Delivery device for DNA vaccine

Abstract

The invention relates to an in-vivo delivery device of a DNA vaccine, and belongs to the technical field of vaccine delivery. The guide groove is fixed to one side of the negative pressure tank, the sliding block is slidably mounted in the guide groove, the tail end of the needle is fixed to the sliding block, the bottom end of the infusion tube is connected to the sliding block, the needle is communicated with the infusion tube, the top end of the infusion tube is provided with a connector capable of being in butt joint with an injector quickly, the infusion tube is detachably connected with the injector through the connector, and the top of the negative pressure tank is communicated with an exhaust tube. Traditional Chinese medicine cupping and western medicine intradermal injection means are combined, so that the delivery efficiency of the DNA vaccine is improved, and the immune effect of the DNA vaccine is remarkably improved; tiny negative pressure is provided through the negative pressure tank, so that the skin is shrunk and bulged, a needle is conveniently inserted into skin epithelial tissue at a certain angle, and intradermal injection is completed; according to the vaccination device, the negative pressure tank is arranged in the vaccination device, and then the injector is used for sucking air in the negative pressure tank to provide required negative pressure, so that the operation process is simplified, low-cost disposable equipment without external power is used in the whole process, cross infection of vaccination is avoided, and the vaccination cost is reduced.

Description

Technical Field

[0001] The present invention belongs to the technical field of vaccine delivery, and in particular relates to an in vivo delivery device for DNA vaccines. Background Art

[0002] DNA vaccine is a type of nucleic acid vaccine. It involves directly introducing DNA sequences encoding specific antigens into host cells. The cells then use these DNAs to synthesize antigen proteins, which are recognized by the immune system and trigger an immune response.

[0003] DNA vaccines hold great potential for rapid pandemic response, personalized cancer treatment, and chronic disease prevention and treatment. However, DNA must penetrate the cell membrane to enter cells to be effective. Direct intramuscular injection results in low delivery efficiency and antigenicity. While delivery efficiency can be enhanced through auxiliary equipment and materials such as liposome transfection, gene guns, electroporation, and electric negative pressure devices, these methods are often expensive, limiting their widespread adoption and development.

[0004] Cupping therapy is a traditional Chinese medicine treatment that typically uses glass, bamboo, or plastic jars. By heating or vacuuming, negative pressure is created, which is then applied to the skin. It promotes qi and blood circulation, dispels wind and cold, and regulates yin and yang. It is widely used among the people for its convenience and low cost. From a modern medical perspective, it may also promote blood circulation, relieve muscle tension, activate immune cells, regulate nerve reflexes, and fight inflammation.

[0005] By studying the simple method of using vacuum cupping at the vaccine injection site, detecting and comparing the immune effects of DNA vaccines after injection by different methods, a method to significantly improve the immune effect of DNA vaccines was found. The study showed that increasing negative pressure treatment in situ after intradermal injection of DNA vaccines can increase specific antibody levels and enhance the immune effect of DNA vaccines; however, there is currently a lack of technical devices that combine vaccine injection with vacuum cupping. Cupping can only be performed after injection with a syringe and then through a cupping device. The operation is cumbersome and seriously affects the injection efficiency. The repeated use of electric vacuum negative pressure devices will increase the risk of cross-infection during vaccination. Summary of the Invention

[0006] In order to overcome the problem that the background technology lacks an integrated device that combines intradermal vaccine injection with vacuum cupping, and cupping can only be performed after injection through a syringe and then through a negative pressure device, the operation is cumbersome, which affects the injection efficiency, and the electric negative pressure device is repeatedly used, which increases the risk of cross-infection in vaccination. The present invention provides a DNA vaccine delivery device; it combines Chinese and Western medical methods with the injection device to improve the DNA vaccine delivery efficiency and significantly enhance the immune effect of the DNA vaccine. This invention not only simplifies the operation process, but also uses low-cost disposable equipment without external power throughout the process, which avoids cross-infection in vaccination and reduces the cost of vaccination.

