KR102492420B1 - Lipid nanoparticles manufacturing Chip, Lipid nanoparticles manufacturing System having the same, and Lipid nanoparticles manufacturing method - Google Patents

Abstract

The chip for producing lipid nanoparticles is connected to a mixer unit that mixes a first raw material containing mRNA and a second raw material containing lipid to form a mixed solution, and the mixer unit, and dilutes the mixed solution using a diluent. It includes a dilution unit and a concentration unit connected to the dilution unit and concentrating Lipid Nanoparticles (LNP) from the diluted mixed solution to obtain a concentrated solution.

Description

Lipid nanoparticle manufacturing chip, lipid nanoparticle manufacturing system including the same, and lipid nanoparticle manufacturing method {Lipid nanoparticles manufacturing Chip, Lipid nanoparticles manufacturing System having the same, and Lipid nanoparticles manufacturing method}

The present invention relates to a chip for preparing lipid nanoparticles, a system for preparing lipid nanoparticles including the chip for preparing lipid nanoparticles, and a method for preparing lipid nanoparticles, and more particularly, to manufacturing lipid nanoparticles containing active ingredients such as mRNA. It relates to a lipid nanoparticle preparation chip, a lipid nanoparticle preparation system including the lipid nanoparticle preparation chip, and a lipid nanoparticle preparation method including the active ingredient.

mRNA (messenger RNA) is a material at the stage before protein synthesis, and contains genetic information. Since mRNA is accessible to various therapeutic agents, it can be used as a preventive or therapeutic vaccine, and proteins that are lacking can also be synthesized through mRNA. The advantage of mRNA therapeutics is that they do not have to be delivered to the nucleus compared to DNA, and they are not inserted into the genome, so they do not cause permanent genetic diseases, so they are highly safe. In addition, it is possible to synthesize mRNA into proteins that are lacking inside cells that are inaccessible to protein therapeutics. mRNA can have various sizes depending on the protein it expresses, and exists as a single strand. mRNA made from DNA escapes from the nucleus into the cytoplasm and meets with livesomes to produce proteins. Although mRNA is in the limelight as a next-generation gene therapy, since it is single-stranded, its stability is very low, so it is rapidly degraded by nucleases in the blood and rapidly excreted from the body through the kidneys. It is known.

In the treatment using anionic drugs including nucleic acids, safe and efficient drug delivery technologies have been studied for a long time, and various delivery systems and delivery technologies have been developed. A variety of studies related to mRNA vaccines using a method of encapsulating and delivering mRNA in lipid nanoparticles have been conducted, and research on mass production systems is ongoing.

Recently, in fields such as pharmaceuticals, vaccines, and DDS (drug delivery systems), a method of encapsulating and delivering mRNA in lipid nanoparticles has been in the limelight, but there are difficulties in mass production. There is a bulk production method in which turbulence is generated in a large vessel to mix, dilute, and concentrate, but it is difficult to obtain uniform quality of the prepared lipid nanoparticles and it is difficult to control the manufacturing process.

Accordingly, the technical problem of the present invention is focused on this point, and an object of the present invention is to provide a chip for preparing lipid nanoparticles for preparing lipid nanoparticles.

Another object of the present invention is to provide a lipid nanoparticle manufacturing system including the lipid nanoparticle manufacturing chip.

Another object of the present invention is to provide a method for preparing lipid nanoparticles.

A chip for producing lipid nanoparticles according to an embodiment for realizing the object of the present invention described above is a mixer unit for forming a mixed solution by mixing a first raw material containing an active ingredient and a second raw material containing a lipid, A dilution unit connected to the mixer unit and diluting the mixed solution using a diluent, and a concentrating unit connected to the dilution unit and concentrating Lipid Nanoparticles (LNP) from the diluted mixed solution to obtain a concentrated solution. do.

