Targeted chemical agents, methods for producing the same, pharmaceutical compositions, and uses of targeted chemical agents.

Abstract

The present invention provides a targeted chemical agent, a method for producing the same, a pharmaceutical composition, and uses for the targeted chemical agent. The targeted chemical agent comprises a targeted nucleic acid carrier and a small molecule compound (small molecule drug) supported on the targeted nucleic acid carrier. The targeted nucleic acid carrier comprises a nucleic acid carrier and a target molecule bound to the nucleic acid carrier, and the nucleic acid carrier comprises a DNA carrier or an RNA carrier or sequences A, B, and C that form a self-assembling structure. The targeted chemical agent can deliver the small molecule compound more stably and reliably, and further exhibits a combination therapeutic effect between the small molecule compound and other drugs supported by the carrier.

Description

[Technical Field]

[0001] [Cross-reference of related applications] This application claims priority based on Chinese application number 202310871629.2 (filing date: July 14, 2023). The disclosures of the aforementioned Chinese application are incorporated herein by reference in their entirety.

[0002] This invention relates to the field of pharmaceuticals, and more specifically to targeted chemical drugs, methods for producing the same, pharmaceutical compositions, and uses of targeted chemical drugs. [Background technology]

[0003] Conventional oncological chemotherapy is based on the principle that tumor cells are more susceptible to being killed by anticancer chemicals because they proliferate more rapidly. However, representative cytotoxic chemotherapeutic agents such as doxorubicin, paclitaxel, and cisplatin cannot distinguish between cancer cells and normal cells, and this lack of selectivity can lead to negative effects such as "killing a thousand enemies and destroying eight hundred of oneself," or even worse. In particular, high doses of cytotoxic drugs intended to eradicate tumors expose patients to the risk of systemic cytotoxicity. Therefore, a system that recognizes the inherent differences between tumor cells and normal cells and delivers them specifically is urgently needed for efficient cancer treatment. The development and accumulation of technology in nucleic acid nanocarriers through multi-targeting has provided the possibility of achieving specific delivery, but the reliability of delivery of small molecule chemicals by existing nucleic acid nanocarriers still needs further improvement. [Overview of the project]

[0004] The primary objective of the present invention is to provide targeted chemical drugs, methods for producing them, pharmaceutical compositions, and applications for targeted chemical drugs, thereby solving the following problems in the prior art: targeted delivery of small molecule chemical drugs, reduction of toxicity, enhancement of efficacy, and expansion of the therapeutic window.

[0005] To achieve this objective, according to one aspect of the present invention, a targeted chemical drug is provided. The drug comprises a targeted nucleic acid carrier and a small molecule chemical drug supported on the targeted nucleic acid carrier. The targeted nucleic acid carrier comprises a nucleic acid carrier and a target molecule linked to the nucleic acid carrier. Here, the nucleic acid carrier is a DNA carrier or an RNA carrier and includes sequences A, B, and C that form a self-assembling structure, where sequence A is sequence number 1 (SEQ ID NO. 1): 5'-ACGAGCGTTCCG-3'; sequence B is sequence number 2 (SEQ ID NO. 2): 5'-CGGTTCGCCG-3'; sequence C is sequence number 3 (SEQ ID NO. 3): 5'-CGGCCATAGCCGT-3'; or sequence A is sequence number 4 (SEQ ID NO. 4): 5'-ACGAGCGUUCCG-3'; sequence B is sequence number 5 (SEQ ID NO. 5): 5'-CGGUUCGCCG-3'; and sequence C is sequence number 6 (SEQ ID NO. 6): 5'-CGGGCCAUAGCCGU-3'. Optionally, at least one base of sequences A, B, and C may be substituted, inserted, or deleted.

[0006] According to a second aspect of the present invention, a pharmaceutical composition is provided comprising the target chemical drug and a pharmaceutically acceptable carrier and / or excipient.

[0007] According to a third aspect of the present invention, the above-mentioned target chemical drug is provided for use in the manufacture of a pharmaceutical product for the treatment of tumors.

[0008] Furthermore, the cancerous tumor may be gastric cancer and / or lung cancer. Preferably, the route of administration is intratumoral, intravenous, or intraperitoneal.

[0009] A fourth aspect of the present invention provides a method for preventing and / or treating cancer. The method includes the steps of providing the target chemical drug or pharmaceutical composition and administering an effective amount of the target chemical drug or pharmaceutical composition to a patient with a tumor.

[0010] Furthermore, tumors may be urogenital tumors, gynecological tumors, respiratory tumors, digestive tumors, hematological tumors, urinary tract tumors, bone marrow tumors, neurological tumors, dermatological tumors, general surgical tumors, or endoscopic tumors. More preferably, urogenital tumors are prostate cancer, penile cancer, testicular tumors, or male urethral cancer; gynecological tumors are ovarian cancer, cervical cancer, endometrial cancer, uterine fibroids, vulvar cancer, or malignant hydatidiform mole; respiratory tumors are lung cancer, non-small cell lung cancer, small cell lung cancer, nasopharyngeal cancer, tracheal tumors, lung metastases, inflammatory pseudotumor, or radiation-induced lung cancer; digestive tumors are liver cancer, gastric cancer, colorectal cancer, gallbladder cancer, esophageal cancer, rectal cancer, pancreatic cancer, or colon cancer; hematological tumors are leukemia, phosphorus Pathocyanitic cancer, lymphosarcoma, or multiple myeloma; urinary tract tumors include renal cancer, bladder cancer, or urinary tract cancer; bone marrow tumors include giant cell tumor, osteochondroma, or osteosarcoma; neurological tumors include brain tumors, meningiomas, cerebral tuberculomas, pituitary tumors, neuroblastomas, glioblastomas, or gliomas; dermatological tumors include skin cancer or malignant melanoma; general surgical tumors include breast cancer, lipoma, thyroid cancer, or thyroid tumors; and tumors of the five organs may include oral cancer, tongue cancer, laryngeal cancer, middle ear cancer, gingival cancer, or orbital tumors. Preferably, the route of administration is intratumorial, intravenous, or intraperitoneal, and the daily dose is preferably 0.1 μg / kg to 100 mg / kg.

[0011] A fifth aspect of the present invention provides a method for producing the targeted chemical drug, the method comprising the steps of: forming a targeted nucleic acid carrier having an arbitrary oligonucleotide effector molecule and an arbitrary immunostimulant by a sequence self-assembly mode; and ligating a small molecule chemical drug to the targeted nucleic acid carrier to obtain the targeted chemical drug.

[0012] The target chemical drug according to the present invention comprises a targeted nucleic acid carrier and a small molecule chemical drug carried by the targeted nucleic acid carrier. The targeted nucleic acid carrier comprises a nucleic acid carrier and a target molecule linked to the nucleic acid carrier. Here, the nucleic acid carrier is a DNA carrier or an RNA carrier, and comprises sequences A, B and C that form a self-assembled structure. On one hand, due to the coating effect of the above DNA carrier, the small molecule chemical drug can be embedded in the carrier to prevent its dropping during the delivery process. On the other hand, by modifying the carrier with a target aptamer, it has excellent targeting properties compared with other nucleic acid carriers. Furthermore, a combined therapeutic effect is exhibited between the small molecule chemical drug and other drugs carried by the carrier. Generally speaking, the present target chemical drug can deliver the small molecule chemical drug more stably and reliably, and exhibits a therapeutic effect by the combination of the small molecule chemical drug and other drugs carried by the carrier.

Brief Description of the Drawings

[0013] The drawings in the specification constituting a part of this application are for providing a further understanding of the present invention. The exemplary embodiments and descriptions of the present invention are for explaining the present invention and shall not unduly limit the present invention. In the drawings:

[0014] [Figure 1] Shows the measurement results by HPLC chromatogram (SEC) of the crude product of the targeted nucleic acid carrier after sequence assembly in Example 6 of the present invention; [Figure 2] Shows the measurement results by HPLC chromatogram (SEC) of the targeted nucleic acid carrier after HPLC purification in Example 6 of the present invention; [Figure 3] Shows the measurement results by HPLC chromatogram (reverse phase) of the crude product of the target chemical drug carrying epirubicin in Example 6 of the present invention; [Figure 4] Shows the measurement results by HPLC chromatogram (reverse phase) of the crude product of the target chemical drug after HPLC purification in Example 6 of the present invention; [Figure 5]The HPLC chromatogram (reverse phase) of the target chemical drug obtained by changing the molar ratio of epirubicin to carrier to 10:1 in Example 6 of the present invention is shown; [Figure 6] The HPLC chromatogram (reverse phase) of the target chemical drug obtained by changing the molar ratio of epirubicin to carrier to 20:1 in Example 6 of the present invention is shown; [Figure 7] The HPLC chromatogram (reverse phase) of the target chemical drug obtained by changing the molar ratio of epirubicin to carrier to 30:1 in Example 6 of the present invention is shown; [Figure 8] The relative growth curve of the body weight of each group of mice in the in vivo efficacy test of the N87 gastric cancer mouse model for the epirubicin target drug Apt AS1411 - Apt EGFR - Apt A15 - 2*Bio - DNA - epirubicin prepared in Example 5 of the present invention is shown; [Figure 9] The tumor growth curve of each group of mice in the in vivo efficacy test of the N87 gastric cancer mouse model for the epirubicin target drug Apt AS1411 - Apt EGFR - Apt A15 - 2*Bio - DNA - epirubicin prepared in Example 5 of the present invention is shown; [Figure 10] The change in tumor volume of each group of mice in the in vivo efficacy test of the N87 gastric cancer mouse model for the epirubicin target drug Apt AS1411 - Apt EGFR - Apt A15 - 2*Bio - DNA - epirubicin prepared in Example 5 of the present invention is shown; [Figure 11] The change curve of mouse body weight in the comparison of the in vivo efficacy test of the N87 gastric cancer mouse model for the epirubicin target drug Apt AS1411 - Apt EGFR - Apt A15 - 2*Bio - DNA - epirubicin prepared in Example 5 of the present invention and the epirubicin target drug Apt AS1411 - Apt EGFR - Apt A15 - 2*Bio - AmiR - 21 - DNA - epirubicin prepared in Example 7 is shown; [Figure 12]The curves showing the change in mouse tumor volume are shown in a comparison of in vivo efficacy studies of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin prepared in Example 5 of the present invention and the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin prepared in Example 7 in an N87 gastric cancer mouse model; [Figure 13] The relative tumor volume change curves in mice are shown in a comparison of in vivo efficacy studies of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin prepared in Example 5 of the present invention and the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin prepared in Example 7 in an N87 gastric cancer mouse model; [Figure 14] The curve showing the change in mouse tumor volume in an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-4*Bio-DNA-epirubicin, prepared in Example 3 of the present invention, is shown. [Figure 15] The curves showing the relative tumor volume change in mice in an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-4*Bio-DNA-epirubicin, prepared in Example 3 of the present invention, are shown below. [Figure 16] The curves showing the change in mouse tumor volume in an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin, prepared in Example 6 of the present invention, and the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin, prepared in Example 20, are shown below. [Figure 17]The curves showing the relative tumor volume changes in mice in an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin, prepared in Example 6 of the present invention, and the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin, prepared in Example 20, are shown below. [Figure 18] The curve showing the change in mouse tumor volume in an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin, prepared in Example 6 of the present invention, is shown below. [Figure 19] The relative tumor volume change curves in mice during an in vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin, prepared in Example 6 of the present invention, are shown below. [Figure 20] The tumor images of groups 1, 2, and 4 in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin, prepared in Example 9 of the present invention, are shown below; [Figure 21] The following shows the body weight change curves for each group of mice in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention; [Figure 22] The percentage change in body weight of each group of mice in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1), prepared in Example 8 of the present invention, is shown below; [Figure 23]The following are in vivo imaging images of the dorsal side of mice in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention; [Figure 24] The image shows the left-side in vivo imaging of a mouse in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention; [Figure 25] The image shows a right-sided in vivo imaging of a mouse in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention; [Figure 26] The image shows a ventral in vivo imaging of a mouse model of GL261-Luc brain glioma in an in vivo drug efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention; [Figure 27] The following shows the curves of change in bioluminescence imaging (BLI) values ​​in mice during an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 of the present invention, with (a) being the BLI value change curve on the dorsal side of the mouse, (b) being the BLI value change curve on the left side of the mouse, (c) being the BLI value change curve on the right side of the mouse, and (d) being the BLI value change curve on the ventral side of the mouse; [Figure 28] The survival curves for each group of mice in an in vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1), prepared in Example 8 of the present invention, are shown. [Modes for carrying out the invention]

[0015] The embodiments and features described herein can be combined with each other, provided they do not contradict each other. The present invention will be described in detail below with reference to the drawings, along with the embodiments.

[0016] Explanation of terms: Oligonucleotides refer to oligonucleotide molecules, including small interfering RNAs (siRNAs), antisense nucleic acids (ASOs), microRNAs (miRNAs), and nucleic acid aptamers (Aptamers). Oligonucleotide drugs are composed of nucleotides and represent a novel category of pharmaceuticals that is entirely different from small molecule drugs and antibody drugs. Compared to conventional chemical drug molecules, oligonucleotide drugs have advantages such as high target specificity, high efficiency, sustained efficacy, ease of drug design, and a wide range of candidate targets. The main oligonucleotide drugs are siRNA drugs and antisense nucleic acid drugs, both of which primarily target mRNA in the cytoplasm and achieve the objective of disease treatment by controlling protein expression through base complementary recognition and suppression of target mRNA. In this application, "nucleic acid" or "small nucleic acid" both refer to small nucleic acids.

[0017] Antisense nucleic acids (ASOs): ASOs, or antisense oligonucleotides, are typically short (16-53 nucleotides) synthetic oligonucleotides used to inhibit the function of RNA (including mRNA or miRNA). ASOs generally have high complementarity with target RNA and, structurally, contain chemically modified nucleotides or have specific sequences added to both ends to increase their affinity for the target sequence and resist nucleolysis within cells.

[0018] When targeting mRNA, ASOs are designed to form complementary pairs with specific regions of the target mRNA, thereby blocking translational lead generation, inducing mRNA degradation, or altering mRNA splicing.

[0019] When targeting miRNAs, ASOs (also called anti-miRNA molecules depending on their function) are designed to form complementary pairs with specific miRNAs, thereby inhibiting the binding of miRNAs to their target mRNAs and suppressing miRNA function. (miRNAs are a type of short non-coding RNA that form complementary pairs with the 3' untranslated region (3'UTR) of target mRNAs, influencing mRNA stability and translation, and further regulating protein expression.) In this application, ASOs that function as anti-miRNA molecules include, but are not limited to, A-miR-21, A-miR-10a, A-miR-30c, and AmiR1306.

[0020] MicroRNAs (miRNAs) are short, non-coding RNAs approximately 22 nucleotides long. Their mechanism of action involves miRNAs forming complementary pairs with the 3' untranslated region (3'UTR) of target mRNA, thereby regulating the stability or translation of the target mRNA and ultimately influencing protein expression. miRNAs have been shown to play crucial roles in many biological processes, including development, differentiation, proliferation, and apoptosis.

[0021] miRNAs have diverse forms, the earliest being pri-miRNA. Pri-miRNA undergoes primary processing to become pre-miRNA (microRNA precursor), which then undergoes further cleavage by the Dicer enzyme to become mature miRNA. In actual research, the use of pre-miRNA was the earliest and most widespread, and many commercially available microRNA libraries are in the pre-miRNA form. In recent years, it has been discovered that both arms of microRNA play a crucial role in the formation of mature miRNA, and the natural pri-miRNA form is increasingly being adopted by researchers.

[0022] Furthermore, as part of a therapeutic strategy, the concept of miRNA supplements has been proposed to increase the levels of specific miRNAs (e.g., miR-34a or miR-122) in the body. These supplements are typically analogs or mimics of miRNAs and may be in RNA or DNA form. If in DNA form, they can function similarly to normal miRNAs within the cell, as long as the DNA can be transcribed to form RNA with the same sequence as a mature miRNA.

[0023] There are several advantages to using DN sequence A-form miRNA supplements. Firstly, DNA is more stable and more resistant to degradation enzymes in the body. Secondly, because DNA is read by the cell's transcription mechanism and transcribed into the corresponding RNA, it can increase intracellular miRNA levels. Finally, the difficulty and cost of DNA synthesis are relatively low, and DNA is easy to design and manufacture, especially in large-scale production and application.

[0024] Taking miR-34a as an example, it has been proven to play an important role in antitumor activity in many types of cancer, and increasing the levels of miR-34a in the body can be an effective cancer treatment strategy. By administering miR-34a supplements in the DN sequence A form (i.e., miRNA analogs), the levels of miR-34a in the body can be increased, thereby suppressing the proliferation and survival of cancer cells.

[0025] Therefore, the meaning of miRNA in this application is broad and includes the above-mentioned pri-miRNA, pre-miRNA, mature miRNA, and miRNA analogs (which are in DNA form and can form oligonucleotides with the same sequence and function as mature miRNA through transcription). In this application, miRNA analogs include, but are not limited to, one or more of miR-34, miR-542, miR-126, and miR-122.

[0026] While various carriers, including nucleic acid carriers, exist in existing technology to enhance drug delivery efficiency, the problem of limitations on the clinical application of drugs remains difficult to solve. Furthermore, although drug combination therapy is a common method in clinical treatment, it usually refers to the combination of single agents, and the technical difficulty of incorporating multiple effector molecules into the drug itself is extremely high. To solve these problems, the present invention provides a small molecule chemical drug comprising a targeted nucleic acid carrier and a small molecule chemical drug mounted on the targeted nucleic acid carrier. The targeted nucleic acid carrier comprises a nucleic acid carrier and a target molecule bound to the nucleic acid carrier, and the nucleic acid carrier is a DNA carrier or an RNA carrier and includes sequences A (sequence a), B (sequence b), and C (sequence c) that form a self-assembly structure. Sequence A may be sequence number (SEQ ID NO. 1): 5'-ACGAGCGTTCCG-3'; sequence B may be sequence number 2: 5'-CGGTTCGCCG-3'; sequence C may be sequence number 3: 5'-CGGCCATAGCCGT-3'; or sequence A may be sequence number 7: 5'-ACGAGCGUUCCG-3'; sequence B may be sequence number 8: 5'-CGGUUCGCCG-3'; and sequence C may be sequence number 9: 5'-CGGGCCAUAGCCGU-3'. Optionally, at least one of sequences A, B, and C may have a base substitution, insertion, or deletion.

[0027] This invention employs a special nucleic acid carrier, which not only maintains resistance to degradation by nucleases inherent to DNA molecules but also overcomes the problem of difficulty in self-assembly due to the rigid structure of conventional DNA. The DNA carrier is formed by the self-assembly of three pure single strands of DNA. Similarly, the RNA carrier also exhibits superior self-assembly performance. The inventors have further discovered that a small molecule chemical drug can be loaded onto the nucleic acid carrier, and after the target molecule is bound, it can be inserted into the carrier's CG structure in the form of a covalent bond, or bound to the carrier sequence in the form of a base substitution or extension, or linked to the carrier by a linker via a covalent bond. As a result, the small molecule chemical drug temporarily loses its activity and, during the delivery process, mainly exists loaded onto the carrier, either not detaching or existing only in a small amount as a free form, thus significantly reducing toxicity. On the other hand, the carrier is mediated by the target molecule, particularly by a complex target aptamer, to more effectively target target tissues, the target tissue microenvironment, and target cells, entering the target cells through endocytosis or intracellular uptake (including pinocytosis). There, under the synergistic effects of a low intracellular pH environment, reduction by glutathione, and enzymatic cleavage, it releases the small molecule chemical drug, achieving highly efficient targeting intervention for the small molecule chemical drug and significantly improving its bioavailability. Simultaneously, when the carrier carries other drugs (e.g., gene modulators, immunomodulators) and the small molecule chemical drug together, the small molecule chemical drug effectively generates synergistic effects with other drug effector molecules, significantly increasing the efficiency of monotherapy and reducing toxicity.

[0028] In one preferred embodiment, the 5' and 3' ends of sequences A, B, and C are independently ligated with extended base portions, each containing 0 to 14 bases. Preferably, the nucleic acid carrier includes the following sequences that form a self-assembling structure: First set of sequences: Sequence A (a-sequence):SEQ ID NO. 7:5'-GCGGCGCCACGAGCGTTCCGGGAGC-3'; Sequence B (b-sequence):SEQ ID NO. 8:5'-GCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C (c-sequence):SEQ ID NO. 9:5'-GGCGGCAGGCGGCCATAGCCGTGGGCGCCGC-3'; or Second set of sequences: Sequence A:SEQ ID NO. 10:5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 11: 5'-GCTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C:SEQ ID NO. 12:5'-CGCGGCTCGCGGCCATAGCCGTGGGCGCCGC-3'; or Third set of sequences: Sequence A:SEQ ID NO. 10:5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B:SEQ ID NO. 13:5'-GCTCCTCTCCCGGTTCGCCGCCAGCCGCGG-3'; Sequence C:SEQ ID NO. 14:5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The fourth set of sequences: Sequence A:SEQ ID NO. 15:5'-GCGGCGCCACCGAGCGTTCCGGGAGAGGCC-3'; Sequence B:SEQ ID NO. 16:5'-GGCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C:SEQ ID NO. 14:5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or Fifth set of sequences: Sequence A:SEQ ID NO. 17:5'-GCGGCGCCCACGAGCGUUCCGGGAGAGGCC-3'; Sequence B:SEQ ID NO. 18:5'-GGCCUCUCCCGGUUCGCCGCCAGCCGCC-3'; Sequence C:SEQ ID NO. 19:5'-GGCGGCUGGCGGCCAUAGCCGUGGGCGCCGC-3'; someone The sixth sequence group: Sequence A:SEQ ID NO. 20:5'- GCGGCGCCCACGAGCGTTCCGGGAGAGGAGGCC-3'; Sequence B:SEQ ID NO. 21:5'- GGCCTCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C:SEQ ID NO. 14:5'- GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The seventh sequence group: Sequence A:SEQ ID NO. 10:5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 22: 5'-GCTCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C:SEQ ID NO. 14:5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The eighth sequence group: Sequence A:SEQ ID NO. 23:5'-GCGACGCCCACGAGCGTTCCGGGAGAGGAG-3'; Sequence B: SEQ ID NO. 24: 5'- CTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C:SEQ ID NO. 25:5'- CGCGGCACGCGGCCATAGCCGTGGGCGTCGC-3'; or The ninth sequence group: Sequence A:SEQ ID NO. 26:5'-GCGACGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 11: 5'-GCTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C:SEQ ID NO. 27:5'- CGCGGCTCGCGGCCATAGCCGTGGGCGTCGC-3'; or The 10th set of sequences: Sequence A: SEQ ID NO. 28: 5'- GACGCCCACGAGCGTTCCGGGAGAGG-3'; Sequence B:SEQ ID NO. 29:5'- CCTCTCCCGGTTCGCCGCGAGCCT-3'; Sequence C:SEQ ID NO. 30:5'-GGCTCGCGGCCATAGCCGTGGGCGTCTGCTGCTGCTGCTG -3'; or The 11th set of sequences: Sequence A: SEQ ID NO. 31: 5'- GCCCACGAGCGTTCCGGGAGA-3'; Sequence B: SEQ ID NO. 32: 5'- TCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C:SEQ ID NO. 33:5'- GGCGGCTGGCGGCCATAGCCGTGGGC-3'.

[0029] The nucleic acid carriers formed by self-assembly using the above-mentioned sequence groups can exert the aforementioned effects even more effectively.

[0030] As described above, when the nucleic acid carrier according to the present invention is used to simultaneously carry other pharmaceuticals (e.g., gene modifiers, immunomodulators) and small molecule chemicals, the small molecule chemicals produce extremely good synergistic effects of combined drug efficacy with the other pharmaceutical effector molecules, significantly improving the efficiency of monotherapy and reducing toxicity. Specifically, the targeted chemical may further include oligonucleotide effector molecules and / or immunostimulants carried on the targeted nucleic acid carrier. Preferably, the oligonucleotide effector molecule comprises one or more of ASO, siRNA, miRNA, and nucleic acid aptamers; more preferably, the siRNA comprises one or more of ASAP1, ATAD2, CD24, CD47, EGFR, HBV, HSP, HS70, PD-L1, PAPP-1, Survivin, TAP, TIM-3, TGF-β1 (TGF-beta1), and VEGF-C; even more preferably, the siRNA comprises one or more of ASAP1, CD47, PD-L1, and TGF-β1; more preferably, the miRNA comprises one or more of A-miR21, A-miR-10a, A-miR-30c, miR-34, miR-542, miR-126-3p, and miR-122.More preferably, the miRNA is A-miR21; more preferably, the nucleic acid aptamer comprises a nucleic acid aptamer in DNA form and / or a nucleic acid aptamer in RNA form, with preferred nucleic acid aptamers being A1, A15, AS1411, AFP, ATP, Act-12c, A18, BAF7-1, C-Met-SL1, CH6, CA2, C12, and CRAC. Orail, CEA, CEA-18, CEA-T84, CSC1, CSC13, CD40, CD16a, CD19, CD3-4, CD44, CD12(HDLBP), CD20, CD24, CD33, CD38, CD105, CD117, CD 63, CD123, EGFR, EpCAM, EcR, FAP, anti-FAP, GPC-1, GSK836, GPC3(APS63-1), Her2, Her3, HMGA2, H2, HbsAg, IFN-y(B4), IL-4Ra, IL-17, LZ H8, MUC1, M5, M7, M1, N5, NG-Dua, NKG2D(20-N-15), NSE, Np-A15, Np-A48, Np-A58, Np-A61, OX40, PSMA, PDGFRβ (PDGFRbeta), PDGF, PD -L1, PD-1, PTK-7, ProGRP-48, SF, TBA15, TBA29, TfRA4, TfRA3, TTA1, TLS9a, TGF-βII(S58)(TGF-betaII(S58)), TNF-a, TNF, T1, VEGF Examples include one or more of the following: VCAM-1, VCAM-12d, CH6, PL-45, EP66, AGC, Karpas299, SW620, MDA-MB-231, MCF-7, PC-3, BCMA, CTLA-4, CCL1, CD4-3, CD28, FGF2(F2), FGF2, FGF5, LAG-3, MRP1, TIM3, TIMC-11, VEGF165, and 4-1BB. More preferably, the nucleic acid aptamers include one or more of the following: A1, A15, AS1411, C12, CD40L, EGFR, GPC-1, IL-4Ra, MUC1, OX40, PD-L1, TTA1, TfRA3, and TfRA4. Preferably, the immunostimulant comprises one or more of the following: CPG2006, CPG1826, CPG2216, CPG2395, CPG-ODNT7, and CPG-ODN-PCIF1. More preferably, the immunostimulant is CPG2006.

[0031] Specifically, the sequence structure of the sense strand and antisense strand of each siRNA from the 5' end to the 3' end is as follows: ASAP1: The sense strand is SEQ ID NO. 34:5'-UGAUAUUAUGGAAGCAAAUUU-3', and the antisense strand is SEQ ID NO. 35:5'-AUUUGCUUCCAUAAUAUCAUU-3'; or, the sense strand is SEQ ID NO. 36:5'-UUAGGUUUGGGGUUGGAUCUU-3', and the antisense strand is SEQ ID NO. 37:5'-GAUCCAACCCCAAACCUAAUU-3'; ATAD2: The sense strand is SEQ ID NO. 38:5'-AAUCCUACAACUUCGACGCUU -3', and the antisense strand is SEQ ID NO. 39:5'-GCGUCGAAGUUGUAGGAUUUU-3'; CD24: The sense strand is SEQ ID NO. 40:5'-UGUUUACAUUGUUGAGGUAUU-3', and the antisense strand is SEQ ID NO. 41:5'-UACCUCAACAAUGUAAACUUU-3'; CD47: The sense strand is SEQ ID NO. 42:5'-GGUGAUUACCCAGAGAUAUTT-3', the antisense strand is SEQ ID NO. 43:5'-AUAUCUCUGGGUAAUCACCTT-3'; or the sense strand is SEQ ID NO. 44:5'-UGGUGAAAGAGGUCAUUCCUU-3', the antisense strand is SEQ ID NO. 45:5'-GGAAUGACCUCUUUCACCAUU-3'; or the sense strand is SEQ ID NO. 46:5'-GGAAUGACCUCUUUCACCATT-3', the antisense strand is SEQ ID NO. 47:5'-UGGUGAAAGAGGUCAUUCCTT-3'; or the sense strand is SEQ ID NO. 48:5'-GGUGAUUACCCAGAGAUAUUU-3', the antisense strand is SEQ ID NO. 49:5'-AUAUCUCUGGGUAAUCACCUU-3'; Preferably, the base modifications in CD47 are as follows: phosphorothioate (PS) modification between two adjacent bases from the 2nd to 4th bases from the 5' end of the sense strand, PS modification between two adjacent bases from the 1st to 3rd bases from the 3' end of the sense strand, PS modification between two adjacent bases from the 1st to 2nd bases from the 5' end of the antisense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 3' end of the antisense strand; the purpose of the above base modifications is to further improve the structural stability of the drug, and all base modifications that appear below are for this purpose, so no redundant explanation will be given.

