Method and apparatus for synthesizing liposomes

Abstract

The application provides a liposome synthesis method and a synthesis device, and the method comprises the following steps: two liquids of an organic phase and an aqueous phase are respectively introduced into two sample introduction channels of a Y-shaped flow channel; the Y-shaped flow channel comprises the two sample introduction channels and a synthesis channel which is in communication with the converging position of the two sample introduction channels; a special ultrasonic device in the shape of a regular pentagon arranged at the bottom of the synthesis channel is driven to work, and at least through the micro-vortices generated in the liquid in the synthesis channel and distributed at the positions of the edges of the regular pentagon during the working of the special ultrasonic device, the two liquids in the synthesis channel are mixed to generate target liposomes. Through the cooperative design of the micro-flow channel structure and the special ultrasonic device, the synthesis efficiency of the liposomes can be significantly improved, and the average particle size and uniformity of the product in the synthesis process of the liposomes are optimized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine and molecular biology, and particularly relates to a liposome synthesis method and a synthesis device. BACKGROUND

[0002] Liposomes (or lipid globules, liquid crystal microcapsules) have a cell-like membrane structure with lipids such as phospholipids and cholesterol as membrane materials, and can be used as drug carriers to achieve drug delivery, thereby effectively solving the problems of poor bioavailability, low plasma solubility, and high clearance rate of free drugs. However, the drug delivery effect and pharmacokinetic circulation half-life are strongly affected by the size of the liposomes. Some studies have shown that liposomes larger than 200 nm can cause serious toxicity, and liposomes smaller than 50 nm can accumulate in the liver, while liposomes with a size of about 100 nm can maintain good stability in blood circulation, which makes them exhibit better biodistribution characteristics in vivo. Therefore, during the synthesis of liposomes, accurately controlling the size and uniformity of the liposomes is crucial for optimizing their drug properties.

[0003] Microfluidic technology is a technology for precisely controlling and manipulating microscale fluids, which can be applied in technical fields such as medicine, biotechnology, and chemical industry. For example, two or more liquids can be mixed at high speed by microfluidic technology to perform a rapid reaction to generate liposomes.

[0004] Currently, when liposomes are synthesized by microfluidic technology, the control of the size of the liposomes highly depends on the flow rate, and there are problems such as long mixing distance and long mixing time. Therefore, how to effectively control the size (particle size and uniformity) of the liposomes to generate liposomes with controllable particle size and uniform size is a technical problem to be solved. SUMMARY

[0005] Therefore, the present application provides a liposome synthesis method and a synthesis device, which can significantly improve the synthesis efficiency of liposomes and optimize the average particle size and uniformity of the product in the synthesis process of liposomes through the collaborative design of microfluidic structures and special ultrasonic devices.

[0006] The first aspect of the present application provides a liposome synthesis method, comprising:

[0007] Two liquids of an organic phase and an aqueous phase are respectively introduced into two sample introduction channels of a Y-shaped flow channel; the Y-shaped flow channel comprises the two sample introduction channels and a synthesis channel in communication with the convergence of the two sample introduction channels;

[0008] The pentagonal special ultrasonic device arranged at the bottom of the synthesis channel is driven to work, and at least by the micro-vortices generated in the liquid in the synthesis channel during the working of the pentagonal special ultrasonic device, the two liquids in the synthesis channel are mixed to generate target liposomes.

[0009] From the above, in the liposome synthesis method provided by the application, the organic phase and the aqueous phase are respectively poured into the two sample introduction channels of the Y-shaped flow channel, so that the two liquids converge into the synthesis channel of the Y-shaped flow channel, and then the pentagonal special ultrasonic device is arranged at the bottom of the synthesis channel and is driven to work. By using the micro-vortices generated during the working of the pentagonal special ultrasonic device and distributed at the positions of the edges of the pentagon, local strong disturbance and flow field change are generated to promote molecular diffusion and convective mixing between the liquids, so that the mixing effect of the organic phase and the aqueous phase in the synthesis channel can be significantly enhanced, and target liposomes are generated. Moreover, the design of the Y-shaped flow channel enables the two liquids to be poured into the synthesis channel according to a predetermined ratio and flow rate, and the working parameters (such as frequency and power) of the special ultrasonic device can be accurately adjusted, so that the mixing process can be accurately controlled, and the synthesis efficiency of liposomes of different sizes can be improved.

[0010] Optionally, the special ultrasonic device is arranged at the convergence of the two sample introduction channels and the synthesis channel, wherein two adjacent edges of the pentagonal special ultrasonic device are respectively directed towards the two sample introduction channels.

