Separation and purification method for medium-sized molecule pharmaceutical and separation and purification system for medium-sized molecule pharmaceutical

Abstract

The present invention provides a separation and purification method and a separation and purification system that make it possible to purify a highly pure medium-sized molecule pharmaceutical with a high yield, even when applied to production on a large scale.　A separation and purification method for icatibant according to one embodiment comprises: a primary processing step for supplying a processing target liquid containing icatibant and a plurality of impurities to a column which has been filled with a separation agent X, thereby obtaining a primary processing liquid in which an α component has been removed from the processing target liquid; and a secondary processing step for supplying the primary processing liquid to a plurality of filling parts which have been filled with a separation agent Y, thereby performing a simulated moving bed method for continuously separating the icatibant.　The α component is an impurity among the plurality of impurities in the processing target liquid which, in a chromatogram of the processing target liquid obtained in the primary processing step, has the smallest overlap with the peak area of the icatibant.

Description

Method for separating and purifying medium-molecular-weight drugs and system for separating and purifying medium-molecular-weight drugs

[0001] The present invention relates to a method for separating and purifying medium-sized molecular weight drugs and a system for separating and purifying medium-sized molecular weight drugs. This application claims priority based on Japanese Patent Application No. 2023-152292, filed with the Japan Patent Office on September 20, 2023, the contents of which are incorporated herein by reference.

[0002] Medium-molecule drugs are drugs containing compounds with a molecular weight of 500 to 10,000, and are positioned between small-molecule drugs and large-molecule drugs. Medium-molecule drugs typically include peptide drugs and nucleic acid drugs. Peptide drugs are drugs whose main ingredient is a peptide with multiple amino acid residues. Among the many peptide drugs, next-generation peptide drugs with non-natural amino acid residues include glucagon-like peptide 1 (GLP-1) such as tirzepatide and icatibant.

[0003] Icatibant is a peptide consisting of a 10-amino acid sequence. Because it is a specific antagonist of the bradykinin B2 receptor, it is used to treat attacks of hereditary angioedema (HAE). Icatibant is synthesized by solid-phase peptide synthesis, but during this synthesis, several impurities, such as aggregated peptides, are generated. Therefore, the purity of icatibant is increased by removing these impurities from the crude icatibant obtained by the synthesis reaction. For example, Patent Document 1 proposes a method for separating and purifying icatibant using reverse-phase chromatography.

[0004] EP 4083053

[0005] However, the method of Patent Document 1 only achieves a recovery rate of approximately 80-85% for icatibant, resulting in significant losses when applied to large-scale production. Because therapeutic drugs containing medium-sized molecule drugs such as icatibant are expensive, improving the recovery rate is desirable to reduce costs. However, further increasing the recovery rate of medium-sized molecule drugs such as icatibant results in a decrease in purity. This is because the crude product to be purified contains multiple impurities with chemical properties similar to those of the medium-sized molecule drug.

[0006] For the above reasons, it is technically very difficult to increase the recovery rate by even 1% while maintaining high purity in the separation and purification of medium-molecule drugs such as icatibant.

[0007] An object of the present invention is to provide a separation and purification method and a separation and purification system that can purify high-purity medium-molecule pharmaceuticals with a high recovery rate even when applied to large-scale production.

[0008] The inventors of the present invention conducted research into how to achieve high purity and high recovery rates in the purification of medium-molecule pharmaceuticals, and came up with the idea of ​​a two-stage purification process that includes primary and secondary treatment. In the primary treatment, the target substance is recovered to the maximum extent by removing components that are easy to separate. In the secondary treatment, precision separation using a simulated moving bed is used. This two-stage purification process that includes primary and secondary treatment can provide a separation and purification method and system that can achieve both high purity and high recovery rates even in large-scale production.

[0009] The present invention encompasses the following embodiments [1] to

[14] . [1] A method for separating and purifying a medium-sized molecular drug, which is a peptide or nucleic acid drug containing a compound with a molecular weight of 500 to 10,000, comprising: a primary treatment step of supplying a treatment target liquid containing a medium-sized molecular drug and multiple impurities to a column packed with separating agent X to obtain a primary treatment liquid from the treatment target liquid, in which the α component described below has been removed; and a secondary treatment step of performing a simulated moving bed method to continuously separate the medium-sized molecular drug by supplying the primary treatment liquid to multiple packing sections packed with separating agent Y, wherein the medium-sized molecular drug is a peptide or nucleic acid drug containing a compound with a molecular weight of 500 to 10,000. The α component is the impurity, among the multiple impurities in the treatment target liquid, that has the smallest overlap with the peak area of ​​the medium-sized molecular drug in the chromatogram of the treatment target liquid obtained in the primary treatment step. [2] The method for separating and purifying the medium-sized molecular drug described in [1], in which the medium-sized molecular drug is icatibant. [3] The separation and purification method according to [1] or [2], further comprising the following α component selection step before the primary treatment step, wherein the α component selection step comprises the steps of: determining a first peak corresponding to the medium molecular weight drug in a chromatogram obtained when the treatment target liquid is supplied to a column packed with the separating agent X; determining a second peak corresponding to an impurity that passed through the separating agent X before the medium molecular weight drug among the plurality of impurities in the chromatogram; determining a third peak corresponding to an impurity that passed through the separating agent X after the medium molecular weight drug; and selecting, from the second and third peaks, the impurity corresponding to the peak that overlaps with the first peak as the α component. [4] The separation and purification method according to any of [1] to [3], wherein, in the chromatogram of the primary treatment step, a fraction of the treatment target liquid from the starting point of the first peak corresponding to the medium molecular weight drug is retained to obtain a primary treatment liquid, and the impurities that flow out before obtaining the primary treatment liquid are removed.[5] The separation and purification method according to any one of [1] to [4], wherein a fraction of the liquid to be treated corresponding to a peak formed by the medium-molecule drug and a peak of an α-component in the chromatogram of the primary treatment step is retained to obtain a primary treatment liquid, and the α-component that flows out after obtaining the primary treatment liquid is removed. [6] The separation and purification method according to any one of [1] to [5], wherein the separating agent X is a synthetic adsorbent. [7] The separation and purification method according to any one of [1] to [6], wherein the secondary treatment step comprises: a supply / withdrawal step of supplying the primary treatment liquid and an eluent to at least two or more different packing sections among the plurality of packing sections, respectively, and withdrawing a separation liquid from each packing section to which the primary treatment liquid and the eluent have been supplied, respectively; and a circulation step of circulating the primary treatment liquid and the eluent among the plurality of packing sections in the same flow direction as in the supply / withdrawal step, without additionally supplying the primary treatment liquid and the eluent to the plurality of packing sections. [8] The separation and purification method according to any one of [1] to [7], wherein the separating agent Y is a synthetic adsorbent. [9] In the secondary treatment step, the space velocity is 0.5 h －１ ~10 hours －１

[10] The separation and purification method according to any one of [1] to [9], wherein the primary treatment liquid is supplied to at least one of the plurality of packing sections under the condition of

[10] .

[11] The separation and purification method according to any one of [1] to [9], wherein the secondary treatment step is performed by adjusting the pH of the eluent to 7.0 or less.

