C9-substituted camptothecin derivative and conjugate thereof

Abstract

Provided are a C9-substituted camptothecin derivative and a conjugate thereof. Specifically disclosed is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The camptothecin derivative and the conjugate thereof have a good cell killing effect.

Description

C9-substituted camptothecin derivatives and their conjugates

[0001] This application claims priority to Chinese Patent Application No. 2024119169566, filed on December 24, 2024, and Chinese Patent Application No. 2025114825099, filed on October 16, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention relates to a C9-substituted camptothecin derivative and its conjugates. Background Technology

[0003] Malignant tumors are among the diseases that seriously affect human health. According to the International Agency for Research on Cancer (IARC) of the World Health Organization, there were 20 million new cancer cases and 9.7 million deaths worldwide in 2022.

[0004] Antibody-drug conjugates (ADCs) are a novel type of cancer treatment. They use highly cytotoxic small molecules as payloads, which are covalently linked to antibodies via linkers. These antibodies can selectively bind to antigens highly expressed in tumor tissue, thereby delivering the payload with high specificity to the tumor tissue and achieving selective killing of tumor cells.

[0005] Camptothecin derivatives are currently the most widely used toxic loads in the ADC field. These compounds inhibit topoisomerase I, thereby blocking gene replication and expression processes in cells and inducing cell death. Their main chemical structure consists of five six-membered rings (A, B, C, D, and E). The E-ring is a six-membered lactone, which is the biologically active center of camptothecin compounds. This ring is closed under neutral and acidic conditions. Under alkaline conditions, the lactone hydrolyzes and opens, losing its biological activity. Under acidic conditions, it closes again, regaining its activity. Therefore, camptothecin compounds exist in an aqueous solution in a pH-dependent equilibrium; the stronger the alkalinity, the greater the proportion of inactive compounds existing in their open-ring form. Daiichi Sankyo developed the camptothecin derivative Exatecan (whose mesylate salt, as a small molecule anticancer drug, failed to meet the therapeutic endpoint in a Phase III clinical trial, leading to the termination of further development). By adding a six-membered ring (F-ring) between C9 of the A-ring and C7 of the B-ring in the main structure of camptothecin, Daiichi Sankyo discovered that the F-ring has a stabilizing effect on the E-ring, making it less susceptible to hydrolysis. The activity of Exatecan is more than 10 times higher than that of SN-38 (the in vivo active compound of the approved drug irinotecan). Daiichi Sankyo used the Exatecan derivative Dxd as a payload and conjugated it with trastuzumab, which targets HER2, via the tetrapeptide chain GGFG to develop the ADC drug ENHERTU (also known as DS-8201 or T-Dxd), achieving effective treatment for HER2-positive breast cancer, gastric cancer, esophageal junction cancer, HER2-low-expressing breast cancer, and non-small cell lung cancer.

[0006] Despite its proven success as one of the most successful ADCs in history, DS-8201, as a first-generation camptothecin-based ADC, still has several shortcomings that urgently need to be addressed. First, natural or acquired resistance to DS-8201 has been observed in clinical applications. Second, DS-8201 exhibits poor plasma stability, making it prone to releasing toxins in non-tumor tissues, leading to hematologic toxicity. Finally, interstitial lung disease (ILD) is a key adverse reaction identified in DS-8201 clinical studies. In the pivotal Phase II Destination-Breast01 study (NCT03248492), 13.6% of participants developed ILD; while in the pivotal Phase II DISTY-GASTEC01 study (NCT03329690), 12 out of 119 patients exhibited DS-8201-related ILD or pneumonia, with one patient dying from pneumonia.

[0007] Currently, most ADC payloads developed based on camptothecin are derivatives of ixotecan, retaining the F ring that stabilizes the E ring. However, this is not an advantage for ADC payloads because if the ADC drug releases camptothecin toxin due to linker breakage in the bloodstream, an overly stable payload can cause strong side effects. Ideally, under physiological conditions (pH 7.4), the E ring of camptothecin can undergo hydrolysis to open, forming an inactive metabolite, thereby reducing systemic toxicity caused by the toxin. Furthermore, the E ring can remain stable in the lysosomes of tumor cells (pH 4.5–5.5), thus exhibiting cytotoxic activity. In summary, the ease with which the E ring is hydrolyzed in an alkaline environment is a promising advantage for ADC payloads, potentially reducing systemic toxicity in clinical applications. Considering the safety issues exhibited by DS-8201 in clinical use, improving the linker-payload of camptothecin is essential. Summary of the Invention

[0008] The technical problem to be solved by this invention is to overcome the single structure of existing camptothecin derivatives and their conjugates, and to provide a C9-substituted camptothecin derivative and its conjugate. The camptothecin derivative and its conjugate provided by this invention have a novel structure, are directly modified at the C9 position of the A ring, and lack the F ring, thus exhibiting better cell-killing activity.

[0009] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof;

[0010] Among them, R 1 and R 2 Same or different, and independently hydrogen, hydroxyl, mercapto, halogen, cyano, -NR c R d C 1-6 Alkyl group, -(CH2CH2O) m R c C 1-6 Alkoxy, C 1-6 Alkylthio, -C(=O)NR c R d -NR c C(=O)R d -SOR c or -SO2R c The C 1-6 Alkyl group, -(CH2CH2O) m R c C 1-6 Alkoxy, C 1-6 Alkylthio, -C(=O)NR c R d-NR c C(=O)R d -SOR c and -SO2R c Optionally substituted with halogen, hydroxyl or amino groups;

[0011] Or, R 1 and R 2 Together Formation - XL a -Y- ring; X and Y are each independently selected from -NH- or -S-, L a C 1-3 Alkylene or C 1- 3-Halogenide;

[0012] Alk is C 2-6 imidene group, C 2-6 Ethyne group, C 5-8 Cycloalkylene, C 6-10 arylene, 5-10-membered heterocyclic alkenyl or 5-10-membered heteroarylene, wherein C 2-6 imidene group, C 2-6 Ethyne group, C 5-8 Cycloalkylene, C 6-10 arylene, 5-10-membered heterocyclic alkenyl and 5-10-membered heteroarylene are optionally surrounded by 1-8 R a replace;

[0013] R a Independently, it can be halogen, cyano, nitro, hydroxyl, mercapto, oxo (=O), or -NR. c R d Carboxyl group, -C(=O)NR c R d -N(R) c )C(=O)R d -SO2NR c R d -N(R) c SO2R d C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, -(CH2CH2O) m R c C 3-8 Cycloalkyl, 3-8 membered heterocycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, -SOR c -SO2R c -P(=O)R c R d -NR cC(=O)NR c R d or -C(=O)R c ;

[0014] Q 1 The following are non-existent: -NH-, -O-, -S-, -C(=O)-, -C(=O)NH-, -NHC(=O)-, -C(=O)O-, -OC(=O)-, -S(=O)-, -SO2-, -SO2NH-, -NHSO2-, -S(=O)-NH-, -NH-S(=O)-, -P(=O)R c -, -NHS(=O)NH-, -NHC(=O)NH-, -NHC(=S)NH- or -NHC(=NH)NH-;

[0015] L and L b Independently non-existent, C 1-6 Alkylene, C 2-6 imidene group, C 2-6 Alynyl group, -(CH2CH2O) m -、-OC 1-6 Alkylene, -N(C) 1-6 alkyl)-, -SC 1-6 Alkylene, C 3-8 Cycloalkylene or 3-8 membered heterocycloalkylene; the C 1-6 Alkylene, C 2-6 imidene group, C 2-6 Alynyl group, -(CH2CH2O) m -、-OC 1-6 Alkylene, -N(C) 1-6 alkyl)-, -SC 1-6 Alkylene, C 3-8 Cycloalkylene and 3-8-membered heterocycloalkylene are optionally halogenated or C 3-8 Cycloalkyl substitution; and L b When it does not exist, Q 1 It does not exist;

[0016] m is an independent integer from 1 to 10;

[0017] Q 2 It is hydrogen, hydroxyl, mercapto, or amino; the amino group is optionally C 1-6 Alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl or C 2-6 Alkyne substitution;

[0018] R c and R d Independently hydrogen or C 1-6 alkyl;

[0019] The heteroatoms in the 5-10 member heterocyclic alkenyl group, 5-10 member heteroaryl group, and 3-8 member heterocyclic alkyl group are one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three.

[0020] In some embodiments, in the compound represented by formula (I) or its pharmaceutically acceptable salt, some groups are defined as follows, and the remaining groups are defined as in any other embodiment (hereinafter referred to as "in some embodiments"): R 1 and R 2 Same or different, and independently hydrogen, hydroxyl, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl groups, -NR c R d , thiol, C 1-6 Alkoxy, C 1-6 Alkylthio, -S(=O)R c or -SO2R c Among them, R c C 1-6 Alkyl; R d It is hydrogen or C 1-6 alkyl.

[0021] In some implementation schemes, R 1 and R 2 They may be the same or different, and are independently hydrogen, hydroxyl, fluorine, cyano, methoxy, methyl or trifluoromethyl.

[0022] In some implementation schemes, R 1 C 1-6 alkyl.

[0023] In some implementation schemes, R 1 It is -CH3.

[0024] In some implementation schemes, R 2 It is a halogen.

[0025] In some implementation schemes, R 2 It is -F.

[0026] In some implementations, Alk is C 2-6 imidene group, C 5-8 Cycloalkylene, C 2-6 Alynyl, phenylene, 5-6 membered heterocyclic alkenyl or 5-6 membered heteroaryl, wherein C 2-6 imidene group, C 5-8 Cycloalkylene, C 2-6 The alkynyl, phenylene, 5-6 membered heterocyclic alkenyl or 5-6 membered heteroaryl group is optionally surrounded by 1-8 R groups. aSubstitution; the heteroatoms in the 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl are one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0027] In some implementations, Alk is -CH=CH-, Cyclohexene, phenylene Pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thiopheneyl, pyrroloyl, pyrazolyl, imidazoyl, oxazolyl, isoxazolyl, thiazolyl, or isothiazolyl; wherein the cyclohexeneyl, phenylene, Pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thiopheneyl, pyrroloyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl are optionally surrounded by 1-4 R groups. a replace.

[0028] In some implementations, Alk is -CH=CH-, Cyclohexene, phenylene Pyridyl or imidazolyl, wherein the phenylene, pyridyl, and imidazolyl groups are optionally surrounded by 1-2 R groups. a replace.

[0029] In some implementation schemes, R a Independently, it can be a halogen, cyano group, oxo group (=O), or C group. 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkyl groups.

[0030] In some implementation schemes, R a It can be fluorine, cyano, oxo (=O), methyl, methoxy, or trifluoromethyl independently.

[0031] In some implementations, Alk is End a is connected to L.

[0032] In some implementation schemes, Q 1 For non-existent, -NH-, -O-, b -NHC(=O)- or b -C(=O)NH-, the b end is connected to L.

[0033] In some implementation schemes, Q 1 For non-existent or b -NHC(=O)-, the b end is connected to L.

[0034] In some implementations, L and L bIndependently non-existent, C 1-6 Alkylene, C 1-6 Halogenated alkylene or C 3-8 Cycloalkyl-substituted C 1- 6-alkylene oxide.

[0035] In some implementations, L and L b Independently for non-existence or C 1-6 Alkylene.

[0036] In some implementations, L is not present.

[0037] In some implementations, L b It is non-existent, -CH2- or -CH2-CH2-.

[0038] In some implementation schemes, Q 2 It is hydrogen, hydroxyl, amino, or C 1-6 Alkyl-substituted amino groups.

[0039] In some implementation schemes, Q 2 It can be hydrogen, hydroxyl, amino, or methylamino.

[0040] In some implementation schemes, R 1 C 1-6 alkyl;

[0041] R 2 It is a halogen;

[0042] Alk is C 2-6 Ethyne group, C 5-8 Cycloalkylene, C 6-10 arylene, 5-10-membered heterocyclic alkenyl or 5-10-membered heteroarylene, wherein C 2-6 Ethyne group, C 5-8 Cycloalkylene, C 6-10 arylene, 5-10-membered heterocyclic alkenyl and 5-10-membered heteroarylene are optionally surrounded by 1-2 R a replace;

[0043] R a Independently halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkyl groups;

[0044] Q 1 For non-existent, -NH-, -O-, b -NHC(=O)- or b -C(=O)NH-, the b end is connected to L;

[0045] L means it does not exist;

[0046] L b For non-existent or C 1-6 Alkylene;

[0047] And L b When it does not exist, Q 1 It does not exist;

[0048] Q 2 It is hydrogen, hydroxyl, amino, or C 1-6 Alkyl-substituted amino groups.

[0049] In some embodiments, the compound represented by formula (I) is any one of the compounds in Table A below:

[0050] Table A

[0051] The present invention also provides a pharmaceutical composition comprising a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

[0052] The present invention also provides the use of a compound of formula (I) as described above, or a pharmaceutically acceptable salt thereof, or a composition as described above, in the preparation of a medicament for the prevention or treatment of a disease.

[0053] In some implementations, the disease is a disease related to abnormal cellular activity.

[0054] In some implementations, the diseases associated with abnormal cell activity are esophageal cancer, brain tumors, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial carcinoma, solid tumors, non-Hodgkin's lymphoma, leukemia, central nervous system tumors, prostate cancer, thyroid cancer, or large B-cell lymphoma.

[0055] The present invention also provides a compound of formula (II) or a pharmaceutically acceptable salt thereof.

[0056] L 1 -D (II)

[0057] Among them, L 1 For connecting precursors;

[0058] D is a group formed by the loss of a hydrogen atom in the compound shown in formula (I) above.

[0059] In this invention, the linker precursor refers to a degradable or non-degradable linker that can react with an antibody to form a linker for connecting D and the antibody.

[0060] In some embodiments, in the compound of formula (II) or its pharmaceutically acceptable salt, some groups are defined as follows, and the remaining groups are defined as in any other embodiment (hereinafter referred to as "in some embodiments"):

[0061] L 1 For MM 1 -L c -C(=O)-AA-SE-;

[0062] Among them, MM 1 It is hydrogen or an electrophilic group;

[0063] L c It does not exist; -(CH2) q -、 c -(CH2) q1 C(=O)N(R e (CH2) q2 -、 c -(CH2) q1 (OCH2CH2) q2 -or c -(CH2) q1 (OCH2CH2) q2 N(R e )C(=O)(CH2) q1 -, C-end and MM 1 connect;

[0064] R e It is hydrogen or C 1-6 alkyl;

[0065] q and q1 are independent integers from 1 to 10; q2 is an integer from 1 to 100;

[0066] AA is a polypeptide residue consisting of 2-10 amino acids, which may be the same or different, and optionally converted by one or more polyols or C. 1-6 Alkyl substitution; the C-terminus of the polypeptide residues is linked to SE, and the N-terminus is linked to -C(=O)-;

[0067] SE indicates that it does not exist. Connect end 1 to AA, and end 2 to D;

[0068] R 3 For hydrogen or -L d -Q 3 -L e -MM 2 ;

[0069] L d For does not exist, C1-10 Alkylene or C 3-10 Cycloalkylene;

[0070] Q 3 For -NR f -, -O-, -S-, -C(=O)-, -N(R f (C=O)-、-C(=O)N(R f )-, -OC(=O)-, -C(=O)O-, -S(=O)-, -SO2-, -SO2NR f -、-NR f SO2-, -S(=NH)(=O)-, -P(=O)R f -, -NHC(=O)NH-, -NHC(=S)NH- or -NHC(=NH)NH-;

[0071] L e It does not exist; -(CH2) p -、 d -(CH2) p1 C(=O)N(R g (CH2) p2 -、 d -(CH2) p1 (OCH2CH2) p2 N(R g )C(=O)(CH2) p1 -、 d -(CH2) p1 C(=O)N(R g )CH2CH2(OCH2CH2) p2 -or d -(CH2) p1 (OCH2CH2) p2 -,d end and MM 2 Covalent connection;

[0072] R f and R g Independently hydrogen or C 1-6 alkyl;

[0073] p and p1 are independent integers from 1 to 10; p2 is an integer from 1 to 100;

[0074] MM 2 It is an electrophilic group;

[0075] R 3 With MM 1 There is exactly one hydrogen atom.

[0076] In some implementations, L c forc -(CH2) q3 C(=O)N(R e (CH2CH2O) q4 (CH2) q5 -or c -(CH2CH2O) q6 (CH2) q7 -, C-end and MM 1 Connection; R e It is hydrogen or C 1-6 Alkyl group; q3, q4, q5, q6 and q7 are independently integers from 1 to 10.

[0077] In some implementations, q6 and q7 are independently 1, 2, 3, 4, 5, or 6.

[0078] In some implementations, D is

[0079] Q 2a For non-existent, -O-, -NH-, or -N(C) 1-6 alkyl)-;

[0080] R 1 R 2 Alk, L, Q 1 and L b The definition is as described above.

[0081] In some implementation schemes, Q 2a It is non-existent, -O-, -NH-, or -N(CH3)-.

[0082] In some implementations, MM 1 Hydrogen, halogen, Michael acceptor group, epoxy group, R h Independently hydrogen or C 1-6 alkyl.

[0083] In some implementations, MM 1 For hydrogen, halogen, R h Independently hydrogen or C 1-6 alkyl.

[0084] In some implementations, MM 1 For hydrogen, -Br or

[0085] In some implementations, L c -(CH2) q -、 c-CH2C(=O)NHCH2CH2-、 c -CH2CH2OCH2CH2OCH2CH2NHC(=O)CH2CH2-, c-terminus with MM 1 Connect; q is 1-6.

[0086] In some implementations, L c for c -(CH2) q3 C(=O)NH(CH2CH2O) q4 (CH2) q5 -, C-end and MM 1 Connect; q3, q4, and q5 are independently 1, 2, 3, 4, 5, or 6.

[0087] In some implementations, L c It is -CH2-, -CH2CH2-, or -CH2-CH2-CH2-CH2-CH2-.

[0088] In some implementations, L c for c -CH2C(=O)NHCH2CH2OCH2CH2OCH2- or c -(CH2CH2O)2-CH2-, c-terminus and MM 1 connect.

[0089] In some embodiments, AA is a polypeptide residue consisting of 2, 3, or 4 amino acids, which may be the same or different, and optionally by one or more amino acids. Or C 1-6 Alkyl substitution; the C-terminus of the polypeptide residue is linked to SE, and the N-terminus is linked to -C(=O)-.

[0090] In some embodiments, the amino acid is D-type or L-type.

[0091] In some embodiments, the amino acid is glycine, alanine, valine, leucine, isoleucine, methionine, proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, citrulline, or one or more of the following. Or C 1-6 Alkyl-substituted glutamine, with one or more Or C 1- 6-alkyl-substituted glutamic acid, or glutamic acid with one or more C atoms 1-6 Alkyl-substituted lysine.

[0092] In some embodiments, the amino acid is Ri Independently H or C 1-6 alkyl.

[0093] In some embodiments, AA is -Val-Ala-、-Ala-Val-、-Gly-Gly-、-Val-Cit-、-Cit-Val-、-Glu-Ala-、-Glu-Lys-、-Glu-Gly-、-Glu-Cit-、-Leu-Ala-、-Ala-Leu-、-L eu-Cit-、-Cit-Leu-、-Lys-Lys-、-Ala-Lys-、-Lys-Ala-、-Val-Lys-、-Lys-Val-、-Tyr-Arg-、-Arg-Tyr-、-Arg-Arg-、-Ala-Ala-、-Phe-Lys-、-Lys-Ph e-、-Thr-Thr-、-Thr-Met-、-Met-Thr-、-Met-Tyr-、-Tyr-Met-、-Phe-Gln-、-Gln-Phe-、-Gly-Ser-、-Leu-Gln-、-Gln-Leu-、-Ser-Ala-、-Ser-Gly-、-V al-Thr-、-Thr-Val-、-Val-Gln-、-Ser-Val-、-Val-Ser-、-Ala-Met-、-Met-Ala-、-Val-Arg-、-Arg-Val-、-Phe-Ala-、-Ala-Phe-、-Gln-Val-、-Phe-Ar g-、-Arg-Phe-、-Glu-Gly-Cit-、-Ala-Ala-Ala-、-Gly-Gly-Gly-、-Ala-Val-Ala-、-Gly-Val-Gly-、-Ala-Val-Gly-、-Gly-Phe-Lys-、-Lys-Phe-Gly-、 -Leu-Ala-Leu-、-Val-Ala-Leu-、-Leu-Val-Ala-、-Val-Ala-Val-、-Gly-Lys-Val-、-Gly-Lys-Gly-、-Val-Lys-Gly-、-Glu-Gly-Cit-、-Glu-Val-Cit- 、-Gln-Val-Cit-、-Glu-Val-Ala-、-Gln-Val-Ala-、-Glu-Lys-Gly-、-Glu-Phe-Gly-、-Ala-Val-Ala-Gly-、-Gly-Phe-Gly-Gly-、-Gly-Gly-Phe-Gly-、 -Ala-Val-Gly-Gly-、-Gly-Gly-Val-Ala、-Ala-Ala-Ala-Ala-、-Ala-Leu-Ala-Leu-、-Leu-Ala-Leu-Ala-、-Gly-Phe-Leu-Gly-、-Gly-Leu-Phe-Gly-、-Glu-Glu-Lys-Gly-, -Glu-Glu-Val-Cit-, -Gln-Glu-Val-Cit-, -Glu-Glu-Val-Ala-, -Gln-Glu -Val-Ala-, -Gly-Glu-Lys-Gly-, -Gly-Glu-Phe-Gly-, -Gln-Gly-Phe-Gly-, -Glu-Glu-Gly-Cit, The C terminal of AA is connected to SE, and the N terminal is connected to -C (=O)-.

[0094] In some implementations, AA is -Ala-Ala-Asn- or Gly-Ala-Asn-, with the C terminal of AA connected to SE and the N terminal connected to -C(=O)-.

[0095] In some implementation schemes, AA is The C terminal of AA is connected to SE, and the N terminal is connected to -C (=O)-.

[0096] In some implementation schemes, AA is The C terminal of AA is connected to SE, and the N terminal is connected to -C (=O)-.

[0097] In some implementations, L d For non-existent or C 1-6 Alkylene.

[0098] In some implementations, L d It does not exist.

[0099] In some implementation schemes, Q 3 -NH(C=O)- e -C(=O)NH- e or -C(=O)O- e e-end and L e connect.

[0100] In some implementation schemes, Q 3 -C(=O)NH- e e-end and L e connect.

[0101] In some implementations, L e -(CH2) p -、 d -(CH2) p1C(=O)NH(CH2) p2 -、 d -(CH2) p1 (OCH2CH2) p2 NHC(=O)(CH2) p1 -、 d -(CH2) p1 C(=O)NHCH2CH2(OCH2CH2) p2 -or d -(CH2) p1 (OCH2CH2) p2 -,d end and MM 2 Covalent connection.

[0102] In some implementations, p and p1 are independently integers from 1 to 6, such as 1 or 5.

[0103] In some implementations, p2 is an integer from 1 to 10, such as 2.

[0104] In some implementations, L e -(CH2) p -or d -(CH2) p1 C(=O)NH(CH2) p2 -,d end and MM 2 Covalent connection; p and p1 are independent integers from 1 to 6, and p2 is an independent integer from 1 to 6.

[0105] In some implementations, L e -(CH2)6- or d -CH2-C(=O)NH(CH2)2-, d-terminus with MM 2 Covalent connection.

[0106] In some implementations, MM 2 Hydrogen, halogen, Michael acceptor group, epoxy group, R j Independently hydrogen or C 1-6 alkyl.

[0107] In some implementations, MM 2 For -Br or

[0108] In some implementation schemes, R 3 for

[0109] In some implementations, SE is non-existent. Connect end 1 to AA and end 2 to D.

[0110] In some implementations, D is

[0111] Q 2a For non-existent, -O-, -NH-, or -N(C) 1-6 alkyl)-;

[0112] L 1 For MM 1 -L c -C(=O)-AA-SE-;

[0113] MM 1 Hydrogen, halogen or

[0114] L c -(CH2) q -, q is 1-6;

[0115] AA is a polypeptide residue consisting of 2, 3, or 4 amino acids, which may be the same or different, and optionally divided by one or more amino acids. Or C 1-6 Alkyl substitution; the C-terminus of the polypeptide residues is linked to SE, and the N-terminus is linked to -C(=O)-;

[0116] SE indicates that it does not exist. Connect end 1 to AA, and end 2 to D;

[0117] R 3 For hydrogen or -L d -Q 3 -L e -MM 2 ;

[0118] L d It does not exist;

[0119] Q 3 -NH(C=O)- e -C(=O)NH- e or -C(=O)O- e e-end and L e connect;

[0120] L e -(CH2) p -、 d -(CH2) p1 C(=O)NH(CH2) p2 -,d end and MM 2 Covalent connection;

[0121] p and p1 are independent integers from 1 to 6; p2 is an integer from 1 to 10;

[0122] MM 2 Hydrogen, halogen or

[0123] R 3 With MM 1 There is one and only one hydrogen atom;

[0124] R 1 R 2 Alk, L, Q 1 and L b The definition is as described above.

[0125] In some implementations, D is R 1 C 1-6 alkyl;

[0126] R 2 It is a halogen;

[0127] L 1 For MM 1 -L c -C(=O)-AA-SE-;

[0128] MM 1 It is a halogen;

[0129] L c -(CH2) q -or c -(CH2) q3 C(=O)NH(CH2CH2O) q4 (CH2) q5 -, C-end and MM 1 Connect; q is 1-6; q3, q4, and q5 are independently 1, 2, 3, 4, 5, or 6;

[0130] AA is a polypeptide residue consisting of 2, 3, or 4 amino acids, which may be the same or different, and optionally divided by one or more amino acids. Or C 1-6 Alkyl substitution; the C-terminus of the polypeptide residues is linked to SE, and the N-terminus is linked to -C(=O)-;

[0131] SE is Connect end 1 to AA and end 2 to D.

[0132] In some embodiments, the compound represented by formula (II) is any one of the compounds in Table B below:

[0133] Table B

[0134] This invention provides a conjugate of formula (III) or a pharmaceutically acceptable salt thereof;

[0135] Ab-[L 2 -D]ρ (III)

[0136] Wherein, Ab is an antibody or its antigen-binding fragment;

[0137] L 2 For connectors;

[0138] ρ is the drug loading, and can be any value between 1 and 20;

[0139] The definition of D is as described above.

[0140] In some implementations, L 2 for f -MM 3 -L c -C(=O)-AA-SE-or The f terminal is connected to Ab.

[0141] SE indicates that it does not exist. Connect end 1 to AA, and end 2 to D;

[0142] MM 1 It is hydrogen;

[0143] R3a -L d -Q 3 -L e -MM 4 - f The f end is connected to Ab;

[0144] MM 3 and MM 4 Independently for non-existence or

[0145] L c AA, L d Q 3 and L e The definition is as described above.

[0146] In some embodiments, the antibody or its antigen-binding fragment includes Fab, Fab', F(ab')2, Fd, Fv, dAb, complementarity-determining region fragment, single-chain antibody (e.g., scFv), non-human antibody, humanized antibody, chimeric antibody, fully human antibody, probody, monoclonal antibody, bispecific antibody, or heterologous antibody.

[0147] In some implementations, Ab is an antibody or its antigen-binding fragment that has endocytosis, non-endocytosis, or weak endocytosis.

[0148] In some implementations, Ab is an antibody or its antigen-binding fragment that has the activity of binding free antigens in tumor tissue and / or tumor surface antigens.

[0149] In some implementations, Ab is an antibody or its antigen-binding fragment that does not have the activity of binding to free antigens in tumor tissue and / or tumor surface antigens.

[0150] In some implementations, Ab is an antibody or antigen-binding fragment thereof targeting HER2 (ErbB2), HER3 (ErbB3), HER4 (ErbB4), EGFR, B7H3, TROP2, CD19, CD79b, CD33, FOLR1, or Nectin4.

[0151] In some implementations, Ab is an antibody or antigen-binding fragment targeting HER2.

[0152] In some embodiments, the Ab is an anti-HER2 antibody or its antigen-binding fragment, such as anbenitamab, coprelotamab, disitamab, gancotamab, margetuximab, timigutuzumab, zanidatamab, trastuzumab, pertuzumab, inetetamab or its antigen-binding fragment; more preferably, the Ab is trastuzumab or pertuzumab.

[0153] In some implementations, Ab is trastuzumab.

[0154] In some implementations, Ab is an anti-EGFR antibody or its antigen-binding fragment, such as demupitamab, depatuxizumab, futuximab, imgatuzumab, lapatuximab, losatuxizumab, matuzumab, modotuximab, necitumumab, nimotuzumab, panitumumab, pimurutamab, serclutamab, tomuzotuximab, zalutumumab, Cetuximab, intetumumab, lapatuximab, or its antigen-binding fragment.

[0155] In some implementations, Ab is an anti-B7H3 antibody or its antigen-binding fragment, such as enoblituzumab, mirzotamab, omburtamab, ifinatamab, vobramitamab antibody or its antigen-binding fragment.

[0156] In some implementations, Ab is an anti-TROP2 antibody or its antigen-binding fragment, such as datopotamab, sacituzumab or its antigen-binding fragment.

[0157] In some implementations, ρ is any value between 1 and 10, preferably any value between 2 and 8.

[0158] In some implementations, ρ is an integer between 1 and 10, such as 1, 2, 3, 4, 5, 6, 7, or 8.

[0159] In some implementations, ρ is a decimal between 1 and 10, preferably a decimal between 2 and 8.

[0160] In some implementations, ρ is any value between 2 and 9, such as 4.0, 7.7, 7.8, 7.9, 7.95, 8.0, 8.02, 8.05, 8.06, 8.15, or 8.2.

[0161] In some embodiments, in the conjugate represented by formula (III), D is R 1 C 1- 6-alkyl;

[0162] R 2 It is a halogen;

[0163] L 2 for f -MM 3 -L c -C(=O)-AA-SE-, f end is connected to Ab;

[0164] MM 3 It does not exist;

[0165] L c -(CH2) q -or c -(CH2) q3 C(=O)NH(CH2CH2O) q4 (CH2) q5 -, C-end and MM 3 Connect; q is 1-6; q3, q4, and q5 are independently 1, 2, 3, 4, 5, or 6;

[0166] AA is a polypeptide residue consisting of 2, 3, or 4 amino acids, which may be the same or different, and optionally divided by one or more amino acids. Or C 1-6 Alkyl substitution; the C-terminus of the polypeptide residues is linked to SE, and the N-terminus is linked to -C(=O)-;

[0167] SE is Connect end 1 to AA, and end 2 to D;

[0168] Ab is an antibody or its antigen-binding fragment, preferably trastuzumab;

[0169] ρ can be any value between 1 and 10.

