Solid phase synthesis support, its preparation and use

Abstract

This solid-phase synthesis support, its manufacturing method, and use involve suspension polymerization of a diene crosslinker, which has a reactivity ratio similar to that of styrene and does not have two vinyls on the same benzene ring, as a crosslinking monomer to produce a porous resin. The porous resin is then functionalized to obtain a porous resin with amino or hydroxy functional groups. Compared to conventional manufacturing methods, the crosslinker's reactivity ratio similar to that of styrene improves the uniformity of the chemical structure within the resin, favoring the formation of uniformly distributed active sites and channels, which is advantageous for improving reaction efficiency and reducing mass transfer resistance. Using this support as a solid-phase synthesis support to produce oligonucleotides can improve the yield and purity of the oligonucleotides.

Description

[Technical field]

[0001] The present invention relates to a solid-phase synthesis support, its preparation method and use, which is applicable to the solid-phase synthesis of oligonucleotides, and belongs to the field of preparation of functional polymer materials. [Background technology]

[0002] As one of the effective methods of precision medicine, more and more oligonucleotide drugs have been approved and launched. Oligonucleotide drugs are often synthesized by chemical methods, the most common being the solid-phase phosphoramidite triester method, the specific method being to fill a solid-phase synthesis support into a reaction column, and then rapidly flow the solution containing the reactants through the reaction column at a certain pressure to complete the reaction.

[0003] In the early development of oligonucleotide solid-phase synthesis technology, the commonly used solid-phase synthesis supports are inorganic particles such as pore-controllable glass microspheres (CPG) and modified silicone, but their disadvantages are also obvious, namely, due to low substitution degree (loading) (generally less than 100 μmmol / g), the production volume of one batch of oligonucleotide is limited, the utilization rate of the equipment is low, and the production cost is high. In order to increase the substitution degree of the support, the invention patent WO2006029023 jointly applied for by Nitto Denko and Ionis uses styrene, 4-acetoxystyrene, and divinylbenzene as polymerization monomers, and isooctane and 2-ethylhexanol as pore-forming agents to prepare an organic polymer to form a solid-phase synthesis support, and the substitution degree of this support can reach 100 to 350 μmmol / g. In order to further improve the yield and purity of oligonucleotides, the invention patent US8653152 applied for by Nitto Denko uses styrene, (meth)acrylonitrile, 4-acetoxystyrene, and divinylbenzene as polymerization monomers, and uses isooctane and 2-ethylhexanol as pore-forming agents to produce a solid-phase synthesis support. The addition of (meth)acrylonitrile suppresses the swelling fluctuation of the solid-phase synthesis support in different solvents, improving the synthesis efficiency.

[0004] Both of the above two types of polymer carriers use divinylbenzene as a crosslinking agent. In use, divinylbenzene has two main problems: it is not very pure and there are three isomers with different properties. This causes a large difference in the performance of resins produced by different manufacturers and with different qualities of divinylbenzene. Currently, the purity of general industrial divinylbenzene is 40-60%, and in addition to divinylbenzene, it also contains a certain amount of ethylstyrene, and some also contain a small amount of diethylbenzene.

[0005] In copolymerization reactions with other monomers, the three isomers of divinylbenzene show different reaction activities, which have a great influence on the structure and properties of the copolymer. He Binglin et al. (Reactive Polymer, 12 (1990), 269-275) discovered in their study of the kinetics of parastyrene-divinylbenzene copolymerization that paradivinylbenzene and metadiethylene have a fundamentally faster conversion rate than styrene during the copolymerization process. This results in copolymers formed in the early stages of the polymerization reaction having a high degree of crosslinking, while those formed in the later stages of polymerization have a low degree of crosslinking, and the chemical structure inside the resin is heterogeneous. Due to this heterogeneity in chemical structure, macroporous adsorption resins with styrene-divinylbenzene as the skeleton have problems such as various degrees of fracture during use, long time to reach adsorption equilibrium, easy tailing of desorption after adsorption, and incomplete regeneration.

[0006] Diene crosslinkers such as 1,2-bis(4-vinylphenyl)ethane, which do not have two vinyl groups on the same benzene ring, have a reactivity similar to that of styrene. Sundell et al. (Journal of Polymer Science Part A: Polymer Chemistry, 31 (1993), 2305-2311) produced strong acid resins using such monomers as crosslinkers, and experimental results showed that these resins have higher mechanical strength than strong acid resins based on styrene-divinylbenzene copolymers. In "Study on the synthesis of 1,2-bis(p-vinylphenyl)ethane polymers and their adsorption performance," ultra-high crosslinked polystyrene adsorption resins were produced using 2-bis(p-vinylphenyl)ethane and divinylbenzene as crosslinkers, and experimental results showed that adsorption resins using 1,2-bis(p-vinylphenyl)ethane as a crosslinker did not have a tailing phenomenon during use and were easy to regenerate.

[0007] Oligonucleotide drugs have the advantages of high therapeutic efficiency, low drug toxicity, strong specificity, wide application range, etc., and are therefore effective in the treatment of diseases such as antiviral, high cholesterol, gene expression editing, visual blindness and hepatic vein obstruction. The demand for oligonucleotide drugs is increasing year by year. In addition, traditional oligonucleotide solid-phase synthesis supports have problems such as non-uniform internal structure and large mass transfer resistance, which leads to low oligonucleotide synthesis efficiency and high production costs. Therefore, in order to meet the demand of the oligonucleotide drug market, it is necessary to develop a solid-phase synthesis support that can synthesize oligonucleotide drugs on a large scale, at low cost and with high efficiency. Summary of the Invention [Problem to be solved by the invention]

[0008] In order to produce oligonucleotides on a large scale at low cost and to overcome the disadvantages of conventional oligonucleotide solid-phase synthesis supports, such as non-uniform internal structure and high mass transfer resistance, the present invention provides a solid-phase synthesis support, its preparation method and use.

[0009] In the present invention, the use of a diene compound in which two vinyls are not on the same benzene ring as a crosslinking monomer has three main effects. First, such a monomer has a reactivity ratio close to that of styrene, which is advantageous for improving the uniformity of the chemical structure inside the resin and forming uniformly distributed active sites. Second, the uniformity of the chemical structure inside the resin is improved, which is advantageous for forming uniform channels inside the resin, thereby reducing the mass transfer resistance. Third, compared with divinylbenzene, such a crosslinking agent has flexible molecular chains, which can improve the swelling of the resin in different organic solvents. [Means for solving the problem]

[0010] The present invention discloses a solid phase synthesis support having a polymer backbone and functional group structure represented by the following formula: [ka] (where R 1 =-(CH 2 ) n -(n is an integer of 0 to 3), or -O-(CH 2 ) m -O- (m is an integer from 1 to 4), and R 2 = -OH or -NH 2 It is.)

