Polymeric microbeads and method of preparation

• US 5,653,922 A
• Filed: 06/07/1995
• Issued: 08/05/1997
• Est. Priority Date: 06/06/1994
• Status: Expired due to Term

• Current Assignee: Sunstorm Research Corporation
• Original Assignee: Biopore Corporation
• Inventors: Benson, James R., Kitagawa, Naotaka, Li, Nai-Hong
• Primary Examiner(s): Nutter, Nathan M.
• Application Number:
  US08/485,494
• Time in Patent Office: 790 Days
• Field of Search: 264/4.3, 264/4.33, 264/4.7
• US Class Current: 264/4.3
• CPC Class Codes: B01D 15/1807 using counter-currents, e.g. fluidised beds
• CPC Class Codes: B01D 15/362 Cation-exchange
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01J 13/02 Making microcapsules or microballoons for medical preparations A61K9/50
• CPC Class Codes: B01J 13/18 In situ polymerisation with all reactants being present in the same phase
• CPC Class Codes: B01J 13/185 in an organic phase
• CPC Class Codes: B01J 13/20 After-treatment of capsule walls, e.g. hardening
• CPC Class Codes: B01J 20/267 Cross-linked polymers
• CPC Class Codes: B01J 20/28004 Sorbent size or size distribution, e.g. particle size
• CPC Class Codes: B01J 20/28011 Other properties, e.g. density, crush strength
• CPC Class Codes: B01J 20/28019 Spherical, ellipsoidal or cylindrical
• CPC Class Codes: B01J 20/28069 Pore volume, e.g. total pore volume, mesopore volume, micropore volume
• CPC Class Codes: B01J 20/28085 being more than 50 nm, i.e. macropores
• CPC Class Codes: B01J 20/2809 Monomodal or narrow distribution, uniform pores
• CPC Class Codes: B01J 20/285 based on polymers
• CPC Class Codes: B01J 20/288 Polar phases
• CPC Class Codes: B01J 20/3064 Addition of pore forming agents, e.g. pore inducing or porogenic agents
• CPC Class Codes: B01J 20/321 consisting of a polymer obtained by reactions involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3251 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
• CPC Class Codes: B01J 2220/54 Sorbents specially adapted for analytical or investigative chromatography
• CPC Class Codes: B01J 39/26 : Cation exchangers for chromatographic processes
• CPC Class Codes: B01J 41/20 : Anion exchangers for chromatographic processes
• CPC Class Codes: C07K 1/042 : characterised by the nature of the carrier
• CPC Class Codes: C08F 212/08 : Styrene
• CPC Class Codes: C08F 2800/10 : as molar percentages
• CPC Class Codes: C08F 8/00 : Chemical modification by after-treatment graft polymers, block polymers, crosslinking with unsaturated monomers or with polymers C08F251/00 - C08F299/00; of conjugated diene rubbers C08C
• CPC Class Codes: C08F 8/24 : Haloalkylation
• CPC Class Codes: C08F 8/30 : Introducing nitrogen atoms or nitrogen-containing groups
• CPC Class Codes: C08F 8/36 : Sulfonation; Sulfation
• CPC Class Codes: C08F 8/44 : Preparation of metal salts or ammonium salts
• CPC Class Codes: C12N 2533/30 : Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0075 : using microcarriers
• CPC Class Codes: Y10S 521/902 : Cellular polymer containing an isocyanurate structure

Abstract

The present invention relates to porous crosslinked polymeric microbeads having cavities joined by interconnecting pores wherein at least some of the cavities at the interior of each microbead communicate with the surface of the microbead. The present invention also relates to a process for producing a porous, crosslinked polymeric microbead as well as the product of this process. This process involves combining an oil phase with an aqueous discontinuous phase to form an emulsion, adding the emulsion to an aqueous suspension medium to form an oil-in-water suspension of dispersed emulsion droplets, and polymerizing the emulsion droplets to form microbeads. At least 10% of the microbeads produced in accordance with the present invention are substantially spherical or substantially ellipsoidal or a combination of the two.

Claims

1. Porous crosslinked polymeric microbeads having cavities joined by interconnecting pores wherein at least some of the cavities at the interior of each microbead communicate with the surface of the microbead and wherein approximately 10% of the microbeads are substantially spherical or substantially ellipsoidal or a combination thereof.

