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CN-117718016-B - Blood perfusion adsorbent, preparation method and application thereof

CN117718016BCN 117718016 BCN117718016 BCN 117718016BCN-117718016-B

Abstract

The invention belongs to the technical field of biological medicines, and relates to a blood perfusion adsorbent, a preparation method and application thereof. The blood perfusion adsorbent is of a microsphere structure, the carrier of the microsphere is chitosan microsphere, the ligand is I-type collagen, and the particle size of the microsphere is 250-1000 mu m. The blood perfusion adsorbent provided by the invention has the characteristics of large bilirubin adsorption amount, high selective adsorptivity, small albumin adsorption and the like based on the synergistic effect of collagen and chitosan, and has the advantages of excellent bilirubin specific adsorption effect, low cost and good clinical application prospect.

Inventors

  • WANG SHENQI
  • ZHOU LEI
  • ZHOU WAN

Assignees

  • 华中科技大学

Dates

Publication Date
20260508
Application Date
20231229

Claims (12)

  1. 1. The hemoperfusion adsorbent is characterized in that the adsorbent has a microsphere structure, the carrier of the microsphere is chitosan microsphere, the ligand is type I collagen, the particle size of the microsphere is 250-1000 mu m, and the microsphere has the following chemical structure: Wherein, the Represents chitosan microsphere, X represents polyamine.
  2. 2. The hemoperfusion adsorbent of claim 1, wherein the chitosan microsphere is prepared by a reversed phase suspension polymerization method, the chitosan microsphere is used as a carrier structure after freeze drying and pore making, the activated chitosan microsphere carrier is activated by epoxy chloropropane, polyamine is added for reaction, water is used for washing to remove excessive polyamine, the residual polyamine is transferred to an organic phase containing carbonyl diimidazole for reaction, after the reaction is finished, the microsphere is washed by an organic phase, and then the collagen grafting modified chitosan microsphere is obtained by reaction with a collagen solution.
  3. 3. The hemoperfusion adsorbent of claim 1, wherein the surface porosity of the hemoperfusion adsorbent is 35-50% and the collagen loading on the surface of the chitosan microsphere is 0.5-10 mg/g.
  4. 4. The hemoperfusion adsorbent of claim 1, wherein the polyamine is diamine substance, diethylenetriamine or triethylenetetramine with carbon chain length of 2-6, and the collagen loading amount on the surface of the chitosan microsphere is 2-8 mg/g.
  5. 5. The hemoperfusion adsorbent of claim 1 wherein the type I collagen has a molecular weight of 20 kDa-1000 kDa.
  6. 6. The hemoperfusion adsorbent of claim 1, wherein the collagen graft modified chitosan microsphere has a bilirubin clearance of 75-95% and albumin clearance of less than 3%.
  7. 7. The method for preparing the hemoperfusion adsorbent according to claim 1, comprising the steps of: (1) Preparing chitosan microspheres by an inverse suspension polymerization method; (2) Freeze drying chitosan microsphere to prepare holes; (3) The chitosan microsphere obtained in the step (2) is subjected to collagen modification, wherein the collagen modification method comprises the steps of activating the chitosan microsphere through epoxy chloropropane to obtain an activated chitosan microsphere carrier, adding polyamine for reaction, washing with water to remove redundant polyamine, transferring the polyamine to an organic phase containing carbonyl diimidazole for reaction, washing the microsphere with the organic phase after the reaction is completed, and then reacting with a collagen solution; (4) And after the reaction is finished, carrying out post-treatment to obtain the collagen grafting modified chitosan microsphere, namely the blood perfusion adsorbent.
  8. 8. The method of claim 7, wherein the step (1) comprises dissolving chitosan powder in acetic acid to obtain chitosan/acetic acid solution with a mass fraction of 0.5% -5%, pouring the chitosan/acetic acid solution into the dispersed oil phase, wherein the volume ratio of the chitosan acetic acid solution to the oil phase is 1:5-20, dispersing into uniform droplets by adjusting the rotation speed, adding a cross-linking agent, cross-linking for a period of time, collecting microspheres, and processing to obtain the chitosan microspheres.
  9. 9. The method of manufacturing according to claim 8, wherein: The rotating speed of the step (1) is 200-500 rpm, the cross-linking agent of the step (1) is formaldehyde, glutaraldehyde, genipin or tripolyphosphate, the reaction temperature is 40-60 ℃ after the cross-linking agent is added, and the reaction time is 2-6 h.
  10. 10. The method of manufacturing according to claim 7, wherein: The molecular weight of the type I collagen is 20 kDa-1000 kDa, and the mass fraction of the collagen solution is 0.01-2%; When the chitosan microsphere is frozen and dried to prepare holes, the freezing temperature is-80 to-20 ℃ and the time is 12-48 h; reacting the chitosan microsphere carrier activated in the step (3) with a collagen solution for 12-36 h; the polyamine in the step (3) is diamine substance with carbon chain length of 2-6, diethylenetriamine or triethylenetetramine, the reaction after adding polyamine is 2-6 h at 40-60 ℃, and the reaction after adding carbonyl diimidazole is 6-24 h at room temperature.
  11. 11. Use of the hemodynamic adsorbent of claim 1 as a hemodynamic filler for preparing an adsorption column in a hemodynamic filler.
  12. 12. A hemodiafiltration device obtained by using the hemodiafiltration adsorbent of claim 1 as a hemodiafiltration device filler.

