CN115990264A - PTK7 targeted aptamer coupled drug - Google Patents
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Abstract
The invention discloses a PTK7 targeted nucleic acid aptamer coupling drug, which belongs to the technical field of biological medicines and comprises a PTK7 targeted nucleic acid aptamer and a cytotoxic drug; the cytotoxic drug is a linker-containing australistatin tubulin inhibitor or derivative thereof. The PTK7 targeting aptamer consists of a sequence of SEQ ID NO.1, and the cytotoxic drug is at least one of auristatin E or a derivative thereof, auristatin F or a derivative thereof and auristatin-0101 or a derivative thereof. The PTK7 targeting aptamer coupling drug is a coupling drug of a target PTK7 aptamer and a cytotoxic drug commonly used in the target drug, namely an Australian-type tubulin inhibitor or a derivative thereof, so that the target delivery of the Australian-type tubulin inhibitor or the derivative thereof is realized, the target treatment of PTK7 high-expression malignant tumor can be realized, and the PTK7 targeting aptamer coupling drug is used for preventing or treating PTK7 high-expression malignant tumor.
Description
Technical Field
The invention belongs to the technical field of biological medicines, and particularly relates to a PTK7 targeted nucleic acid aptamer coupling drug.
Background
A nucleic acid aptamer is an antibody-like oligonucleotide sequence capable of specifically binding to a target protein. The aptamer can be chemically synthesized, is easier to modify and more stable than an antibody, has smaller size and organism immune response, and therefore has wide application in diagnosis and treatment of tumors. The coupling of the drug molecules and the aptamer can improve the water solubility of the drug molecules and the enrichment of the drug molecules in tumor cells, reduce the toxic and side effects of the drug molecules on normal cells, and realize the targeted delivery of the drug molecules.
PTK7, protein tyrosine kinase 7 (Protein tyrosine kinase), is a receptor tyrosine kinase in the Wnt pathway involved in Wnt signaling during hematopoietic and somatic progenitor cells and stem cell development. It is overexpressed in a variety of tumor types, including advanced triple negative breast cancer, non-small cell lung cancer, ovarian cancer, colorectal cancer, gastric cancer, esophageal cancer, and the like. Furthermore, PTK7 expression is associated with lymph node metastasis, while being highly expressed in Tumor Initiating Cells (TICs) or tumor stem cells (CSCs) and stromal cells, which are closely related to tumor recurrence and progression. The development of a targeting drug taking PTK7 as a target is expected to be a difficult problem of limited curative effect of a drug for overcoming malignant tumors such as triple negative breast cancer, non-small cell lung cancer and the like. For example, chinese patent application publication number CN 113491773A discloses an artemisinin derivative aptamer drug conjugate, a preparation method and application thereof, and PTK7 targeted aptamer is coupled with drug molecules combretastatin and artemisinin to construct a PTK7 targeted aptamer conjugate drug, so that effective inhibition of PTK7 high-expression cells and tumors is realized.
Disclosure of Invention
The invention aims to provide a PTK7 targeting aptamer coupling drug which can realize the targeting treatment of PTK7 high-expression malignant tumor and is used for preventing or treating PTK7 high-expression malignant tumor.
The technical scheme adopted by the invention for achieving the purpose is as follows:
a PTK7 targeted nucleic acid aptamer conjugate drug comprising a PTK7 targeted nucleic acid aptamer and a cytotoxic drug comprising a linker; wherein the cytotoxic drug is an auristatin type tubulin inhibitor or a derivative thereof.
The PTK7 targeting aptamer coupling drug is a coupling drug of a target PTK7 aptamer and a cytotoxic drug commonly used in the target drug, namely an Australian-type tubulin inhibitor or a derivative thereof, so that the target delivery of the Australian-type tubulin inhibitor or the derivative thereof is realized, the target treatment of PTK7 high-expression malignant tumor can be realized, and the PTK7 targeting aptamer coupling drug is used for preventing or treating PTK7 high-expression malignant tumor. In addition, the PTK7 targeted nucleic acid aptamer coupled drug is used for targeted treatment of PTK7 high-expression cells and a high-expression tumor model, wherein the PTK7 targeted aptamer coupled drug has a good inhibition effect on triple negative breast cancer SUM149 cells, and the IC50 value is 600nM; the composition has good inhibition effect on the OVCAR3 cells of the ovarian cancer, and the IC50 value is 60nM; the HCT116 cell of colorectal cancer has good inhibition effect, and the IC50 value is 200nM; the PTK7 targeted aptamer-coupled drug can achieve 100% objective response rate and complete remission on a triple-negative breast cancer PDX model, and solves the technical problem that an auristatin microtubulin inhibitor or a derivative thereof has poor drug effect in an animal model.
