. Phase 2 Under nitrogen gas protection, 12b (675 mg, 0.88 mmol) and 1d (270 mg, 0.60 mmol) were dissolved in anhydrous N,N-dimethylformamide (5 mL). The mixture was cooled to 0°C, and then 3 drops of triethylamine were added. After stirring for 15 minutes, 1-hydroxybenzotriazole (189 mg, 1.40 mmol) and pyridine (1.36 g, 1 The mixture was stirred at 0°C for 15 minutes, then allowed to warm to room temperature and react for 2 hours with continuous stirring. Water (20 mL) and ethyl acetate (30 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (30 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 12c (350 mg) in a 54% yield.

[0561] MS-ESI calculated value [M+Na] + =1101, actual value 1101. Phase 3 12c (570 mg, 0.53 mmol) was dissolved in anhydrous tetrahydrofuran (10 mL), piperidine (450 mg, 5.28 mmol) was added, and the mixture was stirred at room temperature for 3 hours. Methyl t-butyl ether (15 mL) was added to the reaction mixture, and a solid precipitated. The mixture was stirred for 30 minutes, filtered, and the filter cake was dried to give compound 12d (400 mg) in a yield of 88%.

[0562] MS-ESI calculated value [M+H] + =857, actual value 857. Phase 4 12d (210 mg, 0.25 mmol) was dissolved in anhydrous N,N-dimethylformamide (3 mL), 2,4,6-trimethylpyridine (89 mg, 0.73 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (140 mg, 0.37 mmol) were added, and the mixture was stirred at 0 °C for 5 minutes. 4c (71 mg, 0.25 mmol) was then added, and the mixture was stirred at 0 °C for 1 hour. Water (5 mL) and ethyl acetate (10 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (10 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a residue, which was purified by preparative HPLC to give compound 12 (30 mg) in 12% yield.

[0563] MS-ESI calculated value [M+H] + =1024, actual measured value 1024. 1 H NMR(500MHz,DMSO-d6)δ 7.79(s,1H),7.75(d,J=10.5Hz,1H),7.59(d,J=7.5Hz,1H),7.42(d,J=8.5 Hz,1H),7.02(s,2H),5.92(d,J=17.0Hz,1H),5.57(d,J=20.0Hz,1H),5.51( d,J=16.5Hz,1H),5.37(d,J=19.5Hz,1H),5.33-5.28(m,1H),5.16(d,J=12 .5Hz,1H),5.08(d,J=12.5Hz,1H),4.60-4.54(m,1H),4.37-4.30(m,1H),4. 20-4.14(m,1H),4.00-3.93(m,1H),3.88-3.81(m,1H),3.30-3.21(m,1H), 3.15-3.06(m,1H),3.05-2.89(m,2H),2.36(s,3H),2.31-2.23(m,1H),2.14 -2.03(m,1H),2.02-1.93(m,1H),1.92-1.82(m,2H),1.74-1.64(m,1H),164 -1.53(m,1H),1.48-1.31(m,2H),0.89-0.80(m,6H),0.76(d,J=6.5Hz,3H).

[0564] Example 1.11

[0565] [ka]

[0566] Phase 1 12b (427 mg, 0.56 mmol) and 13a (270 mg, 0.38 mmol) were dissolved in anhydrous N,N-dimethylformamide (15 mL), 1-hydroxybenzotriazole (51 mg, 0.38 mmol), and pyridine (744 mg, 9.41 mmol) were added, and the mixture was stirred at room temperature for 12 h. Water (20 mL) and ethyl acetate (30 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (30 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the residue. The residue was purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 13b (220 mg) in 43% yield.

[0567] MS-ESI calculated value [(1 / 2M)+H] + =673, actual value 673. Phase 2 13b (200 mg, 0.15 mmol) was dissolved in a mixed solvent of anhydrous tetrahydrofuran / anhydrous N,N-dimethylformamide (V テトラヒドロフラン :V N、N-ジメチルホルムアミド The reaction mixture was dissolved in 1:1 HCl (5.4 mL), piperidine (66 mg, 0.78 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was directly purified by silica gel column chromatography (methanol:dichloromethane = 0-100%) to give compound 13c (110 mg) in a 66% yield.

[0568] MS-ESI calculated value [M+H] + =1123.7, actual value 1123.4. Phase 3 13c (110 mg, 0.10 mmol) was dissolved in anhydrous N,N-dimethylformamide (3 mL), 2,4,6-trimethylpyridine (25 mg, 0.21 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (59 mg, 0.16 mmol) were added, and the mixture was stirred at 0 °C for 5 min. Then, 4c (55 mg, 0.19 mmol) was added. After the addition was complete, the mixture was stirred at 0 °C for 1 h. Water (5 mL) and ethyl acetate (10 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC to give compound 13 (54 mg) in 43% yield.

[0569] MS-ESI calculated value [M+H] + =1290.7, actual value 1290.4. 1 H NMR(500MHz,DMSO-d6)δ 8.05(d,J=8.5Hz,1H),7.58-7.52(m,2H),7.36-7.21(m,5H),7.21-7. 10(m,1H),7.02(s,2H),5.14-4.91(m,2H),4.60-4.52(m,1H),4.51-4. 44(m,1H),4.44-4.36(m,1H),4.36-4.30(m,1H),4.28-4.20(m,1H),4.20-4.14(m,1H),4.05-3.90(m,3H),3.85(dd,J=11.0,9.0Hz,1H),3.5 9-3.51(m,1H),3.25-3.20(m,3H),3.21-3.15(m,3H),3.10(s,1H),3. 07-2.90(m,3H),2.88-2.80(m,2H),2.43-2.34(m,1H),2.30-2.20(m,1 H),2.16-1.88(m,4H),1.85-1.63(m,4H),1.62-1.22(m,6H),1.06-0. 96(m,6H),0.89-0.86(m,2H),0.85-0.80(m,10H),0.80-0.70(m,12H).

[0570] Example 1.12

[0571] [ka]

[0572] Phase 1 Under nitrogen gas protection, imidazole (161.60 g, 2373.68 mmol) was dissolved in anhydrous N,N-dimethylformamide (1.25 L), the mixture was cooled to 0±5°C, triisopropylsilyl chloride (458.20 g, 2376.56 mmol) was added dropwise, and the mixture was maintained at 0±5°C while stirring for 30 minutes, and the reaction mixture was added with 14a (250. A solution of 1000g (949.67mmol) of 1000mg of methylsilane in N,N-dimethylformamide (1250mL) was added dropwise. After the addition was complete, the mixture was maintained at 0±5°C and stirred for 18 hours. Imidazole (129.30g, 1899.24mmol) was added and stirred for 10 minutes. Triethylchlorosilane (286.50g, 1900.86mmol) was then added dropwise. After the addition was complete, the mixture was maintained at 0±5°C and stirred for 18 hours. Methanol (250mL) was added dropwise and the mixture was maintained at 0°C and stirred for 1 hour. The mixture was returned to room temperature, diluted with water (5000mL), and the aqueous phase was extracted with methyl t-butyl ether (5000mL x 2). The organic phases were combined. The organic phase was washed with aqueous sodium chloride solution (2500 mL × 2, 5%), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (n-heptane:ethyl acetate = 0 to 100%) to obtain 14b (479.10 g), with a purity of 99.95%, a content of 93.5%, and a discounted content yield of 88%.

[0573] MS-ESI calculated value [M+H] + =534, actual value 534. 1H NMR(400MHz,CDCl3)δ 6.70(s,1H),6.14(d,J=16.4Hz,1H),5.69-5.66(m,1H),5.59(d,J=16.4Hz,1H),5.16(d,J=16.0Hz,1H),4.53(d,J=2 .4Hz,2H),1.87-1.75(m,2H),1.37-1.23(m,3H),1.13(d,J=7.6Hz,18H),0.95-0.87(m,12H),0.67(q,J=7.6Hz,6H).

[0574] Phase 2 Under nitrogen gas protection, 14b (450.0 g, content: 93.5%, 788.13 mmol) was dissolved in toluene (4.5 L), sodium carbonate (25.10 g, 236.81 mmol) and Lawesson's reagent (382.50 g, 945.68 mmol) were added, and the mixture was heated to 110 °C with stirring for 3 h. The reaction mixture was cooled to 50-60 °C, and n-heptane (4500 mL) was added dropwise. After stirring for 1 h, the mixture was filtered. The filter cake was eluted with n-heptane (900 mL). The organic phases were combined, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (n-heptane:ethyl acetate = 0-100%) to give compound 14c (517.0 g), which was used directly in the next step.

[0575] MS-ESI calculated value [M+H] + =550, actual value 550. 1 H NMR(400MHz,CDCl3)δ 7.20(s,1H),6.14(d,J=16.4Hz,1H),5.87(t,J=2.0Hz,1H),5.26(d,J=16.8Hz,1H),4.82(d,J=2.0Hz, 1H), 1.90-1.75 (m, 2H), 1.41-1.27 (m, 3H), 1.18-1.11 (m, 18H), 0.97-0.85 (m, 12H), 0.77-0.66 (m, 6H).

[0576] Phase 3 Under nitrogen gas protection, the crude product 14c (517.0 g) from the previous step was dissolved in anhydrous tetrahydrofuran (4.5 L). Hydrofluoric acid (295.50 g, 48 wt.%, 7092.00 mmol) was added at room temperature. After the addition was complete, the mixture was stirred at room temperature for 36 hours. The reaction mixture was diluted with water (4500 mL). The aqueous phase was extracted with ethyl acetate (4500 mL x 2). The organic phases were combined. The organic phase was washed with saturated brine (2250 mL x 2). The organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (dichloromethane:ethyl acetate = 0-100%) to give the crude product. The crude product was slurried with methyl t-butyl ether (2250 mL) for 1 hour and filtered to give compound 14d (194.60 g). The content was 94% and the two-step yield was 83%.

[0577] MS-ESI calculated value [M+H] + =280, actual value 280. 1 H NMR(400MHz,CDCl3)δ 6.63(s,1H),5.82(d,J=17.2Hz,1H),5.41(d,J=17.2Hz,1H),4.45-4.30(m,2H),2.99-2.92(m,2H),1.80(q,J=7.2Hz,2H),0.78(t,J=7.2Hz,3H).

[0578] Phase 4 Under nitrogen gas protection, add water (8.0 L) and HCl(aq) (8.0 L, 6N) to a 50 L reactor, add 14e (800.00 g, 3196.55 mmol), and heat the mixture to 100 ± 5 °C. Allow the mixture to react for 4 hours with stirring. Cool the mixture to 30 °C and slowly add aqueous ammonia (3500 mL, 25%-28%) dropwise to adjust the pH to 4-5 (control the reaction temperature below 40 °C during the addition). After the addition is complete, stir the mixture for 30 minutes, filter, and dissolve the filter cake in water (2500 mL x 1). Slurry the filter cake in ethanol (8 L) at room temperature for 1 hour, filter, and dissolve the filter cake in ethanol (1000 mL x 1). The filter cake was collected and air dried at 50±5° C. to give 14f (709.00 g), yield: 91%.