[0032] EGFR: The sense strand is SEQ ID NO. 50:5'-GGCUGGUUAUGUCCUCAUUUU-3', and the antisense strand is SEQ ID NO. 51:5'-AAUGAGGACAUAACCAGCCUU-3'; or the sense strand is SEQ ID NO. 52:5'-UUAGAUAAGACUGCUAAGGUU-3', and the antisense strand is SEQ ID NO. 53:5'-UUAGAUAAGACUGCUAAGGUU-3'; or the sense strand is SEQ ID NO. 54:5'-UGCCUUAGCAGUCUUAUCUAAUU-3', and the antisense strand is SEQ ID NO. 55:5'-UUAGAUAAGACUGCUAAGGCAUU-3'; or the sense strand is SEQ ID NO. 78:5'-UGCCUUAGCAGUCUUAUCUAAUUUU-3' The antisense strand is SEQ ID NO. 79:5'- AAUUAGAUAAGACUGCUAAGGCAUU-3'; HBV: Its sense strand is SEQ ID NO. 56: 5'-GGACUUCUCUCAAUUUUCUUU-3', and its antisense strand is SEQ ID NO. 57: 5'-AGAAAAUUGAGAGAAGUCCUU-3'; preferably, the HBV has 2'-fluoro(2'-F) modifications on C and U; HSP: The sense strand has SEQ ID NO. 58: 5'-CGCAGAACACCGUGUUCGAUU-3', and the antisense strand has SEQ ID NO. 59: 5'-UCGAACACGGUGUUCUGCGUU-3'; preferably, in the HSP, PS modification is made between two adjacent bases from the 1st to 3rd bases from the 5' end of the sense strand, and PS modification is made between two adjacent bases from the 1st to 3rd bases from the 3' end of the antisense strand; HS70: The sense strand is SEQ ID NO. 60: 5'-GGCCAACAAGAUCACCAUC-3', and the antisense strand is SEQ ID NO. 61: 5'-GAUGGUGAUCUUGUUGGCCUU-3'; preferably, in the HS70, PS modification is made between two adjacent bases from the 1st to 3rd bases from the 5' end of the antisense strand, and PS modification is made between two adjacent bases from the 1st to 3rd bases from the 3' end; PD-L1: Its sense strand is SEQ ID NO. 62:5'-CCAGCACACUGAGAAUCAAUU-3', and its antisense strand is SEQ ID NO. 63:5'-UUGAUUCUCAGUGUGCUGGUU-3'; or its sense strand is SEQ ID NO. 64:5'-AGACGUAAGCAGUGUUGAATT-3', and its antisense strand is SEQ ID NO. 65:5'-UUCAACACUGCUUACGUCUTT-3'; PAPP-1: Its sense strand is SEQ ID NO. 66:5'-GAGGAAGGUAUCAACAAAUTT-3', and its antisense strand is SEQ ID NO. 67:5'-AUUUGUUGAUACCUUCCUCTT-3'; Survivin: The sense strand is SEQ ID NO. 70:5'-GCAGGUUCCUUAUCUGUCACAUU-3', and the antisense strand is SEQ ID NO. 71:5'-UGUGACAGAUAAGGAACCUGCAGUU-3'; or, The sense strand has SEQ ID NO. 72:5'-GGAAUUGGAAGGCUGGGAACCUU-3', and the antisense strand has SEQ ID NO. 73:5'-GGUUCCCAGCCUUCCAAUUCCUU-3'; preferably, PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the sense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the antisense strand, and PS modification between the 1st to 2nd bases from the 3' end; or, The sense strand is SEQ ID NO. 74:5'-UGCAGGUUCCUUAUCUGUCATT-3', and the antisense strand is SEQ ID NO. 75:5'-UGACAGAUAAGGAACCUGCTT-3'; or, The sense strand has SEQ ID NO. 76:5'-CUGCAGGUUCCUUAUCUGUCACAUU-3', and the antisense strand has SEQ ID NO. 77:5'-UGUGACAGAUAAGGAACCUGCAGUU-3'; preferably, the sense strand has PS modification between two adjacent bases from the 5' end to the 1st to 3rd bases, and PS modification between the 1st to 3rd bases from the 3' end, and the antisense strand has PS modification between two adjacent bases from the 5' end to the 1st to 3rd bases, and PS modification between the 1st to 3rd bases from the 3' end; TAP: The sense strand has SEQ ID NO. 80: 5'-GCUGCACACGGUUCAGAAUUU-3', and the antisense strand has SEQ ID NO. 81: 5'-AUUCUGAACCGUGUGCAGCUU-3'; preferably, PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the sense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the antisense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 3' end; or, The sense strand is SEQ ID NO. 82:5'-CAGGAUGAGUUACUUGAAAUU -3', and the antisense strand is SEQ ID NO. 83:5'- UUUCAAGUAACUCAUCCUGUU -3'; TIM-3: Its sense strand is SEQ ID NO. 86:5'-GUGCUCAGGACUGAUGAAATT-3', and its antisense strand is SEQ ID NO. 87:5'-UUUCAUCAGUCCUGAGCACTT-3'; TGF-β1: Its sense strand is SEQ ID NO. 88:5'-GUCAACUGUGGAGCAACACUU-3', and its antisense strand is SEQ ID NO. 89:5'-GUGUUGCUCCACAGUUGACUU-3'; or its sense strand is SEQ ID NO. 90:5'-GCAACAACGCCAUCUAUGATT-3', and its antisense strand is SEQ ID NO. 91:5'-UCAUAGAUGGCGUUGUUGCTT-3'; VEGF-C: The sense strand has SEQ ID NO. 92:5'-GCAAGACGUUGUUUGAAAUUAUU-3', and the antisense strand has SEQ ID NO. 93:5'-UAAUUUCAAACAACGUCUUGCUU-3'; preferably, PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the sense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 5' end of the antisense strand, and PS modification between two adjacent bases from the 1st to 3rd bases from the 3' end; or, The sense strand is SEQ ID NO. 84:5'-CAGCAAGACGUUGUUUGAAAUUAUU -3', and the antisense strand is SEQ ID NO. 85:5'- UAAUUUCAAACAACGUCUUGCUGUU -3'; or, The sense strand has SEQ ID NO. 94:5'-CAGGAUGGUAAAGACUACAUU-3', and the antisense strand has SEQ ID NO. 95:5'-UGUAGUCUUUACCAUCCUGUU-3'; preferably, PS modification is applied between two adjacent bases from the 1st to 3rd bases from the 5' end of the sense strand, and PS modification is applied between two adjacent bases from the 1st to 3rd bases from the 5' end of the antisense strand, and PS modification is applied between two adjacent bases from the 1st to 3rd bases from the 3' end; Preferred miRNAs include one or more of the following: miR-34, miR-542, miR-126-3p, and miR-122; A preferred ASO comprises one or more of A-miR21, A-miR-10a, A-miR-30c, and A-miR-1306; The sequence structure of each miRNA and ASO from the 5' end to the 3' end is as follows: A-miR21: 5'-GATAAGCT-3'; preferably, each base is modified with locked nucleic acid (LNA); or, 5'-GAUAAGCU-3'; preferably, each base is modified with LNA; or, SEQ ID NO. 96:5'-GTCAACATCAGTCTGATAAGCTA-3'; or, SEQ ID NO. 97:5'-GUCAACAUCAGUCUGAUAAGCUA-3'; or, SEQ ID NO. 98:5'-TCAACATCAGTCTGATAAGCTA; or, 5'-GATAAGCT-3'; preferably, PS modification between two adjacent bases; A-miR-10a:5'-ACAGGGTA-3'; preferably, each base is modified with LNA; A-miR-30c:SEQ ID NO. 99:5'-GCTGAGAGTGTAGGATGTTTACA-3'; miR-34:5'-TGTGACAG-3'; miR-542:5'-TGGCAGTGT-3'; miR-126-3p:5'-UCGUACC-3'; preferably, PS modification between adjacent bases; miR-122:5'-GGAAGTGT-3; AmiR-1306:SEQ ID NO. 100:5'-CATCACCACCAGAGCCAACGTC-3'; preferably, a PS modification between two adjacent bases from the 1st to 5th bases from its 5' end; The sequence structure of each nucleic acid aptamer from the 5' end to the 3' end is as follows: A1:SEQ ID NO. 101:5'-GGTTGCATGCCGTGGGGAGGGGGGTGGGTTTTATAGCGTACTCAG-3'; A15: SEQ ID NO. 102: 5'-CCCTCCTACATAGGG-3'; or SEQ ID NO. 227: 5'-CCCUCCUACAUAGGG-3'; AS1411:SEQ ID NO. 103:5'-GGTGGTGGTGGTTGTGGTGGTGGTGG-3'; or SEQ ID NO. 228:5'-GGUGGUGGUGGUUGUGUGGUGGUGGG-3'; or SEQ ID NO. 229:AUUCUGAACCGUGUGCAGCACCACGCUGCACACGGUUCAGAAUACACACGAGGCTATCTAGAATGTAC-3'; AFP:SEQ ID NO. 104:5'-GGCAGGAAGACAAACAAGCTTGGCGGCGGGAAGGTGTTTAAATTCCCGGGTCTGCGTGGTCTGTGGTGCTGT-3'; or SEQ ID NO. 105:5'-ACCTGGGGAGTATTGGGGAGGAAGG-3'; ATP:SEQ ID NO. 106:5'-ACCTGGGGAGTATTGGGGAGGAAGG-3'; or SEQ ID NO. 107:5'-GGGAGGACGATGCGGAGGAAGGGTAGG-3', preferably with PS modification between two adjacent bases from the 5' end to the 1st to 4th bases; Act-12c:SEQ ID NO. 108:5'-CGGGGAAAGTCACGGGGGGTTTCAGATGTTCTGATCGGTGTGGAG-3'; A18:SEQ ID NO. 109:5'-CCAGATAGTCCCTGG-3'; BAF7-1:SEQ ID NO. 110:5'-GATAACGGGCACGAATTCGGAGTG-3'; C-Met-SL1:SEQ ID NO. 111:5'-ATCAGGCTGGATGGTAGCTCGGTCGGGGTGGGTGGGTTGGCAAGTCTGAT-3'; CH6:SEQ ID NO. 112:5'-AGTCTGTTGGACCGAATCCCGTGGACGCACCCTTTGGACG-3'; CA2:SEQ ID NO. 113:5'-CCCACGTCTGCGCTTAGCTCCTGGGCCTGGATGGGC-3'; C12: SEQ ID NO. 114: 5'-GTGGATTGTTGTGTTCTGTTGGTTTTTGTGTTGTC-3'; preferably, PS modification between two adjacent bases from the 5' end to the 1st to 3rd bases; CRAC Orail:SEQ ID NO. 115:5'-CCAGTAGCCATACCGGTTTGTGGATGGGGTGTATGCGAGT-3'; CEA: SEQ ID NO. 116:5'-CTAGGATCCCCCACTCACCATCTCTCAGCTTGCTTCCTAGC-3'; or, SEQ ID NO. 234:5'-CUAGGAUCCCCACUCACCAUCUCUCAGCUUGCUUCCUAGC-3';CEA-18:SEQ ID NO. 117: 5'-TTAACTTATTCGACCATA-3'; CEA-T84: SEQ ID NO. 118: 5'-TCGCGCGAGTCGTCTGGGGAACCATCGAGTTACACCGACCTTCTATGTGCGGCCCCCCGCATCGTCCTCCC-3'; CSC1: SEQ ID NO. 119:5'-ACCTTGGCTGTCGTGTTGTAGGTGGTTTGCTGCGGTGGGCTCAAGAAGAAAGCGCAAAGAGGTCAGTGGTCAGAGCGT-3'; CSC13: SEQ ID NO. 120:5'-ACCTTGGCTGTCTGTTGTGGGGTGTCGTATCTTTCGTGTCTTATTATTTCTAGGTGGAGGTCAGTGGTCAGAGCGT-3'; CD40: SEQ ID NO. 121:5'-CCAACGAGTAGGCGATAGCGCGTGG-3'; or SEQ ID NO. 122: 5'-AGAGACGATGCGGCCAACGAGTAGGCGATAGCGCGTGGCAGAGCGTCGCT-3'; CD16a: SEQ ID NO. 123: 5'-CCACTGCGGGGGTCTATACGTGAGGAAGAAGTGG-3'; CD19:SEQ ID NO. 124:5'-TGCGTGTGTAGTGTGTGTCGTTTCTCCTTTTTTTGGTTGCTGCTCTGGGATTTGGGCGG-3'; or, SEQ ID NO. 125:5'-TGCGTGTGTAGTGTGTGTCTGTTCTCCTTTTTTTTGGTTGCTGCTCTTAGGGATTTGGGCGG-3'; CD3-4:SEQ ID NO. 126:5'-TCTCGGACGCGTGTGGTCGGCCGAGTGGCCCACGGTAGAAGGGTTAGAACTGCTGGTTGGTGAATCTCGCTGCCTGGCCCTAGAGTG-3'; CD44: SEQ ID NO. 127:5'-CCAAGGCCTGCAAGGGAACCAAGGACACAG-3'; or SEQ ID NO. 128: 5'-CCAAGGCCTGCAAGGGAACCAAGGACACAG-3'; preferably, PS modification between two adjacent bases from the 3rd to 5th bases from the 5' end, PS modification between two adjacent bases from the 12th to 14th bases from the 5' end, PS modification between the 1st to 2nd bases from the 3' end, PS modification between the 3rd to 4th bases from the 3' end, PS modification between the 5th to 6th bases from the 3' end, PS modification between two adjacent bases from the 12th to 14th bases from the 3' end; or, SEQ ID NO. 237:5'-GGGAUGGAUCCAAGCUUACUGGCAUCUGGAUUUGCGCGUGCCAGAAUAAAGAGUAUAAACGUGUGAAUGGGAAGCUUCGAUAGGAAUUCGG-3'; CD12(HDLBP):SEQ ID NO. 129:5'-GTGGATTGTTGTGTTCTGTTGGTTTTTGTGTTGTC-3'; CD20:SEQ ID NO. 130:5'-TGCGTGTGTAGTGTGTCTGTTTTTTATCTTCTTTTATCTACTCTTAGGGATTTGGGCGG-3'; CD24:SEQ ID NO. 131:5'-TATGTGGGTGGGTGGGCGGTTATGCTGAGTCAGCCTTGCT-3'; or SEQ ID NO. 132:5'-ATCCAGAGTGACGCAGCATATGTGGGTGGGTGGGCGGTTATGCTGAGTCAGCCTTGCTTGGACACGGTGGCTTAGT-3'; CD33: SEQ ID NO. 133: 5’-TACCAGTGCGATGCTCAGCACGCTTATAGGGGCTGGACAAAATTCTACCCAGCCTTT-3’; CD38: SEQ ID NO. 134: 5’-TACGTGAATCTCGTACGATACTCTGTAAGCGT-3’; CD105: SEQ ID NO. 135: 5’-GATCAGTTTTCCATGCCAGTTGGTATTCCGCGACAGTTTGATCTC-3’; CD117: SEQ ID NO. 136: 5’-GGGGCCGGGGCAAGGGGGGGGTACCGTGGTAGGAC-3’; CD63: SEQ ID NO. 137: 5’-CACCCCACCTCGCTCCCGTGACACTAATGCTA-3’; CD123: SEQ ID NO. 138: 5’-TGCGTGTGTAGTGTGTCTGGGCTACATCGATGAGCTGCCTAGGGTCCCTCTTAGGGATTTGGGCGG-3’; EGFR: SEQ ID NO. 139: 5’-GCCTTAGTAACGTGCTTTGATGTCGATTCGACAGGAGGC-3’; or, SEQ ID NO. 239: 5’-GCCUUAGUAACGUGCUUUGAUGUCGAUUCGACAGGAGGC-3’; or, SEQ ID NO. 240: 5’-UGCCGCUAUAAUGCACGGAUUUAAUCGCCGUAGAAAAGCAUGUCAAAGCCGUU-3’; EpCAM: SEQ ID NO. 140: 5’-CACTACAGAGGTTGCGTCTGTCCCACGTTGTCATGGGGGGTTGGCCTG-3’; or, SEQ ID NO. 141: 5’-GACAAACGGGGGAAGATTTGACGTCGACGAC-3’; or, SEQ ID NO. 241: 5’-GCGACUGGUUACCCGGUCG-3’; EcR: SEQ ID NO. 142: 5’-GCAGGTCCACTGCGGGGGTCTATACGTGAGGAAGAAGTGGGCAGGTC-3’; FAP:SEQ ID NO. 143:5'-TGGGGGTTGAGGCTAAGCCGA-3'; Anti-FAP:SEQ ID NO. 144:5'-CCGCTCGAGCTAGTCTGACAAAGAGAAACAC-3'; GPC-1:SEQ ID NO. 145:5'-AACGGAGTGTGGCTAACTCGA-3'; GSK836:SEQ ID NO. 147:5'-GCAGAGGTGAAGCGAAGTCG-3'; preferably, a PS modification between two adjacent bases from the 1st to 6th bases from the 5' end; GPC3(APS63-1):SEQ ID NO. 148:5'-TAACGCTGACCTTAGCTGCATGGCTTTACATGTTCCA-3'; preferably, PS modification between two adjacent bases from the 1st to 4th positions from the 5' and 3' ends; Her2:SEQ ID NO. 149:5'-AGCCGCGAGGGGAGGGATAGGGTAGGGCGCGGCT-3'; or SEQ ID NO. 150:5'-GGGAGCTCAGAATAAACGCTCAAAGGGTCAAGCTGATTACACTTTGTCCACTATTGGGTCCTTCGACATGAGGCCCGGATC-3'; or SEQ ID NO. 246:5'-AGCCGCGAGGGGAGGGAUAGGGUAGGGCGCGGCU-3'; Her3:SEQ ID NO. 151:5'-GGGAGCTCAGAATAAACGCTCAAGGCTAACAGCACGCAACGGGGGGAGTAATCGTGTCTGTTCGACATGAGGCCCGGATC-3'; or SEQ ID NO. 247:5'-CAGCGAAAGUUGCGUAUGGGUCACAUCGCAGGCACAUGUCAUCUGGGCG-3'; or SEQ ID NO. 248:5'-GAAUUCCGCGUGUGCCAGCGAAAGUUGCGUAUGGGCCACAUCGCAGGCACAUGUCAUCUGGGCGGUCCGUUCGGGAUCC-3'; HMGA2:SEQ ID NO. 152:5'-GGAAAAAATTTTTTAAAAAACCC-3'; preferably, phosphorothioate (PS) modification between adjacent two bases; H2:SEQ ID NO. 146:5'-GGGCCGTCGAACACGAGCATGGTGCGTGGACCTAGGATGACCTGAGTACTGTCC-3'; HBsAg:SEQ ID NO. 153:5'-CACAGCGAACAGCGGCGGACATAATAGTGCTTACTACGAC-3'; preferably, PS modification between two adjacent bases from the 1st to 4th bases from the 5' end; IFN-y(B4):SEQ ID NO. 154:5'-CCGCCCAAATCCCTAAGAGAAGACTGTAATGACATCAAACCAGACACACACACTACACACGCA-3'; IL-4Ra:SEQ ID NO. 155:5'-GGAGGACGAUGCGGAAAAAGCAACAGGGUGCUCCAUGCGCAUGGAACCUGCGGCGCAGACGACUCGCUGAGGAUCCGAGA-3'; or, SEQ ID NO. 156:5'-AAAAAGCAACAGGGUGCUCCAUGCGCAUGGAACCUGCGCG-3'; IL-17:SEQ ID NO. 157:5'-CTTGGATCACCATAGTCGCTAGTCGAGGCT-3'; or, SEQ ID NO. 158:5'-GCGGCATCCTATCACGCATTGACC-3'; LZH8:SEQ ID NO. 159:5'-ATCCAGAGTGACGCAGCATATTAGTACGGCTTAACCCCATGGTGGACACGGTGGCTTAGT-3'; MUC1: SEQ ID NO. 160: 5’-GCAGTTGATCCTTTGGATACCCTGG-3’; or, SEQ ID NO. 161: 5’-GAAGTGAAAATGACAGAACACAACA-3’; or, SEQ ID NO. 162: 5’-AACCGCCCAAATCTCTAAGAGTCGGACTGCAACCTATGCTATCGTTGATGTCTGTCCAAGCAACACAGACACACTACACACACGCACA-3’; or, SEQ ID NO. 163: 5’-AATGACAGAACACAACATT-3’; or, SEQ ID NO. 250: 5’-GCAGUUGAUCCUUUGGAUACCCUGG-3’; M5: SEQ ID NO. 164: 5’-AGCAGCACAGAGGTCAGATGCTTGGTTCCACCGTACTGACTGTAGTAAAATCTGATCACTCCTATGCGTGCTACCGTGAA-3’; M7: SEQ ID NO. 165: 5’-AGCAGCACAGAGGTCAGATGTAGTCGGTCTTCTTGTTTGAAACTGCTAATTTTGAAAAAACCTATGCGTGCTACCGTGAA-3’; M1: SEQ ID NO. 166: 5’-AGCAGCACAGAGGTCAGATGATATAACCTTAATAAATAAAATATAAATTATTTAATCTTACCTATGCGTGCTACCGTGAA-3’; N5: SEQ ID NO. 167: 5’-GATTGAGTAGATAGTGGTTCTGTACGTAGTGAAAGAGTGG-3’; N-G-Dua: SEQ ID NO. 168: 5’-CAAGTTGCTCGTCGCGATACGTTTGGTTGGTGTGGTTGGCAGTATCGCAGGTCCAAGTTGCTCGTCGCGATACAACGGAGTGTGGCTAACTCGA-3’; NKG2D(20-N-15): SEQ ID NO. 169: 5’-CAAGTTGCTCGTCGCGATACGTTTGGTTGGTGTGGTTGGCAGTATC-3’; NSE:SEQ ID NO. 170:5’-TCACACGGACCTCTCTCTACATTAATTGCGCATTTCGTT-3’; Np-A15:SEQ ID NO. 171:5’-GCTGGATGTTCATGCTGGCAAAATTCCTTAGGGGCACCGTTACTTTGACACATCCAGC-3’; Np-A48:SEQ ID NO. 172:5’-GCTGGATGTCGCTTACGACAATATTCCTTAGGGGCACCGCTACATTGACACATCCAGC-3’; Np-A58:SEQ ID NO. 173:5’-GCTGGATGTCACCGGATTGTCGGACATCGGATTGTCTGAGTCATATGACACATCCAGC-3’; Np-A61:SEQ ID NO. 174:5’-GCTGGATGTTGACCTTTACAGATCGGATTCTGTGGGGCGTTAAACTGACACATCCAGC-3’; OX40: SEQ ID NO. 175: 5’-GGGAGGACGATGCGGCAGTCTGCATCGTAGGAATCGCCACCGTATACTTTCCCACCAGACGACTCGCTGAGGATCCGAGA-3’; or, SEQ ID NO. 176: 5’-CAGTCTGCATCGTAGGATTAGCCACCGUATCTTTCCCAC-3’; or, SEQ ID NO. 177: 5’-CCAACGAGTAGGCGATAGCGCGTGG-3’; or, SEQ ID NO. 252: 5’-GGGAGGACGAUGCGGCAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCACCAGACGACUCGCUGAGGAUCCGAGA-3’; or, SEQ ID NO. 253: 5’-GGGAGGACGAUGCGGCAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCACCAGACGACUCGCUG-3’; or, SEQ ID NO. 254: 5’-CAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCAC-3’; or, SEQ ID NO. 255: 5’-GGGAUGCGGAAAAAAGAACACUUCCGAUUAGGGCCCACCCUAACGGCCGCAGAC-3’; PSMA: SEQ ID NO. 179: 5’-GGGAGGACGATGCGGATCAGCCATGTTTACGTCACTCCT-3’; or, SEQ ID NO. 180: 5’-GCGTTTTCGCTTTTGCGTTTTGGGTCATCTGCTTACGATAGCAATGCT-3’; or, SEQ ID NO. 256: 5’-GGGACCGAAAAAGACCUGACUUCUAUACUAAGUCUACGUUCCC-3’; or, SEQ ID NO. 257: 5’-GGGAGGACGAUGCGGAUCAGCCAUGUUUACGUCACUCCU-3’; PDGFRβ: SEQ ID NO. 181: 5’-TGTCGTGGGGCATCGAGTAAATGCAATTCGACA-3’; or, SEQ ID NO. 258: 5’-UGUCGUGGGGCAUCGAGUAAAUGCAAUUCGACA-3’; PDGF: SEQ ID NO. 182: 5’-CAGGCTACGGCACGTAGAGCATCACCATGATCCTG-3’; PD-L1: SEQ ID NO. 183: 5’-AACAACGTATAACAATGCCCACGTCACCAGAGTACTATGG-3’; or, SEQ ID NO. 184: 5’-GCCCCAGTTATGCTTTCCCCCTCTGTCTCTTTG-3’; or, SEQ ID NO. 185: 5’-ATCGCCCGCAGCACCCATTTGTTTTTTTTTG-3’; or, SEQ ID NO. 69: ACGGGCCACATCAACTCATTGATAGACAATGCGTCCACTGCCCGT; or, SEQ ID NO. 186: 5’-TGCCCGCACATCAACTCATTGATAGACAATGCGTCCACTCGGGCA-3’; or, SEQ ID NO. 187: 5’-TGCCCGCACATCAACTCATTGATAGACAATGCGTCCACTACGGGC-3’; or, SEQ ID NO. 188: 5’-CGGGCACACATCAACTCATTGATAGACAATGCGTCCACTGCCCGT-3’; or, SEQ ID NO. 189: 5’-GTTGGTCACATCAACTCATTGATAGACAATGCGTCCACTACCAAC-3’; or, SEQ ID NO. 190: 5’-GGGCCACATCAACTCATTGATAGACAATGCGTCCACTGCCC -3’; or, SEQ ID NO. 191: 5’-TGGTTGCACATCAACTCATTGATAGACAATGCGTCCACTCAACCA-3’; or, SEQ ID NO. 200: 5’-TACAGGTTCTGGGGGGTGGGTGGGGAACCTGTT-3’; or, SEQ ID NO. 238: 5’-CTACGAGACGAACTTATGCGTAATAATGACTGTCGTAG-3’; PD-1: SEQ ID NO. 192: 5’-GACGATAGCGGTGACGGCACAGACGGTACAGTTCCCGTCCCTGCACTACACGTATGCCGCTTCCGTCCGTCGCTC-3’; or, SEQ ID NO. 193: 5’-GAGCGACGGACGGAAGCGGCATACGTGTAGTGCAGGGACGGGAACTGTACCGTCTGTGCCGTCACCGCTATCGTC-3’; or, SEQ ID NO. 194: 5’-GGATCCTAGACGCATTGACCCGCTGCCTCTACTGAGGCTGTGTCAGTGTGCGGCTCGGACTGTTGAATTC-3’; or, SEQ ID NO. 195: 5’-AGCGGTGACGGCACAGACGGTACAGTTCCCGTCCCTGCACTACACGTATGCCGCTGAGAGAGAGGGAGGC-3’; or, SEQ ID NO. 196: 5’-ACCGACAGTGAAGGACTCAGCGAACTCTCAGACTCGGTTC-3’; PTK-7: SEQ ID NO. 197: 5’-ATCTAACTGCTGCGCCGCCGGGAAAATACTGTACGGTTAGA-3’; ProGRP-48: SEQ ID NO. 198: 5’-CATGCGGAGTAGAGCGAGCCCAGATAGTCCCTGGTTATTTCCTTAGG-3’; SF: SEQ ID NO. 199: 5’-GATCTCTCTCTGCCCTAAGTCCGCACCCGTGCTTCCCTGT-3’; TBA15: SEQ ID NO. 201: 5’-GGTTGGTGTGGTTGG-3’; TBA29: SEQ ID NO. 202: 5’-AGTCCGTGGTAGGGCAGGTTGGGGTGACT-3’; TfRA4: SEQ ID NO. 203: 5’-GCGTGGTACCACGC-3’; or, SEQ ID NO. 259: 5’-GCGUGGUACCACGC-3’; TfRA3:SEQ ID NO. 204:5′-GCGTGGTCACACGC-3′; 205:5'-GCGGCGCCCACGAGCGTTCGCGTGGTCACACGCGTTCCGCCCTCCTACATAGGGCGCATAGCCGTGGGCGCCGC-3'; 260:5′-GCGUGGUCACACGC-3′; TTA1:SEQ ID NO. 206:5′-CCTGCACTTGGCTTGGATTTCAGAAGGGAGACCC-3′; 286:5'-CTGCACTTGGCTTGGATTTCAGAAGGGAGACCC-3'; 261:5′-CCUGCACUUGGCTTGGAUUUCAGAAGGGAGACCC-3′; TLS9a:SEQ ID NO. 207:5′-AGTCCATTTTATTCCTGAATATTTGTTAACCTCATGGAC-3′; TGF-βII(S58):SEQ ID NO. 208:5'-ACATTGCTGCGTGATCGCCTCACATGGGTTTGTCTGGTCGATTTGGAGGTGGTGGGTGGC-3'; TNF-a:SEQ ID NO. 209:5'-GCGGCCGATAAGGTCTTTCCAAGCGAACGAAAA-3'; TNF:SEQ ID NO. 210:5′-GCGCCACTACAGGGGAGCTGCCATTCGAATAGGTGGGCGC-3′; T1:SEQ ID NO. 211:5'-CGCTCGATAGATCGAGCTTCGCTCGATGTGGTGTTGTGGGGGCTTGTATTGGTCGATCACGCTCTAGAGCACTG-3'; VEGF: SEQ ID NO. 212: 5’-TGTGGGGGTGGACTGGGTGGGTACC-3’; or, SEQ ID NO. 213: 5’-TGTGGGGGTGGACGGGCCGGGTAGA-3’; or, SEQ ID NO. 214: 5’-GGTGGGGGTGGACGGGCCGGGTAGA-3’; or, SEQ ID NO. 266: 5’-AUGCAGUUUGAGAAGUCGCGCAU-3’; preferably, with PS modification between the 6th and 7th bases from the 5’ end; VCAM-1: SEQ ID NO. 215: 5’-ATACCAGCTTATTCAATTGGACACGGCAAAGGGGTATAGCCTACCGGACCGTGAACATGGAATGGTGTGCTGCGTGGAGATAGTAAGTGCAATCT-3’; or, SEQ ID NO. 216: 5’-GGACACGGCAAAGGGGTATAGCCTACCGGACCGTGAACATGGAATGGTGTGCTGCGTGG-3’; VCAM-12d: SEQ ID NO. 217: 5’-AGGGAATCTTGCCTAGGGAGGGAGTAGCGAAAGGGCTCA-3’; CH6: SEQ ID NO. 218: 5’-AGTCTGTTGGACCGAATCCCGTGGACGCACCCTTTGGACG-3’; PL-45: SEQ ID NO. 219: 5’-ACTCATAGGGTTAGGGGCTGCTGGCCAGATACTCAGATGGTAGGGTTACTATGAGC-3’; EP66: SEQ ID NO. 220: 5’-AACAGAGGGACAAACGGGGGAAGATTTGACGTCGACGACA-3’; AGC: SEQ ID NO. 221: 5’-CGACCCGGCACAAACCCAGAACCATATACACGATCATTCGTCTCCTGGGCCG-3’; Karpas299: SEQ ID NO. 222: 5’-ATCCAGATGACGCAGCACCACCACCGTACAATTTTTTCATTACCTACTCGGC-3’; SW620: SEQ ID NO. 223: 5’-CCCATCAATGTTACGACCCGCTAGGGCTGCTGTGCCATCGGGTAA-3’; MDA-MB-231: SEQ ID NO. 224: 5’-AGAATTCAGTCGGACAGCGAAGTAGTTTTCCTTCTAACCTAAGAACCCGCGGCAGTTTAATGTAGA-3’; MCF-7: SEQ ID NO. 225: 5’-GCATGGGGTTTCGGCGTTTCGTCTATCTTGTTTCTGTTAGCGTCT-3’; PC-3: SEQ ID NO. 226: 5’-TGCCACTACAGCTGGTTCGGTTTGGTGACTTCGTTCTTCGTTGTGGTGCTTAGTGGC-3’; BCMA: SEQ ID NO. 230: 5’-AGUGCAAGACGUUCGCAGAUUAGCGAAAAGAGGGUCUCAUUGACUAGUAC-3’; CTLA-4: SEQ ID NO. 231: 5’-GGUGGAAGAGGGAUGGGCCGACGUGCCGCAU-3’; or, SEQ ID NO. 232: 5’-TCCCTACGGCGCTAACGATGGTGAAAATGGGCCTAGGGTGGACGGTGCCACCGTGCTACAAC-3’; CCL1: SEQ ID NO. 233: 5’-UGACUCCUCUGACAGCCUAAUUUCUCCCGAUUACCCUG-3’; CD4-3: SEQ ID NO. 235: 5’-GGGAGGACGAUGCGGUUUGGGGUUUUCCCGUGCCCCAGACGACUCGCCCGA-3’; CD28: SEQ ID NO. 236: 5’-GGGAGAGAGGAAGAGGGAUGGGGAUUAGACCAUAGGCUCCCAACCCCCCCCGGGAGAGAGGAAGAGGGAUGGGGAUUAGACCAUAGGCUCCCAACCCCCGGG-3’; FGF2(F2):SEQ ID NO. 242:5'-GGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCC-3'; FGF2: SEQ ID NO. 243:5'-GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCUCGA-3'; or, SEQ ID NO. 244:5'-GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCCC-3'; FGF5:SEQ ID NO. 245:5'-GGGCGACCUCCUCCGUACUGACCUACAGAGCGACAUACUACUAGUGUAUCCAGAUCGCCC-3'; LAG-3: SEQ ID NO. 249:5'-GGGAGAGAGAUAUAAGGGCCUCCUGUAUACCCGCUGCUAUCUGGACCGAUCCCCAUUACCAAAUUCUCUCCCC-3'; MRP1: SEQ ID NO. 251:5'-GGGAGAAUAGUCAACAAAUCGUUUGGGGCGACUUCUCCUUCCUUUCUCCCC-3'; TIM3: SEQ ID NO. 262:5'-GGGAGAGGACCAGUA-GCCACUAUGGGUGUUGGAGCUAGCGG-CAGAGCGUCGCGGUCCCUCCC-3'; 263:5'-GGGAGAGGACCAGUA -CUGGUAGUUCUCUCUGUGCGACUCCUA-CAGAGCGUCGCGGUCCCUCCC-3'; TIMC-11: SEQ ID NO. 264:5'-AAGCAACACUUAGUCGCGAUUGAUACGUGCGCAGUCAU-3'; or, SEQ ID NO. 265:5'-GGAGGACGAUGCGGGGGAAGCAACACUUAGUCGCGAUUGAUACGUGCGCAGUCAUCAGACGACUCCGCUGAGGAUCCGAGA-3'; VEGF165: SEQ ID NO. 267:5'-CGGAAUCAGUGAAUGCUUA UACAUCCG-3'; 4-1BB:SEQ ID NO. 268:5'-GGGAGAGAGGAAGAGGGAUGGGCGACCGAACGUGCCCUUCAAAGCCGUUCACUAACCAGUGGCAUAACCCAGAGGUCGAUAGUACUGGAUCCCCC-3'; The sequence structure of each immunostimulant described above, from the 5' end to the 3' end, is as follows: CPG2006: SEQ ID NO. 269: 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3'; preferably, PS modification between adjacent two bases; or, SEQ ID NO. 270:5'-UCGUCGUUUUGUCGUUUUGUCGUU-3'; CPG1826:SEQ ID NO. 271:TCCATGACGTTCCTGACG-3'; preferably, PS modification between adjacent two bases; CPG2216:SEQ ID NO. 272:5'-GGGGGACGATCGTCGGGGGG-3'; CPG2395:SEQ ID NO. 273:TCGTCGTTTTCGGCGCGCGCCG-3'; CPG-ODNT7:SEQ ID NO. 274:5'-TCGTCGTCGTCGTCGTCGTCG-3'; or, SEQ ID NO. 275:5'-TCGTCGTCGTCGTCGTCGTCGTCG-3'; CPG-ODN-PCIF1:5'-AGCGAA-3'.

[0033] Preferably, if the target chemotherapy drug contains a nucleic acid aptamer, the target molecule contains a nucleic acid aptamer and / or a low-molecular-weight target ligand; if the target chemotherapy drug does not contain a nucleic acid aptamer, the target molecule is a low-molecular-weight target ligand, which is one or more selected from the group consisting of folic acid, biotin, vitamin B12, and mannose.

[0034] By loading the aforementioned types of oligonucleotide-effect molecules, immunostimulants, and small-molecule targeted ligands onto the specific nucleic acid carrier along with small-molecule chemical drugs, further synergistic effects can be achieved, leading to more beneficial results. For example, the nucleoline nucleic acid aptamer AS1411 binds specifically and with high affinity to nucleolins on the surface of cancer cells, while showing no affinity to normal cells. When AS1411 binds to nucleolins on the tumor cell membrane, it induces macropinocytosis and is rapidly taken into the cell, further promoting macropinocytosis by inducing the activity of EGFR, Akt, p38, and Rac1. Furthermore, nucleolins, as multifunctional shuttle proteins, shuttle between the cell membrane, cytoplasm, and cell nucleus during this process, functioning as "escort" molecular chaperones. EGFR-targeted aptamers are highly expressed in tumor cells as auxiliary target molecules for combined targeting. By using EGFR aptamers, they bind specifically and with high affinity to EGFR receptor proteins on the surface of cancer cells, while being unaffected or having low affinity to normal cells. CD133-positive (CD133+) tumor cells have stronger autoregeneration, differentiation, and tumorigenetic abilities. CD133 is a common marker for tumor stem cells and is highly expressed in many solid tumors (e.g., glioma, breast cancer, gastric cancer, liver cancer, lung cancer, pancreatic cancer, prostate cancer, colorectal cancer, etc.). As a CD133 aptamer, A15 has a targeting effect that tracks and specifically binds to CD133+ tumor cells in the body. In this invention, an improved chemical drug is constructed by modifying one end of the trifacial structure of the DNA carrier with an A15 aptamer by a sequence extension method, and more preferably by combining it with modified AS1411 and EGFR aptamers on the other two ends of the nanonucleic acid carrier to form a composite target, and loading epirubicin. This targeted improved epirubicin-targeted drug, when administered intravenously, anchors in multiple ways to tumor cells having nucleolin, EGFR, and CD133 that are overexpressed in tumor tissue, synergistically increasing the opportunity and efficiency of target binding between the target drug and tumor cells. After internalization and uptake into cells, the nanonucleic acid aggregate undergoes reduction and enzymatic cleavage reactions under low pH, high glutathione concentration, and nuclease action, releasing the epirubicin active ingredient.Epirubicin enters the cell nucleus, and its condensed tetracyclic structure (anthracycline nucleus) specifically intercalates at the GC / CG site of target gene DNA, inhibiting DNA transcription and thereby suppressing tumor growth and progression. Here, the A15 aptamer mediates further tracking of CD133+ tumor stem cell-like cells by nanonucleic acid aggregates, suppressing the growth of key cells that form the "seeds" of tumors, thereby controlling or reversing tumor progression. miR-21 is a gene that induces drug resistance, and by using its antisense sequence, the expression of resistance can be suppressed, increasing the cytotoxic effect of the drug and ultimately more effectively killing tumor cells. The TfRA4 targeted ligand mediates the passage of nanonucleic acid particles across the blood-brain barrier (BBB), thereby allowing them to carry epirubicin and more effectively kill brain tumor cells. In addition to AS1411, A15, EGFR, and TfRA4 aptamers, other target aptamers provided by the present invention can be linked to the DNA carrier to exert corresponding effects, thereby achieving a better delivery effect of epirubicin.

[0035] The low molecular weight chemical agents used in the present invention are of a type commonly used in the field, but are not limited to, anthracycline chemical agents, pyrimidine chemical agents, platinum chemical agents, glutamate derivative chemical agents, flavonoid chemical agents, plant-derived drugs and their derivatives, folic acid chemical agents, salicylic acid chemical agents, or acridine chemical agents. From the viewpoint of improving binding stability with nucleic acid carriers, in one preferred embodiment, the anthracycline chemical agent is selected from doxorubicin, epirubicin, pirarubicin, daunorubicin, idarubicin, mitoxantrone, barurubicin, or their free bases or hydrochlorides; preferably, the pyrimidine chemical agent is selected from gemcitabine, 5-fluorouracil, cytarabine, or capecitabine; preferably, the platinum chemical agent is selected from cisplatin, oxaliplatin, carboplatin, nedaplatin, or lovaplatin; preferably, the glutamate derivative chemical agent is selected from lenalidomide, thalidomide, or pomalidomide; preferably, the flavonoid chemical agent is flavone, flavo The chemical is selected from ol, dihydroflavone, isoflavone, or chalcone; preferably, the plant-derived chemical and its derivatives are selected from vincristine, dihydroartemisinin, paclitaxel, meitansine, docetaxel, or 10-hydroxycamptothecin; preferably, the folic acid chemical is selected from methotrexate; preferably, the salicylic acid chemical is selected from aspirin or sodium salicylate; preferably, the acridine chemical is selected from tacrine or aminacrine (aminoacridin); more preferably, the low molecular weight chemical is selected from one or more of epirubicin or its free base or hydrochloride, gemcitabine, 5-fluorouracil, paclitaxel, or meitansine.

[0036] Preferably, when a small molecular weight target ligand, oligonucleotide effector, and immunostimulant are linked to a nucleic acid carrier, they are linked directly to the terminal bases of each sequence on the nucleic acid carrier or via a base linker containing 1 to 10 bases; preferably, when the small molecular weight chemical is an anthracycline or acridine, the small molecular weight chemical is inserted between adjacent GC base pairs of the targeted nucleic acid carrier in the form of specific intercalation; when the small molecular weight chemical is a pyrimidine, the small molecular weight target ligand, oligonucleotide effector, and immunostimulant are linked to the terminal bases of each sequence on the nucleic acid carrier via a base linker containing 1 to 10 bases Linked to a base, the low molecular weight chemical is linked to the nucleic acid carrier by at least a portion of it substituting a base in the base linker and / or acting as an extended base and / or substituting a base in the carrier backbone; when the low molecular weight chemical is a platinum-based chemical, a glutamate derivative chemical, a flavonoid chemical, a plant-derived chemical and its derivatives, a folic acid-based chemical, or a salicylic acid-based chemical, the low molecular weight chemical is linked to the nucleic acid carrier by covalent bonding via the linker; the preferred linker is selected from long-chain primary amine compounds, more preferably a long-chain primary amine compound containing C2-C14, and even more preferably 5-bromomethyl-6-bromo-1-hexylamine (C7). After condensation with two molecules of phosphate ester, a base phosphoramidite monomer containing a linker is further prepared (for example, N4-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine-3'-(6-aminohexyl-2-hydroxymethyl)-N,N'-diisopropylphosphoramidite, or N2-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyguanosine-3'-(6-aminohexyl-2-hydroxymethyl)-N,N'-diisopropylphosphoramidite).

[0037] The loading amount of each of the above-mentioned small molecule chemicals can vary from 1 to the theoretical loading amount. Here, the theoretical loading amount refers to the maximum amount that can be theoretically loaded based on the sequence structure of the nucleic acid carrier. For example, when a small molecule chemical is inserted between adjacent GC base pairs of a targeted nucleic acid carrier in the form of specific intercalation, it will be understood by those skilled in the art that the number of adjacent GC base pairs in the sequence becomes the theoretical loading amount. In the actual loading process, the molar ratio of epirubicin to nucleic acid carrier is 2 to 300:1, preferably 2 to 290:1, more preferably 2 to 29:1, even more preferably 10 to 50:1, and most preferably 15 to 25:1.

[0038] For the purpose of further improving the size and / or stability of drugs during the delivery process, and for the purpose of optimizing the efficacy of drug combinations and reducing toxicity to enhance efficacy, in one preferred embodiment, the present invention provides the following more reliable targeted chemical drugs: (1) The target chemotherapy comprises a DNA carrier, a small molecule chemotherapy drug, and an EGFR nucleic acid aptamer linked to the DNA carrier, and optionally biotin, the preferred small molecule chemotherapy drug being epirubicin or its free base or hydrochloride; preferably, the DNA carrier comprises a first set of sequences, the EGFR nucleic acid aptamer is linked to the 5' end of sequence B, and the molar ratio of the EGFR nucleic acid aptamer to the DNA carrier is 1:1; preferably, the EGFR nucleic acid aptamer is linked to the 5' end of sequence B via a base linker TTTTT; when the target chemotherapy comprises a DNA carrier, a small molecule chemotherapy drug, and an EGFR nucleic acid aptamer linked to the DNA carrier and biotin, 2 to 4 biotin molecules are linked to each DNA carrier, and each biotin molecule is linked to the 5' or 3' end of sequence A or sequence C, respectively. Specifically, the target chemotherapy comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin mounted on the targeted nucleic acid carrier:

[0039] <Drug 1> TIFF2026522667000002.tif82164 Note: The bolded portion indicates sequence A, sequence B, or sequence C in the nucleic acid carrier. The same applies below.

[0040] In drug 1 described above, the nanonucleic acid carrier modifies the EGFR aptamer by a sequence extension method and specifically binds to the EGFR protein highly expressed in tumor tissue cells through active targeting. Approximately 30 epirubicin mounting sites for GC / CG are designed on the three double-stranded DNA arms formed by the self-assembly of three core skeleton sequences. In the manufacturing process, an excess ratio of 40:1 (epirubicin:nanonucleic acid carrier) is preferably used, and 4% paraformaldehyde is used in combination as a coupling agent. The condensed tetracyclic structure (anthracycline nucleus) of the epirubicin molecule specifically intercalates at the GC / CG sites of the nanonucleic acid carrier, and the amino group of its daunosamine structure forms a covalent bond reaction of Schiff bases with an amino group on one of the guanosine outer rings in GC / CG, which is the binding site of the carrier, under the catalyst of paraformaldehyde, thereby establishing a stable mounting mode through physical intercalation + covalent bonding. This drug not only possesses a natural passive targeting "funnel" accumulation effect on tumor tissue, but also has target affinity for tumor tissue and cells that highly express EGFR protein. Through the mediation of these two targeting effects, it can accumulate at high concentrations in tumor sites. After the EGFR aptamer binds to the EGFR protein highly expressed on the surface of tumor cells, it is taken into the cell via mechanisms such as endocytosis and reaches the cytoplasm. Under the influence of low pH and high concentrations of glutathione reducing agents within the tumor cells, epirubicin is dissociated and released from the nanocarrier. The released free epirubicin translocates to the nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA, blocking DNA transcription. Therefore, it inhibits the differentiation and proliferation of tumor cells, producing an antitumor effect that suppresses tumor development and progression. Specifically, the mechanisms of delivery, transport, release, sustained release, and inhibitory action of this drug are as follows:

[0041] 1. Delivery: EGFR-DNA + epirubicin nanoparticles are introduced into the body via a targeted delivery method, exhibiting a natural passive targeting "funnel" accumulation effect. 2. Transport: After the EGFR aptamer binds to the EGFR protein highly expressed on the surface of tumor cells, it is taken into the cell by mechanisms such as endocytosis. This binding causes the nanoparticles to accumulate at high concentrations in the tumor site, enhancing the drug's efficacy. 3. Release: Once the drug enters tumor cells, epirubicin is dissociated and released from the nano-nucleic acid carrier under the action of low pH and high concentration glutathione reducing agents. 4. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier via a stable loading mode consisting of physical interposition and covalent bonding, allowing for slow release and reducing drug side effects in the body. 5. Mechanism of inhibitory action: Released epirubicin enters the cell nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding blocks DNA transcription, suppressing the differentiation and proliferation of tumor cells and exerting an antitumor effect.

[0042] Based on the above, the delivery, transport, release, sustained release, and inhibitory mechanisms of drug 1 make it a promising targeted antitumor agent. By specifically targeting tumor tissues and cells with high EGFR protein expression, utilizing the passive targeting and accumulation effect of nanoparticles to effectively release the drug slowly, and specifically blocking the transcription of tumor cell DNA, this drug offers a new direction for research in tumor treatment.

[0043] <Drug 2> TIFF2026522667000003.tif80164

[0044] In drug 2 described above, the nano-nucleic acid carrier modifies the EGFR aptamer via a sequence extension method, and further links four biotin molecules to the ends of the other two arms via PEG modification to mediate active targeting, specifically binding to EGFR protein and biotin receptor protein highly expressed in tumor tissue cells. Approximately 30 epirubicin loading sites for GC / CG are designed on three double-stranded DNA arms formed by the self-assembly of three core skeletal sequences, thus establishing a stable loading mode via physical intercalation + covalent bonding. The mechanisms of delivery, transport, release, sustained release, and inhibitory action of this drug are as follows:

[0045] 1. Delivery: The nano-nucleic acid carrier achieves active targeting through modification of the EGFR aptamer and linkage to biotin. This structure allows the drug to specifically bind to EGFR protein and biotin receptor protein, which are highly expressed in tumor tissue cells. 2. Transport: After EGFR aptamers and biotin bind to target proteins on the surface of tumor cells, they are taken into the cells by mechanisms such as endocytosis. This binding causes the nanoparticles to accumulate at high concentrations in the tumor site, enhancing the drug's efficacy. 3. Release: Once the drug enters tumor cells, epirubicin is dissociated and released from the nano-nucleic acid carrier under the action of low pH and high concentration glutathione reducing agents. 4. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier via a stable loading mode consisting of physical interposition and covalent bonding, allowing for slow release and reducing drug side effects in the body. 5. Mechanism of inhibitory action: Released epirubicin enters the cell nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding blocks DNA transcription, suppressing the differentiation and proliferation of tumor cells and producing an antitumor effect.

[0046] Overall, the delivery, transport, release, sustained release, and inhibitory mechanisms of drug 2 make it a promising targeted antitumor agent. By specifically targeting tumor tissues and cells with high expression of EGFR protein and biotin receptor protein, utilizing the passive targeting and accumulation effect of nanoparticles to effectively release the drug slowly, and specifically blocking the transcription of tumor cell DNA, this drug offers new research directions in tumor treatment.

[0047] (2) The target chemical agent comprises a DNA carrier, a small molecular weight chemical agent, and a DNA-type nucleic acid aptamer AS1411, a nucleic acid aptamer EGFR, a DNA-type nucleic acid aptamer A15, and an optional biotin, wherein the preferred small molecular weight chemical agent is epirubicin or its free base or hydrochloride; preferably, the DNA carrier comprises a first set of sequences, with a molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier being 1:1, and linked to the 5' end of sequence A; preferably, the nucleic acid aptamer AS1411 is linked to the 5' end of sequence A via a base linker TTTTT; the molar ratio of nucleic acid aptamer EGFR to the DNA carrier is 1:1, and linked to the 5' end of sequence B ' Linked to the end; preferably, the nucleic acid aptamer EGFR is linked to the 5' end of sequence B via the base linker TTTTT; the molar ratio of the DNA-form nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is linked to the 5' end of sequence C; preferably, the nucleic acid aptamer A15 is linked to the 5' end of sequence C via the base linker TTTTT; if the target chemologic drug comprises a DNA carrier, a small molecule chemologic drug, and the DNA-form nucleic acid aptamers AS1411, EGFR, A15, and biotin linked to the DNA carrier, then two biotins are linked to each DNA carrier, and each biotin is linked to the 3' ends of sequences A and C, respectively. Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin mounted on the targeted nucleic acid carrier:

[0048] <Drug 3> TIFF2026522667000004.tif64163

[0049] <Drug 4> TIFF2026522667000005.tif63163

[0050] In drugs 3 and 4 described above, the nanonucleic acid carrier is modified by a sequence extension method, with AS1411 aptamer, EGFR aptamer, and A15 aptamer attached to the 5' ends of the three oligonucleotide sequences constituting the nucleic acid carrier backbone, respectively, and optionally with biotin Bio attached to the 3' ends of the a and c chains via PEG modification. A complex targeted nanonucleic acid carrier is formed that specifically binds to nucleolin, EGFR protein, stem cell-like CD133 receptor, and biotin receptor protein, and the epirubicin drug is covalently mounted on it. The drug is a nanonucleic acid carrier, and complex targeting is achieved by modification with AS1411 aptamer, EGFR aptamer, A15 aptamer, and biotin Bio. The mechanisms of drug delivery, transport, release, sustained release, and inhibitory action are as follows:

[0051] 1. Delivery: The nano-nucleic acid carrier achieves complex targeting through modification with AS1411 aptamer, EGFR aptamer, A15 aptamer, and any biotin Bio. This allows the drug to specifically bind to different types of tumor cells, achieving specific binding by utilizing the affinity between the aptamer and the corresponding tumor cell surface receptor. 2. Transport: After the aptamer binds to target receptors on the surface of tumor cells, it enters the cell via mechanisms such as the endoplasmic reticulum pathway. This binding causes the nanoparticles to accumulate at high concentrations in the tumor site, enhancing the drug's efficacy. 3. Release: Once the drug enters tumor cells, epirubicin is dissociated and released from the nano-nucleic acid carrier under the catalytic action of a low pH and high concentration of glutathione reducing agent. 4. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier via a stable loading mode consisting of physical interposition and covalent bonding, and its slow release can reduce drug side effects in the body. 5. Mechanism of inhibitory action: Released epirubicin enters the cell nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding blocks DNA transcription, suppressing the differentiation and proliferation of tumor cells and producing an antitumor effect.