2. The microbeads of claim 1 wherein the microbeads have an average diameter in the range of about 5 μ

m to about 5 mm.

3. The microbeads of claim 1 wherein the microbeads have a void volume of at least about 70%.

27. The microbeads of claim 3 wherein approximately 20% of the microbeads are substantially spherical or substantially ellipsoidal or a combination thereof.

28. The microbeads of claim 27 wherein approximately 50% of the microbeads are substantially spherical or substantially ellipsoidal or a combination thereof.

29. The microbeads of claim 3 wherein the microbeads have an average diameter in the range of about 50 μ

m to about 500 μ

m.

30. The microbeads of claim 3 wherein the bulk density of the microbeads is in the range of about 0.05 gm/ml to about 0.2 gm/ml.

31. The microbeads of claim 3 wherein the microbeads have been functionalized such that the functionalized microbeads absorb aqueous and/or organic acidic liquids, said functionalized microbeads comprising the structural unit:

##STR8## in which A is a crosslinked carbon chain;

Y is an optional spacer group; and

Z is an ionic or polar functional group.

32. The functionalized microbeads of claim 31 wherein the ratio of aqueous and/or organic acid absorption by the functionalized microbeads to methyl oleate absorption is greater than about 1.2.

33. The functionalized microbeads of claim 31 wherein Z is selected from the group consisting of

an amino or substituted amino group;

an alkyl cationic quaternary ammonium group having 8 or more carbon atoms; and

an alkyl cationic quaternary ammonium group of 8 carbon atoms or less in the presence of a counterion that is an organic group containing 8 or more carbon atoms.

34. The functionalized microbeads of claim 31 wherein Z comprises an ionic or polar functional group selected from the group consisting of species 1-3:

##STR9## in which R₂, R₃, R₄, R₅, and R₆ can be the same or different and are selected from the group consisting of an alkyl, cycloalkyl, aryl, hydroxyalkyl;

or R₂ and R₃ form part of a ring system; and

the number of carbons present in R₄ +R₅ +R₆ is 10 or more for a cationic quaternary ammonium (3);

the number of carbons present in R₄ +R₅ is 8 or more for an amine salt (2); and

the counterion X^- is an organic or inorganic ion.

35. The functionalized microbeads of claim 34 wherein

the number of carbons present in R₄ +R₅ +R₆ is less than 10 for a cationic quaternary ammonium (3);

the number of carbons present in R₄ +R₅ is less than 8 for an amine salt (2); and

the counterion X^- is an organic group having 8 or more carbons.

36. The functionalized microbeads of claim 31 wherein the level of crosslinking is from about 1 to about 20%.

37. The functionalized microbeads of claim 31 wherein the level of crosslinking is from about 2 to about 10%.

38. The functionalized microbeads of claim 31 wherein the void volume of the microbeads is greater than about 80%.

39. The functionalized microbeads of claim 31 wherein the degree of functionalization is greater than about 50%.

40. The microbeads of claim 3 wherein the microbeads have been functionalized such that the functionalized microbeads absorb aqueous liquids, said functionalized microbeads comprising the structural unit:

##STR10## in which A is a crosslinked carbon chain;

Y is an optional spacer group; and

Z is an ionic or polar functional group.

41. The functionalized microbeads of claim 40 wherein the ratio of 10% sodium chloride absorption by the functionalized microbeads to water absorption is greater than about 0.1.

42. The functionalized microbeads of claim 41 wherein the ratio of 10% sodium chloride absorption to water absorption is greater than about 0.5.

43. The functionalized microbeads of claim 42 wherein the ratio of 10% sodium chloride absorption to water absorption is greater than about 0.7.

44. The functionalized microbeads of claim 40 wherein Z is selected from the group consisting of

an alkyl cationic quaternary ammonium group having 10 carbon atoms or less;

an alkyl amine salt group having 8 carbon atoms or less;

an alkoxylate group;

a metal, ammonium, or substituted ammonium salt of a sulfuric, carboxylic, phosphoric or sulfonic acid group, except that where Z is a sulfonic acid;

Y does not have the structure;

##STR11##

45. The functionalized microbeads of claim 40 wherein Z comprises an ionic or polar functional group selected from the group consisting of structures 1-2:

##STR12## in which R₂, R₃, and R₄ can be the same or different and are selected from the group consisting of an alkyl, cycloalkyl, aryl, and hydroxyalkyl;

or R₂ and R₃ form part of a ring system; and

the number of carbons present in R₂ +R₃ +R₄ is less than 10 for a cationic quaternary ammonium (1);

the number of carbons present in R₂ +R₃ is less than 8 for an amine salt (2); and

the counterion X^- is an inorganic ion or a carboxylate group having less than 8 carbons.

