Flow reactor and biological polymer synthesis system comprising the same

Abstract

The present invention relates to a flow reactor and a biological polymer synthesis system comprising the same, comprising a mixing reservoir 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 reservoir is introduced, a temperature controller disposed between the mixing reservoir and the reactor to control the temperature of the fluid introduced into the reactor, and a pump providing a driving force to cause the fluid to flow from the mixing reservoir to the reactor, wherein the reactor comprises a column having an inlet for the fluid to be introduced and an outlet for the fluid to be discharged, and a composite solid support disposed inside the column.

Description

Technology Field

[0001] The present invention relates to a flow reactor and a biological polymer synthesis system. Background Technology

[0002] Peptide pharmaceuticals generate billions of dollars in annual revenue in the diabetes, obesity, and oncology industries, and their scope is expanding into the development of new drugs for emerging diseases such as cardiovascular and neurodegenerative diseases. Currently, the disadvantage of peptides—their short in vivo half-life—has been overcome through methods such as the introduction of non-natural amino acids. Various peptide pharmaceuticals with dosing cycles of more than one week are already on the market, and numerous oral administration candidates are undergoing clinical trials under the U.S. Food and Drug Administration (FDA). Consequently, production demand for peptide pharmaceuticals is increasing by nearly 10% annually, creating an urgent need for new synthesis platforms to improve productivity to meet this demand and to address ESG issues, such as regulations on the use of DMFs used in traditional chemical synthesis methods.

[0003] Peptide drugs are largely manufactured by biological or chemical methods. Biological methods using genetic recombination or transgenic animals and plants have the disadvantage that it is difficult to produce peptide drugs with increased half-lives using non-natural amino acids or PEGylation, which are recent trends, and it is difficult to remove various endogenous contaminants derived from living organisms or exogenous contaminants generated during the process.

[0004] In chemical methods, liquid-phase reactions are inexpensive to process, but they can only synthesize peptides composed of 10 or fewer amino acids, making it generally impossible to produce peptide drugs composed of 20 or more amino acids. Solid Phase Peptide Synthesis (SPPS) is the most suitable method for manufacturing PEGylated peptide drugs composed of 20 to 50 different types of natural / non-natural amino acids.

[0005] To enhance the production capacity of various biological polymers, including peptides, various types of reactors are being researched in the field of SPPS. Representative reactors include batch-type and flow-type reactors. For large-scale synthesis, batch-type reactors have gradually scaled up from 10–50 mL to the current 100–500 L units, with improvements made to various solution transfer, stirring, pressure control, and filtration methods. On a laboratory scale, the recent introduction of flow reactors has demonstrated the ability to rapidly synthesize peptides with high purity and yield.

[0006] However, batch reactors have problems such as differences in temperature transfer between the center and surface of the reaction solution and the time required for uniform mixing due to the reactor size varying according to the reaction scale, as well as difficulties in yield and quality control resulting from low reproducibility of reaction temperature and mixing.

[0007] On the other hand, conventional flow reactors have the advantage of excellent reaction reproducibility, as high-speed mixing through a tubular reactor increases the reaction specific surface area, thereby inducing uniform temperature transfer and chemical reactions, and facilitating quality and yield control due to this high reproducibility.

[0008] While currently developed flow reactors have the advantage of rapid synthesis reactions, there is a problem where back pressure of 30-50 bar occurs when using 200 mg PS / DVB. Such high back pressure clearly demonstrates that the synthesis scale cannot be significantly increased. Additionally, although the flow reactor developed by Vaportek in the UK solved the problem of high back pressure, which is a disadvantage of packed bead resins, by using an adjustable syringe, it has limitations such as a very small usable flow rate and difficulty in significantly increasing the synthesis scale due to the nature of pressure control using a syringe.

[0009] Furthermore, conventional particulate resins have structural limitations due to the inherent limitations of their particle structure, making self-standing impossible; they also have the structural limitation of increasing back pressure within the flow reactor as they move and accumulate due to the flow.

[0010] Accordingly, a self-supporting solid support material suitable for SPPS-type flow reactors and a biological polymer synthesis system utilizing the same were newly developed. The problem to be solved

[0011] The present invention aims to provide an SPPS-type flow reactor having improved synthesis efficiency and yield, as well as high durability and stability, and a biological polymer synthesis system including the same. means of solving the problem

[0012] 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 the temperature of the fluid introduced into the reactor; and a pump providing a driving force to cause the fluid to flow from the mixing tank to the reactor, wherein the reactor comprises a column having an inlet for the fluid to be introduced and an outlet for the fluid to be discharged, and a composite solid support disposed inside the column.

[0013] In addition, the composite solid support may comprise a core substrate and a functional coating located on the core substrate.

[0014] In addition, the composite solid support can self-stand when the fluid passes through the column.

[0015] In addition, the loading density of the functional coating on the composite solid support may be 0.01 mmol / g to 2 mmol / g.

[0016] 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 CV / min or less. Specifically, the pressure drop may be measured based on the pressure difference between the inlet and the outlet of an empty column. In addition, the above flow rate can be expressed as CV (Column Volume) per minute, and the reactor has a flow rate of 40 CV / min or less, specifically 40 CV / min or less, 30 CV / min or less, 20 CV / min or less, 0.1 CV / min to 40 CV / min, 1 CV / min to 40 CV / min, 5 CV / min to 40 CV / min, 10 CV / min to 40 CV / min, 15 CV / min to 40 CV / min, 20 CV / min to 40 CV / min, 25 CV / min to 40 CV / min, 30 CV / min to 40 CV / min, 35 CV / min to 40 CV / min, 0.1 CV / min to 30 CV / min, 1 CV / min to 30 CV / min, 5 CV / min to 30 CV / min, 10 CV / min to 30 CV / min, 15 CV / min to The pressure drop may be 10 bar or less at 30 CV / min, 20 CV / min to 30 CV / min, 25 CV / min to 30 CV / min, 0.1 CV / min to 20 CV / min, 1 CV / min to 20 CV / min, 5 CV / min to 20 CV / min, 10 CV / min to 20 CV / min, 15 CV / min to 20 CV / min, 0.1 CV / min to 10 CV / min, 1 CV / min to 10 CV / min, or 5 CV / min to 10 CV / min.In addition, within the above flow velocity 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 may 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.

