Gemcitabine liposome preparation all-in-one machine and method

Abstract

The invention provides a gemcitabine lipidosome preparation all-in-one machine and method, the gemcitabine lipidosome preparation all-in-one machine comprises an instrument shell, an operation panel, a microfluidic LNP preparation instrument, a test tube booster, a centrifugal machine and a PH detector, the operation panel is installed on the instrument shell, the microfluidic LNP preparation instrument, the test tube booster, the centrifugal machine and the PH detector are all installed in the instrument shell, and the operation panel is installed on the instrument shell. The micro-fluidic LNP preparation instrument is installed at the topmost part of the instrument shell, the right side of the micro-fluidic LNP preparation instrument is connected with the PH detector, the test tube booster is installed below the micro-fluidic LNP preparation instrument and the PH detector, the centrifugal machine is installed below the test tube booster, and integrated preparation of gemcitabine lipidosome is achieved through the structure.

Description

Technical Field

[0001] The invention belongs to the technical field of drug preparation, and in particular relates to an integrated machine and method for preparing gemcitabine liposomes. Background Art

[0002] The traditional preparation process of gemcitabine liposomes has the problems of complex preparation process, multiple and scattered equipment, requiring a lot of manpower operation and equipment conversion, which is time-consuming, labor-intensive and has high equipment costs. Summary of the invention

[0003] The purpose of the present invention is to overcome the above-mentioned shortcomings and provide an integrated machine and method for preparing gemcitabine liposomes. The integrated machine can be used for the continuous and automated preparation of gemcitabine liposomes, replacing the traditional dispersed preparation process, and can achieve precise control of drug particle size and distribution, improve drug encapsulation rate and stability, thereby enhancing the efficacy of the drug and reducing side effects, and achieving controllable process parameters in the continuous preparation process, with good repeatability, achieving efficient and stable production and preparation, and achieving assembly line and automation of the entire production and preparation process.

[0004] In order to achieve the above-mentioned technical features, the purpose of the present invention is achieved as follows: a gemcitabine liposome preparation all-in-one machine, including an instrument housing, an operation panel, a microfluidic LNP preparation instrument, a test tube booster, a centrifuge and a pH detector, wherein the operation panel is installed on the instrument housing and is used to realize human-computer interaction; the microfluidic LNP preparation instrument, the test tube booster, the centrifuge and the pH detector are all installed inside the instrument housing; wherein the microfluidic LNP preparation instrument is installed at the top of the instrument housing, and a pH detector for monitoring the pH value is connected to one side of the microfluidic LNP preparation instrument; the test tube booster is installed below the microfluidic LNP preparation instrument and the pH detector; the centrifuge is installed below the test tube booster.

[0005] The instrument housing comprises a sliding door, a handle and a sliding slot, the sliding door is installed on the instrument housing through the sliding slot, and the sliding door is equipped with a handle; The operation panel is located on one side of the sliding door and is used to control the operation of the microfluidic LNP preparation instrument, the test tube booster, the centrifuge, and display the detection parameters of the pH tester.

[0006] The microfluidic LNP preparation instrument comprises a high-pressure delivery pump, which is respectively connected to one end of an aqueous phase microfluidic delivery tube, an organic phase microfluidic delivery tube, a weak alkaline solution microfluidic delivery tube and an aqueous solution microfluidic delivery tube; the other ends of the aqueous phase microfluidic delivery tube, the organic phase microfluidic delivery tube, the weak alkaline solution microfluidic delivery tube and the aqueous solution microfluidic delivery tube are respectively connected to an aqueous phase bottle, an organic phase bottle, a weak alkaline solution bottle and an aqueous solution bottle; The microfluidic chip is connected to a preparation pump and a high-pressure delivery pump, and the aqueous phase and the organic phase are delivered into the microfluidic chip at a constant speed in a certain proportion for mixing and emulsification.

[0007] One end of the solution outlet tube is connected to the weak alkaline solution bottle, and the other end is connected to the test tube booster. It is used to add weak alkaline solution after the blank liposomes are prepared by mixing the aqueous phase and the organic phase, so that the drug is stably encapsulated in the aqueous phase of the blank liposomes.

[0008] The test tube booster comprises a ball bearing, an inner ring, a test tube slide groove, a test tube limit groove, a bracket, a guide tube, a test tube cover storage box, a one-way valve, a test tube cover, a test tube fixing groove, a test tube and a test tube slide groove; The ball bearing is installed in cooperation with the inner ring, and a plurality of evenly distributed test tube fixing grooves are fixed on the inner ring, and a test tube is fixedly placed in each test tube fixing groove.

[0009] The test tube caps are placed in a test tube cap storage box, each batch of test tube cap storage boxes stores test tube caps, a one-way valve is installed on the top of each test tube cap, the one-way valve conducts one-way from the top of the test tube cap to the bottom of the test tube cap, the conduit is divided into four branches, the common end is connected to the solution outlet tube, and the other end is respectively introduced into a test tube cap storage box and passes through the one-way valves of multiple test tube caps.

