RNA helicase inhibitor purification process based on solvent extraction

Abstract

The invention discloses an RNA helicase inhibitor purification process based on solvent extraction, and relates to the technical field of RNA helicase inhibitors, the RNA helicase inhibitor purification process comprises a shell, an extractor, a washer, a first motor, a first delivery pipe, a first discharge pipe, a first rotating rod, a first stirring impeller and a liquid pump; the extractor is fixedly connected to the interior of the shell; the washer is fixedly connected to the right side of the extractor in the shell; an extraction function and a washing function are integrated in the same device through an integrated structural design, and an extracted solvent phase containing a target object can directly enter a washing link in the device without being transferred, so that the problems of microbial pollution, target object activity loss and residue loss caused by a transfer process in a traditional step-by-step process are effectively avoided; the purification yield and the product purity are obviously improved; the operation process is greatly simplified, the production cycle is shortened, the process complexity and the production cost are reduced, and the stability and the practicability of the RNA helicase inhibitor purification process are integrally improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of RNA helicase inhibitors, and particularly relates to a solvent extraction-based RNA helicase inhibitor purification process. BACKGROUND

[0002] As a class of biological active substances with antiviral and antitumor potential, the purification process of RNA helicase inhibitors is a key link for industrial application, and the core goal is to remove protein, nucleic acid, endotoxin and other impurities from biological expression systems or chemical synthesis products to obtain high-purity and high-activity target products, and extraction and washing are the core steps for preliminary separation and targeted removal of impurities in the purification process.

[0003] In the existing purification process of RNA helicase inhibitors, extraction and washing operations are usually completed in steps by independent equipment: first, the crude extract is extracted by devices such as separatory funnels and centrifugal extractors to transfer the target inhibitor to a specific solvent phase and achieve preliminary layering; after layering is completed, the solvent phase containing the target substance is taken out from the extraction equipment manually or by a delivery pump and transferred to a dedicated washing device such as a washing tank or another centrifugal device, and washing liquid is added for mixing and cleaning to remove residual impurities, and then layering separation is performed again.

[0004] The above-mentioned step-by-step processing method has obvious defects. The solvent phase containing the target substance after extraction needs to be exposed to the external environment or contact the delivery pipeline during the transfer process, which is easy to be contaminated by microorganisms or lose the activity of the target substance; at the same time, the transfer steps between independent equipment increase the complexity of process operation, prolong the production cycle, and may also cause the loss of part of the target product during the transfer process, reducing the purification yield. SUMMARY

[0005] The present application aims to provide a solvent extraction-based RNA helicase inhibitor purification process to solve the technical problems in the prior art that the solvent phase containing the target substance after extraction needs to be exposed to the external environment or contact the delivery pipeline during the transfer process, which is easy to be contaminated by microorganisms or lose the activity of the target substance; at the same time, the transfer steps between independent equipment increase the complexity of process operation, prolong the production cycle.

[0006] The technical problems solved by the present application can be achieved by the following technical solutions: The application discloses a solvent extraction-based RNA helicase inhibitor purification process, which comprises a shell, an extractor, a washer, a first motor, a first conveying pipe, a first discharge pipe, a first rotating rod, a first stirring vane and a liquid pump.

[0007] Further, the extractor is internally fixed with a first layered hole plate, and the first rotating rod is rotationally connected with the first layered hole plate.

[0008] Further, the top end of the washer is fixed with a second motor, the output end of the second motor is fixed with a second rotating rod, the second rotating rod is fixed with a second stirring vane, the bottom end of the washer is fixed with a second discharge pipe, the bottom end of the second rotating rod is fixed with uniform blades, and the top end of the washer is fixed with a second conveying pipe.

[0009] Further, the top end of the washer is internally fixed with an air cylinder, the bottom end of the air cylinder is fixed with a second layered hole plate, and the second layered hole plate is slidingly connected in the interior of the washer.