[0017] 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 is capable of synthesizing biological polymers with excellent purity and efficiency even in various 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, or 25°C to 50°C, but is not limited thereto, and can be set without limitation as long as it is the temperature of the fluid used for biological polymer synthesis.

[0018] In addition, it may further include a valve disposed between the reactor and the mixing storage tank to discharge waste.

[0019] Additionally, it may further include a main line connecting the mixing storage tank, the pump, the temperature controller, and the reactor, and a first branch line branched from the main line between the mixing storage tank and the reactor and connected to the mixing storage tank.

[0020] In addition, the fluid may have a circulating flow through the main line.

[0021] In addition, it may further include a solvent storage tank disposed on the first branch line.

[0022] In addition, the solvent storage tank can be connected to the valve and the mixing storage tank via the first branch line.

[0023] Additionally, it may further include a sensor positioned on the main line to sense at least one of the temperature, pressure, and flow rate of the fluid flowing along the main line.

[0024] In addition, it may further include a detector positioned on the main line to detect the progress of the synthesis of the biological polymer.

[0025] In addition, it may further include a second branch line that branches off from the valve and discharges the waste. Effects of the invention

[0026] 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 utilizes a flow reactor loaded with a composite solid support, which is characterized by exhibiting a low pressure drop even at high flow rates and maintaining excellent efficiency and yield over various fluid temperature ranges.

[0027] In addition, the flow reactor of the present invention and the biological polymer synthesis system including it improve the overall durability and stability of the equipment and enable precise process control. Brief explanation of the drawing

[0028] FIG. 1 is a drawing 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 showing the pressure drop according to the flow rate to compare the reactor of the present invention with a comparative example. Figure 4 is a graph showing the pressure drop according to the size of the reactor of the present invention. Figures 5 and 6 are graphs showing the pressure drop according to temperature of 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. FIGS. 9 to 11 show HPLC graphs of ACP synthesized according to flow rate using a composite solid support according to one embodiment. Specific details for implementing the invention

[0029] The present invention is capable of various modifications and may have various embodiments; 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 by referring to the embodiments described below in detail together with the drawings. However, the present invention is not limited to the embodiments disclosed below but can be implemented in various forms.

[0030] 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 reference numerals, and redundant descriptions thereof will be omitted.

[0031] In the following embodiments, terms such as first, second, etc. are used not in a limiting sense, but for the purpose of distinguishing one component from another component.

[0032] In the following examples, singular expressions include plural expressions unless the context clearly indicates otherwise.

[0033] In the following embodiments, terms such as "include" or "have" mean that the features or components described in the specification are present, and do not preclude the possibility that one or more other features or components may be added.

[0034] In the following embodiments, when a part such as a film, region, or component is described as being on or above another part, it includes not only the case where it is directly on top of the other part, but also the case where another film, region, or component is interposed in between.

[0035] In the drawings, the size of components may be exaggerated or reduced for convenience of explanation. For example, the size and thickness of each component shown in the drawings are depicted arbitrarily for convenience of explanation, so the present invention is not necessarily limited to what is illustrated.

[0036] In the following embodiments, when a membrane, region, component, etc. is described as being connected, it includes not only cases where the membrane, region, or component is directly connected, but also cases where other membranes, regions, or components are interposed between them to form an indirect connection. For example, when a membrane, region, component, etc. is described as being electrically connected in this specification, it includes not only cases where the membrane, region, or component, etc. are directly electrically connected, but also cases where other membranes, regions, or components are interposed between them to form an indirect electrical connection. Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings.

[0037] In this specification, the term "loading density" means "the number of moles of functional groups provided per unit mass of a composite solid support," and may refer to the synthesis reaction sites of a biological polymer according to the compositional ratio of a polymer for biological polymer synthesis according to one embodiment.

[0038] In one embodiment, the composite solid support comprises a core substrate (Structure core) 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.

[0039] In one embodiment, the functional coating may have the characteristic of swelling in a solvent.

[0040] In one embodiment, the functional coating may include functional groups on its surface and inside. Specifically, the functional coating includes functional groups, and due to its swelling characteristic in a solvent, it facilitates the penetration of reactants into the functional coating due to a concentration gradient. As a result, the functional groups on the surface and inside the functional coating are easily exposed to the reactants, thereby enabling high synthesis efficiency even at low loading densities.

[0041] In one embodiment, the functional group may include one or more selected from amine groups, carboxyl groups, hydroxyl groups, carbonyl groups, amino groups, thiol groups, and phosphate groups.

[0042] In one embodiment, the loading density of the functional coating may be 0.01 mmol / g to 2 mmol / g. Specifically, the reaction site loading density 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, 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.It may be 7 mmol / g to 1 mmol / g, but is not limited thereto.

[0043] 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 may have different expansion rates per unit mass depending on the type of solvent, as swelling occurs in solvents other than water.

[0044] In one embodiment, the core substrate may possess solvent resistance, thermal resistance, or both characteristics. Unlike the functional coating, the core substrate does not have swelling characteristics or dissolution in a solvent, and thus can self-stand even at high flow rates and serve as a support for a composite solid support. Furthermore, the core substrate possesses thermal resistance, so it can serve as a support without being altered even if the synthesis of the biological polymer proceeds at high temperatures.