[0010] The test tube chute, the test tube limiting groove, the bracket and the catheter rotate coaxially, the bracket is connected and fixed to the lower end of the test tube cover storage box, the lower end opening of the test tube cover storage box is concentric with the test tube fixed in the test tube fixing groove, when the bracket together with the test tube chute, the test tube limiting groove, and the catheter coaxially rotate until the bottom ends of the four test tube cover storage boxes fixed on the bracket are respectively concentric with one test tube, the drug-encapsulated liposomes prepared by the microfluidic LNP preparation instrument pass through the catheter, the test tube cover storage box and the test tube cover, and are transported to the four concentric test tubes; The outer end of the test tube slide is concentric with the test tube, and the inner end is provided with a test tube lower slide. When the liposomes loaded with drugs are injected into the test tube, the test tube cover storage box seals the test tube with the test tube cover, and the test tube slide is used to shrink four test tubes sealed with the test tube cover along the test tube limiting groove to the test tube lower slide.

[0011] The centrifuge comprises a rotor body, a built-in condensation water tank, a test tube tank and a drum; The rotor body cooperates with the drum through a built-in condensation water tank. Four rotor bodies are evenly distributed in the separation chamber of the centrifuge. Each rotor body has a test tube slot. When the test tube is delivered to the centrifuge through the test tube slide chute, the drum drives the rotor body to rotate so that the test tube slot in the rotor body corresponds to the coaxial center of the test tube, so that the test tube can smoothly enter the test tube slot of the rotor body; The built-in condensation water tank is connected to the rotor body and the drum, and is used to control the temperature of the test tube placed in the rotor body after the ultrafiltration is completed.

[0012] The pH detector includes an electrode holder and an electrode. The electrode holder is installed on the instrument housing, has multiple slots and can rotate for convenient placement of the electrode. The tail end of the electrode is connected to the microfluidic LNP preparation instrument through a wire.

[0013] A method for preparing gemcitabine liposomes using an integrated gemcitabine liposome preparation machine comprises the following steps: S1: Hold the handle and open the sliding door along the slide groove on the instrument housing; S2: Add aqueous phase, organic phase, weak alkaline solution and aqueous solution into the aqueous phase bottle, organic phase bottle, weak alkaline solution bottle and aqueous solution bottle respectively; S3: four test tube caps are placed in four test tube cap storage boxes respectively, the one-way valves on the test tube caps are opened, and the catheter is passed through the test tube caps from the valve openings of the one-way valves to connect the four test tube caps in series; S4: sixteen test tubes are stored in the sixteen test tube limiting slots; S5: Use the operation panel to independently set the parameters of the microfluidic LNP preparation instrument, centrifuge, and pH detector; S6: After preparation, start the equipment and start the preparation of gemcitabine liposomes; S7: The microfluidic LNP preparation instrument controls the preparation pump and the high-pressure delivery pump to draw the aqueous phase and the organic phase from the aqueous phase bottle and the organic phase bottle at a certain ratio and constant speed through the aqueous phase microfluidic delivery tube and the organic phase microfluidic delivery tube respectively; S8: After the aqueous phase and the organic phase enter the microfluidic chip, they are mixed and emulsified. The microfluidic chip achieves a turbulent state of the aqueous phase and the organic phase in the microfluidic chip through a specific flow channel structure and speed control, thereby preparing blank liposomes; S9: The microfluidic LNP preparation instrument controls the preparation pump and the high-pressure delivery pump to deliver the aqueous solution from the aqueous solution bottle to the microfluidic chip, and the blank liposomes are mixed with the aqueous solution to obtain a blank liposome mixed solution; S10: After the system detects that the microfluidic LNP preparation instrument has completed solution preparation, the inner ring of the test tube booster is controlled to drive the test tube slide, the test tube limit groove, the bracket, and the guide tube to rotate through the ball bearing, so that the four test tube cover storage boxes connected to the bracket are respectively aligned with the four test tubes fixed on the test tube fixing groove; S11: The blank liposome mixed solution flows into the catheter through the solution outlet tube and is introduced into the four test tubes along the catheter; S12: The test tube cap storage box matches the test tube caps at the bottom layer with four test tubes respectively, so that the test tubes are sealed; S13: The test tube slideway contracts to make the test tube detach from the test tube fixing groove, and the bottom of the test tube slides along the test tube limiting groove toward the center of the instrument under the thrust of the test tube slideway until the test tube enters the lower test tube slideway; S14: At this time, the centrifuge drum drives the rotor body to rotate, so that the test tube groove in the rotor body is aligned with the lower groove of the test tube; S15: The test tube enters the test tube slot of the rotor body of the centrifuge along the test tube slide groove; S16: The centrifuge is started, and the drum drives the rotor body and the test tubes placed inside the test tube slot to rotate according to the set parameters, and the blank liposome mixed solution inside the test tube is ultrafiltered. During the ultrafiltration process, the one-way valve prevents the blank liposome mixed solution in the test tube from flowing out to ensure sealing; S17: After the ultrafiltration is completed, the test tube is taken out and placed in the test tube fixing tank. The microfluidic LNP preparation instrument controls the preparation pump and the high-pressure delivery pump to deliver the weak alkaline solution from the weak alkaline solution bottle to the test tube, so that the blank liposomes are mixed with the weak alkaline solution, promoting the drug to enter the blank liposomes and increasing the internal and external potential difference; S18: After each pH adjustment, the pH tester is used to adjust and test the concentration of the solution in the test tube; S19: until the drug encapsulation reaches a stable state, the drug is stably encapsulated in the liposome, and then the test tube is put back into the test tube fixing tank; S20: Repeat S13-S15, the test tube enters the test tube tank of the centrifuge again, and the built-in condensation water tank controls the temperature of the solvent in the test tube; S21: First, incubate the solvent in the test tube at 40-50°C for 50 minutes. After incubation, cool it down to 0°C, then start the centrifuge and perform ultrafiltration for another half an hour. The supernatant obtained after the end is the finished product.