[0170] In some embodiments, the conjugate represented by formula (III) is any of the conjugates in Table C below:

[0171] Table C

[0172] The definitions of Ab and ρ are as described above.

[0173] In some embodiments, the conjugate represented by formula (III) is any of the following conjugates:

[0174] The present invention also provides a pharmaceutical composition comprising the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

[0175] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described above, in the preparation of a medicament for the prevention or treatment of a disease. The disease is preferably a disease related to abnormal cellular activity, such as esophageal cancer, brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial carcinoma, solid tumors, non-Hodgkin's lymphoma, leukemia, central nervous system tumors, prostate cancer, thyroid cancer, or large B-cell lymphoma.

[0176] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of HER2-related, mediated diseases, such as breast cancer.

[0177] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of HER3-related, mediated diseases, such as breast cancer or lung cancer.

[0178] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of HER3-related, mediated diseases, such as breast cancer or lung cancer.

[0179] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of TROP2-related, mediated diseases, such as breast cancer, lung cancer, ovarian cancer or endometrial cancer.

[0180] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of CD19-related, mediated diseases, such as large B-cell lymphoma.

[0181] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of CD79b-related, mediated diseases, such as non-Hodgkin lymphoma.

[0182] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of FOLR1-related, mediated diseases, such as ovarian cancer or peritoneal cancer.

[0183] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of Nectin4-related, mediated diseases, such as esophageal cancer or breast cancer.

[0184] The present invention also provides the use of the conjugate of formula (III) as described above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention or treatment of CD33-related, mediated diseases, such as leukemia.

[0185] Abbreviations and Definitions

[0186] Unless otherwise stated, the following terms as used in this application have the following definitions.

[0187] Unless otherwise specified, the term "conjugate" refers to the covalent bonding of two identical or different chemical structures to form a new chemical structure. This includes, but is not limited to, antibody-drug conjugates.

[0188] Unless otherwise specified, the term "antibody-drug conjugate" refers to a drug formed by covalently linking an antibody or antigen-binding fragment to a biologically active cytotoxic drug.

[0189] The term "geometric isomer" refers to isomers that exist in a molecule but have different spatial arrangements of atoms or groups due to the presence of double bonds or rings. For example, cis-2-butene and trans-2-butene are geometric isomers of each other.

[0190] The term "optical isomer" refers to isomers that, due to their chiral centers or chiral axes, can deflect polarized light in different directions. This includes, but is not limited to, enantiomers and diastereomers. For example, L-alanine and D-alanine are optical isomers of each other.

[0191] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound (e.g., a drug, an antibody-drug conjugate, etc.). The compound may contain at least one amino, imino, hydroxyl, or carboxyl group, and therefore may form a salt with the corresponding acid or base. Exemplary salts include, but are not limited to: sulfates, trifluoroacetates, citrates, acetates, oxalates, hydrochlorides, hydrobromides, hydroiodates, nitrates, hydrogen sulfates, phosphates, acid phosphates, phosphites, isonicotinates, lactates, salicylates, acid citrates, tartrates, oleates, tannates, pantothenates, hydrogen tartrate, ascorbic acid salts, formates, benzoates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, potassium salts, sodium salts, ammonium salts, calcium salts, etc.

[0192] The compounds of this invention can exist in isotopically traced or enriched forms, containing one or more atoms whose atomic weights or mass numbers differ from the atomic weights or mass numbers of the most abundant atoms found in nature. The isotopes can be radioactive or non-radioactive. Commonly used isotopes for isotopic labeling are: hydrogen isotopes: 2 H and 3 H; Carbon isotopes: 13 C and 14 C; Chlorine isotopes: 35 Cl and 37 Cl; Fluorine isotopes: 18 F; Iodine isotopes: 123 I and 125 I; Nitrogen isotopes:13 N and 15 N; oxygen isotopes: 15 O, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. Especially 2 H and 13 C, because they are easy to label and convenient to detect, are more widely used. Some heavy isotopes, such as deuterium (… 2 Substitution of H) can enhance metabolic stability and prolong half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotopically labeled compounds are generally synthesized from pre-labeled starting materials using known synthetic techniques, just like non-isotopically labeled compounds. This application includes various deuterated forms. Each available hydrogen atom bonded to a carbon atom can be independently replaced by a deuterium atom. In addition to commercially available deuterated molecular building blocks, commercially available deuterated starting materials can be used in the preparation of deuterated products, or they can be synthesized using deuteration reagents through conventional techniques. Non-limiting examples of deuteration reagents include: deuterated water, deuterated acetone, deuterated methanol, deuterated acetonitrile, deuterated borane, sodium deuterated borohydride, lithium aluminum deuteride, etc.

[0193] The term "alkyl" refers to a monovalent saturated aliphatic straight-chain or branched hydrocarbon group. "C" 1-6 "Alkyl" refers to a group that is alkyl, and the number of carbon atoms in the carbon chain is 1, 2, 3, 4, 5, or 6. This includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc.

[0194] The term "alkoxy" refers to a group in which a hydroxyl group is replaced by an alkyl group, as defined above. "C" 1-6 "Alkoxy" refers to an alkyl group that has substituted hydroxyl groups and contains 1, 2, 3, 4, 5, or 6 carbon atoms. This includes, but is not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, pentoxy, 1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, and 1-ethylpropoxy.

[0195] The term "alkathioyl" refers to a group in which a mercapto group is replaced by an alkyl group, as defined above. "C" 1-6"Alkylthio" refers to an alkyl group that replaces a mercapto group and contains 1, 2, 3, 4, 5, or 6 carbon atoms. This includes, but is not limited to, methylthio, ethylthio, propylthio, isopropylthio, butylthio, 1-methylpropylthio, 2-methylpropylthio, tert-butylthio, pentylthio, 1-methylbutylthio, 2-methylbutylthio, 3-methylbutylthio, 1,1-dimethylpropylthio, 1,2-dimethylpropylthio, 2,2-dimethylpropylthio, and 1-ethylpropylthio.

[0196] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing at least one double bond. "C" 2-6 "Alkenyl" refers to an alkenyl group composed of 2, 3, 4, 5, or 6 carbon atoms. It includes, but is not limited to, -CH=CH2, -CH=CH-CH3, -CH=CH-CH2-CH3, -CH2-CH=CH-CH3, -C(CH3)=CH-CH2-CH3, -CH=C(CH3)-CH2-CH3, -C(CH3)=CH2, -C(CH3)=CH(CH3), etc.

[0197] The term "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing at least one triple bond. "C" 2-6 "Alkyne" refers to an alkyne group composed of 2, 3, 4, 5, or 6 carbon atoms. It includes, but is not limited to, -C≡CH, -C≡C-CH3, -CH2-C≡CH, -C≡C-CH2-CH3, and -CH2-C≡C-CH3.

[0198] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0199] The term "haloalkyl" refers to a group in which one, two, three, more, or all of the hydrogen atoms on an alkyl group are replaced by a halogen. "C" 1-6 "Halogenated alkyl" refers to alkyl halogens that contain a total of 1, 2, 3, 4, 5, or 6 carbon atoms. These include, but are not limited to, -CCl3, -CHCl2, -CH2Cl, -CF3, -CHF2, -CBr3, -CHBr2, -CI3, -CHI2, -CH2I, -CH2CF3, and -CF2CF3.

[0200] The term "aryl" refers to aromatic carbocyclic systems containing 6-16 carbon atoms, including monocyclic, bicyclic, and tricyclic systems. These include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthryl, and pyreneyl.

[0201] The term "heteroatoms" refers to nitrogen, oxygen, sulfur, phosphorus, or halogens.

[0202] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic system containing a 5-14 member structure, wherein the system contains at least one heteroatom, as defined above. This includes, but is not limited to, pyridinyl, pyrazinyl, pyridazinyl, furanyl, thiopheneyl, pyrroleyl, pyrazolyl, imidazoleyl, oxazolyl, isoxazolyl, thiazolyl, or isothiazolyl. The compounds mentioned are phenyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, furanyl, thiopheneyl, pyrroloyl, pyrazolyl, imidazoleyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl, isoquinolinyl, pterinyl, purineyl, indoleyl, isoindoleyl, indazoleyl, benzofuranyl, benzothiopheneyl, benzopyridinyl, benzopyrazinyl, benzoimidazolyl, pyrrolo[2,3-b]pyridinyl, imidazo[1,2-a]pyridinyl, pyrazolo[1,5-a]pyridinyl, pyrazolo[1,5-a]pyrimidinyl, and imidazo[1,2-b]pyridazinyl.

[0203] The term "polyol" refers to a substituent containing two or more aliphatic hydroxyl groups, including but not limited to glycosyl groups and heteroatom-substituted glycosyl groups. The glycosyl groups can undergo tautomerism to form chain or cyclic structures.

[0204] The term "cycloalkyl" refers to a fully saturated carbon ring that can exist as a monocyclic, bridged, spirocyclic, or fused ring. This includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, norbornyl, bicyclo[2.2.2]octyl, adamantyl, etc.

[0205] The term "cycloalkenyl" refers to a carbon ring containing at least one double bond. Unlike aryl groups, cycloalkenyl groups are typically not fully saturated, rather than completely unsaturated. "Cyclohexenyl" refers to a six-membered carbon ring containing at least one double bond. This includes, but is not limited to, […]. wait.

[0206] The term "heterocyclic alkyl" refers to a fully saturated carbon ring containing heteroatoms that can exist as a monocyclic, bridged, spirocyclic, or fused ring. The definition of the heteroatoms is as above. These include, but are not limited to, piperidinyl, piperazinyl, homopiperazinyl, morpholinyl, tetrahydrofuranyl, pyrrolidinyl, imidazoalkyl, azirropropyl, oxacyclopropyl, thioazirropropyl, azirrobutyl, oxacyclobutyl, thioazirrobutyl, oxacyclohexyl, thiomorpholinyl, oxathiohexyl, and thiazoalkyl.

[0207] The term "heterocyclic alkenyl" refers to a carbon ring containing at least one double bond, containing one, two, or three heteroatoms (one, two, or three of N, O, and S), and having a specified number of carbon atoms (e.g., 1, 2, or 3). Unlike heteroaryl groups, heterocyclic alkenyl groups are typically not fully saturated, rather than completely unsaturated.

[0208] The term "alkylene" refers to a saturated divalent hydrocarbon group obtained by removing two hydrogen atoms from a saturated straight-chain or branched hydrocarbon group; that is, one hydrogen atom in the alkyl group is replaced, as defined above. Examples of alkylene groups include methylene (-CH2-), ethylene {including -CH2CH2- or -CH(CH3)-}, isopropylene {including -CH(CH3)CH2- or -C(CH3)2-}, etc.

[0209] The term "alkenyl" refers to a substituent formed by eliminating two hydrogen atoms from a straight-chain or branched alkene containing one or more carbon-carbon double bonds and no carbon-carbon triple bonds, which has a specific number of carbon atoms. The carbon-carbon double bonds can be located anywhere within the alkenyl group, and the two eliminated hydrogen atoms can be on the same carbon atom or on different carbon atoms (e.g., the two eliminated hydrogen atoms are on the carbon atoms at the two ends). Therefore, C2 alkenyl groups (i.e., vinylidenes) include, but are not limited to, -CH=CH-, and C3 alkenyl groups include, but are not limited to, -CH2-CH=CH-. And -C(CH3)=CH-, C4 imidenes include but are not limited to -CH2-CH=CH-CH2-, -CH2=CH-CH2-CH2- and -CH2-CH-CH2=CH2-.

[0210] The term "ethynyl" refers to a substituent formed by eliminating two hydrogen atoms from a straight-chain or branched alkene containing one or more carbon-carbon triple bonds and a specific number of carbon atoms. The carbon-carbon triple bond can be located anywhere within the ethynyl group, and the two eliminated hydrogen atoms can be on the same carbon atom or on different carbon atoms (e.g., the two eliminated hydrogen atoms are on the two end carbon atoms). Therefore, C2 ethynyl groups (i.e., ethynyl groups) include, but are not limited to, […]. C3 ethynyl group includes, but is not limited to, C4 ethynyl group includes, but is not limited to,

[0211] The terms “cycloalkylene,” “cycloalkenylene,” “heterocycloalkylene,” “arylene,” “heteroarylene,” and “heterocycloalkenylene” used in this invention refer to the further substitution of one hydrogen atom of cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, heteroaryl, or heterocycloalkenyl, as defined above.

[0212] The term "pharmaceutical excipients" refers to all substances contained in a pharmaceutical preparation other than the active pharmaceutical ingredient.

[0213] The term “treatment” refers to a therapeutic approach. When a specific condition is involved, treatment means: (1) alleviating one or more biological manifestations of the disease or condition; (2) interfering with (a) one or more points in a biological cascade that causes or precipitates the condition or (b) one or more biological manifestations of the condition; (3) improving one or more symptoms, effects or side effects associated with the condition, or one or more symptoms, effects or side effects associated with the condition or its treatment; or (4) slowing the development of the disease or one or more biological manifestations of the condition.

[0214] The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0215] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0216] The reagents and raw materials used in this invention are all commercially available.

[0217] The positive and progressive effects of this invention are as follows: the camptothecin compounds provided by this invention have one or more of the following advantages:

[0218] (1) It has good inhibitory activity against tumor cells;

[0219] (2) Good liver microsomal stability;

[0220] (3) It has good inhibitory activity against topoisomerase I.

[0221] The camptothecin-based antibody-drug conjugates provided by this invention have one or more of the following advantages:

[0222] (1) It has good inhibitory activity against tumor cells;

[0223] (2) Good release efficiency;

[0224] (3) The plasma has good stability;

[0225] (4) It has the bystander effect;

[0226] (5) Good in vivo antitumor inhibitory activity. Detailed Implementation

[0227] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0228] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (MeOD-d4) as solvents. LC-MS analysis was performed using high-performance liquid chromatography (HPLC) with a diode array (DAD) UV detector and an Xbridge C18 2.1*50mm 5µm column. Detection methods included diode array (DAD) and evaporative light scattering (ELSD), with positive / negative ion mode scanning and a mass scan range of 100–1000. A rapid column purification system, Isolara One (Biotage), was used. For normal column chromatography, 300–400 mesh silica gel was typically used as the support.

[0229] For ease of expression, some technical terms are abbreviated as follows: Pd(dppf)Cl2, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride; Dioxane, 1,4-dioxane; HATU, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; DIPEA, diisopropylethylamine; TFA, trifluoroacetic acid; Boc, tert-butyloxycarbonyl; Ac, acetyl; TEA, triethylamine; DCM, dichloromethane; Bn, benzyl; DMF, N,N-dimethylformamide; BPD, pinacol diboronate; cataCXium A-Pd-G2, chloro[(n-butyldi(1-adamantyl)phosphine)-2-(2-aminobiphenyl)]palladium(II); Bpin, 4,4,5,5-tetramethyl-1,3,2-dioxoboronyl; HOPO, 2-hydroxypyridine-N-oxide; EDCI, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride; Su, succinimide; EDC Br, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrobromide; Trastuzumab, trastuzumab.

[0230] Payload synthesis:

[0231] Example 1. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-(4-hydroxyphenyl)-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P1)

[0232] The synthesis of compound 8 can be found in Chinese Patent CN 115551552 A (Publication No.), and the same applies below.

[0233] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.01 mmol), sodium carbonate (57.69 mg, 0.54 mmol), and compound P-1-1 (22.52 mg, 0.16 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40 mm×5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 min) to obtain a white solid compound P1 (16.0 mg, yield 29.6%). LCMS:(ESI,m / z):473.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.77(s,1H),8.08(s,1H),7.94(d,J=10.89Hz,1H),7.34(s,1H),7.14(d,J=8.29Hz,2H),6.98(d, J=8.41Hz,2H),6.52(s,1H),5.40(s,2H),5.16(s,2H),2.19(d,J=2.10Hz,3H),1.78-1.93(m,2H),0.88(t,J=7.30Hz,3H).

[0234] Example 2. (19S)-8-(4-aminophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P2)

[0235] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.01 mmol), sodium carbonate (57.7 mg, 0.54 mmol), and compound P-2-1 (22.5 mg, 0.16 mmol) were added to a mixed solution of compound 8 (50 mg, 0.11 mmol) in dioxane (2 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40 mm×5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 min) to obtain a yellow solid compound P2 (18.8 mg, yield 33.7%).

[0236] LCMS:(ESI,m / z):472.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.15(s,1H),7.90(d,J=10.64Hz,1H),7.33(s,1H),6.97(d,J=8.17Hz,2H),6.76(d,J=8.29Hz,2H),6.51 (s,1H),5.41(s,2H),5.37(s,2H),5.17(s,2H),2.21(d,J=2.10Hz,3H),1.86(tt,J=14.18,7.10Hz,2H),0.88(t,J=7.42Hz,3H).

[0237] Example 3. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-(3-hydroxyphenyl)-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P5)

[0238] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.01 mmol), sodium carbonate (57.7 mg, 0.54 mmol), and compound P-5-1 (22.52 mg, 0.16 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (1.5 mL) and water (0.3 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 15%-45% B, retention 8 min) to give a yellow solid compound P5 (20.5 mg, yield 38.7%). LCMS: (ESI, m / z): 473.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.78(td,J=9.12,1.55Hz,1H),8.04(s,1H),7.95(d,J=10.76Hz,1H),7.41(t,J=7.79Hz,1H),7.34(s,1H),6.90-6.99(m,1 H),6.73(dt,J=7.45,1.28Hz,1H),6.67-6.70(m,1H),5.40(s,2H),5.15( s,2H),2.17(d,J=2.10Hz,3H),1.77-1.93(m,2H),0.87(t,J=7.30Hz,3H).

[0239] Example 4. (19S)-8-(3-aminophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P6)

[0240] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.01 mmol), sodium carbonate (57.7 mg, 0.54 mmol), and compound P-6-1 (22.4 mg, 0.16 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (1.50 mL) and water (0.30 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 15%-45% B, retention 8 min) to give a yellow solid compound P6 (16.1 mg, yield 24%). LCMS: (ESI, m / z): 472.1 [M+H] + 1 H NMR (400MHz, DMSO-d6) δ8.06 (s, 1H), 7.96 (d, J = 10.88Hz, 1H), 7.28-7.39 (m, 2H), 6.87 (br d,J=7.63Hz,1H),6.50-6.70(m,2H),5.41(s,2H),5.17(s,2H),2.19(d,J=2.00Hz,3H),1.77-1.98(m,2H),0.88(t,J=7.32Hz,3H).

[0241] Example 5.2-Amino-N-{4-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenylacetamide (P3)

[0242] Step 1: Synthesis of tert-butyl N-({[4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)phenyl]aminocarbonyl}methyl)aminomethyl ester (P-3-2)

[0243] At room temperature, N,N-diisopropylethylamine (294 mg, 2.28 mmol), 2-{[(tert-butoxy)carbonyl]amino}acetic acid (399 mg, 2.28 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (867 mg, 2.28 mmol) were sequentially added to a 10 mL solution of compound P-2-1 (500 mg, 2.28 mmol). The reaction mixture was stirred at 25 °C for 1 hour. The reaction solution was filtered, and the filtrate was separated by high performance liquid preparative chromatography (column type: WePure Biotech XP tC18(w) 250×70mm 10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 35%-70% B, retention 20 min) to obtain a yellow solid compound P-3-2 (150 mg, yield 17.4%).

[0244] Step 2: 2-Amino-N-{4-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}acetate (P3)

[0245] At room temperature, a solution of sodium carbonate (173 mg, 1.63 mmol) in water (0.6 mL) and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (26.6 mg, 0.032 mmol) were added to a solution of compound 8 (150 mg, 0.32 mmol) and compound P-3-2 (147 mg, 0.39 mmol) in dioxane (2.4 mL). The reaction mixture was reacted at 100 °C for 1 hour under nitrogen protection. After the reaction was complete, 10 mL of trifluoroacetic acid was slowly added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. The reaction solution was filtered, and the filtrate was separated by high-performance liquid chromatography (HPLC) using a preparative column (Phenomenex Luna C18(w) 75×30mm 3µm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 10%-50% B, retention time 8 min) to obtain a yellow solid compound P3 (20.3 mg, yield 9.67%). LCMS (ESI, m / z): 529.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.70(s,1H),8.19(br s,3H),7.93-8.05(m,2H),7.83(d,J=8.63Hz,2H),7.29-7.40(m,3H),6.52(br s,1H),5.40(s,2H),5.14(s,2H),3.87(br s,2H),2.18(d,J=2.13Hz,3H),1.78-1.95(m,2H),0.88(t,J=7.32Hz,3H).

[0246] Example 6. N-{4-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}-2-hydroxyacetamide (P4)

[0247] Step 1: Synthesis of {[4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)phenyl]aminocarbonyl}methyl acetate (P-4-2)

[0248] At room temperature, 2-chloro-2-oxoylidelethyl acetate (467 mg, 3.42 mmol) and triethylamine (346 mg, 3.42 mmol) were sequentially added to a solution of compound P-2-1 (500 mg, 2.28 mmol) in dichloromethane (10.0 mL). The reaction mixture was stirred at 25 °C for 1 hour and then filtered. The filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18(w) 250 × 70 mm 10 μm; mobile phase [water (10 mM ammonium bicarbonate)-acetonitrile]; mobile phase: 30%-65% B, retention 20 min) to obtain a yellow solid compound P-4-2 (200 mg, yield 27.4%). LCMS (ESI, m / z): 320.2 [M+H] + .

[0249] Step 2: N-{4-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 ,9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}-2-hydroxyacetamide (P4)

[0250] At room temperature, a solution of sodium carbonate (173 mg, 1.63 mmol) in water (0.6 mL) and [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (26.6 mg, 0.032 mmol) were added sequentially to a solution of compound 8 (150 mg, 0.32 mmol) and compound P-4-2 (135 mg, 0.42 mmol) in dioxane (2.4 mL). The reaction mixture was reacted at 100 °C for 1 hour under nitrogen protection. After the reaction was completed, the reaction mixture was filtered at room temperature. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) using a Phenomenex Luna C18(w) 75 × 30 mm 3 μm column; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 10%-40% B, retention 8 min) to obtain a yellow solid compound P4 (16.7 mg, yield 7.95%). LCMS(ESI,m / z):530.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.93(s,1H),8.05(s,1H),7.93-8.00(m,3H),7.34(s,1H),7.29(d,J=8.51Hz,2H),6.51(br s,1H),5.40(s,2H),5.15(s,2H),4.06(s,2H),2.19(d,J=2.00Hz,3H),1.77-1.95(m,2H),0.88(t,J=7.32Hz,3H).

[0251] Example 7.2-Amino-N-{3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}acetate (P7)

[0252] Step 1: Synthesis of tert-butyl N-({[3-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)phenyl]aminocarbonyl}methyl)aminomethyl ester (P-7-2)

[0253] 2-(tert-Butoxycarbonylamino)acetic acid (599 mg, 3.42 mmol), N,N-diisopropylethylamine (4.87 mg, 6.85 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.30 g, 3.42 mmol) were added to 10 mL of dichloromethane containing compound P-7-1 (0.5 g, 2.28 mmol). The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 250×70mm×10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 30%-65% B, retention 20 min) to obtain a yellow solid compound P-7-2 (150 mg, yield 17.47%). LCMS: (ESI, m / z): 321.2 [M+H-56] + .

[0254] Step 2: 2-Amino-N-{3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}acetate (P7)

[0255] Under a nitrogen atmosphere, [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (26.6 mg, 0.03 mmol), sodium carbonate (173 mg, 1.63 mmol), and compound P-7-2 (147 mg, 0.39 mmol) were added to a mixed solution of compound 8 (150 mg, 0.33 mmol) in dioxane (6 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, trifluoroacetic acid (1.50 mL) was added to the reaction solution. The reaction solution was stirred at 25 °C for another 2 hours. The reaction solution was filtered and concentrated to obtain a yellow oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 15%-45% B, retention 8 min) to give a yellow solid compound P7 (20.61 mg, yield 11.65%). LCMS: (ESI, m / z): 529.0 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ10.69(s,1H),8.14(br s,3H),7.95-8.03(m,2H),7.76(br d,J=8.29Hz,1H),7.58-7.67(m,2H),7.34(d,J=1.11Hz,1H),7.10(dd,J=7.36,3.53Hz,1H),6.53(br s,1H),5.40(s,2H),5.04-5.21(m,2H),3.82(br s,2H),2.16(d,J=1.86Hz,3H),1.86(dt,J=13.83,6.88Hz,2H),0.88(t,J=7.36Hz,3H).

[0256] Example 8 N-{3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}-2-hydroxyacetamide (P8)

[0257] Step 1: Synthesis of {[3-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)phenyl]aminocarbonyl}methyl acetate (P-8-2)

[0258] Compound P-7-1 (0.50 g, 2.28 mmol) was dissolved in dichloromethane (10 mL), and 2-chloro-2-oxoylide ethyl acetate (311 mg, 2.28 mmol) and triethylamine (231 mg, 2.28 mmol) were added separately. The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a yellow oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 250×70 mm×10 μm; mobile phase [water (10 mM ammonium bicarbonate)-acetonitrile]; mobile phase: 30%-65% B, retention 20 min) to obtain a yellow solid compound P-8-2 (150 mg, yield 17.47%). LCMS: (ESI, m / z): 320.2 [M+H] + .

[0259] Step 2: N-{3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}-2-hydroxyacetamide (P8)

[0260] Under a nitrogen atmosphere, [1,1-bis(diphenylphosphine)ferrocene]palladium dichloride (26.7 mg, 0.033 mmol), sodium carbonate (173 mg, 1.63 mmol), and compound P-8-2 (125 mg, 0.392 mmol) were added to a mixed solution of compound 8 (150 mg, 0.33 mmol) in dioxane (3 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered at room temperature, and the filtrate was concentrated to obtain a yellow oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna 100×40 mm×5 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 min) to obtain a yellow solid compound P8 (20.42 mg, yield 11.36%). LCMS:(ESI,m / z):530.0[M+H] + . 1H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 8.03 (s, 1H), 7.98 (d, J = 10.89Hz, 1H), 7.89 (br d,J=8.17Hz,1H),7.73(s,1H),7.55(t,J=7.92Hz,1H),7.34(s,1H),7.03(d,J=7.42Hz,1H),6.53(br s,1H),5.40(s,2H),5.15(s,2H),4.01(s,2H),2.17(d,J=1.48Hz,3H),1.79-1.95(m,2H),0.88(t,J=7.30Hz,3H).

[0261] Example 9 (19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-8-(6-oxoylide-1,6-dihydropyridin-3-yl)-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P11)

[0262] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (12.45 mg, 0.15 mmol), potassium carbonate (105.32 mg, 0.76 mmol), and compound P-11-1 (46.32 mg, 0.3 mmol) were added to a mixed solution of compound 8 (70.0 mg, 0.15 mmol) in dioxane (2.8 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100×30mm×5um; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 20%-50% B, retention 8 min) to obtain a yellow solid compound P11 (16.20 mg, yield 20.8%). LCMS: (ESI, m / z): 487.1 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ7.95-8.02(m,2H),7.54-7.61(m,1H),7.48-7.54(m,1H),7.34(s,1H),7.27(s,1H),7.21(br d,J=7.38Hz,1H),6.34-6.71(m,1H),5.40(s,2H),5.15(s,2H),4.63(s,2H),2.17(s,3H),1.77-1.96(m,2H),0.88(br t,J=7.25Hz,3H).

[0263] Example 10. (19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-8-(6-oxoylide-1,6-dihydropyridin-3-yl)-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P12)

[0264] Step 1: Synthesis of (S)-10-(6-benzyloxy)pyridin-3-yl)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-1,12-dihydro-14H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-3,14(4H)-dione (P-12-1)

[0265] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.11 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and (6-(benzyloxy)pyridin-3-yl)boronic acid (49.87 mg, 0.22 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2.0 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Welch Ultimate 120×30mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 45%-80% B, retention 8 min) to give a yellow solid compound P-12-1 (30.0 mg, yield 48.8%). LCMS: (ESI, m / z): 564.3 [M+H] + .1 H NMR (400MHz, DMSO-d6) δ8.19(d,J=1.13Hz,1H),8.12(s,1H),8.01(d,J=10.76Hz,1H),7.72-7.81(m,1H),7.54(br d,J=7.13Hz,2H),7.29-7.47(m,4H),7.14(d,J=8.50Hz,1H),6.24-6.79(m,1H),5. 36-5.51(m,4H),5.16(s,2H),2.21(s,3H),1.86(dt,J=14.13,6.94Hz,2H),0.88(br t,J=7.32Hz,3H).

[0266] Step 2: (19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-8-(6-oxoylide-1,6-dihydropyridin-3-yl)-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P12)

[0267] Boron tribromide (1 mol / L tetrahydrofuran solution, 212.93 μL) was added dropwise to a dichloromethane (0.3 mL) solution of compound P-12-1 (30.0 mg, 0.05 mmol) under a nitrogen atmosphere. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was quenched with water (0.5 mL) and filtered, and concentrated to obtain crude lutein solid. The crude product was separated by high performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30 mm×10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 10%-50% B, retention 8 min) to obtain a yellow solid compound P12 (10.5 mg, yield 40.95%).

[0268] LCMS:(ESI,m / z):474.2[M+H] + . 1H NMR(400MHz,DMSO-d6)δ12.01(br s,1H),8.35(s,1H),7.98(d,J=10.76Hz,1H),7.39-7.55(m,2H),7.34(s,1H),6.48-6.59(m,2H),5.42 (s,2H),5.19(s,2H),2.28(d,J=1.88Hz,3H),1.86(tt,J=14.09,6.86Hz,2H),0.88(t,J=7.32Hz,3H).

