Eribulin derivatives, antibody drug conjugates thereof, and medical uses thereof

Abstract

This application relates to the field of pharmaceutical technology, specifically to eribulin derivatives and their antibody-drug conjugates and pharmaceutical uses. This application involves modifying eribulin to synthesize a series of derivatives with good tumor-suppressive effects, exhibiting lower toxicity and a larger therapeutic window compared to eribulin; then, using suitable linkers, eribulin derivatives are combined with antibodies to form ADCs, utilizing the antibody's targeting specificity to improve the therapeutic efficiency of the eribulin derivatives.

Description

Eribulin derivatives, antibody drug conjugates thereof and medical uses thereof TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a kind of eribulin derivatives, antibody drug conjugates thereof and medical uses thereof. BACKGROUND

[0002] Antibody drug conjugates (ADC) is a kind of therapeutic agent composed of antibody (Antibody) specifically targeting related antigen and small molecule toxin (Payload) coupled by linker (Linker). It has the strong killing effect of small molecule chemotherapeutic agent and the tumor targeting of antibody drug. ADC molecule enters the body, can be combined with the antigen on the surface of target cell through the guiding effect of antibody, and releases toxin molecule through chemical or enzymatic action to achieve the purpose of eliminating target cell. Due to its targeting and drug release in tumor cells, ADCs can accurately bind to tumors and reduce the impact on normal cells, can increase the local concentration of drugs, reduce the toxic side effects on normal cells, and expand the treatment window. ADCs have a wide application prospect in tumor treatment, and have achieved some encouraging results in clinical research.

[0003] The early failure of ADC is mainly the problem of toxicity. Some side effects of ADC are similar to those caused by conventional cytotoxins used in chemotherapy, which are sometimes very serious. One is caused by uncontrolled release of cytotoxin due to instability of linker during blood circulation; Two, the immunoglobulin G1 (IgG1) isotope of some ADCs can interact with (Fc)-gamma receptor (FcgR), which can trigger FcgR-independent internalization in FcgR-positive cells. The off-target toxicity of this antibody will have toxic effects on these healthy cells, which are not the target of ADC. While other side effects are related to the specificity (on-target toxicity) of ADC. According to statistical analysis, the limited dose of ADC is almost consistent with the limited dose of chemotherapeutic drugs as toxins (Cancer Cell 40, 14, 2022, 1257). These factors make the development of cytotoxin of ADC a complex work.

[0004] Eribulin belongs to a class of drugs that inhibit microtubule polymerization, with a unique and novel effect. Eribulin is a synthetic analogue of Halichondrin B. In 1986, Japanese scientists Hirata and Uemura isolated a polyether macrolide compound containing only C, H, and O atoms from the marine sponge Halichondria okadai. They named this extremely complex natural product Halichondrin B. Eribulin mediates antitumor activity by inhibiting microtubule elongation and mitotic spindle formation, leading to apoptosis. At the same time, studies have shown that Eribulin has a subnanomolar effect on both mitotic and non-mitotic tumor and tumor microenvironment. In November 2010, Eribulin was first approved in the United States for the treatment of patients with metastatic breast cancer. Currently, Eribulin has been approved for the treatment of breast cancer in more than 65 countries and regions, including Europe, the Americas and Asia. In addition, Eribulin was first approved in the United States in January 2016 for the treatment of soft tissue sarcoma, and subsequently approved in 55 countries and regions. At the same time, Eribulin has been identified as an orphan drug for the treatment of soft tissue sarcoma in the United States and Japan. The five most common side effects of Eribulin are: decreased white blood cell count, decreased neutrophil count, increased aspartate aminotransferase, increased alanine aminotransferase, and anemia.

[0005] The present application synthesizes a series of derivatives of Eribulin with good tumor inhibition effect by modifying Eribulin, which has lower toxicity and a larger therapeutic window than Eribulin. Then, the Eribulin derivative is connected to an antibody through a suitable linker to make an ADC, which takes advantage of the targeting specificity of the antibody to improve the therapeutic efficiency of the Eribulin derivative. SUMMARY

[0006] To achieve the purpose of the present application, in a first aspect, the present application provides an Eribulin derivative, which is a compound of formula (I) or formula (II), or an isotopically labeled compound thereof, or an optical isomer, geometric isomer, tautomer or isomer mixture thereof, or a pharmaceutically acceptable salt thereof,

[0007] wherein:

[0008] A1 in formula (I) is selected from

[0009] A2 in formula (II) is selected from * and ** are each independently a single bond, a double bond or an oxygen bond =O, and at least one is not a double bond;

[0010] R1is selected from H, alkyl, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, or cyano-substituted alkyl;

[0011] X1is selected from -CO-, -CH2-, -S(O)-, or -S(O)2-;

[0012] X2is selected from a direct bond, -O-, or -NR1-;

[0013] X3is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, or

[0014] Y1is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, or

[0015] R2, R3are each independently selected from H, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl, substituted or unsubstituted carbocyclyl-substituted alkyl, substituted or unsubstituted heterocyclyl-substituted alkyl, substituted or unsubstituted carbocyclyl, or substituted or unsubstituted heterocyclyl; said substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl means optionally substituted with one or more substituents selected from oxo, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl;

[0016] R4is selected from substituted or unsubstituted carbocyclyl or substituted or unsubstituted heterocyclyl; said substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl means optionally substituted with one or more substituents selected from oxo, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl;

[0017] R is selected from substituted or unsubstituted carbocyclyl or substituted or unsubstituted heterocyclyl; said substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl means optionally substituted with one or more substituents selected from oxo, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl;

[0018] ring is a heterocyclyl group, which contains at least one -NH- in the ring and / or which is substituted by amino, hydroxy, amino-substituted alkyl or hydroxy-substituted alkyl, which is optionally substituted by one or more substituents selected from oxo, alkyl, alkyloxy, halogen, cyano, alkyloxy-substituted alkyl, halogen-substituted alkyl, cyano-substituted alkyl;

[0019] m1 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0020] m2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0021] In particular, the alkyl group is a C 1-12 linear or branched alkyl group (e.g. having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12 carbon atoms). The carbocyclyl group can be a saturated, partially unsaturated or fully unsaturated carbocyclyl group, e.g. a cycloalkyl group, an aryl group, a cycloalkenyl group, a cycloalkynyl group, a cycloalkenynyl group, having 3 to 20 carbon atoms (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20 carbon atoms). The heterocyclyl group can be a saturated, partially unsaturated or fully unsaturated heterocyclyl group, e.g. a heterocycloalkyl group, a heteroaryl group, having 3 to 20 atoms (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20 atoms), which can contain 1, 2, 3, 4 heteroatoms selected from N, O, S.

[0022] In one set of embodiments, R1 is selected from H, alkyl, hydroxy-substituted alkyl; preferably from H, C 1-6 alkyl, hydroxy-substituted C 1-6 alkyl; more preferably from H, C 1-4 alkyl, hydroxy-substituted C 1-4 alkyl; in particular from H, methyl, ethyl, n-propyl, i-propyl, hydroxymethyl, hydroxyethyl, hydroxy-n-propyl, hydroxy-i-propyl.

[0023] In one set of embodiments, X1 is selected from -CO-, -CH2-.

[0024] In one set of embodiments, m1 is selected from 1, 2, 3, 4, 5, 6; m2 is selected from 0, 1, 2, 3, 4.

[0025] In one set of embodiments, R2, R3 are each independently selected from H, C 1-6 alkyl, hydroxy, C 1-6 alkyloxy, hydroxy-substituted C 1-6 alkyl, C1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 3-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C

[0026] In one group of embodiments, R4 is selected from substituted or unsubstituted C 3-12 monocyclic, annelated, fused, spiro or bridged cycloalkyl, substituted or unsubstituted 3-12 membered monocyclic, annelated, fused, spiro or bridged heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, substituted or unsubstituted C 6-10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms N or O; the substitution means being substituted with one or more substituents selected from oxo, C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 1-6one or more substituents selected from oxo, C 4-12 monocyclic, annelated, fused, spiro or bridged ring cycloalkyl, 4-12 membered monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, substituted or unsubstituted phenyl, 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms N or O, said substitution means being substituted with one or more substituents selected from oxo, C 1-6 one or more substituents selected from oxo, C

[0027] In one set of embodiments, X3 is selected from -(CR2R3) monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said heterocycloalkyl containing at least one -NH- in the ring, said substitution means being substituted with one or more substituents selected from oxo, C 1-6 alkyl, C 1-6 alkyloxy, C 1-6 alkyloxy substituted C 1-6 one or more substituents selected from oxo, C

[0028] In one set of embodiments, X3 is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH or said monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said substitution means being substituted with one or more substituents selected from oxo, C 1-6 one or more substituents selected from oxo, C 1-6 alkyl, hydroxy, C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted C 3-6 cycloalkyl means optionally substituted with one or more substituents selected from oxo, C 1-6 alkyl; R4 is selected from C 4-12 monocyclic, annelated, fused, spiro or bridged ring cycloalkyl, 4-12 membered monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, phenyl.

[0029] More preferably, X3is selected from the group consisting of:

[0030] More preferably, X3is selected from the group consisting of:

[0031] In one set of embodiments, Y1is selected from the group consisting of: m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, or the ring is a substituted or unsubstituted 4-12 membered monocyclic, fused, polycyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said substitution means substituted with one or more substituents selected from oxo, C 1-6 alkyl; ml is selected from 1, 2, 3, 4, 5, 6; m2 is selected from 0, 1, 2, 3, 4, 5 or 6; R2, R3are each independently selected from H, C 1-6 alkyl, hydroxyl, hydroxyl substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, said substituted or unsubstituted C 3-6 cycloalkyl means optionally substituted with one or more substituents selected from oxo, C 1-6 alkyl; R4is selected from C 4-12 monocyclic, fused, polycyclic, spirocyclic or bridged cyclic cycloalkyl, 4-12 membered monocyclic, fused, polycyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, phenyl.

[0032] Preferably, Y1is selected from the group consisting of: m1 -NH2, -(CR2R3) m1 -OH; ml is selected from 1, 2, 3, 4, 5, 6; R2, R3are each independently selected from H, C 1-6 alkyl, hydroxyl, hydroxyl substituted C 1-6 alkyl.

[0033] More preferably, Y1is selected from the group consisting of:

[0034] In one set of embodiments, ring is a 3-12 membered monocyclic, fused, spiro or bridged heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said heterocycloalkyl containing at least one -NH- in the ring and / or said heterocycloalkyl being substituted by amino, hydroxy, amino-substituted C 1-6 alkyl or hydroxy-substituted C 1-6 alkyl, said heterocycloalkyl being optionally substituted by one or more substituents selected from oxo, C 1-6 alkyl, C 1-6 alkyl, C 1-6 alkyl, said heterocycloalkyl being optionally substituted by one or more substituents selected from oxo, C 1-6 alkyl, said heterocycloalkyl being optionally substituted by one or more substituents selected from oxo, C

[0035] Preferably, ring is a 3-12 membered monocyclic, fused, spiro or bridged heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said heterocycloalkyl containing at least one -NH- in the ring and / or said heterocycloalkyl being substituted by amino, hydroxy, amino-substituted C 1-6 alkyl or hydroxy-substituted C 1-6 alkyl.

[0036] More preferably, ring is selected from the following groups:

[0037] In one set of embodiments, A1, A2are selected from the following groups:

[0038] A2in formula (II) is also selected from

[0039] Preferably, A1, A2are selected from the following groups:

[0040] In particular, the compound of formula (I) is one of the following:

[0041] In particular, the compound of formula (II) is one of the following:

[0042] In a second aspect, the present application provides an eribulin derivative which is a compound of formula (I’), formula (II’), or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or a mixture of isomers thereof, or a pharmaceutically acceptable salt thereof,

[0043] wherein:

[0044] Q is a linker group that can be attached to an antibody or antigen binding fragment;

[0045] L is a linker group connecting Q to A1' or Q to A2';

[0046] A1' is a group of A1 connected to L after removal of H of -NH2, -OH, -NH- on a chain or -NH- on a ring;

[0047] A2' is a group of A2 connected to L after removal of H of -NH2, -OH, -NH- on a chain or -NH- on a ring;

[0048] A1, A2, *, ** are defined as in the preceding formula (I), formula (II).

[0049] In one group of embodiments, when X3, Y1 are independently selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, A1' is a group of A1 connected to L after removal of H of terminal -NH2 or terminal -OH of X3, Y1, and A2' is a group of A2 connected to L after removal of H of terminal -NH2 or terminal -OH of X3, Y1;

[0050] when X3, Y1 are independently selected from , A1' is a group of A1 connected to L after removal of H of -NH- on a ring, and A2' is a group of A2 connected to L after removal of H of -NH- on a ring;

[0051] when A1, A2 are independently selected from , A1' is a group of A1 connected to L after removal of H of -NH- on a ring or -NH2, -OH of a ring substituent, and A2' is a group of A2 connected to L after removal of H of -NH- on a ring or -NH2, -OH of a ring substituent;

[0052] R1, X1, X2, R2, R3, R4, ring, ring, m1, m2 are defined as in the preceding formula (I), formula (II).

[0053] In one set of embodiments, R1is H, A1' is a group that A1 would be connected to L if R1of -NR1- was removed, and A2' is a group that A2 would be connected to L if R1of -NR1- was removed.

[0054] In one set of embodiments, L is -L4-L3-L2-L1-, wherein L1is connected to A1' or A2', and L4is connected to Q;

[0055] L1is selected from a direct bond, wherein is a connection site to A1' or A2', is a connection site to L2;

[0056] L2is a direct bond, one amino acid residue, or a short peptide chain (e.g., 2-10 amino acid residues, which can be a short peptide chain of 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acid residues) formed by two or more amino acid residues, which are selected from natural amino acid residues or unnatural amino acid residues, the N-terminus of which is connected to L3, the C-terminus of which is connected to L1, and L2optionally comprises one or more structures selected from the group consisting of formula (III-1) to formula (III-9):

[0057] s is selected from 1, 2, 3, 4, 5, 6, 7, 8, t is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and R5is selected from H or C 1-8 alkyl; preferably, s is selected from 2, 3, 4, 5, 6, t is selected from 4, 5, 6, 7, 8, and R5is selected from H or C 1-6 alkyl;

[0058] L3is selected from a direct bond, or L3is selected from or r1 is selected from 0, 1, 2, 3, 4, 5, 6, r2 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r3 is selected from 1, 2, 3, 4, 5, 6, r4 is selected from 1, 2, 3, 4, 5, 6, r5 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r6 is selected from 0, 1, 2, 3, 4, 5, 6, r7 is selected from 0, 1, 2, 3, 4, 5, 6, r8 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r9 is selected from 1, 2, 3, 4, 5, 6, r10 is selected from 1, 2, 3, 4, 5, 6, W is selected from O or NR6, R6 is selected from H or C 1-8 alkyl, wherein represents the attachment site to L2, 2 occurrences in the same L3 group represents attachment to L2 at both sites; preferably, r1 is selected from 0, 1, 2, 3, 4, r2 is selected from 4, 5, 6, 7, 8, r3 is selected from 1, 2, 3, 4, r4 is selected from 1, 2, 3, 4, r5 is selected from 4, 5, 6, 7, 8, r6 is selected from 0, 1, 2, 3, 4, r7 is selected from 0, 1, 2, 3, 4, r8 is selected from 1, 2, 3, 4, 5, 6, r9 is selected from 1, 2, 3, 4, r10 is selected from 1, 2, 3, 4, W is selected from O or NR6, R6 is selected from H or C 1-6 alkyl;

[0059] L4 is selected from a direct bond, -CO-, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2) p4 -O-(CH2) p5 -C(O)-, -O-(CH2) p1 -CO-, -O-(CH2CH2O) p2 -(CH2) p3 -CO-, -O-(CH2) p4 -O-(CH2) p5 -C(O)-, p1, p2, p3, p4, p5 are each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, R7 is selected from H or C 1-8alkyl, wherein -CO- is attached to L3; preferably, p1 is selected from 1, 2, 3, 4, 5, 6, p2 is selected from 1, 2, 3, 4, p3 is selected from 1, 2, 3, 4, p4 is selected from 1, 2, 3, 4, p5 is selected from 1, 2, 3, 4, R7 is selected from H or C 1-6 alkyl.

[0060] In one set of embodiments, when A1' is a group A1 attached to L by removal of the H of -NH2, -NH- in the chain or -NH- in the ring, or A2' is a group A2 attached to L by removal of the H of -NH2, -NH- in the chain or -NH- in the ring, L1 is selected from a direct bond, when A1' is a group A1 attached to L by removal of the H of -OH, or A2' is a group A2 attached to L by removal of the H of -OH, L1 is selected from

[0061] In one set of embodiments, L1 is selected from a direct bond,

[0062] In one set of embodiments, L2 is one amino acid residue, or a short peptide chain formed by 2-8 amino acid residues, preferably the amino acid residue is an amino acid residue formed by an amino acid selected from phenylalanine, alanine, glycine, valine, citrulline, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, arginine; more preferably the amino acid residue is an amino acid residue formed by an amino acid selected from phenylalanine, alanine, glycine, valine, citrulline.

[0063] In one set of embodiments, L2 optionally comprises one or more structures selected from formula (III-1), formula (III-9), preferably optionally comprises one or more structures selected from one or more of the following structures: t = 8.

[0064] In one set of embodiments, L2 is selected from NH Gly CO -, or from the following short peptide chains: NH Gly-Gly-Phe-Gly CO -, NH Val-Cit CO -, NH Val-Ala CO -, NH Gly-Gly-Val-AlaCO -、 NH Gly-Gly-Val-Cit CO -、 NH Lys-Gly-Val-Ala CO -、 NH Lys-Gly-Val-Cit CO -、 NH Gly-Val-Cit CO - and L2optionally comprises one or more structures selected from the group consisting of formula (III-1) to formula (III-9), preferably L2optionally comprises one or more structures selected from the group consisting of formula (III-1), formula (III-9).

[0065] Preferably, L2is selected from the group consisting of:

[0066] Preferably, L2is selected from the group consisting of:

[0067] In one set of embodiments, L3is selected from the group consisting of a direct bond, or or L3is selected from the group consisting of

[0068] or r1, r6, r7are selected from 0, r2is selected from 4, 5, 6, 7, 8, r3is selected from 1, 2, 3, 4, r4is selected from 1, 2, 3, 4, r5is selected from 4, 5, 6, 7, 8, r8is selected from 1, 2, 3, 4, r9is selected from 1, 2, 3, 4, r10is selected from 1, 2, 3, 4, W is selected from NH; preferably L3is selected from a direct bond, r1, r7are selected from 0, r2is selected from 4, 5, 6, 7, 8, r3is selected from 2, r8is selected from 2, 3, 4, r9is selected from 2, r10is selected from 2.

[0069] In one set of embodiments, L4is selected from the group consisting of a direct bond, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2)p4 -O-(CH2) p5 -C(O)-, p1 is selected from 1, 2, 3, 4, 5, 6, p2, p3, p4, p5 are each independently selected from 1, 2, 3, 4, R7 is selected from H or C 1-6 alkyl. Preferably, L4 is selected from a direct bond, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2) p4 -O-(CH2) p5 -C(O)-, p1 is selected from 3, 4, 5, 6, p2, p3, p4, p5 are each independently selected from 2, 3, R7 is selected from H.

[0070] In one set of embodiments, Q is selected from a group comprising a maleimide linker, a methanesulfonylpyrimidine linker, a methanesulfonylpyridothiazole linker, a methanesulfonylpyrimidinethiazole linker, a cycloalkynyl alkyne linker, or an oxime bond linker.

[0071] In one set of embodiments, Q is selected from

[0072] In one set of embodiments, Q-L4 is selected from p1 is selected from 1, 2, 3, 4, 5, 6, p2 is selected from 1, 2, 3, 4, p3 is selected from 1, 2, 3, 4, p4 is selected from 1, 2, 3, 4, p5 is selected from 1, 2, 3, 4, R7 is selected from H or C 1-6 alkyl. Preferably, p1 is selected from 3, 4, 5, 6, p2 is selected from 2, 3, p3 is selected from 2, 3, p4 is selected from 2, 3, p5 is selected from 2, 3, R7 is selected from H.

[0073] Preferably, Q-L4 is selected from

[0074] In particular, formula (I’) is one of the following:

[0075] In particular, formula (II’) is one of the following:

[0076] In a third aspect, the present application provides an antibody drug conjugate, which is a compound of formula (I”), formula (II”), or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof,

[0077] wherein:

[0078] Ab is an antibody or an antigen binding fragment;

[0079] L5 is a direct bond or a linker connecting Ab and Q’;

[0080] Q’ is a group of Q after connecting with Ab through L5;

[0081] n is an integer or a decimal number between 0 and 20; such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 0 and a decimal number between each of the above integers;

[0082] Q, L, A1’, A2’, *, ** are defined as in formula (I’), formula (II’) above.

[0083] In one group of embodiments, n is an integer or a decimal number between 0 and 8.

[0084] In one group of embodiments, L5 is a direct bond or represents a connection site with Ab, represents Z3 connecting with Q’; Z3 is a click probe or a thiol group or a precursor thereof capable of connecting with Q’ after a reaction, such as a metal-free click reaction, preferably Z3 is an azido group;

[0085] Z1 and Z2 are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NHC(O)-, -NH-, -O-, -OC(O)NH- or -NHC(O)O-;

[0086] q1 is 0, 1, 2, 3, 4, 5, 6, 7, 8;

[0087] q2 is 1, 2, 3, 4, 5, 6, 7, 8;

[0088] q3 is 1, 2, 3, 4, 5, 6, 7, 8;

[0089] q4 is 1, 2, 3, 4, 5, 6, 7, 8.

[0090] Preferably, L5 is a direct bond or More preferably, L5 is a direct bond or

[0091] Preferably, Z1and Z2are each independently selected from -C(O)NH-.

[0092] In one set of embodiments, Q' is selected from:

[0093] represents a connection site for L5, represents a connection site for L4.

[0094] In one set of embodiments, L5-Q' is selected from: represents a connection site for Ab, represents a connection site for L4.

[0095] Preferably, L5-Q' is selected from More preferably, Z1and Z2are each independently selected from -C(O)NH-.

[0096] In one set of embodiments, the Ab is selected from a murine antibody, a chimeric antibody, a humanized antibody, a fully human antibody, or an antigen-binding fragment thereof; preferably, the Ab is selected from an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-ROR1 antibody, an anti-CLDN6 antibody, an anti-CLDN9 antibody, an anti-CLDN18.2 antibody, an anti-NaPi-2b antibody, an anti-TNF-a antibody, an anti-ENPP3 antibody, an anti-DLL3 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD28 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD37 antibody, an anti-CD38 antibody, an anti-CD44 antibody, an anti-CD45 antibody, an anti-CD47 antibody, an anti-CD48 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD98 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-EphA2 antibody, an anti-MUC1 antibody, an anti-Lewis Y antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, an anti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-C antibody, an anti-SLC44A4 antibody, an anti-CD79 antibody, an anti-TROP-2 antibody, an anti-CD79B antibody, an anti-Mesothelin antibody, an anti-Nectin-4 antibody, an anti-TPBG antibody, or an antigen-binding fragment thereof.

[0097] Preferably, wherein the Ab is an anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof.

[0098] Preferably, wherein the Ab is selected from a monoclonal antibody or an antigen-binding fragment thereof, preferably selected from Trastuzumab, Cetuximab, Pertuzumab, Nimotuzumab, Enoblituzumab, Emibetuzumab, Inotuzumab, Pinatuzumab, Brentuximab, Gemtuzumab, Bivatuzumab, Lorvotuzumab, or an antigen-binding fragment thereof; more preferably, wherein the Ab is selected from Trastuzumab or an antigen-binding fragment thereof.

[0099] In one set of embodiments, Formula (I”) is one of the following:

[0100] ; preferably, Ab is an anti-HER2 (ErbB2) antibody or antigen-binding fragment thereof, more preferably Trastuzumab or an antigen-binding fragment thereof.

[0101] In one group of embodiments, formula (II") is one of the following:

[0102] ; preferably, Ab is an anti-HER2 (ErbB2) antibody or antigen-binding fragment thereof, more preferably Trastuzumab or an antigen-binding fragment thereof.

[0103] In a fourth aspect, the present application provides an eribulin derivative, which is a compound as shown below, or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or a mixture of isomers thereof, or a pharmaceutically acceptable salt thereof,

[0104] In a fifth aspect, the present application provides an antibody drug conjugate, which is a compound as shown below, or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or a mixture of isomers thereof, or a pharmaceutically acceptable salt thereof,

[0105] ; Ab, n are defined as defined in formula (I"), formula (II") above. Preferably, Ab is an anti-HER2 (ErbB2) antibody or antigen-binding fragment thereof, more preferably Trastuzumab or an antigen-binding fragment thereof.

[0106] In a sixth aspect, the present application provides a pharmaceutical composition comprising an eribulin derivative, an antibody drug conjugate according to the present application above, and one or more pharmaceutically acceptable carriers or excipients.

[0107] In a seventh aspect, the present application provides use of an eribulin derivative according to the present application above in the manufacture of an antibody drug conjugate.

[0108] In an eighth aspect, the present application provides use of an eribulin derivative, an antibody drug conjugate, a pharmaceutical composition according to the present application above in the manufacture of a medicament for treating a tumor or cancer.

[0109] In a ninth aspect, the present application provides a method for treating a tumor or cancer, comprising administering to a subject in need thereof an effective amount of the eribulin derivative, the antibody drug conjugate, or the pharmaceutical composition according to the preceding aspects of the present application.

[0110] In one group of embodiments, the tumor or cancer is selected from a solid tumor or a hematological tumor; preferably, the tumor or cancer is selected from a metastatic, recurrent tumor or cancer; preferably, the tumor or cancer is selected from breast cancer, gastric cancer, colon cancer, pancreatic cancer, renal cancer, ovarian cancer, lung cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, melanoma, head and neck cancer, cervical cancer, prostate cancer; in particular, the tumor or cancer can be HER-2 positive breast cancer, triple negative breast cancer, HER-2 positive triple negative breast cancer, renal clear cell adenocarcinoma, small cell lung cancer, non-small cell lung cancer, large cell lung cancer.

[0111] In one group of embodiments, the tumor or cancer is selected from a cancer or tumor with high expression of HER2 (ErbB2) antibody.

[0112] The present application synthesizes a series of derivatives of eribulin with good tumor inhibition effect by modifying eribulin, which has lower toxicity and a larger therapeutic window than eribulin, and further improves the therapeutic efficiency of the eribulin derivative by using the targeting specificity of the antibody to make an ADC of the eribulin derivative and the antibody. BRIEF DESCRIPTION OF DRAWINGS

[0113] Figure 1 is a tumor volume change curve of JIMT-1 breast cancer tumor-bearing mouse model-1.

[0114] Figure 2 is a body weight change curve of rats after administration.

[0115] Figure 3 is a tumor volume change curve of JIMT-1 breast cancer tumor-bearing mouse model-2. DETAILED DESCRIPTION

[0116] Unless otherwise defined, all terms (including technical and scientific terms) used in disclosing the present application have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The following definitions are set forth to facilitate the understanding of the present application.

[0117] The features described or illustrated as part of one embodiment in the present application can be used in another embodiment to produce a further embodiment.

[0118] The selection scope of the terms "and / or", "or / and", "and / or" used in the present application includes any one of the two or more related listed items, and also includes any and all combinations of the related listed items, including any two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", "and / or", it should be understood that in the present application, the technical solution undoubtedly includes the technical solution connected by "logical and", and also undoubtedly includes the technical solution connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For another example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C and D (i.e. the technical solution connected by "logical or"), and also includes any and all combinations of A, B, C and D, i.e. includes the combination of any two or any three of A, B, C and D, and also includes the four-item combination of A, B, C and D (i.e. the technical solution connected by "logical and").

[0119] The terms "containing", "including" and "comprising" used in the present application are synonymous and are inclusive or open-ended, and do not exclude additional, unrecited members, elements or method steps.

[0120] The numerical ranges represented by endpoints in the present application include all numbers and fractions within the range, and the recited endpoints.

[0121] In the present application, the concentration values involved include fluctuations within a certain range. For example, it can fluctuate within a corresponding accuracy range. For example, 2% can be allowed to fluctuate within ±0.1%. For larger values or values that do not need to be controlled too finely, it is also allowed that the meaning includes larger fluctuations. For example, 100mM can be allowed to fluctuate within ±1%, ±2%, ±5%, etc. For molecular weight, it is allowed that the meaning includes a fluctuation of ±10%.

[0122] In the present application, descriptions such as "multiple", "various" and the like are used without special limitation, which means greater than or equal to 2 in quantity.

[0123] In the present application, among the technical features described in an open-ended manner, both the closed technical solution consisting of the listed features and the open technical solution containing the listed features are included.

[0124] In the present application, "preferably", "more preferably", "most preferably", "particularly", "more particularly" or "most particularly" are used to describe the embodiments or examples of the present application, and are not used to limit the scope of the present application. In the present application, "optionally", "optionally" or "optional" means that it can or can not be present, i.e. it means that it is selected from "yes" or "no" two parallel schemes. If there are multiple "options" in a technical solution, each "option" is independent of each other unless otherwise specified, and there is no contradictory relationship or mutual restriction.

[0125] The eribulin derivatives described in the present application include the compounds of formula (I), the compounds of formula (II), the compounds of formula (I'), the compounds of formula (II'), the compound of claim 44, the isotopically labeled compounds of the compounds of formula (I), the compounds of formula (II), the compounds of formula (I'), the compounds of formula (II'), the compound of claim 44, the optical isomers, geometric isomers, tautomers or isomer mixtures of the compounds of formula (I), the compounds of formula (II), the compounds of formula (I'), the compounds of formula (II'), the compound of claim 44 or the isotopically labeled compounds, and the pharmaceutically acceptable salts of the above various compounds.

[0126] The antibody drug conjugates described in the present application include the compounds of formula (I"), the compounds of formula (II"), the compounds of claims 45-48, the isotopically labeled compounds of the compounds of formula (I"), the compounds of formula (II"), the compounds of claims 45-48, the optical isomers, geometric isomers, tautomers or isomer mixtures of the compounds of formula (I"), the compounds of formula (II"), the compounds of claims 45-48 or the isotopically labeled compounds, and the pharmaceutically acceptable salts of the above various compounds.

[0127] The term "isotopically labeled compound" refers to an isotopically labeled compound obtained by replacing any atom in the compound with its isotope atom. All pharmaceutically acceptable isotopically labeled compounds of the compounds described in the present application, wherein one or more atoms are replaced by atoms having the same atomic number but different atomic mass or mass number as usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the present application include isotopes of hydrogen, such as 2 H(D) and 3 H(T), isotopes of carbon, such as 11 C, 13 C and 14 C, isotopes of chlorine, such as 37 Cl, isotopes of fluorine, such as 18 F, isotopes of iodine, such as 123 I and 125 I, isotopes of nitrogen, such as13 N and 15 N, isotopes of oxygen, such as 15 O, 17 O and 18 O, and isotopes of sulfur, such as 35 S. Isotopically-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples and Preparations herein using an appropriate isotopically-labeled reagents in place of the non-labeled reagent previously employed.

[0128] The term "optical isomer" means, when a compound has one or more chiral centers, each chiral center can exist in either the R or S configuration, the various isomers thus constituted are optical isomers. Optical isomers include all diastereomeric, enantiomeric, meso, racemic or mixtures thereof. Optical isomers can be separated, for example, by a chiral chromatographic column or by chiral synthesis.

[0129] The term "geometric isomer" means, when a compound has a double bond, the compound can exist as a cis isomer, trans isomer or E isomer, Z isomer. Geometric isomers include cis isomer, trans isomer, E isomer, Z isomer or mixtures thereof.

[0130] The term "tautomers" means isomers that result from the rapid movement of a certain atom in the molecule between two positions. Those skilled in the art can understand that tautomers can be converted to each other, and can coexist in a certain state in a state of equilibrium.

[0131] The compounds of the present application can exist in pharmaceutically acceptable salt forms, such as acid addition salts and / or base addition salts of the compounds of Formula (I). As used herein, "pharmaceutically acceptable salt" includes an acid addition salt or a base addition salt that can occur as a result of the compounds of the present application. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples of such acids include, but are not limited to: acetic, adipic, aspartic, benzoic, benzenesulfonic, bicarbonic / carbonic, bisulfic, boric, camphorsulfonic, citric, cyclamic, ethanesulfonic, formic, fumaric, gluconic, glucoheptonic, glucuronic, hexafluorophosphoric, 2-(4-hydroxybenzoyl)benzoic, hydrochloric / chloride, hydrobromic / bromide, hydroiodic / iodide, 2-hydroxyethanesulfonic, lactic, malic, maleic, malonic, methanesulfonic, sulfuric, methylsulfate, naphthalenesulfonic, 2-naphthalenesulfonic, nicotinic, nitric, orotic, oxalic, palmitic, phosphoric / dihydrogen phosphate / bi hydrogen phosphate, pyroglutamic, saccharic, stearic, salicylic, tannic, tartaric, toluenesulfonic, and trifluoroacetic. Suitable base addition salts are formed from bases which form non-toxic salts. Examples of such bases include, but are not limited to: aluminum, arginine, calcium, choline, diethylamine, diethanolamine, glycine, lysine, magnesium, meglumine, ethanolamine, potassium, sodium, tromethamine, and zinc salts. Acid and base half-salts can also be formed, such as hemisulfates and hemicalcium salts. For a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection and Use by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds described herein are known to those skilled in the art.

[0132] Certain compounds of the present application can exist in different crystalline or amorphous forms, all of which are intended to be within the scope of the present application.

[0133] In the present application, the term "antibody" refers to an immunoglobulin, which is a four polypeptide chain structure connected by interchain disulfide bonds, consisting of two identical heavy chains and two identical light chains. The amino acid composition and arrangement of the constant region of the heavy chain of immunoglobulin are different, and so are their antigenicities. Accordingly, immunoglobulins can be classified into five types, or isotypes, namely IgM, IgD, IgG, IgA and IgE, with their corresponding heavy chains being μ chain, δ chain, γ chain, α chain and ε chain, respectively. The same type of Ig can be further classified into different subtypes according to the differences in the amino acid composition of the hinge region and the number and position of the disulfide bonds of the heavy chain, such as IgG1, IgG2, IgG3 and IgG4. The light chain can be classified into κ chain or λ chain according to the constant region. Each of the five types of Ig can have κ chain or λ chain. The sequences of about 110 amino acids at the N-terminus of the heavy chain and light chain of the antibody vary greatly, which are the variable region (Fv region); the remaining amino acid sequences at the C-terminus are relatively stable, which are the constant region. The variable region includes three hypervariable regions (HVR) and four relatively conserved framework regions (FR). The three hypervariable regions determine the specificity of the antibody, also known as the complementarity determining region (CDR). Each light chain variable region (LCVR) and heavy chain variable region (HCVR) consists of three CDR regions and four FR regions, arranged in the order of FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4 from the amino terminus to the carboxyl terminus. The three CDR regions of the light chain are LCDR1, LCDR2 and LCDR3; the three CDR regions of the heavy chain are HCDR1, HCDR2 and HCDR3. The CDR amino acid residues of the LCVR region and the HCVR region of the antibody or antigen binding fragment described in the present application are in accordance with the known Kabat numbering rule (LCDR1-3, HCDR2-3), or in accordance with the numbering rule of kabat and chothia (HCDR1).

[0134] The term "antigen-binding fragment" refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding fragment" include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL, and CH domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fv fragment consisting of the VHand VLdomains of a single arm of an antibody; (iv) a Fv fragment consisting of the VHand VLdomains of a single arm of an antibody; (v) a single domain or dAb fragment, which consists of a VHand; and (vi) an isolated complementarity determining region (CDR) or (vii) a combination of two or more isolated CDRs, which can optionally be joined by a synthetic linker. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be produced as a single protein chain (known as single chain Fv (scFv)), in which the VL and VH regions pair to form monovalent molecules. Such single chain antibodies are also included within the term "antigen-binding fragment". Such antibody fragments are obtained using conventional techniques known to those with skill in the art, and are screened for utility in the same fashion as are intact antibodies. Antigen binding moieties can be produced by recombinant DNA techniques, or by enzymatic or chemical cleavage of intact immunoglobulin. The antibody can be an antibody of different isotypes, e.g., IgG (e.g., IgG1, IgG2, IgG3, or IgG4 subtypes), IgA1, IgA2, IgD, IgE, or IgM antibodies.

[0135] The term "linker group" refers to a chemical moiety or bond that links one end to an antibody and the other end to a toxin drug, which can also be linked to other linker groups before linking to the antibody or toxin drug.

[0136] The term "antibody drug conjugate" (ADC) refers to an antibody or antigen binding fragment linked to a biologically active toxic drug, such as a eribulin derivative, through a stable linking unit.

[0137] The term "short peptide" has its ordinary meaning in the art and can refer to a compound covalently linked together by condensation-dehydration of the carboxyl of one amino acid molecule with the amino of another amino acid molecule to form an amide bond (peptide bond). The nomenclature used to define the peptide is commonly used in the art, where the amino group at the N-terminus appears on the left and the carboxyl group at the C-terminus appears on the right. The short peptide chain described in the present application refers to a peptide chain resulting from the condensation-dehydration of two or more amino acids (such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12), with the -NH- on the left formed by the N-terminal amino group and the -CO- on the right formed by the C-terminal carboxyl group. For example, "Gly-Gly-Phe-Gly" represents a tetrapeptide chain resulting from the sequential condensation of Gly, Gly, Phe, Gly, with the -NH- on the left formed by the N-terminal Gly amino group and the -CO- on the right formed by the C-terminal Gly carboxyl group. NH Gly-Gly-Phe-Gly CO -” represents a tetrapeptide chain resulting from the sequential condensation of Gly, Gly, Phe, Gly, with the -NH- on the left formed by the N-terminal Gly amino group and the -CO- on the right formed by the C-terminal Gly carboxyl group.

[0138] The term "amino acid residue" has its ordinary meaning in the art and refers to the group of an amino acid (as a single amino acid or as part of a peptide) after its reaction with a peptide, another amino acid or amino acid residue. Typically, when an amino acid is combined with another amino acid or amino acid residue, water is removed and the remaining amino acid is referred to as an amino acid residue.

[0139] The term "amino acid" has its ordinary meaning in the art and can include natural amino acids and non-natural amino acids. The abbreviations of the amino acid residues in the present application are the standard 3-letter and / or 1-letter codes used in the art to refer to one of the 20 commonly used amino acids.

[0140] Amino acid residues or amino acids have isomeric forms, including L-forms and D-forms.

[0141] Examples of the three-letter codes and one-letter codes of the amino acids used in the present application and their structures are shown in the following table:

[0142] In some of the substituents in the present application represents a connection site, represents a connection site. For example, is a connection site for connection with A1' or A2', is a connection site for connection with L2, which means that the left side is connected with L2 through -NH and the right side is connected with A1' or A2' through -CH2-.

[0143] Unless otherwise indicated, "optionally substituted" means unsubstituted or substituted by one or more substituents independently selected from one or more of deuterium, oxo, halogen, amino, hydroxyl, nitro, cyano, thiol, optionally substituted alkyl, optionally substituted alkyl oxy, optionally substituted alkyl thio, optionally substituted alkyl amino, optionally substituted carbocyclyl, optionally substituted heterocyclyl; the substituents on alkyl, carbocyclyl, heterocyclyl are selected from one or more of oxo, deuterium, halogen, amino, hydroxyl, nitro, cyano, thiol, alkyl, haloalkyl, alkyloxy, hydroxyl substituted alkyl, alkyloxyalkyl, etc. The case where the substituents are selected from "oxo" means that two hydrogen atoms on the same substituted carbon are replaced by an oxygen atom, or a heteroatom is attached to oxygen, such as nitro oxide where nitrogen is attached to oxygen, sulfinyl or sulfonyl where sulfur is attached to oxygen. "Optionally oxo" means unsubstituted or two hydrogen atoms on the same substituted carbon are replaced by an oxygen atom, or a heteroatom is attached to oxygen, such as nitro oxide where nitrogen is attached to oxygen, sulfinyl or sulfonyl where sulfur is attached to oxygen. The term "independently" means that when the number of substituents is more than one, the substituents can be the same or different.

[0144] The term "oxo" (=0) means that two hydrogen atoms on the carbon atom to which it is attached are replaced by an oxygen atom, forming a carbonyl group.

[0145] In the present application means that N forms a ring with the additional group to which it is attached. means that X3or Y1is a ring containing at least one -NH- in the ring.

[0146] The term "alkyl" refers to saturated aliphatic hydrocarbon groups which are straight-chain or branched groups, preferably containing 1 to 20 carbon atoms (number of carbon atoms between 1 and 12, in particular 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12), more preferably 1 to 10 carbon atoms, most preferably 1 to 8 carbon atoms (i.e. Ci-8alkyl, number of carbon atoms between 1 and 8, in particular 1, 2, 3, 4, 5, 6, 7 or 8), 1 to 6 carbon atoms (i.e. Ci-6alkyl, number of carbon atoms between 1 and 6, in particular 1, 2, 3, 4, 5 or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, sec-butyl, n-pentyl, neopentyl, 1,1 -dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1 -ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, 3-hexyl, 2-pentyl and the like.

[0147] The term "alkyloxy" refers to -O-alkyl, the alkyl group being as defined above. Representative examples include, but are not limited to, methyloxy, ethyloxy, propyloxy, isopropyloxy, butyloxy, 1 -methylpropyloxy, 2-methylpropyloxy, t-butyloxy and the like.

[0148] The term "halogen" refers to F, CI, Br, I.

[0149] The terms "hydroxy-substituted alkyl", "alkyloxy-substituted alkyl", "amino-substituted alkyl", "haloalkyl", "cyano-substituted alkyl", "carbocyclyl-substituted alkyl", "heterocyclyl-substituted alkyl" mean that the alkyl group is substituted by one or more hydroxy groups, the alkyl group is substituted by one or more alkyloxy groups, the alkyl group is substituted by one or more amino groups, the alkyl group is substituted by one or more halogen atoms, the alkyl group is substituted by one or more cyano groups, the alkyl group is substituted by one or more carbocyclyl groups, the alkyl group is substituted by one or more heterocyclyl groups.

[0150] The term "carbocyclyl" refers to saturated, partially unsaturated or fully unsaturated carbocyclic groups, such as cycloalkyl, aryl, cycloalkenyl, cycloalkynyl, cycloalkenynyl groups, which can be monocyclic or polycyclic (including bicyclic, tricyclic, etc.), including fused, annulated, bridged, spiro, bridged, etc. The number of carbon atoms can be 3 to 20 (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20).

[0151] The term "cycloalkyl" refers to saturated monocyclic or polycyclic (including bicyclic, tricyclic, etc.) cyclic hydrocarbon substituents, including single ring, fused ring, spiro ring, bridged ring, etc. Cycloalkyl groups can contain 3 to 20 carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20), preferably 3 to 15 carbon atoms, more preferably 3 to 12 carbon atoms. Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,

[0152] The term "aryl" denotes a monocyclic, bicyclic aromatic carbocyclic ring system containing 6-10 carbon atoms, examples of aryl groups include phenyl, naphthyl, e.g., 1-naphthyl, 2-naphthyl, 3-naphthyl, 4-naphthyl.

[0153] The term "heterocyclyl" refers to saturated, partially unsaturated, or fully unsaturated heterocyclic groups, such as heterocycloalkyl, heteroaryl, which can be monocyclic or polycyclic (including bicyclic, tricyclic, etc.), including single ring, fused ring, spiro ring, bridged ring, etc. The number of atoms can be 3-20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20), and can contain 1, 2, 3, 4 heteroatoms selected from N, O, S, e.g., 1, 2, or 3 heteroatoms selected from N or O.

[0154] ​The term "heterocycloalkyl" refers to saturated monocyclic or polycyclic (including bicyclic, tricyclic, etc.) hydrocarbon substituents, including monocyclic, annelated, fused, spirocyclic, bridged, etc., which contain 3 to 20 ring atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20), wherein one or more ring atoms (e.g., 1, 2, 3, 4) are heteroatoms selected from N, O, or S. Preferably, 3 to 12 ring atoms (including 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms) are contained, wherein 1 to 3 are heteroatoms (1, 2, or 3 heteroatoms) selected from N, O, or S. The heterocycloalkyl group can be a 3-11 membered monocyclic, annelated, fused, spirocyclic, bridged heterocycloalkyl group selected from 1 or 2 N atoms, e.g., 3-6 membered monocyclic heterocycloalkyl, 6-12 membered annelated heterocycloalkyl, 6-12 membered fused heterocycloalkyl, 6-12 membered spirocyclic heterocycloalkyl, 6-12 membered bridged heterocycloalkyl, non-limiting examples including pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, aziridinyl, oxiridinyl, thiiridinyl, azetidinyl, oxetidinyl, thietidinyl, oxepanyl, morpholinyl, thiomorpholinyl, dioxanyl, dithianyl, oxazolidinyl, thiazolidinyl, pyrazolidinyl, imidazolidine, tetrahydrofuran-1-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydropyran-1-yl, tetrahydropyran-2-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, piperidin-1-yl, piperidin-4-yl, etc.