[0052] Overall, the delivery, transport, release, sustained release, and inhibitory mechanisms of drug 3 make it a potentially powerful complex targeted antitumor agent. By specifically targeting different receptors in tumor tissue and cells, utilizing the passive targeting and accumulation effect of nanoparticles, effectively releasing the drug slowly, and specifically blocking the transcription of tumor cell DNA, this drug offers new research directions in tumor treatment.

[0053] (3) The targeted chemologic drug comprises a DNA carrier, a low molecular weight chemologic drug, and a DNA-type nucleic acid aptamer AS1411 and biotin linked to the DNA carrier, wherein the preferred low molecular weight chemologic drug is epirubicin or its free base or hydrochloride; preferably, the DNA carrier comprises a first set of sequences, the DNA-type nucleic acid aptamer AS1411 is linked to the 5' end of sequence B, and the molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier is 1:1; preferably, the nucleic acid aptamer AS1411 is linked to the 5' end of sequence B via a base linker TTTTT; each DNA carrier has 2 to 4 biotin molecules linked to it, each biotin molecule linked to the 5' or 3' end of sequence A or sequence C, respectively. Specifically, the targeted chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin mounted on the targeted nucleic acid carrier:

[0054] <Drug 5> TIFF2026522667000006.tif76164

[0055] In drug 5 described above, the nanonucleic acid carrier achieves active targeted specific binding through modification of the AS1411 aptamer and optional biotin linkage. The mechanisms of drug delivery, transport, release, sustained release, and inhibitory action are as follows:

[0056] 1. Delivery: The nanonucleic acid carrier achieves active targeting through modification of the AS1411 aptamer and optional biotin linkage. This structure allows the drug to specifically bind to ribonucleic acid (NCL) and biotin receptor proteins highly expressed in tumor tissue cells. 2. Transport: After the AS1411 aptamer and biotin bind to target proteins on the surface of tumor cells, they are taken into the cells by mechanisms such as endocytosis. This binding causes the nanoparticles to accumulate at high concentrations in the tumor site, enhancing the drug's efficacy. 3. Release: Once the drug enters tumor cells, epirubicin is dissociated and released from the nano-nucleic acid carrier under the catalytic action of a low pH and high concentration of glutathione reducing agent. 4. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier via a stable loading mode consisting of physical intercalation and covalent bonding, and its slow release can reduce drug side effects in the body. 5. Mechanism of inhibitory action: Released epirubicin enters the cell nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding blocks DNA transcription, suppressing the differentiation and proliferation of tumor cells and producing an antitumor effect.

[0057] Overall, the delivery, transport, release, sustained release, and inhibitory mechanisms of the above-mentioned drug 5 make it a promising targeted antitumor agent. By specifically targeting tumor tissues and cells with high expression of ribonucleic acid (NCL) and biotin receptor proteins, utilizing the passive targeting and accumulation effect of nanoparticles to effectively release the drug slowly, and specifically blocking the transcription of tumor cell DNA, the drug offers a new direction for research in tumor treatment.

[0058] (4) The targeted chemical drug comprises a DNA carrier, a small molecule compound drug, and nucleic acid aptamer A1 and nucleic acid aptamer A15 linked to the DNA carrier, preferably epirubicin, its free base or hydrochloride, as the small molecule compound drug. Preferably, the DNA carrier comprises a first sequence group, the molar ratio of nucleic acid aptamer A1 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, nucleic acid aptamer A1 is linked to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of nucleic acid aptamer A15 to the DNA carrier is 1:1 and it is linked to the 5′ end of sequence C. Preferably, nucleic acid aptamer A15 is linked to the 5′ end of sequence B by a base linker "TTTTT". Specifically, the targeted chemical drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0059] <Drug 6> TIFF2026522667000007.tif63164

[0060] In drug 6 described above, the nano-nucleic acid carrier modifies the A1 aptamer and A15 aptamer to achieve specific binding through active targeting. The mechanisms of drug delivery, transport, release, sustained release, and inhibitory action are as follows:

[0061] 1. Delivery: The nano-nucleic acid carrier achieves active targeting through modifications of the A1 and A15 aptamers. This structure allows the drug to specifically bind to tumor tissue cells, achieving specific binding by utilizing the affinity that the A1 and A15 aptamers have for the CD133 receptor on the tumor cell surface. 2. Transport: After the A1 and A15 aptamers bind to target receptors on the surface of tumor cells, they are taken into the cells by mechanisms such as endocytosis. This binding causes the nanoparticles to accumulate at high concentrations in the tumor site, enhancing the drug's efficacy. 3. Release: When the drug enters tumor cells, epirubicin dissociates from the nano-nucleic acid carrier and is released under a reducing environment such as low pH and high concentration of glutathione. 4. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier in a stable manner through physical intercalation and covalence bonding, enabling slow release and thereby reducing side effects in the body. 5. Mechanism of inhibitory action: Released epirubicin enters the nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding inhibits DNA transcription, suppressing the differentiation and proliferation of tumor cells and exhibiting an antitumor effect.

[0062] In short, the mechanisms of delivery, transport, release, sustained release, and inhibitory action of the above-mentioned drug 6 make this pharmaceutical a potentially targeted antitumor agent. Through specific targeting of tumor tissue and cells, utilization of the passive targeting and accumulation effect of nanoparticles, effective drug release and sustained release, and specific inhibition of tumor cell DNA transcription, this pharmaceutical offers new research directions in tumor treatment.

[0063] (5) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and a DNA-type nucleic acid aptamer AS1411, aptamer EGFR, DNA-type nucleic acid aptamer A15, optionally biotin, and oligonucleotide A-miR-21 linked to the DNA carrier, preferably epirubicin, its free base, or its hydrochloride as the small molecule compound drug. Preferably, the DNA carrier comprises a first sequence group, the molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably, nucleic acid aptamer AS1411 is linked to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of aptamer EGFR to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, nucleic acid aptamer EGFR is linked to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of the DNA-type nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is ligated to the 5′ end of sequence C. Preferably, the nucleic acid aptamer A15 is ligated to the 5′ end of sequence C by a base linker "TTTTT". The molar ratio of oligonucleotide A-miR-21 to the DNA carrier is 1:1, and it is ligated to the 3′ end of sequence C. Optionally, two biotin molecules are ligated to each DNA carrier, with each biotin molecule ligated to the 3′ end of sequence A and to the end of oligonucleotide A-miR-21 furthest from sequence C, respectively. Specifically, the targeted chemologic drug comprises a targeted nucleic acid carrier formed by self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0064] <Drug 7> TIFF2026522667000008.tif87164

[0065] <Drug 8> TIFF2026522667000009.tif62164

[0066] In drugs 7 and 8 described above, the nanonucleic acid carrier achieves a three-step targeting mediation effect through AS1411, EGFR, A15 aptamers, and optional biotin. Details of the function of each molecule and the three-step targeting mediation effect are as follows:

[0067] 1. AS1411 Aptamer: AS1411 is an oligonucleotide that specifically binds to nucleolin receptors on the surface of tumor cells. This binding allows the drug to be taken up by tumor cells, achieving targeted antitumor activity.

[0068] 2. EGFR Aptamers: EGFR (epidermal growth factor receptor) is overexpressed in many malignant tumor cells. Through EGFR aptamers, drugs specifically bind to EGFR-overexpressing tumor cells, increasing the concentration of the drug within the tumor tissue.

[0069] 3. A15 Aptamer: A15 is an aptamer that specifically binds to the CD133 receptor on tumor stem-like cells. Tumor stem-like cells are key to tumor recurrence and drug resistance, and the A15 aptamer allows drugs to target tumor stem-like cells.

[0070] 4. A-miR-21: A-miR-21 is an antitumor ASO (antisense oligonucleotide) against microRNA-21. MicroRNA-21 is overexpressed in many tumors and promotes tumor development and progression. A-miR-21 suppresses the function of microRNA-21 and inhibits tumor development and progression.

[0071] 5. Epirubicin: Suppresses the differentiation and proliferation of tumor cells by inhibiting DNA transcription.

[0072] Through a three-stage targeting-mediated effect, the above-mentioned drug achieves effective delivery, transport, release, sustained release, and inhibitory action. Particular emphasis is placed on suppressing tumor development and progression by targeting tumor stem-like cells. This complex targeting strategy is expected to improve drug efficacy and reduce side effects, offering a new direction for the treatment of various malignant tumors.

[0073] (6) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and DNA-type nucleic acid aptamers TfRA4, AS1411, and A15 linked to the DNA carrier, preferably epirubicin, its free base, or its hydrochloride as the small molecule compound drug. Preferably, the DNA carrier comprises a first sequence group, the molar ratio of DNA-type nucleic acid aptamer TfRA4 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably, the nucleic acid aptamer TfRA4 is linked to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, the nucleic acid aptamer AS1411 is linked to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of the DNA-type nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is ligated to the 5′ end of sequence C. Preferably, the nucleic acid aptamer A15 is ligated to the 5′ end of sequence C by a base linker "TTTTT". Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequence, and epirubicin supported on the targeted nucleic acid carrier:

[0074] <Drug 9> TIFF2026522667000010.tif62164

[0075] In drug 9 described above, the AS1411 aptamer, TfRA4 aptamer, and A15 aptamer are modified at the 5′ ends of the three oligonucleotide sequences constituting the nucleic acid carrier backbone, forming a complex targeted nano-nucleic acid carrier that specifically binds to nucleolin, transferrin [receptor] on the BBB, and CD133 receptor and biotin receptor proteins on tumor stem-like cells. Furthermore, epirubicin is covalently supported to form a targeted epirubicin drug. This nano-nucleic acid targeted drug specifically binds to transferrin [receptor (TfR)] on the BBB, crosses the BBB with the association of AS1411 to reach the brain parenchyma, binds to nucleolin receptors highly expressed on the surface of tumors such as glioblastoma, and specifically binds to tumor stem-like cells that highly express CD133 receptor protein, is taken up into the cell, and suppresses tumor development and progression. Specifically, this drug achieves a three-stage targeting-mediated mechanism of delivery, transport, release, sustained release, and inhibitory action. The functions of each molecule and the details of the three-step targeting mediation effect are as follows:

[0076] 1. Delivery: The nano-nucleic acid carrier, combined with the TfRA4 aptamer, specifically binds to transferrin receptors on the blood-brain barrier (BBB), helping the drug cross the BBB and enter the brain. 2. Transport: After the drug passes through the BBB, the AS1411 aptamer helps to specifically bind to nucleolin receptors highly expressed on the surface of colloidal tumors in the brain, thereby achieving targeting of colloidal tumor cells. 3. Release and Sustained Release: Epirubicin, covalently supported on a nanonucleic acid carrier, is gradually released under reducing conditions such as low pH and high concentration of glutathione after the drug enters tumor cells. This release mechanism enables sustained release, which can enhance drug efficacy while reducing side effects. 4. Mechanism of Inhibitory Action: The A15 aptamer allows the drug to specifically bind to tumor stem-like cells that highly express the CD133 receptor protein, enter the cells, and exert an inhibitory effect. This is extremely important for controlling cerebral colloid tumors, and tumor stem-like cells are key factors in tumor recurrence and drug resistance.

[0077] Based on the above, drug 9 specifically targets transferrin receptors, nucleolin receptors, and CD133 receptors on tumor stem-like cells in the blood-brain barrier (BBB) ​​through a three-stage targeting-mediated effect, achieving effective drug delivery, transport, release, sustained release, and inhibitory effects. Particular emphasis is placed on its targeting of tumor stem-like cells to suppress the development and progression of cerebral glioma. This complex targeting strategy is expected to improve drug efficacy and reduce side effects, providing a new direction for the treatment of cerebral glioma and other malignant tumors.

[0078] (7) The targeted chemical drug comprises a DNA carrier, a small molecule compound drug, and DNA-type nucleic acid aptamers TfRA4, AS1411, and A15 linked to the DNA carrier, the small molecule compound drug being epirubicin, its free base, or its hydrochloride. The DNA carrier comprises an eleventh sequence group, the molar ratio of DNA-type nucleic acid aptamer TfRA4 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably, the nucleic acid aptamer TfRA4 is linked to the 5′ end of sequence A by the transition base sequence SEQ ID NO. 276:5′-TTGCGGCGAGCGGCGA-3′, and further, the target chemotherapy contains the complementary sequence SEQ ID NO. 277:5′-TCGCCGCTCGCCGCTT-3′, which is complementaryly linked to the transition base sequence SEQ ID NO. 276:5′-TTGCGGCGAGCGGCGA-3′, with its 3′ end linked to the 5′ end of the nucleic acid aptamer TfRA4. The molar ratio of the DNA-form nucleic acid aptamer AS1411 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, the nucleic acid aptamer AS1411 is linked to the 5′ end of sequence B by the base linker "TTTTT". The molar ratio of the DNA-type nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is ligated to the 5′ end of sequence C. Preferably, the nucleic acid aptamer A15 is ligated to the 5′ end of sequence C by a base linker "TTTTT". Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequence, and epirubicin supported on the targeted nucleic acid carrier:

[0079] <Drug 10> TIFF2026522667000011.tif62164

[0080] Drug 10 is a variation of drug 9, and the difference from drug 8 is that the 14 base sequences before and after the TfRA4 aptamer (the transition base sequence TTGCGGCGAGCGGCGA and the complementary sequence TCGCCGCTCGCCGCTT) are completely complementary. Through a self-assembly mechanism, it forms a configuration that further extends the neck structure by one step via a pair of TT / TT intervals, thereby further improving the stability of the pharmaceutical structure and solving the problem of polymerization caused by the ease with which TfRA4 molecules self-assemble with each other.

[0081] (8) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and DNA-type nucleic acid aptamers AS1411, TfRA3, and A15 linked to the DNA carrier, preferably the small molecule compound drug being epirubicin, its free base, or its hydrochloride. Preferably the DNA carrier comprises a first sequence group, the molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably the nucleic acid aptamer AS1411 is linked to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of DNA-type nucleic acid aptamer TfRA3 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably the nucleic acid aptamer TfRA3 is linked to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of the DNA-type nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is ligated to the 5′ end of sequence C. Preferably, the nucleic acid aptamer A15 is ligated to the 5′ end of sequence B by a base linker "TTTTT". Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0082] <Drug 11> TIFF2026522667000012.tif62165

[0083] In the above drug 11, the nano-nucleic acid carrier is modified by sequence extension to add AS1411 aptamer, TfRA3 aptamer, and A15 aptamer to the 5′ ends of the three oligonucleotide sequences constituting the nucleic acid carrier backbone, forming a complex targeted nano-nucleic acid carrier that specifically binds to nucleolin, BBB transferrin, tumor stem-like cell CD133 receptor, and biotin receptor protein. This carrier covalently supports epirubicin to form a targeted epirubicin drug. This nano-nucleic acid targeted drug specifically binds to transferrin on the BBB, passes through the BBB with the conjugation of AS1411 to reach the brain parenchyma, binds to nucleolin receptors highly expressed on the surface of colloidal tumors, and specifically binds to tumor stem-like cells that highly express CD133 receptor protein, entering the cell and suppressing tumor development and progression. Specifically, drug 11 is a nano-nucleic acid carrier, and complex targeting is achieved by modification with AS1411 aptamer, TfRA3 aptamer, and A15 aptamer. The mechanisms of drug delivery, transfer, release, sustained release, and inhibitory action are as follows:

[0084] 1. Delivery: The nano-nucleic acid carrier, modified with AS1411 aptamer, TfRA3 aptamer, and A15 aptamer, specifically binds to nucleolin, BBB transferrin, and tumor stem-like cell CD133 receptors. This allows the drug to specifically bind to transferrin on the BBB, cross the BBB, and reach the brain parenchyma. 2. Three-stage targeting-mediated effect: a. First-stage targeting: The drug binds to transferrin on the blood-brain barrier (BBB) ​​via the TfRA3 aptamer and crosses the BBB with the conjugation of AS1411. b. Second-stage targeting: After reaching the brain parenchyma, the drug binds to nucleolin receptors highly expressed on the surface of colloidal tumors via the AS1411 aptamer. c. Third-stage targeting: The drug specifically binds to tumor stem-like cells that highly express the CD133 receptor protein via the A15 aptamer. 3. Transfer: After the aptamer binds to the target receptor on the surface of tumor cells, it enters the cell via mechanisms such as the endoplasmic reticulum pathway. 4. Release: Once the drug enters tumor cells, epirubicin is released from the nano-nucleic acid carrier under the catalytic action of a low pH and high concentration of glutathione reducing agent. 5. Sustained release: The epirubicin molecule is bound to the nano-nucleic acid carrier through a stable support mechanism involving physical intercalation and covalence linkage, enabling slow release and thereby reducing toxic side effects in the body. 6. Mechanism of Inhibitory Action: Released epirubicin enters the nucleus and specifically binds to and intercalates at the GC / CG site of nuclear double-stranded DNA. This binding blocks DNA transcription, suppressing the differentiation and proliferation of tumor cells and exhibiting an antitumor effect. In particular, by binding to tumor stem-like cells that highly express the CD133 receptor protein via the A15 aptamer, the drug enters and suppresses tumor stem cells, exerting a crucial mechanism of action for controlling cerebral colloid tumors.

[0085] Tumor stem cells are self-renewing and multipotent cells that play a crucial role in tumor growth, recurrence, and drug resistance. By targeting the CD133 receptor, which is highly expressed on tumor stem-like cells, drug 11 can more effectively target these cells, achieving the goal of eliminating the tumor's origin and suppressing recurrence.

[0086] Overall, the delivery, transfer, release, sustained release, and inhibitory action mechanisms of drug 11 are achieved through a three-stage targeted mediation effect, the key being the suppression of the development and progression of cerebral glioma by targeting tumor stem cells. This complex targeted strategy is expected to improve drug efficacy and reduce toxic side effects, offering a new direction for the treatment of other malignant tumors such as cerebral glioma.

[0087] (9) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and an immunostimulant CPG2006 linked to the DNA carrier, a nucleic acid aptamer CD40, and a nucleic acid aptamer PD-L1 in DNA form, preferably the small molecule compound drug being epirubicin, its free base, or its hydrochloride. Preferably the DNA carrier comprises the ninth sequence group, the molar ratio of the immunostimulant CPG2006 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably the immunostimulant CPG2006 is linked to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of the nucleic acid aptamer CD40 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably the nucleic acid aptamer CD40 is linked to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of the nucleic acid aptamer PD-L1 in DNA form to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence C. Preferably, the nucleic acid aptamer PD-L1 is linked to the 5′ end of sequence C by a base linker "TTTTTT". Specifically, the target chemotherapy comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequence, and epirubicin supported on the targeted nucleic acid carrier:

[0088] <Drug 12> TIFF2026522667000013.tif67165

[0089] Note: In sequence A, all phosphate bonds between adjacent bases at positions 1-24 of the 5′ end are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 1-3 of the 3′ end are also phosphorothioate modified. In sequences B and C, the phosphate bonds between adjacent bases at positions 1-3 of the 5′ and 3′ ends are all phosphorothioate modified.

[0090] In drug 12 described above, the nano-nucleic acid carrier integrates three aptamers (CPG2006, CD40, and PD-L1), enabling active combination immunotherapy and targeting of chemotherapy drugs to the tumor microenvironment. Details of the function of each effector molecule and the synergistic tumor suppressive effect resulting from the integrated targeting of the tumor microenvironment are as follows:

[0091] 1. CPG2006 Aptamer: CPG2006 is a CpG oligonucleotide that functions as an immunostimulant and can activate the immune system via the TLR9 receptor. In the tumor microenvironment, CPG2006 activates dendritic cells (DCs) and B cells, thereby promoting T cell activation and proliferation, and enhancing the anti-tumor immune response. 2. CD40 aptamers: CD40 is a membrane-bound protein that can bind to the CD40 receptor and activate dendritic cells and B cells. CD40L enhances the tumor microenvironment and strengthens the anti-tumor immune response by activating immune cells. 3. PD-L1 Aptamers: PD-L1 is an immunosuppressive molecule. In the tumor microenvironment, tumor cells express PD-L1 to suppress T cell activity and evade immune surveillance. By designing PD-L1 aptamers, pharmaceuticals can block the interaction between PD-L1 and the PD-1 receptor, restore T cell activity, and enhance the anti-tumor immune response. 4. Epirubicin: Suppresses the differentiation and proliferation of tumor cells by blocking DNA transcription.

[0092] By loading epirubicin onto an active combination immunotherapy agent consisting of three aptamers, this drug enables the combined targeting of the tumor microenvironment with both immunotherapy and classical chemotherapy. In the tumor microenvironment, the CPG2006 and CD40L aptamers activate dendritic cells and B cells, respectively, and promote T cell activation and proliferation; the PD-L1 aptamer blocks the interaction between PD-L1 and the PD-1 receptor, restoring T cell activity. These three aptamers act synergistically to enhance the anti-tumor immune response. Simultaneously, epirubicin, as a classical chemotherapy agent, blocks DNA transcription and suppresses the differentiation and proliferation of tumor cells. The combination of chemotherapy and immunotherapy makes tumor cells more easily recognized and eliminated by immune cells under the action of chemotherapy agents.

[0093] The tumor microenvironment is a complex cellular and molecular network comprising tumor cells, immune cells, supporting cells, and extracellular matrix. This drug achieves the synergistic tumor-suppressive effects of immunotherapy and chemotherapy by integrally targeting the tumor microenvironment. Firstly, the drug activates immune cells (e.g., dendritic cells, B cells, and T cells) to modulate the tumor microenvironment and transform it into a state favorable for the anti-tumor immune response. Secondly, by blocking the interaction between PD-L1 and PD-1, it enhances the ability of T cells to attack tumor cells. Finally, epirubicin exerts a direct cytotoxic effect on tumor cells, further strengthening the elimination of tumor cells by immune cells.

[0094] In short, this drug achieves a synergistic tumor suppression effect that targets the tumor microenvironment by integrating immunotherapy and chemotherapy. This integrated strategy is expected to enhance the effectiveness of tumor treatment and lead to a better quality of life and prognosis for patients.

[0095] (10) The target chemopharmaceutical comprises a DNA carrier, a small molecule compound drug, and an immunostimulant CPG2006, a nucleic acid aptamer CD40, a nucleic acid aptamer PD-L1 in DNA form, and a nucleic acid aptamer C12 linked to the DNA carrier. Here, the DNA carrier further comprises sequences D and E as complementary sequences, where sequence D is SEQ ID NO. 278:5′-GCCACCGTGCTACA-3′ and sequence E is SEQ ID NO. 279:5′-CAGCAGCAGCAGCA-3′. The transition base sequence SEQ ID NO. 280:5′-GTAGCACGGTGGC-3′ is linked to the 3′ end of sequence B in the DNA carrier, sequence D is linked complementaryly to the aforementioned transition base sequence SEQ ID NO. 280:5′-GTAGCACGGTGGC-3′ in a strand-complementary manner, and sequence E is linked complementaryly to sequence C of the DNA carrier in a strand-complementary manner. Preferably, the small molecule compound drug is epirubicin, its free base, or its hydrochloride. Preferably, the DNA carrier includes the tenth sequence group, with a molar ratio of immunostimulant CPG2006 to the DNA carrier being 2:1, and linked to the 5′ end of sequence A and the 5′ end of sequence C, respectively. Preferably, immunostimulant CPG2006 is linked to the 5′ end of sequence A by the base linker "TTTTTT" and to the 5′ end of sequence C by the base linker "TTTTT". The molar ratio of nucleic acid aptamer CD40 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, nucleic acid aptamer CD40 is linked to the 5′ end of sequence B by the base linker "TTTTT". The molar ratio of nucleic acid aptamer PD-L1 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence D. Preferably, the nucleic acid aptamer PD-L1 is linked to the 5′ end of sequence D by the base linker "TTTTTT". The molar ratio of nucleic acid aptamer C12 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence E. Preferably, the nucleic acid aptamer C12 is linked to the 5′ end of sequence E by the base linker "TTTTTT". Specifically, the targeted chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0096] <Drug 13> TIFF2026522667000014.tif90164

[0097] Note: The phosphate bonds between adjacent bases at positions 1-24 of the 5' end of sequences A and C are all phosphorothioate modifications. The phosphate bonds between adjacent bases at positions 1-3 of the 5' end of sequences B, D, and E are all phosphorothioate modifications.

[0098] In the above-mentioned drug 13, the nano-nucleic acid carrier binds to three aptamers (CPG2006, CD40, and PD-L1) and targets the C12 receptor protein, which is highly expressed on the surface of small cell lung cancer cells, thereby enabling the combined targeting of immunotherapy and classical chemotherapy to the tumor microenvironment. This drug can suppress relapsed, refractory, and small cell lung cancer tumor cells with high efficiency.

[0099] CPG2006 aptamers are CpG oligonucleotide mimetic compounds that activate the Toll-like receptor 9 (TLR9) pathway, stimulating the activation and proliferation of dendritic cells and B cells. In this manner, CPG2006 aptamers contribute to the enhancement of the anti-tumor immune response. CD40 aptamers activate dendritic cells, B cells, and other anti-tumor immune cells by binding to the CD40 receptor, thereby promoting the anti-tumor immune response. PD-L1 aptamers block the interaction between PD-L1 and the PD-1 receptor, restoring T cell activity. This contributes to increasing the ability of T cells to attack tumor cells. C12 aptamers specifically target receptor proteins highly expressed on the surface of small cell lung cancer cells. Through the action of C12 aptamers, drugs accurately identify and target tumor cells, increasing the precision of treatment. Epirubicin, a classic chemotherapy drug, blocks DNA replication and transcription, suppressing the proliferation and spread of tumor cells.

[0100] In this integrated nano-nucleic acid carrier, each aptamer and epirubicin act synergistically to enable combined targeting of the tumor microenvironment for immunotherapy and chemotherapy. The CPG2006-CD40-PD-L1 aptamer activates immune cells and enhances the anti-tumor immune response; the C12 aptamer achieves specific targeting of small cell lung cancer cells; and epirubicin exerts a direct cytotoxic effect on tumor cells. This synergistic tumor suppression effect significantly enhances the efficacy of tumor treatment. Specifically, the following points explain the mechanism of action of the integrated nano-nucleic acid carrier in the treatment of small cell lung cancer:

[0101] 1. Improvement of the tumor microenvironment: The CPG2006-CD40-PD-L1 aptamer acts on the tumor microenvironment to reverse the immunosuppressive state. This enhances the anti-tumor immune response and increases the effectiveness of tumor cell elimination. 2. Targeted Delivery: The C12 aptamer specifically binds to a receptor protein highly expressed on the surface of small cell lung cancer cells, enabling precise drug delivery. This targeted delivery strategy reduces distribution in normal tissues and minimizes side effects. 3. Tumor cell apoptosis: Epirubicin inhibits the proliferation and spread of tumor cells by blocking DNA replication and transcription, thereby inducing apoptosis in tumor cells. 4. Restoration of immune surveillance: PD-L1 aptamers block the interaction between PD-L1 and PD-1 receptors, restoring T cell activity. This enhances the immune system's ability to monitor tumor cells, preventing tumor recurrence and metastasis. 5. Formation of immunological memory: CD40 aptamers activate dendritic cells and B cells, contributing to the formation of immunological memory against tumor antigens. This extends the effect of immunotherapy and contributes to preventing tumor recurrence.

[0102] Overall, drug 13 exhibits synergistic tumor-suppressive effects in the treatment of small cell lung cancer. The aptamers and epirubicin work together to improve the tumor microenvironment, enhance the anti-tumor immune response, ensure precise delivery, induce tumor cell apoptosis, restore immune surveillance, and form immunological memory. Such integrated nano-nucleic acid carriers are expected to offer new hope for the treatment of relapsed, refractory, and small cell lung cancer.

[0103] (11) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and an immunostimulant CPG2006 linked to the DNA carrier, nucleic acid aptamer C12, siRNA oligonucleotide CD47, DNA-form nucleic acid aptamer PD-L1, and siRNA oligonucleotide PD-L1, preferably the small molecule compound drug being epirubicin, or its free base or hydrochloride. Preferably the DNA carrier comprises the ninth sequence group, the molar ratio of siRNA oligonucleotide PD-L1 to the DNA carrier is 1:1, the sense strand is linked to the 5′ end of sequence C in the DNA carrier as a transition base sequence, the antisense strand is linked complementaryly to the sense strand as a complementary sequence, and the DNA-form nucleic acid aptamer PD-L1 is linked to the 5′ end of the antisense strand of siRNA oligonucleotide PD-L1. Preferably the nucleic acid aptamer PD-L1 is linked to the 5′ end of the antisense strand of siRNA oligonucleotide PD-L1 by a base linker "TTTTT". The molar ratio of the immunostimulant CPG2006 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably, the immunostimulant CPG2006 is linked to the 5′ end of sequence A by the base linker "TTTTT". The molar ratio of the siRNA oligonucleotide CD47 to the DNA carrier is 1:1, with its sense strand linked to the 5′ end of sequence B as a transition base sequence, and its antisense strand linked complementary to the sense strand as a complementary sequence. The molar ratio of the nucleic acid aptamer C12 to the DNA carrier is 1:1, and it is linked to the 5′ end of the antisense strand of siRNA oligonucleotide CD47. Preferably, the nucleic acid aptamer C12 is linked to the 5′ end of the antisense strand of siRNA oligonucleotide CD47 by the base linker "TTTTT". Specifically, the targeted chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0104] <Drug 14> TIFF2026522667000015.tif95165

[0105] Note: In sequence A, all phosphate bonds between adjacent bases at positions 1-24 of the 5′ end are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 1-3 of the 3′ end are also phosphorothioate modified. In sequence B, all phosphate bonds between adjacent bases at positions 1-3 of both the 5′ and 3′ ends are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 19-21 of the 5′ end of sequence B are phosphorothioate modified. In sequence C, all phosphate bonds between adjacent bases at positions 1-3 of both the 5′ and 3′ ends are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 19-21 of the 5′ end of sequence C are phosphorothioate modified. In sequence D, all phosphate bonds between adjacent bases at positions 1-3 of both the 5′ and 3′ ends are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 19-21 of the 3′ end of sequence D are phosphorothioate modified. All phosphate bonds between adjacent bases at positions 1-3 at both the 5′ and 3′ ends of sequence E are phosphorothioate modifications, and all phosphate bonds between adjacent bases at positions 19-21 at the 3′ end of sequence E are phosphorothioate modifications. The underlined RNA sequence shows C / U 2′-F modification.

[0106] In the above drug 14, the nanonucleic acid carrier is modified by a sequence extension method, and CpG2006 and C12 / PD-L1 aptamers are used as active targeting factors to specifically bind to HDLBP (high-density lipoprotein-binding protein) and PD-L1 protein, which are highly expressed on the surface of cancer cells. At the same time, gene regulatory factors CD47 siRNA and PD-L1 siRNA are incorporated, and approximately 30 GC / CG sites for epirubicin loading are designed on three double-stranded DNA arms formed by the self-assembly of three core skeletal sequences. In the manufacturing process, it is preferably used in an excess ratio of 40:1 (epirubicin:nanonucleic acid carrier), and 4% paraformaldehyde is used in combination as a coupling agent. The anthracycline tetracyclic skeleton of the epirubicin molecule specifically intercalates at the GC / CG sites of the nanonucleic acid carrier. The amino group of its daunosamine sugar and the guanosine outer ring amino group at the GC / CG binding site on the carrier side form a covalent bond reaction of Schiff bases under paraformaldehyde catalyst, thereby constructing a stable support mode consisting of "physical intercalation + covalent bonding." These nanoparticles possess a nanoparticle-specific passive targeting "funnel" accumulation effect on tumor tissue, and also exhibit target affinity for tumor tissues and cells with high expression of HDLBP protein and PD-L1 protein. These two targeting effects mediate high concentration accumulation at tumor sites. After the C12 aptamer binds to the HDLBP protein highly expressed on the surface of tumor cells, it is taken up into the cytoplasm by mechanisms such as endocytosis. Under reducing conditions such as low pH and high glutathione concentrations within tumor cells, epirubicin is dissociated and released from the nanocarrier. The released epirubicin then translocates to the nucleus and specifically binds and intercalates at the GC / CG positions of nuclear double-stranded DNA, thereby inhibiting DNA transcription and suppressing the differentiation and proliferation of tumor cells, thus exerting an antitumor effect that suppresses tumor development and progression.

[0107] The above drug 14 combines immunotherapy, gene knockdown, and chemotherapy to comprehensively suppress advanced, recurrent, metastatic, and refractory small cell lung cancer. The mechanisms of action of each component are as follows:

[0108] 1. CpG 2006: CpG 2006 is a CpG oligonucleotide that activates Toll-like receptor 9 (TLR9) and modulates the immune response. TLR9 activation enhances the antitumor immune response through activation of dendritic cells and B cells, promotion of Th1 cytokine production, and enhancement of cytotoxic T cells and NK cells. 2. C12: C12 is an aptamer that targets HDLBP (high-density lipoprotein-binding protein), which is highly expressed on the surface of small cell lung cancer cells. This allows nanoparticles to be specifically guided to tumor cells, enhancing therapeutic efficacy and reducing toxicity to normal tissues. 3. CD47 siRNA: CD47 siRNA is a small interfering RNA that knocks down the expression of the CD47 protein, suppressing immune evasion by tumor cells. CD47 generally functions as a "don't eat me" signal and binds to SIRPα to suppress phagocytosis of tumor cells by macrophages. Decreasing CD47 expression restores the phagocytic function of macrophages, enhancing the anti-tumor immune effect. 4. PD-L1: PD-L1 aptamers are aptamers that target PD-L1. They bind to PD-L1, blocking its interaction with the PD-1 receptor and activating the anti-tumor immune response. The PD-1 / PD-L1 pathway is crucial for tumor immune evasion, and blocking it increases the aggressiveness of immune cells against tumor cells. 5. PD-L1 siRNA: PD-L1 siRNA targets the PD-L1 gene, reducing the expression of the PD-L1 protein and enhancing the immune cells' attack on tumor cells. When combined with the Apt PD-L1 aptamer, it further enhances the efficiency of blocking the PD-1 / PD-L1 pathway and activates the immune response. 6. The nano-nucleic acid carrier is a nano-nucleic acid carrier formed by a self-assembly mechanism in which three partially continuous sequences complementarily pair with each other. By extending the sequences, multiple effective nucleic acid therapeutic molecules are linked together, enhancing the stability and targeting of the therapeutic molecules and realizing synergistic therapy. 7. Epirubicin, as a DNA topoisomerase II inhibitor, inhibits DNA replication and transcription processes, thereby suppressing tumor cell proliferation. By delivering epirubicin specifically to tumor tissue and tumor cells using nanoparticles, and then releasing it under low pH and high glutathione-reducing conditions, the concentration within the tumor tissue can be increased, reducing systemic toxicity.

[0109] In general, the mechanisms of action of the above-mentioned drug 14 include activation of the immune response by CpG 2006, tumor targeting by C12 aptamer, dual blockade of immune evasion by CD47 siRNA and PD-L1 siRNA, and inhibition of tumor cell proliferation by epirubicin. Such nanoparticle therapy integrates multifaceted means to simultaneously suppress advanced, recurrent, metastatic, and refractory cancers (e.g., small cell lung cancer) through multiple pathways.

[0110] (12) The target chemical drug comprises a DNA carrier, a small molecule compound drug, and miRNA oligonucleotide A-miR-21, siRNA oligonucleotide TGF-β1, nucleic acid aptamer TfRA4, and nucleic acid aptamer PD-L1 linked to the DNA carrier, preferably epirubicin, its free base, or its hydrochloride as the small molecule compound drug. Preferably, the DNA carrier comprises the second sequence group, with a molar ratio of siRNA oligonucleotide TGF-β1 to the DNA carrier of 1:1, where its antisense strand is linked to the 3′ end of sequence C as a transition base sequence, and its sense strand is linked complementary to the antisense strand as a complementary sequence. The molar ratio of nucleic acid aptamer TfRA4 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence A. Preferably, nucleic acid aptamer TfRA4 is linked to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of nucleic acid aptamer PD-L1 to the DNA carrier is 1:1, and it is linked to the 5′ end of sequence B. Preferably, the nucleic acid aptamer PD-L1 is linked to the 5′ end of sequence B by the base linker "TTTTT". The molar ratio of miRNA oligonucleotide A-miR-21 to the DNA carrier is 2:1, and they are sequentially linked to the 5′ end of the sense strand of siRNA oligonucleotide TGF-B. Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by the self-assembly of the following sequences, and epirubicin supported on the targeted nucleic acid carrier:

[0111] <Drug 15> TIFF2026522667000016.tif75165

[0112] Note: In sequence B, all phosphate bonds between adjacent bases at positions 1-3 of the 5′ end are phosphorothioate modified. In sequence C, all phosphate bonds between adjacent bases at positions 29-31 of the 5′ end are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 1-3 of the 3′ end are also phosphorothioate modified. In sequence D, all phosphate bonds between adjacent bases at positions 1-3 of the 3′ end are phosphorothioate modified, and all phosphate bonds between adjacent bases at positions 19-21 are also phosphorothioate modified. C / U 2′-F modification in the underlined RNA sequence.

[0113] The above drug 15 is a nano-nucleic acid integrated aptamer active gene combination immunotherapy agent that enables combined targeting of the tumor microenvironment with immunotherapy and classical chemotherapy, and can efficiently suppress tumor cells of recurrent or resistant glial tumors or metastatic brain tumors. The functions of each effector molecule and their synergistic effects in tumor suppression are shown below:

[0114] 1. A-miR-21: A-miR-21 is an ASO that targets microRNA-21 (miR-21). miR-21 is a widely expressed microRNA and is involved in the development, progression, invasion, and metastasis of various tumors. A-miR-21 binds to miR-21 and suppresses the regulation of its target genes, thereby reducing the proliferation, invasion, and metastatic capacity of tumor cells. 2. TGF-β1 siRNA: TGF-β1 siRNA is an siRNA that specifically knocks down TGF-β1. TGF-β1 is overexpressed in the tumor microenvironment and promotes the growth, invasion, and metastasis of tumor cells. TGF-β1 siRNA reduces TGF-β1 expression and suppresses the growth and metastasis of tumor cells. 3. Apt TfRA4: TfRA4 is a nucleic acid aptamer that targets the transferrin receptor (TfR). TfR is highly expressed in tumor cells and the blood-brain barrier (BBB), and Apt TfRA4 binds to TfR, enabling active targeting of tumor cells and the BBB. This increases the drug concentration in tumor tissue and reduces toxicity to normal tissue. 4. Apt PD-L1: The PD-L1 aptamer is a nucleic acid aptamer that targets programmed cell death ligand 1 (PD-L1) on the surface of tumor cells. PD-L1 binds to PD-1 on immune cells, suppressing immune cell activity and causing tumor cells to evade immune surveillance. Apt PD-L1 binds to PD-L1, blocking its interaction with PD-1, restoring immune cell activity, and enhancing the effect of tumor immunotherapy. 5. The nano-nucleic acid carrier skeleton in the pharmaceutical product integrates and binds molecules such as A-miR-21, TGF-β1 siRNA, TfRA4 aptamer, and PD-L1 aptamer to constitute the target drug. The nano-nucleic acid carrier skeleton stabilizes the overall structure and improves the biostability and duration of action of the drug. 6. Epirubicin: Epirubicin is a widely used antitumor drug that primarily inhibits tumor growth by suppressing DNA synthesis and cell division in tumor cells. By covalently loading epirubicin onto a nanonucleic acid carrier scaffold, combination therapy with immunotherapeutic agents is realized, enhancing the antitumor effect.

