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WO-2026092305-A1 - MICROFLUIDIC PREPARATION METHOD FOR FAT BODY DELIVERY PLATFORM

WO2026092305A1WO 2026092305 A1WO2026092305 A1WO 2026092305A1WO-2026092305-A1

Abstract

The present invention provides a microfluidic preparation method for a fat body delivery platform, comprising the following steps: S1) providing a lipid solution and an aqueous phase solution, wherein the lipid solution comprises a polar lipid and a non-polar lipid, and the molar ratio of the polar lipid to the non-polar lipid is 1:(0.5-6); and S2) introducing each of the lipid solution and the aqueous phase solution into a microfluidic chip to obtain a fat body delivery platform. Compared with the prior art, the present invention relates to rapidly mixing the lipid solution and the aqueous phase solution in a channel of the microfluidic chip to form the fat body delivery platform, which significantly improves the utilization rate of fat body raw materials, can achieve rapid and efficient preparation of fat bodies having high purity and a uniform particle size, and can also achieve loading of fat-soluble small-molecule drugs, thereby paving the way for industrial production of the fat body delivery platform.

Inventors

  • ZHI, Zelun
  • ZHANG, SHUYAN
  • ZHANG, Gaoxin
  • CAO, Zhen

Assignees

  • 维康平生(北京)生物科技有限公司

Dates

Publication Date
20260507
Application Date
20251024
Priority Date
20241029

Claims (17)

  1. A microfluidic preparation method for a fat body delivery platform, characterized by comprising the following steps: S1) Provide a lipid phase solution and an aqueous phase solution; the lipid phase solution includes polar lipids and nonpolar lipids; the molar ratio of the polar lipids to the nonpolar lipids is (10-50):(50-90); S2) The lipid phase solution and the aqueous phase solution are respectively introduced into the microfluidic chip to obtain the fat body delivery platform.
  2. According to the microfluidic preparation method of claim 1, the solvent of the lipid phase solution is selected from ethanol and/or acetone; the total concentration of polar lipids and nonpolar lipids in the lipid phase solution is 1-60 mM or 2.5-10 mg/mL.
  3. According to the microfluidic preparation method of claim 1, the polar lipid includes one or more of phospholipids, cholesterol, functional polar lipids and cationic lipids. The phospholipids are selected from one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, cardiolipin, and sphingomyelin; The functional polar lipids are selected from one or more of polyethylene glycol modified sterols, biotin modified sterols, amino acid modified sterols, peptide modified sterols, polysaccharide modified sterols, nucleic acid modified sterols, polyethylene glycol modified phospholipids, biotin modified phospholipids, amino acid modified phospholipids, peptide modified phospholipids, polysaccharide modified phospholipids, and nucleic acid modified phospholipids. The cationic lipid is selected from one or more of (2,3-dioleoyl-propyl)-trimethylammonium chloride, (2,3-dioleoyl-propyl)-trimethylamine, 2,3-dioleoyloxy-N-[2-(sperminecarbamoylamino)ethyl]-N,N-dimethyl-1-propylamine hydrochloride, 1,2-dioleoyl-sn-glycerol-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) and 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride; The neutral lipids are selected from one or more of the following: triglycerides, wax esters, sterol esters, sterol esters, retinol esters, ether esters, polyhydroxy fatty acid esters, vitamin A, vitamin D, vitamin E, and vitamin K.
  4. According to claim 1, the microfluidic preparation method is characterized in that the lipid phase solution further includes polyunsaturated fatty acids; the volume ratio of the nonpolar lipid to the polyunsaturated fatty acids is 1:(0.5-2).
  5. According to the microfluidic preparation method of claim 4, the lipid phase solution further includes a drug; the mass ratio of the total mass of the nonpolar lipids and polyunsaturated fatty acids to the mass of the drug is (5-20):1.
  6. The microfluidic preparation method according to claim 1 is characterized in that the aqueous phase solution is selected from water and/or buffer solution.
  7. According to the microfluidic preparation method of claim 6, the buffer solution is selected from one or more of phosphate buffer, tris(hydroxymethyl)aminomethane buffer, 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid buffer and 0.9% NaCl solution.
  8. According to the microfluidic preparation method of claim 1, the total flow rate of the lipid phase solution and the aqueous phase solution in step S2) is 10.5-30 mL/min; The flow rate ratio of the lipid phase solution to the aqueous phase solution is 1:1 to 1:20.
  9. According to the microfluidic preparation method of claim 1, the total flow rate of the lipid phase solution and the aqueous phase solution in step S2) is 12-24 mL/min; The flow rate ratio of the lipid phase solution to the aqueous phase solution is 1:5.
  10. According to the microfluidic preparation method of claim 1, the lipid phase solution and aqueous phase solution in step S2) are respectively introduced into the microfluidic chip through an injection pump or a constant flow pump.
  11. According to claim 1, the microfluidic preparation method is characterized in that the lipid phase solution and the aqueous phase solution are respectively introduced into the microfluidic chip and the fat body solution is collected, and the fat body solution is concentrated to obtain a fat body delivery platform.
  12. According to the microfluidic preparation method of claim 11, the concentration is ultrafiltration concentration; the molecular weight cutoff of the ultrafiltration concentration is 10-100 kDa.
  13. The fat body delivery platform prepared by the microfluidic preparation method according to any one of claims 1 to 12.
  14. The fat body delivery platform according to claim 13 is characterized in that the average size of the fat body delivery platform is 50-200 nm.
  15. The use of the fat body delivery platform of claim 13 in the preparation of drugs or vaccines for the prevention and treatment of diseases.
  16. The application according to claim 15 is characterized in that the disease is cancer.
  17. A drug or vaccine, characterized in that the drug or vaccine comprises the fat body delivery platform of claim 13 and pharmaceutically acceptable excipients.

