Composite solid support for biological polymer synthesis

Abstract

The present application relates to a composite solid support for synthesizing a biological polymer with high purity, high yield, and in a large quantity; a preparation method therefor; and a method for synthesizing a biological polymer by using same.

Description

Composite solid supports for the synthesis of biological polymers

[0001] The present invention relates to a composite solid support for synthesizing a biological polymer with high purity, high yield, and in large quantities, a method for producing the same, and a method for synthesizing a biological polymer using the same.

[0002] The process of synthesizing biological polymers, such as peptides or oligonucleotides, consists of repetitive unit processes. These unit processes consist of coupling reactions and deprotection reactions. The coupling reaction covalently bonds monomers, such as amino acids or nucleotides, one by one to synthesize a biological polymer of the target sequence. The deprotection reaction removes the protecting groups attached to the monomers to prevent side reactions during the coupling reaction and prepare for the next coupling reaction. Biological polymer synthesis reactions require the sequential addition of monomers. Therefore, after the coupling and deprotection reactions, it is important to remove unreacted and by-products and isolate only the desired product with high yield and purity.

[0003] Methods for synthesizing biological polymers include liquid-phase synthesis and solid-phase synthesis. Liquid-phase peptide synthesis (LPPS) uses precipitation, crystallization, or chromatography to separate the products of each conjugation and deprotection reaction. These separation methods are time-consuming and costly, and have limitations, with losses exceeding 1.0% in each unit process. This loss during the separation process leads to a sharp decline in synthetic yield as the biological polymer sequence lengthens.

[0004] Solid-phase peptide synthesis (SPPS) was introduced to overcome these limitations, centered on the research of Bruce Merrifield (Nobel Prize in Chemistry, 1984). The hallmark of solid-phase synthesis is the step-by-step synthesis of a biological polymer of a specific sequence on a solid support, with physical separation at each step to remove unreacted products and by-products, resulting in high product yields. A representative separation method is filtration, which utilizes a filter with a filtration diameter that allows the reaction solution to pass through but not the solid support. Typically, the unit process loss rate of the solid support when using a separation method using a filter is less than 0.1%.

[0005] Solid-state synthesis has become the standard for the synthesis of long-length biological polymers with high yields, and extensive research has been conducted to develop solid supports with diverse and improved properties. Solid-state support development can be broadly categorized into single-material and composite materials, and particulate or non-particulate forms, depending on their morphology. Single-material supports are composed of a single substance, exhibiting uniform physicochemical properties and offering the advantages of simple manufacturing and molding processes.

[0006] Representative single-material solid supports include particulate polystyrene / divinylbenzene copolymer (hereinafter PS / DVB), crosslinked polyethyleneglycol, poly-ε-lysine / sebacic acid, controlled pore glass (hereinafter CPG), non-particulate amino-polyacrylamide resin fiber, cellulose, and hydroxylated polypropylene (PP).

[0007] The most commonly used solid support in the field of solid-state synthesis of biological polymers commercially is PS / DVB, which has a network molecular structure in which linear polystyrene (PS) polymer chains are cross-linked with 1-2% by mass of divinylbenzene (DVB). PS / DVB is widely used due to its low cost, physicochemical stability, and wide loading density (or reaction site density), but it has the limitation of being a hydrophobic support. Due to the nature of the hydrophobic support, PS / DVB exhibits very low volume swelling characteristics in polar solvent conditions such as water or alcohol, and aggregation easily occurs during the synthesis of hydrophobic biological polymers.

[0008] A composite solid support is a new material that physically or chemically combines two or more different materials to complement each other's shortcomings and maximize their strengths. Representative composite solid supports include particulate-type PEG-grafted PS / DVB copolymer (PEG@PS / DVB) and magnetic nanoparticles (MNP), and non-particulate-type hydroxypropyl acrylate-coated polypropylene membranes (HPA-PP) and peptide synthetic films (PSF).

[0009] PEG@PS / DVB copolymer is a composite material manufactured by grafting hydrophilic PEG onto a PS / DVB particle-shaped polymer support with excellent physical strength and hydrophobic properties. It has the advantages of being able to effectively synthesize hydrophobic peptides that are difficult to synthesize using conventional PS / DVB solid supports, and providing high volume swelling characteristics and a wide range of loading densities in various solvent environments. However, the complexity of the grafting process leads to high manufacturing costs.

[0010] MNPs are particulate composite supports manufactured by coating magnetic materials, typically iron oxide, with silica (SiO2) or amino dextran. The magnetic material serves as the composite matrix, providing the magnetic function and particle shape, while the silica or amino dextran serves as the functional matrix, providing reaction sites. Due to the nature of the functional matrix, it has no or minimal volume swelling properties and relies on surface reactions, resulting in a very low loading density.

[0011] Peptide therapeutics generate billions of dollars in annual sales in areas such as diabetes, obesity, and oncology, and are expanding into the development of novel drugs for emerging diseases such as cardiovascular and neurodegenerative diseases. Currently, the short half-life of peptides has been addressed through methods such as the introduction of unnatural amino acids or fatty acid linkers with albumin-binding functionality. A variety of peptide therapeutics with dosing cycles exceeding one week are already on the market, and several oral delivery candidates are undergoing clinical trials with the US Food and Drug Administration (FDA). Demand for peptide therapeutics is growing by nearly 10% annually, necessitating new synthetic platforms to meet this increased productivity.

[0012] Additionally, various types of reactors are being studied to increase the production capacity for the synthesis of various biological polymers, including peptides. Representative reactors include batch-type reactors and flow-type reactors.

[0013] Traditional batch reactors, the oldest reactor type, offer the advantage of easy access to references and data. However, as reactor capacity increases for large-scale synthesis, the ratio of reactor surface area to unit volume decreases rapidly, leading to a structural limitation: increased temperature variation between the reactor surface and the interior. To minimize temperature variation, peptide synthesis reactions using large batch reactors (100 liters or more) are typically conducted at room temperature. This leads to very long reaction times and lower yields and purity. Furthermore, uneven mixing in the reactor can also reduce synthesis yield and purity.

[0014] Flow reactors are a relatively new type of reactor. Unlike batch reactors, they offer a very high surface area-to-volume ratio, resulting in very small temperature fluctuations and uniform mixing of the reaction solution. Furthermore, they offer several advantages, including a smaller volume than batch reactors, the ease of real-time reaction analysis through the introduction of in-line detectors, and the ease of process automation.

[0015] Representative methods for synthesizing and producing biological polymers using flow reactors include packed particulate solid support columns (PC) and volume-adjustable packed particulate solid support columns (VA-PC). These methods, which use flow reactors for solid-phase synthesis of biological polymers, are known to significantly shorten the required reaction time compared to batch reactors, while also providing higher yields and purities compared to batch reactors, as the reaction is usually performed at high temperatures of over 50°C. However, it has been reported that the backpressure increases to nearly 50 bar when using PC, and that the flow rate of the reactor must be used slowly to maintain a low backpressure when using VA-PC.

[0016] When the back pressure of a flow reactor is high or the flow rate is low, the temperature difference within the reactor increases and non-uniform mixing occurs, thereby negating the advantages of the flow reactor. In flow reactors using PC or VA-PC, the back pressure increases as the amount of solid support loaded increases or the flow rate increases. Therefore, these methods are not suitable for mass production of biological polymers using large amounts of solid support, but are suitable for small-scale production using small amounts of solid support.

[0017] Existing solid supports have limitations in many respects. First, solid supports lacking volumetric swelling properties in solvents only allow reactions to occur at a small number of exposed reaction sites, making them suitable for diagnostic purposes such as screening rather than production. CPG, MNP, HPA-PP, and PSF were each manufactured using small particle sizes or membrane or film coatings to increase surface area, thereby increasing loading density. However, these methods failed to achieve significant levels of synthetic efficiency.

[0018] Particulate solid supports have limited reaction rates due to their small surface area per unit volume. In solid supports with volumetric swelling properties, reactants in the liquid phase diffuse from the outside to the inside of the solid support due to concentration differences, and reactions can also occur at reaction sites within the solid support. Consequently, they possess a structural characteristic in which reactivity decreases with distance from the solid support surface. Therefore, particulate solid supports with relatively small surface area per unit volume face structural limitations in terms of reactant diffusion and reaction rates.

[0019] Furthermore, particulate solid supports have structural limitations that make them inappropriate for flow reactors, making them inherently self-supporting. Particulate solid supports, due to their inherent nature as particles, are subject to external forces, such as gravity or fluid flow. Consequently, the flow of the reaction solution within the flow reactor causes packing, which in turn increases friction and back pressure due to the concomitant decrease in inter-particle pores within the solid support. This increased friction and back pressure generate frictional heat within the packed solid support and lower flow rates, reducing the inherent advantages of flow reactors, such as temperature and mixing uniformity.

[0020] However, the problems that the present invention seeks to solve are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0021] The first aspect of the present invention provides a composite solid support for synthesizing biological polymers, comprising a core substrate having a length of 1 mm or more and a shape of one dimension or more; and a functional coating positioned on the core substrate, wherein a biological polymer is synthesized on the functional coating.

[0022] The second aspect of the present invention provides a use for synthesizing a biological polymer using a composite solid support, wherein the composite solid support comprises a core substrate having a length of 1 mm or more and a shape of one or more dimensions; and a functional coating positioned on the core substrate, wherein a biological polymer is synthesized from the functional coating.

[0023] The third aspect of the present invention provides a method for producing a composite solid support for synthesizing biological polymers, comprising the steps of: preparing a functional coating solution by mixing a main monomer, an active monomer, an initiator, and a solvent; impregnating a core substrate having a length of 1 mm or more and a shape of one dimension or more into the functional coating solution; and polymerizing the functional coating solution applied to the core substrate.

[0024] A fourth aspect of the present invention provides a method for synthesizing a biological polymer, comprising the steps of: coupling a first amino acid residue having a protecting group linked to a composite solid support according to the first aspect; removing the protecting group; and coupling a second amino acid residue having a protecting group linked to the N-terminus or C-terminus of the first amino acid, from which the protecting group is removed.