[0155] The term "heteroaryl" denotes an aromatic monocyclic or polycyclic ring system containing a 5-10 membered structure, or preferably a 5-8 membered structure or a 6-10 membered structure, more preferably a 5-6 membered structure, wherein 1, 2, 3, 4 or more ring atoms are heteroatoms and the remaining atoms are carbon, the heteroatoms being independently selected from O, N or S, the number of heteroatoms preferably being 1, 2, 3 or 4. The heteroaryl group can be an aromatic monocyclic ring containing 1-2 5-6 membered structures selected from N, S, O, or an aromatic fused ring containing 1, 2, 3 or 4 8-10 membered structures selected from N, S, O. Examples of heteroaryl groups include, but are not limited to, furanyl, thienyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiadiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazo[l,2-a]pyridyl, pyrazolo[l,5-a]pyridyl, pyrazolo[l,5-a]pyrimidyl, imidazo[l,2-b]pyridazinyl, [l,2,4]triazolo[4,3-b]pyridazinyl, [l,2,4]triazolo[l,5-a]pyrimidyl, [l,2,4]triazolo[l,5-a]pyridyl, pyridin-1-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, quinolin-2-yl, quinolin-3-yl, quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, quinolin-7-yl, quinolin-8-yl, and the like.

[0156] The term "fused ring" means that two rings share two adjacent ring atoms. For example, and the like.

[0157] The term "fused ring" means that two rings share two adjacent ring atoms. For example, and the like.

[0158] The term "spiro ring" means that two rings share one ring atom. For example, and the like.

[0159] The term "bridged ring" means that two rings share two non-adjacent ring atoms. For example, and the like.

[0160] In the present application, the numerical range in connection with the number of substituents, the number of carbon atoms, the number of ring atoms, the number of heteroatoms, the number of groups means the enumeration of all integers within the range, and the range is only a kind of simplified representation. For example, "1-4 substituents" means 1, 2, 3 or 4 substituents; "3-8 ring atoms" means 3, 4, 5, 6, 7 or 8 ring atoms. Therefore, the numerical range in connection with the number of substituents, the number of carbon atoms, the number of ring atoms, the number of heteroatoms, the number of groups also covers any one of its sub-ranges, and each sub-range is also considered to be disclosed in the present application.

[0161] The term "pharmaceutical composition" means a mixture of one or more compounds described in the present application and one or more pharmaceutically acceptable carriers or excipients. The pharmaceutical composition can not contain other active ingredients, or further contain one or more other active ingredients. The purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and thus exert the biological activity.

[0162] The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes, but is not limited to, any of the adjuvants, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersing agents, suspending agents, stabilizers, isotonic agents, solvents, emulsifiers, disintegrating agents, binders, fillers, antioxidants, aromatics, co-solvents, solubilizers, osmotic pressure regulators, base portion in ointments, cream, and the like, which are permitted for use in humans or domestic animals by the relevant government regulatory agencies.

[0163] The pharmaceutical composition described in the present application can be prepared into liquid dosage forms, solid dosage forms or semi-solid dosage forms. The liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w type, w / o type and multiple emulsions), suspensions, injections (including water injections, powder injections and infusions), eye drops, nose drops, lotions and liniments, etc.; the solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, chewing tablets, dispersible tablets, effervescent tablets, oral disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pills, suppositories, films, patches, aerosols, sprays, etc.; the semi-solid dosage forms can be ointments, gels, pastes, etc. The pharmaceutical composition of the present application can be prepared into ordinary preparations, but also into sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0164] The term "subject" refers to any human or non-human organism that can potentially benefit from the treatment with the compounds described in the present application. Exemplary subjects include any age human or mammal. Preferably, the subject is a human.

[0165] The term "treatment" includes treatment of a disease or condition in a mammal, particularly in a human, and includes: (a) inhibiting the infection, disease or condition, i.e., arresting or reducing the development of the infection, disease or condition; (b) relieving the infection, disease or condition, i.e., causing the regression of the disease or condition, and / or (c) curing the infection, disease or condition.

[0166] The term "prevention" includes prophylactic treatment in a mammal, particularly in a human, aimed at reducing the likelihood of the occurrence of an infection, disease or condition. Patients selected for prophylactic treatment can be selected on the basis of an increased risk of infection or of having a disease or condition as compared to the general population. "Prevention" can include treatment of a subject who has not yet presented an infection or clinical condition, and prevention of a second occurrence of the same or similar infection or clinical condition.

[0167] "Effective amount" means an amount of a compound of the application effective to treat or prevent cancer, to inhibit cancer cell proliferation, and / or to reduce tumor volume, when administered alone or in combination.

[0168] Embodiments of the application will be described in detail below with reference to Examples. It should be understood that these Examples are intended to illustrate the application and are not intended to limit the scope of the application. Various modifications and changes can be made by persons of ordinary skill in the art to the application without departing from the scope or spirit of the application. The experimental methods in the following Examples, where no specific conditions are mentioned, are preferably according to the indications given in the application, but can also be according to the protocols or general conditions in the art, or according to other experimental methods known in the art, or according to the conditions suggested by the manufacturer.

[0169] In the following specific examples, the measurement parameters of the raw materials components, if not specified, can have slight deviations within the range of the weighing accuracy. The temperature and time parameters allow for acceptable deviations caused by the accuracy of the instruments or the accuracy of the operation.

[0170] In addition, some protecting groups can be used in the application to protect certain functional groups from unwanted reactions. Suitable protecting groups for various functional groups and their protecting or deprotecting conditions are well known to those skilled in the art. For example, T.W. Greene and G.M. Wuts, Protective Groups in Organic Synthesis (3rded., Wiley, New York, 1999 and references therein) describe a large number of protecting groups in detail.

[0171] The isolation and purification of the compounds and intermediates are carried out by appropriate methods and procedures according to the specific requirements, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin layer chromatography, preparative high performance liquid chromatography or a combination of the above. The specific methods used can be found in the examples described herein. Of course, other similar isolation and purification means can also be used. They can be characterized using conventional methods, including physical constants and spectral data.

[0172] Preparation Example

[0173] Preparation Example 1: Preparation of PE-1

[0174] Step 1: Preparation of tert-butyl 2-((4R,6R)-6-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethyl)-2,2-dimethyl-1,3-dioxan-4-yl)acetate

[0175] To a solution of tert-butyl 2-((4R,6R)-6-(2-aminoethyl)-2,2-dimethyl-1,3-dioxan-4-yl)acetate (500 mg, 1.83 mol, 1.0 eq), 2,5-dioxocyclopentyl (9H-fluoren-9-yl) carbonate (615.17 mg, 1.83 mmol, 1.0 eq) in tetrahydrofuran (6 ml) at 0 °C, sodium bicarbonate (307 mg, 3.66 mmol, 2 eq) was added (dissolved in 3 ml water). After the addition, the reaction was allowed to react at 25 °C for 16 hours. LCMS showed the reaction was completed. The reaction solution was diluted with water and extracted with ethyl acetate for 3 times. The combined organic phase was washed with brine once, dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the yellow solid compound tert-butyl 2-((4R,6R)-6-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethyl)-2,2-dimethyl-1,3-dioxan-4-yl)acetate (988 mg, crude without yield).

[0176] Step 2: Preparation of (3R,5R)-7-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3,5-dihydroxyheptanoic acid

[0177] To a solution of compound tert-butyl acetate 2-((4R,6R)-6-(2-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)ethyl)-2,2-dimethyl-1,3-dioxan-4-yl)(111 mg, 90%, 0.20 mmol, 1 eq) in tetrahydrofuran (10 mL) was added 10% hydrochloric acid (2 mL). After addition, the reaction was stirred at 25 °C for 2 hours, LCMS showed about 10% of product, the reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer ISCO, model ISCO-R2. The chromatographic column was YMC-Triart Prep C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% FA, formic acid) - acetonitrile, acetonitrile elution ratio from 39% to 69% elution) to give compound (3R,5R)-7-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3,5- dihydroxyheptanoic acid (17 mg, yield 21%).

[0178] LCMS (ESI): m / z, 422 [M+Na] + Rt = 1.662 min. 1 H NMR (400 MHz, Chloroform-d) δ 7.77 (d, J = 7.6 Hz, 2H), 7.59 (d, J = 8.0 Hz, 2H), 7.41 (t, J = 7.4 Hz, 2H), 7.36 - 7.29 (m, 2H), 5.09 (s, 2H), 4.48 (d, J = 6.6 Hz, 2H), 4.34 - 4.14 (m, 2H), 3.88 (t, J = 10.2 Hz, 1H), 3.63 - 3.47 (m, 1H), 3.17 - 3.06 (m, 1H), 2.52 - 2.55 (m, 3H).

[0179] Step 3: Preparation of (9H-fluoren-9-yl)methyl ((3R,5R)-3,5-dihydroxy-7-(((S)-2- hydroxy-3-((2S,5S,8S,11S,13R,15R,16aS,18R,19R,19aS,23R,24aS,25S,26aR,27S,28R,29aR,29bS)-19-methoxy-13-methyl-7,14-dimethylene-21-oxooctahydro-25H-2,27:5,8,15- trioxa-23,25-ethanol-2,28-methoxy[3,2',3':5,6]pyrano[4]tricyclo[6.2.2.0]dodecan-8,11- diyl)oxymethyl)oxymethyl)benzoate

[0180] A solution of compound (3R,5R)-7-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)- 3,5-dihydroxyheptanoic acid (2.9 mg, 7.26 μmol, 1.2 eq), eribulin mesylate (5 mg, 6.05 μmol, 1 eq), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (HATU, 2.76 mg, 7.26 μmol, 1.2 eq), N,N-diisopropylethylamine (2 μL, 12.11 μmol, 2 eq) in N,N-dimethylformamide (1 mL) was stirred in an ice bath for 5 min, then stirred at room temperature for 2 h. LCMS showed the starting material was consumed completely. The oil pump was used to concentrate to get (9H-fluoren-9-yl)methyl ((3R,5R)-3,5-dihydroxy-7-(((S)-2-hydroxy-3-((2S,5S,8S,11S,13R,15R,16aS,18R,19R,19aS,23R,24aS,25S,26aR,27S,28R,29aR,29bS)-19-methoxy-13-methyl-7,14-dimethylene-21-oxaoctahydro-25H-2,27:5,8,15-trioxy-23,25-ethanol-2,28-methoxy[3,2',3':5,6]pyrano[4](10.4 mg, crude without yield).

[0181] LCMS (ESI): m / z, 1133 [M+Na] + .

[0182] Step 4: (3R,5R)-7-amino-3,5-dihydroxy-N-((S)-2-hydroxy-3-((2S,5S,8S,11S,13R,15R,16aS,18R,19R,19aS,23R,24aS,25S,26aR,27S,28R,29aR,29bS)-19-methoxy-13-methyl-7,14-dimethylene-21-oxaoctahydro-25H-2,27,5,8,15-trioxy-23,25-ethoxy-2,28-methoxy[3,2-i]furo[2',3':5,6]pyrano[4,3-b][1,4]dioxepin-18-yl)propyl)heptanamide (PE-1)

[0183] To a solution of compound (9H-fluoren-9-yl)methyl ((3R,5R)-3,5-dihydroxy-7-(((S)-2-hydroxy-3-((2S,5S,8S,11S,13R,15R,16aS,18R,19R,19aS,23R,24aS,25S,26aR,27S,28R,29aR,29bS)-19-methoxy-13-methyl-7,14-dimethylene-21-oxaoctahydro-25H-2,27:5,8,15-trioxy-23,25-ethanol-2,28-methoxy[3,2',3':5,6]pyrano[4](10.4 mg, crude) in N,N-dimethylformamide (1 mL) was added 1,8-diazobicyclo[5.4.0]undec-7-ene (1.75 μL, 11.69 μmol, 3 eq) and stirred in ice bath for 5 min, then stirred at room temperature for 2 h, LCMS showed the starting material was consumed completely, purified by preparative HPLC (preparative HPLC, SHIMADZU, LC-20AP. Column: GS-120-10-C18AP. Mobile phase: water (0.225% HCOOH) - acetonitrile, water elution ratio from 10% to 45% elution) to give (3R,5R)-7-amino-3,5-dihydroxy-N-((S)-2-hydroxy-3-((2S,5S,8S,11S,13R,15R,16aS,18R,19R,19aS,23R,24aS,25S,26aR,27S,28R,29aR,29bS)-19-methoxy-13-methyl-7,14-dimethylene-21-oxaoctahydro-25H-2,27,5,8,15-trioxy-23,25-ethoxy-2,28-methoxy[3,2'-]furo[2',3':5,6]pyrano[4,3-b][1,4]dioxepin-18-yl)propyl)heptanamide (1.05 mg, yield 24.2%).

[0184] LCMS (ESI): m / z, 889 [M+H] + .

[0185] 1H NMR (400 MHz, DMSO-d6) δ 8.40 (s, 1H), 7.80 (s, 1H), 5.08-4.97 (m, 2H), 4.79 (d, J = 30.8 Hz, 2H), 4.64 (dt, J = 4.6, 2.6 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (td, J = 10.2, 4.2 Hz, 1H), 4.10 (d, J = 4.4 Hz, 2H), 4.02 (s, 1H), 3.93 (d, J = 5.6 Hz, 1H), 3.84-3.67 (m, 4H), 3.52 (d, J = 10.0 Hz, 4H), 3.05 (q, J = 6.8 Hz, 4H), 2.86-2.75 (m, 4H), 2.28-2.20 (m, 6H), 2.13 (t, J = 11.6 Hz, 3H), 2.06-1.82 (m, 10H), 1.74-1.57 (m, 8H), 1.55-1.39 (m, 6H), 1.36-1.14 (m, 6H), 1.04 (d, J = 6.4 Hz, 3H).

[0186] Preparation Example 2: Preparation of compound PE-2

[0187] Step 1: Preparation of compound PE-2-c

[0188] A solution of compound eribulin mesylate (8 mg, 9.69 μmol, 1 eq), (((9H-fluoren-9-yl)methoxy)carbonyl)-L-serine (3.8 mg, 11.69 μmol, 1.2 eq), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl)urea hexafluorophosphate (4.42 mg, 11.62 μmol, 1.2 eq), N,N-diisopropylethylamine (3.2 μL, 19.37 μmol, 2 eq) in N,N-dimethylformamide (1 mL) was stirred in ice bath for 5 min, then stirred at room temperature for 2 h. LCMS showed the starting material was consumed completely. The oil pump was used to concentrate the solution to get the crude compound PE-2-c (19.4 mg, crude yield not calculated).

[0189] LCMS (ESI): m / z, 1039 [M+H] + .

[0190] Step 2: Preparation of compound PE-2

[0191] To a solution of compound PE-2-c (crude, 19.4 mg) in N,N-dimethylformamide (1 mL) was added 1,8-diazobicyclo[5.4.0]undec-7-ene (8.37 μL) under ice-bath, and stirred for 5 min under ice-bath, then stirred for 2 h at room temperature. LCMS showed the starting material was consumed completely, and purified by reverse phase preparative HPLC (preparative HPLC, SHIMADZU, LC-20AP. Column: GS-120-10-C18AP. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution from 30% to 60%) to give compound PE-2 (3.11 mg, yield 39.4%, white solid).

[0192] LCMS (ESI): m / z, 817 [M+H] + .

[0193] 1 H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 7.92 (s, 1H), 5.03 (dd, J = 21.6, 2.6 Hz, 2H), 4.84 - 4.74 (m, 2H), 4.63 (dt, J = 4.2, 2.4 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (td, J = 10.2, 4.2 Hz, 1H), 4.10 (q, J = 4.8, 4.0 Hz, 3H), 4.06 - 3.99 (m, 1H), 3.83 - 3.67 (m, 4H), 3.58 - 3.49 (m, 6H), 3.19 - 2.99 (m, 6H), 2.84 (dd, J = 9.4, 1.8 Hz, 1H), 2.80 - 2.70 (m, 2H), 2.33 - 2.17 (m, 6H), 2.17 - 2.07 (m, 2H), 2.05 - 1.95 (m, 2H), 1.92 (q, J = 6.4, 5.8 Hz, 4H), 1.75 - 1.58 (m, 6H), 1.48 (dtd, J = 19.8, 11.2, 9.8, 5.2 Hz, 3H), 1.31 (ddt, J = 15.8, 11.4, 5.0 Hz, 3H), 1.26 - 1.10 (m, 2H), 1.06 - 0.98 (m, 3H).

[0194] Preparation Example 3: Preparation of compound PE-E2H

[0195] Step 1: Preparation of compound PE-E2H-a

[0196] To a solution of compound PE-E2K-a (synthesis of this compound please refer to the preparation of PE-E2K) (10 mg, 10.50 pmol, 1 eq) in MeOH (2 mL) was added wet Pd / C (10 mg), the reaction was stirred at 25 °C under hydrogen for 3 hours. LCMS showed the reaction was completed. The mixture was filtered through celite and concentrated, the product PE-E2H-a (7 mg, yield 69.85%) was obtained as a white solid by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution) LCMS (ESI): m / z, 954.4 [M+H] + .

[0197] Step 2: Preparation of compound PE-E2H

[0198] To a solution of compound PE-E2H-a (7 mg, 7.34 pmol, 1 eq) in DMF (2 mL) was added Et2NH (5.42 mg, 73.36 pmol, 10 eq), the reaction was stirred at 25 °C for 2 hours, LCMS showed the starting material was completely reacted. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to obtain white solid PE-E2H (3.70 mg, yield 52.03%), LCMS (ESI): m / z, 732.4 [M+H] + .

[0199] 1H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 2H), 5.58 (d, J = 8.0 Hz, 3H), 4.81 (s, 1H), 4.74 (s, 1H), 4.62 (t, J = 4.4 Hz, 1H), 4.56 (t, J = 4.4 Hz, 1H), 4.26 - 4.15 (m, 2H), 4.14 - 4.07 (m, 3H), 3.89 - 3.79 (m, 3H), 3.75 - 3.68 (m, 2H), 3.56 (s, 4H), 3.27 (d, J = 3.2 Hz, 6H), 2.89 - 2.81 (m, 2H), 2.70 (d, J = 7.2 Hz, 1H), 2.26 - 2.14 (m, 4H), 2.05 - 1.84 (m, 7H), 1.73 - 1.61 (m, 6H), 1.51 - 1.44 (m, 2H), 1.30 - 1.20 (m, 4H), 1.11 - 1.01 (m, 5H), 0.96 - 0.86 (m, 3H).

[0200] Preparation Example 4: Preparation of compound PE-E2K

[0201] Step 1: Preparation of compound PE-E2K-a

[0202] To a solution of compound eribulin mesylate (30 mg, 36.32 μmol, 1 eq) in THF (2 mL) was added NaHCO3(6.13 mg, 72.64 μmol, 2 eq) and Fmoc-OSu (fluorenylmethoxycarbonylsuccinimide, 13.75 mg, 39.95 μmol, 1.1 eq) at 0 °C, and the reaction was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The mixture residue was purified by silica gel column chromatography (C18, ISCO, R-40 g SepaFlash Silica Flash Column, Eluent of 10-75% water / CH3CN, 100 mL / min) to give the product PE-E2K-a (30 mg, yield 86.75%) as colorless oil. LCMS (ESI): m / z, 952.6 [M+H] + .

[0203] Step 2: Preparation of compound PE-E2K-b

[0204] To a solution of compound PE-E2K-a (15 mg, 15.75 pmol, 1 eq) in THF (tetrahydrofuran, 3 mL) and water (1 mL) was added potassium osmate dihydrate (490.97 pg, 1.58 pmol, 0.1 eq) and NaIO4(67.73 mg, 315.08 pmol, 20 eq) at 0 °C. The reaction was stirred at 25 °C for 2 h. To the reaction was added NaIO4(67.73 mg, 315.08 pmol, 20 eq) and the reaction was stirred at room temperature overnight. LCMS showed the reaction was complete. The mixture (and previous batch: 5 mg, small test) was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Synergi Max-RP 200 x 30 mm x 10 pm. Mobile phase: water (0.1% TFA) - acetonitrile, acetonitrile elution ratio from 40% to 70% elution) to give the product PE-E2K-b (16.60 mg, yield 82.68%) as a white solid. LCMS (ESI): m / z, 956.4 [M+H] + .

[0205] Step 3: Preparation of compound PE-E2K

[0206] To a solution of compound PE-E2K-b (16.6 mg, 17.36 pmol, 1 eq) in DMF (2 mL) was added Et2NH (6.41 mg, 86.81 pmol, 5 eq) and the reaction was stirred at 25 °C for 1 h. LCMS showed the starting material was consumed. The mixture was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 250 x 30 mm x 10 pm. Mobile phase: water (0.1% TFA) - acetonitrile, acetonitrile elution ratio from 10% to 40% elution) to give PE-E2K (7.50 mg, yield 58.86%) as a white solid.

[0207] LCMS (ESI): m / z, 734.3 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 7.67 (s, 2H), 5.23 (s, 1H), 4.70-4.53 (m, 2H), 4.46-4.28 (m, 2H), 4.19-4.05 (m, 3H), 3.94-3.67 (m, 5H), 3.61-3.47 (m, 1H), 3.33-3.21 (m, 4H), 3.01-2.70 (m, 4H), 2.65-2.52 (m, 2H), 2.41-2.34 (m, 2H), 2.30-1.87 (m, 8H), 1.85-1.10 (m, 14H), 0.97-0.90 (m, 2H).

[0208] Preparation Example 5: Preparation of compounds PE-E4HB1 and PE-E4HS1

[0209] Step 1: Preparation of compounds PE-E4H-a and PE-E4H-b

[0210] To a solution of compound PE-E2K-a (synthesis of this compound please refer to Preparation of PE-E2K) (10 mg, 10.50 pmol, 1 eq) in MeOH (2 mL) was added wet Pd / C (10 mg), the reaction was stirred at 25 °C under hydrogen overnight. LCMS showed the reaction was completed. The mixture was filtered through celite and rotary evaporated to dryness, the product PE-E4H-a (7 mg, yield 69.7%) was obtained as a white solid by high performance liquid chromatography preparation (Prep-HPLC, Oriendo, BRIX-2860. Column: YMC-Triart Prep C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 60% to 90% elution). + ; and PE-E4H-b (2.50 mg, yield 24.89%), LCMS (ESI): m / z, 954.4 [M+H] + .

[0211] Step 2: Preparation of compound PE-E4HB1

[0212] To a solution of compound PE-E4H-a (7 mg, 7.32 pmol, 1 eq) in DMF (2 mL) was added Et2NH (5.41 mg, 73.21 pmol, 10 eq), the reaction was stirred at 25 °C for 2 h, LCMS showed the starting material was consumed completely. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% FA) - acetonitrile, the elution ratio of acetonitrile was eluted from 20% to 50%), to give white solid PE-E4HB1 (4.90 mg, yield 40.52%, dimethyl sulfonate), LCMS (ESI): m / z, 734.4 [M+H] + .

[0213] 1 H NMR (400 MHz, DMSO-d6) d 8.41 (s, 6H), 5.42 (s, 1H), 5.26 (s, 1H), 4.95 (t, J = 9.2 Hz, 1H), 4.62 (s, 1H), 4.11 (s, 2H), 3.65 (t, J = 9.0 Hz, 3H), 3.28 (d, J = 3.0 Hz, 3H), 2.79 (s, 14H), 2.56 (d, J = 15.9 Hz, 1H), 2.29 (s, 3H), 1.99 - 1.70 (m, 4H), 1.47 (s, 3H), 1.33 - 1.21 (m, 2H), 1.12 (t, J = 7.2 Hz, 19H), 1.07 (d, J = 10.4 Hz, 1H), 0.96 - 0.72 (m, 4H).

[0214] Step 3: Preparation of compound PE-E4HS1

[0215] To a solution of compound PE-E4H-b (2.50 mg, 2.61 pmol, 1 eq) in DMF (2 mL) was added Et2NH (1.93 mg, 26.15 pmol, 10 eq), the reaction was stirred at 25 °C for 2 h, LCMS showed the starting material was consumed completely. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% FA) - acetonitrile, the elution ratio of acetonitrile was eluted from 20% to 50%), to give white solid PE-E4HS1 (1.90 mg, yield 99.01%), LCMS (ESI): m / z, 734.4 [M+H] + .

[0216] Preparation Example 6: Preparation of compound PE-5

[0217] Step 1: Preparation of (1R,2S,5S)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2- carboxylic acid

[0218] To a solution of (1R,2S,5S)-methyl 6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2- carboxylate hydrochloride (200 mg, 972.37 pmol, 1 eq) in THF (4 mL) and H2O (4 mL) was added LiOH.H2O (163.36 mg, 108.18 pL, 3.89 mmol, 4 eq), and the mixture was stirred at 25 °C for 16 h. The reaction was monitored by LCMS, the starting material was consumed completely and the main product was generated, the pH was adjusted to 3-4 with HC1, to give 6,6-dimethyl-3-(1R,2S,5S)-azabicyclo[3.1.0]hexane-2-carboxylic acid crude, which was used directly in the next step (140 mg, crude without yield). LCMS (ESI): m / z, 156 [M+H] + .

[0219] Step 2: Preparation of (1R,2S,5S)-3-((9H-fluoren-9-yl)methoxy)carbonyl)-6,6- dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid

[0220] To a solution of 6,6-dimethyl-3-(1R,2S,5S)-azabicyclo[3.1.0]hexan-2- carboxylic acid (140 mg, 902.06 umol, 1 eq) in NaHC03(1 mL) (adjust pH > 7), Fmoc-OSu (304.29 mg, 217.35 uL, 902.06 umol, 1 eq) was added and the mixture was stirred at 25 °C for 5 h. The reaction was monitored by LCMS, the starting material was consumed completely and the main product was generated. The product was extracted with EA (ethyl acetate, 5 mL x 3) and H20 (5 mL x 2), then the organic phase was combined. The water phase was adjusted to pH 3-4 with HC1, then extracted with EA (10 mL x 3), and the organic phase was combined. The solvent was removed under reduced pressure, and the product was purified by preparative HPLC (Oriendo, Lab311-DJ-R7. Column: Welch Xtimate C18 250 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 39% to 69% elution) and lyophilized to give the product (1R,2S,5S)-3-((9H-fluoren-9-yl)methoxy)carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid (80 mg, 23.50% yield) as a white solid. LCMS (ESI): m / z, 400 [M+Na] + .

[0221] Step 3: Preparation of compound PE-5d

[0222] To a solution of eribulin mesylate (9.67 mg, 13.25 umol, 1 eq) and (1R,2S,5S)-3-(((9H-fluoren-9-yl)methoxy)carbonyl)-6,6-dimethyl-3-azabicyclo[3.1.0]hexane-2-carboxylic acid (5 mg, 13.25 umol, 1 eq) in DMF (5 mL), HATU (6.55 mg, 17.22 umol, 1.3 eq) and DIPEA (N,N-diisopropylethylamine, 5.14 mg, 6.57 uL, 39.75 umol, 3 eq) were added at 0 °C and the mixture was stirred at 25 °C for 16 h. The reaction was monitored by LCMS, the starting material was consumed completely and the main product was generated. The product was purified by preparative HPLC (ISCO, Lab311-DJ-R5. Column: Phenomenex Luna C18 250 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 57% to 87% elution) and lyophilized to give compound PE-5d (12.50 mg, 86.61% yield) as a white solid. LCMS (ESI): m / z, 1011 [M+Na] + .

[0223] Step 4: Preparation of compound PE-5

[0224] To compound PE-5d (10.50 mg, 9.64 pmol, 1 eq) in DMF (2 mL) was added DBU (1,8-diazabicyclo[5.4.0]undec-7-ene, 146.74 pg, 1.44 eq), the mixture was stirred at 25 °C for 1 h. LCMS monitored the reaction, the starting material was consumed completely and the main product was formed, preparative purification was performed using high performance liquid chromatography (preparative chromatography manufacturer ISCO, model Lab311-DJ-R5. The column was Phenomenex Luna C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution), lyophilized to get compound PE-5 (1 mg, yield 11.96%, purity: 100%) as a white solid. -1

[0225] LCMS (ESI): m / z, 867 [M+H] + . 1 ​H NMR (400 MHz, DMSO-d6) δ 8.25 (s, 1H), 8.02 (t, J = 5.9 Hz, 1H), 5.03 (d, J = 21.3 Hz, 2H), 4.83 (s, 1H), 4.76 (s, 2H), 4.64 (t, J = 4.0 Hz, 1H), 4.55 (t, J = 4.3 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (td, J = 10.2, 4.1 Hz, 1H), 4.13 - 4.06 (m, 3H), 4.01 (d, J = 9.4 Hz, 1H), 3.77 (ddd, J = 13.5, 10.1, 6.7 Hz, 2H), 3.69 (ddd, J = 9.3, 5.6, 2.9 Hz, 1H), 3.52 (q, J = 11.1, 9.9 Hz, 6H), 3.26 (s, 6H), 3.12 (dt, J = 11.4, 5.4 Hz, 2H), 3.04 (dt, J = 10.4, 6.2 Hz, 2H), 2.89 - 2.81 (m, 2H), 2.61 - 2.55 (m, 1H), 2.33 (d, J = 14.7 Hz, 2H), 2.14 (d, J = 12.0 Hz, 1H), 2.07 - 1.96 (m, 2H), 1.92 (q, J = 6.2, 5.7 Hz, 4H), 1.66 (tq, J = 13.4, 9.2, 6.5 Hz, 6H), 1.56 - 1.44 (m, 2H), 1.41 (d, J = 7.1 Hz, 1H), 1.38 - 1.26 (m, 3H), 1.24 - 1.14 (m, 2H), 1.03 (d, J = 6.5 Hz, 3H), 0.98 (s, 6H).

[0226] Preparation Example 7: Preparation of compound PE-7

[0227] Step 1: Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-((S)-2- oxopyrrolidin-3-yl)propanoic acid

[0228] Compound (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-((S)-3- oxopyrrolidin-3-yl)propanoate (300 mg, 1.0 eq) was dissolved in a mixture solution of tetrahydrofuran (3 mL) and water (3 mL), after the addition of lithium hydroxide monohydrate (132 mg, 3.0 eq) and stirred at 25 °C for 3 hours. LCMS detection showed that the reaction was complete. After treatment, the reaction was diluted with pure water (10 mL), neutralized to pH = 5 with 1M dilute hydrochloric acid, extracted twice with a mixture of dichloromethane and methanol (volume ratio 10:1), the organic phase was combined, washed with saturated brine, and the organic phase was dried and concentrated to give the crude product (S)-2-((tert-butoxycarbonyl)amino)-3-((S)-2- oxopyrrolidin-3-yl)propanoic acid (350 mg) which was used directly in the next step.

[0229] LCMS: m / z = 295.2 [M+Na] + ; Rt = 0.761 min.

[0230] Step 2: Preparation of compound PE-7-d

[0231] Compound eribulin mesylate (10 mg, 1.0 eq, purchased from Haoyuan) and (S)-2-((tert-butoxycarbonyl)amino)-3-((S)-2-oxopyrrolidin-3-yl)propanoic acid (3.73 mg, 1.0 eq) were dissolved in DMF (2 mL), after the addition of DIPEA (6.79 μL, 3.0 eq) and HATU (6.77 mg, 1.3 eq) and stirred at 25 °C for 16 hours, LCMS detection showed that the starting material was completely reacted. After treatment, the reaction was filtered, and the filtrate was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 30% to 60% elution) to give white solid compound PE-7-d (10 mg, yield 74%).

[0232] LCMS: m / z = 984.5 [M+H] + ; Rt = 2.040 min.

[0233] Step 3: Preparation of compound PE-7

[0234] Compound PE-7-d (10 mg, 1.0 eq) was dissolved in anhydrous dichloromethane (2 mL), after cooling in ice bath, trifluoroacetic acid (280 μL) was added, then the reaction was stirred at 25 °C for 16 hours. LCMS detection showed the starting material was consumed completely, the reaction was concentrated to dryness, the residue was dissolved in DMF and then purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 50 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give white solid compound PE-7 (5.68 mg, yield 79%). LCMS: m / z = 884.5 [M+H] + ; Rt = 2.040 min.

[0235] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.05 (t, J = 5.9 Hz, 1H), 7.66 (s, 2H), 6.20 (d, J = 3.0 Hz, 1H), 5.95 (d, J = 3.0 Hz, 1H), 4.97 - 4.83 (m, 3H), 4.81 (s, 2H), 4.75 (s, 3H), 4.67 - 4.63 (m, 2H), 4.32 (dd, J = 19.2, 7.8 Hz, 3H), 4.06 (q, J = 6.8, 6.3 Hz, 1H), 4.01 (d, J = 3.0 Hz, 1H), 3.88 - 3.75 (m, 3H), 3.70 (dd, J = 8.4, 3.9 Hz, 3H), 3.61 (d, J = 4.6 Hz, 1H), 3.21 (s, 4H), 3.17 - 3.07 (m, 4H), 2.98 (dd, J = 9.3, 2.1 Hz, 1H), 2.76 (dt, J = 14.5, 5.7 Hz, 3H), 2.35 (s, 1H), 2.31 - 2.16 (m, 3H), 2.13 (td, J = 15.9, 15.4, 6.8 Hz, 1H), 1.96 (dd, J = 5.8, 3.1 Hz, 1H), 1.96 - 1.89 (m, 1H), 1.89 - 1.81 (m, 2H), 1.76 (dt, J = 8.4, 4.2 Hz, 2H), 1.74 - 1.66 (m, 7H), 1.69 - 1.61 (m, 1H), 1.60 (d, J = 8.2 Hz, 3H), 1.59 - 1.43 (m, 1H), 1.42 (dd, J = 11.1, 5.1 Hz, 2H), 1.31 (s, 2H), 1.04 (d, J = 6.3 Hz, 4H), 0.94 (q, J = 12.0 Hz, 1H).

[0236] Preparation Example 8: Preparation of compound PE-10

[0237] Step 1: Preparation of compound PE-10b

[0238] To a solution of eribulin mesylate (10 mg, 12.1 μmol, 1 eq) and bromoethanol (2.27 mg, 18.2 μmol, 1.5 eq) in N,N-dimethylformamide (1.5 mL) was added potassium carbonate (5.02 mg, 36.3 μmol, 3 eq) and placed in a 25 °C oil bath for 4 hours, LCMS showed product formation. After filtration, PE-10b (7.8 mg, yield 73.5%) was obtained using high performance liquid chromatography preparation (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution). LCMS (ESI): m / z, 774 [M+H] + .

[0239] Step 2: Preparation of compound PE-10c

[0240] To a solution of compound PE-10b (7.8 mg, 10.08 μmol, 1 eq) and (3R,5R)-7-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3,5-dihydroxyheptanoic acid (4.83 mg, 12.09 μmol, 1.2 eq, the synthesis steps of this intermediate refer to Preparation Example of PE-1) in N,N-dimethylformamide (2 mL) was added N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (5.75 mg, 15.12 μmol, 1.5 eq) and N,N-diisopropylethylamine (3.33 μL, 20.16 μmol, 2 eq) and stirred at room temperature for 2 hours, LCMS showed the starting material was completely reacted, and the crude product (22.5 mg, crude yield not calculated) was obtained by oil pump dryness. + .

[0241] Step 3: Preparation of compound PE-10

[0242] To a solution of compound PE-10c (22.5 mg, purity 51.7%, 10.08 pmol, 1 eq) in N,N-dimethylformamide (2 mL) was added 1,8-diazobicyclo[5.4.0]undec-7-ene (4.5 pL, 30.24 pmol, 3 eq) and stirred at room temperature for 1 h, LCMS showed the starting material was consumed completely, after filtration, the white solid compound PE-10 (7.8 mg, yield 73.5%) was obtained by using high performance liquid chromatography preparation (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The column was GS-120-10-C18AP. The mobile phase was water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 20% to 50% elution).

[0243] LCMS (ESI): m / z, 933 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) d 8.42 (s, 1H), 7.81 (s, 1H), 5.06 (s, 1H), 5.00 (s, 1H), 4.84 (s, 1H), 4.76 (s, 1H), 4.64 (s, 1H), 4.56 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.4 Hz, 1H), 4.19 (d, J = 13.2 Hz, 1H), 4.10 (d, J = 4.8 Hz, 3H), 4.06 - 3.93 (m, 2H), 3.85 - 3.64 (m, 5H), 3.51 (s, 6H), 3.27 (d, J = 7.2 Hz, 8H), 3.06 (s, 2H), 2.87 - 2.70 (m, 5H), 2.58 (d, J = 16.2 Hz, 1H), 2.34 (s, 1H), 2.24 (dd, J = 16.8, 9.8 Hz, 5H), 2.14 (d, J = 12.0 Hz, 2H), 2.06 - 1.96 (m, 2H), 1.93 (s, 4H), 1.67 (t, J = 12.2 Hz, 6H), 1.50 (s, 6H), 1.35 (d, J = 18.2 Hz, 3H), 1.26 - 1.12 (m, 2H), 1.04 (d, J = 6.4 Hz, 3H).

[0244] Preparation Example 9: Preparation of compound PE-12

[0245] Step 1: Preparation of compound PE-12-a

[0246] To a solution of eribulin mesylate (5 mg, 6.05 μmol, 1 eq) in dichloromethane (1 mL) was added N, N-diisopropylethylamine (1.5 μL, 9.08 μmol, 1.5 eq) and N-BOC p-aminobenzenesulfonyl chloride (1.94 mg, 6.66 μmol, 1.1 eq) under ice bath, and stirred at room temperature for 2 hours. LCMS showed the starting material was consumed completely. The crude product PE-12-a (13 mg, crude, 9.6 mg in theory) was obtained by oil pump concentration.

[0247] LCMS (ESI): m / z, 1007 [M+Na] + .

[0248] Step 2: Preparation of compound PE-12

[0249] To a solution of compound PE-12-a (7.68 mg, 4.84 μmol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.1 mL) under ice bath, and stirred at room temperature for 2 hours. LCMS showed the starting material was consumed completely. The reaction solution was concentrated, then dissolved in N, N-dimethylformamide and filtered. The filtrate was purified by high performance liquid chromatography (Oriendo, BRIX-2860. Column: Synergi Max-RP 250 x 40 mm x 10 μm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give compound PE-12 (0.52 mg, yield 12.14%) as a white solid.

[0250] LCMS (ESI): m / z, 907 [M+Na] + .

[0251] Preparation Example 10: Preparation of compound PE-16

[0252] Step 1: Preparation of compound PE-16-a

[0253] To a solution of compound eribulin mesylate (5 mg, 6.05 pmol, 1 eq) and compound 6-((tert-butoxycarbonyl)amino)spiro[3.3]heptane-2-carboxylic acid (1.85 mg, 7.26 pmol, 1.2 eq) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (2 pL, 12.11 pmol, 2 eq) and N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl)urea hexafluorophosphate (2.76 mg, 7.26 pmol, 1.2 eq) under ice bath, and stirred at room temperature for 2 hours, LCMS showed the starting material was consumed. The product PE-16-a (4.60 mg, yield 78.5%) was obtained as a white solid by high performance liquid chromatography preparation (Preparative Chromatography manufacturer Oriendo, model BRIX-2860. The column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 40% to 70% elution).

[0254] LCMS (ESI): m / z, 967 [M+H] + .

[0255] Step 2: Preparation of compound PE-16

[0256] To a solution of compound PE-16-a (4.60 mg, 4.76 pmol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.1 mL) under ice bath, and then stirred at room temperature for 2 hours, LCMS showed the starting material was consumed, the residue was concentrated by oil pump, and the product PE-16 (1.66 mg, yield 39.3%) was obtained as a white solid by high performance liquid chromatography preparation (Preparative Chromatography manufacturer Oriendo, model BRIX-2860. The column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution).

[0257] LCMS (ESI): m / z, 867 [M+H] + .

[0258] Preparation Example 11: Preparation of compound PE-18

[0259] Step 1: Preparation of compound PE-18-a

[0260] To a solution of eribulin mesylate (10 mg, 12.11 pmol, 1 eq) in N,N- dimethylformamide (1 mL) was added N,N-diisopropylethylamine (4.69 mg, 6 pL, 36.32 pmol, 3 eq), 2-(7-azabenzotriazol-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5.52 mg, 14.53 pmol, 1.2 eq) and 4-((tert- butoxycarbonyl)amino)bicyclo[2.2.1]heptane-1 -carboxylic acid (2.55 mg, 10 pmol, 1 eq) successively at room temperature. The mixture was stirred at 20 °C for 3 h. LCMS showed the starting material was consumed completely. The reaction solution was purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6, 7). The column was Phenomenex Luna C18 250 x 50 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution). The white solid product PE-18-a (6 mg, 6.20 pmol, 51% yield) was obtained after lyophilization.

[0261] LCMS (ESI): m / z, 967.6 [M+H] + .

[0262] Step 2: Preparation of compound PE-18

[0263] PE-18-a (6 mg, 6.20 pmol, 1 eq) was added to dichloromethane (1 mL) and trifluoroacetic acid (0.1 mL) at room temperature. The mixture was stirred at 20 °C for 1 h. LCMS showed the reaction was complete. The reaction solution was purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6, 7). The column was Phenomenex Luna C18 250 x 50 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution). The white solid product PE-18 (3.09 mg, 3.38 pmol, 55% yield) was obtained after lyophilization.

[0264] LCMS (ESI): m / z, 867.6 [M+H] + .

[0265] Preparation Example 12: Preparation of compound PE-21A

[0266] Step 1: Preparation of compound 2-[(tert-butyldimethylsilyl)oxy]ethyl 4- nitrophenyl carbonate

[0267] To a solution of compound tert-butyldimethylhydroxyethoxysilane (3 g, 17.01 mmol, 1 eq) and compound bis(p-nitrophenyl) carbonate (10.35 g, 6.90 mL, 34.03 mmol, 2 eq) in DMF (20 mL), DIPEA (6.60 g, 8.44 mL, 51.04 mmol, 3 eq) was added, the reaction was stirred at 25 °C for 15 h, LCMS showed the starting material was consumed completely, extracted with dichloromethane (10 mL x 3), the combined organic phase was washed with water (20 mL x 2), saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate 5:1) to give compound 2-[(tert-butyldimethylsilyl)oxy]ethyl 4-nitrophenyl carbonate (4.52 g, yield 77.73%) as yellow oil. LCMS (ESI): m / z, 342 [M+H] + .

[0268] Step 2: Preparation of compound PE-21A-a

[0269] To a solution of compound 2-((tert-butyldimethylsilyl)oxy)ethyl (4-nitrophenyl) carbonate (2.25 mg, 6.58 pmol, 1.2 eq) and eribulin mesylate (4 mg, 5.48 pmol, 1 eq) in DMF (1 mL), DIPEA (N,N-diisopropylethylamine, 1.42 mg, 1.81 pL, 10.96 pmol, 2 eq) was added at 0 °C, after reaction at 25 °C for 15 h, TLC showed the starting material was consumed, dried directly for next step, compound PE-21A-a (5.11 mg, crude without yield) as brown oil, LCMS (ESI): m / z, 945 [M+H] + .

[0270] Step 3: Preparation of compound PE-21A

[0271] To a solution of PE-21A-a (5.11 mg, 5.48 pmol, 1 eq) in DCM (800 pL) was added TFA (625 pg, 80 pL, 5.48 pmol, 1 eq) at 0 °C, and the mixture was stirred at 0-25 °C for 2 h. LCMS showed the starting material was consumed. Purification by preparative high performance liquid chromatography (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60%) afforded compound PE-21A (600 pg, yield 13.38%, purity 100%) as a white solid. LCMS (ESI): m / z, 840 [M+Na] + .

[0272] Preparation Example 13: Preparation of compound PE-21B

[0273] Step 1: Preparation of compound (S)-1-((tert-butyldimethylsilyl)oxy)propan-2-ol

[0274] To a solution of compound (S)-propane-1,2-diol (1 g, 13.14 mmol, 1 eq) in DCM (10 mL) was added TBSCl (tert-butyldimethylsilyl chloride, 2.18 g, 2.50 mL, 14.46 mmol, 1.1 eq) and imidazole (894.73 mg, 813.39 pL, 13.14 mmol, 1 eq) at 0 °C. After addition, the mixture was stirred at 25 °C for 16 h. The reaction mixture was poured into ice aqueous sodium bicarbonate solution (60 mL), and then stirred for 2 min. The mixture was extracted with DCM (60 mL x 3). The organic phase was combined, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound (S)-1-((tert-butyldimethylsilyl)oxy)propan-2-ol (2.30 g, yield 91.94%) as colorless oil.

[0275] 1 H NMR (400 MHz, Chloroform-d) d 3.82-3.78 (m, 1H), 3.59-3.56 (m, 1H), 3.36-3.31 (m, 1H), 2.52 (br s, 1H), 1.11 (d, J = 8.0 Hz, 3H), 0.90 (s, 9H), 0.06 (s, 6H).