[0010]

[11] A system for separating and purifying medium-sized molecular pharmaceuticals, comprising: a primary treatment device that supplies a treatment target liquid containing a medium-sized molecular pharmaceutical and multiple impurities to a column packed with separating agent X to obtain a primary treatment liquid from the treatment target liquid, from which at least a portion of the α-component described below has been removed; and a simulated moving bed apparatus that performs a simulated moving bed method to continuously separate the medium-sized molecular pharmaceuticals by supplying the primary treatment liquid to multiple packing sections packed with separating agent Y, wherein the medium-sized molecular pharmaceuticals are peptide or nucleic acid pharmaceuticals containing compounds with molecular weights of 500 to 10,000; the α-component is the impurity, among the multiple impurities in the treatment target liquid, that has the smallest overlap with the peak area of ​​the icatibant in the chromatogram of the treatment target liquid obtained by the primary treatment device.

[12] The separation and purification system according to

[11] , wherein the separating agent X is a synthetic adsorbent.

[13] The separation and purification system according to

[11] or

[12] , wherein the simulated moving bed apparatus has a supply section for separately supplying the primary treatment liquid and the eluent to each of the plurality of packed sections, and a withdrawal section for withdrawing the separated liquid from each of the plurality of packed sections, and a circulation path satisfying the following conditions 1 and 2 is formed in the simulated moving bed apparatus: Condition 1: The primary treatment liquid and the eluent are not additionally supplied to the plurality of packed sections. Condition 2: The primary treatment liquid and the eluent are circulated between the plurality of packed sections in a direction from the supply section toward the withdrawal section.

[14] The separation and purification system according to any of

[11] to

[13] , wherein the separating agent Y is a synthetic adsorbent.

[0011] According to the present invention, even when applied to large-scale production, high-purity medium-molecule pharmaceuticals can be purified with a high recovery rate.

[0012] FIG. 1 shows an enlarged portion of an example chromatogram of the liquid to be treated obtained in the primary treatment. FIG. 2 shows an enlarged portion of an example chromatogram of the liquid to be treated obtained in the primary treatment. FIG. 3 shows an example of the configuration of a primary treatment device. FIG. 4 shows an example of the configuration of a simulated moving bed device. FIG. 5 shows an example of the operation flow of the simulated moving bed device of FIG. 4. FIG. 6 shows the concentration distribution of P component and R component in the packed section of the simulated moving bed device of FIG. 4. FIG. 7 shows the flow directions of the primary treatment liquid and eluent in the packed section when steps 101 and 102 of FIG. 5 are repeated four times.

[0013] The meanings of the terms are as follows. In this specification, a component that has a stronger interaction with the separating agent may be referred to as a P component. A component that has a weaker interaction with the separating agent may be referred to as an R component. When the liquid to be treated is passed through the separating agent, the R component passes through at a faster rate than the P component. Therefore, the R component tends to move ahead in the direction of liquid passage, and the P component tends to remain behind. As a result, the P component and the R component are separated. A liquid that remains after separation of the components and is rich in either the P component or the R component may be referred to as a "separated liquid."

[0014] In this specification, the terms α component and β component are not single impurity components, but are collective terms for components that have weaker and stronger interactions with the separating agent than the medium-molecular-weight drug. The strength of the interaction between the α component and the separating agent varies depending on the separating agent, and also on the composition of the liquid to be treated with the medium-molecular-weight drug.

[0015] In this specification, the use of "to" to indicate a range of values ​​means that the values ​​before and after it are included as the lower and upper limits.

[0016] [Medium-sized molecule drugs] Medium-sized molecule drugs are drugs that mainly contain compounds with a molecular weight of 500 to 10,000. Examples of medium-sized molecule drugs include peptide drugs and nucleic acid drugs.

[0017] Peptide drugs have a molecular weight of 500 to 6000 and mainly contain peptides with multiple amino acid residues. Peptide drugs are broadly divided into conventional peptide drugs composed of natural amino acids and next-generation peptide drugs composed of non-natural amino acids. Conventional peptide drugs generally have 8 to 50 amino acid residues and may have a molecular weight of 500 to 6000. Next-generation peptide drugs generally have 8 to 20 amino acid residues and may have a molecular weight of 500 to 3000.

[0018] Examples of next-generation peptide pharmaceuticals include glucagon-like peptide 1 (GLP-1 receptor agonists) such as dulaglutide, semaglutide, liraglutide, and tirzepatide, as well as leuprorelin, teriparatide, and icatibant.

[0019] Nucleic acid drugs are a general term for pharmaceuticals whose basic structure is nucleotides, which are components of DNA and RNA that control the genetic information of living organisms, and their derivatives. Examples include antisense, siRNA (small interfering RNA), miRNA (microRNA), decoy, aptamer, and CpG oligo (CpG-ODN).

[0020] When separating and purifying medium-sized molecular weight drugs such as peptide drugs and nucleic acid drugs, the interaction between impurities and the separating agent is similar to the interaction between the target medium-sized molecular weight drug and the separating agent, making separation difficult. In the following embodiment, an example will be described in which icatibant, a type of peptide drug, is purified as the target medium-sized molecular weight drug.

[0021] [Outline of Icatibant Separation and Purification] In the separation and purification of icatibant, impurities are separated and removed from a treatment target liquid containing icatibant and multiple impurities. The treatment target liquid is a liquid to be separated in order to recover icatibant. As the treatment target liquid, crude icatibant obtained by solid-phase peptide synthesis is usually used, but is not particularly limited. Crude icatibant contains various impurities in addition to icatibant. For example, a synthetic product from Hamari Pharmaceutical Co., Ltd. can be used.

[0022] The present inventors have come up with the idea of ​​incorporating simulated moving bed chromatography (SMB) into a separation process to increase the recovery rate of icatibant while maintaining its high purity. As a result, they have found that, in order to increase the recovery rate, it is effective to remove, as a pretreatment for applying the SMB, the impurity that has the smallest overlap of peak areas with icatibant in the chromatogram among multiple impurities in the solution to be treated.

[0023] Here, the impurity with the smallest peak area overlap with icatibant is defined as the α component, and the process for selecting the α component will be described in more detail. In the partially enlarged chromatogram of the liquid to be treated shown in Figure 1, peak C1 is the detected peak for icatibant. While impurity peaks C2 and C3 are shown in Figure 1, typical crude icatibant contains many other impurities (not shown) in addition to the impurities corresponding to these two peaks. In this chromatogram, the area of ​​overlap S3 with the icatibant peak area is the smallest, so the component corresponding to peak C3 is selected as the α component. Meanwhile, the area of ​​overlap S2 with the icatibant peak area is larger than the area of ​​overlap S3. Therefore, the component corresponding to peak C2 is not selected as the component to be removed in the pretreatment (primary treatment) process.