[0014] The present invention has the following beneficial effects: 1. A gemcitabine liposome preparation machine integrates microfluidic preparation, solution addition, ultrafiltration of blank liposome mixed solution, temperature control, pH detection and other operation steps, which can achieve precise control of drug particle size and distribution, improve drug encapsulation rate and stability, thereby enhancing drug efficacy and reducing side effects, and achieving controllable process parameters and good repeatability in the continuous preparation process, achieving efficient and stable production and preparation, and realizing assembly line and automation of the entire production and preparation process, changing the situation of traditional complex preparations with high technical difficulty, complex preparation process and large manpower consumption, and helping high-end preparations to meet international advanced quality standards.

[0015] 2. Setting up multiple solution bottles and corresponding microfluidic delivery tubes can realize automatic liquid addition and precise control of solution dosage, making the liquid addition operation more flexible and shortening the preparation time.

[0016] 3. Through the test tube booster, the equipment can automatically complete the filling, sealing and feeding of the test tube into the centrifuge, avoiding manual operation and realizing the upper and lower linkage of the microfluidic LNP preparation instrument and the centrifuge. In addition, the test tube booster can load sixteen test tubes and sixteen test tube caps at a time, and a batch can prepare four medicines at the same time, which improves the preparation efficiency.

[0017] 4. The test tube cap is equipped with a one-way valve to control the one-way flow of the solution in the test tube, so that the solution can only flow into the test tube but not out of the test tube, thereby realizing the automatic liquid addition and sealing of the test tube. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0019] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0020] Figure 2 It is a schematic diagram of the overall internal structure of the present invention.

[0021] Figure 3 It is a schematic diagram of the overall structure of the test tube booster of the present invention.

[0022] Figure 4 It is a schematic diagram of the test tube booster and centrifuge structure of the present invention.

[0023] Figure 5 For the present invention Figure 4 AA section view of the test tube booster and centrifuge.

[0024] Figure 6 It is a schematic diagram of the three-dimensional structure of the test tube cover of the present invention.

[0025] Figure 7 It is a schematic diagram of the structure of the centrifuge of the present invention.

[0026] In the figure: instrument housing 1, sliding door 101, handle 102, sliding slot 103; Operation panel 2; Microfluidic LNP preparation instrument 3, microfluidic chip 301, aqueous phase microfluidic delivery tube 302, organic phase microfluidic delivery tube 303, aqueous solution bottle 304, aqueous solution microfluidic delivery tube 305, aqueous phase bottle 306, organic phase bottle 307, weak alkaline solution bottle 308, weak alkaline solution microfluidic delivery tube 309, solution outlet tube 310; Test tube booster 4, ball bearing 401, inner ring 402, test tube slide groove 403, test tube limit groove 404, bracket 405, guide tube 406, test tube cover storage box 407, one-way valve 408, test tube cover 409, test tube fixing groove 410, test tube 411, and test tube lower slide groove 412; Centrifuge 5, rotor body 501, built-in condensation water tank 502, test tube tank 503, drum 504; PH detector 6, electrode holder 601, electrode 602. DETAILED DESCRIPTION

[0027] The embodiments of the present invention are further described below in conjunction with the accompanying drawings.

[0028] Embodiment 1: See also Figure 1-7 A gemcitabine liposome preparation integrated machine includes an instrument housing 1, an operation panel 2, a microfluidic LNP preparation instrument 3, a test tube booster 4, a centrifuge 5 and a pH detector 6, wherein the operation panel 2 is installed on the instrument housing 1, the microfluidic LNP preparation instrument 3, the test tube booster 4, the centrifuge 5 and the pH detector 6 are all installed inside the instrument housing 1, the microfluidic LNP preparation instrument 3 is installed at the top of the instrument housing 1, the pH detector 6 is connected to the right side of the microfluidic LNP preparation instrument 3, the test tube booster 4 is installed below the microfluidic LNP preparation instrument 3 and the pH detector 6, and the centrifuge 5 is installed below the test tube booster 4.