[0011] In some embodiments, the solid phase synthesis support is a copolymer whose backbone has repeating structural units of formula (I), formula (II), formula (III), or formula (IV). [ka] (where R 3 is -H, -CN, or -CH 2 -CN and R 4 is -H or -CH 3 and R 5 -CN, -CH 2 -CN, -COOCH 3 or -CONH 2 It is.) [ka] (where R 6 Ha-(CH 2 )x-(x is an integer of 0 to 3) or -O-(CH 2 ) y -O- (wherein y is an integer of 1 to 4). [ka] (R 7 -H, CH 3 (CH 2 ) z -, CH 3 (CH 2 ) z O-(z is an integer of 0 to 4), (CH 3 ) 2 CH-, (CH 3 ) 2 CH(CH 2 )-, (CH 3 ) 2 CH(CH 2 ) 2 -, (CH 3 ) 3 C-, CH 3 CH 2 CH(CH 3 )-, CH 3 CH 2 C(CH 3 ) 2 - or CH 3 CH 2 CH 2 CH(CH 3 )-It is. [ka] (R 8 -OH, -CH 2 OH, -NH 2 , -CH 2 NH 2 , -CH 2 OOC-C 6 H 4 -OH, -CH 2 OOCCH2 -C 6 H 4 -OH, -(CH 2 ) 4 OOC-C 6 H 4 -OH, -(CH 2 ) 4 OOCCH 2 -C 6 H 4 -O H、- COO-C 6 H 4 -OH or -CH 2 COO-C 6 H 4 -OH.

[0012] In some embodiments, the hydroxy or amino content of the carrier ranges from 100 to 1000 μm ol / g, preferably 400 to 700 μm ol / g.

[0013] In some embodiments, the carrier has a particle size in the range of 35-200 μm, preferably 50-100 μm.

[0014] In some embodiments, the carrier has an average pore size of 10 to 200 nm, preferably 40 to 100 nm.

[0015] The present invention relates to The present invention further discloses a method for producing the above solid phase synthesis support, the method comprising the steps of respectively preparing an aqueous phase consisting of water, a dispersant and an inorganic salt, and an oil phase consisting of a cross-linking monomer, a monovinyl compound, a functional monomer, a modified monomer, a pore-forming agent and an initiator, the cross-linking monomer, the monovinyl compound, the functional monomer and the modified monomer being involved in a polymerization reaction and collectively referred to as monomers, and adding the oil phase to the aqueous phase, stirring and heating to cause a reaction, removing the pore-forming agent after the reaction is completed to obtain a porous polymer resin, and further reacting the porous polymer resin to obtain a solid phase synthesis support containing hydroxy or amino as a functional group.

[0016] In some embodiments, more specifically, the crosslinking monomer is a diene crosslinker in which the two vinyls are not on the same benzene ring, including 4,4'-divinylbiphenyl, bis(4-vinylphenyl)methane, 1,2-bis(4-vinylphenyl)ethane, 1,3-bis(4-vinylphenyl)propane, di(4'-vinylphenoxy)methane, 1,2-bis(4'-vinylphenoxy)ethane, 1,3-bis(4'-vinylphenoxy)propane, and 1,4-bis(4'-vinylphenoxy)butane. A preferred crosslinking monomer is one or more of 1,2-bis(4-vinylphenyl)ethane.

[0017] In some embodiments, the monovinyl compound is an aromatic monovinyl compound, including styrene and its benzene ring substituted derivatives, styrenes having an alkyl substituent with 1 to 5 carbon atoms, such as methylstyrene, ethylstyrene, normal propylstyrene, isopropylstyrene, normal butylstyrene, isobutylstyrene, s-butylstyrene, t-butylstyrene, n-pentylstyrene, isopentylstyrene, s-pentylstyrene, or t-pentylstyrene; or styrenes having an alkoxy substituent with 1 to 5 carbon atoms, such as methoxystyrene, ethoxystyrene, propoxystyrene, butoxystyrene, or pentyloxystyrene. The preferred monovinyl compound is styrene.

[0018] In some embodiments, the functional monomer has a double bond capable of radical polymerization and a hydroxy, amino, halogenated group, or other group that can be converted to hydroxy or amino by reaction. In the process of synthesizing an oligonucleotide, the reactive hydroxy or amino becomes an active site for linking the oligonucleotide, and may be amino, aminoalkyl, hydroxy, hydroxyalkyl, etc. Preferably, it may be primary amino, aminomethyl, hydroxy, hydroxymethyl, etc. Hydroxystyrene and its derivatives, such as 4-hydroxystyrene, hydroxyalkylstyrene and its derivatives, such as 4-hydroxymethylstyrene; acyloxystyrene and its derivatives, such as 4-acetoxystyrene, benzoyloxystyrene, etc.; aminostyrene and its derivatives, such as 4-aminostyrene, aminoalkylstyrene and its derivatives, such as 4-aminomethylstyrene, etc.; haloalkylstyrene monomers, such as 4-(4-bromobutyl)styrene, p-chloromethylstyrene, etc.; 4-vinylphenyl ester monomers, such as methyl 4-vinylbenzoate, ethyl 4-vinylbenzoate, etc., but are not limited thereto.

[0019] In some embodiments, the functional monomers include those that contain a hydroxyl or amino protecting group, and the protecting group of these monomers can be directly cleaved to form amino or hydroxyl. For example, acyloxystyrene can be converted to hydroxyl by alkaline or acid hydrolysis to become an active site for linking oligonucleotides. Some monomers need to be converted to amino or hydroxyl that can function as an active site by functionalization reaction. For example, haloalkylstyrene needs to be converted to hydroxyl by hydrolysis or converted to primary amino by Gabriel reaction to become an active site for linking oligonucleotides. Some monomers need to link a linking arm that has amino or hydroxyl as an active site. For example, haloalkylstyrene can react with 4-hydroxybenzoic acid or 4-hydroxyphenylacetic acid to generate hydroxyl, which can become an active site for linking oligonucleotides. Some monomers need to obtain amino or hydroxyl that can be an active site by a combination of the above reactions. For example, 4-vinylphenyl ester monomers need to be hydrolyzed to expose hydroxyl, and then react with hydroquinone or p-phenylenediamine to obtain amino or hydroxyl that can be an active site.

[0020] In some embodiments, the modified monomer is a monomer having a double bond capable of participating in radical polymerization and containing a functional group such as cyano, an ester group, or an amide group, including, but not limited to, acrylonitrile, methacrylonitrile, fumaronitrile, 1,4-dicyano-2-butene, methyl methacrylate, and acrylamide.

[0021] In some embodiments, the initiator is an organic peroxide or azo compound, including but not limited to benzoyl peroxide, lauroyl peroxide, t-butyl peroxyoctoate, 2,2'-azobis(2-methylpropionitrile), 2,2'-azobis(2-methylbutyronitrile), 2,2'-azobis(2,4-dimethylvaleronitrile), etc. The amount of initiator used is 0.5-5% of the total weight of the monomers.

