Colloidal suspension of nanoparticles based on an amphiphilic copolymer

• US 20040138095A1
• Filed: 02/20/2004
• Published: 07/15/2004
• Est. Priority Date: 04/02/2001
• Status: Active Grant

• Current Assignee: Flamel Ireland, Ltd.
• Original Assignee: Flamel Technologies SA (Avadel Pharmaceuticals Plc)
• Inventors: Soula, Gerard, Bryson, Nathan
• Granted Patent: US 7,709,445 B2
• Time in Patent Office: Days
• US Class Current: 514/2
• CPC Class Codes: A61K 9/1075 Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers A61K9/0026 takes precedence
• 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: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 5/50 for increasing or potentiating the activity of insulin

Abstract

The invention concerns an aqueous suspension, stable in physiological medium, of nanoparticles for delivering active principles for example insulin. Said delivering particles are based on a three-block copolymer: polyethylene glycol/hydrophilic polyaminoacid/hydrophobic polyaminoacid. Said three-block copolymer particles can be associated with an active principle without denaturing it, and perform a controlled and long-term release of said active principle in vivo, thereby providing the active principle with very prolonged release. The invention also concerns the powder-form solid from which are derived the delivering particles and the preparation of said powder-form solid and said suspension of delivering particles based on three-block copolymer. The invention further concerns pharmaceutical specialities obtainable from said delivering particles filled with active principle.

Claims

1. A colloidal suspension of submicron particles that may be used especially for the vectorization of active principle(s) (AP(s)), these particles being individualized supramolecular arrangements, characterized in that these particles:

are based on at least one amphiphilic copolymer including;

at least one block of hydrophilic polymer(s) of the polyalkylene glycol (PAG) type, preferably polyethylene glycol (PEG); and

at least one linear amphiphilic copolyamino acid (PAA), containing α

-peptide chains; and

are capable of combining in colloidal suspension in undissolved form, with at least one AP and of releasing said AP, especially in vivo, in a sustained and/or delayed manner.

2. The suspension as claimed in claim 1, characterized in that the amino acids of the amphiphilic copolyamino acid(s) (PAA(s)) of which the particles are composed are of at least two types:

a first type comprising at least one hydrophilic amino acid (IAA), preferably selected from the group comprising;

amino acids with one or more ionizable chain(s), which is (are) at least partially ionized, preferably Glu and/or Asp and salts thereof and/or Lys;

mixtures thereof,

a second type comprising at least one hydrophobic amino acid (OAA), preferably selected from the group comprising;

natural neutral amino acids, advantageously those of the sub-group;

Leu, Ile, Val, Ala, Pro, Phe, and mixtures thereof;

rare or synthetic neutral amino acids, advantageously those of the sub-group;

norleucin, norvalin, and mixtures thereof;

derivatives of polar amino acids, advantageously those of the sub-group;

methyl glutamate, ethyl glutamate, benzyl aspartate, N-acetyllysine, and mixtures thereof; and

mixtures thereof.

3. The suspension as claimed in claim 1 or 2, characterized in that the amphiphilic copolyamino acid(s) (PAA(s)) of which the particles are composed include(s) at least one block that is hydrophilic overall and at least one block that is hydrophobic overall.

4. The suspension as claimed in any one of claims 1 to 3, characterized in that the amphiphilic copolyamino acid(s) (PAA(s)) of which the particles are composed has (have) a “

random”

structure.

5. The suspension as claimed in any one of claims 1 to 3, characterized in that the amphiphilic copolyamino acid(s) (PAA(s)) of which the particles are composed has (have) a “

block”

structure.

6. The suspension as claimed in claim 5, characterized in that the “

block”

amphiphilic PAA advantageously includes;

at least one block that is hydrophilic overall, consisting essentially of IAA amino acids, and having an absolute length of greater than or equal to 5 IAA monomers, preferably greater than or equal to 200 IAA monomers and even more preferably between 10 and 50 IAA monomers, and

at least one block that is hydrophobic overall, consisting essentially of OAA amino acids, with an absolute length of greater than or equal to 5 OAA monomers, preferably greater than or equal to 10 OAA monomers and more preferably between 10 and 50 OAA monomers.

7. The suspension as claimed in any one of claims 1 to 6, characterized in that the hydrophilic PAG—

preferably PEG—

is advantageously in the form of a block with an absolute length of greater than or equal to 5 monomers, preferably between 5 and 120 monomers and even more preferably between 5 and 50 monomers.

