CN118002219A - Nanometer medicine preparation equipment based on micro-fluidic technology - Google Patents

Abstract

The invention relates to the technical field of microfluidic pharmaceutical equipment, in particular to nano-drug preparation equipment based on a microfluidic technology, which comprises a chip box, a drug input assembly, a power assembly, a collection assembly and a collection assembly support piece, wherein the power assembly comprises a first power device and a second power device, the first power device is in driving connection with the drug input assembly, the second power device is in driving connection with the collection assembly support piece, and the collection assembly is detachably connected with the collection assembly support piece. The invention has high degree of automation and good use experience of users, and the universal chip conversion box used can be adapted to different kinds of chips, thereby improving the application range of products.

Description

Nanometer medicine preparation equipment based on micro-fluidic technology

Technical Field

The invention relates to the field of scientific instruments, in particular to microfluidic equipment, and especially relates to nano-drug preparation equipment based on a microfluidic control technology.

Background

Under the great background of the epidemic of new world crown, the capability of developing, producing and supplying vaccines measures the comprehensive strength of a country. The research and development path of the traditional vaccine has the problems of high cost, long period, insufficient effectiveness and the like, so that some developing countries are prohibitive; the technical route of the mRNA vaccine is widely applied in the advantages of low cost, short period, high safety and effectiveness, and the like, and currently, european and American main vaccine manufacturers BioNTech and Moderna all adopt the mRNA technology to produce new crown vaccines. One key element in the production of mRNA vaccines is how to deliver mRNA drugs into the body, and the current mature approach is to encapsulate mRNA drugs by liposome nanoparticles for delivery purposes. In recent years, the widely adopted microfluidic technology is used for preparing liposome nano particles with the advantages of high controllability, good reproducibility, small particles and the like. The key of the microfluidic technology is that a microfluidic chip is used, the microfluidic chip is a chip which forms a micro pipeline network on the chip in a micrometer scale space range by utilizing a micro-nano processing technology, and the microfluidic chip is realized by taking controllable fluid as a main operation carrier and miniaturizing basic functions of various laboratories such as biology, chemistry and the like to a chip of a few square centimeters.

Although the problems of nucleic acid medicine preparation are solved to a certain extent by the micro-fluidic technology medicine preparation equipment of the existing brand, most products have the problems of relatively complex system, low automation level, unfriendly product experience and the like, wherein the complex system is represented by: the control system is separated from the preparation instrument, so that the manufacturing cost of the equipment is increased, the instrument is not convenient for a technician to operate, and a large amount of space is occupied in a laboratory.

Low automation levels are expressed in: the control system adopts an independent external computer and software thereof, which is not beneficial to technical personnel to control the instrument.

The product experience is not friendly in that: most of the products on the existing market do not consider the user experience of human-computer interaction, so that the product system does not consider the convenience of users in the use process.

Product compatibility problem: most of the equipment and instruments existing in the market at present are microfluidic chips provided by equipment manufacturers, so that users are limited in selecting different brands and homemade chips.

All the above problems need to be solved.

Disclosure of Invention

The embodiment of the invention aims to provide nano-drug preparation equipment based on a microfluidic technology, which can be used for nano-particle drug preparation, microfluidic drug preparation, microsphere particle preparation and liposome nucleic acid drug preparation.

The embodiment of the invention is realized in such a way that the chip box based on the microfluidic technology comprises liquid inlet holes, liquid outlet holes and chip positions for placing microfluidic chips, wherein the liquid inlet holes and the liquid outlet holes are respectively positioned at two sides of the chip positions, the number of the liquid inlet holes is not less than two, and a circulation pipeline is arranged between the chip positions and the liquid inlet holes and the liquid outlet holes.

Furthermore, the invention also provides nano-drug preparation equipment based on the microfluidic technology, which comprises the chip box, a drug input assembly, a power assembly, a collection assembly and a collection assembly support, wherein the power assembly comprises a first power device and a second power device, the first power device is in driving connection with the drug input assembly, the second power device is in driving connection with the collection assembly support, and the collection assembly is detachably connected with the collection assembly support.

Further, the drug delivery assembly includes syringes in the same number as the cartridge of fluid access holes.

Further, the central axis of the injector and the central axis of the liquid inlet are positioned on the same straight line.