[0007] To achieve the above-mentioned purpose, the present invention is implemented through the following technical solutions: a DNA vaccine delivery device mainly includes a negative pressure tank 1, a guide groove 2, a slider 3, a needle 4, a syringe 5, an infusion tube 6, an exhaust pipe 7, and a one-way valve 8. The guide groove 2 is fixed on one side of the negative pressure tank 1, the slider 3 can be slidably installed in the guide groove 2, the tail end of the needle 4 is fixed on the slider 3, the bottom end of the infusion tube 6 is connected to the slider 3, the needle 4 is connected to the infusion tube 6, and the top of the infusion tube 6 is provided with a joint that can be quickly docked with the syringe 5. The infusion tube 6 is detachably connected to the syringe 5 through the joint, and the DNA vaccine is loaded in the syringe 5; the top of the negative pressure tank 1 is connected to the exhaust pipe 7, the top of the exhaust pipe 7 is provided with a quick-dock joint, the exhaust pipe 7 is provided with a one-way valve 8, and the negative pressure tank 1 is provided with a rubber block 9 opposite to the needle 4 to facilitate the needle 4 to pass through and seal. The negative pressure tank 1 is an elastic tank with good recovery elasticity.

[0008] Furthermore, the infusion tube 6 and the air extraction tube 7 are connected to the syringe 5 through a three-way tube, and a one-way valve 11 is provided on the infusion tube 6.

[0009] Furthermore, the top end of the air extraction tube 7 is connected to a negative pressure syringe 10 via a joint.

[0010] Furthermore, the slider 3 is provided with a protrusion 12 for facilitating pushing and pulling the slider 3 .

[0011] Furthermore, a sealing ring is provided on the edge of the negative pressure tank 1, and an outward-turned flexible edge 13 is provided on the sealing ring.

[0012] Furthermore, the negative pressure tank 1 is a hemispherical structure made of transparent material.

[0013] Furthermore, the rubber block 9 is a rubber sealing plug.

[0014] Beneficial effects of the present invention:

[0015] The present invention achieves the goals of improving DNA vaccine delivery efficiency, enhancing the immune effect of DNA vaccines, reducing the amount of DNA plasmid used, simplifying the DNA vaccination steps, and standardizing the vaccination process. A negative pressure tank provides a slight negative pressure, causing the skin to contract and bulge, making it easier for the injection needle to insert into the skin tissue at a certain angle, achieving standardized intradermal injection. After the intradermal injection is completed, the air in the negative pressure tank is sucked out with a syringe to provide the required negative pressure. After 1-5 minutes of negative pressure treatment, the vaccination is completed. This invention can improve the effectiveness of DNA vaccination while simplifying the operation process and reducing the cost of vaccination. Furthermore, the entire process uses low-cost, non-powered, disposable equipment, avoiding cross-infection during vaccination. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a schematic perspective view of the main axis of embodiment 1 of the present invention.

[0017] Figure 2 It is a perspective view of the second embodiment of the present invention.

[0018] Figure 3 It is a schematic cross-sectional structural diagram of embodiment 2 of the present invention.

[0019] Figure 4 yes Figure 3 Schematic diagram of the locally enlarged structure at point A in the middle.

[0020] Figure 5 It is a schematic diagram of the cross-sectional structure of the infusion state of embodiment 2 of the present invention.

[0021] Figure 6 It is an overhead isometric perspective diagram of embodiment 3 of the present invention.

[0022] Figure 7 This is a schematic diagram of the structure of the pCIS-XBBS plasmid DNA vaccine.

[0023] Figure 8 This is a chart of specific IgG antibody titers in rats after being vaccinated with the new coronavirus DNA vaccine using different immunization methods. DETAILED DESCRIPTION

[0024] In order to make the objectives, technical solutions and beneficial effects of the present invention more clear, preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings to facilitate understanding by technicians.

