Method for producing target molecule

Abstract

Provided is a method for producing a target molecule, with which there is little breakage of particles during synthesis of the target molecule and which can enhance reaction efficiency.　The method for producing a target molecule according to the present invention comprises a step for synthesizing a target molecule on a silicone particle.

Description

Methods for producing target molecules

[0001] The present invention relates to a method for producing a molecule of interest.

[0002] A method for synthesizing target molecules such as peptides and nucleic acids by solid-phase synthesis is known. In solid-phase synthesis, target molecules are synthesized on particles (supports). Generally, non-porous, highly swellable low-crosslinked polyethylene particles are widely used as the particles. Non-swellable inorganic porous particles or porous resin particles may also be used as the particles.

[0003] For example, Patent Document 1 below discloses an inorganic porous support having specific silyl groups and a pore size of 20 nm or more as particles that can be used as a support for solid-phase synthesis of nucleic acids.

[0004] Furthermore, Patent Document 2 listed below discloses porous resin beads that are copolymers consisting of monovinyl monomer units and crosslinkable vinyl monomer units and have groups that can bond with carboxy groups through a dehydration condensation reaction, as particles that can be used as supports for solid-phase synthesis of nucleic acids.

[0005] WO2021 / 230293A1 JP 2015-129268 A

[0006] Conventional low-crosslinked polyethylene particles are non-porous and highly swellable, so it takes time for synthesis reagents to diffuse into the interior of the particles, making it difficult to increase the reaction efficiency when synthesizing target molecules.

[0007] On the other hand, with non-swelling particles such as those described in Patent Document 1, it is difficult to increase the reaction efficiency when synthesizing a target molecule because it is not possible to sufficiently increase the number of synthesis initiation points.

[0008] Conventional porous resin particles such as those described in Patent Document 2 can improve the reaction efficiency of target molecule synthesis to a certain extent. However, conventional porous resin particles are prone to cracking during target molecule synthesis. For example, conventional porous resin particles may crack during stirring. If particles crack during target molecule synthesis, it is difficult to separate the cracked particles (particle fragments) from the unbroken particles, and the particle fragments may remain as impurities. As a result, the purity of the target molecule decreases.

[0009] An object of the present invention is to provide a method for producing a target molecule that is less likely to break particles during synthesis of the target molecule and that can increase reaction efficiency.

[0010] Disclosed herein are methods for producing the following molecules of interest.

[0011] Item 1. A method for producing a target molecule, comprising a step of synthesizing the target molecule on silicone particles.

[0012] Item 2. The method for producing a target molecule according to Item 1, wherein the silicone particles are porous silicone particles.

[0013] Item 3. The method for producing a target molecule according to Item 2, wherein the porous silicone particles have an average pore size of 2 nm or more and 300 nm or less.

[0014] Item 4. The compressive modulus of the silicone particles when compressed by 10% is 500 N / mm ２ Item 4. A method for producing a molecule of interest according to any one of Items 1 to 3, wherein the method is:

[0015] Item 5. The method for producing a target molecule according to any one of Items 1 to 4, wherein when the silicone particles are immersed in toluene at 25°C for 24 hours, the swelling ratio represented by the following formula (L) is 1.1 or more and 2.5 or less:

[0016] Swelling ratio = L2 / L1...(L)

[0017] L1: Average particle size of silicone particles before immersion in toluene L2: Average particle size of silicone particles after immersion in toluene for 24 hours

[0018] Item 6. The method for producing a target molecule according to any one of Items 1 to 5, wherein a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group is present on the surface of the silicone particle.

[0019] Item 7. The method for producing a target molecule according to any one of Items 1 to 6, wherein the silicone particles contain a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety.

[0020] Item 8. The method for producing a target molecule according to Item 7, wherein the silicone polymer has a skeleton derived from a (meth)acryloyl group-containing silicone compound (A).

[0021] Item 9. The method for producing a target molecule according to Item 8, wherein the silicone polymer further has a skeleton derived from a radically polymerizable compound (B) different from the (meth)acryloyl group-containing silicone compound.

[0022] Item 10. The method for producing a target molecule according to Item 9, wherein the radical polymerizable compound (B) includes a (meth)acryloyl group-containing compound (B1).

[0023] Item 11. The method for producing a target molecule according to Item 9 or 10, wherein the radical polymerizable compound (B) includes a radical polymerizable compound having a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group.

[0024] Item 12. The method for producing a target molecule according to any one of Items 1 to 11, wherein the silicone particles have an average particle size of 10 μm or more and 500 μm or less.

[0025] Item 13. The specific surface area of ​​the silicone particles is 1 m ２ / g or more 200m ２ Item 13. The method for producing a target molecule according to any one of Items 1 to 12, wherein the total amount of the target molecule is 1000 kJ / g or less.

[0026] Item 14. The method for producing a molecule of interest according to any one of Items 1 to 13, wherein the molecule of interest is a peptide or a nucleic acid.

[0027] The method for producing a target molecule according to the present invention includes a step of synthesizing a target molecule on silicone particles. Because the method for producing a target molecule according to the present invention includes the above-described configuration, the particles are less likely to break during synthesis of the target molecule, and the reaction efficiency can be increased.

[0028] FIG. 1 is a cross-sectional view schematically illustrating an example of a silicone particle used in the present invention.

[0029] The present invention will be described in detail below.

[0030] The method for producing a target molecule according to the present invention comprises a step of synthesizing the target molecule on a silicone particle.

[0031] The method for producing a target molecule according to the present invention has the above-described configuration, so that the particles (silicone particles) are less likely to break during synthesis of the target molecule, and the reaction efficiency can be increased.

[0032] First, the silicone particles that can be used in the present invention will be described.

[0033] (Silicone Particles) The silicone particles contain a silicone polymer. The silicone polymer has a polysiloxane moiety. The silicone particles have appropriate flexibility and strength because of the polysiloxane moiety. Therefore, even if the silicone particles are subjected to an impact, the silicone particles are not likely to break.

[0034] As described below, the silicone polymer preferably has a (meth)acrylic resin moiety. That is, the silicone particles preferably contain a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety. In this case, for example, the silicone particles tend to swell moderately in nonpolar solvents such as toluene or aprotic polar solvents such as dimethylformamide (DMF). Therefore, compared to particles that do not swell, a larger amount of substance can be retained inside. Furthermore, if the amount of substance that can be retained inside can be increased, the number of reaction sites during synthesis of target molecules can be increased, further improving reaction efficiency. Furthermore, when the silicone particles contain a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety, the silicone particles tend to shrink moderately in protic polar solvents such as methanol. Therefore, by changing the solvent, the substance retained inside can be efficiently discharged.

[0035] The silicone particles may be solid silicone particles or porous silicone particles.

[0036] The silicone particles are preferably porous silicone particles. In this case, the amount of substance that can be retained inside can be increased. As a result, the reaction rate during synthesis of the target molecule is increased, and the reaction efficiency can be further improved. Furthermore, while conventional porous resin particles are prone to cracking when subjected to impact, particularly during stirring, the porous silicone particles are less likely to crack even when subjected to impact. Therefore, even when stirring is performed before or after retaining a substance inside, cracking during stirring can be effectively suppressed.

[0037] The present invention will be specifically described below with reference to the drawings.