Further, the reagents contained in each of the syringes are different.

Further, the injector is arranged on the injector clamp, the injector clamp is provided with an injector mounting hole and an injector clamping hole, the central axis of the injector clamping hole is perpendicular to the central axis of the injector mounting hole, and the number of the injectors is the same as that of the injector mounting holes.

Further, the device also comprises a control system for controlling the power assembly, wherein the control system is operated through a liquid crystal display screen, and the liquid crystal display screen is arranged on the surface of the equipment shell.

Further, the surface of the equipment shell where the liquid crystal display screen is located is arranged at an included angle with the horizontal plane, and the included angle is an acute angle.

Further, the device housing surface is provided with a recessed area, the cartridge, the drug infusion assembly and the collection assembly support are disposed in the recessed area, the recessed area is covered by a device door, and the device door is rotatably connected with the device housing.

The beneficial effects of the invention are as follows:

1) The technical staff can directly control the power assembly through integrating the liquid crystal display screen outside the instrument, directly input the parameter data of RNA and LNP (Lipid Nanoparticle ) stock solution in the liquid crystal display screen, can start the experiment, need not upper computer such as independent peripheral computer, greatly reduced the experiment flow, shortened the experiment time, improved the automation level, also make equipment more retrench, convenient operation. Meanwhile, technicians can observe the liquid inlet system and control the liquid crystal display at the same time, so that errors can be effectively reduced. Meanwhile, the liquid crystal screen is arranged in the equipment, so that the equipment cost is reduced compared with the form of a peripheral computer.

2) According to the human engineering principle, the product adopts the design which is more in line with man-machine interaction, and the display screen is creatively placed at the top of the instrument, so that the operation of technicians is facilitated, fatigue and damage caused by long-time use of the instrument by the human body are avoided, and the man-machine interaction process is more convenient.

3) The new development of the universal chip conversion box enables users to freely switch different kinds of chip structures, and the compatibility of products is achieved.

4) The new design of the injector clamp can accurately align the injector and ensure the stable flow rate and accuracy.

Drawings

Fig. 1 is an overall structure diagram of a nano-drug preparation device based on a microfluidic technology;

FIG. 2 is an exploded structural view of the nano-drug preparation apparatus provided by the present invention;

FIG. 3 is an internal structural view of an instrument part of the nano-drug preparation apparatus provided by the present invention;

FIG. 4 is a schematic view of the inside of another apparatus part of the nano-drug preparation device provided by the present invention;

FIG. 5 is a block diagram of the rear housing of the apparatus for nano-drug preparation provided by the present invention;

FIG. 6 is an enlarged view of an interior portion of a nano-drug preparation apparatus provided by the present invention;

FIG. 7 is a diagram of a syringe clamp of a nano-drug preparation apparatus provided by the present invention;

FIG. 8 is a test tube fixture diagram of the nano-drug preparation apparatus provided by the present invention;

FIG. 9 is a cartridge holder diagram of the nano-drug preparation apparatus provided by the present invention;

FIG. 10 is a block diagram of a cartridge of the nano-drug preparation apparatus provided by the present invention;

FIG. 11 is a bar graph of the effect of different LNP formulations on the nanoparticles produced;

FIG. 12 is a bar graph of the effect of total flow rate on particle size and uniformity;

FIG. 13 is a bar graph of flow rate versus particle size and uniformity.

Detailed Description

In order to make the technical problems, technical schemes and beneficial effects to be solved more clear, the invention is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.

The invention provides nano-drug preparation equipment based on a microfluidic technology, in particular to nucleic acid liposome preparation equipment developed based on the microfluidic technology. The device is characterized in that two paths of power devices respectively push an injector filled with LNP and nucleic acid medicines to be injected into a microfluidic chip, the LNP and the nucleic acid medicines are mixed through a pipeline in the microfluidic chip, and finally the LNP and the nucleic acid medicines flow into a collected test tube through one path of output pipeline. The device comprises a shell component, a supporting component, a medicine input component, a chip component, a power component, a collecting component and a collecting component supporting frame 7, as shown in fig. 1. The support assembly is connected with the housing assembly, the drug input assembly is connected with the chip assembly, and the power assembly controls the drug input assembly and the collection assembly support 7, and further controls the collection assembly.

