Magnetically responsive, hydrophilic microspheres for incorporation of therapeutic substances and methods of preparation thereof

• US 4,871,716 A
• Filed: 12/03/1986
• Issued: 10/03/1989
• Est. Priority Date: 02/04/1986
• Status: Expired due to Fees

• Current Assignee: Eugene P. Goldberg
• Original Assignee: University of Florida (State University System of Florida)
• Inventors: McCluskey, Richard A., Longo, William E., Goldberg, Eugene P.
• Primary Examiner(s): Nucker, Christine M.
• Application Number:
  US06/937,611
• Time in Patent Office: 1,035 Days
• Field of Search: 436/526, 436/528, 436/531, 436/829, 424/9, 424/491, 424/497, 424/499, 424/501, 435/7, 514/2, 514/6, 252/62.51, 252/62.55, 106/124
• US Class Current: 424/9.322
• CPC Class Codes: A61K 9/1641 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
• CPC Class Codes: A61K 9/1658 Proteins, e.g. albumin, gelatin
• CPC Class Codes: A61K 9/1694 resulting in granules or microspheres of the matrix type containing more than 5% of excipient
• CPC Class Codes: A61K 9/5115 Inorganic compounds
• CPC Class Codes: A61K 9/5146 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
• CPC Class Codes: A61K 9/5161 Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
• CPC Class Codes: A61K 9/5169 Proteins, e.g. albumin, gelatin
• CPC Class Codes: A61K 9/5192 Processes
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10S 930/29 Polyamino acid or polypeptide with an uninterrupted series of peptide repeating units
• CPC Class Codes: Y10S 977/788 Of specified organic or carbon-based composition
• CPC Class Codes: Y10S 977/838 Magnetic property of nanomaterial
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

Novel magnetically responsive hydrophilic protein or polypeptide microspheres prepared by dispersing an aqueous solution or dispersion of protein or polypeptide and particulate magnetically responsive material in an organic solvent solution of a high molecular weight polymer to form a stabilized dispersion of microspheres and cross-linking said microspheres with a polyfunctional cross-linking agent.

Claims

1. A method of preparing novel hydrophilic, magnetically responsive microspheres consisting essentially of cross-linked protein or polypeptide particulate and a magnetically responsive material comprising

(a) providing a dispersion of an aqueous solution or dispersion of polypeptide or protein microspheres and a particulate magnetically responsive material in an organic, substantially water immiscible solvent solution of a high molecular weight polymer, said organic solvent being substantially a non-solvent for said microspheres and said polymer solution stabilizing the dispersion of microspheres and magnetically responsive material,

(b) incorporating a polyfunctional cross-linking agent for said protein or polypeptide in said dispersion, and

(c) allowing said cross-linking agent to react with said protein or polypeptide microspheres for a time sufficient to cross-link at least a portion of the microspheres, thereby providing magnetically responsive microspheres containing free reactive functional groups.

2. The method of claim 1 including the step of separating said cross-linked protein or polypeptide microspheres from said dispersion.

3. The method of claim 1 wherein said protein is albumin.

4. The method of claim 3 wherein said albumin is human serum albumin.

5. The method of claim 3 wherein said albumin is bovine serum albumin.

6. The method of claim 1 or 2 wherein said organic solvent solution of high molecular weight polymer is a solution of polymethylmethacrylate in a mixture of toluene and chloroform.

7. The method of claim 1 or 2 wherein said organic solvent solution of high molecular weight polymer is a solution of polyoxyethylene-polyoxypropylene copolymer in chloroform.

8. The method of claim 1 or 2 wherein said organic solvent solution of high molecular weight polymer is a solution of cellulose acetate butyrate in ethylene dichloride.

9. The method of claim 1 or 2 wherein said organic solvent solution of high molecular weight polymer is a solution of bisphenol polycarbonate in chloroform.

10. The method of claim 1 or 2 wherein the concentrations of high molecular weight polymer in organic solvent is from about 1% to about 40%, by weight.

11. The method of claim 1 or 2 wherein said stabilized dispersion of protein or polypeptide microspheres is formed by dispersing an aqueous solution of said protein or polypeptide in said organic solvent solution of high molecular weight polymer.

12. The method of claim 1 or 2 wherein said microspheres have an average size in the range of from about 0.05 microns to about 500 microns.

13. The method of claim 1 or 2 wherein said cross-linking agent is a dialdehyde.

14. The method of claim 13 wherein said cross-linking agent is glutaraldehyde.

15. The method of claim 1 or 2 wherein said cross-linking agent is a diisocyanate.

16. The method of claim 15 wherein said diisocyanate is 2,4-tolylene diisocyanate or 1,6-diisocyanate hexane.

17. The method of claim 1 wherein said magnetically responsive material is magnetite.

18. The product produced according to the method of claim 1 or 2.

19. The product produced according to the method of claim 3.

20. The method of claim 1 including the step of reacting reactive functional groups of said cross-linked microspheres with a first substance containing at least one functional group reactive therewith to form a covalent bond between said cross-linked microspheres and said substance.

21. The method of claim 20 wherein said first substance is a biologically active substance suitable for administration to a biological system.

22. The method of claim 20 wherein said substance contains at least one additional functional group which is non-reactive with the reactive functional groups of said cross-linked microspheres.

23. The method of claim 22 wherein said at least one additional functional group is reacted with a second substance containing a functional group reactive therewith to form a bond therebetween.

24. The method of claim 23 wherein said second substance is a biologically active substance suitable for administration to a biological system.

25. The method of claim 20 wherein said magnetically responsive material is magnetite.

26. The product produced according to the method of claim 20.

27. The product produced according to the method of claim 21.

28. The product produced according to the method of claim 22.

29. The product produced according to the method of claim 23.

30. The product produced according to the method of claim 24.

31. The product produced according to the method of claim 25.

32. A composition in unit dosage form adapted for administration to a biological system comprising a biologically effective amount of the product of any of claims 26 to 31 and a biologically acceptable carrier therefor.

33. A method of administering a biologically active substance to a biological system comprising administering thereto the product of any of claims 26 to 31.