[0010] Further, the right side of the first rotating rod is fixed with a motor, the output end of the motor is fixed with a third rotating rod, the outer side wall of the third rotating rod is fixed with a stirring blade, the middle outer side wall of the third rotating rod is fixed with a swing blade, the top end of the first layered hole plate is fixed with a magnet piece, and the magnet piece is arranged below the swing blade.

[0011] Further, the right side of the first rotating rod is fixed with a wall scraping plate, and the wall scraping plate is rotationally connected in the interior of the extractor.

[0012] Further, the inner side wall of the shell is fixed with a heater, the side end of the heater is fixed with a circulating pipe, the interior of the extractor is provided with a heating cavity, and the bottom end of the circulating pipe is arranged in the heating cavity.

[0013] The solvent extraction-based RNA helicase inhibitor purification process comprises the following steps. S1, pretreat the raw material containing RNA helicase inhibitor, destroy the cell structure by high-pressure homogenization, ultrasonic crushing or enzymatic hydrolysis, or remove solid impurities by dissolving in organic solvent and filtering, then obtain the clear crude extract by centrifugation or membrane filtration; then inject the crude extract into the extractor 2 of the shell 1, add the appropriate extractant in proportion, adjust the pH to the vicinity of the isoelectric point of the inhibitor and optimize the salt concentration, realize the full mixing of the two phases by the multidirectional cooperative stirring in the device, make the target inhibitor transfer from the aqueous phase to the organic phase, and complete the preliminary separation; S2, after the extraction is completed, the organic phase containing the target inhibitor does not need to be transferred, and directly enters the scrubber 3 through the flow guide structure in the device and contacts with the preset scrubbing liquid; by using the design of different stirring intensity and direction of the device, the full contact of the scrubbing liquid and the organic phase is ensured, the residual salts, RNase, endotoxin and other impurities are removed efficiently, the emulsification phenomenon is inhibited, and the two-phase layering is accelerated, the separated impurity-containing scrubbing liquid is discharged through a special outlet, and the organic phase remains in the device for subsequent treatment, so that the seamless connection of extraction and washing is realized, and the pollution and loss caused by intermediate transfer are avoided; S3, the organic phase after washing is transferred to the aqueous phase by back extraction, or is directly sent into the concentration unit, the target product is concentrated and desalted by using methods such as reduced pressure evaporation and ultrafiltration centrifugation, then the trace impurities, polymers and residual solvents are further removed by the refining process combined with affinity chromatography, ion exchange chromatography and gel filtration chromatography, after the purity is verified by SDS-PAGE electrophoresis and HPLC detection, the bacteria removal filtration is carried out, finally the high-purity RNA helicase inhibitor is divided and stored under appropriate conditions, so that the activity and stability of the product are ensured, and the experimental or medicinal requirements are met.

[0014] The beneficial effects of the present application are as follows: the extraction function and the washing function are integrated in the same device through the integrated structure design, the solvent phase containing the target substance after extraction can directly enter the washing link in the device without transfer, the problems of microbial contamination, loss of target substance activity and residual loss caused by transfer process in the traditional step-by-step process are effectively avoided, the purification yield and product purity are significantly improved; the operation process is greatly simplified, the production cycle is shortened, the process complexity and production cost are reduced, and the stability and practicality of the RNA helicase inhibitor purification process are improved as a whole. BRIEF DESCRIPTION OF DRAWINGS

[0015] The present application will be further described below with reference to the drawings.