[0024] In the present invention, the simulated moving bed process in the secondary treatment step can separate components other than the α component (for example, the β component corresponding to peak C2 in the example shown in Figure 1). In the primary treatment step, in order to maximize the recovery rate of icatibant, the overlapping portion of icatibant and the β component is recovered as a purified liquid. In the secondary treatment step (SMB), impurities such as the β component can be removed, so the content of the β component in the primary treatment liquid may be high. In this way, by using the primary treatment step and the secondary treatment step in combination, it is possible to achieve both a high recovery rate and purity of icatibant.

[0025] In the present invention, α-components are removed in the primary treatment, which is liquid chromatography. Among the multiple impurities in the liquid to be treated, α-components have the smallest overlap with the peak area of ​​icatibant in the chromatogram obtained by the primary treatment step. Therefore, even if α-components are removed in the pretreatment step, the amount of icatibant removed and loss are relatively small. The present inventors discovered that by further performing a simulated moving bed method using icatibant obtained from the primary treatment liquid, a high recovery rate can be achieved while maintaining high purity of icatibant, leading to the completion of the present invention.

[0026] In liquid chromatography in the primary treatment, multiple components can be separated sequentially by utilizing the difference in the interaction of each component in the treatment liquid with the separating agent. For example, as shown in Figure 1, in the case of a treatment liquid containing two types of impurities, an α component corresponding to peak C3 and a β component corresponding to peak C2, the β component, which has a weaker interaction with the separating agent, has a shorter retention time and is therefore separated first. The α component, which has a stronger interaction with the separating agent, has a longer retention time and is therefore separated last.

[0027] However, the liquid to be treated is not limited to containing two types of impurities, and may contain three or more types of impurities. In this case, the components that overlap the peak area of ​​icatibant in the chromatogram of the liquid to be treated are removed in order.

[0028] Several embodiments will be described in detail below with reference to the drawings as appropriate, but the embodiments of the present invention are not limited to the following description. The dimensional ratios in the drawings are set for the convenience of explaining the embodiments and may differ from the actual dimensions.

[0029] [Method for separating and purifying icatibant] The method for separating and purifying icatibant according to one embodiment includes a primary treatment step and a secondary treatment step. Hereinafter, the primary treatment step and the secondary treatment step will be described in order.

[0030] (Primary Treatment Step) In the primary treatment step, a treatment target liquid containing icatibant and multiple impurities is supplied to a column packed with separating agent X to obtain a primary treatment liquid from which the α component has been removed. As already mentioned, among the multiple impurities in the treatment target liquid, the α component is the impurity that has the smallest overlap with the peak area of ​​icatibant in the chromatogram of the treatment target liquid obtained by primary purification. The conditions for removing the α component from the treatment target liquid can be determined by those skilled in the art by changing various conditions such as the type and concentration of the eluent, gradient conditions, injection pressure, flow rate, and temperature.

[0031] When using the primary treatment device 1 shown in Figure 3, the liquid to be treated containing icatibant and multiple impurities is supplied from a pipe 51 to a column 52 packed with a separating agent X. The α component is removed from the liquid to be treated by adjusting the interaction with the separating agent X in the column 52. An eluent is used to adjust the interaction with the separating agent X. Thereafter, the primary treatment liquid from which the α component has been removed is recovered from the liquid to be treated via a pipe 53.

[0032] In the chromatogram of the primary treatment step shown in Figure 2, the fraction of the treatment solution from the start of icatibant peak C1 can be retained to obtain the primary treatment solution, and the β component that flows out before that can be removed. Alternatively, the fraction of the treatment solution corresponding to the bottom of the peak formed by icatibant peak C1 and α component peak C3 (shown in Figure 2) can be retained to obtain the primary treatment solution, and the α component that flows out after that can be removed. As described above, it is preferable to obtain the fraction of the treatment solution from the start of icatibant peak C1 to the bottom of the peak formed by icatibant peak C1 and α component peak C3 as the primary treatment solution (the gray area in Figure 2).

[0033] The α component content of the primary treatment liquid obtained in the primary treatment step is preferably as close to 0 as possible, from the viewpoint of the purity of icatibant in the secondary treatment. For example, 1.0 mass% or less is preferable, 0.5 mass% or less is more preferable, and 0.1 mass% or less is even more preferable. The α component content is obtained as follows: (α component content) = (mass of α component) / ((mass of total impurities) + (mass of icatibant)).

[0034] The purity of icatibant in the primary treatment solution obtained in the primary treatment step is preferably 90% by mass or more, more preferably 94% by mass or more, and even more preferably 97% by mass or more, since this tends to increase the recovery rate of icatibant. The purity of icatibant is obtained as follows: (Icatibant purity) = (Icatibant mass) / ((mass of all impurities) + (Icatibant mass)).

[0035] (Separating agent X in the primary treatment step) Various separating agents used in the field of liquid chromatography can be used without limitation as the separating agent X. Examples include polymeric separating agents made of polymeric materials and inorganic separating agents made of inorganic materials.

[0036] Examples of polymeric materials for the polymeric separating agent include vinyl synthetic polymers, diene synthetic polymers, condensation synthetic polymers, and curable synthetic polymers.

[0037] Examples of vinyl-based synthetic polymers include styrene-based synthetic polymers such as polystyrene (PS), (meth)acrylic-based synthetic polymers such as polymethyl methacrylate (PMMA), acetal-based synthetic polymers such as polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), polyethylene (PE), polypropylene (PP), polyacrylonitrile (PAN), polyvinyl alcohol (PVA), and polyvinyl butyral, polyvinyl acetate (PVAc), polyacrylamide (PAA), and polyvinyl ether-based synthetic polymers.

[0038] Examples of the diene-based synthetic polymer include polybutadiene (PBd) and polyisoprene (PIP).

[0039] Examples of condensation synthetic polymers include polyamide synthetic polymers such as nylon 6 and nylon 66, polyester synthetic polymers such as polyethylene terephthalate (PET) and polylactone, and polyether synthetic polymers such as polycarbonate (PC) and polyoxymethylene (POM).

[0040] Examples of the curable synthetic polymer include polyurethane resin, alkyd resin, phenolic resin, polyaniline (PANI), epoxy-based synthetic polymer, and silicone-based synthetic polymer.

[0041] Other examples of polymeric materials include xylene-based synthetic polymers, furan-based synthetic polymers, terpene-based synthetic polymers, petroleum-based synthetic polymers, ketone-based synthetic polymers, and sulfur-based synthetic polymers such as polycyclic thioethers.

[0042] Examples of inorganic materials for the inorganic separating agent include activated carbon, silica gel, diatomaceous earth, hydroxyapatite, alumina, titanium oxide, magnesia, and polysiloxane.

[0043] Among these, polymeric separating agents are preferred as the separating agent X in terms of separation ability, and synthetic adsorbents are more preferred. Synthetic adsorbents are polymers having a porous structure and a cross-linked structure. Synthetic adsorbents can remove organic matter from the liquid to be treated by adsorption, utilizing the hydrophobic interaction of their hydrophobic moieties.

[0044] The shape of the synthetic adsorbent is not particularly limited, and may take various shapes such as spheres, ellipsoids, cylinders, prisms, etc. Of these, spherical synthetic adsorbents are preferred.