[0269] Example 11. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-[1-(2-hydroxyethyl)-6-oxoylide-1,6-dihydropyridin-3-yl]-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P13)

[0270] Step 1: Synthesis of 5-bromo-1-(2-hydroxyethyl)-1,2-dihydropyridin-2-one (P-13-2)

[0271] Potassium carbonate (1.59 g, 11.49 mmol) was added to a solution of 2-bromoethane-1-ol (1.44 g, 11.49 mmol) and 5-bromo-1,2-dihydropyridin-2-one (1 g, 5.75 mmol) in N,N-dimethylformamide (20 mL) at room temperature. The reaction was stirred at 25 °C for 12 hours, diluted with ethyl acetate (50 mL), and filtered. The filtrate was concentrated to give a yellow oily compound P-13-2 (1 g, 79.8% yield). LCMS: (ESI, m / z): 217.8 / 219.8 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.87(br s,1H),7.46-7.57(m,1H),6.36(br d,J=9.66Hz,1H),4.91(br t,J=5.14Hz,1H),3.91(br t,J=5.01Hz,2H),3.60(br d,J=5.14Hz,2H).

[0272] Step 2: Synthesis of 1-(2-hydroxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)-1,2-dihydropyridin-2-one (P-13-3)

[0273] Dioxane (5 mL) was added to a mixture of 5-bromo-1-(2-hydroxyethyl)-1,2-dihydropyridin-2-one (0.5 g, 2.29 mmol), pinacol diboronate (1.75 g, 6.88 mmol), potassium acetate (450.10 mg, 4.59 mmol), and 1,1-bis(diphenylphosphine)ferrocene palladium chloride (167.79 mg, 229.31 μmol) at room temperature, and the mixture was bubbled under nitrogen atmosphere at 95 °C for 1 hour. After the reaction was complete, the mixture was diluted with ethyl acetate (50 mL) and filtered through diatomaceous earth. The filtrate was concentrated to give a brown oily compound. The crude product was separated by column chromatography (eluent: ethyl acetate) to give a yellow solid compound P-13-3 (0.5 g, yield 82.2%). LCMS: (ESI, m / z): 266.1 [M+H] + .

[0274] Step 3: (19S)-19-ethyl-6-fluoro-19-hydroxy-8-[1-(2-hydroxyethyl)-6-oxoylide-1,6-dihydropyridin-3-yl]-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P13)

[0275] At room temperature, (19S)-8-bromo-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20A mixture of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (40 mg, 87.10 μmol), 1-(2-hydroxyethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)-1,2-dihydropyridin-2-one (57.73 mg, 217.74 μmol), 1,1-di(tert-butylphosphine)ferrocene palladium chloride (5.68 mg, 8.71 μmol) and sodium bicarbonate (21.95 mg, 261.29 μmol) was added to dioxane (1.6 mL) and water (0.4 mL) and bubbled with nitrogen. The reaction was carried out at 100 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated by high-performance liquid chromatography (HPLC) using a WePure Biotech XP tC18 column (100×30mm×10um; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 15%-45% B, retention time: 8 min) to obtain a white solid compound P13 (15.6 mg, yield: 34.6%). LCMS (ESI, m / z): 518.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.41(s,1H),7.98(d,J=10.76Hz,1H),7.69(d,J=2.50Hz,1H),7.44(dd ,J=9.26,2.50Hz,1H),7.34(s,1H),6.45-6.63(m,2H),5.42(s,2H),5.09-5.26(m,2H),4.04(br t,J=5.13Hz,2H),3.72(br t,J=5.38Hz,2H),2.31(d,J=1.88Hz,3H),1.87(tquin,J=14.07,14.07,7.06,7.06,7.06,7.06Hz,2H),0.88(t,J=7.25Hz,3H).

[0276] Example 12. (19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-8-(2-oxoylide-1,2-dihydropyridin-4-yl)-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P14)

[0277] Under a nitrogen atmosphere, chloro[(n-butyldi(1-adamantyl)phosphine)-2-(2-aminobiphenyl)]palladium(II) (7.93 mg, 0.01 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and compound P-14-1 (45.3 mg, 0.33 mmol) were added to a mixed solution of compound 8 (50 mg, 0.11 mmol) in dioxane (2 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30mm×10um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 15%-50% B, retention 8 min) to obtain a yellow solid compound P14 (6 mg, yield 10.59%). LCMS: (ESI, m / z): 474.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.28(s,1H),7.99(d,J=10.79Hz,1H),7.61(d,J=6.53Hz,1H),7.34(s,1H),6.31(s,1H),6.15(br d,J=6.78Hz,1H),5.41(s,2H),5.12-5.27(m,2H),2.27(d,J=1.76Hz,3H),1.86(dt,J=14.40,7.17Hz,2H),0.88(t,J=7.28Hz,3H).

[0278] Example 13. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-[1-(2-hydroxyethyl)-2-oxoylide-1,2-dihydropyridin-4-yl]-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P15)

[0279] Step 1: Synthesis of 4-bromo-1-(2-hydroxyethyl)-1,2-dihydropyridin-2-one (P-15-2)

[0280] Potassium carbonate (1.59 g, 11.49 mmol) was added to a solution of 2-bromoethane-1-ol (1.44 g, 11.49 mmol) and 4-bromo-1,2-dihydropyridin-2-one (1 g, 5.75 mmol) in N,N-dimethylformamide (20 mL) at room temperature. The reaction was stirred at 25 °C for 12 hours, diluted with ethyl acetate (50 mL), and filtered. The filtrate was concentrated to give a yellow oily compound P-15-2 (0.9 g, 71.8% yield). LCMS: (ESI, m / z): 218.0 / 220.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.57(d,J=7.21Hz,1H),6.69(d,J=2.08Hz,1H),6.43(dd,J=7.2 1,2.20Hz,1H),4.89(t,J=5.44Hz,1H),3.90(t,J=5.38Hz,2H),3.59(q,J=5.42Hz,2H).

[0281] Step 2: Synthesis of 1-(2-hydroxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)-1,2-dihydropyridin-2-one (P-15-3)

[0282] Dioxane (5 mL) was added to a mixture of 4-bromo-1-(2-hydroxyethyl)-1,2-dihydropyridin-2-one (0.5 g, 2.29 mmol), pinacol diboronate (1.75 g, 6.88 mmol), potassium acetate (450.10 mg, 4.59 mmol), and 1,1-bis(diphenylphosphine)ferrocene palladium chloride (167.79 mg, 229.31 μmol) at room temperature, and the mixture was bubbled under nitrogen atmosphere at 95 °C for 1 hour. After the reaction was complete, the mixture was diluted with ethyl acetate (50 mL) and filtered through diatomaceous earth. The filtrate was concentrated to give a brown oily compound. The crude product was separated by column chromatography (eluent: ethyl acetate) to give a yellow solid compound P-15-3 (0.4 g, yield 65.7%). LCMS: (ESI, m / z): 266.1 [M+H] + .

[0283] Step 3: (19S)-19-ethyl-6-fluoro-19-hydroxy-8-[1-(2-hydroxyethyl)-2-oxoylide-1,2-dihydropyridin-4-yl]-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 ,20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P15)

[0284] At room temperature, (19S)-8-bromo-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 A mixture of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (50 mg, 108.87 μmol), 1-(2-hydroxyethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)-1,2-dihydropyridin-2-one (86.59 mg, 326.61 μmol), 1,1-bis(diphenylphosphine)ferrocene palladium chloride (7.97 mg, 10.89 μmol) and potassium carbonate (45.14 mg, 326.61 μmol) was mixed with dioxane (1.6 mL) and water (0.4 mL) and bubbled with nitrogen. The mixture was reacted at 100 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered through a filter head, and the filtrate was separated by high-performance liquid chromatography (HPLC) using a preparative column (column type: WePure Biotech XP tC18 100×30mm×10um; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 20%-50% B, retention time: 8 min) to obtain a yellow solid compound P15 (14.7 mg, yield: 26.0%). LCMS: (ESI, m / z): 518.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.27(br s,1H),8.00(br d,J=10.63Hz,1H),7.83(br d,J=6.25Hz,1H),7.34(s,1H),6.52(br s,1H),6.38(br s,1H),6.19(br d,J=4.88Hz,1H),5.41(br s,2H),5.14-5.27(m,2H),4.91-5.06(m,1H),4.02-4.13(m,2H),3.75(br s,2H),2.28(br s,3H),1.79-1.93(m,2H),0.88(br t,J=6.75Hz,3H).

[0285] Example 14. (19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-8-(1H-pyrazol-4-yl)-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P16)

[0286] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (12.14 mg, 0.01 mmol), potassium carbonate (105.33 mg, 0.76 mmol), and compound P-16-1 (118.3 mg, 0.61 mmol) were added to a mixed solution of compound 8 (70.0 mg, 0.15 mmol) in dioxane (2.8 mL) and water (0.4 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100×30mm×5um; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 20%-50% B, retention 8 min) to obtain a yellow solid compound P16 (12.2 mg, yield 16.42%). LCMS: (ESI, m / z): 447.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.35(s,1H),7.91(d,J=10.92Hz,1H),7.85(s,2H),7.33(s,1H),5.41(s, 2H), 5.16 (s, 2H), 2.28 (d, J = 2.13Hz, 3H), 1.86 (dt, J = 14.43, 6.96Hz, 2H), 0.87 (t, J = 7.34Hz, 3H).

[0287] Example 15. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-[1-(2-hydroxyethyl)-1H-pyrazol-4-yl]-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P17)

[0288] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (12.45 mg, 0.02 mmol), potassium carbonate (105.3 mg, 0.76 mmol), and compound P-17-1 (22.4 mg, 0.16 mmol) were added to a mixed solution of compound 8 (70.0 mg, 0.15 mmol) in dioxane (2.50 mL) and water (0.50 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 75×30mm×3um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 15%-45% B, retention 8 min) to obtain a yellow solid compound P17 (16.1 mg, yield 21.05%). LCMS: (ESI, m / z): 491.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.41(s,1H),7.97(s,1H),7.92(d,J=10.88Hz,1H),7.63(s,1H),7.33(s,1H),6.52(s,1H),5.41(s,2H),5.19(s,2H) ,5.00(t,J=5.32Hz,1H),4.30(t,J=5.63Hz,2H),3.86(q,J=5.59Hz,2H),2.31(d,J=2.00Hz,3H),1.78-1.95(m,2H),0.88(t,J=7.32Hz,3H).

[0289] Example 16. (19S)-8-(3-amino-4-fluorophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P18)

[0290] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (7.97 mg, 0.0108 mmol), potassium carbonate (45.14 mg, 0.326 mmol), and compound P-18-1 (38.72 mg, 0.163 mmol) were added to a mixed solution of compound 8 (0.05 g, 0.108 mmol) in dioxane (1.6 mL) and water (0.4 mL). After the reaction apparatus was purged three times, the reaction was carried out at 90 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (Phenomenex luna C18 100×30mm×5um column; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 min) to obtain a yellow solid compound P18 (17.80 mg, yield 33.4%). LCMS (ESI, m / z): 490.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.08(s,1H),7.94(d,J=10.88Hz,1H),7.33(s,1H),7.20(dd,J=11.57,8.19Hz,1H),6.66(br d,J=8.50Hz,1H),6.52(s,1H),6.44(ddd,J=6.00,4.19,2.06Hz,1H),5.28-5.46(m,4H),5.16(s,2H),2.1 8(d,J=1.88Hz,3H), 1.86(tquin,J=14.04,14.04,7.08,7.08,7.08,7.08Hz,2H), 0.88(t,J=7.32Hz,3H).

[0291] Example 17. (19S)-8-(3-amino-5-methoxyphenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P19)

[0292] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15.93 mg, 21.77 μmol), potassium carbonate (30.09 mg, 217.74 μmol), and compound P-19-1 (54.24 mg, 217.74 μmol) were added to a mixed solution of compound 8 (50 mg, 108.87 μmol) in dioxane (0.8 mL) and water (0.2 mL). After the reaction apparatus was purged three times, the reaction was carried out at 80 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30×10um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 25%-55% B, retention 8 min) to obtain a yellow solid compound P19 (9.0 mg, yield 16.48%). LCMS: (ESI, m / z): 502.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.12 (s, 1H) 7.94 (d, J = 10.76Hz, 1H) 7.34 (s, 1H) 6.39 (br s, 1H) 6.12 (br d,J=10.63Hz,2H)5.41(s,2H)5.05-5.29(m,2H)3.73(d,J=1.13Hz,3H)2.21(d,J=2.0 0Hz, 3H) 1.86 (dquin, J = 14.20, 6.99, 6.99, 6.99, 6.99Hz, 2H) 0.88 (t, J = 7.32Hz, 3H).

[0293] Example 18. (19S)-8-(3-amino-5-methylphenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P20)

[0294] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.11 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and compound P-20-1 (50.76 mg, 0.22 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2.0 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100×30mm×5um; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 10%-40% B, retention 8 min) to give a yellow solid compound P20 (16.15 mg, yield 28.1%). LCMS: (ESI, m / z): 486.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.04 (s, 1H), 7.99 (d, J = 10.92Hz, 1H), 7.34 (s, 1H), 7.11 (br s,1H),6.82-6.97(m,2H),5.41(s,2H),5.06-5.26(m,2H),2.40(s,3H),2.1 8(d,J=2.01Hz,3H),1.86(dt,J=13.99,6.93Hz,2H),0.88(t,J=7.28Hz,3H).

[0295] Example 19.3-Amino-5-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]benzonitrile (P21)

[0296] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.11 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and compound P-21-1 (53.1 mg, 0.11 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2.0 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Welch Ultimate 120×30 mm×5 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 min) to obtain a yellow solid compound P21 (16.20 mg, yield 29.4%). LCMS:(ESI,m / z):497.1[M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.07(s,1H),7.99(d,J=10.88Hz,1H),7.34(s,1H),7.06(s,1H),6.86(s,1H),6.75(s,1H),6.35-6.66(m,1H), 5.41(s,2H),5.10-5.25(m,2H),2.18(d,J=1.75Hz,3H),1.86(dquin,J=14.16,7.00,7.00,7.00,7.00Hz,2H),0.88(t,J=7.32Hz,3H).

[0297] Example 20. (19S)-8-[3-amino-4-(trifluoromethyl)phenyl]-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P22)

[0298] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15.93 mg, 21.77 μmol), potassium carbonate (30.09 mg, 217.74 μmol), and compound P-22-1 (62.51 mg, 217.74 μmol) were added to a mixed solution of compound 8 (50 mg, 108.87 μmol) in dioxane (0.8 mL) and water (0.2 mL). After the reaction apparatus was purged three times, the reaction was carried out at 80 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Welch Ultimate C18 120×30mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 35%-70% B, retention 8 min) to obtain a yellow solid compound P22 (10.1 mg, yield 16.33%). LCMS: (ESI, m / z): 540.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.09(s,1H)7.99(d,J=10.88Hz,1H)7.56(d,J=8.00Hz,1H)7.34(s,1H)6.75(s,1H)6.48-6. 61(m,2H)5.68-6.01(m,2H)5.41(s,2H)5.17(s,2H)2.19(d,J=2.00Hz,3H)1.80-1.93(m,2H)0.88(t,J=7.32Hz,3H).

[0299] Example 21. (19S)-8-(3-amino-2-fluorophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P23)

[0300] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15.93 mg, 21.77 μmol), potassium carbonate (30.09 mg, 217.74 μmol), and compound P-23-1 (51.62 mg, 217.74 μmol) were added to a mixed solution of compound 8 (50 mg, 108.87 μmol) in dioxane (0.8 mL) and water (0.2 mL). After the reaction apparatus was purged three times, the reaction was carried out at 80 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100×30mm×5um; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 20%-50% B, retention 8 min) to obtain a yellow solid compound P23 (9.3 mg, yield 17.0%). LCMS: (ESI, m / z): 490.0 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.06(s,1H)8.01(d,J=10.76Hz,1H)7.35(s,1H)7.06-7.14(m,1H)6.93-7.01(m,1H )6.39-6.48(m,1H)5.41(s,2H)5.19(s,2H)2.20(d,J=1.75Hz,3H)1.78-1.94(m,2H)0.88(t,J=7.32Hz,3H).

[0301] Example 22. (19S)-8-(3-amino-2,4-difluorophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P24)

[0302] Under a nitrogen atmosphere, 1,1-bis(tert-butylphosphine)ferrocene palladium chloride (7.10 mg, 0.0108 mmol), potassium carbonate (45.14 mg, 0.326 mmol), and compound P-24-1 (41.65 mg, 0.163 mmol) were added to a mixed solution of compound 8 (0.05 g, 0.108 mmol) in dioxane (1.0 mL) and water (0.25 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (Phenomenex Luna C18 100×30mm×5um column; mobile phase [water (0.04% hydrochloric acid)-acetonitrile]; mobile phase: 15%-45% B, retention 8 min) to obtain a yellow solid compound P24 (12.2 mg, yield 20.6%). LCMS: (ESI, m / z): 508.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.97-8.12(m,2H),7.35(d,J=2.88Hz,1H),7.15(br t,J=8.57Hz,1H),6.43-6.55(m,2H),5.36-5.53(m,4H),5.17(br s,2H),2.20(br s,3H),1.78-1.95(m,2H),0.83-0.93(m,3H).

[0303] Example 23. (19S)-8-(5-aminopyridin-3-yl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P25)

[0304] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (8.89 mg, 0.11 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and compound P-25-1 (24.0 mg, 0.11 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2.0 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Welch Ultimate 120×30 mm×5 μm; mobile phase [water (0.4% hydrochloric acid)-acetonitrile]; mobile phase: 10%-40% B, retention 8 min) to obtain a yellow solid compound P25 (6.10 mg, yield 10.9%). LCMS:(ESI,m / z):473.1[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=1.88Hz,1H),8.20(s,1H),8.09-8.17(m,1H),8.03-8.09(m,2H),7.90-8.01(m,1H),7.48(s,1H),7.35 (s,1H),5.41(s,2H),5.15(s,2H),2.22(d,J=1.63Hz,3H),1.87(dquin,J=13.88,7.01,7.01,7.01,7.01Hz,2H),0.88(t,J=7.25Hz,3H).

[0305] Example 24. (19S)-8-(5-amino-2-fluorophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P26)

[0306] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (7.97 mg, 0.0108 mmol), potassium carbonate (45.14 mg, 0.326 mmol), and compound P-26-1 (38.72 mg, 0.163 mmol) were added to a mixed solution of compound 8 (0.05 g, 0.108 mmol) in dioxane (1.6 mL) and water (0.4 mL). After the reaction apparatus was purged three times, the reaction was carried out at 90 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Ultimate C18 120×30mm×5um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 20%-50% B, retention 8 min) to give a yellow solid compound P26 (17.10 mg, yield 32.0%). LCMS: (ESI, m / z): 490.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.08(s,1H),8.03(dd,J=10.82,1.69Hz,1H),7.35(d,J=1.50Hz,1H),7.29(t,J=8.94Hz,1H),6.99 -7.07(m,1H),6.70-6.78(m,1H),5.42(s,2H),5.13-5.24(m,2H),2.21(s,3H),1.79-1.97(m,2H),0.89(t,J=7.25Hz,3H).

[0307] Example 25. (19S)-8-(3-amino-5-fluorophenyl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 [21),2,4,6,8,10,15(20)-heptaene-14,18-dione (P27)]

[0308] Under a nitrogen atmosphere, [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (15.93 mg, 21.77 μmol), potassium carbonate (30.09 mg, 217.74 μmol), and compound P-27-1 (51.62 mg, 217.74 μmol) were added to a mixed solution of compound 8 (50 mg, 108.87 μmol) in dioxane (0.8 mL) and water (0.2 mL). After the reaction apparatus was purged three times, the reaction was carried out at 80 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Welch Ultimate C18 120×30mm×5um; mobile phase [water (0.4% trifluoroacetic acid)-methanol]; mobile phase: 45%-80% B, retention 8 min) to obtain a yellow solid compound P27 (9.3 mg, yield 16.17%). LCMS: (ESI, m / z): 490.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.11 (s, 1H) 7.95 (d, J = 10.88Hz, 1H) 7.34 (s, 1H) 6.50 (br d,J=11.76Hz,1H)6.22-6.33(m,2H)5.41(s,2H)5.18(s,2H)2.21(d,J=1.38Hz,3H)1.86(dquin,J=14.10,7.04,7.04,7.04,7.04Hz,2H)0.88(br t,J=7.25Hz,3H).

[0309] Example 26. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-(4-hydroxycyclohextrien-1-en-1-yl)-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P9)

[0310] Under a nitrogen atmosphere, 1,1-bis(diphenylphosphino)ferrocene palladium(II) dichloride (7.2 mg, 0.11 mmol), potassium carbonate (75.23 mg, 0.54 mmol), and 4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)cyclohextrien-3-en-1-ol (26.84 mg, 0.12 mmol) were added to a mixed solution of compound 8 (50.0 mg, 0.11 mmol) in dioxane (2.0 mL) and water (0.5 mL). After the reaction apparatus was purged three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30mm×10um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 25%-55% B, retention 8 min) to obtain a yellow solid compound P9 (16 mg, yield 28.3%). LCMS: (ESI, m / z): 477.1 [M+H] + . 1 H NMR(400MHz, methanol-d4)δ8.60(d,J=9.01Hz,1H),7.60-7.70(m,2H),5.65(br d,J=13.13Hz,1H),5.58(d,J=16.38Hz,1H),5.38(d,J=16.26Hz,1H),5.29(s,2H),4.11-4. 31(m,1H),2.57-2.75(m,1H),2.20-2.50(m,6H),1.85-2.18(m,4H),1.01(t,J=7.32Hz,3H).

[0311] Example 27. (19S)-8-(4-aminocyclohextrien-1-en-1-yl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P10)

[0312] Step 1: tert-Butyl N-{4-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11.0 4 , 9 .0 15 , 20 Synthesis of 2-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]cyclohextrien-3-en-1-yl}carbamate(2)

[0313] Under a nitrogen atmosphere, 1,1-bis(diphenylphosphino)ferrocene palladium(II) dichloride (7.97 mg, 0.0108 mmol), potassium carbonate (75.2 mg, 0.54 mmol), and tert-butyl N-[4-(4,4,5,5-tetramethyl-1,3,2-dioxoboropentane-2-yl)cyclohextrien-3-en-1-yl]carbamate (70.3 mg, 0.21 mmol) were added to a mixed solution of compound 8 (50 mg, 0.108 mmol) in dioxane (1.6 mL) and water (0.4 mL). After purging the reaction apparatus three times, the reaction was carried out at 100 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product 2 (65 mg). The crude product was used directly in the next step without purification. LCMS: (ESI, m / z): 576.3 [M+H] + .

[0314] Step 2: (19S)-8-(4-aminocyclohextrien-1-en-1-yl)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P10)

[0315] Trifluoroacetic acid (6 mL) was added to a solution of compound 2 (65 mg) in dichloromethane (2.0 mL) at room temperature. The reaction was carried out at 25 °C for 5 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 75×30 mm×3 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 5%-45% B, retention 8 min) to obtain a yellow solid compound P10 (16.5 mg, yield 24.7%). LCMS: (ESI, m / z): 476.1 [M+H] + . 1H NMR (400MHz, methanol-d4) δ8.51-8.70(m,1H),7.76(d,J=10.76Hz,1H),7.69(s,1H),5.76(br d,J=5.38Hz,1H),5.62(d,J=16.38Hz,1H),5.42(br d,J=16.38Hz,1H),5.33(br d,J=4.25Hz,2H),3.65-3.84(m,1H),2.75-2.89(m,1H),2.26-2.68(m,7H),1.93-2.21(m,3H),1.03(t,J=7.38Hz,3H).

[0316] Example 28. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-(3-hydroxyprop-1-yn-1-yl)-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P28)

[0317] In a glove box, methanesulfonic acid [(tricyclohexylphosphine)-2-(2-aminobiphenyl)]palladium(II) (24 mg, 32.4 μmol), N,N-dicyclohexylmethylamine (191 mg, 0.972 mmol), cuprous iodide (6 mg, 0.0324 μmol), and tert-butyldimethyl(prop-2-yn-1-oxy)silane (207.4 mg, 1.62 mmol) were added to a 6 mL solution of compound 8 (150 mg, 0.324 mmol). The reaction was carried out at 80 °C for 12 hours under an argon atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: 3-Phenomenex Luna C18 75×30mm×3um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 60%-90% B, retention 8 min) to obtain a yellow solid compound P28 (14.3 mg, yield 10%). LCMS: (ESI, m / z): 435.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.92 (s, 1H), 7.97 (d, J = 10.76Hz, 1H), 7.34 (s, 1H), 6.54 (br s,1H),5.43(s,2H),5.32(s,2H),4.56(s,2H),2.59(d,J=1.6Hz,3H),1.81-1.93(m,2H),0.88(t,J=7.25Hz,3H).

[0318] Example 29. (19S)-19-ethyl-6-fluoro-19-hydroxy-8-(4-hydroxybut-1-yn-1-yl)-7-methyl-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-14,18-dione (P29)

[0319] In a glove box, methanesulfonic acid [(tricyclohexylphosphine)-2-(2-aminobiphenyl)]palladium(II) (24.00 mg, 32.4 μmol), N,N-dicyclohexylmethylamine (191 mg, 0.972 mmol), cuprous iodide (6 mg, 0.0324 μmol), and 4-tert-butyldimethylsiloxybutyne (278 mg, 1.62 mmol) were added to a 6 mL solution of compound 8 (150 mg, 0.324 mmol). The reaction was carried out at 80 °C for 12 hours under an argon atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: 3-Phenomenex Luna C18 75×30mm×3um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 60%-90% B, retention 8 min) to obtain a yellow solid compound P29 (10.01 mg, yield 7.2%). LCMS: (ESI, m / z): 449.2 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.93(s,1H),7.93(d,J=10.76Hz,1H),7.33(s,1H),6.53(s,1H),5.43(s,2H),5.31(s,2H),5.07(t,J=5.50 Hz,1H),3.69-3.82(m,2H),2.83(t,J=6.63Hz,2H),2.58(d,J=2.0Hz,3H),1.87(tt,J=14.65,7.11Hz,2H),0.88(t,J=7.32Hz,3H).

[0320] Linker-Payload Synthesis:

[0321] Example 30.6-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)-N-{[({[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]aminocarbonyl}-2-phenylethyl]aminocarbonyl}methyl)aminocarbonyl]methyl}hexamamide (LP1)

[0322] At room temperature, 2-hydroxypyridine-N-oxide (17.67 mg, 0.16 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (30.49 mg, 0.16 mmol), and 2-[(2S)-2-(2-{2-[6-(2,5-dioxonyl-2,5-dihydro-1H-pyrrolo-1-yl)hexanoylamino]acetamido}acetamido)-3-phenylpropionylamino]acetic acid (56.16 mg, 0.11 mmol) were sequentially added to a dimethylformamide (2 mL) solution of compound P6 (50 mg, 0.11 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30mm 10µm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 25%-55% B, retention 8 min) to obtain a yellow solid compound LP1 (48.0 mg, yield 42.77%). LCMS: (ESI, m / z): 983.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ10.04(br d,J=1.25Hz,1H),8.38(br d,J=3.38Hz,1H),8.14(br dd,J=7.94,3.31Hz,1H),7.96-8.08(m,4H),7.76(br d,J=7.88Hz,1H),7.66(br s,1H),7.57(t,J=7.82Hz,1H),7.34(s,1H),7.23(d,J=4.25Hz,4H),7.16(dq,J=8.63,4.17Hz,1H),7.03(br d,J=7.50Hz,1H),6.97(s,2H),5.40(s,2H),5.15(s,2H),4.43-4.53(m,1H),3.8 3-3.97(m,2H),3.69-3.77(m,1H),3.52-3.67(m,4H),3.32-3.34(m,2H),3.05(br dd,J=13.63,4.00Hz,1H),2.81(br dd,J=13.76,10.01Hz,1H),2.17(br d,J=1.63Hz,3H),2.06-2.11(m,2H),1.78-1.91(m,2H),1.45(dq,J=14.87,7.34Hz,4H),1.11-1.21(m,2H),0.87(br t,J=7.25Hz,3H).

[0323] Example 31. (2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]-3-phenylpropionamide (LP2)

[0324] Step 1: Synthesis of tert-butyl-2-[(2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-3-phenylpropionylamino]acetate (LP-2-2)

[0325] At room temperature, 2,5-dioxoylidene-1-yl-2-bromoacetate (289 mg, 1.22 mmol) was added to a solution of LP-2-1 (400 mg, 1.02 mmol) in dimethylformamide (4 mL). The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated to give a white solid compound LP-2-2 (400 mg, crude product). LCMS: (ESI, m / z): 513.2 [M+H] + .

[0326] Step 2: Synthesis of 2-[(2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-3-phenylpropionylamino]acetic acid (LP-2-3)

[0327] Trifluoroacetic acid (3.56 mL, 47.96 mmol) was added to a dichloromethane (4 mL) solution of compound LP-2-2 (400 mg, 0.8 mmol) at 0 °C. After purging the reaction apparatus three times, the reaction was carried out at 0 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100×30 mm×5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 1%-30% B, retention 8 min) to obtain a white solid compound LP-2-3 (120 mg, yield 33.7%). LCMS: (ESI, m / z): 457.1 [M+H] + . 1H NMR(400MHz,DMSO-d6)δ12.58(br s,1H)8.51(br t,J=5.69Hz,1H)8.36(t,J=5.63Hz,1H)8.02-8.19(m,2H)7.13-7.34(m,5H)4.53(td,J=9.19,3.88Hz,1H)3.93(s,2 H)3.70-3.83(m,5H)3.58(dd,J=16.70,5.57Hz,1H)3.04(dd,J=13.63,4.00Hz,1H)2.76(dd,J=13.76,10.01Hz,1H).