46. The functionalized microbeads of claim 40 wherein Z is an alkoxylate group having the structure:

##STR13## where p is 1 to 680, and ##STR14## and where R₅ is a hydrogen or an alkyl group, and M is a metal, an ammonium, or a substituted ammonium cation.

47. The functionalized microbeads of claim 40 wherein the level of crosslinking is from about 1 to about 20%.

48. The functionalized microbeads of claim 40 wherein the level of crosslinking is from about 2 to about 10%.

49. The functionalized microbeads of claim 40 wherein the void volume of the microbeads is greater than about 80%.

50. The functionalized microbeads of claim 40 wherein the degree of functionalization is greater than about 50%.

51. The microbeads of claim 3 wherein the microbeads have a carboniferous structure.

52. The carboniferous microbeads of claim 51 wherein the microbeads have a void volume of at least about 80%.

53. The microbeads of claim 3 additionally comprising a gel or pre-gel within the cavities of the microbeads, wherein the gel or pre-gel comprises a chemical group that interacts with a chemical entity selected from the group consisting of a component of a solution and a reactant in a chemical synthesis.

54. The microbeads of claim 53 wherein the cavity size is in the range of about 1 to about 50 μ

m in diameter.

55. The microbeads of claim 53 wherein the microbeads are crosslinked such that, in the presence of liquid, microbead swelling does not exceed approximately two times the microbead dry volume.

56. The microbeads of claim 53 wherein the ratio by weight of swollen gel to microbeads lies in the range of about 60:

40 to about 95;

5.

57. A process for producing the microbeads of claim 53 comprising

a. producing porous, crosslinked polymeric microbeads having cavities joined by interconnecting pores and

b. depositing and retaining a gel or pre-gel in the microbead cavities.

58. The process of claim 57 wherein step (b) comprises contacting the microbeads with a solution comprising pre-gel components and a swelling solvent for the microbeads for a time sufficient to allow the pre-gel components to permeate the swollen microbeads and form a gel within the microbead cavities.

59. The microbeads of claim 3 wherein a molecule selected from the group consisting of a polypeptide and an oligonucleotide is attached to the surface of the microbeads.

60. The microbeads of claim 59 wherein the molecule is a polypeptide selected from the group consisting of an antibody, a lectin, an enzyme, and a hapten.

61. The microbeads of claim 59 wherein the molecule is attached to the surface of the microbeads by a type of attachment selected from the group consisting of non-covalent adsorption, covalent bonding, and attachment via a bridging molecule.

4. A process for producing a porous, crosslinked polymeric microbead comprising

(a) combining

(i) an oil phase comprising

(1) a substantially water-insoluble, monofunctional monomer;

(2) a substantially water-insoluble, polyfunctional crosslinking agent;

(3) an emulsifier that is suitable for forming a stable water-in-oil emulsion; and

(4) a stabilizer, wherein said stabilizer is a film-forming compound that is soluble in an organic solvent, and said stabilizer is present in the oil phase at a concentration in the range of about 0.01 to about 15 weight percent; and

(ii) an aqueous discontinuous phase to form an emulsion, wherein the emulsion comprises at least about 70% aqueous discontinuous phase;

(b) adding the emulsion to an aqueous suspension medium to form an oil-in-water suspension of dispersed emulsion droplets; and

(c) polymerizing the emulsion droplets.

5. The process of claim 4 wherein the monomer comprises a styrene-based monomer.

6. The process of claim 4 wherein the monomer is present in the oil phase at a concentration in the range of about 4 to about 90 weight percent.

7. The process of claim 4 wherein the crosslinking agent comprises an agent selected from the group consisting of a divinylbenzenic compound, a di- or triacrylic compound, and a triallyl isocyanurate.

8. The process of claim 4 wherein the crosslinking agent is present in the oil phase at a concentration in the range of about 1 to about 89 weight percent.

9. The process of claim 4 wherein the emulsifier comprises an agent selected from the group consisting of a sorbitan fatty acid ester, a polyglycerol fatty acid ester, and a polyoxyethylene fatty acid or ester.