Description

Blood perfusion adsorbent, preparation method and application thereof Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to a blood perfusion adsorbent, a preparation method and application thereof. Background Bilirubin is one of the major products of hemoglobin metabolism and is a pathogenic toxin. It binds to albumin and is transported to the liver for final excretion. In the case of patients with liver dysfunction, the metabolic pathway is blocked, and bilirubin cannot be discharged out of the body in time, so that hyperbilirubinemia is caused. Excessive bilirubin can accumulate in various tissues such as the brain, eventually leading to brain damage and even death. Hyperbilirubinemia is generally defined as total bilirubin >12mg/dL, whereas normal serum bilirubin levels are 0.4-1.8mg/dL, and therefore, it is critical to remove excess bilirubin from the blood of hyperbilirubinemia patients. To address this problem, various techniques such as hemodialysis, phototherapy, plasma exchange, and blood perfusion are used to remove excess bilirubin. Among these techniques, blood perfusion has proven to be one of the most effective methods for bilirubin removal (J. Mater. Chem. B,2017,5 (29): 5763-5773), the core functional unit of which is an adsorbent material. Currently, various bilirubin adsorbents such as activated carbon, chitosan or polystyrene and other high molecular materials and novel materials such as MoFs or PAF have been widely developed. However, these adsorbents exhibit limitations in terms of bilirubin adsorption performance or blood compatibility, limiting their clinical use (ACS APPLIED MATERIALS & Interfaces,2020 (12): 25546-25556). Therefore, developing a new bilirubin adsorbent is still urgent, and designing a bilirubin adsorbent with high adsorption performance, good blood compatibility, and low cost is not only a great demand for blood perfusion application, but also a great challenge. Chitosan is a natural high molecular polysaccharide that is produced by deacetylation of chitin. Because of its good biocompatibility and hemocompatibility, it is widely used in the adsorbent field. However, chitosan itself has limited adsorption capacity for free bilirubin and bilirubin in albumin-rich solutions and has a high adsorption capacity for albumin (Colloids Surfaces B: biointerfaces,2013,112 (12): 103-107). Chitosan generally enhances the adsorption capacity for bilirubin by complexing with other inorganic substances or grafted ligands (J. Mater. Chem. B,2022, (10): 8650-8663). However, these inorganic materials have high adsorptivity to components such as albumin in blood, and the safety has yet to be further improved and long-term experimental verification has been required (Topics in Current Chemistry,2020,378 (1): 1-41). Bilirubin has the characteristics of negative charge and strong hydrophobicity, and common ligands typically include polylysine, cyclodextrin, quaternary amine groups, and the like (ARTIFICIAL ORGANS,1992,16 (6): 568-576;Journal of Applied Polymer Science,2013,130 (1): 563-571). However, these ligands tend to have limited adsorption properties for bilirubin, or have higher adsorption for other components in the blood such as albumin, etc., resulting in poor blood compatibility. Disclosure of Invention The invention aims at overcoming the defects of the prior art and provides a blood perfusion adsorbent, a preparation method and application. The blood perfusion adsorbent provided by the invention has the characteristics of large bilirubin adsorption amount, high selective adsorptivity, small albumin adsorption and the like based on the synergistic effect of collagen and chitosan, and has an excellent bilirubin specific adsorption effect. In order to achieve the above purpose, the invention adopts the following technical scheme: The first aspect of the present invention provides a hemoperfusion adsorbent which is in a microsphere structure, wherein the carrier of the microsphere is chitosan microsphere, the ligand is type I collagen, the particle size of the microsphere is 250-1000 μm, and the microsphere has the following chemical structure: Wherein, the Represents chitosan microsphere, X represents polyamine. According to the scheme, the hemoperfusion adsorbent takes chitosan microspheres as a carrier after freeze drying and pore making. According to the scheme, the blood perfusion adsorbent is prepared by adopting a reverse-phase suspension polymerization method to prepare chitosan microspheres, wherein the chitosan microspheres are used as a carrier structure after freeze-drying and pore-forming, and react with collagen, or react with collagen after being activated by epoxy chloropropane, and are obtained by taking polyamine as a spacer arm. According to the scheme, the surface of the blood perfusion adsorbent microsphere is provided with gaps, preferably, the surface porosity of the blood perfusion adsorbent is 3