In one embodiment of the present invention, the enrichment unit includes a main flow path connected to the dilution unit, an ion exchange channel in contact with the main flow path, a buffer solution channel spaced apart from the main flow path and in contact with the ion exchange channel, and the main flow path. It may include a lipid nanoparticle obtaining flow path connected to and obtaining the concentrate, and a recovery flow path connected to the main flow path and recovering solutions other than the concentrate.

In one embodiment of the present invention, the ion exchange channel of the enrichment unit may have a V-shape disposed above or below the main flow path and having an appendix disposed in a flow direction on a plane.

In one embodiment of the present invention, the lipid nanoparticle collection passage and the recovery passage have a smaller cross-sectional area than the main passage and may be disposed parallel to each other.

In one embodiment of the present invention, the mixer unit is connected to a first raw material supply passage through which the first raw material is supplied, a second raw material supply passage through which the second raw material is supplied, and the first and second raw material supply passages. It may include a mixed flow path that becomes.

In one embodiment of the present invention, the dilution unit includes a first flow path connected to the mixing flow path of the mixer unit, a dilution flow path providing the dilution solution, and a dilution space connected to the dilution flow path and the first flow path. can do.

In one embodiment of the present invention, the active ingredient may be a nucleic acid.

In one embodiment of the present invention, the active ingredient may be any one of mRNA, miRNA, siRNA, DNA and CRISPR.

In one embodiment of the present invention, the first raw material may include mRNA and water, the second raw material may include lipid and ethanol, and the diluent may include deionized water. .

A lipid nanoparticle manufacturing system according to an embodiment for realizing the object of the present invention described above is a first raw material supply unit for supplying a first raw material, a second raw material supply unit for supplying a second raw material, the first raw material and the first raw material supply unit. 2. A chip for preparing lipid nanoparticles including a mixer unit for mixing raw materials, a dilution unit for diluting the mixed solution, and a concentration unit for concentrating lipid nanoparticles from the diluted mixture to obtain a concentrated solution, providing a dilution solution to the chip for preparing lipid nanoparticles. and a recovery unit for recovering a solution other than the concentrate from the chip for producing the lipid nanoparticles.

In one embodiment of the present invention, the enrichment unit of the chip for producing lipid nanoparticles includes a main flow path connected to the dilution unit, an ion exchange channel in contact with the main flow path, and spaced apart from the main flow path and in contact with the ion exchange channel. It may include a buffer solution channel, a lipid nanoparticle collection channel connected to the main channel to obtain the concentrate, and a recovery channel connected to the main channel to recover solutions other than the concentrate.

In one embodiment of the present invention, the ion exchange channel of the enrichment part of the chip for producing lipid nanoparticles is disposed above or below the main flow path and has a v-shape so that the tip is disposed in the direction of flow of the fluid on a plane. can

In one embodiment of the present invention, the dilution unit of the chip for producing lipid nanoparticles includes a first flow path connected to the mixer unit, a dilution flow path providing the dilution solution, and a dilution flow path connected to the first flow path. Dilution space may be included.

A lipid nanoparticle manufacturing method according to an embodiment for realizing the object of the present invention described above includes preparing a first raw material containing an active ingredient and a second raw material containing a lipid, the first raw material and It may include mixing the second raw material to form lipid nanoparticles containing mRNA, and a post-processing step of manufacturing a final product by filtering and filling a solution containing the lipid nanoparticles into individual containers. The step of forming the lipid nanoparticles is performed on a chip for producing lipid nanoparticles having a flow path, and the mixing step of mixing the first raw material and the second raw material to form a mixed solution, the mixed solution using a diluent It may include a dilution step of diluting, and a concentration step of obtaining a concentrated solution by concentrating the lipid nanoparticles from the diluted mixed solution.

In one embodiment of the present invention, the concentration step of the formation of lipid nanoparticles is performed by applying a voltage to a buffer solution channel connected to an ion exchange channel in contact with the flow path of the chip for preparing lipid nanoparticles, so that the solution in the flow path is By forming an ion-rich zone and an ion-depleted zone, the lipid nanoparticles containing the active ingredient can be concentrated in a specific region within the flow path.