[0328] Step 3: (2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]-3-phenylpropionamide (LP2)

[0329] At room temperature, 2-hydroxypyridine-N-oxide (17.67 mg, 0.16 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrobromide (37.51 mg, 0.16 mmol), and compound LP-2-3 (48.49 mg, 0.11 mmol) were sequentially added to a dimethylformamide (2 mL) solution of compound P6 (50 mg, 0.11 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100×30 mm 10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 65%-95% B, retention 8 min) to obtain a yellow solid compound LP2 (31.0 mg, yield 30.9%). LCMS:(ESI,m / z):910.0[M+H] + . 1H NMR (400MHz, DMSO-d6) δ10.06(s,1H),8.45-8.55(m,1H),8.42(br t,J=5.63Hz,1H),8.09-8.20(m,2H),7.93-8.08(m,2H),7.75(br d,J=8.51Hz,1H),7.67(br s,1H),7.57(t,J=7.88Hz,1H),7.34(s,1H),7.20-7.29(m,4H),7.12-7.20(m,1H),7.03(d,J=7.75Hz,1H),6. 35-6.65(m,1H),5.40(s,2H),5.15(s,2H),4.41-4.60(m,1H),3.82-4.00(m,4H),3.69-3.82(m,3H),3.60(br dd,J=16.51,5.38Hz,1H),3.05(dd,J=13.82,4.32Hz,1H),2.79(dd,J=13.88,10.13Hz,1H),2.18(s,3H),1.78-1.94(m,2H),0.88(t,J=7.19Hz,3H).

[0330] Example 32. (2S)-2-[(2S)-2-(2-bromoacetamido)-3-methylbutyrylamido]-6-(diethylamino)-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]hexamamide (LP3)

[0331] Step 1: Synthesis of (2S)-2-{[(tert-butoxy)carbonyl]amino}-6-(diethylamino)hexanoic acid (LP-3-2)

[0332] Acetaldehyde (2.66 g, 60.9 mmol, 5 M tetrahydrofuran solution) and sodium cyanoborohydride (3.83 mg, 60.9 mmol) were added separately to a methanol (100 mL) solution of compound LP-3-1 (5 g, 20.3 mmol) under a nitrogen atmosphere. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated to obtain a yellow oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100×30 mm×5 μm; mobile phase [water (10 mM ammonium bicarbonate)-acetonitrile]; mobile phase: 1%-18% B, retention 15 min) to obtain a white solid compound LP-3-2 (1.8 g, yield 51.4%). LCMS: (ESI, m / z): 303.2 [M+H] + 1 H NMR(400MHz, methanol-d4)δ3.94-4.06(m,1H)3.23(q,J=7.35Hz,4H)3.07-3.17(m,2H)1.62-1.93(m,4H)1.46(s,11H)1.32(t,J=7.33Hz,6H)

[0333] Step 2: Synthesis of (2S)-2-amino-6-(diethylamino)hexanoic acid (LP-3-3)

[0334] Trifluoroacetic acid (13.82 g, 121 mmol) was added to a solution of compound 27-2 (1.8 g, 5.95 mmol) in dichloromethane (10 mL) under a nitrogen atmosphere. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was concentrated to obtain a yellow oily compound LP-3-3 (1.6 g, crude product). This compound was used directly in the next step without purification. LCMS: (ESI, m / z): 203.2 [M+H] +

[0335] Step 3: Synthesis of (2S)-2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutyrylamino]-6-(diethylamino)hexanoic acid (LP-3-4)

[0336] Under a nitrogen atmosphere, 2,5-dioxylidene-1-yl(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutyl ester (2.38 g, 7.59 mmol) and N,N-diisopropylethylamine (3.27 g, 25.29 mmol) were added to a tetrahydrofuran (5.00 mL) solution of compound LP-3-3 (1.6 g, 5.06 mmol). After the reaction apparatus was purged three times, the reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated to obtain a yellow oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 150×40mm×10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 5%-35% B, retention 8 min) to obtain a white solid compound LP-3-4 (500 mg, yield 24.6%). LCMS: (ESI, m / z): 402.1 [M+H] +1 H NMR(400MHz, methanol-d4)δ4.23(dd,J=6.50,5.75Hz,1H)3.84-3.94(m,1H)3.20(q,J=7.25Hz,4H)2.99-3.14(m ,2H)2.00-2.17(m,1H)1.82-1.93(m,1H)1.65-1.80(m,3H)1.45(s,11H)1.30(t,J=7.32Hz,6H)0.87-1.03(m,6H)

[0337] Step 4: Synthesis of methyl 2-[(2S)-2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutyrylamino]-6-(diethylamino)hexanoylamino]acetate (LP-3-8)

[0338] At room temperature, 2-hydroxypyridine-N-oxide (207 mg, 1.87 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (358 mg, 1.87 mmol), and methyl 2-aminoacetate (235 mg, 1.87 mmol) were sequentially added to a dimethylformamide (5 mL) solution of compound LP-3-4 (500 mg, 1.25 mmol). The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 150 × 40 mm 7 μm; mobile phase [water (10 mM ammonium bicarbonate)-acetonitrile]; mobile phase: 10%–50% B, retention 8 min) to obtain a white solid compound LP-3-8 (350 mg, yield 59.4%).

[0339] LCMS:(ESI,m / z):473.3[M+H] +

[0340] Step 5: Synthesis of 2-[(2S)-2-[(2S)-2-{[(tert-butoxy)carbonyl]amino}-3-methylbutyrylamino]-6-(diethylamino)hexanoylamino]acetic acid (LP-3-5)

[0341] Potassium trimethylsilyl oxychloride (142 mg, 1.11 mmol) was added to a solution of compound LP-3-8 (350 mg, 0.74 mmol) in dimethyl sulfoxide (4 mL). The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was concentrated to obtain a brown oily crude product. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 150×40 mm×10 μm; mobile phase [water (10 mM ammonium bicarbonate)-acetonitrile]; mobile phase: 5%-35% B, retention 8 min) to obtain a white solid compound LP-3-5 (200 mg, yield 58.9%). LCMS: (ESI, m / z): 459.2 [M+H] +1 H NMR(400MHz, methanol-d4)δ4.45(t,J=6.87Hz,1H)3.78-3.92(m,2H)3.59-3.70(m,1H)3.31(s,2H)3.19(q,J=7.30Hz,4H)3.01-3.13(m,2H)1.98 -2.14(m,1H)1.91(dt,J=13.92,6.90Hz,1H)1.67-1.82(m,3H)1.47-1.55 (m,2H)1.45(s,9H)1.30(t,J=7.30Hz,6H)0.95(dd,J=14.97,6.80Hz,6H)

[0342] Step 6: tert-Butyl N-[(1S)-1-{[(1S)-5-(diethylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]aminocarbonyl}pentyl]aminocarbonyl}-2-methylpropyl]aminomethyl ester (LP-3-6)

[0343] At room temperature, 2-hydroxypyridine-N-oxide (21.8 mg, 0.19 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrobromide (38.6 mg, 0.19 mmol), and compound P6 (68.0 mg, 0.14 mmol) were sequentially added to a dimethylformamide (1 mL) solution of compound LP-3-5 (60 mg, 0.13 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 75 × 30 mm 3 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 65%–95% B, retention 8 min) to obtain a yellow solid compound LP-3-6 (70 mg, yield 52.1%). LCMS: (ESI, m / z): 912.5 [M+H] +1 H NMR(400MHz,DMSO-d6)δ10.05(br d,J=19.01Hz,1H)8.95(br s,1H)8.31-8.57(m,1H)7.89-8.09(m,3H)7.71-7.88(m,1H)7.63-7.70(m,1H)7.57(t,J=7.88Hz,1H)7.35(s,1H)7.04(br d,J=7.13Hz,1H)6.69(br t,J=8.69Hz,1H)6.53(br d,J=4.25Hz,1H)5.40(s,2H)5.16(br s,2H)4.17-4.32(m,1H)3.91(br d,J=5.25Hz,2H)3.76(br d,J=8.13Hz,1H)3.07(br dd,J=6.75,4.63Hz,4H)2.95(br d,J=4.63Hz,2H)2.17(br s,3H)1.76-1.95(m,3H)1.64-1.75(m,1H)1.50-1.63(m,3H)1.34(br d,J=10.01Hz,11H)1.12(br t,J=7.07Hz,6H)0.88(br t,J=7.25Hz,3H)0.66-0.84(m,6H)

[0344] Step 7: (2S)-2-[(2S)-2-amino-3-methylbutyrylamino]-6-(diethylamino)-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]hexamamide (LP-3-7)

[0345] Trifluoroacetic acid (2.15 g, 18.8 mmol) was added to a solution of compound LP-3-6 (0.07 g, 0.08 mmol) in dichloromethane (0.5 mL) under a nitrogen atmosphere. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was lyophilized to give a yellow solid compound LP-3-7 (0.6 g, crude product). LCMS: (ESI, m / z): 812.4 [M+H] +

[0346] Step 8: (2S)-2-[(2S)-2-(2-bromoacetamido)-3-methylbutyrylamido]-6-(diethylamino)-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]hexamamide (LP3)

[0347] Potassium carbonate (85.1 mg, 0.62 mmol) was added to a mixture of dichloromethane (0.5 mL) and water (3 mL) containing compound LP-3-7 (50 mg, 0.062 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 min. Acetyl bromide (37.3 mg, 0.185 mmol) was added at 0 °C, and the reaction mixture was reacted at 0 °C for 2 h. The reaction mixture was concentrated and separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP C18 100×30 mm 10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 20%-55% B, retention 8 min) to obtain a yellow solid compound LP3 (20.02 mg, yield 31.4%). LCMS: (ESI, m / z): 932.3 [M+H] + 1 H NMR (400MHz, DMSO-d6) δ10.11(s,1H),8.90(br s,1H),8.23-8.37(m,2H),8.20(br dd,J=6.94,2.69Hz,1H),7.95-8.06(m,2H),7.75(br d,J=8.13Hz,1H),7.64(s,1H),7.57(t,J=7.88Hz,1H),7.35(s,1H),7.04(br d,J=7.38Hz,1H),6.40-6.63(m,1H),5.41(s,2H),5.16(br s,2H),4.21-4.34(m,1H),4.09-4.20(m,1H),3.81-4.00(m,4H),3.01-3.11(m,4H),2.91-3.00(m,2H),2.17(br s,3H),1.78-1.97(m,3H),1.65-1.77(m,1H),1.48-1.63(m,3H),1.26-1.41(m,2H),1.13(br t,J=7.13Hz,6H),0.72-0.92(m,9H) 1H NMR (400MHz, DMSO-d6+D2O) δ10.01-10.18(m,1H),8.23-8.36(m,1H),8.20(br dd,J=7.07,5.19Hz,1H),7.95-8.03(m,2H),7.74(br d,J=8.13Hz,1H),7.61(s,1H),7.56(t,J=7.88Hz,1H),7.36(s,1H),7.04(d,J=7.50Hz,1H),5.39(s,2H),5.14(br s,2H),4.17-4.30(m,1H),4.05-4.16(m,1H),3.76-4.00(m,4H),3.00-3.12(m,4H),2.89-2.99(m,2H),2.16(t,J=2.50H z,3H),1.78-1.97(m,3H),1.64-1.76(m,1H),1.48-1.63(m,3H),1.25-1.40(m,2H),1.05-1.18(m,6H),0.70-0.91(m,9H)

[0348] Example 33. Synthesis of (2S)-2-[2-[2-(2-bromoacetamido)acetamido]acetamido]-N-[2-{[(4-{(S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-10-yl}cyclohex-3-en-1-yl)oxy]methyl}amino]-2-oxoethyl]-3-phenylpropionamide (LP4)

[0349] Step 1: Synthesis of N-tert-butoxycarbonyl-glycyl-L-phenylalanylglycylglycine (a-2)

[0350] Compound a-1 (1 g, 2.29 mmol) was dissolved in N,N-dimethylformamide (10 mL), and N,N-dicyclohexylcarboimide (567.28 mg, 2.75 mmol) and N-hydroxysuccinimide (290.05 mg, 2.52 mmol) were added to the solution. After stirring at room temperature for 3 hours, glycine (1.72 g, 22.91 mmol) was added to the system. Stirring was continued for 12 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with acetonitrile, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 250*50mm*15um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 10 min) to obtain a white solid compound a-2 (900 mg, yield 79.6%). 1 H NMR (400MHz, DMSO-d6) δ8.34 (br t, J=5.81Hz, 1H) 8.15 (br d, J=8.04Hz, 1H) 8.07 (t, J=5.69Hz, 1H) 7.90 (br t, J=5.26Hz, 1H) 7.13-7.31 (m, 5H) 7.00 (br t,J=5.75Hz,1H)4.52(td,J=8.85,4.33Hz,1H)3.66-3.79(m,6H)3.51-3.63(m ,3H)3.05(dd,J=13.79,4.27Hz,1H)2.77(dd,J=13.79,9.84Hz,1H)1.37(s,9H)

[0351] Step 2: Synthesis of (S)-7-benzyl-17,17-dimethyl-3,6,9,12,15-pentoxo-16-oxo-2,5,8,11,14-pentazaoctadecyl acetate (a)

[0352] Compound a-2 (0.5 g, 1.01 mmol) was dissolved in tetrahydrofuran (10 mL), and lead acetate (598.94 mg, 1.22 mmol) and glacial acetic acid (121.68 mg, 2.03 mmol) were added sequentially to the solution. The reaction was stirred at 50 °C for 12 hours. After the reaction was complete, water (20 mL) was added for dilution, and the mixture was extracted three times with ethyl acetate (20 mL). The organic phase was washed three times with saturated brine (10 mL). The washed organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was slurried with methyl tert-butyl ether (5 mL) to give a white solid compound a (0.4 g, yield 77.79%). 1H NMR (400MHz, DMSO-d6) δ8.83 (br t, J=6.88Hz, 1H) 8.32 (br t, J=5.69Hz, 1H) 8.14 (br d, J=8.13Hz, 1H) 7.89 (br t, J=5.00Hz, 1H) 7.11-7.35 (m, 6H) 6.98 (br t,J=5.75Hz,1H)5.05-5.16(m,2H)4.52(td,J=8.76,4.50Hz,1H)3.69-3.77(m,3H)3.52-3.58(m,2H)3.05(dd,J=13.70,4.32Hz,1H)2.77(br dd,J=13.76,9.76Hz,1H)2.00(s,3H)1.38(s,9H)

[0353] Step 3: Synthesis of ((7S)-7-benzyl-1-((4-((S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-10-yl)cyclohexyl-3-en-1-yl)oxy)-3,6,9,12-tetraoxo-2,5,8,11-tetraazatridecane-13-yl)tert-butyl carbamate (LP-4-2)

[0354] Anhydrous p-toluenesulfonic acid (20.24 mg, 117.52 μmol) was added to a solution of compound P9 (140 mg, 293.81 μmol) and compound a (298.24 mg, 587.62 μmol) in N,N-dimethylformamide (1 mL). The reaction was stirred at 40°C for 24 hours. The reaction solution was directly separated by preparative high-performance liquid chromatography (HPLC) (column type: 3-Phenomenex Luna C18 75*30mm*3um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 30%-60% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-4-2 (30 mg, yield 11%). Yellow solid compound P9 (100 mg) was also recovered. LCMS: (ESI, m / z): 924.3 [M+H] + .

[0355] Step 4: Synthesis of (2S)-2-[2-(2-aminoacetamido)acetamido]-N-[2-{[(4-{(S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-10-yl}cyclohex-3-en-1-yl)oxy]methyl}amino]-2-oxoethyl]-3-phenylpropionamide (LP-4-3)

[0356] Compound LP-4-2 (30 mg, 32.47 μmol, 1 eq) was dissolved in dichloromethane (1.5 mL), and trifluoroacetic acid (0.15 mL) was added dropwise at 0°C. The reaction was stirred at room temperature for 1 hour. After the reaction was complete, the pH of the reaction solution was adjusted to 8 with saturated sodium bicarbonate aqueous solution, resulting in the precipitation of a viscous solid. The reaction solution was decanted, and the solid was lyophilized to obtain a yellow solid compound LP-4-3 (25 mg, yield 82.1%, trifluoroacetate), which was used directly in the next step. LCMS: (ESI, m / z): 824.3 [M+H] + .

[0357] Step 5: Synthesis of (2S)-2-[2-[2-(2-bromoacetamido)acetamido]acetamido]-N-[2-{[(4-{(S)-4-ethyl-8-fluoro-4-hydroxy-9-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3',4':6,7]indolo[1,2-b]quinoline-10-yl}cyclohex-3-en-1-yl)oxy]methyl}amino]-2-oxoethyl]-3-phenylpropionamide (LP4)

[0358] Compound LP-4-3 (25 mg, 26.66 μmol) was dissolved in a mixed solution of dichloromethane (0.5 mL) and N,N-dimethylformamide (0.5 mL). N,N-diisopropylethylamine (13.78 mg, 106.62 μmol) was added to the system, followed by dropwise addition of bromoacetyl bromide (8.07 mg, 39.98 μmol) at 0°C. After the addition was complete, the reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the dichloromethane was purged with a nitrogen stream, and the remaining reaction mixture was subjected to preparative high-performance liquid chromatography (HPLC) (column type: 3-Phenomenex Luna C18 75*30mm*3um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 20%-55% acetonitrile, retention 8 minutes) to obtain crude compound LP4 (25 mg, 75% purity). The crude product was subjected to two separate chromatography steps by high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100*30*10um; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 30%-60% acetonitrile, retention 8 min) to obtain a yellow solid compound LP4 (6.8 mg, yield 27%, trifluoroacetate). LCMS: (ESI, m / z): 850.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.63 (br t, J=6.32Hz, 1H) 8.46-8.58 (m, 2H) 8.36 (br d,J=5.75Hz,1H)8.09-8.21(m,2H)7.79-7.87(m,1H)7.32(s,1H)7.16-7.26(m,4H)7.14(br d,J=4.13Hz,1H)5.58(br s,1H)5.41(br d,J=9.01Hz,2H)5.28(s,1H)5.17-5.23(m,1H)4.68-4.80(m,2H)4.46-4.55(m,1H)3.99(br d,J=2.88Hz,2H)3.92(s,3H)3.73-3.77(m,4H)3.55-3.67(m,2H)2.98-3.09(m,1H)2.69-2.85(m,2H)2.60(br s,1H)2.32-2.37(m,3H)2.24(br d,J=7.38Hz,2H)1.96-2.09(m,1H)1.80-1.91(m,2H)0.88(t,J=7.32Hz,3H).

[0359] Example 34. (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP5)

[0360] Step 1: Synthesis of tert-butyl(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyl ester (LP5-2)

[0361] Compound LP-5-1 (25 g, 58.76 mmol) was dissolved in N,N-dimethylformamide (250 mL). Under a nitrogen atmosphere, (2R,3R,4R,5S)-6-aminohexane-1,2,3,4,5-pentaol (15.97 g, 88.14 mmol), 2-hydroxypyridine-1-cation-1-ol anion (9.69 g, 146.90 mmol), and (3-{[(ethylaminoidene)methylidene]amino}propyl)dimethylamine hydrochloride (28.36 g, 146.9 mmol) were added sequentially. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was concentrated to obtain a yellow, oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 250*100mm#10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 30%-60% acetonitrile, retention 18 min) to give a white solid compound LP-5-2 (6 g, yield 55.51%). LCMS: (ESI, m / z): 589.2 [M+H] + .

[0362] Step 2: Synthesis of (2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyric acid (LP-5-3)

[0363] Trimethylsilyl trifluoromethanesulfonate (7.55 g, 33.98 mmol) was added to a solution of compound LP-5-2 (2 g, 3.40 mmol) in dichloromethane (20 mL) under a nitrogen atmosphere at 0 °C. After the reaction apparatus was purged three times, the reaction was carried out at 0 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was quenched with sodium bicarbonate (5.71 g), and the quenched reaction solution was concentrated under vacuum to obtain a yellow oily crude product. The yellow oily crude product was dissolved in water and then freeze-dried to obtain a white solid. A white solid was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 250*100mm#10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 1%-40% acetonitrile, retention 20 min) to obtain a white solid compound LP-5-3 (1 g, yield 55.27%). LCMS: (ESI, m / z): 533.3 [M+H) + . 1 H NMR(400MHz, DMSO-d6)δ7.88(d,J=7.50Hz,2H)7.83(br t,J=5.25Hz,1H)7.70(dd,J=7.25,2.75Hz,2H)7.37-7.44(m,2H)7.30-7.36(m,2H)6.67(br d,J=7.00Hz,1H)4.19-4.27(m,3H)3.69(q,J=6.34Hz,1H)3.54-3.65(m,3H)3.45-3.51(m,1H )3.36-3.44(m,2H)3.20-3.31(m,1H)2.96-3.07(m,1H)2.04-2.17(m,2H)1.72-1.97(m,2H).

[0364] Step 3: Synthesis of 1-tert-butyl-5-methyl(2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]pentanediol (LP-5-4)

[0365] Compound LP-5-3 (1 g, 1.88 mmol) and 1-tert-butyl-5-methyl(2S)-2-aminopentanediol (448.77 mg, 2.07 mmol) were dissolved in N,N-dimethylformamide (10 mL), followed by the addition of 2-hydroxypyridine-1-cationic-1-ol anion (309.82 mg, 4.69 mmol) and (3-{[(ethylaminoidene)methylidene]amino}propyl)dimethylamine hydrochloride (899.94 mg, 4.69 mmol). After purging the apparatus three times, the reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 250*70*10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 1%-70% acetonitrile, retention 24 min) to obtain a white solid compound LP-5-4 (0.8 g, yield 58.22%). LCMS: (ESI, m / z): 732.4 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.21(br d,J=7.50Hz,1H)7.89(d,J=7.38Hz,2H)7.66-7.78(m,3H)7.52(br d,J=8.13Hz,1H)7.39-7.46(m,2H)7.29-7.38(m,2H)4.74(d,J=4.50Hz,1H)4.46(d, J=5.50Hz,1H)4.37(d,J=6.00Hz,1H)4.32(t,J=5.57Hz,1H)4.19-4.28(m,4H)4.08- 4.16(m,1H)3.95-4.04(m,1H)3.34-3.65(m,9H)3.24-3.29(m,1H)2.97-3.08(m,1H) 2.33-2.39(m,2H)2.07-2.27(m,2H)1.86-2.03(m,2H)1.67-1.86(m,2H)1.38(s,9H).

[0366] Step 4: Synthesis of (2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-5-methoxy-5-oxonylvaleric acid (LP-5-5)

[0367] Compound LP-5-4 (10 g, 13.67 mmol) was dissolved in 100 mL of dichloromethane. Under a nitrogen atmosphere, the reaction apparatus was purged three times, then cooled to 0 °C using an ice-water bath. Trimethylsilyl trifluoromethanesulfonate (30.37 g, 136.65 mmol) was added. The reaction was continued at 0 °C for 1 hour under a nitrogen atmosphere. After the reaction, the solution was concentrated to obtain a brown, oily crude product. The crude product was dissolved in 100 mL of sodium bicarbonate aqueous solution. The crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 250*100 mm #10 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 0%-80% acetonitrile, retention 20 min) to obtain a white solid compound LP-5-5 (4 g, yield 42.8%). LCMS: (ESI, m / z): 676.3 [M+H] + .

[0368] Step 5: (9H-fluorene-9-yl)methyl N-[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminomethyl ester (LP-5-6)

[0369] Compound P11 (100 mg, 205.55 μmol), VA-OAc (118.78 mg, 246.66 μmol), and zinc acetate (37.71 mg, 205.55 μmol) were dissolved in toluene (1 mL). After purging the reaction apparatus three times, the reaction proceeded at 115 °C for 4 hours under a nitrogen atmosphere. After the reaction was complete, the reaction mixture was filtered, and the filter cake was washed with tetrahydrofuran (10 mL). The filtrate was concentrated to obtain a brown solid LP-5-6 (0.1 g, crude product), which was used directly in the next reaction without further purification. LCMS: (ESI, m / z): 908.2 [M+H] + .

[0370] Step 6: (2S)-2-amino-N-[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]-3-methylbutyramide (LP-5-7)

[0371] The crude compound LP-5-6 (100 mg, 110.13 μmol) and piperidine (187.55 mg, 2.20 mmol) were dissolved in N,N-dimethylformamide (2 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 4 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-45% acetonitrile, retention 8 min) to obtain the brown solid compound LP-5-7 (30 mg, yield 39.72%). LCMS: (ESI, m / z): 686.2 [M+H] + .

[0372] Step 7: Methyl(4S)-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]butyl ester (LP-5-8)

[0373] Compound LP-5-7 (60 mg, 87.50 μmol), compound LP-5-5 (59.12 mg, 87.50 μmol), and 2-hydroxypyridine-1-cation-1-ol anion (24.30 mg, 218.74 μmol) were dissolved in N,N-dimethylformamide (0.6 mL), followed by the addition of ({[3-(dimethylamino)propyl]aminoidene}methylidene)(ethyl)amine (51.66 mg, 218.74 μmol, HBr salt). The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by high performance liquid preparative chromatography (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 30%-60% B, retention 8 minutes) to obtain a white solid compound LP-5-8 (35 mg, yield 29.78%). 1 H NMR(400MHz, DMSO-d6)δ8.70(br t,J=6.25Hz,1H)8.10(br d,J=6.75Hz,1H)7.95-8.06(m,3H)7.88(d,J=7.38Hz,2H)7.75-7.81(m,1H)7.71(br t,J=6.25Hz,2H)7.54-7.64(m,2H)7.50(br d,J=7.63Hz,1H)7.37-7.44(m,2H)7.29-7.36(m,3H)7.26(br d,J=6.38Hz,2H)6.52(s,1H)5.40(s,2H)5.15(s,2H)4.75(d,J=4.50Hz,1H)4.6 2-4.70(m,2H)4.55(s,2H)4.45(d,J=5.50Hz,1H)4.37(d,J=6.00Hz,1H)4.32(br t,J=5.50Hz,2H)4.17-4.28(m,5H)4.10-4.16(m,1H)3.92-4.01(m,1H) 3.50-3.64(m,6H)3.44-3.49(m,1H)3.34-3.43(m,4H)3.22-3.29(m,1H )2.97-3.06(m,1H)2.06-2.31(m,6H)1.82-1.97(m,5H)1.67-1.79(m,2 H)1.15-1.27(m,3H)0.85-0.91(m,3H)0.76(td,J=10.91,6.57Hz,6H).

[0374] Step 8: (4S)-4-[(2S)-2-amino-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP-5-9)

[0375] Under a nitrogen atmosphere at 0°C, lithium hydroxide monohydrate (1M, 260.53 μL) was added to a mixed solvent of compound LP-5-8 (35 mg, 26.05 μmol) in tetrahydrofuran (0.56 mL) and water (0.26 mL). The reaction was carried out at 0°C under a nitrogen atmosphere for 1 hour. After the reaction was completed, the mixture was diluted with water (5.0 mL), extracted three times with ethyl acetate (10.0 mL), and the pH of the aqueous phase was adjusted to 4 with hydrochloric acid (5%). The aqueous phase was then lyophilized. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 20%-50% acetonitrile, retention 8 min) to obtain a white solid compound LP-5-9 (10 mg, yield 34.67%). LCMS: (ESI, m / z): 1107.5 [M+H] + .

[0376] Step 9: (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}methoxy)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP5)

[0377] Compound LP-5-8 (10 mg, 9.03 μmol) and 2,5-dioxonylpyrrolidone-1-yl 2-bromoacetate (10.66 mg, 45.16 μmol) were dissolved in N,N-dimethylformamide (0.2 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP5 (6 mg, yield 52.47%). LCMS: (ESI, m / z): 1229.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.13(s,1H)8.71(br t,J=6.07Hz,1H)8.48(d,J=7.88Hz,1H)8.09-8.20(m,2H)7.96-8.05(m,2H)7.74(br d,J=5.63Hz,2H)7.56-7.63(m,1H)7.50(d,J=7.88Hz,1H)7.35(s,1H)7.26(br d,J=5.00Hz,2H)6.53(s,1H)5.41(s,2H)5.16(br s,2H)4.76(d,J=4.38Hz,1H)4.60-4.71(m,2H)4.51-4.59(m,2H)4.46(d,J=5.50Hz,1H)4.35-4.40(m,1 H)4.30-4.35(m,1H)4.19-4.30(m,4H)4.08-4.18(m,1H)3.84-3.95(m,2H)3.52-3.64(m,3H)3.43-3.50 (m,1H)3.35-3.42(m,2H)3.26(dt,J=12.88,5.25Hz,1H)2.93-3.08(m,1H)2.05-2.29(m,7H)1.79-1.95 (m,5H)1.65-1.77(m,2H)1.21(d,J=7.00Hz,3H)0.88(t,J=7.25Hz,3H)0.76(td,J=10.76,6.63Hz,6H).1 H NMR(400MHz, DMSO-d6+D2O)δ8.70(br t,J=5.82Hz,1H)8.50(d,J=7.50Hz,1H)8.15(br dd,J=18.39,7.50Hz,1H)7.94-8.05(m,2H)7.71-7.80(m,1H)7.55-7.64(m,1 H)7.46-7.51(m,1H)7.38(s,1H)7.21-7.30(m,2H)5.32-5.47(m,2H)5.15(br s,2H)4.59-4.68(m,2H)4.49-4.58(m,2H)4.15-4.30(m,3H)4.05-4.13 (m,1H)3.83-3.91(m,2H)3.52-3.62(m,3H)3.43-3.50(m,1H)3.33-3.4 1(m,2H)3.21-3.29(m,1H)2.95-3.05(m,1H)2.07-2.31(m,7H)1.86(dquin,J=13.86,7.07,7.07,7.07,7.07Hz,5H)1.65-1.78(m,2H)1.20(br d,J=6.88Hz,3H)0.87(t,J=7.32Hz,3H)0.67-0.80(m,6H).

[0378] Example 35. (4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[4-({[2-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)ethyl]aminocarbonyl}({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}methyl)(methyl)aminocarbonyl]oxo})methyl)phenyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-acetaminobutyric acid (LP6).

[0379] Step 1: Synthesis of (2S)-2-[(2S)-2-amino-3-methylbutyrylamino]-5-(aminocarbonylamino)valeric acid (LP-6-2)

[0380] Compound LP-6-1 (2 g, 5.34 mmol) was dissolved in a mixed solvent of ethyl acetate (20 mL) and hydrochloric acid / ethyl acetate (20 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 16 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was concentrated under vacuum, and the crude product was slurried with methyl tert-butyl ether (10 mL) at 25 °C for 30 min. After filtration, the solid was dried under vacuum to give a yellow solid compound LP-6-2 (1.5 g, 90% yield). 1 H NMR(400MHz, DMSO-d6)8.73(d,J=7.38Hz,1H)8.18(br d,J=3.88Hz,3H)4.18-4.25(m,1H)2.97(br t,J=6.75Hz,2H)2.05-2.15(m,1H)1.68-1.80(m,1H)1.56-1.66(m,2H)1.39-1.51(m,2H)0.96(d,J=6.88Hz,6H).