[0038] FIG. 1 is a cross-sectional view schematically illustrating an example of a silicone particle used in the present invention.

[0039] The silicone particles 1 have a porous structure. The silicone particles 1 are porous particles (porous silicone particles). The silicone particles 1 contain a silicone polymer having a polysiloxane portion and a (meth)acrylic resin portion.

[0040] The compressive modulus of elasticity when the silicone particles are compressed by 10% (10% K value) is preferably 5 N / mm ２ More preferably, 10 N / mm ２ More preferably, 20 N / mm ２ More than 500N / mm ２ or less, more preferably 300 N / mm ２ More preferably, 200 N / mm ２ When the compressive elastic modulus (10% K value) is equal to or greater than the lower limit and equal to or less than the upper limit, the flexibility of the silicone particles can be increased while maintaining strength that will not break, and cracking of the silicone particles can be effectively suppressed.

[0041] The compressive elastic modulus (10% K value) of the silicone particles can be measured as follows.

[0042] Using a micro-compression tester, one silicone particle is compressed with the end face of a smooth cylindrical indenter (diameter 100 μm, made of diamond) under conditions of 25°C, a compression speed of 0.3 mN / sec, and a maximum test load of 20 mN. The load value (N) and compression displacement (mm) at this time are measured. From the obtained measured values, the compressive modulus (10% K value) can be calculated using the following formula. Examples of the micro-compression tester that can be used include the "Micro-Compression Tester MCT-W200" manufactured by Shimadzu Corporation, the "Fisherscope H-100" manufactured by Fischer, and the "ENT-5" manufactured by Elionix. The compressive modulus (10% K value) of the silicone particle is preferably calculated by arithmetically averaging the compressive moduli (10% K value) of 50 arbitrarily selected silicone particles.

[0043] 10% K value (N / mm ２ ) = (3 / 2 １／２ ) F.S. －３／２ ・R －１／２F: Load value (N) when the silicone particle is compressed and deformed by 10%; S: Compression displacement (mm) when the silicone particle is compressed and deformed by 10%; R: Radius of the silicone particle (mm).

[0044] Methods for adjusting the compressive modulus within the preferred range include using porous silicone particles as the silicone particles, adjusting the porosity (specific surface area) of the porous silicone particles, and adjusting the content ratio of the polysiloxane moiety and the (meth)acrylic resin moiety in the silicone polymer.

[0045] When the silicone particles are porous silicone particles, the average pore diameter of the porous silicone particles is preferably 2 nm or more, more preferably 5 nm or more, even more preferably 10 nm or more, preferably 300 nm or less, more preferably 100 nm or less, even more preferably 50 nm or less.When the average pore diameter is above the lower limit and below the upper limit, the flexibility of the porous silicone particles can be increased, so that the cracking of the porous silicone particles can be effectively suppressed.In addition, when the average pore diameter is above the lower limit and below the upper limit, the amount of substance that can be retained inside can be further increased, so that the reaction efficiency when synthesizing target molecules can be further improved.

[0046] The average pore diameter of the porous silicone particles is measured by the BJH method. Nitrogen gas is used as the adsorption gas in the BJH method, and a gas adsorption pore size distribution analyzer (for example, the Nova Touch 4LX manufactured by Quantachrome) is used. An adsorption / desorption isotherm is created by measuring the amount of nitrogen gas adsorbed and desorbed from a measurement sample at relative pressures of 0 to 1, and then the average pore diameter is determined using the BJH method for measurement data at relative pressures of 0.35 or higher during the desorption process.

[0047] The specific surface area of ​​the silicone particles is preferably 1 m ２ / g or more, more preferably 10m ２ / g or more, more preferably 30m ２ / g or more, preferably 200m ２ / g or less, more preferably 150m ２ / g or less, more preferably 125m ２ / g or less, particularly preferably 100m ２ / g or less. When the specific surface area is equal to or greater than the lower limit and equal to or less than the upper limit, the flexibility of the silicone particles can be increased, thereby effectively preventing the silicone particles from cracking. Furthermore, when the specific surface area is equal to or greater than the lower limit and equal to or less than the upper limit, the amount of substance that can be retained inside can be further increased, thereby further improving the reaction efficiency when synthesizing the target molecule.

[0048] The specific surface area of ​​the silicone particles is measured by the multipoint BET method. Nitrogen gas is used as the adsorption gas in the multipoint BET method, and a gas adsorption pore size distribution analyzer (for example, the Nova Touch 4LX manufactured by Quantachrome) is used. An adsorption isotherm is created by measuring the amount of nitrogen gas adsorbed to the measurement sample over a relative pressure range of 0 to 1, and then the specific surface area is determined using the multipoint BET method for the measurement data over a relative pressure range of 0.05 to 0.30 during the adsorption process.

[0049] Methods for adjusting the average pore diameter and the specific surface area to fall within the preferred ranges include using porous silicone particles as the silicone particles and adjusting the amount of porogen used during the production of the porous silicone particles.

[0050] The average particle size of the silicone particles is preferably 10 μm or more, more preferably 25 μm or more, even more preferably 50 μm or more, preferably 500 μm or less, more preferably 250 μm or less, even more preferably 150 μm or less.When the average particle size is above the lower limit and below the upper limit, the flexibility of the silicone particles can be increased, so that the cracking of the silicone particles can be effectively prevented.In addition, when the average particle size is above the lower limit and below the upper limit, the amount of material that can be retained inside can be further increased, so that the reaction efficiency when synthesizing target molecules can be further improved.

[0051] The average particle size of the silicone particles can be determined by observing 100 random silicone particles with a scanning electron microscope (e.g., "Regulus-8220" manufactured by Hitachi High-Technologies Corporation), determining the particle size of each silicone particle using imaging software (e.g., "ImageJ" distributed by the National Institute of Health), and calculating the average particle size. In observation with a scanning electron microscope, the particle size of a single silicone particle is determined as the diameter of the particle if the particle is spherical, and as the longest diameter of the particle if the particle is not spherical.

[0052] The coefficient of variation (CV value) of the particle size of the silicone particles is preferably 25% or less, more preferably 20% or less, and even more preferably 15% or less. When the CV value of the particle size is equal to or less than the upper limit, the variation in the amount of substance that can be retained inside can be reduced. The lower limit of the CV value of the particle size of the silicone particles is not particularly limited. The CV value of the particle size of the silicone particles may be 0% or more, may exceed 0%, may be 1% or more, or may be 3% or more. The range of the CV value of the particle size of the silicone particles can be set by appropriately selecting the lower limit and the upper limit.

[0053] The coefficient of variation (CV value) of the particle size of the silicone particles is calculated by the following formula.

[0054] Coefficient of variation (CV value) (%) of particle size of silicone particles = (ρ / Dn) × 100, where ρ: standard deviation of particle size of silicone particles, and Dn: average particle size of silicone particles.

[0055] When the silicone particles are immersed in toluene at 25°C for 24 hours, the swelling ratio represented by the following formula (L) is preferably 1.1 or more, more preferably 1.2 or more, even more preferably 1.3 or more, even more preferably 1.4 or more, particularly preferably 1.8 or more, preferably 2.5 or less, more preferably 2.3 or less, and even more preferably 2.2 or less. When the swelling ratio is above the lower limit, the silicone particles swell appropriately in the organic solvent, so that the amount of substance that can be retained inside can be increased. Therefore, the reaction efficiency when synthesizing the target molecule can be further improved. When the swelling ratio is below the upper limit, cracking of the silicone particles can be effectively suppressed even during swelling.