[0016] Figure 1 is the process flow chart of the present application; Figure 2 is the overall structure perspective view of the present application; Figure 3 is the schematic view of the shell structure in the present application; Figure 4 is the present applicationFigure 3 Enlarged view at A in the middle; Figure 5 is the present application Figure 3 Enlarged view at B in the middle; In the figure: 1, shell; 2, extractor; 3, washer; 4, No. 1 motor; 5, No. 2 motor; 6, No. 1 conveying pipe; 7, No. 1 discharge pipe; 8, No. 1 rotating rod; 9, No. 1 stirring impeller; 10, liquid pump; 11, No. 2 rotating rod; 12, No. 2 stirring impeller; 13, No. 2 discharge pipe; 14, No. 1 layered orifice plate; 15, No. 2 layered orifice plate; 16, air cylinder; 17, uniform blade; 18, heater; 19, circulating pipe; 20, heating cavity; 21, wall scraping plate; 22, motor; 23, No. 3 rotating rod; 24, stirring piece; 25, oscillating blade; 26, magnet piece; 27, No. 2 conveying pipe. DETAILED DESCRIPTION

[0017] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of the present application.

[0018] As Figures 1-5 shown, the RNA helicase inhibitor purification process based on solvent extraction includes a shell 1, an extractor 2, a washer 3, a No. 1 motor 4, a No. 1 conveying pipe 6, a No. 1 discharge pipe 7, a No. 1 rotating rod 8, a No. 1 stirring impeller 9, and a liquid pump 10. The extractor 2 is fixedly connected to the inside of the shell 1. The washer 3 is fixedly connected to the right side of the extractor 2 inside the shell 1. The No. 1 motor 4 is fixedly connected to the top end of the extractor 2. The No. 1 rotating rod 8 is fixedly connected to the output end of the No. 1 motor 4. The No. 1 conveying pipe 6 is fixedly connected to the top end of the extractor 2 and the left side of the No. 1 motor 4. The No. 1 stirring impeller 9 is fixedly connected to the outer side wall of the No. 1 rotating rod 8. The No. 1 discharge pipe 7 is fixedly connected to the bottom end of the extractor 2. The liquid pump 10 is arranged between the extractor 2 and the washer 3.

[0019] In operation, the materials are conveyed to the inside of the extractor 2 through the No. 1 conveying pipe 6. After the materials are conveyed, the No. 1 rotating rod 8 is rotated by the No. 1 motor 4. The rotation of the No. 1 rotating rod 8 causes the No. 1 stirring impeller 9 to rotate, thereby stirring the materials in the extractor 2 for extraction treatment. After the extraction treatment, the required liquid materials are conveyed to the inside of the washer 3 by the liquid pump 10, and the unnecessary liquid materials are discharged through the No. 1 discharge pipe 7. The liquid materials enter the inside of the washer 3 for washing treatment. The materials after the extraction treatment do not need to be transferred and can be directly conveyed to the washer 3 for washing by the liquid pump 10.

[0020] The inside of the extractor 2 is fixed with a first layered hole plate 14, and the first rotating rod 8 is rotationally connected with the first layered hole plate 14.

[0021] The rotation of the first stirring impeller 9 can make the material pass through the first layered hole plate 14, and the inside of the first layered hole plate 14 is provided with a plurality of holes. The first layered hole plate 14 is equivalent to a "fine hole sieve". When the liquid material passes through the small holes of the first layered hole plate 14, it will be forcibly divided into small droplets. The water phase and the organic phase in these small droplets originally have a tendency to separate. After passing through the small holes, the "clustering" force is lost, and they will quickly re-aggregate into two layers. The layering speed can be significantly accelerated, and the material layering effect can be improved through the first layered hole plate 14. The top end of the washing device 3 is fixed with a second motor 5; the output end of the second motor 5 is fixed with a second rotating rod 11; the second rotating rod 11 is fixed with a second stirring impeller 12; the bottom end of the washing device 3 is fixed with a second discharge pipe 13; the bottom end of the second rotating rod 11 is fixed with a uniform blade 17; and the top end of the washing device 3 is fixed with a second conveying pipe 27.