[0025] Example 1

[0026] The present invention discloses a delivery device for DNA vaccine, which mainly comprises a negative pressure tank 1, a guide groove 2, a slider 3, a needle 4, a syringe 5, an infusion tube 6, an exhaust pipe 7, and a one-way valve 8. The guide groove 2 is fixed on one side of the negative pressure tank 1, the slider 3 can be slidably installed in the guide groove 2, the tail end of the needle 4 is fixed on the slider 3, the bottom end of the infusion tube 6 is connected to the slider 3, the needle 4 is connected to the infusion tube 6, the top of the infusion tube 6 is provided with a joint that can be quickly docked with the syringe 5, the infusion tube 6 is detachably connected to the syringe 5 through the joint, and the DNA vaccine is installed in the syringe 5; the top of the negative pressure tank 1 is connected with an exhaust pipe 7, the top of the exhaust pipe 7 is provided with a joint that can be quickly docked with the syringe 5, the exhaust pipe 7 is provided with a one-way valve 8, and a rubber block 9 is provided on the negative pressure tank 1 opposite to the needle 4 to facilitate the needle 4 to pass through and seal. The pressure cup 1 is an elastic cup with good recovery elasticity. Alcohol is applied to the edge of the negative pressure cup 1. The negative pressure cup 1 is pinched by hand to deform it and then the edge of the negative pressure cup 1 is pressed to the injection position. After the negative pressure cup 1 is released, a small negative pressure is formed in the negative pressure cup 1 due to the recovery deformation of the negative pressure cup 1, causing the skin to shrink and bulge, making it easier for the needle to insert into the skin tissue at a certain angle; the guide groove 2 can achieve smooth sliding of the slider 3 and the needle 4, and insert them into the skin at a smaller angle, and operate the syringe 5 to achieve intradermal injection; after the intradermal injection is completed, operate the slider 3 to withdraw the needle 4, remove the syringe 5 from the infusion tube 6, and dock it on the exhaust tube 7, operate the syringe 5 to suck the air in the negative pressure cup 1 to provide the required negative pressure, thereby achieving the cupping treatment effect, and utilizing the effects of cupping such as promoting blood circulation, relieving muscle tension, and activating immune cells to achieve the purpose of enhancing the immune effect.

[0027] The present invention combines traditional Chinese and Western medical methods to improve the delivery efficiency of DNA vaccines, and the immune effect of DNA vaccines is significantly enhanced; a slight negative pressure is provided by a negative pressure tank to cause the skin to contract and bulge, making it easier for the needle to insert into the skin tissue at a certain angle to achieve intradermal injection; and a syringe is used to draw air from the negative pressure tank to provide the required negative pressure, which not only simplifies the operation process and reduces the cost of vaccination, but also uses low-cost, non-powered, disposable equipment throughout the process to avoid cross-infection during vaccination.

[0028] Working process:

[0029] Apply alcohol on the edge of the negative pressure cup 1, pinch the negative pressure cup 1 by hand to deform it, and then press the edge of the negative pressure cup 1 to the injection position. After releasing the negative pressure cup 1, a small negative pressure is formed in the negative pressure cup 1 under the recovery deformation of the negative pressure cup 1, causing the skin to shrink and bulge, making it easier for the needle to insert into the skin tissue at a certain angle; the guide groove 2 can achieve smooth sliding of the slider 3 and the needle 4, and insert them into the skin at a smaller angle, and operate the syringe 5 to achieve intradermal injection; after completing the intradermal injection, operate the slider 3 to withdraw the needle 4, remove the syringe 5 from the infusion tube 6, and dock it on the exhaust tube 7, operate the syringe 5 to suck the air in the negative pressure cup 1 to provide the required negative pressure, thereby achieving the cupping treatment effect, and utilizing the effects of cupping such as promoting blood circulation, relieving muscle tension, and activating immune cells to achieve the purpose of enhancing the immune effect.

[0030] Example 2

[0031] In this embodiment, based on the first embodiment, the infusion tube 6 and the air extraction tube 7 are connected to the syringe 5 through a three-way tube. A one-way valve 11 is provided on the infusion tube 6. The one-way valve 11 and the one-way valve 8 ensure that the vaccine can only be injected through the infusion tube 6 during injection, and the vaccine will not enter the air extraction tube 7. After the injection is completed, the slider 3 is operated to withdraw the needle 4. When the air is sucked out, due to the action of the one-way valve 11, the syringe 5 can only suck the air in the negative pressure tank 1 to provide the required negative pressure and achieve the cupping treatment effect. Through the connection of the three-way tube and the action of the one-way valve 11 and the one-way valve 8, there is no need to operate, disassemble, or connect the syringe 5.