[0579] MS-ESI calculated value [M+H] + =209, actual value 209. Stage 5 Tetrahydrofuran (10,200 mL), water (10,200 mL), and 14f (680.00 g, 2779.03 mmol) were added to a 50 L reactor. The mixture was cooled to 0-10°C, followed by the addition of potassium carbonate (653.00 g, 4724.69 mmol). A solution of 9-fluorenylmethyl chloroformate (733.31 g, 2834.60 mmol) in tetrahydrofuran (3,400 mL) was slowly added dropwise (controlling the reaction temperature at 0-10°C during the addition). The mixture was stirred and maintained at 0-10°C for 1 hour. The reaction mixture was extracted with 2-methyltetrahydrofuran (10,200 mL x 2), and the organic phase was washed sequentially with water (10,200 mL x 1) and saturated brine (10,000 mL x 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. Methyl t-butyl ether (6800 mL) was added to the residue, stirred at room temperature for 4 hours, filtered, and the filter cake was eluted with methyl t-butyl ether (680 mL × 1). The filter cake was collected and air-dried at 50 ± 5 °C to give 14 g (1090.00 g), a yield of 91%.

[0580] MS-ESI calculated value [M+H] + =431, actual value 431. 1 H NMR(400MHz,DMSO-d6)δ 7.90(d,J=7.2Hz,2H),7.80-7.73(m,2H),7.53-7.28(m,6H),6.39(d,J=12.8Hz,1H),4. 40-4.15(m,4H),3.03-2.75(m,2H),2.20-2.07(m,1H),2.04-1.87(m,1H),1.98(s,3H).

[0581] Stage 6 Under nitrogen gas protection, 7g (150.00g, 483.47mmol) was added to a three-necked bottle (3L), ethyl acetate (1200mL) was added, and a solution of 14h (111.20g, content: 90%, 531.69mmol) in ethyl acetate (300mL) was added dropwise. After the addition was completed, the mixture was maintained at room temperature and stirred for 2-6 hours. Saturated saline (450mL) was added to the reaction system, and the mixture was stirred at room temperature for 10 minutes. After that, the liquid was separated and the organic phase was kept. The organic phase was washed with saturated saline (450mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the residue. The residue was then purified by silica gel column chromatography ( Purification with ethyl acetate:petroleum ether=0-100% gave compound 14i (134.00 g), yield: 73%.

[0582] MS-ESI calculated value [M+H] + =384, actual value 384. 1 H NMR(400MHz,DMSO-d6)δ 8.13-8.05(m,2H),7.01(s,2H),3.71(d,J=6.0Hz,2H),3.70(d,J=6.0Hz,2H ),3.61-3.52(m,4H),3.50-3.44(m,2H),2.33(t,J=6.4Hz,2H),1.40(s,9H).

[0583] Stage 7 Under nitrogen gas protection, 14i (184.30 g, 480.70 mmol) was added to a three-necked bottle (3 L), toluene (1.84 L) was added, and trifluoroacetic acid (823.27 g, 7220.21 mmol) was added dropwise. After the addition was complete, the mixture was maintained at room temperature and stirred for 4.5 hours. The reaction mixture was concentrated under reduced pressure to obtain a residue. Toluene (600 mL) was added to the residue, and the mixture was continuously concentrated under reduced pressure three times until a solid precipitated. Ethyl acetate (1.7 L) was added to the solid, and the mixture was slurried for 2 hours and then filtered to obtain compound 8g (150.20 g), yield: 96%.

[0584] MS-ESI calculated value [MH]-=326, observed value 326. 1 H NMR(400MHz,DMSO-d6)δ 8.12-8.05(m,2H),7.01(s,2H),3.75(d,J=6.0Hz,2H),3.70(d,J=5.6Hz,2H),3.60-3.52(m,4H),3.50-3.44(m,2H),2.33(t,J=6.4Hz,2H).

[0585] Stage 8 Under nitrogen gas protection, dissolve 8g (20.00g, 61.11mmol) in dichloromethane (200mL) and cool to 0±5°C. Then, add pentafluorophenol (16.90g, 91.82mmol) and react for 10 minutes with stirring. Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (17.60g, 91.82mmol) and react for 4 to 16 hours with stirring while maintaining the mixture at 0±5°C. The reaction mixture was diluted with water (200 mL), stirred for 10 minutes, and then separated. The organic phase was washed with saturated brine (100 mL × 1), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (acetone:dichloromethane = 0 to 100%) to give a crude product. Methyl t-butyl ether (150 mL) was added to the crude product, stirred at room temperature for 1 hour, filtered, and the filter cake was dried at 35 °C to give compound 14j (23.00 g), yield: 76%.

[0586] MS-ESI calculated value [M+H] + =494, actual value 494. 1 H NMR(400MHz,CDCl3)δ 7.12(d,J=5.6Hz,1H),6.96(t,J=5.6Hz,1H),6.70(s,1H),4.42(d,J=6.0Hz,2H),4.03(d,J=6.0H z,1H),3.72(t,J=5.2Hz,2H),3.69(t,J=5.6Hz,2H),3.59(t,J=5.2Hz,2H),2.49(t,J=5.6Hz,2H).

[0587] Stage 9 Under nitrogen gas protection, 8e (7.50 g, 24.25 mmol) was dissolved in tetrahydrofuran (400 mL) and stirred at room temperature for 1 hour. 14j (10.00 g, 20.27 mmol) was added. After the addition was complete, the mixture was stirred at room temperature for 16 hours. Ethyl acetate (400 mL) was added to the reaction mixture, and the mixture was stirred for 1 hour. The mixture was filtered, the filter cake was dried, and the filter cake was diluted with dichloromethane / methanol (v:v = 20:1, 10 The crude product (14.51 g) was obtained by dissolving the crude product in methanol (30 mL), adding ethyl acetate (450 mL) dropwise to the methanol solution, stirring at room temperature for 24 hours, filtering, and drying the filter cake to obtain compound 7i (10.20 g), yield: 81%.

[0588] MS-ESI calculated value [M+Na] + =641, actual value 641. Stage 10 Under nitrogen gas protection, toluene (3200 mL), acetic acid (3200 mL), 14d (320.00 g, 1145.68 mmol), 14g (592.00 g, 1375.21 mmol), and p-toluenesulfonic acid pyridine salt (144.00 g, 573.02 mmol) were sequentially added to a 10 L reaction bottle. The mixture was heated to 110 °C and stirred for 24 h. The reaction mixture was cooled to 30 ± 10 °C and transferred to an 80 L extraction vessel. 2-Methyltetrahydrofuran (6400 mL) and water (6400 mL) were sequentially added. The mixture was stirred for 10 to 20 min, separated, and the upper organic phase was retained. The organic phase was washed successively with water (4800 mL × 3) and saturated aqueous sodium chloride solution (4800 mL × 1), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure (50 ± 5 °C) until almost no liquid was dripping. The residue was evaporated twice with methyl t-butyl ether (640 mL × 2). The obtained crude product 1 was slurried in methyl t-butyl ether (3200 mL) for 2 to 2.5 hours, filtered, and the filter cake was eluted twice with methyl t-ether (640 mL × 2). The filter cake was collected and dried to obtain crude product 2. The crude product 2 was dissolved in N,N-dimethylformamide (6400 mL), and methyl t-butyl ether (25600 mL) was added dropwise. After the addition was completed, the mixture was stirred and crystallized for 16-18 hours at 20-30°C. The mixture was filtered, and the filter cake was slurried with methyl t-butyl ether (3200 mL) for 2-2.5 hours, followed by filtration. The filter cake was eluted twice with methyl t-butyl ether (640 mL x 2). The filter cake was collected and air-dried (50±5°C) to give 14k (490 g), yield: 63%.

[0589] MS-ESI calculated value [M+H] + =674, actual value 674. 1H NMR(400MHz,DMSO-d6)δ 8.16-8.06(m,1H),7.85(d,J=7.2Hz,2H),7.82-7.74(m ,2H),7.74-7.64(m,2H),7.42-7.33(m,2H),7.33-7.23(m,2H),6.76-6.64(m, 2H),5.97-5.81(m,1H),5.58-5.40(m,2H),5.38-5.20(m,2H),4.68-4.52(m,1 H),4.34(s,2H),3.28-3.14(m,1H),3.12-2.96(m,1H),2.32(s,3H),2.2 8-2.18(m,1H),2.18-2.04(m,1H),2.00-1.82(m,2H),0.94-0.82(m,3H).

[0590] Stage 11 Under a nitrogen atmosphere, tetrahydrofuran (19,200 mL) and 14k (480 g, 712.42 mmol) are added sequentially to the reaction vessel, the mixture is cooled to 0-5°C, piperidine (546 g, 6412.21 mmol) is slowly added dropwise to the reaction bottle (controlling the temperature at 0-5°C during the addition), and after the addition is complete, the reaction is maintained at 0-5°C and reacted for 22-24 hours with stirring. Slowly add aqueous hydrochloric acid (1440 mL, 6N) to the reactor (control the temperature at 0-10°C during addition), and continue stirring the mixture for 10-15 minutes. Concentrate under reduced pressure to obtain a residue. Slurry the residue with water (9600 mL) for 2-2.5 hours, filter, and eluate the filter cake with acetonitrile (480 mL x 2). After drying, the filter cake was separated and purified (separation apparatus: Hanbon industrial preparative liquid chromatograph, specification model: DAC150, preparative column packing: W). The solution was adjusted to pH 7-8 with 5% aqueous ammonia, stirred for 30-35 minutes, filtered, and the filter cake was eluted with water (1000 mL). The filter cake was collected and air-dried at 60°C for 24±2 hours to give 1d (97 g), yield: 30%.

[0591] MS-ESI calculated value [M+H] + =452, actual value 452. Stage 12 Under nitrogen gas protection, N,N-dimethylformamide (1200 mL) was added to a reaction bottle, followed by 1d (60.00 g, 132.89 mmol) and trifluoroacetic acid (15.20 g, 133.31 mmol). The mixture was cooled to 0-10 °C and stirred for 10 min. After that, 7i (115.10 g, 186.07 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (51.00 g, 266.04 mmol), and 2,4,6-trimethylpyridine (16.10 g, 132.86 mmol) were added. The reaction mixture was stirred at 0-10 °C for 1 h. Add aqueous hydrochloric acid (1200 mL, 0.05 N) to the reaction bottle, then add water (1800 mL) dropwise. Stir for 10-20 minutes, filter, wash the filter cake with water (300 mL × 2), collect the filter cake, dissolve the filter cake in dichloromethane / methanol (V / V = 30 / 1, 1800 mL), dry the solution over anhydrous sodium sulfate, and filter. Purify the filtrate by silica gel column chromatography (methanol:dichloromethane = 0-100%) to obtain the crude product. Slurry the crude product with methyl t-butyl ether (600 mL) for 1-1.5 hours, filter, and eluate the filter cake with methyl t-butyl ether (300 mL × 2). Collect the filter cake and suction dry on a rotary evaporator for 5 ± 1 hours to obtain compound 7 (90.0 g), yield: 64%.