[0029] See also Figure 1 , Figure 2 The instrument housing 1 includes a sliding door 101, a handle 102 and a slide groove 103. The sliding door 101 is installed on the instrument housing 1 through the slide groove 103. The sliding door 101 is equipped with a handle 102. The internal structure of the gemcitabine liposome preparation all-in-one machine can be seen by holding the handle 102 and opening the sliding door 101 along the slide groove 102, so as to facilitate the replacement and replenishment of the test tube 411, the test tube cover 409, the aqueous phase solution in the aqueous phase bottle 306, the organic phase solution in the organic phase bottle 307, the weak alkaline solution in the weak alkaline solution bottle 308 and the aqueous solution in the aqueous solution bottle 304 during the gemcitabine liposome preparation process.

[0030] See also Figure 1 The operation panel 2 is installed on the operation housing 1 and is located on the left side of the sliding door 101. It can be used to control the operation of the microfluidic LNP preparation instrument 3, the test tube booster 4, and the centrifuge 5 and display the detection parameters of the pH detector.

[0031] See also Figure 2 The microfluidic LNP preparation instrument 3 includes a preparation pump, a high-pressure delivery pump, a microfluidic chip 301, an aqueous phase bottle 306, an aqueous phase microfluidic delivery tube 302, an organic phase bottle 307, an organic phase microfluidic delivery tube 303, a weak alkaline solution bottle 308, a weak alkaline solution microfluidic delivery tube 309, an aqueous solution bottle 304, an aqueous solution microfluidic delivery tube 305 and a solution outlet tube 310.

[0032] Preferably, the microfluidic LNP preparation instrument 3 adopts microfluidic technology, based on fluid mechanics theory, to realize the preparation and processing technology of blank liposomes in the pipeline, which can realize the functions of sample mixing, emulsification, separation and purification, etc. The parameters of blank liposomes are controllable in the continuous preparation process, and have good reproducibility, so it is more scalable than the traditional step-by-step split preparation and batch production methods. It can realize the functions of primary emulsification, secondary emulsification and particle size control of samples.

[0033] Specifically, the high-pressure delivery pump in the microfluidic LNP preparation instrument 3 is respectively connected to one end of the aqueous phase microfluidic delivery tube 302, the organic phase microfluidic delivery tube 303, the weak alkaline solution microfluidic delivery tube 309 and the aqueous solution microfluidic delivery tube 305, and the other ends of the aqueous phase microfluidic delivery tube 302, the organic phase microfluidic delivery tube 303, the weak alkaline solution microfluidic delivery tube 309 and the aqueous solution microfluidic delivery tube 305 are respectively connected to the aqueous phase bottle 306, the organic phase bottle 307, the weak alkaline solution bottle 308 and the aqueous solution bottle 304.

[0034] Specifically, the microfluidic chip 301 is connected to the preparation pump and the high-pressure delivery pump, and the aqueous phase and the organic phase are delivered into the microfluidic chip 301 at a constant rate in a certain ratio for mixing and emulsification.

[0035] Furthermore, the volume of the channel in the microfluidic chip 301 is extremely small. The flow rate of the aqueous phase and the organic phase liquid can be precisely controlled by the microfluidic LNP preparation instrument 3, so that the two-phase liquid can undergo a rapid self-assembly reaction at a very small phase interface in the form of laminar flow, and the reaction time is usually within 3ms. The microfluidic chip 301 is like a precise reactor for microquantification, which can quickly obtain lipid nanoparticles with uniform size and good repeatability by controlling and intensifying the mixing of the fluid. The method has mild preparation conditions, fast speed, and is easy to scale up. The morphology, structure, and particle size distribution of the synthesized nanoparticles are uniform, and the batch-to-batch repeatability is controllable.

[0036] See also Figure 2 One end of the solution outlet tube 310 is connected to the weak alkaline solution bottle 308, and the other end is connected to the test tube booster 4, which is used to add the weak alkaline solution after the blank liposomes are prepared by mixing the aqueous phase and the organic phase, so that the drug can be stably encapsulated in the aqueous phase of the blank liposomes.

[0037] See also Figure 2 , Figure 3 , Figure 4 , 5 The test tube booster 4 includes a ball bearing 401, an inner ring 402, a test tube slide groove 403, a test tube limiting groove 404, a bracket 405, a guide tube 406, a test tube cover storage box 407, a one-way valve 408, a test tube cover 409, a test tube fixing groove 410, a test tube 411, and a test tube lower slide groove 412.

