Polypeptide conjugate compound, obtained polypeptide conjugate drug, and use thereof

Abstract

The present application provides a polypeptide conjugate compound consisting of a homing peptide, a payload, and a linker. The obtained polypeptide conjugate compound contains a triazine ring linker capable of selectively labeling a phenolic hydroxyl group of tyrosine or a thiol group of cysteine, or contains a maleimide linker capable of selectively labeling a thiol group of cysteine. A drug developed therefrom can solve the problems of poor plasma stability, low release rate, insufficient pharmacokinetics, etc. of such drugs, and is effectively used in the treatment and / or prevention of diseases.

Description

Polypeptide conjugate compounds, resulting polypeptide conjugate drugs and uses thereof

[0001] The present application claims priority to the Chinese patent application No. 2024112705219, filed on September 11, 2024, and entitled "Polypeptide conjugate compounds, resulting polypeptide conjugate drugs and uses thereof", the entire content of which is incorporated herein by reference. TECHNICAL FIELD

[0002] The present application belongs to the technical field of biological medicine, and in particular relates to a polypeptide conjugate compound, a resulting polypeptide conjugate drug and uses thereof. BACKGROUND

[0003] Tumors are still the main cause of death in humans. Small molecule chemotherapy drugs are prone to produce serious toxic side effects due to their poor targeting and biological distribution, leading to poor patient compliance and difficulty in obtaining satisfactory clinical benefits. Polypeptide conjugate drugs (PDC) have good targeting and pharmacokinetic properties due to their selectivity for tumor cells, and can overcome the shortcomings of ordinary chemotherapy drugs, and have become a new hotspot in the research and development of anti-tumor drugs in recent years.

[0004] PDCs for tumor treatment generally include three components: homing peptides, linkers and payloads. The three parts cooperatively deliver chemotherapy drugs through receptors on tumor cells to expand their therapeutic effect. Polypeptides have better tissue penetration, lower manufacturing cost, higher potency, activity, selectivity and specificity, fewer drug interactions and lower off-target toxicity, etc.

[0005] In the development process of polypeptide conjugate drugs, there are many challenges and problems. The polypeptide molecules are easily affected by enzyme digestion, oxidation, polymerization and other factors in the body, leading to loss of activity or toxicity; the linker plays a bridge role between the polypeptide and the drug molecule (i.e. the payload), and the linker affects the plasma stability, release rate and pharmacokinetics of PDC, and the labeling position of the linker on the polypeptide affects the targeting of the drug. At present, most of the polypeptide conjugate drugs are designed and synthesized based on functional groups such as sulfhydryl or phenolic hydroxyl in amino acids, and by adjusting the type and length of the linker, polypeptide conjugate drugs with appropriate stability, release rate and high anti-tumor activity are obtained. SUMMARY

[0006] In order to improve some characteristics of existing polypeptide conjugate drugs, the present application provides a polypeptide conjugate compound, a resulting polypeptide conjugate drug and uses thereof; the polypeptide conjugate compound comprises a homing peptide, a linker and a payload.

[0007] The first aspect of the present application provides a polypeptide conjugate compound, whose structural formula is shown in formula (I):

[0008] P is a homing peptide, optionally a transmembrane peptide or a targeting peptide; specifically, it can be selected from any one of the following polypeptides: c(RGDfC), SSTR, SSTR-1, CHN-1, 2YHN-1 and 2CHN-1. Among them, the c(RGDfC) represents a cyclic RGD polypeptide; the SSTR represents an Octreotide polypeptide; the SSTR-1 represents a (c(fCYwKTCTC)) polypeptide; the CHN-1 represents a CTSPLNIHNGQKL polypeptide; the 2YHN-1 represents a YGTSPLNIHNGQKLGY polypeptide; and the 2CHN-1 represents a CGTSPLNIHNGQKLGC polypeptide. The objects represented are polypeptides connected by amino acids.

[0009] D is a load; it can be selected from any one of the following groups: Plinabulin, sea slug toxin, auristatin, maytansine, MMAE (Monomethyl auristatin E), MMAF (Monomethylauristatin F), calicheamicin, duocarmycin, pyrrolobenzodiazepine, irinotecan, irinotecan hydrochloride, exatecan derivative, camptothecin, 7-ethyl-10-hydroxy camptothecin, 10,11-methylenedioxy camptothecin, 10,11-methylenedioxy-7-(3-furan) camptothecin, 10,11-methylenedioxy-7-(3-thiophene) camptothecin, 10,11-methylenedioxy-7-(1-(3-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)amino)methyl) camptothecin, and 10,11-methylenedioxy-7-(1-(3-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)amino)methyl) camptothecin.

[0010] In formula (I), the part between P (homing peptide) and D (load) is a linker T-B-Z-L, and D and T-B-Z-L can constitute an intermediate, represented as T-B-Z-L-D.

[0011] T is wherein * is connected to P and # is connected to B; X is selected from any one of O, S, NH;

[0012] R' in the T is selected from any one of the following groups: -(CH2) n1 -, -(CH2CH2O) n2 -, C3-C8 carbocyclyl, phenyl, and heterocyclyl containing N, S or O, wherein n1 and n2 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11;

[0013] Y in T is selected from any one of the following group: H, N, S, CH2, and O;

[0014] R" in T is selected from any one of the following group: alkyl with carbon number 1-5, phenyl, naphthyl, halogen-substituted phenyl, styryl, methyl-substituted phenyl, alkyl alkyne with carbon number 1-10, and alkyl azide with carbon number 1-10.

[0015] B is selected from any one of the following group: -(CH2) n3 -, -(CH2CH2O) n4 -, -(CH2) n5 -S-S-(CH2) n6 -, C3-C8 carbocyclic group, and phenyl, wherein n3, n4, n5, n6 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11.

[0016] Z is -(AA) n7 , wherein AA is an amino acid or a polypeptide, and n7 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In one embodiment, AA is selected from any one of the following amino acid or peptide sequence: Val-Cit, Val-Ala, Val-Lys, Phe-Lys, Lys-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp, Lys, Cit, Phe-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Leu-Ala-Leu, Ile-Ala-Leu, Val-Ala-Val, Ala-Leu-Ala-Leu, β-Ala-Leu-Ala-Leu, Gly-Phe-Leu-Gly, and Gly-Gly-Phe-Gly.

[0017] m is selected from 0 or 1.

[0018] L is R is selected from any one of the following group: hydrogen, halogen, methyl, ethyl, nitro, methoxy, and ethoxy; q is the number of R q , which can be independently 0, 1, 2, 3, or 4; for example, R q may be nothing (q = 0) or one (q = 1), or a chain of multiple same or different groups, for example, R q a chain of halogen and ethyl (q = 2), R qa chain consisting of three methyl groups (q = 3), and so on. Likewise, * is connected to P and # is connected to B.

[0019] p is selected from 0 or 1.

[0020] In one embodiment, the more specific structural formula of each polypeptide in P is as follows:

[0021] The letters in each of the above-mentioned structural formula of each polypeptide in each of the above-mentioned structural formula of each polypeptide represent 1 amino acid, which is part of the polypeptide, for example, Y represents tyrosine, C represents cysteine, and N represents asparagine, which is well known to those skilled in the art.

[0022] In one embodiment, T-B-Z-L-D (i.e., intermediate) in the structural formula of the polypeptide conjugate compound is selected from any one of a1-a21; wherein the specific structure of a1-a21 is detailed in each compound obtained by reaction in Embodiment 1. In one embodiment, the polypeptide conjugate compound is selected from any one of b1-b27:

[0023] In one embodiment, the polypeptide conjugate compound can be at least extended to stereoisomers of the polypeptide conjugate compound, pharmaceutically acceptable salts of the polypeptide conjugate compound and its stereoisomers, and solvates of the polypeptide conjugate compound and its stereoisomers.

[0024] The present application also provides a polypeptide conjugate drug prepared from the polypeptide conjugate compound of any one of the preceding embodiments. It can be understood that in the preparation process, the polypeptide conjugate compound, the stereoisomer of the polypeptide conjugate compound, the pharmaceutically acceptable salt of the polypeptide conjugate compound and its stereoisomers, the solvate of the polypeptide conjugate compound and its stereoisomers, etc. can be selected to prepare the drug.

[0025] The present application also provides a use of the polypeptide conjugate compound of any one of the preceding embodiments for the preparation of a medicament for preventing or treating a disease or disorder, or reducing the severity of the disease or disorder.

[0026] The present application also provides a use of the polypeptide conjugate compound of any one of the preceding embodiments for preventing or treating a disease or disorder, or reducing the severity of the disease or disorder.

[0027] In an embodiment, the disease or disorder is cancer selected from at least one of the group consisting of breast cancer, squamous cell carcinoma, lung cancer, peritoneal cancer, liver cancer, gastric cancer, gastrointestinal cancer, membranous adenocarcinoma, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, urethral cancer, hepatocellular carcinoma, intestinal cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, melanoma, multiple myeloma, lymphoma, lymphoid tumor, brain cancer, gallbladder cancer, esophageal cancer, bile duct cancer, head and neck cancer, and metastatic tumors of the aforementioned cancers.

[0028] In an embodiment, the lung cancer is adenocarcinoma of the lung, squamous cell carcinoma of the lung, small cell lung cancer or non-small cell lung cancer; the breast cancer is HER2-positive breast cancer; the lymphoma is B-cell lymphoma; the squamous cell carcinoma is squamous epithelial cell carcinoma or oral squamous cell carcinoma.