[0011] From the above, the convergence of the two sample introduction channels and the synthesis channel is the place where the liquids of the two sample introduction channels converge. At the convergence, the liquids of the two sample introduction channels collide with each other at a certain angle and speed. By arranging the special ultrasonic device at the convergence, the liquids can be mixed as soon as they contact and begin to converge. The momentum and energy of the liquids during the convergence can be fully utilized, and the mixing process can be more efficient in combination with the action of the micro-vortices.

[0012] Optionally, the special ultrasonic device has a hollowed-out pentagonal ring structure, and an outer vertex of the pentagonal ring structure is located on the intersection line of the two liquids poured into the synthesis channel.

[0013] At least by the micro-vortices generated in the liquid in the synthesis channel during the working of the special ultrasonic device and distributed at the positions of the inner and outer edges of the pentagonal ring structure, the two liquids in the synthesis channel are mixed to generate target liposomes.

[0014] Optionally, the special ultrasonic device has a hollowed-out pentagonal ring structure, and an outer vertex of the pentagonal ring structure is located on the intersection line of the two liquids poured into the synthesis channel. A second pentagonal shape or a second pentagonal ring structure is further nested in the pentagonal ring structure.

[0015] At least through the working of the special ultrasonic device, the micro-vortices generated in the liquid in the synthesis channel and distributed at the positions of the inner and outer edges of the regular pentagonal ring, the positions of the edges of the nested second regular pentagon, or the positions of the inner and outer edges of the second regular pentagonal ring mix the two liquids in the synthesis channel to generate target liposomes.

[0016] Therefore, the shape of the special ultrasonic device can adopt a regular pentagonal ring structure, or a smaller regular pentagonal structure or a regular pentagonal ring structure is nested in the regular pentagonal ring structure, which can form a multi-stage acoustic field distribution to act on a larger range in the synthesis channel, generate more micro-vortices, have more acoustic fluid tunnels, and have more mixing areas or equivalent longer acoustic fluid tunnels when applied to the mixing of different liquids or different particles in the liquid in the convection channel or the mixing-based reaction, so that the mixing effect is better.

[0017] In a second aspect, the application provides a liposome synthesis device for implementing the liposome synthesis method described above, which comprises:

[0018] A Y-shaped flow channel, which comprises two sample introduction channels and a synthesis channel connected to the convergence of the two sample introduction channels, and the two sample introduction channels are used to introduce two liquids of organic phase and aqueous phase;

[0019] A regular pentagonal special ultrasonic device arranged at the bottom of the synthesis channel, which is used to generate micro-vortices in the liquid in the synthesis channel and distributed at the positions of the edges of the regular pentagon when working, so as to mix the two liquids in the synthesis channel to generate target liposomes.

[0020] Optionally, the special ultrasonic device is arranged at the convergence of the two sample introduction channels and the synthesis channel, and adjacent two edges of the regular pentagonal special ultrasonic device are respectively directed to the two sample introduction channels.

[0021] Optionally, the shape of the special ultrasonic device forms a regular pentagonal ring structure with an internal hollow, and an outer vertex of the regular pentagonal ring structure is located on the intersection line of the two liquids introduced into the synthesis channel.

[0022] At least through the working of the special ultrasonic device, the micro-vortices generated in the liquid in the synthesis channel and distributed at the positions of the inner and outer edges of the regular pentagonal ring, the positions of the edges of the nested second regular pentagon, or the positions of the inner and outer edges of the second regular pentagonal ring mix the two liquids in the synthesis channel to generate target liposomes.

[0023] Optionally, the shape of the special ultrasonic device forms a regular pentagonal ring structure with an internal hollow, and an outer vertex of the regular pentagonal ring structure is located on the intersection line of the two liquids introduced into the synthesis channel. The regular pentagonal ring structure further nests a second regular pentagon or a second regular pentagonal ring structure.

[0024] At least by the operation of the special ultrasonic device, the micro-vortices distributed at the positions of the inner and outer edges of the first regular pentagonal ring, the positions of the edges of the nested second regular pentagonal ring, or the positions of the inner and outer edges of the second regular pentagonal ring, which are generated in the liquid in the synthetic channel, mix the two liquids in the synthetic channel to generate target liposomes.

[0025] Optionally, the synthetic channel has a linear shape, a width of 100 microns-160 microns, and a length of 500 microns-6 millimeters.

[0026] As described above, the linear channel has a simple structure, and the flow rate distribution of the liquid in the linear channel is relatively uniform, which helps to ensure that the mixing process of the liquid in the channel is relatively stable and reduces fluctuations in the mixing effect caused by uneven flow rate.