[0056] Swelling ratio = L2 / L1 (L) L1: average particle size of silicone particles before immersion in toluene L2: average particle size of silicone particles after immersion in toluene for 24 hours

[0057] More specifically, the swelling ratio is determined as follows.

[0058] 200 random silicone particles are observed under an optical microscope, and the particle size of each silicone particle is determined using imaging software, and the average particle size is calculated. The calculated average particle size is taken as the average particle size (L1) of the silicone particles before immersion in toluene. The average particle size (L1) is the average particle size of the silicone particles in a dry state. 10 mg of silicone particles are immersed in 5 mL of toluene at 25°C for 24 hours. 200 random silicone particles in toluene are observed under an optical microscope, and the particle size of each silicone particle is determined using imaging software, and the average particle size is calculated. The calculated average particle size is taken as the average particle size (L2) of the silicone particles after immersion in toluene for 24 hours. The average particle size (L2) is the average particle size of the silicone particles in a wet state with toluene. The optical microscope may be, for example, a "BX53" manufactured by EVIDENT, and the imaging software may be, for example, "ImageJ" distributed by the National Institute of Health. The particle size of the silicone particles used to determine the average particle sizes (L1) and (L2) refers to the diameter of the particles if they are spherical, and refers to the longest diameter of the particles if they are not spherical.

[0059] Methods for adjusting the swelling ratio within the preferred range include adjusting the content ratio of the polysiloxane moiety and the (meth)acrylic resin moiety in the silicone polymer, adjusting the crosslinking density of the silicone particles, and adjusting the porosity (specific surface area) of the porous silicone particles.

[0060] The shape of the silicone particles is not particularly limited, and may be spherical or a shape other than spherical.

[0061] The aspect ratio of the silicone particles is preferably 1.5 or less, more preferably 1.3 or less. The lower limit of the aspect ratio of the silicone particles is not particularly limited. The aspect ratio of the silicone particles may be 1.0 or more, or may be 1.1 or more. The aspect ratio indicates the major axis / minor axis ratio. The aspect ratio is preferably determined by observing 100 random silicone particles with a scanning electron microscope, defining the longest diameter and the shortest diameter as the major axis and the minor axis, respectively, and calculating the average major axis / minor axis ratio of each silicone particle. The range of the aspect ratio of the silicone particles can be set by appropriately selecting the lower limit value and the upper limit value.

[0062] Other details of the silicone particles will be described below. In this specification, "(meth)acrylic" means either or both of "acrylic" and "methacrylic", "(meth)acryloyl" means either or both of "acryloyl" and "methacryloyl", and "(meth)acrylate" means either or both of "acrylate" and "methacrylate".

[0063] The silicone particles contain a silicone polymer. The silicone polymer has a polysiloxane moiety. That is, the silicone particles contain a silicone polymer having a polysiloxane moiety. The polysiloxane moiety has a structure represented by the following formula (X):

[0064]

[0065] In the above formula (X), R １ is any atom or any group, and R ２ is any atom or any group, and n is an integer of 2 or more. １ and R ２ In the above formula (X), a plurality of R １ In the formula (X), a plurality of R ２ may be the same or different.

[0066] In the above formula (X), R １is preferably a hydrogen atom or an organic group, more preferably a hydrocarbon group, and even more preferably an alkyl group. ２ is preferably a hydrogen atom or an organic group, more preferably a hydrocarbon group, and even more preferably an alkyl group.

[0067] In the above formula (X), R １ When is an alkyl group, the number of carbon atoms in the alkyl group is preferably 1 or more, preferably 10 or less, more preferably 5 or less, and even more preferably 3 or less. In this case, the effects of the present invention can be more effectively exhibited.

[0068] In the above formula (X), R ２ When is an alkyl group, the number of carbon atoms in the alkyl group is preferably 1 or more, preferably 10 or less, more preferably 5 or less, and even more preferably 3 or less. In this case, the effects of the present invention can be more effectively exhibited.

[0069] In the above formula (X), R １ is preferably a methyl group or an ethyl group, and more preferably a methyl group. In this case, the effects of the present invention can be more effectively exhibited.

[0070] In the above formula (X), R ２ is preferably a methyl group or an ethyl group, and more preferably a methyl group. In this case, the effects of the present invention can be more effectively exhibited.

[0071] The silicone particles preferably contain a silicone polymer having a polysiloxane portion and a (meth)acrylic resin portion. In this case, for example, the silicone particles swell moderately in an organic solvent, so that the amount of material that can be retained inside can be increased compared to particles that do not swell. Therefore, the reaction efficiency when synthesizing the target molecule can be further improved. The (meth)acrylic resin portion means a structure represented by the following formula (Y):

[0072]

[0073] In the above formula (Y), R１ is a hydrogen atom or a methyl group, and R ２ is an arbitrary group, and n is an integer of 1 or more. １ In the formula (Y), a plurality of R ２ may be the same or different.

[0074] Examples of methods for producing the silicone polymer having a polysiloxane portion and a (meth)acrylic resin portion include the following methods (1), (2), and (3): (1) a method of radically polymerizing a polymerization component containing a (meth)acryloyl group-containing silicone compound; (2) a method of reacting a polymer of a (meth)acryloyl group-containing monomer with a silicone compound containing a crosslinkable functional group; and (3) a method of introducing an alkoxysilyl group into a polymer of a (meth)acryloyl group-containing monomer, and then introducing a polysiloxane structure by a sol-gel reaction initiated from the alkoxysilyl group.

[0075] <(Meth)acryloyl Group-Containing Silicone Compound (A)> The silicone polymer preferably has a skeleton derived from a (meth)acryloyl group-containing silicone compound (hereinafter sometimes referred to as a (meth)acryloyl group-containing silicone compound (A)). The material for the silicone polymer preferably contains the (meth)acryloyl group-containing silicone compound (A). The (meth)acryloyl group-containing silicone compound (A) refers to a compound having a siloxane moiety and a (meth)acryloyl group. The (meth)acryloyl group-containing silicone compound (A) is preferably a compound having a polysiloxane moiety and a (meth)acryloyl group. Only one type of the (meth)acryloyl group-containing silicone compound (A) may be used, or two or more types may be used in combination.

[0076] From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the polysiloxane moiety of the silicone polymer contains the polysiloxane moiety of the (meth)acryloyl group-containing silicone compound (A).

[0077] In order to more effectively exert the effects of the present invention, it is preferable that the main chain of the silicone polymer contains a skeleton derived from the (meth)acryloyl group-containing silicone compound (A).

[0078] The (meth)acryloyl group-containing silicone compound (A) is preferably a (meth)acryloyl group-containing silicone compound having a (meth)acryloyl group at one or both ends, more preferably a (meth)acryloyl group-containing silicone compound having a (meth)acryloyl group at both ends, and even more preferably a (meth)acryloyl group-containing silicone compound represented by the following formula (A1): In this case, the effects of the present invention can be even more effectively exhibited.