In one embodiment of the present invention, the chip for producing lipid nanoparticles is connected to a mixer unit and the mixer unit for mixing the first raw material and the second raw material to form the mixed solution, and the mixed solution using the diluent. It may include a dilution unit for diluting, and a concentration unit connected to the dilution unit and concentrating the lipid nanoparticles from the diluted mixed solution.

In one embodiment of the present invention, the enrichment unit of the chip for producing lipid nanoparticles includes a main flow path connected to the dilution unit, an ion exchange channel in contact with the main flow path, and spaced apart from the main flow path and in contact with the ion exchange channel. It may include a buffer solution channel, a lipid nanoparticle collection channel connected to the main channel to obtain the concentrate, and a recovery channel connected to the main channel to recover solutions other than the concentrate.

According to the embodiments of the present invention, since mixing, dilution, and concentration for lipid nanoparticle production are all performed on a chip for lipid nanoparticle production, it is easy to control the size and uniformity of lipid nanoparticles, thereby producing high-quality lipid nanoparticles can do.

However, the effects of the present invention are not limited to the above effects, and may be expanded in various ways without departing from the spirit and scope of the present invention.

1 is a schematic diagram of a lipid nanoparticle production system according to an embodiment of the present invention.
FIG. 2 is a view showing various examples of a mixer unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .
FIG. 3 is a view showing various examples of a dilution unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .
FIG. 4 is a view for explaining the enrichment part of the chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .
5 is a view showing various examples of an enrichment unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .
6 is a view showing an embodiment of an enrichment unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .
7 is a view for explaining the fluid flow and concentrating effect according to the shape of the divorce exchange part of the concentrating part of the chip for producing lipid nanoparticles according to an embodiment of the present invention.
8 is a flowchart showing a method for preparing lipid nanoparticles according to an embodiment of the present invention.
9 is a flowchart showing in detail the steps of preparing LNPs on a chip in the method for preparing lipid nanoparticles of FIG. 8 .

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to the drawings.

Since the present invention may have various changes and various forms, specific embodiments are illustrated in the drawings and described in detail in the text. However, it should be understood that this is not intended to limit the present invention to the specific disclosed form, and includes all modifications, equivalents, and substitutes included in the spirit and scope of the present invention.

1 is a schematic diagram of a lipid nanoparticle production system according to an embodiment of the present invention. FIG. 2 is a view showing various examples of a mixer unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 . FIG. 3 is a view showing various examples of a dilution unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 . FIG. 4 is a view for explaining the enrichment part of the chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .

1 to 4, the lipid nanoparticle manufacturing system includes a first raw material providing unit 10 including mRNA, a second raw material providing unit 20 including lipid (Lipid), a dilution solution providing unit 30, It includes a lipid nanoparticle acquisition unit 40, a first recovery unit 50, a second recovery unit 60, and a lipid nanoparticle manufacturing chip 100.

The first raw material supply unit 10 stores the first raw material and provides the first raw material to the chip 100 for producing lipid nanoparticles. The first raw material may be a solution containing mRNA. For example, the first raw material may include mRNA and water.

The second raw material supply unit 20 stores the second raw material and provides the second raw material to the chip 100 for producing lipid nanoparticles. The second raw material may be a solution containing lipid. For example, the second raw material may include lipids and ethanol.

The lipid nanoparticle preparation chip 100 may mix, dilute, and concentrate the first raw material and the second raw material to prepare a lipid nanoparticle concentrate containing mRNA. The chip 100 for preparing lipid nanoparticles includes a mixer unit 110, a dilution unit 120, and a concentration unit 130.

The mixer unit 110 may mix the first raw material and the second raw material to prepare a mixed pressure solution. As the mixer unit 110, a microfluidic mixer commonly used in a microchannel, such as a chaotic mixer or a herringbone mixer, may be used. At this time, a liquid mixture containing lipid nanoparticles can be prepared by self-aligning of lipid and mRNA at the interface of the two fluids through mixing of the first raw material and the second raw material in the flow path.