[0025] The composite solid support of the present invention is characterized by having a functional coating having volume swelling properties under various solvent conditions surrounding or attached to a core substrate, and providing a large number of reaction sites for mass synthesis of biological polymers such as peptides with high purity and high yield.

[0026] The composite solid support of the present invention comprises a self-supporting core substrate, and thus does not move with the flow of the reaction solution within a flow reactor. Furthermore, unlike a case where a particulate solid support is supported, which is not self-supporting, the composite solid support exhibits the characteristic of inducing a very small increase in back pressure within the reactor. Therefore, the composite solid support of the present invention can be applied to both batch and flow reactors for the synthesis of biological polymers.

[0027] Since the loading density and the purity and yield of biological polymer synthesis can be controlled through the selection of monomers in manufacturing a functional coating that provides a reaction site, the composite solid support of the present invention has the characteristic of being usable for the synthesis of various biological polymers such as peptides, oligonucleotides, and peptide nucleic acids (PNA).

[0028] Figure 1 is a graph comparing the expansion rate of a polymer for functional coating according to one embodiment with the expansion rate of a conventional particle-type solid-state synthetic polymer (PS / DVB).

[0029] Figure 2 is a photograph showing the appearance of a core substrate before coating (a) and the appearance of a composite solid support formed with a functional coating (b) used in the manufacture of a composite solid support according to one embodiment.

[0030] FIG. 3 is a confocal laser scanning microscope (CLSM) image (a) and an SEM image (b) showing the formation of a functional coating on a composite solid support manufactured using a core substrate on which a hydrophilic coating (pretreatment) has been performed according to one embodiment.

[0031] Figure 4 is a confocal laser scanning microscope (CLSM) image (a, b) and an SEM image (c, d) showing the formation of a functional coating on a composite solid support manufactured using a core substrate that was not subjected to a hydrophilic coating (pretreatment) according to one embodiment.

[0032] Figure 5a is an image showing a schematic diagram of the appearance and shape of a homogeneous composite solid support manufactured according to one embodiment.

[0033] Figure 5b is an image showing a schematic diagram of the appearance and shape of a heterogeneous composite solid support manufactured according to one embodiment.

[0034] Figure 5c is an image showing a schematic diagram of the appearance and shape of a particle-type solid support used in conventional solid-state synthesis.

[0035] Figure 6 is a graph comparing the coupling yield according to the coupling reaction time of a conventional particle-type solid support and a homogeneous / heterogeneous composite solid support according to an embodiment.

[0036] Figure 7a is a graph showing the purity and yield of a model peptide, Acyl Carrier Protein (ACP) (65-74), synthesized in a batch reactor using a homogeneous composite solid support according to one embodiment.

[0037] Figure 7b is a graph confirming the purity and yield of ACP (65-74) synthesized in a batch reactor using a conventional particle-type solid support.

[0038] Figure 8 is a graph confirming the purity and yield of ACP (65-74) synthesized in a flow reactor using a homogeneous composite solid support according to one embodiment.

[0039] Figure 9a is a graph showing the purity and yield of ACP (65-74) synthesized at a flow rate of 100 mL / min in a flow reactor using a composite solid support according to one embodiment.

[0040] Figure 9b is a graph showing the purity and yield of ACP (65-74) synthesized at a flow rate of 200 mL / min in a flow reactor using a composite solid support according to one embodiment.

[0041] Figure 9c is a graph showing the purity and yield of ACP (65-74) synthesized at a flow rate of 300 mL / min in a flow reactor using a composite solid support according to one embodiment.

[0042] Figure 10 is a graph showing the purity of ACP (65-74) synthesis according to the functional coating content (thickness and mass) of a composite solid support according to an embodiment.

[0043] Throughout this specification, when a part is said to be “connected” to another part, this includes not only cases where it is “directly connected” but also cases where it is “electrically connected” with another element in between.

[0044] Throughout this specification, when it is said that an element is “on” another element, this includes not only cases where the element is in contact with the other element, but also cases where another element exists between the two elements.

[0045] Throughout this specification, whenever a part is referred to as "comprising" a component, unless otherwise specifically stated, this does not exclude other components but rather implies the inclusion of additional components. The terms "about," "substantially," and the like, as used throughout this specification, are used to indicate the numerical values ​​or approximate values ​​of the components when manufacturing and material tolerances inherent to the meanings stated, and are used to prevent unscrupulous infringers from unfairly exploiting disclosures that contain precise or absolute values ​​to facilitate understanding of this specification.

[0046] The terms “step of” or “step of” as used throughout this specification do not mean “step for”.

[0047] Throughout this specification, the term “combination(s) thereof” included in the expressions in the Makushi format means one or more mixtures or combinations selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

[0048] Throughout this specification, references to “A and / or B” mean “A or B, or A and B.”

[0049] Throughout the present specification, the description of “loading density” means “the number of moles of functional groups provided per unit mass of the composite solid support,” and may refer to the density of the synthetic reaction sites of the biological polymer.

[0050] Throughout this specification, reference to “biological polymer” means “any molecule comprising at least two chemically linked monomer units, which is a sequence-defined biopolymer.”

[0051] Hereinafter, implementation examples and embodiments of the present invention will be described in detail with reference to the attached drawings. However, the present invention may not be limited to these implementation examples and embodiments and drawings.

[0052]

[0053] The first aspect of the present invention provides a composite solid support for synthesizing biological polymers, comprising a core substrate having a length of 1 mm or more and a shape of one dimension or more; and a functional coating positioned on the core substrate, wherein a biological polymer is synthesized on the functional coating.

[0054] The second aspect of the present invention provides a use for synthesizing a biological polymer using a composite solid support, wherein the composite solid support comprises a core substrate having a length of 1 mm or more and a shape of one or more dimensions; and a functional coating positioned on the core substrate, wherein a biological polymer is synthesized from the functional coating.

[0055] In one embodiment of the present invention, the functional coating may have a property of swelling in a solvent.

[0056] In one embodiment of the present invention, the functional coating may include functional groups on the surface, interior, or both. Specifically, the functional coating includes functional groups, and due to its swelling property in a solvent, the reactant is easily able to penetrate into the functional coating due to a concentration gradient. This allows the functional groups on the surface and interior of the functional coating to be easily exposed to the reactant, thereby achieving high synthesis efficiency.

[0057] In one embodiment of the present invention, the functional group may include at least one selected from an amine group, a carboxyl group, a hydroxyl group, a carbonyl group, an amino group, a thiol group, and a phosphoric acid group.

[0058] In one embodiment of the present invention, the loading density of the functional coating may be 0.01 mmol / g to 2 mmol / g. Specifically, the loading density of the reaction site of the functional coating is 0.01 mmol / g to 2 mmol / g, 0.05 mmol / g to 2 mmol / g, 0.1 mmol / g to 2 mmol / g, 0.3 mmol / g to 2 mmol / g, 0.5 mmol / g to 2 mmol / g, 0.7 mmol / g to 2 mmol / g, 1 mmol / g to 2 mmol / g, 0.01 mmol / g to 1.7 mmol / g, 0.05 mmol / g to 1.7 mmol / g, 0.1 mmol / g to 1.7 mmol / g, 0.3 mmol / g to 1.7 mmol / g, 0.5 mmol / g to 1.7 mmol / g, 0.7 mmol / g to 1.7 mmol / g, 1.0 mmol / g to 1.7 mmol / g, 0.01 mmol / g to 1.5 mmol / g, 0.05 mmol / g to 1.5 mmol / g, 0.1 mmol / g to 1.5 mmol / g, 0.3 mmol / g to 1.5 mmol / g, 0.5 mmol / g to 1.5 mmol / g, 0.7 mmol / g to 1.5 mmol / g, 1.0 mmol / g to 1.5 mmol / g, 0.01 mmol / g to 1.2 mmol / g, 0.05 mmol / g to 1.2 mmol / g, 0.1 mmol / g to 1.2 mmol / g, 0.3 mmol / g to 1.2 mmol / g, 0.5 mmol / g to 1.2 mmol / g, 0.7 mmol / g to 1.2 mmol / g, 1.0 mmol / g to 1.2 mmol / g, 0.01 mmol / g to 1 mmol / g, 0.05 mmol / g to 1 mmol / g, 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, but is not limited to, 7 mmol / g to 1 mmol / g.

[0059] In one embodiment of the present invention, the functional coating may have an expansion amount per unit mass of 2 mL / g to 8 mL / g in water. Specifically, the functional coating may have an expansion amount per unit mass of 2 mL / g to 8 mL / g, 2 mL / g to 7 mL / g, 2 mL / g to 6 mL / g, 2 mL / g to 5 mL / g, 3 mL / g to 8 mL / g, 3 mL / g to 7 mL / g, 3 mL / g to 6 mL / g, or 3 mL / g to 5 mL / g in water, but is not limited thereto. In addition, the functional coating causes swelling in a solvent other than water, and may have different expansion amounts per unit mass depending on the type of solvent.

[0060] In one embodiment of the present invention, the functional coating may occupy 1% to 15% of the pore volume of the core substrate without the functional coating. Specifically, the functional coating may occupy, but is not limited to, 1% to 15%, 1% to 12%, 1% to 10%, 1% to 7%, 1% to 5%, 1% to 3%, 3% to 15%, 3% to 12%, 3% to 10%, 3% to 7%, 3% to 5%, 5% to 15%, 5% to 12%, 5% to 10%, 5% to 7%, 7% to 15%, 7% to 12%, 7% to 10%, 10% to 15%, 10% to 12%, or 12% to 15% of the pore volume of the core substrate without the functional coating. In order to increase the efficiency of biological polymer synthesis, the flowability of the reaction solution in the composite solid support is very important. Therefore, it is necessary to appropriately control the coating amount and thickness of the functional coating in the composite solid support so that only the desired amount of pores in the core substrate are filled. Furthermore, the functional coating must be designed with swelling characteristics in mind. If the functional coating fills less than 1% of the pore volume of the core substrate, the desired amount of biological polymer synthesis may not occur. Furthermore, if the functional coating fills more than 15% of the pore volume, the reaction solution cannot flow smoothly, inevitably resulting in a large pressure drop.

