WO2025071365A1 - Flow reactor and biological polymer synthesis system including same - Google Patents

Abstract

The present invention relates to a flow reactor and a biological polymer synthesis system including same, the system comprising: a mixing reservoir in which at least one from among an amino acid, a reagent, and an additive is mixed; a reactor into which a fluid discharged from the mixing reservoir flows; a temperature regulator disposed between the mixing reservoir and the reactor so as to regulate the temperature of the fluid flowing into the reactor; and a pump for providing driving force such that the fluid flows from the mixing reservoir to the reactor, wherein the reactor includes: a column having an inlet in through which the fluid flows and an outlet through which the fluid is discharged; and a composite solid support disposed in the column.

Description

Flow reactor and biological polymer synthesis system comprising the same

The present invention relates to a flow reactor and a biological polymer synthesis system.

Peptide drugs generate billions of dollars in sales annually in the fields of diabetes, obesity, and oncology, and are expanding into the development of new drugs for emerging diseases such as cardiovascular and neurodegenerative diseases. Currently, the shortcoming of peptides, which is a shortcoming of short half-life in the body, has been solved by methods such as the introduction of unnatural amino acids. Various peptide drugs with a dosing cycle of more than one week are already on the market, and many candidates for oral administration are undergoing clinical trials with the U.S. Food and Drug Administration. Accordingly, the production demand for peptide drugs is increasing by nearly 10% every year, and there is an urgent need for a new synthesis platform to respond to increased productivity and ESG issues such as regulations on the use of DMF used in traditional chemical synthesis methods.

Peptide drugs are largely manufactured using biological or chemical methods. Biological methods using genetically modified plants or animals have the disadvantage that it is difficult to produce peptide drugs with extended half-lives using non-natural amino acids or PEGylation, which are recent trends, and that it is difficult to remove various endogenous contaminants derived from organisms or exogenous contaminants generated during the process.

In the case of liquid phase reactions in chemical methods, the process cost is low, but only peptides composed of 10 or fewer amino acids can be synthesized, so it is usually impossible to produce peptide drugs composed of 20 or more amino acids. Solid Phase Peptide Synthesis (SPPS) is the most suitable method for producing PEGylated peptide drugs composed of 20-50 different types of natural/unnatural amino acids.

In order to increase the production capacity of various biological polymers including peptides, various types of reactors are being studied in the SPPS field. Representative reactors include batch type reactors and flow type reactors. For mass synthesis, batch type reactors have been gradually scaled up from 10-50 mL to 100-500 L units, and various solutions have been improved for transfer, stirring, pressure control, and filtering methods. In laboratory scale, flow reactors have recently been introduced, and it has been shown that rapid peptide synthesis with high purity and yield is possible.

However, batch reactors have problems such as differences in temperature transfer and time until uniform mixing between the center and surface of the reaction solution because the reactor size varies depending on the reaction scale, and low reproducibility of reaction temperature/mixing, making it difficult to control yield and quality due to reproducibility.

On the other hand, a general flow reactor has the advantage of excellent reaction reproducibility as it passes through a tubular reactor and is mixed at high speed to increase the reaction surface area, thereby inducing uniform temperature transfer and chemical reaction, and easy quality control and yield control due to high reaction reproducibility.

The currently developed flow reactor has the advantage of rapid synthetic reaction, but has the problem of a back pressure of 30-50 bar when using 200 mg PS/DVB. This high back pressure clearly shows that the synthetic scale cannot be significantly increased. In addition, the flow reactor developed by Vapourtek in the UK solved the high back pressure, which is a disadvantage of packed bead-type resin, by using an adjustable syringe, but has a very small usable flow rate, and it has limitations in that it is difficult to significantly increase the synthetic scale due to the nature of pressure control using a syringe.

In addition, existing particle-type resins have structural limitations in that they cannot support themselves due to the limitations of the particle structure, and they are moved and packed by the flow, which increases the back pressure within the flow reactor.

Accordingly, a new self-supporting solid support material suitable for a SPPS-type flow reactor and a biological polymer synthesis system using the same were developed.

The present invention aims to provide a SPPS type flow reactor having improved synthesis efficiency and yield and high durability and stability, and a biological polymer synthesis system including the same.

One embodiment of the present invention provides a biological polymer synthesis system, comprising: a mixing tank in which at least one of an amino acid, a reagent, and an additive is mixed; a reactor into which a fluid discharged from the mixing tank is introduced; a temperature controller disposed between the mixing tank and the reactor to control a temperature of the fluid introduced into the reactor; and a pump providing a driving force so that the fluid flows from the mixing tank to the reactor, wherein the reactor comprises a column having an inlet through which the fluid is introduced and an outlet through which the fluid is discharged; and a composite solid support disposed inside the column.

The flow reactor of the present invention and the biological polymer synthesis system including the same can provide a biological polymer synthesis process with improved efficiency and yield. Specifically, the biological polymer synthesis system has the characteristics of using a flow reactor loaded with a composite solid support, exhibiting a low pressure drop even at a high flow rate, and maintaining excellent efficiency and yield even in a variety of fluid temperature ranges.

In addition, the flow reactor of the present invention and the biological polymer synthesis system including the same have improved overall durability and stability of the equipment and can precisely control the process.

FIG. 1 is a diagram illustrating a biological polymer synthesis system according to one embodiment of the present invention.

Figure 2 is a drawing illustrating the reactor of Figure 1.

Figure 3 is a graph measuring pressure drop according to flow rate to compare the reactor of the present invention with a comparative example.

Figure 4 is a graph measuring the pressure drop according to the size of the reactor of the present invention.

Figures 5 and 6 are graphs measuring the pressure drop according to temperature in the reactor of the present invention and a comparative example.

Figure 7 shows the appearance of the core substrate before coating, and Figure 8 shows the appearance of the composite solid support.

Figures 9 to 11 show HPLC graphs for ACP synthesized according to flow rate using a composite solid support according to one specific example.

One embodiment of the present invention provides a biological polymer synthesis system, comprising: a mixing tank in which at least one of an amino acid, a reagent, and an additive is mixed; a reactor into which a fluid discharged from the mixing tank is introduced; a temperature controller disposed between the mixing tank and the reactor to control a temperature of the fluid introduced into the reactor; and a pump providing a driving force so that the fluid flows from the mixing tank to the reactor, wherein the reactor comprises a column having an inlet through which the fluid is introduced and an outlet through which the fluid is discharged; and a composite solid support disposed inside the column.

Additionally, the composite solid support may include a core substrate and a functional coating positioned on the core substrate.

Additionally, the composite solid support can be self-standing when the fluid passes through the column.

Additionally, the composite solid support may have a loading density of the functional coating of 0.01 mmol/g to 2 mmol/g.