[0045] In one embodiment, the components of the core substrate may include one or more selected from polymers, metals, and ceramics. Specifically, the polymer may include one or more selected from 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), polyetheretherketone (PEEK), polyethersulfone (PES), polyphenylene oxide (PPO), and polyphenylene sulfide (PPS), but is not limited thereto. The above metal may include one or more selected from stainless steel, titanium, nickel, tantalum, zirconium, and alloys thereof, but is not limited thereto. The above ceramic may include one or more selected from fused silica, alumina, zirconia, silicon carbide, silicon nitride, boron nitride, and titanium diboride, but is not limited thereto.

[0046] In one embodiment, the core substrate may have one or more shapes selected from a one-dimensional shape, a two-dimensional shape, and a three-dimensional shape.

[0047] In one embodiment, the one-dimensional shape may include one or more selected from staple fibers, filament fibers, and rods. 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 a length sufficient for biological polymer synthesis to occur, and the composite solid support may be loaded into a reactor in an entangled form to be utilized in the biological polymer synthesis reaction.

[0048] In one embodiment, the two-dimensional shape may include one or more selected from spunbond non-woven fabric, meltblown non-woven fabric, needle-punched non-woven fabric, hydroentangled non-woven fabric, woven fabric, knitted fabric, porous membrane, polymeric film, and mesh. As a specific example, the two-dimensional shape of the core substrate may include a form in which a plurality of core substrates of the one-dimensional shape are randomly arranged. Additionally, the two-dimensional shape may exhibit random porosity by forming pores between the plurality of cores of the one-dimensional shape through the random arrangement.

[0049] In one embodiment, the three-dimensional shape may include an open cell foam, a macropored sphere, or both.

[0050] In one embodiment, the length of one cross-section of the core substrate is 1 mm or more, and the functional coating may have a size sufficient to participate in the synthesis of the intended biological polymer. As a specific example, if the core substrate has a one-dimensional shape, the core substrate can be implemented to act as a support for a composite solid support through entanglement, such as a fiber. In addition, if the core substrate has a two-dimensional or three-dimensional shape, it can act as a support itself, such as a nonwoven fabric (two-dimensional) or 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 of infinite length can also serve as a support.

[0051] In one embodiment, the functional coating may comprise a polymer of one or more main monomers and an active monomer. As a specific example, the functional coating may comprise a polymer of a first monomer and the active monomer, or a polymer of a first monomer, a second monomer, and the active monomer. Additionally, the functional coating of the present invention may be a polymer formed through a polymerization reaction between one or more main monomers and an active monomer that provides reaction sites as main components.

[0052] In one embodiment, the first monomer may comprise one or more selected from bisacrylamide-based crosslinking agents, triazines substituted with methacryloyl groups and alkenyl groups, and acrylate-based crosslinking agents; specifically, the first monomer may comprise polyethylene glycol-based, for example, polyethylene glycol diacrylate. Specifically, the first monomer may be used without limitation as long as it is a material capable of reacting with or bonding with an active monomer for exposing functional groups to the interior and / or surface of a functional coating. As a non-limiting example, the first monomer may comprise one or more 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 comprise 2-hydroxyethyl acrylate, It may include one or more 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).

[0053] In one embodiment, the active monomer is intended to provide a functional group inside and / or on the surface of the functional coating and may include one or more selected from acrylate-based, acrylamide-based, and methacrylamide-based materials. As a non-limiting example, the active monomer is 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 It may contain one or more selected from N-(2-aminoethyl)acrylamide hydrochloride.

[0054] In one embodiment, the composite solid support may comprise 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 from a polymer of one or more main monomers and the active monomer. Additionally, in the heterogeneous composite solid support, the functional coating comprising a pulverized conventional solid support may be in the form of being coated on the core substrate. The above-mentioned conventional solid support may comprise one or more selected from polystyrene / divinylbenzene copolymer (PS / DVB), crosslinked polyethyleneglycol polymer, poly-ε-lysine / sebacic acid copolymer, controlled pore glass, amino-polyacrylamide-resin fiber, cellulose, and hydroxylated polypropylene (hydroxylated PP), but is not limited thereto.

[0055] In one embodiment, the one-dimensional core substrate may have a diameter (cross-section) of 10 μm to 100 μm. Specifically, it may be 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, but is not limited thereto. If the diameter of the above-mentioned one-dimensional core substrate is less than 10 μm, it becomes too thin, causing the core substrate to clump together and resulting in a decrease in mechanical properties; if it exceeds 100 μm, it may result in a problem where the loading density of the functional coating becomes excessively low.

[0056] 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, and 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, 300 μm to It may be 0.5 mm, 350 μm to 0.5 mm, or 400 μm to 0.5 mm, but is not limited thereto. 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 to the interior of the functional coating.

[0057] In one embodiment, the porosity of the core substrate may be 50% to 95%. If the core substrate is one-dimensional, the core substrate may form pores through entanglement, such as fibers, and if the core substrate is two-dimensional or three-dimensional, it may contain pores inside, such as nonwoven fabrics or foams. In cases where pores are present, the porosity of the core substrate may be 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%, but is not limited thereto. Here, there are no specific limitations on the pore size of the porous membrane substrate, and if the porosity is less than 50%, the pores may become clogged even after the functional coating has swollen, preventing biological polymer synthesis from occurring within the functional coating. Additionally, if the porosity exceeds 95%, it may be difficult to maintain mechanical properties.

[0058] In one embodiment, the thickness of the functional coating may be 0.1 μm to 1000 μm. The thickness of the functional coating may be selected as an appropriate thickness depending on the type of biological polymer desired and the one-dimensional, two-dimensional, or three-dimensional shape of the core substrate. 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 μm. It may be possible, but is not limited thereto. If the thickness of the functional coating is excessively small, such as less than 0.1 μm, a problem arises in which the loading density of the functional coating becomes very low.In addition, if the thickness of the functional coating is excessively large, such as when it exceeds 1000 μm, the functional coating itself is manufactured through a non-uniform polymerization reaction, and a problem of reduced purity may occur during the biological polymer synthesis process.