[0032] In this embodiment, the slider 3 is provided with a protrusion 12 for facilitating pushing and pulling the slider 3 .

[0033] In this embodiment, a sealing ring is provided on the edge of the negative pressure cup 1, and an outward-turned flexible edge 13 is provided on the sealing ring. The outward-turned flexible edge 13 can effectively improve the sealing performance and ensure the cupping effect.

[0034] In this embodiment, the negative pressure cup 1 is a hemispherical structure made of a transparent material, which makes it easy to observe the degree of skin contraction and bulging during the acupuncture operation.

[0035] In this embodiment, the rubber block 9 is a rubber sealing plug, which can meet the requirements of the needle 4 being inserted and forming a seal.

[0036] Working process:

[0037] Apply alcohol on the edge of the negative pressure tank 1, pinch the negative pressure tank 1 by hand to deform it, and then press the edge of the negative pressure tank 1 to the injection position. After releasing the negative pressure tank 1, a small negative pressure is formed in the negative pressure tank 1 under the recovery deformation of the negative pressure tank 1, which makes the skin shrink and bulge, making it easier for the needle to insert into the skin tissue at a certain angle; the slider 3 and the needle 4 can slide smoothly through the guide groove 2 and be inserted into the skin at a smaller angle, and the syringe 5 can be operated to achieve intradermal injection; after the intradermal injection is completed, the slider 3 is operated to withdraw the needle 4; the vaccine can only pass through the one-way valve 11 and the one-way valve 8 during injection. When the infusion tube 6 is injected, the vaccine will not enter the exhaust tube 7. After the injection is completed, the slider 3 is operated to withdraw the needle 4. When aspirating air, due to the action of the one-way valve 11, the syringe 5 can only aspirate the air in the negative pressure cup 1 to provide the required negative pressure and achieve the cupping treatment effect. Through the connection of the three-way pipe and the action of the one-way valve 11 and the one-way valve 8, there is no need to operate or disassemble the syringe 5 to provide the required negative pressure and achieve the cupping treatment effect. The effects of cupping such as promoting blood circulation, relieving muscle tension, and activating immune cells are utilized to achieve the purpose of enhancing the immune effect.

[0038] Example 3

[0039] In this embodiment, based on the first and second embodiments, the top ends of the infusion tube 6 and the exhaust tube 7 are respectively connected to the syringe 5 and the negative pressure syringe 10 through joints. The vaccine is injected through the syringe 5, and the negative pressure is sucked out through the negative pressure syringe 10. The vaccine injection and the negative pressure suction are each carried out with one syringe, and they do not affect each other in practice. The negative pressure syringe 10 is used to facilitate the adjustment of the negative pressure in the negative pressure tank 1 before the needle is inserted, and then the height of the skin contraction and bulging of the negative pressure tank 1 is adjusted to ensure the effect of intradermal injection and improve the injection efficiency.

[0040] Example 4

[0041] The following animal studies were used to demonstrate the beneficial effects of negative pressure aspiration of the injection site after vaccination.

[0042] 1. Materials and Methods

[0043] 1. Preparation of Recombinant DNA Plasmid

[0044] The DNA vaccine is a pCIS-XBBS recombinant plasmid. Based on the amino acid sequence of the spike protein S of the new coronavirus XBB.1 mutant strain, the optimized nucleotides and their encoded amino acid sequences are shown in Table 1 below. The HindIII and BamHI restriction sites were used to connect the pCISMA animal DNA vaccine expression vector ( Figure 7 ). Transform DH5α Escherichia coli host cells for amplification, and extract the plasmid using an endotoxin-free plasmid extraction kit for later use.

[0045] Table 1 Amino acid sequence and optimized nucleotide sequence of spike protein S of the novel coronavirus XBB.1 mutant strain

[0046]

[0047]

[0048] 2. Experimental Animals and Immunity Tests

[0049] 2.1 Experimental animals were 8-10 week old Sprague-Dawley rats.

[0050] 2.2 After the rats were depilated at the injection site (thigh), 400 μg of p CIS-XBBS DNA vaccine was administered intramuscularly with vacuum treatment, intradermally with vacuum treatment (vacuum treatment for 30 seconds), or intradermally without vacuum treatment. Blood was collected from the tail vein two weeks later, and serum was isolated for antibody testing.