[0045] The average particle size of the synthetic adsorbent is not particularly limited, but may be, for example, in the range of 5 to 100 μm, 8 to 50 μm, or 10 to 30 μm. The average particle size of the synthetic adsorbent can be measured by a known method. For example, it can be obtained by measuring the particle sizes of 100 or more particles using an optical microscope and calculating the volume median diameter from the distribution.

[0046] Among these, the synthetic adsorbents used as the separating agent X are preferably styrene-divinylbenzene synthetic adsorbents, Br-modified styrene-divinylbenzene synthetic adsorbents, and (meth)acrylic synthetic adsorbents, and more preferably styrene-divinylbenzene synthetic adsorbents.

[0047] (Eluant in the primary treatment step) The eluant used in the primary treatment is not particularly limited and may be appropriately selected depending on the state of the liquid to be treated and the treatment conditions. For example, organic solvents include acetonitrile, ethanol, and methanol. Of these, acetonitrile is more preferred. As a buffer for adjusting pH, phosphate, acetate, and ammonium salt are preferred, and phosphate is more preferred.

[0048] (Secondary Treatment Step) In the secondary treatment step, a simulated moving bed method is performed to continuously separate icatibant by supplying the primary treatment liquid to multiple packed sections filled with separating agent Y. Below, an example of performing the secondary treatment step using the simulated moving bed apparatus 2 shown in Figure 4 will be described, but the secondary treatment step is not limited to this example.

[0049] (Separating agent Y in the secondary treatment step) The separating agent used as separating agent Y may be selected from the same separating agent used in the primary treatment. Among them, synthetic adsorbents are preferred. As the type of synthetic adsorbent, styrene-divinylbenzene synthetic adsorbents, Br-modified styrene-di-vinylbenzene synthetic adsorbents, and (meth)acrylic synthetic adsorbents are preferred, and styrene-divinylbenzene synthetic adsorbents are more preferred.

[0050] The average particle size of the separating agent Y is not particularly limited, but may be, for example, in the range of 5 to 100 μm, 10 to 50 μm, or 10 to 30 μm. The average particle size of the synthetic adsorbent can be measured by known methods. For example, it can be obtained by measuring the particle sizes of 100 or more particles using an optical microscope and calculating the volume median diameter from the distribution. The type and average particle size of the separating agent Y may be the same as those of the separating agent X, or may be selected independently.

[0051] (Eluant in the secondary treatment step) The eluant may be the same as the eluant used in the primary treatment, or may be selected independently. The buffer may also be the same as the eluant used in the primary treatment, or may be selected independently.

[0052] Figure 5 is a flowchart showing an example of the operation of the simulated moving bed apparatus 2 in the secondary treatment step. As shown in Figure 5, the simulated moving bed process in the secondary treatment step includes a supply / withdrawal step and a circulation step. In the supply / withdrawal step, a primary treatment liquid and an eluent are supplied to at least two or more different packed sections among a plurality of packed sections (11, 12, 13, 14), respectively, and a separated liquid is withdrawn from each packed section to which the primary treatment liquid and the eluent have been supplied. In the circulation step, the primary treatment liquid and the eluent are not additionally supplied to the plurality of packed sections (11, 12, 13, 14), but are circulated between the plurality of packed sections (11, 12, 13, 14) in the same flow direction as in the supply / withdrawal step.

[0053] 6(a) and 6(b) are diagrams showing the concentration distributions of the P and R components in the packing sections 11 to 14 during the supply / withdrawal step and the circulation step. The horizontal direction indicates the position within the packing sections 11 to 14. In FIG. 6, the left side indicates the upper (more upstream) position within the packing sections 11 to 14. The right side indicates the lower (more downstream) position within the packing sections 11 to 14.

[0054] In Figure 6, the vertical direction represents the concentrations of the P component and the R component at each position. The right arrow represents the flow direction of the primary treatment liquid and the eluent within the packing sections 11 to 14. In this case, the right arrow indicates that the primary treatment liquid and the eluent flow downward within the packing sections 11 to 14. If no right or left arrow is shown, it means that no flow occurs within the packing sections 11 to 14. In Figure 6, the down arrow and the up arrow indicate the points where the primary treatment liquid and the eluent are supplied, and the points where the P fraction, which is a separation liquid rich in the P component, and the R fraction, which is a separation liquid rich in the R component, are extracted. In the figure, the primary treatment liquid is represented by "F," the eluent is represented by "W," the P component and the P fraction are represented by "P," and the R component and the R fraction are represented by "R."

[0055] In Fig. 6, the packing section 13 is designated as the adsorption zone (Zone 1), the packing section 14 is designated as the purification zone (Zone 2), the packing section 11 is designated as the desorption zone (Zone 3), and the packing section 12 is designated as the concentration zone (Zone 4). Separation operation can be carried out continuously by shifting these zones one by one.

[0056] When the primary treatment liquid is passed through the separating agent Y in the packed sections 11 to 14, the R component passes through at a higher rate than the P component, as described above. Therefore, for example, as shown in FIG. 6( a), the R component tends to advance in the direction of liquid passage, while the P component tends to remain behind. In other words, the P component and the R component are separated in the packed sections 11 to 14. Here, if the α component (corresponding to peak C3 in FIG. 1) is a component that is more strongly retained than icatibant in the primary treatment step, the P component (corresponding to peak C1 in FIG. 1) will be enriched in icatibant.

[0057] In the state shown in Figure 6(a), the primary effluent and the eluent are supplied downward from separate supply ports. Simultaneously, a P-component-rich separated liquid and an R-component-rich separated liquid are withdrawn from separate withdrawal ports (step 101: supply and withdrawal step). In this case, the primary effluent on-off valve F3, the eluent on-off valve W1, the connecting line on-off valves X1 and X2, the P-component on-off valve P1, and the R-component on-off valve R3 are open, while the other on-off valves are closed. In this state, the primary effluent can be supplied from the supply port 23 to the packing port 13, and the eluent can be supplied from the supply port 21 to the packing port 11. Furthermore, the P fraction, which is a P-component-rich separated liquid, can be withdrawn from the withdrawal port 31, and the R fraction, which is a R-component-rich separated liquid, can be withdrawn from the withdrawal port 33.

[0058] In this supply / withdrawal step, the P component is eluted using an eluent supplied from the supply unit 21 to the packing unit 11. A portion of the supplied eluent is withdrawn into the pipe HP1 as a P fraction, which is a separation liquid rich in the P component. The remaining eluent that is not withdrawn from the withdrawal unit 31 flows into the packing unit 12 from the pipe HX1. As a result, the eluent moves downward and flows through the packing unit 12 and the packing unit 13. As separation of the P component and the R component progresses in the packing unit 12 and the packing unit 13, the concentration distribution of the P component and the R component also shifts downstream. A primary treatment liquid is then supplied to the packing unit 13, and the R fraction, which is a separation liquid rich in the R component, is withdrawn into the pipe HR3 from the withdrawal unit 33 of the packing unit 13. Here, the amount of liquid withdrawn into the pipe HP1 is a portion of the amount of liquid supplied from the supply unit 21. Therefore, in order to control this flow rate, it is useful to attach a pump to the end of the pipe HP1 to extract the water at a constant flow rate and to adjust the amount extracted with an integrating flow meter.