[0022] The material enters the inside of the washing device 3 through the liquid pump 10, and the washing liquid is conveyed into the inside of the washing device 3 through the second conveying pipe 27. The second motor 5 makes the second rotating rod 11 rotate, and the rotation of the second rotating rod 11 makes the second stirring impeller 12 rotate and stir. The material in the washing device 3 is washed and treated. The rotation of the second rotating rod 11 makes the uniform blade 17 rotate to stir the bottom of the washing device 3. Then the material is discharged through the second discharge pipe 13. The inside of the top end of the washing device 3 is fixed with an air cylinder 16; the bottom end of the air cylinder 16 is fixed with a second layered hole plate 15, and the second layered hole plate 15 is slidingly connected in the inside of the washing device 3.

[0023] The inside of the second layered hole plate 15 is provided with a plurality of holes. When the liquid material passes through the small holes of the second layered hole plate 15, it will be forcibly divided into small droplets. The water phase and the organic phase in these small droplets can improve the washing treatment effect through the second layered hole plate 15. The height of the second layered hole plate 15 can be adjusted according to the amount of material through the air cylinder 16. The right side of the first rotating rod 8 is fixed with a motor 22; the output end of the motor 22 is fixed with a third rotating rod 23; the outer side wall of the third rotating rod 23 is fixed with a stirring blade 24; the middle outer side wall of the third rotating rod 23 is fixed with a swing blade 25; the top end of the first layered hole plate 14 is fixed with a magnet piece 26, and the magnet piece 26 is arranged below the swing blade 25.

[0024] The rotation of the first rotating rod 8 causes the first stirring impeller 9 to rotate, stirring the material inside the extractor 2. The rotation of the first rotating rod 8 also causes the motor 22 and the third rotating rod 23 to rotate together. The motor 22 then rotates the third rotating rod 23, which in turn rotates the stirring blade 24. The direction of rotation of the stirring blade 24 is different from that of the first stirring impeller 9, creating different stirring paths and improving the stirring effect on the material. The rotation of the first rotating rod 8 causes the oscillating blade 25 to rotate above the magnet plate 26. An iron block is located at the end of the oscillating blade 25. The magnetic attraction of the magnet plate 26 attracts the oscillating blade 25, causing it to bend and move. The first layered perforated plate 14 is equipped with multiple sets of magnets 26 at different positions, which can change the bending state of the oscillating blade 25 several times when it rotates once on the first layered perforated plate 14. The bending of the oscillating blade 25, combined with the rotation of the third rotating rod 23, can change the stirring path and produce different stirring effects. A single stirring direction is prone to forming liquid dead zones in the corners and central areas of the equipment, resulting in insufficient contact between the raw material liquid and the extractant. The convection formed by different stirring directions will impact each other and carry away the liquid in the dead zone, allowing the two phases to form a three-dimensional circulating mixture in the equipment, greatly reducing the uncontacted area, so that the target inhibitor can be transferred to the solvent phase more quickly and fully, while allowing the washing liquid to react efficiently with residual impurities. A scraper plate 21 is fixedly connected to the right side of the first rotating rod 8, and the scraper plate 21 is rotatably connected inside the extractor 2.

[0025] The rotation of the first rotating rod 8 causes the scraper plate 21 to rotate. The rotation of the scraper plate 21 can scrape the material on the inner wall of the extractor 2, preventing the material from adhering to the inner wall of the extractor 2, and also preventing material waste and the inability to stir the material. A heater 18 is fixedly connected to the inner wall of the housing 1; a circulation pipe 19 is fixedly connected to the side end of the heater 18; a heating chamber 20 is opened inside the extractor 2, and the bottom end of the circulation pipe 19 is located in the heating chamber 20.