For example, the mixer unit 110 includes a first raw material supply channel 112 through which the first raw material is supplied, a second raw material supply channel 114 through which the second raw material is supplied, A mixing passage 116 connected to the first and second raw material supply passages may be included. The first raw material supply passage 112 and the second raw material supply passage 114 meet each other at the merging point JP so that the first raw material and the second raw material may be mixed.

The dilution unit 120 may dilute the mixed solution. The dilution unit 120 includes a first flow path 123 connected to the mixing flow path 116 of the mixer unit 110, a dilution flow path 122 providing the dilution solution, and the dilution flow path 122 and the dilution flow path 122. A dilution space 124 connected to the first flow path 123 may be included. The dilution space 124 may have various shapes, such as a line shape (see FIG. 3(a)), a serpentine chamber shape (see FIG. 3(b)), and the like. The dilution solution providing unit 30 may provide the dilution solution through the dilution passage 122 of the dilution unit 120 of the chip 100 for preparing lipid nanoparticles. The diluent may include deionized water.

Since the solution mixed in the mixing unit 110 contains the solvent of the second raw material, for example, ethanol at a high concentration, dilution to a desired concentration is required. The concentration of the ethanol may be lowered to a desired level while passing through the dilution unit 120 .

Although not shown, the dilution unit 120 may be implemented in the flow path structure of the mixing unit 110 . That is, a structure in which the first raw material, the second raw material, and the diluting liquid are input may be possible by configuring the mixing and diluting unit so that the mixer unit and the dilution unit are implemented simultaneously rather than sequentially.

The concentration unit 130 may concentrate the lipid nanoparticles from the mixed solution diluted in the dilution unit 120 to obtain a concentrated solution. The enrichment unit 130 is spaced apart from the main flow path 132 connected to the dilution unit 120, an ion exchange channel (IM) in contact with the main flow path 132, and the main flow path 132, and the ion exchange Buffer solution channels 134a and 134b in contact with the channel IM, connected to the main flow path 132, and a lipid nanoparticle obtaining flow path 136 for obtaining the concentrate, and connected to the main flow path 132, Recovery channels 138a and 138b for recovering solutions other than the concentrate may be included. The ion exchange channel (IM) may be a charged nanochannel for exchanging cations or anions made of a nanoporous material.

In the enrichment unit 130, lipid nanoparticles having a size of 100 nm or less may be separated and concentrated using an ion concentration polarization (ICP) phenomenon.

In the ICP phenomenon, when channels are located on both sides with an ion exchange membrane in the middle and a voltage is applied to both ends of the channel, in the case of a cation exchange channel, cations move to the ion exchange membrane to form an ion enrichment zone, relatively opposite. It refers to a natural phenomenon in which an ion depletion zone (IDZ) is formed in The ion depletion zone generates a strong repulsive force to push ions/molecules regardless of polarity, and lipid nanoparticles can be separated and concentrated using this.

According to the present embodiment, buffer solution channels 134a and 134b, through which the buffer solution flows, are formed on both sides of the second direction D2 of the main flow path 132 through which the fluid flows along the first direction D1, and both sides When an appropriate voltage is applied to the buffer solution channels 134a and 134b of , ions move through the ion exchange channel IM, and an ion depletion zone (see IDZ in FIG. 8) is formed along the second direction D2. Formed on both sides of the main flow path 132, lipid nanoparticles having an electro-dense core are concentrated in the center of the main flow path 132, and pass through the lipid nanoparticle acquisition flow path 136. That is, the cation component is used as a driving source of the ion concentration polarization phenomenon and is discharged to the outside of the chip through the main channel 132 and the ion exchange channel IM through the buffer solution channels 134a and 134b. The remaining positive cation component is affected by the ion depletion region and the electric field formed at the interface of the ion depletion region, and the path is set according to the properties of the particle to the lipid nanoparticle acquisition channel 136 and the recovery channel 138a, 138b and can be moved selectively.