[0115] Specifically, the above drug 15 achieves highly efficient treatment through the following synergistic tumor-suppressing effects: 1. Combination Targeting: A-miR-21, TGF-β1 siRNA, TfRA4 aptamer, and PD-L1 aptamer target miR-21, TGF-β1, TfR, and PD-L1 on tumor cells, respectively, suppressing tumors through multiple pathways. Such combination targeting strategies contribute to enhancing drug efficacy and reducing side effects. 2. Combination therapy: By combining epirubicin, a classic chemotherapy drug, with gene therapy agents such as A-miR-21 and TGF-β1 siRNA, and PD-L1 aptamer immunotherapy agents, comprehensive treatment is achieved, enhancing the antitumor effect. 3. Synergistic immunotherapy: PD-L1 aptamers release immunosuppression and activate immune cells in the tumor microenvironment, exerting a synergistic antitumor effect in combination with chemotherapy drugs such as epirubicin. 4. Knockdown of key signaling pathways: A-miR-21 and TGF-β1 siRNA knock down key signaling pathways within tumor cells, suppressing tumor cell growth, invasion, and metastasis, thereby enhancing the antitumor effect of drugs.

[0116] Overall, drug 15 demonstrates synergistic effects in tumor suppression by utilizing diverse targeting strategies and combination therapy approaches. Specific findings of synergistic tumor suppression are presented below: 5. Improved drug penetration: The TfRA4 aptamer binds to TfR, making it easier for the entire nanopharmaceutical to cross the blood-brain barrier (BBB), increasing the drug concentration in tumor tissue. This is important for the treatment of cerebral colloid tumors and brain metastatic tumors. 6. Reduction of toxicity and side effects: Multi-pathway targeting and combination therapy strategies allow for reduced dosage, thereby decreasing toxicity to normal cells and tissues. Furthermore, integrating the drug into nano-nucleic acid carriers extends biostability and duration of action, further reducing side effects. 7. Countering drug resistance: Combination therapy strategies can overcome drug resistance that may arise with monotherapy. By simultaneously targeting multiple signaling pathways and mechanisms in tumor cells, drug resistance in tumor cells is reduced and drug efficacy is enhanced. 8. Extended Survival Time and Improved Quality of Life: Drug 15 can effectively control tumor growth, invasion, and metastasis through multi-pathway synergistic inhibition, potentially extending patient survival time. Simultaneously, the reduction of toxicity and side effects contributes to an improved quality of life (QOL) for patients.

[0117] In short, the above drug 15, as a nano-nucleic acid-integrated aptamer active gene combination immunotherapy agent, allows each effector molecule to exert a synergistic tumor suppressive effect, enabling targeting of the tumor microenvironment through the combined use of immunotherapy and classical chemotherapy. This innovative treatment has significant value in efficiently suppressing tumor cells in recurrent or resistant cerebrovascular tumors or metastatic brain tumors.

[0118] (13) The target chemotherapy comprises a DNA carrier, a small molecule chemotherapy agent, and nucleic acid aptamers IL-4Rα, siRNA oligonucleotide CD47, DNA-type nucleic acid aptamer TfRA4, siRNA oligonucleotide PD-L1, and nucleic acid aptamer GPC-1 bound to the DNA carrier. Preferably, the small molecule chemotherapy agent is doxorubicin (free base or hydrochloride). The DNA carrier comprises a group 3 sequence, with a molar ratio of nucleic acid aptamer TfRA4 to DNA carrier of 1:1, and bound to the 5′ end of sequence A. Preferably, nucleic acid aptamer TfRA4 is bound to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of nucleic acid aptamer GPC-1 to DNA carrier is 1:1, and bound to the 5′ end of sequence C. The molar ratio of siRNA oligonucleotide PD-L1 to the DNA carrier is 1:1, with its sense strand linked to the 5′ end of sequence B as an intermediate base segment (bridge), and its antisense strand linked complementaryly to the sense strand as a complementary sequence, and also linked to the 3′ end of sequence A. The molar ratio of siRNA oligonucleotide CD47 to the DNA carrier is 1:1, with its sense strand linked to the 3′ end of sequence C as an intermediate base segment, and its antisense strand linked complementaryly to the sense strand as a complementary sequence. The molar ratio of nucleic acid aptamer IL-4Rα to the DNA carrier is 1:1, and it is bound to the 5′ end of the antisense strand of siRNA oligonucleotide CD47. Preferably, the nucleic acid aptamer IL-4Rα is bound to the 5′ end of the antisense strand of siRNA oligonucleotide CD47 by the base linker "AAAA". Specifically, the targeted chemotherapy consists of a targeted nucleic acid carrier that self-assembles according to the following sequence, and doxorubicin loaded onto the targeted nucleic acid carrier:

[0119] <Drug 16> TIFF2026522667000017.tif81166

[0120] Note: Each adjacent bond between bases 1-3 at the 3′ end of sequence A, and between bases 19-21 at the 3′ end of sequence A, is modified with phosphorothioate (PS). Each adjacent bond between bases 1-3 at the 5′ end of sequence B, and between bases 19-21 at the 5′ end of sequence B, is modified with PS. Each adjacent bond between bases 1-3 at the 3′ end of sequence C, and between bases 19-21 at the 3′ end of sequence C, is modified with PS. The area between bases 1-4 at the 5′ end of sequence D is modified with PS, and the area between bases 1-3 at the 3′ end of sequence D, and the area between bases 19-21 at the 3′ end of sequence D are also modified with PS. In the underlined RNA sequences in sequences A, B, and C, 2′-fluoro(2′F) modification is applied to C / U. In sequence D, the first 44 bases undergo a 2′F modification to the C / U bond and a 2′-O-methyl (2′-OMe) modification to the A / G bond, while the 45-64 bases undergo a 2′F modification to the C / U bond.

[0121] The above-mentioned drug 16, as a nano-nucleic acid integrated aptamer active gene combination immunotherapy agent, combines multiple therapeutic strategies such as immunotherapy, targeted therapy, gene therapy, and classical chemotherapy to achieve combined targeting of the tumor microenvironment. This integrated therapy scheme shows extremely high efficacy in suppressing tumor cells in recurrent / resistant tumors, glioblastoma, or brain metastatic tumors. The function of each effector molecule and its role in synergistic antitumor action are shown below:

[0122] 1. IL-4Rα: A subunit of the interleukin-4 receptor, it modulates the polarization of paratumor macrophages (TAMs) as an immunotherapy target, influencing the tumor microenvironment. Controlling TAM polarization can suppress tumor growth and metastasis. 2. CD47 siRNA: CD47 is an immune checkpoint protein that inhibits macrophage phagocytosis of tumor cells through a "don't eat me" signal. CD47 siRNA specifically knocks down CD47 expression, deactivating this signal and enhancing macrophage phagocytosis of tumor cells. 3. TfRA4 aptamer: Specifically binds to transferrin receptors on the blood-brain barrier (BBB), promoting the passage of nanodrugs across the BBB, enhancing delivery to brain tumor tissue, and increasing drug concentration at the tumor site. 4. PD-L1 siRNA: PD-L1 is an immune checkpoint protein that binds to PD-1, suppressing T cell activity and reducing the immune system's ability to clear tumors. PD-L1 siRNA specifically knocks down PD-L1 expression, releasing its suppression of T cells and enhancing immune clearing ability. 5. GPC-1 aptamer: Specifically binds to and inhibits glioblastoma or brain metastatic tumor cells that highly express glypican-1 (GPC-1), exerting selective suppression against tumor cells and inhibiting tumor growth and metastasis. 6. Nanonucleic acid carriers: These carriers enable the combination and deployment of various aptamers and siRNAs, realizing highly integrated nanopharmaceutical designs that improve drug targeting and biological activity. 7. Doxorubicin: A classic antitumor drug that inhibits DNA replication and cell division in tumor cells. By combining doxorubicin with other immunotherapy and gene therapies using nanocarriers, the benefits of combination therapy are realized, and the antitumor effect is enhanced.

[0123] Through the synergistic action of the aforementioned effector molecules, this drug, based on a nano-nucleic acid-integrated aptamer active gene combination immunotherapy agent, demonstrates remarkable superiority in suppressing tumor cells in relapsed / resistant glioblastoma or brain metastatic tumors. This combination therapy strategy simultaneously achieves highly efficient suppression of these tumor cells through pathways such as regulation of the tumor microenvironment, enhancement of the immune system's tumor scavenging ability, promotion of blood-brain barrier crossing, and specific suppression of tumor cells.

[0124] (14) The target chemoagent comprises a DNA carrier, a small molecule chemoagent, and DNA-type nucleic acid aptamers AS1411, EGFR, and A15 bound to the DNA carrier, with gemcitabine being the preferred small molecule chemoagent. Preferably, the DNA carrier comprises a group 4 sequence, with a molar ratio of 1:1 between DNA-type nucleic acid aptamer AS1411 and the DNA carrier, and bound to the 5′ end of sequence A. The molar ratio of EGFR to the DNA carrier is 1:1, and bound to the 5′ end of sequence B. The molar ratio of A15 to the DNA carrier is 1:1, and bound to the 5′ end of sequence C. Preferably, gemcitabine is linked in the form of at least one base insertion at one or more of the following positions: between DNA nucleic acid aptamer AS1411 and sequence A, at the 3′ end of sequence A; between DNA nucleic acid aptamer EGFR and sequence B, at the 3′ end of sequence B; between DNA nucleic acid aptamer A15 and sequence C, at the 3′ end of sequence C. Specifically, the target chemotherapy is a gemcitabine-targeting drug formed by self-assembly of the following sequences:

[0125] <Drug 17> TIFF2026522667000018.tif44166

[0126] Note: The above J represents gemcitabine, which is synthesized directly into the sequence during sequence synthesis in the form of base substitutions and base extensions. This corresponds to 36 gemcitabines bound to a single DNA target carrier.

[0127] SEQ ID NO. 103: 5'-GGTGGTGGTGGTTGTGGTGGTGGTGG-3'; SEQ ID NO. 15: 5'-GCGGCGCCACCGAGCGTTCCGGGAGAGGCC-3'; SEQ ID NO. 139: 5'-GCCTTAGTAACGTGCTTTGATGTCGATTCGACAGGAGGC-3'; SEQ ID NO. 16: 5'-GGCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; SEQ ID NO. 102: 5'-CCCTCCTACATAGGG-3'; SEQ ID NO. 14: 5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'.

[0128] In drug 17 described above, highly specific targeting of tumor cells was achieved by integrating different aptamers using a nano-nucleic acid carrier. Here, gemcitabine can be directly incorporated into the carrier aptamer sequence using an oligonucleotide synthesizer during synthesis. The functions of each aptamer are as follows:

[0129] 1. AS1411: A nucleoline aptamer that specifically binds to nucleoline receptors on the surface of tumor cells, thereby achieving targeting of tumor cells. 2. EGFR: An aptamer that has the ability to specifically bind to the epidermal growth factor receptor (EGFR). Since EGFR is overexpressed in many tumor cells, this aptamer enhances the targeting of nano-nucleic acid carriers against tumor cells. 3. A15: An aptamer that specifically binds to the CD133 receptor protein, a stem cell-like marker. This allows for more precise targeting of tumor stem cells, potentially combating tumor recurrence and drug resistance. 4. Nanonucleic acid carriers: These function as a basic framework, integrating each aptamer with the drug and improving the bioactivity and targeting of the drug. 5. Gemcitabine: A widely used antitumor drug that inhibits DNA synthesis and cell division. By binding gemcitabine to other aptamers using nano-nucleic acid carriers, highly specific targeting of tumor cells is achieved.

[0130] In general, the above-mentioned drug 17 achieves highly specific targeting of tumor cells by combining multiple aptamers. By targeting nucleolin, EGFR, and CD133 receptors, the nano-nucleic acid carrier exerts effective inhibitory effects on tumor cells, particularly tumor stem cells. The introduction of gemcitabine further enhances the antitumor effect, and this nano-nucleic acid carrier has good potential for clinical application.

[0131] (15) The target chemoagent comprises an RNA carrier, a small molecule chemoagent, and DNA-type nucleic acid aptamers AS1411, EGFR, and A15 bound to the RNA carrier, with 5-fluorouracil (5-FU) being the preferred small molecule chemoagent. Preferably, the RNA carrier comprises a group 5 sequence, with a molar ratio of 1:1 between DNA-type nucleic acid aptamer AS1411 and the RNA carrier, and bound to the 5′ end of sequence A. The molar ratio of EGFR to the RNA carrier is 1:1, and bound to the 5′ end of sequence B. The molar ratio of A15 to the RNA carrier is 1:1, and bound to the 5′ end of sequence C. Preferably, 5-fluorouracil is linked in the form of at least one base insertion at one or more of the following positions: between DNA nucleic acid aptamer AS1411 and sequence A, at the 3′ end of sequence A; between DNA nucleic acid aptamer EGFR and sequence B, at the 3′ end of sequence B; between DNA nucleic acid aptamer A15 and sequence C, at the 3′ end of sequence C. Specifically, the target chemologic agent is a 5′-fluorouracil targeting agent formed by self-assembly of the following sequences:

[0132] <Drug 18> TIFF2026522667000019.tif62164

[0133] Note: Above, N represents 5′-fluorouracil (5-FU), which is directly synthesized into the sequence in the form of base substitution and base extension during sequence synthesis; any bond between any two adjacent 5′-fluorouracil bases is considered a PS modification; this corresponds to 36 5′-fluorouracils being bound to a single DNA target carrier. The two bases before the 5′ end of sequence A are PS modified; the 1-2 bases at the 5′ and 3′ ends of sequence B are PS modified; the 1-2 bases at the 5′ end of sequence C are PS modified. For RNA carriers, C / U is modified with 2′-fluoro(2′F) and A / G with 2′-O-methyl(2′-OMe), and the 1-2 bases at the 5′ and 3′ ends are PS modified. In the underlined RNA sequence portions, C / U is modified with 2′F and A / G with 2′-OMe.

[0134] In drug 18 described above, multiple aptamers were integrated using a nanonucleic acid carrier, achieving highly specific targeting of tumor cells. Here, 5′-fluorouracil nucleoside can be directly incorporated into the carrier aptamer sequence using an oligonucleotide synthesizer during synthesis. The functions of each component of the nanonucleic acid carrier are as follows:

[0135] 1,5′-Fluorouracil: A widely used antitumor drug that inhibits DNA and RNA synthesis. 5′-Fluorouracil nucleoside is a nucleoside analog of 5-FU and may be more efficiently taken up into cells and exert its therapeutic effect. 2. AS1411: A nucleoline aptamer that specifically binds to nucleoline receptors on the surface of tumor cells, thereby achieving targeting of tumor cells. 3. EGFR: An aptamer that has the ability to specifically bind to the epidermal growth factor receptor (EGFR). Since EGFR is overexpressed in many tumor cells, this aptamer enhances the targeting ability of nanonucleic acid carriers against tumor cells. 4. A15: An aptamer that specifically binds to the CD133 receptor protein, a stem cell-like marker. This allows for precise targeting of tumor stem cells, potentially combating tumor recurrence and drug resistance. 5. Nanonucleic acid carriers: These function as a basic framework, integrating each aptamer with the drug to improve the drug's biological activity and targeting ability.

[0136] Based on the above, drug 18 achieves highly specific targeting of tumor cells by integrating multiple aptamers. This nano-nucleic acid carrier exerts effective inhibitory effects on tumor cells, particularly tumor stem cells, by utilizing strategies that target nucleolin, EGFR, and CD133 receptors. Simultaneously, the introduction of 5′-fluorouracil nucleoside further enhances the antitumor effect, giving this nano-nucleic acid carrier excellent potential for clinical application.

[0137] (16) The target chemologic drug comprises a DNA carrier, a small molecule chemologic drug, and DNA-type nucleic acid aptamers AS1411, EGFR, and A15 bound to the DNA carrier, with paclitaxel being the preferred small molecule chemologic drug. Preferably, the DNA carrier comprises a group 6 sequence, with a molar ratio of 1:1 between Preferably, the nucleic acid aptamer A15 is bound to the 5′ end of sequence C by the base linker "TTTTT". Specifically, the targeted chemotherapy comprises a targeted nucleic acid carrier formed by self-assembly via the following sequence, and paclitaxel mounted thereon:

[0138] <Drug 19> TIFF2026522667000020.tif61164

[0139] Note: The 5′ and 3′ ends of sequences A, B, and C above are all modified with phosphorothioate (PS) between the 1st and 2nd bases. In sequence A above, the 35th, 39th, 54th, and 59th bases from the 5′ end are modified with amino(-NH2) molecules. In sequence B above, the 50th, 55th, 66th, and 70th bases from the 5′ end are modified with amino(-NH2) molecules. In sequence C above, the 24th and 28th bases from the 5′ end are modified with amino(-NH2) molecules.

[0140] In drug 19 described above, the nanonucleic acid carrier simultaneously synthesizes multiple aminolinkers at the designed site during the synthesis of three DNA oligonucleotide / aptamer base sequences. These linkers can then be covalently linked to linker fragments having disulfide bonds modified on the 2′-OH of paclitaxel via the action of a coupling agent. Each nanoparticle carries 8 to 12 paclitaxel molecules and, under the mediation of tri-aptamer complex targeting, efficiently accumulates in the tumor microenvironment, is taken up by target cells, and releases paclitaxel molecules. These targeted paclitaxel nanoparticles can efficiently suppress various paclitaxel-sensitive tumors, possessing many advantages such as high targetability, controlled release, and low toxicity, thus improving the efficacy of paclitaxel therapy and reducing side effects. Specifically, this drug has the following properties:

[0141] 1. High Targetability: The combination of AS1411, EGFR, and A15 aptamers gives the nanoparticles high targetability. The AS1411 aptamer primarily targets nucleolins in tumor cells, the EGFR aptamer targets epidermal growth factor receptors on tumor cells, and the A15 aptamer selectively recognizes and binds to tumor stem cell-like proteins. The synergistic effect of these three aptamers allows the nanoparticles to efficiently accumulate in the tumor microenvironment and penetrate target cells. 2. Controlled Release: Paclitaxel molecules are covalently bonded to nanoparticles via disulfide linkers. When the nanoparticles enter tumor cells, the disulfide bonds are cleaved by the intracellular reducing environment (e.g., high concentrations of glutathione), and the paclitaxel molecules are released. This controlled release mechanism reduces nonspecific accumulation in normal tissues and contributes to reduced toxicity. 3. Low toxicity: Due to its highly targeted and controlled-release properties, paclitaxel's nonspecific accumulation in normal tissues is reduced, mitigating its toxicity to normal tissues. This is advantageous for improving therapeutic efficacy and reducing side effects. 4. Antitumor effect: The released paclitaxel molecules disrupt the balance of microtubule polymerization and depolymerization dynamics, suppressing mitosis in tumor cells and inducing apoptosis. Furthermore, it suppresses tumor progression and metastasis through pathways such as the inhibition of cell migration, invasion, and angiogenesis. 5. Addressing drug resistance: By simultaneously targeting multiple targets with three aptamers, the possibility of paclitaxel resistance development in tumor cells is reduced. Furthermore, high targeting and controlled release maintain high paclitaxel concentrations in the tumor microenvironment, further enhancing the killing effect against resistant tumor cells. 6. Combination Therapy: The AS1411-EGFR-A15-DNA-paclitaxel nanonucleic acid carrier may exert synergistic tumor-suppressing effects when used in combination with other antitumor drugs, radiotherapy, immunotherapy, etc. For example, when used in combination with immunotherapy, it may enhance the antitumor activity of immune cells by improving the tumor microenvironment, and when used in combination with radiotherapy, it may increase the radiosensitivity of tumor cells through its radiosensitizing effect. 7. Personalized therapy: Personalized therapy can be achieved by adjusting the combination of aptamers according to the expression status of tumor cell surface targets for each patient and custom designing nano-nucleic acid carriers.

[0142] Overall, the above-mentioned drug 19 exhibits excellent antitumor activity due to its characteristics such as high targeting ability, controlled release, and low toxicity. Through combination therapy with other treatments, management of drug tolerance, and realization of personalized medicine, this nano-nucleic acid carrier is expected to become an effective treatment method for a variety of tumors.

[0143] (17) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and DNA-type nucleic acid aptamers AS1411, EGFR, and A15 bound to the DNA carrier, with meitansine being the preferred small molecule chemical agent. Preferably, the DNA carrier comprises a group 7 sequence, the molar ratio of DNA-type nucleic acid aptamer AS1411 to the DNA carrier is 1:1, and it is bound to the 5′ end of sequence A. Preferably, nucleic acid aptamer AS1411 is bound to the 5′ end of sequence A by a base linker "TTTTT". The molar ratio of DNA-type nucleic acid aptamer EGFR to the DNA carrier is 1:1, and it is bound to the 5′ end of sequence B. Preferably, nucleic acid aptamer EGFR is bound to the 5′ end of sequence B by a base linker "TTTTT". The molar ratio of DNA-type nucleic acid aptamer A15 to the DNA carrier is 1:1, and it is bound to the 5′ end of sequence C. Preferably, the nucleic acid aptamer A15 is bound to the 5′ end of sequence C by the base linker "TTTTT". Specifically, the target chemologic drug comprises a targeted nucleic acid carrier formed by self-assembly according to the following sequence, and maytansine mounted thereon:

[0144] <Drug 20> TIFF2026522667000021.tif61165

[0145] Note: The 5′ and 3′ ends of sequences A, B, and C above are all modified with phosphorothioate (PS) between the 1st and 2nd bases. In sequence A, the 43rd and 61st bases from the 5′ end are modified with amino(-NH2); in sequence B, the 48th and 63rd bases from the 5′ end are modified with amino(-NH2); and in sequence C, the 24th and 43rd bases from the 5′ end are modified with amino(-NH2).

[0146] In drug 20 described above, the nanonucleic acid carrier simultaneously synthesizes multiple aminolinkers at the designed site during the synthesis of three DNA oligonucleotide / aptamer base sequences. These linkers can then be covalently linked to the linker fragment modified on the 2′-OH of maytansine via a coupling reaction. The nanoparticles carry six maytansine molecules and, under the mediation of tri-aptamer complex targeting, efficiently accumulate in the tumor microenvironment, are taken up by target cells, and release highly toxic maytansine molecules. Such targeted paclitaxel nanoparticles can efficiently suppress various maytansine-sensitive tumors.

[0147] The above-mentioned drug accumulates efficiently in the tumor microenvironment under the mediated by tri-aptamer complex targeting, is taken up by target cells, and releases highly toxic maytansine molecules. These targeted nanoparticles can efficiently suppress various maytansine-sensitive tumors. The mechanism of action of these nanoparticles is analyzed in detail below:

[0148] 1. Targetability: The AS1411-EGFR-A15-DNA-Maytansin nanonucleic acid carrier achieves high-level targeting of tumor cells through three aptamers: AS1411, EGFR, and A15. AS1411 specifically binds to the nucleolin receptor, the EGFR aptamer specifically binds to the epidermal growth factor receptor, and the A15 aptamer binds to the CD133 receptor protein. These three aptamers allow the nanonucleic acid carrier to accurately identify and target tumor cells. 2. Drug Loading Capacity: In the design, the meitansine molecule is covalently bonded to the DNAh oligonucleotide / aptamer base sequence via an aminolinker, allowing each nanoparticle to carry six meitansine molecules, thereby improving the amount of drug loaded. 3. Drug Release: After the nano-nucleic acid carrier is taken up by tumor cells, under appropriate conditions, the maytansine molecule is released from the linker site, exerting a toxic effect on tumor cells. Maytansine is a DNA synthesis inhibitor, blocking DNA synthesis in tumor cells and inducing apoptosis. 4. Tumor suppression effect: It efficiently suppresses tumor cells through its targeting, loading capacity, and controlled release properties. Furthermore, it suppresses tumor progression and metastasis through pathways such as inhibition of cell migration, invasion, and angiogenesis. 5. Addressing drug resistance: By targeting multiple targets simultaneously with three aptamers, the possibility of developing resistance to meitansine is reduced. At the same time, high targeting and controlled release reduce toxicity to normal cells, improving therapeutic efficacy and patient tolerability. 6. In vivo distribution and pharmacokinetics: The design of nano-nucleic acid carriers provides advantageous in vivo distribution for tumor treatment. They exhibit high stability in the circulatory system and can effectively avoid rapid clearance by non-target organs such as the liver and kidneys. Furthermore, nano-nucleic acid carriers achieve high accumulation in tumor tissue through the EPR effect (Enhanced Permeability and Retention). 7. Safety and Biocompatibility: The materials and synthesis processes exhibit good biocompatibility, reducing the likelihood of adverse events during treatment. Furthermore, the high targeting and controlled release properties reduce toxicity to normal cells, improving safety.

[0149] In general, the above-mentioned drug 19 accumulates efficiently in the tumor microenvironment under the mediation of tri-aptamer complex targeting, is taken up by target cells, and releases highly toxic maytansine molecules. By utilizing the advantages of targetability, drug loading capacity, controlled release, tumor suppression, drug resistance management, in vivo distribution and pharmacokinetics, safety and biocompatibility, these targeted nanoparticles can efficiently suppress various maytansine-sensitive tumors.

[0150] (18) The targeted chemologic drug comprises a DNA carrier, a small molecule chemologic drug, and nucleic acid aptamers MUC1, AS1411, and GPC-1 bound to the DNA carrier, with gemcitabine being the preferred small molecule chemologic drug. Preferably, the DNA carrier comprises a group 9 sequence, with a molar ratio of nucleic acid aptamer MUC1 to the DNA carrier being 1:1 and bound to the 5′ end of sequence A. The molar ratio of nucleic acid aptamer AS1411 to the DNA carrier being 1:1 and bound to the 5′ end of sequence B. The molar ratio of nucleic acid aptamer GPC-1 to the DNA carrier being 1:1 and bound to the 5′ end of sequence C. Specifically, the targeted chemologic drug comprises a targeted nucleic acid carrier formed by self-assembly of the following sequences, and doxorubicin mounted thereon:

[0151] <Drug 21> TIFF2026522667000022.tif44165

[0152] Note: The above J represents gemcitabine, which is directly synthesized into the sequence in the form of base substitution and base extension during sequence synthesis; all adjacent bases between gemcitabines are phosphorothioate (PS) modified; this corresponds to 36 gemcitabines bound to a single DNA target carrier. The bases between positions 1 and 2 at the 5′ end of sequences A, B, and C above are all PS modified.

[0153] SEQ ID NO. 160:5'-GCAGTTGATCCTTTGGATACCCTGG-3'; SEQ ID NO. 26:5'-GCGACGCCCACGAGCGTTCCGGGAGAGGAGC-3'; SEQ ID NO. 103:5'-GGTGGTGGTGGTTGTGGTGGTGGTGG-3'; SEQ ID NO. 11:5'-GCTCCTCTCCCGGTTCGCGCGAGCCGCG-3'; SEQ ID NO. 145:5'-AACGGAGTGTGGCTAACTCGA-3'; SEQ ID NO. 27:5'-CGCGGCTCGCGGCCATAGCCGTGGGCGTCGC-3'.

[0154] In drug 21 described above, the nanonucleic acid carrier is formed by synthesizing gemcitabine molecules, which are arranged in sequence with three aptamer sequences, using an oligonucleotide synthesizer. These three DNA carrier core oligonucleotides are then combined according to the designed base sequence using a sequence extension method, and the resulting molecules are directly synthesized into each sequence (there is a complementary sequence relationship between the parts of the three single strands). Subsequently, these are combined based on the principle of base complementarity self-assembly to form the MUC1 / AS1411 / GPC1-gemcitabine-DNAh carrier. Next, doxorubicin is covalently modified to the pre-designed GC / CG mounting sites (30 locations) of the nanonucleic acid carrier by physical intercalation and amino crosslinking, forming the MUC1 / AS1411 / GPC1-gemcitabine-DNAh-doxorubicin targeted nanoparticle drug. Due to its complex targeting mediation function, the nanoparticles specifically bind to and accumulate in the tumor microenvironment and tumor cells that highly express the MUC1 / nucleolin / GPC1 protein. Doxorubicin is reductively dissociated and released under the low pH and high glutathione environment within tumor cells. The dissociated doxorubicin and gemcitabine, dissociated by enzymatic cleavage, specifically bind to the "GC / CG" target of double-stranded DNA chromosomes in the nucleus, inhibiting DNA transcription. Furthermore, gemcitabine inhibits DNA synthesis as an alternative, thereby preventing tumor cell division and proliferation and suppressing tumor development and progression. The MUC1 / AS1411 / GPC1-gemcitabine-DNAh3.3-doxorubicin nanonucleic acid triaptamer + gemcitabine + doxorubicin integrated complex targeted chemotherapy agent has a unique mechanism of action and advantages. Below, we will examine in detail its mechanism of action, synergistic complex targeted delivery effect, doxorubicin active ingredient, gemcitabine slow-release effect, and comparative advantages.

[0155] 1. Mechanism of Action: This nanoparticle drug combines three aptamers (MUC1, AS1411, and GPC1), all of which target tumor-related biomarkers. MUC1 is a membrane-related glycoprotein overexpressed in many cancers, AS1411 is an oligonucleotide aptamer that binds to nucleolin receptors, and GPC1 is a cell surface glycoprotein associated with the growth and invasion of pancreatic cancer and other cancers. Through the synergy of the three aptamers, the drug accumulates in target cells, enhancing its therapeutic effect. 2. Synergistic multi-target delivery effect: Through the collaborative action of the three aptamers, the nanoparticles exhibit high specificity and selectivity, specifically binding and accumulating within the tumor microenvironment and tumor cells. This synergistic effect significantly increases the drug concentration in tumor tissue, enhancing therapeutic efficacy while reducing damage to normal tissue. 3. Slow-release effect of doxorubicin and gemcitabine: Doxorubicin is reductively dissociated and released under the low pH / high GSH environment within tumor cells, specifically binding to the "GC / CG" target of double-stranded DNA in the nucleus, inhibiting transcription and suppressing tumor cell division and proliferation. Gemcitabine dissociates by enzymatic cleavage and, as an alternative, inhibits DNA synthesis. These slow-release effects enable sustained release within tumor tissue, enhancing therapeutic efficacy and reducing side effects. 4,4) Comparative advantages: (1) High specificity: High specificity is achieved through the cooperation of the three aptamers, effectively reducing damage to normal tissue; (2) Enhanced therapeutic effect: Cooperative complex target delivery significantly increases the concentration in tumor tissue, enhancing the therapeutic effect. The combination of doxorubicin and gemcitabine suppresses the division and proliferation of tumor cells through the inhibition of DNA transcription and alternative inhibition of DNA synthesis, thereby enhancing the antitumor effect; (3) Reduced side effects: Due to the slow release of the doxorubicin drug and gemcitabine, they are released continuously in tumor tissue, reducing systemic toxicity and mitigating side effects; (4) Suppression of drug resistance: Since gemcitabine and doxorubicin have different mechanisms of action, the combination can suppress the development of drug resistance to monotherapy; (5) Broad applicability: Since MUC1, AS1411, and GPC1 aptamers all target biomarkers that are overexpressed in various cancers, this nanoparticle drug can be applied to many types of tumors.

[0156] Generally, the above-mentioned drug 21 has significant advantages and shows extremely high potential and application value in anti-tumor treatment due to its highly specific targeting, coordinated composite target delivery, slow release of doxorubicin raw drug and gemcitabine, and broad applicability.

[0157] The above various target chemical drug structures contribute to further improving the reliability of the target delivery of low-molecular chemical drugs and endowing them with better specificity. At the same time, by adopting the above target chemical drug structures, the loading stability and loading amount of low-molecular chemical drugs are further improved. By the above linking method, the drug size becomes more appropriate, and while it is taken into the cell membrane through cell surface receptor-mediated endocytosis, non-specific cell permeation can be avoided and filtration removal by the kidney can be avoided. Therefore, the advantageous particle size contributes to the improvement of pharmacokinetics, pharmacodynamics, biodistribution and toxicological distribution.

[0158] According to the second aspect of the present invention, a pharmaceutical composition comprising the target chemical drug and a pharmaceutically acceptable carrier and / or excipient is provided.

[0159] The above pharmaceutically selectable excipients may vary depending on the dosage form and / or administration mode and administration route of the pharmaceutical to be prepared, and can be reasonably selected from currently known pharmaceutical excipients. These include, but are not limited to, excipients such as pharmaceutically acceptable carriers, excipients or adjuvants. In addition, the target chemical drug can be formulated into different dosage forms to adapt to various administration routes, and the administration routes include oral administration, injection administration or transdermal administration, etc. Dosage forms of the pharmaceutical include capsules, tablets, oral liquids, injections or transdermal absorbents, etc.

[0160] Specifically, the target chemical agent of this application can be formulated into capsules, tablets, oral solutions, injections, or transdermal formulations according to methods known in the pharmaceutical industry. Preferably, the injection is an intravenous injection. When preparing capsules, tablets, or oral solutions suitable for oral administration, sucrose, lactose, galactose, corn starch, gelatin, microcrystalline cellulose, carboxymethylcellulose, etc., can be used as carriers or excipients. Furthermore, the pharmaceutical agent of this application can also be prepared into solutions and suspensions suitable for oral administration using methods and auxiliary components known in the pharmaceutical industry. When preparing solutions and suspensions suitable for extra-gastrointestinal administration (parenteral administration), distilled water, water for injection, isotonic sodium chloride solution, or glucose solution, or low-concentration (e.g., 1-100 mM) phosphate buffer (PBS) can be used as a carrier or diluent. These parenteral administration formulations may contain one or more other auxiliary components or additives, for example, ascorbic acid as an antioxidant or sodium benzoate as a preservative. These dosage forms may also contain other ingredients such as solvents, disintegrants, colorants, dispersants, or surfactants.

[0161] A third aspect of the present invention provides the use of the above-mentioned targeted chemical agent in the manufacture of a pharmacopoeia for tumor treatment. Furthermore, the tumor may be an androgenetic tumor, gynecological tumor, respiratory tumor, digestive tumor, hematological tumor, urological tumor, bone marrow tumor, neurological tumor, dermatological tumor, general surgical tumor, or endoscopic tumor; preferably, androgenetic tumors are prostate cancer, penile cancer, testicular tumor, or male urethral cancer; gynecological tumors are ovarian cancer, cervical cancer, endometrial cancer, uterine fibroid, vulvar cancer, or malignant hydatidiform mole; respiratory tumors are lung cancer, non-small cell lung cancer, small cell lung cancer, nasopharyngeal cancer, tracheal tumor, lung cancer metastasis, inflammatory pseudotumor, or radiation-induced lung cancer; digestive tumors are liver cancer, gastric cancer, colorectal cancer, gallbladder cancer, esophageal cancer, rectal cancer, pancreatic cancer, or colon cancer. Cancers and hematological tumors include leukemia, lymphoma, lymphosarcoma, or multiple myeloma; urological tumors include renal cancer, bladder cancer, or urinary tract cancer; bone marrow tumors include giant cell tumor, osteochondroma, or osteosarcoma; neurological tumors include brain tumors, meningiomas, cerebral tuberculoma, pituitary tumors, neuroblastoma, glioblastoma, or glioma; dermatological tumors include skin cancer or melanoma; general surgical tumors include breast cancer, lipoma, thyroid cancer, or thyroid tumor; and ossicular tumors include oral cancer, tongue cancer, laryngeal cancer, middle ear cancer, gingival cancer, or orbital tumors; preferably, the mode of administration during use is intratumor administration, intravenous administration, or intraperitoneal administration; preferably, the daily dose during use is 0.1 μg / kg to 100 mg / kg.

[0162] A fourth aspect of the present invention further provides a method for the prevention and / or treatment of cancer, the method comprising the steps of: providing the targeted chemical agent or pharmaceutical composition; and administering an effective amount of the targeted chemical agent or pharmaceutical composition to a tumor patient. Preferably, the tumor is an andrological tumor, gynecological tumor, respiratory tumor, digestive tumor, hematological tumor, urinary tract tumor, bone marrow tumor, neurological tumor, dermatological tumor, general surgical tumor, or endoscopic tumor; preferably, andrological tumors are prostate cancer, penile cancer, testicular tumor, or male urethral cancer; gynecological tumors are ovarian cancer, cervical cancer, endometrial cancer, uterine fibroid, vulvar cancer, or malignant hydatidiform mole; respiratory tumors are lung cancer, non-small cell lung cancer, small cell lung cancer, nasopharyngeal cancer, tracheal tumor, lung cancer metastasis, inflammatory pseudotumor, or radiation-induced lung cancer; digestive tumors are liver cancer, gastric cancer, colorectal cancer, gallbladder cancer, esophageal cancer, or rectal cancer. The tumors include pancreatic cancer or colon cancer, hematological tumors such as leukemia, lymphoma, lymphosarcoma, or multiple myeloma, urological tumors such as renal cancer, bladder cancer, or urinary tract cancer, bone marrow tumors such as giant cell tumor, osteochondroma, or osteosarcoma, neurological tumors such as brain tumor, meningioma, cerebral tuberculoma, pituitary tumor, neuroblastoma, glioblastoma, or glioma, dermatological tumors such as skin cancer or melanoma, general surgical tumors such as breast cancer, lipoma, thyroid cancer, or thyroid tumor, and ossicular tumors such as oral cancer, tongue cancer, laryngeal cancer, middle ear cancer, gingival cancer, or orbital tumor; preferably, the mode of administration during use is intratumorally, intravenously, or intraperitoneally.

[0163] The effective dose as used herein includes both a prophylactic effective dose and / or a therapeutic effective dose. A therapeutic effective dose is the amount that, at the required dose and duration, effectively achieves the desired therapeutic outcome (e.g., reduction in gastric or lung cancer). In specific embodiments, the dose may be adjusted to provide the optimal therapeutic response dose, and the therapeutic effective dose may vary depending on the following factors: the individual's disease state, age, sex, weight, and the ability of the formulation to elicit the desired response in the individual. The concept of a therapeutic effective dose also includes the meaning of an amount in which the therapeutic benefit outweighs its toxicity or adverse effects. A prophylactic effective dose is the amount that, at the required dose and duration, effectively achieves the desired preventive outcome (e.g., prevention or suppression of the development of acute leukemia, breast cancer, and / or ovarian cancer). The prophylactic effective dose can be determined based on the description of the therapeutic effective dose above. For any particular subject, the specific dose may be adjusted over time based on the individual's needs and the administerer's professional judgment.

[0164] A fifth aspect of the present invention also provides a method for producing a targeted chemical agent, characterized by comprising the following steps: forming a targeted nucleic acid carrier equipped with an optionally selected oligonucleotide effector and an optionally selected immunostimulant by sequence self-assembly; and linking a small molecule drug to the targeted nucleic acid carrier to obtain a targeted chemical agent. As described above, the targeted chemical agent comprises a targeted nucleic acid carrier and a small molecule drug supported on the targeted nucleic acid carrier, the targeted nucleic acid carrier comprising a nucleic acid carrier and a target molecule linked to the nucleic acid carrier; optionally further comprising an oligonucleotide effector and / or immunostimulant supported on the targeted nucleic acid carrier. During sequence self-assembly, the targeted nucleic acid carrier can be directly formed as a single chain during sequence design and synthesis, and then formed by self-assembly. It is also obvious to those skilled in the art that the small molecule drug can be linked to the targeted nucleic acid carrier during sequence design and synthesis, or by support onto the targeted nucleic acid carrier.

[0165] In the actual sequence self-assembly process, multiple single sequences can be set according to the base distribution of the target targeted nucleic acid carrier, and then self-assembly can be performed to obtain the targeted nucleic acid carrier. In a preferred embodiment, the step of forming a targeted nucleic acid carrier equipped with an optional oligonucleotide effector and an optional immunostimulant by the sequence self-assembly method includes dissolving at least three sequences comprising an optional oligonucleotide effector, an optional immunostimulant, and a target molecule in an assembly solution and performing a denaturation reaction to form a crude self-assembly product; and sequentially purifying, eluting, and evaporating the crude product to dryness to obtain the targeted nucleic acid carrier equipped with an optional oligonucleotide effector and an optional immunostimulant. By performing self-assembly with at least three sequences, the difficulty of assembly is reduced, and the length of the single sequences becomes more appropriate.