[0022] In some embodiments, the pore-forming agent is an organic solvent or surfactant that does not participate in the polymerization reaction and is insoluble or slightly soluble in water. The pore-forming agent is an aromatic hydrocarbon, such as benzene, toluene, ethylbenzene; an aliphatic hydrocarbon, such as a linear, branched, or cyclic alkane having 6 to 12 carbon atoms, such as hexane, heptane, octane, dodecane, isooctane, isododecane, cyclohexane, etc.; a halohydrocarbon, such as chloroform, chlorobenzene; an ester having 4 or more carbon atoms, such as ethyl acetate, butyl acetate, dibutyl phthalate, etc.; an alcohol, such as a linear, branched, or cyclic alkane alcohol having 4 to 12 carbon atoms, such as hexanol, cyclohexanol, octanol, isooctanol, decanol, dodecanol; an oil-soluble and combinations of one or more of the following surfactants: sorbitan trioleate, polyoxyethylene sorbitol beeswax derivatives, sorbitan tristearate, polyoxyethylene sorbitol hexastearate, ethylene glycol fatty acid esters, propylene glycol fatty acid esters, propylene glycol monostearate, sorbitan sesquioleate, polyoxyethylene sorbitol oleate, glyceryl monostearate, hydroxylanolin, sorbitan monooleate, and propylene glycol monolaurate.

[0023] In some embodiments, the aqueous phase comprises water, a dispersant and an inorganic salt, and the dispersant is a water-soluble polymer, and includes, but is not limited to, one or more of polyvinyl alcohol, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, sodium polyacrylate, and polyvinylpyrrolidone. The amount of the dispersant used is 0.1 to 5% of the aqueous phase weight. The inorganic salt adjusts the density of the aqueous phase and reduces the solubility of each component in the oil phase in the aqueous phase, thereby allowing the oil droplets to be more stably dispersed in the aqueous phase. The inorganic salt includes, but is not limited to, one or more of sodium chloride, potassium chloride, calcium chloride, sodium sulfate, potassium sulfate, calcium sulfate, and the like. The amount of the inorganic salt used is 20% or less of the amount of the aqueous phase used.

[0024] In some embodiments, the weight ratio of the oil phase to the water phase is 1:3 to 1:20 in order to reduce adhesion between resins, promote the conduction of polymerization heat, increase the utilization rate of the equipment, and improve production efficiency.

[0025] In some embodiments of the invention, the components are present in the following amounts: the monovinyl compound initially present in the oil phase comprises 45-95% by weight of the total monomers, the crosslinking monomer initially present in the oil phase comprises 2.9-20% by weight of the total monomers, the functional monomer initially present in the oil phase comprises 2-20% by weight of the total monomers, the modifying monomer initially present in the oil phase comprises 0.1-15% by weight of the total monomers, and the weight of the pore-forming agent initially present in the oil phase is 15-130% of the total monomer weight.

[0026] In some more preferred embodiments of the invention, each component is present in the following amounts: the monovinyl compound initially present in the oil phase comprises 62-86% by weight of the total amount of monomers, the crosslinking monomer initially present in the oil phase comprises 7-13% by weight of the total amount of monomers, the functional monomer initially present in the oil phase comprises 5-15% by weight of the total amount of monomers, the modifying monomer initially present in the oil phase comprises 2-10% by weight of the total amount of monomers, and the weight of the pore-forming agent initially present in the oil phase is 30-110% of the total weight of monomers.

[0027] In some embodiments of the present invention, the polymerization temperature is 50 to 90°C, preferably 70 to 85°C.

[0028] In some embodiments of the present invention, the method for producing the solid phase synthesis support comprises: The method includes the steps of: adding a certain amount of purified water to a reactor, adding and dissolving 0.1-5% by weight of a dispersant and 20% by weight or less of an inorganic salt relative to the weight of the aqueous phase to obtain an aqueous phase; weighing out and uniformly mixing 45-95% by weight of a monovinyl compound, 2.9-20% by weight of a crosslinking monomer, 2-20% by weight of a functional monomer, 0.1-15% by weight of a modified monomer, 15-130% by weight of a pore-forming agent, and an initiator relative to the total amount of monomers so that the weight ratio of the oil phase to the aqueous phase is 1:3 to 1:20, to obtain an oil phase; adding the oil phase to a reactor, stirring, and heating to 50-90°C to cause a reaction; removing the pore-forming agent after the reaction is completed, sieving to collect resins of appropriate particle sizes, vacuum drying to obtain a porous polymer resin, and further reacting the resin to obtain a solid-phase synthesis support having amino or hydroxy functional groups.

[0029] In some embodiments of the present invention, the method for producing the solid phase synthesis support comprises: The method includes the steps of: adding a certain amount of purified water to a reactor, adding and dissolving 0.1-5% by weight of a dispersant and 20% by weight or less of an inorganic salt relative to the weight of the aqueous phase to obtain an aqueous phase; weighing out and uniformly mixing 62-86% by weight of a monovinyl compound, 7-13% by weight of a crosslinking monomer, 5-15% by weight of a functional monomer, 2-10% by weight of a modified monomer, 30-110% by weight of a pore-forming agent, and an initiator so that the weight ratio of the oil phase to the aqueous phase is 1:3-1:20; and adding the oil phase to a reactor, stirring, and heating to 70-85°C to cause a reaction. After the reaction is completed, the pore-forming agent is removed, and the resin of an appropriate particle size is collected by sieving, and vacuum-dried to obtain a porous polymer resin. The resin is further reacted to obtain a solid-phase synthesis support having amino or hydroxy functional groups.

[0030] The above method can provide the solid-phase synthesis support of the present invention, i.e., the porous resin containing hydroxy or amino. In the present invention, the degree of substitution, i.e., the content of hydroxy or amino, can be calculated by reacting with Fmoc-Leu-OH, removing the Fmoc protecting group, and determining the amount of Fmoc removed by colorimetry to calculate the content of amino or hydroxy in the support.

[0031] In some examples, the specific operation is as follows: 1.0 g of the carrier is accurately weighed and suspended in 7 ml of acetonitrile solution, then 0.5 g of Fmoc-Leu-OH, 0.5 g of HBTU, and 0.5 ml of DIEA are added, and the reaction is carried out at room temperature with stirring for 2 h. After the reaction is completed, the resin is washed successively with acetonitrile (10 ml per use, washed twice), and methanol (10 ml per use, washed three times), and then the resin is baked. 0.1000 g of the resin is accurately weighed and suspended in a 20% piperidine / DMF (v / v) solution, shaken at room temperature for 30 min, filtered, and the filtrate is collected, and the resin is washed with DMF and the filtrate is collected. The filtrate is combined, adjusted to a constant volume, and diluted by an appropriate factor, and the absorbance at 300 nm is measured. A series of Fmoc-Leu-OH with known concentrations is used to carry out the same Fmoc removal reaction, and the absorbance is measured to create a calibration curve. The amino or hydroxy content in the carrier is calculated from the calibration curve.