Accordingly, the concentration of lipid nanoparticles increases in the fluid passing through the lipid nanoparticle collection channel 136, and fluids other than the lipid nanoparticles can be recovered in the recovery channels 138a and 138b.

According to an embodiment of the present invention, mixing to prepare lipid nanoparticles, dilution to dilute a solvent such as ethanol, and concentration to increase lipid nanoparticle concentration can all be performed on the chip 100 for preparing lipid nanoparticles.

The ion exchange channel (IM) may have various shapes (see FIGS. 5 and 6 ), and for example, the ion exchange channel (IM) is disposed above or below the main flow path 132 and is I on a plane. It may have a V-shape so that the attachment is disposed in the direction of flow of the ruler or fluid.

A sum of widths of the recovery passages 138a and 138b in the second direction D2 may be smaller than the width of the main passage 132 . For example, the recovery passages 138a and 138b may have an I-shaped channel structure as a whole since the main passage 132 is divided into three passages and formed in parallel. The area occupied by the enrichment unit 130 is reduced, space integration efficiency is improved compared to a structure in which channels are radially formed, fluid flow is improved, and operation is possible with low power, thereby improving production efficiency. In addition, the same or superior efficiency can be obtained with lower power consumption compared to the conventional structure in which the ion depletion zone is generated on both sides and the ion depletion zone is formed only on one side of the flow path, and it can be used as an on/off valve by adjusting the power. . (See (b) of FIG. 8)

Meanwhile, in this embodiment, a structure in which the main flow path is divided into three branches, including one lipid nanoparticle obtaining flow path and two recovery flow paths, is described, but is not limited thereto. For example, the main passage may be divided into 5 or 7 branches, and the size and width of each of the divided passages may be different.

Here, the chip 100 for preparing the lipid nanoparticles may be manufactured by various known methods. For example, it may be formed by stacking a plurality of substrates having grooves corresponding to passages. For example, an ion exchange channel may be formed on a lower substrate, and an upper substrate having a groove formed on a lower surface may be formed by bonding to the lower substrate, but is not limited thereto. As for the material of the substrate, various substrates capable of forming a flow path pattern, such as a glass substrate, a plastic substrate, a silicon substrate, etc. Methods may be used, and various known methods such as a laser bonding method, a thermal compression method, an adhesion method, and a lamination method may be used for bonding of a plurality of substrates.

According to the present invention, since mixing, dilution, and concentration for lipid nanoparticle production are all performed on a chip for lipid nanoparticle production (on chip process), the fluid is controlled in a laminar flow state, and thus lipid nanoparticles It is easy to control the size and uniformity of the lipid nanoparticles of high quality. According to the bulk size manufacturing method in which turbulence cannot be controlled, it is difficult to control the size and uniformity of lipid nanoparticles because fine fluid control is impossible. In the conventional lipid nanoparticle manufacturing method using the chip method, only the mixing of mRNA and lipid is performed on the chip, and the steps for diluting organic solvents such as ethanol and concentrating the lipid nanoparticles require separate processes. Since it proceeds in the bulk size through the process, it is not easy to control the lipid nanoparticle manufacturing process compared to the present invention.

In particular, in the case of a solution containing lipid nanoparticles in the initial stage manufactured on a microfluidics chip, the flow rate conditions in the manufacturing stage, in particular, the ratio of the organic phase in which lipids are dissolved in ethanol to the aqueous phase (Flow Rate Ratio) ), a certain concentration of ethanol is necessarily included. Since the concentration of ethanol contained is an unstable state that can affect the lipid of nanoparticles in the early stages, effective post-process application has a great effect on the production of stable lipid nanoparticles. Through the bulk size process, lipid nanoparticles at an early stage are discharged to the outside of the chip and may not maintain stability until diluted in bulk size through a tubing line. However, according to embodiments of the present invention, lipid nanoparticles Since mixing, dilution, and concentration of nanoparticles are all performed on a chip (on chip process), this problem can be solved.