[0061] In one embodiment of the present invention, the mass of the functional coating may be 5% to 200% of the mass of the core substrate without the functional coating. Specifically, the mass of the functional coating is 5% to 200%, 5% to 150%, 5% to 120%, 5% to 100%, 5% to 80%, 5% to 50%, 5% to 30%, 5% to 10%, 10% to 200%, 10% to 150%, 10% to 120%, 10% to 100%, 10% to 80%, 10% to 50%, 10% to 30%, 20% to 200%, 20% to 150%, 20% to 120%, 20% to 100%, 20% to 80%, 20% to 50%, 20% to 30%, It may be, but is not limited to, 30% to 200%, 30% to 150%, 30% to 120%, 30% to 100%, 30% to 80%, 30% to 50%, 50% to 200%, 50% to 150%, 50% to 120%, 50% to 100%, 50% to 80%, 80% to 200%, 80% to 150%, 80% to 120%, 80% to 100%, 100% to 200%, 100% to 150%, or 100% to 120%. If the functional coating is coated in an amount less than 5% of the mass of the core substrate, the desired amount of biological polymer synthesis may not occur, and if the coating is applied in an amount exceeding 200%, the reaction solution cannot flow smoothly, which inevitably leads to a large pressure drop.

[0062] In one embodiment of the present invention, the core substrate may have solvent resistance, heat resistance, or both. Unlike the functional coating, the core substrate does not exhibit swelling or solvent dissolution properties, and thus can serve as a support for a composite solid support. Furthermore, the core substrate is heat resistant, and thus can function as a support without change even when the synthesis of a biological polymer is carried out at high temperatures.

[0063] In one embodiment of the present invention, the component of the core substrate may include at least one selected from a polymer, a metal, and a ceramic. Specifically, the polymer may include at least one selected from, but is not limited to, high-density polyethylene (HDPE), low-density polyethylene (LDPE), linear low-density polyethylene (LLDPE), ultra-high molecular weight polyethylene (UHMWPE), polypropylene (PP), polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polyethylene naphthalate (PEN), polyamide (PA), polyvinyl chloride (PVC), polyvinyl difluoride (PVDF), polyacetal, polycarbonate (PC), polyimide (PI), polyetheretherketone (PEEK), polyethersulfone (PES), polyphenylene oxide (PPO), and polyphenylene sulfide (PPS). The metal may include at least one selected from among stainless steel, titanium, nickel, tantalum, zirconium, and alloys thereof, but is not limited thereto. The ceramic may include at least one selected from among fused silica, alumina, zirconia, silicon carbide, silicon nitride, boron nitride, and titanium diboride, but is not limited thereto.

[0064] In one embodiment of the present invention, the core substrate may have at least one shape selected from a one-dimensional shape, a two-dimensional shape, and a three-dimensional shape.

[0065] In one embodiment of the present invention, the one-dimensional shape may include at least one 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, and in this case, the core substrate is configured to have a length sufficient to enable biological polymer synthesis, and the composite solid support is loaded into a reactor in an entangled form, thereby being utilized in a biological polymer synthesis reaction.

[0066] In one embodiment of the present invention, the two-dimensional shape may include at least one selected from a spunbond non-woven fabric, a meltblown non-woven fabric, a needle-punched non-woven fabric, a hydroentangled non-woven fabric, a woven fabric, a knitted fabric, a porous membrane, a polymeric film, and a mesh. As a specific example, the two-dimensional shape of the core substrate may include a form in which a plurality of the one-dimensional shaped core substrates are randomly arranged. In addition, the two-dimensional shape may exhibit porosity by forming pores between the plurality of the one-dimensional shaped cores by the random arrangement.

[0067] In one embodiment of the present invention, the three-dimensional shape may include an open cell foam, a macropored sphere, or both.

[0068] In one embodiment of the present invention, the core substrate may have a length of one cross section of at least 1 mm, such that the functional coating can participate in the synthesis of the desired biological polymer. As a specific example, when the core substrate has a one-dimensional shape, a core substrate having a shape such as a fiber or the like can be implemented to serve as a support for a composite solid support through entanglement. In addition, when the core substrate has a two-dimensional or three-dimensional shape, it can serve as a support in itself, such as a non-woven fabric (two-dimensional) or a foam (three-dimensional). Therefore, the length of one cross section of the core substrate may be at least 1 mm, and theoretically, a one-dimensional core substrate with an infinite length can also serve as a support.

[0069] In one embodiment of the present invention, 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. In addition, the functional coating may be a polymer formed through a polymerization reaction of one or more main monomers as main components with an active monomer that provides a reaction site.

[0070] In one embodiment of the present invention, 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 by a polymerization of the one or more main monomers and the active monomer. Furthermore, in the heterogeneous composite solid support, the functional coating including a pulverized conventional solid support may be coated on the core substrate. The above conventional solid support may include at least one selected from, but is not limited to, polystyrene / divinylbenzene copolymer (PS / DVB), crosslinked polyethyleneglycol, poly-ε-lysine / sebacic acid, controlled pore glass, amino-polyacrylamide-resin fiber, cellulose, and hydroxylated polypropylene.

[0071] In one embodiment of the present invention, the one-dimensional core substrate may have a diameter of 10 μm to 100 μm. Specifically, the diameter 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 one-dimensional core substrate is less than 10 μm, it becomes too thin, causing the core substrate to clump together and resulting in poor mechanical properties. If it exceeds 100 μm, the amount of functional coating to be coated becomes too small, and the amount of biological polymer to be synthesized may become too small.

[0072] In one embodiment of the present invention, the two-dimensional core substrate may have a thickness of 10 μm to 10 mm. Specifically, the two-dimensional core substrate has a thickness of 10 μm to 10 mm, 50 μm to 10 mm, 100 μm to 10 mm, 150 μm to 10 mm, 200 μm to 10 mm, 250 μm to 10 mm, 300 μm to 10 mm, 350 μm to 10 mm, 400 μm to 10 mm, 10 μm to 5 mm, 50 μm to 5 mm, 100 μm to 5 mm, 150 μm to 5 mm, 200 μm to 5 mm, 250 μm to 5 mm, 300 μm to 5 mm, 350 μm to 5 mm, 400 μm to 5 mm, 10 μm to 3 mm, 50 μm to 3 mm, 100 μm to 3 mm, 150 μm to 3 mm, 200 μm to 3 mm, 250 μm to 3 mm, 300 μm to 3 mm, 350 μm to 3 mm, 400 μm to 3 mm, 10 μm to 1 mm, 50 μm to 1 mm, 100 μm to 1 mm, 150 μm to 1 mm, 200 μm to 1 mm, 250 μm to 1 mm, 300 μm to 1 mm, 350 μm to 1 mm, 400 μm to 1 mm, 10 μm to 0.5 mm, 50 μm to 0.5 mm, 100 μm to 0.5 mm, 150 μm to 0.5 mm, 200 μm to 0.5 mm, 250 μm to 0.5 mm, It may be, but is not limited to, 300 μm to 0.5 mm, 350 μm to 0.5 mm, or 400 μm to 0.5 mm. If the thickness of the two-dimensional core substrate is less than 10 μm, it is difficult to maintain mechanical properties, and if it exceeds 10 mm, there is a possibility that the polymerization reaction may not occur uniformly throughout the interior of the functional coating.

[0073] In one embodiment of the present invention, the porosity of the core substrate may be 50% to 95%. When the core substrate is one-dimensional, the core substrate having a shape such as fibers may form pores through entanglement, and when the core substrate is two-dimensional or three-dimensional, it may include pores inside, such as non-woven fabric or foam. In cases where the core substrate has pores as such, the porosity of the core substrate may be, but is not limited to, 50% to 95%, 60% to 95%, 70% to 95%, 80% to 95%, 90% to 95%, 50% to 90%, 60% to 90%, 70% to 90%, or 80% to 90%. Here, there is no particular limitation on the pore size of the core substrate. However, if the porosity is less than 50%, the pores may become blocked even after the functional coating swells, preventing the synthesis of biological polymers within the functional coating. Furthermore, if the porosity exceeds 95%, it may be difficult to maintain mechanical properties.

[0074] In one embodiment of the present invention, the thickness of the functional coating may be 0.1 μm to 1000 μm. The thickness of the functional coating may be selected to an appropriate thickness depending on the type of biological polymer desired and whether the core substrate has a one-dimensional, two-dimensional, or three-dimensional shape. Specifically, the thickness of the functional coating is 0.1 μm to 1000 μm, 0.5 μm to 1000 μm, 1 μm to 1000 μm, 5 μm to 1000 μm, 10 μm to 1000 μm, 50 μm to 1000 μm, 100 μm to 1000 μm, 0.1 μm to 500 μm, 0.5 μm to 500 μm, 1 μm to 500 μm, 5 μm to 500 μm, 10 μm to 500 μm, 50 μm to 500 μm, 100 μm to 500 μm, 0.1 μm to 200 μm, 0.5 μm to 200 μm, 1 μm to 200 μm, 5 μm to 200 μm, 10 μm to 200 μm, 50 μm to 200 μm, 100 μm to 200 μm, 0.1 μm to 100 μm, 0.5 μm to 100 μm, 1 μm to 100 μm, 5 μm to 100 μm, 10 μm to 100 μm, 50 μm to 100 μm, 0.1 μm to 50 μm, 0.5 μm to 50 μm, 1 μm to 50 μm, 5 μm to 50 μm, 10 μm to 50 μm, 0.1 μm to 20 μm, 0.5 μm to 20 μm, 1 μm to 20 μm, 5 μm to 20 μm or 10 μm to 20 The thickness of the functional coating may be, but is not limited to, μm. If the thickness of the functional coating is excessively small, such as less than 0.1 μm, the amount of the functional coating becomes small, resulting in a problem in that the amount of the biological polymer synthesized becomes very small.In addition, if the thickness of the functional coating is excessively large, such as exceeding 1000 μm, the functional coating itself may be manufactured through a non-uniform polymerization reaction, and a problem of reduced purity may occur during the biological polymer synthesis process.