[0381] Step 2: Synthesis of (2S)-2-[(2S)-2-[(2S)-5-(tert-butoxy)-2-acetamido-5-oxoylidenevalamido]-3-methylbutyrylamido]-5-(aminocarbonylamino)valerate (LP-6-4)

[0382] Compound LP-6-3 (1.87 g, 5.47 mmol) and ethyldi(propane-2-yl)amine (1.41 g, 10.94 mmol) were added to a solution of compound LP-6-2 (1.5 g, 5.47 mmol) in N,N-dimethylformamide (30 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, water (30 mL) was added to quench the reaction, and the mixture was diluted with ethyl acetate (50 mL). The mixture was extracted three times with ethyl acetate (50 mL), and the organic phase was washed three times with saturated brine (50 mL). The washed organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow solid compound LP-6-4 (1.5 g, yield 54.69%). The crude product was used directly in the next reaction. LCMS: (ESI, m / z): 502.2 [M+H] + .

[0383] Step 3: Synthesis of tert-butyl(4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[4-(1-hydroxy-2-methoxy-2-oxoethyleneethyl)phenyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-acetaminobutyl ester (LP-6-6)

[0384] Compound LP-6-4 (1.5 g, 2.99 mmol) was dissolved in N,N-dimethylformamide (15 mL), followed by the sequential addition of compound LP-6-5 (650 mg, 3.59 mmol), O-(7-azabenzotriazole-1-YL)-N,N,N,N-tetramethylurea hexafluorophosphine salt (1.19 g, 3.14 mmol), and ethyl di(propane-2-yl)amine (773.02 mg, 5.98 mmol). After purging the reaction apparatus three times, the reaction proceeded at 25 °C for 12 hours under a nitrogen atmosphere. Following the reaction, the reaction solution was filtered, and the resulting solid was slurried with methyl tert-butyl ether (15 mL) at 20 °C for 10 minutes. The filtered solid was dried under vacuum to give a yellow solid compound LP-6-6 (1 g, yield 50.3%). LCMS: (ESI, m / z): 665.3 [M+H] + .

[0385] Step 4: tert-Butyl(4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[4-(1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}methyl)(methyl)aminocarbonyl]oxo}-2-methoxy-2-oxoethylene)phenyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-acetaminobutyl ester (LP-6-7)

[0386] Compound LP-6-6 (447 mg, 672 μmol) was dissolved in N,N-dimethylformamide (28.0 mL). Di(4-nitrophenyl)carbonate (613.69 mg, 2.017 mmol) and ethyldi(propane-2-yl)amine (521.43 mg, 4.04 mmol) were added sequentially to the above reaction system. After purging the apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. Then, compound P31 (1.11 g, 2.23 mmol), 1H-1,2,3-benzotriazol-1-ol (136.29 mg, 1.00 mmol), and ethyldi(propane-2-yl)amine (521.43 mg, 4.04 mmol) were added to the above solution. After purging the apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 150*40*10um; mobile phase [water (10mM ammonium bicarbonate)-acetonitrile]; mobile phase: 34%-64% acetonitrile, retention 8 min) to obtain a white solid compound LP-6-7 (420 mg, yield 52.5%). LCMS: (ESI, m / z): 1190.8 [M+H) + .

[0387] Step 5: 2-{4-[(2S)-2-[(2S)-2-[(2S)-5-(tert-butoxy)-2-acetamido-5-oxoylidenevalamido]-3-methylbutyrylamido]-5-(aminocarbonylamino)pentanoylamido]phenyl}-2-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxoylidene-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}methyl)(methyl)aminocarbonyl]oxo}acetic acid (LP-6-8)

[0388] Under a nitrogen atmosphere at 0°C, an aqueous solution of lithium hydroxide monohydrate (1M, 3.53mL) was added to a tetrahydrofuran (82mL) and water (13.6mL) solution of compound LP-6-7 (420mg, 352.87μmol). After the reaction apparatus was purged three times, the reaction was carried out at 0°C under a nitrogen atmosphere for 1 hour. After the reaction was complete, water (55.0 mL) was added for dilution, and the mixture was extracted three times with ethyl acetate (105.0 mL). The pH of the aqueous phase was adjusted to 4 with hydrochloric acid (5%). The aqueous phase was lyophilized to obtain a white crude solid. The crude solid was then separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 25%-60% acetonitrile, retention time 8 min) to obtain a yellow solid compound LP-6-8 (190 mg, yield 45.77%). LCMS: (ESI, m / z): 1176.4 [M+H] + .

[0389] Step 6: tert-Butyl(4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[4-({[2-(2,5-dioxylidene-2,5-dihydro-1H-pyrrolo-1-yl)ethyl]aminocarbonyl}({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxylidene-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}methyl)(methyl)aminocarbonyl]oxo})methyl)phenyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-acetaminobutyl ester (LP-6-9)

[0390] Compound LP-6-8 (190 mg, 161.53 μmol), 1-(2-aminoethyl)-2,5-dihydro-1H-pyrrole-2,5-dione (114.00 mg, 645.52 μmol, HCl), and benzotriazol-1-yl-oxytripyrrole hexafluorophosphate (448.4 mg, 861.65 μmol) were dissolved in N,N-dimethylformamide (3.8 mL). Ethyl di(propane-2-yl)amine (125.26 mg, 969.08 μmol) was added to the system. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 25%-55% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-6-9 (80 mg, yield 38.14%). LCMS: (ESI, m / z): 1298.4 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.79-10.22(m,1H)8.11-8.38(m,2H)7.94-8.08(m,3H)7.69-7.86(m,2H) 7.39-7.63(m,4H)7.30-7.37(m,2H)7.23-7.29(m,2H)6.83-6.92(m,2H)6.42-6.63(m,1H)5.97(br d,J=2.13Hz,1H)5.60-5.76(m,1H)5.42(s,4H)5.15(br d,J=4.50Hz,2H)4.47-4.71(m,2H)4.27-4.44(m,2H)4.15-4.23(m,1H)3.35(td,J=6.57,3.25Hz,4H)3.12-3.25(m,2H)2.97 -3.02(m,2H)2.84-2.89(m,1H)2.10-2.26(m,4H)1.80-2.02(m,9H)1.65-1.77(m,2H)1.30-1.62(m,12H)0.76-0.97(m,9H).

[0391] Step 7: (4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[4-({[2-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)ethyl]aminocarbonyl}({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}methyl)(methyl)aminocarbonyl]oxo})methyl)phenyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-acetaminobutyric acid (LP6).

[0392] 85% phosphoric acid (4.26 g, 36.97 mmol, 2.54 mL) was added to a solution of compound LP-6-9 (80 mg, 61.62 μmol) in acetonitrile (2.8 mL). After purging the reaction apparatus three times, the reaction was carried out at 30 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: c Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-50% acetonitrile, retention 10 min) to obtain a white solid compound LP6 (25 mg, yield 29.43%). LCMS: (ESI, m / z): 621.8 [1 / 2 M + H] + . 1H NMR(400MHz,DMSO-d6)δ11.59-12.22(m,1H)9.83-10.30(m,1H)8.11-8.34(m,2H)7.92-8.09(m,3H)7.67-7.83 (m,1H)7.17-7.66(m,9H)6.81-6.95(m,2H)6.52(s,1H)5.91-6.01(m,1H)5.64-5.73(m,1H)5.41(s,4H)5.15(br d,J=5.38Hz,2H)4.46-4.80(m,2H)4.25-4.45(m,2H)4.13-4.23(m,1H)3.08 -3.31(m,4H)2.81-3.06(m,5H)2.08-2.27(m,5H)1.79-2.04(m,7H)1.71(br dd,J=7.94,4.32Hz,2H)1.51-1.63(m,1H)1.28-1.49(m,2H)0.77-0.95(m,9H).

[0393] Example 36. (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP7)

[0394] Step 1: (9H-fluorene-9-yl)methyl N-[(1S)-1-{[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]aminocarbonyl}-2-methylpropyl]aminomethyl ester (LP-7-1)

[0395] At room temperature, 2-hydroxypyridine-N-oxide (236 mg, 2.12 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (407 mg, 2.12 mmol), and compound P6 (0.50 g, 1.06 mmol) were sequentially added to a solution of (2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-3-methylbutyrylamino]propionic acid (523 mg, 1.27 mmol) in dimethylformamide (5 mL). The reaction mixture was stirred at 25 °C for 12 hours. After the reaction was complete, the reaction mixture was diluted with ethyl acetate (50 mL) and washed three times with water (50 mL). The combined organic phases were dried and concentrated to give a yellow oily compound LP-7-1 (500 mg, crude). The crude product was used directly in the next step without purification. LCMS:(ESI,m / z):864.3[M+H] + .

[0396] Step 2: (2S)-2-amino-N-[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]-3-methylbutyramide (LP-7-2)

[0397] Morpholine (1.01 g, 11.58 mmol) was added to a dimethylformamide (5 mL) solution of compound LP-7-1 (0.5 g, 0.58 mmol) under a nitrogen atmosphere. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP C18 100×30 mm 10 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 25%-50% acetonitrile, retention for 20 min) to give a yellow solid compound LP-7-2 (0.25 g, yield 67.3%). 1H NMR(400MHz,DMSO-d6)δ10.29(s,1H),8.30(br s,1H),7.93-8.07(m,2H),7.75-7.89(m,1H),7.53-7.71(m,2H),7.25-7.50(m,1 H),7.03(d,J=7.63Hz,1H),6.25-6.71(m,1H),5.40(s,2H),5.15(s,2H),4.47(br s,1H),3.66-3.72(m,1H),3.09-3.16(m,1H),2.57(br d,J=8.00Hz,1H),2.17(d,J=1.00Hz,3H),1.79-2.01(m,3H),1.28-1.39(m,3H),0.76-0.96(m,9H).

[0398] Step 3: Methyl(4S)-4-{[(1S)-1-{[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]butyl ester (LP-7-3)

[0399] At room temperature, 2-hydroxypyridine-N-oxide (86.57 mg, 0.78 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrobromide (150 mg, 0.78 mmol), and compound LP-7-2 (0.25 g, 0.39 mmol) were sequentially added to a solution of 1-9 (316 mg, 0.47 mmol) in 2.5 mL of dimethylformamide. The reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction mixture was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP C18 100×30 mm 5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 20%-60% acetonitrile, retention 8 min) to give a yellow solid compound LP-7-3 (0.2 g, yield 39.5%). LCMS:(ESI,m / z):1299.9[M+H] + . 1 H NMR(400MHz,DMSO-d6)δ10.17(br s,1H),8.22(br d,J=6.13Hz,1H),7.93-8.12(m,3H),7.66-7.93(m,7H),7.52-7.66(m,3H),7.28-7.45(m,5H),7.02(br d,J=6.75Hz,1H),6.52(s,1H),5.40(br s,2H),5.12(br s,2H),4.76(br d,J=4.50Hz,1H),4.11-4.49(m,10H),3.97(br d,J=5.50Hz,1H),3.44-3.65(m,7H),3.40(br dd,J=12.69,5.94Hz,2H),3.28(br dd,J=7.38,5.25Hz,1H),3.02(dt,J=12.66,6.36Hz,1H),2.25-2.37(m,2H),2.07-2.23(m,5H),1.69-2.02(m,7H),1.32(br d,J=5.63Hz,3H),0.72-1.00(m,9H).

[0400] Step 4: (4S)-4-[(2S)-2-amino-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .04 , 9 .0 15 , 20 Synthesis of 1-(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP-7-4)

[0401] At 0 °C, lithium hydroxide (1 M aqueous solution, 1.54 mL) was added to a mixed solution of compound LP-7-3 (200 mg, 0.15 mmol) in tetrahydrofuran (3.2 mL) and water (1.5 mL). The reaction mixture was stirred at 0 °C for 1 hour. After the reaction was complete, the reaction mixture was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP C18 100×40 mm 5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-55% acetonitrile, retention 8 min) to give a yellow solid compound LP-7-4 (0.1 g, yield 61.1%). LCMS: (ESI, m / z): 1063.4 [M+H] + .

[0402] Step 5: (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4(9),5,7,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP7).

[0403] At 25 °C, bromoacetic acid succinimide ester (26.64 mg, 112.88 μmol) was added to a mixed solution of compound LP-7-4 (0.10 g, 94.1 μmol) and dimethylformamide (1 mL). The reaction mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the reaction mixture was filtered, and the filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP C18 100×40 mm 5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-50% acetonitrile, retention 8 min) to give a white solid compound LP7 (39.27 mg, yield 35.2%). LCMS: (ESI, m / z): 1183.1 [M+H] + .

[0404] 1 H NMR (400MHz, DMSO-d6) δ12.09(s,1H),10.12-10.20(m,1H),8.48(d,J=7.88Hz,1H),8.25(br d,J=4.38Hz,1H),8.16(br d,J=7.63Hz,1H),7.95-8.07(m,2H),7.67-7.86(m,3H),7.51-7.65(m,2H),7.34(s,1H),7.02(br d,J=7.63Hz,1H),6.53(s,1H),5.40(s,2H),5.15(br s,2H),4.76(d,J=4.50Hz,1H),4.45(d,J=5.63Hz,1H),4.21-4.42(m,6H),4.13- 4.20(m,1H),3.90(d,J=1.63Hz,2H),3.50-3.65(m,3H),3.42-3.49(m,1H),3.35 -3.42(m,2H),3.21-3.29(m,1H),2.95-3.07(m,1H),2.04-2.29(m,7H),1.79-2. 01(m,5H),1.64-1.79(m,2H),1.32(dd,J=7.00,2.25Hz,3H),0.77-0.95(m,9H). 1H NMR(400MHz,DMSO-d6+D2O)δ10.16(br s,1H),8.49(br d,J=7.38Hz,1H),8.09-8.32(m,2H),7.92-8.07(m,2H),7.69-7.86(m,2H),7.48-7.64(m,2H),7.36(s,1H),7.01(br d,J=7.50Hz,1H),5.29-5.45(m,2H),5.13(br s,2H),4.04-4.43(m,4H),3.75-3.95(m,2H),3.54(br dd,J=11.19,3.06Hz,3H),3.42-3.48(m,1H),3.31-3.41(m,2H),3.20-3.29(m,1H),2.99(br dd,J=13.38,7.63Hz,1H),2.04-2.29(m,7H),1.79-2.01(m,5H),1.62-1.78(m,2H),1.26-1.37(m,3H),0.83(dq,J=13.96,7.06Hz,9H).

[0405] Example 37 (2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-N-({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)-3-phenylpropionamide (LP8)

[0406] Step 1: Synthesis of {2-[(2S)-2-{2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)acetamido]acetamido}-3-phenylpropionylamino]acetamido}methyl acetate (LP-8-1)

[0407] To a mixture of compound 2866302-04-5 (1.5 g, 2.44 mmol), copper acetate (309.79 mg, 1.71 mmol), and lead acetate (1.30 g, 2.92 mmol), N,N-dimethylformamide (21 mL) and glacial acetic acid (336.53 mg, 5.60 mmol, 320.81 μL) were added, respectively. After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 5 hours under a nitrogen atmosphere. The reaction solution was quenched with water (50 mL) and extracted three times with dichloromethane (50 mL). The organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a yellow oily crude product. The crude product was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 (250*70mm, 15um); mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 45%-75% acetonitrile, retention 20 min) to obtain a white solid compound LP-8-1 (0.7 g, yield 41.52%). LCMS: (ESI, m / z): 570.3 [M-AcO₂] + . 1 H NMR (400MHz, DMSO-d6) δ8.83 (br t, J=6.94Hz, 1H) 8.32 (br t, J=5.75Hz, 1H) 8.14 (br d,J=8.13Hz,1H)7.96-8.04(m,1H)7.89(d,J=7.38Hz,2H)7.71(d,J=7.50Hz,2H)7.58(br t,J=5.94Hz,1H)7.38-7.45(m,2H)7.29-7.36(m,2H)7.22-7.26(m,4H)7.14-7.21(m,1H)5.02 -5.14(m,2H)4.45-4.61(m,1H)4.26-4.32(m,2H)4.21-4.25(m,1H)3.72-3.81(m,3H)3.62(br d,J=6.00Hz,3H)3.05(dd,J=13.82,4.44Hz,1H)2.78(br dd,J=13.76,9.76Hz,1H)1.99(s,3H).

[0408] Step 2: (9H-fluorene-9-yl)methyl N-{[({[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 ,20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]aminocarbonyl]methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl]methyl)aminocarbonyl]methyl]aminomethyl]aminomethyl]aminomethyl ester (LP-8-2)

[0409] 4-Methylbenzene-1-sulfonic acid (19.03 mg, 110.49 μmol) was added to a solution of compound LP-8-1 (260.90 mg, 414.35 μmol) and compound P28 (120 mg, 276.23 μmol) in N,N-dimethylformamide (0.4 mL). The reaction was carried out at 40 °C for 12 hours under a nitrogen atmosphere. The reaction solution was used directly for the next reaction. LCMS: (ESI, m / z): 1004.1 [M+H] + .

[0410] Step 3: (2S)-2-[2-(2-aminoacetamido)acetamido]-N-({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)-3-phenylpropionamide (LP-8-3)

[0411] Morpholine (120.33 mg, 1.38 mmol, 121.54 μL) was added to the above solution containing compound LP-8-2. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-60% acetonitrile, retention 8 min) to obtain a white solid compound LP-8-3 (45 mg, yield 20.84%). LCMS: (ESI, m / z): 782.4 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ8.98(s,1H)8.74(br t,J=6.69Hz,1H)8.41(br t,J=5.75Hz,1H)8.27(br d,J=8.13Hz,1H)8.18(s,1H)8.01(d,J=10.51Hz,1H)7.47(d,J=8.13Hz,1H)7.34(s,1H)7.08-7.27(m,6H)6.55(br s,1H)5.43(s,2H)5.32(s,2H)4.81(br d,J=6.75Hz,2H)4.61(s,2H)4.51-4.58(m,1H)3.75-3.86(m,3H)3.63-3.69(m,1H)3.23(s,2H)3.06(br dd,J=14.01,4.13Hz,1H)2.76-2.81(m,1H)2.61(br d,J=1.50Hz,3H)1.78-1.98(m,2H)0.88(t,J=7.25Hz,3H).

[0412] Step 4: (2S)-2-{2-[2-(2-bromoacetamido)acetamido]acetamido}-N-({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)-3-phenylpropionamide (LP8)

[0413] (2,5-dioxoylide-1-yl)2-bromoacetate (67.93 mg, 287.80 μmol) was added to N,N-dimethylformamide (0.9 mL) containing compound LP-8-3 (45 mg, 57.56 μmol). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-50% acetonitrile, retention 8 min) to obtain a white solid compound LP8 (17 mg, yield 32.7%). LCMS: (ESI, m / z): 904.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.97(s,1H)8.71(br t,J=6.57Hz,1H)8.51(br t,J=5.38Hz,1H)8.36(br t,J=5.75Hz,1H)8.11-8.19(m,2H)8.00(d,J=10.51Hz,1H)7.34(s,1H)7.19-7.27(m,4H)7.13-7. 19(m,1H)6.54(s,1H)5.43(s,2H)5.32(s,2H)4.81(d,J=6.63Hz,2H)4.61(s,2H)4.47-4.56(m,1H) 3.91(s,2H)3.70-3.85(m,5H)3.55-3.67(m,1H)3.00-3.11(m,1H)2.80(dd,J=13.57,9.94Hz,1H)2 .61(d,J=1.38Hz,3H)1.87(dquin,J=14.76,7.13,7.13,7.13,7.13Hz,2H)0.88(t,J=7.32Hz,3H). 1H NMR(400MHz, DMSO+D2O-d6)δ8.94(s,1H)8.67(br t,J=6.44Hz,1H)8.53(br t,J=5.75Hz,1H)8.33(br t,J=5.50Hz,1H)8.11-8.22(m,2H)7.90(d,J=10.63Hz,1H)7.37(s,1H)7.05-7.24(m,5H)5.33-5.47(m,2H)5.25 (s,2H)4.73-4.83(m,2H)4.56(s,2H)4.39-4.48(m,1H)3.85(s,2H)3.68-3.82(m,5H)3.54-3.65(m,1H)3.03(br dd,J=13.76,4.13Hz,1H)2.77(br dd,J=13.51,9.51Hz,1H)2.55(s,3H)1.75-1.95(m,2H)0.84(t,J=7.32Hz,3H).

[0414] Example 38 ((4S)-4-(2-bromoacetamido)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP9)

[0415] Step 1: Synthesis of (Resin LP-9-S2)

[0416] 2-(9H-fluorene-9-ylmethylamino)acetic acid (29.44 g, 99.03 mmol), resin LP-9-S1 (50.00 g, 49.52 mmol), and N,N-diisopropylethylamine (64.00 g, 495.17 mmol, 86.25 mL) were added to a solid-phase column containing 400 mL of dichloromethane and reacted at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was washed three times with 300 mL of dichloromethane and three times with a 1 / 1 dichloromethane / methanol mixture. Then, N,N-dimethylformamide (400 mL) and piperidine (24.62 g, 289.08 mmol, 28.55 mL) were added separately to solid-phase columns and reacted at 25 °C for 1 hour under a nitrogen atmosphere. The reaction solution was filtered and washed twice with N,N-dimethylformamide (300 mL) to obtain LP-9-S2, which was then used directly in the next step.

[0417] Step 2: Synthesis of (Resin LP-9-S3)

[0418] N,N-Dimethylformamide (400 mL) was added to a solid-phase column containing LP-9-S2 (52.58 g, 28.96 mmol) and (2S)-5-(aminocarbonylamino)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)valeric acid (11.51 g, 28.96 mmol), followed by the sequential addition of ethyl 1-hydroxy-1H-1,2,3-triazol-4-carboxylic acid ester (23.96 g, 57.92 mmol) and N,N-diisopropylethylamine (18.71 g, 144.79 mmol, 25.22 mL). The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. After washing three times with N,N-dimethylformamide (300 mL), N,N-dimethylformamide (400 mL) and piperidine (24.66 g, 289.65 mmol, 28.61 mL) were added to the solid-phase column. The reaction was carried out at 25 °C for 1 hour under a nitrogen stream. After the reaction was completed, the reaction solution was filtered, washed three times with N,N-dimethylformamide (300 mL), and used directly for the next step.

[0419] Step 3: Synthesis of (Resin LP-9-S4)

[0420] N,N-Dimethylformamide (400 mL) was added to a solid-phase column containing LP-9-S3 (57.13 g, 28.96 mmol), (2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-3-methylbutyric acid (9.83 g, 28.95 mmol), ethyl 1-hydroxy-1H-1,2,3-triazol-4-carboxylic acid ester (22.02 g, 57.91 mmol), and N,N-diisopropylethylamine (18.71 g, 144.76 mmol, 25.22 mL). The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. The reaction solution was filtered and washed three times with N,N-dimethylformamide (300 mL). Then, N,N-dimethylformamide (400 mL) and piperidine (24.65 g, 289.45 mmol, 28.59 mL) were added separately to the solid-phase system. The reaction was carried out at 25 °C for 1 hour under a nitrogen stream. The reaction solution was filtered and washed three times with N,N-dimethylformamide (300 mL) before being used directly in the next step.

[0421] Step 4: Synthesis of (2-[(2S)-2-[(2S)-2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]-3-methylbutyrylamido]-5-(aminocarbonylamino)valamido]acetic acid)(LP-9-1)

[0422] N,N-Dimethylformamide (400 mL) was added to a solid-phase column containing (2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxonylpentanoic acid (12.32 g, 28.94 mmol), compound LP-9-S4 (10.00 g, 28.94 mmol), N,N-diisopropyl-ethylamine (18.70 g, 144.72 mmol, 25.21 mL), and ethyl 1-hydroxy-1H-1,2,3-triazol-4-carboxylic acid ester (23.95 g, 57.89 mmol). The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. The reaction solution was filtered, washed twice with N,N-dimethylformamide (400 mL), and twice with dichloromethane (400 mL). The resin was washed five times with trifluoroacetic acid / dichloromethane (5%, 5 × 300 mL). Each fraction was poured into ice water (300 mL) and neutralized to pH 6 with a saturated sodium carbonate aqueous solution. After concentrating and removing the dichloromethane, filtration yielded a white solid compound LP-9-1 (9 g, 65% yield). LCMS: (ESI, m / z): 739.4 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.2-13.2(m,1H),8.0-8.2(m,2H),7.89(br d,2H,J=7.4Hz),7.7-7.8(m,3H),7.61(br d,1H,J=8.3Hz),7.4-7.5(m,2H),7.3-7.4(m,2H),5.98(br s,1H),5.41(br s,2H),4.1-4.4(m,5H),4.0-4.1(m,1H),3.5-3.8(m,2H),2.9-3.0(m,2H),2.21(br s,2H),1.8-2.1(m,2H),1.6-1.8(m,2H),1.3-1.5(m,12H),0.81(br dd,6H,J=6.7,12.0Hz).

[0423] Step 5: Synthesis of (tert-butyl(4S)-4-{[(1S)-1-{[(1S)-1-{[(acetoxy)methyl]aminocarbonyl}-4-(aminocarbonylamino)butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester)(LP-9-2)

[0424] 2-[(2S)-2-[(2S)-2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]-3-methylbutyrydoamino]-5-(aminocarbonylamino)valamido]acetic acid (6.5 g, 8.80 mmol), copper acetate (1.12 g, 6.16 mmol), and lead acetate (11.70 g, 26.39 mmol) were added to N,N-dimethylformamide (130 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was diluted with water (50 mL) and ethyl acetate (100 mL) and filtered. The filtrate was extracted three times with ethyl acetate (100 mL), and the organic phase was washed three times with saturated brine (100 mL). The solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a white solid crude product. This crude product was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*70 mm, 15 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 45%-75% acetonitrile, retention 20 min) to obtain a white solid compound LP-9-2 (2.3 g, yield 34.7%). LCMS: (ESI, m / z): 775.2 [M+Na] + . 1H NMR(400MHz, DMSO-d6)δ8.89(t,J=7.03Hz,1H)8.10(br d,J=7.63Hz,1H)7.90(d,J=7.63Hz,2H)7.67-7.76(m,3H)7.58(br d,J=8.46Hz,1H)7.39-7.46(m,2H)7.27-7.38(m,2H)5.93(br t,J=5.36Hz,1H)5.38(s,2H)4.99-5.17(m,2H)4.16-4.34(m,5H)4.07(td,J=8.49,5.66Hz,1H)2.94(dquin,J=12.11,6.15,6.15,6.15, 6.15Hz,2H)2.14-2.29(m,2H)1.81-2.04(m,5H)1.65-1.79(m,1H)1.44-1.64(m,2H)1.25-1.43(m,11H)0.82(dd,J=12.40,6.79Hz,6H).

[0425] Step 6: (4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyric acid)(LP-9-3)

[0426] Compound P28 (0.8 g, 1.84 mmol), compound LP-9-2 (1.66 g, 2.21 mmol), and scandium trifluoromethanesulfonate (906.34 mg, 1.84 mmol) were added to 1,2-dichloroethane (16 mL). After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was concentrated to obtain a yellow solid, compound 9 (0.8 g, crude). The yellow solid compound LP-9-3 was used directly in the next step. LCMS: (ESI, m / z): 1071.2 [M+H] + .

[0427] Step 7: ((4S)-4-amino-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP-9-4)

[0428] Compound LP-9-3 (0.8 g, 709.71 μmol) and morpholine (618.30 mg, 7.10 mmol, 624.55 μL) were added to N,N-dimethylformamide (16 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-35% acetonitrile, retention 8 min) to obtain a white solid compound LP-9-4 (370 mg, yield 18.42%). LCMS: (ESI, m / z): 849.3 [M+H] + .

[0429] Step 8: ((4S)-4-(2-bromoacetamido)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP9)

[0430] Compound LP-9-4 (0.125 g, 147.25 μmol) and 2,5-dioxoylide-1-yl-2-bromoacetate (173.77 mg, 736.27 μmol) were added to N,N-dimethylformamide (2.5 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-55% acetonitrile, retention 8 min) to obtain a white solid compound LP9 (30 mg, yield 20.1%). LCMS: (ESI, m / z): 969.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ11.92-12.24(m,1H)8.98(s,1H)8.82(br t,J=6.57Hz,1H)8.47(d,J=8.00Hz,1H)8.13(br d,J=7.13Hz,1H)8.00(d,J=10.76Hz,1H)7.87(br d,J=8.38Hz,1H)7.35(s,1H)6.53(s,1H)5.94(br t,J=5.82Hz,1H)5.32-5.45(m,5H)4.74-4.85(m,2H)4.58(s,2H)4.33-4.41(m,1H )4.14-4.27(m,2H)3.84-3.95(m,2H)2.89-3.02(m,2H)2.53-2.63(m,4H)2.23(br t,J=7.94Hz,2H)1.81-2.02(m,4H)1.61-1.78(m,2H)1.50-1.61(m,1H)1.30-1.48(m,2H)0.75-0.94(m,9H).