In addition, when the fluid passes through the reactor, the pressure drop between the inlet and the outlet may be 10 bar or less at a flow rate of 40 C.V/min or less. Specifically, the pressure drop may be measured based on the pressure difference generated at the inlet and outlet of an empty column. In addition, the flow rate can be expressed in C.V (Column Volume) per minute, and the reactor has a flow rate of 40 C.V/min or less, specifically 40 C.V/min or less, 30 C.V/min or less, 20 C.V/min or less, 0.1 C.V/min to 40 C.V/min, 1 C.V/min to 40 C.V/min, 5 C.V/min to 40 C.V/min, 10 C.V/min to 40 C.V/min, 15 C.V/min to 40 C.V/min, 20 C.V/min to 40 C.V/min, 25 C.V/min to 40 C.V/min, 30 C.V/min to 40 C.V/min, 35 C.V/min to 40 C.V/min, 0.1 C.V/min to 30 C.V/min, 1 C.V/min to 30 C.V/min, 5 C.V/min to 30 C.V/min, 10 C.V/min to 30 C.V/min, 15 C.V/min to 30 C.V/min, 20 C.V/min to 30 C.V/min, 25 C.V/min to 30 C.V/min, 0.1 C.V/min to 20 C.V/min, 1 C.V/min to 20 C.V/min, 5 C.V/min to 20 C.V/min, 10 C.V/min to 20 C.V/min, 15 C.V/min to 20 C.V/min, 0.1 C.V/min to 10 C.V/min, 1 C.V/min to 10 C.V/min, or 5 C.V/min to At 10 C.V/min the pressure drop can be less than 10 bar. In addition, in the above flow rate range, the pressure drop is 10 bar or less, specifically, 10 bar or less, 5 bar or less, 3 bar or less, 1 bar or less, 0.5 bar or less, 0.1 bar or less, 0.05 bar or less, 0.01 bar to 10 bar, 0.05 bar to 10 bar, 0.1 bar to 10 bar, 0.5 bar to 10 bar, 1 bar to 10 bar, 3 bar to 10 bar, 5 bar to 10 bar, 0.01 bar to 5 bar, 0.05 bar to 5 bar, 0.1 bar to 5 bar, 0.5 bar to 5 bar, 1 bar to 5 bar, 3 bar to 5 bar, 0.01 bar to 3 bar, 0.05 bar to 3 bar, 0.1 bar to 3 bar, 0.5 bar to 3 bar, 1 bar to 3 bar, It can represent 0.01 bar to 1 bar, 0.05 bar to 1 bar, 0.1 bar to 1 bar, 0.5 bar to 1 bar, 0.01 bar to 0.5 bar, 0.05 bar to 0.5 bar, 0.1 bar to 0.5 bar, 0.01 bar to 0.1 bar or 0.05 bar to 0.1 bar.

In addition, the temperature of the fluid flowing into the reactor after passing through the temperature controller may be 120°C or lower. The biological polymer system of the present invention can synthesize a biological polymer with excellent purity and efficiency even in a variety of fluid temperature ranges, and the temperature of the fluid may be 120°C or lower, specifically, 120°C or lower, 100°C or lower, 80°C or lower, 60°C or lower, 25°C to 120°C, 50°C to 120°C, 70°C to 120°C, 100°C to 120°C, 25°C to 100°C, 50°C to 100°C, 70°C to 100°C, 25°C to 70°C, 50°C to 70°C, 25°C to 50°C, but is not limited thereto, and any temperature of the fluid used for biological polymer synthesis may be set without limitation.

Additionally, a valve may be further included between the reactor and the mixing tank to discharge waste.

In addition, the method may further include a main line connecting the mixing tank, the pump, the temperature controller and the reactor, and a first branch line branched from the main line between the mixing tank and the reactor and connected to the mixing tank.

Additionally, the fluid can have a circulating flow through the main line.

Additionally, it may further include a solvent storage tank disposed on the first branch line.

Additionally, the solvent storage tank can be connected to the valve and the mixing storage tank through the first branch line.

In addition, the system may further include a sensor disposed on the main line to sense at least one of temperature, pressure, and flow rate of the fluid flowing along the main line.

Additionally, the main line may further include a detector that detects the progress of synthesis of the biological polymer.

Additionally, a second branch line branching from the valve and discharging the waste may be further included.

The present invention can be modified in various ways and has various embodiments, and specific embodiments are illustrated in the drawings and described in detail in the detailed description. The effects and features of the present invention and the methods for achieving them will become clear with reference to the embodiments described in detail below together with the drawings. However, the present invention is not limited to the embodiments disclosed below, and can be implemented in various forms.

Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings. When describing with reference to the drawings, identical or corresponding components are given the same drawing reference numerals and redundant descriptions thereof are omitted.

In the examples below, the terms first, second, etc. are not used in a limiting sense but are used for the purpose of distinguishing one component from another.

In the examples below, singular expressions include plural expressions unless the context clearly indicates otherwise.

In the examples below, terms such as “include” or “have” mean that a feature or component described in the specification is present, and do not exclude in advance the possibility that one or more other features or components may be added.

In the following examples, when a part such as a film, region, component, etc. is said to be on or above another part, this includes not only the case where it is directly on top of the other part, but also the case where another film, region, component, etc. is interposed in between.

In the drawings, the sizes of components may be exaggerated or reduced for convenience of explanation. For example, the sizes and thicknesses of each component shown in the drawings are arbitrarily shown for convenience of explanation, and therefore the present invention is not necessarily limited to what is shown.

In the following embodiments, when it is said that a film, a region, a component, etc. are connected, it includes not only the cases where the films, regions, and components are directly connected, but also the cases where other films, regions, and components are interposed between the films, regions, and components and are indirectly connected. For example, when it is said in this specification that a film, a region, and a component, etc. are electrically connected, it includes not only the cases where the films, regions, and components, etc. are directly electrically connected, but also the cases where other films, regions, and components are interposed between them and are indirectly electrically connected. Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings.

The term “loading density” in this specification means “the number of moles of functional groups provided per unit mass of the composite solid support”, and may refer to a synthetic reaction site of a biological polymer according to the composition ratio of the polymer for biological polymer synthesis according to one embodiment.

In one embodiment, the composite solid support comprises a core substrate having a length of 1 mm or more and a shape of one or more dimensions; and a functional coating surrounding the core substrate, wherein a biological polymer may be synthesized in the functional coating.

In one embodiment, the functional coating may have a property of swelling in a solvent.

In one embodiment, the functional coating may include a functional group on the surface and inside. Specifically, the functional coating includes a functional group, and due to the swelling characteristic in a solvent, it is easy for a reactant to penetrate into the functional coating due to a concentration gradient. Accordingly, the functional groups on the surface and inside of the functional coating are easily exposed to the reactant, so that high synthesis efficiency can be exhibited even at a low loading density.

In one embodiment, the functional group may include at least one selected from an amine group, a carboxyl group, a hydroxyl group, a carbonyl group, an amino group, a thiol group, and a phosphoric acid group.

In one embodiment, the loading density of the reaction site of the functional coating may be 0.01 mmol/g to 2 mmol/g. Specifically, the loading density of the reaction site of the functional coating is 0.01 mmol/g to 2 mmol/g, 0.05 mmol/g to 2 mmol/g, 0.1 mmol/g to 2 mmol/g, 0.3 mmol/g to 2 mmol/g, 0.5 mmol/g to 2 mmol/g, 0.7 mmol/g to 2 mmol/g, 1 mmol/g to 2 mmol/g, 0.01 mmol/g to 1.7 mmol/g, 0.05 mmol/g to 1.7 mmol/g, 0.1 mmol/g to 1.7 mmol/g, 0.3 mmol/g to 1.7 mmol/g, 0.5 mmol/g to 1.7 mmol/g, 0.7 mmol/g to 1.7 mmol/g, 1.0 mmol/g to 1.7 mmol/g, 0.01 mmol/g to 1.5 mmol/g, 0.05 mmol/g to 1.5 mmol/g, 0.1 mmol/g to 1.5 mmol/g, 0.3 mmol/g to 1.5 mmol/g, 0.5 mmol/g to 1.5 mmol/g, 0.7 mmol/g to 1.5 mmol/g, 1.0 mmol/g to 1.5 mmol/g, 0.01 mmol/g to 1.2 mmol/g, 0.05 mmol/g to 1.2 mmol/g, 0.1 mmol/g to 1.2 mmol/g, 0.3 mmol/g to 1.2 mmol/g, 0.5 mmol/g to 1.2 mmol/g, 0.7 mmol/g to 1.2 mmol/g, 1.0 mmol/g to 1.2 mmol/g, 0.01 mmol/g to 1 mmol/g, 0.05 mmol/g to 1 mmol/g, It may be, but is not limited to, 0.1 mmol/g to 1 mmol/g, 0.3 mmol/g to 1 mmol/g, 0.5 mmol/g to 1 mmol/g or 0.7 mmol/g to 1 mmol/g.

