Method for the preparation of microspheres which contain colloidal systems

• US 6,395,302 B1
• Filed: 02/15/2000
• Issued: 05/28/2002
• Est. Priority Date: 11/19/1996
• Status: Expired due to Fees

• Current Assignee: Octoplus BV
• Original Assignee: Octoplus BV
• Inventors: Hennink, Wilhelmus Everhardus, Franssen, Okke
• Primary Examiner(s): Page, Thurman K.
• Assistant Examiner(s): TRAN, SUSAN T
• Application Number:
  US09/503,847
• Time in Patent Office: 833 Days
• Field of Search: 424/490,489,400,497,417 264/4.3,4.1 427/213,36
• US Class Current: 424/489
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1652 Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin homeopathic globules A61K9/1623
• CPC Class Codes: A61K 9/1664 Compounds of unknown constitution, e.g. material from plants or animals oils, fats, waxes, shellac A61K9/1617
• CPC Class Codes: A61K 9/1694 resulting in granules or microspheres of the matrix type containing more than 5% of excipient
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: Y10S 977/906 Drug delivery

Abstract

The invention relates to a method for the preparation of microencapsulated colloidal systems such as liposomes, i.e., microspheres which comprise colloidal systems. These microencapsulated colloidal systems can be used as controlled release systems for the delivery of active ingredients in in vivo and in vitro applications. A method is provided in which the colloidal systems are added to a phase which comprises a water soluble crosslinkable polymer followed by formation of microspheres.

Claims

1. A method for preparing microspheres encapsulating colloidal systems, which method comprises the steps of:

a) providing an aqueous mixture of

i) a first phase comprising a first water soluble crosslinkable polymer,

ii) a second phase comprising a second water soluble polymer which is incompatible in solution with the polymer in said first phase, and

iii) colloidal systems to be suspended in said first phase;

b) forming an emulsion of said first phase in said second phase;

c) forming microspheres in the emulsion by crosslinking at least part of said crosslinkable polymer, thus forming said microspheres encapsulating colloidal systems; and

d) recovering the microspheres;

wherein said colloidal systems are selected from the group consisting of liposomes, iscoms, polyplexes, lipoplexes, nanoparticles, solid lipid particles in the colloidal size range, emulsions, and combinations thereof.

2. The method of claim 1, in which step a) is carried out by forming a premixture by contacting said colloidal systems with said first phase, and subsequently contacting this premixture with said second phase.

3. The method of claim 1, in which said crosslinkable polymer is a dextran polymer which comprises crosslinkable groups.

4. The method of claim 3, in which said crosslinkable groups are methacrylate groups.

5. The method of claim 4, in which said dextran polymer is selected from the group consisting of dexMA, dexHEMA and dexLactateHEMA.

6. The method of claim 5, in which the degree of substitution of said dextran polymer is from 2 to 30.

7. The method of claim 1, in which said polymer in the second phase is selected from the group consisting of polyethylene glycol (PEG) and polyvinyl alcohol (PVA).

8. The method of claim 1, wherein said colloidal systems comprise an active ingredient.

12. The method of claim 8, wherein said active ingredient is a protein drug.

15. Microspheres prepared by the method of claim 1.

16. The method of claim 1, wherein said microspheres effect a pulsed release profile of the colloidal systems with a desired lag time.

17. The method of claim 1, wherein said emulsion is a ternary two phase system of

(1) said first water-soluble polymer which is crosslinkable;

(2) said second water-soluble polymer, and

(3) water,

wherein said ternary system is characterized by a phase diagram and wherein said ternary system forms one phase when composition of the ternary system is below the binodal in said phase diagram and forms two phases when the composition of the ternary system is above the binodal in said phase diagram, and wherein the composition of said ternary two phase system provided is above said binodal and is capable of forming a continuous phase and a discontinuous phase;

and wherein said emulsion comprises a continuous and discontinuous phase, wherein said first, crosslinkable, polymer is the discontinuous phase in said two-phase system and the second polymer is the continuous phase in said two-phase system, said two phase system having been obtained by adjusting the amounts of water, first polymer and second polymer so as to place the system above the binodal, and

wherein said colloidal system is present in said two-phase system.

18. The method of claim 1, wherein the degree of crosslinking and water content are adjusted so as to result in pore sizes in said microspheres which are equal to or less than the hydrodynamic diameter of the colloidal system, said pores thus entrapping said colloidal system.

19. The method of claim 1, wherein said first crosslinkable polymer, when crosslinked, can be degraded by virtue of comprising bonds that are hydrolyzable under physiological conditions, and said degradation of the crosslinked polymer occurs through hydrolyzing said bonds.

20. The method of claim 1, wherein said first crosslinkable polymer, when crosslinked, can be degraded by virtue of including, in said microspheres, at least one enzyme which degrades said polymer, and wherein said degradation of crosslinked polymer occurs through action of said enzyme on said polymer.

21. The method of claim 20, wherein said first crosslinkable polymer is dextran or derivatized dextran and the enzyme which degrades the polymer is dextranase.

22. The method of claim 1, wherein said crosslinking is formed by crosslinking methacrylate or a methacrylate derivative or by using isocyanate or through poly-N-isopropyl acrylamide.

23. The method of claim 1, wherein step (d) is performed by filtration or centrifugation.

9. Microspheres, at least 80 wt. % thereof having a particle diameter of between 100 nm and 100 μ

m, which microspheres are comprised of a degradable crosslinked polymer which encapsulates colloidal systems wherein said colloidal systems are selected from the group consisting of liposomes, iscoms, polyplexes, lipoplexes, nanoparticles, solid lipid particles in the colloidal size range, emulsions, and combinations thereof.

10. Microspheres according to claim 9, being free from organic solvent.

11. Microspheres according to claim 9, wherein said colloidal systems comprise an active ingredient.

13. The microspheres of claim 9, wherein at least 80% of said microspheres have a particle diameter of between 5 and 15 μ

m.

14. The microspheres of claim 11, wherein said active ingredient is a protein drug.

24. The microspheres of claim 9, wherein said microspheres effect a pulsed release profile of the colloidal system with a desired lag time.

25. The microspheres of claim 9, wherein the pore size of the crosslinked polymer is equal to or less than the hydrodynamic diameter of the colloidal system.

26. The microspheres of claim 9, wherein said degradable, crosslinked polymer is selected from the group consisting of dextran, derivatized dextran, starch, starch derivatives, cellulose derivatives, proteins and derivatized proteins.

27. The microspheres of claim 26, wherein said degradable, crosslinked polymer is dextran or a derivatized dextran.

28. The microspheres of claim 27, wherein said derivatized dextran comprises methacrylate groups.

29. The microspheres of claim 28, wherein said derivatized dextran is dexMA, dexHEMA or dexLactateHEMA.