[0029] In an embodiment, the disease or disorder is an infectious disease selected from at least one of the group consisting of respiratory infectious disease, digestive infectious disease, contact infectious disease, insect-borne infectious disease, blood-borne infectious disease, and body fluid-borne infectious disease.

[0030] Compared with the prior art, the application has the advantages and positive effects that:

[0031] In at least one embodiment of the application, the linker in the polypeptide conjugated drug designed and synthesized by the application contains a triazine ring, which can selectively label tyrosine phenolic hydroxyl or cysteine sulfhydryl, or the linker contains a maleimide, which can selectively label cysteine sulfhydryl.

[0032] The polypeptide conjugated compound and the polypeptide conjugated drug described in at least one embodiment of the application are evaluated in terms of plasma stability, in-vivo and in-vitro anti-tumor activity, and safety, and it is confirmed that the polypeptide conjugated drug designed and synthesized by the application has good plasma stability, good in-vivo and in-vitro anti-tumor activity, and good safety, which can solve the problems of poor plasma stability, low release rate, and insufficient pharmacokinetics of such drugs, and is effectively used in the treatment and / or prevention of diseases. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is the in-vivo anti-tumor evaluation result of compound b3 provided in an embodiment of the application in A549 xenograft mice (n = 6), wherein (A) is a tumor graph; (B) is a tumor weight; (C) is a tumor inhibition rate; (D) is the structural formula of compound b3; (E) is a tumor volume curve graph;

[0034] Figure 2 is the HE staining (200x magnification) detection result of the main organs of the nude mice after treatment with compound b3 provided in an embodiment of the application;

[0035] Figure 3 is a result of a comparison study of the in-vitro plasma stability of compound b3 and c(RGDfC)-MAL-FL118 according to an embodiment of the present application;

[0036] Figure 4 is a graph of the pharmacokinetic curve of compound b3 and c(RGDfC)-MAL-FL118 in rats according to an embodiment of the present application. DETAILED DESCRIPTION

[0037] The technical solutions in the embodiments of the present application will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0038] Example 1 Synthesis of linker-cytotoxin conjugate intermediate (T-B-Z-L-D):

[0039] This embodiment aims to prepare the desired linker-cytotoxin conjugate intermediate, and the linker is connected with the cytotoxin drug through a click reaction, etc. The synthesis is as follows.

[0040] 1. Preparation of compound 10-SN38-PABC-C6-Tri(a1)

[0041] In a 100 mL reaction bottle, compound 10-SN38-PABC-C6-N3 (200 mg, 0.29 mmol), 2-alkynyl butoxy-4-chloro-6-ethoxy-1, 3, 5-triazine (132 mg, 0.58 mmol), CuSO4·5H2O (363 mg, 1.45 mmol), sodium ascorbate (574 mg, 2.9 mmol) and TBTA (Tris-(benzyltriazolylmethyl) amine) (15 mg, 0.029 mmol) were added, acetonitrile and water (3:1, 12 mL) were added, and the reaction was stirred at room temperature for 12 h. After TLC (Thin-layer chromatography) detection showed that the reaction was complete, 50 mL of water was added to quench the reaction, and the reaction liquid was extracted with EA (Ethyl acetate) (30 mL x 3) three times. The extracted organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. Finally, the organic phase was filtered, vacuum concentrated, and purified by silica gel (PE:EA = 10:1-1:2) (PE is petroleum ether) column chromatography to obtain white solid 10-SN38-PABC-C6-Tri (179 mg, 68%). 1H NMR (400 MHz, DMSO-d6) δ 10.02 (s, 1H), 8.23 (d, J = 9.2 Hz, 1H), 8.16 (d, J = 2.4 Hz, 1H), 7.97 (s, 1H), 7.78 (dd, J = 9.2, 2.4 Hz, 1H), 7.65 (d, J = 8.5 Hz, 2H), 7.42 (d, J = 8.5 Hz, 2H), 7.34 (s, 1H), 5.43 (s, 2H), 5.34 (s, 2H), 5.26 (s, 2H), 4.61 (t, J = 6.6 Hz, 2H), 4.43 - 4.39 (m, 2H), 4.36 (s, 2H), 3.23 - 3.17 (m, 2H), 3.11 (t, J = 6.6 Hz, 3H), 2.31 (t, J = 7.4 Hz, 2H), 2.05 - 1.96 (m, 2H), 1.85 (dd, J = 9.3, 5.1 Hz, 2H), 1.64 (p, J = 8.0, 7.1 Hz, 4H), 1.29 (dd, J = 12.7, 5.0 Hz, 9H), 0.95 - 0.80 (m, 6H). HRMS: m / z [M+H] + C 45 H 46 ClN9O 10 ,908.3056; found, 908.3057.

[0042] 2. Preparation of compound SN38-PABC-C6-Tri(a2)

[0043] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-C6-N3, to give white solid 204 mg, yield 65%. 1H NMR (400 MHz, DMSO-d6) δ 8.02 (d, J = 7.5 Hz, 1H), 7.71 - 7.63 (m, 3H), 7.53 (d, J = 2.2 Hz, 1H), 7.43 - 7.37 (m, 2H), 7.34 (dd, J = 7.5, 1.5 Hz, 1H), 7.19 (d, J = 1.4 Hz, 1H), 5.70 - 5.59 (m, 2H), 5.25 - 5.14 (m, 2H), 5.10 - 4.93 (m, 3H), 4.85 (td, J = 12.2, 2.7 Hz, 1H), 4.63 - 4.39 (m, 3H), 4.32 (td, J = 12.2, 3.3 Hz, 1H), 3.66 - 3.54 (m, 1H), 3.04 - 2.88 (m, 2H), 2.88 - 2.76 (m, 1H), 2.71 (td, J = 12.0, 2.7 Hz, 1H), 2.59 - 2.40 (m, 2H), 2.32 (dq, J = 12.5, 7.9 Hz, 1H), 1.79 - 1.61 (m, 2H), 1.63 - 1.43 (m, 3H), 1.31 (dt, J = 24.0, 8.1 Hz, 6H), 0.94 (t, J = 8.0 Hz, 3H). HRMS: m / z [M + H] + C 45 H 46 C 10 ,908.3056; found, 908.3257.

[0044] 3. Preparation of compound SN38-8PEG-Tri(a3)

[0045] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-8PEG-N3, to give 244 mg of white solid in 51% yield. 1H NMR (400 MHz, DMSO-d6) δ 7.37 (dd, J = 7.5, 1.5 Hz, 1H), 7.19 (d, J = 1.5 Hz, 1H), 5.80 (dd, J = 9.5, 1.0 Hz, 1H), 5.18 - 5.10 (m, 2H), 5.02 - 4.80 (m, 4H), 4.65 - 4.45 (m, 3H), 4.37 - 4.19 (m, 3H), 3.93 (ddd, J = 12.4, 9.0, 1.2 Hz, 1H), 3.70 (dt, J = 12.6, 1.7 Hz, 1H), 3.67 - 3.58 (m, 2H), 3.58 (dt, J = 3.7, 2.0 Hz, 2H), 3.59 - 3.53 (m, 2H), 3.56 - 3.45 (m, 2H), 3.44 - 3.21 (m, 8H), 3.18 - 2.85 (m, 13H), 2.64 - 2.40 (m, 3H), 1.27 (dt, J = 16.3, 8.0 Hz, 6H), 0.89 (t, J = 8.0 Hz, 3H). HRMS: m / z [M+H] + C 48 H 61 ClN8O 16 , 1041.3894; found, 1041.3895.

[0046] 4. Preparation of compound FL118-S-S-Tri(a4):

[0047] In a 50 mL single-necked flask, 2-((4-chloro-1,3,5-triazin-2-yl)amino)ethane-1- thiol (62 mg, 0.32 mmol) and FL118-S-S (100 mg, 0.16 mmol) were added, 10 mL THF (Tetrahydrofuran) was added to dissolve, and the reaction was stirred at room temperature for 12 h. TLC detection showed that the reaction was complete. 50 mL H2O was added, and DCM (Dichloromethane) was extracted twice. The organic phase was combined, dried over anhydrous MgSO4, filtered, and the filtrate was concentrated under reduced pressure. Column chromatography gave 98 mg of a light yellow solid, with a yield of 87%. 1H NMR (400 MHz, DMSO-d6) δ 8.43 (s, 1H), 8.02 (s, 1H), 7.98 (s, 1H), 7.46 (d, J = 9.8 Hz, 2H), 6.95 (d, J = 2.9 Hz, 1H), 6.26 (d, J = 4.4 Hz, 2H), 5.49 (s, 2H), 5.21 (d, J = 3.8 Hz, 2H), 4.23 (dd, J = 11.2, 6.0 Hz, 1H), 4.18 - 4.12 (m, 1H), 2.92 - 2.87 (m, 2H), 2.79 (t, J = 7.2 Hz, 1H), 2.43 (d, J = 9.3 Hz, 2H), 2.20 (m, J = 6.8 Hz, 2H), 1.23 (d, J = 9.1 Hz, 3H), 0.90 (t, J = 8.4 Hz, 3H). 13 C NMR (125 MHz, DMSO-d6) δ 172.44, 165.41, 157.18, 154.84, 151.84, 149.18, 147.07, 130.63, 105.23, 103.05, 78.46, 72.86, 60.81, 43.97, 36.97, 35.13, 23.06, 21.51, 19.08, 17.00, 8.01. MS (ESI) m / z: 699.10 [M+H] + .

