Flow reactor and biopolymer synthesis system comprising same

Abstract

The present invention relates to a flow reactor and a biopolymer synthesis system comprising the same, the biopolymer synthesis system comprising: a mixing storage tank for mixing at least one of an amino acid, a reagent and an additive; a reactor into which the fluid discharged from the mixed storage tank flows; a temperature regulator which is provided between the mixing storage tank and the reactor, and which regulates the temperature of the fluid flowing into the reactor; and a pump for providing a driving force to cause the fluid to flow from the mixing storage tank to the reactor, the reactor comprising: a cylinder having an inlet through which the fluid flows in and an outlet through which the fluid is discharged; and the composite solid supporting body is arranged in the column body.

Description

Technical Field

[0001] This invention relates to a flow reactor and a biopolymer synthesis system. Background Technology

[0002] Peptide pharmaceuticals generate billions of dollars in sales annually in the diabetes, obesity, and oncology industries and are expanding into new drug development for emerging diseases such as cardiovascular and neurodegenerative diseases. Currently, the short in vivo half-life of peptides is being addressed by introducing non-natural amino acids. Several peptide pharmaceuticals with dosing cycles exceeding one week are already on the market, and multiple oral administration candidates are undergoing clinical trials with the U.S. Food and Drug Administration (FDA). Consequently, the demand for peptide pharmaceutical production is increasing by nearly 10% annually. To meet this demand, a new synthetic platform is urgently needed to address both increased productivity and environmental, social, and governance (ESG) concerns (such as compliance requirements related to N,N-dimethylformamide (DMF) used in traditional chemical synthesis methods).

[0003] Peptide pharmaceuticals are mainly prepared through biological or chemical methods. Biological methods using genetic recombination or transgenic plants and animals have two major drawbacks: firstly, it is difficult to produce peptide pharmaceuticals with increased half-life due to the use of non-natural amino acids (the current industry trend) or PEGylation; secondly, it is difficult to remove various endogenous contaminants from organisms or exogenous contaminants generated during the process.

[0004] In chemical methods, liquid-phase reaction processes are low-cost, but they can only synthesize peptides composed of fewer than 10 amino acids, thus preventing the production of peptide pharmaceuticals composed of more than 20 amino acids. In the preparation of PEGylation peptide pharmaceuticals composed of 20–50 different types of natural / non-natural amino acids, the most suitable method is solid-phase peptide synthesis (SPPS).

[0005] To improve the production capacity of various biopolymers, including peptides, various types of reactors are being researched in the SPPS (Synthetic Processing System) field. Typical reactors include batch reactors and flow reactors. For large-scale synthesis, the scale of batch reactors has gradually increased from 10-50 mL to the current 100-500 L units, resulting in various improvements in solution delivery, stirring, pressure regulation, and filtration methods. At the laboratory scale, recently introduced flow reactors have demonstrated the ability to achieve rapid, high-purity, and high-yield peptide synthesis.

[0006] However, batch reactors require adjustments to reactor size based on reaction scale, which presents several challenges, including: temperature transfer between the center and surface of the reaction mixture, differences in time required for homogeneous mixing, low reproducibility of reaction temperature / mixing, and difficulties in yield and quality management due to reproducibility issues.

[0007] In contrast, conventional flow reactors increase the reaction surface area by passing through a tubular reactor and mixing at high speed, thus offering the following advantages: achieving uniform temperature transfer and inducing chemical reactions, resulting in significant reaction reproducibility, which in turn facilitates quality and yield management.

[0008] Current flow reactors offer the advantage of rapid synthesis reactions, but when using 200 mg of PS / DVB, they suffer from back pressure of 30-50 bar. Such high back pressure clearly indicates that the synthesis scale cannot be significantly increased. Furthermore, the flow reactor developed by Vaportek in the UK uses an adjustable syringe, solving the high back pressure drawback of packed beaded resins, but it has the following limitations: the usable flow rate is very small, and the pressure regulation characteristic of the syringe makes it difficult to significantly increase the synthesis scale.

[0009] In addition, existing granular resins have the limitations of granular structure, cannot be self-standing, and move and pack through flow, thus having the structural limitation of increased back pressure in flow reactors.

[0010] Therefore, a new solid support material suitable for SPPS flow reactors and capable of self-supporting, as well as a biopolymer synthesis system using it, has been developed. Summary of the Invention

[0011] The problem the invention aims to solve The purpose of this invention is to provide a SPPS flow reactor with improved synthesis efficiency and yield, as well as high durability and stability, and a biopolymer synthesis system including the same.

[0012] means for solving problems An embodiment of the present invention provides a biopolymer synthesis system, the biopolymer synthesis system comprising: a mixing storage tank for mixing at least one of amino acids, reagents, and additives; a reactor into which fluid discharged from the mixing storage tank flows; a temperature regulator disposed between the mixing storage tank and the reactor and regulating the temperature of the fluid flowing into the reactor; and a pump for providing driving force to cause the fluid to flow from the mixing storage tank to the reactor, the reactor comprising: a column having an inlet for the fluid inflow and an outlet for the fluid outflow; and a composite solid support disposed inside the column.

[0013] Invention Effects The flow reactor and biopolymer synthesis system comprising the present invention provide a biopolymer synthesis process with improved efficiency and yield. Specifically, the biopolymer synthesis system is characterized by the use of a flow reactor loaded with a composite solid support, exhibiting low pressure drop even under high flow rate conditions, and maintaining excellent efficiency and yield across a variety of fluid temperature ranges.

[0014] In addition, the flow reactor and biopolymer synthesis system including the present invention can improve the overall durability and stability of the equipment and can precisely control the process. Attached Figure Description

[0015] Figure 1 This is a diagram illustrating a biopolymer synthesis system according to an embodiment of the present invention.

[0016] Figure 2 It is shown Figure 1 A diagram of the reactor.

[0017] Figure 3 This is a graph showing the pressure drop based on the flow rate, measured to compare the reactor of the present invention with a comparative example.

[0018] Figure 4 This is a graph showing the pressure drop determined by the reactor size according to the present invention.

[0019] Figure 5 and Figure 6 The graph shows the pressure drop measured by comparing the reactor of the present invention with that of a comparative example.