[0026] The heater 18 is equipped with a water pump and a heater. It heats the water according to the needs of the extractor 2. Hot water flows into the heating chamber 20 through the circulation pipe 19 and then flows into the heater 18. The water is continuously heated and then enters the heating chamber 20 to heat the extractor 2 as needed. The RNA helicase inhibitor purification process based on solvent extraction includes the following steps: S1, pretreatment of raw materials containing RNA helicase inhibitors, destruction of cell structure by high-pressure homogenization, ultrasonic crushing or enzymatic method, or removal of solid impurities by organic solvent dissolution and filtration, then centrifugation, membrane filtration to obtain clear crude extract; then the crude extract is injected into the extractor 2 of the shell 1, the appropriate extractant is added in proportion, the pH is adjusted to the isoelectric point of the inhibitor and the salt concentration is optimized, the two phases are fully mixed by multidirectional cooperative stirring in the device, the target inhibitor is transferred from the aqueous phase to the organic phase, and the preliminary separation is completed; S2, after extraction, the organic phase containing the target inhibitor does not need to be transferred, and is directly introduced into the scrubber 3 through the guide structure inside the device and contacted with the pre-set scrubbing liquid; by using the design of different stirring intensity and direction of the device, the residual salts, RNase, endotoxin and other impurities in the scrubbing liquid are removed efficiently while ensuring the full contact of the scrubbing liquid with the organic phase, inhibiting the emulsification phenomenon and accelerating the phase separation, the separated impurity-containing scrubbing liquid is discharged through a special outlet, and the organic phase remains in the device for subsequent treatment, realizing seamless connection of extraction and washing, avoiding pollution and loss caused by intermediate transfer; S3, the organic phase after washing is transferred to the aqueous phase by back extraction, or is directly sent to the concentration unit, and the target product is concentrated and desalted by vacuum evaporation, ultrafiltration centrifugation and other methods, then the purity is further removed by the refining process of affinity chromatography, ion exchange chromatography and gel filtration chromatography, and the purity is verified by SDS-PAGE electrophoresis and HPLC detection, and then the bacteria are removed by filtration, finally the high-purity RNA helicase inhibitor is packaged and stored under appropriate conditions, ensuring the activity and stability of the product, meeting the experimental or pharmaceutical requirements.