[0027] Optionally, the special ultrasonic device adopts a continuous operation mode or a pulse operation mode, and has a power of 250 milliwatts-400 milliwatts.

[0028] The flow rate ratio of the organic phase to the aqueous phase is 1:1-1:9.

[0029] As described above, in the continuous operation mode, the special ultrasonic device continuously generates sound waves, which can form a stable sound field environment in the synthetic channel. The continuous sound wave action can generate a continuous driving force for the liquid, which helps to stably control the flow rate and direction of the liquid in the synthetic channel. In the pulse operation mode, the action time and interval of the sound wave can be accurately controlled according to the needs, and special flow field effects such as transient vortex and jet can be generated. These special flow fields can be used for some special microfluidic operations, which opens up a new application field for microfluidic technology. In addition, by adjusting the flow rate ratio of the organic phase to the aqueous phase, the mixing degree can be accurately controlled. For example, when the flow rate ratio is close to 1:1, the amount of the two-phase liquid is relatively balanced, the sound wave can be uniformly dispersed in the two phases, and a relatively stable mixing system is formed. When the flow rate ratio is 1:9, the water phase accounts for a large proportion, and the sound wave can more effectively disperse the organic phase in the large amount of water phase, so as to realize the uniform emulsification or dissolution of the organic phase in the water phase.

[0030] These and other aspects of the application will become more apparent from the following description of the (multiple) embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1a is a schematic diagram of the jet phenomenon and the secondary flow phenomenon generated by the special ultrasonic device provided by the embodiments of the application;

[0032] Figure 1b is a schematic diagram of the acoustic fluid tunnel provided by the embodiments of the application;

[0033] Figure 2 A flow chart of a method for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 1.

[0034] Figure 3a A schematic diagram of a first implementation of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 2.

[0035] Figure 3b A schematic diagram of a second implementation of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 3.

[0036] Figure 3c A schematic diagram of a third implementation of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 4.

[0037] Figure 4a A schematic diagram of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 5. Figure 3a A schematic diagram of a synthesis simulation result of the device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 6.

[0038] Figure 4b A schematic diagram of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 7. Figure 3b A schematic diagram of a synthesis simulation result of the device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 8.

[0039] Figure 4c A schematic diagram of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 9. Figure 3c A schematic diagram of a synthesis simulation result of the device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 10.

[0040] Figure 5 A schematic diagram of a device for synthesizing a liposome according to an embodiment of the present application is shown in FIG. 11. Figures 4a-4c A schematic diagram of a mixing frequency comparison of a synthesis simulation according to an embodiment of the present application is shown in FIG. 12.

[0041] It should be understood that the size and shape of each block in the above structural schematic diagram are only for reference, and should not constitute an exclusive interpretation of the embodiments of the present application. The relative position and inclusion relationship between the blocks presented by the structural schematic diagram are only used to represent the structural association between the blocks, and do not limit the physical connection mode of the embodiments of the present application. DETAILED DESCRIPTION

[0042] The technical solutions provided by the present application will be further described below in conjunction with the drawings and embodiments. It should be understood that the system structure and business scenarios provided in the embodiments of the present application are mainly to illustrate possible implementation modes of the technical solutions of the present application, and should not be interpreted as the only limitation of the technical solutions of the present application. Those skilled in the art can know that the technical solutions provided by the present application are also applicable to similar technical problems as the system structure evolves and new business scenarios appear.

[0043] It should be understood that the liposome synthesis schemes provided in this application include liposome synthesis apparatus and liposome synthesis methods, as well as their applications. Since these technical solutions solve problems based on the same or similar principles, some repetitions may not be repeated in the following descriptions of specific embodiments, but these specific embodiments should be considered as mutually referencing each other and can be combined with each other.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. In case of any inconsistency, the meaning set forth in this specification or derived from the content described herein shall prevail. Furthermore, the terminology used herein is for the purpose of describing embodiments of this application only and is not intended to limit this application. To accurately describe the technical content of this application and to accurately understand the invention, the following explanations or definitions of the terms used in this specification are provided before describing specific embodiments:

[0045] 1) Ultrasonic Device: A high-frequency resonator, which can be a device that generates mechanical vibration by applying voltage based on the piezoelectric effect. In this application, a piezoelectric resonator that generates ultrasonic waves of not less than 0.5 GHz (GHz is equivalent to gigahertz) during operation is used. Preferably, it is a piezoelectric resonator that generates ultrasonic waves of not less than 1 GHz and not more than 30 GHz during operation, for example, it can be 2 GHz to 2.5 GHz. Such piezoelectric resonators can be, for example, surface acoustic wave (SAW) devices, bulk acoustic wave (BAW) devices, etc. For example, when it is BAW, it can be a thin-film bulk acoustic wave resonator (FBAR), a solid-state assembled resonator (SMR), or a Lamb wave resonator (LWR). For ease of description, the piezoelectric resonator that can generate ultrasonic waves of not less than 0.5 GHz will be referred to as an ultrasonic device from now on.