[0020] Figure 7 Showing the appearance of the core substrate before coating, Figure 8 The appearance of the composite solid support is shown.

[0021] Figures 9 to 11 The high-performance liquid chromatography (HPLC) chromatogram of ACP synthesized using a specific example of a composite solid support and according to the flow rate is shown. Detailed Implementation

[0022] An embodiment of the present invention provides a biopolymer synthesis system, the biopolymer synthesis system comprising: a mixing storage tank for mixing at least one of amino acids, reagents, and additives; a reactor into which fluid discharged from the mixing storage tank flows; a temperature regulator disposed between the mixing storage tank and the reactor and regulating the temperature of the fluid flowing into the reactor; and a pump for providing driving force to cause the fluid to flow from the mixing storage tank to the reactor, the reactor comprising: a column having an inlet for the fluid inflow and an outlet for the fluid outflow; and a composite solid support disposed inside the column.

[0023] Additionally, the composite solid support may include: a core substrate; and a functional coating located on the core substrate.

[0024] In addition, the composite solid support is self-standing when the fluid flows through the column.

[0025] In addition, in the composite solid support, the loading density of the functional coating can be from 0.01 mmol / g to 2 mmol / g.

[0026] Furthermore, when the fluid flows through the reactor, at a flow rate of 40 C.V / min or less, the pressure drop between the inlet and the outlet can be less than 10 bar. Specifically, the pressure drop can be measured based on the pressure difference generated at the inlet and outlet of an empty column. Additionally, the flow rate can be measured in terms of column volume per minute (column volume). Volume (CV) is used to represent the flow rate of the reactor, which is below 40 C.V / min, specifically below 40 C.V / min, below 30 C.V / min, below 20 C.V / min, 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 Under the following conditions, the pressure drop can be below 10 bar: 0.1 CV / min to 30 CV / min, 5 CV / min to 30 CV / min, 10 CV / min to 30 CV / min, 15 CV / min to 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.Meanwhile, within the stated flow rate range, the pressure drop can be below 10 bar, specifically, it can be below 10 bar, below 5 bar, below 3 bar, below 1 bar, below 0.5 bar, below 0.1 bar, below 0.05 bar, 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, 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.

[0027] Furthermore, the temperature of the fluid flowing into the reactor after passing through the temperature regulator can be below 120°C. The biopolymer system of the present invention can synthesize biopolymers with excellent purity and efficiency even within a variety of fluid temperature ranges. The fluid temperature can be below 120°C, specifically below 120°C, below 100°C, below 80°C, below 60°C, 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 to these ranges. Moreover, the temperature of the fluid used in biopolymer synthesis can be set without limitation.

[0028] Additionally, it may include a valve disposed between the reactor and the mixing storage tank, for discharging waste.

[0029] Additionally, it may include: a main pipeline for connecting the mixing storage tank, the pump, the temperature regulator, and the reactor; and a first branch pipeline branching from the main pipeline between the mixing storage tank and the reactor and connecting to the mixing storage tank.

[0030] In addition, the fluid can be circulated through the main pipeline.

[0031] Additionally, a solvent storage tank may be provided on the first branch pipeline.

[0032] In addition, the solvent storage tank can be connected to the valve and the mixing storage tank via the first branch pipeline.

[0033] Additionally, it may include: a sensor disposed on the main pipeline for sensing at least one of the temperature, pressure, and flow rate of the fluid flowing along the main pipeline.

[0034] Additionally, it may include a detector disposed on the main pipeline for detecting the synthesis process of the biopolymer.

[0035] Additionally, a second branch line may be included, branching off from the valve and used to discharge the waste. Specific Implementation Various modifications can be made to this invention, and the invention may have multiple embodiments. Specific embodiments will be exemplarily shown in the accompanying drawings and described in detail in the detailed description. The effects and features of the invention, as well as methods of implementing them, will become clear with reference to the detailed embodiments and drawings described below. However, the invention is not limited to the embodiments disclosed below, but can be implemented in many other forms.

[0037] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings. In the description with reference to the drawings, the same or corresponding constituent elements will be given the same reference numerals, and repeated descriptions thereof will be omitted.

[0038] In the following embodiments, the terms "first" and "second" are not used in a restrictive sense, but rather to distinguish one constituent element from another.

[0039] Unless the context clearly indicates otherwise, in the following embodiments, singular expressions include plural expressions.

[0040] In the following embodiments, terms such as "comprising" or "having" mean that the features or constituent elements described in the specification are present, but do not preclude the possibility of adding more than one other feature or constituent element.

[0041] In the following embodiments, when a membrane, region, constituent element, or other part is located "above" or "on top of" another part, this includes not only the case where it is directly above another part, but also the case where another membrane, region, constituent element, or other part exists in between.

[0042] In the accompanying drawings, the dimensions of the constituent elements may be exaggerated or reduced for ease of description. For example, the dimensions and thicknesses of each component shown in the figures are arbitrarily illustrated for ease of description, and therefore the invention is not necessarily limited to the figures shown.

[0043] In the following embodiments, when described as having membranes, regions, constituent elements, etc. connected, this includes not only the case where the membranes, regions, and constituent elements are directly connected, but also the case where another membrane, region, or constituent element exists in between, thus indirectly connecting them. For example, in this specification, when membranes, regions, constituent elements, etc. are electrically connected, this includes not only the case where the membranes, regions, and constituent elements are directly electrically connected, but also the case where another membrane, region, or constituent element exists in between, thus indirectly electrically connecting them. Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0044] In this specification, the term "loading density" refers to "the number of moles of functional groups provided per unit mass of composite solid support," and may refer to the biopolymer synthesis reaction sites according to the composition ratio of the polymer for biopolymer synthesis in an embodiment.

[0045] In one embodiment, the composite solid support may include: a structure core having a length of 1 mm or more and having a shape of one or more dimensions; and a functional coating surrounding the structure core, in which biopolymers can be synthesized.

[0046] In one implementation, the functional coating may have the property of swelling in a solvent.

[0047] In one embodiment, the surface and interior of the functional coating may contain functional groups. Specifically, the functional coating contains functional groups whose swelling properties in solvents facilitate the penetration of reactants into the interior of the functional coating due to concentration gradients. Thus, the functional groups on the surface and interior of the functional coating are easily exposed to the reactants, thereby exhibiting high synthesis efficiency even at low loading densities.