[0059] The above composite solid support may not be a shell-core structure or a bead structure. Additionally, the above solid support may not include a core in a shell-core structure.

[0060] 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 hydrophilically treated surface.

[0061] In one embodiment, the functional coating may additionally include a linker.

[0062] In one embodiment, the linker may include one or more 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 an HMBA (TFA stable) linker.

[0063] In one embodiment, the functional coating has a functional group exposed internally and / or on its surface, and the linker may be bonded to the functional group. A biological polymer comprising a plurality of monomer units may be synthesized by adding an additional monomer unit (amino acid residue) to a monomer unit (amino acid residue) connected through the functional coating (functional group)-linker.

[0064] In one embodiment, the biological polymer may comprise one or more selected from peptides, oligonucleotides, and peptide nucleic acids (PNA), but is not limited thereto.

[0065] FIG. 1 is a drawing illustrating a biological polymer synthesis system according to one embodiment of the present invention.

[0066] 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 containing at least one of an amino acid, a reagent, and an additive.

[0067] In the following, the ‘amino acid mixture’ is a liquid containing reagent(s) as well as amino acids as used herein, wherein the ‘amino acid’ may be modified or unmodified and may optionally be pre-activated, and the amino acid mixture may also contain peptides.

[0068] In the following, '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).

[0069] In the following, 'reagent' is used to mean coupling reagent, deprotection reagent, additive, base, and other reagents used in synthesis.

[0070] In the following, '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 various types of single substances or mixed substances depending on the process.

[0071] In one embodiment, a biological polymer synthesis system (1) may be subjected to a flow-through process for solid phase peptide synthesis (SPPS) that enables the sequential synthesis of at least one amino acid.

[0072] In one embodiment, the biological polymer synthesis system (1) may include a mixing reservoir (100), a pump (200), a temperature controller (300), and a reactor (400). Additionally, the biological polymer synthesis system (1) may further include a valve (500), a solvent reservoir (500), a sensor (SE), and a detector (DE).

[0073] The main line (ML) can provide a circulation piping line by connecting a mixing storage tank (100), a pump (200), a temperature controller (300), and a reactor (400). Additionally, a valve (500) is installed in the main line (ML), and the valve (500) can be connected to a first branch line (SL1) and a second branch line (SL2).

[0074] The mixing storage tank (100) can provide a space for a fluid containing at least one of an amino acid, a reagent, and an additive. The mixing storage tank (100) has an opening / closing valve (not shown) placed at the outlet end to control the type and flow rate of the fluid flowing into the main line (ML).

[0075] The mixing tank (100) may mix amino acids, reagents, and additives 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 adjusting the opening amount of each chamber. As an example, in each synthesis step of the biological polymer of the SPPS method described below, the corresponding fluid may be introduced into the mixing tank, mixed and held, and then introduced into the reactor.

[0076] 1) Deprotection step: Deprotection reagent (piperidine)

[0077] 2) Washing step: Solvent (DMF)

[0078] 3) Coupling step: Amino acid, coupling reagent (DIC), additive (Oxyma pure)

[0079] 4) Washing step: Solvent (DMF)

[0080] 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 to cause the fluid to flow from the mixing storage tank (100) to the reactor (400). In FIG. 1, the pump (200) is located between the mixing storage tank (100) and the reactor (400) in terms of fluid flow, but since the fluid forms a circulating flow, it is not necessary for the pump (200) to be located between the mixing storage tank (100) and the reactor (400).

[0081] The pump (200) can be any type of device that provides suction and discharge forces to the fluid. For example, the pump (200) can be a gear pump, screw pump, vane pump, cap pump, piston pump, plunger pump, diaphragm pump, centrifugal pump, etc.

[0082] Additionally, the pump (200) can be any type of pump, such as a mechanical displacement type micropump and an electromagnetic motion type micropump. A mechanical displacement type micropump is a pump that utilizes the motion of a solid or fluid, such as a gear or diaphragm, to create a pressure difference to induce fluid flow, and includes diaphragm displacement pumps, fluid displacement pumps, rotary pumps, etc. An electromagnetic motion type micropump is a pump that utilizes energy in the form of electricity or magnetism directly for fluid movement, and includes electrohydrodynamic pumps (EHD), electroosmotic pumps, magnetohydrodynamic pumps, electrowetting pumps, etc.

[0083] 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 positioned downstream of the mixing storage tank (100) and upstream of the reactor (400) to cool or heat the fluid containing one or more selected from amino acids, reagents, and additives. In FIG. 1, the temperature controller (300) is located between the mixing storage tank (100) and the reactor (400) in terms of fluid flow, but since the fluid forms a circulating flow, it is not necessary for it to be located between the mixing storage tank (100) and the reactor (400).

[0084] The temperature controller (300) may be various devices that transfer heat to a fluid through heat exchange or receive heat from a fluid. For example, the temperature controller (300) may be an electric heating device, an induction heater, or a microwave cavity.

[0085] 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 from 25°C to 100°C. Additionally, the temperature controller (300) can control the temperature of the fluid to 50°C to 80°C.

[0086] Figure 2 is a drawing illustrating the reactor of Figure 1.

[0087] Referring to FIGS. 1 and 2, the reactor (400) can circulate fluid discharged from the mixing storage tank (100).

[0088] The reactor (400) may be equipped with a column (410) and a composite solid support (420).

[0089] The column (410) may have an inlet (411) for fluid inflow and an outlet (412) for fluid outflow. The column (410) may have an internal space, and a composite solid support (420) may be placed therein.

[0090] The column (410) can have various sizes depending on the type of biological polymer being synthesized, the amount of biological polymer synthesized, the flow rate of the fluid, the velocity of the fluid, and the type of composite solid support.

[0091] The inlet (411) and outlet (412) of the column (410) are positioned on the main line (ML), so that fluid entering through 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 a composite solid support (420) positioned in the internal space of the column (410).

