CN112156190A - Nano drug loading system based on ship-shaped DNA origami - Google Patents
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Abstract
The invention belongs to the field of drug delivery, and discloses a nano drug delivery system based on ship-shaped DNA origami, which comprises a ship-shaped DNA origami main body (101), a capturing chain (102) which is positioned at the groove part of the ship-shaped DNA origami main body and extends out, thrombin (103) which is positioned at the groove part of the ship-shaped DNA origami main body, targeting chains (105) which are positioned at two ends of the ship-shaped DNA origami main body and extend out, and a targeting aptamer (106) which is fixed on the targeting chains and is used for targeting and identifying nucleolin; wherein, the surface of the thrombin is modified with a connecting chain (104), and the connecting chain is used for being combined with the capturing chain; the boat-shaped DNA origami main body is formed by bonding a single M13 loop-shaped single strand through a staple chain. According to the invention, by improving the key shape design and detail structure composition of the DNA origami nano medicine carrying system, the problems of complex metamorphism of a nano structure, error rate in medicine carrying and the like in the prior art can be solved.
Description
Technical Field
The invention belongs to the field of drug delivery, and particularly relates to a nano drug delivery system based on ship-shaped DNA paper folding.
Background
The paper "Folding DNA to create nanoscales flaps and patterns" was published by the American scientist Paul W.K.Rothemund in Nature journal as an independent author in 2006 and was considered to be the beginning of the DNA origami. Rothemund uses a modified M13 phage nucleic acid, circular single-stranded DNA molecule as the backbone chain, selects specific sites of circular DNA, designs 200 short-stranded DNA (called staple chain), and links the specific sites of circular single-stranded DNA molecule together by means of base complementary hybridization, thereby folding the circular DNA molecule into a nanostructure of arbitrary shape. Meanwhile, Rothemund extends the staple chain of the designated site to the outside of the nano structure, and then DNA molecules with secondary structures are arranged on the surface of the paper folding structure through complementary hybridization, thereby proving the space addressing capability of the DNA paper folding structure. As shown in fig. 1.
DNA hybridization can be used to assemble DNA molecules to the surface of the paper folding, complementary to the staple strand extension site, as long as they are capable of attaching DNA molecules to biomolecules. By utilizing the characteristic of the DNA origami, researchers arrange biomolecules capable of inhibiting the growth of tumor cells and aptamers capable of identifying the tumor cells in a targeted mode on the surface of the DNA origami, so that a technology of drug loading and targeted transportation based on the DNA origami structure is formed. A drug loading strategy using a variable DNA origami structure is reported in the article A DNA nanorobot functions as a cancer therapeutic in response to a molecular trigger in vivo published in the Nature Biotechnology journal. This document links thrombin protein and DNA molecules and loads thrombin onto the rectangular origami of the Rothemund invention by complementary pairing. Through the design, the rectangular folded paper is rolled into a cylindrical structure by utilizing DNA hybridization, and thrombin is wrapped in the cylindrical structure, so that the thrombin is ensured not to influence normal blood vessels before being released, namely, the normal parts of an organism are not attacked. It is noteworthy that the DNA molecule triggering the origami curl consists of partially complementary DNA duplexes (Y-configuration DNA), one of which consists of the aptamer AS1411, which is able to specifically recognize nucleolin and bind to it to form a G tetramer, while the duplexes unwind and the rectangular DNA origami unfold. The nucleolin protein is selectively expressed on the surface of the endothelial cells of the tumor blood vessels which are actively proliferated, so that the medicine carrying device reported in the document can specifically identify the tumor position and expose thrombin in the blood vessels related to the tumor, and the thrombin can assist platelets and fibrin in blood to quickly form thrombus to block the blood vessels of the tumor, so that the nutrition and oxygen of the tumor are deprived, and a large amount of tumor cells are killed. In order to enhance targeting of the drug delivery system, researchers also distributed a separate aptamer AS1411 at both ends of the paper fold. As shown in fig. 2.
The method for treating the tumor utilizes the natural biocompatibility and low toxicity of the DNA material to transport the thrombin molecules to the blood vessels related to the tumor in a targeted way, thereby effectively inhibiting the growth of tumor cells. Thrombin-selective occlusion of blood vessels is also considered an attractive approach to tumor therapy. First, vascular occlusion can rapidly induce thrombosis in tumor vessels, which then exert its effect within hours and reduce the risk of drug resistance development. Furthermore, because all solid tumors supply blood vessels with essentially the same blood, vascular occlusion is considered a strategy that can be used for many types of cancer. This method is also the first example of the use of a DNA origami drug delivery system in mammals, and achieves therapeutic results in both mouse and bama minipig intravenous injection experiments.