[0048] In one implementation, the functional group may include one or more selected from amino, carboxyl, hydroxyl, carbonyl, amino, thiol and phosphate groups.

[0049] In one implementation, the loading density of the reactive sites of the functional coating can be from 0.01 mmol / g to 2 mmol / g. Specifically, the loading density of the reaction sites in the functional coating can be 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 m 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 The values ​​are 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 0.7 mmol / g to 1 mmol / g, but are not limited to these.

[0050] In one embodiment, the functional coating's swelling rate per unit mass in water can be from 2 mL / g to 8 mL / g. Specifically, the functional coating's swelling rate per unit mass in water can be 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, but is not limited thereto. Simultaneously, the functional coating can also swell in solvents other than water, and depending on the type of solvent, it can have different swelling rates per unit mass.

[0051] In one embodiment, the core substrate may possess solvent resistance, thermal resistance, or both. Unlike the functional coating, the core substrate does not exhibit swelling or solvent solubility, thus enabling it to remain self-standing and function as a support for the composite solid structure even under high flow rate conditions. Furthermore, the core substrate is thermally resistant, allowing it to function as a support without alteration even during the synthesis of biopolymers at high temperatures.

[0052] In one embodiment, the core substrate may comprise one or more components selected from polymers, metals, and ceramics. Specifically, the polymer may comprise one or more components 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), polyvinylidene fluoride (PVDF), polyoxymethylene (PC), polycarbonate (PC), polyimide (PI), polyetheretherketone (PEEK), polyethersulfone (PES), polyphenylene ether (PPO), and polyphenylene sulfide (PPS), but is not limited thereto. The metal may comprise one or more components selected from stainless steel, titanium, nickel, tantalum, zirconium, and their alloys, but is not limited thereto. The ceramic may include, but is not limited to, one or more of the following: fused silica, alumina, zirconia, silicon carbide, silicon nitride, boron nitride, and titanium diboride.

[0053] In one implementation, the core substrate may be in one or more shapes selected from one-dimensional, two-dimensional and three-dimensional shapes.

[0054] In one implementation, the one-dimensional shape may include one or more selected from staple fiber, fiber, and 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 composed of lengths capable of generating biopolymer synthesis, and the composite solid support may be loaded in a wound shape within the reactor for use in the biopolymer synthesis reaction.

[0055] In one implementation, 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, polymer film, and mesh. As a specific example, the two-dimensional shape of the core substrate may include a shape in which multiple core substrates of the one-dimensional shapes are randomly arranged. Furthermore, through this random arrangement, pores are formed between the cores of the multiple one-dimensional shapes, and the two-dimensional shape can exhibit random porosity.

[0056] In one implementation, the three-dimensional shape may include open cell foam, a macropored sphere, or both.

[0057] In one implementation, the length of a cross-section of the core substrate can be 1 mm or more, and the functional coating can be large enough to participate in the synthesis of the desired biopolymer. As a specific example, when the core substrate is one-dimensional, the composite solid support can function as a support body by winding the core substrate into shapes such as fibers. Furthermore, when the core substrate is two-dimensional or three-dimensional, it can itself serve a supporting function, such as non-woven fabrics (two-dimensional) and foams (three-dimensional). Therefore, the length of a cross-section of the core substrate can be 1 mm or more, and theoretically, a one-dimensional core substrate of infinite length can also serve as a support body.

[0058] In one embodiment, the functional coating may comprise a polymer of one or more major 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 comprising a first monomer, a second monomer, and the active monomer. Alternatively, the functional coating of the present invention may be a polymer formed by polymerization with an active monomer that provides reaction sites primarily composed of one or more major monomers.

[0059] In one implementation, the first monomer may include one or more crosslinking agents selected from bisacrylamide crosslinking agents, methacrylamide, alkenyl-substituted triazine, and acrylate crosslinking agents. Specifically, the first monomer may include polyethylene glycol, such as polyethylene glycol diacrylate. Specifically, any substance in which the first monomer can react or bind with or to an active monomer that exposes functional groups within and / or on the surface of the functional coating can be used without limitation. As a non-limiting example, the first monomer may include a selection 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 (DALL). Phthalate (DAP) may include one or more of the following: 2-hydroxyethyl acrylate, 2-hydroxyethyl methacrylate (HEMA), acrylic acid (AA), methyl methacrylate (MMA), ethyl acrylate (EA), butyl acrylate (BA), glycidyl methacrylate (GMA), and vinyl acetate (VAc).

[0060] In one implementation, the active monomer is used to provide functional groups to the interior and / or surface of the functional coating, and may include one or more substances selected from acrylates, acrylamides and methacrylamides. As a non-limiting example, the active monomer may include a selection from N-(2-aminopropyl)methacrylamide hydrochloride, aminoethylmethacrylate hydrochloride (AEMA.HCl), 2-aminoethylmethacrylate (2-AEMA), N-(3-aminopropyl)methacrylamide hydrochloride (APMA), 4-aminostyrene, N-(4-aminophenyl)methacrylamide, N,N-dimethylaminoethylmethacrylate (DMAEMA), and N,N-dimethylaminopropylacrylamide. One or more of Acrylamide (DMAPAA) and N-(2-Aminoethyl)acrylamide hydrochloride.

[0061] 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 from a polymer of one or more of the main monomers and the active monomers. Alternatively, in the heterogeneous composite solid support, the functional coating, including a pulverized existing solid support, may be a shape coated on the core substrate. The existing solid support may include, but is not limited to, one or more selected from polystyrene / divinyl ether copolymer (PS / DVB), crosslinked polyethylene glycol polymer, poly-ε-lysine / sebacic acid copolymer, controlled-pore glass, amino-polyacrylamide-resin fiber, cellulose, and hydroxylated polypropylene.

[0062] In one implementation, the diameter (section) of the one-dimensional core substrate can be from 10 μm to 100 μm. Specifically, it can 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. When the diameter of the one-dimensional core substrate is less than 10 μm, the core substrate will agglomerate due to its excessive fineness, resulting in a reduction in mechanical properties. When the diameter exceeds 100 μm, the load density of the functional coating may be too low.