In one embodiment, the functional coating may have an expansion rate per unit mass of 2 mL/g to 8 mL/g in water. Specifically, the functional coating may have an expansion rate per unit mass of 2 mL/g to 8 mL/g, 2 mL/g to 7 mL/g, 2 mL/g to 6 mL/g, 2 mL/g to 5 mL/g, 3 mL/g to 8 mL/g, 3 mL/g to 7 mL/g, 3 mL/g to 6 mL/g or 3 mL/g to 5 mL/g in water, but is not limited thereto. In addition, the functional coating causes swelling in a solvent other than water and may have different expansion rates per unit mass depending on the type of solvent.

In one embodiment, the core substrate may have solvent resistant, thermal resistant, or both properties. Unlike the functional coating, the core substrate does not have swelling properties and solvent-soluble properties, and thus can be self-standing even at high flow rates and can act as a support for a composite solid support. In addition, the core substrate has thermal resistance, and thus can act as a support without being changed even when the synthesis of a biological polymer is carried out at a high temperature.

In one embodiment, the component of the core substrate may include at least one selected from a polymer, a metal, and a ceramic. Specifically, the polymer may include at least one selected from, but is not limited to, high-density polyethylene (HDPE), low-density polyethylene (LDPE), linear low-density polyethylene (LLDPE), ultra-high molecular weight polyethylene (UHMWPE), polypropylene (PP), polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyethylene naphthalate (PEN), polyamide (PA), polyvinyl chloride (PVC), polyvinyl difluoride (PVDF), polyacetal, polycarbonate (PC), polyimide (PI), polyether ether ketone (PEEK), polyether sulfone (PES), polyphenylene oxide (PPO), and polyphenylene sulfide (PPS). The metal may include at least one selected from, but is not limited to, stainless steel, titanium, nickel, tantalum, zirconium, and alloys thereof. The ceramic may include at least one selected from, but is not limited to, fused silica, alumina, zirconia, silicon carbide, silicon nitride, boron nitride, and titanium diboride.

In one embodiment, the core substrate may have at least one shape selected from a one-dimensional shape, a two-dimensional shape, and a three-dimensional shape.

In one embodiment, the one-dimensional shape may include at least one selected from a staple fiber, a filament fiber, and a rod. As a specific example, the one-dimensional shape of the core substrate may include a fiber shape, in which case the core substrate is configured to have a length sufficient for biological polymer synthesis to occur, and the composite solid support is loaded into a reactor in an entangled form so as to be utilized in a biological polymer synthesis reaction.

In one embodiment, the two-dimensional shape may include at least one selected from a spunbond non-woven fabric, a meltblown non-woven fabric, a needle-punched non-woven fabric, a hydroentangled non-woven fabric, a woven fabric, a knitted fabric, a porous membrane, a polymeric film, and a mesh. As a specific example, the two-dimensional shape of the core substrate may include a form in which a plurality of the one-dimensionally shaped core substrates are randomly arranged. In addition, by forming pores between the plurality of the one-dimensionally shaped cores by the random arrangement, the two-dimensional shape may exhibit random porosity.

In one embodiment, the three-dimensional shape may include an open cell foam, a macropored sphere, or both.

In one embodiment, the length of one cross-section of the core substrate may be 1 mm or more, and may have a size sufficient for the functional coating to participate in the synthesis of the intended biological polymer. As a specific example, when the core substrate has a one-dimensional shape, it may be implemented so that a core substrate having a shape such as a fiber or the like can act as a support for a composite solid support through entanglement. In addition, when the core substrate has a two-dimensional or three-dimensional shape, it may act as a support in itself, such as a nonwoven fabric (two-dimensional) and a foam (three-dimensional). Therefore, the length of one cross-section of the core substrate may be 1 mm or more, and theoretically, a one-dimensional core substrate having an infinite length can also serve as a support.

In one embodiment, the functional coating may include a polymer of one or more main monomers and an active monomer. As a specific example, the functional coating may include a polymer of the first monomer and the active monomer, or a polymer of the first monomer, the second monomer, and the active monomer. In addition, the functional coating of the present invention may be a polymer formed through a polymerization reaction of one or more main monomers with an active monomer providing a reaction site as a main component.

In one embodiment, the first monomer may include at least one selected from a bisacrylamide-based crosslinking agent, a methacryloyl group, an alkenyl group-substituted triazine, and an acrylate-based crosslinking agent, and specifically, the first monomer may include a polyethylene glycol-based agent, for example, polyethylene glycol diacrylate. Specifically, the first monomer may be used without limitation as long as it is a material that can react or bind with an active monomer for exposing a functional group to the interior and/or surface in a functional coating. As a non-limiting example, the first monomer may include at least one selected from polyethylene glycol diacrylate, N,N'-methylenebisacrylamide (MBA), ethylene glycol dimethacrylate (EGDMA), poly(ethylene glycol) dimethacrylate (PEGDMA), glycidyl methacrylate (GMA), divinyl sulfone (DVS), triethylene glycol divinyl ether (TEGDVE), and diallyl phthalate (DAP). The second monomer may include at least one selected from 2-hydroxyethyl acrylate, It may include at least one selected from 2-hydroxyethyl methacrylate (HEMA), acrylic acid (AA), methyl methacrylate (MMA), ethyl acrylate (EA), butyl acrylate (BA), glycidyl methacrylate (GMA), and vinyl acetate (VAc).

In one embodiment, the active monomer may include at least one selected from among acrylate-based, acrylamide-based, and methacrylamide-based materials to provide a functional group to the interior and/or surface of the functional coating. As non-limiting examples, the active monomers include N-(2-aminopropyl) methacrylamide hydrochloride, Aminoethyl Methacrylate Hydrochloride (AEMA.HCl), 2-Aminoethyl Methacrylate (2-AEMA), N-(3-Aminopropyl) methacrylamide Hydrochloride (APMA), 4-Aminostyrene, N-(4-Aminophenyl)methacrylamide, N,N-Dimethylaminoethyl Methacrylate (DMAEMA), N,N-Dimethylaminopropyl Acrylamide (DMAPAA). and may include at least one selected from N-(2-aminoethyl)acrylamide hydrochloride.

In one embodiment, the composite solid support may include a homogeneous composite solid support, a heterogeneous composite solid support, or both. Specifically, in the homogeneous composite solid support, the functional coating may be formed as a polymer of the one or more main monomers and the active monomer. Additionally, in the heterogeneous composite solid support, the functional coating including the pulverized conventional solid support may be in a form coated on the core substrate. The above conventional solid support may include at least one selected from, but is not limited to, polystyrene/divinylbenzene copolymer (PS/DVB), crosslinked polyethyleneglycol, poly-ε-lysine/sebacic acid, controlled pore glass, amino-polyacrylamide resin fiber, cellulose, and hydroxylated polypropylene.