[0027] The working principle of the present application is as follows: during operation, the material is delivered to the inside of the extractor 2 through the first conveying pipe 6, and after the delivery of the material is completed, the first motor 4 is used to rotate the first rotating rod 8, the rotation of the first rotating rod 8 drives the first stirring impeller 9 to rotate, and the material in the inside of the extractor 2 is stirred and extracted, after the extraction treatment, the liquid material needed is delivered to the inside of the washing device 3 through the liquid pump 10, and the liquid material not needed is discharged through the first discharging pipe 7, the liquid material enters the inside of the washing device 3 to be washed, and the material after the extraction treatment does not need to be transferred and can be directly delivered to the washing device 3 for washing through the liquid pump 10; the rotation of the first rotating rod 8 drives the scraping plate 21 to rotate, the rotation of the scraping plate 21 can scrape the material on the inner side wall of the extractor 2, so as to avoid that the material is adsorbed on the inner side wall of the extractor 2 and also to prevent the waste of the material and the failure of the stirring treatment of the material; the rotation of the first stirring impeller 9 can make the material pass through the first layering hole plate 14, the inside of the first layering hole plate 14 is provided with a plurality of holes, and the first layering hole plate 14 is equivalent to a “fine hole sieve”, when the liquid material passes through the small holes of the first layering hole plate 14, the liquid material will be forcibly separated into small droplets, the water phase and the organic phase in the small droplets originally have a tendency to separate, after passing through the small holes, the “clustering” force is lost, and the small droplets will quickly re-aggregate into two layers, the layering speed can be obviously accelerated, and the effect of the layering of the material can be improved through the first layering hole plate 14; the rotation of the first rotating rod 8 drives the first stirring impeller 9 to rotate and stir the material in the inside of the extractor 2, the rotation of the first rotating rod 8 drives the motor 22 and the third rotating rod 23 to rotate together, the motor 22 drives the third rotating rod 23 to rotate, the rotation of the third rotating rod 23 drives the stirring blade 24 to rotate, the rotation direction of the stirring blade 24 is different from that of the first stirring impeller 9, different stirring paths can be generated, the stirring effect on the material is improved, the rotation of the first rotating rod 8 drives the swing blade 25 to rotate above the magnetic plate 26, the end of the swing blade 25 is provided with an iron block, the swing blade 25 is attracted by the magnetic attraction of the magnetic plate 26, so that the swing blade 25 is bent and moves, a plurality of magnetic plates 26 with different positions are arranged on the first layering hole plate 14, the swing blade 25 can change the bending condition several times when rotating one circle on the first layering hole plate 14, the bending of the swing blade 25 cooperates with the rotation of the third rotating rod 23 to change the stirring path and generate different stirring effects, a single stirring direction is easy to form a liquid dead zone in the corners and the center area of the equipment, the contact between the raw material liquid and the extractant is insufficient, the convection formed by different stirring directions can impact and entrap the dead zone liquid, so that the two phases form a three-dimensional circulating mixing in the equipment, the non-contact area is greatly reduced, the target inhibitor can be quickly and fully transferred to the solvent phase, and the washing liquid can react with the residual impurities efficiently.The inside of the heater 18 is provided with a water pump and a heater, which heats water according to the requirement of the extractor 2. The hot water flows into the inside of the heating cavity 20 through the circulating pipe 19, and then flows into the inside of the heater 18, continuously heating the water and entering the inside of the heating cavity 20, thereby heating the extractor 2 according to the requirement. The material enters the inside of the washing device 3 through the liquid pump 10, and the washing liquid enters the inside of the washing device 3 through the second conveying pipe 27. The second motor 5 drives the second rotating rod 11 to rotate, and the rotation of the second rotating rod 11 drives the second stirring impeller 12 to rotate and stir, thereby washing the material in the washing device 3. The rotation of the second rotating rod 11 drives the uniform blade 17 to rotate and stir the bottom of the washing device 3, and then the material is discharged through the second discharge pipe 13. The second layered hole plate 15 is provided with a plurality of holes. When the liquid material passes through the small holes of the second layered hole plate 15, it is forcibly divided into small droplets. The water phase and the organic phase in the small droplets can improve the washing effect through the second layered hole plate 15, and the height of the second layered hole plate 15 can be adjusted according to the amount of the material through the air cylinder 16.

[0028] The above describes one embodiment of the present application in detail, but the above description is only a preferred embodiment of the present application, and cannot be considered as limiting the scope of the present application. Any equivalent changes and improvements made according to the scope of the present application should still belong to the scope of the present application.

Claims

1. An RNA helicase inhibitor purification device based on solvent extraction, characterized in that, The system includes a housing (1), an extractor (2), a washer (3), a first motor (4), a first conveying pipe (6), a first discharge pipe (7), a first rotating rod (8), a first stirring impeller (9), and a liquid pump (10). The extractor (2) is fixed inside the housing (1). The washer (3) is fixed inside the housing (1) to the right side of the extractor (2). The first motor (4) is fixed to the top of the extractor (2). The first rotating rod (8) is fixed to the output end of the first motor (4). The first conveying pipe (6) is fixed to the top of the extractor (2) to the left side of the first motor (4). The first stirring impeller (9) is fixed to the outer wall of the first rotating rod (8). The first discharge pipe (7) is fixed to the bottom of the extractor (2). The liquid pump (10) is located between the extractor (2) and the washer (3).

2. The RNA helicase inhibitor purification device based on solvent extraction according to claim 1, characterized in that, The extractor (2) has a first layered perforated plate (14) fixedly connected inside, and the first rotating rod (8) is rotatably connected to the first layered perforated plate (14).