[0166] To make self-assembly more efficient, preferably, at least three sequences include at least the following three sequences: Sequence A: A sequence in which sequence A of an optional oligonucleotide effector, an optional immunostimulant, and an optional target molecule is directly linked to the end or linked via a base linkage bridge; Sequence B: A sequence in which sequence B of an optional oligonucleotide effector, an optional immunostimulant, and an optional target molecule is directly linked to the end or linked via a base linkage bridge; Sequence C: A sequence of an optional oligonucleotide effector Sequence C of a select oligonucleotide effector, an optional immunostimulant, and an optional target molecule is either directly linked or linked via a base linkage bridge; the molar ratio between sequences A, B, and C is 0.90-1.10:0.90-1.10:0.90-1.10, more preferably 1:1:1; if at least three of the sequences include sequences other than A, B, and C, the molar ratio between each sequence and sequence A is 0.90-1.10:0.90-1.10, more preferably 1:1. Controlling the molar ratio of the sequences within the above range improves assembly efficiency.

[0167] Preferably, if the at least three sequences include sequences other than sequences A, B, and C, these are considered complementary sequences, in which case a transition base segment is independently ligated to the 5′ or 3′ end of any of sequences A, B, and C (the transition base segment is ligated to at least one location on the 5′ or 3′ end of sequence A, the 5′ or 3′ end of sequence B, or the 5′ or 3′ end of sequence C, with one transition base segment corresponding to one complementary sequence), the complementary sequence exhibits a base-complementary structure with the transition base segment, and is ligated complementaryly during the self-assembly process; more preferably, the complementary sequence itself is the sense or antisense strand of an siRNA oligonucleotide effector and / or a miRNA oligonucleotide effector, and / or the complementary sequence is used to provide a support site for a small molecule target head, a target aptamer, or an immunostimulant. By using the complementary sequences described above, the length of the single sequence during the self-assembly process is shortened, making assembly more efficient and increasing the stability of the final drug.

[0168] More preferably, if the at least three sequences include complementary sequences, a complementary sequence carrying an optional low-molecular-weight target head, an optional target aptamer, or an optional immunostimulant is added to the assembly solution at the same time, and a denaturation reaction is carried out; in this case, the complementary sequence also functions as a carrier sequence for the low-molecular-weight target head, target aptamer, and immunostimulant, and can support them on the final target chemical during the self-assembly process.

[0169] In preferred embodiments, the assembly solution is an aqueous TMS solution, an aqueous sodium chloride solution, an aqueous magnesium chloride solution, or purified water; preferably, the temperature of the denaturation reaction is 80-99°C, more preferably 85-99°C, and even more preferably 90-99°C; preferably, after the completion of the denaturation reaction, the reaction system is cooled to a holding temperature for a holding reaction, and finally cooled to obtain the assembled product; here, the holding temperature is 70-50°C, more preferably 65-55°C, and even more preferably 63-57°C; the holding reaction time is 3-15 minutes, more preferably 3-10 minutes, and even more preferably 3-5 minutes; preferably, the cooling rate in the process of cooling the reaction system to a holding temperature is 2-10°C / min, more preferably 2-6°C / min, and even more preferably 2-3°C / min; preferably, the final cooling temperature is 0-25°C, more preferably 0-15°C, and even more preferably 0-4°C; preferably, the pH of the denaturation reaction is 5.4-8.8. Performing the denaturation reaction for the self-assembly of each sequence under the above processing conditions results in a higher yield and a more stable reaction.

[0170] Depending on the structural type of small molecule drugs, drug loading can be performed using the following methods: In the case of anthracycline or acridine chemicals, the step of loading the small molecule drug onto a targeted nucleic acid carrier includes: dissolving a targeted nucleic acid carrier comprising a small chemical molecule, a coupling agent, an optionally selected oligonucleotide effector, and an optionally selected immunostimulant in a first solvent, reacting under light-shielding conditions to specifically intercalate the small chemical molecule to the GC-adjacent base pairs of the targeted nucleic acid carrier to obtain a crude reaction product; sequentially crystallizing, washing, and evaporating the crude reaction product to obtain the targeted chemical; preferably, the coupling agent is formaldehyde and / or paraformaldehyde; preferably, the reaction temperature is 0 to 37°C, more preferably 4 to 20°C, and even more preferably 4 to 8°C; preferably, the reaction pH is 6 to 8; preferably, The reaction time is 24 to 96 hours, more preferably 48 to 72 hours, and even more preferably 72 hours; preferably, the first solvent is purified water and / or PBS buffer; preferably, the washing solvent used in the washing process is a halogenated alkane solvent, an ether solvent, or a benzene solvent, more preferably chloroform, dichloromethane, diethyl ether, toluene, or xylene; preferably, the solvent used in the crystallization process is a lower alcohol solvent, more preferably methanol, ethanol, or isopropanol; preferably, the average number of small molecule drugs loaded per targeted nucleic acid carrier equipped with an optional oligonucleotide effector and an optional immunostimulant is 1 to 50, more preferably 10 to 40, and even more preferably 20 to 35; In the case of pyrimidine-based chemicals, the small molecule target head, oligonucleotide effector, and immunostimulant are linked to the end bases of each sequence of the nucleic acid carrier via a base linkage bridge containing 1 to 10 bases, and the step of linking the small molecule drug to a targeted nucleic acid carrier equipped with an optional oligonucleotide effector and an optional immunostimulant includes the step of forming a targeted nucleic acid carrier equipped with an optional oligonucleotide effector and an optional immunostimulant by sequence self-assembly, linking the small molecule drug to at least one sequence of the nucleic acid carrier by substituting at least some of the bases in the base linkage bridge and / or in the form of extension bases and / or in the form of carrier backbone base substitution, and then completing the linkage of the small molecule drug by sequence self-assembly; preferably, the average number of small molecule drugs supported per targeted nucleic acid carrier molecule is 1 to 30, more preferably 10 to 20, and even more preferably 12 to 18; In the case of platinum-based chemicals, glutamate derivative chemicals, flavonoid chemicals, plant-derived drugs and their derivatives, folic acid-based chemicals, or salicylic acid-based chemicals, the step of linking a small molecule drug to a targeted nucleic acid carrier equipped with an optionally selected oligonucleotide effector and an optionally selected immunostimulant includes: a step of linking a linker to at least one sequence of the nucleic acid carrier equipped with an optionally selected oligonucleotide effector, an optionally selected immunostimulant, and a target molecule during the synthesis process of each sequence of the nucleic acid carrier to which the target molecule is linked, thereby obtaining a targeted nucleic acid carrier having a linker; and a step of reacting the targeted nucleic acid carrier having a linker with the small molecule drug in a second solvent, and then cooling, purifying, washing, and evaporating to dryness to obtain the target chemical; Preferably, if the small molecule drug itself has a succinimidyl group (NHS ester group), the small molecule drug is reacted with a targeted nucleic acid carrier having a linker in a second solvent; if the small molecule drug itself does not have a succinimidyl group, the method further includes the following before reacting in the second solvent: the small molecule drug is sequentially reacted with bis(2-hydroxyethyl) disulfide and N,N-diisopropylethylamine to introduce a succinimidyl group onto the small molecule drug. Preferably, the second solvent is purified water and / or PBS buffer.

[0171] To allow target aptamers to extend freely, maintain their three-dimensional structure, and preserve their specificity and affinity, fragment sequences containing 3 to 6 bases may be used as base-linking bridges, and these target aptamers may be linked to one end of a single-chain sequence. For example, an EGFR aptamer can be synthetically linked to the 5′ end of the b-chain using a base-linking bridge "TTTTT" (see the sequence structures of each drug described above for details). This method is obvious to those skilled in the art and will not be described in detail here. If the target aptamer is a different small molecule structure, it can also be directly synthetically linked to one end of a single chain by chemical modification, which is also a common practice in this art.

[0172] The present application will be described in more detail below based on specific embodiments, but these embodiments should not be construed as limiting the scope of the claims of this application.

[0173] Example 1 In this example, the targeted epirubicin drug Apt EGFR-DNA-epirubicin was synthesized. Step 1: Synthesis of the Apt EGFR-DNA target carrier, the single-stranded sequence of which is shown in Table 1 below:

[0174] [Table 1]

[0175] Note: The bolded portion indicates sequence A, sequence B, or sequence C in the nucleic acid carrier. The same applies below.

[0176] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) TMS buffer (Tris hydrochloride, sodium chloride, magnesium chloride); (5) TBM (Tris-borate-magnesium) buffer.

[0177] 2. Experimental method: Dissolve Array A, Array B, and Array C in TMS buffer at a molar ratio of 1:1:1, respectively. After mixing, adjust the pH of the reaction solution to 6, heat the mixture to 80 °C and hold for 5 minutes, then cool it to 70 °C at a rate of 10 °C / min and perform a heat preservation reaction for 15 minutes, and further cool it to 4 °C at a rate of 10 °C / min to obtain a crude product.

[0178] Apply the obtained product to an 8% (m / v) non-denaturing PAGE gel, perform electrophoresis at 4 °C and 100 V in TBM buffer to purify the carrier. Cut out the target band, elute it at 37 °C in DNA elution buffer, then perform ethanol precipitation overnight, and dry it under reduced pressure at low temperature to obtain a self-assembled product - EGFR-DNA target carrier.

[0179] Step 2: Loading of epirubicin 1. Experimental materials and reagents: (1) Apt EGFR-DNA target carrier prepared in Step 1; (2) DEPC-treated water; (3) PBS buffer (pH 7.4); (4) 4% formaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Absolute ethanol.

[0180] 2. Experimental method: (1) Dissolve epirubicin hydrochloride (0.79 mg, 1.354 μmol) in DEPC water (1.0 mL) and PBS buffer (1.25 mL), and add aqueous formaldehyde (0.25 mL) under ice water bath and mix. Mix the entire volume of this mixture with EGFR-DNA target carrier (1 mg) and react at 25 °C for 24 hours under light-shielding conditions, controlling the reaction pH to 6.

[0181] (2) Wash the reaction mixture with chloroform (10 mL x 3), add 25 mL of anhydrous ethanol to the aqueous phase and mix. Allow to stand in the dark at 4 °C for sufficient precipitation of the product (4 hours). Centrifuge (4000 rpm), transfer the supernatant, wash the solid product further with ethanol (50 mL), and remove the supernatant. Dry the residue under reduced pressure at low temperature to obtain a dark red solid product. This is the targeted epirubicin drug Apt EGFR-DNA-epirubicin.

[0182] (4) Calculation of load factor: 1. Prepare epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μM each, 100 μL each); 2. Dissolve Apt EGFR-DNA-epirubicin in 100 μL of PBS; 3. Dispense the standard solution and Apt EGFR-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature. 4. Using a microplate reader, measure the absorbance of epirubicin at 492 nm, create a calibration curve, and calculate the molar concentration of epirubicin in the loaded product; 5. Measure the absorbance of DNA at 260 nm using a spectrophotometer to obtain the mass concentration of the DNA carrier in each sample; 6. The loading rate is calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the targeted epirubicin drug Apt EGFR-DNA-epirubicin described above is approximately 23, and the measured actual average loading rate was 20.

[0183] Example 2 In this example, the targeted epirubicin drug Apt EGFR-4*Bio-DNA-epirubicin was synthesized.

[0184] Step 1: Synthesis of the Apt EGFR-4* Bio-DNA target carrier, the single-stranded sequence of which is shown in Table 2 below:

[0185] [Table 2]

[0186] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Sodium chloride solution; (5) TBM (Tris-borate-magnesium) buffer solution.

[0187] 2. Experimental method: Sequences A, B, and C are dissolved in aqueous sodium chloride in a molar ratio of 1.05:1:1.05. After mixing, the pH of the reaction solution is adjusted to 7. The mixture is heated to 85 °C and held for 5 minutes, then cooled to 50 °C at a rate of 4 °C / min and allowed to react for 10 minutes, and then cooled further to 10 °C at a rate of 4 °C / min to obtain the crude product.

[0188] The product is applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier is purified by electrophoresis in TBM buffer at 4 °C and 100 V. The target band is excised, eluted in DNA elution buffer at 37 °C, followed by ethanol precipitation overnight, and then dried under reduced pressure at low temperature to obtain the self-assembling product-Apt EGFR-4*Bio-DNA target carrier.

[0189] Step 2: Loading of epirubicin 1. Experimental materials and reagents: (1) Apt EGFR-4*Bio-DNA target carrier prepared in step 1; (2) DEPC treated water; (3) PBS buffer (5.8); (4) 4% paraformaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Anhydrous ethanol.

[0190] 2. Experimental method: (1) Dissolve epirubicin hydrochloride (0.79 mg, 1.354 μmol) in DEPC water (1.0 mL) and PBS buffer (1.25 mL), and add 4% paraformaldehyde aqueous solution (0.25 mL) and mix under an ice bath. Mix the entire volume of this mixture with EGFR-4 Bio-DNA target carrier (1 mg) and react at 20 °C for 48 hours under light-shielding conditions, controlling the reaction pH to 7.

[0191] (2) Wash the reaction mixture with chloroform (10 mL x 3), add 25 mL of anhydrous ethanol to the aqueous phase and mix, then stand in the dark at 4 °C to allow the product to precipitate completely (4 hours). Centrifuge (4000 rpm), transfer the supernatant, wash the solid product further with ethanol (50 mL), and remove the supernatant. Dry the residue under reduced pressure at low temperature to obtain a dark red solid product, namely EGFR-4Bio-DNA-epirubicin.

[0192] (4) Calculation of load factor: 1. Prepare epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μM each, 100 μL each); 2. Dissolve Apt EGFR-4*Bio-DNA-epirubicin in 100 μL of PBS; 3. Dispense the standard solution and Apt EGFR-4*Bio-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature. 4. Using a microplate reader, measure the absorbance of epirubicin at 492 nm, create a calibration curve, and calculate the molar concentration of epirubicin in the loaded product; 5. Measure the absorbance of DNA at 260 nm using a spectrophotometer to obtain the mass concentration of the DNA carrier in each sample; 6. The loading rate is calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the targeted epirubicin drug Apt EGFR-4*Bio-DNA-epirubicin described above is approximately 23, and the measured actual average loading rate was 20.

[0193] Example 3 In this example, the targeted epirubicin drug Apt AS1411-4*Bio-DNA-epirubicin was synthesized.

[0194] Step 1: Synthesis of the Apt AS1411-4*Bio-DNA target carrier. Its single-stranded sequence is shown in Table 3 below.

[0195] [Table 3]

[0196] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Aqueous solution of magnesium chloride; (5) TBM (Tris-borate-magnesium) buffer solution.

[0197] 2. Experimental method: Sequences A, B, and C are dissolved in aqueous magnesium chloride in a molar ratio of 1.05:1.05:1. After mixing, the pH of the reaction solution is adjusted to 8. The mixture is heated to 90 °C and held for 5 minutes, then cooled to 65 °C at a rate of 2 °C / min and allowed to hold for 5 minutes, and then cooled further to 5 °C at a rate of 2 °C / min to obtain the crude product.

[0198] The product is applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier is purified by electrophoresis in TBM buffer at 4 °C and 100 V. The target band is excised, eluted in DNA elution buffer at 37 °C, followed by ethanol precipitation overnight, and then dried under reduced pressure at low temperature to obtain the self-assembling product -Apt AS1411-4*Bio-DNA target carrier.

[0199] Step 2: Loading of epirubicin 1. Experimental materials and reagents: (1) Apt AS1411-4*Bio-DNA target carrier prepared in step 1; (2) DEPC treated water; (3) PBS buffer (6.5); (4) 4% paraformaldehyde; (5) Epirubicin; (6) Toluene; (7) Anhydrous ethanol.

[0200] 2. Experimental method: (1) Dissolve epirubicin hydrochloride (0.79 mg, 1.354 μmol) in DEPC water (1.0 mL) and PBS buffer (1.25 mL), and add 4% paraformaldehyde aqueous solution (0.25 mL) and mix under an ice bath. Mix the entire mixture with AS1411-4 Bio-DNA target carrier (1 mg) and react at 15 °C for 72 hours under light-shielding conditions, controlling the reaction pH to 8.

[0201] (2) Wash the reaction mixture with toluene (10 mL x 3), add 25 mL of anhydrous ethanol to the aqueous phase and mix, then stand in the dark at 4 °C to allow the product to precipitate completely (4 hours). Centrifuge (4000 rpm), transfer the supernatant, wash the solid product further with ethanol (50 mL), and remove the supernatant. Dry the residue under reduced pressure at low temperature to obtain a dark red solid product, namely AS1411-4Bio-DNA-epirubicin.

[0202] (4) Calculation of load factor: 1. Prepare epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μM each, 100 μL each); 2. Dissolve Apt AS1411-4*Bio-DNA-epirubicin in 100 μL of PBS; 3. Dispense the standard solution and Apt AS1411-4*Bio-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. Using a microplate reader, measure the absorbance of epirubicin at 492 nm, create a calibration curve, and calculate the molar concentration of epirubicin in the loaded product; 5. Measure the absorbance of DNA at 260 nm using a spectrophotometer to obtain the mass concentration of the DNA carrier in each sample; 6. The loading rate is calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the targeted epirubicin drug Apt AS1411-4*Bio-DNA-epirubicin is approximately 23, and the measured average loading rate was 20.

[0203] Example 4 In this example, the epirubicin-targeted drug Apt A1-Apt A15-DNA-epirubicin was synthesized.

[0204] Step 1: Synthesis of Apt A1-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 4 below:

[0205] [Table 4]

[0206] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-Borate-Mg) buffer.

[0207] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1.1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 5.4. The mixture was heated to 95°C and held for 5 minutes, then cooled to 55°C at a rate of 2°C / min, reacted at 55°C for 10 minutes, and then slowly cooled to 8°C at a rate of 2°C / min.

[0208] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the target carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised and eluted in DNA elution buffer at 37°C. Subsequently, ethanol precipitation was performed overnight, and the product was dried under low temperature and reduced pressure to obtain the self-assembling product Apt A1-Apt A15-DNA target carrier.

[0209] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt A1-Apt A15-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin; (6) ether; (7) Anhydrous ethanol.

[0210] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer (pH 6.6). 0.25 mL of 4% paraformaldehyde aqueous solution was added under ice water bath and mixed thoroughly. The entire resulting mixture was then mixed with Apt A1-Apt A15-DNA target carrier (1 mg). The reaction was carried out at 5°C for 96 hours under light-shielded conditions, during which the pH of the reaction solution was maintained between 6 and 8.

[0211] (2) The reaction mixture was washed with ether (10 mL x 3), then 25 mL of anhydrous ethanol was added to the aqueous layer and mixed. The mixture was left at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After that, the mixture was centrifuged (4000 rpm), the supernatant was removed, and the resulting solid product was further washed with 50 mL of ethanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid. This solid was designated as the epirubicin target drug Apt A1-Apt A15-DNA-epirubicin.

[0212] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt A1-Apt A15-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt A1-Apt A15-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical average loading rate of epirubicin for the epirubicin-targeted drug Apt A1-Apt A15-DNA-epirubicin was approximately 20, and the measured average loading rate was 18.

[0213] Example 5 In this example, the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin was synthesized.

[0214] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-2* Bio-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 5 below:

[0215] [Table 5]

[0216] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) TMS aqueous solution; (5) TBM (Tris-Borate-Mg) buffer.

[0217] 2. Experimental method: Sequences A, B, and C were each dissolved in TMS aqueous solution in a molar ratio of 1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 8.8. The mixture was heated to 99°C, held for 5 minutes, then cooled to 60°C at a rate of 2°C / min, reacted at 60°C for 5 minutes, and then further cooled to 2°C at a rate of 2°C / min to obtain the crude product.

[0218] The obtained product was applied to a series-connected SEC column, and the main fraction was separated. Desalting was performed on the column, and the mixture was dried under low temperature and reduced pressure to obtain the self-assembling product Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA target carrier.

[0219] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.8); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Dichloromethane; (7) Anhydrous ethanol.

[0220] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer (pH 6.8). 0.25 mL of 4% paraformaldehyde aqueous solution was added under ice water bath and mixed thoroughly. The entire resulting mixture was then mixed with Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0221] (2) The reaction mixture was washed with dichloromethane (10 mL x 3), then 25 mL of anhydrous ethanol was added to the aqueous layer and mixed. The mixture was left at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After that, the mixture was centrifuged (4000 rpm), the supernatant was removed, and the resulting solid product was further washed with 50 mL of ethanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid. This solid was designated as the epirubicin-targeting drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin.

[0222] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin was approximately 30, and the measured average loading rate was 25.

[0223] Example 6 In this example, the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin was synthesized.

[0224] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 6 below.

[0225] [Table 6]

[0226] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) TMS aqueous solution; (5) TBM (Tris-Borate-Mg) buffer.

[0227] 2. Experimental method: Sequences A, B, and C were each dissolved in TMS aqueous solution in a molar ratio of 1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 8.8. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 5°C / min, reacted at 65°C for 3 minutes, and then further cooled to 2°C at a rate of 5°C / min to obtain the crude product. Figure 1 shows the results of mass spectrometry performed on the crude targeted nucleic acid carrier after sequence assembly (measurement conditions are as follows: column: Biosep SEC-S2000, 5 μm, 7.8 × 300 mm, 2 columns connected in series; mobile phase: water (50 mM Tris-HCl, 100 mM sodium chloride, pH 8); detection wavelength: 260 nm; flow rate: 0.6 mL / min; column temperature: 35°C; sample preparation: dissolved in the fluid phase to a concentration of 2 mg / mL, gently shaken at 37°C for 30 minutes, then filtered through a 0.22 μm nylon filter; injection volume: 100 μL).

[0228] The obtained product was applied to SEC columns connected in series, the main fraction was separated, desalted by ultrafiltration, and dried under low temperature and reduced pressure. Figure 2 shows the HPLC chromatogram (SEC) results for the targeted nucleic acid carrier purified by HPLC.

[0229] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 7.0); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Anhydrous ethanol.

[0230] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer (pH 7.0). 0.25 mL of 4% paraformaldehyde aqueous solution was added under ice water bath and mixed thoroughly. The entire resulting mixture was then mixed with Apt AS1411-Apt EGFR-Apt A15-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0231] (2) The reaction mixture was washed with chloroform (10 mL x 3), then 25 mL of anhydrous ethanol was added to the aqueous layer and mixed. The mixture was left at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After that, the mixture was centrifuged (4000 rpm), the supernatant was removed, and the obtained solid product was further washed with 50 mL of ethanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid. This solid was designated as the epirubicin target drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin. Figure 3 shows the HPLC chromatogram (reverse phase) results of the target chemical drug crude after epirubicin loading. Furthermore, Figure 4 shows the measurement results of the HPLC chromatogram (reverse phase) of the target chemical drug crude purified by HPLC (reverse phase) column (test conditions are as follows: column: Aeris widepore XB-C18, 3.6 μm, 200 Å, 4.6 × 250 mm; mobile phase: A: water (1% HFIP, pH adjusted to 8 with TEA), B: acetonitrile; flow rate: 0.8 mL / min; detection wavelength: 260 nm; column temperature: 40°C): (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin was approximately 30, and the measured average loading rate was 25.

[0232] In this example, we further investigated how the final amount of Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin drug varied depending on the degree of excess epirubicin low molecular weight. Details are shown in Figures 5 to 7. Figure 5 is the HPLC reversed-phase chromatogram of the drug obtained when the molar ratio of epirubicin to carrier was 10:1, Figure 6 is the HPLC reversed-phase chromatogram of the drug obtained when the molar ratio of epirubicin to carrier was 20:1, and Figure 7 is the HPLC reversed-phase chromatogram of the drug obtained when the molar ratio of epirubicin to carrier was 30:1. From these figures, it was shown that (1) the distribution of epirubicin loading rate (DAR value) has a certain range, and (2) as the amount of epirubicin added increases, the characteristic peak area in the HPLC reversed-phase chromatogram clearly increases, and the epirubicin loading rate, i.e., the DAR value, approaches the theoretical loading rate.

[0233] Example 7 In this example, the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin was synthesized.

[0234] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 7 below.

[0235] [Table 7]

[0236] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0237] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 7. The mixture was heated to 85°C and held for 5 minutes, then cooled to 60°C at a rate of 2°C / min, reacted at 60°C for 5 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0238] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling product Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA target carrier.

[0239] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA target carrier prepared in Step 1; (2) DEPC water.

[0240] (3) PBS buffer (pH 6.8); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Methanol.

[0241] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and PBS buffer (1.25 mL). 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire resulting mixture was mixed with Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 6.

[0242] (2) The reaction mixture was washed with chloroform (10 mL x 3), then 25 mL of methanol was added to the aqueous phase and mixed. The mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After centrifugation (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of methanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin-targeting drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin.

[0243] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The average maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA-epirubicin was approximately 27, and the measured average loading rate was 23.

[0244] Example 8 In this example, the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin(1) was synthesized.

[0245] Step 1: Synthesis of Apt TfRA4-Apt AS1411-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 8 below:

[0246] [Table 8]

[0247] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0248] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, reacted at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0249] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling product Apt TfRA4-Apt AS1411-Apt A15-DNA target carrier (1).

[0250] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt TfRA4-Apt AS1411-Apt A15-DNA target carrier prepared in Step 1 (1); (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0251] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and PBS buffer (1.25 mL). 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire resulting mixture was mixed with AS1411-TfRA4-A15-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0252] (2) The reaction mixture was washed with chloroform (10 mL x 3), then 25 mL of isopropanol was added to the aqueous phase and mixed. The mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the resulting solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin-targeting drug AS1411-TfRA4-A15-DNA-epirubicin (1).

[0253] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin(1) was approximately 36, and the measured average loading rate was 30.

[0254] Example 9 In this example, the epirubicin-targeted drug Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin was synthesized.

[0255] Step 1: Synthesis of Apt TfRA3-Apt AS1411-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 9 below:

[0256] [Table 9]

[0257] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0258] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing the resulting solutions, the pH of the reaction mixture was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, reacted at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0259] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling product Apt TfRA3-Apt AS1411-Apt A15-DNA target carrier.

[0260] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt TfRA3-Apt AS1411-Apt A15-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0261] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and PBS buffer (1.25 mL). 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire resulting mixture was mixed with Apt AS1411-Apt TfRA3-Apt A15-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0262] (2) The reaction mixture was washed with chloroform (10 mL x 3), then 25 mL of isopropanol was added to the aqueous phase and mixed. The mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After centrifugation (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin-targeting drug Apt AS1411-Apt TfRA3-Apt A15-DNA-epirubicin.

[0263] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and the Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin solution into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drugs Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin was approximately 27, and the measured average loading rate was 25.

[0264] Example 10 In this example, the epirubicin-targeting drug CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA-epirubicin was synthesized.

[0265] Step 1: Synthesis of CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 10 below.

[0266] [Table 10] Note: All adjacent bases between the 1st to 24th bases at the 5' end of sequence A were modified with sulfur, and all adjacent bases between the 1st to 3rd bases at the 3' end were also modified with sulfur. All adjacent bases between the 1st to 3rd bases at the 5' and 3' ends of sequences B and C were modified with sulfur.

[0267] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0268] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0269] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA target carrier.

[0270] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) The CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6) (4) 4% paraformaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0271] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire mixture was mixed with CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0272] (2) The reaction mixture was washed with chloroform (10 mL x 3), and 25 mL of isopropanol was added to the aqueous phase and mixed. The mixture was then allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the resulting solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin-targeting drug CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA-epirubicin.

[0273] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug CPG2006(DNA)-Apt CD40-Apt PD-L1-DNA-epirubicin was approximately 25, and the measured average loading rate was 21.

[0274] Example 11 In this example, the epirubicin-targeting drug 2*CPG2006(DNA)-AptCD40-AptPD-L1-AptC12-DNA-epirubicin was synthesized.

[0275] Step 1: Synthesis of 2*CPG2006(DNA)-Apt CD40-Apt PD-L1-Apt C12-DNA target carriers The sequences of each single chain constituting the target carrier are shown in Table 11 below.

[0276] [Table 11]

[0277] Note: All adjacent bases between the 1st to 24th bases at the 5' end of sequences A and C were modified with sulfur. All adjacent bases between the 1st to 3rd bases at the 5' end of sequences B, D, and E were modified with sulfur.

[0278] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Array D; (5) Array E; (6) DEPC water; (7) TBM (Tris-magnesium borate) buffer solution.

[0279] 2. Experimental method: Sequences A, B, C, D, and E were each dissolved in DEPC water in a molar ratio of 1:1:1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0280] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and dried under low temperature and reduced pressure to obtain the self-assembling 2*CPG2006(DNA)-Apt CD40L-Apt PD-L1-Apt C12-DNA target carrier.

[0281] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) The 2*CPG2006(DNA)-Apt CD40-Apt PD-L1-Apt C12-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0282] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire mixture was mixed with 2*CPG2006(DNA)-Apt CD40-Apt PD-L1-Apt C12-DNA target carrier (1 mg). The reaction was allowed to proceed at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0283] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and then allowed to stand at 4°C under light-shielding conditions to precipitate the product sufficiently (4 hours). After centrifugation (4000 rpm) and transfer of the supernatant, the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin target drug 2*CPG2006(DNA)-Apt CD40-Apt PD-L1-Apt C12-DNA-epirubicin.

[0284] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. 2*CPG2006(DNA)-AptCD40-AptPD-L1-AptC12-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and 2*CPG2006(DNA)-AptCD40-AptPD-L1-AptC12-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug 2*CPG2006(DNA)-Apt CD40-Apt PD-L1-Apt C12-DNA-epirubicin was approximately 36, and the measured average loading rate was 30.

[0285] Example 12 In this example, the epirubicin-targeting drug CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA-epirubicin was synthesized.

[0286] Step 1: Synthesis of CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 12 below.

[0287] [Table 12] Note: All adjacent bases between the 1st to 24th bases at the 5' end of sequence A were modified with sulfide, and all adjacent bases between the 1st to 3rd bases at the 3' end were also modified with sulfide. All adjacent bases between the 1st to 3rd bases at both the 5' and 3' ends of sequence B were modified with sulfide, and all adjacent bases between the 19th to 21st bases at the 5' end of sequence B were also modified with sulfide. All adjacent bases between the 1st to 3rd bases at both the 5' and 3' ends of sequence C were modified with sulfide, and all adjacent bases between the 19th to 21st bases at the 5' end of sequence C were also modified with sulfide. All adjacent bases between the 1st to 3rd bases at both the 5' and 3' ends of sequence D were modified with sulfide, and all adjacent bases between the 19th to 21st bases at the 3' end of sequence D were also modified with sulfide. All adjacent bases between the 1st and 3rd bases at the 5' and 3' ends of sequence E were subjected to sulfide modification, and all adjacent bases between the 19th and 21st bases at the 3' end of sequence E were also subjected to sulfide modification. The C / U bases in the underlined RNA sequence are subjected to 2'F modification.

[0288] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Array D; (5) Array E; (6) DEPC water; (7) TBM (Tris-magnesium borate) buffer solution.

[0289] 2. Experimental method: Sequences A, B, C, D, and E are dissolved in DEPC water in a molar ratio of 1:1:1:1:1. After mixing, the pH of the reaction mixture is adjusted to 7. The mixture is heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, and the reaction is held for 15 minutes. After that, it is cooled to 4°C at a rate of 2°C / min.

[0290] The product is applied to an 8% (m / v) non-denaturing PAGE gel and purified by electrophoresis in TBM buffer at 4°C and 100V. The target band is excised, eluted in DNA elution buffer at 37°C, and then precipitated overnight in ethanol. The product is dried under low temperature and reduced pressure to obtain the self-assembled product-CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA target vector. Step 2: Epirubicin loading 1. Experimental materials and reagents: (1) The CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0291] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire mixture was mixed with CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0292] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After centrifugation (4000 rpm) and transfer of the supernatant, the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin-targeting drug CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA-epirubicin.

[0293] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug CPG2006(DNA)-Apt C12-CD47 siRNA-Apt PD-L1-PD-L1 siRNA-DNA-epirubicin was approximately 30, and the measured average loading rate was 26.

[0294] Example 13 In this example, the epirubicin-targeting drugs A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA-epirubicin were synthesized.

[0295] Step 1: Synthesis of A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA target carriers The sequences of each single chain constituting the target carrier are shown in Table 13 below.

[0296] [Table 13] Note: In sequence B, all adjacent pairs of bases between the 1st and 3rd bases at the 5' end were modified with sulfide. In sequence C, all adjacent pairs of bases between the 29th and 31st bases at the 5' end were modified with sulfide, and all adjacent pairs of bases between the 1st and 3rd bases at the 3' end were also modified with sulfide. In sequence D, all adjacent pairs of bases between the 1st and 3rd bases at the 3' end were modified with sulfide, and all adjacent pairs of bases between the 19th and 21st bases were also modified with sulfide. C / U bases in the underlined RNA sequences are modified with 2'F.

[0297] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Array D; (5) DEPC water; (6) TBM (Tris-magnesium borate) buffer.

[0298] 2. Experimental method: Sequences A, B, C, and D were each dissolved in DEPC water in a molar ratio of 1:1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0299] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA target carrier.

[0300] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0301] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire mixture was mixed with A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0302] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). After centrifugation (4000 rpm) and transfer of the supernatant, the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin target drug A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA-epirubicin.

[0303] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and A-miR-21-TGF-β1 siRNA-Apt TfRA4-Apt PD-L1-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using a microplate reader, and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drugs A-miR-21-TGF-β siRNA-Apt TfRA4-Apt PD-L1-DNA-epirubicin was approximately 33, and the measured average loading rate was 28.

[0304] Example 14 In this example, the epirubicin-targeting drugs Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA-epirubicin were synthesized.

[0305] Step 1: Synthesis of Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA target carriers The sequences of each single chain constituting the target carrier are shown in Table 14 below.

[0306] [Table 14] Note: All adjacent bases between the 1st to 3rd bases at the 3' end of sequence A were subjected to sulfide modification, and all adjacent bases between the 19th to 21st bases at the 3' end of sequence A were also subjected to sulfide modification. All adjacent bases between the 1st to 3rd bases at the 5' end of sequence B were subjected to sulfide modification, and all adjacent bases between the 19th to 21st bases at the 5' end of sequence B were also subjected to sulfide modification. All adjacent bases between the 1st to 3rd bases at the 3' end of sequence C were subjected to sulfide modification, and all adjacent bases between the 19th to 21st bases were also subjected to sulfide modification. All adjacent bases between the 1st to 4th bases at the 5' end of sequence D were subjected to sulfide modification, and all adjacent bases between the 1st to 3rd bases at the 3' end of sequence D were also subjected to sulfide modification, and all adjacent bases between the 19th to 21st bases at the 3' end of sequence D were also subjected to sulfide modification. In sequences A, B, and C, the C / U bases in the underlined RNA sequences are modified with 2'F. In sequence D, the first 44 bases are modified with 2'F on the C / U bases and 2'-OMe on the A / G bases, while the 45th to 64th bases are modified with 2'F on the C / U bases.

[0307] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) Array D; (5) DEPC water; (6) TBM (Tris-magnesium borate) buffer.

[0308] 2. Experimental method: Sequences A, B, C, and D were each dissolved in DEPC water in a molar ratio of 1:1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0309] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and dried under low temperature and reduced pressure to obtain the self-assembled Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA target carrier.

[0310] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0311] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice water, and the entire mixture was mixed with Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0312] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin target drug Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA-epirubicin.

[0313] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA-epirubicin was dissolved in 100 μL of PBS; 3. The standard solution and Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA-epirubicin were dispensed into PCR plates, heated at 85°C for 5 minutes, and then cooled to room temperature. 4. The absorbance of epirubicin at 492 nm was measured using an enzyme labeling analyzer (microplate reader), a standard curve was created, and the molar concentration of epirubicin in the onboard product was calculated. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drugs Apt IL-4Ra-CD47 siRNA-Apt TfRA4-PD-L1 siRNA-Apt GPC-1-DNA-epirubicin was approximately 32, and the measured average loading rate was 27.

[0314] Example 15 In this example, the gemcitabine-targeted drug Apt AS1411-Apt EGFR-Apt A15-DNA-gemcitabine was synthesized.

[0315] Apt AS1411-Apt EGFR-Apt A15-DNA-gemcitabine-targeted drug was synthesized, and its single-strand sequence is shown in Table 15 below:

[0316] [Table 15] Note: The above J represents gemcitabine, which is synthesized directly into the sequence during sequence synthesis in the form of base substitution and base extension, and corresponds to 36 gemcitabines being linked onto a single DNA target carrier.

[0317] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0318] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0319] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt AS1411-Apt EGFR-Apt A15-DNA-gemcitabine-targeted drug.

[0320] Example 16 In this example, epirubicin and the gemcitabine-targeted drug Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA epirubicin were synthesized.

[0321] Step 1: Synthesis of Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine-DNA target carrier Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine-DNA target carrier was synthesized, and its single-strand sequence is shown in Table 16 below.

[0322] [Table 16] Note: The above J represents gemcitabine, which is synthesized directly into the sequence during sequence synthesis in the form of base substitution and base extension. All adjacent bases of gemcitabine are sulfurized, which corresponds to 36 gemcitabines linked on a single DNA target carrier. All adjacent bases between the 1st and 2nd bases at the 5' end of sequences A, B, and C above have been sulfurized.

[0323] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0324] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0325] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA target carrier.

[0326] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0327] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice bath cooling, and the entire mixture was mixed with Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA target carrier (1 mg). The reaction was carried out at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0328] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was used as the Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA-epirubicin target drug.

[0329] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using an enzyme labeling analyzer (microplate reader), and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the above Apt MUC1-Apt AS1411-Apt GPC1-gemcitabine--DNA-epirubicin targeted drug is approximately 36, and the maximum theoretical loading rate of gemcitabine is 28. Based on the measurements, the average loading rate of epirubicin was 30, and the average loading rate of gemcitabine was 28.

[0330] Example 17 In this example, the 5-fluorouracil (5-FU) targeted drug Apt AS1411-Apt EGFR-Apt A15-RNA-5'fluorouracil was synthesized.

[0331] We synthesized the Apt AS1411-Apt EGFR-Apt A15-RNA-5' fluorouracil-targeted drug, and its single-chain sequence is shown in Table 17 below:

[0332] [Table 17] Note: The above N represents 5'-fluorouracil, which is directly synthesized into the sequence in the form of base substitution and base extension during sequence synthesis. All adjacent pairs of 5'-fluorouracil bases are sulfurized. This corresponds to 36 5'-fluorouracils being bound to a single DNA target carrier. For sequence A above, the first two bases at the 5' end are sulfurized; for sequence B, the first and second bases at the 5' and 3' ends are sulfurized; and for sequence C, the first and second bases at the 5' end are sulfurized. In RNA, C / U bases undergo 2'F modification, A / G bases undergo 2'-OMe modification, and adjacent bases between the first and second bases at the 5' and 3' ends undergo sulfurization. In the underlined RNA sequences, C / U bases undergo 2'F modification, and A / G bases undergo 2'-OMe modification.

[0333] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0334] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0335] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt AS1411-Apt EGFR-Apt A15-RNA-5' fluorouracil target drug.

[0336] Example 18 In this example, the paclitaxel-targeting drug Apt AS1411-Apt EGFR-Apt A15-DNA-paclitaxel was synthesized.

[0337] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 18 below.

[0338] [Table 18] Note: In the synthesis process of each sequence, when the Oligo synthesizer reaches a predetermined position (base-pair positions indicated by underlines in the table; there are 12 theoretical bond sites), a base phosphoramidite monomer having a long-chain primary amine linker (N4-benzoyl group-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine nucleoside-3'-(6-aminohexyl-2-hydroxymethyl)-N,N'-diisopropylphosphoramidite) is introduced via a programmed Oligo synthesizer, and solid-phase synthesis is performed. After the reaction is complete, the subsequent program is executed to complete the synthesis and modification of the entire single-chain. All adjacent bases between the 1st and 2nd bases at the 5' and 3' ends of sequences A, B, and C were subjected to sulfurization modification. For sequence A, amino group (-NH2) modification was applied to the 35th, 39th, 54th and 59th bases counting from the 5' end; for sequence B, amino group (-NH2) modification was applied to the 50th, 55th, 66th and 70th bases counting from the 5' end; and for sequence C, amino group (-NH2) modification was applied to the 24th and 28th bases counting from the 5' end.

[0339] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0340] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0341] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt AS1411-Apt EGFR-Apt A15-DNA target carrier.