In one embodiment, the one-dimensional core substrate may have a diameter (cross-section) of 10 μm to 100 μm. Specifically, it may be, but is not limited to, 10 μm to 100 μm, 20 μm to 100 μm, 30 μm to 100 μm, 40 μm to 100 μm, 10 μm to 80 μm, 20 μm to 80 μm, 30 μm to 80 μm, 40 μm to 80 μm, 10 μm to 60 μm, 20 μm to 60 μm, 30 μm to 60 μm, 40 μm to 60 μm, 10 μm to 50 μm, 20 μm to 50 μm, 30 μm to 50 μm or 40 μm to 50 μm. If the diameter of the above one-dimensional core substrate is less than 10 μm, it may become too thin, causing the core substrate to clump together and resulting in poor mechanical properties. If it exceeds 100 μm, the loading density of the functional coating may become excessively low.

In one embodiment, the two-dimensional core substrate may have a thickness (layer thickness) of 10 μm to 10 mm. Specifically, the two-dimensional core substrate has a thickness of 10 μm to 10 mm, 50 μm to 10 mm, 100 μm to 10 mm, 150 μm to 10 mm, 200 μm to 10 mm, 250 μm to 10 mm, 300 μm to 10 mm, 350 μm to 10 mm, 400 μm to 10 mm, 10 μm to 5 mm, 50 μm to 5 mm, 100 μm to 5 mm, 150 μm to 5 mm, 200 μm to 5 mm, 250 μm to 5 mm, 300 μm to 5 mm, 350 μm to 5 mm, 400 μm to 5 mm, 10 μm to 3 mm, 50 μm to 3 mm, 100 μm to 3 mm, 150 μm to 3 mm, 200 μm to 3 mm, 250 μm to 3 mm, 300 μm to 3 mm, 350 μm to 3 mm, 400 μm to 3 mm, 10 μm to 1 mm, 50 μm to 1 mm, 100 μm to 1 mm, 150 μm to 1 mm, 200 μm to 1 mm, 250 μm to 1 mm, 300 μm to 1 mm, 350 μm to 1 mm, 400 μm to 1 mm, 10 μm to 0.5 mm, 50 μm to 0.5 mm, 100 μm to 0.5 mm, 150 μm to 0.5 mm, 200 μm to 0.5 mm, 250 μm to 0.5 mm, It may be, but is not limited to, 300 μm to 0.5 mm, 350 μm to 0.5 mm, or 400 μm to 0.5 mm. If the thickness of the two-dimensional core substrate is less than 10 μm, it is difficult to maintain mechanical properties, and if it exceeds 10 mm, there is a possibility that the polymerization reaction may not occur uniformly throughout the interior of the functional coating.

In one embodiment, the porosity of the core substrate may be 50% to 95%. When the core substrate is one-dimensional, core substrates having a shape such as fibers may form pores through entanglement, and when the core substrate is two-dimensional or three-dimensional, it may include pores inside, such as nonwoven fabrics or foams. In cases where the core substrate has pores as such, the porosity of the core substrate may be, but is not limited to, 50% to 95%, 60% to 95%, 70% to 95%, 80% to 95%, 90% to 95%, 50% to 90%, 60% to 90%, 70% to 90%, or 80% to 90%. Here, there is no particular limitation on the pore size of the porous membrane substrate, and if the porosity is less than 50%, the pores may be blocked even after the functional coating swells, preventing the synthesis of biological polymers into the interior of the functional coating. In addition, if the porosity exceeds 95%, it may be difficult to maintain mechanical properties.

In one embodiment, the thickness of the functional coating may be from 0.1 μm to 1000 μm. The thickness of the functional coating may be selected to an appropriate thickness depending on the type of the desired biological polymer and whether the core substrate has a one-dimensional, two-dimensional, or three-dimensional shape. Specifically, the thickness of the functional coating is 0.1 μm to 1000 μm, 0.5 μm to 1000 μm, 1 μm to 1000 μm, 5 μm to 1000 μm, 10 μm to 1000 μm, 50 μm to 1000 μm, 100 μm to 1000 μm, 0.1 μm to 500 μm, 0.5 μm to 500 μm, 1 μm to 500 μm, 5 μm to 500 μm, 10 μm to 500 μm, 50 μm to 500 μm, 100 μm to 500 μm, 0.1 μm to 200 μm, 0.5 μm to 200 μm, 1 μm to 200 μm, 5 μm to 200 μm, 10 μm to 200 μm, 50 μm to 200 μm, 100 μm to 200 μm, 0.1 μm to 100 μm, 0.5 μm to 100 μm, 1 μm to 100 μm, 5 μm to 100 μm, 10 μm to 100 μm, 50 μm to 100 μm, 0.1 μm to 50 μm, 0.5 μm to 50 μm, 1 μm to 50 μm, 5 μm to 50 μm, 10 μm to 50 μm, 0.1 μm to 20 μm, 0.5 μm to 20 μm, 1 μm to 20 μm, 5 μm to 20 μm or 10 μm to 20 The thickness of the functional coating may be, but is not limited to, μm. If the thickness of the functional coating is excessively small, such as less than 0.1 μm, a problem occurs in that the loading density of the functional coating becomes very low. In addition, if the thickness of the functional coating is excessively large, such as exceeding 1000 μm, the functional coating itself may be manufactured through a non-uniform polymerization reaction, and a problem in that the purity decreases during the biological polymer synthesis process may occur.

The above composite solid support may not have a shell-core structure or a bead structure. Additionally, the solid support may not include a core in the shell-core structure.

In one embodiment, the core substrate may have a surface that is hydrophilically treated. Specifically, the surface hydrophilic treatment may be performed by a method selected from a chemical method using a surfactant or an acidic solution and a physical method including plasma treatment or UV irradiation, but is not limited thereto. The functional coating may be formed on the core substrate having a surface that is hydrophilically treated.

In one embodiment, the functional coating may further include a linker.

In one embodiment, the linker may include at least one selected from a Rink amide linker, a Wang linker, a 2-Chlorotrityl (CTC) linker, a Sieber linker, a BAL linker, a 4-sulfamylbutyryl linker, and a HMBA (TFA stabilized) linker.

In one embodiment, the functional coating has a functional group exposed on the inside and/or surface, and the linker can be bonded to the functional group. By adding an additional monomer unit (amino acid residue) to the monomer unit (amino acid residue) linked through the functional coating (functional group)-linker, a biological polymer comprising a plurality of monomer units can be synthesized.

In one embodiment, the biological polymer may include, but is not limited to, one or more selected from a peptide, an oligonucleotide, and a peptide nucleic acid (PNA).

FIG. 1 is a diagram illustrating a biological polymer synthesis system according to one embodiment of the present invention.

Referring to FIG. 1, a biological polymer synthesis system (1) according to one embodiment of the present invention can synthesize a biological polymer by forming a circulating flow in a fluid including at least one of an amino acid, a reagent, and an additive.

Hereinafter, the 'amino acid mixture' as used herein is a liquid comprising amino acid(s) as well as reagent(s), wherein the 'amino acids' may be modified or non-modified and optionally may be pre-activated, and the amino acid mixture may also comprise peptides.

Hereinafter, 'SPPS' is an abbreviation for Solid Phase Peptide Synthesis, and is used to mean producing peptides by adding amino acid residues to peptides or amino acids fixed on a solid support (resin).

Hereinafter, 'reagent' is used to mean coupling reagent, deprotecting reagent, additive, base, and other reagents used in the synthesis.