5 is a view showing various examples of an enrichment unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .

(a) of FIG. 5 shows a case in which the ion exchange channel IM has an I-shape extending in a second direction D2 perpendicular to the first direction D1 in which the main flow path 132 extends. has been When viewed from the side, the ion exchange channel IM has a predetermined height on the lower side of the main flow path 132 in a third direction D3 perpendicular to the first and second directions D1 and D2. can be formed to have

Meanwhile, other configurations of the enrichment unit are omitted since they are substantially the same as those of the enrichment unit described above.

Figure 5 (b) is an embodiment in which the ion exchange channel (IM) is formed in a V shape, Figure 5 (c) is an embodiment in which two V-shaped ion exchange channels (IM1, IM2) are formed, Figure 5 (d) is an embodiment in which an I-shaped ion exchange channel (IM1) and a V-shaped ion exchange channel (IM2) are sequentially formed, and (e) of FIG. 5 is a V-shaped ion exchange channel (IM) These are drawings showing an embodiment in which the plurality is formed.

6 is a view showing an embodiment of an enrichment unit of a chip for preparing lipid nanoparticles of the lipid nanoparticle manufacturing system of FIG. 1 .

Referring to FIG. 6, when viewed from the side, the ion exchange channels IM1 and IM2 are formed on the upper and lower surfaces of the main flow path 132, and the ion depletion zone can be formed more efficiently.

7 is a view for explaining the fluid flow and concentrating effect according to the shape of the divorce exchange part of the concentrating part of the chip for producing lipid nanoparticles according to an embodiment of the present invention.

Referring to FIG. 7 , it can be confirmed that the concentration effect is maximized and power consumption is reduced when the ion exchange membrane has a V-shape rather than an I-shape.

FIG. 7 (a) is a diagram showing the fluid flow and concentrating effect of the I-shaped ion exchange channel, and FIG. 7 (b) is a diagram showing the fluid flow and concentrating effect of the V-shaped ion exchange channel.

Figure 7 (c) is the result of the power consumption comparison experiment according to the channel shape, Figure 7 (d) is an experiment confirming that the concentration of lipid nanoparticles can be controlled (valve effect) according to the applied voltage / current is the drawing shown. 7(e) is a view showing an experiment confirming that the particle separation direction can be changed according to the formation of an electric field.

According to (c) to (e) of FIG. 7 , the enrichment unit of the chip for preparing seismic nanoparticles according to an embodiment of the present invention not only has a function of concentrating the lipid nanoparticles, but also has a form of an on/off valve to turn on/off the obtained lipid nanoparticles. It can be controlled as, and if necessary, it is possible to obtain the required particles not only through the lipid nanoparticle obtaining channel but also through the recovery channel.

8 is a flowchart showing a method for preparing lipid nanoparticles according to an embodiment of the present invention. 9 is a flowchart showing in detail the step of preparing lipid nanoparticles on a chip of the method for preparing lipid nanoparticles of FIG. 8 .

8 and 9, the method for preparing lipid nanoparticles may include preparing raw materials (S100), preparing lipid nanoparticles on a chip (S200), and post-processing (S300).

In the step of preparing the raw material (S100), a first raw material including mRNA and a second raw material including lipid may be prepared.

In the step of preparing lipid nanoparticles on the chip (S200), lipid nanoparticles containing mRNA may be formed by mixing the first raw material and the second raw material.

In the post-processing step (S300), a final product may be prepared by diafiltration of the solution containing the lipid nanoparticles, additional concentration to a required concentration, and filling into individual containers.

At this time, the step of forming the lipid nanoparticles (S100) is performed on a chip for preparing lipid nanoparticles on which a channel is formed, and may include a mixing step (S110), a dilution step (S120), and a concentration step (S130). there is. As the chip for preparing the lipid nanoparticles, the chip for lipid nanoparticles described in FIG. 1 may be used. For example, the chip for preparing the lipid nanoparticles may include a mixer unit, a dilution unit, and a concentration unit.

