Composite particles, derived conjugates, preparation method and applications

• US 7,217,457 B2
• Filed: 05/06/2002
• Issued: 05/15/2007
• Est. Priority Date: 05/10/2001
• Status: Expired due to Term

• Current Assignee: Bio Merieux, Centre National De La Recherche Scientifique
• Original Assignee: Bio Merieux, Centre National De La Recherche Scientifique
• Inventors: Mandrand, Bernard, Elaissari, Abdelhamid, Pichot, Christian, Montagne, Franck, Bibette, Jeo, Mondain-Monval, Olivier, Bosc, Eric
• Primary Examiner(s): Le; H. Thi
• Application Number:
  US10/475,709
• Publication Number:
  US 20040115433A1
• Time in Patent Office: 1,835 Days
• Field of Search: 428/402, 428/403, 428/407, 427/212, 427/221, 524/401, 524/457, 524/481
• US Class Current: 428/407
• CPC Class Codes: A61K 41/0052 Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
• CPC Class Codes: A61K 49/1854 the organic macromolecular compound being obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly(meth)acrylate, polyacrylamide, polyvinylpyrrolidone, polyvinylalcohol
• CPC Class Codes: A61K 49/1857 the organic macromolecular compound being obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. PLGA
• CPC Class Codes: A61K 49/1887 Agglomerates, clusters, i.e. more than one (super)(para)magnetic microparticle or nanoparticle are aggregated or entrapped in the same maxtrix
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C08F 291/00 Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds according to more than one of the groups C08F251/00 - C08F289/00
• CPC Class Codes: C08F 292/00 Macromolecular compounds obtained by polymerising monomers on to inorganic materials
• CPC Class Codes: C08K 9/08 Ingredients agglomerated by treatment with a binding agent
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

The invention relates to composite particles comprising a hydrophobic polymer core and inorganic nanoparticles. Said hydrophobic polymer forms a polymer matrix inside which the inorganic nanoparticles are stabilized and distributed in a relatively homogenous manner. Said particles are at least partially surrounded by an amphophilic copolymer comprising a hydrophobic part and a hydrophilic part, said hydrophobic part being at least partially immobilized in the polymer matrix. The inventor also relates to the preparation method thereof, the conjugates comprising such particles and a ligand and the use of same in reactants and therapeutic compositions.

Claims

1. A composite particle comprising a core made of a hydrophobic polymer and inorganic nanoparticles, said hydrophobic polymer constituting a polymer matrix within which the inorganic nanoparticles are stabilized and distributed homogeneously, said particle being characterized in that it is at least partially surrounded by an amphiphilic copolymer comprising a hydrophobic part and a hydrophilic part, the hydrophobic part of which is at least partially immobilized on or in said polymer matrix.

2. The particle as claimed in claim 1, characterized in that it is completely surrounded by an amphiphilic copolymer.

3. The particle as claimed in claim 1, characterized in that all of the hydrophobic part of the amphiphilic copolymer is immobilized on or in the polymer matrix.

4. The particle as claimed in claim 1, characterized in that the hydrophobic part of the amphiphilic copolymer is selected from the group consisting of:

polystyrenes, polyalkyls and fatty acid chains.

5. The particle as claimed in claim 1, characterized in that the hydrophilic part of the amphiphilic copolymer is selected from the group consisting of:

poly(acrylic acid)s, polysulfates, polyamines, polyamides and polysaccharides.

6. The particle as claimed in claim 1, characterized in that the amphiphilic copolymer is selected from the group consisting of:

block polymers, sequential polymers, branched polymers and comb polymers.

7. The particle as claimed in claim 6, characterized in that it exhibits reactive functional groups at its surface capable of reacting with at least one ligand.

8. A conjugate, characterized in that it is composed of the composite particle as defined in claim 7, the functional groups of which have interacted, directly or indirectly, with at least one ligand selected from the group consisting of:

antibodies, antibody fragments, proteins, polypeptides, enzymes, polynucleotides, nucleic acid fragments and biotin.

9. A reagent comprising at least one conjugate as defined in claim 8.

10. A conjugate, characterized in that it is composed of the composite particle as defined in claim 7, the functional groups of which have interacted, directly or indirectly, with at least one ligand selected from the group consisting of:

cage molecules, chelating agents and catalysts.

11. A conjugate, characterized in that it is composed of the composite particle as defined in claim 7, the functional groups of which have interacted, directly or indirectly, with at least one ligand selected from the group consisting of:

medicinal substances, antisense probes, gene repair agents, agents for blocking a protein activity and agents for inhibiting a protein activity.

12. A therapeutic composition comprising the conjugate as defined in claim 11.

13. The particle as claimed in claim 7, characterized in that the reactive functional groups are selected from the group consisting of:

carboxyl groups, amine groups, thiol groups, aldehyde groups, hydroxyl groups, tosyl groups and hydrazine groups.