This technique still has some drawbacks: firstly, the paper folding allosteric mechanism is complex, when the medicine is wrapped by the curled paper folding, the medicine has certain randomness, as shown in fig. 3, thrombin is exposed on the outer surface with certain probability, as shown in fig. 3, and the by-product is difficult to remove, so that the effective yield is reduced, and the exposed thrombin attacks blood vessels of normal parts; second, single ply origami are considered to lack rigidity, limited ability to carry and protect drugs; thirdly, thrombin is not released when reaching the tumor site and still arranged on the surface of the DNA paper folding, and the steric hindrance formed by the paper folding structure may affect the curative effect.
Disclosure of Invention
Aiming at the defects or improvement requirements of the prior art, the invention aims to provide a nano medicine carrying system based on ship-shaped DNA origami, wherein the key shape design and the detailed structure composition of the DNA origami nano medicine carrying system are improved, an M13 annular single chain is used for forming a ship-shaped DNA origami main body through a stitching needle chain, thrombin is fixed at a ship-shaped groove part, and meanwhile, targeting aptamers for identifying nucleolin in a targeting way are arranged at the two ends of the ship-shaped DNA origami main body, so that the technical problems that the nano structure is complex, the medicine carrier is not rigid enough, the protection effect on the medicine is not good, the medicine is not completely released and the like, the curative effect is possibly influenced and the like can be solved.
In order to achieve the above object, according to the present invention, there is provided a nano drug delivery system based on ship-shaped DNA origami, which is characterized by comprising a ship-shaped DNA origami main body (101), a capturing chain (102) extending out of the groove of the ship-shaped DNA origami main body (101), thrombin (103) extending out of the groove of the ship-shaped DNA origami main body (101), targeting chains (105) extending out of the two ends of the ship-shaped DNA origami main body (101), and targeting aptamers (106) fixed on the targeting chains (105) for targeting and recognizing nucleolin; wherein, the surface of the thrombin (103) is modified with a connecting chain (104), the connecting chain (104) is used for being combined with the capturing chain (102), and the capturing chain (102) can also be used for targeted recognition of nucleolin; the boat-shaped DNA origami main body (101) is formed by bonding a single M13 loop-shaped single strand through a staple chain.
As a further preferable aspect of the present invention, the boat-shaped DNA origami main body (101) is formed of 34 double spirals, both ends of which are formed with a cross section formed by stacking the 34 double spirals in a honeycomb manner, and the groove portion is formed by a void of a portion of the 6 double spirals stacked therein; the length of the boat-shaped DNA origami main body (101) is 76nm, and the length of the groove part is 30 nm.
As a further preferred aspect of the present invention, the targeting aptamer (106) for targeting recognition of nucleolin is aptamer AS 1411.
As a further preferred aspect of the present invention, there are 8 targeting chains (105) in the nano drug delivery system of any one ship-shaped DNA origami, wherein 4 targeting chains (105) are respectively disposed at two ends of the ship-shaped DNA origami main body (101), and these 8 targeting chains (105) are used for binding with the targeting aptamer (106).
In a further preferred embodiment of the present invention, 2 of the above-mentioned connecting chains (104) are modified on the surface of any thrombin (103).
As a further preferred aspect of the present invention, the nucleotide sequence of the connecting strand (104) is shown in SEQ ID No. 200.
As a further preferable aspect of the present invention, in any one of the nano drug delivery systems of ship-shaped DNA origami, the number of the thrombin (103) is not more than 3.
In a further preferred embodiment of the present invention, in any one of the nano drug delivery systems of ship-shaped DNA origami, the number of thrombin (103) is 3.
As a further preferred aspect of the present invention, the nano drug delivery system of any ship-shaped DNA origami has 6 capturing chains (102), and each 2 capturing chains (102) are used for matching and combining with one thrombin (103).