[0079]

[0080] In the above formula (A1), R １ , R ２ , R ３ and R ４ each represents an organic group, R ５ and R ６ Each of R represents a group having a (meth)acryloyl group, and n represents an integer of 1 or more. １ , R ２ , R ３ and R ４ In the above formula (A1), R ５ and R ６ may be the same or different. １ In the formula (A1), a plurality of R ２ may be the same or different.

[0081] In the above formula (A1), R １ , R ２ , R ３ and R ４ is preferably a hydrocarbon group, more preferably an alkyl group, in which case the effects of the present invention can be more effectively exhibited.

[0082] In the above formula (A1), R １ , R ２ , R ３ and R ４ When each of the groups is an alkyl group, the number of carbon atoms in the alkyl group is preferably 1 or more, preferably 10 or less, more preferably 5 or less, and even more preferably 3 or less. In this case, the effects of the present invention can be more effectively exhibited.

[0083] In the above formula (A1), R １ , R ２ , R ３ and R ４ is preferably a methyl group or an ethyl group, and more preferably a methyl group. In this case, the effects of the present invention can be more effectively exhibited.

[0084] In the above formula (A1), n ​​is preferably 4 or greater, more preferably 8 or greater, and preferably 100 or less, more preferably 50 or less. When n is equal to or greater than the above lower limit and equal to or less than the above upper limit, the silicone particles can be imparted with appropriate flexibility and strength, thereby effectively preventing the silicone particles from cracking.

[0085] Commercially available (meth)acryloyl group-containing silicone compounds represented by formula (A1) include "X-22-164," "X-22-164AS," "X-22-164A," "X-22-164B," "X-22-164C," "X-22-164E," and "X-22-2445" manufactured by Shin-Etsu Silicones Co., Ltd., and "FM-7711" and "FM-7721" manufactured by JNC Corporation.

[0086] In 100% by weight of the silicone polymer (100% by weight of the total skeletons possessed by the silicone polymer), the content of skeletons derived from the (meth)acryloyl group-containing silicone compound (A) is preferably 50% by weight or more, more preferably 60% by weight or more, even more preferably 70% by weight or more, preferably 100% by weight or less, more preferably 97% by weight or less, and even more preferably 95% by weight or less.When the content of skeletons derived from the (meth)acryloyl group-containing silicone compound (A) is above the lower limit, the silicone particles can be endowed with appropriate flexibility and appropriate strength, so that the cracking of the silicone particles can be effectively suppressed.When the content of skeletons derived from the (meth)acryloyl group-containing silicone compound (A) is below the upper limit, the amount of substance that can be retained inside can be further increased, so that the reaction efficiency when synthesizing the target molecule can be further improved.

[0087] <Radical Polymerizable Compound (B)> The silicone polymer preferably further has a skeleton derived from a radically polymerizable compound (hereinafter, sometimes referred to as radically polymerizable compound (B)) different from the (meth)acryloyl group-containing silicone compound. The material of the silicone polymer preferably contains the radically polymerizable compound (B). The radically polymerizable compound (B) is different from the (meth)acryloyl group-containing silicone compound (A). The radically polymerizable compound (B) is different from a compound having both a polysiloxane moiety and a (meth)acryloyl group. The radically polymerizable compound (B) may or may not have a polysiloxane moiety. The radically polymerizable compound (B) may or may not have a (meth)acryloyl group. The radically polymerizable compound (B) has a radically polymerizable functional group. Only one type of the radically polymerizable compound (B) may be used, or two or more types may be used in combination.

[0088] In order to more effectively exert the effects of the present invention, the silicone polymer preferably has a skeleton derived from the radically polymerizable compound (B).

[0089] From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the main chain of the silicone polymer contains a skeleton derived from the radical polymerizable compound (B). From the viewpoint of even more effectively exerting the effects of the present invention, it is preferable that the main chain of the silicone polymer contains a skeleton derived from the (meth)acryloyl group-containing silicone compound (A) and a skeleton derived from the radical polymerizable compound (B).

[0090] Examples of the radically polymerizable functional group contained in the radically polymerizable compound (B) include a (meth)acryloyl group, a vinyl group, a styryl group, an allyl group, etc. The radically polymerizable compound (B) may have only one type of radically polymerizable functional group, or two or more types.

[0091] From the viewpoint of more effectively exerting the effects of the present invention, it is preferable that the radically polymerizable functional group possessed by the radically polymerizable compound (B) contains a (meth)acryloyl group. That is, it is preferable that the radically polymerizable compound (B) contains a compound having a (meth)acryloyl group (hereinafter, sometimes referred to as a (meth)acryloyl group-containing compound (B1)). The (meth)acryloyl group-containing compound (B1) is different from a (meth)acryloyl group-containing silicone compound. The (meth)acryloyl group-containing compound (B1) refers to a compound that does not have a polysiloxane moiety and has a (meth)acryloyl group. The (meth)acryloyl group-containing compound (B1) may be used alone or in combination of two or more types.

[0092] Examples of the (meth)acryloyl group-containing compound (B1) include (meth)acrylic acid, (meth)acrylic acid esters, (meth)acrylamide, and (meth)acrylamide derivatives.

[0093] Examples of the (meth)acrylic acid ester include methyl (meth)acrylate, ethyl (meth)acrylate, propyl (meth)acrylate, butyl (meth)acrylate, 2-ethylhexyl (meth)acrylate, lauryl (meth)acrylate, cetyl (meth)acrylate, stearyl (meth)acrylate, cyclohexyl (meth)acrylate, isobornyl (meth)acrylate, benzyl (meth)acrylate, 2-hydroxyethyl (meth)acrylate, 4-hydroxyphenyl (meth)acrylate, glycerol (meth)acrylate, polyoxyethylene (meth)acrylate, glycidyl (meth)acrylate, trifluoromethyl (meth)acrylate, pentafluoroethyl (meth)acrylate, pentaerythritol tetraacrylate, and polytetramethylene glycol diacrylate.

[0094] Examples of the (meth)acrylamide derivatives include N-isopropyl(meth)acrylamide, N-methoxymethyl(meth)acrylamide, and N,N-dimethyl(meth)acrylamide.

[0095] The radical polymerizable compound (B) may or may not contain a radical polymerizable compound having no (meth)acryloyl group. Examples of the radical polymerizable compound having no (meth)acryloyl group include styrenes such as styrene, 4-methylstyrene, α-methylstyrene, chlorostyrene, 4-chloromethylstyrene, bromostyrene, 4-methoxystyrene, 4-tert-butoxystyrene, nitrostyrene, aminostyrene, carboxystyrene, 4-(methoxycarbonyl)styrene, and sodium 4-styrenesulfonate; divinylbenzenes such as divinylbenzene, 1,2-di(4-vinylphenyl)methane, and 1,2-di(4-vinylphenyl)ethane; vinyl compounds having a lactam structure such as N-vinyl-2-pyrrolidone and N-vinyl-ε-caprolactam; (meth)acrylonitrile and (meth)acrylonitrile derivatives.