[0046] 2) Jet phenomenon: This is a phenomenon that occurs when the sound waves from an ultrasonic device act on a liquid. The regional vibration generated at the working interface of the ultrasonic device can form a traveling wave in the liquid and exert a continuous thrust on the local liquid in the liquid environment, causing at least a portion of the liquid to move in a straight line along the direction of sound wave propagation. This phenomenon of straight-line movement is called jet phenomenon.

[0047] Secondary flow phenomena, including eddies and thermal backflow, are another phenomenon generated when ultrasonic devices act on liquids. They include eddies (or micro vortices) caused by local circulation generated by the jet driving the liquid, and thermal backflow generated by the heating of ultrasonic devices.

[0048] The jet phenomenon and secondary flow phenomenon can be found in [reference needed]. Figure 1aThe images and diagrams shown. Figure 1a The diagram shows a schematic of using eddies (i.e., microvortices) to capture particles.

[0049] 3) Acoustic-fluid tunnel: Under the excitation of an input signal, the ultrasonic device generates ultra-high frequency vibrations emitting sound waves. These sound waves propagate through the fluid, inducing directional movement of the fluid (jet phenomenon), further forming micro-vortices. One such phenomenon is... Figure 1b The simulation diagram shows that several tiny fluid microvortices (such as...) can be generated at the edge of the chip in the ultrasonic device. Figure 1a As shown in the diagram, these fluid microvortices can trap particles in the liquid. Combined with the positional distribution of these fluid microvortices, they exhibit a phenomenon where the particles in the liquid are distributed along the edge of the ultrasonic device chip. Since the particles in the fluid flow through these fluid microvortices distributed along the chip edge, the paths that these fluid microvortices create for the particles to flow through (such as...) Figure 1b As shown in the figure, this application refers to it as an acoustic fluid tunnel.

[0050] 4) Shape of the ultrasonic device: for example, pentagonal, leaf-shaped, spindle-shaped, etc. In this embodiment, the shape of the ultrasonic device refers to the shape of its resonant region. The resonant region is formed by the overlapping regions of the orthographic projections of the bottom electrode layer, piezoelectric layer, and top electrode layer (these three layers can be called resonant layers) onto the substrate. For example, one structure of the resonant layer of the pentagonal ultrasonic device can be composed of a pentagonal bottom electrode layer, piezoelectric layer, and top electrode layer of the same shape with overlapping orthographic projections. Another structure can be composed of bottom electrode layers, piezoelectric layers, and top electrode layers of different shapes, such as a quadrilateral bottom electrode layer, with the projections of the piezoelectric layer and top electrode layer covering the quadrilateral being a pentagon located within the quadrilateral. Thus, the overlapping regions of the orthographic projections of the bottom electrode layer, piezoelectric layer, and top electrode layer onto the substrate are the pentagon, forming the pentagonal ultrasonic device. The term "ultrasonic device" is used here only to illustrate the definition of the shape of the ultrasonic device in this application and does not limit the ultrasonic device to include other layered structures. For example, a reflective layer (such as a Bragg reflective layer, a cavity reflective layer, etc.) may be disposed below the resonant layer, and aluminum nitride may be disposed above the resonant layer as a protective layer. Similarly, unless otherwise specified, the area of ​​the ultrasonic device mentioned in this application refers to the area of ​​the resonant region of the ultrasonic device.

[0051] For example, a supersonic device with a regular pentagonal ring structure can have a top electrode that is a regular pentagonal ring, a bottom electrode layer and a piezoelectric layer that are pentagonal, and a resonant region formed by projection that is a regular pentagonal ring.

[0052] For example, a regular pentagonal ring structure can be nested within a second regular pentagon. This can be achieved by setting the top electrode as a regular pentagonal ring structure nested within the second regular pentagon, with the top electrode's regular pentagonal ring structure electrically connected to the second regular pentagon.Figure 3c The electrical connection point is not shown in the diagram; the electrical connection between the regular pentagonal ring structure and the second regular pentagon has little impact on the overall mixing. Alternatively, the electrical connection point between the bottom electrode and the top electrode can be hollowed out, so that the projected resonant region does not include the electrical connection point (matching). Figure 3c (The shape in the middle).