[0038] Specifically, the ball bearing 401 is installed in cooperation with the inner ring 402, and sixteen evenly distributed test tube fixing grooves 410 are fixed on the inner ring 402, and a test tube 411 is fixedly placed in each test tube fixing groove 410. The test tube caps 409 are placed in the test tube cap storage box 407, and four test tube caps 409 are stored in each batch of test tube cap storage boxes 407. A one-way valve 408 is installed on the top of each test tube cap 409, and the one-way valve 408 is one-way from the top of the test tube cap 409 to the bottom of the test tube cap 409. The conduit 406 is divided into four branches, and the common end is connected to the solution outlet tube 310, and the other end is respectively introduced into a test tube cap storage box 407, and passes through the one-way valves 408 of the four test tube caps 409. The test tube slide groove 403, the test tube limiting groove 404, the bracket 405 and the catheter 406 rotate coaxially, and the bracket 405 is connected and fixed to the lower end of the test tube cover storage box 407. The lower end opening of the test tube cover storage box 407 is concentric with the test tube 411 fixed in the test tube fixing groove 410. When the bracket 405 together with the test tube slide groove 403, the test tube limiting groove 404 and the catheter 406 rotate coaxially until the bottom ends of the four test tube cover storage boxes 407 fixed on the bracket 405 are respectively concentric with one test tube 411, the drug-encapsulated liposomes prepared by the microfluidic LNP preparation instrument 3 can pass through the catheter 406, pass through the test tube cover storage box 407 and the test tube cover 409, and be transported to the four concentric test tubes 411.

[0039] Furthermore, the outer end of the test tube slide 403 is concentric with the test tube 411, and the inner end has a test tube lower slide 412. After the liposomes loaded with drugs are injected into the test tube 411, the test tube cover storage box 407 seals the test tube 411 with the test tube cover 409, and the test tube slide 403 is used to shrink the four test tubes 411 sealed with the test tube cover 409 along the test tube limiting groove 404 to the test tube lower slide 412.

[0040] See also Figure 7 The centrifuge 5 is located directly below the test tube booster 4 and includes a rotor body 501 , a built-in condensation water tank 502 , a test tube tank 503 , and a drum 504 .

[0041] Specifically, the rotor body 501 cooperates with the drum 504 through the built-in condensation water tank 502, and the four rotor bodies 501 are evenly distributed in the separation chamber of the centrifuge 5. Each rotor body 501 has a test tube slot 503. When the test tube 411 is delivered to the centrifuge 5 through the test tube lower slide 412, the drum 504 drives the rotor body 501 to rotate so that the test tube slot 503 in the rotor body 501 corresponds to the test tube 411 coaxially, so that the test tube 411 can smoothly enter the test tube slot 503 of the rotor body 501. The built-in condensation water tank 502 is connected to the rotor body 501 and the drum 504, and is used to control the temperature of the test tube 411 placed in the rotor body 501 after ultrafiltration is completed.

[0042] See also Figure 2The pH detector 6 is located on the right side of the microfluidic LNP preparation instrument 3, and includes an electrode holder 601 and an electrode 602. The electrode holder 601 is installed on the instrument housing 1, has multiple slots and is rotatable for conveniently placing the electrode 602. The tail end of the electrode 602 is connected to the microfluidic LNP preparation instrument 3 through a wire.