Through the technical scheme, compared with the prior art, the boat-shaped DNA origami main body is used, the special boat-shaped design is utilized, thrombin is fixed at the boat-shaped groove part in a matched mode, the targeting aptamers for targeted nucleolin identification are arranged at the two ends of the boat-shaped DNA origami main body, and the original boat-shaped DNA origami structure has a three-dimensional structure, so that the rigidity of a DNA origami carrier can be effectively improved, and the better protection before the targeted release of a medicament is ensured; the assembly sites and the assembly mode are reasonably designed, the groove part of the boat-shaped folded paper is used for protecting the medicine, and the allosteric operations such as folding, curling and the like of the DNA nano structure are not needed, so that the transfer accuracy is ensured; an aptamer recognition release mechanism is designed, nucleolin can be recognized, thrombin can be released, the effect of the thrombin in a specified position is fully exerted, and the aptamer recognition release mechanism has a very large potential application value.
The invention designs and prepares the original ship-shaped nano structure, skillfully utilizes the characteristic of combining the aptamer (such AS AS1411) and the nucleolin selectively expressed by the tumor, and designs the obtained drug delivery controlled release system which can not only identify the tumor cells in a targeted way, but also release thrombin near the blood vessels related to the tumor. The present invention preferably uses a capturing strand and a targeting strand of a specific nucleotide sequence, which are involved in the construction of a DNA origami structure on the one hand, and extend a part of the sequence to the surface of the origami structure, and the Thrombin Thrombin and the aptamer having the DNA sequences are immobilized by complementary pair hybridization. Also, for the targeting strand used for immobilization of the aptamer, the aptamer sequence does not participate in hybridization with the extended targeting strand, but is immobilized by hybridization with the targeting strand via the extended portion.
The invention also can effectively ensure the drug-loading effect by optimally designing the size and the structure of the ship-shaped DNA origami. The boat-shaped folded paper is used as a medicine carrier, and the space size of the groove part is a very important parameter. The larger the groove design, the greater the amount of medication that can be carried by a single folded sheet, but an excessively large groove can adversely affect the stability of the folded sheet. Therefore, considering the bearing capacity of the ship-shaped folded paper and the stability requirement of the folded paper structure, the invention preferably sets the size parameters of the ship-shaped folded paper as follows: the boat-shaped DNA origami main body is composed of 34 double spirals, the two ends of the boat-shaped DNA origami main body are provided with sections formed by the 34 double spirals in a honeycomb stacking mode, the groove parts are formed by partial vacancy of 6 double spirals (namely a group of honeycomb structures) stacked in the 34 double spirals, the length of the boat-shaped DNA origami main body is 76nm, the length of the groove parts is 30nm (in the DNA origami structure, factors such as intermolecular repulsion are considered, each double spiral is 2.25nm in diameter, each thread pitch averagely comprises 10.5 base pairs, namely the boat-shaped DNA origami main body formed by stacking the 3.57 nm.34 double spirals in a honeycomb stacking mode is 76nm long, the section is 18nm multiplied by 12nm, the groove part is 30nm long, and the section is approximate to a regular hexagon with the side length of 4.5 nm.
Drawings
Fig. 1 is a prior art schematic.
Fig. 2 is a prior art schematic.
FIG. 3 is a schematic of a prior art analysis.
FIG. 4 is a schematic diagram of the overall structure of the original ship-shaped DNA origami of the invention after loading thrombin and aptamer AS 1411.
FIG. 5 is a schematic diagram of drug delivery system targeting recognition of nucleolin selectively expressed by tumor cells and release of thrombin.
FIG. 6 is a schematic side view of the double spiral arrangement in the design of the ship-shaped DNA origami of the present invention. In the figure, 34, 35 are reserved positions for showing the extension of the capturing chain and its position.
FIG. 7 is a schematic sequence diagram of the ship-shaped DNA origami design in the present invention. In the figure, the uppermost numbering is that of the DNA double helix and corresponds to that of FIG. 6. Wherein, the No. 4, 5, 6, 7, 16 and 17 double helix is the position of the groove part. The lower main pattern portion, the vertical line segments represent DNAs, and the horizontal line segments represent connections between DNAs. 34. The reserved position number 35 corresponds to the capture chain.
FIG. 8 is a TEM representation of the preparation of boat-shaped DNA origami according to the invention.
The meaning of the reference numerals in fig. 4 is as follows: 101 is a boat-shaped DNA origami main body, 102 is a capturing chain, 103 is thrombin, 104 is a connecting chain, 105 is a targeting chain, and 106 is a targeting aptamer.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.