In a preferred embodiment, the PTK7 targeting aptamer consists of the sequence of SEQ ID No. 1.
In a preferred embodiment, the PTK7 targeting aptamer is a thiol modified PTK7 targeting aptamer. Preferably, the PTK7 targeting aptamer is thiol modified sgc8c, i.e. the PTK7 targeting aptamer is SH-sgc8c.
In a preferred embodiment, the cytotoxic agent is at least one of monomethyl auristatin E or a derivative thereof, monomethyl auristatin F or a derivative thereof, auristatin-0101 or a derivative thereof.
In a preferred embodiment, the cytotoxic drug is VcMMAE.
The invention also provides a preparation method of the PTK7 targeted nucleic acid aptamer coupling drug, which comprises the step of coupling the PTK7 targeted nucleic acid aptamer with an auristatin microtubule inhibitor to obtain the PTK7 targeted nucleic acid aptamer coupling drug.
In a preferred embodiment, the coupling method comprises thiol-maleimide coupling, azide-alkyne coupling or amino-carboxyl coupling.
In a preferred embodiment, the molar ratio of PTK7 targeting aptamer to australistatin tubulin inhibitor is 1:2-6.
In a preferred embodiment, a method of preparing a PTK7 targeted aptamer conjugate drug, comprising,
and mixing the PTK7 targeting aptamer solution and the VcMMAE solution, wherein the molar ratio of the PTK7 targeting aptamer to the VcMMAE is 1:2-6, stirring and reacting for 2-36 hours at the temperature of 4-40 ℃, purifying, and freeze-drying to obtain the PTK7 targeting aptamer coupled drug.
Preferably, the PTK7 targeting aptamer is a sulfhydryl modified sgc8c.
More preferably, the method for synthesizing the sulfhydryl modified Sgc c comprises the following steps:
1) Performing solid phase synthesis by using a DNA synthesizer;
2) Placing 3' thio S-S CPG in a synthesis tank, inputting a corresponding sequence in software to start synthesis, and simultaneously reserving a DMT protecting group on the last base;
3) Taking CPG out, adding 200-500 μl of 20-50wt% concentrated ammonia water, heating at 50-80deg.C for 1-5 hr, adding 20-50 μl of 1-5M NaCl solution and 0.2-2mL absolute ethanol for precipitation, centrifuging, removing supernatant, adding ddH 2 O is redissolved to obtain a DNA crude product;
4) Purification was performed using HPLC and freeze-dried.
5) Adding 60-90wt% acetic acid aqueous solution into the freeze-dried product, treating at 25-50deg.C for 0.5-2 hr, removing DMT protecting group to expose 5-terminal hydroxyl group, and precipitating again to obtain sulfhydryl modified Sgc c.
Preferably, the solvent for the PTK7 targeting aptamer solution is one or a combination of several of water, nuclease-free water, phosphate buffer and triethylammonium acetate buffer. In order to ensure the dissolution of small molecules, more preferably, the aptamer solution contains at least 30v/v% acetonitrile in the solvent.
Preferably, the concentration of PTK7 targeting aptamer solution is 0.1-2mM. More preferably, the concentration of PTK7 targeting nucleic acid aptamer solution is 0.8-1mM.
Preferably, the Vcmmae solution is an aqueous acetonitrile solution. More preferably, the volume ratio of acetonitrile to water acetonitrile/water=1/(0.5-3.0). Further, the volume ratio of acetonitrile to water acetonitrile/water=1/(0.5-1.5).
Preferably, the concentration of VcMMAE solution is 10-50mM. More preferably, the concentration of VcMMAE solution is 35-40mM.
Preferably, the purification is performed by one or more methods selected from high performance liquid chromatography, size exclusion chromatography and gel electrophoresis.
Preferably, high performance liquid chromatography is used for purification, the separation medium is a C18 column, the mobile phase A is TEAA buffer solution, and the mobile phase B is ACN. More preferably, the purification is performed by high performance liquid chromatography with a SHIMADZU C18 column as separation medium, TEAA buffer with pH of 7.0 and 0.1M as mobile phase A, and ACN as mobile phase B.