In the mixing step (S110), a mixed solution may be formed by mixing the first raw material and the second raw material. The mixing step (S110) may be performed in the mixer unit of the chip for preparing the lipid nanoparticles.

In the dilution step (S120), the mixed solution may be diluted using a dilution solution. The dilution step (S120) may be performed in the dilution unit of the chip for preparing the lipid nanoparticles.

In the concentration step (S130), a concentrate may be obtained by concentrating the lipid nanoparticles from the diluted mixed solution. The enrichment step (S130) may be performed in the enrichment unit of the chip for preparing the lipid nanoparticles. Specifically, the concentrated solution applies a voltage to a buffer solution channel connected to an ion exchange channel in contact with the flow path of the chip for producing lipid nanoparticles to form an ion-rich zone and an ion-depleted zone in the solution within the flow path, including mRNA The lipid nanoparticles may be concentrated in a specific region within the flow path.

Although described with reference to the above embodiments, those skilled in the art will understand that the present invention can be variously modified and changed without departing from the spirit and scope of the present invention described in the claims below. You will be able to.

10: first raw material supply unit 20: second raw material supply unit
30: diluent supply unit 40: lipid nanoparticle acquisition unit
50, 60: first and second recovery units 100: chip for producing lipid nanoparticles

Claims (17)