[0032] The functional group content of the carrier is determined by the ratio of the functional monomer in the monomer weight, and a series of carriers with different hydroxyl or amino contents can be obtained by adjusting the amount of the functional monomer used. The functional group content of the carrier determines the synthesis amount of oligonucleotide, and if the functional group content is too low, the yield of one rod of oligonucleotide decreases, and if the functional group content is too high, it affects the purity of the oligonucleotide. In the present invention, the functional group content of the carrier is in the range of 100 to 1000 μmmol / g, preferably 400 to 700 μmmol / g.

[0033] In the present invention, the particle size of the carrier is detected by a particle image processing device, that is, the carrier is uniformly distributed on a slide glass, the carrier particles are magnified by a microscope, the image of the carrier particles magnified by the microscope is taken by a camera, and the morphological characteristics and particle size of the carrier are analyzed and calculated by a computer.

[0034] The size of the carrier particle size mainly depends on the type and amount of dispersant in the aqueous phase, the type and amount of pore-forming agent, and the rotation speed of the stirring in the suspension polymerization process. By adjusting these conditions, the particle size of the carrier can be adjusted. If the carrier particle size is too large, the specific surface area of ​​the carrier decreases, the number of active sites per unit surface area increases, and the purity of the oligonucleotide is affected, while the mass transfer rate during the synthesis of the oligonucleotide slows down and impurities increase. If the carrier particle size is too small, the pressure during the synthesis process is too high, and the cost of the equipment increases. In the present invention, the particle size of the carrier is in the range of 35 to 200 μm, preferably 50 to 100 μm.

[0035] The average pore size of the carrier is measured by mercury intrusion porosimetry. That is, 0.1500 to 0.3000 g of the sample is accurately weighed and placed in a fully automatic mercury intrusion apparatus AutoPoreIV9500 (Micromeritics Instrument Co.), and measured by mercury intrusion porosimetry under conditions of mercury contact angle of 130° and surface tension of 485 dyn / cm. The size of the average pore size of the carrier mainly depends on the type and amount of pore-forming agent, the amount of crosslinking agent, reaction temperature and time, etc. By adjusting these conditions, the average pore size of the carrier can be adjusted. If the average pore size of the carrier is too small, material transfer becomes difficult, affecting the synthesis efficiency, and if the average pore size of the carrier is too large, the specific surface area of ​​the carrier decreases and the active sites per unit area increase, and in the oligonucleotide synthesis process, if the nucleosides increase, they affect each other and affect the purity of the oligonucleotide. In the present invention, the average pore size of the carrier is 10 to 200 nm, preferably 40 to 100 nm. Effect of the Invention

[0036] Compared with the prior art, the present invention has three main advantages. First, the present invention uses a monomer with a reactivity ratio close to that of styrene as a crosslinker, which is favorable for improving the uniformity of the chemical structure inside the resin and forming uniformly distributed active sites. Second, the improvement of the uniformity of the chemical structure inside the resin is favorable for forming uniform channels inside the resin and reducing mass transfer resistance. Third, compared with divinylbenzene, the structure of such a crosslinker has flexible molecular chains, which can improve the swelling of the resin in different organic solvents. [Brief description of the drawings]

[0037] [Figure 1] 1 is a scanning electron microscope image of the carrier in Example 1. [Diagram 2] 1 is a scanning electron microscope image of the carrier in Example 2. [Diagram 3] 1 is a scanning electron microscope image of the carrier in Example 3. [Figure 4]1 is a scanning electron microscope image of the carrier in Example 4. [Diagram 5] 1 is a scanning electron microscope image of the carrier in Example 5. [Figure 6] 1 is a scanning electron microscope image of the carrier in Example 6. [Figure 7] 1 is a scanning electron microscope image of the carrier in Example 7. [Figure 8] 1 is a scanning electron microscope image of the carrier in Example 8. [Figure 9] 1 is a scanning electron microscope image of the carrier in Example 9. [Figure 10] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 1. [Figure 11] 4 shows the pore size distribution of the support obtained by mercury porosimetry in Example 2. [Figure 12] 4 shows the pore size distribution of the support obtained by mercury porosimetry in Example 3. [Figure 13] 4 shows the pore size distribution of the support obtained by mercury porosimetry in Example 4. [Figure 14] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 5. [Figure 15] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 6. [Figure 16] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 7. [Figure 17] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 8. [Figure 18] 1 shows the pore size distribution of the support obtained by mercury porosimetry in Example 9. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] The technical solutions of the present invention are further illustrated below with reference to specific examples, but the present invention is not limited to these examples. EXAMPLES

[0039] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 108.8g of styrene, 14g of 1,2-di(p-vinylphenyl)ethane, 12.2g of p-chloromethylstyrene, 5g of fumaronitrile, 6g of sorbitan trioleate, 40g of isooctanol, 20g of isododecane, and 2.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 80°C, and polymerized for 6h. After the reaction was completed, the mixture was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymeric porous resin with a chlorine content of 565μmmol / g. 50g of polymeric porous resin and 500ml of N,N-dimethylformamide were added to a 1L reactor equipped with a condenser, a stirrer and a thermometer, and stirred. Then, 30g of phthalimide potassium salt was added, and the temperature was raised to 95°C and reacted for 16 hours. After the reaction was completed, it was cooled to room temperature, and then washed twice with N,N-dimethylformamide, washed with purified water until neutral, and then washed three times with absolute ethanol, and the resin was filtered and dried. 200g of absolute ethanol and 50g of hydrazine hydrate were added to the reactor, and the temperature was raised to 75°C and reacted for 16 hours. After the reaction was completed, it was washed three times with a volume ratio of 50:50 ethanol / purified aqueous solution, washed with purified water until neutral, and then washed three times with absolute ethanol, and the washing liquid was filtered. 200g of absolute ethanol and 50g of concentrated hydrochloric acid were added to the reactor, and the temperature was raised to 60°C and reacted for 6 hours. After the reaction was completed, the mixture was cooled to room temperature, washed with water until neutral, and then dried in a vacuum to obtain a solid-phase synthesis support containing amino. The obtained amino solid-phase synthesis support had an amino content of 554 μmmol / g and an average pore diameter of 54 nm measured by mercury intrusion porosimetry. EXAMPLES