[0063] In one implementation example, the thickness (layer thickness) of the two-dimensional core substrate can be from 10 μm to 10 mm. Specifically, the thickness of the two-dimensional core substrate can be 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, 20 ... The ranges from 10 μm to 1 mm, but are not limited to these. These ranges include 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 0.5 mm, 350 μm to 0.5 mm, or 400 μm to 0.5 mm. When the thickness of the two-dimensional core substrate is less than 10 μm, it is difficult to maintain mechanical properties. When the thickness exceeds 10 mm, the polymerization reaction may not occur uniformly inside the functional coating.

[0064] In one embodiment, the porosity of the core substrate can be from 50% to 95%. When the core substrate is one-dimensional, pores can be formed by winding core substrates of fiber or other shapes. When the core substrate is two-dimensional or three-dimensional, pores may be present inside materials such as nonwoven fabrics or foams. With such pores, the porosity of the core substrate can 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 to these. There is no particular limitation on the pore size of the porous membrane substrate. When the porosity is less than 50%, the pores may be blocked even after the functional coating swells, preventing the synthesis of biopolymers within the functional coating. Furthermore, when the porosity exceeds 95%, it may be difficult to maintain mechanical properties.

[0065] In one implementation, the thickness of the functional coating can be from 0.1 μm to 1000 μm. Depending on the type of biopolymer required and whether the core substrate is one-dimensional, two-dimensional, or three-dimensional, the thickness of the functional coating can be selected. Specifically, the thickness of the functional coating can be 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 ... The thickness of the functional coating can range from 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, but is not limited to these. If the thickness of the functional coating is too thin, for example less than 0.1 μm, a problem arises where the loading density of the functional coating is too low. In addition, if the thickness of the functional coating is too thick, for example, exceeding 1000 μm, the following problem may occur: the functional coating itself can be prepared by an uneven polymerization reaction, resulting in a decrease in purity during the synthesis of biopolymers.

[0066] The composite solid support may not be a core-shell structure or a bead structure. Additionally, the solid support may not include a core in a core-shell structure.

[0067] In one implementation, the surface of the core substrate may be a hydrophilically treated surface. Specifically, the hydrophilic treatment can be performed by methods selected from, but not limited to, chemical methods (using surfactants or acidic solutions) and physical methods (including plasma treatment or ultraviolet irradiation). The functional coating may be formed on the hydrophilically treated core substrate.

[0068] In one implementation, the functional coating may further include a linker.

[0069] In one implementation, the linker may include one or more selected from Rink amide linkers, Wang linkers, 2-chlorotriphenylmethyl (2-Chlorotrityl, CTC) linkers, Sieber linkers, BAL linkers, 4-sulfamylbutyryl linkers, and hexamethylenebisacetamide (HMBA) (resistant to trifluoroacetic acid (TFA)) linkers.

[0070] In one implementation, functional groups may be exposed on the interior and / or surface of the functional coating, and these functional groups may be bonded to the linkers. Biopolymers comprising multiple monomer units can be synthesized by adding additional monomer units (amino acid residues) to the monomer units (amino acid residues) linked by the functional coating (functional group)-linker.

[0071] In one implementation, the biopolymer may include, but is not limited to, one or more selected from peptides, oligonucleotides and peptide nucleic acids (PNAs).

[0072] Figure 1 This is a diagram illustrating a biopolymer synthesis system according to an embodiment of the present invention.

[0073] Reference Figure 1 According to an embodiment of the present invention, a biopolymer synthesis system 1 can synthesize biopolymers by forming a circulating flow in a fluid containing at least one of amino acids, reagents and additives.

[0074] Below, “amino acid mixture” refers not only to the amino acids (multiples) used in this specification, but also to a liquid containing reagents (multiples), wherein the “amino acids” may be modified or unmodified, optionally pre-activated, and the amino acid mixture may also contain peptides.

[0075] Below, "SPPS" is an abbreviation for Solid Phase Peptide Synthesis, which is used to mean the production of peptides by adding amino acid residues to peptides or amino acids immobilized on a solid support (resin).

[0076] Below, "reagent" is used in the sense of coupling reagent, deprotection reagent, additive, base, and other reagents used in synthesis.

[0077] Below, "fluid" refers to the substance discharged from the storage tank and moving along the pipes of the biopolymer synthesis system, and depending on the process, it can be a variety of single substances or a mixture of substances.

[0078] As an example, the biopolymer synthesis system 1 is applicable to the flow-through process of solid phase peptide synthesis (SPPS) to sequentially synthesize at least one amino acid.

[0079] As one embodiment, the biopolymer synthesis system 1 may include: a mixing storage tank 100, a pump 200, a temperature regulator 300, and a reactor 400. Additionally, the biopolymer synthesis system 1 may also include: a valve 500, a solvent storage tank 500, a sensor SE, and a detector DE.

[0080] The main pipeline mL can provide a circulation line by connecting the mixing storage tank 100, pump 200, temperature regulator 300, and reactor 400. In addition, a valve 500 can be installed on the main pipeline mL, and a first branch pipeline SL1 and a second branch pipeline SL2 can be connected to the valve 500.

[0081] The mixing storage tank 100 provides space for the residence of fluids containing at least one of amino acids, reagents, and additives. An on / off valve (not shown) is provided at the outlet of the mixing storage tank 100 for regulating the type and flow rate of the fluid flowing into the main pipeline (mL).

[0082] The mixing storage tank 100 can mix amino acids, reagents, and additives flowing in from one or more reagent storage tanks (not shown) and solvent storage tanks. Operators can precisely adjust the type and flow rate of the fluid discharged from the mixing storage tank 100 according to each synthesis step by adjusting the opening volume of each chamber. As an example, in the following SPPS method of biopolymer synthesis steps, this fluid can flow into the mixing storage tank, be mixed and retained, and then flow into the reactor.

[0083] 1) Deprotection step: Deprotection reagent (piperidine).

[0084] 2) Washing steps: Solvent (DMF).

[0085] 3) Coupling steps: amino acid, coupling reagent (N,N-diisopropylcarbodiimide (DIC)), additive (ethyl 2-oxime cyanoacetate (Oxyma pure)).

[0086] 4) Washing steps: Solvent (DMF).