[0053] The term "ultrasonic device" is used here only to illustrate the definition of the shape of the ultrasonic device in this application and does not limit the ultrasonic device to include other layered structures. For example, a reflective layer (such as a Bragg reflective layer, a cavity reflective layer, etc.) may be disposed below the resonant layer, and aluminum nitride may be disposed above the resonant layer as a protective layer. Similarly, unless otherwise specified, the area of ​​the ultrasonic device mentioned in this application refers to the area of ​​the resonant region of the ultrasonic device.

[0054] The solutions provided in this application will be described in detail below with reference to the accompanying drawings and embodiments.

[0055] This application proposes a method for synthesizing liposomes and a liposome synthesis apparatus for implementing the method. Through the synergistic design of microfluidic structure and ultrasonic device, the synthesis efficiency of liposomes can be significantly improved, and the average particle size and uniformity of the products during the synthesis process can be optimized.

[0056] like Figure 2 As shown in the embodiments of this application, a method for synthesizing liposomes is provided, the method comprising:

[0057] S110: The two liquids, organic phase and aqueous phase, are respectively introduced into the two injection channels of the Y-shaped flow channel;

[0058] S120: Drive the pentagonal ultrasonic device located at the bottom of the synthesis channel of the Y-shaped flow channel to work, at least through the micro vortices generated in the liquid in the synthesis channel and distributed at the positions of each side of the pentagon when it is working, to mix the two liquids in the synthesis channel to generate target liposomes.

[0059] In this embodiment, an organic phase and an aqueous phase are introduced into two injection channels of a Y-shaped flow channel, respectively, so that the two liquids converge into the synthesis channel of the Y-shaped flow channel. Then, by setting a regular pentagonal ultrasonic device at the bottom of the synthesis channel and driving it to work, the micro vortices generated by the device and distributed on each side of the regular pentagon can generate strong local disturbances and flow field changes, promoting molecular diffusion and convection mixing between the liquids. This can significantly enhance the mixing effect of the organic phase and the aqueous phase in the synthesis channel, thereby generating the target liposomes.

[0060] In some embodiments, the height of the synthesis channel also affects the mixing results. For example, reducing the channel height from 140 μm to 40 μm further reduces the particle size under the same power and pulse conditions, resulting in an average particle size of 110 nm. At a low channel height (40 μm), the effect of different pulse conditions on particle size generation is more significant. A high channel height (140 μm) is compatible with pulsed operating modes at 1 W and 2 W; while at a low channel height (40 μm), the optimal pulsed operating mode is at 1 W.

[0061] Reference Figure 3a As shown in the embodiments of this application, a liposome synthesis apparatus includes:

[0062] The Y-shaped flow channel has two injection channels and a synthesis channel connected to the confluence of the two injection channels. The two injection channels are used to inject two liquids, an organic phase and an aqueous phase.

[0063] A regular pentagonal ultrasound device is disposed at the bottom of the synthesis channel. When it is working, the micro vortices generated in the liquid within the synthesis channel and distributed at the positions of each side of the regular pentagon mix the two liquids in the synthesis channel to generate target liposomes.

[0064] In some embodiments, the synthesis channel is linear in shape, with a width of 100-160 micrometers and a length of 500-6 millimeters. In a linear channel, the liquid flow rate distribution is relatively uniform, which helps to ensure a relatively stable mixing process within the channel and reduces fluctuations in the mixing effect caused by uneven flow rates.

[0065] In some embodiments, the aforementioned ultrasonic device employs either a continuous operating mode or a pulsed operating mode, with a power of 250 milliwatts to 400 milliwatts. In continuous operating mode, the ultrasonic device continuously generates sound waves, creating a stable acoustic field environment within the synthesis channel. The continuous sound waves exert a sustained driving force on the liquid, facilitating stable control of the liquid's flow velocity and direction within the synthesis channel. In pulsed operating mode, the duration and interval of the sound waves can be precisely controlled as needed, generating special flow field effects such as transient vortices and jets. These special flow fields can be used for specific microfluidic operations, such as particle capture, separation, and manipulation, opening up new application areas for microfluidic technology. Experimental simulation results show that, using the same power, the particle size synthesized in continuous operating mode is smaller than or equal to that synthesized in pulsed operating mode. Furthermore, when using pulsed operating mode, for the same pulse duration, a shorter period results in a smaller synthesized particle size; conversely, for the same pulse period, a longer pulse duration results in a smaller synthesized particle size.