[0059] In the supply and withdrawal step exemplified here, the following operations may be separately performed: - An operation of supplying the primary treatment liquid from the supply unit 23 and withdrawing the R fraction from the withdrawal unit 33 - An operation of supplying the eluent from the supply unit 21 and withdrawing the P fraction from the withdrawal unit 31 - An operation of supplying the eluent from the supply unit 21 and withdrawing the R fraction from the withdrawal unit 33

[0060] At the end of step 101, the concentration distribution of the P component and the R component will be as shown in Fig. 6(b) In the state shown in Fig. 6(b), the primary treatment liquid and the eluent in the packing section 10 are circulated in a downward flow between the packing sections 10 to proceed with the separation of the multiple components (step 102: circulation step).

[0061] In step 102, the primary treatment liquid and the eluent are not supplied. The connecting line on-off valves X1, X2, and X3 are opened, and the other on-off valves are closed. The pump PM is then operated to circulate the primary treatment liquid and the eluent in the charging sections 10 in a downward flow between the charging sections 10. In this case, by opening the connecting line on-off valves X1, X2, and X3, all of the charging sections 10 are connected via the pipes HX1, HX2, HX3, and HX4 and the bypass line HB. As a result, a circulation path that satisfies the following conditions 1 and 2 is formed in the simulated moving bed apparatus 2. Condition 1: The primary treatment liquid and the eluent are not additionally supplied to the multiple charging sections 10. Condition 2: The primary treatment liquid and the eluent are circulated between the multiple charging sections 10 in a direction from the supply section 20 toward the withdrawal section 30.

[0062] By operating the pump PM, the primary treatment liquid and the eluent can be moved through the circulation path. Performing one circulation step moves the primary treatment liquid and the eluent in the packing section 10 downward by one packing section 10. At this time, separation of the P component and the R component also progresses. As a result, the concentration distribution shifts one packing section 10 to the right in FIG. 6 from the state shown in FIG. 6( a). In other words, the concentration distribution is reproduced with one packing section 10 shifted to the right in FIG. 6. This allows the process to return to step 101 and repeat the same separation process, enabling continuous separation by liquid chromatography.

[0063] Next, as shown in FIG. 5 , it is determined whether or not to terminate separation using the simulated moving bed method (step 103). Separation may be terminated, for example, when a predetermined amount of primary treatment liquid has been treated. Separation may also be terminated when the pressure loss exceeds a predetermined magnitude, or when a predetermined separation operation time has elapsed. If separation is to be terminated (Yes in step 103), the separation operation is stopped (step 104). On the other hand, if separation is not to be terminated (No in step 103), the process returns to step 101. Thereafter, steps 101, 102, and 103 are repeated.

[0064] 7(a) to 7(h) are diagrams showing the flow directions of the primary treatment liquid and eluent in the packing sections 11 to 14 when steps 101, 102, and 103 of FIG. 5 are repeated four times (steps 1 to 4). Here, FIGS. 7(a) and 7(b) show step 1, FIGS. 7(c) and 7(d) show step 2, FIGS. 7(e) and 7(f) show step 3, and FIGS. 7(g) and 7(h) show step 4. In FIG. 7, as in FIG. 6, right arrows indicate downward flow. Furthermore, if no right arrow is shown, it means that no flow is occurring in the packing sections 11 to 14. Downward and upward arrows indicate the points where the primary treatment liquid and eluent are supplied and the points where the P fraction and the R fraction are extracted. In this case, the primary treatment liquid is represented by "F", the eluate by "W", the P component or P fraction by "P", and the R component or R fraction by "R".

[0065] The following Table 1 shows the open state of each on-off valve of the switching unit 40 in each step shown in Fig. 7. On-off valves other than those shown here are closed.

[0066]

[0067] Comparing Figures 7(a) and 7(b), Figures 7(c) and 7(d), Figures 7(e) and 7(f), and Figures 7(g) and 7(h), the flow direction, the supply points for the primary treatment solution and eluent, and the extraction points for the P and R fractions are shifted by one valve to the right (downstream) in the figures for each packing section 10. Also, referring to Table 1, the positions of the on-off valves to be opened are also shifted by one valve to the right (downstream) in the figures for each packing section 10. After steps 1 to 4 are performed, the system returns to the state of step 1. In other words, after Figure 7(h), the system returns to the state of Figure 7(a).

[0068] In the secondary treatment process, the space velocity (SV) is 0.5 h －１ ~10 hours －１ It is preferable to supply the primary treatment liquid to at least one of the multiple packed sections (11, 12, 13, 14) under the conditions. The closer the spatial velocity is to the lower limit of the above-mentioned range, the more likely it is that the purity and recovery rate of icatibant will improve. The closer the spatial velocity is to the upper limit of the above-mentioned range, the higher the productivity will be. The spatial velocity is preferably set to a flow rate that applies a pressure of about 0.8 times the column pressure resistance.

[0069] In the secondary treatment step, the pH of the eluent in the plurality of packing sections (11, 12, 13, 14) is preferably 7.0 or less, more preferably 5.0 or less, and even more preferably 3.0 or less.

[0070] (Mechanism of Action) The icatibant separation and purification method described above removes the α component from the liquid to be treated in the primary treatment step. The α component is the impurity that has the smallest overlap with the peak area of ​​icatibant in the chromatogram of the liquid to be treated. This reduces the loss of icatibant and improves the recovery rate. Then, based on the characteristics of icatibant and the α component, the separating agent X, eluent, separation conditions, etc. are determined. In addition, according to the icatibant separation and purification method, in the secondary treatment step, icatibant can be continuously separated from the primary treatment liquid obtained in the primary treatment step. This improves the recovery rate while maintaining the high purity of icatibant.

[0071] (α component selection step) In one example, the method for purifying icatibant may further include the following α component selection step before the primary treatment step. The α component selection step includes the following steps: a step of determining a first peak (C1) corresponding to icatibant in a chromatogram obtained when the liquid to be treated is supplied to a column packed with separating agent X; a step of determining a second peak (C2) corresponding to an impurity that passed through separating agent X before icatibant, among multiple impurities, in the chromatogram; and a step of determining a third peak (C3) corresponding to an impurity that passed through separating agent X after icatibant. A step of selecting, as the α component, the impurity corresponding to the peak that overlaps the smallest area with the first peak (C1) from the second peak (C2) and the third peak (C3).

[0072] [Post-treatment] After the secondary treatment, post-treatment may be carried out as needed. Examples of post-treatment include solvent removal treatment, cation exchange treatment, and anion exchange treatment.

[0073] [Icatibant Separation and Purification System] The Icatibant separation and purification system according to one embodiment includes a primary treatment device and a simulated moving bed apparatus. The primary treatment device and the simulated moving bed apparatus will be described below in this order.

[0074] (Primary treatment device) The primary treatment device supplies the liquid to be treated, which contains icatibant and multiple impurities, to a column filled with separating agent X, thereby obtaining a primary treatment liquid from which the α component has been removed from the liquid to be treated.