[0096] The radical polymerizable compound (B) preferably contains a radical polymerizable compound having a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group. In this case, the silicone polymer can be reacted with a substance held inside the silicone particle. That is, the functional group (reactive group) can be reacted with the reactive group of the substance to bond the silicone polymer to the substance. Therefore, for example, the silicone particle can be suitably used as a support for solid-phase synthesis of peptides or nucleic acids. In this case, the skeleton derived from the radical polymerizable compound (B) in the solid-phase synthesis support can serve as an introduction point for a linker. The radical polymerizable compound (B) having a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group may be a (meth)acryloyl group-containing compound (B1) or a radical polymerizable compound other than a (meth)acryloyl group-containing compound (B1).

[0097] The (meth)acryloyl group-containing compound (B1) preferably has a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group. In this case, the silicone polymer can be reacted with a substance held inside the silicone particle. That is, the functional group (reactive group) can be reacted with the reactive group of the substance to bond the silicone polymer with the substance. Therefore, for example, the silicone particle can be suitably used as a support for solid-phase synthesis of peptides or nucleic acids. In this case, the skeleton derived from the (meth)acryloyl group-containing compound (B1) in the solid-phase synthesis support can serve as an introduction point for a linker.

[0098] In 100% by weight of the silicone polymer (100% by weight of the total skeletons possessed by the silicone polymer), the content of skeletons derived from the radical polymerizable compound (B) is preferably 1% by weight or more, more preferably 2% by weight or more, even more preferably 3% by weight or more, preferably 20% by weight or less, more preferably 15% by weight or less, and even more preferably 10% by weight or less. When the content of skeletons derived from the radical polymerizable compound (B) is above the lower limit, the amount of substance that can be retained inside can be further increased, thereby further improving the reaction efficiency when synthesizing the target molecule. When the content of skeletons derived from the radical polymerizable compound (B) is below the upper limit, cracking of the silicone particles can be effectively suppressed.

[0099] <Other Details of Silicone Particles> It is preferable that a carboxyl group, an amino group, a hydroxyl group, a nitrile group, an acetoxy group, or a halogeno group is present on the surface of the silicone particles. In this case, the silicone polymer can react with the substance held inside the silicone particles. That is, the functional group (reactive group) can react with the reactive group of the substance and bond to it. Therefore, the reaction efficiency when synthesizing the target molecule can be further improved.

[0100] The type of functional group present on the surface of the silicone particles can be identified by Fourier transform infrared spectroscopy.

[0101] In the silicone polymer, the content of the skeleton derived from the (meth)acryloyl group-containing silicone compound (A) in the total of 100% by weight of the skeleton derived from the (meth)acryloyl group-containing silicone compound (A) and the skeleton derived from the radical polymerizable compound (B) is preferably 50% by weight or more, more preferably 55% by weight or more, even more preferably 60% by weight or more, preferably 99% by weight or less, more preferably 97% by weight or less, and even more preferably 95% by weight or less.When the content of the skeleton derived from the (meth)acryloyl group-containing silicone compound (A) is above the lower limit, the silicone particles can be imparted with appropriate flexibility and appropriate strength, so that the cracking of the silicone particles can be effectively suppressed.When the content of the skeleton derived from the (meth)acryloyl group-containing silicone compound (A) is below the upper limit, the amount of substance that can be retained inside the silicone particles can be further increased, so that the reaction efficiency when synthesizing the target molecule can be further improved.

[0102] The content of the skeleton derived from the (meth)acryloyl group-containing silicone compound (A) and the content of the skeleton derived from the radical polymerizable compound (B) can be measured, for example, by NMR (nuclear magnetic resonance).

[0103] The silicone polymer may have a skeleton derived from a compound other than both the (meth)acryloyl group-containing silicone compound (A) and the radically polymerizable compound (B), as long as it does not contradict the object of the present invention.

[0104] The content of the silicone polymer in 100% by weight of the silicone particles is preferably 95% by weight or more, more preferably 98% by weight or more, even more preferably 99% by weight or more, and most preferably 100% by weight (total amount).In addition, the content of the silicone polymer in 100% by weight of the silicone particles may be 100% by weight or less, may be less than 100% by weight, or may be 99% by weight or less.The range of the content of the silicone polymer can be set by appropriately selecting the lower limit and the upper limit.

[0105] (Method for producing porous silicone particles) The method for producing the porous silicone particles described above is described below. The method for producing the porous silicone particles is preferably a method for producing porous silicone particles containing a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety.

[0106] The method for producing the porous silicone particles preferably comprises the steps of: 1) obtaining a mixture containing a polymerization component and a porogen; 2) dispersing the mixture in a dispersion medium to obtain a dispersion; 3) polymerizing the polymerization component in the dispersion to obtain particles; and 4) removing the porogen from the particles.

[0107] <Step of Obtaining a Mixture> The mixture can be obtained by mixing a polymerization component and a porogen. The polymerization component is a material for the porous silicone particles and the silicone polymer. The porogen is a pore-forming agent. The mixture is preferably a liquid mixture containing the polymerization component and the porogen.

[0108] The polymerization component preferably contains a (meth)acryloyl group-containing silicone compound (A). The (meth)acryloyl group-containing silicone compound (A) described above can be used as the (meth)acryloyl group-containing silicone compound (A). In the method for producing porous silicone particles, the preferred configuration and other features of the (meth)acryloyl group-containing silicone compound (A) are the same as those described in the section <(meth)acryloyl group-containing silicone compound (A)> above.

[0109] The polymerization component preferably contains a radical polymerizable compound (B) different from the (meth)acryloyl group-containing silicone compound. The radical polymerizable compound (B) described above can be used as the radical polymerizable compound (B). In the method for producing porous silicone particles, the preferred configuration of the radical polymerizable compound (B) is the same as the configuration described in the section <Radical polymerizable compound (B)> above.

[0110] Examples of the porogen include hydrocarbons and alcohols. Examples of the hydrocarbons include aliphatic hydrocarbons and aromatic hydrocarbons. Examples of the aliphatic hydrocarbons include saturated aliphatic hydrocarbons and unsaturated aliphatic hydrocarbons. Examples of the aliphatic hydrocarbons include n-hexane, n-pentane, n-heptane, n-octane, n-isooctane, n-decane, n-dodecane, and n-hexadecane. Examples of the aromatic hydrocarbons include toluene, benzyl alcohol, and dibutyl phthalate. Examples of the alcohols include aliphatic alcohols, more specifically, cellosolves such as 1-hexanol, 2-ethylhexanol, 1-pentanol, 1-octanol, 2-octanol, cyclohexanol, methyl cellosolve, ethyl cellosolve, and butyl cellosolve; glycol esters such as ethylene glycol monomethyl ether acetate and propylene glycol monomethyl ether acetate; and glycols such as polyethylene glycol and polypropylene glycol. The porogens may be used alone or in combination of two or more.

[0111] The porogen preferably contains the aliphatic hydrocarbon, more preferably contains the aliphatic hydrocarbon and an aromatic hydrocarbon, and even more preferably contains the aliphatic hydrocarbon, the aromatic hydrocarbon, and the alcohol. In this case, the average pore size and specific surface area of ​​the porous silicone particles can be easily adjusted to fall within the preferred ranges described above. The aliphatic hydrocarbon is preferably an aliphatic hydrocarbon having 5 to 16 carbon atoms, and more preferably n-hexane, n-pentane, n-heptane, n-octane, n-isooctane, n-decane, n-dodecane, or n-hexadecane.