[0276] Step 2: Preparation of compound PE-21B-a

[0277] To a solution of (S)-1-((tert-butyldimethylsilyl)oxy)propan-2-ol (761.40 mg, 4 mmol, 1 eq) in DMF (10 mL) was added di(p-nitrophenyl) carbonate (2.43 g, 1.62 mL, 8 mmol, 2 eq) and DIPEA (1.55 g, 1.98 mL, 12 mmol, 3 eq) at 0 °C. The reaction was stirred at 25 °C for 16 h. LCMS showed the formation of new compound. The reaction was concentrated to dryness to give residue, which was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:30 to 1:15) to give PE-21B-a (900 mg, yield 63.30%) as light yellow oil.

[0278] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.32 (d, J = 8.0 Hz, 2H), 7.51 (d, J = 8.0 Hz, 2H), 4.88-4.86 (m, 1H), 3.78-3.66 (m, 2H), 1.28 (d, J = 8.0 Hz, 3H), 0.86 (s, 9H), 0.06 (s, 6H).

[0279] Step 3: Preparation of compound PE-21B-b

[0280] To a solution of compound PE-21B-a (2.43 mg, 6.85 μmol, 1 eq) in DMF (1 mL) was added eribulin mesylate (5.66 mg, 6.85 μmol, 1 eq) and DIPEA (2.66 mg, 3.40 μL, 20.55 μmol, 3 eq) and HOBt (1-hydroxybenzotriazole, 925.61 μg, 6.85 μmol, 1 eq) at 0 °C. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was complete, the reaction was concentrated to give crude product PE-21B-b (7 mg, crude) as yellow oil.

[0281] LCMS (ESI): m / z, 968.7 [M+Na] + , Rt = 2.46 min.

[0282] Step 4: Preparation of compound PE-21B

[0283] To a solution of compound PE-21B-b (6.48 mg, 6.85 pmol, 1 eq) in DCM (1 mL) was added trifluoroacetic acid (0.1 mL) at room temperature. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was complete. The reaction was concentrated, the residue was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30-60% elution) and lyophilized to give PE-21B (360 pg, yield 6.32%, purity 96.6%) as a white solid.

[0284] LCMS (ESI): m / z, 854.5 [M+Na] + Rt = 2.153 min. HPLC: Rt = 5.683 min.

[0285] Preparation Example 14: Preparation of compound PE-21C

[0286] Step 1: Preparation of compound PE-21C-a

[0287] To a solution of compound 2-((tert-butyldiphenylsilyl)oxy)-1-cyclopropylethan-1-ol (500 mg, 1.47 mmol, 1 eq, the synthesis of this compound please refer to the example of LA-31B) in DMF (20 mL) was added compound NPC (p-nitrophenyl chloroformate, 2.68 g, 8.81 mmol, 6 eq) and DIPEA (1.14 g, 8.81 mmol, 6 eq), the reaction was stirred at room temperature for 2 h. LCMS showed the reaction was complete. The reaction was rotary evaporated under reduced pressure, to the mixture was added DCM (50 mL) and water (100 mL), the mixture was extracted with DCM 3 times, 50 mL each time, the combined organic phase was washed with saturated brine (100 mL), dried with anhydrous Na2S04, filtered and rotary evaporated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether 0:1 to 1:5) to give the product PE-21C-a (426 mg, yield 57.38%) as a colorless oil.

[0288] 1H NMR (400 MHz, CDC13) δ 8.29 - 8.25 (m, 2H), 7.72 - 7.68 (m, 4H), 7.46 - 7.37 (m, 6H), 7.36 - 7.31 (m, 2H), 4.34 - 4.27 (m, 1H), 3.91 (d, J = 5.6 Hz, 2H), 1.09 - 1.05 (m, 10H), 0.64 - 0.49 (m, 3H), 0.29 - 0.21 (m, 1H).

[0289] Step 2: Preparation of compound PE-21C-b

[0290] To a solution of compound eribulin mesylate (10 mg, 12.11 μmol, 1 eq) and PE-21C-a (7.35 mg, 14.53 μmol, 1.2 eq) in DMF (2 mL), compound DIPEA (4.69 mg, 36.32 μmol, 3 eq) and HOBt (1.64 mg, 12.11 μmol, 1 eq) were added, and the reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was rotary evaporated under reduced pressure to give the product PE-21C-b (13.27 mg, crude) as colorless oil. LCMS (ESI): m / z, 1118.8 [M+Na] + Rt = 3.950 min.

[0291] Step 3: Preparation of compound PE-21C

[0292] To a solution of compound PE-21C-b (12.52 mg, crude) in DMF (2 mL), triethylamine trifluoride hydrochloride (0.04 mL) was added at 0 °C, and the reaction was stirred at 65 °C for 2.5 hours. LCMS showed the starting material was completely consumed. The mixture was prepared by high performance liquid chromatography (Prep HPLC, Oriendo, BRIX-2860-R6. The column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of water was from 30% to 60% elution) to give PE-21C (4.2 mg, yield 42.87%) as white solid. LCMS (ESI): m / z, 880.7 [M+Na] + Rt = 1.985 min.

[0293] 1H NMR (400 MHz, DMSO-d6) δ 6.93 - 6.79 (m, 1H), 5.10 - 4.96 (m, 2H), 4.87 - 4.49 (m, 5H), 4.33 - 3.98 (m, 6H), 3.86 - 3.64 (m, 3H), 3.57 - 3.42 (m, 4H), 3.26 (s, 6H), 3.02 - 2.54 (m, 7H), 2.35 - 1.82 (m, 12H), 1.78 - 0.86 (m, 17H), 0.51 - 0.18 (m, 4H).

[0294] Preparation Example 15: Preparation of compound PE-21D

[0295] Step 1: Preparation of compound PE-21D-a

[0296] To a solution of compound eribulin mesylate (10 mg, 12.11 μmol, 1 eq) and PY-4Car2-b (5.82 mg, 15.74 μmol, 1.3 eq) in THF (20 mL), compound DIPEA (4.69 mg, 36.32 μmol, 3 eq) and HOBt (1.64 mg, 12.11 μmol, 1 eq) were added, and the reaction was stirred at room temperature for 2.5 hours. LCMS showed that the reaction was completed. The reaction was rotary evaporated under reduced pressure to obtain the product PE-21D-a (11.60 mg, crude) as colorless oil. LCMS (ESI): m / z, 960.9 [M+H] + , Rt = 1.128 min.

[0297] Step 2: Preparation of compound PE-21D

[0298] To a solution of compound PE-21D-a (11.60 mg, crude) in DCM (2 mL), TFA (0.4 mL) was added at 0 °C, and the reaction was stirred at room temperature for 2 hours. LCMS showed that the starting material was completely reacted. The reaction was rotary evaporated under reduced pressure, and the mixture was prepared by high performance liquid chromatography (Prep HPLC, Oriendo, BRIX-2860. The column was Welch Xtimate C18 200 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) to obtain PE-21D (1.40 mg, yield 13.70%) as a white solid. LCMS (ESI): m / z, 808.4 [M+H] + , Rt = 2.128 min.

[0299] Preparation Example 16: Preparation of compound PE-21DA

[0300] Step 1: Preparation of compound 1-{[(4-{[(tert-butoxy)carbonyl]amino}butoxy- carbonyl]oxy}-4-nitrobenzene

[0301] To a solution of compound 4-(Boc-amino)-1-butanol (100 mg, 528.40 pmol, 1 eq) in DMF (5 mL) was added bis(4-nitrophenyl) carbonate (642.98 mg, 428.65 pL, 2.11 mmol, 4 eq) and DIPEA (409.74 mg, 523.97 pL, 3.17 mmol, 6 eq), 33 °C for 1 h, LCMS showed the reaction was completed, the residue was purified by silica gel column chromatography (C18, ISCO, R-120 g SepaFlash Silica Flash Column, Eluent of 10-60% water / CH3CN, 60 mL / min) to give compound 1-{[(4-{[(tert-butoxy)carbonyl]amino}butoxy- carbonyl]oxy}-4-nitrobenzene (131 mg, yield 69.96%) as a white solid.

[0302] LCMS (ESI): m / z, 255 [M-100+H] + . 1 H NMR (400 MHz, Chloroform-d) d 8.29 (d, J = 9.1 Hz, 2H), 7.39 (d, J = 9.2 Hz, 2H), 4.58 (s, 1H), 4.31 (t, J = 6.5 Hz, 2H), 3.19 (t, J = 6.7 Hz, 2H), 1.79 (dt, J = 8.8, 6.6 Hz, 2H), 1.66 (d, J = 7.3 Hz, 2H), 1.45 (s, 9H).

[0303] Step 2: Preparation of compound PE-21DA-a

[0304] To a solution of compound 1-{[(4-{[(tert-butoxy)carbonyl]amino}butoxy- carbonyl]oxy}-4-nitrobenzene (9.71 mg, 27.40 pmol, 2 eq) and eribulin mesylate (10 mg, 13.70 pmol, 1 eq) in THF (1 mL) was added DIPEA (5.31 mg, 6.79 pL, 41.10 pmol, 3 eq) and HOBT (1.85 mg, 1.23 pL, 13.70 pmol, 1 eq) at 0 °C, 35 °C for 1 h, LCMS showed the starting material was consumed, dried directly for next step. LCMS (ESI): m / z, 945 [M+H] + .

[0305] Step 3: Preparation of compound PE-21DA

[0306] To a solution of PE-21DA-a (12.95 mg, 13.70 pmol, 1 eq) in DCM (1 mL) was added TFA (614 mg, 400 pL, 5.39 mmol, 393.0275 eq) at 0 °C, and the reaction was stirred at 0 °C for 90 min. LCMS showed the starting material was consumed. The reaction was concentrated to dryness to give a crude product, which was purified by high performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was Phenomenex Luna C18 150 x 25 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 20% to 50% elution). The white solid compound PE-21DA (4 mg, yield 34.55%, purity 95%) was obtained after lyophilization.

[0307] LCMS (ESI): m / z, 845 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) d 8.42 (s, 1H), 6.97 (s, 1H), 6.20 (d, J = 3.0 Hz, 1H), 5.95 (d, J = 3.0 Hz, 1H), 4.95 (s, 1H), 4.87 (s, 1H), 4.81 (s, 1H), 4.75 (s, 1H), 4.65 (s, 1H), 4.36 - 4.27 (m, 2H), 4.07 - 3.99 (m, 2H), 3.93 (t, J = 6.4 Hz, 2H), 3.84 - 3.77 (m, 2H), 3.70 (s, 3H), 3.60 (d, J = 4.1 Hz, 3H), 3.52 (s, 5H), 3.21 (s, 5H), 3.02 - 2.91 (m, 4H), 2.78 (d, J = 6.8 Hz, 1H), 2.72 (q, J = 7.6, 6.7 Hz, 3H), 2.67 (q, J = 1.9 Hz, 1H), 2.33 (dt, J = 3.8, 1.9 Hz, 2H), 2.11 (dd, J = 15.8, 7.4 Hz, 1H), 1.90 (dd, J = 35.4, 11.5 Hz, 3H), 1.77 - 1.67 (m, 4H), 1.66 - 1.45 (m, 8H), 1.31 (t, J = 9.1 Hz, 2H), 1.04 (d, J = 6.4 Hz, 3H).

[0308] Preparation Example 17: Preparation of compound PE-21E

[0309] Step 1: Preparation of 5-[(tert-butyldimethylsilyl)oxy]-pentyl-4-nitrophenyl carbonate

[0310] 5-(tert-butyldimethylsilyl)-1-pentanol (218.41 mg, 1 mmol, 1 eq) was dissolved in DMF (3 mL), then di(p-nitrophenyl) carbonate (912.63 mg, 608.42 μL, 3 mmol, 3 eq) and DIPEA (387.72 mg, 495.81 μL, 3 mmol, 3 eq) were added. The reaction was reacted at 35 °C for 3 h. LCMS showed that the new compound was generated. The reaction was diluted with water (50 mL) and extracted with dichloromethane (50 mL x 3). The combined organic phase was washed with water (100 mL), dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (PE / EA (petroleum ether / ethyl acetate) = 93% / 7%) to give 5-[(tert-butyldimethylsilyl)oxy]-pentyl-4-nitrophenyl carbonate (330 mg, yield 86.05%) as a light yellow oil.

[0311] 1 H NMR (400 MHz, DMSO-d6) δ 8.39 - 8.25 (m, 2H), 7.60 - 7.45 (m, 2H), 4.24 (t, J = 6.4 Hz, 2H), 3.58 (t, J = 6.4 Hz, 2H), 1.77 - 1.59 (m, 2H), 1.60 - 1.45 (m, 2H), 1.46 - 1.33 (m, 2H), 0.85 (s, 9H), 0.02 (s, 6H).

[0312] Step 2: Preparation of compound PE-21E-c

[0313] 5-[(tert-butyldimethylsilyl)oxy]-pentyl-4-nitrophenyl carbonate (5.57 mg, 14.53 μmol, 2 eq) was dissolved in THF (500 μL) and cooled to 0 °C, then eribulin mesylate (6 mg, 7.26 μmol, 1 eq) and DIPEA (2.82 mg, 3.60 μL, 21.79 μmol, 3 eq) were added. The reaction was reacted at 25 °C for 36 h. LCMS showed that the reaction was complete, and the reaction was concentrated to give the crude product PE-21E-c (8 mg) as a yellow oil.

[0314] LCMS (ESI): m / z, 974.6 [M+H] + Rt = 3.83 min.

[0315] Step 3: Preparation of compound PE-21E

[0316] PE-21E-c (8 mg, 8.21 pmol, 1 eq) was dissolved in DCM (500 pL) and cooled to 0 °C, then TFA (153.50 mg, 100 pL) was added. The reaction was stirred at 0 °C for 30 min. LCMS showed the reaction was complete. The reaction was concentrated and the residue was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30-60% elution) to give PE-21E (1.03 mg, yield 14.59%, purity 97.91%) as a white solid.

[0317] LCMS (ESI): m / z, 882.6 [M+Na] + Rt = 2.208 min.

[0318] Preparation Example 18: Preparation of compound PE-21EA

[0319] Step 1: Preparation of compound PE-21EA-a

[0320] To a solution of compound (9H-fluoren-9-yl)methyl (5-hydroxypentyl)carbamate (895 mg, 2.75 mmol, 1 eq) in DMF (10 mL) was added di(p-nitrophenyl) carbonate (2.59 g, 8.25 mmol, 3 eq) and DIPEA (2.15 g, 16.51 mmol, 6 eq), the reaction was stirred at 25 °C for 1 h. LCMS showed product formation. The reaction was concentrated and purified by silica gel column chromatography (eluent: petroleum ether: ethyl acetate = 0: 100-40:60) and silica gel column chromatography (C18, ISCO, R-220g SepaFlash Silica Flash Column, elution solvent was water / CH3CN, acetonitrile elution ratio was 5-80%, flow rate 80 mL / min) to give PE-21EA-a (1.07 g, yield 79.29%) as a white solid.

[0321] LCMS (ESI): m / z, 491.3 [M+H] + .

[0322] Step 2: Preparation of compound PE-21EA-b

[0323] To a solution of compound eribulin mesylate (45 mg, 54.48 μmol, 1 eq) and DIPEA (21.34 mg, 163.44 μmol, 3 eq) in DMF (1 mL) was added a solution of compound PE-21EA-a (32.07 mg, 65.37 μmol, 1.2 eq) in DMF (1 mL) at 0 °C. The reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was complete. The reaction was filtered and purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860, column Synergi Max-RP 200 x 30 mm x 10 μm, mobile phase water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution) and silica gel column chromatography (eluent: dichloromethane: tetrahydrofuran = 0: 100 ~ 30: 70) to give PE-21EA-b (56.10 mg, yield 95.23%) as a white solid.

[0324] LCMS (ESI): m / z, 1103.8 [M+Na] + .

[0325] Step 3: Preparation of compound PE-21EA

[0326] To a solution of compound PE-21EA-b (42 mg, 38.84 μmol, 1 eq) in DMF (3 mL) was added diethylamine (29 mg, 388.42 μmol, 10 eq) at 0 °C. The reaction was stirred at 25 °C for 1 h. LCMS showed the starting material was consumed completely. The reaction was filtered and purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860, column Synergi Max-RP 200 x 30 mm x 10 μm, mobile phase water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution) to give PE-21EA (26.06 mg, yield 78.1%, purity 95%) as a white solid.

[0327] LCMS (ESI): m / z, 859.6 [M+H] + .

[0328] 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 6.94 (t, J = 5.9 Hz, 1H), 5.05 (s, 1H), 4.99 (s, 1H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (d, J = 3.9 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.9 Hz, 1H), 4.18 (dd, J = 10.3, 4.2 Hz, 1H), 4.09 (d, J = 5.0 Hz, 2H), 4.02 (t, J = 9.6 Hz, 1H), 3.91 (t, J = 6.6 Hz, 2H), 3.80 (s, 1H), 3.77 - 3.73 (m, 1H), 3.70 (s, 1H), 3.56 (s, 2H), 3.49 (d, J = 10.7 Hz, 5H), 3.25 (s, 4H), 2.94 (t, J = 6.0 Hz, 2H), 2.84 (d, J = 9.6 Hz, 1H), 2.79 - 2.73 (m, 1H), 2.69 (q, J = 6.5, 5.6 Hz, 3H), 2.60 - 2.54 (m, 1H), 2.34 - 2.30 (m, 1H), 2.24 (t, J = 14.7 Hz, 3H), 2.14 (d, J = 12.0 Hz, 1H), 2.04 - 1.95 (m, 2H), 1.91 (d, J = 9.0 Hz, 4H), 1.71 (d, J = 11.4 Hz, 2H), 1.65 (d, J = 12.6 Hz, 2H), 1.50 (dd, J = 14.7, 7.5 Hz, 5H), 1.35 - 1.30 (m, 3H), 1.26 (d, J = 12.9 Hz, 2H), 1.19 - 1.12 (m, 1H), 1.03 (d, J = 6.6 Hz, 3H), 0.94 (d, J = 6.8 Hz, 1H), 0.86 (d, J = 6.9 Hz, 1H), 0.84 - 0.80 (m, 2H).

[0329] Preparation Example 19: Preparation of compound PE-21 EMS

[0330] Step 1: Preparation of compound (S)-tert-butyldimethylsilyl-2-methylbutanol

[0331] To a solution of compound (4S)-1,4-pentanediol (100 mg, 960.15 pmol, 1 eq) in DCM (2 mL) was added compound imidazole (65.37 mg, 960.15 pmol, 1 eq) and tert-butyldimethylsilyl chloride (159.19 mg, 1.06 mmol, 1.1 eq) at 0 °C. The reaction was stirred at room temperature overnight. TLC showed the reaction was completed. To the mixture was added DCM (10 mL) and water (20 mL), the mixture was extracted with DCM (20 mL) for 3 times, the combined organic phase was washed with saturated brine (10 mL), dried over anhydrous Na2S04, filtered and concentrated to give the product (S)-tert-butyldimethylsilyoxy-2-methylbutanol (131 mg, yield 62.47%) as colorless oil.

[0332] 1 H NMR (400 MHz, CDC13) δ 8.29 (d, J = 9.2 Hz, 2H), 7.42 - 7.36 (m, 2H), 5.03 - 4.89 (m, 1 H), 3.74 - 3.60 (m, 2H), 1.85 - 1.58 (m, 4H), 1.45 - 1.33 (m, 3H), 0.95 - 0.86 (m, 9H), 0.10 - 0.02 (m, 6H).

[0333] Step 2: Preparation of compound PE-21 EMS-a

[0334] To a solution of compound (S)-tert-butyldimethylsilyoxy-2-methylbutanol (131 mg, 599.79 pmol, 1 eq) in DMF (2 mL) was added compound NPC (547.39 mg, 1.80 mmol, 3 eq) and DIPEA (465.10 mg, 594.76 pL, 3.60 mmol, 6 eq), the reaction was stirred at 35 °C for 2 hours. LCMS showed the reaction was completed. The reaction was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether 0:1 to 1 :20) to give the product PE-21 EMS-a (112 mg, yield 48.69%) as colorless oil.

[0335] 1 H NMR (400 MHz, CDC13) δ 8.29 (d, J = 9.2 Hz, 2H), 7.42 - 7.36 (m, 2H), 5.03 - 4.89 (m, 1 H), 3.74 - 3.60 (m, 2H), 1.85 - 1.58 (m, 4H), 1.45 - 1.33 (m, 3H), 0.95 - 0.86 (m, 9H), 0.10 - 0.02 (m, 6H).

[0336] Step 3: Preparation of compound PE-21 EMS-b

[0337] To a solution of compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq) and PE-21 EMS-a (5.57 mg, 14.53 pmol, 1.2 eq) in DMF (2 mL), compound DIPEA (4.69 mg, 36.32 pmol, 3 eq) and HOBt (1.64 mg, 12.11 pmol, 1 eq) were added, and the reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The reaction was rotary evaporated under reduced pressure to give the product PE-21 EMS-b (11.80 mg, crude) as colorless oil. LCMS (ESI): m / z, 1014.6 [M+Na] + Rt = 2.968 min.

[0338] Step 4: Preparation of compound PE-21 EMS

[0339] To a solution of compound PE-21 EMS-b (11.4 mg, crude) in DMF (2 mL), triethylamine trifluoride (0.1 mL) was added, and the reaction was stirred at 65 °C for 2.5 hours. LCMS showed the starting material was completely reacted. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860-R6. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) to give PE-21 EMS (5.10 mg, yield 50.68%) as a white solid. LCMS (ESI): m / z, 860.6 [M+H] + Rt = 2.038 min.

[0340] 1 H NMR (400 MHz, DMSO-d6) d 6.96 - 6.80 (m, 1H), 5.02 (d, J = 22.8 Hz, 2H), 4.79 (d, J = 31.2 Hz, 2H), 4.67 - 4.50 (m, 3H), 4.44 - 4.36 (m, 1H), 4.29 - 3.64 (m, 10H), 3.61 - 3.39 (m, 5H), 3.25 (s, 6H), 2.99 - 2.53 (m, 7H), 2.36 - 1.88 (m, 11H), 1.78 - 1.10 (m, 17H), 1.06 - 0.92 (m, 4H).

[0341] Preparation Example 20: Preparation of compound PE-21 EMR

[0342] Step 1: Preparation of compound (R)-tert-butyldimethylsilyl-2-methylbutanol

[0343] To a solution of compound (4R)-1,4-pentanediol (50 mg, 480.08 pmol, 1 eq) in DCM (2 mL) was added compound imidazole (32.68 mg, 480.08 pmol, 1 eq) and tert-butyldimethylsilyl chloride (79.59 mg, 528.08 pmol, 1.1 eq) at 0 °C. The reaction was stirred at room temperature overnight. TLC showed the reaction was completed. To the mixture was added DCM (10 mL) and water (20 mL), the mixture was extracted with DCM (20 mL for each) for 3 times, the combined organic phase was washed with saturated brine (10 mL), dried over anhydrous Na2S04, filtered and concentrated to give the product (R)-tert-butyldimethylsilyoxy-2-methylbutanol (84.40 mg, yield 80.49%) as colorless oil.

[0344] 1 H NMR (400 MHz, CDC13) 3.93-3.56 (m, 3H), 1.70-1.60 (m, 2H), 1.53-1.42 (m, 2H), 1.27-1.1 1 (m, 3H), 0.94-0.82 (m, 10H), 0.1 1 (s, 3H), 0.08 (s, 3H).

[0345] Step 2: Preparation of compound PE-21 EMR-a

[0346] To a solution of compound (R)-tert-butyldimethylsilyoxy-2-methylbutanol (84.40 mg, 386.43 pmol, 1 eq) in DMF (2 mL) was added compound NPC (352.67 mg, 1.16 mmol, 3 eq) and DIPEA (299.65 mg, 2.32 mmol, 6 eq), the reaction was stirred at 35 °C for 2 hours. LCMS showed the reaction was completed. The reaction was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether 0:1 to 1 :20) to give the product PE-21 EMR-a (40 mg, yield 26.99%) as colorless oil.

[0347] 1 H NMR (400 MHz, CDC13) 3.93-3.56 (m, 3H), 1.70-1.60 (m, 2H), 1.53-1.42 (m, 2H), 1.27-1.1 1 (m, 3H), 0.94-0.82 (m, 10H), 0.1 1 (s, 3H), 0.08 (s, 3H).

[0348] Step 3: Preparation of compound PE-21 EMR-b

[0349] To a solution of compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq) and PE-21 EMR-a (6 mg, 15.64 pmol, 1.29 eq) in DMF (2 mL), compound DIPEA (4.69 mg, 36.32 pmol, 3 eq) and HOBt (1.64 mg, 12.11 pmol, 1 eq) were added, and the reaction was stirred at room temperature for 1 h. LCMS showed the reaction was completed. The reaction was rotary evaporated under reduced pressure to give the product PE-21 EMR-b (11.80 mg, crude) as a colorless oil. LCMS (ESI): m / z, 1014.6 [M+Na] + .

[0350] Step 4: Preparation of compound PE-21 EMR

[0351] To a solution of compound PE-21 EMR-b (11.8 mg, crude) in DMF (2 mL), triethylamine trifluoride (0.1 mL) was added, and the reaction was stirred at 60 °C for 4 h. LCMS showed the starting material was completely consumed. The mixture was purified by high performance liquid chromatography-preparative (Oriendo, model BRIX-2860-R6. Column: Welch Xtimate C18 250 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give PE-21 EMR (3.90 mg, yield 37.44%) as a white solid. LCMS (ESI): m / z, 860.6 [M+H] + .

[0352] 1 H NMR (400 MHz, DMSO-d6) d 6.93 - 6.81 (m, 1H), 5.02 (d, J = 22.8 Hz, 2H), 4.79 (d, J = 31.6 Hz, 2H), 4.68 - 4.49 (m, 3H), 4.43 - 4.35 (m, 1H), 4.31 - 3.63 (m, 9H), 3.62 - 3.37 (m, 6H), 3.30 - 3.23 (m, 6H), 2.96 - 2.55 (m, 7H), 2.35 - 1.89 (m, 11H), 1.77 - 1.08 (m, 17H), 1.07 - 0.92 (m, 4H).

[0353] Preparation Example 21: Preparation of compound PE-21 EK

[0354] Step 1: Preparation of compound PE-21 EK-a

[0355] Compound 5-[(tert-butyldimethylsilyl)oxy]pentyl 4-nitrophenyl carbonate (10 mg, 26.07 μmol, 1 eq, the synthesis of this compound please refer to the preparation example of PE-21E) was dissolved in DMF (2 mL), eribulin mesylate (80 mg, 289.38 μmol, 1 eq) and DIPEA (10.11 mg, 99%, 12.93 μL, 78.22 μmol, 3 eq), HOBT (3.52 mg, 99.96%, 2.35 μL, 26.07 μmol, 1 eq) were added, the reaction was stirred at 25 °C for 2 hours. LCMS showed that the product was generated. The reaction was prepared by high performance liquid chromatography to obtain white solid product (15 mg, yield 59.04%).

[0356] The high performance liquid chromatography preparation conditions were as follows: preparation chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 250x30mmx10μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 65% to 95%.

[0357] LCMS (ESI): m / z, 974.7 [M+H] + .

[0358] Step 2: Preparation of compound PE-21EK-b

[0359] Compound PE-22EK-a (10 mg, 10.26 μmol, 1 eq) was dissolved in DCM (3 mL) solution, DMP (26.12 mg, 98%, 19.18 μL, 61.58 μmol, 6 eq) was added at 0 °C, and finally a catalytic amount of water (50 μL) was added, and stirred at 25 °C for 12 hours. LCMS showed that the reaction was complete. The reaction was rotary dried, the residue was dissolved in DMF (1 mL), and prepared by high performance liquid chromatography to obtain white solid product (3 mg, yield 30.06%).

[0360] The high performance liquid chromatography preparation conditions were as follows:

[0361] Preparation chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150x30mmx10μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of water was from 65% to 95%.

[0362] LCMS (ESI): m / z, 972.54 [M+H] + .

[0363] Step 3: Preparation of compound PE-21EK

[0364] To the solution of compound PE-21EK-b (3 mg, 3.09 pmol, 1 eq) in THF (1 mL) was added Et3N.3HF (98.90 mg, 97%, 100 pL, 595.08 pmol, 192.8653 eq) at 25 °C. The reaction was stirred at 25 °C for 2 h. LCMS showed the starting material was consumed completely. The reaction was purified by high performance liquid chromatography to give the product (1.32 mg, yield 49.86%, purity 96%) as a white solid.

[0365] The high performance liquid chromatography preparation conditions were as follows:

[0366] The preparation chromatograph factory was Oriendo, model BRIX-2860. The chromatograph column was YMC-Triart Prep C18 150*30 mm*10 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the acetonitrile elution ratio was eluted from 30% to 60%.

[0367] LCMS (ESI): m / z, 880.4 [M+Na] + .

[0368] 1 H NMR (400 MHz, DMSO-d6) d 7.29 (t, J = 6.0 Hz, 1H), 5.04 (d, J = 11.1 Hz, 2H), 4.83 (s, 1H), 4.77-4.73 (m, 1H), 4.63 (d, J = 2.8 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.37 (t, J = 5.1 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (td, J = 10.2, 4.2 Hz, 1H), 4.10 (t, J = 3.2 Hz, 2H), 4.06-3.98 (m, 2H), 3.93 (t, J = 6.7 Hz, 2H), 3.87-3.75 (m, 3H), 3.71 (s, 1H), 3.57-3.47 (m, 2H), 3.23 (s, 3H), 2.84 (d, J = 9.2 Hz, 1H), 2.82-2.72 (m, 2H), 2.71-2.65 (m, 2H), 2.60 (dd, J = 18.0, 8.0 Hz, 2H), 2.29 (q, J = 18.7, 17.8 Hz, 5H), 2.18-1.84 (m, 8H), 1.68 (q, J = 12.5 Hz, 4H), 1.58-1.38 (m, 6H), 1.38-1.15 (m, 6H), 1.00 (dd, J = 22.6, 9.1 Hz, 4H).

[0369] Preparation Example 22: Preparation of compound E2KO

[0370] Step 1: Preparation of compound PE-E2KO

[0371] Compound PE-E2K (30 mg, 0.04 mmol) was dissolved in N,N-dimethylformamide (2 mL), hydroxyacetic acid (7 mg, 0.08 mmol) and 1-hydroxybenzotriazole (16 mg, 0.12 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (23 mg, 0.12 mmol) were added at room temperature, the reaction solution was stirred at 30 °C for 2 hours, the reaction solution was prepared by high performance liquid chromatography (Prep HPLC, Oriendo, BRIX-2860. The column was Phenomenex Luna C18 250 x 50 mm x 10 μm. The mobile phase was water (0.5% TFA) - acetonitrile, the elution ratio of acetonitrile was eluted from 12% to 42%), the eluate was lyophilized to obtain white solid product PE-E2KO (8.1 mg, 24.7%).

[0372] LCMS (ESI): m / z, 792.5 [M+H] + Rt = 0.97 min.

[0373] 1 H NMR (400 MHz, DMSO-d 6) δ 7.51 (s, 1H), 4.64 (t, J = 4.4 Hz, 1H), 4.56 (s, 1H), 4.39 (s, 1H), 4.11 (d, J = 1.6 Hz, 4H), 3.79 (s, 6H), 3.55 (d, J = 5.2 Hz, 3H), 3.27 (d, J = 3.2 Hz, 2H), 3.25 (s, 3H), 3.21 - 3.01 (m, 3H), 2.89 - 2.61 (m, 4H), 2.43 - 2.12 (m, 6H), 1.95 (d, J = 17.2 Hz, 5H), 1.71 - 1.43 (m, 9H), 1.23 (s, 4H), 0.94 (d, J = 6.4 Hz, 3H).

[0374] Preparation Example 23: Preparation of compound PE-21F

[0375] Step 1: Preparation of 6-((tert-butyldimethylsilyl)oxy)hexyl-4-nitrophenyl carbonate

[0376] 6-((tert-butyldimethylsilyl)oxy)hexan-1-ol (232.43 mg, 1 mmol, 1 eq) was dissolved in DMF (3 mL), then added di(p-nitrophenyl) carbonate (912.63 mg, 608.42 μL, 3 mmol, 3 eq) and DIPEA (387.72 mg, 495.81 μL, 3 mmol, 3 eq). The reaction was reacted at 35 °C for 2 hours. LCMS showed that a new compound was generated. The reaction was diluted with water (50 mL) and extracted with dichloromethane (50 mL x 3). The organic phase was combined, washed with brine (100 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (eluent ratio: PE / EA = 95% / 5%) to obtain 6-((tert-butyldimethylsilyl)oxy)hexyl-4-nitrophenyl carbonate (370 mg, yield 93.07%) as a yellow oil.

[0377] 1 H NMR (400 MHz, DMSO-d6) δ 8.37-8.24 (m, 2H), 7.54 (dd, J = 9.2, 1.6 Hz, 2H), 4.23 (t, J = 6.4 Hz, 2H), 3.56 (t, J = 6.4 Hz, 2H), 1.67 (t, J = 7.2 Hz, 2H), 1.46 (t, J = 6.4 Hz, 2H), 1.34 (dt, J = 8.0, 4.8 Hz, 4H), 0.84 (d, J = 1.2 Hz, 9H), 0.01 (d, J = 1.2 Hz, 6H).

[0378] Step 2: Preparation of compound PE-21F-c

[0379] 6-((tert-butyldimethylsilyl)oxy)hexyl-4-nitrophenyl carbonate (9.63 mg, 24.21 μmol, 2 eq) was dissolved in THF (500 μL) and cooled to 0 °C, then added eribulin mesylate (10 mg, 12.11 μmol, 1 eq) and DIPEA (4.69 mg, 6 μL, 36.32 μmol, 3 eq). The reaction was reacted at 25 °C for 16 hours. LCMS showed that the reaction was complete. The reaction was concentrated to obtain the crude yellow oil PE-21F-c (12 mg).

[0380] LCMS (ESI): m / z, 988.7 [M+H] + Rt = 3.93 min.

[0381] Step 3: Preparation of compound PE-21F

[0382] Compound PE-21F-c (12 mg, 12.14 μmol, 1 eq) was dissolved in DCM (500 μL), cooled to 0 °C, then added TFA (153.50 mg, 100 μL). The reaction was reacted at 0 °C for 30 min. LCMS showed the reaction was complete, the reaction was concentrated, the residue was prepared by high performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was Welch Xtimate C18 250 x 40 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution), to obtain white solid PE-21F (2.46 mg, yield 23.18%, purity 100%).

[0383] LCMS (ESI): m / z, 874.6 [M+H] + Rt = 2.298 min.

[0384] Preparation Example 24: Preparation of compound PE-21G

[0385] Step 1: Preparation of compound 3-((tert-butyldimethylsilyl)oxy)-1- cyclopropylpropan-1-ol

[0386] To a solution of compound 1-cyclopropylpropane-1,3-diol (95 mg, 817.84 μmol, 1 eq) in DCM (8 mL) was added TBSCl (135.59 mg, 155.85 μL, 899.62 μmol, 1.1 eq) and imidazole (55.68 mg, 50.62 μL, 817.84 μmol, 1 eq) at 0 °C, after addition, 25 °C was reacted for 16 h, nuclear magnetic resonance showed that the reaction was complete, the reaction was poured into ice sodium bicarbonate aqueous solution (60 mL), then stirred for 2 min, extracted with dichloromethane (60 mL x 3), the organic phase was combined and washed with brine, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated to obtain anhydrous oil compound 3-((tert-butyldimethylsilyl)oxy)-1-cyclopropylpropan-1-ol (147 mg, yield 78.01%).

[0387] 1H NMR (400 MHz, Chloroform-d) δ 3.96-3.87 (m, 1H), 3.80 (ddd, J = 10.1, 7.0, 6.0 Hz, 1H), 3.11 (dt, J = 8.3, 5.8 Hz, 1H), 1.85-1.78 (m, 2H), 0.89 (s, 9H), 0.89-0.86 (m, 1H), 0.57-0.41 (m, 2H), 0.41-0.33 (m, 1H), 0.21-0.13 (m, 1H), 0.07 (d, J = 2.9 Hz, 6H).

[0388] Step 2: Preparation of compound PE-21G-a

[0389] To a solution of 3-((tert-butyldimethylsilyl)oxy)-1-cyclopropylpropan-1-ol (304 mg, 1.32 mmol, 1 eq) in DMF (10 mL) was added di(p-nitrophenyl) carbonate (1.61 g, 1.07 mL, 5.28 mmol, 4 eq) and DIPEA (1.02 g, 1.31 mL, 7.92 mmol, 6 eq) at 0 °C. The reaction was stirred at 33 °C for 2 h. LCMS showed the formation of a new compound. The reaction was concentrated to dryness to give a residue, which was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:30 to 1:20) to give PE-21G-a (257 mg, yield 49.25%) as a light yellow oil.

[0390] 1 H NMR (400 MHz, Chloroform-d) δ 8.29-8.26 (m, 2H), 7.41-7.36 (m, 2H), 4.38 (ddd, J = 9.3, 7.8, 5.1 Hz, 1H), 3.81-3.69 (m, 2H), 2.11-1.92 (m, 2H), 1.18-1.06 (m, 1H), 0.88 (s, 9H), 0.67-0.55 (m, 3H), 0.40-0.31 (m, 1H), 0.05 (d, J = 1.1 Hz, 6H). The structure of this compound was further confirmed by 2D spectra.

[0391] Step 3: Preparation of compound PE-21G-b

[0392] To a solution of compound PE-21G-a (5.57 mg, 14.53 pmol, 2 eq) in DMF (1 mL) was added eribulin mesylate (10 mg, 12.11 pmol, 1 eq) and DIPEA (4.69 mg, 6 pL, 36.32 pmol, 3 eq) and HOBt (1.64 mg, 1.09 pL, 12.11 pmol, 1 eq) at 0 °C. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was complete. The reaction was concentrated to give the crude product PE-21G-b (12 mg, crude) as yellow oil.

[0393] LCMS (ESI): m / z, 1008.6 [M+Na] + Rt = 2.430 min.

[0394] Step 4: Preparation of compound PE-21G

[0395] To a solution of compound PE-21G-b (12 mg, 12.17 pmol, 1 eq) in THF (3 mL) was added triethylamine trihydrofluoride (78.45 mg, 79.33 pL, 486.65 pmol, 40 eq) at room temperature. The reaction was stirred at 60 °C for 2 h. LCMS showed the reaction was complete. The reaction was concentrated. The residue was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30-60% elution) to give PE-21G (3.77 mg, yield 35.53%, purity 95%) as white solid after lyophilization.

[0396] LCMS (ESI): m / z, 894.5 [M+Na] + Rt = 2.168 min.

[0397] 1H NMR (400 MHz, DMSO-d6) δ 6.86 (t, J = 5.8 Hz, 1H), 5.05 (s, 1H), 4.99 (s, 1H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (dt, J = 4.3, 2.4 Hz, 1H), 4.58 - 4.47 (m, 2H), 4.39 (td, J = 5.1, 3.3 Hz, 1H), 4.26 (d, J = 10.9 Hz, 1H), 4.22 - 4.05 (m, 5H), 4.02 (s, 1H), 3.84 - 3.65 (m, 3H), 3.59 - 3.42 (m, 5H), 3.26 (s, 4H), 3.01 - 2.89 (m, 2H), 2.84 (d, J = 9.8 Hz, 1H), 2.72 (td, J = 20.0, 18.2, 8.7 Hz, 2H), 2.61 - 2.53 (m, 1H), 2.23 (tt, J = 29.1, 12.9 Hz, 6H), 1.96 (d, J = 34.1 Hz, 6H), 1.81 - 1.58 (m, 7H), 1.56 - 1.41 (m, 3H), 1.38 - 1.12 (m, 4H), 1.07 - 0.83 (m, 5H), 0.51 - 0.34 (m, 2H), 0.34 - 0.19 (m, 2H).

[0398] Preparation Example 25: Preparation of compound PE-AL1

[0399] Step 1: Preparation of compound PE-AL1-a

[0400] To a solution of compound PE-AM1-b (synthesis of this compound please refer to the preparation example of PE-AM1) (16 mg, 22.89 μmol, 1 eq) and 3-N-tert-butoxycarbonyl aminocyclobutane (4.88 mg, 97%, 27.47 μmol, 1.2 eq) in 1,2-dichloroethane (1 mL) was added acetic acid (2.76 mg, 99.5%, 2.64 μL, 45.79 μmol, 2 eq), the reaction was stirred at 25 °C for 20 min, then sodium triacetoxyborohydride (9.90 mg, 98%, 45.79 μmol, 2 eq) was added, and stirred at 25 °C for 40 min, LCMS showed that the reaction was completed, the reaction was added with water (20 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (30 mL), dried with anhydrous sodium sulfate, and rotary evaporated under reduced pressure to give white solid compound PE-AL1-a (26 mg, crude).

[0401] LCMS (ESI): m / z, 855.6 [M+H] + .

[0402] Step 2: Preparation of compound PE-AL1

[0403] To a solution of compound PE-AL1-a (19.58 mg, 22.90 pmol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.2 mL) at 0 °C and stirred at 0 °C for 1 h. LCMS showed the reaction was completed. The reaction solution was concentrated and purified by high performance liquid chromatography-preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give compound PE-AL1 (0.93 mg, yield 5.38%) as a white solid after lyophilization.

[0404] LCMS (ESI): m / z, 755.5 [M+H] + , 777.5 [M+Na] + .

[0405] 1H NMR (400 MHz, DMSO-d6) δ 6.20 (d, J = 3.0 Hz, 1H), 5.95 (d, J = 3.0 Hz, 1H), 4.96 - 4.94 (m, 1H), 4.87 (s, 1H), 4.81 (s, 1H), 4.75 (s, 1H), 4.34 (d, J = 6.1 Hz, 1H), 4.30 (d, J = 9.6 Hz, 1H), 4.10 - 4.04 (m, 1H), 4.01 (d, J = 2.6 Hz, 1H), 3.78 (dd, J = 9.7, 2.7 Hz, 1H), 3.68 (q, J = 4.7, 3.7 Hz, 3H), 3.59 (dd, J = 9.0, 4.6 Hz, 3H), 3.54 - 3.49 (m, 6H), 3.22 (s, 4H), 2.97 (d, J = 7.1 Hz, 1H), 2.80 - 2.74 (m, 2H), 2.67 - 2.62 (m, 3H), 2.58 - 2.55 (m, 1H), 2.43 (d, J = 9.4 Hz, 3H), 2.32 (d, J = 4.6 Hz, 1H), 2.25 (t, J = 6.5 Hz, 1H), 2.14 - 2.08 (m, 1H), 2.01 - 1.94 (m, 3H), 1.85 (dd, J = 11.9, 4.3 Hz, 1H), 1.70 (dd, J = 11.0, 6.4 Hz, 3H), 1.62 (dd, J = 10.4, 5.0 Hz, 2H), 1.53 (d, J = 7.1 Hz, 2H), 1.48 - 1.40 (m, 3H), 1.34 - 1.28 (m, 3H), 1.04 (d, J = 6.5 Hz, 3H), 0.97 - 0.91 (m, 1H), 0.85 (t, J = 6.7 Hz, 1H).

[0406] Preparation Example 26: Preparation of compound PE-AL2

[0407] Step 1: Preparation of compound PE-AL2-a

[0408] To a solution of compound PE-AM1-b (synthesis of this compound please refer to the preparation of PE-AM1) (16 mg, 22.89 umol, 1 eq) and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (4.78 mg, 95%, 22.89 umol, 1 eq) in 1,2-dichloroethane (1 mL) was added acetic acid (2.76 mg, 99.5%, 2.64 uL, 45.79 umol, 2 eq), the reaction was stirred at 25 °C for 20 min, then sodium triacetoxyborohydride (9.90 mg, 98%, 45.79 umol, 2 eq) was added, and stirred at 25 °C for 40 min, LCMS showed the reaction was completed, the reaction was added water (20 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and rotary evaporated under reduced pressure to give white solid compound PE-AL2-a (24 mg, crude).

[0409] LCMS (ESI): m / z, 881.6 [M+H] + , 903.6 [M+Na] + .

[0410] Step 2: Preparation of compound PE-AL2

[0411] To a solution of compound PE-AL2-a (20.17 mg, 22.89 umol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.2 mL) at 0 °C, and stirred at 25 °C for 1 h, LCMS showed the reaction was completed, the reaction was concentrated, then purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 um. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile was eluted from 30% to 60% elution), and lyophilized to give white solid compound PE-AL2 (2.32 mg, yield 12.98%).

[0412] LCMS (ESI): m / z, 781.4 [M+H] + , 803.4 [M+Na] + .