The invention also provides application of the PTK7 targeted aptamer coupling medicament in preparing medicaments for preventing and/or treating malignant tumors.
In a preferred embodiment, the malignancy is a PTK7 high expressing malignancy.
In a preferred embodiment, the malignancy is breast cancer, ovarian cancer, or colorectal cancer.
Preferably, the breast cancer is caused by triple negative breast cancer SUM149 cells, the ovarian cancer is caused by ovarian cancer OVCAR3 cells, and the colorectal cancer is caused by colorectal cancer HCT116 cells.
The invention also provides application of the PTK7 targeting aptamer in preparing a PTK7 targeting aptamer coupling medicament, wherein the PTK7 targeting aptamer consists of a sequence of SEQ ID NO.1, and the cytotoxic medicament for the PTK7 targeting aptamer coupling medicament is at least one of monomethyl auristatin E or a derivative thereof, monomethyl auristatin F or a derivative thereof and auristatin-0101 or a derivative thereof.
In a preferred embodiment, the PTK7 targeting aptamer is used to enhance targeted delivery of cytotoxic drugs in PTK7 targeting aptamer-coupled drugs.
In a preferred embodiment, the PTK7 targeting aptamer is used to enhance targeted therapy of PTK7 targeting aptamer-coupled drugs.
The invention also provides a medicine for preventing and/or treating malignant tumors, which comprises the PTK7 targeted aptamer coupling medicine.
In a preferred embodiment, the malignancy is a PTK7 high expressing malignancy.
In a preferred embodiment, the malignancy is breast cancer, ovarian cancer, or colorectal cancer.
In a preferred embodiment, the malignancy is a large malignancy with high expression of PTK 7. Preferably, the tumor volume of the large malignant tumor is equal to or more than 400mm 3 . Preferably, the tumor volume of the large malignant tumor is 500-600mm 3 。
The PTK7 targeting aptamer coupling medicament prepared by adopting the PTK7 targeting aptamer and the auristatin microtubulin inhibitor or the derivative thereof has the following beneficial effects: the PTK7 targeted aptamer coupled drug is used for targeted treatment of PTK7 high-expression cells and a high-expression tumor model, wherein the PTK7 targeted aptamer coupled drug has a good inhibition effect on triple negative breast cancer SUM149 cells, and the IC50 value is 600nM; the composition has good inhibition effect on the OVCAR3 cells of the ovarian cancer, and the IC50 value is 60nM; the HCT116 cell of colorectal cancer has good inhibition effect, and the IC50 value is 200nM; the PTK7 targeted aptamer-coupled drug can achieve 100% objective response rate and complete remission on a triple-negative breast cancer PDX model, and solves the technical problem that an auristatin microtubulin inhibitor or a derivative thereof has poor drug effect in an animal model; the PTK7 targeted aptamer-coupled drug has a remarkable inhibition effect on a large tumor living body compared with a standard chemotherapy docetaxel group, achieves an inhibition rate of 82.6%, and effectively inhibits proliferation of PTK7 high-expression cells and high-expression tumors. Therefore, the invention provides the PTK7 targeting aptamer coupling drug which can realize the targeting treatment of the PTK7 high-expression malignant tumor and is used for preventing or treating the PTK7 high-expression malignant tumor.