[0092] The composite solid support (420) can be placed inside the column (410). The solid support (420) can be filled inside the column (410).

[0093] The composite solid support (420) can be placed in various positions and shapes within the internal space of the column (410). The composite solid support (420) can be filled in a predetermined position within the internal space of the column (410).

[0094] For example, a composite solid support (420) can be loaded into the internal space of the column (410).

[0095] For example, if the core substrate of the composite solid support (420) is in the form of entangled filaments (FM), fluid can pass through between the filaments (FM). The fluid flow path may vary depending on the method of loading the composite solid support (420). For example, if a composite solid support in the form of a spirally rolled shape is loaded, fluid can flow between each layer of the spiral composite solid support and in the porous space created by the entanglement.

[0096] The composite solid support (420) may have a core substrate (BM) and a functional coating (CO).

[0097] As an example, the core substrate (BM) may have multiple filaments intertwined to form random porosity. The functional coating (CO) may be a polymer for biological polymer synthesis coated on the core substrate so that functional groups such as amine groups and carboxyl groups are exposed.

[0098] 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.

[0099] The core substrate (BM) may possess solvent resistance, thermal resistance, or both of the above characteristics. Unlike the functional coating (CO), the core substrate (BM) does not have swelling or dissolving properties in solvents, and is self-standing and can serve as a support for a composite solid support. In addition, the core substrate (FM) is thermally resistant and can serve as a support without being altered even if the synthesis of the biological polymer proceeds at high temperatures.

[0100] A functional coating can surround the core substrate (FM) and provide a region where biological polymers are synthesized.

[0101] In one embodiment, the functional coating (CO) may have the characteristic of swelling in a solvent.

[0102] The functional coating (CO) may contain functional groups on its surface and inside. Specifically, the functional coating contains functional groups, and due to its swelling characteristic in a solvent, it facilitates the penetration of reactants into the functional coating due to a concentration gradient. As a result, the functional groups on the surface and inside the functional coating are easily exposed to the reactants, allowing for high synthesis efficiency even at low loading densities.

[0103] The valve (500) is positioned between the reactor (400) and the storage tank (100) to establish a path for the discharge of waste. The valve (500) can control the direction of fluid flow so that waste generated after the reaction in the reactor (400) is discharged to the second branch line (SL2). As an example, the valve (500) may be a 4-channel valve, and the 4-channel valve may be connected to the mixing storage tank (100), the reactor (400), the solvent storage tank (600), and the second branch line (SL2).

[0104] The valve (500) can control the circulation of fluid and the discharge of waste. The valve (500) can increase the synthesis rate of the biological polymer by setting the circulation flow of the reactant fluid during each stage of SPPS synthesis, and can allow the remaining material from each stage to be discharged to the second branch line (SL2) to proceed to the next stage.

[0105] The solvent storage tank (600) can store solvents used for biological polymer synthesis and cleaning. The solvents can be selected in various ways depending on the synthesis process.

[0106] A solvent storage tank (600) may be placed on a first branch line (SL1). The solvent storage tank (600) is connected to the first branch line (SL1), so that, depending on each step of SPPS synthesis, the solvent may move to a valve (500) or to a mixing storage tank (100).

[0107] Before proceeding with the wash process during the synthesis process, any material remaining in the piping from the previous step must be removed. At this time, the solvent discharged from the solvent storage tank (600) is discharged into the first branch line (SL1) so that the main line (ML) can be washed.

[0108] The sensor (SE) is positioned on the main line (ML) and can sense at least one of the temperature, pressure, and flow rate of the fluid flowing along the main line (ML).

[0109] A detector (DE) is placed on the main line (ML) to detect the progress of biological polymer synthesis. As an example, the detector may be placed at least one of the inlet and outlet of the reactor (400) to detect the circulating fluid. In FIG. 1, the detector (DE) is located between the reactor (400) and the mixing storage tank (100) in the fluid flow, but since the fluid forms a circulating flow, it is not necessary for the detector (DE) to be located between the reactor (400) and the mixing storage tank (100).

[0110] 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 fluid circulation path.

[0111] The first branch line (SL1) branches off 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 solvent from the solvent storage tank (600) to the main line (ML), thereby removing residual material in the main line (ML).

[0112] The second branch line (SL2) can be branched from the valve (500) to discharge waste.

[0113] Preferred embodiments are presented below to aid in understanding the present invention. However, the following embodiments are provided merely to facilitate a better understanding of the invention, and the scope of the invention is not limited by the following embodiments.

[0115] Example 1. Analysis of pressure drop in a reactor according to flow rate

[0116] Figure 3 is a graph showing the pressure drop according to the flow rate to compare the reactor of the present invention with a comparative example.

[0117] (Test equipment)

[0118] - Column (Intertec empty cartridge column): 27 mL, ID / Hight: 12.8*60 mm

[0119] - Pump: LEAD FLUID CT3001, Pump Head: Fluid-o-Tech / MG209

[0120] - Reagents used: DMF (Dimethylformaminde, SAMJHUN), Rink-Amide-MBHA-Resin (GL Biochme, 0.5mmole / g)

[0121] - Pressure measuring equipment: SMC PSE560-C01 pressure sensor, SMC PSE200A digital pressure sensor controller

[0122] For the example, 5.0 g (0.5 mmol, loading density: 0.1 mmol / g) of a composite solid support was placed in a column, and for the comparative example, 1.0 g (0.5 mmol, loading density: 0.5 mmol / g) of a particulate solid support (PS / DVB) was placed in a column, then 20 mL of DMF solvent was added and the composite solid support was swollen for 30 minutes.

[0123] Subsequently, 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.