[0413] 1H NMR (400 MHz, DMSO-d6) δ 6.20 (d, J = 3.0 Hz, 1H), 5.95 (d, J = 3.0 Hz, 1H), 4.95 (d, J = 2.1 Hz, 1H), 4.88 - 4.85 (m, 1H), 4.80 (s, 1H), 4.75 (s, 1H), 4.34 (d, J = 6.3 Hz, 1H), 4.29 (d, J = 9.7 Hz, 1H), 4.06 (t, J = 6.6 Hz, 1H), 4.00 (t, J = 2.5 Hz, 1H), 3.87 (d, J = 9.7 Hz, 4H), 3.78 (dd, J = 9.7, 2.7 Hz, 1H), 3.68 (dd, J = 9.8, 5.9 Hz, 4H), 3.57 (dt, J = 9.3, 5.0 Hz, 6H), 3.21 (s, 5H), 3.00 - 2.95 (m, 2H), 2.80 - 2.73 (m, 2H), 2.66 (dt, J = 6.8, 1.9 Hz, 1H), 2.63 - 2.57 (m, 2H), 2.56 - 2.54 (m, 1H), 2.44 (d, J = 3.4 Hz, 1H), 2.33 (ddd, J = 7.8, 5.6, 3.7 Hz, 3H), 2.24 (t, J = 7.6 Hz, 1H), 2.14 - 2.08 (m, 1H), 1.97 - 1.90 (m, 2H), 1.85 (dd, J = 12.0, 3.9 Hz, 1H), 1.71 (ddd, J = 14.0, 10.3, 6.5 Hz, 4H), 1.65 - 1.57 (m, 3H), 1.52 - 1.47 (m, 2H), 1.45 - 1.39 (m, 2H), 1.34 - 1.27 (m, 2H), 1.04 (d, J = 6.5 Hz, 3H), 0.98 - 0.90 (m, 1H).

[0414] Preparation Example 27: Preparation of compound PE-AL2H5

[0415] To a solution of compound PE-AM1-b (synthesis of this compound please refer to the preparation of PE-AM1) (16 mg, 22.89 μmol, 1 eq) and ammonia water (3.85 mg, 25%, 27.47 μmol, 1.2 eq) in 1,2-dichloroethane (1 mL) was added acetic acid (2.76 mg, 99.5%, 2.64 μL, 45.79 μmol, 2 eq). After the reaction solution was stirred at 25 °C for 20 min, sodium triacetoxyborohydride (9.90 mg, 98%, 45.79 μmol, 2 eq) was added and stirred at 25 °C for 40 min. LCMS showed the reaction was completed. The reaction solution was concentrated and purified by high performance liquid chromatography-preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give compound PE-AL2H5 (0.42 mg, 2.62% yield) as a white solid after lyophilization.

[0416] LCMS (ESI): m / z, 700.2 [M+H] + , 722.2 [M+Na] + .

[0417] 1H NMR (400 MHz, DMSO-d6) δ 5.06 (s, 1H), 4.98 (s, 1H), 4.83 (s, 1H), 4.76 (s, 1H), 4.63 (dt, J = 4.6, 2.4 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.18 (dd, J = 10.5, 4.0 Hz, 1H), 4.10 (d, J = 4.8 Hz, 3H), 4.02 (s, 1H), 3.80 (t, J = 10.1 Hz, 1H), 3.74 - 3.67 (m, 2H), 3.48 (s, 5H), 3.27 (s, 2H), 3.26 (s, 1H), 2.84 (dd, J = 9.5, 1.9 Hz, 1H), 2.74 (q, J = 7.8, 6.9 Hz, 2H), 2.67 (h, J = 2.3, 1.9 Hz, 1H), 2.60 - 2.54 (m, 1H), 2.36 - 2.06 (m, 6H), 2.04 - 1.97 (m, 1H), 1.96 - 1.87 (m, 3H), 1.86 - 1.76 (m, 1H), 1.76 - 1.61 (m, 4H), 1.56 - 1.43 (m, 2H), 1.33 (dd, J = 29.9, 18.2 Hz, 3H), 1.23 (s, 2H), 1.22 - 1.05 (m, 2H), 1.03 (d, J = 6.5 Hz, 2H), 1.00 - 0.93 (m, 1H), 0.89 - 0.80 (m, 2H).

[0418] Preparation Example 28: Preparation of compound PE-AL3

[0419] Step 1: Preparation of compound PE-AL3-a

[0420] To a solution of compound PE-AM1-b (16 mg, crude, its synthesis refer to the preparation of PE-AM1) and tert-butyl N-2-azaspiro[3.3]heptan-6-ylcarbamate (5.01 mg, 22.89 pmol, 1 eq) in DCE (1 mL) was added acetic acid (2.76 mg, 45.79 pmol, 2 eq) and the reaction was stirred at room temperature for 20 minutes. To the reaction was added sodium triacetoxyborohydride (9.90 mg, 45.79 pmol, 2 eq) and the reaction was stirred at room temperature for 40 minutes. LCMS showed the reaction was complete. To the mixture was added DCM (10 mL) and water (20 mL) and the mixture was extracted with DCM (20 mL for each) for 3 times. The combined organic phase was washed with saturated brine (10 mL), dried over anhydrous Na2S04, filtered and concentrated to give the product PE-AL3-a (20.59 mg, crude, yield not calculated) as a colorless oil. LCMS (ESI): m / z, 895.6 [M+H] + .

[0421] Step 2: Preparation of compound PE-AL3

[0422] To a solution of compound PE-AL3-a (20.49 mg, crude) in DCM (2 mL) was added TFA (0.4 mL) at 0 °C and the reaction was stirred at room temperature for half an hour. LCMS showed the starting material was consumed completely. The mixture was concentrated under reduced pressure to give the crude product which was purified by high performance liquid chromatography preparation (Oriendo, model BRIX-2860-R6. Column: Welch Xtimate C18 250 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution) to give PE-AL3 (2.70 mg, yield 12.37%) as a white solid. LCMS (ESI): m / z, 795.6 [M+H] + .

[0423] 1H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 2H), 6.20 (d, J = 3.2 Hz, 1H), 5.95 (d, J = 3.2 Hz, 1H), 4.95 (s, 1H), 4.89 - 4.72 (m, 4H), 4.34 (d, J = 6.4 Hz, 1H), 4.29 (d, J = 9.6 Hz, 1H), 4.11 - 3.94 (m, 4H), 3.80 - 3.75 (m, 3H), 3.70 (d, J = 4.0 Hz, 3H), 3.33 (d, J = 3.6 Hz, 3H), 3.21 (s, 2H), 3.18 (s, 1H), 3.16 - 3.11 (m, 3H), 3.04 (d, J = 9.2 Hz, 2H), 2.99 - 2.92 (m, 2H), 2.80 - 2.72 (m, 2H), 2.69 - 2.58 (m, 3H), 2.39 - 2.27 (m, 6H), 2.15 - 2.06 (m, 2H), 1.97 - 1.81 (m, 4H), 1.75 - 1.54 (m, 7H), 1.52 - 1.39 (m, 4H), 1.35 - 1.27 (m, 2H), 1.04 (d, J = 6.4 Hz, 2H), 1.01 - 0.88 (m, 2H).

[0424] Preparation Example 29: Preparation of compound PE-AL4

[0425] To a solution of compound PE-AM1-b (synthesis of this compound please refer to preparation example of PE-AM1) (16 mg, 22.89 μmol, 1 eq) and 3-methoxyazetidine hydrochloride (3.50 mg, 97%, 27.47 μmol, 1.2 eq) in 1,2-dichloroethane (1 mL) was added acetic acid (2.76 mg, 99.5%, 2.64 μL, 45.79 μmol, 2 eq), the reaction was stirred at 25 °C for 20 min, then sodium triacetoxyborohydride (9.90 mg, 98%, 45.79 μmol, 2 eq) was added and stirred at 25 °C for 40 min, LCMS showed the reaction was completed, the reaction was added water (20 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, rotary evaporation under reduced pressure to give the crude product, the crude product was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Synergi Max-RP 200 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile was eluted from 20% to 50% elution), freeze-drying to give white solid compound PE-AL4 (0.59 mg, yield 3.35%).

[0426] LCMS (ESI): m / z, 770.5 [M+H] + .

[0427] 1 H NMR (400 MHz, DMSO-d6) δ 5.03 (dd, J = 18.1, 2.4 Hz, 2H), 4.83 (s, 1H), 4.75 (d, J = 1.7 Hz, 1H), 4.63 (dt, J = 4.4, 2.5 Hz, 1H), 4.55 (t, J = 4.3 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (td, J = 10.3, 4.2 Hz, 1H), 4.10 (t, J = 3.1 Hz, 2H), 4.03 (d, J = 10.3 Hz, 1H), 3.80 (t, J = 10.4 Hz, 1H), 3.67 (td, J = 6.6, 2.7 Hz, 1H), 3.61 (td, J = 6.7, 3.3 Hz, 1H), 3.55 - 3.49 (m, 2H), 3.45 (d, J = 6.7 Hz, 2H), 3.26 (s, 3H), 3.22 (d, J = 3.5 Hz, 1H), 3.15 (s, 2H), 2.85 - 2.76 (m, 3H), 2.75 - 2.65 (m, 2H), 2.60 - 2.52 (m, 1H), 2.42 (q, J = 6.8 Hz, 2H), 2.35 - 2.31 (m, 3H), 2.27 - 2.22 (m, 2H), 2.19 - 2.09 (m, 2H), 2.04 - 1.97 (m, 2H), 1.92 (d, J = 2.9 Hz, 3H), 1.70 - 1.62 (m, 3H), 1.54 - 1.44 (m, 4H), 1.35 - 1.27 (m, 3H), 1.24 (d, J = 3.3 Hz, 3H), 1.20 - 1.13 (m, 1H), 1.03 (d, J = 6.5 Hz, 3H), 1.00 - 0.93 (m, 1H), 0.85 (t, J = 6.7 Hz, 1H).

[0428] Preparation Example 30: Preparation of compound PE-AL5

[0429] Acetic acid (2.76 mg, 99.5%, 2.64 μL, 45.79 μmol, 2 eq) was added to a solution of compound PE-AM1-b (16 mg, 22.89 μmol, 1 eq) and 5-methylaminopentanol (3.39 mg, 95%, 27.47 μmol, 1.2 eq) in 1,2-dichloroethane (1 mL). The reaction mixture was stirred at 25 °C for 20 min, and then sodium triacetoxyborohydride (9.90 mg, 98%, 45.79 μmol, 2 eq) was added. The mixture was stirred at 25 °C for 40 min. LC-MS showed that the reaction was complete. The reaction mixture was concentrated and then prepared by high-performance liquid chromatography (HPLC) (Shimadzu LC-20AP column, Synergi Max-RP). 200×30mm×10μm. Purified by mobile phase water (0.225% HCOOH)-acetonitrile (elution ratio of acetonitrile from 20% to 50%), and lyophilized to give white solid compound PE-AL5 (5.02 mg, yield 26.86%).

[0430] LCMS(ESI): m / z, 800.5 [M+H] + 822.5 [M+Na] + .

[0431] 1H NMR (400 MHz, DMSO-d6) δ 5.02 (d, J = 23.5 Hz, 2H), 4.83 (d, J = 6.3 Hz, 1H), 4.75 (s, 1H), 4.66 - 4.61 (m, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.5 Hz, 1H), 4.17 (td, J = 10.3, 4.1 Hz, 1H), 4.10 (q, J = 4.2 Hz, 3H), 4.03 (s, 1H), 3.80 (t, J = 10.8 Hz, 1H), 3.67 (t, J = 7.5 Hz, 1H), 3.64 - 3.59 (m, 1H), 3.55 - 3.47 (m, 3H), 3.27 (s, 4H), 3.24 - 3.22 (m, 2H), 3.19 (s, 1H), 2.84 (d, J = 9.6 Hz, 1H), 2.78 - 2.67 (m, 2H), 2.60 - 2.54 (m, 1H), 2.38 (d, J = 6.4 Hz, 1H), 2.34 - 2.22 (m, 7H), 2.17 (d, J = 11.9 Hz, 1H), 2.11 (s, 3H), 2.05 - 1.95 (m, 2H), 1.92 (d, J = 3.0 Hz, 3H), 1.84 (d, J = 14.3 Hz, 1H), 1.67 (dt, J = 15.4, 6.3 Hz, 5H), 1.57 - 1.45 (m, 2H), 1.39 (td, J = 13.2, 11.8, 6.6 Hz, 5H), 1.33 - 1.12 (m, 6H), 1.03 (d, J = 6.4 Hz, 3H), 1.01 - 0.84 (m, 1H).

[0432] Preparation Example 31: Preparation of compound PE-ALH5

[0433] To a solution of compound PE-AM1-b (synthesis of this compound please refer to the preparation of PE-AM1) (32 mg, 45.79 pmol, 1 eq) and 5-amino-1-pentanol (5.97 mg, 95%, 54.95 pmol, 1.2 eq) in 1,2-dichloroethane (2 mL) was added acetic acid (5.53 mg, 99.5%, 5.27 pL, 91.58 pmol, 2 eq), the reaction was stirred at 25 °C for 20 min, then sodium triacetoxyborohydride (19.81 mg, 98%, 91.58 pmol, 2 eq) was added, and stirred at 25 °C for 40 min, LCMS showed the reaction was completed, the reaction was added water (20 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, rotary evaporated under reduced pressure to give the crude product, which was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 20% to 50% elution), and lyophilized to give the compound PE-ALH5 (2.37 mg, yield 6.58%) as a white solid.

[0434] LCMS (ESI): m / z, 786.5 [M+H] + , 808.4 [M+Na] + .

[0435] 1 H NMR (400 MHz, DMSO-d 6) δ 5.02 (d, J = 24.8 Hz, 2H), 4.83 (s, 1H), 4.76 (s, 1H), 4.63 (q, J = 3.2, 2.0 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.6 Hz, 1H), 4.20 - 4.14 (m, 1H), 4.10 (d, J = 5.0 Hz, 3H), 4.02 (s, 1H), 3.78 (d, J = 10.0 Hz, 1H), 3.68 (s, 2H), 3.50 (t, J = 10.0 Hz, 4H), 3.27 (s, 4H), 3.20 (s, 1H), 2.84 (d, J = 9.6 Hz, 1H), 2.78 - 2.55 (m, 7H), 2.34 - 2.12 (m, 6H), 2.04 - 1.86 (m, 6H), 1.68 (dd, J = 27.8, 13.1 Hz, 6H), 1.49 - 1.38 (m, 5H), 1.34 - 1.27 (m, 4H), 1.26 - 1.11 (m, 2H), 1.03 (d, J = 6.4 Hz, 3H), 0.96 (t, J = 11.8 Hz, 1H).

[0436] Preparation Example 32: Preparation of compound PE-ALOH

[0437] To a solution of compound PE-AM1-b (synthesis of this compound please refer to Preparation Example of PE-AM1) (16 mg, 22.89 μmol, 1 eq) in methanol (1 mL) and dichloromethane (0.25 mL) was added sodium borohydride (883.78 μg, 98%, 22.89 μmol, 1 eq) at -78 °C and stirred at -78 to 25 °C for 1 hour, LCMS showed the reaction was completed, the reaction solution was concentrated and purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was Welch Xtimate C18 150 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution), freeze-dried to obtain white solid compound PE-ALOH (11.48 mg, yield 71.54%).

[0438] LCMS (ESI): m / z, 723.4 [M+Na] + .

[0439] 1H NMR (400 MHz, DMSO-d6) δ 5.02 (d, J = 21.9 Hz, 2H), 4.84 (s, 1H), 4.75 (s, 1H), 4.63 (d, J = 3.2 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.41 (t, J = 5.3 Hz, 1H), 4.26 (d, J = 10.9 Hz, 1H), 4.17 (dt, J = 10.1, 5.0 Hz, 1H), 4.10 (t, J = 3.4 Hz, 3H), 4.02 (s, 1H), 3.80 (t, J = 10.5 Hz, 1H), 3.70 (t, J = 5.4 Hz, 2H), 3.55 - 3.49 (m, 1H), 3.48 - 3.36 (m, 3H), 3.26 (s, 2H), 3.23 (d, J = 3.4 Hz, 1H), 2.84 (d, J = 10.1 Hz, 1H), 2.73 (td, J = 19.1, 18.3, 7.5 Hz, 2H), 2.60 - 2.52 (m, 1H), 2.23 (ddd, J = 49.8, 24.5, 13.0 Hz, 6H), 1.96 (d, J = 33.1 Hz, 6H), 1.74 - 1.59 (m, 6H), 1.56 - 1.43 (m, 2H), 1.42 - 1.08 (m, 5H), 1.03 (d, J = 6.4 Hz, 3H), 1.00 - 0.90 (m, 1H).

[0440] Preparation Example 33: Preparation of compound PE-AM1

[0441] Step 1: Preparation of compound PE-AM1-b

[0442] To a solution of compound PE-AM1-a (20 mg, 27.36 μmol, 1 eq, purchased from Haoyuan Pharmatech) in tetrahydrofuran (1 mL) and water (0.5 mL) was added sodium periodate (11.76 mg, 99.5%, 3.04 μL, 54.73 μmol, 2 eq), and stirred at 25 °C for 4 hours, LCMS showed that the raw material was completely reacted, the reaction liquid was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (30 mL), dried with anhydrous sodium sulfate, and rotary evaporated under reduced pressure to obtain white solid compound PE-AM1-b (27 mg, crude). LCMS (ESI): m / z, 721.5 [M+Na] + .

[0443] Step 2: Preparation of compound PE-AM1-c

[0444] To a solution of compound PE-AM1-b (19 mg, 27.19 μmol, 1 eq) and 2-methyl-2-butene (21.29 mg, 90%, 32.10 μL, 271.88 μmol, 10 eq) in tert-butanol (1 mL) was added sodium chlorite (18.44 mg, 80%, 7.38 μL, 163.13 μmol, 6 eq) and sodium phosphate monobasic (6.59 mg, 99%, 54.38 μmol, 2 eq) in water (1 mL) at 0 °C. After addition, the reaction was stirred at 25 °C for 1 h. LCMS showed the starting material was consumed completely. The reaction was diluted with water (15 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give compound PE-AM1-c (24 mg, crude) as a white solid.

[0445] LCMS (ESI): m / z, 715.4 [M+H] + , 737.4 [M+Na] + .

[0446] Step 3: Preparation of compound PE-AM1-d

[0447] To a solution of compound PE-AM1-c (19.44 mg, 27.19 μmol, 1 eq) and 3-N-tert-butoxycarbonylaminocyclobutylamine (5.79 mg, 97%, 32.63 μmol, 1.2 eq) in DMF (1 mL) was added DIPEA (10.65 mg, 99%, 13.62 μL, 81.58 μmol, 3 eq) and HATU (15.99 mg, 97%, 40.79 μmol, 1.5 eq) and stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction was diluted with water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give compound PE-AM1-d (36 mg, crude) as a yellow solid.

[0448] LCMS (ESI): m / z, 869.6 [M+H] + , 891.6 [M+Na] + .

[0449] Step 4: Preparation of compound PE-AM1

[0450] To a solution of compound PE-AM1-d (23.60 mg, 27.16 μmol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.2 mL) at 0 °C and stirred at 0 °C for 2 hours, LCMS showed the reaction was completed, the reaction solution was concentrated and purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, LC-20AP model. The column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) purification, freeze-drying to obtain white solid compound PE-AM1 (2.41 mg, yield 11.54%, purity 98.81%).

[0451] LCMS (ESI): m / z, 769.5 [M+H] + , 791.5 [M+Na] + .

[0452] 1H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 6.19 (d, J = 3.0 Hz, 1H), 5.94 (d, J = 3.0 Hz, 1H), 4.94 (d, J = 2.2 Hz, 1H), 4.88-4.85 (m, 1H), 4.80 (s, 1H), 4.75 (s, 1H), 4.57 (s, 1H), 4.34 (d, J = 6.7 Hz, 1H), 4.29 (d, J = 9.7 Hz, 1H), 4.22 (dd, J = 13.9, 6.6 Hz, 1H), 4.08-4.00 (m, 3H), 3.95 (dd, J = 9.7, 7.2 Hz, 1H), 3.77 (dd, J = 9.8, 2.7 Hz, 1H), 3.71-3.66 (m, 3H), 3.61 (ddd, J = 12.7, 7.8, 4.4 Hz, 3H), 3.54-3.50 (m, 1H), 3.43 (s, 2H), 3.21 (d, J = 6.5 Hz, 4H), 2.97 (dd, J = 9.3, 2.1 Hz, 1H), 2.81-2.73 (m, 2H), 2.66 (tt, J = 4.4, 1.8 Hz, 1H), 2.62-2.54 (m, 3H), 2.44-2.40 (m, 1H), 2.34-2.29 (m, 3H), 2.28-2.23 (m, 1H), 2.11 (ddd, J = 16.1, 6.1, 3.6 Hz, 1H), 2.02-1.89 (m, 3H), 1.87-1.82 (m, 1H), 1.74-1.67 (m, 3H), 1.60 (ddd, J = 17.3, 8.4, 3.6 Hz, 3H), 1.44 (tt, J = 7.7, 4.5 Hz, 2H), 1.34-1.28 (m, 2H), 1.23 (d, J = 3.6 Hz, 2H), 1.03 (d, J = 6.4 Hz, 3H), 0.94 (d, J = 11.9 Hz, 1H).

[0453] Preparation Example 34: Preparation of compound PE-AM2

[0454] Step 1: Preparation of compound PE-AM2-a

[0455] To a solution of compound PE-AM1-c (for the synthesis of this compound, please refer to the preparation of PE-AM1) (16 mg, 22.38 μmol, 1 eq) and tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (5.61 mg, 95%, 26.86 μmol, 1.2 eq) in DMF (1 mL) was added DIPEA (8.77 mg, 99%, 11.21 μL, 67.15 μmol, 3 eq) and HATU (13.16 mg, 97%, 33.57 μmol, 1.5 eq) and stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction solution was added to water (20 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate and rotary evaporated under reduced pressure to give the crude product. The crude product was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) and lyophilized to give white solid compound PE-AM2-a (6 mg, yield 29.96%).

[0456] LCMS (ESI): m / z, 895.4 [M+H] + , 917.4 [M+Na] + .

[0457] Step 2: Preparation of compound PE-AM2

[0458] To a solution of compound PE-AM2-a (6 mg, 6.70 μmol, 1 eq) in dichloromethane (1 mL) was added trifluoroacetic acid (0.2 mL) at 0 °C and stirred at 0 °C for 2 h. LCMS showed the reaction was completed. The reaction solution was concentrated and purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Synergi Max-RP 200 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) and lyophilized to give white solid compound PE-AM2 (1.46 mg, yield 27.40%, purity 95%).

[0459] LCMS (ESI): m / z, 795.4 [M+H] + , 817.4 [M+Na] + .

[0460] 1H NMR (400 MHz, DMSO-d6) δ 6.19 (d, J = 3.0 Hz, 1H), 5.94 (d, J = 3.0 Hz, 1H), 4.95 (s, 1H), 4.87 (s, 1H), 4.80 (s, 1H), 4.76 (s, 1H), 4.57 (s, 1H), 4.33 (s, 1H), 4.29 (d, J = 9.7 Hz, 1H), 4.19 (t, J = 7.9 Hz, 2H), 4.02 (dd, J = 10.1, 6.0 Hz, 2H), 3.91 (s, 2H), 3.77 (d, J = 9.3 Hz, 1H), 3.70 (d, J = 7.1 Hz, 4H), 3.61 (d, J = 6.4 Hz, 2H), 3.51 (s, 2H), 3.43 (d, J = 3.8 Hz, 2H), 3.20 (s, 2H), 2.97 (dd, J = 9.4, 2.1 Hz, 1H), 2.78 (dd, J = 17.3, 6.5 Hz, 2H), 2.67 (p, J = 1.9 Hz, 1H), 2.60 (dd, J = 11.7, 6.5 Hz, 2H), 2.56 (s, 1H), 2.45 - 2.41 (m, 2H), 2.33 - 2.29 (m, 3H), 2.26 (s, 2H), 2.12 (d, J = 8.7 Hz, 1H), 1.95 (s, 2H), 1.85 (d, J = 12.1 Hz, 1H), 1.70 (dd, J = 12.9, 6.3 Hz, 3H), 1.62 (dd, J = 10.0, 5.3 Hz, 3H), 1.43 (s, 2H), 1.31 (t, J = 9.3 Hz, 2H), 1.24 (d, J = 3.5 Hz, 2H), 1.04 (d, J = 6.5 Hz, 3H), 0.95 (d, J = 11.9 Hz, 1H).

[0461] Preparation Example 35: Preparation of compound PE-AM3

[0462] Step 1: Preparation of compound PE-AM3-a

[0463] To a solution of compound PE-AM1-c (20 mg, crude, the synthesis reference to the preparation of PE-AM1) and tert-butyl N-2-azaspiro[3.3]heptan-6-ylcarbamate (6.74 mg, 30.78 μmol, 1.1 eq) in DMF (2 mL) was added HATU (12.77 mg, 33.57 μmol, 1.2 eq) and DIPEA (10.96 mg, 83.93 μmol, 3 eq), the reaction was stirred at room temperature overnight. LCMS showed the reaction was completed. The mixture was prepared by high performance liquid chromatography (preparative chromatography factory Oriendo, model BRIX-2860-R6. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was eluted from 40% to 70%), to get the product PE-AM3-a (11.8 mg, yield 46.39%) as colorless oil, LCMS (ESI): m / z, 909.5 [M+H] + .

[0464] Step 2: Preparation of compound PE-AM3

[0465] To a solution of compound PE-AM3-a (11.8 mg, 12.98 μmol, 1 eq) in DCM (2 mL) was added TFA (0.4 mL), the reaction was stirred at room temperature for half an hour, LCMS showed that the starting material was completely reacted. The mixture was dried by rotary evaporation under reduced pressure to get the crude product, which was prepared by high performance liquid chromatography (preparative chromatography factory Oriendo, model BRIX-2860-R6. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was eluted from 30% to 60%), to get the compound PE-AM3 (3.98 mg, yield 35.74%) as white solid, LCMS (ESI): m / z, 809.5 [M+H] + .

[0466] 1H NMR (400 MHz, DMSO-d6) δ 8.34 (d, J = 2.0 Hz, 1H), 6.19 (d, J = 3.2 Hz, 1H), 5.94 (d, J = 3.2 Hz, 1H), 4.96 - 4.74 (m, 5H), 4.36 - 4.26 (m, 2H), 4.14 - 3.98 (m, 5H), 3.83 - 3.72 (m, 4H), 3.71 - 3.67 (m, 2H), 3.61 - 3.57 (m, 2H), 3.20 (d, J = 1.6 Hz, 4H), 3.00 - 2.94 (m, 1H), 2.81 - 2.72 (m, 2H), 2.69 - 2.53 (m, 4H), 2.46 - 2.20 (m, 9H), 2.16 - 2.07 (m, 1H), 2.02 - 1.81 (m, 6H), 1.75 - 1.55 (m, 6H), 1.43 (d, J = 5.6 Hz, 2H), 1.35 - 1.27 (m, 2H), 1.09 - 0.88 (m, 5H).

[0467] Preparation Example 36: Preparation of compound PE-AM4

[0468] To a solution of compound PE-AM1-c (the synthesis of this compound please refer to the preparation example of PE-AM1) (16 mg, 22.38 μmol, 1 eq) and 3-methoxyazetidine hydrochloride (3.42 mg, 97%, 26.86 μmol, 1.2 eq) in DMF (1 mL) was added DIPEA (8.77 mg, 99%, 11.21 μL, 67.15 μmol, 3 eq) and HATU (10.53 mg, 97%, 26.86 μmol, 1.2 eq) and stirred at 25 °C for 1 hour, LCMS showed the reaction was completed, the reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was YMC-Triart Prep C18 150 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 30% to 60% elution), and was freeze-dried to obtain white solid compound PE-AM4 (11.14 mg, yield 63.49%).

[0469] LCMS (ESI): m / z, 784.4 [M+H] + , 806.4 [M+Na] + .

[0470] 1H NMR (400 MHz, DMSO-d6) δ 5.11 - 4.96 (m, 2H), 4.90 - 4.70 (m, 3H), 4.63 (dt, J = 4.6, 2.6 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.17 (dt, J = 10.2, 5.0 Hz, 1H), 4.14 - 4.06 (m, 4H), 4.05 - 3.96 (m, 2H), 3.84 - 3.75 (m, 3H), 3.70 (ddd, J = 9.5, 5.7, 2.8 Hz, 1H), 3.51 (td, J = 11.5, 10.4, 4.8 Hz, 5H), 3.31 (t, J = 3.9 Hz, 1H), 3.26 (s, 3H), 2.86 - 2.71 (m, 3H), 2.64 - 2.53 (m, 2H), 2.36 - 2.18 (m, 7H), 2.12 (t, J = 11.9 Hz, 1H), 2.04 - 1.86 (m, 6H), 1.74 - 1.59 (m, 4H), 1.54 - 1.41 (m, 2H), 1.35 - 1.13 (m, 4H), 1.03 (d, J = 6.4 Hz, 3H), 1.00 - 0.91 (m, 1H).

[0471] Preparation Example 37: Preparation of compound PE-AM5

[0472] To a solution of compound PE-AM1-c (the synthesis of this compound please refer to the preparation example of PE-AM1) (16 mg, 22.38 μmol, 1 eq) and 5-amino-1-pentanol (2.92 mg, 95%, 26.86 μmol, 1.2 eq) in DMF (1 mL) was added DIPEA (8.77 mg, 99%, 11.21 μL, 67.15 μmol, 3 eq) and HATU (13.16 mg, 97%, 33.57 μmol, 1.5 eq) and stirred at 25 °C for 1 hour, LCMS showed the reaction was completed, the reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, acetonitrile was eluted from 20% to 50% elution) and lyophilized to obtain white solid compound PE-AM5 (10.86 mg, yield 60.65%).

[0473] LCMS (ESI): m / z, 800.5 [M+H] + , 822.4 [M+Na] + .

[0474] 1 H NMR (400 MHz, DMSO-d6) δ 7.78 (t, J = 5.6 Hz, 1H), 5.09 - 4.99 (m, 2H), 4.78 (d, J = 25.4 Hz, 2H), 4.63 (dt, J = 4.6, 2.5 Hz, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.34 (t, J = 5.1 Hz, 1H), 4.26 (d, J = 10.9 Hz, 1H), 4.17 (td, J = 11.0, 10.6, 4.5 Hz, 1H), 4.10 (t, J = 3.3 Hz, 2H), 3.99 (dt, J = 8.2, 4.8 Hz, 2H), 3.79 (t, J = 10.4 Hz, 1H), 3.68 (s, 1H), 3.56 - 3.47 (m, 2H), 3.38 (s, 2H), 3.29 (d, J = 3.5 Hz, 1H), 3.25 (s, 2H), 3.00 (dp, J = 19.6, 6.3 Hz, 2H), 2.85 - 2.72 (m, 3H), 2.56 (dd, J = 16.6, 10.0 Hz, 1H), 2.43 - 2.37 (m, 1H), 2.35 - 2.19 (m, 6H), 2.12 (t, J = 11.9 Hz, 1H), 2.01 - 1.84 (m, 6H), 1.67 (q, J = 11.6 Hz, 4H), 1.56 - 1.46 (m, 2H), 1.40 - 1.34 (m, 4H), 1.29 - 1.22 (m, 4H), 1.20 - 1.12 (m, 1H), 1.03 (d, J = 6.4 Hz, 3H), 0.94 (d, J = 6.6 Hz, 1H), 0.87 - 0.81 (m, 3H).

[0475] Preparation Example 38: Preparation of compound PE-O2K

[0476] Step 1: Preparation of compound PE-O2K-b

[0477] To a solution of compound PE-AM1-a (purchased from Haoyuan Pharmaceutical) (150 mg, 0.205 mmol, 1 eq) in DMF (3 mL) was added compound TBDPSCl (378 mg, 1.373 mmol, 6.7 eq) and imidazole (97.8 mg, 1.437 mmol, 7 eq), after addition, 25 °C was stirred for 1 hour. LCMS showed that the reaction was complete. The reaction liquid was filtered, the filtrate was added with water (30 mL), then extracted with EtOAc (50 mL x 3), the organic phase was combined and washed with brine (30 mL), dried over anhydrous sodium sulfate, after filtration, the filtrate was obtained, the filtrate was concentrated under reduced pressure to obtain the crude product, the residue was purified by reversed phase column (ACN: H2O = 0: 100-100: 0) to obtain the yellow liquid product (98.0 mg, yield 49%).

[0478] LCMS (ESI): m / z, 991.3 [M+Na] + , Rt = 2.075 min.

[0479] Step 2: Preparation of compound PE-O2K-b1

[0480] To a solution of compound PE-O2K-b (98 mg, 0.101 mmol, 1 eq) in THF (4 mL) and water (1 mL) was added potassium osmate dihydrate (4 mg, 0.010 mmol, 0.1 eq) and NaIO4(433 mg, 2.022 mmol, 20 eq) at 0 °C, the reaction liquid was stirred at 25 °C for 2 hours. NaIO4(433 mg, 2.022 mmol, 20 eq) was added to the reaction, the reaction liquid was stirred at room temperature overnight, LCMS showed that the reaction was complete. The reaction liquid was filtered, the filtrate was added with water (30 mL), then extracted with EtOAc (50 mL x 3), the organic phase was combined and washed with brine (30 mL), dried over anhydrous sodium sulfate, after filtration, the filtrate was obtained, the filtrate was concentrated under reduced pressure to obtain the crude product yellow liquid product PE-O2K-b1 (130 mg, crude).

[0481] LCMS (ESI): m / z, 994.7 [M+Na] + , Rt = 1.822 min.

[0482] Step 3: Preparation of compound PE-O2K

[0483] To a solution of compound PE-O2K-b1 (90.0 mg, 0.092 mmol, 1 eq) in DMF (4 mL) was added triethylamine trihydrofluoride (0.2 mL) at room temperature. The reaction was stirred at 65 °C for 2.5 hours. LCMS showed the starting material almost reacted completely. The reaction was concentrated, and the residue was prepared by high performance liquid chromatography to give the product PE-O2K (9.28 mg, yield 14%) as a white solid.

[0484] The high performance liquid chromatography preparation conditions were as follows:

[0485] The preparation chromatography manufacturer was Shimadzu, model JB-C235-04. The chromatographic column was Prime C18 30 x 150 mm x 5 μm. The mobile phase was water (0.1% TFA) - ACN, and the elution ratio of acetonitrile was eluted from 23% to 43%.

[0486] LCMS (ESI): m / z, 735.2 [M+H] + , Rt = 1.140 min.

[0487] 1 H NMR (400 MHz, DMSO-d6) δ 4.65 (t, J = 4.4 Hz, 1H), 4.56 (d, J = 4.2 Hz, 1H), 4.39 (s, 1H), 4.12 (dd, J = 12.0, 10.6 Hz, 4H), 3.86 (d, J = 8.8 Hz, 1H), 3.77 (ddd, J = 14.2, 9.4, 5.2 Hz, 4H), 3.56 - 3.48 (m, 2H), 3.26 (d, J = 7.8 Hz, 8H), 2.85 (ddd, J = 13.8, 8.0, 5.1 Hz, 2H), 2.79 - 2.72 (m, 1H), 2.68 - 2.53 (m, 2H), 2.39 - 1.89 (m, 10H), 1.74 (ddd, J = 14.6, 12.2, 8.2 Hz, 4H), 1.67 - 1.38 (m, 8H), 1.31 - 1.20 (m, 2H), 0.94 (d, J = 6.6 Hz, 2H).

[0488] Preparation Example 39: Preparation of compound PY-4car2-b

[0489] To a solution of 4-((tert-butyldimethylsilyl)oxy)butan-1-ol (1 g, 4.89 mmol, 1 eq) and bis(p-nitrophenyl) carbonate (2.98 g, 9.79 mmol, 2 eq) in N,N-dimethylformamide (15 mL) was added N,N-diisopropylethylamine (2.43 mL, 14.68 mmol, 3 eq) and stirred at room temperature overnight. The reaction was concentrated, diluted with dichloromethane (10 mL), washed with water (10 mL) for 3 times, washed with saturated brine (10 mL) for 1 time, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1:10, dichloromethane / petroleum ether = 40%) to give the product 4-((tert-butyldimethylsilyl)oxy)butyl (4-nitrophenyl) carbonate (PY-4car2-b) (1.63 g, yield = 90.2%).

[0490] Preparation Example 40: Preparation of compound LY-46F-c

[0491] Step 1: Preparation of (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-(4- (hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2- yl)carbamate

[0492] To a solution of compound (((9H-fluoren-9-yl)methoxy)carbonyl)-L-valine-L-alanine (1 g, 2.44 mmol, 1 eq) in DCM (20 mL) / MeOH (10 mL) was added (4- aminophenyl)methanol (360.04 mg, 2.92 mmol, 1.2 eq) and EEDQ (2-ethoxy-1- ethoxy carbonyl-1,2-dihydroquinoline, 903.71 mg, 753.09 μL, 3.65 mmol, 1.5 eq). The reaction was stirred at 25 °C for 12 h. The reaction was concentrated under reduced pressure, the obtained solid was slurry with petroleum ether / ethyl acetate = 3:1, total 30 mL, the filter cake was washed with petroleum ether / ethyl acetate = 3:1, total 50 mL to give the product (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-(4- (hydroxymethyl)phenyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2- yl)carbamate (1.10 g, yield 87.57%) as a yellow solid.

[0493] LCMS (ESI): m / z, 538.2 [M+Na] + .

[0494] Step 2: Preparation of compound LY-46F-c

[0495] (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-(4-(hydroxymethyl)phenyl)amino)-1- oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)carbamate (500 mg, 969.74 pmol, 1 eq) was dissolved in DMF (6 mL), to the solution was added bis(4-nitrophenyl) carbonate (590.01 mg, 393.34 pL, 1.94 mmol, 2 eq) and DIPEA (375.99 mg, 480.80 pL, 2.91 mmol, 3 eq). The reaction was stirred at 25 °C for 16 h. The reaction was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether 1:30 to 1:1) to give compound LY-46F-c (380 mg, yield 57.57%) as a yellow solid.

[0496] LCMS (ESI): m / z, 703.3 [M+Na] + .

[0497] Preparation Example 41: Preparation of compound LY-22CD-3-d

[0498] Step 1: Preparation of 5-bromothiazolo[5,4-b]pyridin-2-amine

[0499] To a solution of 5-amino-2-bromopyridine (10 g, 57.80 mmol, 1 eq) and potassium thiocyanate (22.47 g, 11.91 mL, 231.20 mmol, 4 eq) in acetic acid (100 mL) was added a solution of bromine in acetic acid (20 mL) at 0 °C, and stirred at 20 °C for 3 h. The reaction was quenched with water (50 mL), and the filtrate was obtained by filtration. The filtrate was concentrated under reduced pressure to give a crude product. The crude product was basified with sodium bicarbonate aqueous solution to pH = 8, and extracted with ethyl acetate (50 mL x 4), and the organic phase was combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure to give 5-bromothiazolo[5,4-b]pyridin-2-amine (9.80 g) as a yellow solid.

[0500] LCMS (ESI): m / z, 230.0 [M+H] + .

[0501] 1 H NMR (400 MHz, DMSO-d 6 ) d 7.96 (s, 2H), 7.57 (d, J = 8.4 Hz, 1H), 7.43 (d, J = 8.4 Hz, 1H).

[0502] Step 2: Preparation of tert-butyl (5-bromothiazolo[5,4-b]pyridin-2-yl)carbamate

[0503] To a solution of 5-bromothiazolo[5,4-b]pyridine-2-carboxylic acid (2.00 g, 8.62 mmol, 1 eq) in tetrahydrofuran (80 mL) was added 2-mercaptoacetic acid (0.80 g, 8.62 mmol, 1 eq) at room temperature. The mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated, and the residue was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 20%-30%) to give 5-bromothiazolo[5,4-b]pyridine-2-carbothioic acid (1.50 g, 6.62 mmol, 77% yield) as a yellow solid.

[0504] LCMS (ESI): m / z, 330.0 [M+H] + .

[0505] 1 1H NMR (400 MHz, DMSO-d6) d 12.12 (s, 1H), 7.99 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 8.4 Hz, 1H), 1.52 (s, 9H).

[0506] Step 3: Preparation of methyl 2-((tert-butoxycarbonyl)amino)thiazolo[5,4- b]pyridine-5-carboxylate

[0507] To a solution of tert-butyl (5-bromothiazolo[5,4-b]pyridin-2-yl)carbamate (6 g, 18.17 mmol, 1 eq), potassium carbonate (7.53 g, 3.10 mL, 54.51 mmol, 3 eq), palladium acetate (1.22 g, 5.45 mmol, 0.3 eq) and 1,3-bisdiphenylphosphinopropane (1.80 g, 4.36 mmol, 0.24 eq) in methanol (120 mL) and N,N-dimethylformamide (60 mL) was added under carbon monoxide atmosphere at room temperature. The mixture was stirred at 60 °C for 16 h. The reaction mixture was concentrated, and the residue was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 40%-80%) to give methyl 2-((tert-butoxycarbonyl)amino)thiazolo[5,4-b]pyridine-5-carboxylate (2.20 g, 7.11 mmol, 39% yield).

[0508] LCMS (ESI): m / z, 310.1 [M+H] + .

[0509] 1H NMR (400 MHz, DMSO-d6) δ 12.26 (s, 1H), 8.25-8.04 (m, 2H), 3.90 (s, 3H), 1.53 (s, 9H).

[0510] Step 4: Preparation of methyl 2-aminothiazolo[5,4-b]pyridine-5-carboxylate

[0511] To a solution of methyl 2-((tert-butoxycarbonyl)amino)thiazolo[5,4-b]pyridine-5- carboxylate (2.20 g, 7.11 mmol, 1 eq) in dichloromethane (80 mL) was added trifluoroacetic acid (40 mL) at 0 °C, and stirred at 20 °C for 2 h. The reaction was concentrated, and then the residue was added to saturated aqueous sodium bicarbonate solution, and then filtered, and the filter cake was concentrated under reduced pressure to give methyl 2-aminothiazolo[5,4-b]pyridine-5-carboxylate (1.38 g, 6.60 mmol, 93% yield).

[0512] LCMS (ESI): m / z, 210.1 [M+H] + .

[0513] 1 H NMR (400 MHz, Chloroform-d) δ 8.37-8.27 (m, 2H), 4.07 (s, 3H).

[0514] Step 5: Preparation of methyl 2-bromothiazolo[5,4-b]pyridine-5-carboxylate

[0515] To a solution of copper bromide (1.84 g, 386.06 μL, 8.24 mmol, 1.25 eq) in acetonitrile (60 mL) was added tert-butyl nitrite (1.22 g, 1.42 mL, 11.87 mmol, 1.8 eq), methyl 2- aminothiazolo[5,4-b]pyridine-5-carboxylate (1.38 g, 6.60 mmol, 1 eq) at 0 °C, and stirred at 25 °C for 16 h. The reaction was concentrated, and then the residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 20%-40%) to give the product methyl 2-bromothiazolo[5,4-b]pyridine-5-carboxylate (1.45 g, 5.31 mmol, 81% yield).

[0516] LCMS (ESI): m / z, 273.1 [M+H] + .

[0517] 1 H NMR (400 MHz, Chloroform-d) δ 8.37-8.27 (m, 2H), 4.07 (s, 3H).

[0518] Step 6: Preparation of methyl 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylate

[0519] A solution of methyl 2-bromo-[l,3]thiazolo[5,4-b]pyridine-5-carboxylate (375 mg, 1.37 mmol, 1 eq) and sodium methanethiolate (240.61 mg, 3.43 mmol, 2.5 eq) in ethyleneglycol dimethyl ether (10 mL) was stirred at -15 °C for 2 h. To the reaction was added saturated aqueous ammonium chloride solution (5 mL), concentrated, added tetrahydrofuran (30 mL), filtered, collected the filter cake, purified by high performance liquid preparation chromatography (Oriendo, Model BRIX-2860-R1. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give methyl 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylate (180 mg, yield 25.89%) and 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylic acid (100 mg, yield 15.28%) as white solids.

[0520] LCMS (ESI): m / z, 241.1 [M+H] + .

[0521] Step 7: Preparation of 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylic acid

[0522] A mixture of methyl 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylate (180 mg, 749.06 pmol, 1 eq) and lithium hydroxide monohydrate (62.92 mg, 41.67 pL, 1.50 mmol, 2 eq) in tetrahydrofuran (2 mL) and water (2 mL) was stirred at room temperature for 12 h at 20 °C. The reaction was concentrated, to which was added water (50 mL) and aqueous hydrochloric acid (2 M) to pH < 5, filtered, washed with water (3 mL x 2), and the filter cake was collected to give the product 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylic acid (169 mg, crude).

[0523] LCMS (ESI): m / z, 227.1 [M+H] + .

[0524] Step 8: Preparation of 2-methylsulfonyl-thiazolo[5,4-b]pyridine-5-carboxylic acid

[0525] To a solution of 2-(methylthio)thiazolo[5,4-b]pyridine-5-carboxylic acid (269 mg, 1.19 mmol, 1 eq) in tetrahydrofuran (10 mL) and water (10 mL) was added potassium peroxymonosulfate (3.29 g, 9.51 mmol, 8 eq), the mixture was stirred at 20 °C for 2 h. To the mixture was added ethyl acetate (30 mL) and H2O (20 mL), the aqueous phase was extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with saturated brine (10 mL) and dried over anhydrous sodium sulfate, filtered and concentrated, the residue was purified by high performance liquid preparation chromatography (Preparation Chromatography Factory Oriendo, Model BRIX-2860. The column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 10% to 40% elution) to give the product 2-methylsulfonyl-thiazolo[5,4-b]pyridine-5-carboxylic acid (220 mg, yield 71.65%).