14. The particle as claimed in claim 1, characterized in that the nanoparticles are composed of inorganic materials selected from the group consisting of:

iron, titanium, cobalt, zinc, copper, manganese, chromium and nickel metal oxides;

magnetizable metal oxides;

magnetite;

hematite;

ferrites; and

alloys of cobalt and alloys of nickel.

15. The particle as claimed in claim 14, characterized in that the inorganic materials are magnetizable metal oxides selected from the group consisting of:

iron oxide, magnetite and hematite.

16. The particle as claimed in claim 14, characterized in that the inorganic materials represent from 5 to 95% by mass, with respect to the total mass of said particle.

17. The particle as claimed in claim 16, characterized in that the inorganic materials represent from 25 to 85% by mass, with respect to the total mass of said particle.

18. The particle as claimed in claim 1, characterized in that the inorganic nanoparticles are stabilized by stabilizing agents selected from the group consisting of:

ionic groups, polymer chains, surface-active agents, functional surface-active agents and mixtures of surface-active agents.

19. The particle as claimed in claim 18, characterized in that the surface-active agents are selected from the group consisting of:

sodium dodecyl sulfate and mixtures of sodium dodecyl sulfate and oleic acid.

20. The particle as claimed in claim 1, characterized in that the polymer matrix is a matrix of hydrophobic vinylaromatic polymer types selected from the group consisting of:

homopolymers of water-insoluble vinylaromatic monomers, copolymers of water-insoluble vinylaromatic monomers with one another, copolymers of water-insoluble vinylaromatic monomers with one another and with other water-insoluble polymerizable monomers and copolymers of water-insoluble vinylaromatic monomers with other water-insoluble polymerizable monomers.

21. The particle as claimed in claim 20, characterized in that the polymer is a crosslinked polymer.

22. The particle as claimed in claim 20, characterized in that the water-insoluble vinylaromatic monomers are selected from the group consisting of:

styrene, methylstyrene, ethylstyrene, tert-butylstyrene and vinyltoluene.

23. The particle as claimed in claim 20, characterized in that the other water-insoluble polymerizable monomers are selected from the group consisting of:

alkyl acrylates and methacrylates, wherein the alkyl group comprises from 3 to 10 carbon atoms;

alkyl esters of ethylenic acids, wherein the ethylenic acid possesses 4 or 5 carbon atoms and the alkyl group possesses 1 to 8 carbon atoms;

methacrylic acids;

styrene derivatives; and

diene compounds.

24. The particle as claimed in claim 1, characterized in that it additionally comprises a radioactive tracer.

25. The particle as claimed in claim 1, characterized in that it exhibits a diameter of the order of approximately 50 to 1000 nm.

26. The particle as claimed in claim 25, characterized in that it exhibits a diameter of the order of approximately 100 to 250 nm.

27. A process for the preparation of the composite particle as claimed in claim 1 for which the hydrophobic part of the amphiphilic copolymer is at least partially immobilized in the polymer matrix, characterized in that

(i) a stable and isodisperse starting emulsion composed of two immiscible phases, a hydrophobic phase A composed of droplets comprising inorganic nanoparticles dispersed in an organic phase comprising a surface-active agent and a hydrophilic phase B in which phase A is dispersed, is present;

(ii) hydrophobic monomers and at least one initiator are introduced into phase B; and

(iii) said hydrophobic monomers are polymerized inside phase A,

said process being characterized in that, before stage (iii), at least one amphiphilic copolymer is additionally introduced into the-phase B.

28. The process as claimed in claim 27, characterized in that the amphiphilic copolymer is introduced before or after stage (ii).

29. The process as claimed in claim 27, characterized in that the amphiphilic copolymer is selected from the group consisting of:

block polymers, sequential polymers, branched polymers and comb polymers.

30. The process as claimed in claim 27, characterized in that the hydrophobic monomers are selected from the group consisting of:

vinylaromatic monomers, mixtures of vinylaromatic monomers and mixtures of vinylaromatic monomers with other water-insoluble polymerizable monomers.

31. The process as claimed in claim 30, characterized in that a hydrophobic crosslinking agent is added to the monomer or to the mixture of monomers.

32. The process as claimed in claim 30, characterized in that the vinylaromatic monomers are selected from the group consisting of:

styrene, methylstyrene, ethylstyrene, tert-butylstyrene and vinyltoluene.

33. The process as claimed in claim 30, characterized in that the other water-insoluble polymerizable monomers are selected from the group consisting of:

alkyl acrylates, alkyl methacrylates, alkyl esters of ethylenic acids, methacrylic acids, styrene derivatives, ethylenic acids and diene compounds.

34. The process as claimed in claim 27, characterized in that the initiator is selected from the group consisting of:

organosoluble initiators, initiators that are weakly soluble in the hydrophilic phase and initiators that are soluble in the hydrophilic phase.