[0431] Example 39 ((4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP10)

[0432] Step 1: ((4S)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-(2-{2-[2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)ethoxy]ethoxy}acetamido)butyric acid)(LP-10-1)

[0433] Ethyldi(propane-2-yl)amine (44.76 mg, 346.34 μmol, 60.33 μL) was added to a solution of compound LP-9-4 (245 mg, 288.62 μmol) and compound B (167.10 mg, 346.34 μmol) in N,N-dimethylformamide (4.9 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was used directly for the next step. LCMS: (ESI, m / z): 1216.2 [M+H] +

[0434] Step 2: ((4S)-4-{2-[2-(2-aminoethoxy)ethoxy]acetamido}-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP-10-2)

[0435] Morpholine (175.49 mg, 2.01 mmol, 177.26 μL) was added to N,N-dimethylformamide (4.9 mL) containing compound LP-10-1 (245 mg, 201.44 μmol). After purging the reaction apparatus three times, the reaction was carried out at 65 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40 mm*5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-35% acetonitrile, retention 8 min) to obtain a white solid compound (80 mg, yield 11%). LCMS: (ESI, m / z): 994.4 [M+H] +

[0436] Step 3: ((4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-{[(1S)-1-{[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid)(LP10)

[0437] Compound LP-10-2 (80 mg, 80.48 μmol) and 2,5-dioxoylide-1-yl-2-bromoacetate (94.98 mg, 402.40 μmol) were added to N,N-dimethylformamide (1.6 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-55% B, retention 8 min) to obtain a white solid compound LP10 (10 mg, yield 11.14%). LCMS: (ESI, m / z): 1116.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.10(br s,1H)8.97(s,1H)8.83(br t,J=6.69Hz,1H)8.32(br t,J=4.75Hz,1H)8.16(br d,J=7.25Hz,1H)8.00(d,J=10.63Hz,1H)7.93(br d,J=8.50Hz,1H)7.74(d,J=8.13Hz,1H)7.35(s,1H)6.53(s,1H)5.94(br t,J=5.82Hz,1H)5.43(s,2H)5.39(s,2H)5.33(s,2H)4.72-4.89(m,2H)4.58(s,2H)4.3 9-4.47(m,1H)4.16-4.28(m,2H)3.92(s,2H)3.85(s,2H)3.53-3.64(m,4H)3.40-3.48( m,2H)3.24(q,J=5.21Hz,2H)2.90-3.03(m,2H)2.57-2.64(m,3H)2.18-2.26(m,2H)1.7 3-2.04(m,5H)1.61-1.71(m,1H)1.49-1.61(m,1H)1.30-1.48(m,2H)0.76-0.94(m,9H).

[0438] Example 40. (4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9.0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}butyric acid (LP11)

[0439] Step 1: Synthesis of resin LP-11-S2

[0440] 2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)acetamido]acetic acid (17.715 g, 49.99 mmol), resin LP-11-S1 (0.03 g, 49.99 mmol), and N,N-diisopropylethylamine (64.66 g, 499 mmol, 87.15 mL) were added to a solid column containing 180 mL of dichloromethane. The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the resin was washed three times with 300 mL of dichloromethane and three times with a 1 / 1 ratio of dichloromethane and methanol. The resin was dried to obtain 50 g of yellow solid resin LP-11-S2. LP-11-S2 was used directly in the next step.

[0441] Step 2: Synthesis of resin LP-11-S3

[0442] Piperidine (42.58 g, 500 mmol, 49.38 mL) was added to a solid-phase column of resin LP-11-S2 (50 g, 50 mmol) and N,N-dimethylformamide (200 mL). The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The resulting resin was washed three times with N,N-dimethylformamide (100 mL), and then N,N-dimethylformamide (200 mL), (2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-3-phenylpropionic acid (29.06 g, 75 mmol), ethyl 1-hydroxy-1H-1,2,3-triazol-4-carboxylic acid ester (41.37 g, 100 mmol), and N,N-diisopropylethylamine (32.31 g, 250 mmol, 43.54 mL) were added sequentially to the solid-phase column. The reaction was carried out at 25°C for 2 hours under a nitrogen stream. The reaction solution was filtered and washed twice with 300 mL of N,N-dimethylformamide. A yellow solid resin, LP-11-S3 (50 g), was obtained. LP-11-S3 was used directly in the next step.

[0443] Step 3: Synthesis of 2-{2-[(2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-methoxy-5-oxoylidenevalamido]-3-phenylpropionamido]acetamido}acetic acid (LP-11-1)

[0444] Piperidine (42.58 g, 500 mmol, 49.38 mL) was added to a solid column of N,N-dimethylformamide (200 mL) in resin LP-11-S3 (50 g, 50 mmol) and reacted at 25 °C for 2 hours under a nitrogen stream. After the reaction was complete, the reaction solution was filtered. The resulting resin was washed three times with N,N-dimethylformamide (100 mL), and then N,N-dimethylformamide (200 mL), (2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-methoxy-5-oxonylpentanoic acid (21.09 g, 55 mmol), ethyl 1-hydroxy-1H-1,2,3-triazol-4-carboxylic acid ester (41.37 g, 100 mmol), and N,N-diisopropylethylamine (32.31 g, 250 mmol, 43.54 mL) were added sequentially to the solid-phase column, and the reaction was carried out at 25 °C for 2 hours under a nitrogen stream. The reaction solution was filtered, washed twice with N,N-dimethylformamide (400 mL), and twice with dichloromethane (400 mL). The resin was washed five times with trifluoroacetic acid / dichloromethane (5%, 5 × 300 mL). The yellow, oily crude product obtained by concentrating the filtrate was diluted with water and freeze-dried to give a yellow solid. The solid was then slurried with methyl tert-butyl ether to obtain compound LP-11-1 (11 g, yield 34.13%). LCMS:(ESI, m / z): 645.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.32-12.98(m,1H)8.33(br t,J=5.44Hz,1H)8.00-8.10(m,2H)7.89(d,J=7.50Hz,2H)7.71(br t,J=6.88Hz,2H)7.50(br d,J=8.00Hz,1H)7.37-7.46(m,2H)7.29-7.36(m,2H)7.09-7.26(m,5H)4.51-4.65(m,1H)4.08-4.35(m,3H)3.92-4.03(m,1H)3.73(br d,J=5.75Hz,1H)3.51-3.63(m,5H)3.00-3.18(m,2H)2.82(br dd,J=13.70,9.44Hz,1H)2.22(br t,J=7.82Hz,2H)1.65-1.90(m,2H).

[0445] Step 4: Synthesis of methyl (4S)-4-{[(1S)-1-[({[(acetoxy)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester (LP-11-2)

[0446] Copper acetate (2.17 g, 11.94 mmol) and lead acetate (22.70 g, 51.19 mmol) were added to N,N-dimethylformamide (220 mL) containing compound LP-11-1 (11 g, 17.06 mmol). The reaction apparatus was purged three times, and the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was diluted with water (200 mL), extracted three times with ethyl acetate (400 mL), and washed five times with saturated brine (300 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a yellow, oily crude product. This crude product was then separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 250mm*100mm*15um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 40%-75% acetonitrile, retention 23 min) to obtain a white solid compound LP-11-2 (5 g, yield 44.49%). LCMS: (ESI, m / z): 681.0 [M+Na] + 1 H NMR(400MHz, DMSO-d6)δ8.83(br t,J=6.88Hz,1H)8.33(br t,J=5.50Hz,1H)8.04(br d,J=7.88Hz,1H)7.89(d,J=7.50Hz,2H)7.71(br t,J=6.50Hz,2H)7.49(br d,J=8.00Hz,1H)7.39-7.44(m,2H)7.29-7.35(m,2H)7.18-7.24(m,4H)7.11-7.16(m,1H)5.10(br d,J=7.13Hz,2H)4.47-4.60(m,1H)4.18-4.34(m,3H)3.94-4.02(m,1H)3.77(br d,J=5.88Hz,1H)3.74(br d,J=5.88Hz,1H)3.58(s,3H)3.05(br dd,J=13.82,4.44Hz,1H)2.82(br dd,J=13.82,9.44Hz,1H)2.22(br t,J=7.82Hz,2H)1.98(s,3H)1.77-1.87(m,1H)1.66-1.76(m,1H)

[0447] Step 5: Methyl(4S)-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester (LP-11-3)

[0448] Compound LP-11-2 (833.95 mg, 1.27 mmol), P28 (0.5 g, 1.15 mmol), and 4-methylbenzene-1-sulfonic acid (198.20 mg, 1.15 mmol) were added to N,N-dimethylformamide (10 mL). After purging the reaction apparatus three times, the reaction was carried out at 40 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 35%-80% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-11-3 (240 mg, yield 20.18%). LCMS: (ESI, m / z): 1033.0 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ8.90(s,1H)8.72(br t,J=6.50Hz,1H)8.30(br t,J=5.44Hz,1H)8.15(br d,J=7.88Hz,1H)7.97(d,J=10.63Hz,1H)7.74(br dd,J=13.26,7.50Hz,2H)7.65(br t,J=7.25Hz,2H)7.57(br d,J=7.25Hz,1H)7.16-7.38(m,9H)7.08-7.15(m,1H)6.54(s,1H)5.44(br d,J=2.00Hz,2H)5.23(s,2H)4.71-4.84(m,2H)4.47-4.64(m,3H)4.24-4.32(m ,1H)4.10-4.22(m,2H)3.83-4.00(m,2H)3.67-3.77(m,1H)3.57(s,3H)3.09(br dd,J=13.70,4.44Hz,1H)2.78-2.88(m,1H)2.58(s,3H)2.14-2.24(m,2H)1.69-1.94(m,4H)0.88(t,J=7.25Hz,3H).

[0449] Step 6: (4S)-4-amino-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}butyric acid (LP-11-4)

[0450] A 1M, 1.79 mL aqueous solution of lithium hydroxide was added dropwise to a mixture of tetrahydrofuran (3.84 mL) and water (1.8 mL) containing LP-11-3 (240 mg, 232.32 μmol) at 0 °C. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40 mm*5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP-11-4 (100 mg, yield 38.9%). LCMS: (ESI, m / z): 797.0 [M+H] + .

[0451] Step 7: (4S)-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}-4-(2-{2-[2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)ethoxy]ethoxy}acetamido)butyric acid (LP-11-5)

[0452] N,N-Diisopropylethylamine (17.52 mg, 135.54 μmol, 23.61 μL) was added to a mixture of compound LP-11-4 (90 mg, 112.95 μmol) and 2,5-dioxonylpyrrolidone-1-yl 2-{2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)ethoxy]ethoxy}acetate (65.40 mg, 135.54 μmol) in N,N-dimethylformamide (1.8 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. The reaction solution was filtered, and the filtrate was used directly in the next step. LCMS: (ESI, m / z): 1164.3 [M+H] + .

[0453] Step 8: (4S)-4-{2-[2-(2-aminoethoxy)ethoxy]acetamido}-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}butyric acid (LP-11-6)

[0454] Morpholine (67.35 mg, 773.07 μmol, 68.03 μL) was added to N,N-dimethylformamide (1.8 mL) containing compound LP-11-5 (90 mg, 77.31 μmol). The reaction apparatus was purged three times and then reacted at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction, the reaction solution was filtered. The filtrate was subjected to preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-50% acetonitrile, retention 8 min) to obtain a white solid compound LP-11-6 (50 mg, yield 68.66%). LCMS: (ESI, m / z): 942.2 [M+H] +

[0455] Step 9: (4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-{[(1S)-1-[({[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}methyl)aminocarbonyl]-2-phenylethyl]aminocarbonyl}butyric acid (LP11)

[0456] Compound LP-11-6 (50 mg, 53.08 μmol) and 2,5-dioxoylide-1-yl-2-bromoacetate (62.64 mg, 265.41 μmol) were added to a solution of N,N-dimethylformamide (1 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100*30mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 8 min) to obtain a yellow solid compound LP11 (28.3 mg, yield 48.15%). LCMS: (ESI, m / z): 1064.3 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ11.96-12.18(m,1H)8.93-9.04(m,1H)8.69-8.77(m,1H)8.38(br t,J=5.44Hz,1H)8.28-8.34(m,1H)8.25(br d,J=7.63Hz,1H)8.00(d,J=10.51Hz,1H)7.56-7.70(m,1H)7.34(s,1H)7.22(d,J=4.13Hz,4H)7.10-7.18(m,1H)6.53(s,1H)5.39-5.47 (m,2H)5.32(s,2H)4.76-4.86(m,2H)4.61(s,2H)4.53(td,J=8.66,4.82Hz,1H)4.24-4.37(m,1H)3.88(s,2H)3.82-3.85(m,2H)3.79(br t,J=5.07Hz,2H)3.50-3.57(m,4H)3.39-3.44(m,2H)3.22(q,J=5.42Hz,2H)3.07(dd,J=13.45,4.44Hz,1H)2.81(br dd,J=13.82,10.07Hz,1H)2.61(d,J=1.75Hz,3H)2.10-2.18(m,2H)1.79-1.94(m,3H)1.67-1.77(m,1H)0.88(t,J=7.38Hz,3H).

[0457] Example 41. (2S)-2-[(2S)-2-[(2S)-2-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)propionylamino]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP12)

[0458] Step 1: Synthesis of (Resin LP-12-S2)

[0459] 2-(9H-fluorene-9-ylmethylamino)acetic acid (35.2 g, 120 mmol), resin LP-12-S1 (100 g, 100 mmol), and N,N-diisopropylethylamine (124 g, 1.00 mol) were added to a solid-phase column containing dichloromethane (500 mL) and reacted at 25 °C for 12 h under a nitrogen atmosphere. After the reaction, the reaction solution was filtered, and the resin was washed three times with dichloromethane (300 mL) and three times with a dichloromethane / methanol ratio of 1 / 1 (300 mL). Then, N,N-dimethylformamide (400 mL) and piperidine (85 g, 200 mmol) were added separately to the solid-phase column and reacted at 25 °C for 1 h under a nitrogen atmosphere. The reaction solution was filtered and washed twice with N,N-dimethylformamide (300 mL) to obtain LP-12-S2 (107 g), which was used directly in the next step.

[0460] Step 2: Synthesis of (Resin LP-12-S3)

[0461] 400 mL of N,N-dimethylformamide was added to a solid-phase column containing LP-12-S2 resin (107 g, 100 mmol) and (2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-3-[(triphenylmethyl)aminocarbonyl]propionic acid (71.6 g, 120 mmol), followed by the sequential addition of 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate (82.7 g, 200 mmol) and N,N-diisopropylethylamine (64.6 g, 1.00 mol). The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, washed three times with 300 mL of N,N-dimethylformamide, and the resulting LP-12-S3 (140 g) was used directly for the next step.

[0462] Step 3: Synthesis of (Resin LP-12-S4)

[0463] Under a nitrogen atmosphere, N,N-dimethylformamide (400 mL) and piperidine (34.5 g, 405 mmol) were added to a solid-phase column (LP-12-S3, 40.6 g, 40.6 mmol). The reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, washed three times with N,N-dimethylformamide (300 mL), and then a mixed solution of compound 63-1 (18.63 g, 48.72 mmol), 2-hydroxypyridine N-oxide (11.28 g, 101 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (19.5 g, 101 mmol) in N,N-dimethylformamide (400 mL) was added to the solid-phase column. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. The reaction solution was filtered and washed three times with N,N-dimethylformamide (300 mL). The resulting LP-12-S4 (15 g) was used directly in the next step.

[0464] Step 4: Synthesis of 2-[(2S)-3-aminocarbonyl-2-[(2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)propionylamino]propionylamino]propionylamino]acetic acid (LP-12-1)

[0465] LP-12-S4 resin (15 g) was washed five times with trifluoroacetic acid / dichloromethane (50%, 5 × 300 mL), 10 mins each time. The combined distillates were concentrated under vacuum, slurried once with methanol (50 mL), and the solid was slurried once with water (100 mL). Filtration yielded a white solid compound LP-12-1 (3.6 g, yield 34.51%). LCMS: (ESI, m / z): 554.0 [M+H] + 1H NMR (400MHz, DMSO-d6) δ11.80-13.20(m,1H),8.07(br dd,J=14.88,7.50Hz,2H),7.89(br d,J=7.50Hz,3H),7.72(br t,J=7.25Hz,2H),7.52(br d,J=7.63Hz,1H),7.38-7.46(m,2H),7.28-7.37(m,3H),6.90(br s,1H),4.55(q,J=6.96Hz,1H),4.14-4.38(m,4H),4.00-4.13(m,1H),3.70-3.81(m,2H),2.54(br d,J=5.75Hz,1H),2.40-2.48(m,1H),1.16-1.28(m,6H)

[0466] Step 5: Synthesis of [(2S)-3-aminocarbonyl-2-[(2S)-2-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)propionylamino]propionylamino]propionylamino]methyl acetate (LP-12-2)

[0467] LP-12-1 (0.05 g, 0.09 mmol), copper acetate (11.48 mg, 0.063 mmol), and lead acetate (120 mg, 0.27 mmol) were added to N,N-dimethylformamide (1.00 mL). After the reaction apparatus was purged three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. The reaction was scaled up in parallel to 66 batches. After the reaction was completed, the combined reaction solution was diluted with water (50 mL) and acetonitrile (50 mL) and filtered through diatomaceous earth. The filtrate was concentrated to obtain a yellow solid crude product, which was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*70 mm, 15 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 25%-55% acetonitrile, retention 20 min) to obtain a white solid compound LP-12-2 (0.5 g, yield 11.8%). LCMS:(ESI,m / z):590.2[M+Na] + . 1H NMR (400MHz, DMSO-d6) δ12.15-12.86(m,1H),8.07(br dd,J=14.70,7.44Hz,2H),7.84-7.97(m,3H),7.72(br t,J=7.19Hz,2H),7.52(br d,J=7.50Hz,1H),7.14-7.45(m,6H),6.90(br s,1H),4.55(q,J=7.00Hz,1H),4.01-4.34(m,5H),3.65-3.85(m,2H),2.38-2.50(m,2H),1.14-1.32(m,6H).

[0468] Step 6: (9H-fluorene-9-yl)methyl N-[(1S)-1-{[(1S)-1-{[(1S)-2-aminocarbonyl-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminomethyl ester (LP-12-3)

[0469] Compound P28 (0.38 g, 0.88 mmol), compound LP-12-2 (0.5 g, 0.88 mmol), and scandium trifluoromethanesulfonate (433 mg, 0.88 mmol) were added to 1,2-dichloroethane (10 mL). After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 3 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was concentrated to obtain a yellow solid compound LP-12-3 (0.3 g, crude). The yellow solid compound LP-12-3 was used directly in the next step. LCMS: (ESI, m / z): 942.4 [M+H] +

[0470] Step 7: (2S)-2-[(2S)-2-[(2S)-2-aminopropionylamino]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .04 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP-12-4)

[0471] Morpholine (767 mg, 8.81 mmol) was added to N,N-dimethylformamide (5 mL) containing LP-12-3 (0.30 g, crude). After purging the reaction apparatus three times, the reaction was carried out at 25°C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 250*70mm*10um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 18 min) to obtain a white solid compound LP-12-4 (225 mg, yield 12.78%). LCMS: (ESI, m / z): 720.2 [M+H] + .

[0472] Step 8: (9H-fluorene-9-yl)methyl N-{2-[2-({[(1S)-1-{[(1S)-1-{[(1S)-2-aminocarbonyl-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]} 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}methoxy)ethoxy]ethyl}aminomethyl ester (LP-12-5)

[0473] At 20 °C, 2,5-dioxylidene-1-yl 2-{2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)ethoxy]ethoxy}acetate (226.25 mg, 0.47 mmol) and N,N-diisopropylethylamine (48.5 mg, 0.38 mmol) were added to a solution of compound LP-12-4 (225 mg, 0.3 mmol) in N,N-dimethylformamide (50 mL). The reaction mixture was stirred at 20 °C for 1 hour. After the reaction was complete, the reaction mixture was used directly for the next step. LCMS: (ESI, m / z): 1087.4 [M+H] + .

[0474] Step 9: (2S)-2-[(2S)-2-[(2S)-2-{2-[2-(2-aminoethoxy)ethoxy]acetamido}propionylamino]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP-12-6)

[0475] Morpholine (544 mg, 0.06 mmol) was added to the reaction solution of compound LP-12-5. After the reaction apparatus was purged three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 8 min) to obtain a white solid compound LP-12-6 (100 mg, yield 11.4%). LCMS: (ESI, m / z): 865.2 [M+H] +

[0476] Step 10: (2S)-2-[(2S)-2-[(2S)-2-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)propionylamino]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2, 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP12)

[0477] Under a nitrogen atmosphere, bromoacetic acid succinimide ester (27.3 mg, 115 μmol) was added to N,N-dimethylformamide (1 mL) containing compound LP-12-6 (0.10 g, 115 μmol). After the reaction apparatus was purged three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP12 (30 mg, yield 26.3%).

[0478] LCMS:(ESI,m / z):985.25[M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.97 (s, 1H) 8.58 (br t, J = 6.63Hz, 1H) 8.32 (br d,J=6.88Hz,2H)8.09(d,J=7.75Hz,1H)7.99(d,J=10.63Hz,1H)7.67(d,J=7.50Hz,1H)7.39(br s,1H)7.34(s,1H)6.95(br s,1H)6.53(s,1H)5.43(s,2H)5.32(s,2H)4.78(br d,J=6.63Hz,2H)4.58(s,2H)4.49-4.55(m,1H)4.38(quin,J=7.10Hz,1H)4.27(quin,J=7.04Hz,1H)3.89(s ,2H)3.85(s,2H)3.52-3.63(m,4H)3.41-3.46(m,2H)3.24(q,J=5.71Hz,2H)2.60(d,J=1.50Hz,3H)2.55(br d,J=6.25Hz,2H)1.87(tquin,J=14.54,14.54,7.14,7.14,7.14,7.14Hz,2H)1.19-1.28(m,6H)0.88(t,J=7.25Hz,3H).

[0479] Example 42. (2S)-2-[(2S)-2-[2-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)acetamido]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP13)

[0480] Step 1: Synthesis of (Resin LP-13-S4)

[0481] Under a nitrogen atmosphere, N,N-dimethylformamide (400 mL) and piperidine (34.5 g, 405 mmol) were added to an LP-12-S3 solid-phase column (40.6 g, 40.6 mmol). The reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, washed three times with N,N-dimethylformamide (300 mL), and then compound 2 (17.9 g, 48.72 mmol), 2-hydroxypyridine N-oxide (11.28 g, 101 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (19.5 g, 101 mmol) were mixed in N,N-dimethylformamide (400 mL) and added to the solid-phase column. The reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. The reaction solution was filtered and washed three times with N,N-dimethylformamide (300 mL). The resulting LP-13-S4 (25 g) was used directly in the next step.

[0482] Step 2: Synthesis of 2-[(2S)-3-aminocarbonyl-2-[(2S)-2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)acetamido]propionamide]propionamide]acetic acid (LP-13-1)

[0483] Resin LP-13-S4 (25 g) was washed five times with trifluoroacetic acid / dichloromethane (1 / 1, 5 × 100 mL). The combined distillates were concentrated under vacuum, and the resulting solid was slurried once with methanol (50 mL) and once with water (100 mL). Filtration yielded a white solid compound LP-13-1 (2.6 g, yield 11.87%). LCMS: (ESI, m / z): 540.0 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ12.56(s,1H),8.03-8.19(m,2H),7.80-7.93(m,3H),7.71(br d,J=7.38Hz,2H),7.54(br t,J=5.88Hz,1H),7.38-7.46(m,2H),7.25-7.38(m,3H),6.88(br s,1H),4.53-4.65(m,1H),4.16-4.36(m,4H),3.60-3.84(m,4H),2.52-2.62(m,1H),2.32-2.46(m,1H),1.21(br d,J=7.00Hz,3H).

[0484] Step 3: Synthesis of [(2S)-3-aminocarbonyl-2-[(2S)-2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)acetamido]propionylamino]propionylamino]methyl acetate (LP-13-2)

[0485] LP-13-1 (0.1 g, 0.18 mmol), copper acetate (22.9 mg, 0.12 mmol), and lead acetate (120 mg, 0.54 mmol) were added to N,N-dimethylformamide (2.00 mL). After the reaction apparatus was purged three times, the reaction was carried out at 0 °C for 1 hour under a nitrogen atmosphere (the reaction was scaled up in parallel for 49 batches). After the reaction was completed, the combined reaction solution was diluted with a mixture of water (50 mL) and acetonitrile (50 mL) and filtered through diatomaceous earth. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*70 mm, 15 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 25%-60% acetonitrile, retention for 20 min) to obtain a white solid compound LP-13-2 (0.8 g, yield 13.2%). 1H NMR (400MHz, DMSO-d6) δ8.58(br t,J=6.84Hz,1H),8.02-8.20(m,2H),7.89(d,J=7.53Hz,2H),7.71(d,J=7.40Hz,2H),7.53(br t,J=5.96Hz,1H),7.39-7.45(m,2H),7.25-7.39(m,3H),6.90(br s,1H),5.06(qd,J=9.85,7.09Hz,2H),4.48-4.60(m,1H),4.18-4.35(m,4H),3.65(br d,J=5.77Hz,2H),2.53(br d,J=5.77Hz,1H),2.37-2.45(m,1H),1.96(s,3H),1.20(br d,J=7.03Hz,3H).

[0486] Step 4: (9H-fluorene-9-yl)methyl N-({[(1S)-1-{[(1S)-2-aminocarbonyl-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}methyl)aminomethyl ester (LP-13-3)

[0487] Compound P28 (0.627 g, 1.45 mmol), compound LP-13-2 (0.8 g, 1.45 mmol), and scandium trifluoromethanesulfonate (711 mg, 1.45 mmol) were added to 1,2-dichloroethane (15 mL). After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 3 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was concentrated to obtain a yellow solid compound LP-13-3 (0.8 g, crude). The yellow solid compound LP-13-3 was used directly in the next step. LCMS: (ESI, m / z): 928.2 [M+H] + .

[0488] Step 5: (2S)-2-[(2S)-2-(2-aminoacetamido)propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP-13-4)

[0489] Compound LP-13-3 (0.8 g, crude) and morpholine (1.26 g, 14.5 mmol) were added to N,N-dimethylformamide (5 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 250*70 mm*10 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-50% acetonitrile, retention 18 min) to obtain a white solid compound LP-13-4 (240 mg, yield 9.65%). LCMS: (ESI, m / z): 706 [M+H] + .

[0490] Step 6: (9H-fluorene-9-yl)methyl N-[2-(2-{[({[(1S)-1-{[(1S)-2-aminocarbonyl-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}ethyl]aminocarbonyl}methyl)aminocarbonyl]methoxy}ethoxy)ethyl]aminomethyl ester (LP-13-5)

[0491] At 25°C, LP-13-4 (230 mg, 325 μmol), 2,5-dioxoylide-1-yl-1-(9H-fluorene-9-yl)-3-oxoylide-2,7,10-trioxa-4-azadodecane-12-ester (157.25 mg, 325 μmol), and N,N-diisopropylethylamine (63.18 mg, 488 μmol) were added to N,N-dimethylformamide (4.6 mL). The reaction mixture was stirred at 25°C for 1 hour. After the reaction was complete, the LP-13-5 reaction solution was used directly for the next step. LCMS:(ESI) + ,m / z):1073.3[M+H] + .

[0492] Step 7: (2S)-2-[(2S)-2-(2-{2-[2-(2-aminoethoxy)ethoxy]acetamido}acetamido)propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP-13-6)

[0493] Morpholine (425.92 mg, 4.89 mmol) was added to the N,N-dimethylformamide (4.6 mL) reaction solution of compound LP-13-5 from the previous step. The reaction solution was stirred at 25 °C for 1 hour. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 100*40mm*50um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 8 min) to obtain a white solid compound LP-13-6 (110 mg, yield 15.47%). LCMS: (ESI, m / z): 851.3 [M+H] + .

[0494] Step 8: (2S)-2-[(2S)-2-[2-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)acetamido]propionylamino]-N-[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl]oxo)methyl]butanediamide (LP13)

[0495] LP-13-6 (110 mg, 50.42 μmol) was dissolved in N,N-dimethylformamide (2.4 mL), and bromoacetic acid succinimide ester (17.85 mg, 75.63 μmol) was added. The reaction mixture was stirred at 25 °C for 2 hours. After the reaction was complete, the reaction mixture was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Welch Xtimate C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 8 min) to obtain a white solid compound LP13 (23 mg, yield 46.94%). LCMS: (ESI, m / z): 971.2 / 973.2 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ8.99(s,1H),8.55(t,J=6.50Hz,1H),8.21-8.36(m,2H),8.17(d,J =7.75Hz,1H),8.00(d,J=10.63Hz,1H),7.85(t,J=5.63Hz,1H),7.27-7.43(m,2H),6.91(br s,1H),6.53(s,1H),5.43(s,2H),5.33(s,2H),4.69-4.86(m,2H),4.50-4 .63(m,3H),4.29(quin,J=7.07Hz,1H),3.91(s,2H),3.84(s,2H),3.80(br d,J=5.75Hz,2H),3.51-3.62(m,4H),3.39-3.45(m,2H),3.23(q,J=5.71Hz,3H),2. 56-2.65(m,4H),1.80-1.93(m,2H),1.23(d,J=7.00Hz,3H),0.88(t,J=7.38Hz,3H).

[0496] Example 43. (4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]butyric acid (LP14)

[0497] Step 1: Synthesis of 2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]acetic acid (LP-14-2)

[0498] Compound LP-14-1 (25 g, 47.84 mmol) was dissolved in anhydrous acetonitrile (500 mL). Ethyl di(propane-2-yl)amine (5.56 g, 43.06 mmol, 7.50 mL) was added to the system, followed by the slow addition of a 200 mL solution of 2-aminoacetic acid (3.95 g, 52.63 mmol) in water. After the reaction apparatus was purged three times, the reaction was carried out at 15 °C for 16 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 (250*100 mm, 15 μm); mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 50%-80% acetonitrile, retention 24 min) to obtain a white solid compound LP-14-2 (20 g, yield 86.6%). LCMS:(ESI,m / z):483.3[M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.39-12.86(m,1H)8.11-8.31(m,1H)7.89(d,J=7.38Hz,2H)7.74(br t,J=7.07Hz,2H)7.58(br d,J=8.38Hz,1H)7.37-7.48(m,2H)7.26-7.37(m,2H)4.16-4.35(m,3H)3.98-4.11(m,1H)3.64-3.88(m,2H)2.27(br t,J=7.88Hz,2H)1.85-1.97(m,1H)1.68-1.81(m,1H)1.39(s,9H).

[0499] Step 2: Synthesis of tert-butyl(4S)-4-({2-[(2,5-dioxoylidene-1-yl)oxo]-2-oxoylideneethyl}aminocarbonyl)-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester (LP-14-3)

[0500] Compound LP-14-2 (20 g, 41.45 mmol) was dissolved in anhydrous acetonitrile (400 mL), and N,N'-dicyclohexylmethyldiimide (9.41 g, 45.59 mmol, 9.22 mL) and 1-hydroxypyrrolidine-2,5-dione (5.25 g, 45.59 mmol) were added. After purging the reaction apparatus three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filter cake was washed once with acetonitrile (50 mL). The filtrate was concentrated to give a white solid compound LP-14-3 (23 g, yield 95.7%). The solid was used directly in the next step. LCMS: (ESI, m / z): 602.3 [M+Na]+ .