[0526] LCMS (ESI): m / z, 259.1 [M+H] + .

[0527] Step 9: Preparation of compound LY-22CD-3-a

[0528] To a solution of tert-butyl 3-(2-aminoethoxy)propanoate (70.34 mg, 371.70 pmol, 1.2 eq) and 2-methylsulfonyl-thiazolo[5,4-b]pyridine-5-carboxylic acid (80 mg, 309.75 pmol, 1 eq) in N,N-dimethylformamide (4 mL) was added N,N-diisopropylethylamine (80.07 mg, 102.38 pL, 619.51 pmol, 2 eq) and N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-l-yl)uronium hexafluorophosphate (141.34 mg, 371.70 pmol, 1.2 eq), stirred at room temperature for 1 h. The reaction solution was filtered and purified by high performance liquid preparation chromatography (Preparation Chromatography Factory Oriendo, Model BRIX-2860-R6. The column was Synergi Max-RP 250 x 40 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give compound LY-22CD-3-a (114.3 mg, yield 85.91%).

[0529] LCMS (ESI): m / z, 452 [M+Na] + .

[0530] Step 10: Preparation of compound LY-22CD-3-b

[0531] To a solution of compound LY-22CD-3-a (114.30 mg, 266.12 pmol, 1 eq) in dichloromethane (5 mL) was added trifluoroacetic acid (1 mL) under ice-bath, then stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give the crude product (150 mg).

[0532] LCMS (ESI): m / z, 396 [M+Na] + .

[0533] Step 11: Preparation of compound LY-22CD-3-c

[0534] To a solution of compound LY-22CD-3-b (99.37 mg, 266.12 pmol, 1 eq) in N,N- dimethylformamide (4 mL) was added N-hydroxysuccinimide (183.77 mg, 1.60 mmol, 6 eq) and N,N'-diisopropylcarbodiimide (201.51 mg, 248.77 pL, 1.60 mmol, 6 eq) under ice-bath, stirred at 50 °C for 2 hours. The reaction mixture was filtered, then purified by high performance liquid preparative chromatography (Oriendo, BRIX-2860. Column: YMC-Triart Prep C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 10% to 40% elution) to give compound LY-22CD-3-c (100 mg, yield 79.87%).

[0535] LCMS (ESI): m / z, 493 [M+Na] + .

[0536] Step 12: Preparation of compound LY-22CD-3-d

[0537] To a solution of compound LY-22CD-3-c (100 mg, 212.55 pmol, 1 eq) and 27-amino-4,7,10,13,16,19,22,25-octaoxahexacosanoic acid (112.61 mg, 102.37 pL, 255.06 pmol, 1.2 eq) in N,N-dimethylformamide (3 mL) was added N,N-diisopropylethylamine (54.94 mg, 70.26 pL, 425.11 pmol, 2 eq) and stirred at room temperature for 1 h. The reaction was filtered and purified by high performance liquid chromatography (preparative chromatography, Shimadzu, LC-20AP. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution from 10% to 40%) to give compound LY-22CD-3-d (48.4 mg, yield 28.57%).

[0538] LCMS (ESI): m / z, 797 [M+H] + .

[0539] Preparation Example 42: Preparation of compound LY-5a

[0540] N 6 -(tert-butoxycarbonyl)-N 2 Preparation of (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)-L-lysine (LY-5a)

[0541] Into a reaction flask was added N 6 -(tert-butoxycarbonyl)-L-lysine (300 mg, 1.0 eq), 2,5-dioxopyrrolidin-1-yl 6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoate (Ae) (375 mg, 1.0 eq), DIPEA (315 mg, 2 eq) and 10 mL of anhydrous DMF under nitrogen atmosphere. After three times of nitrogen replacement, the reaction was stirred at room temperature for 16 h. Separation by high performance liquid chromatography (Oriendo, BRIX-2860; Column: Phenomenex Luna C18 250 x 50 mm x 10 pm; Mobile phase: water (0.225% trifluoroacetic acid) - acetonitrile, elution from 45% to 85%) to give 400 mg of compound LY-5a as a colorless gum.

[0542] LCMS (ESI): m / z, 340.2 [M-100+1] + , 462.3 [M+Na] + .

[0543] Preparation 43: Preparation of compound LY-82

[0544] Preparation of (18S, 19R, 20R, 21R)-18, 19, 20, 21, 22-pentahydroxy-16-(2S, 3R, 4R, 5R)- 2, 3, 4, 5, 6-pentahydroxyhexyl)-4, 7, 10, 13-tetraoxa-16-azadocosanoic acid (LY-82)

[0545] To a solution of 1-amino-3, 6, 9, 12-tetraoxaheptadecan-15-oic acid (LY-81) (200 mg, 0.75 mmol, 1.0 eq) in anhydrous MeOH (15 mL) and HOAc (1.5 mL) was added (2R, 3S, 4R, 5R)-2, 3, 4, 5, 6-pentahydroxyhexanal (D-glucose) (1.36 g, 7.54 mmol, 10 eq) and NaBH3CN (237 mg, 3.77 mmol, 5 eq) and stirred at 40-45 °C for 24 h under nitrogen atmosphere. The reaction was concentrated under reduced pressure, the residue was taken in ~8 mL water and 2 mL methanol and purified by preparative high performance liquid chromatography (Oriendo, BRIX-2860; column: Phenomenex Luna C18 250 x 50 mm x 10 pm; mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile ratio from 45% to 85% elution) and lyophilized to get compound LY-82 (384 mg, yield 72%, TFA salt) (colorless / whiteish gum).

[0546] LCMS (ESI): m / z, 594.5 [M+1] + .

[0547] Preparation 44: Preparation of compound LY-27a

[0548] Step 1: Preparation of compound LY-27a2

[0549] To a solution of (((9H-fluoren-9-yl)methoxy)carbonyl)-L-valine-L-alanine (Haoyuan Pharmaceutical, LY-27al, 8.5 g, 20.71 mmol, 1 eq) and glycine tert-butyl ester (3.26 g, 24.85 mmol, 1.2 eq) in methanol / dichloromethane (50 mL / 100 mL) was added 2-ethoxy-1-ethoxycarbonyl-1, 2-dihydroquinoline (7.68 g, 31.06 mmol, 1.5 eq) under ice bath, stirred for 10 min and then stirred at 40 °C overnight. After concentration, the residue was slurry in petroleum ether / ethyl acetate (100 mL / 50 mL), stirred for 20 min and then filtered to get white solid (10.6 g, yield 97.76%).

[0550] LCMS (ESI): m / z, 546 [M+Na] + .

[0551] Step 2: Preparation of compound LY-27a3

[0552] To a solution of compound LY-27a2 (10.6 g, 20.24 mmol, 1.0 eq) in dichloromethane (60 mL) was added trifluoroacetic acid (20 mL) under ice-bath, and the reaction was allowed to warm to room temperature and stirred overnight. Concentration and concentration with dichloromethane (10 mL) for 3 times gave the crude product as a light yellow thick paste (15.24 g).

[0553] LCMS (ESI): m / z, 468 [M+H] + .

[0554] Step 3: Preparation of compound LY-27a

[0555] To a solution of compound LY-27a3 (15.24 g, 20.2 mmol, 1.0 eq, 62.1% purity) in N,N-dimethylformamide (100 mL) was added lead tetraacetate (23.3 g, 52.52 mmol, 2.6 eq), acetic acid (4.6 mL, 80.8 mmol, 4 eq), copper acetate (1.46 g, 8.08 mmol, 0.4 eq) under nitrogen atmosphere. The reaction was stirred at 60 °C for 1 hour, then allowed to warm to room temperature, concentrated under reduced pressure, and the residue was taken up in water to give a large amount of insoluble material, which was dissolved in water (100 mL) and acetonitrile (100 mL), extracted with ethyl acetate (100 mL), washed with brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound LY-27a (8 g, 82.2% yield, 80% purity).

[0556] LCMS (ESI): m / z, 504 [M+Na] + .

[0557] 1 H NMR (400 MHz, Methanol-d4) δ 7.81 (d, J = 7.6 Hz, 2H), 7.69 (t, J = 7.0 Hz, 2H), 7.40 (t, J = 7.6 Hz, 2H), 7.32 (t, J = 7.4 Hz, 2H), 5.20 (d, J = 6.2 Hz, 2H), 4.48 - 4.30 (m, 3H), 4.24 (t, J = 6.8 Hz, 1H), 3.94 (d, J = 6.8 Hz, 1H), 2.01 (s, 3H), 1.36 (d, J = 7.2 Hz, 3H), 0.97 (dd, J = 9.6, 6.6 Hz, 6H).

[0558] Preparation 45: Preparation of compound LY-22CC-c

[0559] Step 1: Preparation of compound LY-22CC-a

[0560] Compound LY-27a (500 mg, 1.04 mmol, 1 eq) was dissolved in THF (10 mL), then added compound 4-((tert-butyldimethylsilyl)oxy)butan-1-ol (424.42 mg, 2.08 mmol, 2 eq) and TsOH (p-toluenesulfonic acid, 16.46 mg, 95.59 μmol, 1 eq), the reaction was stirred at 30 °C for 4 hours. The reaction was directly used for the next step.

[0561] LCMS (ESI): m / z, 648.4 [M+Na] + .

[0562] Step 2: Preparation of compound LY-22CC-b

[0563] Compound LY-22CC-a (500 mg, 798.88 μmol, 1 eq) was dissolved in THF (10 mL), to the solution was added 4M hydrochloric acid 1,4-dioxane solution (29.13 mg, 199.72 μL, 798.88 μmol, 1 eq), the solution was stirred at 15 °C for 3 hours. The reaction was concentrated under reduced pressure, the residue was purified by high pressure liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was XT-15 x 30-10. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 40% to 70% elution) to give white solid compound LY-22CC-b (70 mg, yield 17.13%).

[0564] LCMS (ESI): m / z, 534.3 [M+Na] + .

[0565] Step 3: Preparation of compound LY-22CC-c

[0566] Compound LY-22CC-b (70 mg, 136.82 μmol, 1 eq) was dissolved in DMF (5 mL), bis(4-nitrophenyl) carbonate (83.24 mg, 55.50 μL, 273.64 μmol, 2 eq) and DIPEA (53.05 mg, 67.84 μL, 410.46 μmol, 3 eq) were added. The reaction was stirred at 15 °C for 12 h. The reaction was concentrated most of DMF, DCM (30 mL) was added, washed with water (30 mL), the organic phase was dried, filtered, concentrated under reduced pressure, the residue was purified by silica gel column chromatography (eluent: DCM:MeOH = 0:1 to 5:1) to give the product LY-22CC-c (70 mg, yield 75.60%) as a yellow solid.

[0567] LCMS (ESI): m / z, 699.3 [M+Na] + .

[0568] Preparation Example 46: Preparation of compound LY-29C-c

[0569] Step 1: Preparation of ((1R,8S,9s)-bicyclo[6.1.0]non-4-yn-9-yl)methyl (4- nitrophenyl) carbonate (LY-29C-b)

[0570] To compound (1R,8S,9S)-bicyclo[6.1.0]non-4-yn-9-ylmethanol (LY-29C-a) (80 mg, 532.55 μmol, 1 eq, Shengdong Xingzhi Bio) in anhydrous DCM (5 mL) under nitrogen atmosphere, pyridine (105.31 mg, 107.68 μL, 1.33 mmol, 2.5 eq) and 4-nitrophenyl chloroformate (134.18 mg, 665.69 μmol, 1.25 eq) were added at 0 °C, and the reaction was allowed to react at room temperature for half an hour. The reaction was quenched with saturated ammonium chloride (10 mL) at 0 °C, extracted with DCM (40 mL x 3) three times, the organic phase was washed with brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:9) to give compound LY-29C-b (75 mg, yield 44.66%) as anhydrous oil.

[0571] Step 2: Preparation of compound LY-29C-c

[0572] To a mixture of compound LY-29B-a (156.82 mg, 355.18 μmol, 1 eq) and DIPEA (114.77 mg, 146.76 μL, 887.96 μmol, 2.5 eq) in anhydrous DMF (5 mL) was added LY-29C-b (112 mg, 355.18 μmol, 1 eq) at 0 °C under nitrogen atmosphere. The reaction was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure at no more than 40 °C. The residue was purified by high performance liquid preparation chromatography (preparative chromatography manufacturer: ISCO, model: ISCO-R1. Column: Synergi Max-RP 250 x 40 mm x 10 μm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 21% to 51% elution) to give compound LY-29C-c (148 mg, yield 67.45%) as anhydrous oil.

[0573] Example

[0574] Example 1: Preparation of compound BA-1

[0575] Step 1: Preparation of compound BA-1-b

[0576] To a mixture of compound BA-1-a (purchased from Bide) (40.68 mg, 158.12 μmol, 1.2 eq) and compound LY-3a (purchased from Bide) (50 mg, 131.77 μmol, 1 eq) in THF (1 mL) and MeOH (1 mL) was added DMTMM (4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride) (87.51 mg, 316.25 μmol, 2.4 eq) at 0 °C. The reaction was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction mixture was concentrated to dryness. The crude product was purified by trituration (acetonitrile 3 mL) to give compound BA-1-b (75 mg, yield 92%) as a white solid.

[0577] LCMS (ESI): m / z, 619.4 [M+H] + Rt = 1.530 min.

[0578] Step 2: Preparation of compound BA-1-c

[0579] To a solution of compound BA-1-b (75 mg, 121.22 pmol, 1 eq) in DMF (5 mL) was added DIPEA (47 mg, 60.10 pL, 363.67 pmol, 3 eq) and NPC (184.39 mg, 122.93 pL, 606.12 pmol, 5 eq) at 0 °C. The reaction was stirred at 33 °C for 2 h. LCMS showed the reaction was completed. The reaction was concentrated to get the crude product. The crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane 1 :20 to 1 :10) to give compound BA-1-c (83 mg, yield 87.36%) as a white solid.

[0580] LCMS (ESI): m / z, 784.5 [M+H] + Rt = 1.778 min.

[0581] Step 3: Preparation of compound BA-1

[0582] To a solution of compound BA-1-c (12.34 mg, 15.74 pmol, 1.3 eq) and eribulin mesylate (10 mg, 12.11 pmol, 1 eq) in DMF (3 mL) was added DIPEA (4.69 mg, 6 pL, 36.32 pmol, 3 eq) at 0 °C. The reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was purified by high performance liquid chromatography preparation (Prep-chromatography manufacturer Oriendo, model BRIX-2860. The column was Synergi Max-RP 200 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of water was from 30% to 60% elution) and lyophilized to give compound BA-1 (2130869-18-8, 10.38 mg, yield 62.37%, purity 98.24%) as a white solid.

[0583] LCMS (ESI): m / z, 1374.9 [M+H] + , 1397.9 [M+Na] + Rt = 2.213 min.

[0584] Example 2: Preparation of compound BA-2

[0585] To a solution of compound eribulin mesylate (3 mg, 3.63 pmol, 1 eq) and BA-2-a (4.02 mg, 5.45 pmol, 1.5 eq) in DMF (1 mL) was added DIPEA (1.41 mg, 1.80 pL, 10.90 pmol, 3 eq) at 0 °C. After addition, the mixture was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction mixture was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The column was Welch Xtimate C18 150 x 21.2 mm x 5 pm. The mobile phase was water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 40% to 70% elution) and lyophilized to give compound LA-B2 (2.39 mg, yield 49.53%, purity 99%) as a white solid.

[0586] LCMS (ESI): m / z, 1351.6 [M+Na] + Rt = 2.222 min.

[0587] Example 3: Preparation of compound BA-3

[0588] Step 1: Preparation of compound BA-3-c

[0589] To a solution of compound LY-46F-c (10.42 mg, 87%, 13.32 pmol, 1.1 eq) and eribulin mesylate (10 mg, 12.11 pmol, 1 eq) in DMF (1 mL) was added DIPEA (4.69 mg, 6 pL, 36.32 pmol, 3 eq) and HOBT (1.64 mg, 12.11 pmol, 1 eq). After the mixture was stirred at room temperature for 2 h, LCMS showed the starting material was consumed. The reaction mixture was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer ISCO, model ISCO. The column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 60% to 90% elution) to give product BA-3-c (5.50 mg, yield 35.73%) as a white solid. LCMS (ESI): m / z, 1293 [M+Na] + .

[0590] Step 2: Preparation of compound BA-3-d

[0591] To a solution of compound BA-3-c (5.50 mg, 4.33 μmol, 1 eq) in DMF (1 mL) was added DBU (1.98 mg, 1.94 μL, 12.98 μmol, 3 eq), after reaction at room temperature for 40 min, LCMS showed the starting material disappeared, TsOH (2.23 mg, 12.98 μmol, 3 eq) was added to quench the reaction, then used directly for next step. LCMS (ESI): m / z, 1071 [M+Na] + .

[0592] Step 3: Preparation of compound BA-3

[0593] To a solution of compound BA-3-d (3.45 mg, 4.33 μmol, 1 eq), LY-22CD-3-d (3.45 mg, 4.33 μmol, 1 eq) in DMF (1 mL) was added DIPEA (1.12 mg, 1.43 μL, 8.65 μmol, 2 eq) and HATU (1.97 mg, 5.19 μmol, 1.2 eq) at 0 °C, after reaction at room temperature for 1 h, LCMS showed the starting material disappeared, purified by high performance liquid chromatography-preparation (preparative chromatography manufacturer ISCO, model ISCO. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give the product BA-3 (3.45 mg, yield 43.62%, purity 97%) as a white solid, LCMS (ESI): m / z, 1849 [M+Na] + .

[0594] Example 4: Preparation of compound BA-4

[0595] Step 1: Preparation of compound BA-4-a

[0596] To a solution of compound BA-3-d (38 mg, 36.22 pmol, 1 eq, the synthesis of this compound is referred to the synthesis of BA-3) and 2-[(2S)-6-{[(tert-butoxy)carbonyl]amino}-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)hexanamido]acetic acid (21.15 mg, 39.84 pmol, 1.1 eq, the synthesis of this compound is referred to patent application WO2024175069A1) in DME (1.5 mL) was added DIPEA (12.09 pL, 72.43 pmol, 2 eq) and HATU (16.52 mg, 43.46 pmol, 1.2 eq) at 0 °C and stirred at 25 °C for 1 h, LCMS showed the starting material was consumed completely. The residue was purified by high performance liquid chromatography preparation (Prep-Chromatography Shimadzu, LC-20AP. The column was Synergi Max-RP 250 x 40 mm x 10 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, the elution ratio of acetonitrile was from 50% to 80% elution) to give BA-4-a (29.60 mg, yield 52.50%) as a white solid.

[0597] LCMS (ESI): m / z, 1578 [M+Na] + Rt = 3.068 min.

[0598] Step 2: Preparation of compound BA-4-b

[0599] To a solution of compound BA-4-a (29.6 mg, 18.8 pmol, 1 eq) in DCM (3 mL) was added TFA (0.5 mL) at 0 °C, stirred for 30 min, LCMS showed the starting material was consumed completely, concentrated and directly used for next step. The theoretical mass of BA-4-b was 27 mg.

[0600] LCMS (ESI): m / z, 1478 [M+Na] + Rt = 2.443 min.

[0601] Step 3: Preparation of compound BA-4-c

[0602] To a solution of compound BA-4-b (18 mg, 12.36 pmol, 1 eq) and LY-10b (11.41 mg, 14.83 pmol, 1.2 eq, the synthesis of this compound is referred to the preparation of LA-27C-2) in DMF (1 mL) was added DIPEA (12.25 pL, 74.14 pmol, 6 eq) and HATU (5.64 mg, 14.83 pmol, 1.2 eq) at 0 °C and stirred at 25 °C for 1 h, LCMS showed the starting material was consumed. The two batches of reaction solution with the same scale were combined and purified by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was eluted from 30-60%) to give BA-4-c (35.60 mg, yield 66%) as a white solid.

[0603] LCMS (ESI): m / z, 1126 [1 / 2 M+Na] + Rt = 2.262 min.

[0604] Step 4: Preparation of compound BA-4-d

[0605] To a solution of compound BA-4-c (35.60 mg, 16.12 pmol, 1 eq) in DMF (1.5 mL) was added Et2NH (6 mg, 80.60 pmol, 5 eq) at 0 °C and stirred at 25 °C for 1 h, LCMS showed the starting material was consumed, oil pump concentrated directly to the next step. The theoretical mass of BA-4-d was 32 mg.

[0606] LCMS (ESI): m / z, 993 [1 / 2 M+H] + Rt = 1.739 min.

[0607] Step 5: Preparation of compound BA-4

[0608] To a solution of compound BA-4-d (32 mg, 16.11 μmol, 1 eq) and LY-22CD-3-c (the synthesis of this compound please refer to Preparation Example 41) (8.34 mg, 17.72 μmol, 1.1 eq) in DMF (1.5 mL) was added DIPEA (5.38 μL, 32.22 μmol, 2 eq) at 0 °C. After addition, the reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: YMC-Triart Prep C18 150×30mm×10μm. Mobile phase: water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 30% to 60% elution) and lyophilized to give compound BA-4 (5.74 mg, yield 14.50%, purity 95.29%) as a white solid.

[0609] LCMS (ESI): m / z, 1193 [1 / 2 M+Na] + Rt = 1.871 min.

[0610] 1H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 9.09 (s, 1H), 8.86 (d, J = 8.6 Hz, 1H), 8.38 (d, J = 8.6 Hz, 1H), 8.22 (d, J = 6.8 Hz, 1H), 8.19 - 8.11 (m, 2H), 8.03 (d, J = 7.6 Hz, 1H), 7.81 - 7.72 (m, 2H), 7.57 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.4 Hz, 2H), 7.09 (s, 1H), 6.20 (d, J = 3.0 Hz, 1H), 5.94 (d, J = 3.0 Hz, 1H), 5.42 (s, 2H), 4.94 (s, 2H), 4.87 (s, 1H), 4.81 (s, 1H), 4.75 (s, 1H), 4.54 (s, 5H), 4.42 - 4.26 (m, 5H), 4.19 (d, J = 7.6 Hz, 3H), 4.05 (s, 1H), 4.00 (s, 3H), 3.86 - 3.45 (m, 63H), 3.19 (s, 3H), 2.96 (d, J = 17.0 Hz, 5H), 2.75 (d, J = 12.8 Hz, 2H), 2.46 - 2.36 (m, 4H), 2.28 (t, J = 6.6 Hz, 2H), 2.13 (s, 2H), 1.95 (s, 4H), 1.84 (d, J = 11.8 Hz, 2H), 1.70 (d, J = 16.4 Hz, 5H), 1.62 (s, 4H), 1.45 (s, 5H), 1.30 (d, J = 7.2 Hz, 6H), 1.24 (s, 3H), 1.04 (d, J = 6.4 Hz, 3H), 0.86 (d, J = 6.8 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H).

[0611] Example 5: Preparation of compound BA-5

[0612] Step 1: Preparation of compound BA-5-a

[0613] To a solution of compound eribulin mesylate (10 mg, 0.012 mmol, 1 eq) in DMF (1 mL) was added Fmoc-Val-Ala-PAB-PNP (9.89 mg, 0.015 mmol, 1.2 eq), DIPEA (0.006 mL, 0.036 mmol, 3 eq) and HOBt (1.64 mg, 0.012 mmol, 1 eq) at 20 °C and stirred at room temperature for 1 h, LCMS showed the starting material was consumed completely. The reaction solution was purified by reverse column C18 (H2O (0.1% TFA) ACN = 17%:83%) to give the product BA-5-a (11 mg, yield 71.47%) as a white solid.

[0614] LCMS (ESI): m / z, 1293.2 [(M+Na)] + .

[0615] Step 2: Preparation of compound BA-5-b

[0616] To a solution of compound BA-5-a (11 mg, 0.009 mmol, 1 eq) in DMF (1 mL) was added diethylamine (0.004 mL, 0.043 mmol, 5 eq) at 0 °C and stirred at 20 °C for 1 h, LCMS showed the starting material was consumed completely. The reaction solution was purified by silica gel column chromatography (C18, H2O (0.1% TFA):ACN = 40%:60%) to give the product BA-5-b (7 mg, yield 77.09%) as a white solid. LCMS (ESI): m / z, 1048.7 [(M-1)] + .

[0617] Step 3: Preparation of compound BA-5

[0618] To a solution of compound BA-5-b (7 mg, 0.007 mmol, 1 eq) in DMF (1 mL) was added DIEA (0.002 mL, 0.013 mmol, 2 eq), 1-maleimido-3-oxo-7,10,13,16,19,22,25,28- octaoxa-4-aza (4.74 mg, 0.008 mmol, 1.2 eq) and HATU (3.04 mg, 0.008 mmol, 1.2 eq) at 0 °C and stirred at 25 °C for 1 h, LCMS showed the starting material was consumed completely. The reaction solution was purified by silica gel column chromatography (C18, H2O (0.1% TFA):MeCN = 34%:66%) to give the product BA-5 (5.2 mg, yield 48.01%) as a white solid.

[0619] LCMS (ESI): m / z, 1645.4 [(M+Na)]+ .

[0620] 1 H NMR (400 MHz, DMSO-d6) δ 9.92 (s, 1H), 8.18 (d, J = 6.8 Hz, 1H), 8.01 (s, 1H), 7.88 (d, J = 8.8 Hz, 1H), 7.58 (d, J = 8.4 Hz, 2H), 7.28 (d, J = 8.8 Hz, 2H), 7.09 (s, 1H), 7.00 (s, 2H), 5.03 (d, J = 22.8 Hz, 2H), 4.94 (s, 2H), 4.84 (s, 1H), 4.76 (s, 1H), 4.64 (s, 1H), 4.56 (t, J = 4.0 Hz, 1H), 4.44 - 4.36 (m, 1H), 4.27 (d, J = 10.8 Hz, 1H), 4.24 - 4.15 (m, 2H), 4.11 (s, 2H), 4.03 (s, 1H), 3.84 - 3.67 (m, 4H), 3.59 (dd, J = 12.0, 6.8 Hz, 5H), 3.49 (dd, J = 6.8, 4.4 Hz, 36H), 3.25 (s, 3H), 3.15 (dd, J = 11.6, 6.0 Hz, 3H), 2.97 (s, 2H), 2.84 (d, J = 10.4 Hz, 1H), 2.81 - 2.64 (m, 3H), 2.36 - 2.17 (m, 8H), 2.05 - 1.84 (m, 7H), 1.69 (dd, J = 25.2, 12.0 Hz, 5H), 1.51 (s, 3H), 1.31 (d, J = 7.2 Hz, 5H), 1.12 - 0.93 (m, 5H), 0.86 (dd, J = 16.0, 6.8 Hz, 7H).

[0621] Example 6: Preparation of compound BA-6

[0622] Step 1: Preparation of compound BA-6-a

[0623] To a solution of compound eribulin mesylate (15 mg, 0.018 mmol, 1 eq) in DMF (2 mL) was added Fmoc-L-valyl-L-alanine (8.94 mg, 0.022 mmol, 1.2 eq), DIPEA (0.009 mL, 0.054 mmol, 3 eq) and HATU (8.29 mg, 0.022 mmol, 1.2 eq) at 0 °C, and stirred at room temperature for 1 hour, LCMS showed the starting material was completely reacted. The reaction was prepared by silica gel column chromatography (C18) (0.1% TFA) H2O:ACN = 13%:87%), to give the product BA-6-a (16 mg, yield 78.51%) as a white solid.

[0624] LCMS (ESI): m / z, 1121.7 [(M-1)] + .

[0625] Step 2: Preparation of compound BA-6-b

[0626] To a solution of compound BA-6-a (16 mg, 0.014 mmol, 1 eq) in DMF (1 mL) was added diethylamine (5 mg, 0.07 mmol, 5 eq) at 0 °C and stirred at 20 °C for 1 h, LCMS showed the starting material was consumed. The reaction was concentrated by reduced pressure to get the crude (12.83 mg, crude), which was used in the next step directly without any purification.

[0627] LCMS (ESI): m / z, 900.2 [(M)] + .

[0628] Step 3: Preparation of compound BA-6

[0629] To a solution of compound BA-6-b (12.83 mg, 0.014 mmol, 1 eq) in DMF (2 mL) was added DIEA (0.007 mL, 0.043 mmol, 3 eq), LY-22CD-3-d (the synthesis of this compound please refer to Preparation Example 41) (12.49 mg, 0.016 mmol, 1.1 eq) and HATU (7.05 mg, 0.019 mmol, 1.3 eq) at 0 °C and stirred at 25 °C for 1 h, LCMS showed the starting material was consumed. The reaction was purified by high performance liquid chromatography preparation (Model JB-C235-04, column Bonnasil-BS C18, 20 x 250 mm x 8 μm, mobile phase water (0.1% TFA) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to get the white solid product BA-6 (7.44 mg, yield 31.09%).

[0630] LCMS (ESI): m / z, 1678.4 [(M+1)] + .

[0631] 1H NMR (400 MHz, DMSO-d6) δ 9.06 (t, J = 5.6 Hz, 1H), 8.87 (d, J = 8.4 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 7.96 (d, J = 7.3 Hz, 1H), 7.92 - 7.83 (m, 2H), 7.71 (t, J = 5.5 Hz, 1H), 5.03 (d, J = 21.5 Hz, 2H), 4.84 (s, 1H), 4.76 (s, 1H), 4.64 (s, 1H), 4.56 (t, J = 4.1 Hz, 1H), 4.30 - 4.24 (m, 2H), 4.20 - 4.14 (m, 2H), 4.11 (s, 3H), 4.02 (s, 1H), 3.84 - 3.71 (m, 3H), 3.65 (d, J = 3.5 Hz, 4H), 3.63 (s, 1H), 3.62 - 3.58 (m, 3H), 3.56 (dd, J = 10.2, 4.2 Hz, 5H), 3.51 - 3.48 (m, 26H), 3.25 (s, 4H), 3.19 (dd, J = 11.6, 5.8 Hz, 3H), 3.15 - 3.07 (m, 2H), 3.00 (d, J = 13.4 Hz, 2H), 2.88 - 2.66 (m, 5H), 2.37 - 2.18 (m, 10H), 2.02 - 1.89 (m, 7H), 1.76 - 1.45 (m, 9H), 1.24 (dd, J = 34.3, 17.4 Hz, 8H), 1.01 (dd, J = 23.2, 9.3 Hz, 4H), 0.83 (dd, J = 11.2, 6.8 Hz, 7H).

[0632] Example 7: Preparation of compound LA-1

[0633] Step 1: Preparation of compound HM-1118C_3

[0634] HM-1118C_1 (8 g, 20.2 mmol), HM-1118C_2 (3.66 g, 20.2 mmol) were added into a 250 mL three-necked flask, DMF (80 mL) was added, protected by nitrogen, DIPEA (7.8 g, 60.6 mmol) was added dropwise, stirred for 5 minutes after adding, HATU (9.2 g, 24.2 mmol) solution in DMF (30 mL) was added dropwise, stirred at room temperature for 1 hour, LCMS showed that the reaction was completed. The reaction solution was slowly poured into water (800 mL), stirred for 30 minutes, filtered, the solid was dissolved in DCM (200 mL), separated, washed once with water, the organic phase was dried over anhydrous sodium sulfate, rotary evaporated to give 9.5 g of yellow oil, yield 90%.

[0635] LCMS (ESI): m / z, 524.2 [M+H] + , 546.2 [M+Na] + .

[0636] 1 H NMR (400 MHz, DMSO) δ 8.32 (d, J = 6.7 Hz, 1H), 7.88 (d, J = 7.5 Hz, 2H), 7.70 (d, J = 7.3 Hz, 3H), 7.51 (t, J = 6.0 Hz, 1H), 7.41 (t, J = 7.4 Hz, 2H), 7.32 (t, J = 7.4 Hz, 2H), 4.19 (m, 5H), 3.66 (d, J = 5.1 Hz, 2H), 1.95 (m, 1H), 1.37 (s, 9H), 1.23 (d, J = 7.2 Hz, 3H), 0.85 (dd, J = 18.7, 6.8 Hz, 6H).

[0637] Step 2: Preparation of compound HM-1118C_4

[0638] HM-1118C_3 (6 g, 11.47 mmol) was dissolved in DCM (30 mL) under nitrogen protection, TFA (20 mL) was added dropwise, and the reaction was carried out at room temperature for 2 hours. LCMS showed that the reaction was complete. The reaction solution was concentrated to a small volume, saturated sodium bicarbonate was added to pH = 7-8, DCM (100 mL) was added to extract the impurities, 1N HC1 was added to adjust the pH to 1-2, the product was extracted with EA (200 mL x 2), the organic phase was washed with water (100 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and rotary evaporated to give 4.8 g of white solid with a yield of 89%.

[0639] LCMS (ESI): m / z, 468.2 [M+H] + , 490.2 [M+Na] + .

[0640] 1 H NMR (400 MHz, DMSO) δ 7.88 (d, J = 7.5 Hz, 2H), 7.70 (d, J = 7.4 Hz, 2H), 7.41 (t, J = 7.4 Hz, 2H), 7.32 (t, J = 7.3 Hz, 2H), 4.21 (m, 5H), 3.65 (s, 2H), 1.92 (m, 1H), 1.26 (d, J = 7.3 Hz, 3H), 0.84 (dd, J = 20.8, 6.7 Hz, 6H).

[0641] Step 3: Preparation of compound HM-1118C_5

[0642] HM-1118C_4 (2.5 g, 5.35 mmol), (4-aminophenyl)methanol (645 mg, 5.24 mmol) were added to a reaction bottle, dissolved in DMF (25 mL), protected by nitrogen, 2,6-dimethylpyridine (1.72 g, 16 mmol) was added, stirred for 5 minutes after addition, dropwise addition of HATU (2.44 g, 6.4 mmol) in DMF (8 mL), reaction for 1 hour at room temperature. LCMS showed that the reaction was complete. The reaction was added to water (200 mL), the product was extracted with EA (200 mL x 2), the organic phase was washed with dilute hydrochloric acid, the organic phase was washed with sodium bicarbonate, the organic phase was dried over anhydrous sodium sulfate, and rotary evaporation was performed to obtain 2.4 g of nearly white solid with a yield of 80%.

[0643] LCMS (ESI): m / z, 595.2 [M+Na] + .

[0644] 1 H NMR (400 MHz, DMSO) δ 7.90 (d, J = 7.5 Hz, 2H), 7.70 (d, J = 7.5 Hz, 2H), 7.55 (d, J = 8.3 Hz, 2H), 7.43 (t, J = 7.4 Hz, 2H), 7.33 (t, J = 7.4 Hz, 2H), 7.24 (d, J = 8.3 Hz, 2H), 4.48-4.35 (m, 3H), 4.25 (dd, J = 21.4, 6.6 Hz, 4H), 3.70 (d, J = 2.4 Hz, 2H), 2.00-1.95 (m, 1H), 1.33 (d, J = 7.1 Hz, 3H), 0.87 (dd, J = 19.4, 6.8 Hz, 6H).

[0645] Step 4: Preparation of compound HM-1118C_6

[0646] HM-1118C_5 (1 g, 1.74 mmol), HM-1118C_1A (0.8 g, 2.62 mmol) were dissolved in DMF (10 mL), protected by nitrogen, dropwise addition of DIPEA (563 mg, 4.37 mmol), reaction for 2 hours at room temperature, LCMS showed that the reaction was complete. The reaction was added to water (50 mL), the product was extracted with EA (100 mL x 2), the EA phase was dried over anhydrous sodium sulfate and concentrated, the sample was mixed with silica gel, and column chromatography was performed using a PE:EA system, and the product was eluted with pure EA, rotary evaporation was performed to obtain 1 g of nearly white solid with a yield of 78%.

[0647] LCMS (ESI): m / z, 760.2 [M+Na] + .

[0648] 1H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 8.35 - 8.23 (m, 3H), 7.87 (m, 3H), 7.62 (m, 7H), 7.41 (t, J = 6.8 Hz, 4H), 7.32 (t, J = 7.0 Hz, 2H), 5.23 (s, 2H), 4.45 - 4.33 (m, 1H), 4.22 (m, 4H), 3.70 (s, 2H), 1.99 (m, 1H), 1.33 (m, 3H), 0.86 (dd, J = 19.9, 6.8 Hz, 6H).

[0649] Step 5: Preparation of compound HM-1118C_7

[0650] HM-1118C_6 (100 mg, 0.135 mmol), HY-308_ABC10 (110 mg, 0.133 mmol) were added to the reaction bottle, protected by nitrogen, dissolved in DMF (2 mL), HOBT (18.3 mg, 0.135 mmol) was added, DIPEA (70 mg, 0.542 mmol) was added, and the reaction was carried out at room temperature for 1 hour. LC-MS showed that the reaction was complete. The reaction solution was poured into ice water, EA (100 mL) was added, the product was extracted, the organic phase was washed with water 50 mL x 3, the organic phase was dried over anhydrous sodium sulfate, rotary evaporation, and the product was obtained by using PE:EA = 10:1 to wash and filter, 150 mg of white solid was obtained, the yield was 83%.

[0651] LCMS (ESI): m / z, 1350.4 [M+Na] + .

[0652] 1H NMR (400 MHz, DMSO) δ 7.87 (d, J = 7.7 Hz, 2H), 7.67 (d, 2H), 7.56 (d, J = 8.3 Hz, 2H), 7.40 (t, J = 7.4 Hz, 2H), 7.32 (t, J = 6.0 Hz, 2H), 7.25 (d, J = 8.5 Hz, 2H), 5.04 (s, 1H), 4.98 (s, 1H), 4.92 (d, 2H), 4.81 (s, 1H), 4.74 (s, 1H), 4.63 (s, 1H), 4.54 (t, 1H), 4.36 (d, J = 7.5 Hz, 1H), 4.24 (m, 3H), 4.20 (m, 3H), 4.09 (s, 3H), 4.01 (m, 1H), 3.77 (m, 2H), 3.68 (d, 2H), 3.23 (s, 4H), 2.95 (s, 2H), 2.83 (m, 4H), 2.30 - 2.09 (m, 6H), 2.01 - 1.88 (m, 7H), 1.72 (m, 6H), 1.52 (s, 3H), 1.33 - 1.21 (m, 9H), 1.01 (m, 4H), 0.84 (m, 7H).

[0653] Step 6: Preparation of compound HM-1118C_8

[0654] HM-1118C_7 (170 mg, 0.128 mmol) was dissolved in DCM (3 mL), piperidine (0.3 mL) was added dropwise, and the reaction was allowed to proceed at room temperature for 1 hour. LCMS showed that the reaction was complete. PE (150 mL) was added to the reaction solution, and an oil precipitated and adhered to the wall of the flask. After 10 minutes of precipitation, the supernatant was poured out, and the residual solvent was removed by oil pump. The residue was dissolved in solvent and purified by reverse-phase liquid chromatography (preparative chromatography manufacturer: Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10μm. The mobile phase was water (0.1% TFA)-acetonitrile, and the acetonitrile elution ratio was eluted from 10% to 40%). (TFA system), and 90 mg of white solid was obtained by freeze-drying, with a yield of 63%.

[0655] LCMS (ESI): m / z, 1106.5 [M+H] + .

[0656] 1H NMR (400 MHz, DMSO) δ 10.00 (s, 1H), 8.40 (t, J = 7.3 Hz, 2H), 7.94 (t, 3H), 7.56 (d, J = 8.6 Hz, 2H), 7.28 (d, J = 8.6 Hz, 2H), 7.07 (t, J = 5.5 Hz, 1H), 5.05 (s, 1H), 5.00 (s, 1H), 4.98-4.90 (m, 2H), 4.83 (s, 1H), 4.75 (s, 1H), 4.64 (m, 1H), 4.55 (t, J = 4.1 Hz, 1H), 4.43-4.32 (m, 2H), 4.26 (d, J = 10.4 Hz, 1H), 4.22-3.97 (m, 6H), 3.85-3.73 (m, 3H), 3.67 (m, 1H), 3.62 (m, 2H), 3.27-3.19 (m, 4H), 2.98 (t, J = 5.6 Hz, 2H), 2.84 (d, J = 9.7 Hz, 1H), 2.73 (m, 2H), 2.60-2.53 (m, 1H), 2.33-2.11 (m, 6H), 2.03-1.88 (m, 7H), 1.68 (m, 5H), 1.51 (m, 3H), 1.26 (m, 7H), 1.01 (m, 4H), 0.91-0.83 (m, 6H).

[0657] Step 7: Preparation of compound HM-1118B_2A

[0658] HM-1118C_2A (300 mg, 0.97 mmol), HM-1118A_7 (the synthesis of this compound please refer to the example of LA-3) (789 mg, 2.43 mmol) were added into THF (5 mL), water (5 mL), NaHCO3 (409 mg, 48.7 mmol), and the reaction was carried out at room temperature for 1 hour. LCMS showed that the reaction was completed. The reaction solution was concentrated to a small volume, and then freeze-dried. Reverse phase liquid chromatography preparation (Oriendo, BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10μm. The mobile phase was water (0.1% TFA)-acetonitrile, and the elution ratio of acetonitrile was from 20% to 40% elution) (TFA system) to obtain 65 mg of white solid, with a yield of 13%.

[0659] LCMS (ESI): m / z, 518.2 [M+H] + .

[0660] 1H NMR (400 MHz, DMSO) δ 12.42 (s, 1H), 7.98 (d, J = 7.7 Hz, 1H), 7.63 (t, J = 5.9 Hz, 1H), 7.00 (s, 2H), 4.11 (m, 1H), 3.97 (d, J = 3.7 Hz, 1H), 3.90 (d, J = 1.9 Hz, 1H), 3.60 - 3.54 (m, 2H), 3.46 (m, 3H), 3.39 - 3.35 (m, 6H), 3.10 - 3.02 (m, 2H), 2.09 (t, J = 7.2 Hz, 2H), 1.53 (m, 8H), 1.31 - 1.13 (m, 4H).

[0661] Step 8: Preparation of compound LA-1

[0662] HM-1118C_8 (72 mg, 0.065 mmol), HM-1118B_2A (40 mg, 0.065 mmol) were dissolved in DMF (2 mL), DIPEA (25 mg, 0.19 mmol) was added, dropwise added HATU (25 mg, 0.065 mmol) solution in DMF (0.1 mL), after adding, stirred at room temperature for 30 minutes, LCMS showed that the reaction was completed. The reaction solution was directly prepared by reversed-phase liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10μm. The mobile phase was water (0.1% TFA)-acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%), and then the preparation liquid was prepared again by a neutral column (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10μm. The mobile phase was water (10mM NH4HCO3)-acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) to remove TFA, and 46 mg of white solid was obtained after freeze-drying, with a yield of 44%.

[0663] LCMS (ESI): m / z, 1605.7 [M+H] +

[0664] 1H NMR (400 MHz, DMSO) δ 9.89 (s, 1H), 8.24-8.11 (m, 2H), 7.92 (d, J = 7.5 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.63-7.54 (m, 3H), 7.27 (d, J = 8.4 Hz, 2H), 7.08 (t, 1H), 7.00 (s, 2H), 5.35 (d, J = 5.0 Hz, 1H), 5.05 (s, 1H), 5.00 (s, 1H), 4.93 (s, 2H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.55 (m, 3H), 4.47 (d, J = 5.2 Hz, 1H), 4.38 (m, 2H), 4.31 (t, 1H), 4.24 (d, 1H), 4.19 (m, 3H), 4.10 (s, 3H), 4.03-3.96 (m, 2H), 3.91 (m, 1H), 3.77 (m, 5H), 3.49 (m, 6H), 3.39 (s, 2H), 3.24 (s, 1H), 3.05 (m, 2H), 2.97 (m, 2H), 2.84 (d, J = 10.2 Hz, 1H), 2.74 (m, 2H), 2.31-2.07 (m, 8H), 1.95 (m, 7H), 1.68 (m, 6H), 1.54-1.44 (m, 8H), 1.31 (m, 9H), 1.18 (m, 4H), 1.00 (m, 5H), 0.84 (dd, J = 21.0, 6.7 Hz, 7H).

[0665] Example 8: Preparation of compound LA-2

[0666] Step 1: Preparation of compound HM-1118B_1

[0667] The starting material HM-297D_9 (1 g, 3.24 mmol, 1 eq) was dissolved in DMF (10 mL, 10V), HM-1118A_7 (1.58 g, 4.87 mmol, 1.5 eq), DIPEA (1.26 g, 9.73 mmol, 3 eq) were added, after addition, the reaction was reacted at 10-20 °C for 16 hours. LCMS showed that part of the starting material was left, the reaction liquid was separated by liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10um. The mobile phase was water (0.1% TFA)-acetonitrile, and the elution ratio of acetonitrile was from 30% to 60% elution) (TFA system) to obtain 320 mg of product, white solid, yield 19%.