[0048] 5. Preparation of compound 10-SN38-PABC-K-C6-Tri(a5)

[0049] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to 10-SN38-PABC-K-C6-N3, to give white solid 214 mg, yield 67%. 1 H NMR (400 MHz, DMSO-d6) δ 1H NMR (500 MHz, Chloroform-d) δ 9.50 (s, 1H), 8.06 (d, J = 7.5 Hz, 1H), 7.74-7.66 (m, 3H), 7.50 (s, 1H), 7.48-7.42 (m, 2H), 7.40-7.33 (m, 2H), 5.77 (dd, J = 9.4, 0.8 Hz, 1H), 5.63 (dd, J = 9.5, 1.0 Hz, 1H), 5.15 (dd, J = 17.7, 1.0 Hz, 1H), 4.95 (dd, J = 17.7, 1.0 Hz, 1H), 4.85 (t, J = 7.0 Hz, 1H), 4.67-4.55 (m, 2H), 4.38-4.25 (m, 2H), 4.12-3.97 (m, 2H), 3.94 (s, 1H), 3.42 (dq, J = 12.4, 8.0 Hz, 1H), 3.10-2.98 (m, 2H), 2.90-2.79 (m, 3H), 2.53 (td, J = 12.3, 1.2 Hz, 1H), 2.43 (dq, J = 12.5, 8.0 Hz, 1H), 2.21 (tdd, J = 12.2, 5.7, 2.4 Hz, 1H), 2.01-1.69 (m, 5H), 1.65-1.48 (m, 3H), 1.46-1.34 (m, 1H), 1.38-1.24 (m, 9H), 0.88 (t, J = 8.0 Hz, 4H), 0.76 (dddd, J = 13.9, 12.2, 10.1, 2.2 Hz, 1H). HRMS: m / z [M + H] + calcd for C 51 H 58 ClN 11 O 11 ,1036.4006; found, 1036.4008.

[0050] 6. Preparation of compound SN38-PABC-K-C6-Tri(a6):

[0051] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-K-C6-N3, to give 315 mg of white solid in 67% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.99 (d, J = 7.4 Hz, 1H), 7.60-7.55 (m, 2H), 7.50 (s, 1H), 7.45-7.33 (m, 4H), 7.14 (d, J = 1.4 Hz, 1H), 6.74 (d, J = 1.0 Hz, 1H), 5.73-5.65 (m, 2H), 5.54 (dq, J = 12.4, 8.1 Hz, 1H), 5.36-5.24 (m, 2H), 5.18 (dd, J = 18.3, 1.0 Hz, 1H), 5.10 (s, 1H), 4.96 (dd, J = 18.3, 1.0 Hz, 1H), 4.86-4.79 (m, 1H), 4.64-4.41 (m, 4H), 3.67-3.56 (m, 1H), 3.31 (t, J = 12.4 Hz, 1H), 2.99-2.75 (m, 3H), 2.61-2.37 (m, 4H), 2.21 (ddd, J = 10.7, 7.2, 5.1 Hz, 1H), 2.10 (tt, J = 12.3, 2.7 Hz, 1H), 1.94-1.77 (m, 3H), 1.79-1.70 (m, 2H), 1.62-1.45 (m, 3H), 1.48-1.25 (m, 9H), 1.14 (s, 2H), 1.00 (t, J = 8.0 Hz, 3H). HRMS: m / z [M+H] + calcd for C 51 H 58 ClN 11 O 11 ,1036.4006; found, 1036.4208.

[0052] 7. Preparation of compound CPT-PABC-AV-Ben-Tri(a7):

[0053] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting materials were CPT-PABC-AV-Ben-N3 and 2-chloro-4-(prop-2-yn-1-yloxy)-1,3,5-triazine, to give 187 mg of white solid in 80% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.96 - 9.75 (m, 1H), 8.98 (d, J = 9.8 Hz, 2H), 8.64 (s, 1H), 8.51 (dd, J = 35.9, 8.1 Hz, 1H), 8.10 (dt, J = 17.3, 8.6 Hz, 4H), 8.00 (d, J = 8.2 Hz, 2H), 7.88 - 7.79 (m, 1H), 7.72 - 7.59 (m, 2H), 7.53 (t, J = 8.2 Hz, 1H), 7.27 (d, J = 6.9 Hz, 2H), 7.01 (s, 1H), 5.68 (t, J = 1.8 Hz, 2H), 5.50 (s, 2H), 5.25 (d, J = 5.4 Hz, 2H), 5.13 - 5.05 (m, 2H), 4.45 - 4.32 (m, 2H), 2.19 - 2.06 (m, 3H), 1.28 (d, J = 7.4 Hz, 3H), 0.90 (dd, J = 23.3, 8.1 Hz, 9H). MS (ESI) m / z: 982.29 [M+H] + .

[0054] 8. Preparation of compound SN38-PABC-AV-C6-Tri(a8)

[0055] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-AV-C6-N3, to give 247 mg of white solid in 53% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.03 (s, 1H), 7.78 (d, J = 7.4 Hz, 1H), 7.52-7.43 (m, 4H), 7.43-7.38 (m, 2H), 7.23 (d, J = 1.5 Hz, 1H), 7.01 (dd, J = 7.5, 1.5 Hz, 1H), 6.60 (dt, J = 12.4, 0.9 Hz, 1H), 5.43 (t, J = 1.1 Hz, 1H), 5.07-4.94 (m, 3H), 4.80-4.66 (m, 2H), 4.66-4.50 (m, 4H), 4.40 (d, J = 9.5 Hz, 1H), 4.37-4.23 (m, 2H), 4.19 (dt, J = 12.4, 3.1 Hz, 1H), 3.44 (dq, J = 12.4, 8.0 Hz, 1H), 3.33 (d, J = 1.0 Hz, 2H), 3.21 (ddd, J = 12.6, 11.4, 2.1 Hz, 1H), 2.93-2.82 (m, 2H), 2.67-2.55 (m, 2H), 2.55-2.43 (m, 2H), 2.20 (dq, J = 12.5, 8.0 Hz, 1H), 2.03-1.87 (m, 3H), 1.83-1.71 (m, 1H), 1.66 (ddt, J = 12.2, 8.5, 2.5 Hz, 1H), 1.39-1.32 (m, 4H), 1.35-1.25 (m, 6H), 1.10-0.90 (m, 10H). HRMS: m / z [M + H] + calcd for C 53 H 62 ClN 11 O 11 ,1064.4319; found, 1064.4320.

[0056] 9. Preparation of compound MMAE-PABC-AV-C6-Tri(a9):

[0057] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to MMAE-PABC-AV-C6-N3, to give 226 mg of white solid in 58% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.28 (s, 1H), 7.83-7.75 (m, 4H), 7.64 (dt, J = 7.6, 1.2 Hz, 2H), 7.50 (d, J = 3.2 Hz, 3H), 7.40-7.27 (m, 3H), 6.77 (s, 1H), 5.43 (q, J = 6.8 Hz, 1H), 5.05 (d, J = 7.0 Hz, 1H), 4.99-4.80 (m, 3H), 4.76 (ddd, J = 12.5, 3.8, 1.8 Hz, 1H), 4.59-4.46 (m, 4H), 4.36-4.22 (m, 2H), 4.16-3.99 (m, 3H), 3.89 (q, J = 7.0 Hz, 1H), 3.49-3.41 (m, 1H), 3.40 (s, 3H), 3.33-3.24 (m, 1H), 3.15 (s, 3H), 2.97 (s, 3H), 2.87-2.73 (m, 2H), 2.73-2.62 (m, 2H), 2.65-2.50 (m, 5H), 2.49 (s, 3H), 2.24-2.07 (m, 2H), 2.02 (hd, J = 6.8, 0.9 Hz, 1H), 1.94-1.64 (m, 7H), 1.36 (d, J = 6.9 Hz, 3H), 1.29 (t, J = 8.0 Hz, 3H), 1.26-1.06 (m, 2H), 1.01 (dd, J = 15.3, 6.8 Hz, 6H), 0.87 (dd, J = 6.8, 4.7 Hz, 9H), 0.82-0.68 (m, 9H), 0.56 (d, J = 6.8 Hz, 3H), 0.47 (t, J = 8.0 Hz, 3H). MS (ESI): m / z: 1403.74 [M+H] + .

[0058] 10. Preparation of compound FL118-PABC-AV-Ben-Tri (a10):

[0059] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting materials were FL118-PABC-AV-Ben-N3 and 2-chloro-4-(prop-2-yn-1-yloxy)-1,3,5-triazine, to give 3156 mg of white solid in 72% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.98 - 9.74 (m, 1H), 9.07 - 8.94 (m, 2H), 8.52 - 8.40 (m, 2H), 8.10 (dt, J = 7.2, 3.7 Hz, 2H), 8.03 (dd, J = 8.6, 2.9 Hz, 2H), 7.58 - 7.44 (m, 4H), 7.28 (q, J = 8.4 Hz, 2H), 6.92 (d, J = 12.0 Hz, 1H), 6.26 (dd, J = 7.1, 2.8 Hz, 2H), 5.67 (t, J = 1.9 Hz, 2H), 5.48 (s, 2H), 5.20 (d, J = 5.3 Hz, 2H), 5.13 - 5.04 (m, 2H), 4.44 - 4.33 (m, 2H), 2.18 - 2.09 (m, 3H), 1.30 (d, J = 7.1 Hz, 3H), 0.99 - 0.87 (m, 9H). MS (ESI): m / z: 1026.28 [M+H] + .