[0075] For example, the primary treatment device 1 shown in Figure 3 has a pipe 51, a column 52, and a pipe 53. According to the primary treatment device 1, the liquid to be treated containing icatibant and multiple impurities can be supplied from the pipe 51 to a column 52 packed with a separating agent X. In the column 52, the interaction with the separating agent X is adjusted to remove the α component from the liquid to be treated, i.e., the impurity that has the smallest overlap with the peak area of ​​icatibant in the chromatogram of the liquid to be treated.

[0076] An eluent is used to adjust the interaction with the separating agent X in the column 52, thereby removing the α component from the liquid to be treated. The eluent used in the primary treatment can be selected appropriately depending on the state of the liquid to be treated and the treatment conditions. Examples include acetonitrile, ethanol, and methanol. Of these, acetonitrile is more preferable. The primary treatment liquid from which the α component has been removed can be recovered and obtained from the pipe 53.

[0077] From the viewpoint of the purity of icatibant in the secondary treatment, the content of the α component in the primary treatment solution obtained in the primary treatment device is preferably as close to 0 as possible. For example, the content is preferably 1.0% by mass or less, more preferably 0.5% by mass or less, and even more preferably 0.1% by mass or less.

[0078] The content of icatibant in the primary treatment solution obtained by the primary treatment device is preferably 90% by mass or more, more preferably 94% by mass or more, and even more preferably 97% by mass or more, because the recovery rate of icatibant by secondary treatment is likely to be high. Since impurities other than the α component can be removed by secondary treatment, icatibant may be contained in the primary treatment solution.

[0079] The details and preferred aspects of the separating agent X are the same as those described for the separation and purification method, and a synthetic adsorbent is preferred. The details and preferred aspects of the synthetic adsorbent are also the same as those described for the separation and purification system.

[0080] (Simulated Moving Bed Apparatus) The simulated moving bed apparatus performs a simulated moving bed process to continuously separate icatibant by supplying the primary treated liquid obtained in the primary treatment apparatus to a plurality of packed sections filled with separating agent Y. The simulated moving bed apparatus is not particularly limited as long as it is an apparatus for performing a simulated moving bed process using the primary treated liquid obtained in the primary treatment apparatus as the liquid to be separated.

[0081] For example, the simulated moving bed apparatus 2 shown in Figure 4 can perform a simulated moving bed process for continuously separating icatibant by supplying the primary treatment liquid to multiple packed sections 10 (11, 12, 13, 14) filled with separating agent Y. The simulated moving bed apparatus 2 has the packed sections 10, a supply section 20, a withdrawal section 30, and a switching section 40.

[0082] The packed section 10 is a section for separating icatibant from impurities. Four packed sections 10 are provided in one simulated moving bed apparatus 2. In the example shown in FIG. 4, packed sections 11, 12, 13, and 14 (packed sections 11 to 14) are illustrated as the packed sections 10. Hereinafter, when there is no need to distinguish between the packed sections 11, 12, 13, and 14, they may be simply referred to as packed sections 10. When the separation conditions need to be adjusted based on the state of the primary treatment liquid, etc., five or more packed sections 10 may be provided in one simulated moving bed apparatus 2.

[0083] The filling section 10 is filled with a separating agent Y for separating icatibant contained in the primary treatment liquid. Examples of separating agent Y include the same separating agents as those exemplified for separating agent X in the primary treatment device, and synthetic adsorbents are preferred. In the entire icatibant separation and purification system, separating agent Y may be the same as or different from separating agent X.

[0084] The separation column packed with separating agent Y is preferably a packed column having no empty column at the top. The packed section 10 may be, for example, a column. The packed section 10 has a space inside for packing with separating agent Y. Examples of materials for the packed section 10 include steel plate and resin. The liquid-contacting part may be lined with rubber, but this is not particularly limited. The shape of the packed section 10 is not particularly limited, but an example is a substantially cylindrical column.

[0085] The supply section 20 is a section for separately supplying the primary treatment liquid and the eluent to each of the multiple packing sections. The eluent is a liquid used to recover a separation liquid rich in icatibant. The eluent is used to develop the components adsorbed in each packing section. The strength of the interaction between the separating agent Y and the adsorbed components can be adjusted by the eluent. By adjusting the interaction between the separating agent Y and the components by the concentration of the eluent, each component can be separated and eluted.

[0086] The eluent used in the secondary treatment can be selected appropriately depending on the state of the primary treatment liquid and the treatment conditions. Examples include acetonitrile, methanol, and ethanol. Among these, acetonitrile is preferred. The eluent used in the secondary treatment may be the same as or different from the eluent used in the primary treatment. From the viewpoint of ease of management, it is preferable that the eluent used in the secondary treatment is the same as the eluent used in the primary treatment.

[0087] 4, the supply unit 20 is formed as a supply port at the top of each filling unit. More specifically, supply units 21, 22, 23, and 24 (supply units 21 to 24) are shown as the supply unit 20. Hereinafter, when there is no need to distinguish between the supply units 21, 22, 23, and 24, they may be simply referred to as supply unit 20.

[0088] The withdrawal section 30 is a section for withdrawing the separated liquid from each of the multiple packed sections. In the example shown in Figure 4, the withdrawal section 30 is formed as a discharge port at the bottom of each packed section. To explain in more detail, withdrawal sections 31, 32, 33, and 34 (withdrawal sections 31 to 34) are shown as the withdrawal section 30. Hereinafter, when there is no need to distinguish between the withdrawal sections 31, 32, 33, and 34, they may be simply referred to as withdrawal section 30.

[0089] The switching unit 40 is a component for switching the flow paths. In the example shown in FIG. 4 , the switching unit 40 includes a plurality of on-off valves. By opening and closing the plurality of on-off valves, the flow paths of the primary treatment liquid, eluent, and separation liquid can be switched. More specifically, the switching unit 40 includes eluent on-off valves W1, W2, W3, and W4, primary treatment liquid on-off valves F1, F2, F3, and F4, connection path on-off valves X1, X2, X3, and X4, R component on-off valves R1, R2, R3, and R4, and P component on-off valves P1, P2, P3, and P4.

[0090] The simulated moving bed apparatus 2 has a pipe HW for supplying an eluent, a pipe HW1 for supplying the eluent from the pipe HW to the filling section 11, a pipe HW2 for supplying the eluent from the pipe HW to the filling section 12, a pipe HW3 for supplying the eluent from the pipe HW to the filling section 13, and a pipe HW4 for supplying the eluent from the pipe HW to the filling section 14. Eluent on-off valves W1 to W4 are provided in the pipes HW1 to HW4, respectively, and control the supply of the eluent to the filling sections 11 to 14.

[0091] The simulated moving bed apparatus 2 has a pipe HF for supplying the primary treated liquid, a pipe HF1 for supplying the primary treated liquid from the pipe HF to the filling section 11, a pipe HF2 for supplying the primary treated liquid from the pipe HF to the filling section 12, a pipe HF3 for supplying the primary treated liquid from the pipe HF to the filling section 13, and a pipe HF4 for supplying the primary treated liquid from the pipe HF to the filling section 14. Primary treated liquid on-off valves F1 to F4 are provided in the pipes HF1 to HF4, respectively, and control the supply of the primary treated liquid to the filling sections 11 to 14.