[0040] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 77g of methylstyrene, 28g of di(p-vinylphenyl)methane, 14g of p-chloromethylstyrene, 21g of 1,4-dicyano-2-butene, 15g of sorbitan monooleate, 60g of isooctanol, 30g of dibutyl phthalate, and 1g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 70°C, and polymerized for 8h. After the reaction was completed, the mixture was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymeric porous resin with a chlorine content of 646μmmol / g. 50g of polymeric porous resin and 500ml of N,N-dimethylformamide were added to a 1L reactor equipped with a condenser, a stirrer and a thermometer, and stirred. Then, 35g of potassium phthalimide salt was added, and the temperature was raised to 95°C and reacted for 16 hours. After the reaction was completed, it was cooled to room temperature, and then washed twice with N,N-dimethylformamide, washed with purified water until neutral, and then washed three times with absolute ethanol, and the resin was filtered and dried. 200g of absolute ethanol and 50g of hydrazine hydrate were added to the reactor, and the temperature was raised to 75°C and reacted for 16 hours. After the reaction was completed, it was washed three times with a volume ratio of 50:50 ethanol / purified aqueous solution, washed with purified water until neutral, and then washed three times with absolute ethanol, and the washing liquid was filtered. 200g of absolute ethanol and 50g of concentrated hydrochloric acid were added to the reactor, and the temperature was raised to 60°C and reacted for 6 hours. After the reaction was completed, the mixture was cooled to room temperature, washed with water until neutral, and then vacuum dried to obtain an amino-containing solid-phase synthesis support. The amino content of the obtained amino solid-phase synthesis support was 635 μmmol / g, and the average pore diameter measured by mercury intrusion porosimetry was 145 nm. EXAMPLES

[0041] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 123g of ethylstyrene, 8g of 1,3-di(p-vinylphenyl)propane, 8.8g of 4-(4-bromobutyl)styrene, 0.2g of acrylonitrile, 3g of polyoxyethylene sorbitol hexastearate, 18g of isododecane, 5g of dibutyl phthalate, and 2g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 65°C, and polymerized for 10h. After the reaction was completed, the mixture was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymeric porous resin with a bromine content of 257μmmol / g. 50g of polymeric porous resin and 600ml of N,N-dimethylformamide were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and stirred. Then, 5.4g of 4-hydroxybenzoic acid, 5.4g of anhydrous potassium carbonate and 0.3g of potassium iodide were added. The temperature was raised to 75°C, and the reaction was carried out for 6 hours. After the reaction was completed, the mixture was cooled to room temperature, washed with water until neutral, and then dried in vacuum to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 250μmmol / g and an average pore diameter measured by mercury intrusion porosimetry of 23nm. EXAMPLES

[0042] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 112g of styrene, 9g of 1,4-bis(4'-vinylphenoxy)butane, 9g of p-chloromethylstyrene, 10g of acrylamide, 1g of ethylene glycol fatty acid ester, 30g of toluene, 60g of dibutyl phthalate, and 4.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 60°C, and polymerized for 7h. After the reaction was completed, the mixture was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymeric porous resin with a chlorine content of 418μmol / g. 50g of polymeric porous resin and 300ml of anhydrous ethanol were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and stirred. 30g of sodium hydroxide was weighed into a beaker, dissolved in 300ml of deionized water, and slowly added to the reactor. The temperature was raised to 65°C and the reaction was allowed to proceed for 6 hours. After the reaction was completed, the mixture was cooled to room temperature, washed with water until neutral, and then vacuum dried to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 410μmmol / g and an average pore diameter measured by mercury intrusion porosimetry of 38nm. EXAMPLES

[0043] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 83g of styrene, 23g of 1,2-di(p-vinylphenyl)ethane, 18g of N-(4-vinylphenyl)-acetamide, 14g of fumaronitrile, 2g of sorbitan sesquioleate, 10g of dodecanol, 10g of cyclohexane, and 2g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 55°C, and polymerized for 10h. After the reaction was completed, the mixture was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resins with particle sizes of 50 to 100μm, and vacuum-dried to obtain a polymeric porous resin. 50g of polymeric porous resin and 300ml of anhydrous ethanol were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and stirred. 30g of sodium hydroxide was weighed into a beaker, dissolved in 300ml of deionized water, and slowly added to the reactor. The temperature was raised to 65°C and reacted for 6 hours. After the reaction was completed, the mixture was cooled to room temperature, washed with water until neutral, and then dried in vacuum to obtain a solid-phase synthesis support containing amino. The obtained amino solid-phase synthesis support had an amino content of 822μmmol / g and an average pore diameter measured by mercury intrusion porosimetry of 34nm. EXAMPLES

[0044] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 107.7g of styrene, 12.8g of 1,2-di(p-vinylphenyl)ethane, 12.5g of 4-acetoxystyrene, 7g of fumaronitrile, 10g of sorbitan trioleate, 42g of isooctanol, 21g of isododecane, and 2.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 78°C, and polymerized for 6h. After the reaction was completed, the resin was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymer porous resin. 50g of polymeric porous resin and 300ml of acetonitrile were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and the mixture was stirred. 7.5ml of hydrazine hydrate was slowly added, and the mixture was reacted at room temperature for 3h. After the reaction was completed, the mixture was washed with water until neutral, and then vacuum dried to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 538μmmol / g, and an average pore diameter of 58nm measured by mercury intrusion porosimetry. EXAMPLES

[0045] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 128g of styrene, 8g of 1,3-di(p-vinylphenyl)propane, 3g of 4-acetoxystyrene, 1g of fumaronitrile, 0.4g of polyoxyethylene sorbitol hexastearate, 35g of dibutyl phthalate, 12g of toluene, and 3.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 70°C, and polymerized for 6h. After the reaction was completed, the resin was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100 μm, and vacuum dried to obtain a polymer porous resin. 50g of polymeric porous resin and 300ml of acetonitrile were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and the mixture was stirred. 7.5ml of hydrazine hydrate was slowly added, and the mixture was allowed to react at room temperature for 3h. After the reaction was completed, the mixture was washed with water until neutral, and then vacuum dried to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 125μmmol / g, and an average pore diameter measured by mercury intrusion porosimetry of 46nm. EXAMPLES

[0046] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 83g of styrene, 17g of 1,4-bis(4'-vinylphenoxy)butane, 27g of benzoyloxystyrene, 13g of fumaronitrile, 103g of isooctanol, 51g of isododecane, and 3g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 80°C, and polymerized for 6h. After the reaction was completed, the resin was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymer porous resin. 50g of polymeric porous resin and 300ml of acetonitrile were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and the mixture was stirred. 7.5ml of hydrazine hydrate was slowly added, and the mixture was reacted at room temperature for 3h. After the reaction was completed, the mixture was washed with water until neutral, and then vacuum dried to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 843μmmol / g, and an average pore diameter measured by mercury intrusion porosimetry of 42nm. EXAMPLES

[0047] 2L of purified water, 20g of polyvinyl alcohol, and 30g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 85g of styrene, 24g of 1,2-di(p-vinylphenyl)ethane, 18g of methyl 4-vinylbenzoate, 13g of fumaronitrile, 4g of propylene glycol monolaurate, 40g of dibutyl phthalate, 40g of isododecane, and 2.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 70°C, and polymerized for 6h. After the reaction was completed, the resin was washed with hot water, extracted by refluxing with ethanol, sieved to collect resin with a particle size of 50 to 100 μm, and vacuum dried to obtain a polymeric porous resin. 50g of polymeric porous resin and 300ml of acetonitrile were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and the mixture was stirred. Then, 8.8g of hydroquinone, 23g of HBTU and 13ml of DIEA were added, and the mixture was reacted at room temperature for 2h. After the reaction was completed, the mixture was washed with water until neutral, and then dried in vacuum to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 724μmmol / g and an average pore diameter measured by mercury intrusion porosimetry of 124nm.