A mixer unit for mixing a first raw material containing an active ingredient and a second raw material containing a lipid to form a mixed solution;
a dilution unit connected to the mixer unit and diluting the mixed solution using a dilution liquid; and
A concentration unit connected to the dilution unit and concentrating lipid nanoparticles (LNP) from the diluted mixture to obtain a concentrated solution,
The mixer unit, the dilution unit and the enrichment unit are formed on one substrate,
The enrichment unit includes a main flow path connected to the dilution unit, a lipid nanoparticle obtaining flow path connected to the main flow path and obtaining the concentrate, and a recovery flow path connected to the main flow path and recovering solutions other than the concentrate, A chip for producing lipid nanoparticles, characterized in that the concentrated solution and the solution other than the concentrated solution are continuously obtained and recovered at the same time. According to claim 1,
The enrichment part
an ion exchange channel in contact with the main flow path; and
A chip for producing lipid nanoparticles, characterized in that it further comprises a buffer solution channel spaced apart from the main flow path and in contact with the ion exchange channel. According to claim 2,
The ion exchange channel of the enrichment part is disposed above or below the main flow path and has a v-shape so that the attachment is disposed in the direction of flow of the fluid on a plane. According to claim 2,
The lipid nanoparticle preparation chip, characterized in that the lipid nanoparticle obtaining flow path and the recovery flow path have a smaller cross-sectional area than the main flow path and are disposed parallel to each other. According to claim 1,
the mixer part
A first raw material supply passage through which the first raw material is supplied, a second raw material supply passage through which the second raw material is supplied, and a mixing passage connected to the first and second raw material supply passages. Chips for making particles. According to claim 5,
The dilution part,
a first flow path connected to the mixing flow path of the mixer unit;
a dilution passage for providing the dilution solution; and
A chip for producing lipid nanoparticles, comprising a dilution space connected to the dilution passage and the first passage. According to claim 1,
The active ingredient is a chip for producing lipid nanoparticles, characterized in that nucleic acid. According to claim 1,
The active ingredient is a chip for producing lipid nanoparticles, characterized in that any one of mRNA, miRNA, siRNA, DNA and CRISPR. According to claim 1,
The first raw material includes mRNA and water, the second raw material includes lipid and ethanol, and the diluent includes deionized water. A first raw material supply unit supplying a first raw material;
a second raw material supply unit supplying a second raw material;
A chip for producing lipid nanoparticles comprising a mixer unit for mixing the first raw material and the second raw material to form a mixed solution, a dilution unit for diluting the mixed solution, and a concentration unit for concentrating lipid nanoparticles from the diluted mixture to obtain a concentrated solution. ;
a dilution solution providing unit providing a dilution solution to the chip for preparing the lipid nanoparticles; and
A recovery unit for recovering solutions other than the concentrate from the chip for producing lipid nanoparticles,
The mixer part, the dilution part and the enrichment part of the chip for producing lipid nanoparticles are formed on one substrate,
The enrichment unit includes a main flow path connected to the dilution unit, a lipid nanoparticle obtaining flow path connected to the main flow path and obtaining the concentrate, and a recovery flow path connected to the main flow path and recovering solutions other than the concentrate, The lipid nanoparticle production system, characterized in that the concentrated solution and the solution other than the concentrated solution are continuously obtained and recovered at the same time. According to claim 10,
The enrichment part of the chip for preparing the lipid nanoparticles
an ion exchange channel in contact with the main flow path; and
A lipid nanoparticle manufacturing system further comprising a buffer solution channel spaced apart from the main flow path and in contact with the ion exchange channel. According to claim 11,
The lipid nanoparticle manufacturing system, characterized in that the ion exchange channel of the enrichment part of the chip for producing lipid nanoparticles is disposed above or below the main flow path and has a v-shape so that the attachment is disposed in the direction of flow of the fluid on a plane. . According to claim 10,
The dilution part of the chip for preparing the lipid nanoparticles,
a first flow path connected to the mixer unit;
a dilution passage for providing the dilution solution; and
A lipid nanoparticle manufacturing system comprising a dilution space connected to the dilution passage and the first passage. Preparing a first raw material containing an active ingredient and a second raw material containing a lipid;
Forming lipid nanoparticles containing the active ingredient by mixing the first raw material and the second raw material; and
A post-processing step of preparing a final product by filtering and filling the solution containing the lipid nanoparticles into individual containers,
The step of forming the lipid nanoparticles is performed on a chip for preparing lipid nanoparticles in which a flow path is formed,
a mixing step of mixing the first raw material and the second raw material to form a liquid mixture;
A dilution step of diluting the mixed solution using a dilution solution; and
A concentration step of concentrating the lipid nanoparticles from the diluted mixture to obtain a concentrated solution, wherein the mixing step, the dilution step, and the concentration step are performed on one substrate, which is a component of the chip for preparing the lipid nanoparticles,
The chip for producing the lipid nanoparticles
a mixer unit mixing the first raw material and the second raw material to form the mixed solution;
a dilution unit connected to the mixer unit and diluting the mixed solution using the dilution liquid; and
A concentration unit connected to the dilution unit and concentrating the lipid nanoparticles from the diluted mixture;
The enrichment unit includes a main flow path connected to the dilution unit, a lipid nanoparticle obtaining flow path connected to the main flow path, and a recovery flow path connected to the main flow path,
In the concentration step,
Obtaining the concentrate through the lipid nanoparticle obtaining flow path, recovering a solution other than the concentrate through the recovery flow path, lipid nanoparticles characterized in that the collection of the concentrate and the recovery of the solution other than the concentrate are simultaneously and continuously manufacturing method. According to claim 14,
The concentration step of the lipid nanoparticle formation step
A voltage is applied to a buffer solution channel connected to an ion exchange channel in contact with the main channel of the chip for producing lipid nanoparticles to form an ion-rich zone and an ion-depleted zone in the solution in the main channel, A method for producing lipid nanoparticles, characterized in that the lipid nanoparticles are concentrated in a specific region within the flow path. According to claim 14,
The enrichment part of the chip for preparing the lipid nanoparticles
an ion exchange channel in contact with the main flow path;
A lipid nanoparticle manufacturing method further comprising a buffer solution channel spaced apart from the main flow path and in contact with the ion exchange channel. According to claim 16,
The concentration step
Applying a voltage to the buffer solution channel to form an ion-rich zone and an ion-depleted zone in the solution in the main channel to concentrate the lipid nanoparticles containing the active ingredient in a specific region in the main channel A method for preparing lipid nanoparticles.

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