Generally, the nano drug delivery system based on the ship-shaped DNA origami can be designed and prepared according to the following steps:
firstly, a ship-shaped DNA origami structure is designed and prepared. Reasonably arranging the skeleton chains to form a ship-shaped three-dimensional conformation; can design the staple with specific sitesA chain, fixing the skeleton chain; designing sites extending out of an aptamer (such AS AS1411) at the position of the boat-shaped paper folding groove, hybridizing thrombin with a DNA molecule partially complementary with the thrombin, and releasing the thrombin under the triggering of nucleolin; the two ends of the boat-shaped folded paper are designed with the extension of a staple chain, and a nucleic acid aptamer (such AS AS1411) is connected by a DNA chain hybridization method for improving targeting. The designed skeleton chain sequence and the designed staple sequence can be mixed according to the ratio of 1:10 and are 1 XTAE/Mg2+Mixing the solution, and carrying out PCR temperature-changing reaction to obtain the DNA origami structure. The specific requirements of the PCR temperature-variable reaction can be shown in Table 1.
Table 1: PCR temperature-changing reaction temperature control program
| Temperature[℃] | Duration[min] |
| 85 | 5 |
| 65-61 | 5 |
| 60-51 | 60 |
| 50-38 | 20 |
| 37-25 | 10 |
| 25 | forever |
Second, the thrombin loading regime is designed. By referring to the method in the background art for connecting thrombin and DNA molecules, namely, using a sulfo-SMCC cross-linking agent to connect thrombin and DNA; the DNA molecules connected with the thrombin protein are partially complementary with the aptamer sequences protruding from the groove to form a Y-shaped DNA double strand, as shown in FIG. 4; considering the molecular weight (-37 KDa) and the steric hindrance of the thrombin protein, 3 thrombin proteins are preferably designed in each notch part of the ship-shaped origami; in order to fix the thrombin closer to the inside of the groove so AS to form better protection, and simultaneously fully expose the Y-shaped DNA double chain, so that nucleolin is convenient to combine with the aptamer AS1411 to release the thrombin, the invention adopts a chain hybridization mode shown in figure 4.
By comprehensively considering the molecular weight of the thrombin protein and the morphological characteristics of the DNA origami, 6 strands of double helix outside the cylindrical origami are hollowed out (as shown in figure 4); when designing a thrombin capture site, considering a steric hindrance factor, reserving a gap of about 5nm between the thrombins; when capturing thrombin by strand hybridization, the roots of the thrombin linked to DNA may be aligned with the roots of the capturing strands protruding from the DNA origami to better hide the thrombin in the recess.
Finally, in order to improve targeting of the drug-loaded device, a staple chain (i.e., the targeting chain 105) is designed to extend out of both ends of the boat-shaped DNA origami for fixing the aptamer AS 1411. It is noted that the aptamer sequence does not participate in hybridization with the protruding staple strand, but is immobilized by hybridization with the staple strand via an extended portion, as shown in FIG. 4.
In addition, for each boat-shaped folded paper structure, 3 thrombin molecules can be immobilized at the groove portion, 4 AS1411 can be immobilized at each end of the folded paper, and 8 aptamer AS1411 can be immobilized in total. The two ends of the boat-shaped DNA origami main body are preferably provided with 8 targeting chains, and mainly, the origami structure can better load the 8 targeting chains on one hand, and the arrangement of the 8 targeting chains can also better realize the targeting effect on the other hand.
The following are specific examples:
example 1
Fig. 4 and fig. 5 show schematic diagrams of two stages of thrombin loading and thrombin releasing of the drug delivery system of the invention. As shown in fig. 4, the drug delivery system consists of the following components: the kit comprises a boat-shaped DNA origami main body 101, a skeleton chain extending out of the origami groove part, a capture chain 102, loaded medicine thrombin 103, a connecting chain 104 modified on the thrombin 103 and used for combining the capture chain 102, skeleton chains extending out of two ends of the origami main body, a targeting chain 105 and a targeting aptamer 106 fixed on the targeting chain 105 and used for targeting and recognizing nucleolin.