Specifically, as shown in fig. 1 to 4, the housing assembly includes the apparatus door 11 and the apparatus housing 12, the support assembly includes the structural support 21, the controller support 22, the driver support 23, the cartridge support 24, the power assembly support 25, the power supply support 26, the structural reinforcement support 27 and the base support 28, the medicine input assembly includes the syringe base 31, the syringe 32 and the syringe clamp 33, the cartridge assembly includes the cartridge case 41 and the cartridge cover 42, the power assembly includes the first power device 51 and the second power device 52, the collection assembly includes the waste liquid test tube and the finished test tube, and the collection assembly support 7 is a test tube rack.

Further, as shown in fig. 1 and 2, the device housing 12 is a box structure, hollow in the middle, and is used for placing a supporting component, a power component, a battery and the like, and is provided with a concave area on the surface, and is used for arranging a medicine input component, a chip component and a collecting component supporting piece 7, which is equivalent to the operation area of the device; as shown in fig. 5, the back surface of the apparatus case 12 is provided with a barrier-shaped vent 121 and a switch hole 122, the vent 121 being used for heat dissipation.

Further, a coupling hole is provided at the upper edge of the concave region of the apparatus housing 12 for coupling with the apparatus door 11, and the apparatus door 11 covers the entire operation region and rotates along the upper edge. Specifically, the equipment door 11 is opened by lifting the handle upwards through the bottom notch or on the door, and is opened and closed at the top with the equipment shell 12 through the connecting hinge, and the door opening angle is more than 180 degrees, so that the operation is convenient in the interior when the door is opened, and meanwhile, the laboratory space is not occupied.

Further, as shown in fig. 3 and 4, the structural support 21 is vertically disposed in the equipment casing 12, and serves as a main equipment body support, and serves as a stabilizing structure and support, and other subsystems are rigidly connected to the structure. Specifically, the controller support 22 is connected to the structural support 21 and the driver support 23, the power assembly support 25 is connected to the structural support 21 for supporting the power assembly, the power source support 26 is for placing the battery, the structural reinforcement support 26 and the base support 28 are connected to the structural support 21, and the base support 28 is rigidly connected to the equipment housing 12.

In other embodiments, the battery may be replaced by an external power source, and correspondingly, the power source support 26 may be omitted.

Further, as shown in fig. 1 and 4, the syringe base 31 has a flat plate structure, the side surface is connected with the first power device 51 through the syringe base connector 311 (in fig. 6), the upper surface is connected with the piston of the syringe 32 in a contact manner, the syringe base 31 is driven by the first power device 51 to move in the vertical direction, specifically, the syringe base 31 pushes the piston of the syringe 32 to move when moving upwards, so that the liquid medicine inside the syringe flows into the chip box 41 to be mixed.

Further, the number of the syringe base 31, the syringe 32 and the first power unit 51 is two, and are connected in one-to-one correspondence.

Further, the solution in the two syringes 32 may be an organic solution containing liposomal LNP particles, and the other may be a diluted solution containing aqueous phase, saline and ethanol, while the mRNA, siRNA nucleic acid drug may be placed in the solution, and the nanoparticle drug encapsulating the nucleic acid drug may be formed after mixing through the microfluidic chip. In this embodiment, two syringes 32 are filled with RNA drug and LNP solution, respectively, and both syringes 32 are simultaneously injected into the cartridge 41.

Further, as shown in fig. 3 and 4, the syringe clamps 33 (in fig. 1) are fixedly connected to the structural brackets 21 by bolts, and each syringe clamp 33 is connected to two syringes 32, specifically, the syringe clamps 33 are connected to the cylinder portions of the syringes 32. The number of the syringe clamps 33 is two, and the syringes are arranged in parallel up and down, so that the connection between the clamped syringes 32 and the bottom surface of the chip box 41 is kept vertical, the flow rate of the syringes is accurately controlled, if the syringes 32 are not fixed by the syringe clamps 33, the power device is directly used for pushing the pistons of the syringes 32 to inject, the syringes 32 are easily inclined, and the flow rate of liquid is further deviated. As shown in fig. 7, the syringe clamp 33 has a plate shape, two syringe clamping holes 331 are formed in a direction perpendicular to the bottom surface of the clamp, and screw holes 332 are formed through the side surface of each of the clamping holes, the screw holes 332 being used for locking the syringe 32. Further, the plate between the two clamping holes 331 is hollowed out, so that a user can conveniently screw in the screw. The injector clamp has a simple structure and is easy to operate, and various types of injectors (1 ml,2ml,5ml,10ml and 20 ml) can be matched for use only by adjusting the distance between the screw and the outer diameter of the injector.