[0075]

[0076] The third aspect of the present invention provides a method for producing a composite solid support for synthesizing biological polymers, comprising the steps of: preparing a functional coating solution by mixing one or more main monomers, an active monomer, an initiator, and a solvent; impregnating a core substrate having a length of 1 mm or more and a shape of one dimension or more into the functional coating solution; and polymerizing the functional coating solution applied to the core substrate.

[0077] In one embodiment of the present invention, the core substrate impregnated with the functional coating solution may have a hydrophilic surface treatment or may not be hydrophilic. Specifically, the surface hydrophilic treatment may be performed through a method selected from, but not limited to, a chemical method using a surfactant or an acidic solution, and a physical method including plasma treatment or UV irradiation.

[0078] In one embodiment of the present invention, the loading density may be controlled according to the length of the main monomer.

[0079] In one embodiment of the present invention, the main monomer may include a first monomer or a first monomer and a second monomer.

[0080] In one embodiment of the present invention, the functional coating solution may be prepared by mixing a first monomer, an active monomer, an initiator, and a solvent. In this case, the first monomer functions as a cross-linking agent, and by adjusting its length, the loading density provided by the added active monomer can be controlled. While a shorter first monomer may increase the loading density, steric hindrance may occur between the synthesized biological polymers. Therefore, an appropriate first monomer must be selected depending on the desired biological polymer.

[0081] In one embodiment of the present invention, the first monomer may include at least one selected from a bisacrylamide-based crosslinking agent, a methacryloyl group, an alkenyl group-substituted triazine, and an acrylate-based crosslinking agent, and specifically, the first monomer may include a polyethylene glycol-based agent, for example, polyethylene glycol diacrylate. Specifically, the first monomer may be used without limitation as long as it is a material that can react or combine with an active monomer for exposing a functional group internally and / or on the surface in a functional coating. As a non-limiting example, the first monomer may include at least one selected from polyethylene glycol diacrylate, N,N'-methylenebisacrylamide (MBA), ethylene glycol dimethacrylate (EGDMA), poly(ethylene glycol) dimethacrylate (PEGDMA), glycidyl methacrylate (GMA), divinyl sulfone (DVS), triethylene glycol divinyl ether (TEGDVE), and diallyl phthalate (DAP).

[0082] In one embodiment of the present invention, the active monomer is for providing a functional group to the interior and / or surface of the functional coating, and may include at least one selected from among acrylate-based, acrylamide-based, and methacrylamide-based materials. As non-limiting examples, the active monomers include N-(2-aminopropyl) methacrylamide hydrochloride, Aminoethyl Methacrylate Hydrochloride (AEMA.HCl), 2-Aminoethyl Methacrylate (2-AEMA), N-(3-Aminopropyl) methacrylamide hydrochloride (APMA), 4-Aminostyrene, N-(4-aminophenyl)methacrylamide, N,N-Dimethylaminoethyl Methacrylate (DMAEMA), N,N-Dimethylaminopropyl Acrylamide (DMAPAA), and It may include at least one selected from N-(2-aminoethyl)acrylamide hydrochloride.

[0083] In one embodiment of the present invention, the initiator may include a photoinitiator, a thermal initiator, or both. The above photoinitiator may be a UV initiator, and may be selected from the group consisting of 1-hydroxy-cyclohexyl-phenyl-ketone (Irgacure 907), 2-methyl-1[4-(methylthio)phenyl]-2-morpholinopropan-1-one (Irgacure 184C), 1-hydroxy-2-methyl-1-phenyl-propan-1-one (Darocur 1173), a mixed initiator of Irgacure 184C and benzophenone (Irgacure 500), a mixed initiator of Irgacure 184C and Irgacure 1173 (Irgacure 1000), 2-hydroxy-1-[4-(2-hydroxyethoxy)phenyl]-2-methyl-1-propanone (Irgacure 2959), and methylbenzoylformate (Darocur MBF), α,α-dimethoxy-alpha-phenylacetophenone (Irgacure 651), 2-benzyl-2-(dimethylamino)-1-[4-(4-morpholinyl)phenyl]-1-butanone (Irgacure 369), mixed initiator of Irgacure 369 and Irgacure 651 (Irgacure 1300), diphenyl(2,4,6-trimethylbenzoyl)-phosphine oxide (Darocur TPO), mixed initiator of Darocur TPO and Darocur 1173 (Darocur 4265), phosphine oxide, phenyl bis(2,4,6-trimethylbenzoyl) (Irgacure 819), mixed initiator of Irgacure 819 and Darocur 1173 (Irgacure 2005), It may include at least one selected from the group consisting of a mixed initiator of Irgacure 819 and Darocure 1173 (Irgacure 2010), a mixed initiator of Irgacure 819 and Darocure 1173 (Irgacure 2020), bis(eta 5-2,4,-cyclopentadien-1-yl)bis[2,6-difluoro-3-(1H-pyrrole-1-)phenyl]titanium (Irgacure 784), and a mixed initiator containing 2-hydroxy-2-methylpropiophenone and benzophenone (HSP 188).

[0084] In addition, the thermal initiator is an azo compound such as 2,2'-azobis(2-methylbutyronitrile), 2,2'-azobis(isobutyronitrile), 2,2'-azobis(2,4-dimethylvaleronitrile), 4,4-azobis(4-cyanovaleric acid), 1,1'-azobis(cyclohexanecarbonitrile), and 2,2'-azobis(4-methoxy-2,4-dimethylvaleronitrile); Tetramethylbutylperoxy neodecanoate (ex. Perocta ND, manufactured by NOF), bis(4-butylcyclohexyl)peroxydicarbonate (ex. Peroyl TCP, manufactured by NOF), di(2-ethylhexyl)peroxy carbonate, butylperoxy neodecanoate (ex. Perbutyl ND, manufactured by NOF), dipropyl peroxy dicarbonate (ex. Peroyl NPP, manufactured by NOF), diisopropyl peroxy dicarbonate (ex. Peroyl IPP, manufactured by NOF), diethoxyethyl peroxy dicarbonate (ex. Peroyl EEP, manufactured by NOF), diethoxyhexyl peroxy dicarbonate (ex. Peroyl OEP, manufactured by NOF), hexyl peroxy dicarbonate (ex. Perhexyl ND, manufactured by NOF), dimethyl Peroxybutyl peroxy dicarbonate (ex. Peroyl MBP, manufactured by NOF), bis(3-methoxy-3-methoxybutyl) peroxy dicarbonate (ex. Peroyl SOP, manufactured by NOF), dibutyl peroxy dicarbonate, dicetyl peroxy dicarbonate, dimyristyl peroxy dicarbonate, 1,1,3,3-tetramethylbutyl peroxypivalate, hexyl peroxy pivalate (ex. Perhexyl PV, manufactured by NOF), butyl peroxy pivalate (ex. Perbutyl, manufactured by NOF), trimethyl hexanoyl peroxide (ex. Peroyl 355, manufactured by NOF), dimethyl hydroxybutyl peroxyneodecanoate (ex. Luperox 610M75, manufactured by Atofina), Amyl peroxyneodecanoate (ex.Luperox 546M75, manufactured by Atofina), butyl peroxyneodecanoate (ex. Luperox 10M75, manufactured by Atofina), t-butylperoxy neoheptanoate, amylperoxy pivalate (ex. Luperox 546M75, manufactured by Alofina), t-butylperoxy pivalate, t-amyl peroxy-2-ethylhexanoate, lauryl peroxide, dilauroyl peroxide, didecanoyl peroxide, benzoyl peroxide, dibenzoyl peroxide, 2,2-bis(tert-butylperoxy)butane, 1,1-bis(tert-butylperoxy)cyclohexane, 2,5-bis(butylperoxy)-2,5-dimethylhexane, It may include at least one selected from the group consisting of peroxy compounds such as 2,5-bis(tert-butylperoxy)-1-methylethyl)benzene, 1,1-bis(tert-butylperoxy)-3,3,5-trimethylcyclohexane, tert-butyl hydroperoxide, tert-butyl peroxide, tert-butyl peroxybenzoate, tert-butylperoxy isopropyl carbonate, cumene hydroxyperoxide, dicumyl peroxide, lauroyl peroxide, and 2,4-pentanedione peroxide; tert-butyl peracetate, peracetic acid, and potassium persulfate.

[0085] In one embodiment of the present invention, the solvent is water (H2O), ethanol (C2H5OH), methanol (CH3OH), isopropanol (C3H8O), acetone (C3H6O), ethylene glycol (C2H6O2), propylene glycol (C3H8O2), acetonitrile (C2H3N), dimethylformamide (DMF, C3H7NO), dimethylsulfoxide (DMSO, C2H6OS), N,N-dimethylacetamide (DMA, C4H9NO), tetrahydrofuran (THF, C4H8O), dichloromethane (DCM, CH2C l2 ) and glycerin (C3H8O3).

[0086] In one embodiment of the present invention, the functional coating solution may be prepared by mixing a first monomer, a second monomer, an active monomer, an initiator, and a solvent. The first monomer functions as a crosslinking agent, and the second monomer functions as a binder. The second monomer may be any substance that can react or bind with the first monomer and the active monomer without limitation. More specifically, the second monomer may include at least one selected from 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).

[0087] In one embodiment of the present invention, the functional coating solution may further include a surfactant. Specifically, the surfactant may include at least one selected from sodium dodecyl sulfate (SDS), sodium lauryl ether sulfate (SLES), lauramine oxide, sodium myreth sulfate, cetyl trimethylammonium bromide (CTAB), Triton X-100, Tween 20, Tween 80, and decyl glucoside.