Drawings
FIG. 1 is a synthetic route to PTK7 targeted aptamer-coupled drugs of the invention;
FIG. 2 is a mass spectrum of a PTK7 targeted aptamer coupled drug of the invention;
FIG. 3 shows the inhibition of SUM149 cell proliferation by the PTK7 targeted aptamer-coupled drug of the invention;
FIG. 4 is a graph showing the inhibition of proliferation of OVCAR3 cells by PTK7 targeted aptamer-coupled drugs of the present invention;
FIG. 5 shows the inhibition of HCT116 cell proliferation by PTK7 targeted aptamer-coupled drugs of the invention;
FIG. 6 is a graph showing the tumor volume of a PTK7 targeted aptamer-coupled drug in vivo anti-tumor experiment of the invention;
FIG. 7 is a graph showing the relative tumor volumes of in vivo anti-tumor experiments with PTK7 targeted aptamer-coupled drugs of the invention;
FIG. 8 is a graph showing the body weight of a PTK7 targeted aptamer-conjugated drug in vivo antitumor test mouse according to the present invention;
FIG. 9 is a graph showing the relative body weight of a PTK7 targeted aptamer-conjugated drug in vivo anti-tumor experimental mouse of the present invention;
FIG. 10 is a graph showing tumor immunohistochemical staining after treatment with the PTK7 targeted nucleic acid aptamer-coupled drug of the invention;
FIG. 11 is a graph showing bone marrow and liver toxicity following treatment with the PTK7 targeted aptamer-coupled drug of the invention;
FIG. 12 is a graph showing the experimental tumor volumes of live anti-large tumors of PTK7 targeted aptamer-coupled drugs of the invention;
FIG. 13 shows the body weight of a PTK7 targeted aptamer-conjugated drug-in-vivo anti-tumor laboratory mouse of the present invention.
Detailed Description
The present invention will be further described in detail with reference to specific embodiments in order to make the objects, technical solutions and advantages of the present invention more apparent.
The experimental methods in the following examples are conventional methods unless otherwise specified. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
Example 1:
a preparation method of a PTK7 targeting aptamer coupling drug is characterized in that a PTK7 targeting aptamer is coupled with an Australian microtubulin inhibitor, the synthetic route is shown in figure 1 (note: a sulfhydryl band is PTK7 targeting aptamer sgc8c, which consists of the sequence of SEQ ID NO.1, specifically 5'-ATC TAA CTG CTG CGC CGC CGG GAA AAT ACT GTA CGG TTA GA-3'), and the synthetic method is as follows:
thiol-modified sgc8c solution (1 equivalent) and VcMMAE solution (3 equivalents) were added to a centrifuge tube and mixed, reacted for 16 hours at 37℃with stirring, then HPLC purified using the conditions of Table 1, and freeze-dried to obtain PTK7 targeted aptamer conjugate drug PTK7-ApDC, 75% yield, desalted and lyophilized for use. Mass spectrum as shown in fig. 2, MS: calculated 14147.1 (Found: 14143.8).
Wherein the solvent for the mercapto-modified sgc8c solution is nuclease-free water, and contains 40v/v% acetonitrile, and the concentration of the mercapto-modified sgc8c solution is 1mM; the VcMMAE solution was an aqueous acetonitrile solution, the volume ratio of acetonitrile to water was acetonitrile/water=1/1, and the concentration of VcMMAE solution was 35mM.
TABLE 1 HPLC purification conditions
Wherein, the synthetic method of the mercapto-modified sgc8c comprises the following steps:
1) Using a DNA synthesizer to carry out solid phase synthesis, and drying and dewatering all reagents and containers;
2) Filling 3' thio S-S CPG, placing in a synthesis tank, inputting a corresponding sequence in software to start synthesis, and simultaneously reserving a DMT protecting group on the last base;
3) CPG was taken out and added with 400. Mu.L of 30% strength ammonia water, heated at 65℃for 2 hours, followed by precipitation with 40. Mu.L of 3M NaCl solution and 1mL of absolute ethanol, centrifugation to remove the supernatant and addition of ddH 2 O is redissolved to obtain a DNA crude product, wherein a DMT group is modified at the 5 'end of the DNA crude product, and a sulfhydryl group is modified at the 3' end of the DNA crude product;
4) Purifying the DNA crude product by HPLC, and freeze-drying to obtain a freeze-dried product; due to the existence of DMT groups, the successfully synthesized product has longer retention time so as to be separated to obtain the product;
5) Adding aqueous solution of acetic acid with concentration of 80% into the freeze-dried product, treating at 37 ℃ for 0.5h, removing DMT protecting group to expose 5' -terminal hydroxyl, and then carrying out precipitation collection again to obtain the sulfhydryl modified Sgc c.
Test example 1:
1. PTK7 targeted aptamer coupling drug and malignant cell binding and proliferation inhibition experiment
Malignant tumor cells to be tested (triple negative breast cancer SUM149 cells, ovarian cancer cells OVCAR3 cells and colorectal cancer HCT116 cells) are respectively inoculated into 96-well plates (8000 cells in each well), cultured for 24 hours at 37 ℃ under the condition of 5% carbon dioxide, and 1640 complete medium (containing 10% FBS) containing PTK7 targeting aptamer conjugate medicines PTK7-ApDC with different concentrations is added. After culturing at 37℃under 5% carbon dioxide for 24 hours, the medicated medium was discarded, and the culture was continued for 72 hours with the medium containing no drug. The inhibitory effect of the drug was measured after 96 hours in total using the MTS kit and the results are shown in FIGS. 3-5.