[0342] Step 2: Paclitaxel modification 1.2 g (0.004 mol) of triphosphorus was dissolved in 25 mL of dichloromethane, and under stirring at room temperature, a solution of 8.5 g (0.01 mol) of paclitaxel and 2.45 g (0.02 mol) of 4-dimethylaminopyridine dissolved in 100 mL of dichloromethane was gradually added dropwise over approximately 30 minutes. After the addition was complete, the reaction was continued for another 10 minutes, and then a solution of 1.53 g (0.01 mol) of bis(2-hydroxyethyl) disulfide dissolved in 10 mL of dichloromethane was added, and the reaction was continued with stirring at room temperature for another 30 minutes. The solvent was evaporated to dryness, and 40 mL of ethyl acetate was added to the resulting residue, which was then washed sequentially with 10% KHSO4 aqueous solution, 5% NaHCO3 aqueous solution, and saturated sodium chloride aqueous solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated to dryness to obtain 9.1 g of paclitaxel-hydroxyethyl disulfide (yield 88%). Recrystallization with methanol yielded 8.2 g of a white crystalline solid.

[0343] 8.2 g (0.008 mol) of paclitaxel-hydroxyethyl disulfide was dissolved in 50 mL of dichloroethane, and a solution of 1.5 g (0.012 mol) of N,N-diisopropylethylamine dissolved in 10 mL of dichloroethane was added dropwise under stirring at room temperature. After the addition was complete, the mixture was reacted under reflux for 3 hours. After the reaction was complete, the mixture was cooled to room temperature, 40 mL of water was added and stirred for 5 minutes, then transferred to a separatory funnel and allowed to stand to separate the layers. The lower organic phase was separated, and the solvent was evaporated to dryness to obtain 7.7 g of paclitaxel disulfide succinimide ester (yield 81%).

[0344] Step 3: Paclitaxel installation 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Paclitaxel disulfide succinimide ester prepared in step 2; (6) Chloroform; (7) Isopropanol.

[0345] 2. Experimental method: The Apt AS1411-Apt EGFR-Apt A15-DNA target carrier and paclitaxel disulfide succinimide ester were added to 30 times the volume of paclitaxel in water in a molar ratio of 1:12, and the mixture was reacted at room temperature for 48 hours. After the reaction was complete, the mixture was cooled, and the solvent was removed by vacuum distillation until it was almost dry. Ethanol was added to the resulting residue, and the mixture was stirred and dispersed. The residue was filtered and washed with ethanol. The filtered cake was dried under vacuum to obtain Apt AS1411-Apt EGFR-Apt A15-DNA-paclitaxel.

[0346] Example 19 In this example, the meitansinoid-targeting drug Apt AS1411-Apt EGFR-Apt A15-DNA-meitansinoid was synthesized.

[0347] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 19 below.

[0348] [Table 19] Note: In the synthesis process of each sequence, when the Oligo synthesizer reaches a predetermined position (base pair position shown underlined in the table; there are 6 theoretical bond sites), a base phosphoramidite monomer having a long-chain primary amine linker (N4-benzoyl group-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine nucleoside-3'-(6-aminohexyl-2-hydroxymethyl)-N,N'-diisopropylphosphoramidite) is introduced via a program to perform solid-phase synthesis. After the reaction is complete, the subsequent program is executed to complete the synthesis and modification of the entire single chain. All adjacent bases between the 1st and 2nd bases at the 5' and 3' ends of sequences A, B, and C were modified with sulfur. For sequence A, amino group (-NH2) modification was applied to the 43rd and 61st bases counting from the 5' end; for sequence B, amino group (-NH2) modification was applied to the 48th and 63rd bases counting from the 5' end; and for sequence C, amino group (-NH2) modification was applied to the 24th and 43rd bases counting from the 5' end.

[0349] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0350] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0351] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt AS1411-Apt EGFR-Apt A15-DNA target carrier.

[0352] Step 2: Installation of Maytansinoid 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Maytansinoid SMCC-DM1: C51H66ClN5O16S, Molecular weight 1072.61, CAS No.: 1228105-51-8; [ka] (6) Chloroform; (7) Isopropanol.

[0353] 2. Experimental method: The Apt AS1411-Apt EGFR-Apt A15-DNA target carrier and the meitansinoid SMCC-DM1 were added to 30 times the volume of water in a molar ratio of 1:6, and the mixture was reacted at room temperature for 48 hours. After the reaction was complete, the mixture was cooled, and the solvent was removed by vacuum distillation until it was almost dry. Ethanol was added to the resulting residue, which was stirred and dispersed, filtered, and washed with ethanol. The filtered cake was dried under vacuum to obtain Apt AS1411-Apt EGFR-Apt A15-DNA-meitansinoid.

[0354] Example 20 In this example, the epirubicin-targeting drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin was synthesized.

[0355] Step 1: Synthesis of Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA target carrier The sequences of each single chain constituting the target carrier are shown in Table 20 below.

[0356] [Table 20]

[0357] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0358] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 85°C and held for 5 minutes, then cooled to 60°C at a rate of 2°C / min, the reaction was held at 60°C for 5 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0359] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and then dried under low temperature and reduced pressure to obtain the self-assembling Apt AS1411-Apt EGFR-Apt A15-2*Bio-AmiR-21-DNA target carrier.

[0360] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA target carrier prepared in Step 1; (2) DEPC water; (3) PBS buffer (pH 6.8); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Methanol.

[0361] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice bath cooling, and the entire mixture was mixed with Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA target carrier (1 mg). The reaction was allowed to proceed at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 6.

[0362] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of methanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of methanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin target drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin.

[0363] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using an enzyme labeling analyzer (microplate reader), and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The average maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-epirubicin was approximately 27, and the measured average loading rate was 23.

[0364] Example 21 In this example, the epirubicin-targeted drug Apt TfRA4-AptAS1411-Apt A15-DNA-epirubicin(2) was synthesized.

[0365] Step 1: Synthesis of Apt TfRA4-AptAS1411-Apt A15-DNA target carrier (2) The sequences of each single chain constituting the target carrier are shown in Table 21 below.

[0366] [Table 21]

[0367] 1. Experimental materials and reagents: (1) Array A; (2) Array B; (3) Array C; (4) DEPC water; (5) TBM (Tris-magnesium borate) buffer solution.

[0368] 2. Experimental method: Sequences A, B, and C were each dissolved in DEPC water in a molar ratio of 1:1:1. After mixing, the pH of the reaction solution was adjusted to 7. The mixture was heated to 99°C and held for 5 minutes, then cooled to 65°C at a rate of 2°C / min, the reaction was held at 65°C for 15 minutes, and then further cooled to 4°C at a rate of 2°C / min.

[0369] The obtained product was applied to an 8% (m / v) non-denaturing PAGE gel, and the carrier was purified by electrophoresis in TBM buffer at 4°C and 100 V. The target band was excised, washed in DNA washing buffer at 37°C, followed by ethanol precipitation overnight, and dried under low temperature and reduced pressure to obtain the self-assembled Apt TfRA4-AptAS1411-Apt A15-DNA target carrier (2).

[0370] Step 2: Epirubicin installation 1. Experimental materials and reagents: (1) Apt TfRA4-AptAS1411-Apt A15-DNA target carrier prepared in Step 1 (2); (2) DEPC water; (3) PBS buffer (pH 6.6); (4) 4% paraformaldehyde aqueous solution; (5) Epirubicin hydrochloride; (6) Chloroform; (7) Isopropanol.

[0371] 2. Experimental method: (1) 0.79 mg (1.354 μmol) of epirubicin hydrochloride was precisely weighed and dissolved in 1.0 mL of DEPC water and 1.25 mL of PBS buffer. 0.25 mL of 4% paraformaldehyde aqueous solution was added and mixed under ice bath cooling, and the entire mixture was mixed with AS1411 TfRA4 A15 DNA target carrier (1 mg). The reaction was allowed to proceed at 4°C for 72 hours under light-shielding conditions, maintaining the pH of the reaction solution at 7.

[0372] (2) The reaction mixture was washed with chloroform (10 mL x 3), 25 mL of isopropanol was added to the aqueous phase and mixed, and the mixture was allowed to stand at 4°C under light-shielding conditions to allow the product to precipitate sufficiently (4 hours). The mixture was centrifuged (4000 rpm), the supernatant was transferred, and the obtained solid product was further washed with 50 mL of isopropanol. The supernatant was removed, and the residue was dried under reduced pressure at low temperature to obtain a dark red solid product. This solid was designated as the epirubicin target drug Apt TfRA4-AptAS1411-Apt A15-DNA-epirubicin(2).

[0373] (4) Calculation of installation rate: 1. Epirubicin-PBS standard solutions of known concentrations (2 μM, 4 μM, 6 μM, 8 μM, 10 μM, 100 μL each) were prepared; 2. Apt TfRA4-AptAS1411-Apt A15-DNA-epirubicin was dissolved in 100 μL of PBS; 3. Dispense the standard solution and Apt TfRA4-AptAS1411-Apt A15-DNA-epirubicin into PCR plates, heat at 85°C for 5 minutes, and then cool to room temperature; 4. The absorbance of epirubicin at 492 nm was measured using an enzyme labeling analyzer (microplate reader), and a standard curve was created to calculate the molar concentration of epirubicin in the onboard product. 5. The absorbance of DNA at 260 nm was measured using a spectrophotometer, and the mass concentration of DNA carriers in each sample was determined. 6. The loading rate was calculated based on the measured molar concentration of epirubicin and the mass concentration of the DNA carrier. The maximum theoretical loading rate of epirubicin for the epirubicin-targeted drug Apt TfRA4-AptAS1411-Apt A15-DNA-epirubicin was approximately 36, and the measured average loading rate was 31.

[0374] Performance characteristics: 1. In vivo efficacy study of the epirubicin-targeted drug Apt EGFR-DNA-epirubicin prepared in Example 1 in an N87 mouse gastric cancer model: Laboratory animals Species and strain: Balb / c nude Age: 6-8 weeks Gender: male or female Weight: 18~22g Experimental method 1. Cell Culture: Tumor cells were cultured in RPMI-1640 medium containing 10% inactivated fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin, and 2 mM glutamine in an incubator at 37°C and 5% CO2. The cells were passed every 3-4 days once they nearly saturated the bottom of the flask, and tumor cells in the logarithmic growth phase were used for inoculation into the tumor cell culture.

[0375] 2. Inoculation and grouping of tumor cells: Tumor cells, 1 × 10⁷, resuspended in PBS:Matrigel, were inoculated subcutaneously into the right flank of test animals. The tumor was approximately 80–100 mm in size. 3 When the levels increased to a certain point, the patients were divided into groups and administration was initiated. The specific administration scheme is shown in the table below (PBS and epirubicin active pharmaceutical ingredient were set as controls).

[0376] 3. Administration scheme: [Table 22]

[0377] Note: If the animal's body weight decreases by 20% during the study, discontinue administration. Restart administration after the body weight has recovered.

[0378] The dosage was 200 μL / 20 g mouse, and the specific dosage was calculated based on the mouse's body weight. The observation period was 60 days (adjusted depending on the condition of the mouse and the tumor).

[0379] Animal Husbandry and Management Animal Care: Laboratory animals were kept in a temperature- and humidity-controlled, air-conditioned chamber of SPF grade, using independent ventilation cages (IVCs), with 3 animals per cage.

[0380] Temperature and Humidity: Room temperature was maintained at (23±3)°C, and humidity was kept within the range of 40-70%.

[0381] Cages: The cages were made of polycarbonate and measured 370 mm × 155 mm × 135 mm. Clean bedding made from autoclaved soft corn cobs was used and replaced twice a week. Each cage was labeled with a cage label indicating the number of animals, sex, strain, date of arrival, group, and start date of the experiment.

[0382] Feed and drinking water: SPF-grade mouse solid feed was used as feed and sterilized by Co-60 irradiation. Drinking water was purified by ultrafiltration and subjected to autoclaving. The animals were given free access to the autoclaved sterile feed and sterile drinking water.

[0383] Animal numbering: Individual identification was performed using the ear-punch method.

[0384] Test indicators 1. Test indicators a. Tumor volume: The longest and shortest diameters of the tumor were measured twice a week using calipers. The formula for calculating tumor volume was as follows: Volume = 0.5 × Longest diameter × Shortest diameter 2 .

[0385] b. Response after animal administration: Mouse body weight was weighed simultaneously with tumor volume measurement. The relationship between changes in mouse body weight and the timing of administration was recorded, and the survival status and health status of the mice, specifically their activity level and feeding status during the administration period, were observed.

[0386] c. Tumor weight measurement and imaging: After administration, mice were euthanized, images were taken, and tissue samples were collected. These samples were then fixed or cryopreserved as necessary.

[0387] a) Each animal was photographed after euthanasia, including the tumor; b) For each animal, the subcutaneous tumor was removed and its weight was measured; c) All tumors were coded according to the grouping table, arranged on a whiteboard, and a group photograph was taken. After longitudinal sectioning all tumors, they were again coded according to the grouping table, arranged on a whiteboard, and a group photograph was taken; d) The tumor was cryopreserved or formalin-fixed as instructed; d. Organ collection, photography, measurement, and imaging by autopsy: a) For each animal, the heart, liver, spleen, lungs, and kidneys were collected simultaneously, their weight was measured and recorded, and the organ / body weight ratio was calculated. Metastatic lesions visible to the naked eye were marked and photographed. The organs were fixed or cryopreserved, and their condition was recorded.

[0388] b) One mouse was selected from each test group, and its heart, liver, spleen, lungs, kidneys, and tumors were collected and imaged using the fluorescence wavelength of epirubicin to detect drug residues.

[0389] 2. Drug evaluation indicators: a. Tumor growth inhibition rate (%) Tumor growth inhibition rate (%) = (1- T / C) × 100% Here, the relative tumor growth rate (T / C) is shown as T / C = RTV of the treatment group / RTV of the control group, where RTC is the relative tumor volume.

[0390] Effectiveness was determined when the tumor growth suppression rate was 60% or higher and statistically p < 0.05.

[0391] b. Tumor volume ratio (T / C) of treatment group / control group (%) The ratio of tumor volume in the treatment group to the control group (T / C) is calculated as T / C (%) = RTV of the treatment group / RTV of the control group × 100%.

[0392] Efficacy evaluation criteria: If T / C(%) > 40%, it was considered ineffective, and if T / C(%) ≤ 40% and p < 0.05, it was judged as effective.

[0393] c. Aneurysm weight suppression rate IRTW The calculation formula was as follows: Tumor weight reduction rate IRTW (%) = (W solvent control group - W treatment group) / W solvent control group × 100, where W represents tumor weight. An IRTW% exceeding 60% was considered effective.

[0394] The test results are shown in the table below: [Table 23] II. In vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2*Bio-DNA-epirubicin prepared in Example 5 in an N87 mouse gastric cancer model: Laboratory animals Species and strain: Balb / c nude Age in weeks: 5 weeks Gender: Male Weight: Approximately 16 g Experimental method 1. Cell culture and preparation: After thawing the cells, they were seeded and cultured in the corresponding medium [1640 basal medium + 10% FBS + 1% P / S (penicillin / streptomycin)].

[0395] Subculturing method: a) Discard the culture medium, lightly wash the cells with PBS, and wash twice.

[0396] b) An appropriate amount of 0.25% trypsin was added, and the cells were digested in a 37°C incubator for approximately 1-2 minutes. After the cells had clearly contracted, complete medium (1640 basal medium + 10% FBS + 1% P / S) was added to neutralize the trypsin. The cells were then gently pipetteed to detach them, and the cells were subcultured at the specified ratio.

[0397] c) Subculturing ratio: 1:2 to 1:4; in this laboratory, the subculturing ratio was 1:3.

[0398] d) Passaging frequency: Passaging was performed every 2-3 days, depending on the cell seeding density and growth status.

[0399] Cell retrieval: a) Cells in the logarithmic growth phase (confluence of approximately 80-90% is desirable) were collected.

[0400] b) After digesting the cells with trypsin, complete medium was added to stop the digestion, and the cells were collected in a centrifuge tube and centrifuged at 1000 rpm for 5 minutes.

[0401] c) Washed twice with pre-cooled PBS and centrifuged.

[0402] d) The cell precipitate was suspended in PBS or serum-free medium and adjusted to an appropriate concentration of 2 × 10⁷ cells / mL.

[0403] 2, Inoculation: Inoculation site: subcutaneous on the back of the right axilla Inoculation dose: 3 × 10⁶ cells / mice, administered to 30 mice.

[0404] Inoculation volume: 0.1 mL Inoculation method: cell inoculation N87 cells were thawed and cultured to a predetermined number, then the cell concentration was adjusted to 3 × 10⁷ cells / mL using sterile PBS and kept on ice. 0.1 mL of the cell suspension was aspirated using a sterile disposable 1 mL syringe and inoculated subcutaneously into the right axilla of mice. The day of inoculation was designated as D0.

[0405] Vaccination procedure: a) Before inoculation, the tumor suspension was thoroughly pipetted using a micropipette to prevent cell aggregation and a decrease in viability.

[0406] b) The mouse's neck skin was pinched between the thumb and index finger of the left hand, and its tail was held in place with the little finger.

[0407] c) An appropriate amount of tumor suspension was collected using a disposable 1 mL syringe and injected subcutaneously into mice. During administration, the needle tip was inserted slightly deeper (approximately 1 cm) subcutaneously to prevent leakage of the cell suspension from the injection site after injection. 3. Administration scheme: [Table 24] Test indicators 1. General clinical observation: During the study period, the clinical condition of the mice was observed daily. Observations included the ulceration and rupture status of tumor nodules, the animals' mental state, coat condition, behavior, feeding, and excretion.

[0408] 2. Weight: Mouse body weight was measured three times a week, and the data was recorded and analyzed for changes. The relative weight gain curves for each group are shown in Figure 8.

[0409] 3. Tumor volume (V): Tumor volume was measured three times a week using calipers, and the longest and shortest diameters of the tumor were recorded. Tumor volume was calculated using the following formula: Volume (V) = 0.5 × Longest diameter (a) × Shortest diameter (b) 2 .

[0410] 4. Relative tumor volume (RTV) The RTV was calculated using the following formula: RTV = Vt / V0 Here, V0 represents the tumor volume measured at the time of group administration (i.e., D0), and Vt represents the tumor volume at each measurement time point.

[0411] 5. Tumor growth suppression rate: The tumor growth inhibition rate (%) was calculated using the following formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100%. T / C = Treatment group RTV / Control group RTV If the tumor growth inhibition rate was 60% or higher, and statistical testing showed that the tumor volume in the treatment group was significantly lower than that in the solvent control group (P<0.05), the treatment was judged to be effective and to have a significant inhibitory effect on tumor growth.

[0412] 6. Aneurysm weight suppression rate IRTW IRTW (%) = (W solvent control group - W treatment group) / W solvent control group × 100 Here, W represents tumor weight. An IRTW% exceeding 60% was considered effective.

[0413] The test results are shown in Table 25, and the tumor growth curves for each group are shown in Figure 9.

[0414] [Table 25]

[0415] The results for tumor weight and tumor weight reduction rate (IRTW%) are shown in Table 26, and the changes in tumor volume for each group are shown in Figure 10.

[0416] [Table 26]

[0417] 3. Comparison of in vivo efficacy studies in the N87 mouse gastric cancer model of the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2Bio-DNA-epirubicin prepared in Example 5 and the epirubicin-targeted drug Apt AS1411-Apt EGFR-Apt A15-2Bio-AmiR-21-DNA-epirubicin prepared in Example 7: Laboratory animals Species and strain: Balb / c nude Age: 6-8 weeks Gender: Male Weight: 17~19 g Animal rearing location: The animals were reared in isolators in an SPF-grade barrier facility environment at the Institute of Pharmaceutical Sciences, Chinese Academy of Medical Sciences, in accordance with the facility's standards. Six animals were kept per cage, and the animal room had good lighting, good ventilation and air conditioning, and was maintained at a room temperature of 18-25°C and a relative humidity of 50-70%.

[0418] Experimental method 1. Inoculation and Grouping: BALB / c-nu mice (male, body weight 17.0-19.0 g) were used as the mouse gastric cancer N87 model. N87 tumor tissue was excised under sterile conditions, cut into tumor fragments measuring 2.0 mm × 2.0 mm × 2.0 mm, and uniformly inoculated subcutaneously into the left axillary dorsal region of naked mice. Day of inoculation was designated as D0. The tumor volume was approximately 250.0 mm³. 3 At the point when the tumor growth rate reached (D6), animals with poor tumor growth were excluded, and the animals were randomly divided into groups based on tumor volume. Treatment was started on the day of group division.

[0419] 2. Administration scheme: [Table 27]

[0420] Test indicators 1. General clinical observation: During the study period, the clinical condition of the mice was observed daily. Observations included the rupture and ulceration status of tumor nodules, the animals' mental state, coat condition, behavior, and feeding and excretion status.

[0421] 2. Weight: The weight of the mice was weighed three times a week, and the data was recorded and the changes were analyzed. The mouse weight change curve is shown in Figure 11.

[0422] 3. Tumor volume (V): The tumor volume was measured three times a week using calipers, and the longest and shortest diameters of the tumor were recorded. The volume was calculated using the following formula:

[0423] Volume (V) = 0.5 × Major axis (a) × Minor axis (b) 2 .

[0424] 4. Relative tumor volume (RTV): The RTV was calculated using the following formula: RTV = Vt / V0 Here, V0 represents the tumor volume measured at the time of group administration (i.e., D0), and Vt represents the tumor volume at each measurement time point.

[0425] 5. Tumor growth suppression rate: The tumor growth inhibition rate (%) was calculated using the following formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100% T / C = Treatment group RTV / Control group RTV Effectiveness was determined when the tumor growth inhibition rate was 60% or higher, and statistical testing showed that the tumor volume in the treatment group was significantly lower than that in the solvent control group (P<0.05), meaning that the treatment had a significant inhibitory effect on tumor growth.

[0426] 6. Aneurysm weight suppression rate IRTW The tumor weight reduction rate (IRTW) was calculated using the following formula: Tumor weight reduction rate (IRTW) = (W solvent control group - W treatment group) / W solvent control group × 100. Here, W represents tumor weight. An IRTW% of over 60% was considered effective.

[0427] During the study period, tumor growth was strictly monitored for all animals, and tumor measurements were taken twice a week. The results were recorded, and the tumor growth suppression rate was calculated. The results of the tumor growth suppression rate study are shown in Table 28, the tumor volume change curves are shown in Figure 12, and the relative tumor volume change curves are shown in Figure 13.

[0428] [Table 28]

[0429] The results of the trial for the tumor volume ratio (T / C) between the treatment group and the control group are shown in the table below: [Table 29]

[0430] Forty-one days after N87 vaccination, all animals were euthanized by cervical dislocation, and the tumors were dissected and measured. The tumor weight reduction rate (IRTW%) is shown below: [Table 30]

[0431] Based on the above data, the tumor growth inhibition rate (based on tumor weight and tumor volume) at the end of the study (D41) was less than 60% for epirubicin administered via tail vein, and the T / C ratio was greater than 40%. On the other hand, for the targeted therapies Apt AS1411-Apt EGFR-Apt A15-2Bio-DNA-epirubicin and Apt AS1411-Apt EGFR-Apt A15-2Bio-AmiR-21-DNA-epirubicin administered via tail vein, the tumor growth inhibition rate (based on tumor weight and tumor volume) at the end of the study (D41) was greater than 60% for both, and the T / C ratio was less than 40% for both. This suggests that the targeted drugs Apt AS1411-Apt EGFR-Apt A15-2Bio-DNA-epirubicin and Apt AS1411-Apt EGFR-Apt A15-2Bio-AmiR-21-DNA-epirubicin, administered intravenously, have a significant growth-inhibiting effect on N87.

[0432] IV. In vivo efficacy study of the epirubicin-targeted drug Apt AS1411-4*Bio-DNA-epirubicin prepared in Example 3 in an A549 mouse lung cancer model: Laboratory animals Species and strain: Balb / c nude Age: 6-7 weeks Gender: Male Weight: 16~18 g Experimental method 1. Inoculation and grouping: Balb / c-nu mice (male, body weight 16.0-18.0 g) were used as the mouse lung cancer A549 model. A549 tumor cells were collected under sterile conditions, the cell concentration was adjusted to 1 × 10⁷ cells / mL with sterile saline, and 0.2 mL of this solution was inoculated subcutaneously into the axillary dorsal region of naked mice. Tumors with a diameter of 1000-1500 mm 3 After growing to a certain extent, the tumor was excised under sterile conditions, cut into 2.0 mm × 2.0 mm × 2.0 mm tumor fragments, and uniformly inoculated subcutaneously into the axillary dorsum of naked mice. The day of inoculation was designated as D0. The tumor volume was approximately 100.0 mm³. 3At the point when the tumor growth rate reached (D5), animals with poor tumor growth were excluded, and the animals were randomly divided into groups based on tumor volume. Treatment was started on the day of group division.

[0433] 2. Administration scheme: [Table 31]

[0434] Test indicators 1. General clinical observation: During the study period, the clinical condition of the mice was observed daily. Observations included the rupture and ulceration status of tumor nodules, the animals' mental state, coat condition, behavior, feeding, and excretion.

[0435] 2. Weight: We weighed the mice twice a week, recorded the data, and analyzed the changes.

[0436] 3. Tumor volume (V): Tumor volume was measured twice a week using calipers, and the longest and shortest diameters of the tumor were recorded. The formula for calculating volume was as follows: Volume (V) = 0.5 × Longest diameter (a) × Shortest diameter (b) 2 .

[0437] 4. Relative tumor volume (RTV) The RTV was calculated using the following formula: RTV = Vt / V0 Here, V0 represents the tumor volume measured at the time of group administration (i.e., D0), and Vt represents the tumor volume at each measurement time point.

[0438] 5. Tumor growth suppression rate: The tumor growth inhibition rate (%) was calculated using the following formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100% T / C = Treatment group RTV / Control group RTV Effectiveness was determined when the tumor growth inhibition rate was 60% or higher, and statistical testing showed that the tumor volume in the treatment group was significantly lower than that in the solvent control group (P<0.05), meaning that the treatment had a significant inhibitory effect on tumor growth.

[0439] During the study period, tumor growth was strictly monitored in all animals, and tumor measurements were taken twice a week. The results were recorded, and the tumor growth suppression rate was calculated. The results of the tumor growth suppression rate study are shown in Table 32, the tumor volume change curve is shown in Figure 14, and the relative tumor volume change curve is shown in Figure 15.

[0440] [Table 32]

[0441] The results of the trial for the tumor volume ratio (T / C) between the treatment group and the control group are shown in the table below. [Table 33]

[0442] Based on the above data, the tumor growth inhibition rate (based on tumor weight and tumor volume) at the end of the study (D32) was less than 60% for epirubicin administered via tail vein, and the T / C ratio was greater than 40%. On the other hand, for Apt AS1411-4 Bio-DNA-epirubicin targeted drugs administered via tail vein at high and medium doses, the tumor growth inhibition rate (based on tumor weight and tumor volume) at the end of the study (D32 or D25) was greater than 60%, and the T / C ratio was less than 40% for both. Furthermore, even with Apt AS1411-4 Bio-DNA-epirubicin targeted drugs administered via tail vein at low doses, the tumor growth inhibition rate at the end of the study (D32) was significantly higher compared to epirubicin administered at the same dose. These results suggest that Apt AS1411-4*Bio-DNA-epirubicin targeted therapy administered via tail vein has a significant growth-inhibiting effect against lung cancer A549.

[0443] V. In Vivo Pharmacodynamic Test of Epirubicin Targeted Drug Apt AS1411-Apt EGFR-Apt A15-DNA-Epirubicin Prepared in Example 6 and Epirubicin Targeted Drug Apt AS1411-Apt EGFR-Apt A15-AmiR-21-DNA-Epirubicin Prepared in Example 20 in A549 Mouse Lung Cancer Model Experimental Animals Species / Strain: BALB / cAnSlacNifdc-nu Gender: Male Body Weight: 17 - 21 g Experimental Method 1. Inoculation and Grouping: BALB / cAnSlacNifdc-nu mice (male, body weight 17.0 - 21.0 g) were used for the A549 mouse lung cancer model. Under aseptic conditions, A549 tumor tissue was excised, cut into tumor pieces with a size of 2.0 mm × 2.0 mm × 2.0 mm, and evenly inoculated subcutaneously on the left axillary and dorsal part of the nude mice. The inoculation day was designated as D0. When the tumor volume reached approximately 150.0 mm 3 (at D7), random grouping was performed based on tumor volume, and divided into 4 groups: A (solvent control), B, C, D, with 8 mice in each group. Intravenous injection was started in all groups on the day of grouping. Groups A, B, C, D1 were administered once a week for a total of 3 times, and D2 was administered twice a week for a total of 6 times. The details of animal inoculation and grouping are shown in the following table.

[0444] 2. Administration Scheme:

Table 34

[0445] Examination Index 1. General Clinical Observation: During the test period, the clinical status of the mice was observed daily, and the rupture and ulceration status of tumor nodules, the mental state, hair coat, behavior, food intake, and excretion status of the animals were recorded.

[0446] 2. Body Weight: The body weight of the mice was weighed twice a week, the data was recorded, and the changes were analyzed.

[0447] 3. Tumor volume (V): Tumor volume was measured twice a week using calipers, and the longest and shortest diameters of the tumor were recorded. The formula for calculating volume was as follows: Volume (V) = 0.5 × Longest diameter (a) × Shortest diameter (b) 2 3. Relative tumor volume (RTV): The RTV was calculated using the following formula: RTV = Vt / V0 Here, V0 is the tumor volume measured at the time of group administration (i.e., D0), and Vt is the tumor volume at each measurement time point.

[0448] 5. Tumor growth suppression rate: The tumor growth inhibition rate (%) was calculated using the following formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100% T / C = Treatment group RTV / Control group RTV Effectiveness was determined when the tumor growth inhibition rate was 60% or higher, and statistical testing showed that the tumor volume in the treatment group was significantly lower than that in the solvent control group (P<0.05), indicating a significant inhibitory effect on tumor growth.

[0449] During the study period, tumor growth was closely monitored in all animals, and tumor measurements were taken twice a week. The results were recorded, and the tumor growth suppression rate was calculated. The results of the tumor growth suppression rate study are shown in Table 35, the tumor volume change curve is shown in Figure 16, and the relative tumor volume change curve is shown in Figure 17.

[0450] [Table 35]

[0451] The results of the trial for the tumor volume ratio (T / C) between the treatment group and the control group are shown in the table below. [Table 36]

[0452] Based on the above data, the epirubicin active pharmaceutical ingredient administered via tail vein showed a tumor growth inhibition rate (based on tumor volume) of less than 60% at the end of the study (D29), and a T / C ratio greater than 40%. On the other hand, the intravenously administered Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin targeted drugs all showed tumor growth inhibition rates (based on tumor volume) exceeding 60% at the end of the study (D29), and T / C ratios were all less than 40%. These results indicate that the Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin targeted drugs administered via tail vein have a significant growth inhibitory effect against lung cancer A549.

[0453] VI. In vivo efficacy study of the epirubicin-targeted drug Apt AS1411-Apt EGFR-AptA15-DNA-epirubicin prepared in Example 6 in a HepG2 mouse liver cancer model: Laboratory animals Species and strain: BALB / cAnSlacNifdc-nu Gender: Male Weight: 16~18 g Experimental method 1. Inoculation and Grouping: BALB / cAnSlacNifdc-nu naked mice (male, body weight 16.0-18.0 g) were used as the mouse liver cancer HepG2 model. HepG2 tumor tissue was excised under sterile conditions, cut into 2.0 mm × 2.0 mm × 2.0 mm tumor fragments, and uniformly inoculated subcutaneously into the left axillary dorsal region of the naked mice. Day of inoculation was designated as D0. The tumor volume was approximately 250.0 mm³. 3 At the point when the tumor volume was reached (D7), the animals were randomly divided into three groups based on tumor volume: A (solvent control), B, and C, with 8 animals in each group. Intravenous administration of the drug was started in all groups on the day of group division, and administered once a week for a total of three doses.

[0454] 2. Administration scheme: [Table 37]

[0455] Test indicators 1. General clinical observation: During the study period, the clinical condition of the mice was observed daily, and the rupture and ulceration status of tumor nodules, the animals' mental state, coat condition, behavior, feeding, and excretion were recorded.

[0456] 2. Weight: We weighed the mice twice a week, recorded the data, and analyzed the changes.

[0457] 3. Tumor volume (V): Tumor volume was measured twice a week using calipers, and the longest and shortest diameters of the tumor were recorded. The formula for calculating volume was as follows: Volume (V) = 0.5 × Longest diameter (a) × Shortest diameter (b) 2 4. Relative tumor volume (RTV) The RTV was calculated using the following formula: RTV = Vt / V0 Here, V0 is the tumor volume measured at the time of group administration (i.e., D0), and Vt is the tumor volume at each measurement time point.

[0458] 5. Tumor growth suppression rate: The tumor growth inhibition rate (%) was calculated using the following formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100% T / C = Treatment group RTV / Control group RTV Effectiveness was determined when the tumor growth inhibition rate was 60% or higher, and statistical testing showed that the tumor volume in the treatment group was significantly lower than that in the solvent control group (P<0.05), indicating a significant inhibitory effect on tumor growth.

[0459] 6. Tumor Weight: At the end of the study (D23), the remaining animals were euthanized by cervical dislocation, the tumors of the mice were dissected and weighed, and the tumors were photographed and recorded. The tumor weight reduction rate (IRTW) was calculated, and IRTW (%) > 60% was used as a reference indicator of effectiveness. The calculation formula is as follows: Tumor weight reduction rate (IRTW) = (W solvent control group - W treatment group) / W solvent control group × 100. Here, W represents the tumor weight.

[0460] During the study period, tumor growth was closely monitored in all animals, and tumor measurements were taken twice a week. The results were recorded, and the tumor growth suppression rate was calculated. The results of the tumor growth suppression rate study are shown in Table 38, the tumor volume change curve is shown in Figure 18, and the relative tumor volume change curve is shown in Figure 19.

[0461] [Table 38]

[0462] The results of the trial for the tumor volume ratio (T / C) between the treatment group and the control group are shown in the table below. [Table 39]

[0463] All animals were euthanized by cervical dislocation 23 days after HepG2 vaccination, and the tumors were dissected and measured. The tumor weight reduction rate (IRTW%) is shown in the table below. [Table 40]

[0464] Based on the above data, the epirubicin active pharmaceutical ingredient administered via tail vein showed a tumor growth inhibition rate (based on tumor weight and tumor volume) of less than 60% at the end of the study (D23), and a T / C ratio greater than 40%. On the other hand, the intravenously administered Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin targeted drug showed a tumor growth inhibition rate (based on tumor volume) of more than 60% at the end of the study (D23), and the T / C ratio was less than 40% in both cases. These results indicate that the Apt AS1411-Apt EGFR-Apt A15-DNA-epirubicin targeted drug administered via tail vein has a significant growth inhibitory effect on liver cancer HepG2.

[0465] VII. In Vivo Pharmacodynamic Study of Epirubicin Targeted Drug Apt TfRA3-Apt AS1411-Apt A15-DNA-Epirubicin Prepared in Example 9 in U251 Glioma Mouse Model Cell Information U251-RFP: A human glioma cell line expressing red fluorescent protein. Supplier: Antaikang Biotechnology (Beijing) Co., Ltd.

[0466] Experimental Animals Species and Strain: Balb / c nude (immunodeficient mice) Age: 4 weeks old Gender: Female Body Weight: 15 - 18 g Animal Breeding Location: In the SPF-grade barrier facility environment of Antaikang Biotechnology (Beijing) Co., Ltd., using a laminar flow cabinet and breeding according to the standards of the animal facility. Three animals were placed in each cage. The animal room had good lighting, good ventilation and air conditioning equipment, and the room temperature was maintained at 18 - 25 °C and the relative humidity at 50 - 70%.

[0467] Experimental Methods 1. Inoculation and Grouping: Thirty-two 4-week-old female BALB / c-nu mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Experimental Animal Quality Certificate Number: SCXK Jing 2016-0006). Using the surgical orthotopic transplantation method, U251-RFP tumor tissue derived from subcutaneous tissue of syngeneic mice was collected. After excising necrotic tissue under a dissecting microscope, the tumor tissue was cut into 1 mm 3The tumor tissue was cut into large, uniform pieces. Under heteroflurane anesthesia and sterile surgical conditions, the cut tumor tissue pieces were implanted into the skulls of test mice using surgical orthotopic implantation. The skull was repositioned using the inlay technique, and the mouse scalp was intermittently sutured with 6-0 surgical sutures (ETHICON.INC) to construct a mouse orthotopic model of human glioma (U251-RFP) expressing red fluorescent protein. Day of inoculation was designated as D0. On D3, non-invasive real-time fluorescence imaging was performed on the brain tumors of all mice using a FluorVivo Model-100 (INDEC BioSystems, CA, USA) fluorescence imaging device. Subsequently, the mice were randomly divided into groups, and administration to each group was started on the day of group division.

[0468] 2. Administration scheme: [Table 41]

[0469] The dosage for all test drugs was 200 μL / 20 g mice, and the actual dose was calculated based on the mouse's body weight at the time of administration.

[0470] Test indicators a) Tumor area: The fluorescence area was measured twice a week using a fluorescence imaging device. The tumor growth inhibition rate (%) was calculated using the formula: Tumor growth inhibition rate (%) = (1 - T / C) × 100%, where T / C = Treatment group RTV / Control group RTV. Here, fluorescence area was used as volume data.

[0471] b) Post-administration response in animals: Tumor volume was measured and mouse body weight was weighed simultaneously, twice a week. The relationship between changes in mouse body weight and the timing of administration was recorded, and the animals' activity level, general condition such as feeding, and survival status were observed during the administration period.

[0472] c) Imaging of tumors and organs: After administration, the mice were euthanized, and the brain, heart, liver, spleen, lungs, and kidneys were removed. These organs were placed alongside the original tumors of the corresponding groups, and first, photographs were taken, followed by imaging using tumor autofluorescence wavelength and epirubicin fluorescence wavelength, respectively.

[0473] d) Imaging of the tumor body: The primary tumor was isolated, its weight was measured, and photographs were taken (photography using a camera and imaging at tumor autofluorescence wavelength and epirubicin fluorescence wavelength).

[0474] For imaging of tumors and organs, one mouse was selected from each group, and in addition to the already collected tumor fragments, organs such as the heart, liver, spleen, lungs, kidneys, and large intestine were collected and imaged using tumor autofluorescence wavelength and epirubicin fluorescence wavelength, respectively. After the experiment, the carcasses and organs of the mice were stored in a -20°C freezer before being discarded.

[0475] The tumor weight reduction rate (IRTW) was calculated, and an IRTW (%) > 60% was used as a reference indicator of effectiveness. The calculation formula is as follows: Tumor weight reduction rate (IRTW) = (W solvent control group - W treatment group) / W solvent control group × 100. Here, W represents the tumor weight.

[0476] result During the administration period, the test animals maintained good general conditions, including activity and feeding. Around day 10 after administration, the third group of mice began to lose body weight and their spontaneous activity decreased. Drug toxicity was considered a possible cause.

[0477] During the study period, tumor growth was closely monitored in all animals, and tumor measurements were taken twice a week. The results were recorded, and the tumor growth suppression rate was calculated. The results of the tumor growth suppression rate study are shown in the table below.

[0478] [Table 42]

[0479] The results for tumor weight and tumor weight reduction rate (IRTW%) are shown in Table 43, and tumor images for groups 1, 2, and 4 are shown in Figure 20.

[0480] [Table 43]

[0481] As is clear from the table above, compared to the epirubicin active pharmaceutical ingredient, the epirubicin-targeted drug Apt TfRA3-Apt AS1411-Apt A15-DNA-epirubicin provided by the present invention showed a more significant inhibitory effect on the growth of U251-RFP tumors. Furthermore, compared to the positive control group temozolomide, the present invention was able to achieve a higher tumor growth inhibition rate at a lower dose.

[0482] 8. In vivo efficacy study of the epirubicin-targeted drug Apt TfRA4-Apt AS1411-Apt A15-DNA-epirubicin (1) prepared in Example 8 in a GL261-Luc glioma mouse model: cell culture GL261-Luc cells were purchased from Ningbo Mingzhou Biotechnology Co., Ltd. Species identification confirmed that these cells are mouse cell lines, and no contamination from rat or human-derived cells was detected. Cell culture was performed in DMEM medium containing 10% fetal bovine serum, 1% glutamine, and 1% HEPES, and maintained in a cell culture incubator at 37°C and 5% CO2. Cells were subcultured every 3-4 days when they had grown to cover the entire surface of the flask, and tumor cells in the logarithmic growth phase were used for inoculation into endogenous tumors.