[0592] MS-ESI calculated value [M+H] + =1052, actual value 1052. Example 1.13

[0593] [ka]

[0594] Phase 1 Under nitrogen gas protection, 15a (438.00 g, 904.0 mmol) was dissolved in ethylene glycol dimethyl ether (4380 mL), water (2630 mL) was added, the reaction mixture was cooled to 0-5°C, diglyceride peptide (238.87 g, 1808.0 mmol) and sodium bicarbonate (151.87 g, 1808.0 mmol) were added sequentially, and the mixture was heated to 20-25°C and stirred for 1 hour. The reaction mixture was cooled to -5-0°C, and dilute hydrochloric acid (0.5 M, 4380 mL) was added. The mixture was stirred for 10 minutes. The aqueous phase was extracted sequentially with 2-methyltetrahydrofuran (8760 mL × 1) and 2-methyltetrahydrofuran (4380 mL × 1), and the organic phases were combined. The organic phase was washed with saturated brine (4380 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was spin-dried to obtain a crude product. The crude product was slurried in acetone (8760 mL) for 16 hours, filtered, and the filter cake was dried to obtain 8a (375.00 g), yield: 83%.

[0595] MS-ESI calculated value [M+H] + =502, actual value 502. Phase 2 8a (375.00 g, 747.69 mmol) was dissolved in N,N-dimethylformamide (4000 mL), and lead tetraacetate (730.11 g, 1646.70 mmol), copper acetate (135.95 g, 748.50 mmol), and acetic acid (98.80 g, 1646.70 mol) were added sequentially to the reaction bottle. The mixture was heated to 45-50°C and reacted for 0.5 hours with stirring. The reaction mixture was added to 2-methyltetrahydrofuran (7500 mL) and water (7500 mL), and the layers were separated. The aqueous phase was extracted with 2-methyltetrahydrofuran (3750 mL × 1). The organic phases were combined and the organic phases were washed successively with saturated aqueous sodium bicarbonate solution (3750 mL × 1), water (3750 mL × 1) and saturated brine (3750 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. After slurried with methyl t-butyl ether (3750 mL) for 16 hours, it was filtered and the filter cake was air-dried for 16 hours to give 8b (345.00 g), yield: 89%.

[0596] MS-ESI calculated value [M-CH3COO] + =456, actual value 456. 1 H NMR(400MHz,DMSO-d6)δ 8.67(t,J=6.8Hz,1H),8.34(t,J=5.6Hz,1H),7.87(d,J=7.6Hz,1H),7.71-7.58(m,3H),7.50-7.10(m,9H),5.18-5.04(m,2H),4.40 -4.00(m,4H),3.78(d,J=5.6Hz,2H),3.83-3.69(m,2H),3.05(dd,J=13.6,4.0Hz,1H),2.80(dd,J=13.2,10.8Hz,1H),1.98(s,3H).

[0597] Phase 3 Under nitrogen gas protection, 8b (345.00 g, 669.17 mmol) was dissolved in tetrahydrofuran (3450 mL), the reaction system was cooled to 0-5°C, and benzyl glycolic acid (244.47 g, 1472.17 mmol) and p-toluenesulfonic acid (17.20 g, 100.37 mmol) were added sequentially to the reaction system. After the addition was completed, the reaction system was heated to 20-25°C and reacted for 1 hour with stirring. Ethyl acetate (3450 mL) and water (3450 mL) were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (3450 mL × 1). The organic phases were combined and washed successively with water (3450 mL × 1) and brine (3450 mL × 1). The organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated to give the crude product, which was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give 8c (178.00 g) in a 42% yield.

[0598] MS-ESI calculated value [M+Na] + =644, actual value 644. 1H NMR(400MHz,DMSO-d6)δ 8.63(t,J=6.8Hz,1H),8.35(t,J=5.6Hz,1H),7.87(d,J=7.6Hz,1H),7.71-7.60(m,3H),7.45-7.13(m,14H),5.13(s,2H),4.67-4. 61(m,2H),4.32-4.24(m,1H),4.22-4.09(m,5H),3.83-3.69(m,2H),3.06(dd,J=13.6,4.0Hz,1H),2.80(dd,J=13.6,10.4Hz,1H).

[0599] Phase 4 Under nitrogen gas protection, 8c (102.00 g, 164.07 mmol) was dissolved in dichloromethane (1020 mL), the reaction mixture was cooled to -5 to 0 °C, and 1,8-diazobispyro[5.4.0]undecane-7-ene (24.97 g, 164.07 mmol) was added. After the addition was complete, the mixture was stirred at 0 to 5 °C for 1 h. The reaction mixture was directly purified by silica gel column chromatography (dichloromethane:isopropanol = 0 to 100%) to give 15b (43.50 g) in a 66% yield.

[0600] MS-ESI calculated value [M+H] + =400, actual value 400. 1 H NMR(400MHz,DMSO-d6)δ 8.64(t,J=6.4Hz,1H),8.26-8.14(m,1H),7.45-7.15(m,10H),5.15(s,2H),4.64(d,J=6.8Hz,2H),4.15(s,2H) ),3.81-3.65(m,2H),3.44(dd,J=8.4,4.4Hz,1H),2.98(dd,J=13.6,4.8Hz,1H),2.59(dd,J=13.6,8.8Hz,1H).

[0601] Stage 5 Under nitrogen gas protection, 15b (43.00 g, 107.65 mmol) was dissolved in a mixture of tetrahydrofuran (430 mL) and water (430 mL). The mixture was cooled to 0-10 °C, wet Pd / C (6.50 g) was added, and the mixture was purged with hydrogen gas three times. The mixture was stirred at 0-10 °C for 24 h under a hydrogen atmosphere. The reaction mixture was filtered, and the filter cake was washed successively with tetrahydrofuran (43 mL) and water (43 mL). The filtrate was concentrated under reduced pressure to remove tetrahydrofuran. The remaining aqueous phase was washed with 2-methyltetrahydrofuran (430 mL × 2) and then lyophilized to give 8e (31.50 g), yield: 95%.

[0602] MS-ESI calculated value [MH]-=308, observed value 308. 1 H NMR(400MHz,DMSO-d6)δ 9.07(s,1H),8.67(s,1H),7.33-7.26(m,2H),7.25-7.18(m,3H),4.68-4.56(m,2H),3.92-3.71( m,4H),3.52(dd,J=16.4,4.0Hz,1H),3.01(dd,J=13.6,6.4Hz,1H),2.79(dd,J=13.2,7.6Hz,1H).

[0603] Stage 6 4a (1000.00 g, 9515.65 mmol) and acetonitrile (10.00 L) were added to a 20 L reaction bottle. tert-Butyldimethylsilyl chloride (1505.92 g, 9991.42 mmol) was added to the reaction bottle. The reaction mixture was cooled to 0-10 °C, and then 1,8-diazobispiro[5.4.0]undecan-7-ene (1376.18 g, 9039.84 mmol) was slowly added. After the addition was complete, the reaction mixture was allowed to return to room temperature and reacted for 40 h with continuous stirring. The reaction mixture was directly filtered, and the resulting filter cake was dissolved in methanol (18.00 L), filtered, and acetonitrile (54.00 L) was added to the filtrate. The mixture was stirred for 2 h. The solid precipitated, filtered, and the filter cake was collected and dried to give 4b (1481.60 g), yield: 71%.

[0604] MS-ESI calculated value [MH]-=218, found value 218. Stage 7 4b (500.00 g, 2279.36 mmol) and tetrahydrofuran (3.75 L) were added to a reaction bottle, the reaction mixture was cooled to 0-5°C, and N-methoxycarbonylmaleimide (395.00 g, content: 85%, 2164.59 mmol) was added. After the addition was complete, the mixture was returned to room temperature and stirred for 16 hours. The reaction mixture was cooled again to 0-5°C, and 10% aqueous sodium bicarbonate solution (3.75 L) was added (controlling the temperature at <20°C during the addition). After the addition was complete, the reaction mixture was warmed to 30°C and stirred for 88 hours. The reaction mixture was cooled to 0-5°C, and 5% aqueous citric acid was added dropwise to adjust the pH to 5-6. Ethyl acetate (3.75 L) was added, and the mixture was separated. The aqueous phase was extracted with ethyl acetate (1.50 L). The organic phases were combined, washed sequentially with water (3.75 L × 1) and saturated brine (3.75 L × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0~100%) to give 15c (350.00 g), content: 88%, yield: 45%.

[0605] MS-ESI calculated value [M+H] + =300, actual value 300. 1 H NMR (400MHz, DMSO-d6) δ 13.35(br s,1H),7.14(s,2H),4.69-4.81(m,1H),4.13-3.97(m,2H),0.77(m,9H),0.00(s,3H),-0.06(s,3H).

[0606] Stage 8 Dichloromethane (1260 mL) and 15c (210.00 g, 701.40 mmol) were added to the reaction bottle, and the reaction mixture was cooled to 0-5°C. N,N'-dicyclohexylcarbamate was added. A solution of 159.30 g (772.56 mmol) in dichloromethane (420 mL) and a solution of tetrafluorophenol (122.46 g (737.40 mmol) in dichloromethane (420 mL) were added sequentially, and the reaction mixture was stirred and reacted for 1 h at 0-5 °C. The reaction mixture was filtered, the filter cake was washed with dichloromethane (1050 mL), and the filtrate was spin-dried to obtain a residue. 2100 mL of methyl t-butyl ether was added to the residue, and the mixture was stirred for 10 min, then filtered. The filter cake was eluted with 630 mL of methyl t-butyl ether. The filtrate was concentrated to obtain a residue. 2100 mL of n-hexane was added to the residue, and the mixture was slurried for 30 min, then filtered. The filter cake was washed with n-hexane (630 mL), filtered, and dried to obtain 15d (246.78 g), yield: 79%.

[0607] MS-ESI calculated value [M+H] + =448, actual value 448. 1 H NMR(400MHz,CD3OD)δ 7.48-7.36(m,1H),6.98(s,2H),5.30(dd,J=9.2,6.0Hz,1H),4.34-4.23(m,2H),0.84(s,9H),0.07(s,3H),0.02(s,3H).

[0608] Stage 9 Add water (1470 mL) and diglycine peptide (79.58 g, 602.91 mmol) to a reaction bottle, start stirring, add ethylene glycol dimethyl ether (2450 mL), cool the reaction mixture to 5-10 °C, add 15d (245.00 g, 547.53 mmol) and sodium bicarbonate (92.09 g, 1096.18 mmol) to the reaction mixture, and react for 40 hours with stirring. The reaction mixture was washed with methyl t-butyl ether (2450 mL × 2), 2-methyltetrahydrofuran (7350 mL) was added to the aqueous phase, and pre-cooled aqueous citric acid solution (3920 mL, 2.5%) was added to adjust the pH to 5-6 (control the temperature at approximately 0°C during the dropwise addition), followed by separation. The aqueous phase was extracted with 2-methyltetrahydrofuran (2450 mL × 1), and the organic phases were combined. The organic phase was washed successively with water (7350 mL × 4) and saturated aqueous sodium chloride solution (3680 mL × 1), and the organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to give a residue, which was slurried in ethyl acetate (2450 mL) for 10 minutes and then filtered. The filter cake was washed with ethyl acetate (735 mL). The filtrate was again concentrated under reduced pressure to give a residue, which was dissolved in ethyl acetate (1230 mL) and added dropwise with n-hexane (6125 mL). A solid precipitated, which was stirred for 16 hours and then filtered. The filter cake was eluted with n-hexane (735 mL) and dried to give 15e (115.40 g), yield: 51%.