The structure of the original ship type DNA origami is completed on the basis of the cadano software, and the design interfaces are shown in FIGS. 6 and 7. Firstly, designing the double helix arrangement of the lateral DNA molecules according to the geometric conformation of the paper folding structure. And then arranging the skeleton chains on the development diagram design interface. According to the correspondence between the expanded view and the side view, the groove portion is reserved. Because the entity of the framework chain adopted in the preparation of the DNA origami is the M13 circular single chain, the whole framework chain needs to be designed into a complete closed loop. The arrangement of the backbone strands and the side double helix arrangement determine the shape of the entire DNA origami. The chain of staples is then automatically filled using software and the resulting chain of staples is modified, including modification of the chain of staples with extended extension of the edge portion of the structure (to weaken intermolecular stacking bonds, avoid undesired connections between folds) and too long or too short of a staple chain. The basic shape of the ship-shaped DNA origami is ensured by the design, and the sites can be selected and added with the skeleton chain with special function after the completion.
Considering the size of thrombin and the space of the groove part of the boat-shaped paper folding, it is preferable to determine 3 sites for fixing thrombin for each DNA paper folding design, and 2 extended backbone chains for increasing the probability of capturing thrombin are designed for each site, wherein the extended backbone chains are the capture chains in FIG. 4. In selecting the specific site for extension of the capture strand, the spatial considerations of the DNA double helix need to be taken into account to ensure that the capture strand extends beyond the notch. The skeleton chain is extended out of the proper position of the two ends of the folded paper, and the targeting chain in the figure 4 can be formed.
After the paper folding design is completed, the DNA sequence can be output. The sequence of the backbone chain can be automatically obtained by the cadano software by selecting a marker head and tail on the backbone chain, and filling the M13 sequence at the 5' end. It is noted that thymine T is filled in the common skeleton chain extending out of the edge of the paper folding structure; the projecting parts of the projecting skeleton chain with special function need to be designed separately. Wherein, the capture chain can refer to the design scheme in the background technical literature; targeting strands can be designed by NUPACK software, as shown in tables 2.1 to 2.3.
TABLE 2.1 boat-shaped DNA origami common staple chain
TABLE 22 boat-shaped DNA origami protruding out of the staple chain
TABLE 2.3 other sequences
M13 was purchased from Tilibit nanosystems, Germany, and all remaining DNA sequences were synthesized by Biotechnology, Inc. The synthetic DNA strand requires a lysis treatment. Firstly, centrifuge the test tube containing DNA, set the rotation speed 8000rpm, time 3 min. Deionized water was then added to the tubes in the recommended volumes for the order. Vortex and shake for 3min before using a low speed centrifuge to funnel the solution to the bottom of the tube. Each DNA strand was tested for concentration using a ultramicro spectrophotometer Nano Drop 2000.
After the DNA strand concentration test is completed, the staple strands need to be mixed. Mixing common staple chains according to the concentration of the DNA chains tested in the previous step and the quantity of the substances; special-purpose staple chains (including the capture chain and the targeting chain) were mixed at a ratio of 1.5 times the amount of the substance. Thereby forming a staple chain mixture.
mu.L of M13 strand (2 nM), 10. mu.L of the mixture of staple strands and 15pmol of the targeted aptamer strand (15 nM) were mixed in 1 XTAE solution (2-valent magnesium ion, pH 8.0) to constitute a 100. mu.L solution system and placed in a small-sized EP tube (volume 200. mu.L). Using a PCR instrument, annealing was started from 95 ℃ to room temperature, and the specific procedure used can be as shown in Table 1.
After PCR annealing, the DNA origami needs to be purified. This experiment used an ultracentrifuge tube to separate the target product from the excess staple chain. An ultracentrifuge tube (MWCO, Amicon, Millipore) containing a 100k filter was wetted with 1 xtae buffer, the origami sample to be purified was then added to the tube, the tube was filled to full volume (500uL) with 1 xtae, centrifuged at 12000rpm using a centrifuge, the sample was passed through the filter, and the excess DNA strands were filtered off, leaving the pure DNA origami structure. The centrifugation process can be repeated 5 times.