Hereinafter, 'fluid' is defined as a substance discharged from a storage tank and moving along the piping of a biological polymer synthesis system, and may be a single substance or a mixed substance of various types depending on the process.

As one example, the biological polymer synthesis system (1) can be applied with a flow-through process for solid phase peptide synthesis (SPPS) that enables sequential synthesis of at least one amino acid.

In one embodiment, the biological polymer synthesis system (1) may include a mixing tank (100), a pump (200), a temperature controller (300), and a reactor (400). In addition, the biological polymer synthesis system (1) may further include a valve (500), a solvent tank (500), a sensor (SE), and a detector (DE).

The main line (ML) can provide a circulation pipeline by connecting a mixing tank (100), a pump (200), a temperature controller (300), and a reactor (400). In addition, a valve (500) is arranged in the main line (ML), and the valve (500) can be connected to a first branch line (SL1) and a second branch line (SL2).

The mixing storage tank (100) can provide a space where a fluid containing at least one of an amino acid, a reagent, and an additive stays. The mixing storage tank (100) has an on-off valve (not shown) arranged at the outlet end to control the type and flow rate of the fluid flowing into the main line (ML).

The mixing tank (100) can mix amino acids, reagents, and additives that are introduced from one or more reagent tanks (not shown) and solvent tanks. The operator can precisely control the type and flow rate of the fluid discharged from the mixing tank (100) according to each synthesis step by controlling the opening amount of each chamber. As an example, in each synthesis step of the biological polymer of the SPPS method below, the corresponding fluid can be introduced into the mixing tank, mixed and suspended, and then introduced into the reactor.

1) Deprotection step: Deprotection reagent (piperidine)

2) Washing step: Solvent (DMF)

3) Coupling step: Amino acid, coupling reagent (DIC), additive (Oxyma pure)

4) Washing step: Solvent (DMF)

The pump (200) provides a driving force to the fluid so that the fluid can move along the main line (ML). The pump (200) can provide a driving force so that the fluid flows from the mixing storage tank (100) to the reactor (400). In FIG. 1, the pump (200) is positioned between the mixing storage tank (100) and the reactor (400) in the flow of the fluid, but since the fluid forms a circulating flow, it is not necessarily required to be positioned between the mixing storage tank (100) and the reactor (400).

The pump (200) may be any type of device that provides suction and discharge to the fluid. For example, the pump (200) may be a gear pump, a screw pump, a vane pump, a cap pump, a piston pump, a plunger pump, a diaphragm pump, a centrifugal pump, etc.

In addition, the pump (200) may be any type of pump, such as a mechanical displacement micropump and an electromagnetic motion micropump. A mechanical displacement micropump is a pump that uses the movement of a solid or fluid, such as a gear or diagram, to generate a pressure difference to induce a fluid flow, and includes a diaphragm displacement pump, a fluid displacement pump, a rotary pump, etc. An electromagnetic motion micropump is a pump that uses electric or magnetic energy to directly move a fluid, and includes an electrohydrodynamic pump (EHD), an electroosmotic pump, a magnetohydrodynamic pump, an electrowetting pump, etc.

The temperature controller (300) can set the temperature of the fluid flowing into the reactor (400) to a target temperature. The temperature controller (300) is arranged downstream of the mixing storage tank (100) and upstream of the reactor (400) to cool or heat the fluid including at least one selected from amino acids, reagents, and additives. In FIG. 1, the temperature controller (300) is positioned between the mixing storage tank (100) and the reactor (400) in the flow of the fluid, but since the fluid forms a circulating flow, it does not necessarily have to be positioned between the mixing storage tank (100) and the reactor (400).

The temperature controller (300) may be a variety of devices that transfer heat to a fluid or receive heat from a fluid through heat exchange. For example, the temperature controller (300) may be an electric heating device, an induction heater, or a microwave cavity.

As an example, the temperature controller (300) can maintain the temperature of the fluid flowing into the reactor (400) at room temperature. The temperature controller (300) can control the temperature of the fluid to 120° C. or lower, specifically, to 25° C. to 100° C. In addition, the temperature controller (300) can control the temperature of the fluid to 50° C. to 80° C.

Figure 2 is a drawing illustrating the reactor of Figure 1.

Referring to FIGS. 1 and 2, the reactor (400) can circulate and flow the fluid discharged from the mixing tank (100).

The reactor (400) may have a column (410) and a composite solid support (420).

The column (410) may have an inlet (411) through which a fluid is introduced and an outlet (412) through which a fluid is discharged. The column (410) has an internal space, and a composite solid support (420) may be arranged therein.

The column (410) may have various sizes depending on the type of biological polymer being synthesized, the amount of biological polymer being synthesized, the flow rate of the fluid, the velocity of the fluid, the type of the composite solid support, etc.

The inlet (411) and outlet (412) of the column (410) are arranged on the main line (ML), so that the fluid entering the inlet (411) can pass through the internal space of the column (410) and be discharged through the outlet (412). At this time, the fluid can pass through the composite solid support (420) arranged in the internal space of the column (410).

A composite solid support (420) can be placed inside the column (410). The solid support (420) can be filled inside the column (410).

The composite solid support (420) can be arranged in various positions and shapes in the internal space of the column (410). The composite solid support (420) can be filled in a preset position in the internal space of the column (410).

For example, a composite solid support (420) can be loaded into the internal space of the column (410).

For example, if the core material of the composite solid support (420) is in the form of entangled filaments (FM), the fluid can pass between the filaments (FM). The flow path of the fluid can vary depending on the method of loading the composite solid support (420). For example, if a composite solid support in a spirally rolled form is loaded, the fluid can flow between each layer of the spiral composite solid support and in the porous space generated by the entanglement.

The composite solid support (420) may have a core substrate (BM) and a functional coating (CO).

As an example, the core substrate (BM) may be formed by entangling multiple filaments to form random porosity. The functional coating (CO) may be a polymer for synthesizing biological polymers coated on the core substrate so that functional groups such as amine groups and carboxyl groups are exposed.

The core substrate (BM) may have a length of 1 mm or more and a shape of one or more dimensions. The core substrate (FM) may have one or more shapes selected from a one-dimensional shape, a two-dimensional shape, and a three-dimensional shape.

The core substrate (BM) may have solvent resistant, thermal resistant, or both of the above properties. Unlike the functional coating (CO), the core substrate (BM) does not have swelling properties or solvent-soluble properties, and thus is capable of self-standing and can act as a support for a composite solid support. In addition, the core substrate (FM) has thermal resistance, and thus can act as a support without being changed even when the synthesis of a biological polymer is carried out at a high temperature.

A functional coating can surround a core substrate (FM) and provide an area where biological polymers can be synthesized.

In one embodiment, the functional coating (CO) may have the property of swelling in a solvent.

The functional coating (CO) may include functional groups on the surface and inside. Specifically, the functional coating includes a functional group, and due to the swelling characteristic in a solvent, it is easy for a reactant to penetrate into the functional coating due to a concentration gradient. As a result, the functional groups on the surface and inside of the functional coating are easily exposed to the reactant, so that high synthesis efficiency can be exhibited even at a low loading density.

The valve (500) is arranged between the reactor (400) and the storage tank (100) to set a path for discharging waste. The valve (500) can control the flow direction of the fluid so that the waste generated after the reaction of the reactor (400) is discharged to the second branch line (SL2). As an example, the valve (500) can be a four-channel valve, and the four-channel valve can be connected to the mixing tank (100), the reactor (400), the solvent storage tank (600), and the second branch line (SL2).