[0483] Laboratory animals Species and strain: C57BL / 6 Grade: SPF Age: 6-8 weeks Gender: Female Weight: 18~22g Animal Rearing Location: Experimental animals were reared at Youji (Tianjin) Pharmaceutical Technology Co., Ltd. All animals were reared in an IVC system in a barrier rearing room. After arrival, all animals underwent quarantine for at least three days, and were incorporated into the formal trial only after all animals passed quarantine. Throughout the trial period, the temperature of the rearing room was set to 20.5-24.5°C, the humidity to 40-70%, and the light-dark cycle to 12 hours light-12 hours dark. Each rearing cage housed 3-5 animals, and sufficient water and feed were ensured throughout the trial period.

[0484] Experimental method 1. Inoculation and group division: GL261-Luc cells were resuspended in PBS to a concentration of 1 × 10⁹ + M / mL, and then administered via intracerebral injection. The inoculation dose was 5 μL / animal. Group division and administration were carried out on day 7 after tumor cell inoculation, resulting in two groups of 6 animals each. The specific administration scheme is as follows.

[0485] 2. Administration scheme: [Table 44]

[0486] Test indicators (1) After dividing the mice into groups, imaging and BLI values ​​were performed on each group of mice using an in vivo imaging device for small animals, and the body weight of the mice was weighed twice a week. The relationship between changes in mouse body weight and the timing of administration was recorded, and the survival status and health status of the animals during the administration period, i.e., general conditions such as activity and feeding, were observed.

[0487] (2) Each group of mice was observed daily, and the survival status of tumor-bearing mice was recorded. If a mouse developed the disease and became critically ill, it was euthanized, and its survival period was recorded. The observation period was a maximum of 138 days after tumor cell inoculation. At the end of the experiment, the median survival time (MST) of tumor-bearing mice in each group was calculated, and the survival extension rate (ILS%) of tumor-bearing mice in the treatment group was calculated according to the following formula.

[0488] The survival rate extension rate (ILS%) in cancer-bearing mice in the treatment group = (median survival days in the treatment group / median survival days in the solvent control group - 1) × 100% (3) The median survival time and survival extension rate of mice in each group were calculated and statistically analyzed. A p<0.05 result was considered statistically significant. BLI value change curves were created for each group of mice and statistically analyzed, with a p<0.05 result being considered statistically significant.

[0489] (4) Upon death of the mice, brain tissue was fixed with paraformaldehyde, embedded in paraffin, and then pathological sections were prepared, stained with HE, and observed and photographed.

[0490] (5) For mice that were still alive at the end of the observation period, blood was drawn from the eyes after anesthesia,...

Claims

1. A targeted chemical agent comprising a targeted nucleic acid carrier and a small molecule compound supported on the targeted nucleic acid carrier, wherein the targeted nucleic acid carrier comprises a nucleic acid carrier and a target molecule bound to the nucleic acid carrier, and the nucleic acid carrier comprises a DNA carrier or an RNA carrier or sequences A, B, and C that form a self-assembling structure. The aforementioned sequence A is SEQ ID NO. 1: 5'-ACGAGCGTTCCG-3'; The aforementioned sequence B is SEQ ID NO. 2: 5'-CGGTTCGCCG-3'; The aforementioned sequence C is SEQ ID NO. 3: 5'-CGGCCATAGCCGT-3'; or The aforementioned sequence A is SEQ ID NO. 4: 5'-ACGAGCGUUCCG-3'; The aforementioned sequence B is SEQ ID NO. 5: 5'-CGGUUCGCCG-3'; The aforementioned sequence C is SEQ ID NO. 6: 5'-CGGCCAUAGCCGU-3'; Alternatively, the sequence A, sequence B, and sequence C may include at least one base substitution, insertion, or deletion, which may be arbitrarily selected. A targeted chemical agent characterized by the following features.

2. A targeted chemical agent according to claim 1, wherein extended base segments are independently ligated to the 5' and 3' ends of sequences A, B, and C, respectively, and the extended base segments contain 0 to 14 bases; preferably, the nucleic acid carrier contains the following sequences that form a self-assembling structure: The first set of arrangements: Sequence A: SEQ ID NO. 7: 5'-GCGGCGCCCACGAGCGTTCCGGGAGC-3'; Sequence B: SEQ ID NO. 8: 5'-GCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 9: 5'-GGCGGCAGGCGGCCATAGCCGTGGGCGCCGC-3'; or The second set of arrangements: Sequence A: SEQ ID NO. 10: 5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 11: 5'-GCTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C: SEQ ID NO. 12: 5'-CGCGGCTCGCGGCCATAGCCGTGGGCGCCGC-3'; or The third set of arrangements: Sequence A: SEQ ID NO. 10: 5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 13: 5'-GCTCCTCTCCCGGTTCGCCGCCAGCCGCGG-3'; Sequence C: SEQ ID NO. 14: 5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The fourth set of arrangements: Sequence A: SEQ ID NO. 15: 5'-GCGGCGCCACCGAGCGTTCCGGGAGAGGCC-3'; Sequence B: SEQ ID NO. 16: 5'-GGCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 14: 5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The fifth set of arrangements: Sequence A: SEQ ID NO. 17: 5'-GCGGCGCCCACGAGCGUUCCGGGAGAGGCC-3'; Sequence A: SEQ ID NO. 18: 5'-GGCCUCUCCCGGUUCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 19: 5'-GGCGGCUGGCGGCCAUAGCCGUGGGCGCCGC-3'; or The sixth set of arrangements: Sequence A: SEQ ID NO. 20: 5'- GCGGCGCCCACGAGCGTTCCGGGAGAGGAGGCC-3'; Sequence B: SEQ ID NO. 21: 5'- GGCCTCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 14: 5'- GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The seventh set of arrangements: Sequence A: SEQ ID NO. 10: 5'-GCGGCGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 22: 5'-GCTCCTCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 14: 5'-GGCGGCTGGCGGCCATAGCCGTGGGCGCCGC-3'; or The eighth set of arrangements: Sequence A: SEQ ID NO. 23: 5'-GCGACGCCCACGAGCGTTCCGGGAGAGGAG-3'; Sequence B: SEQ ID NO. 24: 5'- CTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C: SEQ ID NO. 25: 5'- CGCGGCACGCGGCCATAGCCGTGGGCGTCGC-3'; or The 9th set of arrangements: Sequence A: SEQ ID NO. 26: 5'-GCGACGCCCACGAGCGTTCCGGGAGAGGAGC-3'; Sequence B: SEQ ID NO. 11: 5'-GCTCCTCTCCCGGTTCGCCGCGAGCCGCG-3'; Sequence C: SEQ ID NO. 27: 5'- CGCGGCTCGCGGCCATAGCCGTGGGCGTCGC-3'; or The 10th set of arrangements: Sequence A: SEQ ID NO. 28: 5'- GACGCCCACGAGCGTTCCGGGAGAGG-3'; Sequence B: SEQ ID NO. 29: 5'- CCTCTCCCGGTTCGCCGCGAGCCT-3'; Sequence C: SEQ ID NO. 30: 5'-GGCTCGCGGCCATAGCCGTGGGCGTCTGCTGCTGCTGCTG -3'; or The 11th set of arrangements: Sequence A: SEQ ID NO. 31: 5'- GCCCACGAGCGTTCCGGGAGA-3'; Sequence B: SEQ ID NO. 32: 5'- TCTCCCGGTTCGCCGCCAGCCGCC-3'; Sequence C: SEQ ID NO. 33: 5'- GGCGGCTGGCGGCCATAGCCGTGGGC-3' A targeted chemical agent characterized by the following features.

3. A targeted chemical agent according to claim 2, wherein the targeted chemical agent further comprises an oligonucleotide effector molecule and / or an immunostimulant supported on the targeted nucleic acid carrier, Preferably, the oligonucleotide effector molecule comprises one or more of ASOs, siRNAs, miRNAs, and nucleic acid aptamers; More preferably, the siRNA comprises one or more of ASAP1, ATAD2, CD24, CD47, EGFR, HBV, HSP, HS70, PD-L1, PAPP-1, Survivin, TAP, TIM-3, TGF-β1, and VEGF-C; even more preferably, the siRNA comprises one or more of ASAP1, CD47, PD-L1, and TGF-β1; More preferably, the miRNA comprises one or more of A-miR21, A-miR-10a, A-miR-30c, miR-34, miR-542, miR-126-3p, and miR-122; even more preferably, the miRNA is A-miR21 or; More preferably, the nucleic acid aptamer includes a nucleic acid aptamer in DNA form and / or a nucleic acid aptamer in RNA form, and preferably the nucleic acid aptamer is A1, A15, AS1411, AFP, ATP, Act-12c, A18, BAF7-1, C-Met-SL1, CH6, CA2, C12, CRAC Orail, CEA, CEA-18, CEA-T84, CSC1, CSC13, CD40, CD16a, CD19, CD3-4, CD44, CD12 (HDLBP), CD20, CD24, CD33, CD38, CD105, CD117, CD63, CD123, EGFR, EpCAM, EcR, FAP, GPC-1, GSK836, GPC3 (APS63-1), Her2, Her3, HMGA2, H2, HbsAg, IFN-y (B4), IL- 4Ra, IL-17, LZH8, MUC1, M5, M7, M1, N5, NG-Dua, NKG2D (20-N-15), NSE, Np-A15, Np-A48, Np-A58, Np-A61, OX40, PSMA, PDGFR β, PDGF, PD-L1, PD-1, PTK-7, ProGRP-48, SF, TBA15, TBA29, TfRA4, TfRA3, TTA1, TLS9a, TGF-βII (S58), TNF-a, TNF, T1, VEGF , VCAM-1, VCAM-12d, CH6, PL-45, EP66, AGC, Karpas299, SW620, MDA-MB-231, MCF-7, PC-3, BCMA, CTLA-4, CCL1, CD4-3, CD28, FGF2 (F2), FGF2, FGF5, LAG-3, MRP1, TIM3, TIMC-11, VEGF165, 4-1BBB - one or more of these are selected; more preferably, the nucleic acid aptamers include one or more of A1, A15, AS1411, C12, CD40L, EGFR, GPC-1, IL-4Ra, MUC1, OX40, PD-L1, TTA1, TfRA3, TfRA4; Preferably, the immunostimulant comprises one or more of the following: CPG2006, CPG1826, CPG2216, CPG2395, CPG-ODNT7, and CPG-ODN-PCIF1; more preferably, the immunostimulant is CPG2006. A targeted chemical agent characterized by the following features.

4. A targeted chemical agent according to claim 3, The sequence structure of the sense strand and antisense strand of each siRNA from the 5' end to the 3' end is as follows: ASAP1: Sense strand: SEQ ID NO. 34: 5'-UGAUAUUAUGGAAGCAAAUUU-3', Antisense strand: SEQ ID NO. 35: 5'-AUUUGCUUCCAUAAUAUCAUU-3'; or Sense strand: SEQ ID NO. 36: 5'-UUAGGUUUGGGGUUGGAUCUU-3, Antisense strand: SEQ ID NO. 37: 5'-GAUCCAACCCCAAACCUAAUU-3'; ATAD2: Sense strand: SEQ ID NO. 38: 5'-AAUCCUACAACUUCGACGCUU -3', Antisense strand: SEQ ID NO. 39: 5'-GCGUCGAAGUUGUAGGAUUUU-3'; CD24: Sense strand: SEQ ID NO. 40: 5'-UGUUUACAUUGUUGAGGUAUU-3', Antisense strand: SEQ ID NO. 41: 5'-UACCUCAACAAUGUAAACUUU-3'; CD47: Sense strand: SEQ ID NO. 42: 5'-GGUGAUUACCCAGAGAUAUTT-3', Antisense strand: SEQ ID NO. 43: 5'-AUAUCUCUGGGUAAUCACCTT-3'; or Sense strand: SEQ ID NO. 44: 5'-UGGUGAAAGAGGUCAUUCCUU-3', Antisense strand: SEQ ID NO. 45: 5'-GGAAUGACCUCUUUCACCAUU-3'; or Sense strand: SEQ ID NO. 46: 5'-GGAAUGACCUCUUUCACCATT-3', Antisense strand: SEQ ID NO. 47: 5'-UGGUGAAAGAGGUCAUUCCTT-3'; or Sense strand: SEQ ID NO. 48: 5'-GGUGAUUACCCAGAGAUAUUU-3', Antisense strand: SEQ ID NO. 49:5'-AUAUCUCUGGGUAAUCACCUU-3'; Preferably, the base modifications in CD47 are as follows: adjacent bonds in the range of the 2nd to 4th bases from the 5' end of the sense strand are phosphorothioate (PS) bonds, adjacent bonds in the range of the 1st to 3rd bases from the 3' end of the sense strand are PS bonds, adjacent bonds in the range of the 1st to 2nd bases from the 5' end of the antisense strand are PS bonds, and adjacent bonds in the range of the 1st to 3rd bases from the 3' end of the antisense strand are PS bonds; EGFR: Sense strand: SEQ ID NO. 50: 5'-GGCUGGUUAUGUCCUCAUUUU-3', Antisense strand: SEQ ID NO. 51: 5'-AAUGAGGACAUAACCAGCCUU-3'; or Sense strand: SEQ ID NO. 52: 5'-UUAGAUAAGACUGCUAAGGUU-3', Antisense strand: SEQ ID NO. 53: 5'-UUAGAUAAGACUGCUAAGGUU-3'; or Sense strand: SEQ ID NO. 54: 5'-UGCCUUAGCAGUCUUAUCUAAUU-3', Antisense strand: SEQ ID NO. 55: 5'-UUAGAUAAGACUGCUAAGGCAUU-3'; or Sense strand: SEQ ID NO. 78: 5'-UGCCUUAGCAGUCUUAUCUAAUUUU-3' Antisense chain: SEQ ID NO. 79: 5'- AAUUAGAUAAGACUGCUAAGGCAUU-3'; HBV: Sense strand: SEQ ID NO. 56: 5'-GGACUUCUCUCAAUUUUCUUU-3', Antisense strand: SEQ ID NO. 57: 5'-AGAAAAUUGAGAGAAGUCCUU-3'; Preferably, in the HBV, 2'-F (2'-fluoro) modifications are present on C and U; HSP: Sense strand: SEQ ID NO. 58: 5'-CGCAGAACACCGUGUUCGAUU-3', Antisense strand: SEQ ID NO. 59: 5'-UCGAACACGGUGUUCUGCGUU-3'; Preferably, in the HSP, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the sense strand are PS bonds, and adjacent bonds in the range of the 1st to 3rd bases from the 3' end of the antisense strand are PS bonds; HS70: Sense strand: SEQ ID NO. 60: 5'-GGCCAACAAGAUCACCAUCUU-3', Antisense strand: SEQ ID NO. 61: 5'-GAUGGUGAUCUUGUUGGCCUU-3'; Preferably, in HS70, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the antisense strand are PS bonds, and adjacent bonds in the range of the 1st to 3rd bases from the 3' end are PS bonds; PD-L1: Sense strand: SEQ ID NO. 62: 5'-CCAGCACACUGAGAAUCAAUU-3', Antisense strand: SEQ ID NO. 63: 5'-UUGAUUCUCAGUGUGCUGGUU-3'; or Sense strand: SEQ ID NO. 64: 5'-AGACGUAAGCAGUGUUGAATT-3', Antisense strand: SEQ ID NO. 65: 5'-UUCAACACUGCUUACGUCUTT-3'; PAPP-1: Sense strand: SEQ ID NO. 66: 5'-GAGGAAGGUAUCAACAAAUTT-3', Antisense strand: SEQ ID NO. 67: 5'-AUUUGUUGAUACCUUCCUCTT-3'; Survivin: Sense strand: SEQ ID NO. 70: 5'-GCAGGUUCCUUAUCUGUCACAUU-3', Antisense strand: SEQ ID NO. 71: 5'-UGUGACAGAUAAGGAACCUGCAGUU-3'; Alternatively, sense strand: SEQ ID NO. 72: 5'-GGAAUUGGAAGGCUGGGAACCUU-3', antisense strand: SEQ ID NO. 73: 5'-GGUUCCCAGCCUUCCAAUUCCUU-3'; preferably, adjacent bonds in the range from the 5' end of the sense strand to the 1st to 3rd bases are PS bonds, adjacent bonds in the range from the 5' end of the antisense strand to the 1st to 3rd bases are PS bonds, and furthermore, PS modification is applied to the 1st to 2nd bases from the 3' end of the antisense strand; Alternatively, sense strand: SEQ ID NO. 74: 5'-UGCAGGUUCCUUAUCUGUCATT-3', antisense strand: SEQ ID NO. 75: 5'-UGACAGAUAAGGAACCUGCTT-3'; Alternatively, sense strand: SEQ ID NO. 76: 5'-CUGCAGGUUCCUUAUCUGUCACAUU-3', antisense strand: SEQ ID NO. 77: 5'-UGUGACAGAUAAGGAACCUGCAGUU-3'; preferably, adjacent bonds in the range from the 5' end to the 1st to 3rd bases of the sense strand are PS bonds, and PS modifications are applied to the 1st to 3rd bases from the 3' end, and adjacent bonds in the range from the 5' end to the 1st to 3rd bases of the antisense strand are PS bonds, and PS modifications are applied to the 1st to 3rd bases from the 3' end; TAP: Sense strand: SEQ ID NO. 80: 5'-GCUGCACACGGUUCAGAAUUU-3', Antisense strand: SEQ ID NO. 81: 5'-AUUCUGAACCGUGUGCAGCUU-3'; Preferably, the adjacent bonds in the range from the 5' end of the sense strand to the 1st to 3rd bases are phosphorothioate (PS) bonds, the adjacent bonds in the range from the 5' end of the antisense strand to the 1st to 3rd bases are PS bonds, and further, the adjacent bonds in the range from the 3' end of the antisense strand to the 1st to 3rd bases are PS bonds; or, Sense strand: SEQ ID NO. 82: 5'-CAGGAUGAGUUACUUGAAAUU -3', Antisense strand: SEQ ID NO. 83: 5'- UUUCAAGUAACUCAUCCUGUU -3'; TIM-3: Sense strand: SEQ ID NO. 86: 5'-GUGCUCAGGACUGAUGAAATT-3', Antisense strand: SEQ ID NO. 87: 5'-UUUCAUCAGUCCUGAGCACTT-3'; TGF-β1: Sense strand: SEQ ID NO. 88: 5'-GUCAACUGUGGAGCAACACUU-3', Antisense strand: SEQ ID NO. 89: 5'-GUGUUGCUCCACAGUUGACUU-3'; or Sense strand: SEQ ID NO. 90: 5'-GCAACAACGCCAUCUAUGATT-3', Antisense strand: SEQ ID NO. 91: 5'-UCAUAGAUGGCGUUGUUGCTT-3'; VEGF-C: Sense strand: SEQ ID NO. 92: 5'-GCAAGACGUUGUUUGAAAUUAUU-3', Antisense strand: SEQ ID NO. 93: 5'-UAAUUUCAAACAACGUCUUGCUU-3'; Preferably, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the sense strand are PS bonds, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the antisense strand are PS bonds, and further adjacent bonds in the range of the 1st to 3rd bases from the 3' end of the antisense strand are PS bonds; or, Sense strand: SEQ ID NO. 84: 5'-CAGCAAGACGUUGUUUGAAAUUAUU -3', Antisense strand: SEQ ID NO. 85: 5'- UAAUUUCAAACAACGUCUUGCUGUU -3'; or, Sense strand: SEQ ID NO. 94: 5'-CAGGAUGGUAAAGACUACAUU-3', Antisense strand: SEQ ID NO. 95: 5'-UGUAGUCUUUACCAUCCUGUU-3; Preferably, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the sense strand are PS bonds, adjacent bonds in the range of the 1st to 3rd bases from the 5' end of the antisense strand are PS bonds, and further adjacent bonds in the range of the 1st to 3rd bases from the 3' end of the antisense strand are PS bonds; Preferably, the miRNA includes one or more of the following: miR-34, miR-542, miR-126-3p, and miR-122; Preferably, the ASO includes one or more of A-miR21, A-miR-10a, A-miR-30c, and A-miR-1306. The sequence structure of each miRNA and ASO from the 5' end to the 3' end is as follows: A-miR21: 5'-GATAAGCT-3'; preferably, each base is modified with locked nucleic acid (LNA); or, 5'-GAUAAGCU-3'; preferably, each base is modified with LNA; or, SEQ ID NO. 96: 5'-GTCAACATCAGTCTGATAAGCTA-3'; or, SEQ ID NO. 97: 5'-GUCAACAUCAGUCUGAUAAGCUA-3'; or, SEQ ID NO. 98: 5'-TCAACATCAGTCTGATAAGCTA; or, 5'-GATAAGCT-3'; preferably, adjacent bonds are PS bonds; A-miR-10a:5'-ACAGGGTA-3'; preferably, each base is modified with LNA; A-miR-30c: SEQ ID NO. 99: 5'-GCTGAGAGTGTAGGATGTTTACA-3'; miR-34: 5'-TGTGACAG-3'; miR-542: 5'-TGGCAGTGT-3'; miR-126-3p:5'-UCGUACC-3'; preferably, adjacent bonds are PS bonds; miR-122: 5'-GGAAGTGT-3; AmiR-1306: SEQ ID NO. 100: 5'-CATCACCACCAGAGCCAACGTC-3'; preferably, adjacent bonds in the range of the 1st to 5th bases from the 5' end are PS bonds; The sequence structure of each nucleic acid aptamer from the 5' end to the 3' end is as follows: A1: SEQ ID NO. 101: 5'-GGTTGCATGCCGTGGGGAGGGGGGTGGGTTTTATAGCGTACTCAG-3'; A15: SEQ ID NO. 102: 5'-CCCTCCTACATAGGG-3'; or SEQ ID NO. 227: 5'-CCCUCCUACAUAGGG-3'; AS1411: SEQ ID NO. 103: 5'-GGTGGTGGTGGTTGTGGTGGTGGTGG-3'; or SEQ ID NO. 228: 5'-GGUGGUGGUGGUUGUGGUGGUGGUGG-3; or SEQ ID NO. 229: AUUCUGAACCGUGUGCAGCACCACGCUGCACACGGUUCAGAAUACACACGAGGCTATCTAGAATGTAC-3'; AFP: SEQ ID NO. 104: 5'-GGCAGGAAGACAAACAAGCTTGGCGGCGGGAAGGTGTTTAAATTCCCGGGTCTGCGTGGTCTGTGGTGCTGT-3'; or SEQ ID NO. 105: 5'-ACCTGGGGAGTATTGGGGAGGAAGG-3'; ATP: SEQ ID NO. 106: 5'-ACCTGGGGAGTATTGGGGAGGAAGG-3'; or SEQ ID NO. 107: 5'-GGGAGGACGATGCGGAGGAAGGGTAGG-3'; preferably, adjacent bonds in the range of the 1st to 4th bases from the 5' end are PS bonds; Act-12c: SEQ ID NO. 108: 5'-CGGGGAAAGTCACGGGGGGTTTCAGATGTTCTGATCGGTGTGGAG-3'; A18:SEQ ID NO. 109:5'-CCAGATAGTCCCTGG-3'; BAF7-1: SEQ ID NO. 110: 5'-GATAACGGGCACGAATTCGGAGTG-3'; C-Met-SL1: SEQ ID NO. 111: 5'-ATCAGGCTGGATGGTAGCTCGGTCGGGGTGGGTGGGTTGGCAAGTCTGAT-3'; CH6: SEQ ID NO. 112: 5'-AGTCTGTTGGACCGAATCCCGTGGACGCACCCTTTGGACG-3'; CA2: SEQ ID NO. 113: 5'-CCCACGTCTGCGCTTAGCTCCTGGGCCTGGATGGGC-3'; C12: SEQ ID NO. 114: 5'-GTGGATTGTTGTGTTCTGTTGGTTTTTGTGTTGTC-3'; preferably, adjacent bonds in the range of the 1st to 3rd bases from the 5' end are PS bonds; CRAC Orail: SEQ ID NO. 115: 5'-CCAGTAGCCATACCGGTTTGTGGATGGGGTGTATGCGAGT-3'; CEA: SEQ ID NO. 116: 5'-CTAGGATCCCCACTCACCATCTCTCAGCTTGCTTCCTAGC-3'; or SEQ ID NO. 234: 5'-CUAGGAUCCCACUCACCAUCUCUCAGCUUGCUUCCUAGC-3'; CEA-T84: SEQ ID NO. 118: 5'-TCGCGCGAGTCGTCTGGGGAACCATCGAGTTACACCGACCTTCTATGTGCGGCCCCGCATCGTCCTCCC-3'; CSC1: SEQ ID NO. 119: 5'-ACCTTGGCTGTCGTGTTGTAGGTGGTTTGCTGCGGTGGGCTCAAGAAGAAAGCGCAAAGAGGTCAGTGGTCAGAGCGT-3'; CSC13: SEQ ID NO. 120: 5'-ACCTTGGCTGTCGTGTTGTGGGGTGTCGTATCTTTCGTGTCTTATTATTTTCTAGGTGGAGGTCAGTGGTCAGAGCGT-3'; CD40: SEQ ID NO. 121: 5'-CCAACGAGTAGGCGATAGCGCGTGG-3'; or SEQ ID NO. 122: 5'-AGAGACGATGCGGCCAACGAGTAGGCGATAGCGCGTGGCAGAGCGTCGCT-3'; CD16a: SEQ ID NO. 123: 5'-CCACTGCGGGGGTCTATACGTGAGGAAGAAGTGG-3'; CD19: SEQ ID NO. 124: 5'-TGCGTGTGTAGTGTGTCGTTTCTCCTTTTTTTGGTTGCTGCTCTTAGGGATTTGGGCGG-3'; CD3-4: SEQ ID NO. 126: 5'-TCTCGGACGCGTGTGGTCGGCCGAGTGGCCCACGGTAGAAGGGTTAGAACTGCTGGTTGGTGAATCTCGCTGCCTGGCCCTAGAGTG-3'; CD44: SEQ ID NO. 127: 5'-CCAAGGCCTGCAAGGGAACCAAGGACACAG-3'; or SEQ ID NO. 128: 5'-CCAAGGCCTGCAAGGGAACCAAGGACACAG-3'; preferably, adjacent bonds in the range of 3 to 5 bases from the 5' end are PS bonds, adjacent bonds in the range of 12 to 14 bases from the 5' end are PS bonds, the bond between 1 to 2 bases from the 3' end is a PS bond, the bond between 3 to 4 bases from the 3' end is a PS bond, the bond between 5 to 6 bases from the 3' end is a PS bond, and adjacent bonds in the range of 12 to 14 bases from the 3' end are PS bonds; or, SEQ ID NO. 237: 5'-GGGAUGGAUCCAAGCUUACUGGCAUCUGGAUUUGCGCGUGCCAGAAUAAAGAGUAUAAACGUGUGAAUGGGAAGCUUCGAUAGGAAUUCGG-3'; CD12 (HDLBP): SEQ ID NO. 129: 5'-GTGGATTGTTGTGTTCTGTTGGTTTTTGTGTTGTC-3'; CD20: SEQ ID NO. 130: 5'-TGCGTGTGTAGTGTGTCTGTTTTTTATCTTCTTTTATCTACTCTTAGGGATTTGGGCGG-3'; CD24: SEQ ID NO. 131: 5'-TATGTGGGTGGGTGGGCGGTTATGCTGAGTCAGCCTTGCT-3'; or SEQ ID NO. 132: 5'-ATCCAGAGTGACGCAGCATATGTGGGTGGGTGGGCGGTTATGCTGAGTCAGCCTTGCTTGGACACGGTGGCTTAGT-3'; CD33: SEQ ID NO. 133: 5’-TACCAGTGCGATGCTCAGCACGCTTATAGGGGCTGGACAAAATTCTACCCAGCCTTT-3’; CD38: SEQ ID NO. 134: 5’-TACGTGAATCTCGTACGATACTCTGTAAGCGT-3’; CD105: SEQ ID NO. 135: 5’-GATCAGTTTTCCATGCCAGTTGGTATTCCGCGACAGTTTGATCTC-3’; CD117: SEQ ID NO. 136: 5’-GGGGCCGGGGCAAGGGGGGGGTACCGTGGTAGGAC-3’; CD63: SEQ ID NO. 137: 5’-CACCCCACCTCGCTCCCGTGACACTAATGCTA-3’; CD123: SEQ ID NO. 138: 5’-TGCGTGTGTAGTGTGTCTGGGCTACATCGATGAGCTGCCTAGGGTCCCTCTTAGGGATTTGGGCGG-3’; EGFR: SEQ ID NO. 139: 5’-GCCTTAGTAACGTGCTTTGATGTCGATTCGACAGGAGGC-3’; or, SEQ ID NO. 239: 5’-GCCUUAGUAACGUGCUUUGAUGUCGAUUCGACAGGAGGC-3’; or, SEQ ID NO. 240: 5’-UGCCGCUAUAAUGCACGGAUUUAAUCGCCGUAGAAAAGCAUGUCAAAGCCGUU-3’; EpCAM: SEQ ID NO. 140: 5’-CACTACAGAGGTTGCGTCTGTCCCACGTTGTCATGGGGGGTTGGCCTG-3’; or, SEQ ID NO. 141: 5’-GACAAACGGGGGAAGATTTGACGTCGACGAC-3’; or, SEQ ID NO. 241: 5’-GCGACUGGUUACCCGGUCG-3’; EcR: SEQ ID NO. 142: 5’-GCAGGTCCACTGCGGGGGTCTATACGTGAGGAAGAAGTGGGCAGGTC-3’; FAP: SEQ ID NO. 143: 5’-TGGGGGTTGAGGCTAAGCCGA-3’; Anti-FAP: SEQ ID NO. 144: 5'-CCGCTCGAGCTAGTCTGACAAAGAGAAACAC-3'; GPC-1: SEQ ID NO. 145: 5'-AACGGAGTGTGGCTAACTCGA-3'; GSK836: SEQ ID NO. 147: 5'-GCAGAGGTGAAGCGAAGTCG-3'; preferably, adjacent bonds in the range of the 1st to 6th bases from the 5' end are PS bonds; GPC3 (APS63-1): SEQ ID NO. 148: 5'-TAACGCTGACCTTAGCTGCATGGCTTTACATGTTCCA-3'; preferably, adjacent bonds in the range of the 1st to 4th bases from the 5' and 3' ends are PS bonds; Her2: SEQ ID NO. 149: 5'-AGCCGCGAGGGGAGGGATAGGGTAGGGCGCGGCT-3'; or SEQ ID NO. 150: 5'-GGGAGCTCAGAATAAACGCTCAAAGGGTCAAGCTGATTACACTTTGTCCACTATTGGGTCCTTCGACATGAGGCCCGGATC-3'; or SEQ ID NO. 246:5'-AGCCGCGAGGGGAGGGAUAGGGUAGGGCGCGGCU-3'; Her3: SEQ ID NO. 151: 5'-GGGAGCTCAGAATAAACGCTCAAGGCTAACAGCACGCAACGGGGGGAGTAATCGTGTCTGTTCGACATGAGGCCCGGATC-3'; or SEQ ID NO. 247: 5'-CAGCGAAAGUUGCGUAUGGGUCACAUCGCAGGCACAUGUCAUCUGGCG-3'; or SEQ ID NO. 248:5'-GAAUUCCGCGUGUGCCAGCGAAAGUUGCGUAUGGGCCACAUCGCAGGCACAUGUCAUCUGGGCGGUCCGUUCGGGAUCC-3'; HMGA2: SEQ ID NO. 152: 5'-GGAAAAAATTTTTTAAAAAACCC-3'; preferably, adjacent bonds are PS bonds; H2: SEQ ID NO. 146: 5'-GGGCCGTCGAACACGAGCATGGTGCGTGGACCTAGGATGACCTGAGTACTGTCC-3'; HBsAg: SEQ ID NO. 153: 5'-CACAGCGAACAGCGGCGGACATAATAGTGCTTACTACGAC-3'; preferably, adjacent bonds in the range of the 1st to 4th bases from the 5' end are PS bonds; IFN-y (B4): SEQ ID NO. 154: 5'-CCGCCCAAATCCCTAAGAGAAGACTGTAATGACATCAAACCAGACACACACACTACACACGCA-3'; IL-4Ra: SEQ ID NO. 155: 5'-GGAGGACGAUGCGGAAAAAGCAACAGGGUGCUCCAUGCGCAUGGAACCUGCGGCGCAGACGACUCGCUGAGGAUCCGAGA-3'; or SEQ ID NO. 156: 5'-AAAAAGCAACAGGGUGCUCCAUGCGCAUGGAACCUGCGCG-3'; IL-17: SEQ ID NO. 157: 5'-CTTGGATCACCATAGTCGCTAGTCGAGGCT-3'; or SEQ ID NO. 158: 5'-GCGGCATCCTATCACGCATTGACC-3'; LZH8: SEQ ID NO. 159: 5'-ATCCAGAGTGACGCAGCATATTAGTACGGCTTAACCCCATGGTGGACACGGTGGCTTAGT-3'; MUC1: SEQ ID NO. 160: 5'-GCAGTTGATCCTTTGGATACCCTGG-3'; or SEQ ID NO. 161: 5'-GAAGTGAAAATGACAGAACACAACA-3'; or SEQ ID NO. 162: 5'-AACCGCCCAAATCTCTAAGAGTCGGACTGCAACCTATGCTATCGTTGATGTCTGTCCAAGCAACACAGACACACTACACACACGCACA-3'; or SEQ ID NO. 163: 5'-AATGACAGAACACAACATT-3'; or SEQ ID NO. 250: 5'-GCAGUUGAUCCUUUGGAUACCCUGG-3'; M5: SEQ ID NO. 164: 5'-AGCAGCACAGAGGTCAGATGCTTGGTTCCACCGTACTGACTGTAGTAAAATCTGATCACTCCTATGCGTGCTACCGTGAA-3'; M7:SEQ ID NO. 165:5'-AGCAGCACAGAGGTCAGATGTAGTCGGTCTTCTTGTTTGAAACTGCTAATTTTGAAAAAACCTATGCGTGCTACCGTGAA-3'; M1:SEQ ID NO. 166:5'-AGCAGCACAGAGGTCAGATGATATAACCTTAATAATAAAATAAATTTTAATCTTACCTATGCGTGCTACCGTGAA-3'; N5:SEQ ID NO. 167:5'-GATTGAGTAGATAGTGGTTCTGTACGTAGTGAAAGAGTGG-3'; NG-Dua:SEQ ID NO. 168:5'-CAAGTTGCTCGTCGCGATACGTTTGGTTGGTGTGGTTGGCAGTATCGCAGGTCCAAGTTGCTCGTCGCGATACAACGGAGTGTGGCTAACTCGA-3'; NKG2D:20-N-15:SEQ ID NO. 169:5'-CAAGTTGCTCGTCGCGATACGTTTGGTTGGTGTGGTTGGCAGTATC-3'; NSE:SEQ ID NO. 170:5'-TCACACGGACCTCTCTCTACATTAATTGCGCATTTCGTT-3': Np-A15:SEQ ID NO. 171:5'-GCTGGATGTTCATGCTGGCAAAATTCCTTAGGGGCACCGTTACTTTGACACATCCAGC-3'; Np-A48:SEQ ID NO. 172:5'-GCTGGATGTCGCTTACGACAATATTCCTTAGGGGCACCGCTACATTGACACATCCAGC-3'; Np-A58:SEQ ID NO. 173:5'-GCTGGATGTCACCGGATTGTCGGACATCGGATTGTCTGAGTCATATGACACATCCAGC-3'; Np-A61:SEQ ID NO. 174:5'-GCTGGATGTTGACCTTTACAGATCGGATTCTGTGGGGCGTTAAACTGACACATCCAGC-3'; OX40: SEQ ID NO. 175: 5’-GGGAGGACGATGCGGCAGTCTGCATCGTAGGAATCGCCACCGTATACTTTCCCACCAGACGACTCGCTGAGGATCCGAGA-3’; or SEQ ID NO. 176: 5’-CAGTCTGCATCGTAGGATTAGCCACCGUATCTTTCCCAC-3’; or SEQ ID NO. 177: 5’-CCAACGAGTAGGCGATAGCGCGTGG-3’; or SEQ ID NO. 252: 5’-GGGAGGACGAUGCGGCAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCACCAGACGACUCGCUGAGGAUCCGAGA-3’; or SEQ ID NO. 253: 5’-GGGAGGACGAUGCGGCAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCACCAGACGACUCGCUG-3’; or SEQ ID NO. 254: 5’-CAGUCUGCAUCGUAGGAAUCGCCACCGUAUACUUUCCCAC-3’; or SEQ ID NO. 255: 5’-GGGAUGCGGAAAAAAGAACACUUCCGAUUAGGGCCCACCCUAACGGCCGCAGAC-3’; PSMA: SEQ ID NO. 179: 5’-GGGAGGACGATGCGGATCAGCCATGTTTACGTCACTCCT-3’; or SEQ ID NO. 180: 5’-GCGTTTTCGCTTTTGCGTTTTGGGTCATCTGCTTACGATAGCAATGCT-3’; or SEQ ID NO. 256: 5’-GGGACCGAAAAAGACCUGACUUCUAUACUAAGUCUACGUUCCC-3’; or SEQ ID NO. 257: 5’-GGGAGGACGAUGCGGAUCAGCCAUGUUUACGUCACUCCU-3’; PDGFRβ: SEQ ID NO. 181: 5’-TGTCGTGGGGCATCGAGTAAATGCAATTCGACA-3’; or SEQ ID NO. 258: 5’-UGUCGUGGGGCAUCGAGUAAAUGCAAUUCGACA-3’; PDGF:SEQ ID NO. 182:5'-CAGGCTACGGCACGTAGAGCATCACCATGATCCTG-3' PD-L1:SEQ ID NO. 183:5′-AACAACGTATAACAATGCCCACGTCACCAGAGTACTATGG-3′ and is SEQ ID NO. 184:5'-GCCCCAGTTATGCTTTCCCCCTCTGTCTCTTTG-3' and is SEQ ID NO. 185:5′-ATCGCCCGCAGCACCCATTTGTTTTTTTTTG-3′ and is SEQ ID NO. 69:ACGGGCCACATCAACTCATTGATAGACAATGCGTCCACTGCCCGT and SEQ ID NO. 186:5'-TGCCCGCACATCAACTCATTGATAGACAATGCGTCCACTCGGGCA-3' and is SEQ ID NO. 187:5'-TGCCCGCACATCAACTCATTGATAGACAATGCGTCCACTACGGGC-3' and is SEQ ID NO. 188:5'-CGGGCACACATCAACTCATTGATAGACAATGCGTCCACTGCCCGT-3' and is SEQ ID NO. 189:5′-GTTGGTCACATCAACTCATTGATAGACAATGCGTCCACTACCAAC-3′ and is SEQ ID NO. 190:5'-GGGCCACATCAACTCATTGATAGACAATGCGTCCACTGCCC -3' and SEQ ID NO. 191:5'-TGGTTGCACATCAACTCATTGATAGACAATGCGTCCACTCAACCA-3' and is SEQ ID NO. 200:5'-TACAGGTTCTGGGGGGTGGGTGGGGAACCTGTT-3' and is SEQ ID NO. 238:5'-CTACGAGACGAACTTATGCGTAATAATGACTGTCGTAG-3' PD-1:SEQ ID NO. 192:5'-GACGATAGCGGTGACGGCACAGACGGTACAGTTCCCGTCCCTGCACTACACGTATGCCGCTTCCGTCCGTCGCTC-3' is SEQ ID NO. 193:5'-GAGCGACGGACGGAAGCGGCATACGTGTAGTGCAGGGACGGGAACTGTACCGTCTGTGCCGTCACCGCTATCGTC-3' and is SEQ ID NO. 194:5'-GGATCCTAGACGCATTGACCCGCTGCCTCTACTGAGGCTGTGTCAGTGTGCGGCTCGGACTGTTGAATTC-3' and is SEQ ID NO. 195:5'-AGCGGTGACGGCACAGACGGTACAGTTCCCGTCCCTGCACTACACGTATGCCGCTGAGAGAGAGGGAGGC-3' and is SEQ ID NO. 196:5'-ACCGACAGTGAAGGACTCAGCGAACTCTCAGACTCGGTTC-3' PTK-7:SEQ ID NO. 197:5'-ATCTAACTGCTGCGCCGCCGGGAAAATACTGTACGGTTAGA-3' ProGRP-48:SEQ ID NO. 198:5'-CATGCGGAGTAGAGCGAGCCCAGATAGTCCCTGGTTATTTCCTTAGG-3' SF:SEQ ID NO. 199:5'-GATCTCTCTCTGCCCTAAGTCGCACCCGTGCTTCCCTGT-3' TBA15:SEQ ID NO. 201:5'-GGTTGGTGTGGTTGG-3' TBA29:SEQ ID NO. 202:5'-AGTCCGGTGGTAGGGCAGGTTGGGGTGACT-3' TfRA4:SEQ ID NO. 203:5'-GCGTGGTACCACGC-3' and SEQ ID NO. 259:5'-GCGUGGUACCACGC-3' TfRA3: SEQ ID NO. 204: 5'-GCGTGGTCACACGC-3'; or SEQ ID NO. 205: 5'-GCGGCGCCCACGAGCGTTCGCGTGGTCACACGCGTTCCGCCCTCCTACATAGGGCGCATAGCCGTGGGCGCCGC-3'; or SEQ ID NO. 260: 5'-GCGUGGUCACACGC-3'; TTA1: SEQ ID NO. 206: 5'-CCTGCACTTGGCTTGGATTTCAGAAGGGAGACCC-3'; or SEQ ID NO. 286: 5'-CTGCACTTGGCTTGGATTTCAGAAGGGAGACCC-3'; or SEQ ID NO. 261: 5'-CCUGCACUUGGCTTGGAUUUCAGAAGGGAGACCC-3'; TLS9a: SEQ ID NO. 207: 5'-AGTCCATTTTATTCCTGAATATTTGTAACCTCATGGAC-3'; TGF-βII (S58): SEQ ID NO. 208: 5'-ACATTGCTGCGTGATCGCCTCACATGGGTTTGTCTGGTCGATTTGGAGGTGGTGGGTGGC-3'; TNF-a: SEQ ID NO. 209: 5'-GCGGCCGATAAGGTCTTTCCAAGCGAACGAAAA-3'; TNF:SEQ ID NO. 210:5'-GCGCCACTACAGGGGAGCTGCCATTCGAATAGGTGGGCCGC-3'; T1: SEQ ID NO. 211: 5'-CGCTCGATAGATCGAGCTTCGCTCGATGTGGTGTTGTGGGGGCTTGTATTGGTCGATCACGCTCTAGAGCACTG-3'; VEGF: SEQ ID NO. 212: 5'-TGTGGGGGTGGACTGGGTGGGTACC-3'; or SEQ ID NO. 213: 5'-TGTGGGGGTGGACGGGCCGGGTAGA-3'; or SEQ ID NO. 214: 5'-GGTGGGGGTGGACGGGCCGGGTAGA-3'; or SEQ ID NO. 266: 5'-AUGCAGUUUGAGAAGUCGCGCAU-3'; preferably, the adjacent bond between the 6th and 7th bases from the 5' end is a phosphorothioate (PS) bond; VCAM-1: SEQ ID NO. 215: 5’-ATACCAGCTTATTCAATTGGACACGGCAAAGGGGTATAGCCTACCGGACCGTGAACATGGAATGGTGTGCTGCGTGGAGATAGTAAGTGCAATCT-3’; or, SEQ ID NO. 216: 5’-GGACACGGCAAAGGGGTATAGCCTACCGGACCGTGAACATGGAATGGTGTGCTGCGTGG-3’; VCAM-1 2d: SEQ ID NO. 217: 5’-AGGGAATCTTGCCTAGGGAGGGAGTAGCGAAAGGGCTCA-3’; CH6: SEQ ID NO. 218: 5’-AGTCTGTTGGACCGAATCCCGTGGACGCACCCTTTGGACG-3’; PL-45: SEQ ID NO. 219: 5’-ACTCATAGGGTTAGGGGCTGCTGGCCAGATACTCAGATGGTAGGGTTACTATGAGC-3’; EP66: SEQ ID NO. 220: 5’-AACAGAGGGACAAACGGGGGAAGATTTGACGTCGACGACA-3’; AGC: SEQ ID NO. 221: 5’-CGACCCGGCACAAACCCAGAACCATATACACGATCATTCGTCTCCTGGGCCG-3’; Karpas299: SEQ ID NO. 222: 5’-ATCCAGATGACGCAGCACCACCACCGTACAATTTTTTCATTACCTACTCGGC-3’; SW620: SEQ ID NO. 223: 5’-CCCATCAATGTTACGACCCGCTAGGGCTGCTGTGCCATCGGGTAA-3’; MDA-MB-231: SEQ ID NO. 224: 5’-AGAATTCAGTCGGACAGCGAAGTAGTTTTCCTTCTAACCTAAGAACCCGCGGCAGTTTAATGTAGA-3’; MCF-7: SEQ ID NO. 225: 5’-GCATGGGGTTTCGGCGTTTCGTCTATCTTGTTTCTGTTAGCGTCT-3’; PC-3: SEQ ID NO. 226: 5’-TGCCACTACAGCTGGTTCGGTTTGGTGACTTCGTTCTTCGTTGTGGTGCTTAGTGGC-3’; BCMA: SEQ ID NO. 230: 5’-AGUGCAAGACGUUCGCAGAUUAGCGAAAAGAGGGUCUCAUUGACUAGUAC-3’; CTLA-4: SEQ ID NO. 231: 5’-GGUGGAAGAGGGAUGGGCCGACGUGCCGCAU-3’; or, SEQ ID NO. 232: 5’-TCCCTACGGCGCTAACGATGGTGAAAATGGGCCTAGGGTGGACGGTGCCACCGTGCTACAAC-3’; CCL1: SEQ ID NO. 233: 5’-UGACUCCUCUGACAGCCUAAUUUCUCCCGAUUACCCUG-3’; CD4-3: SEQ ID NO. 235: 5’-GGGAGGACGAUGCGGUUUGGGGUUUUCCCGUGCCCCAGACGACUCGCCCGA-3’; CD28: SEQ ID NO. 236: 5’-GGGAGAGAGGAAGAGGGAUGGGGAUUAGACCAUAGGCUCCCAACCCCCCCCGGGAGAGAGGAAGAGGGAUGGGGAUUAGACCAUAGGCUCCCAACCCCCGGG-3’; FGF2 (F2): SEQ ID NO. 242: 5’-GGGAUACUAGGGCAUUAAUGUUACCAGUGUAGUCCC-3’; FGF2: SEQ ID NO. 243: 5’-GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCUCGA-3’; or, SEQ ID NO. 244: 5’-GGGAAACUAGGGCGUUAACGUGACCAGUGUUUCCC-3’; FGF5: SEQ ID NO. 245: 5’-GGGCGACCUCUCCGUACUGACCUACAGAGCGACAUACUAGUGUAUCCAGAUCGCCC-3’; LAG-3: SEQ ID NO. 249: 5’-GGGAGAGAGAUAUAAGGGCCUCCUGAUACCCGCUGCUAUCUGGACCGAUCCCAUUACCAAAUUCUCUCCC-3’; MRP1: SEQ ID NO. 251: 5’-GGGAGAAUAGUCAACAAAUCGUUUGGGGCGACUUCUCCUUCCUUUCUCCC-3’; TIM3: SEQ ID NO. 262: 5'-GGGAGAGGACCAGUA-GCCACUAUGGUGUUGGAGCUAGCGG-CAGAGCGUCGCGGUCCCUCCC-3'; TIMC-11: SEQ ID NO. 264: 5'-AAGCACACUUAGUCGCGAUUGAUACGUGCGCAGUCAU-3'; VEGF165:SEQ ID NO. 267:5'-CGGAAUCAGUGAAUGCUUA UACAUCCG-3'; 4-1BB: SEQ ID NO. 268: 5'-GGGAGAGAGGAAGAGGGAUGGGCGACCGAACGUGCCCUUCAAAGCCGUUCACUAACCAGUGGCAUAACCCAGAGGUCGAUAGUACUGGAUCCCCC-3'; The sequence structure of each of the aforementioned immunostimulants from the 5' end to the 3' end is as follows: CPG2006: SEQ ID NO. 269: 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3'; preferably, adjacent bonds are PS bonds; or, SEQ ID NO. 270: 5'-UCGUCGUUUUGUCGUUUUGUCGUU-3'; CPG1826: SEQ ID NO. 271: TCCATGACGTTCCTGACG-3'; preferably, adjacent bonds are PS bonds; CPG2216: SEQ ID NO. 272: 5'-GGGGGACGATCGTCGGGGGG-3'; CPG2395: SEQ ID NO. 273: TCGTCGTTTTCGGCGCGCGCCG-3'; CPG-ODNT7: SEQ ID NO. 274: 5'-TCGTCGTCGTCGTCGTCGTCG-3'; or SEQ ID NO. 275: 5'-TCGTCGTCGTCGTCGTCGTCGTCG-3'; CPG-ODN-PCIF1: 5'-AGCGAA-3' A targeted chemical agent characterized by the following features.