[0087] Pump 200 can be used to provide driving force to move the fluid along the main pipeline mL. Pump 200 can provide driving force to move the fluid from the mixing storage tank 100 to the reactor 400. Figure 1In this process, the pump 200 is located between the mixing storage tank 100 and the reactor 400 during the flow of fluid. However, since the fluid forms a circulating flow, the pump is not necessarily located between the mixing storage tank 100 and the reactor 400.

[0088] Pump 200 can be any type of device that provides suction and discharge force to a fluid. For example, pump 200 can be a gear pump, screw pump, vane pump, cap pump, piston pump, piston pump, diaphragm pump, centrifugal pump, etc.

[0089] In addition, Pump 200 can use all types of pumps, such as mechanical displacement micro pumps and electromagnetic motion micro pumps. Mechanical displacement micro pumps are pumps that use the movement of solids or fluids (such as gears or diaphragms) to induce fluid flow and generate a pressure difference; these can be diaphragm displacement pumps, fluid displacement pumps, and rotary pumps. Electromagnetic motion micro pumps are pumps that directly use electrical or magnetic energy for fluid movement; these can be electrohydrodynamic pumps (EHD), electroosmotic pumps, magnetohydrodynamic pumps, and electrowetting pumps.

[0090] Temperature regulator 300 sets the temperature of the fluid flowing into reactor 400 to a target temperature. Temperature regulator 300 is located downstream of mixing storage tank 100 and upstream of reactor 400, and is used to cool or heat fluids comprising one or more selected from amino acids, reagents, and additives. Figure 1 In the process of fluid flow, the temperature regulator 300 is located between the mixing storage tank 100 and the reactor 400. However, since the fluid forms a circulating flow, the temperature regulator is not necessarily located between the mixing storage tank 100 and the reactor 400.

[0091] The temperature regulator 300 can be a variety of devices that transfer heat to or receive heat from a fluid through heat exchange. For example, the temperature regulator 300 can be an electric heating device, an induction heater, or a microwave cavity.

[0092] As an example, the temperature regulator 300 can maintain the temperature of the fluid flowing into the reactor 400 at ambient temperature. The temperature regulator 300 can adjust the fluid temperature to below 120°C, specifically from 25°C to 100°C. Additionally, the temperature regulator 300 can adjust the fluid temperature to from 50°C to 80°C.

[0093] Figure 2 It is shown Figure 1 A diagram of the reactor.

[0094] Reference Figure 1 and Figure 2 The reactor 400 allows the fluid discharged from the mixing storage tank 100 to circulate.

[0095] The reactor 400 may be equipped with a column 410 and a composite solid support 420.

[0096] 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 be provided with a composite solid support 420.

[0097] Considering the type of biopolymer synthesized, the amount of biopolymer synthesized, the flow rate of the fluid, the velocity of the fluid, and the type of composite solid support, the column 410 can have various sizes.

[0098] The inlet 411 and outlet 412 of the column 410 can be located on the main pipeline mL, so that the fluid entering from the inlet 411 passes through the internal space of the column 410 and is discharged through the outlet 412. At this time, the fluid can pass through the composite solid support 420 located in the internal space of the column 410.

[0099] The composite solid support 420 can be disposed inside the column 410. The solid support 420 can be filled inside the column 410.

[0100] The composite solid support 420 can be installed in various positions and shapes within the internal space of the column 410. The composite solid support 420 can fill a predetermined position within the internal space of the column 410.

[0101] For example, the composite solid support 420 can be installed in the internal space of the column 410.

[0102] As an example, when the core substrate of the composite solid support 420 is in the form of wound filaments FM, fluid can pass through the spaces between the filaments FM. The flow path of the fluid can vary depending on the loading method of the composite solid support 420. As an example, when the composite solid support is loaded in a spiral coiled shape, the fluid can flow in the porous spaces created by the winding of the layers of the spiral composite solid support.

[0103] The composite solid support 420 may have a core substrate BM and a functional coating CO.

[0104] As an example, the core substrate BM can be formed by winding multiple filaments to create random porosity. The functional coating CO can be a polymer synthesized from biopolymers coated onto the core substrate to expose functional groups such as amine and carboxyl groups.

[0105] The core substrate BM can be 1 mm or longer and can be in one-dimensional or higher shapes. The core substrate BM can be selected from one or more shapes, including one-dimensional, two-dimensional, and three-dimensional shapes.

[0106] The core substrate BM may possess properties such as solvent resistance, thermal resistance, or a combination of both. Unlike the functional coating CO, the core substrate BM does not exhibit swelling or solvent solubility, thus enabling it to be self-standing and function as a support for the composite solid structure. Simultaneously, the core substrate FM possesses thermal resistance, allowing it to function as a support without alteration even during the synthesis of biopolymers at high temperatures.

[0107] Functional coatings can surround the core substrate (FM) and provide a biopolymer synthesis zone.

[0108] As an example, the functional coating CO may have the property of swelling in a solvent.

[0109] The surface and interior of the CO functional coating may contain functional groups. Specifically, the functional coating contains functional groups that, due to their swelling properties in solvents, readily allow reactants to penetrate into the interior of the functional coating due to concentration gradients. Thus, the functional groups on the surface and inside the functional coating are easily exposed to the reactants, thereby exhibiting high synthesis efficiency even at low loading densities.

[0110] Valve 500 is positioned between reactor 400 and storage tank 100 to set the waste discharge path. Valve 500 can adjust the flow direction of the fluid so that the waste generated after the reaction in reactor 400 is discharged through the second branch line SL2. As an example, valve 500 can be a four-way valve, which can be connected to mixing storage tank 100, reactor 400, solvent storage tank 600, and second branch line SL2.

[0111] Valve 500 regulates fluid circulation and waste discharge. During each step of SPPS synthesis, valve 500 can improve the synthesis rate of biopolymer by setting the circulation flow of the reactant stream, and discharge the remaining materials from each step through the second branch line SL2, thereby enabling the next step.

[0112] The solvent storage tank 600 can store solvents used for biopolymer synthesis and cleaning. A variety of solvents can be selected depending on the synthesis process.

[0113] Solvent storage tank 600 may be installed on the first branch line SL1. Solvent storage tank 600 may be connected to the first branch line SL1 so that the solvent moves to valve 500 or to mixing storage tank 100 according to the steps of SPPS synthesis.