3. The RNA helicase inhibitor purification device based on solvent extraction according to claim 2, characterized in that, The top of the washer (3) is fixedly connected to a second motor (5); the output end of the second motor (5) is fixedly connected to a second rotating rod (11); a second stirring impeller (12) is fixedly connected to the second rotating rod (11); a second discharge pipe (13) is fixedly connected to the bottom of the washer (3); uniform blades (17) are fixedly connected to the bottom of the second rotating rod (11); and a second conveying pipe (27) is fixedly connected to the top of the washer (3).

4. The RNA helicase inhibitor purification device based on solvent extraction according to claim 3, characterized in that, A cylinder (16) is fixedly connected to the top of the washing machine (3); a second layered perforated plate (15) is fixedly connected to the bottom of the cylinder (16), and the second layered perforated plate (15) is slidably connected inside the washing machine (3).

5. The RNA helicase inhibitor purification device based on solvent extraction according to claim 4, characterized in that, A motor (22) is fixedly connected to the right side of the first rotating rod (8); a third rotating rod (23) is fixedly connected to the output end of the motor (22); a stirring blade (24) is fixedly connected to the outer side wall of the third rotating rod (23); a swing blade (25) is fixedly connected to the middle outer side wall of the third rotating rod (23); a magnet (26) is fixedly connected to the top of the first layered perforated plate (14), and the magnet (26) is located below the swing blade (25).

6. The RNA helicase inhibitor purification device based on solvent extraction according to claim 5, characterized in that, A scraper plate (21) is fixedly connected to the right side of the first rotating rod (8), and the scraper plate (21) is rotatably connected to the inside of the extractor (2).

7. The RNA helicase inhibitor purification device based on solvent extraction according to claim 6, characterized in that, A heater (18) is fixed to the inner wall of the housing (1); a circulation pipe (19) is fixed to the side end of the heater (18); a heating chamber (20) is opened inside the extractor (2), and the bottom end of the circulation pipe (19) is provided in the heating chamber (20).

8. A solvent extraction-based RNA helicase inhibitor purification process, applicable to the RNA helicase inhibitor purification apparatus described in claim 1, characterized in that, Includes the following steps: S1. The raw material containing RNA helicase inhibitor is pretreated by high-pressure homogenization, ultrasonic disruption or enzymatic hydrolysis to destroy cell structure, or by dissolving and filtering with organic solvent to remove solid impurities, and then by centrifugation and membrane filtration to obtain a clear crude extract; then the crude extract is injected into the extractor (2) of the shell (1), and a suitable extractant is added in proportion, the pH is adjusted to near the isoelectric point of the inhibitor and the salt concentration is optimized, and the two phases are fully mixed by multi-directional coordinated stirring in the device, so that the target inhibitor is transferred from the aqueous phase to the organic phase, and the initial separation is completed; S2. After extraction, the organic phase containing the target inhibitor does not need to be transferred. It enters the scrubber (3) directly through the internal flow guide structure of the device and comes into contact with the preset washing liquid. By utilizing the device's differentiated stirring intensity and direction design, while ensuring that the washing liquid and the organic phase are in full contact and efficiently removing residual salts, RNase, endotoxins and other impurities, the emulsification phenomenon is suppressed and the two phases are accelerated to separate. The separated washing liquid containing impurities is discharged through a dedicated outlet, while the organic phase is retained in the device for subsequent processing. The entire process achieves seamless connection between extraction and washing, avoiding pollution and loss caused by intermediate transfer. S3. The washed organic phase is transferred to the aqueous phase by back-extraction or directly sent to the concentration unit. The target product is concentrated and desalted by methods such as vacuum evaporation, ultrafiltration and centrifugation. Then, a purification process using a combination of affinity chromatography, ion exchange chromatography and gel filtration chromatography is used to further remove trace impurities, polymers and residual solvents. After the purity is verified by SDS-PAGE electrophoresis and HPLC detection, sterile filtration is performed. Finally, the high-purity RNA helicase inhibitor is aliquoted and stored under appropriate conditions to ensure the activity and stability of the product and meet experimental or pharmaceutical needs.

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