[0124] DMF was flowed at a flow rate of 40 mL / min (1.48 CV / min) through an empty column, a composite solid support column (Example), and a particulate solid support column (Comparative Example), respectively, and the pressure at the inlet and outlet was measured. Subsequently, the pressure at the inlet and outlet of the column was measured while changing the flow rate to 50 ml / min (1.85 CV / min), 100 ml / min (3.7 CV / min), 200 ml / min (7.4 CV / min), 300 ml / min (11.11 CV / min), 400 ml / min (14.81 CV / min), and 500 ml / min (18.52 CV / min).

[0125] The pressure drop values ​​at the inlet and outlet of the empty column were set as the reference value (Zero). The additional pressure drop values ​​were calculated by comparing the inlet and outlet of the column containing the composite solid support with the reference value, and the additional pressure drop values ​​were calculated by comparing the inlet and outlet of the column containing the particulate solid support with the reference value.

[0126] Flow rate (ml / min) Comparative example Examples 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

[0127] Referring to Figure 3 and Table 1, the composite solid support of the present invention exhibited a small pressure drop even as the flow rate increased. This is because fluid can pass through the empty spaces of the composite solid support, resulting in a small pressure drop and increased fluid flowability even as the flow rate increases. In contrast, the conventional particulate solid support (bead-type solid-phase synthetic resin polymer (PS / DVB)) exhibited a large pressure drop as the flow rate increased, and fluid was not discharged at 300 ml / min. This is because the conventional particulate solid support clumps together to fill the space within the column when the flow rate increases, reducing the space available for fluid passage; consequently, the pressure measured at the outlet drops significantly, and fluid flowability decreases.

[0129] Example 2. Analysis of pressure drop of composite solid support according to column size

[0130] Figure 4 is a graph showing the pressure drop according to the size of the reactor of the present invention.

[0131] (Test equipment)

[0132] - A column (Intertec empty cartridge column): 27 mL, ID / Hight: 12.8*60 mm

[0133] - B column (Intertec empty cartridge column): 108 mL, ID / Hight: 21.4*76 mm)

[0134] - C column (Intertec empty cartridge column): 385mL, ID / Hight: 26.8*127 mm

[0135] - Pump: LEAD FLUID CT3001, Pump Head: Fluid-o-Tech / MG209

[0136] - Reagents used: DMF (Dimethylformaminde, SAMJHUN), Rink-Amide-MBHA-Resin (GL Biochme, 0.5mmole / g)

[0137] - Pressure measuring equipment: SMC PSE560-C01 pressure sensor, SMC PSE200A digital pressure sensor controller

[0138] 5 g of the composite solid support was loaded onto column A, 20 ml of DMF solvent was added, and it was swollen for 30 minutes. 20 g of the composite solid support was loaded onto column B, 80 ml of DMF solvent was added, and it was swollen for 30 minutes. 70 g of the composite solid support was loaded onto column C, 300 ml of DMF solvent was added, and it was swollen for 30 minutes.

[0139] The discharge tubes of the pumps were connected to each column, pressure sensors were installed at the inlet and outlet of each column, and each pressure sensor was connected to a controller.

[0140] DMF was flowed through columns A, B, and C at a flow rate of 0.5 column volume (mL) / min, respectively, and the pressures at the inlets and outlets were measured. Subsequently, the flow rates were changed to 1 CV / min, 2 CV / min, 3 CV / min, 4 CV / min, and 5 CV / min, and the pressures at the inlets and outlets of the columns were measured.

[0141] The pressure drop values ​​generated at the inlet and outlet of each empty column were set as the reference value (Zero). The additional pressure drop values ​​generated at the inlet and outlet of columns A, B, and C, which contained the composite solid support, were calculated by comparing them to the reference value.

[0142] Flow velocity (CV / 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

[0143] Referring to Figure 4 and Table 2, the composite solid support of the present invention exhibited a small pressure drop in the fluid at various flow rates, even when the column volume was varied. This is because the fluid can pass through the empty spaces of the composite solid support, so the pressure drop is small and the fluid flowability increases even when the flow rate increases. Accordingly, the composite solid support of the present invention consistently exhibits a small degree of pressure drop regardless of the column volume.

[0145] Example 3. Analysis of pressure drop of a composite solid support according to temperature

[0146] Figures 5 and 6 are graphs showing the pressure drop according to temperature of the reactor of the present invention and a comparative example.

[0147] When the flow velocity is the same, a higher rotational speed of the same pump results in greater back pressure. Based on this, the rotational speed of the pump at a specific flow rate was measured to compare the back pressure applied to each solid support.

[0148] A 27 ml column loaded with the composite solid support (0.5 mmol) used in the present invention (Example), an empty column (Comparative Example 1), and a 27 ml column loaded with a conventional particulate solid support (Rink-Amide-MBHA-Resin, 0.5 mmol, 1.25 g) (Comparative Example 2) were compared.

[0149] As shown in Table 3 and Figure 5, the RPM values ​​of the pump were recorded when using DMF solvent at 25℃ at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 ml / min.

[0150] When DMF was used at 25℃, the column loaded with a composite solid support (Example) did not differ significantly in pump rotation speed from the empty column (Comparative Example 1). On the other hand, the column loaded with a particulate solid support (Comparative Example 2) stopped operating due to high back pressure at 250 ml / min.

[0151] Flow rate (ml / min) Examples 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

[0152] As shown in Table 4 and Figure 6, the RPM values ​​of the pump were recorded when using DMF solvent at 50°C at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 ml / min. When DMF was used at 50°C, the back pressure decreased as the viscosity of the solution decreased. However, for the particulate solid support (Comparative Example 2), the pump stopped operating due to high back pressure at 350 ml / min.

[0153] Flow rate (ml / min) Examples 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 -

[0154] As shown in Table 5, the RPM values ​​of the pump were recorded when using DMF solvent at 70°C at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 ml / min. When DMF was used at 70°C, the back pressure decreased as the viscosity of the solution decreased. However, for the particulate solid support (Comparative Example 2), the pump stopped operating due to high back pressure at 350 ml / min.