[0112] The mixture may contain a radical polymerization initiator. The step of obtaining the mixture may be a step of obtaining a mixture containing the polymerization components, a porogen, and a radical polymerization initiator.

[0113] In the mixture, the content of the porogen per 100 parts by weight of the polymerization components is preferably at least 1 part by weight, more preferably at least 5 parts by weight, and preferably not more than 60 parts by weight, more preferably not more than 50 parts by weight. When the content of the porogen is at least the above lower limit and not more than the above upper limit, it becomes easier to adjust the average pore diameter and specific surface area of ​​the obtained porous silicone particles to fall within the preferred ranges described above.

[0114] <Step of Obtaining Dispersion> The dispersion can be obtained by dispersing the mixture in a dispersion medium.

[0115] Examples of the dispersion medium include solutions containing water, methanol, and the like. Only one type of dispersion medium may be used, or two or more types may be used in combination. The dispersion medium may be a solution containing water, a solution containing methanol, or a solution containing water and methanol. The dispersion medium may also contain a dispersant such as polyvinyl alcohol or other additives.

[0116] In the dispersion, the content of the mixture relative to 100 parts by weight of the dispersion medium is preferably at least 1 part by weight, more preferably at least 10 parts by weight, and preferably not more than 50 parts by weight, more preferably not more than 40 parts by weight. When the content of the mixture is at least the above-mentioned lower limit and is not more than the above-mentioned upper limit, it becomes easier to adjust the average pore size and specific surface area of ​​the obtained porous silicone particles to fall within the preferred ranges described above.

[0117] <Step of Obtaining Particles> The particles can be obtained by polymerizing the polymerization components in the dispersion.

[0118] The method for polymerizing the polymerization components in the dispersion is not particularly limited as long as it can polymerize the polymerization components. The method for polymerizing the polymerization components in the dispersion may be a method of heating the dispersion or a method of irradiating the dispersion with light.

[0119] From the viewpoint of increasing the polymerization efficiency, the method of polymerizing the polymerization components in the dispersion is preferably a method of heating the dispersion.

[0120] <Step of Removing Porogen> Particles having a porous structure (porous silicone particles) can be obtained by removing the porogen from the particles obtained.

[0121] The porogen can be removed by washing the particles with an organic solvent that dissolves the porogen, etc. The organic solvent that dissolves the porogen can be appropriately selected depending on the type of porogen.

[0122] (Method for Producing Target Molecule) A target molecule can be produced using the silicone particles described above. The method for producing a target molecule according to the present invention comprises a step of synthesizing a target molecule on silicone particles. The method for producing a target molecule according to the present invention is a method for producing a target molecule by a solid phase method. The silicone particles are a support for solid phase synthesis. The method for producing a target molecule according to the present invention can employ a conventionally known method, except that the silicone particles are used as a support for solid phase synthesis.

[0123] Examples of the target molecule include peptides and nucleic acids.

[0124] The target molecule is preferably a peptide or a nucleic acid, more preferably a peptide. The method for producing the target molecule is preferably a method for producing a peptide or a nucleic acid, more preferably a method for producing a peptide.

[0125] Hereinafter, the method for producing a target molecule of the present invention will be described using the case where the target molecule is a peptide, that is, the case where a peptide is produced, as an example.

[0126] <Method for Producing Peptides> Peptides can be produced by conventionally known methods, except that the silicone particles described above are used as a support for solid-phase synthesis.

[0127] The method for producing the peptide preferably comprises a peptide synthesis step of synthesizing peptide on the silicone particles, and a peptide separation step of separating the synthesized peptide.However, if the silicone particles do not have a linker structure that serves as the starting point of solid-phase peptide synthesis, the method for producing the peptide may also comprise a linker introduction step, prior to the peptide synthesis step, of introducing a linker into the silicone particles to obtain silicone particles with a linker introduced therein.In addition, if the silicone particles have a linker structure that serves as the starting point of solid-phase peptide synthesis, the method for producing the peptide does not need to comprise the linker introduction step.

[0128] <<Linker Introduction Step>> In the linker introduction step, a linker is introduced into the silicone particles to obtain silicone particles with a linker introduced therein. As a result, a linker structure that serves as a starting point for solid-phase peptide synthesis is formed on the silicone particles. Examples of the linker structure include linker structures of commonly known resins for solid-phase synthesis, such as Wang resin, chlorotrityl resin, Rink Amide resin, and Sieber resin.

[0129] <<Peptide Synthesis Step>> In the peptide synthesis step, peptides are synthesized on the silicone particles. In the peptide synthesis step, peptides are extended on the silicone particles. The silicone particles have a linker structure.

[0130] The peptide synthesis process preferably comprises the following steps (A), (C), (D), and (F) in this order, more preferably the following steps (A), (C), (D), (F), and (G) in this order, and even more preferably the following steps (A), (B), (C), (D), (E), (F), and (G) in this order.

[0131] (A) A step of obtaining a silicone particle-amino acid complex in which the silicone particle (silicone particle having a linker structure) is bound to an amino acid having a protecting group.

[0132] (B) A washing step for washing out the reaction system containing the silicone particle-amino acid complex.

[0133] (C) A deprotection step of contacting the silicone particle-amino acid complex with a deprotection agent.

[0134] (D) A step of reacting the amino group of the silicone particle-amino acid complex produced by deprotection with the carboxy group of an amino acid having a protecting group to obtain a silicone particle-peptide complex.

[0135] (E) A washing step for washing out the reaction system containing the silicone particle-peptide complex.

[0136] (F) A deprotection step of contacting the silicone particle-peptide complex with a deprotecting agent.

[0137] (G) A step of reacting the amino group of the silicone particle-peptide complex produced by deprotection with the carboxy group of an amino acid having a protecting group to obtain a silicone particle-peptide complex with an extended peptide chain.

[0138] Step (A): In step (A), it is preferable to obtain a silicone particle-amino acid complex by reacting the silicone particles (silicone particles having a linker structure) with an amino acid having a protecting group. The silicone particle-amino acid complex is an amino acid-bonded silicone particle in which an amino acid is bonded to a silicone particle. For example, a silicone particle-amino acid complex is obtained by reacting a functional group (reactive group) present on the surface of the silicone particle with a carboxy group of the amino acid.

[0139] Examples of the protecting group include an Fmoc group and a Boc group.

[0140] Step (B): In step (B), the reaction system containing the silicone particle-amino acid complex is preferably washed out with an organic solvent. Any conventional organic solvent used in solid-phase peptide synthesis can be used as the organic solvent.

[0141] Examples of the organic solvent include N,N-dimethylformamide (DMF), methylene chloride, N-methylpyrrolidone, dimethylacetamide, etc. The organic solvents may be used alone or in combination of two or more.

[0142] Step (C): In step (C), the silicone particle-amino acid complex is contacted with a deprotecting agent. A conventionally known deprotecting agent used in solid-phase peptide synthesis can be used as the deprotecting agent. The deprotecting agent is appropriately selected depending on the type of protecting group. For example, when the protecting group is an Fmoc group, a weak base such as piperidine can be used as the deprotecting agent, and when the protecting group is a Boc group, a strong acid such as trifluoroacetic acid (TFA) can be used as the deprotecting agent.