10. The process of claim 4 wherein the emulsifier is present in the oil phase at a concentration in the range of about 3 to about 50 weight percent.

11. The process of claim 4 wherein the oil-in-water suspension comprises an amount of high internal phase emulsion suitable for generating a stable suspension.

12. The process of claim 4 wherein the aqueous suspension medium comprises a suspending agent.

13. The process of claim 12 wherein the suspending agent comprises an agent selected from the group consisting of a water-soluble polymer and a water-insoluble inorganic solid.

14. The process of claim 12 wherein the suspending agent comprises a natural gum.

15. The process of claim 12 wherein the suspending agent is present in the aqueous suspension medium at a concentration of about 1 to about 30 weight percent.

16. The process of claim 4 wherein the suspension is formed by adding the high internal phase emulsion to the aqueous suspension medium while providing sufficient shear agitation to generate a stable suspension.

17. The process of claim 4 wherein the oil phase additionally comprises an oil-soluble polymerization initiator, and no polymerization initiator is present in either the aqueous discontinuous phase or the aqueous suspension medium.

18. The process of claim 17 wherein the oil-soluble initiator is selected from the group consisting of AIBN, benzoyl peroxide, lauroyl peroxide, and a VAZO initiator.

19. The process of claim 17 wherein the oil-soluble polymerization initiator is present in the oil phase at a concentration of up to about 5 weight percent of total polymerizable monomer.

20. The process of claim 17 wherein the aqueous discontinuous phase consists essentially of water.

21. The process of claim 17 wherein the stabilizer comprises a natural or synthetic polymeric stabilizer.

22. The process of claim 21 wherein the polymeric stabilizer comprises a cellulose derivative.

23. The process of claim 21 wherein the polymeric stabilizer comprises an agent selected from the group consisting of methyl cellulose, ethyl cellulose, and partially hydrolyzed poly(vinyl alcohol).

26. The process of claim 17 wherein the oil phase additionally comprises a porogen.

62. The process of claim 23, wherein the polymeric stabilizer comprises ethyl cellulose.

63. The process of claim 4 wherein the stabilizer is present in the oil phase at a concentration in the range of about 0.01 to about 1 weight percent.

64. The process of claim 63 wherein the stabilizer is present in the oil phase at a concentration in the range of about 0.02 to about 0.6 weight percent.

65. The process of claim 21 wherein the emulsion additionally comprises an inert solvent that is capable of solubilizing the stabilizer and is miscible in the oil phase of the HIPE.

66. The process of claim 65 wherein the inert solvent is a halogenated solvent.

67. The process of claim 65 wherein the inert solvent is selected from the group consisting of trichloroethane, toluene, and chloroform.

68. The process of claim 65 wherein the inert solvent is present in the oil phase at a concentration in the range of about 3 to about 60 weight percent.

69. The process of claim 68 wherein the inert solvent is present in the oil phase at a concentration in the range of about 10 to about 40 weight percent.

70. The process of claim 14 wherein the suspending agent comprises acacia gum.

71. The process of claim 15 wherein the suspending agent is present in the aqueous suspension medium at a concentration of about 2 to about 15 weight percent.

24. A process for producing a porous crosslinked polymeric microbead comprising

(a) combining

(i) an oil phase comprising

(1) styrene;

(2) divinylbenzene;

(3) lauroyl peroxide;

(4) trichloroethane;

(5) sorbitan monooleate; and

(6) ethylcellulose; and

(ii) an aqueous discontinuous phase that does not contain a polymerization initiator to form an emulsion;

(b) adding the emulsion to an aqueous suspension medium to form an oil-in-water suspension of dispersed emulsion droplets, wherein said suspension medium comprises acacia gum and does not contain a polymerization initiator; and

(c) polymerizing the emulsion droplets.

25. A porous crosslinked polymeric microbead produced by the process comprising

(a) combining

(i) an oil phase comprising

(1) styrene;

(2) divinylbenzene;

(3) lauroyl peroxide;

(4) trichloroethane;

(5) sorbitan monooleate; and

(6) ethylcellulose; and

(ii) an aqueous discontinuous phase that does not contain a polymerization initiator to form an emulsion;

(b) adding the emulsion to an aqueous suspension medium to form an oil-in-water suspension of dispersed emulsion droplets, wherein said suspension medium comprises acacia gum and does not contain a polymerization initiator; and

(c) polymerizing the emulsion droplets.