[0501] Step 3: Synthesis of (2S)-2-{2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]acetamido}-5-(aminocarbonylamino)valeric acid (LP-14-4)

[0502] Compound LP-14-3 (20 g, 34.51 mmol) was dissolved in anhydrous acetonitrile (400 mL). Ethyl di(propane-2-yl)amine (5.35 g, 41.41 mmol, 7.21 mL) and (2S)-2-amino-5-(aminocarbonylamino)valeric acid (7.25 g, 41.41 mmol) were added to the system. After the reaction apparatus was purged three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated by high performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 (250*100 mm, 15 μm); mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 20%-58% acetonitrile, retention for 20 min) to obtain a white solid compound LP-14-4 (18 g, yield 81.54%). 1 H NMR(400MHz, DMSO-d6)δ12.20-13.12(m,1H)8.15(br t,J=5.57Hz,1H)8.06(br d,J=7.75Hz,1H)7.89(d,J=7.50Hz,2H)7.73(t,J=7.44Hz,2H)7.60(br d,J=7.88Hz,1H)7.38-7.48(m,2H)7.27-7.37(m,2H)5.94(br t,J=5.57Hz,1H)5.39(s,2H)4.11-4.37(m,4H)3.96-4.06(m,1H)3.64-3.84(m,2H)2.93(q,J=6.46Hz,2H)2.25(br t,J=7.75Hz,2H)1.84-1.97(m,1H)1.64-1.80(m,2H)1.49-1.61(m,1H)1.39(s,11H).

[0503] Step 4: Synthesis of 2,5-dioxylidene-1-yl(2S)-2-{2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxylidenevalamido]acetamido}-5-(aminocarbonylamino)pentyl ester (LP-14-5)

[0504] Compound LP-4-4 (8 g, 12.51 mmol) was dissolved in anhydrous acetonitrile (160 mL), and N,N'-dicyclohexylmethyldiimide (2.84 g, 13.76 mmol, 2.78 mL) and 1-hydroxypyrrolidine-2,5-dione (1.58 g, 13.76 mmol) were added. After purging the reaction apparatus three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered and washed once with acetonitrile (50 mL). The filtrate was concentrated to give a white solid compound LP-14-5 (8 g, yield 86.82%), which was used directly in the next step. LCMS: (ESI, m / z): 737.2 [M+H] + .

[0505] Step 5: Synthesis of 2-[(2S)-2-{2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]acetamido}-5-(aminocarbonylamino)valamido]acetic acid (LP-14-6)

[0506] Compound LP-14-5 (8 g, 10.86 mmol) was dissolved in anhydrous acetonitrile (160 mL). Ethyldi(propane-2-yl)amine (1.68 g, 13.03 mmol, 2.27 mL) was added to the system. 2-Aminoacetic acid (978.11 mg, 13.03 mmol) was dissolved in water (40 mL) and slowly added dropwise to the reaction solution. After the reaction apparatus was purged three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*100mm*15um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP-14-6 (6.7 g, yield 88.56%). LCMS:(ESI,m / z):786.2[M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.34-12.77(m,1H)8.14-8.31(m,1H)8.10(br t,J=5.44Hz,1H)7.96(br d,J=8.00Hz,1H)7.89(d,J=7.50Hz,2H)7.73(t,J=7.57Hz,2H)7.58(br d,J=7.88Hz,1H)7.38-7.48(m,2H)7.27-7.37(m,2H)5.89(br t,J=5.38Hz,1H)5.36(s,2H)4.15-4.42(m,4H)3.96-4.07(m,1H)3.61-3.84(m,4H)2.93(br d,J=6.13Hz,2H)2.24(br t,J=7.75Hz,2H)1.83-1.99(m,1H)1.64-1.78(m,2H)1.49-1.58(m,2H)1.39(s,10H).

[0507] Step 6: Synthesis of tert-butyl(4S)-4-[({[(1S)-1-{[(acetoxy)methyl]aminocarbonyl}-4-(aminocarbonylamino)butyl]aminocarbonyl}methyl)aminocarbonyl]-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester (LP-14-7)

[0508] Compound LP-14-6 (1.5 g, 2.15 mmol) and copper acetate (273.72 mg, 1.51 mmol) were dissolved in N,N-dimethylformamide (30 mL). Lead tetraacetate (2.86 g, 6.46 mmol) was then added. After purging the reaction apparatus three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*100mm*15um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 45%-75% acetonitrile, retention 18 min) to obtain a white solid compound LP-14-7 (700 mg, yield 41.18%). LCMS: (ESI, m / z): 733.4 [M+Na] + . 1H NMR(400MHz, DMSO-d6)δ8.98(br t,J=6.75Hz,1H)8.12(br t,J=5.44Hz,1H)8.02(br d,J=7.75Hz,1H)7.89(d,J=7.50Hz,2H)7.73(br t,J=7.19Hz,2H)7.60(br d,J=7.88Hz,1H)7.38-7.46(m,2H)7.27-7.37(m,2H)5.92(br t,J=5.57Hz,1H)5.38(s,2H)5.07(br d,J=7.13Hz,2H)4.18-4.43(m,4H)3.94-4.07(m,1H)3.74(br d,J=5.63Hz,2H)2.85-2.99(m,2H)2.24(br t,J=7.63Hz,2H)1.95-2.04(m,3H)1.85-1.94(m,1H)1.69-1.79(m,1H)1.57-1.68(m,1H)1.25-1.52(m,12H).

[0509] Step 7: (4S)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyric acid (LP-14-8)

[0510] P28 (330 mg, 759.64 μmol) was dissolved in anhydrous 1,2-dichloroethane (33 mL), and scandium trifluoromethanesulfonate (147.39 mg, 759.64 μmol) and compound LP-14-7 (660.00 mg, 928.57 μmol) were added. After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 16 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was directly evaporated to dryness to obtain a yellow solid compound LP-14-8 (660 mg, crude product), which was used directly in the next step. LCMS: (ESI, m / z): 1029.3 [M+H] + .

[0511] Step 8: (4S)-4-amino-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]butyric acid (LP-14-9)

[0512] Compound LP-14-8 (660 mg, 641.38 μmol) was dissolved in N,N-dimethylformamide (13.2 mL) solvent, and morpholine (1.12 g, 12.83 mmol, 1.13 mL) was added. After the reaction apparatus was purged three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated by high performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100*30*10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-14-9 (150 mg, yield 28.99%). LCMS: (ESI, m / z): 807.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ12.03-12.57(m,1H)8.99-9.07(m,1H)8.96(br t,J=6.69Hz,1H)8.69(br t,J=5.44Hz,1H)8.23-8.37(m,1H)8.07-8.21(m,3H)8.01(d,J=10.76Hz,1H)7.35(s,1H)6.38-6.74(m,1H)5.98(br d,J=4.13Hz,1H)5.13-5.61(m,3H)4.71-4.88(m,2H)4.48-4.70(m,2H)4.18-4.39(m,1H)3.79-4.05(m,4H)2.96(br s,2H)2.61(br d,J=1.50Hz,3H)2.37-2.43(m,2H)1.80-2.00(m,4H)1.64-1.76(m,1H)1.55(td,J=8.57,4.75Hz,1H)1.30-1.46(m,2H)0.88(br t,J=7.19Hz,3H).

[0513] Step 9: (4S)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]-4-(2-{2-[2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)ethoxy]ethoxy}acetamido)butyric acid (LP-14-10)

[0514] Compound LP-14-9 (150 mg, 185.92 μmol) was dissolved in N,N-dimethylformamide (3 mL) solvent, and ethyldi(propane-2-yl)amine (28.83 mg, 223.11 μmol, 38.86 μL) and compound 1-12 (150 mg, 185.92 μmol) were added. After the reaction apparatus was purged three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was evaporated to dryness to obtain a brown liquid compound LP-14-10 (150 mg, crude). LCMS: (ESI, m / z): 1174.3 [M+H] + .

[0515] Step 10: (4S)-4-{2-[2-(2-aminoethoxy)ethoxy]acetamido}-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]butyric acid (LP-14-11)

[0516] Compound LP-14-10 (150 mg, 127.75 μmol) was dissolved in N,N-dimethylformamide (3 mL), and morpholine (222.59 mg, 2.55 mmol, 224.84 μL) was added. The reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100*30*10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-14-11 (70 mg, yield 57.56%). LCMS: (ESI, m / z): 952.2 [M+H] + . 1H NMR(400MHz, DMSO-d6)δ11.99-12.29(m,1H)9.02(s,1H)8.83-8.95(m,1H)8.30-8.42(m,1H)8.10(br d,J=8.00Hz,1H)8.01(br d,J=10.76Hz,1H)7.62-7.87(m,4H)7.36(s,1H)6.54(br s,1H)5.97(br s,1H)5.23-5.53(m,5H)4.67-4.91(m,2H)4.52-4.64(m,2H)4.21-4.44(m,2H)3.89-4.01(m,2H)3.80(br d,J=5.38Hz,2H)3.61(br s,6H)2.89-3.05(m,4H)2.61(s,3H)2.26(br t,J=7.88Hz,2H)1.76-1.99(m,4H)1.65-1.75(m,1H)1.49-1.61(m,1H)1.31-1.45(m,2H)0.88(br t,J=7.19Hz,3H).

[0517] Step 11: (4S)-4-(2-{2-[2-(2-bromoacetamido)ethoxy]ethoxy}acetamido)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]butyric acid (LP14)

[0518] Compound LP-14-11 (70 mg, 73.53 μmol) and 2,5-dioxoylide-1-yl-2-bromoacetate (173.55 mg, 735.33 μmol) were dissolved in N,N-dimethylformamide (1.4 mL), and 4-methylmorpholine (7.44 mg, 73.53 μmol) was added. The reaction was carried out at 15 °C for 0.5 h under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 15%-45% acetonitrile, retention 8 min) to obtain a yellow solid compound LP14 (30 mg, yield 38.03%). LCMS: (ESI, m / z): 1072.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.11(s,1H)9.02(s,1H)8.81-8.95(m,1H)8.23-8.41(m,2H)8.09(br d,J=7.63Hz,1H)8.00(d,J=10.51Hz,1H)7.78(br d,J=7.75Hz,1H)7.35(s,1H)6.55(s,1H)5.94(br t,J=5.44Hz,1H)5.26-5.52(m,6H)4.68-4.91(m,2H)4.59(s,2H)4.20-4.40(m,2H)3.92(s,2H)3.85(s,2H)3.79(br d,J=5.38Hz,2H)3.49-3.65(m,4H)3.38-3.48(m,2H)3.19-3.28(m,2H)2.87-3.03(m,2H)2.56-2.65(m,3H)2.26(br t,J=7.75Hz,2H)1.75-1.99(m,4H)1.64-1.74(m,1H)1.55(br dd,J=9.13,4.88Hz,1H)1.30-1.46(m,2H)0.88(t,J=7.25Hz,3H).

[0519] Example 44. (4S)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 ,20 Synthesis of 2-(2,5-dioxane-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]-4-[2-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)acetamido]butyric acid (LP15)

[0520] Step 1: Synthesis of 2,5-dioxylidene-1-yl(2S)-2-{2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxylidenevalamido]acetamido}-5-(aminocarbonylamino)pentyl ester (LP-15-2)

[0521] Compound LP-15-1 (8 g, 12.51 mmol) was dissolved in anhydrous acetonitrile (160 mL), and N,N'-dicyclohexylmethyldiimide (2.84 g, 13.76 mmol, 2.78 mL) and 1-hydroxypyrrolidine-2,5-dione (1.58 g, 13.76 mmol) were added. After purging the reaction apparatus three times, the reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed once with acetonitrile (50 mL). The filtrates were combined and concentrated to obtain a white solid compound LP-15-2 (8 g, yield 86.8%), which was used directly in the next reaction. LCMS: (ESI, m / z): 737.2 [M+H] + .

[0522] Step 2: Synthesis of 2-[(2S)-2-{2-[(2S)-5-(tert-butoxy)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-5-oxoylidenevalamido]acetamido}-5-(aminocarbonylamino)valamido]acetic acid (LP-15-3)

[0523] Compound LP-15-2 (8 g, 10.86 mmol) was dissolved in anhydrous acetonitrile (160 mL). Diisopropylethylamine (1.68 g, 13.03 mmol, 2.27 mL) and 2-aminoacetic acid (978.11 mg, 13.03 mmol) in water (40 mL) were added to the system, respectively. The reaction was carried out at 15 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*100mm*15um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP-15-3 (6.7 g, yield 88.5%). LCMS: (ESI, m / z): 697.3 [M+H] + .

[0524] Step 3: Synthesis of (tert-butyl(4S)-4-[({[(1S)-1-{[(acetoxy)methyl]aminocarbonyl}-4-(aminocarbonylamino)butyl]aminocarbonyl}methyl)aminocarbonyl]-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyl ester)(LP-15-4)

[0525] Compound LP-15-3 (4 g, 5.74 mmol) and copper acetate (729.92 mg, 4.02 mmol) were added to N,N-dimethylformamide (80 mL). Lead tetraacetate (7.64 g, 17.22 mmol) was then added. After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 2 hours under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 250*100mm*15um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 40%-55% acetonitrile, retention 21 min) to obtain a white solid compound LP-15-4 (0.96 g, yield 23.5%). LCMS: (ESI) + ,m / z):711.3[M+H] + .

[0526] Step 4: (4S)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 ,20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]-4-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)butyric acid)(LP-15-5)

[0527] P28 (0.3 g, 690.58 μmol) was dissolved in anhydrous 1,2-dichloroethane (30 mL), and scandium trifluoromethanesulfonate (339.88 mg, 690.58 μmol) and compound LP-15-4 (736.27 mg, 1.04 mmol) were added. After purging the reaction apparatus three times, the reaction was carried out at 60 °C for 12 hours under a nitrogen atmosphere. After the reaction was complete, the yellow solid compound LP-15-5 (600 mg, crude product) obtained by rotary evaporation was directly used in the next step of the reaction. LCMS:(ESI) + ,m / z):1029.0[M+H] + .

[0528] Step 5: ((4S)-4-amino-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]) 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]butyric acid)(LP-15-6)

[0529] Compound LP-15-5 (0.8 g, 777.44 μmol) was dissolved in N,N-dimethylformamide (8 mL) solvent, and morpholine (677.28 mg, 7.76 mmol) was added. After the reaction apparatus was purged three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. Adding 0.8 mL of formic acid to the filtrate resulted in the precipitation of some impurities. The filtrate after filtration was subjected to preparative high-performance liquid chromatography (HPLC) (column type: WePure Biotech XP tC18 100*30*10 μm; mobile phase [water (0.1% trifluoroacetic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a yellow solid compound LP-15-6 (100 mg, yield 12.75%). LCMS: (ESI) + ,m / z):807.3[M+H] + .

[0530] Step 6: (4S)-4-[({[(1S)-4-(aminocarbonylamino)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 2-(2,5-dioxane-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}butyl]aminocarbonyl}methyl)aminocarbonyl]-4-[2-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)acetamido]butyric acid (LP15)

[0531] Compound LP-15-6 (15 mg, 18.59 μmol) and maleimide succinimide ester (14.07 mg, 55.78 μmol) were added to N,N-dimethylformamide (0.3 mL). N,N-diisopropylethylamine (1.20 mg, 9.30 μmol) was added under a nitrogen atmosphere at 0 °C. After purging the reaction apparatus three times, the reaction was carried out at 0 °C for 30 minutes under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered, and the filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 10%-40% acetonitrile, retention 8 minutes) to obtain a yellow solid compound LP15 (5.8 mg, yield 31.56%). LCMS: (ESI, m / z): 944.4 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.01(s,1H),8.85(br t,J=6.63Hz,1H),8.39-8.48(m,1H),8.23-8.31(m,1H),8.04-8.11(m,1H),7.99(d,J=10. 63Hz,1H),7.35(s,1H),7.06(s,2H),6.54(s,1H),5.93-6.00(m,1H),5.43(s,2H),5.39(br s,2H),5.33(s,2H),4.73-4.85(m,2H),4.58(s,2H),4.24-4.29(m,2H),4.04-4.15(m,3H),3.78(br d,J=5.25Hz,1H),2.87-3.01(m,2H),2.60(d,J=1.38Hz,3H),2.25(br t,J=7.94Hz,2H),1.81-1.97(m,3H),1.64-1.79(m,2H),1.47-1.59(m,1H),1.33-1.45(m,2H),0.87(t,J=7.32Hz,3H). 19 F NMR(377MHz,DMSO-d6)δ110.36.

[0532] Example 45. (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0]2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP16)

[0533] Step 1: (9H-fluorene-9-yl)methyl N-[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminomethyl ester (LP-16-1)

[0534] Compound P28 (350 mg, 805.68 μmol), compound 1-11 (581.95 mg, 1.21 mmol), and 4-methylbenzenesulfonic acid (55.50 mg, 322.27 μmol) were dissolved in N,N-dimethylformamide (7.0 mL). The reaction was carried out at 40 °C for 12 hours under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was used directly for the next reaction. LCMS: (ESI, m / z): 856.3 [M+H] + .

[0535] Step 2: (2S)-2-amino-N-[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]-3-methylbutyramide (LP-16-2)

[0536] Compound LP-16-1 (crude product ~350 mg, 408.90 μmol) and morpholine (356.25 mg, 4.08 mmol, 359.85 μL) were dissolved in N,N-dimethylformamide (7 mL). The reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100*30 mm*5 μm; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-35% acetonitrile, retention 8 min) to obtain a white solid compound LP-16-2 (180 mg, yield 65.7%). LCMS: (ESI, m / z): 634.3 [M+H] + .

[0537] Step 3: Methyl(4S)-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}-4-[(2S)-2-({[(9H-fluorene-9-yl)methoxy]carbonyl}amino)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]butyl ester (LP-16-3)

[0538] Compound LP-16-2 (180 mg, 284.06 μmol), compounds 1-9 (191.93 mg, 284.06 μmol), and 2-hydroxypyridine-1-cation-1-ol anion (78.90 mg, 710.15 μmol) were dissolved in N,N-dimethylformamide (1.8 mL), followed by the addition of ({[3-(dimethylamino)propyl]aminoethylenedimethyl)methyl(ethyl)amine (167.70 mg, 710.15 μmol, 191.22 μL, HBr salt). The reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered. The filtrate was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex Luna C18 100*30mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 25%-55% acetonitrile, retention 8 min) to give a white solid compound LP-16-3 (112.5 mg, yield 30.6%). LCMS: (ESI, m / z): 1291.6 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.95(s,1H)8.78(br t,J=6.82Hz,1H)8.18(br d,J=6.75Hz,1H)7.97-8.07(m,2H)7.87(br d,J=7.50Hz,2H)7.81(br d,J=8.38Hz,1H)7.68-7.76(m,3H)7.58(br d,J=7.63Hz,1H)7.37-7.44(m,2H)7.29-7.36(m,3H)6.54(s,1H)5.43(s,2H)5.31(s,2H)4.69-4.85(m,3H)4.58(s,2H)4.46(d,J=5 .50Hz,1H)4.38(d,J=6.00Hz,1H)4.14-4.36(m,8H)3.92-4.02(m,1H)3.51-3.64(m,5H)3.44-3.50(m,1H)3.35-3.43(m,2H)3.26(br s,2H)2.96-3.07(m,1H)2.60(d,J=1.50Hz,3H)2.30(br d,J=6.25Hz,2H)2.09-2.23(m,2H)1.68-2.03(m,7H)1.25(br d,J=7.00Hz,3H)0.70-0.93(m,9H).

[0539] Step 4: (4S)-4-[(2S)-2-amino-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20 Synthesis of 1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP-16-4)

[0540] Under a nitrogen atmosphere at 0°C, lithium hydroxide monohydrate (1M, 914.76 μL) was added to a mixed solvent of tetrahydrofuran (1.0 mL) and water (0.5 mL) containing LP-16-3 (112.5 mg, 87.12 μmol). The reaction was carried out at 25°C for 0.5 h under a nitrogen atmosphere. After the reaction was complete, the reaction solution was filtered. The filtrate was separated by preparative high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*3um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-35% acetonitrile, retention 8 min) to obtain a white solid compound LP-16-4 (30 mg, yield 32.64%). LCMS: (ESI, m / z): 1055.6 [M+H] + .

[0541] Step 5: (4S)-4-[(2S)-2-(2-bromoacetamido)-4-{[(2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl]aminocarbonyl}butyrylamino]-4-{[(1S)-1-{[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] 2 , 11 .0 4 , 9 .0 15 , 20Synthesis of 21-carbon-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]prop-2-yn-1-yl}oxo)methyl]aminocarbonyl}ethyl]aminocarbonyl}-2-methylpropyl]aminocarbonyl}butyric acid (LP16)

[0542] Compound LP-16-4 (30 mg, 28.43 μmol) and 2,5-dioxonylpyrrolidone-1-yl 2-bromoacetate (33.56 mg, 142.17 μmol) were dissolved in N,N-dimethylformamide (0.6 mL). After purging the reaction apparatus three times, the reaction was carried out at 25 °C for 1 hour under a nitrogen atmosphere. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated by high-performance liquid chromatography (HPLC) (column type: Phenomenex luna C18 100*40mm*5um; mobile phase [water (0.2% formic acid)-acetonitrile]; mobile phase: 5%-45% acetonitrile, retention 8 min) to obtain a white solid compound LP16 (13 mg, yield 39%). LCMS: (ESI, m / z): 1177.2 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ12.09(s,1H)8.96(s,1H)8.77(br t,J=6.63Hz,1H)8.48(d,J=7.75Hz,1H)8.18(br t,J=7.88Hz,2H)8.00(d,J=10.63Hz,1H)7.68-7.82(m,2H)7.35(s,1H)6.53(s,1H)5.43(s,2H)5.33 (s,2H)4.71-4.84(m,3H)4.59(s,2H)4.45(d,J=5.63Hz,1H)4.37(d,J=6.00Hz,1H)4.22-4.34(m,5H) 4.18(dd,J=8.19,6.94Hz,1H)3.84-3.95(m,2H)3.53-3.64(m,3H)3.43-3.50(m,1H)3.35-3.42(m,2 H)3.22-3.28(m,1H)2.97-3.06(m,1H)2.61(d,J=1.63Hz,3H)2.24(td,J=10.13,6.13Hz,2H)2.12(br t,J=7.82Hz,2H)1.97(br dd,J=13.63,6.75Hz,1H)1.87(tt,J=14.70,7.13Hz,4H)1.66-1.79(m,2H)1.26(d,J=7.13Hz,3H)0.80-0.91(m,9H). 1H NMR(400MHz, DMSO-d6+D2O)δ8.93(s,1H)8.74(br t,J=6.75Hz,1H)8.51(d,J=7.50Hz,1H)8.19(br t,J=7.00Hz,2H)7.91(d,J=10.51Hz,1H)7.80(br d,J=7.50Hz,2H)7.37(s,1H)5.33-5.46(m,2H)5.28(s,2H)4.76(br d,J=6.50Hz,2H)4.54(s,2H)4.13-4.28(m,3H)4.06-4.12(m,1H)3.80-3.86(m,2H)3.50-3.62(m,3H)3.43 -3.49(m,1H)3.33-3.41(m,2H)3.19-3.29(m,1H)2.95-3.04(m,1H)2.56(s,3H)2.17-2.31(m,2H)2.11(br t,J=7.50Hz,2H)1.92-2.00(m,1H)1.85(dq,J=14.70,7.28Hz,4H)1.67-1.79(m,2H)1.24(br d,J=7.00Hz,3H)0.76-0.88(m,9H).

[0543] Synthesis of antibody-drug conjugates (ADCs):

[0544] Example 46. Synthesis of trastuzumab-LP1-DAR4:

[0545] Step 1: Antibody Preparation

[0546] 452 mg of lyophilized trastuzumab powder was dissolved in 22 mL of pure water. The obtained antibody was dialyzed for 4 cycles using an ultrafiltration tube (30 KD) with 50 mM buffer solution (pH = 7.4, containing 6.86 g disodium hydrogen phosphate dihydrate and 1.58 g disodium hydrogen phosphate monohydrate per liter) to obtain an antibody solution of 8.63 mg / mL (using the trastuzumab extinction coefficient ε). 280 =213380M -1 cm -1 ).

[0547] Step 2: Antibody reduction

[0548] To a solution containing 12.2 mL (105 mg, 0.000724 mmol) of trastuzumab, add 6.2 mL of 50 mM buffer solution (pH = 7.4, containing 6.86 g disodium hydrogen phosphate dihydrate and 1.58 g sodium dihydrogen phosphate monohydrate per liter), then add 144.8 μL of 10 mM tris(2-carboxyethyl)phosphine solution (TCEP) and 2.1 mL of a mixed solution of 10 mM diethylenetriaminepentaacetic acid (DTPA) and sodium hydroxide (containing 3.90 g DTPA and 1.20 g sodium hydroxide per liter). Place the reaction mixture in a multi-functional mixer and incubate at 25 °C for 2 hours.

[0549] Step 3: Coupling between antibody and linker-payload

[0550] Add 6-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)-N-{[({[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] to the trastuzumab reducing solution obtained in step 2. 2 , 11 .0 4 , 9 .0 15 , 20 A solution of 2,1-C-1(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]aminocarbonyl}-2-phenylethyl]aminocarbonyl}methyl)aminocarbonyl]methyl}hexamamide (LP1, 0.00441 mmol) in dimethyl sulfoxide (0.88 mL). The reaction solution was placed in a multi-functional mixer and reacted at 25 °C for 1 hour.

[0551] Step 4: Purification

[0552] The reaction solution obtained in step 3 was purified by 6 cycles using an ultrafiltration tube (30KD) with 25mM buffer (pH=5.5, containing 0.90g L-histidine and 4.04g L-histidine hydrochloride monohydrate per liter) to obtain a solution of trastuzumab-LP1-DAR4. Size exclusion chromatography (SEC) showed a monomer content of 98.2%. Hydrophobic chromatography (HIC) showed a drug-to-antibody ratio (DAR) of 4.0.

[0553] Example 47. Synthesis of trastuzumab-LP1-DAR8:

[0554] Step 1: Antibody Preparation

[0555] 452 mg of lyophilized trastuzumab powder was dissolved in 22 mL of pure water. The obtained antibody was dialyzed for 4 cycles using an ultrafiltration tube (30 KD) with 50 mM buffer solution (pH = 7.4, containing 6.86 g disodium hydrogen phosphate dihydrate and 1.58 g disodium hydrogen phosphate monohydrate per liter) to obtain an antibody solution of 8.63 mg / mL (using the trastuzumab extinction coefficient ε). 280 =213380M -1 cm -1 ).

[0556] Step 2: Antibody reduction

[0557] To a solution containing 12.2 mL (105 mg, 0.000724 mmol) of trastuzumab, add 6.2 mL of 50 mM buffer solution (pH = 7.4, containing 6.86 g disodium hydrogen phosphate dihydrate and 1.58 g sodium dihydrogen phosphate monohydrate per liter), then add 579.2 μL of 10 mM tris(2-carboxyethyl)phosphine solution (TCEP) and 2.1 mL of a mixed solution of 10 mM diethylenetriaminepentaacetic acid (DTPA) and sodium hydroxide (containing 3.90 g DTPA and 1.20 g sodium hydroxide per liter). Place the reaction mixture in a multi-functional mixer and incubate at 25 °C for 2 hours.

[0558] Step 3: Coupling between antibody and linker-payload

[0559] Add 6-(2,5-dioxane-2,5-dihydro-1H-pyrrolo-1-yl)-N-{[({[(1S)-1-{[({3-[(19S)-19-ethyl-6-fluoro-19-hydroxy-7-methyl-14,18-dioxane-17-oxa-3,13-diazapentacyclo[11.8.0.0] to the trastuzumab reducing solution obtained in step 2. 2 , 11 .0 4 , 9 .0 15 , 20 A solution of 1,76 mL dimethyl sulfoxide (LP1, 0.00882 mmol) of 2,1-(21),2,4,6,8,10,15(20)-heptaen-8-yl]phenyl}aminocarbonyl)methyl]aminocarbonyl}-2-phenylethyl]aminocarbonyl}methyl)aminocarbonyl]methyl}hexamamide (LP1, 0.00882 mmol) was prepared. The reaction solution was placed in a multi-functional mixer and reacted at 25 °C for 1 hour.

[0560] Step 4: Purification

[0561] The reaction solution obtained in step 3 was purified by 6 cycles using an ultrafiltration tube (30KD) with 25mM buffer (pH=5.5, containing 0.90g L-histidine and 4.04g L-histidine hydrochloride monohydrate per liter) to obtain a solution of trastuzumab-LP1-DAR8. Hydrophobic chromatography (HIC) showed a drug-antibody ratio (DAR) of 7.7.

[0562] Example 48. Synthesis of other antibody-drug conjugates:

[0563] Referring to the experimental protocols of Example 34 (preparation of DAR4-ADC) and Example 35 (preparation of DAR8-ADC), other antibody-drug conjugates were prepared by changing the corresponding conditions as described in Table 1 below.

[0564] Table 1. Antibody-drug conjugates and their preparation methods

[0565] Bioactivity evaluation:

[0566] The control compounds used in the bioactivity evaluation section were the marketed drug DS-8201 (CAS: 1826843-81-5) and the payload Dxd of DS-8201 (CAS: 1599440-33-1). MB-3a (CN 115551552 A) was additionally added as a control compound in some bioactivity evaluation sections. MB-3a is an antibody-drug conjugate; the antibody portion is trastuzumab, using Cys-based random conjugation, with a DAR value of 8. The linker-toxin portion is MB-3, and its structure is as follows:

[0567] Experimental Example 1. In vitro cell activity assay of toxins

[0568] Cells in the logarithmic growth phase were digested and counted. Cell suspensions of 3000 cells per well were evenly seeded onto 96-well plates (100 μL per well). The test compound was diluted at different concentrations to prepare a series of 3× dilutions. 50 μL of each serially diluted sample was added to each well of the 96-well plate, gently vortexed, and incubated in a CO2 incubator (37°C, 5% CO2) for 120 hours. Cell Counting Lite 2.0 (Vazyme) reagent was removed from a -20°C freezer and brought to room temperature in the dark. 50 μL of the reagent was added to each well, and the plates were incubated at 300 rpm for 10 min in the dark. Luminescence signal was detected using a microplate reader. A dose-inhibition curve was generated using GraphPad Prism software, and the IC50 was calculated. 50 value.