[0066] In some embodiments, an organic phase and an aqueous phase are introduced through two injection channels of a Y-shaped flow channel, respectively. The organic phase can be a mixture of DPPC (dispalmitoylphosphatidylcholine) and cholesterol, a mixture of DSPC (distearylphosphatidylcholine) and cholesterol, or POPC (palmitoyloleylphosphatidylcholine). The aqueous phase can be pure water. Then, an acoustic-fluid tunnel corresponding to the contour edge of the acoustic-fluid device is generated in the liquid within the synthesis channel using a special ultrasonic device. The two liquids within the synthesis channel are mixed through this acoustic-fluid tunnel to generate the target liquid (e.g., a liposome solution).

[0067] In some embodiments, such as Figure 3a As shown, the aforementioned regular pentagonal ultrasonic device can be positioned at the convergence of the two sample inlet channels and the synthesis channel, with adjacent sides of the pentagonal ultrasonic device facing the two sample inlet channels respectively. The convergence of the two sample inlet channels and the synthesis channel is where the liquids from the two inlet channels converge. At this convergence point, the liquids from the two inlet channels collide with each other at a certain angle and velocity. By placing the ultrasonic device at this location, the micro-vortices it generates can completely cover the liquid stream entering the synthesis channel, enabling mixing as soon as the liquids come into contact and begin to converge. This fully utilizes the momentum and energy of the liquid during convergence, combined with the effect of the micro-vortices, to create uniform disturbance in the liquid, significantly shortening the mixing start-up time and making the mixing process more efficient. In addition, one of the outer vertices of the regular pentagonal ultrasonic device is located at the interface between the two liquids in the synthesis channel (that is, the regular pentagonal structure is symmetrically set at the interface between the two liquids in the synthesis channel), which allows the acoustic fluid effect generated by the ultrasonic device to be directly focused on the liquid interface region. This precise positioning can effectively break the interfacial tension between the two liquids, promote the mutual penetration and diffusion between molecules, and significantly enhance the mixing effect.

[0068] Generally, the area of ​​an ultrasonic transducer (UTT) is negatively correlated with its resonant frequency (natural frequency); the larger the area, the lower the resonant frequency. However, the resonant frequency of an UTT is positively correlated with its ability to manipulate particles and microfluidics. A higher resonant frequency means a greater mixing capacity for microfluidics with the same driving power. Therefore, smaller UTT devices typically have higher resonant frequencies, generating stronger eddies and achieving greater mixing capabilities for microscale fluids, such as producing smaller particles generated during the mixing process. On the other hand, a smaller UTT area results in a smaller effective range within the flow channel, which is detrimental to mixing microfluidics within a larger space. Furthermore, a smaller UTT area also means a shorter acoustic-fluid tunnel, which is also unfavorable for the limited space available for mixing microfluidics. For example, for mixing high-fluidity, multiphase fluids within a flow channel, the small UTT area may, due to the aforementioned two reasons, result in mixing efficiency that does not meet the requirements for generating nanoscale products.

[0069] Based on this, such as Figure 3b As shown, in some embodiments of this application, the aforementioned ultrasonic device can employ an internally hollowed-out regular pentagonal ring structure, with the internal hollowed-out shape being a regular pentagon. One outer vertex of this regular pentagonal ring structure is located at the interface between the two liquids flowing into the synthesis channel. Alternatively, as... Figure 3c As shown, a smaller regular pentagon or regular pentagonal ring structure can be nested inside the aforementioned hollowed-out regular pentagonal ring structure.

[0070] based on Figures 3b-3c This application, by employing an internally hollowed-out regular pentagonal ring structure or a nested regular pentagonal ring structure, compared to... Figure 3a The pentagonal structure shown has the same total area as the regular pentagonal structure, but its internal hollow or nested pentagonal ring structure can form a multi-level sound field distribution, resulting in a larger effective range within the flow channel and an increase in the number of micro-vortices (with more acoustic-fluid tunnels). It also shows good results when applied to mixing different liquids or in mixing-based reactions. For example, when different liquids flowing from different branches upstream of the flow channel pass through the ultrasonic device provided in this embodiment, the liquid over a larger area, under the action of more micro-vortices, forms more turbulence or flow in space, or passes through longer acoustic-fluid tunnels, thereby achieving a better mixing effect and generating target liposomes with smaller average particle size and better uniformity.