8. The suspension as claimed in any one of claims 1 to 3 and 5 to 7, characterized in that the particles consist of chains of “

linear triblock”

copolymers PEG/IAA/OAA, preferably corresponding to the following formula;

9. The suspension as claimed in any one of claims 1 to 8, characterized in that the vectorization particles (VPs) have a mean size of between 10 and 500 nm and preferably between 10 and 200 nm.

10. The suspension as claimed in any one of claims 1 to 9, characterized in that the vectorization particles (VPs) include at least one active principle.

11. The suspension as claimed in any one of claims 1 to 10, characterized in that it is aqueous and stable.

12. A pulverulent solid, characterized in that it is obtained from the suspension as claimed in any one of claims 1 to 11.

13. A process for preparing a pulverulent solid obtained from the suspension as claimed in any one of claims 1 to 3 and 5 to 11, characterized in that:

1) at least one PAG block including at least one alkylene glycol monomer is reacted with at least one hydrophilic PAA block comprising at least one hydrophilic amino acid IAA monomer and with at least one hydrophobic PAA block including at least one hydrophobic amino acid OAA monomer, this PAG block and these blocks each including at least one reactive function so as to obtain a PAG/polyIAA/polyOAA “

block”

amphiphilic copolymer;

2) the PAG/polyIAA/polyOAA block amphiphilic copolymer obtained in step 1 is transferred into a medium that is a nonsolvent for the hydrophobic fraction(s) of the amphiphilic copolymer—

preferably into water—

, which leads to the spontaneous formation of PA vectorization particles;

3) optionally, the reaction medium is dialyzed to purify the aqueous suspension of structured particles;

4) optionally, this suspension from step 3 is concentrated;

5) optionally, at least one active principle AP is combined with the particles from step 2, 3 or 4;

6) the liquid medium is removed to collect the pulverulent solid comprising the charged or uncharged particles.

14. A process for preparing the suspension as claimed in any one of claims 1 to 11, characterized in that the pulverulent solid as claimed in claim 12 and/or the pulverulent solid obtained by the process as claimed in claim 13 is placed in contact with an aqueous medium that is a nonsolvent for the hydrophobic fraction of the amphiphilic copolymer.

15. A process for preparing the suspension as claimed in any one of claims 1 to 11, characterized in that it includes steps 1, 2, 3, 4 and optionally 5 of the process as claimed in claim 13.

16. A process for preparing the suspension as claimed in claim 10, characterized in that the AP is combined with the particles by placing a liquid phase containing said hydrophilic AP in contact with the colloidal suspension of particles.

17. A process for preparing the suspension as claimed in claim 11, characterized in that the AP is combined with the particles by placing said AP in solid form in contact with the colloidal suspension of particles.

18. A process for preparing the suspension as claimed in any one of claims 1 to 11, characterized in that the pulverulent solid as claimed in claim 12 and/or the pulverulent solid obtained by the process as claimed in claim 13 is placed in contact with a liquid phase containing the AP.

19. A process for preparing the suspension as claimed in any one of claims 1 to 11, characterized in that the pulverulent solid as claimed in claim 12 and/or the pulverulent solid obtained by the process as claimed in claim 13 is placed in contact with the AP in solid form and in that this mixture of solids is dispersed in a liquid phase, preferably an aqueous solution.

20. A product that is an intermediate of the process as claimed in claim 13, characterized in that it consists of amphiphilic PAA copolymers of the PAG/polyIAA/polyOAA “

block”

type, preferably PEG/polyGlu or Asp/polyONAA, that are particle precursors.

21. The suspension as claimed in any one of claims 1 to 11 and/or obtained by the process as claimed in claim 13 and/or the pulverulent solid as claimed in claim 12, including at least one active principle preferably chosen from:

vaccines, taken alone or combined with at least one antigen;

proteins and/or peptides, among which the ones most preferably selected are;

hemoglobins, cytochromes, albumins, interferons, antigens, antibodies, erythropoietin, insulin, growth hormones, factors VIII and IX, interleukins or mixtures thereof, and hematopoiesis-stimulating factors;

polysaccharides, heparin being more particularly selected;

nucleic acids, and preferably RNA and/or DNA oligonucleotides;

nonpeptide-protein molecules belonging to various anticancer chemotherapy classes, and in particular anthracyclines and taxoids;

and mixtures thereof.

22. A pharmaceutical, nutritional, plant-protection or cosmetic specialty product, characterized in that it includes a suspension as claimed in any one of claims 1 to 11 and/or obtained by the process as claimed in claim 13 and/or the pulverulent solid as claimed in claim 12.