[0143] Step (D): In step (D), the amino group of the silicone particle-amino acid complex formed by deprotection reacts with the carboxy group of the amino acid having a protecting group to obtain a silicone particle-peptide complex.

[0144] Examples of the protecting group include an Fmoc group and a Boc group.

[0145] Step (E): In step (E), the reaction system containing the silicone particle-peptide complex is preferably washed out with an organic solvent. Any conventional organic solvent used in solid-phase peptide synthesis can be used as the organic solvent.

[0146] Examples of the organic solvent that can be used in step (E) include the organic solvents described in the section for step (B).

[0147] Step (F): In step (F), the silicone particle-peptide complex is brought into contact with a deprotecting agent. Any known deprotecting agent used in solid-phase peptide synthesis can be used as the deprotecting agent. The deprotecting agent is appropriately selected depending on the type of protecting group. Examples of the deprotecting agent that can be used in step (F) include the deprotecting agents described in the section for step (C).

[0148] Step (G): In step (G), the amino group of the silicone particle-peptide complex produced by deprotection reacts with the carboxy group of the amino acid having a protecting group to obtain a silicone particle-peptide complex with an elongated peptide chain. The peptide chain of the silicone particle-peptide complex obtained in step (G) has one more amino acid residue than the peptide chain of the silicone particle-peptide complex used in step (F).

[0149] In the method for producing the peptide, it is preferable to further repeat steps (E), (F), and (G) after step (G). The number of times steps (E), (F), and (G) are further repeated after step (G) is appropriately changed depending on the length of the desired peptide chain.

[0150] <<Peptide Separation Step>> In the peptide separation step, the synthesized peptide is separated. More specifically, in the peptide separation step, the synthesized peptide is separated from the silicone particle portion of the silicone particle-peptide complex. In the peptide separation step, the silicone particle portion and the peptide portion of the silicone particle-peptide complex are cleaved to separate the peptide. In this way, the peptide, which is the target molecule, can be obtained.

[0151] The present invention will be specifically described below by way of examples and comparative examples, but the present invention is not limited to the following examples.

[0152] The following monomers (polymerization components) were prepared.

[0153] ((Meth)acryloyl group-containing silicone compound (A)) (a1) Both-end methacrylic-modified silicone compound 1 ("X-22-164A" manufactured by Shin-Etsu Chemical Co., Ltd.) (a2) Both-end methacrylic-modified silicone compound 2 ("X-22-164AS" manufactured by Shin-Etsu Chemical Co., Ltd.) (a3) ​​Both-end methacrylic-modified silicone compound 3 ("X-22-164" manufactured by Shin-Etsu Chemical Co., Ltd.) (a4) Both-end acrylic-modified silicone compound ("X-22-2445" manufactured by Shin-Etsu Chemical Co., Ltd.)

[0154] (Radical polymerizable compound (B)) (b1) methacrylic acid (b2) 2-hydroxyethyl methacrylate (b3) pentaerythritol tetraacrylate ("A-TMMT" manufactured by Shin-Nakamura Chemical Co., Ltd.) (b4) polytetramethylene glycol diacrylate ("Light Acrylate PTMGA-250" manufactured by Kyoeisha Chemical Co., Ltd.)

[0155] (Other Polymerization Components) (c1) Tetraethoxysilane (c2) Methyltrimethoxysilane

[0156] The following compounds were prepared as porogen materials.

[0157] (p1) Toluene (p2) Dibutyl phthalate (p3) 2-ethylhexanol (p4) Hexadecane

[0158] Example 1 <Step of Obtaining a Mixture (Mixed Liquid)> The materials listed in the table below were mixed to prepare a polymerization component and a porogen, respectively. After mixing the polymerization component and the porogen to obtain the weight ratios listed in the table below, 7 parts by weight of a radical polymerization initiator (tert-butyl-2-ethyloxyhexanoate, "Perbutyl O" manufactured by NOF Corporation) was added per 100 parts by weight of the polymerization component, and the mixture was stirred until homogeneous, to obtain a mixed liquid.

[0159] <Step of Obtaining Dispersion Liquid> Next, a dispersion medium was prepared by mixing 500 parts by weight of pure water, 13 parts by weight of polyvinyl alcohol (degree of polymerization: approximately 2000, degree of saponification: 86.5 to 89 mol%, "GOHSENOL GH-20" manufactured by Mitsubishi Chemical Corporation), and 0.1 parts by weight of sodium nitrite. 10 parts by weight of the mixture was mixed with 100 parts by weight of the dispersion medium and stirred to obtain a dispersion liquid.

[0160] <Step of Obtaining Particles> The obtained dispersion was heated at 90° C. for 9 hours to carry out a radical polymerization reaction, thereby obtaining particles.

[0161] <Step of Removing Porogen> The resulting particles were washed three times each with hot water and acetone, and then classified to recover the porous silicone particles. The resulting porous silicone particles comprise a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety.

[0162] Examples 2 and 3 Porous silicone particles were obtained in the same manner as in Example 1, except that the type and content of the polymerization component, the type and content of the porogen, and the mixing ratio (weight ratio) of the polymerization component to the porogen were changed as shown in the table below. The obtained porous silicone particles contain a silicone polymer having a polysiloxane moiety and a (meth)acrylic resin moiety.

[0163] Example 4 Solid silicone particles were obtained in the same manner as in Example 1, except that the types and amounts of the polymerization components were changed as shown in the table below and no porogen was used.

[0164] (Comparative Examples 1 and 2) Porous acrylic resin particles were obtained in the same manner as in Example 1, except that the type and content of the polymerization component, the type and content of the porogen, and the mixing ratio (weight ratio) of the polymerization component to the porogen were changed as shown in the table below.

[0165] Comparative Example 3: A monomer solution was prepared by adding the polymerization components obtained by mixing the materials listed in the table below to 1,000 parts by weight of ethanol and mixing them. Next, a mixed solution was prepared containing 2,000 parts by weight of ethanol, 200 parts by weight of water, 100 parts by weight of 30 wt % aqueous ammonia, 2 parts by weight of potassium chloride, and 2 parts by weight of hexadecyltrimethylammonium bromide, and the monomer solution was slowly added thereto to obtain porous silica particles by a sol-gel reaction. The obtained porous silica particles were washed with water and then collected.

[0166] (Evaluation) The particles obtained in the examples and comparative examples were evaluated as follows, and the results are shown in the table below.

[0167] (1) Average particle size and coefficient of variation (CV value) of particle size The average particle size and coefficient of variation (CV value) of particle size of the obtained particles were measured by the method described above. Note that the scanning electron microscope used was "Regulus-8220" manufactured by Hitachi High-Technologies Corporation, and the imaging software used was "ImageJ" distributed by the National Institute of Health.

[0168] (2) Compressive Elastic Modulus (10% K Value) The compressive elastic modulus (10% K value) of the obtained particles was measured by the method described above. Note that a Fischerscope H-100 manufactured by Fischer was used as the microcompression tester.

[0169] (3) Average pore size and specific surface area The average pore size and specific surface area of ​​the obtained particles were measured by the above-mentioned method. Note that a gas adsorption pore size distribution measuring device, "Nova Touch 4LX" manufactured by Quantachrome, was used.

