JPWO2021227353A5

JPWO2021227353A5 -

Info

Publication number: JPWO2021227353A5
Application number: JP2022568578A
Authority: JP
Publication date: 2023-10-20

Claims

A dispersed phase liquid is caused to flow from one side of the perforated plate to the other side of the perforated plate through a plurality of microwells in the perforated plate, and a continuous phase liquid is caused to flow between the perforated plate and the perforated plate on the other side of the perforated plate. A method of preparing an emulsion comprising forming liquid microspheres in the flowing continuous phase liquid by flowing in parallel and applying shear to the dispersed phase liquid passing through the perforated plate.

The method according to claim 1, wherein the diameter of the microwell is between 0.1 μm and 500 μm.

3. A method according to claim 1 or 2 , wherein the emulsion is a water-in-oil emulsion.

The method according to claim 1 or 2 , wherein the size of the liquid microspheres is adjusted by adjusting the flow rate of the dispersed phase liquid and/or the continuous phase liquid.

Step 1) preparing a dispersed phase liquid and a continuous phase liquid, comprising:
The dispersed phase liquid contains an artificial synthetic polymer and/or a natural biopolymer, and a curing agent,
the continuous phase liquid contains an organic solvent and a nonionic surfactant;
Step 2) The dispersed phase liquid is caused to flow from one side of the perforated plate to the other side of the perforated plate through a plurality of microwells arranged in the perforated plate, and the continuous phase liquid is caused to flow from one side of the perforated plate to the other side of the perforated plate. forming liquid microspheres in the flowing continuous phase liquid by flowing parallel to the perforated plate on the other side and applying shear to the dispersed phase liquid passing through the perforated plate;
step 3) forming particulate matter by reacting the synthetic synthetic polymer and/or natural biopolymer in the liquid microspheres with a curing agent; and step 4) collecting and washing the particulate matter. including that
A method for preparing microcarrier particles, wherein the temperature of the continuous phase liquid in step 2) is 0° C. or lower, and step 3) is performed at 0° C. or lower for 2 to 72 hours.

Step 2) is carried out in a container that includes the perforated plate, the perforated plate divides the interior of the container into a first part and a second part, and the dispersed phase liquid is provided in the container. The continuous phase liquid enters the first portion through a dispersed phase inlet communicating with the first portion, and flows through the perforated plate and enters the second portion. The mixed liquid containing the liquid microspheres after mixing the dispersed phase liquid and the continuous phase liquid enters the second part through the continuous phase inlet communicating with the second part, and the liquid mixture containing the liquid microspheres after mixing the dispersed phase liquid and the continuous phase liquid enters the second part through the continuous phase inlet communicating with the second part. 6. The method of claim 5, wherein the container exits the container through a container outlet in communication with the second portion, the container outlet and the continuous phase inlet being located on opposite sides of the container.

The dispersed phase liquid enters the first part of the vessel by gas pressure and flows through the perforated plate, and the continuous phase liquid enters the second part of the vessel by means of a gear pump and flows through the perforated plate. 6. The method of claim 5 , wherein the flow is parallel to the flow.

6. The method according to claim 5 , wherein step 3) is carried out in a tank equipped with a stirring device.

6. A method according to claim 5 , wherein step 4) is carried out by vacuum suction in a tank in which a filtration device is arranged.

The method according to claim 5 , wherein the diameter of the microwell is between 0.1 μm and 500 μm.

The method according to claim 5 , wherein the flow rate of the continuous phase liquid is 5 to 20 times the flow rate of the dispersed phase liquid at the same time.

The artificially synthesized polymers include polyethylene glycol, polyethylene glycol derivatives, polyethylene glycol diacrylate, polypropylene, polystyrene, polyacrylamide, polylactic acid, polyhydroxy acid, polylactic acid alcoholic acid copolymer, polydimethylsiloxane, polyacid anhydride, and polyacid ester. , polyamide, polyamino acid, polyacetal, polycyanoacrylate, polyurethane, polypyrrole, polyester, polymethacrylate, polyethylene, polycarbonate, and polyethylene oxide ;
Optionally, the natural biopolymer is from at least one of collagen, proteoglycans, glycoproteins, gelatin, gelatin derivatives, chitin, alginate, alginate derivatives, agar, fibrinogen, matrigel, hyaluronic acid, laminin and fibronectin. 6. The method of claim 5, wherein:

The organic solvent is selected from at least one of hydrofluoroether, carbon tetrachloride, petroleum ether, cyclohexane, liquid paraffin, edible oil, soybean oil, olive oil, chloroform, dichloromethane, carbon tetrachloride, and tetrachloroethylene;
Optionally, the nonionic surfactant is sorbitan fatty acid ester, glycerin fatty acid ester, lauric acid ester, polyoxyethylene alkylphenyl ether, polyoxyethylene high carbon fatty alcohol ether, Span, PO-500, monooleic acid ester. and Tween.

The curing agent includes divinylbenzene, diisocyanate, N-hydroxysuccinimide, N,N-methylenebisacrylamide, formaldehyde, glutaraldehyde, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, calcium ion, and tetramethyl. 6. The method of claim 5 , wherein the method is selected from at least one of ethylenediamine, ammonium sulfate, genipin and transglutaminase.

The dispersed phase liquid further includes a buffering agent selected from the group consisting of carboxymethylcellulose, sodium chloride, polyacrylamide, potassium chloride, polyvinylpyrrolidone, sodium sulfate, calcium chloride, sodium chloride, sodium carbonate, and sodium bicarbonate. 6. The method of claim 5 , wherein the method is selected from at least one.

washing the particulate matter with a cleaning agent, the cleaning agent being acetone, anhydrous copper sulfate, calcium chloride, sodium sulfate, absolute ethanol, medical alcohol, hydrofluoroether, sodium alkylbenzene sulfonate, aliphatic alcohol sulfate; 6. The method of claim 5 , wherein the method is selected from at least one of sodium, sodium tripolyphosphate and deionized water.

6. The method of claim 5 , wherein the ratio of organic solvent to nonionic surfactant in the continuous phase liquid is from 5:1 to 20:1 by weight.

1) Container,
2) a perforated plate including a plurality of microwells arranged in the container, the perforated plate dividing the inside of the first container into a first part and a second part;
3) a dispersed phase inlet communicating with the first portion for introducing a dispersed phase liquid;
4) a continuous phase inlet in communication with said second portion for introducing a continuous phase liquid; and 5) a container outlet in communication with said second portion.
An emulsion preparation device comprising: the container outlet and the continuous phase inlet, wherein the continuous phase liquid introduced from the continuous phase inlet passes through the second portion in a direction parallel to the perforated plate. an emulsion preparation device located on the opposite side of the container to allow flow and further flow out of the container outlet;

The emulsion preparation device according to claim 18 , wherein the microwell has a diameter of 0.1 μm to 500 μm.

19. The emulsion preparation device of claim 18 , wherein the continuous phase inlets are two or more and the container outlets are two or more.

19. The emulsion preparation apparatus of claim 18 , wherein the continuous phase inlet and the container outlet have the same horizontal height relative to the bottom of the container.