[0114] Before the washing process in the synthesis process, any residue remaining in the pipeline from the previous step should be removed. At this time, the solvent discharged from the solvent storage tank 600 can be discharged through the first branch line SL1 for use in the main washing line mL.

[0115] The sensor SE can be installed on the main pipeline mL to sense at least one of the temperature, pressure, and flow rate of the fluid flowing along the main pipeline mL.

[0116] The detector DE can be installed on the main pipeline mL to detect the synthesis process of the biopolymer. As an example, the detector can be installed at at least one of the inlet and outlet of reactor 400 to detect the circulating fluid. Figure 1 In this process, the detector DE is located between the reactor 400 and the mixing storage tank 100 during the fluid flow. However, since the fluid forms a circulating flow, the detector is not necessarily located between the reactor 400 and the mixing storage tank 100.

[0117] The main pipeline mL can connect the mixing storage tank 100, pump 200, temperature controller 300 and reactor 400, and provides a fluid circulation path.

[0118] The first branch line SL1 can branch off from the main line mL between the mixing storage tank 100 and the reactor 400 and connect to the mixing storage tank 100. The first branch line SL1 connects the valve 500 and the mixing storage tank 100, and supplies solvent from the solvent storage tank 600 to the main line mL to remove residual substances on the main line mL.

[0119] The second branch line SL2 can branch off from valve 500 and is used to discharge waste.

[0120] Preferred embodiments are presented below to facilitate understanding of the present invention. However, these embodiments are provided only for the purpose of facilitating a better understanding of the present invention, and the scope of the present invention is not limited to these embodiments.

[0121] Example 1. Pressure drop analysis of the reactor based on flow rate Figure 3 This is a graph showing the pressure drop based on the flow rate, measured to compare the reactor of the present invention with a comparative example.

[0122] (Experimental equipment) - Column (Intertec empty cartridge column): 27 mL, ID / Height: 12.8 60mm.

[0123] - Pump: LEAD FLUID CT3001, Pump head: Fluid-o-Tech / MG209.

[0124] - Reagents used: Dimethylformamide (DMF, SAMJHUN), Rink-Amide-MBHA-resin (GL Biochme, 0.5 mmol / g).

[0125] - Pressure measuring equipment: SMC PSE560-C01 pressure sensor, SMC PSE200A digital pressure sensor controller.

[0126] As an example, 5.0 g (0.5 mmol, loading density: 0.1 mmol / g) of the composite solid support was placed in a column. As a comparative example, 1.0 g (0.5 mmol, loading density: 0.5 mmol / g) of particulate solid support (PS / DVB) was placed in a column, and then 20 mL of DMF solvent was added, and the composite solid support was allowed to swell for 30 minutes.

[0127] Then, connect the pump's discharge pipe to the column, and install pressure sensors at the inlet and outlet of the column, and connect each pressure sensor to the controller.

[0128] DMF was introduced into an empty column, a composite solid support column (Example), and a granular solid support column (Comparative Example) at a flow rate of 40 mL / min (1.48 CV / min), and the pressure at the inlet and outlet was measured. Then, the flow rate was changed 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), and the pressure at the inlet and outlet of the column was measured again.

[0129] The pressure drop values ​​generated at the inlet and outlet of the empty column are set as the baseline values ​​(Zero). Calculate: the additional pressure drop values ​​generated when the composite solid support is placed at the inlet and outlet of the column compared to the baseline values, and the additional pressure drop values ​​generated when the granular solid support is placed at the inlet and outlet of the column compared to the baseline values.

[0130] Table 1 Reference Figure 3 As shown in Table 1, even with increased flow rate, the pressure drop of the composite solid support of the present invention is very small. This is because the fluid can pass through the empty spaces of the composite solid support, thus the pressure drop is small even with increased flow rate, and the fluid's fluidity is increased. Conversely, when the flow rate increases, the existing particulate solid support (beaded solid-phase synthesis resin polymer (PS / DVB)) produces a large pressure drop, and the fluid does not drain when the flow rate reaches 300 mL / min. This is because the existing particulate solid support aggregates and fills the space within the column when the flow rate increases, reducing the space through which the fluid can pass, thus significantly reducing the pressure measured at the outlet and decreasing the fluid's fluidity.

[0131] Example 2. Pressure drop analysis of composite solid supports based on column dimensions Figure 4 This is a graph showing the pressure drop determined by the reactor size according to the present invention.

[0132] (Experimental equipment) -A column (Intertec empty cartridge column): 27mL, ID / Height: 12.8 60mm.

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

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

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

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

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

[0138] Load 5 g of the composite solid support into column A and add 20 mL of DMF solvent, allowing it to swell for 30 minutes. Load 20 g of the composite solid support into column B and add 80 mL of DMF solvent, allowing it to swell for 30 minutes. Load 70 g of the composite solid support into column C and add 300 mL of DMF solvent, allowing it to swell for 30 minutes.

[0139] Connect the pump's discharge pipe to each column, install pressure sensors at the inlet and outlet of each column, and connect each pressure sensor to the controller.

[0140] DMF was introduced into columns A, B, and C at a flow rate of 0.5 column volume (mL) / min, and the pressure at the inlet and outlet was measured. Then, the flow rate was 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 pressure at the inlet and outlet of the columns was measured again.

[0141] The pressure drop values ​​generated at the inlet and outlet of each empty column are set as the baseline value (Zero). The additional pressure drop values ​​generated when the composite solid support is placed at the inlet and outlet of columns A, B, and C are calculated compared to the baseline value.

[0142] Table 2 Reference Figure 4 As shown in Table 2, even with changes in the volume of the column, the composite solid support of the present invention exhibits minimal pressure drop of the fluid under various flow rate conditions. This is because the fluid can pass through the empty spaces of the composite solid support, thus the pressure drop remains small even with increased flow rate, and the fluid's mobility increases. Therefore, the composite solid support of the present invention maintains a consistently low pressure drop regardless of the volume of the column.

[0143] Example 3. Pressure drop analysis of composite solid supports based on temperature Figure 5 and Figure 6 The graph shows the pressure drop measured by comparing the reactor of the present invention with that of a comparative example.

[0144] At the same flow rate, the faster the pump rotates, the greater the back pressure. This method allows for comparison of the back pressure of different solid supports by measuring the pump's rotational speed at a specific flow rate.