[0048] [Comparative Example 1] 2L of purified water, 20g of polyvinyl alcohol, and 60g of sodium chloride were added to a 3L reactor equipped with a condenser, a stirrer, and a thermometer, and dissolved to obtain an aqueous phase. 111g of styrene, 11g of divinylbenzene (content 80% by weight), 13g of 4-acetoxystyrene, 5g of fumaronitrile, 8g of sorbitan monooleate, 40g of isooctanol, 20g of isododecane, and 2.5g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase was added to the reactor, stirred, heated to 78°C, and polymerized for 6h. After the reaction was completed, the resin was washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect resin with a particle size of 50 to 100μm, and vacuum dried to obtain a polymer porous resin. 50g of polymeric porous resin and 300ml of acetonitrile were added to a 1L reactor equipped with a condenser, stirrer and thermometer, and the mixture was stirred. 7.5ml of hydrazine hydrate was slowly added, and the mixture was reacted at room temperature for 3h. After the reaction was completed, the mixture was washed with water until neutral, and then vacuum dried to obtain a solid-phase synthesis support containing hydroxyl. The obtained hydroxyl solid-phase synthesis support had a hydroxyl content of 550μmmol / g, and an average pore diameter measured by mercury intrusion porosimetry of 64nm. EXAMPLES

[0049] The swelling degree of each of the solid phase synthesis supports produced in Examples 1 to 9 in acetonitrile and toluene was tested. The test was performed as follows. Approximately 1.5 g of a sample was weighed out and placed in a stoppered graduated cylinder, and toluene or acetonitrile was added to the desired interval, and then the resin and solvent were stirred using a glass rod to allow sufficient swelling, the stopper was tightened, and after 2 to 3 hours, the resin was gently stirred using a glass rod to remove air bubbles and to uniformly disperse the resin to prevent caking, the stirring rod was removed, and the graduated cylinder was struck on a table equipped with a rubber mat to pile the resin tightly, and after leaving it for 24 hours, the volume was removed and the swelling degree was calculated. The results are shown in Table 1. [Table 1] EXAMPLES

[0050] Oligonucleotide fragments were synthesized using the solid-phase synthesis supports prepared in Examples 1, 2, 6, and 9 and Comparative Example 1, and NittoPhase HL solid-phase synthesis support, and the performance of the resin was evaluated. In order to more clearly demonstrate the superiority of the present invention, the carrier loading amount (g) = synthesis column volume (ml) / carrier swelling degree in toluene (ml / g), and in the oligonucleotide synthesis process, the washing volume was limited to one synthesis column volume. 10 g of solid-phase synthesis support and 50 ml of acetonitrile were weighed into a reactor and swollen for 10 min, after which 3.0 g of DMT-dT-3'-succinic acid, 1.5 g of HBTU, and 1.3 ml of DIEA were added and reacted at room temperature for 12 h. After the reaction was completed, the support was washed five times with acetonitrile, and then Cap A (20 ml of acetonitrile, 7.5 ml of pyridine, and 5.0 ml of N-methylimidazole) and Cap B (10 ml of acetonitrile, 4 ml of acetic anhydride) were added and reacted at room temperature for 30 min. After the reaction was completed, the support was washed five times with acetonitrile and dried in vacuum to obtain a support carrying DMT-dT. The supported DMT group was removed using a paratoluenesulfonic acid / acetonitrile solution, and the amount of DMT group supported on the support was measured by spectrophotometry at a wavelength of 412 nm, and the results are shown in Table 2. The carrier carrying DMT-dT was weighed and packed into a synthesis column (32 ml), and the synthesis column was attached to an AKTA OligoPilot 100 to synthesize a 20-base-long oligonucleotide with the sequence d[ACGTACGTACGTACGTACGT]. The synthesis was carried out as follows: 1. The resin was swollen with dichloromethane. 2. The DMT group was removed with 10% DCA / DCM. 3. The resin was washed with anhydrous acetonitrile. 4. A phosphoramidite monomer and an activating reagent were added and condensed. 5. The resin was washed with anhydrous acetonitrile. 6. An oxidizing agent was added and oxidation was performed. 7. The resin was washed with anhydrous acetonitrile. 8. A capping reagent was added and capping was performed. 9. The resin was washed with anhydrous acetonitrile. 10. Step 2 was repeated to start the next cycle. After the synthesis was completed, the support was removed and dried. It was then placed in a glass bottle, and an appropriate amount of concentrated aqueous ammonia was added and reacted at 55°C for 16 hours to decompose and remove the oligonucleotide from the support, and to remove the protecting groups on the bases. The support and oligonucleotide were separated by filtration, and the filtrate was dried to obtain crude oligonucleotide powder, whose purity was detected by HPLC, and the oligonucleotide yield was calculated. The results are shown in Table 2. [Table 2]

[0051] As can be seen from Table 2, the use of the oligonucleotide solid-phase synthesis support of the present invention can increase the yield and purity of oligonucleotide, which is advantageous for reducing the production cost of oligonucleotide.

[0052] It should be noted that the above is merely a preferred embodiment of the present invention, and a person skilled in the art may make multiple modifications and improvements without departing from the concept of the present invention, all of which are intended to be included in the patent scope of the present invention.