[0088] In one embodiment of the present invention, the functional coating solution may contain 1 to 25 parts by weight of the active monomer relative to 100 parts by weight of the main monomer. Specifically, the functional coating solution may contain 1 to 25 parts by weight of the active monomer and 0.1 to 10 parts by weight of the initiator relative to 100 parts by weight of the main monomer. More specifically, when the main monomer comprises two types, the main monomers may be divided into a first monomer and a second monomer. In this case, the functional coating solution may contain 0 to 50 parts by weight of the second monomer and 1 to 25 parts by weight of the active monomer relative to 100 parts by weight of the first monomer, or may contain 0 to 50 parts by weight of the second monomer, 1 to 25 parts by weight of the active monomer, 0.1 to 10 parts by weight of an initiator, and 0 to 25 parts by weight of the surfactant relative to 100 parts by weight of the first monomer. If the ratio of the active monomer for exposing the functional group is high, the amount of the synthesized biological polymer may increase, but the possibility of steric hindrance occurring between the biological polymers synthesized in the composite solid support is high. In addition, if the ratio of the first monomer (main monomer) is high, the durability of the functional coating may increase, but the amount of the synthesized biological polymer may decrease.

[0089] In one embodiment of the present invention, the functional coating solution may contain 10 to 40 parts by weight of the main monomer, 1 to 10 parts by weight of the active monomer, 0.05 to 5 parts by weight of the initiator, and 30 to 80 parts by weight of the solvent, relative to 100 parts by weight of the composite solid support. Specifically, when the main monomer is the first monomer, the functional coating solution may contain 10 to 40 parts by weight of the first monomer, 1 to 10 parts by weight of the active monomer, 0.05 to 5 parts by weight of the initiator, and 30 to 80 parts by weight of the solvent, relative to 100 parts by weight of the composite solid support. In the functional coating solution, the second monomer and the surfactant are not essential components and may be added depending on the synthesis of the desired biological polymer compound. In this case, the functional coating solution may contain 10 to 40 parts by weight of the first monomer, 0 to 10 parts by weight of the second monomer, 1 to 10 parts by weight of the active monomer, 0.05 to 5 parts by weight of the initiator, 0 to 10 parts by weight of the surfactant, and 30 to 80 parts by weight of the solvent, based on 100 parts by weight of the composite solid support.

[0090] In one embodiment of the present invention, the biological polymer may include, but is not limited to, one or more selected from peptides, oligonucleotides, and peptide nucleic acids (PNA).

[0091]

[0092] A fourth aspect of the present invention provides a method for synthesizing a biological polymer, comprising the steps of: coupling a first amino acid residue having a protecting group linked to a composite solid support according to the first aspect; removing the protecting group; and coupling a second amino acid residue having a protecting group linked to the N-terminus or C-terminus of the first amino acid, from which the protecting group is removed.

[0093] The method for synthesizing the above biological polymer provides a process by which a biological polymer desired in a functional coating can be synthesized by repeatedly performing the coupling step, the protecting group removal step, and the additional coupling step while changing the amino acid.

[0094] In one embodiment of the present invention, the method for synthesizing the biological polymer may further include a step of introducing a linker into the functional coating of the composite solid support before the step of coupling the first amino acid residue to which the protecting group is linked.

[0095] In one embodiment of the present invention, the method for synthesizing the biological polymer may further include a step of removing a protecting group from the introduced linker before the step of coupling the first amino acid residue to which the protecting group is linked.

[0096] In one embodiment of the present invention, the linker may include at least one selected from a Rink amide linker, a Wang linker, a 2-Chlorotrityl (CTC) linker, a Sieber linker, a BAL linker, a 4-sulfamylbutyryl linker, an HMBA (TFA stabilized) linker, an HMPA[4-(Hydroxymethyl)phenoxyacetylamidoethyl] linker, and an HMBA[4-(Hydroxymethyl)benzoylamidoethyl] linker.

[0097] In one embodiment of the present invention, the functional coating may have a functional group exposed internally and / or on the surface, and a linker may be bonded to the functional group. By adding additional monomer units (amino acid residues) to the monomer units (amino acid residues) connected via the functional coating (functional group)-linker, a biological polymer comprising multiple monomer units may be synthesized.

[0098] In one embodiment of the present invention, there is no limitation on the type of amino acid residue, and preferably, it may be selected from among alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, pyrrolysine, proline, glutamine, aruginine, serine, threonine, selenocysteine, valine, tryptophan, and tyrosine, but is not limited thereto.

[0099]

[0100] [Example]

[0101] Example 1. Preparation of polymer for functional coating and confirmation of swelling characteristics.

[0102] Example 1-1: Preparation of polymer for functional coating

[0103] To manufacture a polymer for functional coating, each reagent listed in Table 1 below was used, and each reagent was uniformly dissolved in a solvent by mixing and stirring. At this time, the components of the polymer for functional coating should satisfy each mass ratio (parts by weight) range, and the solvent can be used regardless as it will be dried later. In the experiment, each reagent and solvent (DW) were measured so that the sum of them was 100 g [the solvent can be used within 10 wt% to 90 wt% of the total (reagent + solvent)].

[0104] Classification Reagent Name Mass Part 1 Monomer Polyethylene Glycol Diacrylate 100 Second Monomer Methyl Acrylate 0~50 Active Monomer N-(2-aminopropyl) Methacrylamide Hydrochloride 1~25 Surfactant Sodium Dodecyl Sulfate 0~25

[0105]

[0106] Afterwards, 0.5 g of 2-hydroxy-2-methylpropiophenone, a curing reaction initiator, was added to the above solution and mixed uniformly to prepare a functional coating solution. The functional coating solution was uniformly applied to a petri dish and heated at 10 mJ / cm 2 1,000 mJ / cm 2 The UV energy was sufficiently irradiated to cure the polymer. After curing was complete, the reaction residue was removed using ethanol to obtain a functional coating polymer disk. The mass of the dried functional coating polymer was measured to be 45.7 g.

[0107] The manufactured functional coating polymer was rapidly frozen using liquid nitrogen and then ground using a mortar and pestle. Subsequently, a functional coating polymer in the form of particles with a particle size range of 100-200 mesh was obtained using a 100 or 200 mesh strainer (Example 1).

[0108] Example 1-2: Confirmation of the swelling characteristics of polymers for functional coatings in different solvents

[0109] The injection port of a 3 ml syringe was tightly sealed with glass fiber and the weight was measured. After that, approximately 200 mg of a functional coating polymer was filled and the initial volume was measured (initial volume V1). Then, 3 ml of a solvent to check the swelling characteristics was added to each syringe filled with the functional coating polymer and left for 30 minutes. After that, the solvent was slowly removed by pressing the pistol and the increased volume was measured (initial volume V2).

[0110] The volume change of the functional coating polymer due to the solvent was calculated (V2-V1), and the volume change value was divided by the mass of the functional coating polymer to calculate the swelling per unit mass. As a comparative example, the swelling per unit mass was calculated in the same way using commercially available PS / DVB polymer (crosslinked polystyrene / divinyl benzene copolymer) particles (Beadtech, Rink amide AM (MBHA) resin, 0.5 mmol / g, 100-200 mesh) (Table 2 and Fig. 1).

[0111] Swelling per unit mass by polymer solvent (mL / g) H2OMeOHEtOHEtOAcDMSODMFNMPTHFMCComparative Example 0.950.961.001.441.993.964.723.942.84Example 13.393.003.332.873.343.814.343.444.23

[0112]

[0113] Looking at the results in Table 2 and Figure 1 above, it was confirmed that Example 1 exhibited a greater degree of swelling than the comparative examples in solvents such as H2O, EtOH, MeOH, EtOAc, DMSO, and MC. In particular, swelling of more than twice was observed in eco-friendly solvents such as H2O, EtOH, MeOH, and EtOAc. This confirmed that Example 1 had superior swelling characteristics compared to the existing PS / DVB polymer under eco-friendly solvent conditions.

[0114]

[0115] Example 2. Measurement of loading density of polymer for functional coating

[0116] Example 2-1: Introduction of Fmoc-Gly-OH into a polymer for functional coating

[0117] In Example 1, 1 g of the functional coating polymer was added to 8 ml of DMF and swelled for 30 minutes, and the DMF was filtered to prepare a sample for measuring the loading density. Then, 3 mmol of Fmoc(fluorenyl methoxycarbonyl)-Gly(glycine)-OH, DIC, and Oxyma pure(Ethyl cyanohydroxyiminoacetate) were added and dissolved in 8 ml of clean DMF to prepare a reagent. The reagent was added to the swollen functional coating polymer sample and stirred at room temperature for 5 hours to react. Afterwards, the reaction solution was discarded and washed 7 times with 8 ml of DMF, 3 times with 8 ml of ethanol, and dried under reduced pressure for 12 hours.

[0118] Example 2-2: Loading density measurement

[0119] 8 mL of DMF was added to 1 g of the functional coating polymer with Fmoc-Gly-OH introduced, and swelling was performed for 30 minutes. 2 mL of a 20% piperidine solution was added to the sufficiently swollen functional coating polymer, and the reaction was performed at room temperature for 45 minutes. 1 mL of the supernatant was collected, diluted until the absorbance at 301 nm was approximately 1, and the loading density was analyzed. The loading density was measured to be 0.17 mmole / g, as calculated using Equation 1 below.

[0120] [Formula 1]

[0121] L = (A 301 Х V Х d) / (E c Х W Х m Х 1000)

[0122] Here, L = loading density (mmole / g)

[0123] A 301 = absorbance at 301 nm

[0124] V = volume of deprotection solution (piperidine solution)

[0125] d = dilution factor

[0126] E c= extinction coefficient

[0127] W = width of the cuvette

[0128] m = mass of the sample

[0129]

[0130] Example 3. Preparation of composite solid support

[0131] In order to manufacture a composite solid support, the functional coating solution (before curing) in 1-1) of the above Example 1 and the core substrate (polypropylene, spunbond nonwoven fabric, 40 g / m 2 ) 20 g was prepared. After sufficiently impregnating the core substrate in the functional coating solution and taking the core substrate out of the functional coating solution, it was cured in a UV curing chamber at 10 mJ / cm 2 - 1,000 mJ / cm 2 The ultraviolet energy was sufficiently irradiated.