[0609] MS-ESI calculated value [M+H] + =414, actual value 414. 1 H NMR(400MHz,DMSO-d6)δ 8.41(t,J=5.6Hz,1H),8.05(t,J=5.6Hz,1H),7.11(s,2H),4.73(dd,J=10.4,5.2Hz,1H),4.11(dd,J=10.4,5.6Hz,1H ),4.00(t,J=10.4Hz,1H),3.83-3.73(m,3H),3.57(dd,J=16.8,5.6Hz,1H),0.76(s,9H),-0.01(s,3H),-0.07(s,3H).

[0610] Stage 10 Dichloromethane (1150 mL) and 15e (115.00 g, 278.11 mmol) were added to a reaction bottle, the reaction system was cooled to -40 to -30 °C, and a solution of N,N'-dicyclohexylcarbodiimide (86.10 g, 417.29 mmol) in dichloromethane (155 mL) and a solution of tetrafluorophenol (46.24 g, 278.44 mmol) in dichloromethane (155 mL) were added sequentially. The reaction was carried out for 5 hours with stirring while maintaining the temperature at -40 to -30 °C. Water (2300 mL) was added to quench the reaction, and the reaction solution was filtered. The insoluble matter was removed by stirring, and the filtrate was separated. The aqueous phase was extracted with dichloromethane (1150 mL). The combined organic phases were washed with saturated brine (1730 mL × 1), dried over anhydrous sodium sulfate, and then filtered. The filtrate was concentrated under reduced pressure to give a residue. The residue was slurried with ethyl acetate (1150 mL) for 30 minutes and then filtered. The filter cake was washed with ethyl acetate (345 mL). The filtrate was collected and concentrated. The residue was slurried with (methyl t-butyl ether:n-hexane = 2:13, 1730 mL) for 3 hours and then filtered. The filter cake was eluted with n-hexane (345 mL). The filter cake was collected and dried to give 15f (146.80 g), yield: 94%.

[0611] MS-ESI calculated value [M+H] + =562, actual value 562. 1 H NMR(400MHz,CDCl3)δ 7.70-7.61(m,1H),7.14-7.06(m,1H),7.06-6.96(m,1H),6.75(s,2H),4.72(dd,J=8.4,6.4Hz,1H),4.50-4.35(m,2 H),4.35-4.24(m,1H),4.16-3.94(m,2H),3.87(dd,J=10.4,6.4Hz,1H),0.87(s,9H),0.085(s,3H),0.078(s,3H).

[0612] Stage 11 Acetonitrile (2628 mL), water (876 mL), and 15f (146.00 g, 260.00 mmol) were added to a reaction bottle, and the reaction mixture was cooled to −5 to 0°C. 8e (96.60 g, 312.30 mmol) and N,N-diisopropylethylamine (33.70 g, 260.65 mmol) were added, and the reaction mixture was maintained at −5 to 0°C while stirring for 18 hours. Methyl t-butyl ether (2920 mL) and water (1460 mL) were added to the reaction mixture, stirred, and then the mixture was separated. The organic phase was extracted with water (1460 mL × 1). The aqueous phases were combined and washed with methyl t-butyl ether (2190 mL × 3). The aqueous phase was collected and a phosphate buffer solution was added (2190 mL, sample ratio: 6.51 g of sodium dihydrogen phosphate and 0.30 g of disodium hydrogen phosphate dissolved in 100.0 mL of water). The mixture was stirred for 10 to 20 minutes and extracted with 2-methyltetrahydrofuran (2190 mL × 3). The organic phases were combined and washed with saturated brine (2190 mL × 2). The organic phase was dried over anhydrous sodium sulfate and then filtered. The filtrate was concentrated under reduced pressure, and the residue was separated and purified by HPLC (preparative apparatus: Hanbon industrial preparative liquid chromatograph, specification model: DAC150, preparative column packing: Welch Xtimate C18), add about half the volume of the mixed solvent (ethyl acetate: 2-methyltetrahydrofuran = 1:1) to the prepared solution, stir, separate, extract the aqueous phase with about half the volume of ethyl acetate, combine the organic phases twice, wash the organic phase with about half the volume of deionized water three times, concentrate under reduced pressure, and freeze-dry the residue to obtain 15 g (83.54 g), yield: 46%.

[0613] MS-ESI calculated value [M+Na] + =727, actual value 727. Stage 12 Acetonitrile (664 mL), HO (996 mL), and 15g (83.00 g, 117.8 mmol) were added sequentially to the reaction bottle, the reaction mixture was cooled to 5-10 °C, formic acid (65.11 g, 1414.5 mmol) was added, and the reaction mixture was stirred and reacted for 40 hours at 5-10 °C. Methyl t-butyl ether (830 mL) was added to the reaction mixture, and the layers were separated. The organic phase was extracted with water (420 mL × 2), the aqueous phases were combined, and the aqueous phase was lyophilized to obtain the crude product. Dichloromethane (1660 mL) was added to the crude product, which was concentrated under reduced pressure. This procedure was repeated five times (to remove residual formic acid), and then the crude product was dissolved in water (830 mL). The aqueous solution was lyophilized again to obtain 15g (65.30 g), yield: 94%.

[0614] MS-ESI calculated value [M+H] + =591, actual value 591. 1 H NMR(400MHz,DMSO-d6)δ 8.54(t,J=6.4Hz, 1H),8.37-8.28(m,1H),8.12(d,J=7.6Hz,1H),8.02(t,J=6.4Hz,1H),7.29-7.15(m,5H),7.05(s,2H),4.65-4.57(m,3H),4.55-4.45(m,1H) ,4.02-3.94(m,3H),3.92-3.82(m,1H),3.80-3.68(m,4H),3.65-3.57(m,2H),3.04(dd,J=13.6,4.4Hz,1H),2.78(dd,J=14.0,10.0Hz,1H).

[0615] Stage 13 Under nitrogen gas protection, add a solution of 1d (120.00 g, 265.77 mmol) in N,N-dimethylformamide (2400 mL) to the reactor, cool the mixture to -10 to -5°C, add 15h (157.20 g, 266.19 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (151.20 g, 397.64 mmol) to the reactor, and slowly add 2,4,6-trimethylpyridine (64.80 g, 534.74 mmol) dropwise to the reactor (maintain the temperature at -10 to -5°C during the addition). After the addition is complete, the reaction mixture is stirred at -10 to -5°C for 4 hours. Slowly add 2% aqueous citric acid (7200 mL) to the reactor, stirring for 10-15 minutes after addition, then filter. Wash the filter cake with water (1200 mL x 3) and dry. The resulting crude product is purified by reverse-phase chromatography (separation apparatus: Hanbon industrial preparative liquid chromatograph, model: DAC150, preparative column packing: Kromasil 100-10-C18(W)). The solution is processed in batches (25.0 ± 5.0 kg per batch). Each batch of solution is transferred to a 50 L reactor, stirring is started, and acetonitrile is slowly added to clarify the solution. Then ethyl acetate (10,000 mL) is added, stirring for 5-10 minutes, and the solution is allowed to stand for 5-10 minutes to separate the layers. The organic phase was washed with water (10,000 mL x 3), and the combined aqueous phase was back-extracted with ethyl acetate (10,000 mL). The two organic phases were combined and washed with water (10,000 mL x 2). The organic phase was concentrated on a rotary evaporator until no dripping remained. The material in the rotary evaporator bottle was transferred to a 10 L Buchner funnel and suction filtered. The filter cake was then suction filtered until no dripping remained. The rotary evaporator bottle was then rinsed with distilled water (1,200 mL x 2), and the filter cake was eluted. The wet product from the above batches was then combined, added to water (2,400 mL), stirred at 0-10 °C for 2-3 hours, and filtered. The filter cake was eluted with water (480 mL). The filter cake was collected and dried in a vacuum oven (30-35 °C) for 40 hours to obtain compound 4 (152.00 g) in a 56% yield.

[0616] MS-ESI calculation value [M+H] + =1024, measured value 1024. 1 H NMR(400MHz,DMSO-d6)δ 8.70-8.62(m,1H),8.56(d,J=8.8Hz,1H),8.35-8.24(m,2H),8.09(d,J=8.0Hz,1 H),8.04-7.95(m,1H),7.76(s,1H),7.69(d,J=8.8Hz,1H),7.28-7.13(m,5H),7. 05(s,2H),6.69(s,1H),5.90(d,J=16.4Hz,1H),5.72-5.62(m,1H),5.48(d,J=16 .8Hz,1H),5.42(d,J=20.0Hz,1H),5.17(d,J=19.6Hz,1H),5.06(t,J=5.6Hz,1H), 4.68(d,J=7.2Hz,2H),4.60(dd,J=9.6,6.0Hz,1H),4.52-4.43(m,1H),4.18-4.0 4(m,2H),4.01-3.92(m,1H),3.90-3.81(m,1H),3.79-3.68(m,4H),3.65-3.55(m, 2H),3.28-3.07(m,2H),3.01(dd,J=13.6,4.0Hz,1H),2.73(dd,J=13.6,9.6Hz,1 H),2.35(s,3H),2.28-2.12(m,2H),1.88(q,J=7.2Hz,2H),0.85(t,J=7.2Hz,3H).

[0617] Example 1.14

[0618]

change

[0619] Stage 1 Dissolve 8a (200.00 g, 399.12 mmol) in N,N-dimethylformamide (600 mL) and reserve. Add N,N-dimethylformamide (400 mL), acetic acid (119.80 g, 1996.67 mmol), and lead tetraacetate (260.00 g, 598.66 mmol) to a reaction bottle in that order, then slowly add the N,N-dimethylformamide solution of 8a to the reaction bottle. After the addition is complete, heat the mixture to 40°C and react for 4 hours with stirring. Dichloromethane (4000 mL) and water (4000 mL) were added to the reaction mixture, which was stirred for 30 minutes. The insoluble matter was removed by filtration. The filtrate was separated into layers, and the aqueous phase was extracted with dichloromethane (1000 mL × 1). The organic phases were combined and washed with water (2000 mL × 3). The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was slurried with methyl t-butyl ether (1000 mL) for 1 hour, then filtered. The filter cake was collected and dried to obtain 8b (179.63 g), yield: 87%.

[0620] MS-ESI calculated value [M-CH3COO] + =456, actual value 456. Phase 2 Under nitrogen gas protection, 8b (300.00 g, 582.29 mmol) and tetrahydrofuran (1200 mL) were added to a reaction bottle, and the reaction system was cooled to 0-5°C. After that, benzyl glycolic acid (106.39 g, 640.50 mmol) was added (the remaining benzyl glycolic acid was washed with 300 mL of tetrahydrofuran and then added to the reaction bottle). Then, a solution of lithium hydroxide (16.70 g, 698.74 mmol) in water (300 mL) was added dropwise. After the addition was completed, the reaction system was maintained at 0-5°C and reacted with stirring for 2 hours. The reaction mixture was diluted with water (3000 mL) and dichloromethane (3000 mL), stirred, and then separated. The organic phase was washed with brine (4000 mL × 1) and concentrated under reduced pressure to approximately 450 g. The resulting crude product solution was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 0-100%) to give 8c (269.20 g), yield: 74%.