[0145] The following comparisons were made: a 27 mL column loaded with the composite solid support (0.5 mmol) used in this 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).

[0146] As shown in Table 3 and Figure 5 As shown, at 25°C, using DMF solvent, the RPM values ​​of the pump were recorded at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 mL / min.

[0147] When using DMF at 25°C, there was no significant difference in pump speed between the column loaded with composite solid support (Example) and the empty column (Comparative Example 1). In contrast, the column loaded with particulate solid support (Comparative Example 2) stopped pumping due to high back pressure at 250 mL / min.

[0148] Table 3 As shown in Table 4 and Figure 6 As shown, at 50°C, using DMF solvent, the pump RPM values ​​were recorded at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 mL / min. When using DMF at 50°C, the decrease in solution viscosity led to a decrease in back pressure. However, with the particulate solid support (Comparative Example 2), the pump stopped operating due to high back pressure at a flow rate of 350 mL / min.

[0149] Table 4 As shown in Table 5, the pump RPM values ​​were recorded at flow rates of 50, 100, 150, 200, 250, 300, 350, and 400 mL / min using DMF solvent at 70°C. When using DMF at 70°C, the decrease in solution viscosity led to a decrease in back pressure. However, with the particulate solid support (Comparative Example 2), the pump stopped operating due to high back pressure at a flow rate of 350 mL / min.

[0150] Table 5 Furthermore, the pump rotation frequency was measured at room temperature using DMF (0.1 M oxymapure) of piperidine, commonly used in 20% solid-phase peptide synthesis reactions. Even at this rate, the pump stopped operating at 250 mL / min due to high back pressure. This confirms that the composite solid support of the present invention has lower back pressure than existing granular solid supports. Considering that the volume and weight of the composite solid support increase with the length of the synthesized peptide, leading to increased back pressure, unlike existing granular solid supports, the composite solid support of the present invention can improve the synthesis performance of a flow reactor by increasing fluid flowability.

[0151] An embodiment of the present invention provides a flow reactor and a biopolymer synthesis system including the same, which can improve the efficiency of the synthesis reaction.

[0152] In existing particulate solid-phase synthesis, resin polymers aggregate at high flow rates, completely filling the internal space of the column and reducing reactor fluidity. This results in a high pressure drop at the reactor outlet, leading to high back pressure. High back pressure prevents a significant increase in the synthesis scale of the system.

[0153] An embodiment of the present invention provides a flow reactor and a biopolymer synthesis system including the same, which improves the reactor's fluidity through a composite solid support. Therefore, the reactor generates a low pressure drop, thereby achieving a low back pressure and enabling increased synthesis scale.

[0154] Specifically, under the same pressure conditions, when the back pressure is low, the flow rate decreases (losses) and thus promotes mixing and diffusion between the reactants and the reaction solution within the column. Biopolymer synthesis systems exhibit improved flow dynamics with increasing flow rate, thereby enhancing reaction efficiency and yield.

[0155] Furthermore, reduced back pressure decreases wear and mechanical stress on internal reactor components, thereby improving the overall system durability. Additionally, pumps capable of providing various pressures can be used, increasing availability and choice. In particular, high reaction efficiency and yield are achieved even when using low-pressure pumps.

[0156] In addition, if the back pressure is reduced, the damage to the pipes and connections through which the fluid flows will be reduced, which can improve the durability, process stability and reliability of the system by reducing fluid leakage.

[0157] Furthermore, if the back pressure is reduced, the fluid flowing through the reactor can be precisely controlled, thus improving the reproducibility of the reaction process and synthesizing biopolymers of a certain quality.

[0158] Example 4. Preparation of composite solid support (1) Preparation of polymers for functional coatings Experiments were conducted using a previously confirmed flow reactor to determine the synthetic properties of the biopolymer. First, the polymer for the functional coating used in the composite solid support was prepared. The reagents shown in Table 6 below were used, and each reagent was mixed and stirred in a solvent until homogeneously dissolved. At this point, the composition of the functional coating polymer only needed to meet the specified mass ratios (parts by weight), and the solvent could be used arbitrarily, as drying would follow. In the experiments, the total amount of each reagent and solvent (DW) reached 100 g [in the total (reagent + solvent), the solvent could be used in the range of 10 wt% to 90 wt%].

[0159] Table 6 Then, 0.5 g of 2-hydroxy-2-methylpropiophenone, acting as a curing initiator, was added to the solution and mixed uniformly to prepare a functional coating solution. The functional coating solution was then uniformly coated onto a petri dish and treated with 10 mJ / cm² water. 2 Up to 1000mJ / cm 2 The polymer was thoroughly irradiated with ultraviolet energy to cure it. After curing, the reaction residue was removed with ethanol to obtain functional coated polymer discs. The mass of the dried functional coated polymer was 45.7 g. The prepared functional coated polymer was rapidly frozen with liquid nitrogen and then pulverized using a mortar and pestle. Then, using 100 and 200 mesh sieves, functional coated polymer particles with a particle size range (74~149 μm) of 100~200 mesh were obtained.

[0160] (2) Preparation of composite solid supports for biopolymer synthesis To prepare the composite solid support, 20g of the functional coating solution (before curing) from Example 4 (1) and the core substrate (polypropylene, spunbond nonwoven fabric, 40g / m²) were prepared. 2 The core substrate is fully immersed in the functional coating solution, and after being removed from the solution, it is cured in a UV curing chamber at 10 mJ / cm². 2 ~1000mJ / cm 2 It is fully irradiated with ultraviolet energy.

[0161] After curing, ethanol was used to remove reaction residues from the coated core substrate, followed by drying. A composite solid support with a functional coating polymer coated on the 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 follows Figure 7 As shown, the appearance of the composite solid support is as follows Figure 8 As shown.

[0162] Example 5. Synthesis of biopolymers using composite solid supports Compared to batch automated synthesizers (CEM Liberty, USA), this study investigates whether peptides can be synthesized using a SPPS method in a biopolymer synthesis system comprising a column-shaped flow reactor containing a solid support for biopolymer synthesis, as a specific example. The study examines whether there are differences in the purity and yield of the synthesized peptides.