[0132] After the curing process was completed, the coated core substrate was washed with ethanol to remove any remaining reaction residue and dried. A composite solid support was obtained in which a functional coating polymer was coated on the core substrate (solid support 30.4 g, 60.8 g / m). 2 ) and the appearance of the core substrate before coating is as shown in Fig. 2a, and the appearance of the composite solid support is as shown in Fig. 2b.

[0133]

[0134] Example 4. Surface treatment of core substrate

[0135] In Example 3, before impregnating the core substrate with the functional coating polymer curing reaction solution, the surface of the core substrate was hydrophilicized. The surface hydrophilicization was performed by impregnating the core substrate with a 2% SDS (sodium dodecyl sulfate) aqueous solution for 1 minute, taking it out, removing the excess SDS aqueous solution, and drying it at room temperature for 12 hours. Thereafter, a composite solid support was manufactured using the same method as in Example 3, and the difference between this and a composite solid support using a core substrate that was not subjected to the surface hydrophilicization treatment was confirmed. First, a fluorescent material, FITC (Fluorescein 5(6)-isothiocyanate), was chemically conjugated to each composite solid support, and the fluorescently tagged composite solid support was analyzed using a confocal laser scanning microscope (CLSM). In addition, SEM (scanning electron microscope) images were obtained to precisely observe the coated state.

[0136] As shown in a and b of Fig. 3, the composite solid support in which the core substrate was surface-treated was confirmed to have the FITC fluorescent material evenly distributed along the core substrate without agglomeration, indicating that the functional coating was evenly formed over the entire core substrate. On the other hand, the composite solid support using the core substrate that was not surface-treated was confirmed to have the functional coating formed in agglomerates in various places, as shown in a, b, c, and d of Fig. 4. This is a result that appears because the surface of the core substrate used in the experiment is hydrophobic and the polymer for functional coating is hydrophilic, meaning that if the surface treatment is appropriately performed depending on the type of core substrate and functional coating polymer, a more even functional coating can be formed.

[0137]

[0138] Example 5. Preparation of heterogeneous composite solid support

[0139] In addition to the functional coating polymer of Example 1, a heterogeneous composite solid support was prepared by coating a conventional PS / DVB polymer on a core substrate (hereinafter, the composite solid support prepared by curing the functional coating polymer of Example 3 is referred to as a homogeneous composite solid support, and the composite solid support coated with the conventional polymer is referred to as a heterogeneous composite solid support). Using the materials and method of Example 3, but without using an active monomer, the conventional PS / DVB polymer for bead-type solid-phase synthesis was pulverized to produce particles of 0.1 to 10 μm in size.

[0140] Looking at Fig. 5a, it can be confirmed that the composite solid support according to Example 3 has the form of a homogeneous composite solid support with a homogeneous surface. Fig. 5b shows a conventional PS / DVB polymer for bead-type solid-phase synthesis, and when this is coated on a core substrate, it can be confirmed that a heterogeneous composite solid support with a heterogeneous surface and an uneven core substrate is produced, as shown in Fig. 5c.

[0141]

[0142] Example 6. Coupling Kinetic Test Using Particulate Solid Support and Homogeneous / Heterogeneous Composite Solid Support

[0143] The homogeneous composite solid support subjected to the surface treatment of Example 4, the heterogeneous composite solid support of Example 5, and the particle-shaped solid support (PS / DVB polymer) were each placed in a reactor, and a coupling kinetics test was performed through the reaction steps of Table 3 below. First, 5 mL of DMF was added to each reactor and sufficiently swelled for 30 minutes. Then, 10 mL of a 20% piperidine solution was added, and the first deprotection was performed at 70°C for 10 minutes. After that, 1 mL of the supernatant was taken and placed in an Ep tube, and the coupling reaction was performed while varying the coupling reaction time (30 to 240 seconds). After washing, the second deprotection was performed, and 1 mL of the supernatant was placed in an Ep tube. The supernatants of the first and second deprotections were diluted by the same multiple and the UV spectrum was measured.

[0144] Step Reagent (solvent: DMF) Volume (mL) Temperature (℃) Time (min) Deprotection 120% piperidine 10 70 10 Washing DMF 10 (5 times) - Coupling Fmoc-Amino acid (0.2 M) 670 * DIC (0.5 M) 2 Oxyma pure (1.0 M) 1 Washing DMF 10 (5 times) - Deprotection 220% piperidine 10 70 10

[0145]

[0146] The yield of the coupling reaction (Coupling Yield (%)) was calculated using the absorbance of the second deprotected supernatant at 301 nm, taking the absorbance of the first deprotected supernatant as 100%. As shown in Fig. 6, the homogeneous and heterogeneous composite solid supports according to the examples showed a high coupling reaction yield even in a short coupling reaction time compared to the existing particle-type solid supports. In particular, the homogeneous composite solid support using an active monomer that participates in the curing reaction together showed a superior coupling reaction yield compared to the heterogeneous composite support, confirming that the homogeneous composite solid support is most suitable for use as a solid support for the synthesis of biological polymers.

[0147]

[0148] Example 7. Synthesis test using a batch-type automatic reactor

[0149] Acyl carrier protein (ACP) model peptide synthesis was performed using a CEM Liberty Blue automated reactor (ACP 65-74).

[0150] Sequence number 1: VQAAIDYING

[0151] 0.1 mmol of each of the particle-type solid support and the homogeneous composite solid support of Example 3 were added to a single batch reactor, and the reaction steps were repeated under the same conditions, only changing the type of amino acid in Table 4 below.

[0152] Step Reagent (solvent: DMF) Volume (mL) Temperature (℃) Time (min) Deprotection 20% piperidine, 0.1M Oxyma pure 15702 Washing DMF 10 (3 times) -- Coupling Fmoc-Amino acid (0.2M) 6704 DIC (0.5M) 2 Oxyma pure (1.0M) 1 Washing DMF 10 (2 times) --

[0153]

[0154] After the reaction, the particle-type solid support and the homogeneous composite solid support were thoroughly washed with EtOH and dried, respectively, and then subjected to a cleavage reaction for 2 hours. The cleavage reaction solution used here was a mixture of 95% trifluoroacetic acid (TFA), 2.5% triisopropylsilane (TIS), and 2.5% distilled water (DW). After cleavage, washing was performed by precipitation with cold ether and centrifugation. The ether washing was performed twice, and after sufficient drying by a reduced pressure drying method, the purity of the synthetic peptide was measured using high-performance liquid chromatography (HPLC) (Table 5).

[0155] Classification Purity (%) Yield (%) Homogeneous composite solid support 85.1 88.4 Particulate solid support 81.4 71.5

[0156]

[0157] Figures 7a and 7b are graphs confirming the purity and yield of ACP(65-74) synthesized using the homogeneous composite solid support of each example and the existing particle-type solid support. As shown in Table 5 and Figures 7a and 7b, the results of the synthetic test analysis using the existing commercial SPPS batch reactor (Liberty Blue, CEM) showed that the purity of the ACP(65-74) peptide synthesis was somewhat improved when the homogeneous composite solid support of the example was used, but the yield was significantly superior. These figures show that both the existing particle-type solid support and the homogeneous composite solid support are at a level that can be used industrially in a batch-type automatic reactor, and in particular, the homogeneous composite solid support is very excellent in terms of yield, indicating that it is more suitable for a batch type.

[0158]

[0159] Example 8. Synthesis test using a flow-type reactor

[0160] In order to compare the synthetic performance of a particulate solid support and a homogeneous composite solid support in a flow reactor, 0.5 mmol each of the particulate solid support and the homogeneous composite solid support of Example 3 were added to a 27 mL column, and then the reaction step was performed by changing the types of amino acids under the conditions shown in Table 6 below to synthesize the ACP (acyl carrier protein) model peptide (65-74) of SEQ ID NO: 1. Afterwards, the dry mass was measured to calculate the yield, and the purity of the synthesized peptide was measured using high-performance liquid chromatography (HPLC).

[0161] Reaction step Reagent (solvent: DMF) Volume (mL) Temperature (℃) Time (min) Flow rate (mL / min) Deprotection 20% piperidine, 0.1M Oxyma pure 2570 2100 Washing DMF 125 - Coupling Fmoc-Amino acid (0.2M) 1570 4 DIC (0.5M) 6 Oxyma pure (1.0M) 3 Washing DMF 20 - -

[0162]

[0163] As shown in Fig. 8, the synthetic performance in the flow reactor using the homogeneous composite solid support of the example was superior to that in the batch reactor of Example 7 in both synthetic purity and yield. In contrast, when the flow reactor was filled with the particulate solid support to proceed with the synthesis of ACP(65-74) peptide, a backpressure issue occurred during the synthesis, preventing the synthesis from proceeding properly. This is because the particulate solid support blocks the column of the flow reactor due to swelling, and the composite solid support in which the core substrate is coated with a functional coating is more suitable for use in the flow reactor because this problem is unlikely to occur.

[0164]

[0165] Example 9. Peptide synthesis test according to the first monomer type of the homogeneous composite solid support

[0166] In order to compare the loading density and peptide synthesis performance of homogeneous composite solid supports manufactured according to the length of the first monomer, homogeneous composite solid supports with different lengths of the first monomer were manufactured and experiments were conducted. Specifically, the homogeneous composite solid supports were manufactured using the same reagents and conditions as Table 1 of Example 1. In this case, the first monomer was polyethylene glycol diacrylate (PEG) C3H3O(OCH2CH2)nC3H3O2, and three types of PEG with different n (n=4, n=9, and n=14) were used.

[0167] Example 9-1: Measurement of loading density of homogeneous composite solid support

[0168] 8 ml of DMF was added to each homogeneous composite solid support and allowed to swell for 15 minutes. 2 ml of a 20% piperidine solution was added to the sufficiently swelled homogeneous composite solid support, and the mixture was allowed to react at room temperature for 45 minutes. 1 ml of the supernatant was then aliquoted. The supernatant was diluted until its absorbance was approximately 1, and the absorbance was measured. The reaction site spacing was calculated using Equation 1.