[0621] MS-ESI calculated value [M+Na]+ =644, actual value 644. Phase 3 Under nitrogen gas protection, 8c (264.00 g, 424.98 mmol) was dissolved in dichloromethane (26440 mL), the reaction mixture was cooled to -5 to 0°C, and 1,8-diazobispyro[5.4.0]undecane-7-ene (64.60 g, 424.72 mmol) was added. After the addition was complete, the mixture was stirred at 0 to 5°C for 2 hours. The reaction mixture was then directly transferred onto silica gel. The resulting mixture was purified by column chromatography (dichloromethane:isopropanol = 0 to 100%) to give 15b (111.00 g), yield: 66%.

[0622] MS-ESI calculated value [M+H] + =400, actual value 400. Phase 4 15b (108.00 g, 270.60 mmol) and water (2160 mL) were added to a reaction bottle, followed by the addition of wet Pd / C (10.80 g). The mixture was purged with hydrogen gas three times and reacted under a hydrogen atmosphere with stirring for 3 hours. The reaction mixture was filtered to remove insoluble matter, and the aqueous phase was directly lyophilized to give 8e (82.16 g), yield: 98%.

[0623] MS-ESI calculated value [MH]-=308, observed value 308. Example 2 Method for preparing antibody-drug conjugates Anti-IGF-1R antibody sequences were used to prepare the antibodies, which have the CDR regions (Kabat definitions) as shown below.

[0624] CDR1:SFVMA (SEQ ID No. 1) CDR2:AISGSGSRARYADSVKG(SEQ ID No.2) CDR3:NPRRATPDLTQYAY(SEQ ID No.3) Heavy chain variable region: EVQLVESGGGLVQPGGSLRLSCAASGRTFSSFVMAWFRQAPGKGLEFVSAISGSGSRARYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPRRATPDLTQYAYWGQGTLVTVSS(SEQ ID No.4) The full length sequence is as follows:

[0625] [Table 2]

[0626] The antibody is dialyzed against 50 mM PB buffer to obtain an antibody intermediate. A suitable amount of the antibody intermediate is taken and diluted with 10 mM tris(2-carboxyethyl)phosphine hydrochloride (TCEP) mother solution and 1 0 mM diethylenetriaminepentaacetic acid (DTPA) mother solution was added sequentially, followed by 50 mM PB buffer, resulting in a final antibody concentration of 20 mg / mL, a TCEP to antibody molar ratio of 4, and a final DTPA concentration of 1 mM. After thorough mixing, the mixture was placed in an incubator mixer at 25±2°C and a rotation speed of 400 rpm for 2 hours for reduction. After reduction was complete, an appropriate amount of 5 mM linker-drug mother solution was added sequentially to each reaction system in an ice-water bath, resulting in a linker-drug to antibody molar ratio of 4.5. DMSO was added until the final volume of DMSO in the coupling reaction was 20%. After thorough mixing, the mixture was placed in an incubator mixer at 25±2°C and a rotation speed of 400 rpm for 1 hour for coupling. After coupling is complete, the ADC sample is dialyzed using ultrafiltration centrifuge tubes against dialysate (10 mM His / His-HCl, pH 6.0±0.2) to obtain an ADC stock solution, which is stored in aliquots at -80°C.

[0627] The purity and average drug-to-antibody coupling ratio (DAR) of the ADC molecular size variants were determined using high-performance size-exclusion liquid chromatography (SEC-HPLC) and high-performance liquid hydrophobic chromatography (HIC-HPLC), respectively, and the main parameters are shown in Tables 1 and 2. The purity and drug-to-antibody coupling ratio results of the prepared ADCs are shown in Table 3. The detection results indicate that the prepared ADCs all have high purity and a uniform distribution of drug-to-antibody coupling ratios.

[0628] [Table 3]

[0629] [Table 4]

[0630] [Table 5]

[0631] Example 3 ADC plasma stability study The linker-payload of the present application is obtained by reference to the method in Example 1 of the present application, and the antibody-drug conjugate (ADC) of the present application is obtained by reference to the method in Example 2 of the present application.

[0632] An appropriate amount of test ADC stock solution was added to 8 mL of anticoagulated human plasma to achieve an ADC concentration of 200 μg / mL. After incubation in a biochemical incubator at 37°C for 0, 24, 72, or 168 hours, the sample was collected and purified by protein A chromatography. The purified ADC sample was concentrated by ultrafiltration centrifugation. After centrifugation was complete, 5 μL of 5 M tris(2-carboxyethyl)phosphine hydrochloride (TCEP) mother solution and an appropriate amount of ultrapure water were added to bring the total sample volume to approximately 200 μL. The sample was then placed in a dry incubator at 56°C for 40 minutes, removed, and subjected to another ultrafiltration centrifugation (12,000 rpm, 15 minutes). After centrifugation was complete, 100 μL of ultrapure water was added and mixed uniformly before RP-MS analysis. The mass spectrometer was set in positive mode, and the mobile phase was A: 0.1% formic acid water, B: 0.1% formic acid acetonitrile, the column temperature was 40°C, and the flow rate was 0.4 mL / min.

[0633] The mass numbers of small molecule drugs coupled with different numbers of drugs were analyzed according to the molecular weight. The average coupling rate (Drug-to-antibody ratio, DAR) was calculated according to the ratio of each peak area and the number of drugs coupled. The plasma stability of the ADC was investigated by measuring the change in DAR value after different incubation times. The detection results are shown in Table 1. 4. Here, the formula for calculating the average coupling rate is as follows:

[0634] DAR=(0*DAR0%+1*DAR1%+2*DAR2%) / (DAR0%+DAR1%+DAR2%)×2 The experimental results show that all of the ADCs prepared in this application exhibit good plasma stability.

[0635] [Table 6]

[0636] Example 4 Detection of in vitro growth inhibition of human tumor cells The linker-payload of the present application is obtained by reference to the method in Example 1 of the present application, and the ADC of the present application is obtained by reference to the method in Example 2 of the present application.

[0637] Human tumor cells in the logarithmic growth phase were harvested, digested, resuspended in fresh complete culture medium, and adjusted to an appropriate concentration. They were then added to a 96-well cell culture plate, which was then placed in a 37°C, 5% CO2 incubator and cultured overnight. The next day, different concentrations of the test ADC samples (maximum final concentration 1 μM, 1:4 or 1:8 gradient dilutions) or buffer control were added to the corresponding wells of the cell culture plate. After continued culture in a carbon dioxide incubator for 168 hours, the test plate was equilibrated to room temperature and subjected to CellTiter Glo assay. Luminescence readings were detected using a kit (Promega, G7558) and a multi-function microplate reader. The cell inhibition rate was calculated according to the following formula: Inhibition rate (%) = (1 - (RLU) ADC -RLU 空白 ) / (RLU 緩衝液 -RLU 空白 )) × 100%. The drug inhibition curve was plotted using Graphpad Prism software and the EC 50 The EC values ​​of each ADC in various human tumor cells were fitted. 50 The values ​​are as shown in Table 5.

[0638] The ADCs prepared in this application exhibit excellent tumor cell proliferation inhibitory activity in vitro.

[0639] [Table 7]

[0640] Example 5 In vivo antitumor effect measurement The linker-payload of the present application is obtained by reference to the method in Example 1 of the present application, and the ADC of the present application is obtained by reference to the method in Example 2 of the present application.

[0641] Human tumor cells in the logarithmic growth phase were harvested, counted, and adjusted to the appropriate concentration. They were then resuspended on ice in a 1:1 mixture of serum-free medium and Matrigel (Corning 356234) and subcutaneously inoculated into BALB / c nude mice at 200 μL per mouse. After tumors had grown to a measurable size, the long and short diameters of each tumor were measured using calipers, and the tumor volume was calculated according to the following formula: V = (a × b 2 ) / 2, where a represents the long diameter of the tumor and b represents the short diameter of the tumor.

[0642] Average tumor volume is 100-200mm 3 When the tumor volume reaches 100%, the animals are randomly divided into groups according to tumor volume. According to the planned administration scheme, tumor-bearing nude mice are injected with a vehicle control (saline) or different doses of ADC (3 mg / kg or 10 mg / kg) via the tail vein. The major and minor diameters of the tumor are measured twice a week, and the animal weights are recorded. The tumor volume of each group is calculated, and the tumor growth inhibition rate (TGI) is calculated according to the following formula: TGI = 100% × [1 - (TV tT -TV 0T ) / (TV tC -TV 0C )]. Here, TV tT represents the tumor volume of the administration group on the day of measurement, and TV 0T represents the tumor volume of the treatment group at the time of grouping, and TV tC represents the tumor volume in the vehicle control group on the day of measurement, and TV 0C represents the tumor volume of the vehicle control group at the time of grouping.

[0643] The ADCs prepared in this application exhibit excellent in vivo antitumor activity. Example 6 In vivo pharmacokinetic detection in rats Healthy adult Sprague Dawley rats aged 6 to 8 weeks were prepared, and the ADC was injected into the tail vein for approximately 1 minute ± 10 seconds. The administration volume was 5 mL / kg, and the administration concentration was 20 mg / kg. Blood samples were collected 0.083 hours, 1 hour, 2 hours, 8 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 120 hours, 144 hours, and 168 hours after the completion of administration. Serum is separated by centrifugation within ~120 minutes. Total antibody (Tab) and ADC concentrations in the blood samples are detected by conventional ELISA methods.

[0644] The total antibody detection method is briefly as follows: the antibody is coated overnight with Trop2-His at 4°C, blocked with 5% nonfat dry milk at 37°C for 2 hours, and then a standard curve and quality control points are added and incubated for 2 hours. The detection range of the standard curve is 64ng / mL to 0.5ng / mL, starting from 64ng / mL, and diluted two-fold. The quality control points are set to 60ng / mL, 6ng / mL, and 0.6ng / mL. The recovery rate of the quality control points is 80% to 1. The secondary antibody should be 20%, then a goat anti-human K light chain peroxidase antibody (manufacturer: Sigma, catalog number: A7164-1ML) is added at a dilution of 1:8000. The secondary antibody is incubated for 1 hour, washed 8 times with PBST, then TMB is added to develop the color, and the color is stopped with 0.1M sulfuric acid. The plate is read on an OD450 microplate reader, and the blood concentration at different time points is calculated using the microplate reader analysis software SoftMaxPro.

[0645] The ADC detection method is briefly described as follows: the sample is coated overnight with Trop2-His at 4°C, blocked with 5% nonfat dry milk at 37°C for 2 hours, and then a standard curve and quality control points are added and incubated for 2 hours. The detection range of the standard curve is 64ng / mL to 0.5ng / mL, starting from 64ng / mL, and diluted two-fold. The quality control points are set to 60ng / mL, 6ng / mL, and 0.6ng / mL. The recovery rate of the quality control points is 80% to 1. The secondary antibody should be 20%, then a goat anti-human K light chain peroxidase antibody (manufacturer: Sigma, catalog number: A7164-1ML) is added at a dilution of 1:8000. The secondary antibody is incubated for 1 hour, washed 8 times with PBST, then TMB is added to develop the color, and the color is stopped with 0.1M sulfuric acid. The plate is read on an OD450 microplate reader, and the blood concentration at different time points is calculated using the microplate reader analysis software SoftMaxPro.

[0646] The change curve of drug concentration in blood was plotted using ELISA, and the ADC concentration was basically consistent with the total antibody concentration, with extremely low shedding and very stable in blood.