[0668] 1H NMR (400 MHz, D20) δ 6.76 (s, 2H), 4.32 - 4.17 (m, 2H), 4.07 - 3.95 (m, 1H), 3.78 - 3.63 (m, 3H), 3.58 (dd, J = 11.6, 6.0 Hz, 1H), 3.43 (t, J = 6.9 Hz, 2H), 3.19 (t, J = 6.9 Hz, 2H), 2.22 (t, J = 7.2 Hz, 2H), 1.87 - 1.75 (m, 1H), 1.68 (ddd, J = 14.2, 11.9, 7.3 Hz, 1H), 1.61 - 1.47 (m, 6H), 1.43 - 1.26 (m, 2H), 1.26 - 1.14 (m, 2H). LCMS 518.2 [M+l] +

[0669] Step 2: Preparation of compound HM-297D_5

[0670] The starting material HM-297D_4 (5.0 g, 8.3 mmol) was suspended in super dry DCM (50 mL), added DEA (diethylamine, 25 mL), the starting material gradually dissolved, reacted at room temperature (10-20 °C) for 2 hours. LCMS showed that the starting material was completely converted, the reaction liquid was spin dried (30-40 °C), the crude product was dissolved in dichloromethane (50 mL) and spin dried twice, then recrystallized with isopropyl alcohol (20V) to obtain HM-297D_5 (3.0 g, 95% yield) white powdery solid.

[0671] 1 H NMR (400 MHz, DMSO) δ 10.02 (s, 1H), 8.11 (d, J = 6.8 Hz, 1H), 7.54 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 8.5 Hz, 2H), 5.96 (t, J = 5.7 Hz, 1H), 5.40 (s, 2H), 5.09 (t, J = 5.7 Hz, 1H), 4.45 (dd, J = 16.0, 5.4 Hz, 3H), 3.16 - 2.80 (m, 3H), 2.07 - 1.85 (m, 1H), 1.79 - 1.50 (m, 4H), 1.50 - 1.25 (m, 2H), 0.89 (d, J = 6.9 Hz, 3H), 0.78 (d, J = 6.8 Hz, 3H). LCMS 380.3 [M+l] + .

[0672] Step 3: Preparation of compound HM-1118B_4

[0673] The starting material HM-297D_5 (1.5 g, 3.96 mmol, 1 eq) was dissolved in super dry DMF (30 mL), HM-1118B_3 (1.59 g, 4.0 mmol, 1.02 eq) was added, DIPEA (765.3 mg, 5.9 mmol, 1.5 eq) was added dropwise to the reaction solution, the reaction solution was clear, and the reaction was carried out in an ice bath (0-5 °C) for 1 h. LCMS showed that the starting material was completely converted, at this time the reaction solution precipitated solid, EA (150 mL) was added dropwise to the reaction solution, after the addition was completed, it was continuously stirred for 30 min, then filtered, and the solid was collected to obtain the product HM-1118B_4 (2.5 g, 96.1% yield) in white powder form.

[0674] 1 H NMR (400 MHz, DMSO) δ 9.88 (s, 1H), 8.20 (d, J = 7.3 Hz, 1H), 7.88 (dd, J = 21.5, 8.0 Hz, 3H), 7.71 (d, J = 6.9 Hz, 2H), 7.66 - 7.51 (m, 3H), 7.43 (t, J = 7.4 Hz, 2H), 7.34 (t, J = 7.4 Hz, 2H), 7.24 (d, J = 8.3 Hz, 2H), 6.01 (s, 1H), 5.44 (s, 2H), 5.14 (t, J = 5.6 Hz, 1H), 4.51 - 4.34 (m, 3H), 4.25 (dt, J = 20.7, 7.0 Hz, 4H), 3.72 (d, J = 6.0 Hz, 2H), 3.09 - 2.93 (m, 2H), 2.11 - 1.92 (m, 1H), 1.80 - 1.56 (m, 2H), 1.54 - 1.31 (m, 2H), 0.86 (dd, J = 17.5, 6.7 Hz, 6H). LCMS 659.4 [M+l] + .

[0675] Step 4: Preparation of compound HM-1118B_5

[0676] The starting material HM-1118B_4 (450 mg, 0.68 mmol, 1 eq) was suspended in DMAC (N,N-dimethylacetamide, 13.5 mL, 30 V), NPC (517.2 mg, 1.7 mmol, 2.5 eq) was added, DIPEA (132 mg, 1.0 mmol, 1.5 eq) was added dropwise to the reaction solution, the reaction solution was turbid, and the reaction was carried out at (20-30 °C) for 48 h. After the reaction was completed, the reaction solution was added dropwise to MTBE (methyl tert-butyl ether, 67.5 mL), flocculent precipitated, filtered, and the product was collected.

[0677] 1H NMR (400 MHz, DMSO) δ 10.05 (s, 1H), 8.29 (dd, J = 30.8, 8.2 Hz, 3H), 7.88 (dd, J = 14.6, 8.1 Hz, 3H), 7.77 - 7.50 (m, 7H), 7.39 (ddd, J = 35.2, 10.3, 4.9 Hz, 6H), 6.02 (d, J = 5.1 Hz, 1H), 5.46 (s, 2H), 5.26 (s, 2H), 4.42 (dd, J = 13.1, 7.7 Hz, 1H), 4.26 (dq, J = 13.7, 6.9 Hz, 4H), 3.73 (d, J = 5.9 Hz, 2H), 3.14 - 2.97 (m, 2H), 2.07 - 1.99 (m, 1H), 1.84 - 1.57 (m, 2H), 1.44 (dd, J = 15.8, 8.7 Hz, 2H), 0.88 (dd, J = 18.0, 6.7 Hz, 6H). LCMS 824.3 [M+l] + .

[0678] Step 5: Preparation of compound HM-1118B_6

[0679] The starting material eribulin mesylate (30 mg, 36.33 μmol, 1 eq) was dissolved in DMAC (N,N-dimethylacetamide, 0.3 mL, 10V), DIPEA (4.69 mg, 36.33 μmol, 1 eq, 0.1 mL DMAC dilution) was added, after the reaction was stirred for 10 minutes, HM-1118B_5 (41.89 mg, 51 μmol, 1.4 eq), HOBt (2.43 mg, 18 μmol, 0.1 mL DMAC dilution) and pyridine (5.74 mg, 72.7 μmol, 2 eq, 0.1 mL DMAC dilution) were added in turn, after the addition was completed, the reaction was reacted at 5-10 °C for 16 hours. After LCMS showed that the reaction was completed, the reaction liquid was added dropwise into MTBE (5 mL), the flocculent precipitated, was filtered, and the product was collected.

[0680] 1H NMR (400 MHz, DMSO) δ 9.95 (s, 1H), 8.37 (s, 1H), 8.19 (d, J = 7.2 Hz, 1H), 7.85 (dd, J = 26.8, 8.0 Hz, 3H), 7.73 - 7.66 (m, 2H), 7.64 - 7.49 (m, 3H), 7.41 (t, J = 7.3 Hz, 2H), 7.28 (dt, J = 24.3, 11.6 Hz, 4H), 7.07 (t, J = 5.6 Hz, 1H), 5.99 (t, J = 5.4 Hz, 1H), 5.41 (s, 2H), 5.10 - 4.87 (m, 4H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.55 (t, J = 4.2 Hz, 2H), 4.38 (dd, J = 13.3, 7.8 Hz, 1H), 4.31 - 4.13 (m, 6H), 4.02 (s, 4H), 3.86 - 3.45 (m, 10H), 3.25 (d, J = 7.5 Hz, 3H), 3.13 (d, J = 7.0 Hz, 1H), 3.02 (d, J = 6.8 Hz, 1H), 2.84 (d, J = 9.7 Hz, 1H), 2.73 - 2.65 (m, 1H), 2.62 - 2.55 (m, 1H), 2.28 - 2.09 (m, 4H), 2.00 (d, J = 7.4 Hz, 2H), 1.92 (s, 3H), 1.80 - 1.61 (m, 6H), 1.59 - 1.41 (m, 4H), 1.27 (t, J = 12.5 Hz, 13H), 1.00 (dd, J = 21.3, 9.0 Hz, 4H), 0.84 (dd, J = 17.8, 6.7 Hz, 6H). LCMS 1414.6 [M+l] + .

[0681] Step 6: Preparation of compound HM-1118B_7

[0682] The starting material HM-1118B_6 (45 mg, 31.8 pmol, 1 eq) was dissolved in DMAC / DEA (N,N-dimethylacetamide / triethylamine, 5:1, 0.67 mL, 15 V) and the reaction was allowed to react at 5-10 °C for 2 hours. After LCMS showed the reaction was completed, the reaction was separated by liquid chromatography preparation (preparation chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10pm. The mobile phase was water (0.1% TFA)-acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%) to obtain 14 mg of product (TFA system).

[0683] 1H NMR (400 MHz, DMSO) δ 10.12 (s, 1H), 8.43 (dd, J = 31.1, 8.2 Hz, 2H), 8.05 (s, 3H), 7.63 (d, J = 8.5 Hz, 2H), 7.34 (d, J = 8.5 Hz, 2H), 7.13 (t, J = 5.7 Hz, 1H), 6.11 (s, 1H), 5.52 (s, 2H), 5.05 (dd, J = 27.2, 17.9 Hz, 4H), 4.89 (s, 1H), 4.81 (s, 1H), 4.70 (s, 1H), 4.61 (d, J = 4.2 Hz, 1H), 4.51 - 4.38 (m, 2H), 4.32 (d, J = 10.4 Hz, 1H), 4.28 - 4.01 (m, 5H), 3.93 - 3.57 (m, 9H), 3.31 (d, J = 4.7 Hz, 3H), 3.12 - 2.96 (m, 4H), 2.93 - 2.63 (m, 4H), 2.48 - 2.14 (m, 6H), 2.13 - 1.90 (m, 7H), 1.86 - 1.31 (m, 15H), 1.30 - 1.18 (m, 2H), 1.07 (dd, J = 23.1, 9.0 Hz, 4H), 0.93 (dd, J = 18.2, 6.8 Hz, 6H). LCMS 1192.5 [M+l] + ,597 [1 / 2 M+l].

[0684] Step 7: Preparation of compound LA-2

[0685] The starting material HM-1118B_7 (40 mg, 33.56 pmol, 1 eq) was dissolved in DMF (2 mL), HM-1118B_1 (20.82 mg, 40.27 pmol, 1.2 eq) was added, cooled to 0 °C in an ice bath, HATU (15.3 mg, 40.27 pmol, 1.2 eq) was added, DIPEA (13 mg, 100.68 pmol, 3 eq), after reaction at 0-5 °C for two hours, liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10pm. The mobile phase was water (0.1% TFA)-acetonitrile, and the elution ratio of acetonitrile was from 20% to 60% elution) was carried out, and the product (24 mg, yield 42%) was separated as a white solid.

[0686] 1H NMR (400 MHz, DMSO) δ 9.96 (s, 1H), 8.19 (s, 2H), 7.95 (d, J = 7.3 Hz, 1H), 7.74 (d, J = 8.7 Hz, 1H), 7.62 (dd, J = 18.0, 7.1 Hz, 3H), 7.29 (d, J = 8.5 Hz, 2H), 7.11 (d, J = 8.1 Hz, 1H), 7.01 (d, J = 9.6 Hz, 2H), 6.01 (s, 1H), 5.47 - 5.34 (m, 3H), 4.99 (d, J = 26.0 Hz, 4H), 4.81 (d, J = 31.9 Hz, 2H), 4.51 (dd, J = 50.5, 22.6 Hz, 7H), 4.28 - 4.12 (m, 7H), 4.02 (d, J = 17.5 Hz, 2H), 3.80 (dd, J = 32.0, 7.9 Hz, 5H), 3.59 - 3.50 (m, 5H), 3.27 (s, 4H), 3.15 - 2.61 (m, 12H), 2.34 - 1.93 (m, 14H), 1.82 - 1.62 (m, 8H), 1.61 - 1.19 (m, 25H), 1.03 (dd, J = 22.6, 8.9 Hz, 4H), 0.85 (dd, J = 18.0, 6.7 Hz, 6H).

[0687] LCMS 1691.7 [M+l] + , 846.6 [1 / 2M+l] + .

[0688] Example 9: Preparation of compound LA-3

[0689] Step 1: Preparation of compound HM-1118A_3

[0690] The starting material TEA (triethylamine, 753 mg, 7.4 mmol), HM-1118A_2 (659 mg, 2.4 mmol) was dissolved in super dry DMF (15 mL), HM-1118A_1 (1.0 g, 2.4 mmol) was added, and the reaction was carried out at room temperature (10-20 °C) for 17 hours. LCMS showed that the starting material was completely converted, and the residue was purified by silica gel column chromatography (C18) (ISCO, R-330g SepaFlash Silica Flash Column, Eluent of 5-75% water / CH3CN @ 100 mL / min) to give 1.2 g, yield 87% after lyophilization.

[0691] LCMS (ESI): m / z, 553.2 [M+H] + , 575.2 [M+Na] +1H NMR (400 MHz, DMSO) δ 7.71 - 7.65 (m, 1H), 7.62 (d, J = 7.0 Hz, 1H), 7.54 - 7.44 (m, 3H), 7.41 - 7.28 (m, 3H), 5.04 (d, J = 14.0 Hz, 1H), 3.65 - 3.56 (m, 3H), 3.50 - 3.40 (m, 9H), 3.30 (t, J = 5.7 Hz, 2H), 3.13 - 3.03 (m, 2H), 2.62 - 2.54 (m, 4H), 2.40 (t, J = 6.4 Hz, 2H), 2.29 - 2.18 (m, 1H), 2.05 - 1.95 (m, 1H), 1.87 - 1.71 (m, 1H).

[0692] Step 2: Preparation of compound HM-1118A_4

[0693] To HM-1118A_3 (2.3 g, 4.16 mmol), HOSU (N-hydroxysuccinimide, 0.718 g, 6.24 mmol) was dissolved in DCM (40 mL), EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1.19 g, 6.24 mmol) was added under stirring, and the reaction was carried out at 10-15 °C for 16 h. LCMS showed that the reaction was completed, water (20 mL) was added to the reaction solution, the aqueous phase was separated, and the organic phase was extracted with DCM (40 mL), and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 2.1 g of a light yellow foam, with a yield of 77%.

[0694] LCMS (ESI): m / z, 650.2 [M+H] + , 672.2 [M+Na] + . 1 1H NMR (400 MHz, DMSO) δ 7.81 (t, J = 5.5 Hz, 1H), 7.77 - 7.72 (m, 1H), 7.70 - 7.66 (m, 1H), 7.59 - 7.49 (m, 3H), 7.48 - 7.35 (m, 3H), 5.09 (d, J = 14.0 Hz, 1H), 3.77 (t, J = 6.0 Hz, 2H), 3.68 (dd, J = 12.5, 5.4 Hz, 1H), 3.64 - 3.43 (m, 14H), 3.35 (d, J = 6.2 Hz, 2H), 3.19 - 3.10 (m, 2H), 2.97 (t, J = 6.0 Hz, 2H), 2.69 - 2.60 (m, 1H), 2.35 - 2.25 (m, 1H), 2.11 - 2.02 (m, 1H), 1.89 - 1.75 (m, 1H).

[0695] Step 3: Preparation of compound HM-1118A_6

[0696] The starting material HM-1118A_5 (5 g, 0.02 mol), compound of CAS: 90-80-2 (3.6 g, 0.02 mol) was dissolved in MeOH (80 mL), TEA (4.1 g, 0.04 mol) was added, and the reaction was carried out at 70 °C for 16 hours. LCMS showed that the starting material was completely converted. The reaction solution was concentrated, 50 mL of MTBE was added three times, and then concentrated to dryness. Finally, the crude product was obtained by slurry with petroleum ether, which was a white foam (the product was an acidic compound, and a salt was formed with TEA), and the yield was not calculated.

[0697] LCMS (ESI): m / z, 425.2 [M+H] + , 447.2 [M+Na] + . 1 H NMR (400 MHz, DMSO) δ 7.50 (brs, 1H), 5.61 (d, J = 4.0 Hz, 1H), 4.27 (s, 1H), 4.13 (s, 1H), 4.10-3.85 (m, 1H), 3.85-3.66 (m, 4H), 3.28-3.15 (m, 2H), 1.80-1.55 (m, 2H), 1.55-1.48 (m, 2H), 1.50-1.21 (m, 11H).

[0698] Step 4: Preparation of compound LA-1118A_7

[0699] The starting material HM-1118A_6 (7 g) was dissolved in DCM (35 mL), and TFA (35 mL) was added. The reaction was carried out at 10-15 °C for 16 hours. LCMS showed that the starting material was completely converted. The reaction solution was concentrated and added with DCM (50 mL x 3) and concentrated three times. After adding water, the freeze-drying was carried out, and the crude product was obtained as a yellow sticky substance, 8 g, and the yield was not calculated.

[0700] 1 H NMR (400 MHz, DMSO) δ 9.26 (s, 1H), 8.26 (d, J = 25.1 Hz, 3H), 7.69 (dd, J = 37.2, 31.4 Hz, 1H), 3.99 (d, J = 3.7 Hz, 1H), 3.94-3.84 (m, 2H), 3.58 (dd, J = 13.3, 3.7 Hz, 1H), 3.51-3.45 (m, 2H), 3.40-3.34 (m, 1H), 3.09 (qt, J = 15.7, 7.9 Hz, 2H), 1.84-1.67 (m, 2H), 1.60-1.27 (m, 4H), 1.18 (t, J = 7.3 Hz, 5H).

[0701] Step 5: Preparation of compound HM-1118A_8

[0702] To HM-1118A_7 (3 g, 9.6 mmol) was dissolved in DMF (40 mL), TEA (1.6 g, 16 mmol) was added to make the reaction system alkaline (pH greater than 9), then HM-1118A_4 (2.1 g, 3.2 mmol) was added in DMF (10 mL), and the reaction was carried out at 25-30 °C for 16 h. LCMS showed that the reaction was completed, and the residue was purified by silica gel column chromatography (C18) (ISCO, R-330g SepaFlash Silica Flash Column, Eluent of 5-75% water / CH3CN @ 100 mL / min), and freeze-dried to give 600 mg, with a yield of 22%.

[0703] LCMS (ESI): m / z, 859.3 [M+H] + , 882.3 [M+Na] + . 1 H NMR (400 MHz, DMSO) δ 7.75 (s, 2H), 7.71-7.55 (m, 3H), 7.55-7.42 (m, 3H), 7.40-7.23 (m, 3H), 5.03 (d, J = 14.0 Hz, 1H), 4.12-3.89 (m, 3H), 3.66-3.57 (m, 4H), 3.52-3.42 (m, 12H), 3.29-3.25 (m, 5H), 3.15-3.00 (m, 4H), 2.75-2.65 (m, 5H), 2.34-2.22 (m, 4H), 2.05-1.95 (m, 1H), 1.81-1.73 (m, 1H), 1.67-1.47 (m, 2H), 1.42-1.32 (m, 2H), 1.30-1.20 (m, 2H).

[0704] Step 6: Preparation of compound LA-3

[0705] To a solution of HM-1118A_8 (34 mg, 0.04 mmol), HM-1118B_7 (the synthesis of this compound please refer to the example of LA-2) (40 mg, 0.03 mmol) in DMF (2 mL) was cooled to 0-5 °C in ice bath, HATU (15 mg, 0.04 mmol), DIPEA (10.8 mg, 0.08 mmol) was added successively, and the mixture was kept at 0-5 °C for 1 h. LCMS showed the reaction was completed, the reaction solution was prepared by liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was YMC-Triart Prep C18 150*30mm*10μm. The mobile phase was water (0.1% TFA)-acetonitrile, and the acetonitrile elution ratio was eluted from 30% to 60%). 29 mg was obtained by freeze-drying, the yield was 42%.

[0706] LCMS (ESI): m / z, 1017.2 [M / 2+H] + . 1 H NMR (400 MHz, DMSO) δ 9.94 (s, 1H), 8.77 (d, J = 3.8 Hz, 1H), 8.54 (d, J = 7.7 Hz, 1H), 8.16 (s, 2H), 8.03 (d, J = 7.3 Hz, 1H), 7.78 - 7.66 (m, 3H), 7.60 (dd, J = 15.3, 7.8 Hz, 4H), 7.53 - 7.44 (m, 3H), 7.40 - 7.32 (m, 2H), 7.29 (t, J = 8.4 Hz, 3H), 7.08 (d, J = 4.9 Hz, 1H), 5.99 (s, 1H), 5.42 (s, 3H), 5.15 - 4.71 (m, 8H), 4.69 - 4.50 (m, 4H), 4.37 (d, J = 6.0 Hz, 2H), 4.29 - 3.98 (m, 11H), 3.96 - 3.64 (m, 9H), 3.61 - 3.54 (m, 6H), 3.48 (s, 11H), 3.16 - 2.80 (m, 13H), 2.40 (dd, J = 13.8, 7.1 Hz, 4H), 2.32 - 2.08 (m, 8H), 2.06 - 1.85 (m, 9H), 1.82 - 1.12 (m, 26H), 1.10 - 0.95 (m, 4H), 0.84 (dd, J = 17.4, 6.7 Hz, 6H).

[0707] Example 10: Preparation of compound LA-4

[0708] Step 1: Preparation of compound LA-4-b To a solution of compound LA-4-a (purchased from Haoyuan Pharmaceutical, 10.60 mg, 8.27 pmol, 1 eq) in DCM (5 mL) was added trifluoroacetic acid (0.5 mL) at 0 °C, after addition, the reaction was stirred at 0-5 °C for 5 h, LCMS showed the reaction was completed, the reaction solution was rotary evaporated to get yellow oil LA-4-b (47 mg, yield 90.00%).

[0709] LCMS (ESI): m / z, 602.2 [M+H] + Rt = 0.74 min.

[0710] Step 2: Preparation of compound LA-4-c

[0711] To a solution of compound LA-4-b (1 eq, 72 mg, 82.50 pmol, 82%) and LY-5a (1.6 eq, 73.07 mg, 132 pmol) in DMF (3 mL) was added DIPEA (7 eq, 74.64 mg, 577.50 pmol) and HATU (1.5 eq, 47.05 mg, 123.80 pmol) at 0 °C, after addition, the reaction was stirred at 25 °C for 12 h, LCMS showed the reaction was completed, the reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was YMC-Triart Prep C18 250 x 50 mm x 7 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution), and then lyophilized to get white solid compound LA-4-c (34.60 mg, yield 36.88%).

[0712] LCMS (ESI): m / z, 1045.6 [M+Na] + .

[0713] Step 3: Preparation of compound LA-4-d

[0714] To a solution of compound LA-4-c (30.60 mg, 26.91 μmol, 1 eq) and eribulin mesylate (23.90 mg, 26.91 μmol, 1 eq, purchased from Shandong Xingzhi) in DMF (2 mL) was added DIPEA (17.39 mg, 134.55 μmol, 5 eq) and HOBt (1.82 mg, 13.46 μmol, 0.5 eq) at 0 °C. After addition, the mixture was stirred at 25 °C for 12 h. LCMS showed the reaction was completed. The reaction solution was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: YMC-Triart Prep C18 250 x 50 mm x 7 μm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 48% to 78% elution) and lyophilized to give compound LA-4-d (1 mg, yield 25.34%) as a white solid.

[0715] LCMS (ESI): m / z, 1636.0 [M+Na] + Rt = 2.82 min.

[0716] Step 4: Preparation of compound LA-4-e

[0717] To a solution of compound LA-4-d (11 mg, 6.82 μmol, 1 eq) in DCM (5 mL) was added trifluoroacetic acid (0.5 mL) at 0 °C. After addition, the mixture was stirred at 0-10 °C for 3 h. LCMS showed the reaction was completed. The reaction solution was rotary evaporated to give LA-4-e (11 mg, crude) as a yellow oil.

[0718] LCMS (ESI): m / z, 1537.1 [M+H] + Rt = 2.44 min.

[0719] Step 5: Preparation of compound LA-4

[0720] To a solution of compound LA-4-e (10 mg, 6.14 μmol, 1 eq), LY-82 (13.04 mg, 18.43 μmol, 3 eq) in DMF (3 mL) was added DIPEA (5.56 mg, 43.00 μmol, 7 eq) and HATU (7.01 mg, 18.43 μmol, 3 eq) at 0 °C, after addition, the reaction was stirred at 25 °C for 2 h, LCMS showed the reaction was completed, the reaction was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150 x 21.2 mm x 5 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 21% to 51% elution), and then lyophilized to give compound LA-4 (4.80 mg, yield 37.40%) as a white solid.

[0721] LCMS (ESI): m / z, 1045.8 [1 / 2 M+Na] + .

[0722] 1H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 8.17 (d, J = 8.0 Hz, 2H), 7.94 (d, J = 7.5 Hz, 1H), 7.80 (s, 1H), 7.72 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 8.3 Hz, 2H), 7.27 (d, J = 8.3 Hz, 2H), 7.09 (s, 1H), 7.00 (s, 2H), 6.20 (d, J = 3.0 Hz, 1H), 6.01 - 5.92 (m, 2H), 5.42 (s, 2H), 4.94 (s, 3H), 4.87 (d, J = 3.3 Hz, 2H), 4.81 (s, 1H), 4.75 (s, 1H), 4.61 (t, J = 6.9 Hz, 2H), 4.48 (d, J = 5.5 Hz, 2H), 4.32 (dd, J = 11.8, 6.1 Hz, 6H), 4.28 - 4.14 (m, 4H), 4.00 (s, 2H), 3.88 - 3.65 (m, 8H), 3.58 (q, J = 5.8, 4.7 Hz, 7H), 3.48 (d, J = 8.1 Hz, 14H), 3.41 - 3.35 (m, 15H), 3.19 (s, 5H), 2.97 (dd, J = 12.6, 6.7 Hz, 8H), 2.81 - 2.72 (m, 3H), 2.68 - 2.57 (m, 5H), 2.29 (t, J = 6.6 Hz, 3H), 2.16 - 2.04 (m, 4H), 2.01 - 1.88 (m, 4H), 1.76 - 1.56 (m, 10H), 1.47 (h, J = 7.3 Hz, 9H), 1.33 (dt, J = 16.7, 8.3 Hz, 7H), 1.22 - 1.04 (m, 5H), 1.03 (s, 3H), 0.83 (dd, J = 18.6, 6.7 Hz, 6H).

[0723] Example 11: Preparation of compound LA-6

[0724] Step 1: Preparation of compound LA-6-a

[0725] To a solution of compound LY-10a (purchased from Bide, 300 mg, 679.47 pmol, 1 eq) in MeOH (10 mL) was added LY-35i (1.94 g, 16 eq) and triethylamine (411 mg, 6 eq) at room temperature, then the reaction was heated to 70 °C for 21 h. LCMS showed the reaction was completed. The reaction solution was rotary evaporated to get the crude product. The crude product was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150 x 21.2 mm x 5 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 11% to 41% elution). After lyophilization, LA-6-a (354 mg, yield 84.08%) was obtained as a colorless oil.

[0726] LCMS (ESI): m / z, 620.4 [M+H] + Rt = 2.187 min.

[0727] Step 2: Preparation of compound LA-6

[0728] To a solution of compound LA-6-a (3 mg, 1.98 pmol, 1 eq), LA-4-e (13.04 mg, 18.43 pmol, 3 eq) (the synthesis of this compound please refer to the example of LA-4) in DMF (3 mL) was added DIPEA (7.38 mg, 6 eq) and HATU (6.77 mg, 9 eq) at 0 °C. After the addition was completed, the reaction was heated to 25 °C for 2 h. LCMS showed that the reaction was completed. The reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150 x 21.2 mm x 5 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 12% to 42% elution). After lyophilization, compound LA-6 (0.360 mg, yield: 8.59%) was obtained as a white solid.

[0729] LCMS (ESI): m / z, 1058.3 [1 / 2 M+Na] + .

[0730] 1H NMR (400 MHz, DMSO-d6) δ 9.95 (s, 1H), 8.45 (s, 1H), 8.18 (s, 2H), 7.94 (d, J = 7.5 Hz, 1H), 7.80 (s, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.58 (d, J = 8.2 Hz, 3H), 7.27 (d, J = 8.2 Hz, 2H), 7.09 (s, 1H), 7.00 (s, 2H), 6.19 (d, J = 2.9 Hz, 1H), 5.99 (s, 1H), 5.94 (d, J = 2.9 Hz, 1H), 5.43 (d, J = 4.4 Hz, 3H), 4.94 (s, 3H), 4.87 (d, J = 3.4 Hz, 2H), 4.78 (d, J = 25.9 Hz, 3H), 4.61 (t, J = 6.4 Hz, 2H), 4.55 (d, J = 3.9 Hz, 1H), 4.48 (s, 1H), 4.41 (d, J = 7.2 Hz, 1H), 4.33 (dd, J = 13.1, 7.6 Hz, 4H), 4.25 - 4.14 (m, 3H), 4.08 - 3.97 (m, 3H), 3.87 (d, J = 26.7 Hz, 4H), 3.73 (d, J = 5.5 Hz, 4H), 3.57 (t, J = 6.6 Hz, 5H), 3.53 - 3.45 (m, 28H), 3.19 (s, 5H), 3.06 - 2.89 (m, 10H), 2.82 - 2.71 (m, 4H), 2.28 (t, J = 6.6 Hz, 5H), 2.10 (dt, J = 12.3, 6.6 Hz, 4H), 1.97 (d, J = 15.5 Hz, 4H), 1.84 (d, J = 11.3 Hz, 2H), 1.70 (d, J = 13.3 Hz, 11H), 1.53 - 1.40 (m, 9H), 1.38 - 1.26 (m, 7H), 1.18 (q, J = 7.7 Hz, 3H), 1.08 - 0.99 (m, 4H), 0.83 (dd, J = 18.4, 6.7 Hz, 6H).

[0731] Example 12: Preparation of compound LA-9

[0732] To a solution of compound PE-1 (8 mg, 8.55 mmol, 1 eq), (6-(2,5-dioxo-2,5-dihydro-1H- pyrrol-1-yl)hexanoyl)glycyl-L-phenylalanine glycine (LA-9-b, 4.53 mg, 8.55 mmol, 1 eq, purchased from Haoyuan) in N,N-dimethylformamide (2 mL) was added N,N- diisopropylethylamine (2.83 μL, 17.1 μmol, 2 eq) and N,N,N',N'-tetramethyl-O-(7- azabenzotriazol-1-yl)uronium hexafluorophosphate (3.9 mg, 10.26 μmol, 1.2 eq) under ice bath, stirred for 5 min, then stirred at room temperature for 2 h. LCMS showed the starting material was consumed completely. The reaction solution was prepared by high performance liquid chromatography (preparative chromatography manufacturer: Orange SHIMADZU, model LC-20AP. The chromatographic column was GS-120-10-C18AP. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to obtain the product LA-9 (0.89 mg, yield 7.43%). LCMS (ESI): m / z, 1400 [M+H] + .

[0733] Example 13: Preparation of compound LA-10

[0734] Step 1: Preparation of compound LA-10a

[0735] Compound eribulin mesylate (40 mg, 48.43 μmol, 1 eq), (((9H-fluoren-9-yl)methoxy) carbonyl)-L-serine (19.02 mg, 58.11 μmol, 1.2 eq) were dissolved in DMF (3 mL), N,N- diisopropylethylamine (12.52 mg, 16.01 μL, 96.85 μmol, 2 eq), N,N,N',N'-tetramethyl-O-(7- azabenzotriazol-1-yl)uronium hexafluorophosphate (22.10 mg, 58.11 μmol, 1.2 eq) were added sequentially at 0 °C. The solution was stirred at 10 °C for 2 h. LCMS showed the starting material was consumed completely, and the compound LA-10a (50 mg, crude without yield) was obtained by oil pump concentration.

[0736] LCMS (ESI): m / z, 1039.4 [M+H] + .

[0737] Step 2: Preparation of compound LA-10b

[0738] Compound LA-10a (crude, 50 mg, 48.11 μmol, 1 eq) was dissolved in N,N- dimethylformamide (2 mL), 1,8-diazobicyclo[5.4.0]undec-7-ene (21.97 mg, 21.56 μL, 144.34 μmol, 3 eq) was added, and the reaction was stirred at 10 °C for 2 hours. LCMS showed that the starting material was completely consumed. The reaction solution was used directly in the next step without further treatment.

[0739] LCMS (ESI): m / z, 817.4 [M+H] + .

[0740] Step 3: Preparation of compound LA-10

[0741] Compound LA-10b was dissolved in N,N-dimethylformamide (2 mL), (6-(2,5-dioxo- 2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylglycyl-L-phenylalanylglycine (27.22 mg, 51.41 μmol, 1.2 eq) and 4-hydroxybenzotriazole (8.68 mg, 5.79 μL, 64.26 μmol, 1.5 eq) were added, followed by the addition of 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (12.32 mg, 14.05 μL, 64.26 μmol, 1.5 eq), and the reaction was stirred at 10 °C for 1 hour. LCMS showed that the reaction was complete. The reaction solution was prepared into compound LA-10 (40.14 mg, yield 69.52%, purity 98.57%) by high performance liquid chromatography.

[0742] The high performance liquid chromatography preparation conditions were as follows:

[0743] The preparation chromatography manufacturer was Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 50 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%.

[0744] LCMS (ESI): m / z, 1328.5 [M+H] + .

[0745] 1H NMR (400 MHz, DMSO-d6) δ 8.28 (t, J = 5.7 Hz, 1H), 8.15 (d, J = 7.9 Hz, 1H), 8.06 (t, J = 5.7 Hz, 1H), 7.99 (t, J = 5.7 Hz, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.79 (t, J = 5.7 Hz, 1H), 7.25 (d, J = 5.3 Hz, 4H), 7.21 - 7.14 (m, 1H), 7.00 (s, 2H), 5.05 (s, 1H), 5.00 (s, 1H), 4.89 (t, J = 5.6 Hz, 1H), 4.83 (s, 1H), 4.75 (s, 1H), 4.66 - 4.59 (m, 2H), 4.55 (t, J = 4.2 Hz, 1H), 4.48 (td, J = 9.4, 8.9, 4.3 Hz, 1H), 4.27 (t, J = 10.7 Hz, 2H), 4.17 (td, J = 10.3, 4.4 Hz, 1H), 4.10 (s, 3H), 4.02 (s, 1H), 3.84 - 3.64 (m, 8H), 3.62 - 3.46 (m, 6H), 3.26 (s, 4H), 3.11 - 3.00 (m, 3H), 2.87 - 2.63 (m, 5H), 2.60 - 2.54 (m, 1H), 2.36 - 2.17 (m, 6H), 2.16 - 2.07 (m, 3H), 1.97 (d, J = 33.5 Hz, 6H), 1.76 - 1.53 (m, 7H), 1.47 (h, J = 6.8 Hz, 6H), 1.29 (d, J = 13.1 Hz, 3H), 1.20 (d, J = 7.4 Hz, 3H), 1.00 (dd, J = 22.7, 9.1 Hz, 4H).

[0746] Example 14: Preparation of compound LA-11

[0747] Compound PE-10 (4 mg, 4.29 pmol, 1 eq) was dissolved in DMF (1 mL), (6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanoyl)glycylglycyl-L-phenylalanine glycine (LY-33b, purchased from Haoyuan Pharm, 2.27 mg, 4.29 pmol, 1 eq) and HOBT (1-hydroxybenzotriazole, 868.80 pg, 5.79 e -1 mL, 6.43 pmol, 1.5 eq), then EDCI.HC1 (1.23 mg, 1.41 pL, 6.43 pmol, 1.5 eq) was added, stirred at 15 °C for 1 h. LCMS showed product was generated. The reaction solution was prepared by high performance liquid chromatography to obtain the product (8.20 mg, yield 70.91%) as a white solid.

[0748] The high performance liquid chromatography preparation conditions are as follows:

[0749] The preparation chromatograph is Oriendo of the manufacturer, model BRIX-2860. The chromatographic column is Phenomenex Luna C18 250x30mmx10um. The mobile phase is water (0.225% HCOOH)-acetonitrile, and the elution proportion of acetonitrile is from 30% to 60% elution.

[0750] LCMS (ESI): m / z, 1444.7 [M+H] + .

[0751] Example 15: Preparation of compound LA-14

[0752] Step 1: Preparation of compound LA-14-b

[0753] Compound eribulin mesylate (10 mg, 12.11 umol, 1 eq) was dissolved in DMF (2 mL), (1R, 4R)-4-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid (purchased from Haoyuan Pharmaceutical, 3.53 mg, 14.53 umol, 1.2 eq) and DIPEA (3.13 mg, 4 uL, 24.21 umol, 2 eq) were added, HATU (5.52 mg, 14.53 umol, 1.2 eq) was added at 0°C, and stirred at 25°C for 1 hour. LCMS showed that the product was generated. The reaction solution was prepared by high performance liquid chromatography to obtain white solid product (8.20 mg, yield 70.91%).

[0754] The high performance liquid chromatography preparation conditions are as follows:

[0755] The preparation chromatograph is Oriendo of the manufacturer, model BRIX-2860. The chromatographic column is Welch Xtimate C18 150x30mmx10um. The mobile phase is water (0.225% HCOOH)-acetonitrile, and the elution proportion of acetonitrile is from 30% to 60% elution.

[0756] LCMS (ESI): m / z, 977.6 [M+Na] + .

[0757] Step 2: Preparation of compound PE-17

[0758] Compound LA-14-b (8.20 mg, 8.58 pmol, 1 eq) was dissolved in DCM (2 mL), TFA (307 mg, 200 pL, 2.69 mmol, 313.6 eq) was added under ice-bath, and the mixture was stirred at 0-25 °C for 12 h. LCMS showed the reaction was complete. The reaction solution was directly rotary-evaporated to dryness to give the product (7 mg, yield 95.36%) as white oil, which was used directly in the next step.

[0759] LCMS (ESI): m / z, 855.5 [M+H] + .

[0760] Step 3: Preparation of compound LA-14

[0761] Compound PE-17 (5 mg, 5.85 pmol, 1 eq) was dissolved in DMF (1 mL), DIPEA (2.27 mg, 2.90 pL, 17.54 pmol, 3 eq) and compound BL-2-a (purchased from Haoyuan, 5.18 mg, 7.02 pmol, 1.2 eq) were added at 0 °C, and the mixture was stirred at 25 °C for 12 h. LCMS showed the starting material was consumed completely. The reaction solution was purified by high performance liquid chromatography to give the product LA-14 (8 mg, yield 86.81%) as white solid.

[0762] The high performance liquid chromatography preparation conditions were as follows:

[0763] The high performance liquid chromatography preparation conditions were as follows:

[0764] LCMS (ESI): m / z, 1476.8 [(M+H)] + .

[0765] Example 16: Preparation of compound LA-15

[0766] To a solution of PE-18 (2.05 mg, 100%, 2.36 pmol, 1 eq) and N, N- dimethylformamide (0.5 mL) at 0 °C, add BL-2-a (purchased from Haoyuan Pharmatech, 2.09 mg, 2.84 pmol, 1.2 eq) and N, N- diisopropylethylamine (916.66 pg, 1.17 pL, 7.09 pmol, 3 eq), stir at 20 °C for 1 h, LCMS shows the starting material is consumed completely. The reaction solution is prepared by high performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. Column: Welch Xtimate C18 250*30mm*10pm. Mobile phase: water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 50% to 80% elution), and white solid compound LA-15 (700 pg, 0.46 pmol, yield 19.42%, purity 96.15%) is obtained after lyophilization.

[0767] LCMS (ESI): m / z, 733.6 [1 / 2 M+H] + .

[0768] Example 17: Preparation of compound LA-21

[0769] Step 1: Preparation of compound LA-21-a

[0770] Compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq) is dissolved in DMF (1 mL), add 2-(tert-butoxycarbonyl)-2-azaspiro[3.5]nonane-7-carboxylic acid (3.69 mg, 13.70 pmol, 1 eq) and DIPEA (3.54 mg, 4.53 pL, 27.40 pmol, 2 eq), add HATU (6.25 mg, 16.44 pmol, 1.2 eq) at 0 °C, stir at 0-25 °C for 2 h. LCMS shows that the product is generated. The reaction solution is prepared by high performance liquid chromatography to obtain white solid product LA-21-a (7 mg, yield 52.07%).

[0771] High performance liquid chromatography preparation conditions are as follows:

[0772] Preparative chromatography manufacturer Shimadzu, model LC-20AP. Column: Welch Xtimate C18 250*30mm*10pm. Mobile phase: water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 50% to 80% elution.

[0773] LCMS (ESI): m / z, 981.6 [M+H] + .

[0774] Step 2: Preparation of compound LA-21-b

[0775] LA-21-a (7 mg, 7.13 μmol, 1 eq) was dissolved in DCM (2 mL), TFA (307 mg, 200 μL, 2.69 mmol, 377.4261 eq) was added at 0 °C, and the mixture was stirred at 25 °C for 0.5 h. LCMS showed that the reaction was complete. The reaction solution was directly rotary evaporated to give the product LA-21-b (5 mg, yield 79.54%) as yellow oil.

[0776] LCMS (ESI): m / z, 881.6 [M+H] + .

[0777] Step 3: Preparation of compound LA-21

[0778] LA-21-b (6 mg, 1 eq) was dissolved in DMF (2 mL) at room temperature, DIPEA (1 mg, 1.28 μL, 7.74 μmol, 3 eq) was added, and BL-2-a (8 mg, 10.84 μmol, 1.2 eq, purchased from Haoyuan) was added at 0 °C. The reaction solution was stirred at 25 °C for 1 h. LCMS showed that the starting material was completely reacted. The reaction solution was purified by high performance liquid chromatography to give the product LA-21 (880 μg, purity 80%) as a white solid.

[0779] The high performance liquid chromatography preparation conditions were as follows:

[0780] The high performance liquid chromatography preparation conditions were as follows:

[0781] LCMS (ESI): m / z, 1502.9 [(M+H)] + .

[0782] Example 18: Preparation of compound LA-22

[0783] Step 1: Preparation of tert-butyl 9-(((4-nitrophenoxy)carbonyl)amino)-3- azaspiro[5.5]undecane-3-carboxylate

[0784] To a solution of compound 9-amino-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (purchased from Dookyou, 30 mg, 111.78 pmol, 1 eq) in DMF (1 mL) was added bis(4-nitrophenyl) carbonate (102.01 mg, 68.01 pL, 335.33 pmol, 3 eq) and DIPEA (28.89 mg, 36.95 pL, 223.56 pmol, 2 eq) at 0 °C, and the mixture was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction solution was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Phenomenex Luna C18 150 x 25 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution), and then lyophilized to give 9-(((4-nitrophenoxy)carbonyl)amino)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (19.50 mg, 40.24% yield).

[0785] LCMS (ESI): m / z, 334 [M-100+H] + Rt = 1.082 min.

[0786] Step 2: Preparation of compound LA-22-a

[0787] To a solution of compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq), 9-(((4-nitrophenoxy)carbonyl)amino)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (7.35 mg, 16.95 pmol, 1.4 eq) in DMF (1 mL) was added HOBt (817.91 pg, 5.45 e -1 Rt = 3.55 min.

[0788] LCMS (ESI): m / z, 1024.7 [M+H] + Rt = 3.55 min.

[0789] Step 3: Preparation of compound LA-22-b

[0790] To a solution of compound LA-22-a (14 mg, 13.67 pmol, 1 eq) in DCM (2 mL) was added TFA (0.2 mL) at 0 °C, after addition, the reaction solution was reacted at 0-10 °C for 1 h, LCMS showed that the reaction was completed, the reaction solution was directly rotary dried to obtain yellow solid compound LA-22-b (15 mg, crude).

[0791] LCMS (ESI): m / z, 946.7 [M+Na] + Rt = 2.49 min.

[0792] Step 4: Preparation of compound LA-22

[0793] To a solution of compound LA-22-b (15 mg, 16.23 pmol, 1 eq) and BL-2-a (11.97 mg, 16.23 pmol, 1 eq) in DMF (2 mL) was added DIPEA (6.29 mg, 8.05 pL, 48.69 pmol, 3 eq) at 0 °C, after addition, the reaction was reacted at 25 °C for 1 h, LCMS showed that the reaction was completed, the reaction solution was freeze-dried by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150*30 mm*10 pm. The mobile phase was water (10 mM NH4HCO3)-acetonitrile, and the elution ratio of acetonitrile was from 50% to 80% elution) to obtain white solid compound LA-22 (2.96 mg, yield 11.98%, purity 84%).

[0794] LCMS (ESI): m / z, 1544.9 [M+Na] + , 761.9 [1 / 2M+1] + Rt = 2.44 min.

[0795] Example 19: Preparation of compound LA-23

[0796] Step 1: Preparation of tert-butyl 9-(((4-nitrophenoxy)carbonyl)oxy)-3- azaspiro[5.5]undecane-3-carboxylate

[0797] To a solution of compound 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (purchased from Dookere, 50 mg, 185.61 pmol, 1 eq) in DMF (3 mL) was added bis(4-nitrophenyl) carbonate (67.76 mg, 45.17 pL, 222.73 pmol, 1.2 eq) and DIPEA (47.98 mg, 61.35 pL, 371.22 pmol, 2 eq) at 0 °C. After addition, the mixture was stirred at 30 °C for 1 h. LCMS showed the reaction was completed. The reaction solution was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 60% to 90% elution), and then lyophilized to give 9-(((4-nitrophenoxy)carbonyl)oxy)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (26.90 mg, yield 33.36%) as a light yellow solid.