35. The process as claimed in claim 34, characterized in that the organosoluble initiator is an azobis type initiator selected from the group consisting:

2,2′

-azobis(2,4-dimethylvaleronitrile), 2,2′

-azobis(4-methoxy-2,4-dimethylvaleronitrile), 2,2′

-azobis(2-cyclopropylpropionitrile), 2,2′

-azobis(2-methylpropionitrile), 2,2′

-azobis(2-methylbutyronitrile), 1,1′

-azobis(cyclohexane-1-carbonitrile), 1-((1-cyano-1-methylethyl)azo) formamide, 2-phenylazo-4-methoxy-2,4-dimethylvaleronitrile, dimethyl 2,2′

-azobis(2-methylpropionate), 4,4′

-azobis(4-cyanovaleric acid) and 2,2′

-azobis(2-(hydroxymethyl)propionitrile).

36. The process as claimed in claim 34, characterized in that the initiator is a weakly soluble or soluble in the hydrophilic phase radical initiator selected from the group consisting of:

peroxides, hydroperoxides and persulfates.

37. The process as claimed in claim 27, characterized in that the initiator is introduced into the hydrophilic phase either simultaneously with the introduction of the hydrophobic monomers or before or after the introduction of said monomers.

38. The process as claimed in claim 27, characterized in that the polymerization (iii) is carried out by photochemistry.

39. The process as claimed in claim 27, characterized in that the organic phase of phase A comprises an aliphatic or cyclic hydrocarbon selected from the group consisting of:

compounds comprising from 5 to 12 carbon atoms, isomers of compounds comprising from 5 to 12 carbon atoms and mixtures of compounds comprising from 5 to 12 carbon atoms, or the organic phase of phase A comprises all or part of an organic compound that can be polymerized by the radical route selected from the group consisting of;

water-insoluble vinylaromatic monomers and copolymers of water-insoluble vinylaromatic monomers with one another.

40. The process as claimed in claim 39, characterized in that the hydrocarbon is selected from the group consisting of:

pentane, hexane, heptane, octane, nonane, decane, undecane and dodecane, and the organic compound that can be polymerized by the radical route is selected from the group consisting of;

styrene, methylstyrene, ethylstyrene, tert-butylstyrene, vinyltoluene, and copolymers of these monomers with one another.

41. The process as claimed in claim 39, characterized in that the water-insoluble vinylaromatic monomers are selected from the group consisting of:

styrene, methylstyrene, ethylstyrene, tert-butylstyrene and vinyltoluene.

42. The process as claimed in claim 27, characterized in that phase B is an aqueous phase.

43. The process as claimed in claim 27, characterized in that it additionally comprises a stage of functionalization of the composite particle by penetration into phase A of reactive functional groups introduced by weakly hydrophilic monomers capable of polymerizing with the hydrophobic monomers.

44. The process as claimed in claim 43, characterized in that the reactive functional groups are acid groups introduced by weakly hydrophilic monomers selected from the group consisting of:

acrylic acid, methacrylic acid, ethylenic acid and sulphonic acid monomers, or by polymerizable monomers selected from the group consisting of;

water-insoluble vinylaromatic monomers, copolymers of vinylaromatic monomers with one another and copolymers of vinylaromatic monomers with other hydrophobic monomers.

45. The process as claimed in claim 44, characterized in that the water-insoluble vinylaromatic monomers are selected from the group consisting of:

styrene, methylstyrene, ethylstyrene, tert-butylstyrene and vinyltoluene.

46. The process as claimed in claim 43, characterized in that weakly hydrophilic monomers are introduced into the phase A either simultaneously with the introduction of the hydrophobic monomers or before or after the introduction of said monomers.

47. The process as claimed in claim 27, characterized in that it additionally comprises, after the polymerization stage (iii), a stage of complete or partial evaporation of phase A.

48. A process for the preparation of the composite particle as claimed in claim 1 and for which the hydrophobic part of the amphiphilic copolymer is at least partially immobilized on the polymer matrix, characterized in that

(i) a stable and isodisperse starting emulsion composed of two immiscible phases, a hydrophobic phase A composed of droplets comprising inorganic nanoparticles dispersed in an organic phase comprising a surface-active agent and a hydrophilic phase B in which phase A is dispersed, is present;

(ii) hydrophobic monomers and at least one initiator are introduced into phase B; and

(iii) a fraction of said hydrophobic monomers is polymerized inside phase A,

said process being characterized in that, before stage (iii) or after stage (iii), at least one amphiphilic copolymer is additionally introduced into phase B and then, according to a stage (iv), the remaining fraction of the hydrophobic monomers is polymerized at the surface of the polymer matrix.

49. The process as claimed in claim 48, characterized in that, when the amphiphilic copolymer is introduced before stage (iii), it is introduced before or after stage (ii).

50. The process as claimed in claim 48, characterized in that the polymerization (iii) and/or (iv) is carried out by heating to a temperature of approximately 60 to 90°

C.