Fig. 3 shows the result of inhibition of PTK7 targeting aptamer-coupled drug on proliferation of SUM149 cells, and fig. 3 shows that PTK7 targeting aptamer-coupled drug PTK7-ApDC has good inhibition effect on triple negative breast cancer SUM149 cells, and IC50 value of PTK7 targeting aptamer-coupled drug PTK7-ApDC is 600 nM. Fig. 4 shows the inhibition result of PTK7 targeting aptamer-coupled drug on OVCAR3 cell proliferation, and fig. 4 shows that PTK7 targeting aptamer-coupled drug PTK7-ApDC has good inhibition effect on OVCAR3 of ovarian cancer cells, and IC50 value of PTK7 targeting aptamer-coupled drug PTK7-ApDC is 60 nM. Fig. 5 shows the inhibition result of PTK7 targeting aptamer-coupled drug on HCT116 cell proliferation, and fig. 5 shows that PTK7 targeting aptamer-coupled drug PTK7-ApDC has good inhibition effect on colorectal cancer HCT116 cells, and IC50 value of PTK7 targeting aptamer-coupled drug PTK7-ApDC is 200 nM.
2. PTK7 targeted aptamer coupled drug for inhibiting malignant tumor PDX animal model experiment
The mouse model used female NSG mice and triple negative breast cancer PDX tumor mass was transplanted using a trocar. After 15 days, the tumor volume reaches 100-150 mm 3 Then taking a triple negative breast cancer PDX model tumor volume of 100-150 mm 3 Is randomly divided into 2 groups. The two experiments were administered every other day, with saline (saline group) and PTK7 targeted aptamer-conjugated drug PTK7-ApDC (PTK 7-ApDC group) administered at a dose of 0.36 mg/kg equivalent of MMAE once a day for a total of 5 times. The body weight of the mice, the length of the tumor (a) and the width of the tumor (b) were measured and recorded at each injection. The calculation formula of the tumor volume (V) is: v= (a×b) 2 )/2. In a tumor volume of more than 1500 mm 3 Or weight loss of more than 15% was the end point of the experiment, euthanized mice were subjected to termination of the experiment.
The tumor volume and the relative tumor volume of the PTK7 targeting aptamer-coupled drug in vivo antitumor experiment are shown in fig. 6 and 7, respectively, and as can be seen from fig. 6 and 7, the objective response rate of 100% and complete remission of the tumor of the PTK7-ApDC group mice can be realized relative to the sample group.
The body weight and the relative body weight of the PTK7 targeting aptamer-coupled drug in vivo antitumor experiment mice are shown in fig. 8 and 9, respectively, and as can be seen from fig. 8 and 9, the body weight of the PTK7-ApDC group mice is not significantly different from that of the saline group.
The tumor of the PDX model of triple negative breast cancer after PTK7 targeting aptamer-conjugated drug PTK7-ApDC treatment is embedded and subjected to immunohistochemical characterization, and the result is shown in figure 10. As can be seen from fig. 10, the expression of CK19 in tumor tissue after PTK7-ApDC treatment with the PTK7 targeting aptamer-coupled drug was reduced relative to the saline group, indicating that tumor cells after PTK7-ApDC treatment with the PTK7 targeting aptamer-coupled drug were reduced; the reduced Ki67 expression indicates that the proliferation of tumor cells is effectively inhibited after PTK7 targeting aptamer is coupled with a drug PTK7-ApDC treatment; the relative increase in pHH3 indicates that tumor cell mitosis is significantly inhibited following PTK7-ApDC treatment with the PTK7 targeting aptamer-conjugated drug.
The main organs such as liver of the triple negative breast cancer PDX model after PTK7 targeting aptamer coupled with the drug PTK7-ApDC treatment are embedded and then subjected to immunohistochemical characterization, and the result is shown in figure 11. As shown in fig. 11, compared with the saline group, after PTK7 targeting aptamer-coupled drug PTK7-ApDC treatment, sternum marrow was not significantly inhibited, and liver cells were not significantly different from the physiological saline group, indicating that PTK7 targeting aptamer-coupled drug PTK7-ApDC has good safety in mice.