[0647] Example 7 Cell proliferation inhibition experiment KPL-4 tumor cells in the logarithmic growth phase were harvested, resuspended in fresh RPMI 1640 medium, counted, and the cell suspension was diluted to 2 x 10 4The cell suspension was then inoculated into a 96-well cell culture plate at 100 μL per well and placed in a carbon dioxide incubator (37°C, 5% CO2) for overnight incubation. The next day, one of the 96-well plates containing the inoculated cells was removed and equilibrated to room temperature. 100 μL of CellTiter-Glo reagent (Promega, USA), which had been pre-equilibrated to room temperature and mixed evenly, was added to each well of the test plate. After 30 minutes of incubation in the dark, the luminescence value (recorded as the G0 value) was read using a microplate reader. Another parallel plate was prepared, and different concentrations of the test compound or DMSO (final concentration 0.5%) were added to the corresponding wells of the test plate. After 72 hours of incubation in a carbon dioxide incubator, the test plate was equilibrated to room temperature and the cell activity was detected using CellTiter-Glo reagent, which was recorded as the G3 value.

[0648] The cell proliferation rate was calculated according to the following formula: Cell proliferation rate (%) = (mean value of test compound well G3 - mean value of G0) / (mean value of DMSO control well G3 - mean value of G0) x 100. The inhibition curves were fitted and analyzed using Graphpad Prism software. G.I. 50 Calculate the value.

[0649] The compounds prepared in this application exhibit excellent in vivo antitumor activity. Example 8 Bystander lethal activity experiment BxPC3 (human pancreatic cancer cells, ATCC, CRL-1687) and MiaPaCa2 (human pancreatic cancer cells, biocytogen, B-HCL-014) cells were cultured in RPMI1640 + 10% FBS and DMEM / high glucose + 10% FBS, respectively. The cells were digested with trypsin, neutralized with fresh medium, centrifuged at 1000 rpm for 3 minutes, the supernatant was discarded, and the cells were resuspended in RPMI1640 + 10% FBS. After cell counting, the density of BxPC3 cells was determined to be 6 x 10. 4 Adjust the cell density to 1.5 x 10 cells / mL. 4Adjust the concentration to 100 cells / mL. Add 500 μL of BxPC3 cells and 500 μL of MiaPaCa2-luc cells to each well of plate 1 of a 12-well plate. Add 500 μL of MiaPaCa2-luc cells and 500 μL of RPMI1640 medium containing 10% FBS serum to plate 2 of a 12-well plate. Incubate at 37°C in 5% carbon dioxide for 24 hours.

[0650] Prepare the ADC samples at a 40x intermediate concentration (0.2 μM). Add 25 μL of each sample to the corresponding well of a 12-well plate. This will serve as the solvent control. Incubate at 37°C with 5% carbon dioxide for 6 days. Digest the cells in the 12-well plate with trypsin, neutralize with fresh medium, centrifuge at 1000 rpm for 3 minutes, discard the supernatant, resuspend in 1 mL of FACS buffer (PBS + 2.5% FBS), take 20 μL of cells, add 20 μL of trypan blue, and count. Centrifuge the cells in plate 1 at 1000 rpm for 3 minutes, discard the supernatant, resuspend in 100 μL of FACS buffer, add 2 μL of monoclonal antibody, and incubate on ice for 30 minutes. Centrifuge at 2000 rpm for 1 minute at 4°C, discard the supernatant, and resuspend the cells in 150 μL of FACS buffer. Detection was performed using BD FACSVerse. Data were analyzed using Flowjo 7.6.

[0651] As shown in the results, the ADC disclosed in the present invention has a clear bystander killing effect, and the ADC kills negative MiaPaCa2 cells expressing target markers. However, after BxPC3 cells expressing target markers are mixed with negative MiaPaCa2 cells, the ADC also has a killing effect on negative MiaPaCa2 cells expressing target markers.

[0652] All documents mentioned in this application are incorporated by reference in this application as if each document were incorporated by reference individually. Furthermore, after reading the above teachings of the present invention, those skilled in the art will be able to make various changes or modifications to the present invention, and these equivalents will also fall within the scope defined by the appended claims of this application.

Claims

1. A ligand conjugate, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, wherein the ligand conjugate comprises the structure shown in Formula I: 【Chemistry 1】 where L is an optionally substituted connector that is attached to any O, S, or N atom in the P structure; Ab is a ligand, a is a number greater than 0, a is a decimal number or an integer, preferably a is 1 to 16 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16), more preferably a is 2 to 8; P is a group formed by dehydrogenation of the structure shown in formula (II) below: 【Chemistry 2】 where n is 0 or 1, X is N or CR ０ is selected from the group consisting of R ０ H, D, halogen, C １ -C ８ Alkyl group, C １ -C ８ Alkoxy groups, OH, NH ２ , N ３ Or NO ２ is selected from the group consisting of R １ represents a hydrogen atom, a deuterium atom, a halogen, or C １ -C ８ Alkyl group, C １ -C ８ Alkoxy group, C １ -C ８ Halogenated alkyl group, C １ -C ８ Halogenated alkoxy groups, N ３ , NO ２ , N.H. ２ , NH—OH, —NR′R″, —COOR′, —CONR′R″, —NHR′″NR′R″, wherein R′, R″, and R′″ are each independently selected from hydrogen, deuterium, an alkyl group, an aryl group, an arylalkyl group, an acyl group, an alkoxycarbonyl group, and an aryloxycarbonyl group; R ２ , R ３ , R ４ , R ５ and R ６ are each independently a hydrogen atom, a deuterium atom, a halogen, a hydroxy group, a cyano group, or NH ２ , NO ２ , substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ Alkoxy group, substituted or unsubstituted C １ -C ８ alkylthio group, substituted or unsubstituted C １ -C ８ Deuterated alkyl groups, -(CH ２ ) ｍ Bird (C １ -C ４ alkyl)silyl group, -(CH ２ ) ｍ (C ３ -C ８ cycloalkyl), -(CH ２ ) ｍ (3- to 12-membered heterocyclic group), —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ S (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ S(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ S (O) ２ (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ NH (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ NHC(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ OC(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ C(O)(CH ２ ) ｐ R ７ , —CH═N(OtBu), wherein m and p are each independently 0, 1, 2, 3, or 4; Or, R ２ and R ３ together with the carbon atom to which they are attached, form a substituted or unsubstituted C ５ -C ８ forming a carbocyclic ring or a substituted or unsubstituted 5- to 12-membered heterocyclic group, Or, R ３ and R ４ , or R ４ and R ５ together with the carbon atoms to which they are attached, form a saturated or unsaturated 5- to 12-membered carbocyclic ring which is unsubstituted or substituted by one or more Re, unsubstituted or substituted by one or more R ｅ saturated or unsaturated 5- to 12-membered alkyl group substituted by heterocycles, and R ｅ represents a hydrogen atom, a deuterium atom, a halogen atom, a nitrile group, a nitro group, a hydroxy group, an amino group, C １ -C ６ Alkyl-NH-, (C １ -C ６ alkyl) ２ N-, C １ -C ６ Alkyl group, C ２ -C ６ Alkenyl group, C ２ -C ６ Alkynyl group, C １ -C ６ Alkoxy group, allyl group, benzyl group, C ６ -C １２ Aryl group, C １ -C ６ Alkoxy-C １ -C ６ Alkyl group, C １ -C ６ Alkoxy-carbonyl group, phenoxycarbonyl group, C ２ -C ６ Alkynyl-carbonyl group, C ２ -C ６ Alkenyl-carbonyl group, C ３ -C ６ Cycloalkyl-carbonyl group, C １ -C ６ Alkyl-sulfonyl group, phenyl group, 5- to 7-membered heteroaryl group, C ３ -C ８ a cycloalkyl group, a 3- to 12-membered heterocyclic group, —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ S (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ S(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ S (O) ２ (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ NH (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ NHC(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ OC(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ C(O)(CH ２ ) ｐ R ７ wherein m and p are each independently 0, 1, 2, 3, or 4, preferably 0, 1, or 2; R ７ represents a hydrogen atom, a deuterium atom, a halogen atom, a substituted or unsubstituted C １ -C ８ Alkyl group, C １ -C ８ Halogenated alkyl group, C １ -C ８ Deuterated alkyl groups, substituted or unsubstituted C １ -C ８ an alkoxy group, a hydroxy group, an amino group, a cyano group, a nitro group, a mercapto group, a substituted or unsubstituted C １ -C ８ alkylene-OH, substituted or unsubstituted C １ -C ８ Alkylene-NH ２ , S.O. ２ Me, —OC(O) (substituted or unsubstituted C １ -C ４ alkyl), —C(O)(substituted or unsubstituted C １ -C ４ alkyl), a substituted or unsubstituted phenyl group, a substituted or unsubstituted 5- to 7-membered heteroaryl group, a substituted or unsubstituted C ３ -C ８ selected from the group consisting of cycloalkyl groups, substituted or unsubstituted 3- to 12-membered heterocyclic groups; Unless otherwise specified, each of the above "substituted" groups means that one or more hydrogen atoms on the group have been replaced with a hydrogen atom, a deuterium atom, a halogen atom, a nitrile group, a nitro group, a hydroxy group, an amino group, a C １ -C ６ Alkyl-NH-, (C １ -C ６ alkyl) ２ N-, C １ -C ６ Alkyl group, C ２ -C ６ Alkenyl group, C ２ -C ６ Alkynyl group, C １ -C ６ Alkoxy group, halogenated C １ -C ６ Alkyl group, halogenated C ２ -C ６ Alkenyl group, halogenated C ２ -C ６ Alkynyl group, halogenated C １ -C ６ Alkoxy group, allyl group, benzyl group, C ６ -C １２ Aryl group, C １ -C ６ Alkoxy-C １ -C ６ Alkyl group, C １ -C ６ Alkoxy-carbonyl group, phenoxycarbonyl group, C ２ -C ６ Alkynyl-carbonyl group, C ２ -C ６ Alkenyl-carbonyl group, C ３ -C ６ Cycloalkyl-carbonyl group, C １ -C ６ Alkyl-sulfonyl group, phenyl group, 5- to 7-membered heteroaryl group, C ３ -C ８ the ligand conjugate, or a tautomer, meso isomer, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, characterized in that:

2. The L has a structure as shown in the following formula: L １ -8 ２ -8 ３ -8 ４ -8 ５ ; Here, the L １ is an arbitrarily substituted 【Transformation 3】 and R ｄ is H, C １ -C ６ Alkyl group, C １ -C ６ Deuterated alkyl groups, C ３ -C ８ Cycloalkyl group or C ３ -C ８ is a deuterated cycloalkyl group, Said L ２ is an optionally substituted —(CHR) ｍ１ -X １ - (CH ２ CH ２ O) ｎ３ -(CHR) ｍ２ -C(O)-, optionally substituted -(CHR) ｍ１ -X １ -X ２ - (CH ２ CH ２ O) ｎ３ -(CHR) ｍ２ -C(O)-, optionally substituted -X １ -(CHROCHR) ｍ２ -C(O)-, optionally substituted -(CHR) ｐ 1-C(O)-, optionally substituted -(CHR) ｍ１ -X １ -(CHR) ｍ２ -C(O)-, optionally substituted -(CHR) ｍ１ -X １ -(CHR) ｎ３ -X ２ -(CHR) ｍ２ -C(O)-, optionally substituted -X １ -(CHR) ｍ１ -X ２ -(CHR) ｍ２ -C(O)-, optionally substituted -(CH ２ CH ２ O) ｎ３ is a group selected from the group consisting of —C(O)—, X １ and X ２ are each independently —O—, —C(O)—, —C(O)—NR—, an optionally substituted C ６ -C １０ aryl groups, optionally substituted 5- to 9-membered heteroaryl groups, optionally substituted 3- to 8-membered heteroalicyclic groups, and optionally substituted C ３ -C ６ alicyclic groups; Here, each R is independently H, D, (CH ２ ) ｎ４ OH, (CH ２ ) ｎ４ NH ２ , (CH ２ O) ｎ４ (CH ２ CH ２ O) ｎ５ H, (CH ２ O) ｎ４ (CH ２ CH ２ O) ｎ５ CH ３ , (CH ２ ) ｎ４ OCH ３ , (CH ２ CH ２ O) ｎ５ CH ３ , C.H. ２ C(O)NH(CH ２ O) ｎ４ (CH ２ CH ２ O) ｎ５ H, CH ２ C(O)NH(CH ２ O) ｎ４ (CH ２ CH ２ O) ｎ５ CH ３ is selected from the group consisting of wherein m1, m2, n3, n4, and n5 are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12; and p1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, or 8; Said L ３ is a peptide residue, and said L ３ teeth, 【Chemistry 4】 , C.H. ２ C(O)R ｃ and wherein R ｃ teeth, 【Transformation 5】 wherein n1 and n2 are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; Said L ４ is an optionally substituted -L ４ａ - (NR ｂ ) ｎ６ -R １２ -L ４ｂ -, where L ４ａ is not present or L ４ａ is an arbitrarily substituted 【Transformation 6】 where n6 is 0 or 1, and R １２ is a chemical bond, CH ２ , or CD ２ and L ４ｂ is not present or L ４ｂ is an arbitrarily substituted 【Transformation 7】 where R ａ and R ｂ are each independently hydrogen, optionally substituted C １ -C ４ alkyl groups and optionally substituted C １ -C ４ deuterated alkyl groups; Said L ５ is absent or optionally substituted 【Transformation 8】 wherein Y is selected from the group consisting of O, S, or NH; v is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; and R １０ and R １１ are each independently hydrogen, deuterium, or optionally substituted C １ -C ４ alkyl groups, optionally substituted C １ -C ４ Halogenated alkyl groups, optionally substituted C ３ -C ６ Cycloalkyl groups and optionally substituted C ４ -C ８ cycloalkyl groups; alkyl groups; or R １０ and R １１ together with the atoms to which they are attached form an optionally substituted 3- to 6-membered cycloalkyl group, and R １０ and R １１ are each independently a hydrogen atom, a deuterium atom, a halogen atom, or an optionally substituted C １ -C ８ alkyl groups, optionally substituted C １ -C ８ Halogenated alkyl groups and optionally substituted C １ -C ８ deuterated alkyl groups 2. The ligand conjugate of claim 1, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

3. Said L １ teeth, 【Chemistry 9】 characterized in that 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

4. X １ and X ２ are each independently —O—, —C(O)—, —C(O)—NR—, an optionally substituted phenyl group, an optionally substituted pyridyl group, an optionally substituted C ３ -C ６ Cycloalkyl groups, optionally substituted 【Chemistry 10】 or optionally substituted 【Chemistry 11】 characterized in that the compound is selected from the group consisting of 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

5. Said L ２ is an optionally substituted —(CH ２ ) ｍ１ -X １ - (CH ２ CH ２ O) ｎ３ - (CH ２ ) ｍ２ -C(O)-, where X １ is —C(O)—NH—, and preferably, m1 and m2 are each independently selected from 1, 2, or 3, and n3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

6. Said L ２ is an optionally substituted —(CHR) ｍ１ -X １ -X ２ - (CH ２ CH ２ O) ｎ３ -(CHR) ｍ２ -C(O)-, where X １ is an arbitrarily substituted 【Chemistry 12】 , or optionally substituted 【Chemistry 13】 and X ２ is —C(O)—NR—, and preferably, m1 and m2 are each independently selected from 0, 1, or 2, and n3 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

7. Said L ２ is an optionally substituted -X １ -(CHROCHR) ｍ２ -C(O)-, where X １ is an optionally substituted aryl group, an optionally substituted heteroaryl group, and preferably, m2 is selected from 0, 1, 2, or 3.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

8. Said L ２ is an optionally substituted (CHR) ｐ 1-C(O)—, where p1 is selected from 0, 1 or 2, and R is H, (CH ２ ) ｎ４ OH, (CH ２ O) ｎ４ (CH ２ CH ２ O) ｎ５ H, and preferably, n4 and n5 are each independently selected from 0, 1, 2, or 3.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

9. Said L ２ is an optionally substituted —(CH ２ ) ｍ１ -X １ - (CH ２ CH ２ O) ｎ３ -(CHR) ｍ２ -C(O)-, where X １ is —C(O)—, and preferably, m1 and m2 are each independently selected from 0, 1, 2, or 3, and n3 is selected from 0, 1, or 2.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

10. Said L ２ is an optionally substituted -X １ - (CH ２ ) ｍ１ -X ２ -(CHR) ｍ２ -C(O)-, where X １ is an optionally substituted aryl group or an optionally substituted heteroaryl group; X ２ is —C(O)—, and preferably, m1 and m2 are each independently selected from 0, 1, or 2.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

11. Said L ２ is -(CHR) ｍ１ -X １ -(CHR) ｍ２ -C(O)-, where X １ is an optionally substituted 3- to 8-membered heteroalicyclic group or an optionally substituted C ３ -C ６ It is an alicyclic group, and preferably, m1 is 0, 1 or 2, and m2 is 0.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

12. Said L ２ is an optionally substituted —(CHR) ｍ１ -X １ - (CH ２ CH ２ O) ｎ３ -(CHR) ｍ２ -C(O)-, where X １ is O, and preferably, m1, n3 and m2 are each independently 0, 1 or 2.

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

13. Said L ２ is an optionally substituted —(CHR) ｍ１ -X １ -(CHR) ｎ３ -X ２ -(CHR) ｍ２ -C(O)-, where X １ is an optionally substituted —C(O)—NR—, and X ２ is O, preferably m1, n3 and m2 are each independently 1, 2 or 3, and R is as defined in claim 2. The ligand conjugate according to claim 2 or its tautomer, meso form, racemic compound, enantiomer, or mixture thereof. Thiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or hydrates thereof.

14. Said L ２ teeth, 【Chemistry 14】 an optionally substituted structure selected from the group consisting of 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

15. Said L ３ is an unsubstituted or CH2-substituted amino acid selected from the group consisting of phenylalanine, isoleucine, leucine, tryptophan, valine, methionine, tyrosine, alanine, threonine, histidine, serine, glutamine, arginine, lysine, asparagine, glutamic acid, proline, citrulline, aspartic acid, and glycine. ２ C(O)R ｃ a peptide residue consisting of an amino acid substituted by 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

16. Said L ３ is an unsubstituted or CH selected from the group consisting of glycine, alanine, lysine, phenylalanine, valine and citrulline. ２ C(O)R ｃ Amino acids substituted by a peptide residue consisting of 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

17. Foreword L ３ are -glycine-phenylalanine-glycine-(-Gly-Phe-Gly-), -glycine-glycine-phenylalanine-glycine-(-Gly-Gly-Phe-Gly-), -valine-citrulline-(-Val-Cit-), -citrulline-valine-(-Cit-Val-), -citrulline-alanine-(-Cit-Ala-), -valine-alanine-(-Val-Ala-), -valine-arginine-(-Val-Arg-), -valine-lysine-(-Val-Lys-), -valine-lysine(Ac)-(-Val-Lys(Ac)-), - Lysine-valine-(-Lys-Val-), -leucine-citrulline-(-Leu-Cit-), -isoleucine-citrulline-(-Ile-Cit-), -tryptophan-citrulline-(-Trp-Cit-), -phenylalanine-lysine-(-Phe-Lys-), -phenylalanine-lysine(Ac)-(-Phe-Lys(Ac)-), -phenylalanine-citrulline-(-Phe-Cit-), -phenylalanine-alanine-(-Phe-Ala-), -phenylalanine-arginine-(-Phe-Arg-), -alanine-lysine -(-Ala-Lys-), -alanine-alanine-(-Ala-Ala-), -alanine-alanine-alanine-(-Ala-Ala-Ala-), -alanine-alanine-asparagine-(-Ala-Ala-Asn-), -alanine-alanine-aspartic acid-(Ala-Ala-Asp-), -lysine-alanine-alanine-asparagine-(-Lys-Ala-Ala-Asn-), -lysine-alanine-alanine-aspartic acid-(-Lys-Ala-Ala-Asp-), -(D)-valine-leucine-lysine-(-D-Val-Leu u-Lys-), -glycine-glycine-arginine-(-Gly-Gly-Arg-), -glycine-glycine-asparagine-(-Gly-Gly-Asn-), -glycine-glycine-phenylalanine-(-Gly-Gly-Phe-), -valine-lysine-glycine-(-Val-Lys-Gly-), -glutamic acid-alanine-alanine-(-Glu-Ala-Ala-), -aspartic acid-alanine-alanine-(-Asp-Ala-Ala-), -valine-lysine-glycine-glycine-(-Val-Lys-Gly-Gly-) and-lysine-alanine-asparagine-(-Lys-Ala-Asn-), unsubstituted or CH ２ C(O)R ｃ a peptide residue substituted by 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

18. Said L ３ teeth, 【Chemistry 15】 【change】 unsubstituted or CH ２ C(O)R ｃ is a structure substituted by 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

19. Said L ４ is a chemical bond, or 【Chemistry 16】 wherein R is an optionally substituted group selected from the group consisting of ａ and R ｂ are each independently hydrogen, optionally substituted C １ -C ４ alkyl groups, optionally substituted C １ -C ４ deuterated alkyl groups 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

20. Said L ４ is a chemical bond, or 【Chemistry 17】 an optionally substituted structure selected from the group consisting of 20. The ligand conjugate of claim 19, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

21. Said L ５ is a chemical bond or an optionally substituted [Chemistry 18] , optionally substituted 【Chemistry 19】 , optionally substituted 【Chemistry 20】 , optionally substituted 【Chemistry 21】 , optionally substituted 【Chemistry 22】 , optionally substituted 【Chemistry 23】 , optionally substituted 【Chemistry 24】 , optionally substituted 【Chemistry 25】 , optionally substituted 【Chemistry 26】 , optionally substituted 【Chemistry 27】 , optionally substituted 【Chemistry 28】 , optionally substituted 【Chemistry 29】 and optionally substituted 【Transformation 30】 The structure is selected from the group consisting of 3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

22. The compound of formula II is characterized in that it is selected from the group consisting of:

3. The ligand complex of claim 2, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof. 【Chemistry 31】

23. The R ４ represents a hydrogen atom, a deuterium atom, a halogen atom, a hydroxyl group, a cyano group, NH ２ , NO ２ , substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ Alkoxy group, substituted or unsubstituted C １ -C ８ alkylthio group, substituted or unsubstituted C １ -C ８ Deuterated alkyl groups, -(CH ２ ) ｍ (C ３ -C ８ cycloalkyl), -(CH ２ ) ｍ (3- to 12-membered heterocyclic group), —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ S(O)(CH ２ ) ｐ R ７ , -(CH ２ ) ｍ S (O) ２ (CH ２ ) ｐ R ７ , -(CH ２ ) ｍ NH (CH ２ ) ｐ R ７ wherein m and p are each independently 0, 1, or 2; ７ is defined as above, R ５ represents a hydrogen atom, a deuterium atom, a halogen, NH ２ , OH, substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ alkoxy groups; Or, R ４ and R ５ together with the carbon atoms to which they are attached, a saturated or unsaturated 5- or 6-membered carbocyclic ring unsubstituted or substituted by one or more Re, an unsubstituted or substituted by one or more R ｅ and forming a structure selected from the group consisting of saturated or unsaturated 5- to 6-membered heterocycles substituted by ｅ The definition of is as described above.