[0169] Example 9-2: Synthesis of peptides using the manufactured solid support for the synthesis of biological polymers

[0170] A homogeneous composite solid support using three types of PEGs (PEG-4, PEG-9, and PEG-14) with different lengths (1.13 g, 1.09 g, and 1.21 g, respectively) was placed in a 27 mL synthetic column, washed with 25 mL of DMF, and circulated at 70°C for 10 minutes at a rate of 100 ml / min. 1.63 g of Fmoc-Rink-Linker was dissolved in 15 mL of DMF in a conical tube, and then 6 mL of 0.5 M DIC and 3 mL of 1.0 M Oxyma pure were added. The solution in the conical tube was placed in the synthetic column and circulated at 70°C for 10 minutes. After this, the synthetic membrane was washed with 50 ml of DMF, and 25 ml of a deprotection solution (0.1 M Oxyma pure, 20% Piperidine in DMF) was circulated through the synthetic column at 50°C for 4 minutes, and then washed with 125 ml of DMF.

[0171] In another conical tube, 0.95 g of Fmoc-Gly-OH was dissolved in 15 ml of DMF, followed by the addition of 6 ml of 0.5 M DIC and 3 ml of 1.0 M Oxyma pure. After stirring for 5 minutes, the solution was placed in a column and circulated at 70°C for 10 minutes. The synthetic membrane was then washed with 50 ml of DMF.

[0172] A chain reaction for peptide synthesis was performed by sequentially adding the following compounds: Fmoc-Asn(Trt)-OH 1.91 g, Fmoc-Ile-OH 1.13 g, Fmoc-Tyr(tBu)-OH 1.47 g, Fmoc-Asp(OtBu)-OH 1.32, Fmoc-Ile-OH 1.13 g, Fmoc-Ala-OH 0.99 g, Fmoc-Ala-OH 0.99 g, Fmoc-Gln(Trt)-OH 1.95 g, and Fmoc-Val-OH 1.09 g.

[0173] 25 ml of a deprotection solution (0.1 M Oxyma pure, 20% Piperidine in DMF) was added to the above reaction column, circulated at 50°C for 4 minutes, and washed with 125 ml of DMF and 125 ml of ethanol. Afterwards, drying under reduced pressure was performed for 1 hour to obtain a homogeneous composite solid support on which the dried peptide was synthesized.

[0174] 10 ml of stripping solution (TFA: TIS: DW = 95:2.5:2.5) was added to three conical tubes, and the homogeneous composite solid support on which the peptide was synthesized was placed in each tube, and the reaction was performed at room temperature for 2 hours. After that, the solution in the tubes was collected, and 20 ml of cold diethyl ether was added to each to perform precipitation. The precipitated solution was centrifuged to obtain a solid, which was washed twice more using 20 ml of cold ether, and then sufficiently dried using reduced pressure drying to obtain the peptide of SEQ ID NO: 1.

[0175] The dry mass of ACP (65-74) synthesized using the above three types of synthetic membranes was measured to calculate the yield before processing, and high performance liquid chromatography (HPLC) and MALDI-TOF / TOF were performed to measure the purity, and the results are shown in Table 7.

[0176] PEG type Functional coating Loading density (mmol / g) Purity (%) Yield (%) n = 40.25 85.284.4 n = 90.16 98.294.3 n = 140.12 99.495.6

[0177]

[0178] Through Table 7, it was confirmed that the larger the spacing between reaction sites of the solid support for biological polymer synthesis by the PEG, the higher the yield and purity of the synthesized peptide.

[0179] From the above results, it was confirmed that as the length of the first monomer PEG increases, the loading density of the functional coating decreases, but the purity and yield of peptide synthesis increase. That is, if the length of the first monomer is long, the distance between the reaction sites in the functional coating can be increased, so the purity and yield can be increased. However, if the length of the first monomer is excessively long, the loading density may decrease, which may make it difficult to use when a large amount of peptide synthesis is required. In addition, if the length of the first monomer is short, the loading density may increase, but if the length is very short, the swelling characteristic may become very large, which may cause a problem of reduced durability. Therefore, an appropriate selection of the first monomer length is necessary depending on the target synthesis target, and conversely, the composite solid support of the example can be interpreted as being applicable to the synthesis of various biological polymers because the loading density and purity / yield can be controlled by controlling the length of the first monomer.

[0180]

[0181] Example 10. Synthesis test by flow rate in a flow reactor

[0182] In Example 3, the yield and purity of peptide synthesis according to the flow rate in a flow reactor were confirmed using a composite solid support. ACP(65-74) was synthesized using the same method as in Example 9, and the results of the purity and yield of the synthesized ACP(65-74) peptide before processing are shown in Table 8 and Figures 9a to 9c.

[0183] Flow rate (mL / min) Purity (%) Yield (%) 10084.985.320088.390.530091.195.7

[0184]

[0185] Figures 9a to 9c show HPLC graphs for synthesized ACP (65-74) according to flow rate in a flow reactor using a composite solid support of the example.

[0186] As shown in the drawing and Table 8, it was confirmed that the purity and yield of the peptide increased as the flow rate of the column increased. This shows that the composite solid support of the example can be suitably used in a flow reactor that must synthesize various biological polymers, as it exhibits a purity and yield of 80% or more even at various flow rates.

[0187]

[0188] Example 11. Comparison of peptide synthesis purity according to surface treatment of core substrate in the preparation of homogeneous and heterogeneous composite solid supports.

[0189] As in Example 4, the purity of peptide synthesis of SEQ ID NO: 2 was compared when using a solid support including a surface-untreated core substrate and a surface-treated core substrate.

[0190] Sequence number 2: WFTTLISTIM

[0191] First, in Example 3, various types of surface hydrophilic treatments were performed before impregnating the core substrate with the functional coating solution. The types of hydrophilic treatments are shown in Table 9.

[0192] Surface hydrophilic treatment method Chemical method Immerse in surfactant aqueous solution such as SDS (sodium dodecyl sulfate), SDBS (sodium dodecylbenzenesulfonate), Triton-X100, etc., then dry Acidic solution Immerse in acid aqueous solution such as hydrochloric acid, acetic acid, acetic anhydride, phosphoric acid for a certain period of time, then rinse with a large amount of water and dry Physical method Plasma Surface treatment is performed using a plasma generator UV Surface treatment is performed using a UV light source

[0193]

[0194] Each manufactured composite solid support was introduced into the column, washed with 25 ml of DMF, and then circulated with 25 ml of DMF for 10 minutes. Then, 6 equivalents of Fmoc-Rink amide MBHA Linker-OH, DIC, and Oxyma pure were dissolved in 25 ml of DMF and circulated at 70°C for 10 minutes at a flow rate of 100 ml. Then, the column was washed twice with 25 ml of DMF, and 25 ml of a deprotection solution (0.1 M, Oxyma pure, 20% piperidine in DMF) was circulated at 50°C for 4 minutes. Finally, the composite solid support with the functional coating formed was washed twice with 50 ml of DMF and once with 25 ml of DMF. Afterwards, instead of Fmoc-Rink amide MBHA Linker-OH, the following compounds were used, and the steps from the initial washing step using DMF to the washing step of the composite solid support on which the functional coating was formed using DMF were repeated: [Fmoc-Met-OH, Fmoc-Ile-OH, Fmoc-Thr(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ile-OH, Fmoc-Leu-OH, Fmoc-Thr(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Phe-OH, Fmoc-Trp(Boc)-OH]

[0195] Finally, a composite solid support with formed peptides was obtained and washed three times with 25 ml of ethanol. After drying, a cleavage reaction was performed for 2 hours. The cleavage reaction solution used here was a mixture of 95% TFA, 2.5% TSI, and 2.5% DW. After the cleavage reaction, ether washing was performed by precipitating with cold ether and centrifuging. The ether washing was performed twice, and then the product was sufficiently dried under reduced pressure.

[0196] Hydrophilic treatment purity (%) Untreated 58.16 SDS 62.41 Triton X-100 65.61 Acetic acid 67.86 Hydrochloric acid 65.17 Plasma 65.59 UV 62.26

[0197]

[0198] Table 10 above shows the purity of peptide synthesis using composite solid supports manufactured differently depending on the hydrophilic treatment method of the core substrate. First, it was confirmed that the purity of the core substrate without hydrophilic treatment was lower than that of the core substrate with hydrophilic treatment. In addition, there was no significant difference between the chemical and physical methods in terms of the hydrophilic treatment method. Therefore, in the synthesis of biological polymers using the composite solid support of the example, it was confirmed that the hydrophilic treatment of the core substrate during the manufacturing process was more advantageous for synthesis, and that the hydrophilic treatment used here can be used regardless of the physical or chemical method.

[0199]

[0200] Example 12. Preparation of composite solid supports according to functional coating content and peptide synthesis test.

[0201] Example 12-1: Control of functional coating content

[0202] A composite solid support was manufactured as in Example 3, but the amount of functional coating applied to the core substrate was controlled. Specifically, the core substrate was sufficiently impregnated with the functional coating solution, and after the core substrate was removed from the coating solution, it was passed between rollers at a constant interval to remove a certain amount of the functional coating solution contained in the solid support. The content of the functional coating applied to the solid support was controlled according to the mass and thickness shown in Table 11 below.

[0203] Example 12-2: Porosity Measurement and Peptide Synthesis

[0204] Solid support without functional coating (polypropylene, spunbond nonwoven, density 0.89 g / cm 3 ) was cut into 10*10 cm pieces, and the mass and thickness were measured, and the porosity was calculated (CTR 0). Here, the thickness was measured at 10 points at 0.5 cm intervals on the specimen, and the average was used. The thickness when a constant pressure was applied using a ratchet stop was used. In addition, the mass and thickness of the composite solid support manufactured previously were measured in the same manner, and the porosity reduction was calculated (CTR 1~6).