[0043] Embodiment 2: A method for using a gemcitabine liposome preparation machine, which is used for preparing gemcitabine liposomes, comprises the following steps: S1: Hold the handle 102 and open the sliding door 101 along the sliding groove 103 on the instrument housing 1; S2: Adding aqueous phase, organic phase, weak alkaline solution and aqueous solution into the aqueous phase bottle 306, organic phase bottle 307, weak alkaline solution bottle 308 and aqueous solution bottle 304 respectively; S3: four test tube covers 409 are placed in four test tube cover storage boxes 407 respectively, the one-way valves 408 on the test tube covers 409 are opened, and the guide tube 406 passes through the test tube covers 409 from the valve opening of the one-way valve 408, so as to connect the four test tube covers 409 in series; S4: sixteen test tubes 411 are stored in the sixteen test tube limiting slots 404; S5: Use the operation panel 2 to independently set the parameters of the microfluidic LNP preparation instrument 3, the centrifuge 5, and the pH detector 6; S6: After preparation, start the equipment and start the preparation of gemcitabine liposomes; S7: The microfluidic LNP preparation instrument 3 controls the preparation pump and the high-pressure delivery pump to draw the aqueous phase and the organic phase from the aqueous phase bottle 306 and the organic phase bottle 307 at a certain ratio and constant speed through the aqueous phase microfluidic delivery tube 302 and the organic phase microfluidic delivery tube 303 respectively; S8: After the aqueous phase and the organic phase enter the microfluidic chip 301, they are mixed and emulsified. The microfluidic chip 301 achieves a turbulent state of the aqueous phase and the organic phase in the microfluidic chip 301 through a specific flow channel structure and speed control, thereby preparing blank liposomes; S9: the microfluidic LNP preparation instrument 3 controls the preparation pump and the high-pressure delivery pump to deliver the aqueous solution from the aqueous solution bottle 304 to the microfluidic chip 301, and the blank liposomes are mixed with the aqueous solution to obtain a blank liposome mixed solution; S10: After the system detects that the microfluidic LNP preparation instrument 3 has completed the solution preparation, the system controls the inner ring 402 of the test tube booster 4 to drive the test tube slide 403, the test tube limit groove 404, the bracket 405, and the guide tube 406 to rotate through the ball bearing 401, so that the four test tube cover storage boxes 407 connected to the bracket 405 are respectively aligned with the four test tubes 411 fixed on the test tube fixing groove 410; S11: The blank liposome mixed solution flows into the conduit 406 through the solution outlet tube 310 and is introduced into the four test tubes 411 along the conduit 406; S12: The test tube cap storage box 407 matches the test tube caps 409 at the bottom with four test tubes 411 respectively, so that the test tubes 411 are sealed; S13: the test tube slide groove 403 contracts to make the test tube 411 detach from the test tube fixing groove 410, and the bottom of the test tube 411 slides along the test tube limiting groove 404 toward the center of the instrument under the contraction thrust of the test tube slide groove 403, until the test tube 411 enters the lower test tube slide groove 412; S14: At this time, the drum 504 of the centrifuge 5 drives the rotor body 501 to rotate, so that the test tube slot 503 in the rotor body 501 is aligned with the lower groove 412 of the test tube; S15: the test tube 411 enters the test tube slot 503 of the rotor body 501 of the centrifuge 5 along the test tube lower groove 412; S16: The centrifuge 5 is started, and the drum 504 drives the rotor body 501 and the test tube 411 placed inside the test tube slot 503 to rotate according to the set parameters, and the blank liposome mixed solution inside the test tube 411 is ultrafiltered. During the ultrafiltration process, the one-way valve 408 prevents the blank liposome mixed solution in the test tube 411 from flowing out to ensure sealing; S17: After the ultrafiltration is completed, the test tube 411 is taken out and placed in the test tube fixing tank 410, and the microfluidic LNP preparation instrument 3 controls the preparation pump and the high-pressure delivery pump to deliver the weak alkaline solution from the weak alkaline solution bottle 308 to the test tube 411, so that the blank liposomes are mixed with the weak alkaline solution, promoting the drug to enter the blank liposomes, and increasing the internal and external potential difference; S18: After each pH adjustment, the pH detector 6 is used to adjust and detect the concentration of the solution in the test tube 411; S19: until the drug encapsulation reaches a stable state, the drug is stably encapsulated in the liposome, and then the test tube 411 is put back into the test tube fixing groove 410; S20: Repeat S13-S15, the test tube 411 enters the test tube tank 503 of the centrifuge 5 again, and the built-in condensation water tank 502 controls the temperature of the solvent in the test tube 411; S21: First, the solvent in the test tube 411 is incubated at a temperature of 40-50°C for 50 minutes. After the incubation, the temperature is lowered to 0°C, and then the centrifuge 5 is started to ultrafilter again for half an hour. The supernatant obtained after the ultrafiltration is the finished product.

Claims

1. A machine for preparing gemcitabine liposomes, It is characterized in that The apparatus comprises an instrument housing (1), an operation panel (2), a microfluidic LNP preparation instrument (3), a test tube booster (4), a centrifuge (5) and a pH detector (6), wherein the operation panel (2) is mounted on the instrument housing (1) and is used to realize human-computer interaction; the microfluidic LNP preparation instrument (3), the test tube booster (4), the centrifuge (5) and the pH detector (6) are all mounted inside the instrument housing (1); wherein the microfluidic LNP preparation instrument (3) is mounted on the top of the instrument housing (1), and a pH detector (6) for monitoring the pH value is connected to one side of the microfluidic LNP preparation instrument (3); the test tube booster (4) is mounted below the microfluidic LNP preparation instrument (3) and the pH detector (6); and the centrifuge (5) is mounted below the test tube booster (4).

2. According to claim 1, a machine for preparing gemcitabine liposomes, Features: The instrument housing (1) comprises a sliding door (101), a handle (102) and a sliding groove (103); the sliding door (101) is mounted on the instrument housing (1) via the sliding groove (103); and the handle (102) is mounted on the sliding door (101); The operation panel (2) is located on one side of the sliding door (101) and is used to control the operation of the microfluidic LNP preparation instrument (3), the test tube booster (4), and the centrifuge (5) and to display the detection parameters of the pH detector (6).