Description

A microfluidic preparation method for a fat body delivery platform This application claims priority to Chinese Patent Application No. 202411523787.X, filed on October 29, 2024, entitled “A Microfluidic Preparation Method for a Fat Body Delivery Platform”, the entire contents of which are incorporated herein by reference. Technical Field This invention belongs to the field of biomedical technology, and in particular relates to a microfluidic preparation method for a fat body delivery platform. Background Technology Liposomes are artificially synthesized nanospheres that mimic the lipid structure of lipoproteins or lipid droplets in cells. The main structural feature of liposomes is a neutral lipid core encased in a monolayer of phospholipid molecules, with a hydrophobic interior and a hydrophilic exterior (30–200 nm). Liposomes differ significantly in structure from previously disclosed liposome nanoparticles, lipid nanoparticles delivering nucleic acids, and exosome nanoparticles. The internal hydrophobic neutral lipid core gives liposomes a natural advantage in delivering hydrophobic small molecules, while the specific peripheral monolayer phospholipid membrane mimics the phospholipid membrane structure of lipid droplets or lipoproteins, enabling the recruitment of specific proteins and providing a basis for designing targeted molecules. The main methods for preparing nanoscale emulsions include high-pressure homogenization, microfluidics, ultrasonic methods, phase-inversion emulsification, and self-emulsification. The basic principle of high-pressure homogenization is to utilize the high pressure (up to 300 MPa) generated in the chamber during the delivery stroke, forcing the coarse emulsion through micron-sized orifices in the homogenizer valve. Turbulence, shear stress, and cavitation transform the coarse emulsion into finer droplets. The high-pressure homogenization emulsification process mainly consists of two stages: first, the dispersed phase is disrupted, leading to the formation of tiny droplets in the homogenization chamber, increasing the surface area; then, emulsifier molecules accumulate at the newly formed interface, stabilizing the droplets. Repeated high-pressure homogenization can generate numerous nanodroplets. In the microfluidics method, lipid raw materials are fed into a constant-pressure pump system and then pushed through reaction channels with a fixed geometry at pressures up to 30,000 psi (2,068 bar). The lipid raw materials undergo consistent high shear rates and impact forces within the channels, allowing them to be emulsified into micro/nano emulsions. Compared to traditional high-pressure homogenization methods, microfluidics can produce micro/nanoemulsions with smaller particle sizes and better uniformity in particle size distribution. Ultrasonic methods utilize the mechanical vibrations and ultrasonic cavitation generated by ultrasound to prepare nanoemulsions. The strong shock waves generated by sound waves and cavitation create high pressure and turbulence, causing microdroplets to collapse and break up, producing fine nanoemulsions. However, ultrasonic treatment requires optimal input energy to achieve the smallest droplet diameter; too high or too low energy will lead to poor emulsification. Ultrasonic probes are commonly used to prepare nanoemulsions by emulsifying droplets, and benchtop ultrasonic instruments are also used for small-scale nanoemulsion production. The above are commonly used high-energy input methods for preparing nanoemulsions. Some low-energy input methods are also used to prepare nanoemulsions. Phase transfer emulsification is an emulsification method designed based on the property that some emulsifiers change their hydrophilicity or lipophilicity with temperature under a fixed composition. Some nonionic surfactants change their spontaneous curvature due to hydration of their polar ends, forming oil-in-water emulsions at low temperatures and decreasing their curvature at high temperatures as the solubility of the emulsifier in water increases, thus forming water-in-oil emulsions. The temperature at which oil-in-water emulsions transform into water-in-oil emulsions is called the phase transition temperature. Emulsions can form near the phase transition temperature, but these are unstable and require rapid cooling or heating to generate kinetically stable emulsions with small particle sizes and narrow particle size distributions. Self-emulsification is also a low-energy-input method for emulsion synthesis. This method involves mixing water, oil, and emulsifier at a specific temperature, and then using gentle mechanical stirring to allow the emulsifier to enter the aqueous phase, leading to an increase in the oil-water interfacial area and the formation of nanodroplets. However, the emulsion synthesis methods described above each have their disadvantages. High-pressure homogenization easily forms amorphous membrane structures mixed in the emulsion; phase