Further, as shown in fig. 6, 9 and 10, the cartridge 41 of the chip assembly is mounted on the cartridge fixing hole 241 of the cartridge holder 24, and the cartridge holder 24 is fixedly coupled to the structural holder 21 by bolts (fig. 4). As shown in fig. 9, two liquid inlets 242 and one liquid outlet 243 are provided on the cartridge holder 24, and the liquid inlets are aligned with the injector 32. As shown in fig. 10, the chip box 41 has a plate-shaped structure, and is provided with a liquid inlet hole, a liquid outlet hole and a chip position 411, wherein the chip position is used for placing a microfluidic chip, and the liquid inlet hole and the liquid outlet hole are respectively positioned at two sides of the chip position 411, so that two paths of liquid medicines of the injector flow into the chip box 41 from the liquid inlet hole and then enter the chip of the chip position 411 for mixing, and flow out from the liquid outlet hole after mixing is completed.

Further, a circulation pipeline is arranged between the chip position and the liquid inlet and outlet.

Further, the liquid inlet and outlet openings of the chip box 41 are in chamfer structures.

Further, the positions of the liquid inlet holes and the liquid outlet holes on the chip box bracket 24 correspond to one of the chip boxes 41.

Further, the chip box cover 42 is used in cooperation with the chip box 41 for sealing the microfluidic chip, and the chip box 42 is fixedly arranged on the chip box bracket 24.

Further, in the present application, when the chip box 41 is used, the chip is only required to be put into the chip box 41, and then the chip box 41 is placed on the chip box bracket 24, meanwhile, the connector inside the chip box 41 can be directly connected with the injector 32, and the chip box 41 can clamp the microfluidic chip and close the microfluidic chip through the pressing plate and the cover plate. The microfluidic chip box designed by the application can be compatible with microfluidic chips made of various materials such as PDMS, glass, COC, COP and stainless steel, and can be compatible with microfluidic chips with any shape within a certain size such as fishbone SHM, Y-type, T-type, tesla structures and the like.

Further, as shown in fig. 4 and 8, the collecting assembly support 7 has an L-shaped structure, and one end of the collecting assembly support 7 is connected with the second power device 52 through a bolt, so that the second power device can control the movement of the collecting assembly support 7, and two test tube slots are formed at the other end of the collecting assembly support 7, including a first slot 61 for loading a waste liquid test tube and a second slot 62 for loading a finished test tube. In other embodiments, multiple test tube slots may be provided and the first and second slot sequences may be reversed.

Further, the power assembly comprises two sets of power devices, each set of power devices respectively and independently operates without interference, the first power device 51 controls the injector 32 to provide constant flow rate for the RNA and LNP liquid inlet pipelines, and the second power device 52 is connected with the collecting assembly supporting frame 7 and is used for horizontally moving the test tube on the supporting frame 7 to the position below the liquid outlet hole of the chip box 42 to receive liquid. Specifically, when the effluent liquid is the prepared nucleic acid solution, the finished product test tube is moved to a liquid outlet for receiving liquid, and when the effluent liquid flows out, the liquid is connected with the liquid by a liquid waste test tube. After the experiment is completed, the support frame 7 automatically returns to the original position, and the second power device 52 realizes the switching of waste liquid and sample receiving.

Further, as shown in fig. 1, the device further comprises a control system and a liquid crystal display screen 8, wherein the control system is used for controlling the device, the control system is integrated inside the instrument, two sets of power devices can be controlled simultaneously, and an operator can input corresponding parameter requirements for the control system through the liquid crystal display screen 8 integrated outside the instrument, so that the power assembly 5 is controlled to finish the preparation of the nanoparticle medicine.

Furthermore, the liquid crystal display screen has a certain inclination angle, accords with the design of human engineering, and is convenient for technicians to input parameters through an operation panel.