[0092] The simulated moving bed apparatus 2 has, as connection paths connecting each of the filling sections 10, a pipe HX1 connecting the withdrawal section 31 of the filling section 11 with the supply section 22 of the filling section 12, a pipe HX2 connecting the withdrawal section 32 of the filling section 12 with the supply section 23 of the filling section 13, a pipe HX3 connecting the withdrawal section 33 of the filling section 13 with the supply section 24 of the filling section 14, and a pipe HX4 connecting the withdrawal section 34 of the filling section 14 with the supply section 21 of the filling section 11. Connection path on-off valves X1 to X4 are provided in the pipes HX1 to HX4, respectively, to control the flow of liquid between the filling sections 11 to 14.

[0093] A bypass path HB is provided at the location of the connection path on-off valve X4 of the pipe HX4. A pump PM is provided in the bypass path HB. Although the bypass path HB and the pump PM are installed in the pipe HX4, they may be installed in any of the pipes HX1 to HX4, or may be installed in multiple positions (for example, all positions) of the pipes HX1 to HX4.

[0094] The simulated moving bed apparatus 2 has a pipe HR for withdrawing the R fraction, a pipe HR1 for withdrawing the R fraction from the packed section 11 to the pipe HR, a pipe HR2 for withdrawing the R fraction from the packed section 12 to the pipe HR, a pipe HR3 for withdrawing the R fraction from the packed section 13 to the pipe HR, and a pipe HR4 for withdrawing the R fraction from the packed section 14 to the pipe HR. R component on-off valves R1 to R4 are provided on the pipes HR1 to HR4, respectively, and control the withdrawal of the separated liquid from the packed sections 11 to 14.

[0095] The simulated moving bed apparatus 2 has a pipe HP for withdrawing the P fraction, a pipe HP1 for withdrawing the P fraction from the packed section 11 to the pipe HP, a pipe HP2 for withdrawing the P fraction from the packed section 12 to the pipe HP, a pipe HP3 for withdrawing the P fraction from the packed section 13 to the pipe HP, and a pipe HP4 for withdrawing the P fraction from the packed section 14 to the pipe HP. P component on-off valves P1 to P4 are provided on the pipes HP1 to HP4, respectively, and control the withdrawal of the separated liquid from the packed sections 11 to 14.

[0096] In order to maintain a constant solubility of the solute, it is preferable to maintain a constant temperature inside the simulated moving bed apparatus 2. To maintain a constant temperature, the piping and columns may be covered with heat insulating materials, or a heat exchanger may be installed in a location where the temperature is likely to fluctuate. Ultrasound may also be applied to promote the dissolution of the solute components.

[0097] (Mechanism of Action) The icatibant separation and purification system described above removes α-components from the liquid to be treated using a primary treatment device. The α-component is the impurity that has the smallest overlap with the peak area of ​​icatibant in the chromatogram of the liquid to be treated. This reduces the loss of icatibant and improves the recovery rate. The separating agent X, eluent, and separation conditions are then determined based on the characteristics of icatibant and the α-component. Additionally, the icatibant separation and purification system allows icatibant to be continuously separated from the primary treatment liquid obtained in the primary treatment device using a simulated moving bed device. This improves the recovery rate while maintaining the high purity of icatibant.

[0098] Although the embodiments of the present invention have been described above based on specific examples, each embodiment is presented as an example and does not limit the scope of the present invention. Each embodiment described in this specification can be modified in various ways without departing from the spirit of the invention, and can be combined with features described in other embodiments to the extent possible.

[0099] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following description.

[0100] Example 1 (α component selection step) Crude icatibant synthesized by Hamari Pharmaceutical Co., Ltd. was obtained and prepared. Liquid chromatography was performed on the synthetic adsorbent, eluent, and crude icatibant under the following conditions. Purification device: "Medium-high pressure preparative purification device Purif-Rp2" manufactured by SHOKO SCIENTIFIC. Separation agent X: Styrene divinylbenzene ("MCIGEL CHP20 / P10" manufactured by Mitsubishi Chemical Corporation, average particle size 10 μm). Column size: 20φ×15 mmh (47 mL). Eluent A: Acetonitrile / 50 mM Na ３ P.O. ４ (pH 2.0, H ３ P.O. ４ ) = 5 / 95 Eluent B: acetonitrile / 50 mM Na ３ P.O. ４ (pH 2.0, H ３ P.O. ４ ) / H ２ O=8 / 1 / 1 Flow rate: 18 ml / min Raw material: Crude ikatibant 100 mg / mL Injection amount: 9 mL Temperature: 25°C Detection: UV (220 nm, Prep cell)

[0101] The gradient slopes (one cycle) of eluent A and eluent B are shown in Table 2 below.

[0102]

[0103] The resulting chromatogram is shown in a partially enlarged view in Figure 1. The component corresponding to peak C3, which has the smallest area overlap with the icatibant peak, was selected as the α component.

[0104] (Primary Treatment) Primary treatment was carried out under the same conditions as those for the selection of the α component described above, and the fraction between 12.00 minutes and 14.25 minutes in the cycle shown in Table 2 was collected as the icatibant primary treatment solution.

[0105] After the primary treatment, the purity of icatibant was 97.0% by mass, the content of α component was 0.7% by mass, and the recovery rate of icatibant was 98.0%.

[0106] (Secondary Treatment) A separation test of the primary treatment liquid obtained in the primary treatment was carried out using the simulated moving bed apparatus 2 shown in Figure 4. The secondary treatment conditions were as follows: Purification apparatus: "LaboFine" manufactured by Mitsubishi Chemical Aqua Solutions Co., Ltd. Separation agent Y: styrene-divinylbenzene ("MCIGEL CHP20 / P20" manufactured by Mitsubishi Chemical Co., Ltd., average particle size 20 μm) Column size: 4.6φ x 250 mmh x 4 columns (16.6 mL) Eluent: acetonitrile / 50 mM Na ３ P.O. ４ (pH 2.0, H ３ P.O. ４ )=20 / 80 Flow rate: 0.8ml / mL (Space velocity (SV): 2.9h －１ ) Raw material: refined liquid obtained in the primary treatment process Temperature: 25°C

[0107] Table 3 shows the liquid volume for each step in any one of steps 1 to 4 shown in Figure 7. In addition, Run 1 shown in Table 3 is a condition in which priority is given to the purity of icatibant, and Run 2 is a condition in which priority is given to the recovery rate of icatibant.

[0108]

[0109] In Run 1, where priority was given to the purity of icatibant, the purity of icatibant was 99.3% and the recovery rate was 95.5% or more. In Run 2, where priority was given to the recovery rate of icatibant, the purity of icatibant was 99.2% and the recovery rate was 97.5% or more.

[0110] The total recovery rate of icatibant in the primary and secondary treatments was 93.6% in Run 1 and 95.5% in Run 2. The total recovery rate was calculated using the following formula: (Total recovery rate) = (Recovery rate of icatibant in primary treatment) x (Recovery rate of icatibant in secondary treatment).