[0071] based on Figures 3a-3c The illustrated liposome synthesis apparatus, along with simulation results of related experiments based on the above-described embodiments of this application, is also provided. For example, refer to... Figures 4a-4c and Figure 5As shown, under the same experimental conditions (same flow ratio, same power 400mW), respectively using Figures 3a-3c The liposome synthesis apparatus shown was used to conduct synthesis experiments on both organic and aqueous liquids to evaluate the effect of ultrasonic devices with different geometries on liquid mixing in the liposome synthesis apparatus, so as to further optimize the average particle size and uniformity (monodispersity coefficient) of the products during the liposome synthesis process.

[0072] in Figure 4a For use Figure 3a A schematic diagram of the simulation results of the synthesis experiment of the liposome synthesis device. The simulation results show that the mixing area is concentrated near the vertex of the pentagon, but the overall mixing uniformity is generally poor. Figure 4b For use Figure 3b A schematic diagram of the simulation results of the synthesis experiment of the liposome synthesis device. The simulation results show that the internally hollowed-out regular pentagonal ring structure helps to expand the range of sound field action and increase the intensity of local disturbance. Figure 4c For use Figure 3c The diagram shows the simulation results of the liposome synthesis device. The simulation results show that the nested structure of 20μm pentagonal rings and small pentagons exhibits the best mixing effect and the highest mixing frequency, indicating that this nested structure can more effectively promote mixing during the synthesis of liposomes and help to achieve the generation of small-diameter liposomes.

[0073] Figure 5 for Figures 4a-4c The diagram shows a comparison of the mixing frequencies in the synthetic simulation. The mixing frequency of the regular pentagonal structure is about 58%, the mixing frequency of the hollowed-out regular pentagonal ring structure is about 82%, and the mixing frequency of the nested structure of pentagonal ring and small pentagon is about 96%, which is the optimal mixing frequency.

[0074] The simulation results provided in this application show that the ultrasonic device with an internally hollowed-out or nested regular pentagonal ring structure provided in the embodiments of this application, compared with the ultrasonic device with a regular pentagonal structure, has a relatively small decrease in resonant frequency due to the unchanged area, thus maintaining a high resonant frequency. However, the overall area formed by the pentagons of the internally hollowed-out or nested structure is larger than that of the regular pentagonal structure, and the number of micro vortices that can be generated increases. When applied to liquid mixing or mixing-based reactions, more acoustic-fluid tunnels can be generated, and the overall flow can form turbulence or turbulence in a larger space, or a longer flow through the acoustic-fluid tunnel, which is beneficial for the mixing or mixing reaction of different liquids or different particles in the liquid.

[0075] In some embodiments, when the organic and aqueous phases flow into the synthesis channel in a certain ratio, the acoustic waves generated by the ultrasonic device can break the interfacial tension between the two phases, allowing the two liquid phases to fully contact and mix. By adjusting the flow ratio of the organic and aqueous phases, the degree of mixing can be precisely controlled. For example, when the flow ratio is close to 1:1, the amounts of the two liquid phases are relatively balanced, and the acoustic waves can be dispersed more evenly in the two phases, forming a relatively stable mixing system. As the flow ratio increases from 1:1 to 1:9, with the same power applied (250mW to 400mW), the particle size tends to decrease. When the flow ratio is 1:3 or 1:4, the water-alcohol interface is located in the middle, and the effective mixing area of ​​the device is the largest. When 400mW is applied, the average particle size decreases to 120nm. However, when the flow ratio is increased to 1:9, the water-alcohol interface deviates from the center, and the minimum particle size reaches 58nm, but impurity peaks appear in the 500-1000nm range, resulting in a large PDI (Polydispersity Index).

[0076] In summary, the liposome synthesis apparatus provided in this application provides a supersonic device at the bottom of the synthesis channel in a Y-shaped flow channel. The device generates microvortices during operation, which produce strong local disturbances and flow field changes, promoting molecular diffusion and convective mixing between liquids. This significantly enhances the mixing effect of the two liquids within the synthesis channel. Furthermore, the Y-shaped flow channel design allows the two liquids to flow into the synthesis channel according to a predetermined ratio and flow rate. The operating parameters of the supersonic device (such as frequency and power) can be precisely adjusted, thereby achieving precise control of the mixing process and improving the mixing efficiency of the liquids.

[0077] It should be noted that the embodiments described in this application are merely some embodiments, not all embodiments. The components of the embodiments of this application typically described and shown in the accompanying drawings can be arranged and designed in various different configurations. Therefore, the above detailed description of the embodiments of this application provided in the accompanying drawings is not intended to limit the scope of the claimed application, but merely represents selected embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

[0078] The terms "first, second, third, etc." or similar terms such as module A, module B, module C, etc., used in the specification and claims are only used to distinguish similar objects and do not represent a specific ordering of objects. It is understood that a specific order or sequence may be interchanged where permitted so that the embodiments of this application described herein can be implemented in an order other than that illustrated or described herein.