[0060] 11. Preparation of compound CPT-PABC-AV-4PEG-Tri(a11):

[0061] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting materials were CPT-PABC-AV-4PEG-N3 and 2-chloro-4-(prop-2-yn-l-yloxy)-l,3,5-triazine, to give 423 mg of white solid in 75% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.88 (dd, J = 77.5, 8.1 Hz, 1H), 8.88 (dd, J = 7.7, 3.5 Hz, 1H), 8.64 (d, J = 7.8 Hz, 2H), 8.51 - 8.40 (m, 1H), 8.35 - 8.29 (m, 1H), 8.19 - 8.06 (m, 3H), 8.02 (dt, J = 8.7, 3.4 Hz, 2H), 7.89 - 7.82 (m, 1H), 7.71 - 7.61 (m, 2H), 7.56 (dd, J = 11.0, 8.2 Hz, 1H), 7.30 (td, J = 9.7, 5.5 Hz, 2H), 7.04 (d, J = 3.5 Hz, 1H), 5.51 (s, 2H), 5.26 (s, 2H), 5.10 (ddt, J = 17.8, 8.0, 5.1 Hz, 2H), 4.62 (d, J = 2.5 Hz, 2H), 4.48 - 4.36 (m, 2H), 3.61 (dt, J = 5.8, 3.3 Hz, 2H), 3.53 (dt, J = 19.5, 3.3 Hz, 8H), 3.48 - 3.39 (m, 3H), 2.16 (p, J = 6.9 Hz, 3H), 1.31 (d, J = 5.9 Hz, 3H), 1.01 - 0.88 (m, 9H). MS (ESI): m / z: 1144.34 [M+H] + .

[0062] 12. Preparation of compound SN38-PABC-AV-3PEG-Tri(a12)

[0063] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-AV-3PEG-N3, to give 256 mg of white solid, yield 53%. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.37 - 7.22 (m, 6H), 7.20 (s, 1H), 6.68 - 6.61 (m, 1H), 5.78 (ddd, J = 9.5, 2.3, 1.1 Hz, 1H), 5.64 (ddd, J = 9.5, 2.0, 1.1 Hz, 1H), 5.25 - 5.13 (m, 4H), 5.15 - 5.07 (m, 2H), 4.96 (dd, J = 18.6, 1.0 Hz, 1H), 4.76 (ddd, J = 12.2, 11.2, 4.7 Hz, 1H), 4.65 - 4.49 (m, 2H), 4.31 - 4.10 (m, 3H), 3.93 (ddd, J = 12.5, 11.2, 2.8 Hz, 1H), 3.78 - 3.64 (m, 2H), 3.59 - 3.32 (m, 5H), 3.32 - 3.15 (m, 4H), 3.03 - 2.88 (m, 2H), 2.52 (dq, J = 12.4, 8.1 Hz, 1H), 2.16 - 2.02 (m, J = 6.9 Hz, 1H), 1.90 (dq, J = 12.4, 8.0 Hz, 1H), 1.43 (t, J = 8.0 Hz, 3H), 1.36 (d, J = 6.9 Hz, 3H), 1.29 (t, J = 8.0 Hz, 3H), 1.01 (dd, J = 15.0, 6.8 Hz, 6H), 0.80 (t, J = 8.0 Hz, 3H). MS (ESI): m / z: 1096.42 [M+H]+.

[0064] 13. Preparation of compound SN38-PABC-AV-4PEG-Tri(a13)

[0065] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-AV-4PEG-N3, to give 315 mg of white solid, 43% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.03 (d, J = 7.4 Hz, 1H), 7.69-7.64 (m, 2H), 7.55-7.45 (m, 3H), 7.37 (dd, J = 7.5, 1.5 Hz, 1H), 7.14 (d, J = 1.6 Hz, 1H), 6.64 (s, 1H), 6.50 (dt, J = 12.6, 1.2 Hz, 1H), 5.14 (dd, J = 15.9, 1.0 Hz, 1H), 5.09 (s, 1H), 5.07-4.94 (m, 2H), 4.84-4.69 (m, 3H), 4.69-4.47 (m, 3H), 4.42 (ddd, J = 12.5, 11.0, 3.1 Hz, 1H), 4.15 (dq, J = 12.4, 8.0 Hz, 1H), 3.80-3.45 (m, 6H), 3.36-3.21 (m, 4H), 3.25 (s, 3H), 3.16 (d, J = 6.9 Hz, 1H), 3.02-2.87 (m, 2H), 2.87-2.70 (m, 2H), 2.62 (dq, J = 12.2, 8.0 Hz, 1H), 1.91-1.77 (m, J = 6.8 Hz, 1H), 1.76 (s, 1H), 1.39-1.25 (m, 9H), 1.03-0.93 (m, 6H), 0.70 (d, J = 6.8 Hz, 3H). MS (ESI): m / z: 1140.42 [M+H]+.

[0066] 14. Preparation of compound SN38-PABC-Cit-V-C6-Tri (a14)

[0067] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-Cit-V-C6-N3, to give 264 mg of white solid, yield 52%. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.35 (s, 1H), 8.15 (d, J = 1.5 Hz, 1H), 8.06 (d, J = 7.5 Hz, 1H), 7.64-7.59 (m, 2H), 7.52 (d, J = 2.4 Hz, 1H), 7.50 (s, 2H), 7.42-7.31 (m, 3H), 7.06 (t, J = 1.5 Hz, 1H), 5.83 (dd, J = 12.3, 1.1 Hz, 1H), 5.58 (t, J = 7.0 Hz, 1H), 5.53 (s, 2H), 5.23-5.09 (m, 2H), 5.15 (s, 1H), 5.03-4.94 (m, 3H), 4.79 (ddd, J = 12.2, 11.2, 4.8 Hz, 1H), 4.64 (dq, J = 12.5, 8.0 Hz, 1H), 4.58-4.46 (m, 3H), 4.41-4.28 (m, 2H), 3.24 (dd, J = 12.3, 10.1 Hz, 1H), 2.98 (tdd, J = 11.9, 4.9, 2.3 Hz, 1H), 2.90-2.70 (m, 3H), 2.69-2.39 (m, 3H), 2.21-2.04 (m, 3H), 2.01-1.78 (m, 4H), 1.72-1.39 (m, 5H), 1.29 (t, J = 8.0 Hz, 3H), 0.99 (d, J = 6.9 Hz, 3H), 0.96-0.87 (m, 6H), 0.84-0.72 (m, 4H). MS (ESI): m / z: 1164.45 [M+H] + .

[0068] 15. Preparation of compound SN38-PABC-Cit-V-3PEG-Tri(a15):

[0069] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-Cit-V-3PEG-N3, to give 295 mg of white solid in 51% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.42 (s, 2H), 7.99 (d, J = 7.5 Hz, 1H), 7.60-7.54 (m, 2H), 7.52-7.46 (m, 3H), 7.46-7.40 (m, 2H), 7.32 (dd, J = 7.5, 1.5 Hz, 1H), 7.18 (d, J = 1.5 Hz, 1H), 5.67 (d, J = 9.4 Hz, 1H), 5.57 (s, 1H), 5.36 (dt, J = 12.4, 1.1 Hz, 1H), 5.26 (d, J = 12.3 Hz, 1H), 5.22-5.14 (m, 2H), 5.11 (s, 1H), 5.06 (s, 1H), 5.03-4.92 (m, 2H), 4.87 (td, J = 12.3, 2.9 Hz, 1H), 4.79 (d, J = 6.9 Hz, 1H), 4.67-4.58 (m, 2H), 4.51 (qd, J = 7.9, 0.9 Hz, 2H), 4.42-4.26 (m, 3H), 3.96-3.84 (m, 2H), 3.79 (dt, J = 12.3, 1.7 Hz, 1H), 3.65-3.54 (m, 1H), 3.51-3.25 (m, 6H), 3.05 (ddd, J = 12.2, 10.5, 1.8 Hz, 1H), 2.99-2.87 (m, 3H), 2.74-2.55 (m, 3H), 2.13-2.03 (m, 1H), 1.96 (q, J = 8.0 Hz, 2H), 1.87 (td, J = 5.4, 3.3 Hz, 2H), 1.73-1.60 (m, 1H), 1.31 (dt, J = 22.3, 8.1 Hz, 6H), 1.00 (t, J = 8.0 Hz, 3H), 0.89 (dd, J = 25.5, 6.8 Hz, 6H). MS (ESI): m / z: 1297.42 [M+H]+.