[0051] 2.3 ELISA enzyme-linked immunosorbent assay indirect method to detect anti-S protein specific IgG antibody titer

[0052] Each well of a 96-well microtiter plate was coated with 0.2 μg of recombinant novel coronavirus mutant XBB.1S1 protein (coating solution: 15 mM Na₂CO₃ and 35 mM NaHCO₃, pH 9.6) and incubated overnight at 4°C. The coating solution was discarded, and the microtiter plates were washed with PBST (0.5% TWEEN-20 / PBS) and blocked with 2% BSA / PBST at 30°C for 1 hour. Animal serum was serially diluted in PBS and added to the sample wells, incubated at 30°C for 1 hour. After washing once with PBST, a 1:5000 dilution of horseradish peroxidase (HRP)-conjugated goat anti-rat IgG (Thermo Fisher) was added and reacted at 30°C for 1 hour. Color was developed using TMB substrate, the reaction was stopped with sulfuric acid, and the absorbance was measured at 450 nm using a microtiter plate reader. The S protein-specific IgG titer was determined based on absorbance, with 2.1 times the absorbance of blank serum as the cutoff value. The anti-novel coronavirus S protein-specific IgG antibody titer was determined by the endpoint dilution method, and the maximum serum dilution with an absorbance greater than the cutoff value was taken as the titer level of anti-S-specific antibodies.

[0053] 3. Statistical Analysis

[0054] The antibody titers between the groups were compared using the t-test, and a P value less than 0.05 was considered to be significantly different.

[0055] 2. Experimental Results

[0056] The experimental results are as follows Figure 8 As shown in the figure ("-" represents direct syringe injection without vacuum treatment, and "+" represents negative pressure cupping treatment), intradermal injection of DNA vaccine is better than intramuscular injection. The superimposed negative pressure treatment of the device invention can significantly improve the immune effect after DNA vaccination (P<0.001).

[0057] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A DNA vaccine delivery device, characterized in that: The DNA vaccine delivery device comprises a negative pressure tank (1), a guide groove (2), a slider (3), a needle (4), a syringe (5), an infusion tube (6), an air extraction tube (7), and a one-way valve (8). The guide groove (2) is fixed on one side of the negative pressure tank (1), the slider (3) is slidably installed in the guide groove (2), the tail end of the needle (4) is fixed on the slider (3), the bottom end of the infusion tube (6) is connected to the slider (3), the needle (4) is connected to the infusion tube (6), and the top end of the infusion tube (6) is connected to the syringe (5). A connector is provided that can be quickly connected to the syringe (5), and the infusion tube (6) is detachably connected to the syringe (5) through the connector, and the DNA vaccine is placed in the syringe (5); the top of the negative pressure tank (1) is connected to an exhaust pipe (7), the top of the exhaust pipe (7) is provided with a connector that can be quickly connected, and a one-way valve (8) is provided on the exhaust pipe (7). A rubber block (9) is provided on the negative pressure tank (1) opposite to the needle (4) to facilitate the needle (4) to pass through and seal. The negative pressure tank (1) is an elastic tank with good recovery elasticity.

2. The DNA vaccine delivery device according to claim 1, wherein: The infusion tube (6) and the air extraction tube (7) are connected to the syringe (5) via a three-way tube, and a one-way valve (11) is provided on the infusion tube (6).

3. The DNA vaccine delivery device according to claim 1, wherein: The top end of the air extraction tube (7) is connected to a negative pressure syringe (10) via a joint.

4. A DNA vaccine delivery device according to any one of claims 1 to 3, characterized in that: The slider (3) is provided with a protrusion (12) for facilitating pushing and pulling the slider (3).

5. The DNA vaccine delivery device according to claim 4, characterized in that: The edge of the negative pressure tank (1) is provided with a sealing ring, and the sealing ring is provided with an outward-turned flexible edge (13).

6. A DNA vaccine delivery device according to any one of claims 1, 2, 3, and 5, characterized in that: The negative pressure tank (1) is a hemispherical structure made of a transparent material.

7. The DNA vaccine delivery device according to claim 6, characterized in that: The rubber block (9) is a rubber sealing plug.

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