The working process of the equipment is as follows: firstly, the second power device drives the waste liquid test tube to run to the liquid outlet hole of the chip box to wait for receiving waste liquid, then the first power device pushes the solution inside the injector to enter the microfluidic chip for mixing, after the waste liquid test tube receives waste liquid, the first power device rapidly moves the finished product test tube to the position below the liquid outlet hole of the microfluidic chip bracket to receive target solution, and when the solution is rapidly connected, the second power device can move the waste liquid test tube again to the position below the liquid outlet hole of the microfluidic chip to receive residual solution. Finally, the second power device moves the waste liquid test tube and the liquid receiving test tube to return to the initial positions. The input of control parameters in the process is completed by a liquid crystal display screen arranged on the equipment.

Three verification experiments of LNP prescriptions were performed using the nanoparticle preparation apparatus developed by the product of this patent, and the results are shown in Table 1.

Table 1 comparison of the results of the preparation

The lipid particle size obtained by three groups of experiments is between 50 and 65 nanometers, the particle uniformity parameter PDI is smaller than 0.1, and the particle quality is very good.

Meanwhile, four comparative experiments were performed by using the third prescription through the nanoparticle preparation apparatus developed by the product of this patent, and the results are shown in table 2.

Table 2 comparison of four sets of experimental results

Four sets of experiments compare the effect of different total flow rates and different flow rate ratios on liposome nanoparticles. As can be seen from fig. 12, the lipid particle size obtained is between 50 and 75 nm with a flow rate ratio of greater than 3, the particle uniformity parameter PDI is less than 0.1, and the particle size is greater than 100nm with a flow rate ratio of less than 3. Meanwhile, the particle size and uniformity of the total flow rate of 6ml/min are better than those of the total flow rate of 8.5ml/min and 10 ml/min. This result is highly consistent with the relevant literature and empirical verification results. The effectiveness and the accuracy of the nanoparticle preparation equipment provided by the application can be proved.

FIGS. 11-13 show the effect of different formulations, total flow rates and flow rate ratios on nanoparticle size and uniformity, highly consistent with results in scientific literature, and highly reliable results, demonstrating that the nanoparticle preparation system provided by the application can meet the requirements for preparing nanoparticle medicaments.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the invention, but any modifications, equivalents, improvements, etc. within the principles of the present invention should be included in the scope of the present invention.

Claims (9)

1. The utility model provides a chip box based on micro-fluidic technology, its characterized in that includes feed liquor hole, goes out the liquid hole and is used for placing the chip position of micro-fluidic chip, the feed liquor hole with go out the liquid hole and be located respectively the both sides of chip position, feed liquor hole quantity is not less than two, the chip position with the feed liquor hole with go out and be provided with the circulation pipeline between the liquid hole.

2. The nano-drug preparation device based on the microfluidic technology, which is characterized by comprising the chip box as claimed in claim 1, and further comprising a drug input assembly, a power assembly, a collection assembly and a collection assembly support, wherein the power assembly comprises a first power device and a second power device, the first power device is in driving connection with the drug input assembly, the second power device is in driving connection with the collection assembly support, and the collection assembly is detachably connected with the collection assembly support.

3. The nano-drug preparation apparatus of claim 2, wherein the drug input assembly comprises a number of syringes equal to the number of access holes of the chip cartridge.

4. A nano-drug preparation apparatus as in claim 3, wherein the central axis of the syringe and the central axis of the inlet are collinear.

5. A nano-drug preparation apparatus as in claim 3, wherein the reagents contained in each of said syringes are different.

6. A nano-drug preparation apparatus as claimed in claim 3, wherein the syringes are provided on a syringe clamp provided with syringe mounting holes and syringe clamping holes, the syringe clamping holes having a central axis perpendicular to the syringe mounting hole central axis, the number of syringes being the same as the number of syringe mounting holes.

7. The nano-drug preparation device of claim 2, further comprising a control system for controlling the power assembly, the control system being operated by a liquid crystal display, the liquid crystal display being disposed on a surface of a housing of the device.

8. The nano-drug preparation device according to claim 7, wherein the surface of the device housing where the liquid crystal display is located is disposed at an angle with respect to the horizontal plane, and the angle is an acute angle.

9. The nano-drug preparation device of claim 7, wherein the device housing surface is provided with a recessed area, the cartridge, the drug input assembly and the collection assembly support being disposed within the recessed area, the recessed area being covered by a device door rotatably coupled to the device housing.