23. The ligand conjugate of claim 22, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

24. R ２ and R ３ are each independently a hydrogen atom, a deuterium atom, a halogen, or NH ２ , substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ Deuterated alkyl groups, -(CH ２ ) ｍ (C ３ -C ６ cycloalkyl), -(CH ２ ) ｍ (3- to 6-membered heterocyclic group), —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ O.C.(O.)R. ７ where m is 0, 1, 2, 3, or 4; ７ is defined as above, Or, R ２ and R ３ together with the carbon atoms to which they are attached, form a structure selected from the group consisting of a saturated or unsaturated 5- to 6-membered ring unsubstituted or substituted by one or more Re, a saturated or unsaturated 5- to 6-membered heterocycle unsubstituted or substituted by one or more Re, wherein the definition of Re is as described above. The ligand conjugate of claim 22 or its tautomer, meso form, racemate, or enamel form. enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or hydrates thereof.

25. The R ４ and R ５ are each independently a hydrogen atom, a deuterium atom, a halogen, a hydroxyl group, or NH ２ , substituted or unsubstituted C １ -C ４ alkyl groups, Or, R ４ and R ５ together with the carbon atoms to which they are attached form an oxa 5-6 membered heterocycle which is unsubstituted or substituted by one or more Re, wherein the definition of Re is as defined above.

23. The ligand conjugate of claim 22, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

26. R ２ is a deuterium atom, a halogen, NH ２ , substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ Deuterated alkyl groups, -(CH ２ ) ｍ (C ３ -C ６ cycloalkyl), -(CH ２ ) ｍ (3- to 6-membered heterocyclic group), —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ O.C.(O.)R. ７ wherein m is 0, 1, 2, 3, or 4; R ３ are each a hydrogen atom, a deuterium atom, a halogen atom, a substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ Deuterated alkyl groups, -(CH ２ ) ｍ (C ３ -C ６ cycloalkyl), -(CH ２ ) ｍ (3- to 6-membered heterocyclic group), —(CH ２ ) ｍ N (R ７ ) ２ , -(CH ２ ) ｍ O.C.(O.)R. ７ wherein m is 0, 1, 2, 3, or 4; Or, R ２ and R ３ together with the carbon atoms to which they are attached form a structure selected from the group consisting of: a saturated or unsaturated 5- to 6-membered ring that is unsubstituted or substituted by one or more Re, a saturated or unsaturated 5- to 6-membered heterocycle that is unsubstituted or substituted by one or more Re; R ４ represents a hydrogen atom, a deuterium atom, a halogen atom, a hydroxyl group, a cyano group, NH ２ , NO ２ , substituted or unsubstituted C １ -C ８ alkyl group, substituted or unsubstituted C １ -C ８ alkoxy groups; R ５ represents a hydrogen atom, a deuterium atom, a halogen atom, a substituted or unsubstituted C １ -C ８ alkyl groups, Or, R ４ and R ５ together with the carbon atoms to which they are attached, represent —OCH ２ O- or -O(CH ２ ) ２ forming a group selected from the group consisting of O—; R ７ represents a hydrogen atom, a deuterium atom, a halogen atom, a substituted or unsubstituted C １ -C ８ selected from the group consisting of alkyl groups, hydroxy groups, amino groups, cyano groups, nitro groups, and mercapto groups; Here, R ｅ The definition of is as described above.

23. The ligand conjugate of claim 22, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

27. The compound of formula II is characterized in that it is selected from the group consisting of:

23. The ligand conjugate of claim 22, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof. 【Chemistry 32】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

28. The Ab is an antibody or an antigen-binding fragment thereof.

2. The ligand conjugate of claim 1, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

29. The antibody is selected from the group consisting of a mouse antibody, a chimeric antibody, a humanized antibody, and a fully human antibody.

29. The ligand conjugate of claim 28, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

30. The antibody is a monoclonal antibody, a bispecific antibody, or a polypeptide.

29. The ligand conjugate of claim 28, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

31. The antigen-binding fragments include Fab, Fab', Fv fragments, F(ab') ２ , F(ab) ２ , scFv, di-scFv, VHH and dAb.

29. The ligand conjugate of claim 28, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

32. The ligand complex is characterized in that it is selected from the group consisting of:

2. The ligand conjugate of claim 1, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof. 【Transformation 33】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

33. The ligand conjugate comprises a structure shown in formula (Ia): 【Transformation 34】 Here, R ４ , R ５ and L are as defined in claim 1, and a is greater than 0. a is a decimal number or an integer.

2. The ligand conjugate of claim 1, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

34. The ligand complex is 【Chemistry 35】 【change】 【change】 【change】 a structure selected from the group consisting of: Here, a is a number greater than 0, and a is a decimal number or an integer.

34. The ligand conjugate of claim 33, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

35. The antibody is characterized in that it is an IGF-1R specific antibody.

35. The ligand complex according to any one of claims 1 to 34, or a tautomer, meso form, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt or hydrate thereof.

36. The IGF-1R is characterized in that it contains IGF-1R derived from a primate.

36. The ligand complex of claim 35.

37. The antibody comprises HCDR3, wherein the HCDR3 comprises the amino acid sequence shown in SEQ ID NO:

3.

37. The ligand complex of claim 36.

38. The antibody comprises HCDR2, wherein the HCDR2 comprises the amino acid sequence shown in SEQ ID NO:

2.

37. The ligand complex of claim 36.

39. The antibody comprises HCDR1, wherein the HCDR1 comprises the amino acid sequence shown in SEQ ID NO:

1.

37. The ligand complex of claim 36.

40. The antibody comprises a heavy chain variable region VH, the VH comprising the HCDR1, HCDR2 and HCDR3, the HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, the HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, and the HCDR1 comprising the amino acid sequence shown in SEQ ID NO:

1.

37. The ligand complex of claim 36.

41. The antibody comprises a heavy chain variable region VH, and the VH comprises the amino acid sequence shown in SEQ ID NO:

4.

37. The ligand complex of claim 36.

42. The antibody is characterized in that it has a full-length sequence as shown in SEQ ID No.

5.

37. The ligand complex of claim 36.

43. 1. A pharmaceutical composition comprising:

43. A pharmaceutical composition characterized in that it comprises a ligand conjugate according to any one of claims 1 to 42, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt, prodrug or solvate thereof, and optionally a pharmaceutically acceptable carrier.

44. Use of a ligand conjugate according to any one of claims 1 to 42, or a tautomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, prodrug, or solvate thereof, and / or a pharmaceutical composition according to claim 43, in the preparation of a medicament for treating and / or preventing a disease or condition associated with target point expression and / or abnormal expression of said ligand.

45. The disease or condition associated with the expression and / or abnormal expression of the target point of the ligand is a tumor, a cancer, an autoimmune disease or an infectious disease, and preferably, the tumor / cancer is a tumor / cancer with high, medium or low expression of the target point of the ligand.

45. The use according to claim 44.

46. A ligand conjugate precursor, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof, The ligand conjugate precursor comprises the structure shown in formula IA: Formula (IA): L Ａ -P Here, L Ａ Is, L １Ａ -L ２ -L ３ -L ４ -L ５ where L １Ａ teeth, 【Transformation 36】 and R ｄ , L ２ , L ３ , L ４ , L ５ and P are defined as in any one of claims 1 to 34, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof.

47. The ligand complex precursor is characterized in that it is selected from the group consisting of:

47. The ligand conjugate precursor of claim 46, or a tautomer, meso form, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt or hydrate thereof. 【Chemistry 37】 【change】 【change】 【change】 【change】

48. A connector as shown in formula (L), It links a Drug unit to a Ligand to form a Ligand-Drug conjugate; Formula (L): L １ -L ２ -L ３ -L ４ -L ５ Here, the L １ , L ２ , L ３ , L ４ , L ５ The connector, characterized in that the definition of is as set forth in any one of claims 1 to 34.

49. The connector is characterized in that it is selected from the group consisting of:

49. The connector of claim 48. 【Transformation 38】 【change】 【change】 【change】 【change】

50. The connector is L １ linked to the ligand via a segment, L ５ and P1 is linked to P1 via a segment to form a ligand-drug conjugate, and P1 is selected from the group consisting of glycopeptide antibiotics such as bleomycin or bleomycin; topoisomerase I inhibitors (e.g., camptothecin, hydroxycamptothecin, 9-aminocamptothecin, SN-38, irinotecan, isotecan, topotecan, belotecan, rubitecan, DXd, etc.), topoisomerase II inhibitors (e.g., actinomycin D, doxorubicin, duocarmycin, daunorubicin, mitomycin, etc.), DNA topoisomerase inhibitors, such as, for example, toxantrone, podophyllotoxin or etoposide; drugs interfering with DNA synthesis, such as, for example, methotrexate, 5-fluorouracil, cytarabine, gemcitabine, mercaptopurine, pentostatin, fludarabine, cladribine or nelarabine; drugs acting on structural proteins, such as, for example, tubulin inhibitors, vinca alkaloids, vincristine, vinblastine, paclitaxel, docetaxel or cabazitaxel; serine / threonine kinase inhibitors tumor signaling pathway inhibitors such as steroid agents, tyrosine kinase inhibitors, aspartate kinase inhibitors, or histidine kinase inhibitors; proteasome inhibitors; histone deacetylase inhibitors; tumor angiogenesis inhibitors; cell cycle protein inhibitors; maytansine derivatives (e.g., DM1, DM4, etc.); calicheamicin derivatives; auristatin derivatives (e.g., monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), auristatin E, auristatin F, etc.); pyrrolobenzodiazepine dimers (pyrrolobenzodiazepine dimers (PBD) derivatives; Amanita mushroom derivatives (such as α-amanitin); anthracyclines; ducalcines; eribulin; melphalan; mitomycin C; chlorambucil; TLR agonists; STING agonists; glucocorticoids, and groups formed by dehydrogenation of other active substances that inhibit tumor cell growth and promote tumor cell apoptosis or necrosis. The connector according to claims 48 to 49.

51. A connector precursor as shown in formula (L-1), which is used to link a Drug unit to a Ligand to form a Ligand-Drug conjugate; Equation (L-1): L Ａ -R ｇ Here, R ｇ is H, OH, O(C １ -C ６ alkyl), where L Ａ 47. The connector precursor, characterized in that the definition of is as set forth in claim 46.

52. The connector precursor is characterized in that it is selected from the group consisting of:

52. The connector precursor of claim 51. 【Chemistry 39】 【change】 【change】 【change】

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