[0070] 16. Preparation of compound SN38-PABC-Cit-V-4PEG-Tri (a16):

[0071] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-Cit-V-4PEG-N3, to give 342 mg of white solid in 42% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.06 (d, J = 7.5 Hz, 1H), 7.70-7.65 (m, 2H), 7.50 (d, J = 1.8 Hz, 3H), 7.42-7.34 (m, 3H), 5.89 (dt, J = 12.5, 1.1 Hz, 1H), 5.81 (s, 1H), 5.73 (dt, J = 9.4, 1.7 Hz, 1H), 5.62 (dt, J = 9.5, 1.7 Hz, 1H), 5.46-5.37 (m, 3H), 5.36-5.23 (m, 2H), 5.26 (s, 1H), 5.10 (s, 1H), 4.95 (dd, J = 15.3, 1.0 Hz, 1H), 4.82-4.57 (m, 5H), 4.43 (d, J = 12.5 Hz, 1H), 4.38-4.17 (m, 5H), 3.82-3.70 (m, 2H), 3.72-3.59 (m, 2H), 3.62-3.54 (m, 2H), 3.55-3.44 (m, 4H), 3.44-3.33 (m, 2H), 3.24 (dddd, J = 14.1, 10.8, 3.4, 1.5 Hz, 2H), 3.22-3.07 (m, 4H), 2.99-2.85 (m, 2H), 2.77-2.57 (m, 2H), 2.51 (ddd, J = 12.3, 3.9, 0.9 Hz, 1H), 1.86 (tt, J = 6.4, 5.2 Hz, 2H), 1.53-1.35 (m, 4H), 1.29 (t, J = 8.0 Hz, 3H), 1.06-0.94 (m, 6H), 0.89 (d, J = 6.7 Hz, 3H). MS (ESI): m / z: 1341.52 [M+H]+.

[0072] 17. Preparation of compound SN38-PABC-GFGG-C6-Tri(a17)

[0073] Prepared in the same manner as the procedure of Example 1 for 1, except that the starting material 10-SN38-PABC-C6-N3 was changed to SN38-PABC-GFGG-C6-N3, to give 327 mg of white solid in 67% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.27 (s, 1H), 8.10 (d, J = 1.5 Hz, 1H), 8.03 (d, J = 7.5 Hz, 1H), 7.98 (s, 1H), 7.54 - 7.46 (m, 4H), 7.27 (dd, J = 7.4, 1.6 Hz, 1H), 7.19 (s, 6H), 7.18 - 7.13 (m, 2H), 6.74 (d, J = 5.0 Hz, 3H), 5.50 (t, J = 6.9 Hz, 1H), 5.19 (t, J = 7.0 Hz, 1H), 5.09 (dd, J = 18.2, 1.0 Hz, 1H), 5.05 - 4.88 (m, 4H), 4.61 (dq, J = 12.4, 8.0 Hz, 1H), 4.46 - 4.36 (m, 2H), 4.38 - 4.22 (m, 5H), 4.00 (d, J = 12.4 Hz, 1H), 3.09 (dd, J = 12.4, 7.0 Hz, 1H), 3.05 - 2.94 (m, 2H), 2.68 (tdd, J = 12.3, 4.1, 2.0 Hz, 1H), 2.56 - 2.45 (m, 2H), 2.49 - 2.34 (m, 2H), 2.37 - 2.19 (m, 2H), 1.98 - 1.64 (m, 4H), 1.47 - 1.36 (m, 2H), 1.29 (t, J = 8.0 Hz, 3H), 1.18 (t, J = 8.0 Hz, 3H), 0.97 (t, J = 8.0 Hz, 3H). MS (ESI): m / z: 1226.43 [M+H] + .

[0074] 18. Preparation of 10,11-methylenedioxy-7-(3-furyl)camptothecin-PABC-K-8PEG-Ma (a18):

[0075] 10,11-methylenedioxy-7-(3-furyl)camptothecin-PABC-K-8PEG-N3 (528 mg, 0.43 mmol) was placed in a 10 mL single-necked flask, dissolved in 2 mL of DMSO (Dimethyl sulfoxide), weighed 4-((2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)methyl)-N-(prop-2-yn-1-yl)cyclohexanecarboxamide (354 mg, 1.29 mmol), and cuprous bromide (185 mg, 1.29 mmol) was added to the above system, and the above system was reacted at room temperature for 30 min. The reaction solution was extracted with DCM three times, the organic phases were combined, and the solvent was evaporated under reduced pressure. The product was obtained by preparative liquid chromatography, and white solid 258 mg was obtained after freeze-drying, with a yield of 40%. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.72-7.66 (m, 2H), 7.66-7.57 (m, 2H), 7.56 (td, J = 6.3, 1.4 Hz, 4H), 7.50 (s, 2H), 7.15-7.09 (m, 2H), 6.96 (d, J = 10.8 Hz, 1H), 6.34 (s, 1H), 6.02-5.90 (m, 5H), 5.05 (dd, J = 2.6, 0.9 Hz, 2H), 4.87 (t, J = 5.8 Hz, 3H), 4.75 (d, J = 12.5 Hz, 1H), 4.62-4.48 (m, 4H), 4.46 (s, 2H), 4.05 (d, J = 12.5 Hz, 1H), 3.97-3.87 (m, 3H), 3.67 (ddd, J = 12.5, 11.3, 3.4 Hz, 1H), 3.61 (s, 12H), 3.55 (ddd, J = 12.5, 11.4, 3.8 Hz, 1H), 3.46-3.38 (m, 4H), 3.26 (s, 4H), 3.29-3.10 (m, 5H), 2.65-2.50 (m, 2H), 2.50-2.34 (m, 2H), 2.21-2.05 (m, 2H), 2.00-1.89 (m, 4H), 1.87-1.66 (m, 2H), 1.70-1.48 (m, 2H), 1.41-1.17 (m, 4H), 1.04 (s, 2H), 0.94-0.73 (m, 5H). MS (ESI): m / z: 1502.62 [M+H] + .

[0076] 19. Preparation of compound 10, 11-methylenedioxy-7-(3-thienyl) camptothecin-PABC-AV-C6-Tri(a19)

[0077] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10, 11-methylenedioxy-7-(3-furanyl) camptothecin-PABC-K-8PEG-N3 was changed to 10, 11-methylenedioxy-7-(3-thienyl) camptothecin-PABC-AV-C6-N3, to give 346 mg of white solid in 62% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.21 (s, 1H), 7.72-7.66 (m, 2H), 7.60-7.52 (m, 2H), 7.50 (s, 1H), 7.44-7.34 (m, 3H), 7.27 (d, J = 1.5 Hz, 1H), 7.13 (dd, J = 7.5, 1.5 Hz, 1H), 6.07-5.98 (m, 4H), 5.53 (dt, J = 12.3, 0.9 Hz, 1H), 5.38 (dt, J = 12.3, 1.0 Hz, 1H), 5.18 (dd, J = 18.1, 1.0 Hz, 1H), 5.06-4.84 (m, 4H), 4.77 (d, J = 7.0 Hz, 1H), 4.67 (dq, J = 12.4, 8.0 Hz, 1H), 4.43-4.24 (m, 3H), 3.05 (td, J = 12.1, 3.0 Hz, 1H), 2.75 (tdd, J = 12.1, 3.6, 1.4 Hz, 1H), 2.67-2.56 (m, 2H), 2.56-2.35 (m, 3H), 2.14 (dtdd, J = 13.1, 6.1, 4.3, 2.0 Hz, 1H), 1.80-1.63 (m, 3H), 1.36 (d, J = 6.9 Hz, 3H), 1.29 (t, J = 8.0 Hz, 3H), 1.20-1.04 (m, 4H), 0.99 (t, J = 8.0 Hz, 3H), 0.78 (d, J = 6.9 Hz, 3H). MS (ESI): m / z: 1160.45 [M+H] + .

[0078] 20. Preparation of compound 10, 11-methylenedioxy-7-(1-(3-methylbenzyl)-1H-1,2,3- triazol-4-yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-Tri(a20):

[0079] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10, 11-methylenedioxy-7-(3-furyl)camptothecin-PABC-K-8PEG-N3 was changed to 10, 11-methylenedioxy-7-(1-(3-methylbenzyl)-1H-1,2,3-triazol-4- yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-N3, to give 374 mg of white solid in 57% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 8.21 (s, 1H), 7.75-7.66 (m, 3H), 7.58 (s, 1H), 7.54-7.47 (m, 4H), 7.30-7.20 (m, 2H), 7.18-7.13 (m, 1H), 7.11 (s, 1H), 7.02 (dtd, J = 7.3, 2.2, 1.2 Hz, 1H), 6.29-6.22 (m, 2H), 6.22-6.15 (m, 1H), 6.08-6.00 (m, 2H), 5.93 (dt, J = 2.3, 1.0 Hz, 2H), 5.74 (d, J = 9.4 Hz, 1H), 5.33-5.26 (m, 1H), 5.14 (dd, J = 17.5, 1.0 Hz, 1H), 5.05-4.94 (m, 3H), 4.86 (td, J = 12.1, 3.7 Hz, 1H), 4.77 (d, J = 6.9 Hz, 1H), 4.55-4.37 (m, 4H), 4.27 (d, J = 12.3 Hz, 1H), 4.23-4.11 (m, 2H), 3.99 (dq, J = 12.4, 8.0 Hz, 1H), 3.04 (td, J = 12.1, 3.0 Hz, 1H), 2.76-2.51 (m, 4H), 2.47-2.39 (m, 1H), 2.36 (s, 2H), 2.23-2.11 (m, 1H), 2.09 (s, 1H), 1.93-1.81 (m, 1H), 1.73-1.64 (m, 2H), 1.36 (d, J = 6.9 Hz, 3H), 1.29 (t, J = 8.0 Hz, 3H), 1.09-0.96 (m, 4H), 0.85-0.75 (m, 6H). MS (ESI): m / z: 1292.67 [M+H] + .