[0163] (1) Synthesis of peptides using an intermittent automated synthesizer After adding 0.1 mmol of existing solid-phase synthesis particulate solid support and a specific example of composite solid support to an automated synthesizer, the synthesis reaction of the acyl carrier protein (ACP) model peptides (65-74) of SEQ ID NO.1 was carried out in the same manner under the same conditions as in Table 7 below.

[0164] SEQ ID NO.1: VQAAIDYING.

[0165] Table 7 The particulate solid support and composite solid support after the peptide synthesis reaction were thoroughly washed with ethanol and dried. Then, they were subjected to a cleavage reaction for 2 hours using a cleavage solution (95% TFA, 2.5% TSI, 2.5% DW). After precipitation with cold diethyl ether, the solid was obtained by centrifugation, washed twice again with 15 mL of cold diethyl ether, and then thoroughly dried under reduced pressure. The purity and yield of the dried peptide were determined by high-performance liquid chromatography (HPLC). The results of the determination of the purity and yield of the dried peptide are shown in Table 8.

[0166] Table 8 In peptide synthesis analysis using a commercially available intermittent SPPS synthesizer (Liberty CEM, USA) with particulate solid supports and a specific example of a biopolymer composite solid support, the results showed that the yield and purity of ACP peptides were improved in a specific example of a composite solid support compared to particulate solid supports.

[0167] (2) Synthesis of peptides using a continuous flow column synthesizer To compare the synthetic performance of particulate solid supports and homogeneous composite solid supports in a flow reactor, 0.5 mmol of particulate solid support and composite solid support were added to 27 mL columns, respectively. The amino acid types and reaction steps were then varied under the conditions shown in Table 9 below to synthesize the acyl carrier protein (ACP) model peptides (65-74) of SEQ ID NO. 1. The dried mass was then measured and the yield calculated. The purity of the synthesized peptides was measured using high-performance liquid chromatography (HPLC).

[0168] Table 9 Using a composite solid support, the synthesis in a flow reactor achieved a purity of 95.8% and a yield of 93.5%, demonstrating superior performance in both purity and yield compared to batch reactors. Conversely, when synthesizing ACP (65-74) peptides in a flow reactor using particulate solid supports, column clogging (backpressure issue) occurred, preventing normal synthesis. This is because the particulate solid support swells and clogs the flow reactor column. The composite solid support, with a functional coating on the core substrate, does not exhibit this problem and is therefore more suitable for flow reactors. This experiment confirms that the composite solid support is suitable for both batch and flow synthesis systems compared to existing particulate solid supports. In particular, when used in flow synthesis systems, the yield and purity of ACP peptides exceed 90%, demonstrating significantly superior efficiency in biopolymer synthesis.

[0169] Example 6. Analysis and evaluation of changes in the yield and purity of synthesized peptides based on the flow rate of biopolymers in a flow reactor. As shown in Example 5, the yield and purity of the synthesized peptides based on the flow rate were confirmed using a composite solid support. ACP was synthesized using the same method as in Example 5, and the purity and yield of the synthesized ACP peptides are shown in Table 10 below. Figures 9 to 11 middle.

[0170] Table 10 Figures 9 to 11The HPLC chromatogram of ACP synthesized using a specific example of a composite solid support for biopolymer synthesis and according to the flow rate is shown. When using a specific example of a solid support for biopolymer synthesis, it can be confirmed that the purity and yield of the peptide increase with the increase of the flow rate of the column synthesizer.

[0171] The results indicate that the flow reactor containing the column is a more suitable reactor for a specific example of a composite solid support.

[0172] As described above, the present invention has been illustrated with reference to the embodiments shown in the accompanying drawings, but these are merely exemplary, and it will be understood by those skilled in the art that various modifications and equivalent embodiments can be derived therefrom. Therefore, the true scope of protection of the present invention is determined by the technical concept of the appended claims.

Claims

1. A biopolymer synthesis system, comprising: a mixing reservoir for mixing at least one of an amino acid, a reagent, and an additive; a reactor into which a fluid discharged from the mixing reservoir flows; a temperature regulator disposed between the mixing reservoir and the reactor and regulating a temperature of the fluid flowing into the reactor; and a pump for providing a driving force for the fluid to flow from the mixing reservoir to the reactor, wherein the reactor comprises: a column having an inlet into which the fluid flows and an outlet from which the fluid is discharged, and a composite solid support disposed inside the column.

2. The biopolymer synthesis system according to claim 1, wherein the composite solid support comprises: a core substrate; and a functional coating layer on the core substrate.

3. The biopolymer synthesis system according to claim 2, wherein the composite solid support is self-supporting when the fluid flows through the column.

4. The biopolymer synthesis system according to claim 2, wherein a loading density of the functional coating layer in the composite solid support is 0.01 mmol / g to 2 mmol / g.

5. The biopolymer synthesis system according to claim 1, wherein a pressure drop between the inlet and the outlet is 10 bar or less at a flow rate of 40 C.V / minute or less when the fluid flows through the reactor.

6. The biopolymer synthesis system according to claim 1, wherein a temperature of the fluid flowing into the reactor after passing through the temperature regulator is 120°C or less.

7. The biopolymer synthesis system according to claim 1, further comprising a valve disposed between the reactor and the mixing reservoir and discharging a waste.

8. The biopolymer synthesis system according to claim 1, further comprising: a main line connecting the mixing reservoir, the pump, the temperature regulator, and the reactor; and a first branch line branched from the main line between the mixing reservoir and the reactor and connected to the mixing reservoir.

9. The biopolymer synthesis system according to claim 8, wherein the fluid circulates through the main line.

10. The biopolymer synthesis system according to claim 8, further comprising: a solvent reservoir disposed on the first branch line.

11. The biopolymer synthesis system according to claim 10, wherein the solvent reservoir is connected to the valve and the mixing reservoir through the first branch line.

12. The biopolymer synthesis system according to claim 8, further comprising: a sensor disposed on the main line and sensing at least one of a temperature, a pressure, and a flow rate of the fluid flowing along the main line.

13. The biopolymer synthesis system according to claim 8, wherein ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ Also included is a detector disposed on the main line for detecting the progress of synthesis of the biopolymer.

14. The biopolymer synthesis system of claim 7, wherein, Also included is a second branch line branching from the valve and for discharging the waste.

Images