The structure of the purified DNA origami can be represented by a TEM image. Firstly, taking a common carbon supporting film (a mesoscope instrument) as a sample supporting net, and carrying out plasma cleaning on the sample supporting net: and placing the carbon film on the surface of the glass plate, inserting the glass plate into a vacuum cavity of the plasma cleaning instrument, closing the air hole of the cover, and covering the sampling opening of the vacuum cavity. And opening a vacuumizing switch, and vacuumizing the vacuum cavity for 1.5 min. And opening the plasma cleaning instrument and adjusting to a low gear. After plasma cleaning for 30 seconds, the air inlet hole is opened slowly, so that air enters the vacuum cavity slowly, and the carbon film is prevented from being blown away by violent air flow. After the glass plate was removed, the cleaned copper mesh was transferred to an analytical filter paper for use. Then, holding a nipper pliers with one hand, lightly pressing a carbon film on the filter paper, holding a pipette with one hand, dropping 2 mu L of the sample of the paper folding to be detected on the surface of the copper mesh, slightly fluctuating the solution drop with the pipette head to assist the absorption of the solution drop by the filter paper, and simultaneously keeping the target structure in the carbon film through the carbon film. Finally, if DNA is to be stained, 2. mu.L of uranyl acetate (mesoscope), which is dropped onto the carbon film in the same manner as the dropping method, is taken. And standing the copper net with the dripped sample for 30 minutes to be used for observation of an electron microscope.
Referring to the background technology, the method for linking thrombin and DNA comprises the following steps: linking thrombin 103 and linker 104 using a sulfo-SMCC crosslinker; the concentration of purified DNA-thrombin and the DNA labeling rate were estimated by measuring the absorbance at 260nm and 280 nm. The ratio of DNA to thrombin marker was estimated to be 2.5. + -. 0.8 in the literature. The ligation of DNA-labeled thrombin to the DNA origami structure takes approximately 60-100 min.
FIG. 8 shows the TEM representation result of the boat-shaped DNA origami in this example, and it can be seen from FIG. 8 that the topography of the boat-shaped DNA origami is substantially consistent with the design, the groove portions are clearly visible, and the size is consistent with the design.
The reagents used in the present invention are all commercially available reagents.
It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (8)
1. A nano drug delivery system based on ship-shaped DNA origami is characterized by comprising a ship-shaped DNA origami main body (101), a capturing chain (102) which is positioned at the groove part of the ship-shaped DNA origami main body (101) and extends out, thrombin (103) which is positioned at the groove part of the ship-shaped DNA origami main body (101), targeting chains (105) which are positioned at two ends of the ship-shaped DNA origami main body (101) and extend out, and a targeting aptamer (106) which is fixed on the targeting chains (105) and is used for targeting and recognizing nucleolin; wherein, the surface of the thrombin (103) is modified with a connecting chain (104), the connecting chain (104) is used for being combined with the capturing chain (102), and the capturing chain (102) can also be used for targeted recognition of nucleolin; the boat-shaped DNA origami main body (101) is formed by bonding a single M13 loop-shaped single strand through a staple chain.
2. The nano drug delivery system based on ship-shaped DNA origami as claimed in claim 1, wherein the ship-shaped DNA origami main body (101) is composed of 34 double spirals, both ends of the ship-shaped DNA origami main body form a cross section formed by the 34 double spirals in a honeycomb type stacking manner, and the groove part is formed by the vacancy of 6 double spiral parts stacked in the groove part; the length of the boat-shaped DNA origami main body (101) is 76nm, and the length of the groove part is 30 nm.
3. The ship-shaped DNA origami-based nanoparticulary system of claim 1, wherein the targeting aptamer (106) for targeted recognition of nucleolin is aptamer AS 1411.
4. The nano-drug delivery system based on the ship-shaped DNA origami as claimed in claim 1, wherein the nano-drug delivery system of any one ship-shaped DNA origami has 8 targeting chains (105), wherein 4 targeting chains (105) are respectively arranged at two ends of the ship-shaped DNA origami main body (101), and the 8 targeting chains (105) are used for being combined with the targeting aptamer (106).
5. The nano drug delivery system based on ship-shaped DNA origami as claimed in any one of claims 1 to 4, wherein 2 connecting chains (104) are modified on the surface of any one thrombin (103).
6. The nano drug delivery system based on ship-shaped DNA origami as claimed in any one of claims 1 to 5, wherein the number of thrombin (103) in the nano drug delivery system of any one ship-shaped DNA origami is not more than 3.
7. The nano drug delivery system based on ship-shaped DNA origami as claimed in claim 1, wherein in any one nano drug delivery system of ship-shaped DNA origami, the number of thrombin (103) is 3.
8. The nano-drug delivery system based on ship-shaped DNA origami as claimed in claim 5, wherein the nano-drug delivery system of any ship-shaped DNA origami has 6 capturing chains (102), and each 2 capturing chains (102) are used for matching and combining with one thrombin (103).
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