[0569] The results of the in vitro cell activity assays of the toxins are summarized in Table 2 below. The results show that the toxins disclosed herein all exhibit strong proliferative inhibitory activity in tumor cells of different indications, with several toxins showing significantly better activity than the control molecule Dxd, indicating greater application potential.

[0570] Table 2 Results of in vitro cytotoxicity assays for toxins

[0571] Experimental Example 2. In vitro liver microsomal stability study of toxins

[0572] According to the "Technical Guidelines for Non-Clinical Studies of Antibody-Drug Conjugates" (No. 46, 2023), "Although ADCs increase the targeted release of small molecule compounds, some small molecule compounds are still released prematurely. Some free small molecule compounds can also rapidly diffuse or be transported from target cells to surrounding tissues and systemic circulation, or apoptosis or damage to target cells may lead to free small molecule compounds entering systemic circulation." The guidelines further state that "ADCs differ from general antibody drugs; their toxicity primarily stems from free small molecule compounds." Current industry consensus holds that, from a metabolic stability perspective, an ideal payload, once entering the circulatory system, needs to be rapidly metabolized to reduce systemic exposure and systemic toxicity. Therefore, we evaluated the hepatic microsomal stability of toxins from some examples to determine their suitability as ideal payloads.

[0573] Experimental Methods: Human liver microsomes (final concentration 0.5 mg / mL), the analyte (final concentration 1 μM), and 100 mM phosphate buffer were added to an incubation plate. The plate was preheated at 37 °C for 5 min. NADPH was added to the test wells, and the plate was incubated in a 37 °C water bath. Samples were taken at 0, 5, 15, 30, and 60 min, and stop reaction solution (200 μL, solvents: 100 μL methanol and 100 μL acetonitrile, solutes: 50 ng / mL labetalol and 50 ng / mL tolbutamide) was added. The samples were centrifuged at 4000 rpm for 10 min. 50 μL of the supernatant was diluted with 150 μL of pure water and analyzed by LC-MS.

[0574] The half-life is calculated using the following formula: T 1 / 2 = (-0.693 / k), where k is the slope of the scavenging curve, which is a fitted straight line. The horizontal axis represents the incubation time, and the vertical axis represents the natural logarithm of the concentration of the analyte.

[0575] The clearance rate is calculated using the following formula: CL int =(0.693 / T) 1 / 2 T*(1 / incubation concentration of liver microsomes)*scale factor. Where, T1 / 2 The half-life is indicated by the incubation concentration of liver microsomes, which is 0.5 mg / mL, and the scale factor is 900.

[0576] Experimental results: As shown in Table 3, some compounds in this invention have higher clearance rates compared to Dxd, suggesting that these compounds may be more ideal payloads than Dxd.

[0577] Table 3. Results of in vitro liver microsomal stability of the toxin

[0578] Experimental Example 3. Study on the inhibitory activity of toxins on topoisomerase I

[0579] Prepare the reaction buffer as follows: Buffer (2×): 20 mM Tris-HCl, pH 7.5, 100 mM KCl, 10 mM MgCl2, 30 μg / mL BSA, 0.5 mM DTT

[0580] Conduct the experiment following these steps:

[0581] 1: Take 5 μL Buffer (2×), add 0.5 μg supercoiled DNA, 1 U recombinant human TOP1 protein, and the compound to be detected, and bring the total volume to 10 μL with ddH2O. Mix gently.

[0582] 2: React at 37℃ for 30 minutes;

[0583] 3: Add 2 μL of 6×Loading buffer to the reaction solution, and use the prepared 0.7% agarose gel in 1×TAE buffer for electrophoresis at a constant voltage of 50V for 1 h;

[0584] 4: Remove the agarose gel after electrophoresis, soak it in a staining solution containing 1×gel red for 30 minutes, and take pictures with a UV gel camera for observation.

[0585] Table 4. Results of the inhibitory activity of toxins on topoisomerase I

[0586] The tested examples showed good topoisomerase I inhibitory activity.

[0587] Experimental Example 4. In vitro cytotoxicity assay of ADC

[0588] NCI-N87 and SK-BR-3 cells in logarithmic growth phase were digested and counted. Cell suspensions of 3000 cells per well were evenly seeded onto 96-well plates (100 μL per well). ADC was diluted to prepare a series of 3× concentrations. 50 μL of each serially diluted ADC sample was added to each well of the 96-well plate, gently vortexed, and incubated in a CO2 incubator (37°C, 5% CO2) for 120 hours. Cell Counting Lite 2.0 (vazyme) reagent was removed from a -20°C freezer and brought to room temperature in the dark. 50 μL of the reagent was added to each well, and the plates were incubated at 300 rpm for 10 min in the dark. Luminescence signal was detected using a microplate reader. A dose-inhibition curve was generated using GraphPad Prism software, and the IC50 was calculated. 50 value.

[0589] Table 5 Results of ADC in vitro cytotoxicity assay

[0590] "-" indicates that it has not been tested.

[0591] The in vitro cell-killing results of ADC molecules are shown in Table 5. For most of the embodiments, the IC... 50 Both the maximum kill ratio and the IC in some embodiments are no weaker than DS-8201 and MB-3a. 50 The maximum kill ratio was better than that of the positive control.

[0592] Experimental Example 5. Study on the toxin release efficiency of ADC

[0593] Following the experimental protocol for liver microsome stability, the ADC to be tested was incubated with cathepsin B or liver lysosomes. Samples were collected at 6 time points within 24 hours, and the samples were quantitatively analyzed by LC-MS to test the content of the target toxin in order to evaluate the toxin release efficiency of the ADC.

[0594] Experimental Example 6. Plasma Stability Study of ADCs

[0595] Following the experimental protocol for liver microsomal stability, plasma was incubated with the ADC to be tested. Samples were collected at 6 time points within 24 hours, and LC-MS was used to quantitatively analyze the samples to test the content of the target toxin in order to evaluate the plasma stability of the ADC.

[0596] Experimental Example 7. In vitro activity assay of ADC bystander effect

[0597] Bystander cells (with low antigen expression, such as MDA-MB-468 cells with low HER2 expression) were digested, centrifuged, and washed once with PBS. The CFSE stock solution was diluted 1:5000 with PBS, and the bystander cells were resuspended in the diluted CFSE stock solution. The cells were then labeled and placed in a CO2 incubator for 15 minutes. Subsequently, the cells were washed twice with DMEM complete medium containing 10% FBS and counted, adjusting the cell density to 6 × 10⁶ cells / year. 5 cells / mL. Following the experimental design, seed cells into 6-well plates, 1 ml per well. Target cells (high antigen expression, such as NCI-N87 with high HER2 expression) were digested, centrifuged, and counted, adjusting the cell density to 2 × 10⁶ cells / mL. 5 cells / ml. Following the experimental design, 1 mL of cells was seeded into each well of a 6-well plate. The ADC sample stock solution was diluted with RPMI Medium 1640 + 10% FBS to a drug concentration (2 × analytical concentration) of 20 nM. 1 mL of the diluted sample stock solution was added to each well of the plate according to the experimental design. An untreated group was also established, in which 2 mL of RPMI Medium 1640 + 10% FBS was added to each well of a 6-well plate. After mixing the target cells, observer cells, and sample, the plates were incubated in a CO2 incubator for 5 days.

[0598] After 5 days of incubation, the 6-well plates were removed. On the one hand, the supernatant of each group of cells was collected and digested, centrifuged and counted. On the other hand, the cells were collected using flow cytometry to detect the percentage of CFSE-positive cells, and the raw data were analyzed using FlowJo. Finally, based on the number of target cells and bystander cells remaining in each group, the data were analyzed using GraphPadPrism software.

[0599] Table 6. Cell killing by the bystander effect on antigen-low expression of Raji-Luc

[0600] In this invention, the bystander effect of Trastuzumab-LP14 is superior to that of DS-8201 and MB-3a.

[0601] Experimental Example 8. In vivo efficacy of ADC in the NCI-N87 CDX model

[0602] Female Balb / c-Nude mice were subcutaneously inoculated with NCI-N87 cells in the right abdomen at a concentration of 5 × 10⁶ cells. 6 / animal, inoculated with 0.2 mL (containing 50% Matrigel). On day 6 post-inoculation, the average tumor volume reached 160 mm. 3At approximately 10:00 AM, the animals were randomly divided into groups of 8 mice each. The tumor-bearing mice were treated with different administration regimens via tail vein injection of the test drug and the control drug.

[0603] Tumor volume and mouse body weight were measured twice weekly, and T / C (%) and TGI (%) were calculated. Tumor volume V (in mm) 3 The calculation formula is: V = 0.5a × b 2 , where a and b are the long and short dimensions of the tumor, respectively. Results are expressed as mean and standard error (mean ± SEM). All data were analyzed using Graphpad Prism, and p < 0.05 was considered statistically significant. The relative tumor proliferation rate T / C (%) was calculated as: T / C (%) = (Ti / T0) / (Vi / V0) × 100%. The relative tumor growth inhibition rate TGI (%) was calculated as: TGI (%) = [1 - (Ti / T0) / (Vi / V0)] × 100%. Ti refers to the mean tumor volume of the treatment group measured at each specified time point after treatment; T0 refers to the tumor volume of the treatment group at the time of grouping; Vi refers to the mean tumor volume of the mediator control group measured at each specified time point after treatment; V0 refers to the tumor volume of the mediator control group at the time of grouping. If T / C > 40%, there is no efficacy; if T / C = < 40% and p < 0.05, tumor inhibition exists.

[0604] Table 7. TGI of ADC in NCI-N87 CDX model (Day 21)

[0605] The test results are shown in Table 7. All the tested ADC molecules showed ideal in vivo efficacy in the NCI-N87 model. Among them, Trastuzumab-LP14 showed the best activity, which was superior to the positive control DS-8201.

[0606] Table 8. TGI of ADC in NCI-N87 CDX model (Day 28 and Day 42)

[0607] The results of long-term in vivo efficacy evaluation are shown in Table 8. Both Trastuzumab-LP14 and Trastuzumab-LP16 showed potent and long-lasting antitumor activity, which was significantly better than the positive control DS-8201.

[0608] Experimental Example 9. In vivo efficacy of ADC in the JIMT-1CDX model

[0609] Scid-Beige mice were subcutaneously inoculated with JIMT-1 cells in the right ventricular region at a cell density of 1 × 10⁻⁶ cells. 7 / animal, inoculated with 0.2 mL (containing 50% Matrigel). On day 6 post-inoculation, the average tumor volume reached 175 mm. 3 At approximately 10:00 AM, the animals were randomly divided into groups of 8 mice each. The tumor-bearing mice were treated with different administration regimens via tail vein injection of the test drug and the control drug.

[0610] Table 9. TGI of ADC in JIMT-1 CDX model (Day 21)

[0611] The test results are shown in Table 9. All the tested ADC molecules exhibited ideal in vivo efficacy in the JIMT-1 model. Among them, the novel molecules disclosed in this patent showed significantly stronger in vivo activity than the marketed drug DS-8201.

Claims

1. A compound of Formula (I) or a pharmaceutically acceptable salt thereof; wherein R 1 and R 2 are the same or different and independently hydrogen, hydroxyl, thiol, halogen, cyano, -NR c R d , C 1-6 1-6 alkyl, -(CH2CH2O) m R c , C 1-6 1-6 alkoxy, C 1-6 1-6 alkylthio, -C(=O)NR c R d , -NR c C(=O)R d , -SOR c , or -SO2R c , said C 1-6 1-6 alkyl, -(CH2CH2O) m R c , C 1-6 1-6 alkoxy, C 1-6 1-6 alkylthio, -C(=O)NR c R d , -NR c C(=O)R d , -SOR c , and -SO2R c are optionally substituted with halogen, hydroxyl, or amino; or R 1 and R 2 together form a ring of formula -X-L a -Y-; X and Y are each independently selected from -NH- or -S-; L a is a C 1-3 alkylene or C 1-3 haloalkylene; Alk is C 2-6 alkylene, C 2-6 alkynylene, C 5-8 cycloalkylene, C 6-10 aryl, 5-10 membered heterocycloalkylene, or 5-10 membered heteroarylene, said C 2-6 alkylene, C 2-6 alkynylene, C 5-8 cycloalkylene, C 6-10 aryl, 5-10 membered heterocycloalkylene, and 5-10 membered heteroarylene are optionally substituted with 1-8 R a substituents; R a independently halogen, cyano, nitro, hydroxy, mercapto, oxo (=0), -NR c R d , carboxy, -C(=0)NR c R d , -N(R c )C(=0)R d , -SO2NR c R d , -N(R c )SO2R d , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 haloalkyl, -(CH2CH2O) m R c , C 3-8 cycloalkyl, 3-8 membered heterocycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio, -SOR c , -SO2R c , -P(=0)R c R d , -NR c C(=0)NR c R d or -C(=0)R c ; Q 1 is absent, -NH-, -O-, -S-, -C(=O)-, -C(=O)NH-, -NHC(=O)-, -C(=O)O-, -OC(=O)-, -S(=O)-, -SO2-, -SO2NH-, -NHSO2-, -S(=O)-NH-, -NH-S(=O)-, -P(=O)R c -, -NHS(=O)NH-, -NHC(=O)NH-, -NHC(=S)NH- or -NHC(=NH)NH-; L and L b independently absent, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, -(CH2CH20) m -, -OC 1-6 alkylene, -N(C 1-6 alkyl)-, -S-C 1-6 alkylene, C 3-8 cycloalkylene or 3-8 membered heterocycloalkylene; said C 1-6 alkylene, C 2-6 alkenylene, C 2- alkynylene, -(CH2CH20) m -, -OC 1-6 alkylene, -N(C 1-6 alkyl)-, -S-C 1-6 alkylene, C 3-8 cycloalkylene and 3-8 membered heterocycloalkylene are optionally substituted with halogen or C 3-8 cycloalkyl; and L b is absent, Q 1 is absent; m is independently an integer from 1 to 10; Q 2 is hydrogen, hydroxy, mercapto or amino; the amino group being optionally substituted by C 1-6 alkyl, C 3-8 cycloalkyl, C 2-6 alkenyl or C 2-6 alkynyl; R c and R d independently hydrogen or C 1-6 alkyl; the heteroatoms in the 5-10 membered heterocyclenyl, 5-10 membered heteroarylenyl and 3-8 membered heterocyclalkylenyl are one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula (I), wherein the compound of Formula (I) satisfies one or more of the following conditions: (1) R 1 and R 2 are the same or different, and independently are hydrogen, hydroxyl, halogen, cyano, C 1-6 alkyl, C 1-6 haloalkyl, -NR c R d , thiol, C 1-6 alkoxy, C 1-6 alkylthio, -S(=O)R c , or -SO2R c ; wherein R c is C 1-6 alkyl; R d is hydrogen or C 1-6 alkyl; (2) Alk is C 2-6 imidene group, C 5-8 Cycloalkylene, C 2-6 Alynyl, phenylene, 5-6 membered heterocyclic alkenyl or 5-6 membered heteroaryl, wherein C 2-6 imidene group, C 5-8 Cycloalkylene, C 2-6 The alkynyl, phenylene, 5-6 membered heterocyclic alkenyl or 5-6 membered heteroaryl group is optionally surrounded by 1-8 R groups. a Substitution; the heteroatoms in the 5-6 membered heterocyclic alkenyl and 5-6 membered heteroaryl are one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3; (3) R a independently halogen, cyano, oxo (=0), C 1-6 alkyl, C 1-6 alkoxy or C 1-6 haloalkyl; (4) Q 1 is absent, -NH-, -O-, b -NHC(=O)- or b -C(=O)NH-, b is attached to L; (5) L and L b independently absent, C 1-6 alkylene, C 1-6 haloalkylene, or C 3-8 cycloalkyl-substituted C 1-6 alkylene; (6) Q 2 is hydrogen, hydroxy, amino or amino substituted by C 1-6 alkyl.

3. The compound of claim 2, or a pharmaceutically acceptable salt thereof, having the formula (I), wherein the compound of Formula (I) satisfies one or more of the following conditions: (1)R 1 -CH3; (2) R 2 is -F; (2) Alk is a is attached to L; (4) Q 1 is absent or b -NHC(=0)-, b is connected to L; (5) L is absent; (6) L b is absent, -CH2- or -CH2-CH2-; (7) Q 2 is hydrogen, hydroxy, amino or methylamino.

4. The compound of claim 2, or a pharmaceutically acceptable salt thereof, having the formula (I), wherein R 1 is C 1-6 alkyl; R 2 is halogen; Alk is C 2-6 alkynylene, C 5-8 cycloalkenylene, C 6-10 aryl, 5-10 membered heterocycloalkenylene, or 5-10 membered heteroaryl, said C 2- alkynylene, C 5-8 cycloalkenylene, C 6-10 aryl, 5-10 membered heterocycloalkenylene, and 5-10 membered heteroaryl are optionally substituted with 1-2 R a substituents; R a independently halogen, cyano, C 1-6 alkyl, C 1-6 alkoxy or C 1-6 haloalkyl; Q 1 is absent, -NH-, -O-, b -NHC(=O)- or b -C(=O)NH-, b is attached to L; L is absent; L b is absent or C 1-6 alkylene; and L b Q is absent when 1 is absent; Q 2 is hydrogen, hydroxy, amino or amino substituted by C 1-6 alkyl.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the formula (I), wherein the compound of Formula (I) is any one of the compounds in Table A.

6. A compound of Formula (II) or a pharmaceutically acceptable salt thereof, L 1 -D (II) wherein, L 1 are linker precursors; D is a group formed by the loss of a hydrogen atom from the compound of Formula (I) according to any one of claims 1-5.

7. The compound of formula (II) as claimed in claim 6, or a pharmaceutically acceptable salt thereof, characterized in that, L 1 for MM 1 -L c -C(=O)-AA-SE-; wherein MM 1 is hydrogen or an electrophilic group; L c is absent, -(CH2) q -, c -(CH2) q1 C(=O)N(R e )(CH2) q2 -, c -(CH2) q1 (OCH2CH2) q2 -, c -(CH2) q1 (OCH2CH2) q2 N(R e )C(=O)(CH2) q1 -, c -(CH2) q3 C(=O)N(R e )(CH2CH2O) q4 (CH2) q5 - or c -(CH2CH2O) q6 (CH 2) q7 - is attached to the C-terminus of MM 1 ; R e is hydrogen or C 1-6 alkyl; q and q1 are independently an integer from 1 to 10; q2 is an integer from 1 to 100; q3, q4, q5, q6 and q7 are independently an integer from 1 to 10; AA is a polypeptide residue formed from 2-10 amino acids, which are the same or different, and optionally substituted with one or more polyols or C 1-6 alkyl; the C-terminus of the polypeptide residue is connected to SE, and the N-terminus is connected to -C(=0)-; SE is absent, 1 is attached to AA and 2 is attached to D; R 3 is hydrogen or -L d -Q 3 -L e -MM 2 ; L d is absent, C 1-10 alkylene or C 3-10 cycloalkylene; Q 3 -NR f -, -O-, -S-, -C(=O)-, -N(R f )(C=O)-, -C(=O)N(R f )-, -OC(=O)-, -C(=O)O-, -S(=O)-, -SO2-, -SO2NR f -, -NR f SO2-, -S(=NH)(=O)-, -P(=O)R f -, -NHC(=O)NH-, -NHC(=S)NH- or -NHC(=NH)NH-; L e is absent, -(CH2) p -, d -(CH2) p1 C(=O)N(R g )(CH2) p2 -, d -(CH2) p1 (OCH2CH2) p2 N(R g )C(=O)(CH2) p1 -, d -(CH2) p1 C(=O)N(R g )CH2CH2(OCH2CH2) p2 - or d -(CH2) p1 (OCH2CH2) p2 - is covalently attached to the MM 2 at the d terminus; R f and R g are independently hydrogen or C 1-6 alkyl; p and p1 are independently an integer from 1 to 10; p2 is an integer from 1 to 100; MM 2 is an electrophilic group; R 3 with MM 1 only one is hydrogen; D is Q 2a is absent, -O-, -NH- or -N(C 1-6 alkyl)-; R 1 , R 2 , Alk, L, Q 1 and L b are defined as in any one of claims 1 to 5.

8. The compound of claim 7, or a pharmaceutically acceptable salt thereof, having the formula (II), wherein, the compound of Formula (II) satisfies one or more of the following conditions: (1) MM 1 is hydrogen, halogen, a Michael acceptor group, an epoxy group, R h independently hydrogen or C 1-6 alkyl; (2) L c -(CH2) q -, c -CH2C(=O)NHCH2CH2-, c -CH2CH2OCH2CH2OCH2CH2NHC(=O)CH2CH2-, c -(CH2) q3 C(=O)NH(CH2CH2O) q4 (CH2) q5 - or c -(CH2CH2O)2-CH2-, c-terminal to MM 1 is connected; q is 1-6; q3, q4, and q5 are independently 1, 2, 3, 4, 5, or 6; (3) AA is a polypeptide residue formed from 2, 3 or 4 amino acids, which are the same or different, and optionally substituted by one or more or C 1-6 alkyl substitution; the C-terminus of the polypeptide residue is connected to SE and the N-terminus is connected to -C(=0)-; preferably -Val-Ala-, -Ala-Val-, -Gly-Gly-, -Val-Cit-, -Cit-Val-, -Glu-Ala-, -Glu-Lys-, -Glu-Gly-, -Glu-Cit-, -Leu-Ala-, -Ala-Leu-, -Leu-Cit-, -Cit-Leu-, -Lys-Lys-, -Ala-Lys-, -Lys-Ala-, -Val-Lys-, -Lys-Val-, -Tyr-Arg-, -Arg-Tyr-, -Arg-Arg-, -Ala-Ala-, -Phe-Lys-, -Lys-Phe-, -Thr-Thr-, -Thr-Met-, -Met-Thr-, -Met-Tyr-, -Tyr-Met-, -Phe-Gln-, -Gln-Phe-, -Gly-Ser-, -Leu-Gln-, -Gln-Leu-, -Ser-Ala-, -Ser-Gly-, -Val-Thr-, -Thr-Val-, -Val-Gln-, -Ser-Val-, -Val-Ser-, -Ala-Met-, -Met-Ala-, -Val-Arg-, -Arg-Val-, -Phe-Ala-, -Ala-Phe-, -Gln-Val-, -Phe-Arg-, -Arg-Phe-, -Ala-Ala-Ala-, -Glu-Gly-Cit-, -Gly-Gly-Gly-, -Ala-Val-Ala-, -Gly-Val-Gly-, -Ala-Val-Gly-, -Gly-Phe-Lys-, -Lys-Phe-Gly-, -Leu-Ala-Leu-, -Val-Ala-Leu-, -Leu-Val-Ala-, -Val-Ala-Val-, -Gly-Lys-Val-, -Gly-Lys-Gly-, -Val-Lys-Gly-, -Glu-Val-Cit-, -Gln-Val-Cit-, -Glu-Val-Ala-, -Gln-Val-Ala-, -Glu-Lys-Gly-, -Glu-Phe-Gly-, -Ala-Val-Ala-Gly-, -Gly-Phe-Gly-Gly-, -Gly-Gly-Phe-Gly-, -Ala-Val-Gly-Gly-, -Gly-Gly-Val-Ala, -Ala-Ala-Ala-Ala-, -Ala-Leu-Ala-Leu-, -Leu-Ala-Leu-Ala-, -Gly-Phe-Leu-Gly-,-Gly-Leu-Phe-Gly-, -Glu-Glu-Lys-Gly-, -Glu-Glu-Val-Cit-, -Gln-Glu-Val-Cit-, -Glu-Glu-Val-Ala-, -Gln-Glu-Val-Ala-, -Gly-Glu-Lys-Gly-, -Gly-Glu-Phe-Gly-, -Gln-Gly-Phe-Gly-, -Glu-Glu-Gly-Cit-, -Ala-Ala-Asn-, Gly-Ala-Asn-, the C-terminus of AA is attached to SE and the N-terminus is attached to -C(=O)-; (4) L d is absent or C 1-6 alkylene; (5) Q 3 -NH(C=O)- e , -C(=O)NH- e or -C(=O)O- e , e is attached to L e ; (6) L e -(CH2) p - or d -(CH2) p1 C(=O)NH(CH2) p2 -, the d end is covalently linked to MM 2 p and pi are independently integers from 1 to 6, and p2 is independently an integer from 1 to 6; (7) MM 2 hydrogen, halogen, Michael acceptor groups, epoxy groups, R j independently hydrogen or C 1-6 alkyl.

9. The compound of formula (II) as claimed in claim 7, characterized in that, the compound of Formula (II) satisfies one or more of the following conditions: (1) MM 1 is hydrogen, -Br or (2) L c is -CH2-, -CH2-CH2-, -CH2-CH2-CH2-CH2-CH2-, c -CH2C(=0)NHCH2CH2OCH2CH2OCH2- or c -(CH2CH20)2-CH2-, c-terminally to MM 1 is attached; (3) AA is the C-terminal of AA is attached to SE and the N-terminus is attached to -C(=O)- (4) SE is absent, 1 is attached to AA and 2 is attached to D; (5) L d is not present; (6) Q 3 -C(=O)NH- e , e is connected to L e ; (7) L e is -(CH2)6- or d -CH2-C(=O)NH(CH2)2-, d end to MM 2 covalently linked; (8) MM 2 is -Br or (9) Q 2a is absent, -O-, -NH-, or -N(CH3)-.

10. The compound of formula (II) as claimed in claim 7, characterized in that, D is Q 2a is absent, -O-, -NH- or -N(C 1-6 alkyl)-; L 1 for MM 1 -L c -C(=O)-AA-SE-; MM 1 halogen or L c -(CH2) q -, q is 1-6; AA is a polypeptide residue formed from 2, 3 or 4 amino acids, which are the same or different, and optionally substituted by one or more or C 1-6 alkyl substituted; the C-terminus of the polypeptide residue is connected to SE and the N-terminus is connected to -C(=0)-; SE is absent, 1 is attached to AA and 2 is attached to D; R 3 is hydrogen or -L d -Q 3 -L e -MM 2 ; L d is not present; Q 3 -NH(C=O)- e , -C(=O)NH- e or -C(=O)O- e , e is attached to L e ; L e -(CH2) p -, d -(CH2) p1 C(=O)NH(CH2) p2 -,d end covalently linked to MM 2 ; p and p1 are integers independently from 1 to 6; p2 is an integer from 1 to 10; MM 2 R is hydrogen, halogen or R 3 with MM 1 only one is hydrogen; R 1 , R 2 , Alk, L, Q 1 , and L b are as defined in any one of claims 1-5.

11. The compound of claim 7, or a pharmaceutically acceptable salt thereof, having the formula (II), ###0002### (II) the compound of Formula (II) is any one of the compounds in Table B.

12. A conjugate of Formula (III) or a pharmaceutically acceptable salt thereof; Ab-[L 2 -D]p (III) wherein, Ab is an antibody or an antigen-binding fragment thereof; L 2 is a linker; p is the drug loading and is any value between 1 and 20; D is defined as in any one of claims 6-11.

13. The conjugate of Formula (III) or a pharmaceutically acceptable salt thereof according to claim 12, wherein, the conjugate of Formula (III) satisfies one or more of the following conditions: (1) L 2 for f -MM 3 -L c -C(=O)-AA-SE- or ​ f is attached to Ab; SE is absent, 1 is attached to AA and 2 is attached to D; MM 1 R is hydrogen; R 3a -L d -Q 3 -L e -MM 4 - f , f end is attached to Ab; MM 3 and MM 4 is independently absent or f is attached to Ab; L c , AA, L d , Q 3 , and L e are as defined in any one of claims 7-11. (2) Ab is an antibody or an antigen-binding fragment thereof targeting HER2, HER3, HER4, EGFR, B7H3, TROP2, CD19, CD79b, CD33, FOLR1 or Nectin4; (3) p is any value between 1 and 10.

14. The conjugate of Formula (III) or a pharmaceutically acceptable salt thereof according to claim 13, wherein, the conjugate of Formula (III) satisfies one or more of the following conditions: (1) Ab is anbenitamab, coprelotamab, disitamab, gancotamab, margetuximab, timigutuzumab, zanidatamab, trastuzumab, pertuzumab, inetetamab or an antigen-binding fragment thereof; (2) Ab is demupitamab, depatuxizumab, futuximab, imgatuzumab, laprituximab, losatuxizumab, matuzumab, modotuximab, necitumumab, nimotuzumab, panitumumab, pimurutamab, serclutamab, tomuzotuximab, zalutumumab, Cetuximab, intetumumab, laprituximab, or an antigen-binding fragment thereof; (3) Ab is enoblituzumab, mirzotamab, omburtamab, ifinatamab, vobramitamab antibody, or an antigen-binding fragment thereof; (4) Ab is datopotamab, sacituzumab, or an antigen-binding fragment thereof; (5) p is 1, 2, 3, 4, 5, 6, 7, 8, or a fraction between 2 and 9.

15. The conjugate of claim 12, represented by Formula (III): ###00010### or a pharmaceutically acceptable salt thereof. The conjugate of formula (III) is any of the conjugates in Table C.

16. The conjugate of Formula (III) or a pharmaceutically acceptable salt thereof according to claim 12, wherein, The conjugate of Formula (III) is any one of the following:

17. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1-5, or a conjugate of formula (III) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 12-16, and at least one pharmaceutical excipient.

18. Use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any of claims 1-5, a conjugate of formula (III) or a pharmaceutically acceptable salt thereof as claimed in claims 12-16, or a pharmaceutical composition as claimed in claim 17, for the manufacture of a medicament for the prevention or treatment of a disease; preferably a disease associated with abnormal cell activity, such as esophageal cancer, brain tumor, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, pancreatic cancer, breast cancer, head and neck cancer, cervical cancer, endometrial cancer, colorectal cancer, liver cancer, kidney cancer, urothelial cancer, solid tumor, non-Hodgkin's lymphoma, leukemia, central nervous system tumor, prostate cancer, thyroid cancer, or large B-cell lymphoma.

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