The valve (500) can control the circulation of the fluid and the discharge of waste. The valve (500) can set the circulation flow of the reactant fluid during each stage of SPPS synthesis to increase the synthesis rate of the biological polymer, and can discharge the remaining material of each stage to the second branch line (SL2) to enable the process to proceed to the next stage.

The solvent storage tank (600) can store solvents used for biological polymer synthesis and cleaning. The solvent can be selected in various ways depending on the synthesis process.

The solvent storage tank (600) may be placed on the first branch line (SL1). The solvent storage tank (600) is connected to the first branch line (SL1), so that the solvent may be moved to the valve (500) or the mixing storage tank (100) according to each stage of the SPPS synthesis.

Before the washing process is performed during the synthesis process, the substances remaining in the pipes in the previous step must be removed. At this time, the solvent discharged from the solvent storage tank (600) is discharged to the first branch line (SL1), so that the main line (ML) can be washed.

The sensor (SE) is arranged on the main line (ML) and can sense at least one of temperature, pressure, and flow rate of the fluid flowing along the main line (ML).

The detector (DE) is arranged on the main line (ML) and can detect the progress of the biological polymer synthesis. For example, the detector can be arranged at least one of the inlet and the outlet of the reactor (400) and can detect the circulating fluid. In Fig. 1, the detector (DE) is arranged between the reactor (400) and the mixing reservoir (100) in the fluid flow, but since the fluid forms a circulating flow, it does not necessarily have to be arranged between the reactor (400) and the mixing reservoir (100).

The main line (ML) can connect a mixing tank (100), a pump (200), a temperature controller (300), and a reactor (400), and can provide a circulation path for the fluid.

The first branch line (SL1) branches from the main line (ML) between the mixing storage tank (100) and the reactor (400) and can be connected to the mixing storage tank (100). The first branch line (SL1) connects the valve (500) and the mixing storage tank (100) to supply the solvent in the solvent storage tank (600) to the main line (ML), thereby removing the substances remaining in the main line (ML).

The second branch line (SL2) can be branched from the valve (500) to discharge waste.

Hereinafter, preferred examples are presented to help understand the present invention. However, the following examples are provided only to help understand the present invention more easily, and the content of the present invention is not limited by the following examples.

Example 1. Analysis of pressure drop in reactor according to flow rate

Figure 3 is a graph measuring pressure drop according to flow rate to compare the reactor of the present invention with a comparative example.

(Test equipment)

- Column (Intertec empty cartridge column): 27 mL, I.D./Hight: 12.8*60 mm

- Pump: LEAD FLUID CT3001, Pump head: Fluid-o-Tech/MG209

- Reagents used: DMF (Dimethylformaminde, SAMJHUN), Rink-Amide-MBHA-Resin (GL Biochme, 0.5mmole/g)

- Pressure measurement equipment: SMC PSE560-C01 pressure sensor, SMC PSE200A digital pressure sensor controller

As an example, 5.0 g (0.5 mmol, loading density: 0.1 mmol/g) of a composite solid support was placed in the column, and as a comparative example, 1.0 g (0.5 mmol, loading density: 0.5 mmol/g) of a particle-type solid support (PS/DVB) was placed in the column, and then 20 mL of DMF solvent was added and the composite solid support was swollen for 30 minutes.

Afterwards, the discharge tube of the pump was connected to the column, pressure sensors were installed at the inlet and outlet of the column, and each pressure sensor was connected to the controller.

DMF was flowed at a flow rate of 40 mL/min (1.48 C.V/min) through an empty column, a composite solid support column (Example), and a particle type solid support column (Comparative Example), and the pressures at the inlet and outlet were measured. Thereafter, the pressures at the inlet and outlet of the columns were measured while changing the flow rates to 50 ml/min (1.85 C.V/min), 100 ml/min (3.7 C.V/min), 200 ml/min (7.4 C.V/min), 300 ml/min (11.11 C.V/min), 400 ml/min (14.81 C.V/min), and 500 ml/min (18.52 C.V/min).

The pressure drop values that occurred at the inlet and outlet of the empty column were set as the reference value (Zero). The additional pressure drop values that occurred were calculated by comparing them with the reference values at the inlet and outlet of the column containing the composite solid support, and the additional pressure drop values that occurred were calculated by comparing them with the reference values at the inlet and outlet of the column containing the particle-type solid support.

Flow rate (ml/min) Comparative example Example 40 0.17 0.04 50 0.23 0.04 100 0.5 0.05 200 0.97 0.08 300 - 0.1 400 - 0.11

Referring to FIG. 3 and Table 1, the composite solid support of the present invention exhibits a small pressure drop even when the flow rate increases. This is because the fluid can pass through the empty space of the composite solid support, so even when the flow rate increases, the pressure drop occurs small and the fluid flowability increases. In contrast, the conventional particle-type solid support (bead-type solid-phase synthetic resin polymer (PS/DVB)) exhibits a large pressure drop when the flow rate increases and the fluid is not discharged at 300 ml/min. This is because the conventional particle-type solid support clumps together when the flow rate increases to fill the space within the column, reducing the space through which the fluid can pass, significantly reducing the pressure measured at the outlet, and decreasing the fluid flowability.

Example 2. Pressure drop analysis of composite solid support according to column size

Figure 4 is a graph measuring the pressure drop according to the size of the reactor of the present invention.

(Test equipment)

- A column (Intertec empty cartridge column): 27 mL, I.D./Hight: 12.8*60 mm

- B column (Intertec empty cartridge column): 108 mL, I.D./Hight: 21.4*76 mm)

- C column (Intertec empty cartridge column): 385mL, I.D./Hight: 26.8*127 mm

- Pump: LEAD FLUID CT3001, Pump head: Fluid-o-Tech/MG209

- Reagents used: DMF (Dimethylformaminde, SAMJHUN), Rink-Amide-MBHA-Resin (GL Biochme, 0.5mmole/g)

- Pressure measurement equipment: SMC PSE560-C01 pressure sensor, SMC PSE200A digital pressure sensor controller

Column A was loaded with 5 g of a composite solid support, 20 ml of DMF solvent was added, and the mixture was swollen for 30 minutes. Column B was loaded with 20 g of a composite solid support, 80 ml of DMF solvent was added, and the mixture was swollen for 30 minutes. Column C was loaded with 70 g of a composite solid support, 300 ml of DMF solvent was added, and the mixture was swollen for 30 minutes.

The discharge tube of the pump was connected to each column, a pressure sensor was installed at the inlet and outlet of each column, and each pressure sensor was connected to a controller.

DMF was flowed at a flow rate of 0.5 column volume (mL)/min through columns A, B, and C, respectively, and the pressures at the inlet and outlet were measured. After that, the flow rates were changed to 1 C.V/min, 2 C.V/min, 3 C.V/min, 4 C.V/min, and 5 C.V/min, and the pressures at the inlet and outlet of the columns were measured.

The pressure drop values that occurred at the inlet and outlet of each empty column were set as the reference value (Zero). The additional pressure drop values that occurred at the inlet and outlet of columns A, B, and C containing composite solid supports were calculated by comparing them with the reference values.

Flow rate (C.V/min) A column Column B C column 0.5 0.03 0.05 0.05 1 0.04 0.06 0.05 2 0.04 0.06 0.07 3 0.04 0.07 0.08 4 0.05 0.08 0.09

Referring to FIG. 4 and Table 2, the composite solid support of the present invention exhibits a small pressure drop in the fluid at various flow rates even when the volume of the column is varied. This is because the fluid can pass through the empty space of the composite solid support, so that the pressure drop is small and the fluidity of the fluid increases even when the flow rate increases. Accordingly, the composite solid support of the present invention exhibits a consistently small degree of pressure drop regardless of the volume of the column.