[0170] (4) Swelling ratio The swelling ratio of the obtained particles was measured by the method described above. Note that the optical microscope used was "BX53" manufactured by EVIDENT, and the imaging software used was "ImageJ" distributed by the National Institute of Health.

[0171] (5) Reactive groups (functional groups) present on the particle surface The reactive groups (functional groups) present on the particle surface were identified by Fourier transform infrared spectroscopy. Specifically, they were measured by the potassium bromide (KBr) tablet method.

[0172] (6) Amount of substance retained inside the particles: A fluorescent substance (Nile Red, manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was mixed with toluene to a concentration of 30 μg / mL to prepare a fluorescent substance-containing toluene solution. 10 mL of the fluorescent substance-containing toluene solution was placed in a beaker, and then 10 mg of particles were added and allowed to stand for 10 minutes. Next, using a magnetic stirrer and a stirring bar, the solution containing the fluorescent substance-containing toluene solution and particles was stirred at 200 rpm for 10 minutes. After stirring, the solution was allowed to stand for 10 minutes. Next, the cross-sections of the particles were photographed using a confocal laser microscope (Yokogawa Electric Corporation's "CellVoyager CQ1"). In the obtained micrographs, the percentage (X) of the area with a brightness value of 30 or higher was calculated based on the 100% cross-sectional area of ​​the particles.

[0173] [Criteria for determining the amount of substance retained inside particles] A: The ratio (X) is 50% or more B: The ratio (X) is less than 50%

[0174] (7) Cracking After the test in "(6) Amount of substance held inside particles" above, 100 randomly selected particles were observed under an optical microscope (EVIDENT's "BX53").

[0175] [Crack evaluation criteria] A: No cracks in particles B: Cracks in particles (cracked particles present)

[0176] (8) Reaction Efficiency (Peptide Elongation Efficiency) (8-1) Linker Introduction Step: 0.4 parts by weight of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and 2 parts by weight of 4-hydroxybenzaldehyde were dissolved in 200 parts by weight of dimethylformamide (DMF), and then 10 parts by weight of the resulting particles were added and stirred at 40°C for 6 hours. After stirring, the particles were washed with DMF and water, and then vacuum-dried. Five parts by weight of the vacuum-dried particles were suspended in 100 parts by weight of tetrahydrofuran (THF), and 0.1 parts by weight of sodium borohydride was added at 0°C. The mixture was stirred for 20 hours while heating to 70°C. The particles were then washed with THF and water, and then vacuum-dried. The resulting particles were analyzed using a Fourier transform infrared spectrometer, confirming that particles with a Wang resin linker structure had been introduced. For the particles of Example 4 and Comparative Example 3, the same procedure was carried out using 4-carboxybenzaldehyde instead of 4-hydroxybenzaldehyde.

[0177] (8-2) Peptide Synthesis Step: A glass column was packed with 1 part by weight of particles incorporating a Wang resin linker structure, and the procedure of washing with 5 parts by weight of DMF was repeated three times. Next, a DMF solution was prepared by dissolving 0.4 parts by weight of Fmoc-aspartic acid (Obu), 0.1 parts by weight of N,N-diisopropylcarbodiimide, and 0.1 parts by weight of 1-hydroxybenzotriazole in 5 parts by weight of DMF. This DMF solution was added to the glass column packed with particles and allowed to stand at 25°C for 2 hours to carry out the introduction reaction of Fmoc-aspartic acid (Obu). After the reaction, the particles were washed with DMF three times. Next, 5 parts by weight of a piperidine / DMF (20% v / v) solution was added, and after standing for 20 minutes, the solution was removed from the column, and the particles were washed with DMF. Next, the same procedure was carried out using Fmoc-glycine-OH to elongate the peptide with Fmoc-glycine-OH. Furthermore, the peptide was elongated with Fmoc-arginine(pbf)-OH by carrying out the same procedure using Fmoc-arginine(pbf)-OH. After washing with DMF, methanol, and dichloromethane in that order, the product was dried under reduced pressure for 12 hours.

[0178] (8-3) Peptide Separation Step Next, the particles were stirred for 2 hours in a mixed solution of trifluoroacetic acid and dichloromethane (9 / 1, v / v), followed by suction filtration and the same procedure again. Trifluoroacetic acid was removed from the obtained filtrate using an evaporator, and the residue was washed with methanol. Finally, pure water was added, and the mixture was freeze-dried for 2 days to obtain peptides. The obtained peptides were analyzed by high-performance liquid chromatography, and the yield of the final product, arginine-glycine-aspartic acid (Obu), was calculated. The calculated yield was used as the peptide elongation efficiency.

[0179] Details and results are shown in Tables 1 to 4 below.

[0180]

[0181]

[0182]

[0183]

[0184] 1...Silicone particles

Claims

1. A method for producing a target molecule, comprising a step of synthesizing the target molecule on silicone particles.

2. The method for producing a target molecule according to claim 1, wherein the silicone particles are porous silicone particles.

3. The method for producing a target molecule according to claim 2, wherein the porous silicone particles have an average pore size of 2 nm or more and 300 nm or less.

4. The compressive modulus of the silicone particles when compressed by 10% is 500 N / mm ２ The method for producing a target molecule according to any one of claims 1 to 3, wherein:

5. The method for producing a target molecule according to any one of claims 1 to 4, wherein when the silicone particles are immersed in toluene at 25°C for 24 hours, the swelling ratio, represented by the following formula (L), is 1.1 or more and 2.5 or less: Swelling ratio = L2 / L1 (L), where L1 is the average particle size of the silicone particles before immersion in toluene, and L2 is the average particle size of the silicone particles after immersion in toluene for 24 hours.

6. The method for producing a target molecule according to any one of claims 1 to 5, wherein a carboxy group, amino group, hydroxy group, nitrile group, acetoxy group, or halogeno group is present on the surface of the silicone particle.

7. The method for producing a target molecule according to any one of claims 1 to 6, wherein the silicone particles contain a silicone polymer having a polysiloxane portion and a (meth)acrylic resin portion.

8. The method for producing a target molecule according to claim 7, wherein the silicone polymer has a skeleton derived from a (meth)acryloyl group-containing silicone compound (A).

9. The method for producing a target molecule according to claim 8, wherein the silicone polymer further has a skeleton derived from a radically polymerizable compound (B) different from the (meth)acryloyl group-containing silicone compound.

10. The method for producing a target molecule according to claim 9, wherein the radical polymerizable compound (B) includes a (meth)acryloyl group-containing compound (B1).

11. The method for producing a target molecule according to claim 9 or 10, wherein the radical polymerizable compound (B) comprises a radical polymerizable compound having a carboxy group, an amino group, a hydroxy group, a nitrile group, an acetoxy group, or a halogeno group.

12. The method for producing a target molecule according to any one of claims 1 to 11, wherein the average particle size of the silicone particles is 10 μm or more and 500 μm or less.

13. The specific surface area of ​​the silicone particles is 1 m ２ / g or more 200m ２ The method for producing a target molecule according to any one of claims 1 to 12, wherein the total amount of the target molecule is 1 / g or less.

14. The method for producing a target molecule according to any one of claims 1 to 13, wherein the target molecule is a peptide or a nucleic acid.

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