[0798] LCMS (ESI): m / z, 335 [M-100+H] + Rt = 3.459 min.

[0799] Step 2: Preparation of compound LA-23-a

[0800] To a solution of compound eribulin mesylate (10 mg, 13.70 pmol, 1 eq), 9-(((4-nitrophenoxy)carbonyl)oxy)-3-azaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (8.33 mg, 19.18 pmol, 1.4 eq) in DMF (3 mL) was added HOBt (925.61 pg, 6.17 e -1 After addition, the mixture was stirred at 25 °C for 3 h. LCMS showed the reaction was completed. The reaction solution was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 60% to 90% elution), and then lyophilized to give compound LA-23-a (12 mg, yield 85.43%) as a white solid.

[0801] LCMS (ESI): m / z, 1047.6 [M+Na] + Rt = 3.206 min.

[0802] Step 3: Preparation of compound LA-23-b

[0803] To a solution of compound LA-23-a (12 mg, 11.70 pmol, 1 eq) in DCM (6 mL) was added TFA (0.6 mL) at 0 °C. The reaction was stirred at 0-10 °C for 30 min. LCMS showed the reaction was completed. The reaction was directly rotary-evaporated to dryness to give compound LA-23-b (11 mg, crude) as a yellow solid.

[0804] LCMS (ESI): m / z, 947.6 [M+Na] + Rt = 1.947 min.

[0805] Step 4: Preparation of compound LA-23

[0806] To a solution of compound LA-23-b (11 mg, 11.89 pmol, 1 eq) and BL-2-a (11.97 mg, 16.23 pmol, 1 eq) in DMF (2 mL) was added DIPEA (4.61 mg, 5.89 pL, 35.67 pmol, 3 eq) at 0 °C. The reaction was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction was directly freeze-dried to give compound LA-23 (8.84 mg, yield 48.79%, purity 84%) as a white solid by high performance liquid chromatography preparation (Prep-chromatography factory Oriendo, model BRIX-2860. The column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% FA) - acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution).

[0807] LCMS (ESI): m / z, 1546.8 [M+Na] + , 785.2 [1 / 2 M+Na] + Rt = 2.564 min.

[0808] Example 20: Preparation of compound LA-24

[0809] Step 1: Preparation of (9H-fluoren-9-yl)methyl ((S)-1-(((S)-1-(2- hydroxyethoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1- oxobutan-2-yl)carbamate

[0810] To a solution of compound LY-27a (250 mg, 519.16 pmol, 1 eq) in THF (10 mL) was added 2-((tert-butyldimethylsilyl)oxy)ethan-1-ol (183.09 mg, 1.04 mmol, 2 eq) and TsOH (26.82 mg, 25.07 pL, 155.75 pmol, 0.3 eq) at room temperature. After addition, the reaction was stirred at 33 °C for 3 h. LCMS showed the reaction was completed. The reaction was concentrated to get the crude product. The crude product was purified by normal reverse silica gel column (ISCO, 220 g SepaFlash Silica Flash Column, Eluent of 0-47% water / CH3CN, 100 mL / min). The yellow solid compound LY-22AD-a (98 mg, yield 39.04%) was obtained by lyophilization.

[0811] LCMS (ESI): m / z, 506.3 [M+H] + Rt = 1.813 min.

[0812] Step 2: Preparation of compound LY-22AD-b

[0813] To a solution of compound LY-22AD-a (98 mg, 202.66 pmol, 1 eq) in DMF (3 mL) was added di(p-nitrophenyl) carbonate (123.30 mg, 82.20 pL, 405.32 pmol, 2 eq) and DIPEA (78.58 mg, 100.48 pL, 607.98 pmol, 3 eq) at 0 °C. After addition, the reaction was stirred at 33 °C for 2 h. LCMS showed the reaction was completed. The reaction was concentrated to get the crude product. The crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane 1 :20 to 1 :10) to get the yellow solid compound LY-22AD-b (111 mg, crude, yield 75.99%, purity 90%).

[0814] Step 3: Preparation of compound LA-24-a

[0815] To a solution of compound Nintedanib mesylate (10 mg, 12.11 pmol, 1 eq) in DMAC (3 mL) was added DIPEA (1.56 mg, 2 pL, 12.11 pmol, 1 eq) at 0 °C, then stirred at 25 °C for 10 min, then added LY-22AD-b (8.73 mg, 90%, 12.11 pmol, 1 eq), HOBt (817.91 pg, 6.05 pmol, 0.5 eq) and pyridine (4.79 mg, 4.90 pL, 60.53 pmol, 5 eq) to the reaction solution. After addition, the reaction was carried out at 25 °C for 2 h, LCMS showed that the reaction was completed, the reaction solution was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 50% to 80% elution), and the yellow solid compound LA-24-a (9.10 mg, yield 60.64%) was obtained by lyophilization.

[0816] LCMS (ESI): m / z, 1261.7 [M+Na] + Rt = 2.909 min.

[0817] Step 4: Preparation of compound LA-24-b

[0818] To a solution of compound LA-24-a (9.10 mg, 7.34 pmol, 1 eq) in DMF (2 mL) was added DBU (3.35 mg, 3.29 pL, 22.03 pmol, 3 eq) at 0 °C, after addition, the reaction was carried out at 25 °C for half an hour, LCMS showed that the reaction was completed, TsOH (3.79 mg, 22.03 pmol, 3 eq) was added to the reaction solution, then the reaction solution was directly used for the next step.

[0819] LCMS (ESI): m / z, 1017.7 [M+H] + Rt = 1.629 min.

[0820] Step 5: Preparation of compound LA-24

[0821] To a solution of compound LA-24-b (7.50 mg, 7.37 pmol, 1 eq) and LY-22CD-3-d (5.88 mg, 7.37 pmol, 1 eq) in DMF (2 mL) was added DIPEA (3.81 mg, 4.87 pL, 29.49 pmol, 4 eq) and HATU (3.36 mg, 8.85 pmol, 1.2 eq) at 0 °C. After 25 °C for half an hour, LCMS showed the reaction was completed. The reaction mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 30% to 60%), and white solid compound LA-24 (3.55 mg, yield 26.81%, purity 100%) was obtained by lyophilization.

[0822] LCMS (ESI): m / z, 1819.0 [M+Na] + , 899.4 [1 / 2M+1] + Rt = 2.069 min.

[0823] Example 21: Preparation of compound LA-25A

[0824] Step 1: Preparation of compound LA-25-a

[0825] To a solution of compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq) and DIPEA (2 pL, 12 pmol, 1 eq) in DMAC (2 mL) was added compound LY-22CC-c (12 mg, 18 pmol, 1.5 eq), HOBT (0.8 mg, 6 pmol, 0.5 eq), pyridine (4.90 pL, 60 pmol, 5 eq) at room temperature. The reaction mixture was stirred at 25 °C for 12 hours. LCMS showed the reaction was completed. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860-R1. The chromatographic column was Phenomenex Luna C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was eluted from 50% to 80%), and white solid LA-25-a (15 mg, yield 97.75%) was obtained.

[0826] Step 2: Preparation of compound LA-25-b

[0827] Compound LA-25-a (2.40 g, 4.65 mmol, 1 eq) was dissolved in DMF (2 mL), then DBU (15 mg, 12 pmol, 1 eq) was added, the reaction was stirred at 25 °C for half an hour. LCMS showed that the reaction was complete. To the reaction solution, 15 mg of TsOH was added to quench the reaction, then the reaction solution was directly used for the next step (12.37 mg, crude in 2 mL DMF).

[0828] Step 3: Preparation of compound LA-25A

[0829] To the solution of compound LA-25-b (12.37 mg) in DMF (2 mL) was added LY-22CD-3-d (9.43 mg, 11.83 pmol, 1 eq), N,N-diisopropylethylamine (7.82 pL, 47 pmol, 4 eq) and HATU (5.40 mg, 14.20 pmol, 1.2 eq) at 0 °C, the mixture was stirred at room temperature for half an hour, LCMS showed that the starting material was completely reacted. The mixture was prepared by high performance liquid chromatography (Oriendo, model BRIX-2860-R1. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was from 30% to 60% elution), to obtain white solid LA-25A (2.50 mg, yield 11.58%).

[0830] LCMS (ESI): m / z, 865.4 [M+H] + .

[0831] Example 22: Preparation of compound LA-25AA

[0832] Step 1: Preparation of compound LA-25AA-a

[0833] To a solution of compound LA-27AA-a (50 mg, 63.79 pmol, 1 eq, the synthesis of this compound please refer to the preparation of BA-1-c in example 1) and 5-amino-1-pentanol (5.69 mg, 63.79 pmol, 1 eq) in DMF (2 mL) was added DIPEA (16.49 mg, 127.59 pmol, 2 eq) at 0 °C. The reaction was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860-R1. The chromatographic column was Welch Xtimate C18 250*30 mm*10 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 10% to 40% elution), to get white solid LA-25AA-a (33 mg, yield 70.49%).

[0834] Step 2: Preparation of compound LA-25AA-b

[0835] To a solution of compound LA-25AA-a (33 mg, 44.97 pmol, 1 eq) in DMF (2 mL) was added NPC (82.08 mg, 269.82 pmol, 6 eq) and DIPEA (34.87 mg, 269.82 pmol, 6 eq), the reaction was stirred at 25 °C for 1 hour. LCMS showed that the reaction was completed. The reaction was dried under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane 0:1 to 1:10) to obtain white solid product LA-25AA-b (31 mg, yield 76.69%).

[0836] Step 3: Preparation of compound LA-25AA

[0837] To a solution of compound LA-25AA-b (7.18 mg, 7.99 pmol, 1.1 eq) and eribulin mesylate (6 mg, 7.26 pmol, 1 eq) in DMF (2 mL) was added N,N-diisopropylethylamine (2.82 mg, 21.79 pmol, 3 eq) at 0 °C, the mixture was stirred at room temperature for 2 hours, LCMS showed that the raw material was completely reacted. The mixture was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150*30 mm*10 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, acetonitrile elution ratio from 30% to 60% elution), to get white solid LA-25AA (5.80 mg, yield 53.60%).

[0838] LCMS (ESI): m / z, 1512.8 [M+Na] + .

[0839] Example 23: Preparation of compound LA-25B

[0840] Compound LY-29C-c (4.73 mg, 7.65 μmol, 1 eq) was dissolved in DMF (500 μL) and cooled to 0 °C, then added compound LA-25B-b (8 mg, 7.65 μmol, 1 eq), HATU (2.91 mg, 7.65 μmol, 1 eq) and DIPEA (2.97 mg, 3.79 μL, 22.96 μmol, 3 eq). The reaction was reacted at 25 °C for 1 hour. LCMS showed the reaction was complete. The reaction was prepared by high performance liquid chromatography (Prep-Chromatography Factory Oriendo, Model BRIX-2860. The column was Phenomenex Luna C18 250 x 50 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution), to get white solid LA-25B (1.98 mg, purity 100%, yield 15.73%).

[0841] LCMS (ESI): m / z, 1667.2 [M+Na] + , Rt = 2.576 min.

[0842] Example 24: Preparation of compound LA-26

[0843] Step 1: Preparation of compound LA-26-b

[0844] Compound LY-27a (250 mg, 519 μmol, 1 eq) was dissolved in THF (3 mL) and cooled to 0 °C, then added S-1,2-propanediol (118.5 mg, 1.56 mmol, 3 eq) and TsOH (89.4 mg, 519 μmol, 1 eq). The reaction was reacted at 35 °C for 3 hours. LCMS showed the reaction was complete. The reaction was concentrated to get white solid 500 mg, then prepared by high performance liquid chromatography (Prep-Chromatography Factory Oriendo, Model BRIX-2860. The column was Phenomenex Luna C18 250 x 50 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution), to get white solid LA-26-b (120 mg, yield 46.45%).

[0845] LCMS (ESI): m / z, 520.4 [M+Na] + Rt = 1.914 min.

[0846] Step 2: Preparation of compound LA-26-c

[0847] Compound LA-26-b (99.5 mg, 200 pmol, 1 eq) was dissolved in DMF (2 mL), cooled to 0 °C, then added NPC (121.6 mg, 400 pmol, 2 eq) and DIPEA (77.5 mg, 99.1 pL, 600 pmol, 3 eq). The reaction was reacted at 25 °C for 2 hours. LCMS showed the reaction was complete. The reaction was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 50 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution), to get white solid LA-26-c (16 mg, yield 12.1%).

[0848] LCMS (ESI): m / z, 685.3 [M+Na] + Rt = 3.889 min.

[0849] Step 3: Preparation of compound LA-26-d

[0850] Compound LA-26-c (3.98 mg, 6 pmol, 1 eq) was dissolved in DMAC (500 pL), cooled to 0 °C, then added DIPEA (2.33 mg, 2.97 pL, 18 pmol, 3 eq) and eribulin mesylate (4.96 mg, 6 pmol, 1 eq). The reaction was reacted at 25 °C for 3 hours. LCMS showed the reaction was complete. The reaction was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Phenomenex Luna C18 250 x 50 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution), to get white solid LA-26-d (7 mg, yield 93.1%).

[0851] LCMS (ESI): m / z, 276.7 [M+Na] + Rt = 3.023 min.

[0852] Step 4: Preparation of compound LA-26-e

[0853] Compound LA-26-d (7 mg, 5.58 pmol, 1 eq) was dissolved in DMF (500 pL), cooled to 0 °C, then added DBU (2.55 mg, 2.50 pL, 16.75 pmol, 3 eq). The reaction was reacted at 25 °C for 1 h. LCMS showed the reaction was complete. TsOH (2.88 mg, 16.75 pmol, 3 eq) was added to the reaction, and reacted for 2 min. The reaction was used directly for the next step without further workup.

[0854] LCMS (ESI): m / z, 1031.7 [M+H] + , Rt = 1.705 min.

[0855] Step 5: Preparation of compound LA-26

[0856] Compound LA-26-e (5 mg, 4.85 pmol, 1 eq) was dissolved in DMF (500 pL), cooled to 0 °C, then added LY-22CD-3-d (3.86 mg, 4.85 pmol, 1 eq), DIPEA (1.88 mg, 2.40 pL, 14.55 pmol, 3 eq) and HATU (1.84 mg, 4.85 pmol, 1 eq). The reaction was reacted at 25 °C for 1 h. LCMS showed the reaction was complete. The reaction was prepared by high performance liquid chromatography (Prep-Chrom Oriendo, Model BRIX-2860. The column was Phenomenex Luna C18 250 x 50 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution), to obtain LA-26 (5.37 mg, yield 61.2%, purity 98.57%) as a white solid.

[0857] LCMS (ESI): m / z, 1832.1 [M+Na] + , Rt = 2.142 min.

[0858] Example 25: Preparation of compound LA-26B

[0859] Step 1: Preparation of compound LA-26B-b

[0860] To a solution of compound S-1, 2-propanediol (50 mg, 63.79 μmol, 1 eq) in THF (66 mL) was added 1H-imidazole (1.07 g, 976.07 μL, 15.77 mmol, 1.2 eq) and TBDPS-Cl (3.79 g, 3.58 mL, 13.80 mmol, 1.05 eq), and the reaction was stirred at room temperature for 20 hours. LCMS showed that the reaction was completed. The reaction solution was extracted with EA (30 mL x 3), and the combined organic phase was washed with H2O (20 mL x 2), dried over anhydrous sodium sulfate, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1:1) to give compound (2S)-1-((tert-butyldiphenylsilyl)oxy)propan-2-ol (3.70 g, yield 89.52%) as colorless oil.

[0861] 1 H NMR (400 MHz, Chloroform-d) δ 7.69 (dt, J = 8.0, 2.1 Hz, 4H), 7.49-7.35 (m, 6H), 3.96-3.93 (m, 1H), 3.64 (dd, J = 10.1, 3.5 Hz, 1H), 3.47 (dd, J = 10.1, 7.8 Hz, 1H), 1.12 (d, J = 6.3 Hz, 3H), 1.09 (s, 9H).

[0862] Step 2: Preparation of compound LA-26B-c

[0863] To a solution of compound (2S)-1-[(tert-butyldiphenylsilyl)oxy]propan-2-ol (2.61 g, 8.31 mmol, 2 eq) and methyl [(2S)-2-[(2S)-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-3-methylbutanamido]propanamido]acetate (2 g, 4.15 mmol, 1 eq) in THF (30 mL) was added TsOH (715.19 mg, 4.15 mmol, 1 eq), and the reaction was stirred at 30 °C for 2 hours. LCMS showed that the reaction was completed. The reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer ISCO, model LC-20AP. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of acetonitrile was from 65% to 95% elution) to give compound LA-26B-c (233 mg, yield 7.62%) as white solid. LCMS (ESI): m / z, 758 [M+Na] + .

[0864] Step 3: Preparation of compound LA-26B-d

[0865] To a solution of compound LA-26B-c (277.20 mg, 376.63 pmol, 1 eq) in THF (20 mL) was added Et3N.3HF (2.43 g, 2.46 mL, 15.07 mmol, 40 eq), 25 °C-70 °C for 2 hours, LCMS showed the reaction was completed, the residue was purified by silica gel column chromatography (C18, ISCO, R-120 g SepaFlash Silica Flash Col urn, Eluent of 10-40% water / CH3CN, 100 mL / min) to give compound LA-26B-d (130 mg, yield 69.37%, purity 100%) as a white solid. LCMS (ESI): m / z, 520 [M+Na] + .

[0866] Step 4: Preparation of compound LA-26B-e

[0867] To a solution of compound LA-26B-d (133.40 mg, 100%, 268.09 pmol, 1 eq) in DMF (2 mL) was added bis(4-nitrophenyl) carbonate (326.23 mg, 217.48 pL, 1.07 mmol, 4 eq) and DIPEA (207.89 mg, 265.84 pL, 1.61 mmol, 6 eq), 33 °C for 1 hour, LCMS showed the reaction was completed, the residue was purified by silica gel column chromatography (PE / EA from 10 / 1 to 4 / 1, DCM / MeOH = 10:1) to give compound LA-26B-e (197 mg, crude, purity 82%) as a white solid. LCMS (ESI): m / z, 685 [M+Na] + .

[0868] Step 5: Preparation of compound LA-26B-f

[0869] To a solution of LA-26B-e (12.18 mg, 82%, 15.07 pmol, 1.1 eq) and eribulin mesylate (10 mg, 13.70 pmol, 1 eq) in DMF (1 mL) was added DIPEA (5.31 mg, 6.79 pL, 41.10 pmol, 3 eq) and HOBT (1.85 mg, 13.70 pmol, 1 eq), after reaction at room temperature for 2 hours, LCMS showed the starting material disappeared, purified by high performance liquid chromatography (preparative chromatography manufacturer ISCO, model Lab311-DJ. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution) to give compound LA-26B-f (14 mg, yield 81.52%) as a white solid, LCMS (ESI): m / z, 1276 [M+Na] + .

[0870] Step 6: Preparation of compound LA-26B-g

[0871] To a solution of compound LA-26B-f (14 mg, 11.17 pmol, 1 eq) in DMF (1 mL) was added DBU (5.10 mg, 5.01 pL, 33.51 pmol, 3 eq), after reaction at room temperature for 40 minutes, LCMS showed the starting material disappeared, TsOH (5.77 mg, 33.51 pmol, 3 eq) was added, the reaction solution was used directly in the next step (theoretical yield 11.52 mg, crude product was not calculated yield), LCMS (ESI): m / z, 1053 [M+Na] + .

[0872] Step 7: Preparation of compound LA-26B

[0873] To a solution of compound LA-26B-g (11.52 mg, 11.17 μmol, 1 eq) and compound LY-22CD-3-d (8.90 mg, 11.17 μmol, 1 eq) in DMF (1 mL) was added DIPEA (2.89 mg, 3.69 μL, 22.34 μmol, 2 eq) and HATU (5.10 mg, 13.41 μmol, 1.2 eq) at 0 °C. After 1 h of reaction at room temperature, LCMS showed the starting material was consumed. Purification by high performance liquid chromatography preparation (preparative chromatography manufacturer ISCO, model BRIX-2860. The column was Synergi Max-RP 200 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) and lyophilization gave the product LA-26B (4 mg, yield 18.79%, purity 95%) as a white solid. LCMS (ESI): m / z, 1832 [M+Na] + .

[0874] Example 26: Preparation of compound LA-27A

[0875] Step 1: Preparation of compound LA-27-a

[0876] To a solution of compound LY-27a (200 mg, 415.33 μmol, 1 eq) in THF (8 mL) was added 5-((tert-butyldimethylsilyl)oxy)pentan-1-ol (181.42 mg, 830.65 μmol, 2 eq) and TsOH (21.46 mg, 20.05 μL, 124.60 μmol, 0.3 eq) at room temperature. After addition, the mixture was stirred at 33 °C for 3 h. LCMS showed the reaction was completed. The reaction was concentrated to give the crude product. The crude product was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer SHIMADZU, model LC-20AP. The column was Phenomenex Luna C18 150 x 25 mm x 10 μm. The mobile phase was water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution). Lyophilization gave compound LA-27-a (102 mg, yield 46.72%) as a yellow solid.

[0877] LCMS (ESI): m / z, 548.4 [M+Na] + . Rt = 2.009 min.

[0878] Step 2: Preparation of compound LA-27-b

[0879] To a solution of compound LA-27-a (102 mg, 194.05 pmol, 1 eq) in DMF (5 mL) was added di-(p-nitrophenyl) carbonate (118.06 mg, 388.09 pmol, 2 eq) and DIPEA (75.24 mg, 96.21 pL, 582.14 pmol, 3 eq) at 0 °C. After addition, the reaction was stirred at 33-35 °C for 3 h. LCMS showed the reaction was completed. The reaction was poured into 50 mL of ice water, extracted with ethyl acetate (40 mL x 3), the organic phase was combined and washed with water (60 mL x 2), brine (50 mL) successively, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by silica gel column chromatography (eluent: methanol / dichloromethane 1:20 to 1:10) to give compound LA-27-b (95 mg, crude yield 56.70%, purity 80%) as a yellow solid.

[0880] LCMS (ESI): m / z, 713.4 [M+Na] + Rt = 2.783 min.

[0881] Step 3: Preparation of compound LA-27-c

[0882] To a solution of compound eribulin mesylate (10 mg, 12.11 pmol, 1 eq) in DMAC (2 mL) was added DIPEA (1.56 mg, 2 pL, 12.11 pmol, 1 eq) at 0 °C, then stirred at 25 °C for 10 min, then LA-27-b (10.45 mg, 80%, 12.11 pmol, 1 eq), HOBt (817.91 pg, 6.05 pmol, 0.5 eq) and pyridine (4.79 mg, 4.90 pL, 60.53 pmol, 5 eq) were added. After addition, the reaction was stirred at 25 °C for 16 h. LCMS showed the reaction was completed. The reaction was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 250 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 50% to 80% elution) and lyophilized to give compound LA-27-c (11 mg, yield 70.90%) as a yellow solid.

[0883] LCMS (ESI): m / z, 1303.8 [M+Na] + Rt = 3.103 min.

[0884] Step 4: Preparation of compound LA-27-d

[0885] To a solution of compound LA-27-c (11 mg, 8.58 pmol, 1 eq) in DMF (2 mL) was added DBU (3.92 mg, 3.85 pL, 25.75 pmol, 3 eq) at 0 °C, after addition, the reaction solution was reacted at 25 °C for half an hour, LCMS showed that the reaction was completed, TsOH (4.43 mg, 25.75 pmol, 3 eq) was added to the reaction solution, and then the reaction solution was directly used for the next step.

[0886] LCMS (ESI): m / z, 1059.8 [M+H] + Rt = 1.700 min.

[0887] Step 5: Preparation of compound LA-27A

[0888] To a solution of compound LA-27-d (9 mg, 8.50 pmol, 1 eq) and LY-22CD-3-d (8.12 mg, 10.20 pmol, 1.2 eq) in DMF (2 mL) was added DIPEA (4.39 mg, 5.62 pL, 33.98 pmol, 4 eq) and HATU (3.88 mg, 10.20 pmol, 1.2 eq) at 0 °C. After addition, it was reacted at 25 °C for 20 minutes, LCMS showed that the reaction was completed, the reaction solution was prepared by high performance liquid chromatography (Prep HPLC, Oriendo, BRIX-2860. The chromatographic column was Welch Xtimate C18 150*30mm*10pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of acetonitrile was from 30% to 60% elution), and white solid compound LA-27A (8.92 mg, yield 57.12%, purity 98.54%) was obtained by freeze-drying.

[0889] LCMS (ESI): m / z, 1861.1 [M+Na] + Rt = 2.237 min.

[0890] Example 27: Preparation of compound LA-27AA

[0891] Step 1: Preparation of compound LA-27AA-b

[0892] To a solution of compound LA-27AA-a (50 mg, 63.79 pmol, 1 eq, please refer to the preparation of BA-1-c in example 1) and 5-amino-1-pentanol (6.58 mg, 63.79 pmol, 1 eq) in DMF (2 mL) was added DIPEA (8.24 mg, 63.79 pmol, 1 eq) at 0 °C. The reaction was stirred at room temperature for 1 h. LCMS showed the reaction was completed. The mixture was purified by high performance liquid chromatography preparation (Prep HPLC manufacturer Oriendo, model BRIX-2860-R1. The column was Welch Xtimate C18 250 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution) to give LA-27AA-b (28 mg, yield 58.69%) as a white solid.

[0893] Step 2: Preparation of compound LA-27AA-c

[0894] To a solution of compound LA-27AA-b (28 mg, 37.44 pmol, 1 eq) in DMF (2 mL) was added NPC (68.34 mg, 224.64 pmol, 6 eq) and DIPEA (29.03 mg, 224.64 pmol, 6 eq) and the reaction was stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction was rotary evaporated under reduced pressure and the residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane 0:1 to 1:10) to give LA-27AA-c (21 mg, yield 61.44%) as a white solid.

[0895] Step 3: Preparation of compound LA-27AA

[0896] To a solution of compound LA-27AA-c (7.29 mg, 7.99 pmol, 1.1 eq) and eribulin mesylate (6 mg, 7.26 pmol, 1 eq) in DMF (2 mL) was added N,N-diisopropylethylamine (2.82 mg, 21.79 pmol, 3 eq) at 0 °C. The mixture was stirred at room temperature for 2 h. LCMS showed the starting material was consumed completely. The mixture was purified by high performance liquid chromatography preparation (Prep HPLC manufacturer Oriendo, model BRIX-2860. The column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give LA-27AA (3.20 mg, yield 28.37%) as a white solid.

[0897] LCMS (ESI): m / z, 1526.9 [M+Na]+ .

[0898] Example 28: Preparation of compound LA-27AA-2

[0899] Step 1: Preparation of compound LA-27AA-2-a

[0900] To a solution of compound 5-amino-1-pentanol (55.29 mg, 535.98 umol, 1.1 eq) and N-[fluorenylmethoxycarbonyl]-L-valyl-L-alanine (200 mg, 487.26 umol, 1 eq) in DMF (8 mL) was added DIPEA (157.43 mg, 1.22 mmol, 2.5 eq) and HATU (222.33 mg, 584.71 umol, 1.2 eq) and stirred at room temperature for 2 hours, LCMS showed the starting material was consumed completely. The residue was purified by silica gel column chromatography (C18, ISCO, R-330g SepaFlash Silica Flash Column, Eluent of 10-50% water / CH3CN, 100 mL / min) to give LA-27AA-2-a (227.4 mg, yield 94.16%) as a white solid.

[0901] LCMS (ESI): m / z, 496 [M+H] + , Rt = 2.049 min.

[0902] Step 2: Preparation of compound LA-27AA-2-b

[0903] To a solution of LA-27AA-2-a (100 mg, 201.77 umol, 1 eq) in DMF (5 mL) was added DBU (92.15 mg, 605.30 umol, 3 eq) under ice-bath, and stirred at room temperature for 1 hour, LCMS showed the starting material was consumed completely, then p-toluenesulfonic acid (104.22 mg, 604 umol, 3 eq) was added directly to the next step under ice-bath. The theoretical mass of LA-27AA-2-b was 55.16 mg.

[0904] LCMS (ESI): m / z, 274 [M+H] + , Rt = 0.198 min.

[0905] Step 3: Preparation of compound LA-27AA-2-c

[0906] To a solution of LA-27AA-2-b (45.05 mg, 56.53 pmol, 1.1 eq) and LY-22CD-3-d (14.05 mg, 51.39 pmol, 1 eq) in DMF (1 mL) was added DIPEA (26.57 mg, 205.58 pmol, 4 eq) and HATU (23.45 mg, 61.67 pmol, 1.2 eq) under ice-bath and stirred at room temperature for 1 h. LCMS showed the starting material was consumed completely. Purification by high performance liquid chromatography, prep (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 10-40% elution), afforded LA-27AA-2-c (21.30 mg, yield 39.39%) as yellow oil.

[0907] LCMS (ESI): m / z, 1074 [M+Na] + , Rt = 1.241 min.

[0908] Step 4: Preparation of compound LA-27AA-2-d

[0909] To a solution of compound LA-27AA-2-c (21.30 mg, 20.24 pmol, 1 eq) in DMF (2 mL) was added DIPEA (20.93 mg, 161.94 pmol, 8 eq) and NPC (36.95 mg, 121.45 pmol, 6 eq) and stirred at 35 °C for 1.5 h. The solution was concentrated by oil pump. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0-50%, methanol / dichloromethane = 0-8%) to give LA-27AA-2-d (12.30 mg, purity 50%) as yellow oil.

[0910] LCMS (ESI): m / z, 1217 [M+H] + , Rt = 1.937 min.

[0911] Step 5: Preparation of compound LA-27AA-2

[0912] To a solution of compound LA-27AA-2-d (12.30 mg, 50% purity, 5.05 pmol, 1 eq) and eribulin mesylate (4.17 mg, 5.05 pmol, 1 eq) in DMF (1 mL) was added DIPEA (2.61 mg, 20.21 pmol, 4 eq), stirred at room temperature for 1 hour, LCMS showed the starting material was consumed completely. The reaction solution was purified by high performance liquid chromatography preparation (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was eluted from 30-60%), to give white solid LA-27AA-2 (700 pg, yield 7.66%).

[0913] LCMS (ESI): m / z, 1831 [M+Na] + Rt = 2.18 min.

[0914] Example 29: Preparation of compound LA-27AA-3

[0915] Step 1: Preparation of compound LA-27AA-3-c

[0916] To a solution of compound LY-46F-c (200 mg, 87%, 255.62 pmol, 1 eq) in DMF (4 mL) was added 5-amino-1-pentanol (31.64 mg, 32.06 pL, 306.74 pmol, 1.2 eq) and DIPEA (66.07 mg, 84.49 pL, 511.24 pmol, 2 eq), stirred at room temperature for 2 hours, LCMS showed the starting material was consumed completely. The reaction solution was purified by high performance liquid chromatography preparation, to give white solid LA-27AA-3-c (60 mg, yield 48.33%).

[0917] Preparation conditions as follows: preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was YMC-Triart Prep C18 250 x 50 mm x 7 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, the elution ratio of acetonitrile was eluted from 10% to 40%.

[0918] LCMS (ESI): m / z, 445.3 [M+Na] + .

[0919] Step 2: Preparation of compound LA-27AA-3-d

[0920] To a solution of compound LA-27AA-3-c (40 mg, 94.67 pmol, 1 eq) in DMF (2 mL) was added LY-22CD-3-d (75.44 mg, 94.67 pmol, 1 eq) and DIPEA (24.47 mg, 31.29 pL, 189.34 pmol, 2 eq), HATU (54 mg, 142 pmol, 1.5 eq) was added at 0 °C, and the mixture was stirred at room temperature for 3 h. LCMS showed the starting material was consumed completely. The reaction solution was purified by high performance liquid chromatography to give the product LA-27AA-3-d (40 mg, yield 35.17%) as a yellow solid.

[0921] Preparative conditions: preparative chromatography company Shimadzu, model LC-20AP. The column was Phenomenex Luna C18 150 x 25 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution.

[0922] LCMS (ESI): m / z, 1223.7 [M+Na] + .

[0923] Step 3: Preparation of compound LA-27AA-3-e

[0924] To a solution of compound LA-27AA-3-d (20 mg, 16.65 pmol, 1 eq) in DMF (1 mL) was added NPC (25.32 mg, 83.24 pmol, 5 eq) and DIPEA (10.76 mg, 13.76 pL, 83.24 pmol, 5 eq), and the mixture was stirred at room temperature for 6 h. LCMS showed the starting material was consumed completely. The reaction solution was rotary evaporated, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane 1:20 to 1:5) and high performance liquid chromatography (PE:EA = 1:1 and DCM:MeOH = 10:1, R f = 0.4). LA-27AA-3-e (10 mg, yield 43.96%, purity 60%) was obtained as a yellow oil.

[0925] LCMS (ESI): m / z, 1366.5 [M+H] + .

[0926] Step 4: Preparation of compound LA-27AA-3

[0927] To the solution of compound LA-27AA-3-d (10 mg, 60%, 4.39 pmol, 1 eq) in DMF (1 mL) was added eribulin mesylate (3.63 mg, 4.39 pmol, 1 eq) and DIPEA (1.13 mg, 1.45 pL, 8.78 pmol, 2 eq), and stirred at 25 °C for 3 hours. LCMS showed the reaction was completed. The reaction solution was prepared by high performance liquid chromatography to obtain the product (1.81 mg, yield 21.06%, purity 90%) as a white solid.

[0928] High performance liquid chromatography preparation conditions were as follows:

[0929] Preparation chromatography manufacturer Oriendo, model BRIX-2860 (R1, 4, 5, 6). The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 20% to 50% elution.

[0930] LCMS (ESI): m / z, 978.9 [(M+2H) / 2] + .

[0931] Example 30: Preparation of compound LA-27A0

[0932] Step 1: Preparation of compound (9H-fluoren-9-yl)methyl (2-((((5- hydroxypentyl)oxy)methyl)amino)-2-oxoethyl)carbamate

[0933] Methyl (2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)acetamido)acetate (300 mg, 814.38 pmol, 1 eq) was dissolved in THF (6 mL), and 5-((tert- butyldimethylsilyl)oxy)pentan-1-ol (544.49 mg, 2.44 mmol, 3 eq) and TsOH (42.50 mg, 244.31 pmol, 0.3 eq) were added, and the reaction solution was stirred at 25 °C for 1 hour. LCMS showed that the product was generated. The reaction solution was prepared by high phase liquid chromatography to obtain the product LA-27A0-a (200 mg, yield 59.54%) as a white solid.

[0934] High performance liquid chromatography preparation conditions were as follows:

[0935] Preparation chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Synergi Max-RP 200 x 30 mm x 10 pm, and the mobile phase was water (0.225% HCOOH) - acetonitrile, and the elution ratio of acetonitrile was from 20% to 50% elution.

[0936] LCMS (ESI): m / z, 435.3 [M+Na] + .

[0937] Step 2: Preparation of compound (9H-fluoren-9-yl)methyl (2-(((5-(((4- nitrophenoxy)carbonyl)oxy)pentyl)oxymethyl)amino)-2-oxoethyl)carbamate

[0938] (9H-fluoren-9-yl)methyl (2-((((5-hydroxypentyl)oxy)methyl)amino)-2-oxoethyl)carbamate (200 mg, 484.86 pmol, 1 eq) was dissolved in DMF (5 mL), DIPEA (253.19 mg, 1.94 mmol, 4 eq) and NPC (304.13 mg, 969.73 pmol, 2 eq) were added, and the reaction was stirred at 25 °C for 3 h. TLC showed that the starting material disappeared and a new spot with less polarity was generated. The reaction was concentrated, and the residue was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give the product LA-27A0-b (260 mg, yield 92.84%) as yellow oil.

[0939] LCMS (ESI): m / z, 578.2 [M+H] + .

[0940] Step 3: Preparation of compound LA-27A0-c

[0941] Eribulin mesylate was dissolved in DMF (2 mL), DIPEA (20.34 mg, 155.82 pmol, 3 eq) was added, and then (9H-fluoren-9-yl)methyl (2-(((5-(((4-nitrophenoxy)carbonyl)oxy)pentyl)oxymethyl)amino)-2-oxoethyl)carbamate (30 mg, 51.94 pmol, 1 eq) was added, and the reaction was stirred at 25 °C for 1 h. LCMS showed that the product was generated. The reaction was prepared by high performance liquid chromatography to give the product (200 mg, yield 84.36%) as white solid.

[0942] The high performance liquid chromatography preparation conditions were as follows:

[0943] The preparation chromatography manufacturer was Oriendo, model BRIX-2860. The chromatographic column was Synergi Max-RP 200 x 30 mm x 10 pm. The mobile phase was water (0.225% HCOOH)-acetonitrile, and the elution ratio of acetonitrile was eluted from 50% to 80%.

[0944] LCMS (ESI): m / z, 1190.7 [(M+Na] + .

[0945] Step 4: Preparation of compound LA-27A0-d

[0946] To a solution of compound LA-27A0-c (30 mg, 25.68 pmol, 1 eq) in DMF (3 mL), added Et2NH (9.48 mg, 128.38 pmol, 5 eq), the reaction was stirred at 25 °C for 1 h. LCMS showed product was generated. The reaction was concentrated to get the product as yellow oil (24 mg, yield 98.79 %), which was used in the next step directly.

[0947] LCMS (ESI): m / z, 946.6 [(M+H)] + .

[0948] Step 5: Preparation of compound LA-27A0

[0949] To a solution of compound LA-27A0-d (16 mg, 16.91 pmol, 1 eq) in DMF (1 mL), added LY-22CD-3-d (the synthesis of this compound please refer to patent application WO2024175069A1) (17.71 mg, 90 %, 20 pmol, 1 eq) and DIPEA (4.42 mg, 33.82 pmol, 2 eq), added HATU (9.65 mg, 25.37 pmol, 1.5 eq) at 0 °C, the reaction was stirred at 0-10 °C for 1 h. LCMS showed target product. The reaction was prepared by high performance liquid chromatography to get the product as white solid (22.42 mg, yield 76.86 %).

[0950] High performance liquid chromatography preparation conditions as follows:

[0951] Preparation chromatography company Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 150 x 30 mm x 10 pm. The mobile phase was water (0.225 % HCOOH) - acetonitrile, acetonitrile elution ratio from 30 % to 60 % elution.

[0952] LCMS (ESI): m / z, 1724.8 [(M+H] + .

[0953] 1H NMR (400 MHz, DMSO-d6) δ 9.06 (t, J = 5.7 Hz, 1H), 8.86 (d, J = 8.6 Hz, 1H), 8.51 (t, J = 6.6 Hz, 1H), 8.38 (d, J = 8.6 Hz, 1H), 8.12 (t, J = 5.7 Hz, 1H), 7.90 (t, J = 5.6 Hz, 1H), 6.93 (d, J = 5.9 Hz, 1H), 5.05 (s, 1H), 4.99 (s, 1H), 4.82 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.58 - 4.46 (m, 4H), 4.25 (d, J = 10.8 Hz, 1H), 4.21 - 4.12 (m, 1H), 4.10 (s, 3H), 4.01 (t, J = 7.1 Hz, 2H), 3.89 (t, J = 6.6 Hz, 2H), 3.84 - 3.72 (m, 2H), 3.69 (d, J = 5.8 Hz, 2H), 3.65 (s, 3H), 3.63 - 3.59 (m, 3H), 3.53 (d, J = 5.2 Hz, 3H), 3.48 (d, J = 4.9 Hz, 31H), 3.37 (d, J = 5.9 Hz, 3H), 3.25 (s, 4H), 3.18 (q, J = 5.7 Hz, 2H), 2.93 (s, 2H), 2.83 (d, J = 9.5 Hz, 1H), 2.80 - 2.63 (m, 3H), 2.60 - 2.51 (m, 2H), 2.38 (t, J = 6.5 Hz, 2H), 2.36 - 2.29 (m, 4H), 2.25 (d, J = 11.9 Hz, 3H), 2.12 (t, J = 11.8 Hz, 1H), 2.05 - 1.83 (m, 7H), 1.75 - 1.59 (m, 5H), 1.48 (dq, J = 21.0, 6.8 Hz, 8H), 1.30 (q, J = 8.7, 8.1 Hz, 5H), 1.19 - 1.15 (m, 1H), 1.03 (d, J = 6.5 Hz, 3H), 0.99 - 0.92 (m, 1H).

[0954] Example 31: Preparation of compound LA-27AA-20

[0955] Step 1: Preparation of compound LA-27AA-20

[0956] To a solution of compound PE-21EA (11.12 mg, 12.94 pmol, 1 eq) (the synthesis of this compound please refer to the preparation of PE-21EA) and compound LY-22CD-3-d (11.46 mg, 90%, 12.94 pmol, 1 eq) (the synthesis of this compound please refer to patent application WO2024175069A1) in DMF (1 mL) was added DIPEA (6.09 mg, 46.5 pmol, 3.6 eq) and HATU (14.77 mg, 38.83 pmol, 3 eq) at 0 °C. The reaction was stirred at 25 °C for 30 min. LCMS showed product was generated. The reaction was filtered and purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% FA) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution) to give compound LA-27AA-20 (3.25 mg, 5.33% yield, 100% purity) as a white solid after lyophilization.

[0957] LCMS (ESI): m / z, 1638.9 [M+H] + .

[0958] 1H NMR (400 MHz, DMSO-d6) δ 9.05 (t, J = 5.8 Hz, 1H), 8.87 (d, J = 8.6 Hz, 1H), 8.38 (d, J = 8.6 Hz, 1H), 7.88 (s, 1H), 7.77 (s, 1H), 6.90 (s, 1H), 5.05 (s, 1H), 4.99 (s, 1H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.55 (t, J = 4.2 Hz, 1H), 4.51 (d, J = 5.1 Hz, 1H), 4.26 (d, J = 10.6 Hz, 1H), 4.18 (d, J = 10.5 Hz, 1H), 4.12 - 4.08 (m, 3H), 4.02 (s, 1H), 3.89 (t, J = 6.7 Hz, 2H), 3.79 (dd, J = 20.8, 9.5 Hz, 2H), 3.65 (s, 3H), 3.62 (d, J = 6.4 Hz, 1H), 3.58 (d, J = 6.5 Hz, 1H), 3.56 - 3.53 (m, 3H), 3.49 (d, J = 1.5 Hz, 22H), 3.48 - 3.46 (m, 8H), 3.37 (t, J = 5.9 Hz, 2H), 3.26 (d, J = 4.7 Hz, 4H), 3.18 (d, J = 5.8 Hz, 2H), 3.01 (d, J = 6.3 Hz, 2H), 2.94 (s, 2H), 2.84 (d, J = 9.5 Hz, 1H), 2.72 (d, J = 11.6 Hz, 2H), 2.69 - 2.65 (m, 2H), 2.56 (d, J = 6.5 Hz, 1H), 2.35 - 2.29 (m, 6H), 2.29 - 2.24 (m, 4H), 2.20 - 2.12 (m, 2H), 2.04 - 1.95 (m, 2H), 1.92 (s, 4H), 1.73 - 1.62 (m, 5H), 1.51 (q, J = 6.9 Hz, 5H), 1.38 (q, J = 7.3 Hz, 3H), 1.28 (dd, J = 15.0, 8.0 Hz, 4H), 1.17 (d, J = 12.3 Hz, 1H), 1.03 (d, J = 6.4 Hz, 3H), 0.99 - 0.93 (m, 1H).

[0959] Example 32: Preparation of compound LA-27AAVC

[0960] Step 1: Preparation of compound LA-27AAVC-a

[0961] To a solution of compound Fmoc-Val-Cit-OH (3 g, 6.042 mmol, 1 eq) in DMF (10 mL) was added DIEA (2.996 mL, 18.125 mmol, 5 eq), 5-amino-1-pentanol (0.81 g, 7.854 mmol, 1.3 eq) and HATU (2.99 g, 7.854 mmol, 1.3 eq) at room temperature. Then the mixture was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction was concentrated by reduced pressure to get the crude product, the crude product was slurry with ethyl acetate (30 ml) to get LA-27AAVC-a (1.1 g, yield 31.30%) as a yellow solid.

[0962] LCMS (ESI): m / z, 582.2 [M+1] + Rt = 1.336 min.

[0963] Step 2: Preparation of compound LA-27AAVC-b

[0964] To a solution of compound LA-27AAVC-a (1.1 g, 1.891 mmol, 1 eq) in DMF (15 mL) was added DIEA (0.781 mL, 4.727 mmol, 2.5 eq) and 4-nitrophenol methyl ester (1.73 g, 5.673 mmol, 3 eq) at 25 °C. The reaction was stirred at 25 °C for 1 h. LCMS showed the starting material was consumed completely. The reaction was concentrated by reduced pressure to get the crude product, the crude product was slurry with reverse C18 (TFA 0.025%) H2O:ACN = 95%:5%-46%:54%) to get LA-27AAVC-b (770 mg, yield 54.52%) as a white solid.

[0965] LCMS (ESI): m / z, 746.6 [M] + Rt = 1.598 min.