Example 3. Pressure drop analysis of composite solid supports according to temperature

Figures 5 and 6 are graphs measuring the pressure drop according to temperature in the reactor of the present invention and a comparative example.

When the rotation speed of the same pump is fast at the same flow rate, the back pressure is large. Through this, the rotation speed of the pump when a specific flow rate is discharged was measured and the back pressure applied to each solid support was compared.

A 27 ml column (Example) loaded with the composite solid support (0.5 mmol) used in the present invention, an empty column (Comparative Example 1), and a 27 ml column (Comparative Example 2) loaded with a conventional particle-type solid support (Rink-Amide-MBHA-Resin, 0.5 mmol, 1.25 g) were compared.

As shown in Table 3 and Fig. 5, the RPM values of the pump were recorded when flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 ml/min were achieved using DMF solvent at 25°C.

When DMF was used at 25°C, the pump rotation speed of the column loaded with the composite solid support (Example) did not differ significantly from that of the empty column (Comparative Example 1). On the other hand, the pump operation of the column loaded with the particle-type solid support (Comparative Example 2) stopped at 250 ml/min due to high back pressure.

Flow rate (ml/min) Example Comparative Example 1 Comparative Example 2 50 96 94 103 100 191 188 205 150 287 282 317 200 383 376 542 250 479 471 0 300 574 565 0 350 670 646 0 400 766 724 0

As shown in Table 4 and Fig. 6, the RPM values of the pump were recorded when the flow rates were 50, 100, 150, 200, 250, 300, 350, and 400 ml/min using DMF solvent at 50°C. When DMF was used at 50°C, the viscosity of the solution decreased, which reduced the back pressure. However, in the case of the particle-type solid support (Comparative Example 2), the pump stopped operating at 350 ml/min due to the high back pressure.

Flow rate (ml/min) Example Comparative Example 1 Comparative Example 2 50 92 89 97 100 184 177 194 150 276 266 378 200 368 355 474 250 459 443 554 300 551 532 768 350 669 621 0 400 765 710 -

As shown in Table 5, the RPM values of the pump were recorded when the flow rates were 50, 100, 150, 200, 250, 300, 350, and 400 ml/min using DMF solvent at 70℃. When DMF was used at 70℃, the viscosity of the solution decreased, which reduced the back pressure. However, in the case of the particle-type solid support (Comparative Example 2), the pump stopped operating at 350 ml/min due to the high back pressure.

Flow rate (ml/min) Example Comparative Example 1 Comparative Example 2 50 89 87 94 100 180 175 189 150 271 259 375 200 364 349 465 250 453 440 543 300 547 530 730 350 659 618 0 400 763 705 -

In addition, the rotation speed of the pump was measured at room temperature using Piperidine 20% in DMF (0.1 M Oxymapure), which is widely used in solid-phase peptide synthesis reactions. At this time, the particle-type solid support stopped operating due to high back pressure at 250 ml/min. That is, it was confirmed that the composite solid support of the present invention was subjected to less back pressure than the conventional particle-type solid support. Considering that the volume and weight of the composite solid support increase as a longer peptide is synthesized, resulting in more back pressure, the composite solid support of the present invention can enhance the synthetic performance of a flow reactor by increasing the fluid flowability, unlike the conventional particle-type solid support.

A flow reactor according to one embodiment of the present invention and a biological polymer synthesis system including the same can increase the efficiency of a synthesis reaction.

Conventional particle-type solid-phase synthetic resin polymers are aggregated at high flow rates and fill the internal space of the column without any empty space, which reduces the flowability of the reactor. As a result, the reactor experiences a high pressure drop at the outlet, and accordingly, a high back pressure occurs. A high back pressure cannot significantly increase the synthesis scale of the synthesis system.

According to one embodiment of the present invention, a flow reactor and a biological polymer synthesis system including the same have increased flowability of the reactor by a composite solid support. Accordingly, the reactor has a low pressure drop and thus achieves a low back pressure, so that an increase in the synthesis scale is possible.

Specifically, under the same pressure conditions, when the back pressure is low, the rate of decrease (loss) in the flow rate is small, so mixing and diffusion between the reactants and the reaction solution within the column are promoted. The biological polymer synthesis system has improved flow dynamics as the flow rate increases, and the reaction efficiency and yield are improved.

In addition, when the back pressure is reduced, the wear and mechanical stress of the internal parts of the reactor are reduced, which improves the durability of the entire system. In addition, since pumps that provide various pressures can be applied, the usability and range of selection can be increased. In particular, even when using a low-pressure pump, high reaction efficiency and yield can be achieved.

Additionally, reducing back pressure reduces the risk of damage to pipes and connections through which the fluid flows, thereby reducing fluid leakage and improving the durability of the system and the stability and reliability of the process.

In addition, since the fluid passing through the reactor can be precisely controlled when the back pressure is reduced, the reproducibility of the reaction process is improved, and consistent quality and high-quality biological polymers can be synthesized.

Example 4. Preparation of composite solid support

(1) Manufacturing of polymers for functional coating

An experiment was conducted to measure the performance of synthesizing biological polymers using the previously confirmed flow reactor. First, a functional coating polymer used for a composite solid support was prepared. Each reagent as shown in Table 6 below was used, and each reagent was uniformly dissolved by mixing and stirring in a solvent. At this time, the components of the functional coating polymer should satisfy each mass ratio (weight part) range, and the solvent can be used regardless since it will be dried later. In the experiment, the sum of each reagent and solvent (D.W) was measured to be 100 g [the solvent can be used within 10 wt% to 90 wt% of the total (reagent + solvent)].

classification Reagent name Mass part First monomer Polyethylene Glycol Diacrylate 100 Second monomer Methyl acrylate 0~50 Active monomer N-(2-aminopropyl) methacrylamide hydrochloride 1~25 Surfactant Sodium Dodecyl Sulfate 0~25

After that, 0.5 g of 2-Hydroxy-2-methylpropiophenone, a curing reaction initiator, was added to the solution and mixed uniformly to prepare a functional coating solution. The functional coating solution was uniformly applied to a petri dish and sufficiently irradiated with ultraviolet energy of 10 mJ/cm2 to 1,000 mJ/cm2 to ensure curing. After curing was complete, the reaction residue was removed using ethanol to obtain a functional coating polymer disk. The mass of the functional coating polymer after drying was measured to be 45.7 g. The manufactured functional coating polymer was rapidly frozen using liquid nitrogen and then pulverized using a mortar and pestle. Thereafter, a 100, 200 mesh sieve was used to obtain functional coating polymer particles having a particle size range of 100-200 mesh (74-149 μm).

(2) Preparation of composite solid support for biological polymer synthesis

In order to manufacture a composite solid support, the functional coating solution (before curing) of (1) of the above Example 4 and 20 g of a core substrate (polypropylene, spunbond nonwoven fabric, 40 g/m ² ) were prepared. The core substrate was sufficiently impregnated in the functional coating solution, and after the core substrate was taken out of the functional coating solution, it was sufficiently irradiated with ultraviolet energy of 10 mJ/cm ² to 1,000 mJ/cm ² in an ultraviolet curing chamber.

After the curing process was completed, the coated core substrate was subjected to a process of removing reaction residues using ethanol and drying. A composite solid support in which a functional coating polymer was coated on a core substrate was obtained (solid support 30.4 g, 60.8 g/m ² ). The appearance of the core substrate before coating is as shown in Fig. 7, and the appearance of the composite solid support is as shown in Fig. 8.