5. A targeted chemical agent according to claim 3 or claim 4, wherein if the targeted chemical agent includes the nucleic acid aptamer, the target molecule includes the nucleic acid aptamer and / or a low molecular weight target head, and if the targeted chemical agent does not include the nucleic acid aptamer, the target molecule is a low molecular weight target head, wherein the low molecular weight target head is one or more selected from the group consisting of folic acid, biotin, vitamin B12, and mannose. A targeted chemical agent characterized by the following features.

6. A targeted chemical agent according to claim 5, wherein the small molecule chemical agent is selected from anthracycline chemotherapeutic agents, pyrimidine chemotherapeutic agents, platinum chemotherapeutic agents, glutamic acid derivative chemotherapeutic agents, flavonoid chemotherapeutic agents, herbal medicines and their derivatives, folic acid chemotherapeutic agents, salicylic acid chemotherapeutic agents, or acridine chemotherapeutic agents; Preferably, the anthracycline chemotherapeutic agent is selected from doxorubicin, epirubicin, pirarubicin, daunorubicin, idarubicin, mitoxantrone, barrubicin, or their free bases or hydrochlorides; Preferably, the pyrimidine-based chemotherapeutic agent is selected from gemcitabine, 5-fluorouracil, cytarabine, or capecitabine; Preferably, the platinum-based chemotherapeutic agent is selected from cisplatin, oxaliplatin, carboplatin, nedaplatin, or lovaplatin; Preferably, the glutamate derivative chemotherapeutic agent is selected from lenalidomide, thalidomide, or pomalidomide; Preferably, the flavonoid chemotherapeutic agent is selected from flavones, flavonols, dihydroflavones, isoflavones, or chalcones; Preferably, the crude drug and its derivatives are selected from vincristine, dihydroartemisinin, paclitaxel, meitansine, docetaxel, or 10-hydroxycamptothecin; Preferably, the folic acid-based chemotherapeutic agent is methotrexate; Preferably, the salicylate-based chemotherapeutic agent is aspirin or sodium salicylate; Preferably, the acridine-based chemotherapeutic agent is selected from tacrine or aminoacridine; More preferably, the low molecular weight chemical is one or more selected from epirubicin or its free base or hydrochloride, gemcitabine, 5-fluorouracil, paclitaxel, and meitansine. A targeted chemical agent characterized by the following features.

7. The targeted chemical agent according to claim 6, wherein when the low molecular weight target head, oligonucleotide effector molecule and immunostimulant are linked to the nucleic acid carrier, they are directly linked to the terminal bases of each sequence, or linked to the terminal bases of each sequence of the nucleic acid carrier via a base linkage bridge containing 1 to 10 bases, preferably, If the small molecule chemical is the anthracycline-based chemotherapeutic agent or the acridine-based chemotherapeutic agent, the small molecule chemical is inserted between adjacent GC base pairs of the targeted nucleic acid carrier in a specific intercalation manner; If the small molecule chemical is the pyrimidine-based chemotherapeutic agent, the small molecule target head, oligonucleotide effector molecule, and immunostimulant are linked to the 5' or 3' end of each sequence of the nucleic acid carrier via a base linkage bridge containing 1 to 10 bases, and the small molecule chemical is linked to the nucleic acid carrier in the form of substituting at least some of the bases in the base linkage bridge, or as an extended base, or as a base substitution of the carrier skeleton; When the small molecule chemical is a platinum-based chemotherapeutic agent, a glutamate derivative-based chemotherapeutic agent, a flavonoid-based chemotherapeutic agent, a herbal medicine and its derivatives, a folic acid-based chemotherapeutic agent, or a salicylic acid-based chemotherapeutic agent, the small molecule chemical is bound to a nucleic acid carrier by covalent bonding via a linker; preferably, the linker is selected from long-chain primary amine compounds, more preferably a long-chain primary amine compound having C2 to C14 carbon atoms, and even more preferably 5-(bromomethyl)-6-bromohexane-1-amine. A targeted chemical agent characterized by the following features.

8. A targeted chemical agent according to claim 7, wherein the oligonucleotide effector molecule, immunostimulant, and low molecular weight target head are ligated to one or more sequences in the nucleic acid carrier by chain extension or chain complementarity formation, and preferably the targeted chemical agent is any of the following: A) The targeted chemical agent comprises a DNA carrier, a small molecule chemical agent, and a nucleic acid aptamer EGFR linked to the DNA carrier, and optionally biotin, preferably the small molecule chemical agent being epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a first sequence, and the nucleic acid aptamer EGFR is linked to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer EGFR is linked to the 5' end of sequence B by a base linkage bridge TTTTT; if the targeted chemical agent comprises a DNA carrier, a small molecule chemical agent, and a nucleic acid aptamer EGFR linked to the DNA carrier and biotin, then 2 to 4 biotin molecules are linked to each DNA carrier, and each biotin molecule is linked to either the 5' or 3' end of sequence A or sequence C, respectively; or, B) The target chemical agent comprises a DNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, nucleic acid aptamer EGFR, DNA-based nucleic acid aptamer A15, and optionally biotin, wherein the small molecule chemical is preferably epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a first sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence A in a molar ratio of 1:1 with respect to the DNA carrier; preferably the nucleic acid aptamer AS1411 is linked to the 5' end of sequence A by a base linkage bridge TTTTT; and the nucleic acid aptamer EGFR is linked to the 5' end of sequence B in a molar ratio of 1:1 with respect to the DNA carrier. 'Ends are linked; preferably, the nucleic acid aptamer EGFR is linked to the 5' end of sequence B by a base linkage bridge TTTTT; the nucleic acid aptamer A15 in DNA form is linked to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer A15 is linked to the 5' end of sequence C by a base linkage bridge TTTTT; if the target chemist comprises a DNA carrier, a small molecule chemist and the nucleic acid aptamer AS1411, nucleic acid aptamer EGFR, nucleic acid aptamer A15 in DNA form linked to the DNA carrier, and biotin, then two biotins are linked to each DNA carrier, and each biotin is linked to the 3' ends of sequences A and C respectively; or, C) The target chemical agent comprises a DNA carrier, a small molecule chemical, a DNA-based nucleic acid aptamer AS1411 linked to the DNA carrier, and biotin, preferably the small molecule chemical is epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a first sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer AS1411 is linked to the 5' end of sequence B by a base linkage bridge TTTTT; each DNA carrier has 2 to 4 biotins linked, and each biotin is linked to either the 5' or 3' end of sequence A or sequence C, respectively; or, D) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and nucleic acid aptamer A1 linked to the DNA carrier and nucleic acid aptamer A15 in DNA form, preferably the small molecule chemical agent being epirubicin or its free base or hydrochloride; preferably the DNA carrier comprising a first sequence, with nucleic acid aptamer A1 linked to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably nucleic acid aptamer A1 linked to the 5' end of sequence B by a base linkage bridge TTTTT; preferably nucleic acid aptamer A15 in DNA form linked to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably nucleic acid aptamer A15 linked to the 5' end of sequence B by a base linkage bridge TTTTT; or, E) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and a DNA-based nucleic acid aptamer AS1411, nucleic acid aptamer EGFR, DNA-based nucleic acid aptamer A15, optionally biotin, and oligonucleotide A-miR-21, wherein the small molecule chemical agent is preferably epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a first sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer AS1411 is linked to the 5' end of sequence A by a base linkage bridge TTTTT; and the nucleic acid aptamer EGFR is linked to the DNA carrier The nucleic acid aptamer EGFR is ligated to the 5' end of sequence B in a molar ratio of 1:1 with respect to A; preferably, the nucleic acid aptamer EGFR is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; the nucleic acid aptamer A15 in DNA form is ligated to the 5' end of sequence C in a molar ratio of 1:1 with respect to the DNA carrier; preferably, the nucleic acid aptamer A15 is ligated to the 5' end of sequence C by a base-linking bridge TTTTT; oligonucleotide A-miR-21 is ligated to the 3' end of sequence C in a molar ratio of 1:1 with respect to the DNA carrier; optionally, two biotin molecules are ligated to each DNA carrier, with each biotin molecule ligated to the 3' end of sequence A and the distal end of oligonucleotide A-miR-21 from sequence C, respectively; or, F) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and a DNA-based nucleic acid aptamer TfRA4, a DNA-based nucleic acid aptamer AS1411, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, preferably the small molecule chemical agent being epirubicin or its free base or hydrochloride; preferably the DNA carrier comprising a first sequence, with the DNA-based nucleic acid aptamer TfRA4 linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer TfR A4 is ligated to the 5' end of sequence A by a base-linking bridge TTTTT; the DNA-form nucleic acid aptamer AS1411 is ligated to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer AS1411 is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; the DNA-form nucleic acid aptamer A15 is ligated to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer A15 is ligated to the 5' end of sequence C by a base-linking bridge TTTTT; or, G) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and a DNA-based nucleic acid aptamer TfRA4, a DNA-based nucleic acid aptamer AS1411, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, wherein the small molecule chemical agent is epirubicin or its free base or hydrochloride; the DNA carrier comprises an eleven-set sequence, and the DNA-based nucleic acid aptamer TfRA4 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer TfRA4 is linked to the 5' end of sequence A by a transition base segment SEQ ID NO. 276: 5'-TTGCGGCGAGCGGCGA-3', and the target chemical agent further comprises a complementary sequence SEQ ID NO. 277: 5'-TCGCCGCTCGCCGCTT-3', which is a transition base segment SEQ ID NO. 276: Binds complementary to 5'-TTGCGGCGAGCGGCGA-3', with its 3' end linked to the 5' region of nucleic acid aptamer TfRA4; nucleic acid aptamer AS1411 in DNA form is linked to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably, nucleic acid aptamer AS1411 is linked to the 5' end of sequence B by a base linkage bridge TTTTT; nucleic acid aptamer A15 in DNA form is linked to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably, nucleic acid aptamer A15 is linked to the 5' end of sequence C by a base linkage bridge TTTTT; or, H) The target chemical agent comprises a DNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, a DNA-based nucleic acid aptamer TfRA3, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, preferably the small molecule chemical is epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a first sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer AS 1411 is ligated to the 5' end of sequence A by a base-linking bridge TTTTT; the nucleic acid aptamer TfRA3 in DNA form is ligated to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer TfRA3 is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; the nucleic acid aptamer A15 in DNA form is ligated to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer A15 is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; or, I) The target chemical agent comprises a DNA carrier, a small molecule chemical, and an immunostimulant CPG2006, a nucleic acid aptamer CD40, and a nucleic acid aptamer PD-L1 in DNA form, wherein the small molecule chemical is preferably epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a 9th sequence, and the immunostimulant CPG2006 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the immunostimulant CPG2006 is linked to the base The nucleic acid aptamer CD40 is ligated to the 5' end of sequence A by a ligation bridge TTTTT; the nucleic acid aptamer CD40 is ligated to the 5' end of sequence B in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer CD40 is ligated to the 5' end of sequence B by a ligation bridge TTTTT; the nucleic acid aptamer PD-L1 in DNA form is ligated to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; preferably, the nucleic acid aptamer PD-L1 is ligated to the 5' end of sequence C by a ligation bridge TTTTTT; or, J) The targeted chemical agent comprises a DNA carrier, a small molecule chemical, and an immunostimulant CPG2006, a nucleic acid aptamer CD40, a nucleic acid aptamer PD-L1 in DNA form, and a nucleic acid aptamer C12 linked to the DNA carrier; where the DNA carrier further comprises a complementary sequence D (d sequence) and a complementary sequence E (e sequence), where sequence D is SEQ ID NO. 278: 5'-GCCACCGTGCTACA-3' and sequence E is SEQ ID NO. 279: 5'-CAGCAGCAGCAGCA-3'; the transient base segment SEQ ID NO. 280: 5'-GTAGCACGGTGGC-3' is linked to the 3' end of sequence B in the DNA carrier, and sequence D is linked to the transient base segment SEQ ID NO. 280: Binds complementaryly to 5'-GTAGCACGGTGGC-3', and sequence E binds complementaryly to sequence C of the DNA carrier in a strand-complementary manner; preferably the small molecule chemical is epirubicin or its free base or hydrochloride; preferably the DNA carrier contains the tenth sequence, and the immunostimulant CPG2006 is ligated to the 5' end of sequence A and the 5' end of sequence C, respectively, in a molar ratio of 2:1 to the DNA carrier; preferably the immunostimulant CPG2006 is ligated to the 5' end of sequence A by a base-linking bridge TTTTTT and to the 5' end of sequence C by a base-linking bridge TTTTT; nucleic acid aptamer CD4 0 is ligated to the 5' end of sequence B in a molar ratio of 1:1 with respect to the DNA carrier; preferably, nucleic acid aptamer CD40 is ligated to the 5' end of sequence B by a base-binding bridge TTTTT; nucleic acid aptamer PD-L1 in DNA form is ligated to the 5' end of sequence D in a molar ratio of 1:1 with respect to the DNA carrier; preferably, nucleic acid aptamer PD-L1 is ligated to the 5' end of sequence D by a base-binding bridge TTTTTT; nucleic acid aptamer C12 is ligated to the 5' end of sequence E in a molar ratio of 1:1 with respect to the DNA carrier; preferably, nucleic acid aptamer C12 is ligated to the 5' end of sequence E by a base-binding bridge TTTTT; or, K) The targeted chemical agent comprises a DNA carrier, a small molecule chemical, and an immunostimulant CPG2006, nucleic acid aptamer C12, siRNA oligonucleotide CD47, nucleic acid aptamer PD-L1 in DNA form, and siRNA oligonucleotide PD-L1 linked to the DNA carrier, preferably the small molecule chemical is epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a 9th sequence, siRNA oligonucleotide PD-L1 is linked to the DNA carrier in a molar ratio of 1:1, its sense strand is linked to the 5' end of sequence C in the DNA carrier as a transient base segment, its antisense strand is linked complementaryly to the sense strand as a complementary sequence, and the nucleic acid aptamer PD-L1 in DNA form is linked to the 5' end of the antisense strand of the siRNA oligonucleotide PD-L1; preferably the nucleic acid aptamer PD-L1 is base The siRNA oligonucleotide CD47 is ligated to the 5' end of the antisense strand of siRNA oligonucleotide CD47 by a ligation bridge TTTTT; the immunostimulant CPG2006 is ligated to the 5' end of sequence A in a molar ratio of 1:1 with respect to the DNA carrier; preferably, the immunostimulant CPG2006 is ligated to the 5' end of sequence A by a ligation bridge TTTTT; the siRNA oligonucleotide CD47 is ligated to the 5' end of sequence B as a transient base segment in a molar ratio of 1:1 with respect to the DNA carrier, with its sense strand ligated to the 5' end of sequence B and its antisense strand complementaryly bound to the sense strand as a complementary sequence; the nucleic acid aptamer C12 is ligated to the 5' end of the antisense strand of siRNA oligonucleotide CD47 in a molar ratio of 1:1 with respect to the DNA carrier; preferably, the nucleic acid aptamer C12 is ligated to the 5' end of the antisense strand of siRNA oligonucleotide CD47 by a ligation bridge TTTTT; or, L) The target chemical agent comprises a DNA carrier, a small molecule chemical, and miRNA oligonucleotide A-miR-21, siRNA oligonucleotide TGF-β1, nucleic acid aptamer TfRA4, and nucleic acid aptamer PD-L1 linked to the DNA carrier, preferably the small molecule chemical is epirubicin or its free base or hydrochloride; preferably the DNA carrier comprises a second set sequence, the siRNA oligonucleotide TGF-β1 is in a molar ratio of 1:1 to the DNA carrier, its antisense strand is linked to the 3' end of sequence C as a transient base segment, and the sense strand is linked to the antisense strand as a complementary sequence. Complementarily binding; nucleic acid aptamer TfRA4 is ligated to the 5' end of sequence A in a molar ratio of 1:1 with respect to the DNA carrier; preferably, nucleic acid aptamer TfRA4 is ligated to the 5' end of sequence A by a base-linking bridge TTTTT; nucleic acid aptamer PD-L1 is ligated to the 5' end of sequence B in a molar ratio of 1:1 with respect to the DNA carrier; preferably, nucleic acid aptamer PD-L1 is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; miRNA oligonucleotide A-miR-21 is sequentially ligated to the 5' end of the sense strand of siRNA oligonucleotide TGF-β1 in a molar ratio of 2:1 with respect to the DNA carrier; or, M) The target chemical agent comprises a DNA carrier, a small molecule chemical agent, and nucleic acid aptamer IL-4Ra, siRNA oligonucleotide CD47, nucleic acid aptamer TfRA4 in DNA form, siRNA oligonucleotide PD-L1, and nucleic acid aptamer GPC-1 linked to the DNA carrier, preferably the small molecule chemical agent being epirubicin or its free base or hydrochloride; the DNA carrier comprising a third sequence, with nucleic acid aptamer TfRA4 linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably, nucleic acid aptamer TfRA4 linked to the 5' end of sequence A by a base linkage bridge TTTTT; nucleic acid aptamer GPC-1 linked to the 5' end of sequence C in a molar ratio of 1:1 to the DNA carrier; and siRNA oligonucleotide PD -L1 is linked to the DNA carrier in a molar ratio of 1:1, with its sense strand ligated to the 5' end of sequence B as a transient base segment, and its antisense strand complementaryly bound to the sense strand as a complementary sequence and linked to the 3' end of sequence A; siRNA oligonucleotide CD47 is linked to the DNA carrier in a molar ratio of 1:1, with its sense strand ligated to the 3' end of sequence C as a transient base segment, and its antisense strand complementaryly bound to the sense strand as a complementary sequence; nucleic acid aptamer IL-4Ra is linked to the DNA carrier in a molar ratio of 1:1, with its antisense strand of siRNA oligonucleotide CD47 linked to the 5' end; preferably, nucleic acid aptamer IL-4Ra is linked to the 5' end of the antisense strand of siRNA oligonucleotide CD47 by a base linkage bridge AAAA; or, N) The target chemical agent comprises a DNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, a DNA-based nucleic acid aptamer EGFR, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, preferably the small molecule chemical is gemcitabine; preferably the DNA carrier comprises a fourth sequence, the DNA-based nucleic acid aptamer AS1411 is linked to the DNA carrier in a molar ratio of 1:1 at the 5' end of sequence A, and the DNA-based nucleic acid aptamer EGF R is ligated to the 5' end of sequence B in a molar ratio of 1:1, and the DNA-based nucleic acid aptamer A15 is ligated to the 5' end of sequence C in a molar ratio of 1:1; preferably, gemcitabine is ligated in the form of at least one base insertion to one or more of the following positions: between the DNA-based nucleic acid aptamer AS1411 and sequence A, at the 3' end of sequence A; between the DNA-based nucleic acid aptamer EGFR and sequence B, at the 3' end of sequence B; between the DNA-based nucleic acid aptamer A15 and sequence C, at the 3' end of sequence C; or, O) The target chemical agent comprises an RNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, a DNA-based nucleic acid aptamer EGFR, and a DNA-based nucleic acid aptamer A15 linked to the RNA carrier, preferably the small molecule chemical is 5-fluorouracil; preferably the RNA carrier comprises a fifth sequence, the DNA-based nucleic acid aptamer AS1411 is linked to the RNA carrier in a molar ratio of 1:1 to the 5' end of sequence A, and the DNA-based nucleic acid aptamer EGF R is ligated to the 5' end of sequence B in a molar ratio of 1:1, and the DNA-based nucleic acid aptamer A15 is ligated to the 5' end of sequence C in a molar ratio of 1:1; preferably, 5-fluorouracil is ligated in the form of at least one base insertion to one or more of the following positions: between the DNA-based nucleic acid aptamer AS1411 and sequence A, at the 3' end of sequence A; between the DNA-based nucleic acid aptamer EGFR and sequence B, at the 3' end of sequence B; between the DNA-based nucleic acid aptamer A15 and sequence C, at the 3' end of sequence C; or, P) The targeted chemical agent comprises a DNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, a DNA-based nucleic acid aptamer EGFR, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, preferably the small molecule chemical is paclitaxel; preferably the DNA carrier comprises a sixth sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer Tamer AS1411 is ligated to the 5' end of sequence A by a base-linking bridge TTTTT; the DNA-form nucleic acid aptamer EGFR is ligated to the 5' end of sequence B in a molar ratio of 1:1; preferably, the nucleic acid aptamer EGFR is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; the DNA-form nucleic acid aptamer A15 is ligated to the 5' end of sequence C in a molar ratio of 1:1; preferably, the nucleic acid aptamer A15 is ligated to the 5' end of sequence C by a base-linking bridge TTTTT; or, Q) The targeted chemical agent comprises a DNA carrier, a small molecule chemical, and a DNA-based nucleic acid aptamer AS1411, a DNA-based nucleic acid aptamer EGFR, and a DNA-based nucleic acid aptamer A15 linked to the DNA carrier, preferably the small molecule chemical is maytansine; preferably the DNA carrier comprises a seventh sequence, and the DNA-based nucleic acid aptamer AS1411 is linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier; preferably the nucleic acid aptamer The aptamer AS1411 is ligated to the 5' end of sequence A by a base-linking bridge TTTTT; the nucleic acid aptamer EGFR in DNA form is ligated to the 5' end of sequence B in a molar ratio of 1:1; preferably, the nucleic acid aptamer EGFR is ligated to the 5' end of sequence B by a base-linking bridge TTTTT; the nucleic acid aptamer A15 in DNA form is ligated to the 5' end of sequence C in a molar ratio of 1:1; preferably, the nucleic acid aptamer A15 is ligated to the 5' end of sequence C by a base-linking bridge TTTTT; or, R) The target chemical agent comprises a DNA carrier, a small molecule chemical, and nucleic acid aptamers MUC1, AS1411, and GPC-1 linked to the DNA carrier, preferably the small molecule chemical is gemcitabine; preferably the DNA carrier comprises a ninth sequence, with nucleic acid aptamer MUC1 linked to the 5' end of sequence A in a molar ratio of 1:1 to the DNA carrier, nucleic acid aptamer AS1411 linked to the 5' end of sequence B in a molar ratio of 1:1, and nucleic acid aptamer GPC-1 linked to the 5' end of sequence C in a molar ratio of 1:

1. A targeted chemical agent characterized by the following features.

9. A pharmaceutical composition comprising a target chemical agent according to any one of claims 1 to 8 and a pharmaceutically acceptable carrier and / or excipient.

10. Use of the targeted chemical agent according to any one of claims 1 to 8 in the manufacture of a pharmaceutical product for the treatment of tumors.

11. The use according to claim 10, characterized in that the tumor is an andrological tumor, a gynecological tumor, a respiratory tumor, a digestive tumor, a hematological tumor, a urinary tract tumor, a bone tumor, a nervous system tumor, a dermatological tumor, a general surgical tumor, or a tumor of the five organs (ear, nose, mouth, or ophthalmology); preferably, the andrological tumor is prostate cancer, penile cancer, testicular tumor, or male urethral cancer; the gynecological tumor is ovarian cancer, cervical cancer, endometrial cancer, uterine fibroid, vulvar cancer, or malignant grapeform mole; the respiratory tumor is lung cancer, non-small cell lung cancer, small cell lung cancer, nasopharyngeal cancer, tracheal tumor, lung cancer metastasis, inflammatory pseudotumor, or radiation-induced lung cancer; and the digestive tumor is Systemic tumors include liver cancer, gastric cancer, colorectal cancer, gallbladder cancer, esophageal cancer, rectal cancer, pancreatic cancer, or colon cancer; hematological tumors include leukemia, lymphoma, lymphosarcoma, or multiple myeloma; urinary tract tumors include kidney cancer, bladder cancer, or urinary tract cancer; bone tumors include giant cell tumor, osteochondroma, or osteosarcoma; nervous system tumors include brain tumors, meningiomas, cerebral tuberculoma, pituitary tumors, neuroblastoma, glioblastoma, or glioma; dermatological tumors include skin cancer or malignant melanoma; general surgical tumors include breast cancer, lipoma, thyroid cancer, or thyroid tumor; and ossicular tumors include oral cancer, tongue cancer, laryngeal cancer, middle ear cancer, gingival cancer, or orbital tumors. Preferably, the route of administration in the above use is intratumoral administration, intravenous administration, or intraperitoneal administration; Preferably, the daily dose of this drug in the above-mentioned use is 0.1 μg / kg to 100 mg / kg. A characteristic use.

12. A method for producing a targeted chemical agent according to any one of claims 1 to 8, The process includes forming a targeted nucleic acid carrier comprising an arbitrarily selected oligonucleotide effector molecule and / or an arbitrarily selected immunostimulant by a self-assembly method of sequence; and conjugating a small molecule chemical drug to the targeted nucleic acid carrier to obtain the targeted chemical drug. A manufacturing method characterized by the following features.

13. A manufacturing method according to claim 12, comprising the step of forming the standardized nucleic acid carrier comprising an optionally selected oligonucleotide effector molecule and / or an optionally selected immunostimulant by a sequence self-assembly method, The method involves dissolving at least three sequences having an arbitrarily selected oligonucleotide effector molecule, an arbitrarily selected immunostimulant, and a target molecule in an assembly solution and subjecting them to a denaturation reaction to form a self-assembled crude product; and sequentially purifying, eluting, and evaporating the self-assembled crude product to obtain the targeted nucleic acid carrier comprising an arbitrarily selected oligonucleotide effector molecule and / or an arbitrarily selected immunostimulant; Preferably, the at least three sequences include at least the following three sequences: Sequence A: Sequence A in which an optional oligonucleotide effector molecule, an optional immunostimulant, and an optional target molecule are linked at its ends, either directly or via a base-linking bridge; Sequence B: Sequence B in which an optional oligonucleotide effector molecule, an optional immunostimulant, and an optional target molecule are linked at the end, either directly or via a base-linking bridge; Sequence C: Sequence C in which an optional oligonucleotide effector molecule, an optional immunostimulant, and an optional target molecule are linked at its end, either directly or via a nucleotide linkage bridge; The molar ratio of sequences A, B, and C is 0.90 to 1.10:0.90 to 1.10:0.90 to 1.10, more preferably 1:1:1; if at least three sequences include sequences other than A, B, and C, the molar ratio of each sequence to sequence A is 0.90 to 1.10:0.90 to 1.10, more preferably 1:1; Preferably, if the at least three sequences include sequences other than sequences A, B, and C, these are referred to as complementary sequences, in which case a transition base segment is independently linked to the 5' or 3' end of any of sequences A, B, or C, and the complementary sequences exhibit a base complementary structure with the transition base segment, and the complementary sequences are linked complementaryly to the transition base segment during the self-assembly process; more preferably, the complementary sequences themselves are the sense or antisense strand of an siRNA oligonucleotide effector molecule or a miRNA oligonucleotide effector molecule, and / or the complementary sequences provide a support site for a small molecule target head, target aptamer, or immunostimulant; More preferably, if the at least three sequences include the complementary sequences, the complementary sequences carrying an optional low-molecular-weight target head, an optional target aptamer, and / or an optional immunostimulant are also added to the assembly solution to carry out the denaturation reaction; Preferably, the assembly solution is an aqueous solution of TMS, an aqueous solution of sodium chloride, an aqueous solution of magnesium chloride, or purified water; Preferably, the temperature of the denaturation reaction is 80 to 99°C, more preferably 85 to 99°C, and even more preferably 90 to 99°C; Preferably, after the denaturation reaction is complete, the reaction system is cooled to a holding temperature and a heat retention reaction is performed, and finally the crude product is obtained by cooling; where the holding temperature is 70 to 50°C, more preferably 65 to 55°C, and even more preferably 63 to 57°C; the heat retention reaction time is 3 to 15 minutes, more preferably 3 to 10 minutes, and even more preferably 3 to 5 minutes; Preferably, the cooling rate in the process of lowering the reaction system to the holding temperature is 2 to 10°C / min, more preferably 2 to 6°C / min, and even more preferably 2 to 3°C / min; Preferably, the end temperature of the cooling is 0 to 25°C, more preferably 0 to 15°C, and even more preferably 0 to 4°C; Preferably, the pH of the denaturation reaction is 5.4 to 8.

8. A manufacturing method characterized by the following features.

14. A manufacturing method according to claim 13, If the small molecule chemical is an anthracycline-based chemotherapeutic agent or an acridine-based chemotherapeutic agent, the step of loading the small molecule chemical onto the targeted nucleic acid carrier includes dissolving the targeted nucleic acid carrier comprising the small molecule chemical, a coupling agent, and an optionally selected oligonucleotide effector molecule and / or an optionally selected immunostimulant in a first solvent, reacting under light-shielding conditions to insert the small molecule chemical between adjacent GC base pairs of the targeted nucleic acid carrier in a specific intercalation manner to obtain a crude reaction product, and then sequentially crystallizing, washing, and evaporating the crude reaction product to obtain the targeted chemical; Preferably, the coupling agent is formaldehyde and / or paraformaldehyde; Preferably, the reaction temperature is 0 to 37°C, more preferably 4 to 20°C, and even more preferably 4 to 8°C; Preferably, the pH of the reaction is 6 to 8; Preferably, the reaction time is 24 to 96 hours, more preferably 48 to 72 hours, and even more preferably 72 hours; Preferably, the first solvent is purified water and / or PBS buffer; Preferably, the cleaning agent used in the cleaning step is a halogenated alkane solvent, an ether solvent, or an aromatic solvent, more preferably chloroform, dichloromethane, ether, toluene, or xylene; Preferably, the solvent used in the crystallization step is a lower alcohol-based solvent, more preferably methanol, ethanol, or isopropanol; Preferably, the average number of small molecule chemicals loaded on each of the targeted nucleic acid carriers comprising an optional oligonucleotide effector molecule and / or an optional immunostimulant is 1 to 50, more preferably 10 to 40, and even more preferably 20 to 35; When the small molecule chemical is a pyrimidine-based chemotherapeutic agent, the small molecule target head, oligonucleotide effector molecule, and immunostimulant are linked to the end bases of each sequence of the nucleic acid carrier via a base linkage bridge containing 1 to 10 bases, and the step of binding the small molecule chemical to a targeted nucleic acid carrier comprising an optionally selected oligonucleotide effector molecule and / or an optionally selected immunostimulant includes, in the step of forming the targeted nucleic acid carrier by a sequence self-assembly mechanism, linking the small molecule chemical to at least one sequence of the nucleic acid carrier in the form of substituting at least some of the bases in the base linkage bridge and / or as an extension base and / or as a carrier backbone base substitution, and then completing the binding of the small molecule chemical by a sequence self-assembly mechanism; preferably, the average number of small molecule chemicals carried on each targeted nucleic acid carrier molecule is 1 to 30, more preferably 10 to 20, and even more preferably 12 to 18; When the small molecule chemical is a platinum-based chemotherapeutic agent, a glutamate derivative-based chemotherapeutic agent, a flavonoid-based chemotherapeutic agent, a herbal medicine and its derivatives, a folic acid-based chemotherapeutic agent, or a salicylic acid-based chemotherapeutic agent, the step of conjugating the small molecule chemical to a targeted nucleic acid carrier comprising an optionally selected oligonucleotide effector molecule and / or an optionally selected immunostimulant includes, in the synthesis process of each sequence of the nucleic acid carrier to which the target molecule is linked, introducing a linker into at least one sequence of the nucleic acid carrier to obtain a linker-bound targeted nucleic acid carrier; then dissolving the linker-bound targeted nucleic acid carrier and the small molecule chemical in a second solvent and reacting them; and subsequently cooling, purifying, washing, and evaporating to dryness to obtain the target chemical; Preferably, if the low molecular weight chemical drug itself has a succinimidyl ester group (N-hydroxysuccinimidyl (NHS) ester group), the linker-bound targeted nucleic acid carrier and the low molecular weight chemical drug are dissolved in a second solvent and reacted; if the low molecular weight chemical drug itself does not have an NHS ester group, the process further includes a step of sequentially reacting the low molecular weight chemical drug with bis(2-hydroxyethyl) disulfide and N,N-diisopropylethylamine (DIPEA) before reacting the low molecular weight chemical drug in the second solvent to introduce an NHS ester group into the low molecular weight chemical drug. A manufacturing method characterized by the following features.

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