[0080] 21. Preparation of compound 10, 11-methylenedioxy-7-(1-(3-fluorobenzyl)-1H-1,2,3- triazol-4-yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-Tri (a21):

[0081] Prepared in the same manner as the procedure for 1 in Example 1, except that the starting material 10, 11-methylenedioxy-7-(3-furanyl)camptothecin-PABC-K-8PEG-N3 was changed to 10, 11-methylenedioxy-7-(1-(3-fluorobenzyl)-1H-1,2,3-triazol-4- yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-N3 to give 385 mg of white solid in 54% yield. 1H NMR (400 MHz, DMSO-d6) δ 9.82 (s, 1H), 9.50 (s, 1H), 7.76 (d, J = 1.1 Hz, 1H), 7.66 - 7.60 (m, 3H), 7.58 - 7.52 (m, 3H), 7.50 (s, 1H), 7.31 (td, J = 7.6, 5.8 Hz, 1H), 7.14 - 6.98 (m, 3H), 6.34 (dt, J = 12.5, 1.2 Hz, 1H), 6.02 (dd, J = 9.5, 1.2 Hz, 1H), 6.00 (s, 2H), 5.92 - 5.78 (m, 3H), 5.59 (dq, J = 12.3, 7.9 Hz, 1H), 5.47 (d, J = 12.4 Hz, 1H), 5.20 (ddd, J = 12.4, 5.9, 1.0 Hz, 1H), 5.09 - 4.94 (m, 4H), 4.81 - 4.63 (m, 3H), 4.55 (dq, J = 12.4, 8.0 Hz, 1H), 4.44 - 4.32 (m, 2H), 4.10 (ddd, J = 12.5, 4.7, 1.5 Hz, 1H), 4.01 (dd, J = 12.4, 1.0 Hz, 1H), 3.20 - 3.12 (m, 1H), 2.67 - 2.51 (m, 2H), 2.17 (td, J = 12.3, 4.5 Hz, 1H), 2.06 (s, 1H), 1.95 (t, J = 8.0 Hz, 2H), 1.76 - 1.58 (m, 2H), 1.42 - 1.17 (m, 10H), 0.99 (d, J = 6.7 Hz, 3H), 0.92 - 0.79 (m, 6H). MS (ESI): m / z: 1296.47 [M+H] + .

[0082] Synthesis of homing peptide (P) of Example 2:

[0083] The template peptide and active peptides (c(RGDfC), SSTR, SSTR-1, CHN-1, 2YHN-1 or 2CHN-1) used in this example were all manually synthesized by standard Fmoc solid phase polypeptide synthesis (Fmoc-SPPS) method.

[0084] Preparation of polypeptide-cell toxin conjugate (PDC) of Example 3

[0085] 1. Preparation of FL118-S-S-c(RGDfC) (b1):

[0086] Take 4 mg c(RGDfC) (1 eq) dissolved in 500 μL pH = 8 PBS (Phosphate buffered saline) solution, add 6.8 mg FL118-S-S-Tri(a4, 1 eq) and pyridine or N, N-dimethylpyridine (2 eq) mixed, the system becomes clear, continue to shake the reaction 2 h, dilute with water and acetonitrile, and then prepare liquid phase. Mobile phase A: 0.05% TFA pure water, mobile phase B: 0.05% TFA acetonitrile. Absorbance: 254 nm and 365 nm. Preparation of mobile phase gradient method: flow rate 6 mL / min, 0-20 min, A = 90%-60%; 20-50 min, A = 60%-40%. The collection solution was freeze-dried in a freeze dryer at -80°C, and 6.3 mg of light yellow solid was obtained with a yield of 60%, and the HPLC (High Performance Liquid Chromatography) purity was identified as 97.2%. HRMS m / z: [M+H] + calcd for C 54 H 61 O 15 N 14 S3,1241.3597; found 1241.3629. purity 97.2%.

[0087] 2, Preparation of compound CPT-PABC-AV-Ben-c(RGDfC) (b2):

[0088] Prepared in the same way as the procedure of 1 in Example 2, except that the raw material a4 was changed to a7, and 13 mg of white solid was obtained with a yield of 47%. HRMS m / z: [M+H] + calcd for C 73 H 78 O 17 N 19 S,1524.5698; found 1524.5720. purity 98.1%.

[0089] 3, Preparation of compound FL118-PABC-AV-Ben-c(RGDfC) (b3):

[0090] Prepared in the same way as the procedure of 1 in Example 2, except that the raw material a4 was changed to a10, and 5.4 mg of light yellow solid was obtained with a yield of 50%. HRMS m / z: [M+H] + calcd for C 73 H 78 O 19 N 19S, 1567.5596; found 1567.5637. purity 98.9%.

[0091] 4. Preparation of compound CPT-PABC-AV-4PEG-c(RGDfC) (b4):

[0092] Prepared in the same manner as for 1 in Example 2, except that the starting material a4 was changed to a11, to give a light yellow solid 11.5 mg, 63% yield. HRMS m / z: [M+H] + calcd for C 79 H 91 O 19 N 20 S, 1655.6485; found 1655.6514. purity 97.9%.

[0093] 5. Preparation of compound SN38-PABC-K-C6-CHN-1 (b5):

[0094] Prepared in the same manner as for 1 in Example 2, except that the starting material was a6 and CHN-1, to give a white solid 8.5 mg, 43% yield. HRMS m / z: [M+H] + calcd for C 111 H 159 N 31 O 29 S, 2424.1674; found 2424.1773. purity 98.9%.

[0095] 6. Preparation of compound SN38-PABC-AV-C6-CHN-1 (b6):

[0096] Prepared in the same manner as for 1 in Example 2, except that the starting material was a8 and CHN-1, to give a white solid 6.3 mg, 53% yield. HRMS m / z: [M+H] + calcd for C 113 H 163 N 31 O 29 S, 2452.1987; found 2452.2087. purity 98.6%.

[0097] 7. Preparation of compound MMAE-PABC-AV-C6-CHN-1 (b7):

[0098] Prepared in the same manner as in Example 2, Step 1, except starting with a9 and CHN-1, to give 5.7 mg of white solid in 62% yield. HRMS m / z: [M+H] + calcd for C 130 H 208 N 34 O 32 S, 2791.5448; found 2791.5737. purity 97.3%.

[0099] 8. Compound SN38-PABC-AV-3PEG-CHN-1 (b8):

[0100] Prepared in the same manner as in Example 2, Step 1, except starting with a12 and CHN-1, to give 4.2 mg of white solid in 42% yield. HRMS m / z: [M+H] + calcd for C 117 H 168 N 32 O 34 S, 2599.2155; found 2599.2348. purity 97.9%.

[0101] 9. Preparation of compound SN38-PABC-AV-4PEG-CHN-1 (b9):

[0102] Prepared in the same manner as in Example 2, Step 1, except starting with a13 and CHN-1, to give 2.2 mg of white solid in 36% yield. HRMS m / z: [M+H] + calcd for C 119 H 172 N 32 O 35 S, 2643.2417; found 2643.2754. purity 98.5%.

[0103] 10. Preparation of compound SN38-PABC-Cit-V-C6-CHN-1 (b10):

[0104] Prepared in the same manner as in Example 2, Step 1, except starting with a14 and CHN-1, to give 3.5 mg of white solid in 32% yield. HRMS m / z: [M+H] + calcd for C 116 H 167 N 33 O 31S, 2252.2260; found 2252.2347. purity 96.4%.

[0105] 11. Preparation of compound SN38-PABC-Cit-V-3PEG-CHN-1 (bl l):

[0106] Prepared in the same manner as for 1 in Example 2, except that the starting material was a 15 and CHN-1, to give a white solid 3.3 mg, 32% yield. HRMS m / z: [M+H] + calcd for C 120 H 174 N 34 O 35 S, 2685.2385; found 2685.2739. purity 97.4%.

[0107] 12. Preparation of compound SN38-PABC-Cit-V-4PEG-CHN-1 (bl2):

[0108] Prepared in the same manner as for 1 in Example 2, except that the starting material was a 16 and CHN-1, to give a white solid 2.5 mg, 29% yield. HRMS m / z: [M+H] + calcd for C 122 H 178 N 34 O 36 S, 2729.2897; found 2729.2946. purity 97.4%.

[0109] 13. Preparation of compound SN38-PABC-GFGG-C6-CHN-1 (bl3):

[0110] Prepared in the same manner as for 1 in Example 2, except that the starting material was a 17 and CHN-1, to give a white solid 4.1 mg, 42% yield. HRMS m / z: [M+H] + calcd for C 120 H 165 N 33 O 32 S, 2614.2053; found 2614.2147. purity 97.6%.

[0111] 14. Preparation of compound SN38-PABC-AV-C6-2YHN-1 (bl4):

[0112] Prepared in the same manner as for example 2, step 1, except starting with a8 and 2YHN-1, to give a white solid, 3.7 mg, 37% yield. HRMS m / z: [M+H] + calcd for C 185 H 243 N 45 O 45 , 3816.8143; found 3816.8246. purity 97.9%.

[0113] 15, Preparation of compound SN38-PABC-Cit-V-C6-2YHN-1 (b15):

[0114] Prepared in the same manner as for example 2, step 1, except starting with a14 and 2YHN-1, to give a white solid, 5.2 mg, 41% yield. HRMS m / z: [M+H] + calcd for C 191 H 251 N 49 O 49 , 4017.8722; found 4017.8835. purity 98.2%.