Claims

1. A solid phase synthesis support comprising: The solid-phase synthesis support is a copolymer having a skeleton having repeating structural units represented by formula (I), formula (II), formula (III) and formula (IV), 【Chemistry 1】 Here, R ３ -H, -CN, and -CH ２ -CN; R ４ is -H or -CH ３ and R ５ -CN, -CH ２ -CN, -COOCH ３ and -CONH ２ is selected from the group consisting of 【Chemistry 2】 R ６ Ha-(CH ２ ) x-, where x is 2; 【Chemistry 3】 R ７ H, CH ３ (CH ２ ) z- or CH ３ (CH ２ )zO-(z is an integer of 0 to 4), (CH ３ ) ２ CH-, (CH ３ ) ２ CH (CH ２ ) -, (CH ３ ) ２ CH (CH ２ ) ２ -, (CH ３ ) ３ C-, CH ３ CH ２ CH (CH ３ ) -, CH ３ CH ２ C (CH ３ ) ２ - and CH ３ CH ２ CH ２ CH (CH ３ )- selected from the group consisting of 【Chemistry 4】 ( ８ は!HHHH ２ !!、!H ２ 、!| ２ . ２ 、!| ２ _____________ ６ _ ４ H、!H ２ _____________ ２  ６ _ ４ H、((H ２ ) ４ _____________ ６ _ ４ H、((H ２ ) ４ _____________ ２  ６ _ ４ ___、______________________ ６ _ ４ _______________________ H ２ COO-C ６ H ４ -OH; wherein the solid phase synthesis support has a hydroxy or amino content of 100-1000 μmol / g, and the amino or hydroxy content is obtained by reacting with Fmoc-Leu-OH, followed by removing the Fmoc protecting group, determining the amount of the removed Fmoc group by colorimetry, and then calculating the amino or hydroxy content in the support; The solid-phase synthesis support has a particle size of 35 to 200 μm when detected by a particle image processing device, that is, when the support is uniformly distributed on a slide glass and observed under a microscope while an image of the enlarged particles of the support is taken by a camera, and the morphological characteristics and particle size of the support are analyzed and calculated by a computer. The solid phase synthesis support has an average pore diameter of 10 to 200 nm, as measured by mercury intrusion porosimetry under the following conditions: 0.1500 to 0.3000 g of a sample is accurately weighed and placed in a fully automatic mercury intrusion apparatus AutoPoreIV9500 (manufactured by Micromeritics) with a mercury contact angle of 130° and a surface tension of 485 dyn / cm. A solid phase synthesis support characterized by:

2. 2. The solid phase synthesis support according to claim 1, characterized in that the content of hydroxy or amino is in the range of 400-700 μmol / g, said content of amino or hydroxy is obtained by reacting with Fmoc-Leu-OH, followed by removing the Fmoc protecting group, determining the amount of removed Fmoc group by colorimetric method, and then calculating said content of amino or hydroxy in the support.

3. The solid-phase synthesis support according to claim 1, characterized in that the solid-phase synthesis support is detected by a particle image processing device, i.e., the support is uniformly distributed on a slide glass and observed under a microscope while taking an image of the enlarged particles of the support with a camera, and the morphological characteristics and particle size of the support are analyzed and calculated by a computer, and the particle size range is 50 to 100 μm.

4. The solid-phase synthesis support according to claim 1, characterized in that the average pore size of the solid-phase synthesis support is 40 to 100 nm when measured by mercury intrusion porosimetry under the following conditions: 0.1500 to 0.3000 g of a sample is accurately weighed, placed in a fully automatic mercury intrusion apparatus AutoPoreIV9500 (manufactured by Micromeritics), and the contact angle of mercury is 130° and the surface tension is 485 dyn / cm.

5. A method for producing a solid phase synthesis support comprising steps A and B, Step A: a step of preparing an aqueous phase consisting of water, a dispersant and an inorganic salt, and an oil phase consisting of a cross-linking monomer, a monovinyl compound, a functional monomer, a modified monomer, a pore-forming agent and an initiator, respectively, wherein the cross-linking monomer, the monovinyl compound, the functional monomer and the modified monomer are monomers capable of polymerization reaction; wherein the cross-linking monomer is 1,2-di(4-vinylphenyl)ethane; The monovinyl compound is a styrene having an unsubstituted or alkyl or alkoxy substitution on the benzene ring having 1 to 5 carbon atoms; the functional monomer is selected from the group consisting of 4-hydroxystyrene, 4-hydroxymethylstyrene, 4-acetoxystyrene, benzoyloxystyrene, 4-aminostyrene, 4-aminomethylstyrene, 4-(4-bromobutyl)styrene, p-chloromethylstyrene, methyl 4-vinylbenzoate, and ethyl 4-vinylbenzoate; the modifying monomer is selected from the group consisting of acrylonitrile, methacrylonitrile, fumaronitrile, 1,4-dicyano-2-butene, methyl methacrylate, and acrylamide; The pore-forming agent is one or a combination of a plurality of pore-forming agents selected from the group consisting of benzene, toluene, ethylbenzene, hexane, heptane, octane, dodecane, isooctane, isododecane, cyclohexane, chloroform, chlorobenzene, ethyl acetate, butyl acetate, dibutyl phthalate, hexanol, cyclohexanol, octanol, isooctanol, decanol, dodecanol, sorbitan trioleate, polyoxyethylene sorbitol beeswax, sorbitan tristearate, polyoxyethylene sorbitol hexastearate, ethylene glycol fatty acid esters, propylene glycol fatty acid esters, propylene glycol monostearate, sorbitan sesquioleate, polyoxyethylene sorbitol oleate, glyceryl monostearate, hydroxylanolin, sorbitan monooleate, and propylene glycol monolaurate; The dispersant is a water-soluble polymer selected from the group consisting of polyvinyl alcohol, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, carboxymethyl cellulose, sodium polyacrylate, and polyvinylpyrrolidone; Step B: a step of adding the oil phase to the aqueous phase, stirring and heating the mixture to cause a polymerization reaction, and removing the pore-forming agent after the polymerization reaction is completed to obtain a porous polymer resin; wherein the porous polymeric resin is further reacted to obtain a solid phase synthesis support containing hydroxy or amino functional groups; The solid phase synthesis support has a hydroxy or amino content of 100 to 1000 μmol / g; The solid-phase synthesis support is detected by a particle image processing device, that is, the solid-phase synthesis support is uniformly distributed on a slide glass and observed under a microscope while taking an image of the enlarged particles of the solid-phase synthesis support with a camera, and the morphological characteristics and particle size of the solid-phase synthesis support are analyzed and calculated by a computer, and the solid-phase synthesis support has a particle size range of 35 to 200 μm, The solid phase synthesis support has an average pore size of 10 to 200 nm; A method for producing a solid phase synthesis support comprising the steps of:

6. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the initiator is an organic peroxide or an azo compound.

7. The method for producing a solid phase synthesis support according to claim 6, characterized in that the initiator is selected from the group consisting of benzoyl peroxide, lauroyl peroxide, t-butyl peroxyoctoate, 2,2'-azobis(2-methylpropionitrile), 2,2'-azobis(2-methylbutyronitrile), and 2,2'-azobis(2,4-dimethyl)valeronitrile.

8. In the aqueous phase, the dispersant is 0.1 to 5% by weight, and the inorganic salt is 20% by weight or less. The weight ratio of the oil phase to the water phase is 1:3 to 1:20; In the oil phase, the monovinyl compound accounts for 45 to 95% by weight of the total amount of monomers, In the oil phase, the cross-linking monomer accounts for 2.9 to 20% by weight of the total amount of monomers; In the oil phase, the functional monomer accounts for 2 to 20% by weight of the total amount of monomers; In the oil phase, the modifying monomer accounts for 0.1 to 15% by weight of the total amount of monomers; In the oil phase, the weight of the pore-forming agent is 15 to 130% of the total weight of the monomers. The method for producing a solid phase synthesis support according to claim 5 .

9. The method for producing a solid phase synthesis support according to claim 5, wherein the polymerization reaction is carried out at 50 to 90°C.