[0079] In the above description, the labels indicating the steps do not necessarily mean that the steps will be executed. They may include intermediate steps or be replaced by other steps. Where permissible, the order of the steps may be interchanged or executed simultaneously.

[0080] The term "comprising" as used in the specification and claims should not be construed as limiting itself to what follows; it does not exclude other elements or steps. Therefore, it should be interpreted as specifying the presence of the mentioned feature, integral, step, or component, but does not exclude the presence or addition of one or more other features, integrals, steps, or components, or groups thereof. Thus, the statement "device comprising means A and B" should not be limited to a device consisting solely of components A and B.

[0081] The terms "an embodiment" or "an embodiment" as used in this specification mean that a particular feature, structure, or characteristic described in conjunction with that embodiment is included in at least one embodiment of this application. Therefore, the terms "in one embodiment" or "in an embodiment" appearing throughout this specification do not necessarily refer to the same embodiment, but may refer to the same embodiment. Furthermore, in the various embodiments of this application, unless otherwise specified or in case of logical conflict, the terminology and / or descriptions between different embodiments are consistent and can be mutually referenced. Technical features in different embodiments can be combined to form new embodiments based on their inherent logical relationships.

[0082] Note that the above are merely preferred embodiments and the technical principles employed in this application. Those skilled in the art will understand that the present invention is not limited to the specific embodiments described herein, and various obvious changes, readjustments, and substitutions can be made without departing from the scope of protection of the present invention. Therefore, although the present application has been described in detail through the above embodiments, the present invention is not limited to the above embodiments, and may include many other equivalent embodiments without departing from the concept of the present invention, all of which fall within the scope of protection of the present invention.

Claims

1. A method for synthesizing liposomes, characterized in that, include: Two liquids, an organic phase and an aqueous phase, are respectively introduced into two injection channels of a Y-shaped flow channel; the Y-shaped flow channel includes the two injection channels and a synthesis channel connected to the confluence of the two injection channels. The device drives a regular pentagonal ultrasonic device located at the bottom of the synthesis channel. This ultrasonic device is positioned at the confluence of the two sample inlet channels and the synthesis channel, with adjacent sides of the regular pentagonal ultrasonic device facing the two sample inlet channels. The ultrasonic device is shaped to form an internally hollowed-out regular pentagonal annular structure, with one outer vertex of the regular pentagonal annular structure located at the boundary line between the two liquids flowing into the synthesis channel. A second regular pentagon or a second regular pentagonal annular structure is also nested within the regular pentagonal annular structure. At least when the ultrasound device is working, micro vortices generated in the liquid within the synthesis channel, distributed at the positions of the inner and outer sides of the regular pentagon, at the positions of the nested second regular pentagon, or at the positions of the inner and outer sides of the second regular pentagon, mix the two liquids within the synthesis channel to generate the target liposomes.

2. A liposome synthesis apparatus for implementing the liposome synthesis method of claim 1, characterized in that, include: The Y-shaped flow channel includes two injection channels and a synthesis channel connected to the confluence of the two injection channels. The two injection channels are used to inject two liquids, an organic phase and an aqueous phase. A regular pentagonal ultrasonic device is disposed at the bottom of the synthesis channel and at the confluence of the two sample inlet channels and the synthesis channel, wherein two adjacent sides of the regular pentagonal ultrasonic device face the two sample inlet channels respectively; the shape of the ultrasonic device forms an internally hollowed-out regular pentagonal annular structure, one outer vertex of which is located at the boundary line of the two liquids flowing into the synthesis channel; a second regular pentagon or a second regular pentagonal annular structure is also nested inside the regular pentagonal annular structure; At least when the ultrasound device is working, micro vortices generated in the liquid within the synthesis channel, distributed at the positions of the inner and outer sides of the regular pentagon, at the positions of the nested second regular pentagon, or at the positions of the inner and outer sides of the second regular pentagon, mix the two liquids within the synthesis channel to generate the target liposomes.

3. The apparatus according to claim 2, characterized in that, The synthesis channel is linear in shape, with a width of 100-160 micrometers and a length of 500-6 millimeters.

4. The apparatus according to claim 3, characterized in that, The ultrasonic device adopts a continuous working mode or a pulse working mode, with a power of 250 milliwatts to 400 milliwatts; The flow rate ratio of the organic phase to the aqueous phase is 1:1 to 1:9.

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