[0115] 16, Preparation of compound SN38-PABC-AV-C6-2CHN-1 (b16):

[0116] Prepared in the same manner as for example 2, step 1, except starting with a8 and 2CHN-1, to give a white solid, 3.2 mg, 36% yield. HRMS m / z: [M+H] + calcd for C 173 H 235 N 45 O 43 S2, 3696.7060; found 3696.7148. purity 97.2%.

[0117] 17, Preparation of compound SN38-PABC-CV-C6-2CHN-1 (b17):

[0118] Prepared in the same manner as for example 2, step 1, except starting with a14 and 2CHN-1, to give a white solid, 2.6 mg, 33% yield. HRMS m / z: [M+H] + calcd for C 179 H 243 N 49 O 47S2, 3896.7606; found 3896.7734. purity 97.8%.

[0119] 18. Preparation of compound SN38-PABC-AV-C6-SSTR (b18):

[0120] Prepared by the same procedure as for 1 in Example 2, except starting with a8 and SSTR, as a white solid, 3.5 mg, 37% yield. HRMS m / z: [M+H] + calcd for C 102 H 125 N 21 O 23 S2, 2077.8732; found 2077.8837. purity 98.4%.

[0121] 19. Preparation of compound SN38-PABC-AV-4PEG-SSTR (b19):

[0122] Prepared by the same procedure as for 1 in Example 2, except starting with a13 and SSTR, as a white solid, 4.2 mg, 42% yield. HRMS m / z: [M+H] + calcd for C 108 H 134 N 22 O 29 S2, 2268.9162; found 2268.9263. purity 98.7%.

[0123] 20. Preparation of compound SN38-PABC-Cit-V-C6-SSTR (b20):

[0124] Prepared by the same procedure as for 1 in Example 2, except starting with a14 and SSTR, as a white solid, 4.8 mg, 44% yield. HRMS m / z: [M+H] + calcd for C 108 H 134 N 22 O 29 S2, 2177.9005; found 2177.9154. purity 99.2%.

[0125] 21. Preparation of compound SN38-PABC-AV-C6-SSTR-1 (b21):

[0126] Prepared by the same procedure as for 1 in Example 2, except that the starting materials were a8 and SSTR-1, to give 3.8 mg of white solid in 41% yield. HRMS m / z: [M+H] + calcd for C 101 H 124 N 22 O 21 S3,2078.8507; found 2078.8625. purity 98.3%.

[0127] 22. Preparation of compound SN38-PABC-Cit-V-C6-SSTR-1 (b22):

[0128] Prepared by the same procedure as for 1 in Example 2, except that the starting materials were a14 and SSTR-1, to give 3.2 mg of white solid in 35% yield. HRMS m / z: [M+H] + calcd for C 122 H 154 N 22 O 33 S3,2178.8780; found 2178.8869. purity 97.4%.

[0129] 23. Preparation of compound SN38-PABC-GFGG-C6-SSTR-1 (b23):

[0130] Prepared by the same procedure as for 1 in Example 2, except that the starting materials were a17 and SSTR-1, to give 4.6 mg of white solid in 42% yield. HRMS m / z: [M+H] + calcd for C 108 H 126 N 24 O 24 S3,2240.8572; found 2240.8674. purity 98.4%.

[0131] 24. Preparation of compound 10,11-methylenedioxy-7-(3-furanyl)camptothecin-PABC-K-8PEG-SSTR-1 (b24):

[0132] SSTR-1 (1 eq) was weighed into 500 μL of pH 8 PBS solution, 10, 11-methylenedioxy-7-(3-furyl) camptothecin-PABC-K-8PEG-Ma (a18, 1 eq) was added and the mixture was shaken until clear. The reaction was continued for 2 h, diluted with water and acetonitrile and injected onto the preparative HPLC. Mobile phase A: 0.05% TFA in water, mobile phase B: 0.05% TFA in acetonitrile. Absorbance: 254 nm and 365 nm. Preparative HPLC gradient method: flow rate 6 mL / min, 0-20 min, A = 90%-60%; 20-50 min, A = 60%-40%. The collected fractions were lyophilized in a freeze dryer at -80 °C. Yellow solid 7.8 mg, yield 65%, HPLC purity 98.5%. HRMS m / z: [M+H] + calcd for C 122 H 154 N 22 O 33 S3,2553.0244; found 2553.0743. purity 98.5%.

[0133] 25. Preparation of compound 10, 11-methylenedioxy-7-(3-thienyl) camptothecin-PABC-AV-C6-SSTR-1 (b25):

[0134] Prepared in the same manner as for 1 in Example 2, except that a18 was replaced by a19, white solid 4.2 mg, yield 41%. HRMS m / z: [M+H] + calcd for C 108 H 126 N 24 O 24 S3,2275.8290; found 2275.8342. purity 99.2%.

[0135] 26. Preparation of compound 10, 11-methylenedioxy-7-(1-(3-methylbenzyl)-1H-1,2,3- triazol-4-yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-SSTR-1 (b26):

[0136] Prepared in the same manner as for 1 in Example 2, except that a18 was replaced by a20, white solid 3.5 mg, yield 35%. HRMS m / z: [M+H] + calcd for C 108 H 126 N 24 O 24S3,2207.9631; found 2207.9742.purity 98.6%.

[0137] 27. Preparation of compound 10,11-methylenedioxy-7-(1-(3-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)amino)methyl)camptothecin-PABC-AV-C6-SSTR-1(b27):

[0138] The preparation was carried out using the same method as step 1 in Example 2, except that the starting material a18 was changed to a21, yielding 3.6 mg of a white solid, with a yield of 32%. HRMS m / z: [M+H] + calcd for C 108 H 126 N 24 O 24 S3,2411.9380; found 2411.9475.purity 99.4%.

[0139] Example 4: In vitro antitumor activity assay of PDCs compounds b1-b27:

[0140] The IC50 of all compounds b1-b27 against selected tumor cells was tested using the CCK-8 assay with an action time of 48 hours. 50 As shown in Table 1, compounds b1-b4 and b18-b27 both exhibited significant activity against A549 cells, with an IC50 concentration of [missing information]. 50 All were <0.160 μM. Compounds b3 and b1, with 10,11-methylenedioxycamptothecin (FL118) as the toxic molecule, showed significantly better activity than compounds b2 and b4, with CPT as the toxic molecule, possibly due to the mechanism of action of the toxic molecules. Meanwhile, compounds b5-b17 exhibited significant biological activity against oral squamous cell carcinoma Cal27 cells, SCC9 cells, and breast cancer 4T1 cells, with IC50 <0.160 μM. 50 All values ​​were <0.330 μM, with compound b15 showing an IC50 effect on Cal27 cells. 50 The IC50 concentration of compound b16 was 0.883 μM, and its effect on Cal27 and SCC9 cells was [not specified]. 50 The IC50 values ​​for 4T1 cells were 0.926 μM and 0.1346 μM, respectively. 50 The value is 0.0728 μM, which has significant research value.

[0141] Table 1. Cytotoxic activity of the compounds against A549, B16F10, Cal27, SCC9, 4T1 tumor cells and normal Hacat and HEK-293 cells. a a Data correspond to mean ± SD (n = 3).

[0142] Example 5: In vivo anti-tumor activity assay of PDC compound b3:

[0143] The PDC compound b3 with better in vitro anti-tumor activity was selected for in vivo anti-tumor activity test. The A549 cells were inoculated subcutaneously in the armpit of nude mice. After 14 days of tumor formation, intraperitoneal administration was started. The groups were blank control, irinotecan (CPT-11 control, 100 mg / kg), 10,11-methylenedioxy camptothecin (FL118 control, 0.75 mg / kg), b3 low dose (3 mg / kg, the molar amount of FL118 contained in b3 was the same as that in the FL118 group), and b3 high dose (6 mg / kg). The results showed that the FL118 control group, b3 low dose group, and b3 high dose group could inhibit tumor growth to varying degrees, with the smallest tumor weight in the b3 high dose group. By calculating the tumor inhibition rate, the b3 low dose group could significantly inhibit tumor growth, with a tumor inhibition rate of 74.5%, which was higher than that of the FL118 control group (66.1%); the tumor inhibition rate of the b3 high dose group was 90%, which was significantly better than that of the other groups (see Figure 1). In terms of tumor volume, the CPT-11 control group was in a state of continuous growth, the b3 low dose group significantly inhibited tumor growth, the b3 high dose group had the smallest tumor volume and showed a trend of regression, which was significantly better than the FL118 control group. In terms of body weight, there was no difference in body weight between the b3 low dose group and the blank control group. Although the body weight of the b3 high dose group was lower than that of the blank control group, it was still higher than that of the FL118 control group and the CPT-11 control group. The body weights of the FL118 control group and the CPT-11 control group were significantly decreased, indicating that they had greater toxicity, while b3 had lower toxicity in mice. The results showed that b3 could target tumor tissue and release FL118 to exert low-toxicity and high-activity anti-tumor effect.