[0155] Flow rate (ml / min) Examples 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 -

[0156] In addition, the rotational speed of the pump was measured at room temperature using Piperidine 20% in DMF (0.1M Oxymapure), which is widely used in solid-phase peptide synthesis reactions. Even in this case, the particulate solid support stopped operating at 250 ml / min due to high back pressure. In other words, it was confirmed that the composite solid support of the present invention experiences less back pressure than conventional particulate solid supports. Considering that the volume and weight of the composite solid support increase as longer peptides are synthesized, thereby increasing back pressure, the composite solid support of the present invention can improve the synthesis performance of a flow reactor by enhancing fluid flowability, unlike conventional particulate solid supports.

[0157] 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 the synthesis reaction.

[0158] Conventional particulate solid-phase synthesis resin polymers clump together at high flow rates and fill the internal space of the column without any empty space, which reduces the flowability of the reactor. Consequently, a high pressure drop occurs at the outlet of the reactor, resulting in high back pressure. High back pressure prevents the synthesis scale of the synthesis system from being significantly increased.

[0159] A flow reactor and a biological polymer synthesis system including the same according to one embodiment of the present invention have increased flowability of the reactor by means of a composite solid support. Thus, the reactor experiences a low pressure drop and consequently achieves low back pressure, thereby enabling an increase in the synthesis scale.

[0160] Specifically, under the same pressure conditions, lower back pressure reduces flow rate loss, thereby promoting mixing and diffusion between the reactants and the reaction solution within the column. Biological polymer synthesis systems exhibit improved flow dynamics with increasing flow rates, leading to enhanced reaction efficiency and yield.

[0161] Furthermore, reducing back pressure decreases wear and mechanical stress on internal reactor components, thereby improving the durability of the entire system. Additionally, since pumps providing various pressures can be applied, versatility and the range of options can be increased. In particular, high reaction efficiency and yield can be achieved even when using low-pressure pumps.

[0162] In addition, reducing back pressure lowers the risk of damage to fluid-carrying pipes and connections, and by reducing fluid leakage, it can increase the durability of the system as well as the stability and reliability of the process.

[0163] In addition, since reducing back pressure allows for precise control of the fluid passing through the reactor, the reproducibility of the reaction process is improved, and biological polymers of consistent quality and high quality can be synthesized.

[0165] Example 4. Preparation of a composite solid support

[0166] (1) Preparation of polymers for functional coatings

[0167] Experiments were conducted to measure the synthesis performance of biological polymers using the previously identified flow reactor. First, a polymer for functional coatings used on composite solid supports was prepared. Each reagent listed 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 only need to satisfy the respective mass ratio (parts by weight) ranges, and the solvent can be used regardless as it is scheduled to be dried later. In the experiment, the total weight of each reagent and solvent (DW) was measured to be 100 g [the solvent may be used within 10 wt% to 90 wt% of the total (reagent + solvent)].

[0168] 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 surfactants Sodium Dodecyl Sulfate 0~25

[0169] Subsequently, 0.5 g of 2-hydroxy-2-methylpropiophenone, a curing reaction initiator, was added to the above solution and uniformly mixed to prepare a functional coating solution. The functional coating solution was uniformly applied to a Petri dish and irradiated with ultraviolet energy ranging from 10 mJ / cm² to 1,000 mJ / cm² sufficiently to cure it. After curing was complete, reaction residues were removed using ethanol to obtain a functional coating polymer disc. The mass of the dried functional coating polymer was measured to be 45.7 g. The prepared functional coating polymer was rapidly frozen using liquid nitrogen and then ground using a mortar and pestle. Subsequently, functional coating polymer particles with a particle size range of 100-200 mesh (74-149 μm) were obtained using 100- and 200 mesh sieves.

[0171] (2) Preparation of a composite solid support for biological polymer synthesis

[0172] To manufacture a composite solid support, the functional coating solution (before curing) from (1) of Example 4 above and a core substrate (polypropylene, spunbond nonwoven fabric, 40 g / m²) were used. 2 20 g was prepared. The core substrate was sufficiently immersed in the functional coating solution, and after removing the core substrate from the functional coating solution, it was subjected to UV curing at 10 mJ / cm² in a UV curing chamber. 2 - 1,000 mJ / cm 2 The ultraviolet energy was sufficiently irradiated.

[0173] After the curing process was completed, the coated core substrate was subjected to removal of reaction residues using ethanol and drying. A composite solid support coated with a functional coating polymer on a core substrate was obtained (solid support 30.4 g, 60.8 g / m² 2 ...and 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.

[0175] Example 5. Synthesis of biological polymer using a composite solid support

[0176] Compared to the method of a batch-type automatic synthesizer (CEM Liberty, USA), we investigated whether peptide synthesis is possible and whether there is a difference in the purity and yield of the synthesized peptide when synthesizing using the SPPS method with a biological polymer synthesis system including a column-type flow reactor containing a solid support for biological polymer synthesis according to one embodiment.

[0178] (1) Synthesis of peptides using a batch-type automated synthesizer

[0179] After adding 0.1 mmol each of a conventional solid-phase synthesis particle-type solid support and a composite solid support according to one embodiment to an automatic synthesizer, the synthesis reaction of the ACP (acyl carrier protein) model peptide (65-74) of SEQ ID NO. 1 was carried out under the same conditions as shown in Table 7 below.

[0180] Sequence No. 1: VQAAIDYING

[0181] Reaction step Reagent (Solvent: DMF) Volume (mL) Temperature (°C) Hours (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) - -

[0182] After the peptide synthesis reaction was completed, the particulate solid support and the composite solid support were each thoroughly washed with ethanol and dried. Subsequently, a cleavage reaction was carried out for 2 hours using a cleavage solution (95% TFA, 2.5% TSI, 2.5% DW). Afterward, the samples were precipitated using cold ether and centrifuged to obtain a solid. The solid was then washed twice more with 15 ml of cold ether and thoroughly dried using vacuum 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.