Example 5. Synthesis of biological polymers using composite solid supports

In comparison with the method of a batch-type automatic synthesizer (CEM Liberty, USA), it was investigated whether peptide synthesis is possible in the case of SPPS synthesis using a biological polymer synthesis system including a column-type flow reactor containing a solid support for biological polymer synthesis according to one specific example, and whether there is a difference in the purity and yield of the synthesized peptide.

(1) Synthesis of peptides using a batch-type automatic synthesizer

After adding 0.1 mmol of each of the existing solid-phase synthesis particle-type solid support and the composite solid support according to one specific example to an automatic synthesizer, the synthesis reaction of the ACP (acyl carrier protein) model peptide (65-74) of the following sequence number 1 was performed under the same conditions as in Table 7 below.

Sequence number 1: VQAAIDYING

Reaction stage reagent
(Solvent: DMF) Volume (mL) Temperature (℃) Time (minutes) Deprotection 20% piperidine,0.1M Oxyma pure 15 70 2 purifying DMF 10 (3 times) - - Coupling Fmoc-Ile(0.2M) 6 70 4 DIC (0.5M) 2 Oxyma pure(1.0M) 1 purifying DMF 10 (2 times) - -

After the above peptide synthesis reaction was completed, the particle-type solid support and the composite solid support were each sufficiently washed with ethanol and dried, and a cleavage reaction was performed for 2 hours using a stripping solution (95% TFA, 2.5% TSI, 2.5% DW). After that, the solid was precipitated using cold ether and centrifuged to obtain a solid, which was washed twice more using 15 ml of cold ether, and then sufficiently dried using reduced pressure drying. The purity and yield of the dried peptide were measured using high-performance liquid chromatography (HPLC). The results of measuring the purity and yield of the dried peptide are shown in Table 8.

Types of solid supports water(%) transference number(%) Composite solid support 85.1 88.4 Particulate solid support 81.4 71.5

In the analysis of peptide synthesis using a particulate solid support and a biological polymer composite solid support according to one specific example using a conventional commercial batch-type SPPS synthesizer (Liberty, CEM, USA), the yield and purity of ACP peptides were shown to be somewhat improved in the composite solid support according to one specific example than in the particulate solid support.

(2) Peptide synthesis using a flow-based column synthesizer

In order to compare the synthetic performance of a particulate solid support and a homogeneous composite solid support in a flow reactor, 0.5 mmol of each of a particulate solid support and a composite solid support was added to a 27 mL column, and the reaction step was performed by changing the types of amino acids under the conditions shown in Table 9 below to synthesize the acyl carrier protein (ACP) model peptide (65-74) of the sequence number 1. After that, the dry mass was measured to calculate the yield, and the purity of the synthesized peptide was measured using high-performance liquid chromatography (HPLC).

Reaction stage reagent
(Solvent: DMF) Volume (mL) temperature
(℃) hour
(minute) flux
(mL/min) Deprotection 20% piperidine,
0.1M Oxyma pure 25 70 2 100 purifying DMF 125 - - Coupling Fmoc-Amino acid(0.2M) 15 70 4 DIC (0.5M) 6 Oxyma pure(1.0M) 3 purifying DMF 20 - -

The synthetic performance in a flow reactor using a composite solid support was shown to have a purity of 95.8% and a yield of 93.5%, which were superior to those in a batch reactor in terms of both synthetic purity and yield. In contrast, when a flow reactor was filled with a particulate solid support to synthesize ACP(65-74) peptide, a backpressure issue occurred during the synthesis, which prevented the synthesis from proceeding properly. This is because the particulate solid support blocks the column in the flow reactor due to swelling, and the composite solid support, which exists in a state where a functional coating is coated on the core substrate, is less likely to have this problem, indicating that it is more suitable for use in a flow reactor. Through this, it was confirmed that the composite solid support is suitable for both batch and flow-type synthesis systems compared to the existing particulate solid support, and in particular, when applied to a flow-type synthesis system, the yield and purity of the ACP peptide were over 90%, showing significantly superior efficacy in the synthesis of biological polymers.

Example 6. Analysis and evaluation of changes in yield and purity of synthesized peptides according to the flow rate of biological polymers in a flow reactor

As in Example 5, the yield and purity of the synthesized peptide according to the flow rate were confirmed using a composite solid support. ACP was synthesized using the same method as in Example 5, and the results of the purity and yield of the synthesized ACP peptide are shown in Table 10 and Figures 9 to 11 below.

Flow rate (mL/min) transference number(%) water(%) 100 84.9 85.3 200 88.3 90.5 300 91.1 95.7

Figures 9 to 11 are composite solids for synthesizing biological polymers according to one specific example.

This shows an HPLC graph for synthesized ACP according to the flow rate using a support. When a solid support for the synthesis of biological polymers according to one specific example was used, it was confirmed that when the flow rate of the column synthesizer increased, the purity and yield of the peptide increased.

The above results imply that a flow-based reactor including a column is a more suitable reactor for a composite solid support according to one embodiment.

Although the present invention has been described with reference to the embodiments shown in the drawings, these are merely exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible. Therefore, the true technical protection scope of the present invention should be determined by the technical idea of the appended claims.

Claims (14)

A mixing tank in which at least one of amino acids, reagents and additives is mixed; A reactor into which the fluid discharged from the above mixing tank is introduced; A temperature controller arranged between the mixing tank and the reactor to control the temperature of the fluid flowing into the reactor; and a pump providing a driving force to cause the fluid to flow from the mixing tank to the reactor; The above reactor A column having an inlet through which the fluid flows in and an outlet through which the fluid flows out; and A biological polymer synthesis system comprising a composite solid support disposed inside the above column. In the first paragraph, The above composite solid support is Core material; and A biological polymer synthesis system comprising a functional coating positioned on the core substrate. In the second paragraph, The above composite solid support A biological polymer synthesis system that is self-standing when the fluid passes through the column. In the second paragraph, The above composite solid support is A biological polymer synthesis system, wherein the loading density of the functional coating is 0.01 mmol/g to 2 mmol/g. In the first paragraph, A biological polymer synthesis system, wherein the pressure drop between the inlet and the outlet is 10 bar or less at a flow rate of 40 C.V/min or less when the fluid passes through the reactor. In the first paragraph, A biological polymer synthesis system, wherein the temperature of the fluid flowing into the reactor after passing through the temperature controller is 120°C or less. In the first paragraph, A biological polymer synthesis system further comprising a valve disposed between the reactor and the mixing tank to discharge waste. In the first paragraph, A main line connecting the above mixing tank, the pump, the temperature controller and the reactor; and A biological polymer synthesis system further comprising a first branch line branched from the main line between the mixing tank and the reactor and connected to the mixing tank. In Article 8, A biological polymer synthesis system, wherein the fluid has a circulating flow through the main line. In Article 8, A biological polymer synthesis system further comprising a solvent storage tank disposed on the first branch line. In Article 10, The above solvent storage tank A biological polymer synthesis system, connected to the valve and the mixing tank through the first branch line. In Article 8, A biological polymer synthesis system further comprising a sensor disposed on the main line and sensing at least one of temperature, pressure and flow rate of the fluid flowing along the main line. In Article 8, A biological polymer synthesis system further comprising a detector disposed on the main line to detect the progress of synthesis of the biological polymer. In Article 7, A biological polymer synthesis system further comprising a second branch line branched from the valve and discharging the waste.

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