[0966] Step 3: Preparation of compound LA-27AAVC-c

[0967] To compound eribulin mesylate (100 mg, 0.121 mmol, 1 eq) in DMF (3 mL) was added DIEA (47 mg, 0.363 mmol, 3 eq), LEC-27AAVC-b (108.50 mg, 0.145 mmol, 1.2 eq) and HOBt (16.36 mg, 0.121 mmol, 1 eq) at room temperature. Then the mixture was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction solution was purified by high performance liquid chromatography (C18, (0.025% TFA) H2O:ACN = 95%:5% - 18%:82%) to give LA-27AAVC-c (67 mg, yield 41.37%) as a white solid.

[0968] LCMS (ESI): m / z, 1337.4 [M] + , Rt = 1.692 min.

[0969] Step 4: Preparation of compound LA-27AAVC-d

[0970] To a solution of compound LA-27AAVC-c (29 mg, 0.022 mmol, 1 eq) in DMF (1 mL) was added diethylamine (0.022 mL, 0.217 mmol, 10 eq) at 25 °C. The reaction was stirred at 25 °C for 1.5 h. LCMS showed the starting material was consumed completely. The reaction was concentrated in vacuum at low temperature to give crude LA-27AAVC-d (24 mg, crude) as a yellow oil.

[0971] LCMS (ESI): m / z, 1114.1 [M-1] + , Rt = 1.310 min.

[0972] Step 5: Preparation of compound LA-27AAVC

[0973] To a solution of compound LA-27AAVC-d (24 mg, 0.022 mmol, 1 eq) in DMF (2 mL) was added DIEA (0.011 mL, 0.065 mmol, 3 eq), LY-22CD-3-d (the synthesis of this compound can be found in patent application WO2024175069A1) (18.86 mg, 0.024 mmol, 1.1 eq) and HATU (10.64 mg, 0.028 mmol, 1.3 eq) at 0 °C. Then the mixture was stirred at 25 °C for 1 h. LCMS showed the reaction was completed. The reaction solution was purified by high performance liquid chromatography (Bonnasil-BS C18, 20 x 250 mm x 8 pm, H2O (0.1% FA):ACN = 70%:30%-40%:60%) to give LA-27AAVC (18.57 mg, 45.56% yield) as a white solid.

[0974] LCMS (ESI): m / z, 1894.3 [M] + Rt = 1.402 min.

[0975] 1H NMR (400 MHz, DMSO) δ 9.05 (t, J = 5.5 Hz, 1H), 8.89 - 8.85 (m, 1H), 8.38 (d, J = 8.6 Hz, 1H), 7.87 (dd, J = 13.5, 6.5 Hz, 2H), 7.78 (t, J = 5.2 Hz, 1H), 6.92 (t, J = 5.6 Hz, 1H), 5.96 (d, J = 14.9 Hz, 1H), 5.05 (s, 1H), 5.00 (s, 1H), 4.83 (s, 1H), 4.75 (s, 1H), 4.63 (s, 1H), 4.55 (t, J = 4.0 Hz, 1H), 4.26 (d, J = 10.8 Hz, 1H), 4.21 - 3.97 (m, 7H), 3.89 (t, J = 6.4 Hz, 2H), 3.79 (dd, J = 21.5, 9.5 Hz, 3H), 3.70 - 3.61 (m, 8H), 3.55 - 3.49 (m, 38H), 3.37 (d, J = 5.8 Hz, 5H), 3.25 (s, 3H), 3.23 - 3.15 (m, 3H), 3.02 (d, J = 6.0 Hz, 2H), 2.94 (d, J = 5.4 Hz, 4H), 2.84 (d, J = 9.5 Hz, 1H), 2.79 - 2.66 (m, 2H), 2.66 - 2.53 (m, 2H), 2.46 (dd, J = 13.6, 6.5 Hz, 1H), 2.35 (dt, J = 19.9, 6.4 Hz, 4H), 2.28 - 2.18 (m, 3H), 2.17 - 2.07 (m, 1H), 2.06 - 1.85 (m, 7H), 1.66 (dt, J = 22.6, 11.2 Hz, 5H), 1.48 (dd, J = 14.6, 8.1 Hz, 5H), 1.41 - 1.16 (m, 12H), 1.02 (t, J = 10.0 Hz, 3H), 0.82 (t, J = 7.6 Hz, 5H).

[0976] Example 33: Preparation of compound LA-27AAVCP

[0977] Step 1: Preparation of compound LA-27AAVC-c

[0978] To a solution of compound eribulin mesylate (commercially available) (50.0 mg, 0.061 mmol, 1.0 eq) and DIEA (23.47 mg, 0.182 mmol, 3.0 eq) in DMF (1.0 mL) was added LA-27AAVC-b (see example of LA-27AAVC) (45.56 mg, 0.061 mmol, 1.0 eq) and HOBt (8.18 mg, 0.061 mmol, 1.0 eq) at room temperature and stirred at room temperature for 1 h, LCMS showed the reaction was complete. The reaction was filtered, the filtrate was purified by C18 reverse phase column chromatography, mobile phase water (0.25% TFA) - acetonitrile, elution ratio of acetonitrile from 0% to 60%, to give the product LA-27AAVC-c as a white solid (61.0 mg, yield 75.35%).

[0979] LCMS (ESI): m / z, 1337.1 [(M+H)] + .

[0980] Step 2: Preparation of compound LA-27AAVC-d

[0981] To a solution of compound LA-27AAVC-c (60 mg, 0.045 mmol, 1.0 eq) in DMF (2.0 mL) was added DEA (32.81 mg, 0.449 mmol, 10.0 eq) at room temperature and stirred at room temperature for 0.5 h, LCMS showed the reaction was complete. The reaction was concentrated under reduced pressure to give the crude product LA-27AAVC-d as a brown oil (60 mg, crude).

[0982] LCMS (ESI): m / z, 1115.0 [(M+H)] + .

[0983] Step 3: Preparation of compound LA-27AAVCP

[0984] To a solution of compound LA-27AAVC-d (60 mg, crude, 1.0 eq) and LA-27AAVC-d-1 (the synthesis of this compound please refer to patent application WO2024175069A1) (37.21 mg, 0.054 mmol, 1.2 eq) in DMF (2.0 mL) was added HATU (25.67 mg, 0.067 mmol, 1.5 eq) at room temperature and stirred at room temperature for 1 h. LCMS showed the reaction was complete. The reaction solution was filtered and the filtrate was purified by high performance liquid chromatography preparation (model JB-C235-04, the chromatographic column was Bonnasil-BS C18, 20 x 250 mm x 8 pm, mobile phase water (0.1% TFA) - acetonitrile, acetonitrile elution ratio from 20% to 60% elution) to give the product LA-27AAVCP (55.9 mg, yield 69.70%) as a white solid.

[0985] LCMS (ESI): m / z, 895.6 [(M / 2+H)] + .

[0986] 1 HNMR (400 MHz, DMSO) δ 9.12 (s, 2H), 8.00 - 7.72 (m, 5H), 6.92 (s, 1H), 5.98 (s, 2H), 5.03 (d, J = 23.7 Hz, 2H), 4.80 (d, J = 30.9 Hz, 2H), 4.64 (s, 1H), 4.56 (t, J = 4.1 Hz, 1H), 4.27 (d, J = 10.2 Hz, 1H), 4.21 - 4.14 (m, 3H), 4.11 (s, 3H), 4.02 (s, 1H), 3.89 (t, J = 6.6 Hz, 2H), 3.79 (dd, J = 22.6, 10.1 Hz, 3H), 3.68 (s, 2H), 3.50 (d, J = 2.5 Hz, 28H), 3.26 (s, 3H), 3.21 (dd, J = 11.6, 5.8 Hz, 4H), 3.06 - 2.82 (m, 9H), 2.80 - 2.53 (m, 6H), 2.41 - 1.59 (m, 28H), 1.57 - 1.11 (m, 19H), 1.04 (d, J = 6.4 Hz, 3H), 0.86 - 0.79 (m, 6H).

[0987] Example 34: Preparation of compound LA-27BCN

[0988] Step 1: Preparation of compound LA-27BCN-a

[0989] To a solution of N-fluorenylmethoxycarbonyl-ethanediol-carboxylic acid (500 mg, 1.41 mmol, 1 eq) in dichloromethane (8 mL) was added N-hydroxysuccinimide (194.30 mg, 1.69 mmol, 1.2 eq) and N,N'-dicyclohexylcarbodiimide (580.56 mg, 716.74 μL, 2.81 mmol, 2 eq) under ice-bath and stirred at room temperature for 2 h, LCMS showed the starting material was consumed completely. After filtration, the residue was concentrated to give crude LA-27BCN-a (1.19 g, crude without yield) as colorless oil.

[0990] LCMS (ESI): m / z, 475 [M+Na] + , Rt = 0.944 min.

[0991] Step 2: Preparation of compound LA-27BCN-b

[0992] To a solution of LA-27BCN-a (600 mg, 53%, 702.82 μmol, 1 eq) in DMF (5 mL) was added amino-PEG(8)-acid (351.88 mg, 773.11 μmol, 1.1 eq) and DIPEA (232.31 μL, 1.41 mmol, 2 eq) at 0 °C and stirred at room temperature for 1 h, LCMS showed the starting material was consumed completely, the residue was purified by silica gel column chromatography (C18, ISCO, R-330 g SepaFlash Silica Flash Column, Eluent of 5-25% water / CH3CN, 100 mL / min) to give LA-27BCN-b (456.80 mg, yield 83.45%) as yellow oil.

[0993] LCMS (ESI): m / z, 779 [M+H] + , Rt = 1.833 min.

[0994] Step 3: Preparation of compound LA-27BCN-c

[0995] To a solution of compound LA-27BCN-b (456.80 mg, 586.48 μmol, 1 eq) in DMF (8 mL) was added Et2NH (305.13 μL, 2.93 mmol, 5 eq) and stirred at room temperature for 1 h, LCMS showed the starting material was consumed completely, the oil pump was directly used for the next step. The theoretical mass of LA-27BCN-c was 326.46 mg, crude without yield.

[0996] LCMS (ESI): m / z, 557 [M+H] +Rt = 0.600.

[0997] Step 4: Preparation of compound LA-27BCN-d

[0998] To a solution of compound LY-29C-b (101.57 mg, 322.11 μmol, 1.1 eq, the synthesis of this compound please refer to the preparation of LY-29C-c) in DMF (3 mL) was added compound LA-27BCN-c (163 mg, 292.82 μmol, 1 eq) and DIPEA (232.31 μL, 1.41 mmol, 2 eq) at 0 °C and stirred at room temperature for 1 h, LCMS showed the starting material was consumed completely (2 batches, same scale). The residue was prepared by high performance liquid chromatography (preparative chromatography manufacturer Shimadzu, model LC-20AP. The chromatographic column was Welch Xtimate C18 250 x 30 mm x 10 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 20% to 50% elution), to get yellow oil LA-27BCN-d (232.2 mg, yield 30%, purity 70%).

[0999] LCMS (ESI): m / z, 733 [M+H] + Rt = 1.511.

[1000] Step 5: Preparation of compound LA-27BCN

[1001] To a solution of compound LA-27BCN-d (8.99 mg, 12.27 μmol, 1.3 eq), compound LA-27-d (8.99 mg, 12.27 μmol, 1.3 eq, the synthesis of this compound please refer to the example of LA-27) in DMF (3 mL) was added DIPEA (6.16 mg, 99%, 7.88 μL, 47.20 μmol, 5 eq) and HATU (4.31 mg, 11.33 μmol, 1.2 eq) at 0 °C, and stirred at room temperature for 40 min, LCMS showed the starting material was consumed completely. The reaction was prepared by high performance liquid chromatography (preparative chromatography manufacturer Oriendo, model BRIX-2860. The chromatographic column was Welch Xtimate C18 150 x 21.2 mm x 5 μm. The mobile phase was water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to get white solid compound LA-27BCN (14.75 mg, yield 88.07%).

[1002] LCMS (ESI): m / z, 1797.1 [M+H] + Rt = 2.459 min.

[1003] 1 H NMR (400 MHz, DMSO-d6) δ 8.53 (t, J = 6.6 Hz, 1H), 8.04 (d, J = 7.2 Hz, 1H), 7.92 - 7.81 (m, 2H), 7.04 (s, 1H), 6.92 (s, 1H), 5.02 (d, J = 22.6 Hz, 2H), 4.79 (d, J = 31.5 Hz, 2H), 4.63 (p, J = 2.3 Hz, 1H), 4.58 - 4.43 (m, 4H), 4.30 - 4.13 (m, 4H), 4.12 - 3.96 (m, 5H), 3.89 (t, J = 6.6 Hz, 2H), 3.84 - 3.72 (m, 2H), 3.68 (s, 1H), 3.57 (dt, J = 8.5, 6.7 Hz, 4H), 3.50 (d, J = 3.9 Hz, 24H), 3.39 (t, J = 5.9 Hz, 2H), 3.36 (d, J = 6.0 Hz, 22H), 3.25 (s, 4H), 3.19 (q, J = 5.8 Hz, 2H), 3.09 (q, J = 5.9 Hz, 2H), 2.98 - 2.90 (m, 2H), 2.87 - 2.65 (m, 4H), 2.62 - 2.54 (m, 1H), 2.45 - 2.10 (m, 13H), 2.02 - 1.87 (m, 6H), 1.75 - 1.59 (m, 5H), 1.57 - 1.42 (m, 7H), 1.38 - 1.25 (m, 5H), 1.21 (d, J = 7.1 Hz, 3H), 1.03 (d, J = 6.4 Hz, 3H), 0.83 (dd, J = 13.4, 6.8 Hz, 6H).

[1004] Example 35: Preparation of compound LA-27C

[1005] Step 1: Preparation of compound LA-27C-a

[1006] To a solution of compound LA-27-c (the synthesis of this compound please refer to Example of LA-27) (10.60 mg, 8.27 μmol, 1 eq) in DCM (3 mL) was added Dess-Martin Oxidizer (21.48 mg, 98%, 15.77 μL, 49.63 μmol, 6 eq) at 0 °C, after addition, the reaction was stirred at 25 °C for 16 h, LCMS showed the reaction was completed, the reaction solution was combined with another small batch of reaction solution, poured into a mixture of ice water / saturated sodium bicarbonate / sodium sulfite (1V / 1V / 1V, total volume 60 mL), stirred for 5 min, extracted with DCM for 3 times, 40 mL each time, the organic phase was combined, washed with a mixture of saturated sodium bicarbonate / sodium sulfite (1V / 1V, total volume 30 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated to give compound LA-27C-a (14 mg, crude, 1.28 batches, another 0.28 times scale of small test product and this product were combined) as a yellow solid.

[1007] LCMS (ESI): m / z, 1279.9 [M+H] + Rt = 3.257 min.

[1008] Step 2: Preparation of compound LA-27C-b

[1009] To a solution of compound LA-27C-a (14 mg, 10.94 μmol, 1 eq) in DMF (1 mL) was added diethylamine (4 mg, 54.71 μmol, 5 eq) at 0 °C, after addition, the reaction was stirred at 25 °C for 1 h, LCMS showed the reaction was completed, the reaction solution was directly rotary evaporated to give compound LA-27C-b (12 mg, crude) as a yellow solid.

[1010] LCMS (ESI): m / z, 1057.9 [M+H] + .

[1011] Step 3: Preparation of compound LA-27C

[1012] To a solution of compound LA-27C-b (12 mg, 11.35 pmol, 1 eq) and LY-22CD-3-d (18.09 mg, 22.70 pmol, 2 eq) in DMF (2 mL) was added DIPEA (11.73 mg, 90.80 pmol, 8 eq) and HATU (8.63 mg, 22.70 pmol, 2 eq) at 0 °C. After addition, the mixture was stirred at 25 °C for 2 h. LCMS showed the reaction was completed. The reaction solution was purified by high performance liquid chromatography preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 30 mm x 10 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution), and then lyophilized to give compound LA-27C (5.12 mg, 24.57% yield, 98% purity) as a white solid.

[1013] LCMS (ESI): m / z, 1859.1 [M+Na] + Rt = 2.329 min.

[1014] 1H NMR (400 MHz, DMSO-d6) δ 9.06 (t, J = 5.7 Hz, 1H), 8.87 (d, J = 8.6 Hz, 1H), 8.54 (t, J = 6.5 Hz, 1H), 8.38 (d, J = 8.6 Hz, 1H), 8.04 (d, J = 7.2 Hz, 1H), 7.92 - 7.83 (m, 2H), 7.28 (t, J = 5.9 Hz, 1H), 5.07 - 4.99 (m, 2H), 4.82 (s, 1H), 4.75 (s, 1H), 4.63 (dt, J = 4.5, 2.5 Hz, 1H), 4.59 - 4.37 (m, 4H), 4.30 - 4.15 (m, 4H), 4.12 - 3.99 (m, 5H), 3.92 (t, J = 6.6 Hz, 2H), 3.86 - 3.76 (m, 3H), 3.71 (s, 1H), 3.66 - 3.52 (m, 8H), 3.51 - 3.43 (m, 24H), 3.39 - 3.34 (m, 24H), 3.23 (s, 3H), 3.18 (q, J = 5.8 Hz, 3H), 2.87 - 2.71 (m, 4H), 2.67 (td, J = 4.0, 2.0 Hz, 2H), 2.57 (dd, J = 16.5, 9.7 Hz, 2H), 2.45 - 2.36 (m, 2H), 2.33 - 2.11 (m, 8H), 2.03 - 1.86 (m, 7H), 1.66 (p, J = 13.6, 12.7 Hz, 5H), 1.48 (dq, J = 21.9, 7.5 Hz, 6H), 1.37 - 1.17 (m, 9H), 1.00 (dd, J = 22.7, 9.1 Hz, 4H), 0.82 (dd, J = 13.5, 6.8 Hz, 6H).

[1015] Example 36: Preparation of compound LA-27CBCN

[1016] To a solution of compound LA-27C-b (the synthesis of this compound please refer to example of LA-27C, 10.50 mg, 9.93 pmol, 1 eq), compound LA-27BCN-d (the synthesis of this compound please refer to example of LA-27BCN, 14.56 mg, 60%, 11.92 pmol, 1.2 eq) in DMF (8 mL) was added DIPEA (5.19 mg, 39.72 pmol, 4 eq) and HATU (4.53 mg, 11.92 pmol, 1.2 eq) under ice-bath. The reaction was stirred at 25 °C for 1 h. The reaction mixture was purified by high performance liquid chromatography-preparation (Oriendo, BRIX-2860. Column: Welch Xtimate C18 150 x 21.2 mm x 5 pm. Mobile phase: water (0.225% HCOOH) - acetonitrile, acetonitrile elution ratio from 30% to 60% elution) to give compound LA-27CBCN (7.57 mg, 43.01% yield, 96% purity) as white solid.

[1017] LCMS (ESI): m / z, 1795.1 [M+Na] + Rt = 2.603 min.

[1018] 1H NMR (400 MHz, DMSO-d6) δ 8.53 (t, J = 7.0 Hz, 1H), 8.04 (d, J = 7.3 Hz, 1H), 7.92 - 7.82 (m, 2H), 7.28 (t, J = 5.9 Hz, 1H), 7.03 (d, J = 5.6 Hz, 1H), 5.04 (d, J = 11.0 Hz, 2H), 4.82 (s, 1H), 4.75 (s, 1H), 4.63 (q, J = 2.9 Hz, 1H), 4.58 - 4.44 (m, 3H), 4.29 - 4.07 (m, 7H), 4.03 (dd, J = 9.0, 4.7 Hz, 4H), 3.92 (t, J = 6.6 Hz, 2H), 3.87 - 3.64 (m, 5H), 3.63 - 3.54 (m, 4H), 3.54 - 3.43 (m, 24H), 3.39 (t, J = 5.9 Hz, 2H), 3.34 (d, J = 6.4 Hz, 6H), 3.25 - 3.13 (m, 5H), 3.09 (q, J = 6.0 Hz, 2H), 2.87 - 2.53 (m, 7H), 2.48 - 2.25 (m, 11H), 2.17 (dd, J = 27.8, 11.8 Hz, 8H), 1.94 (d, J = 13.0 Hz, 7H), 1.68 (q, J = 12.3 Hz, 4H), 1.50 (dt, J = 24.5, 7.1 Hz, 8H), 1.29 (dq, J = 16.3, 8.6 Hz, 6H), 1.21 (d, J = 7.1 Hz, 5H), 1.00 (dd, J = 22.6, 9.2 Hz, 4H), 0.83 (dd, J = 13.3, 6.8 Hz, 7H).

[1019] Example 37: Preparation of compound LA-27C-2

[1020] Step 1: Prepa...

Claims

1. A derivative of eribulin, which is a compound of Formula (I), Formula (II), or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: A1in formula (I) is selected from A2in formula (II) is selected from at the * and ** places each independently is a single, double or oxygen bond =0, and at least one is not a double bond; R1is selected from H, alkyl, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl or cyano-substituted alkyl; X1is selected from -CO-, -CH2-, -S(O)- or -S(O)2-; X2is selected from a direct bond, -O- or -NR1-; X3is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH or Y1is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH or R2, R3are each independently selected from H, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl, substituted or unsubstituted carbocyclyl-substituted alkyl, substituted or unsubstituted heterocyclyl-substituted alkyl, substituted or unsubstituted carbocyclyl or substituted or unsubstituted heterocyclyl; said substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl means optionally substituted with one or more substituents selected from oxo, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl; R4is selected from substituted or unsubstituted carbocyclyl or substituted or unsubstituted heterocyclyl; said substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl means optionally substituted with one or more substituents selected from oxo, alkyl, hydroxy, alkyloxy, amino, halo, cyano, hydroxy-substituted alkyl, alkyloxy-substituted alkyl, amino-substituted alkyl, haloalkyl, cyano-substituted alkyl; the ring is a substituted or unsubstituted heterocyclyl, said heterocyclyl containing at least one -NH- in the ring, said substitution means substitution with one or more substituents selected from oxo, alkyl, alkyloxy, halo, cyano, alkyloxy-substituted alkyl, haloalkyl, cyano-substituted alkyl; the ring is a heterocyclyl, said heterocyclyl containing at least one -NH- in the ring and / or said heterocyclyl is substituted with amino, hydroxy, amino-substituted alkyl or hydroxy-substituted alkyl, said heterocyclyl being optionally substituted with one or more substituents selected from oxo, alkyl, alkyloxy, halo, cyano, alkyloxy-substituted alkyl, haloalkyl, cyano-substituted alkyl; m1is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; m2is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

2. The derivative of eribulin according to claim 1, characterized in that, said alkyl is C 1-12 linear or branched alkyl; said carbocyclic group is a saturated, partially unsaturated or fully unsaturated carbocyclic group having 3 to 20 carbon atoms; said heterocyclic group is a saturated, partially unsaturated or fully unsaturated heterocyclic group having 3 to 20 atoms, containing 1, 2, 3, 4 heteroatoms selected from N, O, S.

3. The derivative of eribulin according to claim 1 or 2, characterized in that, R1is selected from H, C 1-6 alkyl, hydroxy-substituted C 1-6 alkyl, preferably selected from H, methyl, ethyl, n-propyl, i-propyl, hydroxymethyl, hydroxyethyl, hydroxy-n-propyl, hydroxy-i-propyl.

4. The derivative of eribulin according to any one of claims 1 to 3, wherein, X1is selected from -CO-, -CH2-.

5. The derivative of eribulin according to any one of claims 1 to 4, wherein R2, R3are each independently selected from the group consisting of H, C 1-6 alkyl, hydroxy, C 1-6 alkyloxy, C 1-6 alkyl, C 1-6 alkyloxy-substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl-substituted C 1-6 alkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1 or 2 heteroatoms N or O-substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1 or 2 heteroatoms N or O; the substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl containing 1 or 2 heteroatoms N or O means optionally substituted by oxo, C 1-6 alkyl, hydroxy, C 1-6 alkyloxy, C 1-6 alkyl, C 1-6 alkyloxy-substituted C 1-6 one or more substituents of the alkyl group; R2, R3are each independently selected from the group consisting of H, C 1-6 alkyl, hydroxy, hydroxy-substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl-substituted C 1-6 alkyl, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted C 3-6 cycloalkyl, substituted or unsubstituted C 1-6 alkyl, hydroxy, hydroxy-substituted C 1-6 one or more substituents selected from the group consisting of oxo, C 6. The derivative of eribulin according to any one of claims 1 to 5, wherein, R4 is selected from substituted or unsubstituted C 3-12 monocyclic, annelated, fused, spiro or bridged ring cycloalkyl, substituted or unsubstituted 3-12 membered monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, substituted or unsubstituted C 6-10 aryl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms N or O; the substitution means substituted with one or more substituents selected from oxo, C 1-6 alkyl, hydroxy, C 1-6 alkyloxy, hydroxy substituted C 1-6 alkyl, C 1-6 alkyloxy substituted C 1-6 one or more substituents of alkyl; R4 is selected from substituted or unsubstituted C 4-12 monocyclic, annelated, fused, spiro or bridged ring cycloalkyl, substituted or unsubstituted 4-12 membered monocyclic, annelated, fused, spiro or bridged ring heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, substituted or unsubstituted phenyl, substituted or unsubstituted 5-10 membered heteroaryl containing 1, 2 or 3 heteroatoms N or O, the substitution meaning substituted with one or more substituents selected from oxo, C 1-6 alkyl.

7. The derivative of eribulin according to any one of claims 1 to 6, wherein, Ring is a substituted or unsubstituted 3-12 membered monocyclic, annelated, fused, spiro, or bridged heterocycloalkyl containing 1, 2, or 3 heteroatoms N or O, containing at least one -NH- on the ring of said heterocycloalkyl, said substitution being substitution with one or more substituents selected from oxo, C 1-6 alkyl, C 1-6 alkyloxy, C 1-6 alkyloxy-substituted C 1-6 alkyl.

8. The derivative of eribulin according to any one of claims 1 to 4, wherein, X3is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH or said ring is a substituted or unsubstituted 4-12 membered monocyclic, fused, bicyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said substitution means substituted with one or more substituents selected from oxo, C 1-6 alkyl; ml is selected from 1, 2, 3, 4, 5, 6; m2 is selected from 0, 1, 2, 3, 4, 5 or 6; R2, R3are each independently selected from H, C 1-6 alkyl, hydroxy, hydroxy substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, said substituted or unsubstituted C 3-6 cycloalkyl means optionally substituted with one or more substituents selected from oxo, C 1-6 alkyl; R4is selected from C 4-12 monocyclic, fused, bicyclic, spirocyclic or bridged cyclic cycloalkyl, 4-12 membered monocyclic, fused, bicyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, phenyl.

9. The derivative of eribulin according to claim 8, wherein, X3is selected from the following groups: Preferably, X3is selected from the following groups:

10. The derivative of eribulin according to any one of claims 1 to 4, wherein, Y1is selected from the group consisting of -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, or said ring is a substituted or unsubstituted 4-12 membered monocyclic, fused, bicyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said substitution means substituted with one or more substituents selected from oxo, C 1-6 alkyl; m1 is selected from 1, 2, 3, 4, 5, 6; m2 is selected from 0, 1, 2, 3, 4, 5 or 6; R2, R3 are each independently selected from H, C 1-6 alkyl, hydroxy, hydroxy substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl substituted C 1-6 alkyl, substituted or unsubstituted C 3-6 cycloalkyl, said substituted or unsubstituted C 3-6 cycloalkyl means optionally substituted with one or more substituents selected from oxo, C 1-6 alkyl; R4 is selected from C 4-12 monocyclic, fused, bicyclic, spirocyclic or bridged cyclic cycloalkyl, 4-12 membered monocyclic, fused, bicyclic, spirocyclic or bridged cyclic heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, phenyl.

11. The derivative of eribulin according to claim 10, wherein, Y1is selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH; m1 is selected from 1, 2, 3, 4, 5, 6; R2, R3are each independently selected from H, C 1-6 alkyl, hydroxy, hydroxy-substituted C 1-6 alkyl; preferably, Y1is selected from 12. The derivative of eribulin according to claim 1, wherein, Ring is a 3-12 membered monocyclic, annelated, fused, spiro, or bridged heterocycloalkyl containing 1, 2, or 3 heteroatoms N or O, containing at least one -NH- on the ring of said heterocycloalkyl and / or said heterocycloalkyl being substituted by amino, hydroxy, amino-substituted C 1-6 alkyl or hydroxy-substituted C 1-6 alkyl, said heterocycloalkyl being optionally substituted by one or more substituents selected from the group consisting of oxo, C 1-6 alkyl, C 1-6 alkyloxy, C 1-6 alkyloxy-substituted C 1-6 alkyl.

13. The derivative of eribulin according to claim 12, wherein, ring is a 4-12 membered monocyclic, annelated, fused, spiro or bridged heterocycloalkyl containing 1, 2 or 3 heteroatoms N or O, said heterocycloalkyl containing at least one -NH- in the ring and / or said heterocycloalkyl being substituted by amino, hydroxy, amino-substituted C 1-6 alkyl or hydroxy-substituted C 1-6 alkyl-substituted; preferably, the ring is selected from the following groups:

14. The derivative of eribulin according to claim 1, wherein, A1, A2are selected from the following groups: A2in formula (II) is also selected from the group consisting of Preferably, A1, A2are selected from the following groups:

15. The derivative of eribulin according to claim 1, wherein, The compound of formula (I) is one of the following: In particular, the compound of formula (II) is one of the following:

16. An eribulin derivative which is a compound of Formula (I’), Formula (II’), or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Q is a linker group that can be attached to an antibody or antigen binding fragment; L is a linker group connecting Q to A1' or Q to A2'; A1' is a group of A1 attached to L after removal of the H of -NH2, -OH, -NH- in a chain or -NH- in a ring; A2' is a group of A2 attached to L after removal of the H of -NH2, -OH, -NH- in a chain or -NH- in a ring; A1, A2, *, ** are as defined in any one of claims 1-15 in the definitions of formula (I), formula (II).

17. The derivative of eribulin according to claim 16, wherein when X3, Y1are independently selected from -(CR2R3) m1 -NH2, -(CR2R3) m1 -OH, -R4-(CR2R3) m2 -NH2, -R4-(CR2R3) m2 -OH, A1' is a group which A1is linked to L after removal of H of terminal -NH2or terminal -OH of X3, Y1, and A2' is a group which A2is linked to L after removal of H of terminal -NH2or terminal -OH of X3, Y1. when X3, Y1are each independently selected from at the time, A1' is A1 by removal of the group attached to L after the H of the ring -NH- is removed a group of -NH2, -OH in a ring attached to L after removal of the H of -NH2, -OH in a ring; when A1, A2are each independently selected from at the time, A1' is A1 by removal of H or the ring the H of -NH2, -OH on the ring substituent, the group to which L is attached after the removal of H or the ring a group of -NH2, -OH in a ring attached to L after removal of the H of -NH2, -OH in a ring.

18. The derivative of eribulin according to claim 16, wherein, R1is H, A1' is a group of A1 which is linked to L after removal of R1of -NR1-; A2' is a group of A2 which is linked to L after removal of R1of -NR1-.

19. The derivative of eribulin according to any one of claims 16-18, wherein, L is -L4-L3-L2-L1-, wherein L1is linked to A1' or A2', and L4is linked to Q; L1is selected from a direct bond, wherein for the linking site to A1' or A2', is a linking site to L2; L2is a direct bond, one amino acid residue, or a short peptide chain formed by two or more amino acid residues selected from natural amino acid residues or unnatural amino acid residues, the N-terminal of which is connected to L3, the C-terminal of which is connected to L1, and L2optionally comprises one or more structures selected from the group consisting of formula (III-1) to formula (III-9): s is selected from 1, 2, 3, 4, 5, 6, 7, 8, t is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, R5is selected from H or C 1-8 alkyl; L3is selected from a direct bond, or L3 is selected from r1 is selected from 0, 1, 2, 3, 4, 5, 6, r2 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r3 is selected from 1, 2, 3, 4, 5, 6, r4 is selected from 1, 2, 3, 4, 5, 6, r5 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r6 is selected from 0, 1, 2, 3, 4, 5, 6, r7 is selected from 0, 1, 2, 3, 4, 5, 6, r8 is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, r9 is selected from 1, 2, 3, 4, 5, 6, r10 is selected from 1, 2, 3, 4, 5, 6, W is selected from O or NR6, R6 is selected from H or C 1-8 alkyl, wherein represents the linking site to L2, 2 occurrences in the same L3 group represents that both sites are linked to L2; L4is selected from a direct bond, -CO-, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2) p4 -O-(CH2) p5 -C(O)-, -O-(CH2) p1 -CO-, -O-(CH2CH2O) p2 -(CH2) p3 -CO-, -O-(CH2) p4 -O-(CH2) p5 -C(O)-, p1, p2, p3, p4, p5 are each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, R7is selected from H or C 1-8 alkyl, wherein -CO- is attached to L3.

20. The derivative of eribulin according to claim 19, wherein, A1' is a group of A1 attached to L after removal of the H of -NH2, -NH- in a chain or -NH- in a ring, or A2' is a group of A2 attached to L after removal of the H of -NH2, -NH- in a chain or -NH- in a ring, L1 is selected from a direct bond, A1' is a group of A1 after removal of the H of -OH followed by attachment to L, or A2' is a group of A2 after removal of the H of -OH followed by attachment to L, L1 is selected from 21. The derivative of eribulin according to claim 19 or 20, wherein, L1is selected from a direct bond, 22. The derivative of eribulin according to any one of claims 19-21, wherein, L2is an amino acid residue, or a short peptide chain formed by 2-8 amino acid residues, preferably the amino acid residue is an amino acid residue formed by an amino acid selected from the group consisting of phenylalanine, alanine, glycine, valine, citrulline, leucine, isoleucine, tryptophan, tyrosine, histidine, lysine, serine, threonine, cysteine, glutamic acid, glutamine, aspartic acid, asparagine, methionine, arginine; more preferably the amino acid residue is an amino acid residue formed by an amino acid selected from the group consisting of phenylalanine, alanine, glycine, valine, citrulline.

23. The derivative of eribulin according to claim 22, wherein, L2optionally comprises one or more structures selected from the group consisting of Formula (III-1), Formula (III-9); preferably, optionally comprises one or more structures selected from the group consisting of one or more of the following structures:

24. The derivative of eribulin according to claim 22 or 23, wherein, L2 is selected from NH Gly CO - or from the following short peptide chains: NH Gly-Gly-Phe-Gly CO -, NH Val-Cit CO -, NH Val-Ala CO -, NH Gly-Gly-Val-Ala CO -, NH Gly-Gly-Val-Cit CO -, NH Lys-Gly-Val-Ala CO -, NH Lys-Gly-Val-Cit CO -, NH Gly-Val-Cit CO -.

25. The derivative of eribulin according to claim 24, wherein L2is selected from the following groups: Preferably, L2is selected from the following groups:

26. The derivative of eribulin according to any one of claims 19 to 25, wherein, L3is selected from a direct bond, or L3is selected from r1, r6, r7are selected from 0, r2is selected from 4, 5, 6, 7, 8, r3is selected from 1, 2, 3, 4, r4is selected from 1, 2, 3, 4, r5is selected from 4, 5, 6, 7, 8, r8is selected from 1, 2, 3, 4, r9is selected from 1, 2, 3, 4, r10is selected from 1, 2, 3, 4, W is selected from NH; Preferably, L3is selected from a direct bond, r1, r7 are selected from 0, r2 is selected from 4, 5, 6, 7, 8, r3 is selected from 2, r8 is selected from 2, 3, 4, r9 is selected from 2, r10 is selected from 2.

27. The derivative of eribulin according to any one of claims 19 to 26, wherein, L4is selected from a direct bond, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2) p4 -O-(CH2) p5 -C(O)-, pi is selected from 1, 2, 3, 4, 5, 6, p2, p3, p4, p5 are each independently selected from 1, 2, 3, 4, R7is selected from H or C 1-6 alkyl; preferably, L4is selected from a direct bond, -(CH2) p1 -CO-, -(CH2CH2O) p2 -(CH2) p3 -CO-, -(CH2) p4 -O-(CH2) p5 -C(O)-, -NR7-(CH2) p1 -CO-, -NR7-(CH2CH2O) p2 -(CH2) p3 -CO-, -NR7-(CH2) p4 -O-(CH2) p5 -C(O)-, pi is selected from 3, 4, 5, 6, p2, p3, p4, p5 are each independently selected from 2, 3, R7is selected from H.

28. The derivative of eribulin according to any one of claims 16 to 27, wherein, Q is selected from a group comprising a maleimide linker, a group comprising a methanesulfonyl pyrimidine linker, a group comprising a methanesulfonyl pyridothiazole linker, a group comprising a methanesulfonyl pyrimidothiazole linker, a group comprising a cycloalkene alkyne linker, or a group comprising an oxime bond linker.

29. The derivative of eribulin according to claim 28, wherein, Q is selected from 30. The derivative of eribulin according to claim 29, wherein Q-L4 is selected from Preferably, Q-L4 is selected from 31. The derivative of eribulin according to claim 16, wherein Formula (I') is one of the following: Formula (II') is one of the following:

32. An antibody drug conjugate, which is a compound of Formula (I”), Formula (II”), or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: Ab is an antibody or an antigen binding fragment; L5 is a direct bond or a linker linking Ab to Q'; Q' is a group of Q which is linked to Ab through L5; n is an integer or a decimal number between 0 and 20; preferably n is an integer or a decimal number between 0 and 8; Q, L, A1', A2', *, ** are as defined in any one of claims 16-31 in formula (I'), formula (II').

33. The antibody drug conjugate of claim 32, wherein, L5is a direct bond or represents a linking site for attachment to Ab, represents Z3attached to Q'; Z3is a click probe or a thiol or a precursor thereof that is capable of attaching to Q' after reaction; Z1and Z2are each independently selected from -C(O)NH-, -C(O)O-, -C(O)-, -OC(O)-, -NHC(O)-, -NH-, -O-, -OC(O)NH-, or -NHC(O)O-; q1 is 0, 1, 2, 3, 4, 5, 6, 7, 8; q2 is 1, 2, 3, 4, 5, 6, 7, 8; q3 is 1, 2, 3, 4, 5, 6, 7, 8; q4 is 1, 2, 3, 4, 5, 6, 7, 8.

34. The antibody drug conjugate of claim 33, wherein, Z3is a click probe or a thiol group or a precursor thereof which is capable of being linked to Q' after a metal-free click reaction, preferably Z3is an azido group.

35. The antibody drug conjugate of any one of claims 32-34, wherein, L5is a direct bond or Preferably, L5is a direct bond or 36. The antibody drug conjugate of any one of claims 32-35, wherein, Z1and Z2are each independently selected from -C(O)NH-.

37. The antibody drug conjugate of any one of claims 32-36, wherein, Q' is selected from: represents a connection site to L5, represents a connection site to L4.

38. The antibody drug conjugate of claim 37, wherein, L5-Q' is selected from: L5-Q' is selected from: L5-Q' is selected from:

39. The antibody drug conjugate of claim 38, wherein, L5-Q' is selected from Preferably, Z1and Z2are each independently selected from -C(O)NH-.

40. The antibody drug conjugate of any one of claims 32-39, wherein, Ab is selected from a murine antibody, a chimeric antibody, a humanized antibody, a fully human antibody, or an antigen-binding fragment thereof; preferably, the Ab is selected from an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-ROR1 antibody, an anti-CLDN6 antibody, an anti-CLDN9 antibody, an anti-CLDN18.2 antibody, an anti-NaPi-2b antibody, an anti-TNF-a antibody, an anti-ENPP3 antibody, an anti-DLL3 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD28 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD37 antibody, an anti-CD38 antibody, an anti-CD44 antibody, an anti-CD45 antibody, an anti-CD47 antibody, an anti-CD48 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD98 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-EphA2 antibody, an anti-MUC1 antibody, an anti-Lewis Y antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, an anti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-C antibody, an anti-SLC44A4 antibody, an anti-CD79 antibody, an anti-TROP-2 antibody, an anti-CD79B antibody, an anti-Mesothelin antibody, an anti-Nectin-4 antibody, an anti-TPBG antibody, or an antigen-binding fragment thereof.

41. The antibody drug conjugate of claim 40, wherein, The Ab is an anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof.

42. The antibody drug conjugate of claim 40, wherein, Ab is selected from a monoclonal antibody or an antigen-binding fragment thereof, preferably from Trastuzumab, Cetuximab, Pertuzumab, Nimotuzumab, Enoblituzumab, Emibetuzumab, Inotuzumab, Pinatuzumab, Brentuximab, Gemtuzumab, Bivatuzumab, Lorvotuzumab, or an antigen-binding fragment thereof.

43. The antibody drug conjugate of any one of claims 32-42, wherein, Formula (I") is one of the following: ; preferably, the Ab is an anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof; more preferably, Trastuzumab or an antigen-binding fragment thereof.

44. An eribulin derivative which is a compound as shown below, or an isotopically- labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof, 45. An antibody drug conjugate, which is a compound as shown below or an isotopically-labeled compound thereof, or an optical isomer, a geometric isomer, a tautomer, or an isomer mixture thereof, or a pharmaceutically acceptable salt thereof, wherein, Ab is an antibody or an antigen-binding fragment; n is an integer or a decimal number between 0 and 20; preferably, n is an integer or a decimal number between 0 and 8.

46. The antibody drug conjugate of claim 45, wherein, Ab is selected from a murine antibody, a chimeric antibody, a humanized antibody, a fully human antibody, or an antigen-binding fragment thereof; preferably, the Ab is selected from an anti-HER2 (ErbB2) antibody, an anti-EGFR antibody, an anti-B7-H3 antibody, an anti-c-Met antibody, an anti-HER3 (ErbB3) antibody, an anti-HER4 (ErbB4) antibody, an anti-ROR1 antibody, an anti-CLDN6 antibody, an anti-CLDN9 antibody, an anti-CLDN18.2 antibody, an anti-NaPi-2b antibody, an anti-TNF-a antibody, an anti-ENPP3 antibody, an anti-DLL3 antibody, an anti-CD20 antibody, an anti-CD22 antibody, an anti-CD28 antibody, an anti-CD30 antibody, an anti-CD33 antibody, an anti-CD37 antibody, an anti-CD38 antibody, an anti-CD44 antibody, an anti-CD45 antibody, an anti-CD47 antibody, an anti-CD48 antibody, an anti-CD56 antibody, an anti-CD70 antibody, an anti-CD73 antibody, an anti-CD98 antibody, an anti-CD105 antibody, an anti-CEA antibody, an anti-EphA2 antibody, an anti-MUC1 antibody, an anti-Lewis Y antibody, an anti-VEGFR antibody, an anti-GPNMB antibody, an anti-Integrin antibody, an anti-PSMA antibody, an anti-Tenascin-C antibody, an anti-SLC44A4 antibody, an anti-CD79 antibody, an anti-TROP-2 antibody, an anti-CD79B antibody, an anti-Mesothelin antibody, an anti-Nectin-4 antibody, an anti-TPBG antibody, or an antigen-binding fragment thereof.

47. The antibody drug conjugate of claim 46, wherein, The Ab is an anti-HER2 (ErbB2) antibody or an antigen-binding fragment thereof.

48. The antibody drug conjugate of claim 46, wherein, Ab is selected from a monoclonal antibody or an antigen-binding fragment thereof, preferably from Trastuzumab, Cetuximab, Pertuzumab, Nimotuzumab, Enoblituzumab, Emibetuzumab, Inotuzumab, Veltuzumab, Pinatuzumab, Brentuximab, Gemtuzumab, Bivatuzumab, Lorvotuzumab, or an antigen-binding fragment thereof; preferably, the Ab is Trastuzumab or an antigen-binding fragment thereof.

49. A pharmaceutical composition comprising the eribulin derivative of any one of claims 1-31, 44, or the antibody drug conjugate of any one of claims 32-43, 45-48, and one or more pharmaceutically acceptable carriers or excipients.

50. Use of the eribulin derivative of any one of claims 1-31, 44 in the manufacture of an antibody drug conjugate.

51. Use of the eribulin derivative of any one of claims 1-31, 44, or the antibody drug conjugate of any one of claims 32-43, 45-48, or the pharmaceutical composition of claim 49 in the manufacture of a medicament for treating a tumor or cancer.

52. A method of treating a tumor or cancer, comprising administering to a subject in need thereof an effective amount of the eribulin derivative of any one of claims 1-31, 44, or the antibody drug conjugate of any one of claims 32-43, 45-48, or the pharmaceutical composition of claim 49.

53. The use according to claim 51, or the method according to claim 52, characterized in that, the tumor or cancer is selected from a solid tumor or a hematological tumor; preferably from breast cancer, gastric cancer, colon cancer, pancreatic cancer, renal cancer, ovarian cancer, lung cancer, soft tissue sarcoma, liposarcoma, lung cancer, non-small cell lung cancer, melanoma, head and neck cancer, cervical cancer, prostate cancer; more preferably from HER-2 positive breast cancer, triple negative breast cancer, HER-2 positive triple negative breast cancer, renal clear cell adenocarcinoma, small cell lung cancer, non-small cell lung cancer, large cell lung cancer.

54. The use or method of claim 53, wherein, the tumor or cancer is selected from a cancer or tumor with high expression of HER2 (ErbB2) antibody.

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