[0183] Types of solid supports water(%) transference number(%) Composite solid support 85.1 88.4 Particulate solid support 81.4 71.5

[0184] In the analysis of peptide synthesis of a particulate solid support and a biological polymer composite solid support according to one embodiment using a conventional commercial batch-type SPPS synthesizer (Liberty, CEM, USA), the yield and purity of ACP peptides were improved in the composite solid support according to one embodiment compared to the particulate solid support.

[0186] (2) Synthesis of peptides using a flow-based column synthesizer

[0187] To compare the synthesis performance of particulate solid support and homogeneous composite solid support in a flow reactor, 0.5 mmol each of particulate solid support and composite solid support were added to a 27 mL column, and the reaction steps were carried out while changing the type of amino acid under the conditions shown in Table 9 below to synthesize the ACP (acyl carrier protein) model peptide (65-74) of SEQ ID NO. 1. Afterward, the dry mass was measured to calculate the yield, and the purity of the synthesized peptide was measured using high-performance liquid chromatography (HPLC).

[0188] Reaction step Reagent (Solvent: DMF) Volume (mL) Temperature (°C) Hours (minutes) Flow rate (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 - -

[0189] Synthesis performance in a flow reactor using a composite solid support was found to be 95.8% in purity and 93.5% in yield, demonstrating superior performance in both purity and yield compared to a batch reactor. In contrast, when ACP (65-74) peptide synthesis was carried out by filling a flow reactor with a particulate solid support, a backpressure issue occurred during synthesis, preventing proper synthesis. This occurs because the particulate solid support clogs the column of the flow reactor due to swelling. The composite solid support, which exists with a functional coating on the core substrate, does not have the potential for such problems to occur, indicating that it is more suitable for use in flow reactors. Through this, it was confirmed that the composite solid support is suitable for both batch and flow synthesis systems compared to the existing particulate solid support, and particularly when applied to a flow synthesis system, the yield and purity of ACP peptide were over 90%, demonstrating significantly superior efficacy in the synthesis of biological polymers.

[0191] Example 6. Analysis and Evaluation of Changes in Yield and Purity of Synthesized Peptides According to Flow Rate of Biological Polymers in a Flow Reactor

[0192] The yield and purity of the synthesized peptide according to the flow rate were verified using a composite solid support as in Example 5. ACP was synthesized in the same manner 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.

[0193] Flow rate (mL / min) transference number(%) water(%) 100 84.9 85.3 200 88.3 90.5 300 91.1 95.7

[0194] Figures 9 to 11 show HPLC graphs of ACP synthesized according to flow rate using a composite solid support for biological polymer synthesis according to one embodiment. When the solid support for biological polymer synthesis according to one embodiment was used, it was confirmed that the purity and yield of the peptide increased when the flow rate of the column synthesizer was increased.

[0195] The above results indicate that a flow-based reactor including a column is a more suitable reactor for a composite solid support according to one embodiment.

[0196] As such, the present invention has been described with reference to the embodiments illustrated in the drawings, but this is merely illustrative, and those skilled in the art will understand that various modifications and equivalent alternative embodiments are possible therefrom. Accordingly, the true technical scope of protection of the present invention should be determined by the technical spirit of the appended claims. Explanation of the symbols

[0197] 1: Biological polymer synthesis system 100: Mixed storage tank 200: Pump 300: Thermostat 400: Reactor 500: Valve 600: Solvent storage tank

Claims

Claim 1 A biological polymer synthesis system comprising: a mixing tank in which an amino acid and at least one of a reagent and an additive are mixed; a reactor having a column into which a fluid discharged from the mixing tank is introduced and which has an inlet for the fluid to be introduced and an outlet for the fluid to be discharged, and a composite solid support disposed inside the column; a temperature controller disposed between the mixing tank and the reactor to control the temperature of the fluid introduced into the reactor; and a pump providing a driving force to cause the fluid to flow from the mixing tank to the reactor; wherein the composite solid support comprises a core substrate; and a functional coating located on the core substrate. Claim 2 delete Claim 3 In claim 2, the composite solid support is a biological polymer synthesis system that is self-standing when the fluid passes through the column. Claim 4 In claim 2, the composite solid support is a biological polymer synthesis system in which the loading density of the functional coating is 0.01 mmol / g to 2 mmol / g. Claim 5 A biological polymer synthesis system according to claim 1, wherein, when the fluid passes through the reactor, the pressure drop between the inlet and the outlet is 10 bar or less at a flow rate of 40 CV / min or less. Claim 6 A biological polymer synthesis system according to claim 1, wherein the temperature of the fluid flowing into the reactor after passing through the temperature controller is 120°C or lower. Claim 7 A biological polymer synthesis system according to claim 1, further comprising a valve disposed between the reactor and the mixing storage tank for discharging waste. Claim 8 A biological polymer synthesis system according to claim 1, further comprising: a main line connecting the mixing storage tank, the pump, the temperature controller, and the reactor; and a first branch line branched from the main line between the mixing storage tank and the reactor and connected to the mixing storage tank. Claim 9 A biological polymer synthesis system according to claim 8, wherein the fluid has a circulating flow through the main line. Claim 10 A biological polymer synthesis system according to claim 8, further comprising a solvent storage tank disposed on the first branch line. Claim 11 A biological polymer synthesis system according to claim 10, wherein the solvent storage tank is connected to the valve and the mixing storage tank via the first branch line. Claim 12 A biological polymer synthesis system according to claim 8, further comprising a sensor disposed on the main line and sensing at least one of the temperature, pressure, and flow rate of the fluid flowing along the main line. Claim 13 A biological polymer synthesis system according to claim 8, further comprising a detector disposed on the main line to detect the progress of the synthesis of the biological polymer. Claim 14 A biological polymer synthesis system according to claim 7, further comprising a second branch line branched from the valve to discharge the waste.

Images