3. The all-in-one machine for preparing gemcitabine liposomes according to claim 2, Features: The microfluidic LNP preparation apparatus (3) comprises a high-pressure delivery pump, which is respectively connected to one end of an aqueous phase microfluidic delivery tube (302), an organic phase microfluidic delivery tube (303), a weak alkaline solution microfluidic delivery tube (309) and an aqueous solution microfluidic delivery tube (305); the other ends of the aqueous phase microfluidic delivery tube (302), the organic phase microfluidic delivery tube (303), the weak alkaline solution microfluidic delivery tube (309) and the aqueous solution microfluidic delivery tube (305) are respectively connected to an aqueous phase bottle (306), an organic phase bottle (307), a weak alkaline solution bottle (308) and an aqueous solution bottle (304); The microfluidic chip (301) is connected to a preparation pump and a high-pressure delivery pump, and the aqueous phase and the organic phase are delivered into the microfluidic chip (301) at a constant speed in a certain ratio for mixing and emulsification.

4. The all-in-one machine for preparing gemcitabine liposomes according to claim 3, Features: One end of the solution outlet tube (310) is connected to the weak alkaline solution bottle (308), and the other end is connected to the test tube booster (4). It is used to add the weak alkaline solution after the blank liposomes are prepared by mixing the aqueous phase and the organic phase, so that the drug is stably encapsulated in the aqueous phase of the blank liposomes.

5. A machine for preparing gemcitabine liposomes according to claim 4, Features: The test tube booster (4) comprises a ball bearing (401), an inner ring (402), a test tube slide groove (403), a test tube limiting groove (404), a bracket (405), a guide tube (406), a test tube cover storage box (407), a one-way valve (408), a test tube cover (409), a test tube fixing groove (410), a test tube (411) and a test tube lower slide groove (412); The ball bearing (401) is installed in cooperation with the inner ring (402). The inner ring (402) is fixed with a plurality of evenly distributed test tube fixing grooves (410), and a test tube (411) is fixedly placed in each test tube fixing groove (410).

6. The all-in-one machine for preparing gemcitabine liposomes according to claim 5, Features: The test tube caps (409) are placed in a test tube cap storage box (407), and each batch of test tube cap storage boxes (407) stores test tube caps (409). A one-way valve (408) is installed on the top of each test tube cap (409), and the one-way valve (408) is unidirectionally connected from the top of the test tube cap (409) to the bottom of the test tube cap (409). The conduit (406) is divided into four branches, and the common end is connected to the solution outlet tube (310), and the other end is respectively introduced into a test tube cap storage box (407) and passes through the one-way valves (408) of multiple test tube caps (409).

7. The all-in-one machine for preparing gemcitabine liposomes according to claim 6, Features: The test tube slide groove (403), the test tube limiting groove (404), the bracket (405) and the conduit (406) rotate coaxially, the bracket (405) is connected and fixed to the lower end of the test tube cover storage box (407), and the lower end opening of the test tube cover storage box (407) is concentric with the test tube (411) fixed in the test tube fixing groove (410). When the bracket (405) together with the test tube slide groove (403), the test tube limiting groove (404) and the conduit (406) rotate coaxially until the bottom ends of the four test tube cover storage boxes (407) fixed on the bracket (405) are respectively concentric with one test tube (411), the drug-encapsulated liposomes prepared by the microfluidic LNP preparation device (3) pass through the conduit (406), the test tube cover storage box (407) and the test tube cover (409), and are transported to the four concentric test tubes (411); The outer end of the test tube slide groove (403) is concentric with the test tube (411), and the inner end is provided with a test tube lower groove (412). After the liposomes loaded with drugs are injected into the test tube (411), the test tube cover storage box (407) seals the test tube (411) with the test tube cover (409). The test tube slide groove (403) is used to retract the four test tubes (411) sealed with the test tube covers (409) along the test tube limiting groove (404) to the test tube lower groove (412).

8. The all-in-one machine for preparing gemcitabine liposomes according to claim 7, Features: The centrifuge (5) comprises a rotor body (501), a built-in condensation water tank (502), a test tube tank (503) and a rotating drum (504); The rotor body (501) cooperates with the drum (504) via a built-in condensation water tank (502), and the four rotor bodies (501) are evenly distributed in the separation chamber of the centrifuge (5). Each rotor body (501) has a test tube slot (503). When the test tube (411) is delivered into the centrifuge (5) through the test tube lower slide groove (412), the drum (504) drives the rotor body (501) to rotate so that the test tube slot (503) in the rotor body (501) corresponds to the test tube (411) coaxially, so that the test tube (411) can smoothly enter the test tube slot (503) of the rotor body (501); The built-in condensation water tank (502) is connected to the rotor body (501) and the drum (504) and is used to control the temperature of the test tube (411) placed in the rotor body (501) after ultrafiltration is completed.

9. The all-in-one machine for preparing gemcitabine liposomes according to claim 8, Features: The pH detector (6) comprises an electrode holder (601) and an electrode (602). The electrode holder (601) is mounted on the instrument housing (1) and has a plurality of slots and is rotatable for conveniently placing the electrode (602). The tail end of the electrode (602) is connected to the microfluidic LNP preparation instrument (3) via a wire.