[0111] Comparative Example 1 Under the same primary treatment conditions as in Example 1, the fraction from 12.50 minutes to 14.25 minutes in the cycle shown in Table 2 was collected to separate the α component and the β component, and icatibant was purified using only the primary treatment.

[0112] As a result, when icatibant was purified by only the primary treatment, the purity of icatibant was 99.6% and the recovery rate was 82.0%.

[0113] Comparative Example 2 Icatibant was purified by isocratic elution (not the simulated moving bed method) using the primary treatment solution obtained in Example 1. The separation conditions were the same as those in the secondary treatment step in Example 1, except for the raw material supply amount: 0.8 ml.

[0114] As a result, fractionation was performed to obtain icatibant with a purity of 99% or more, resulting in an icatibant purity of 99.6% and a recovery rate of 86.3%. The total recovery rate of icatibant combined with the primary treatment was 84.6%.

[0115] From the results of Example 1, it was confirmed that by combining the primary treatment and the secondary treatment, highly pure icatibant can be purified with a high recovery rate even when applied to large-scale production.

[0116] [Example 2] Crude icatibant was purified under the same conditions as in Example 1, except that Sepabeads SP20SS (average particle size 70 μm) manufactured by Mitsubishi Chemical Corporation was used as separating agent Y in the secondary treatment. The results are shown in Table 4.

[0117] [Example 3] Secondary treatment space velocity (SV) 9.8 h －１ The crude icatibant was purified under the same conditions as in Example 1, except that the temperature was changed to: The results are shown in Table 4.

[0118] [Comparative Example 3] Purification of crude icatibant was attempted under the same conditions as in Example 1, except that the crude icatibant was used directly for the secondary treatment without carrying out the primary treatment in Example 1. However, the impurities could not be peeled from the separating agent, and purification and separation were not possible.

[0119]

[0120]

[0121] According to the present invention, even when applied to large-scale production, high-purity medium-molecule pharmaceuticals can be purified with a high recovery rate.

[0122] 1...primary treatment device, 2...simulated moving bed device, 10 (11, 12, 13, 14)...filling section, 20 (21, 22, 23, 24)...supply section, 30 (31, 32, 33, 34)...withdrawal section, 40...switching section

Claims

1. A method for separating and purifying a medium molecular weight drug, comprising: a primary treatment step of supplying a liquid to be treated, which contains a medium molecular weight drug as a separation target and a plurality of impurities, to a column packed with separating agent X to obtain a primary treatment solution from the liquid to be treated of which the following alpha component has been removed; and a secondary treatment step of performing a simulated moving bed method to continuously separate the medium molecular weight drug by supplying the primary treatment solution to a plurality of packing sections packed with separating agent Y, wherein the medium molecular weight drug is a peptide drug or a nucleic acid drug containing a compound having a molecular weight of 500 to 10,000; the alpha component is the impurity, among the plurality of impurities in the liquid to be treated, which has the smallest overlap with the peak area of ​​the medium molecular weight drug in the chromatogram of the liquid to be treated obtained by the primary treatment step.

2. The separation and purification method according to claim 1, wherein the medium-sized molecule drug is icatibant.

3. The separation and purification method according to claim 1 or 2, further comprising the following α component selection step prior to the primary treatment step, the α component selection step comprising: a step of determining a first peak corresponding to the medium molecular weight drug in a chromatogram obtained when the liquid to be treated is supplied to a column packed with the separating agent X; a step of determining a second peak corresponding to an impurity that has passed through the separating agent X before the medium molecular weight drug, among the plurality of impurities, in the chromatogram; a step of determining a third peak corresponding to an impurity that has passed through the separating agent X after the medium molecular weight drug; and a step of selecting, among the second peak and the third peak, an impurity that corresponds to the peak having the smallest overlapping area with the first peak as the α component.

4. A separation and purification method as described in claim 1 or 2, in which, in the chromatogram of the primary treatment step, a fraction of the treatment target liquid from the starting point of a first peak corresponding to the medium molecular weight drug is retained to obtain a primary treatment liquid, and the impurities that flow out before obtaining the primary treatment liquid are removed.

5. A separation and purification method as described in claim 1 or 2, in which a fraction of the liquid to be treated corresponding to the peak formed by the medium molecular weight drug and the peak of the alpha component in the chromatogram of the primary treatment step is retained to obtain a primary treatment liquid, and the alpha component that flows out after obtaining the primary treatment liquid is removed.

6. The separation and purification method according to claim 1 or 2, wherein the separating agent X is a synthetic adsorbent.

7. The separation and purification method according to claim 1 or 2, wherein the secondary treatment step comprises: a supply / withdrawal step of supplying the primary treatment liquid and the eluent to at least two or more mutually different filling sections among the plurality of filling sections, and withdrawing a separated liquid from each of the filling sections to which the primary treatment liquid and the eluent have been respectively supplied; and a circulation step of circulating the primary treatment liquid and the eluent among the plurality of filling sections in the same flow direction as in the supply / withdrawal step, without additionally supplying the primary treatment liquid and the eluent to the plurality of filling sections.

8. The separation and purification method according to claim 1 or 2, wherein the separating agent Y is a synthetic adsorbent.

9. In the secondary treatment step, the space velocity is 0.5 h －１ ~10h －１ 3. The method for separation and purification according to claim 1, wherein the primary treatment liquid is supplied to at least one of the plurality of packing sections under the condition:

10. A separation and purification method according to claim 1 or 2, wherein in the secondary treatment step, the pH of the eluent is adjusted to 7.0 or less.

11. A system for separating and purifying medium-sized molecular drugs, comprising: a primary treatment device that supplies a treatment target liquid containing a medium-sized molecular drug as a separation target and a plurality of impurities to a column packed with separating agent X to obtain a primary treatment liquid in which at least a portion of the alpha component described below has been removed from the treatment target liquid; and a simulated moving bed apparatus that performs a simulated moving bed method to continuously separate the medium-sized molecular drug by supplying the primary treatment liquid to a plurality of packing sections packed with separating agent Y, wherein the medium-sized molecular drug is a peptide drug or a nucleic acid drug containing a compound having a molecular weight of 500 to 10,000; the alpha component is the impurity that, among the plurality of impurities in the treatment target liquid, has the smallest overlap with the peak area of ​​the icatibant in the chromatogram of the treatment target liquid obtained by the primary treatment device.

12. The separation and purification system according to claim 11, wherein the separating agent X is a synthetic adsorbent.

13. The separation and purification system according to claim 11 or 12, wherein the simulated moving bed apparatus has a supply section for separately supplying the primary treatment liquid and the eluent to each of the multiple packing sections, and a withdrawal section for withdrawing the separated liquid from each of the multiple packing sections, and a circulation path that satisfies the following conditions 1 and 2 is formed in the simulated moving bed apparatus. Condition 1: The primary treatment liquid and the eluent are not additionally supplied to the multiple packing sections. Condition 2: The primary treatment liquid and the eluent are circulated between the multiple packing sections in a direction from the supply section toward the withdrawal section.

14. The separation and purification system according to claim 11 or 12, wherein the separating agent Y is a synthetic adsorbent.

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