[0205]

[0206] CTR 0CTR 1CTR 2CTR 3CTR 4CTR 5CTR 6Mass (mg)295315368420537652801Thickness (mm)0.2580.2550.2610.2590.2620.2640.272Area (mm) 2 )1000010000100001000010000100001000010000Density (g / cm) 3 )0.890.890.890.890.890.890.890.89Porosity(%)87868482777267△Porosity0.00-1.03-2.99-5.37-10.18-14.90-20.24

[0207]

[0208] △In order to test the peptide synthesis of the composite solid support according to the porosity, ACP 65-74 peptide was synthesized as in Example 8, and its purity was confirmed by HPLC. The synthesis results are as shown in Table 12 and Fig. 10, and it was confirmed that as the thickness and mass of the functional coating increased, the porosity decreased, but the peptide synthesis purity gradually increased. In particular, the best synthesis purity was observed at CTR 2, and the desired level of synthesis purity was shown up to CTR 5. However, when the coating thickness became too thick, the synthesis purity was lowered, which is interpreted as the result of the porosity being too low and the reaction solution not flowing smoothly.

[0209] CTR 0CTR 1CTR 2CTR 3CTR 4CTR 5CTR 6△Porosity 0.00-1.03-2.99-5.37-10.18-14.90-20.24Purity (%) 0.0095.596.295.893.787.257.6

[0210]

[0211] Example 13. Synthesis of various biological polymers using composite solid supports.

[0212] To verify that the solid-phase support synthesis technology using a composite solid support is not limited to peptide synthesis and can synthesize various types of sequence-defined biopolymers such as peptide nucleic acid (PNA), peptoids, and oligourea foldamers, the following experiments were conducted.

[0213] Example 13-1: Synthesis of PNA-peptide using a flow reactor

[0214] First, 1.5 g of the composite solid support was placed in a reaction column, washed with 25 ml of DMF, and circulated at 70°C for 10 minutes at a rate of 100 ml / min. 1.62 g of Fmoc-Rink-Linker was dissolved in 15 ml of DMF in a conical tube, and then 3 ml of 1 M DIC and 6 ml of 0.5 M Oxyma pure were added. The solution in the tube was transferred to the column, circulated at 70°C for 10 minutes, and then the synthetic membrane was washed with 50 ml of DMF.

[0215] Afterwards, 25 ml of deprotection solution (0.1 M Oxyma pure, 20% piperidine in DMF) was circulated through the column at 50°C for 4 minutes, and then washed with 125 ml of DMF. 178 mg of Fmoc-Gly-OH was dissolved in 15 ml of DMF in a conical tube, and then 3 ml of 1.0 M DIC and 6 ml of 0.5 M Oxyma pure were added. After stirring for 5 minutes, the solution was transferred to the column, circulated at 70°C for 10 minutes, and then the synthetic membrane was washed with 50 ml of DMF.

[0216] Afterwards, the chain reaction for PNA-peptide synthesis was performed by sequentially replacing Fmoc-Gly-OH with the following compounds: [Fmoc-G(Bhoc)-OH 445 mg, Fmoc-T-aeg-OH 304 mg, Fmoc-C(Bhoc)-OH 421 mg, and Fmoc-A(Bhoc)-aeg-OH 435 mg]. Afterwards, 25 ml of deprotection solution was added to the column, which was circulated at 50°C for 4 minutes, and the synthetic membrane was washed with 125 ml of mixed solvent and 125 ml of ethanol. The PNA-peptide of the following SEQ ID NO: 3 was prepared by drying under reduced pressure for 1 hour.

[0217] Sequence number 3: ACTG-Gly

[0218] After adding 15 ml of stripping solution (TFA: Thioanisole: DW: PhOH: EDT = 82.5: 5: 5: 5: 2.5) to a conical tube, NH2-ACTG-Gly-complex solid support was added and reacted at room temperature for 3 hours. Only the solution in the tube was collected and added to 120 ml of cold diethyl ether to carry out precipitation. The precipitated solution was centrifuged to obtain a solid, washed twice more by adding 120 ml of cold ether, and then sufficiently dried using reduced pressure drying.

[0219] Example 13-2: Synthesis of PNA-peptide using conventional solid-phase synthesis method

[0220] First, 0.25 g of Fmoc-Rink amide MBHA resin was added to a reactor filled with a particulate solid support, followed by the addition of 5 ml of DMF and stirring for 10 minutes. Thereafter, 5 ml of a deprotection solution was added, and the mixture was mixed using nitrogen at room temperature for 5 minutes. This process was repeated once more. The reactor was washed three times with 10 ml of a mixed solution (DMF:DCM = 1:1). 1.5 ml of 0.2 M Fmoc-Gly-OH, 0.6 ml of 0.5 M DIC, 0.3 ml of 1.0 M Oxyma pure, 3 ml of 1.0 M DIPEA, and 0.3 ml of 1.0 M 2,6-Lutidine were added to a conical tube. After mixing the solutions in the tube, they were transferred to the reactor and reacted at room temperature for 30 minutes. This process was repeated once more. Next, it was washed six times using 10 ml of mixed solution.

[0221] Afterwards, the chain reaction for PNA-peptide synthesis was performed by sequentially replacing Fmoc-Gly-OH with the following compounds: [Fmoc-G(Bhoc)-OH, Fmoc-T-aeg-OH, Fmoc-C(Bhoc)-OH, Fmoc-A(Bhoc)-aeg-OH]. After that, 5 ml of deprotection solution was added and mixed using nitrogen at room temperature for 5 minutes. This was repeated once more. The reactor was washed three times with 10 ml of the above mixed solution 2, thereby producing the PNA of the above sequence number 3.

[0222] Afterwards, 5 ml of stripping solution (TFA: TIS: DW: PhOH: EDT = 82.5: 5: 5: 5: 2.5) was added to the conical tube. The synthesized NH2-ACTG-Gly-complex solid support was added and reacted at room temperature for 3 hours. Only the solution in the tube was collected, and TFA was removed using nitrogen. Precipitation was performed by pouring into 15 ml of cold ether. The precipitated solution was centrifuged to obtain a solid, which was washed twice more using 15 ml of cold ether, and then sufficiently dried using reduced pressure drying.

[0223] The dry mass of the PNA-peptide synthesized using the above composite solid support and the existing particle-type solid support was measured to calculate the yield before processing, and high performance liquid chromatography (HPLC) and MALDI-TOF / TOF were performed to measure the purity, and the results are shown in Table 13.

[0224] Solid support purity (%) yield (%, with TFA salt) homogeneous composite solid support 90.0 105.3 particulate solid support 86.8 103.0

[0225]

[0226] As shown in Table 13, the yield and purity of the PNA-peptide synthesized using the composite solid support of the example were improved compared to the PNA-peptide synthesized using the existing particle-type solid support. From this, it was confirmed that the composite solid support can be used for the synthesis of various types of biological polymers, such as peptides such as ACP(65-74), as well as peptide nucleic acids.

[0227] The above description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner.

[0228] The scope of the present invention is indicated by the claims described below rather than the detailed description above, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A core substrate (Structure core) having a length of 1 mm or more and a shape of one dimension or more; and A composite solid support comprising a functional coating positioned on the core substrate, In the above functional coating, a biological polymer is synthesized. Composite solid supports for the synthesis of biological polymers.

2. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the functional coating has the property of swelling in a solvent.

3. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the functional coating comprises a functional group on the surface, inside or both.

4. In paragraph 1, A composite solid support for the synthesis of biological polymers, wherein the loading density of the functional coating is 0.01 mmol / g to 2 mmol / g.

5. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the functional coating occupies 1% to 15% of the pore volume of the core substrate without the functional coating.

6. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the mass of the functional coating is 5% to 200% of the mass of the core substrate without the functional coating.

7. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the core substrate has the properties of solvent resistant, thermal resistant, or both.

8. In paragraph 1, A composite solid support for biological polymer synthesis, wherein the core substrate component comprises at least one selected from polymers, metals, and ceramics.

9. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the core substrate has at least one shape selected from a one-dimensional shape, a two-dimensional shape, and a three-dimensional shape.

10. In paragraph 9, A composite solid support for synthesizing biological polymers, wherein the one-dimensional shape includes at least one selected from staple fibers, filament fibers, and rods.

11. In paragraph 9, A composite solid support for synthesizing biological polymers, wherein the two-dimensional shape comprises at least one selected from a spunbond non-woven fabric, a meltblown non-woven fabric, a needle-punched non-woven fabric, a hydroentangled non-woven fabric, a woven fabric, a knitted fabric, a porous membrane, a polymeric film, and a mesh.

12. In paragraph 9, A composite solid support for synthesizing biological polymers, wherein the three-dimensional shape comprises an open cell foam, a macropored sphere, or both.

13. In paragraph 1, A composite solid support for synthesizing biological polymers, wherein the functional coating comprises a polymer of one or more main monomers and an active monomer.

14. In paragraph 2, A composite solid support for the synthesis of biological polymers, wherein the functional coating has a swelling capacity per unit mass of 2 mL / g to 8 mL / g in water. 15.1 A step of preparing a functional coating solution by mixing at least one main monomer, an active monomer, an initiator, and a solvent. A step of impregnating a core substrate having a length of 1 mm or longer and a shape of one dimension or longer into the functional coating solution; and A step of polymerizing the functional coating solution applied to the above core substrate. A method for producing a composite solid support for synthesizing biological polymers, comprising:

16. In paragraph 15, A method for producing a composite solid support for synthesizing biological polymers, wherein the core substrate impregnated with the functional coating solution has a hydrophilic surface.

17. In paragraph 15, A method for producing a composite solid support for biological polymer synthesis, wherein the loading density is controlled according to the length of the main monomer.

18. A step of coupling a first amino acid residue linked to a protecting group to a composite solid support according to paragraph 1; a step of removing the above protector; and A step of removing the above protecting group and coupling a second amino acid residue to which a protecting group is linked to the N-terminus or C-terminus of the first amino acid to which the above protecting group is coupled. A method for synthesizing a biological polymer, comprising:

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