3. PTK7 targeting aptamer coupled drug in vivo treatment experiment
Taking tumor volume of triple negative breast cancer PDX model of 500-600mm 3 Is randomly divided into 2 groups. The two experiments were administered every other day, with the clinical first line chemotherapeutic Docetaxel, PTK7 targeted aptamer-coupled drug PTK7-ApDC, administered at a dose of 20 mg/kg Docetaxel or 0.36 mg/kg equivalent MMAE, administered once a total of 5 times per day, by tail intravenous injection, respectively. Body weight of mice, length of tumor (a) and width of tumor (b) were measured and recorded at each injection. The calculation formula of the tumor volume (V) is: v= (a×b) 2 )/2. In a tumor volume of more than 1500 mm 3 Or weight loss of more than 15% was the end point of the experiment, euthanized mice were subjected to termination of the experiment.
The volume of the experimental tumor of the PTK7 targeting aptamer-coupled drug living anti-large tumor is shown in fig. 12, and as can be seen from fig. 12, compared with the Docetaxel group, the PTK7 targeting aptamer-coupled drug PTK7-ApDC treatment effectively inhibits the large tumor of the mice in the PTK7-ApDC group, and the inhibition rate reaches more than 82.6%. At the same time, the body weight of the mice was observed, and the results are shown in FIG. 13, in which the body weight of the PTK7-ApDC group was not significantly different from that of the Docetxel group.
The reverse phase preparation column purification, desalination, administration of the seed and tail vein, flow cell experiment, cell tumor implantation and the like involved in each step of the method are all conventional methods in the field, and can be determined by a person skilled in the art by combining common knowledge in the field and the description of the invention, and are not repeated herein.
While the foregoing embodiments have been described in detail in connection with the embodiments of the invention, it should be understood that the foregoing embodiments are merely illustrative of the invention and are not intended to limit the invention, and any modifications, additions, substitutions and the like made within the principles of the invention are intended to be included within the scope of the invention.
Claims (10)
1. A PTK7 targeted nucleic acid aptamer conjugate drug comprising a PTK7 targeted nucleic acid aptamer and a cytotoxic drug comprising a linker; the cytotoxic drug is an auristatin type tubulin inhibitor or a derivative thereof.
2. A PTK7 targeted nucleic acid aptamer conjugate drug according to claim 1, wherein: the PTK7 targeting aptamer consists of the sequence of SEQ ID NO. 1.
3. A PTK7 targeted nucleic acid aptamer conjugate drug according to claim 1, wherein: the cytotoxic drug is at least one of monomethyl auristatin E or derivatives thereof, monomethyl auristatin F or derivatives thereof, and auristatin-0101 or derivatives thereof.
4. A method for preparing a PTK7 targeting aptamer coupled drug according to claim 1, 2 or 3, comprising the step of coupling the PTK7 targeting aptamer with an auristatin tubulin inhibitor through a coupling reaction to obtain the PTK7 targeting aptamer coupled drug.
5. The method for preparing a PTK7 targeted aptamer coupled drug according to claim 4, wherein: the coupling reaction comprises mercapto-maleimide coupling, azide-alkyne coupling or amino-carboxyl coupling.
6. The method for preparing a PTK7 targeted aptamer coupled drug according to claim 4, wherein: the molar ratio of the PTK7 targeting aptamer to the auristatin microtubulin inhibitor is 1:2-6.
7. Use of a PTK7 targeted aptamer conjugate drug as claimed in claim 1 or 2 or 3 in the manufacture of a medicament for the prevention and/or treatment of malignant tumors.
8. Use according to claim 7, characterized in that: the malignant tumor is PTK7 high-expression malignant tumor.
9. Use of a PTK7 targeting aptamer in the preparation of a PTK7 targeting aptamer conjugate drug, the PTK7 targeting aptamer consisting of the sequence of SEQ ID No.1, the cytotoxic drug being at least one of monomethyl auristatin E or a derivative thereof, monomethyl auristatin F or a derivative thereof, auristatin-0101 or a derivative thereof.
10. A medicament for preventing and/or treating malignant tumors, comprising the PTK7 targeting aptamer-coupled medicament of claim 1 or 2 or 3.
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