10. A method for preparing gemcitabine liposomes by using the all-in-one machine for preparing gemcitabine liposomes according to claim 9, It is characterized in that The following steps are involved: S1: Hold the handle (102) and open the sliding door (101) along the sliding groove (103) on the instrument housing (1); S2: adding an aqueous phase, an organic phase, a weak alkaline solution, and an aqueous solution into the aqueous phase bottle (306), the organic phase bottle (307), the weak alkaline solution bottle (308), and the aqueous solution bottle (304) respectively; S3: four test tube covers (409) are placed in four test tube cover storage boxes (407) respectively, the one-way valves (408) on the test tube covers (409) are opened, and the guide tube (406) passes through the test tube covers (409) from the valve opening of the one-way valve (408), thereby connecting the four test tube covers (409) in series; S4: storing sixteen test tubes (411) in the sixteen test tube limiting slots (404); S5: Use the operation panel (2) to independently set the relevant parameters of the microfluidic LNP preparation instrument (3), the centrifuge (5), and the pH detector (6); S6: After preparation, start the equipment and start the preparation of gemcitabine liposomes; S7: The microfluidic LNP preparation apparatus (3) controls the preparation pump and the high-pressure delivery pump to draw the aqueous phase and the organic phase from the aqueous phase bottle (306) and the organic phase bottle (307) at a constant rate and a certain ratio through the aqueous phase microfluidic delivery tube (302) and the organic phase microfluidic delivery tube (303), respectively; S8: After the aqueous phase and the organic phase enter the microfluidic chip (301), they are mixed and emulsified. The microfluidic chip (301) achieves a turbulent state of the aqueous phase and the organic phase in the microfluidic chip (301) through a specific flow channel structure and speed control, thereby preparing blank liposomes; S9: the microfluidic LNP preparation apparatus (3) controls the preparation pump and the high-pressure delivery pump to deliver the aqueous solution from the aqueous solution bottle (304) to the microfluidic chip (301), and the blank liposomes are mixed with the aqueous solution to obtain a blank liposome mixed solution; S10: After the system detects that the microfluidic LNP preparation device (3) has completed solution preparation, the system controls the inner ring (402) of the test tube booster (4) to drive the test tube slide groove (403), the test tube limit groove (404), the bracket (405), and the guide tube (406) to rotate through the ball bearing (401), so that the four test tube cover storage boxes (407) connected to the bracket (405) are respectively aligned with the four test tubes (411) fixed on the test tube fixing groove (410); S11: The blank liposome mixed solution flows into the conduit (406) through the solution outlet tube (310), and is introduced into the four test tubes (411) along the conduit (406); S12: The test tube cap storage box (407) matches the test tube caps (409) at the bottom layer with four test tubes (411) respectively, so that the test tubes (411) are sealed; S13: the test tube slide groove (403) contracts to cause the test tube (411) to detach from the test tube fixing groove (410), and the bottom of the test tube (411) slides along the test tube limiting groove (404) toward the center of the instrument under the action of the contraction thrust of the test tube slide groove (403) until the test tube (411) enters the lower test tube slide groove (412); S14: At this time, the drum (504) of the centrifuge (5) drives the rotor body (501) to rotate, so that the test tube groove (503) in the rotor body (501) is aligned with the lower groove (412) of the test tube; S15: the test tube (411) enters the test tube slot (503) of the rotor body (501) of the centrifuge (5) along the test tube lower slide groove (412); S16: the centrifuge (5) is started, and the drum (504) drives the rotor body (501) and the test tube (411) placed inside the test tube slot (503) to rotate according to the set parameters, and the blank liposome mixed solution inside the test tube (411) is ultrafiltered. During the ultrafiltration process, the one-way valve (408) prevents the blank liposome mixed solution in the test tube (411) from flowing out to ensure sealing; S17: After the ultrafiltration is completed, the test tube (411) is taken out and placed in the test tube fixing tank (410), and the microfluidic LNP preparation device (3) controls the preparation pump and the high-pressure delivery pump to deliver the weak alkaline solution from the weak alkaline solution bottle (308) to the test tube (411), so that the blank liposomes are mixed with the weak alkaline solution, promoting the drug to enter the blank liposomes, and increasing the internal and external potential difference; S18: After each pH adjustment, the pH detector (6) is used to adjust and detect the concentration of the solution in the test tube (411); S19: until the drug encapsulation reaches a stable state, the drug is stably encapsulated in the liposome, and then the test tube (411) is placed back into the test tube fixing groove (410); S20: Repeat S13-S15, the test tube (411) enters the test tube tank (503) of the centrifuge (5) again, and the built-in condensation water tank (502) controls the temperature of the solvent in the test tube (411); S21: First, the solvent in the test tube (411) is incubated at a temperature of 40-50°C for 50 minutes. After the incubation, the temperature is cooled down to 0°C, and then the centrifuge (5) is started and ultrafiltration is performed again for half an hour. The supernatant obtained after the end is the finished product.

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