[0144] Example 6: Lesion assay of heart, liver, spleen, lung, and kidney tissues of b3-treated nude mice:

[0145] The heart, liver, spleen, lung, and kidney of each nude mouse treated in Example 5 were qualitatively analyzed by HE staining. The results are shown in Figure 2. The number of immune cells in the white pulp of the spleen in the FL118 control group was significantly increased, indicating that FL118 could cause an inflammatory response. At the same time, lesions appeared in the lung tissue, the alveoli were expanded, the alveolar septum was narrowed and partially ruptured, the expanded alveoli merged into larger cystic cavities, which could cause pulmonary emphysema. The b3 high dose group and the b3 low dose group had no obvious lesions in the organs. This further indicated that the toxicity of b3 in mice was lower than that of the FL118 control group.

[0146] Example 7 Comparison of in vitro plasma stability of compound b3 and c(RGDfC)-MAL-FL118

[0147] The peptide hydrolytic degradation caused by proteases and peptidases and the cleavage of the coupling agent are the main factors leading to the instability of PDCs in vivo. The instability of PDCs in the plasma in vivo is a factor leading to the toxic and side effects of PDC drugs in vivo. Therefore, the evaluation of the stability in the plasma is of great significance. In order to compare the in vitro human plasma stability of PDCs of the coupling agent triazine ring provided in the present embodiment and the maleimide coupling agent (control group), the human plasma stability experiment of b3 and c(RGDfC)-MAL-FL118 (control group) with triazine ring as the linker within 24 h was carried out in the present example, and HPLC was used for analysis and testing (Figure 3). The results show that: b3 is very stable in the first 6 h, only a small part is degraded, the degradation rate is relatively fast from 6 h to 12 h, but only about 20% is degraded, and with the extension of time, there are still 50% of b3 that are not degraded after 24 h, and the half-life t 1 / 2 is about 24 h. While c(RGDfC)-MAL-FL118 is degraded very quickly, about 50% is degraded at about 9 h, the half-life t 1 / 2 is about 9 h, and only about less than 20% is not degraded after 24 h. The experimental results show that the plasma stability of b3 is significantly better than that of c(RGDfC)-MAL-FL118, t 1 / 2 is prolonged from 9 h to 24 h, which is increased by 2.67 times, and compound b3 has reasonable stability for targeted drug delivery.

[0148] The c(RGDfC)-MAL-FL118 is prepared by the same method as the step of 24 in Example 2, and the difference between b3 and c(RGDfC)-MAL-FL118 is that the linker used by b3 is triazine ring, and the linker used by c(RGDfC)-MAL-FL118 is maleimide.

[0149] Example 8 Rat in vivo pharmacokinetic curves of compound b3 and c(RGDfC)-MAL-FL118

[0150] The present embodiment further carries out the b3 and c(RGDfC)-MAL-FL118 rat in-vivo pharmacokinetic study, and gives the drug-time curve. SD rats are intraperitoneally injected with 4.17 mg / kg of b3 and c(RGDfC)-MAL-FL118, and blood is taken from the rat tail vein at 0, 0.083, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 12, 24 h. After the treatment is completed, the content is determined by LC-MS-MS (as shown in FIG. 4). The results show that the blood drug concentration of b3 and c(RGDfC)-MAL-FL118 significantly increases within 0-2 h, and basically reaches the maximum blood drug concentration of 2500 ng / mL at 2 h. The maximum blood drug concentration of b3 can be maintained for 4 h, and the maximum blood drug concentration of c(RGDfC)-MAL-FL118 can be maintained for 2 h. The maximum blood drug concentration of b3 is maintained for twice as long as that of c(RGDfC)-MAL-FL118. The blood drug concentration of c(RGDfC)-MAL-FL118 significantly decreases at 4 h-8 h, and the blood drug concentration of b3 significantly decreases at 6 h-8 h. However, at 6 h, the blood drug concentration of b3 is 1000 ng / mL higher than that of c(RGDfC)-MAL-FL118. After 24 h, the blood drug concentrations of b3 and c(RGDfC)-MAL-FL118 decrease to the vicinity of the minimum detection limit. The in-vivo plasma stability of b3 is better than that of c(RGDfC)-MAL-FL118, which proves that the stability of the triazine coupling agent is better than that of the maleimide coupling agent (control group), and is helpful for the development of PDCs drugs.

Claims

1. A polypeptide conjugate compound, wherein, The structural formula of the compound is shown as formula (I): Wherein, P is a transmembrane peptide or a targeting peptide; D is a load; T-B-Z-L is a linker between P and D; T is Wherein, * connects P, # connects B; X is selected from any one of O, S, NH; R' is selected from any one of the group consisting of -(CH2) n1 -, -(CH2CH2O) n2 -, C3-C8 carbocyclyl, phenyl, and heterocyclyl containing N, S or O, wherein n1 and n2 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11; Y is selected from any one of the group consisting of H, N, S, CH2, and O; R" is selected from any one of the group consisting of alkyl of 1-5 carbon atoms, phenyl, naphthyl, halogen substituted phenyl, styryl, methyl substituted phenyl, alkyl alkyn of 1-10 carbon atoms, and alkyl azide of 1-10 carbon atoms; B is selected from any of the following groups: -(CH2) n3 -、-(CH2CH2O) n4 -、-(CH2) n5 -SS-(CH2) n6 - C3-C8 carbon cycloyl groups and phenyl groups, wherein n3, n4, n5, and n6 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11, respectively; Z is -(AA) n7 - wherein AA is an amino acid or a polypeptide, n7 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20; m is selected from 0 or 1; L is R is selected from any one of the following group: hydrogen, halogen, methyl, ethyl, nitro, methoxy, and ethoxy; q is independently 0, 1, 2, 3 or 4; p is selected from 0 or 1.

2. The polypeptide conjugate compound of claim 1, wherein, P is selected from any one of:

3. The polypeptide conjugate compound of claim 1 or 2, wherein, D is selected from any one of the following group: plinabulin, aplysiatoxin, auristatin, maytansine, MMAE, MMAF, a calicheamicin, a duocarmycin, a pyrrolobenzodiazepine, irinotecan, irinotecan hydrochloride, an exatecan derivative, a camptothecin, 7-ethyl-10-hydroxy camptothecin, 10,11-methylenedioxycamptothecin, 10,11-methylenedioxy-7-(3-furyl)camptothecin, 10,11-methylenedioxy-7-(3-thienyl)camptothecin, 10,11-methylenedioxy-7-(1-(3-methylbenzyl)-1H-1,2,3-triazol-4-yl)methyl)amino)methyl)camptothecin, and 10,11-methylenedioxy-7-(1-(3-fluorobenzyl)-1H-1,2,3-triazol-4-yl)methyl)amino)methyl)camptothecin; 4. The polypeptide conjugate compound of claim 1, wherein, Load D and linker T-B-Z-L to form intermediate T-B-Z-L-D; said intermediate is selected from any one of a1-a21 below:

5. The polypeptide-conjugate compound of claim 1, wherein, The polypeptide conjugate compound is selected from any one of b1-b27:

6. The polypeptide conjugate compound of claim 1, wherein, The AA is selected from any one of the following amino acid or peptide sequence: Val-Cit, Val-Ala, Val-Lys, Phe-Lys, Lys-Lys, Ala-Lys, Phe-Cit, Leu-Cit, Ile-Cit, Trp, Lys, Cit, Phe-Ala, Phe-Phe-Lys, D-Phe-Phe-Lys, Gly-Phe-Lys, Leu-Ala-Leu, Ile-Ala-Leu, Val-Ala-Val, Ala-Leu-Ala-Leu, β-Ala-Leu-Ala-Leu, Gly-Phe-Leu-Gly, and Gly-Gly-Phe-Gly.

7. The polypeptide conjugate compound of claim 1, wherein, The compound at least extends to stereoisomers of the polypeptide conjugate compound, pharmaceutically acceptable salts of the polypeptide conjugate compound, and solvates of the polypeptide conjugate compound.

8. A polypeptide conjugate drug prepared by using the polypeptide conjugate compound of any one of claims 1-7.

9. Use of a polypeptide conjugate compound according to any one of claims 1 to 7, wherein, The polypeptide conjugate compound is used for preparing a drug for preventing or treating a disease or a disorder, or reducing the severity of the disease or disorder.

10. Use according to claim 9, wherein, The disease or disorder is cancer, selected from at least one of the following group: breast cancer, squamous cell carcinoma, lung cancer, peritoneal cancer, liver cancer, gastric cancer, gastrointestinal cancer, membranous adenocarcinoma, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, urethral cancer, hepatoma, intestinal cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer, uterine cancer, salivary gland cancer, kidney cancer, prostate cancer, vulvar cancer, thyroid cancer, anal cancer, penile cancer, melanoma, multiple myeloma, lymphoma, lymphoid tumor, brain cancer, gallbladder cancer, esophageal cancer, bile duct cancer, head and neck cancer, and metastases of various cancers.

11. Use according to claim 10, wherein, The lung cancer is adenocarcinoma of lung, squamous cell carcinoma of lung, small cell lung cancer or non-small cell lung cancer; the breast cancer is HER2-positive breast cancer; the lymphoma is B-cell lymphoma; the squamous cell carcinoma is squamous epithelial cell carcinoma or oral squamous cell carcinoma.

12. The use according to claim 9, wherein, The disease or disorder is an infectious disease selected from at least one of the following group: a respiratory infectious disease, a digestive infectious disease, a contact infectious disease, an insect-borne infectious disease, a blood-borne infectious disease, and a body fluid-borne infectious disease.

Images