10. The manufacturing method includes: (i) adding a certain amount of purified water to a reactor, and adding and dissolving 0.1 to 5% by weight of a dispersant and 20% by weight or less of an inorganic salt based on the weight of the aqueous phase to obtain an aqueous phase; a step of weighing out and uniformly mixing 45 to 95% by weight of a monovinyl compound, 2.9 to 20% by weight of a crosslinking monomer, 2 to 20% by weight of a functional monomer, 0.1 to 15% by weight of a modified monomer, 15 to 130% by weight of a pore-forming agent, and an initiator, based on the total amount of monomers, so that the weight ratio of the oil phase to the aqueous phase is 1:3 to 1:20, to obtain an oil phase; Adding the oil phase to a reactor, stirring and heating to 50-90°C to carry out a reaction, removing the pore-forming agent after the reaction is completed, collecting resin with a suitable particle size by sieving, and drying under vacuum to obtain a porous polymer resin, and further reacting the resin to obtain a solid phase synthesis support with amino or hydroxy functional groups; or (ii) adding a certain amount of purified water to a reactor, and adding and dissolving 0.1 to 5% by weight of a dispersant and 20% by weight or less of an inorganic salt based on the weight of the aqueous phase to obtain an aqueous phase; a step of weighing out and uniformly mixing 62 to 86% by weight of a monovinyl compound, 7 to 13% by weight of a crosslinking monomer, 5 to 15% by weight of a functional monomer, 2 to 10% by weight of a modified monomer, 30 to 110% by weight of a pore-forming agent, and an initiator, based on the total amount of monomers, so that the weight ratio of the oil phase to the aqueous phase is 1:3 to 1:20; Add the oil phase to a reactor, stir, and heat to 70-85°C to react; after the reaction is completed, remove the pore-forming agent, and collect the resin with the appropriate particle size by sieving; and dry in vacuum to obtain a porous polymer resin; and further react the resin to obtain a solid-phase synthesis support with amino or hydroxy functional groups; The method for producing a solid phase synthesis support according to any one of claims 5 to 9, comprising:

11. In a 3 L reactor equipped with a condenser, a stirrer, and a thermometer, 2 L of purified water, 20 g of polyvinyl alcohol, and 30 g of sodium chloride were added and dissolved to obtain an aqueous phase. 5 g of fumaronitrile, 6 g of sorbitan trioleate, 40 g of isooctanol, 20 g of isododecane, and 2.5 g of benzoyl peroxide were weighed out and mixed uniformly to obtain an oil phase. Add the oil phase to a reactor, stir, and heat to 80°C for polymerization for 6 hours. After the reaction is completed, wash with hot water, and reflux with ethanol to extract and remove the pore-forming agent. Sieve to collect the resin with a particle size of 50-100 μm, and vacuum dry to obtain a polymeric porous resin with a chlorine content of 565 μmol / g. In a 1 L reactor equipped with a condenser, a stirrer and a thermometer, 50 g of polymeric porous resin and 500 ml of N,N-dimethylformamide were added and stirred, then 30 g of phthalimide potassium salt was added, the temperature was raised to 95° C. and reacted for 16 hours, after the reaction was completed, the mixture was cooled to room temperature, then washed twice with N,N-dimethylformamide, washed until neutral with purified water, and then washed three times with absolute ethanol, the resin was filtered and dried, 200 g of absolute ethanol and 50 g of hydrazine hydrate were added to the reactor, the temperature was raised to 75° C. and reacted for 16 hours, after the reaction was completed, the mixture was cooled to room temperature, and then washed twice with N,N-dimethylformamide, washed until neutral with purified water, and then washed three times with absolute ethanol, the resin was filtered and dried, 200 g of absolute ethanol and 50 g of hydrazine hydrate were added to the reactor, and the temperature was raised to 75° C. and reacted for 16 hours, after the reaction was completed, the mixture was cooled to room temperature, and then washed twice with N,N-dimethylformamide, washed until neutral with purified water, and then washed three times with absolute ethanol, The method for producing the solid-phase synthesis support according to any one of claims 5 to 10, further comprising the steps of: washing three times with a 50:50 volume ratio of ethanol / purified water solution, washing with purified water until neutral, washing three times with absolute ethanol, filtering the washing liquid, adding 200 g of absolute ethanol and 50 g of concentrated hydrochloric acid to a reactor, heating to 60°C and reacting for 6 hours, cooling to room temperature after the reaction is completed, washing with water until neutral, and then drying in a vacuum to obtain a solid-phase synthesis support having an amino content of 554 μmol / g and an average pore diameter of 54 nm measured by mercury intrusion porosimetry.

12. In a 3 L reactor equipped with a condenser, a stirrer, and a thermometer, 2 L of purified water, 20 g of polyvinyl alcohol, and 30 g of sodium chloride were added and dissolved to obtain an aqueous phase. Styrene 107.7 g, 1,2-di(4-vinylphenyl)ethane 12.8 g, 4-a 12.5 g of acetoxystyrene, 7 g of fumaronitrile, 10 g of sorbitan trioleate, 42 g of isooctanol, 21 g of isododecane, and 2.5 g of benzoyl peroxide were weighed and mixed uniformly to obtain an oil phase. The oil phase is added to a reactor, stirred, heated to 78°C and polymerized for 6h. After the reaction is completed, the resin is washed with hot water, refluxed with ethanol to extract and remove the pore-forming agent, sieved to collect the resin with a particle size of 50-100 μm, and vacuum dried to obtain a polymeric porous resin. Add 50 g of the polymeric porous resin to a 1 L reactor equipped with a condenser, a stirrer and a thermometer, add 300 ml of acetonitrile, stir, slowly add 7.5 ml of hydrazine hydrate, react at room temperature for 3 hours, wash with water until neutral after the reaction is completed, and then dry in vacuum to obtain a hydroxy-containing solid-phase synthesis support having a hydroxy content of 538 μmol / g and an average pore diameter of 58 nm measured by mercury intrusion porosimetry; The method for producing a solid phase synthesis support according to any one of claims 5 to 10, comprising:

13. 13. Use of a solid-phase synthesis support according to claim 1, produced by the method according to any one of claims 5 to 12, for solid-phase synthesis of oligonucleotides.

14. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the monovinyl compound accounts for 62 to 86% by weight of the total amount of monomers in the oil phase.

15. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the cross-linking monomer accounts for 7 to 13% by weight of the total amount of monomers in the oil phase.

16. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the functional monomer accounts for 5 to 15% by weight of the total amount of monomers in the oil phase.

17. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the modified monomer accounts for 2 to 10% by weight of the total amount of monomers in the oil phase.

18. 6. The method for producing a solid phase synthesis support according to claim 5, wherein the pore-forming agent accounts for 30 to 110% by weight of the total amount of monomers in the oil phase.

19. The method for producing a solid phase synthesis support according to claim 5, wherein the polymerization reaction is carried out at 70 to 85°C.

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