IMPLANTABLE PARTICLES AND CORRESPONDING METHODS

Claims

1. A particle containing: a) first compartment; b) the second compartment and c) a compound of formula (Ia): (Ia), or its pharmaceutically acceptable salt, where A is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -O-, -C(O)O-, -C(O)-, -OC(O)-, -N(R C )-, -N(R C )C(O)-, -C(O)N(R C )-, -N(R C )N(R D )-, -NCN-, -N(R C )C(O)(C1-C6-alkylene)-, -N(R C )C(O)(C2-C6-alkenylene)-, -C(=N(R C )(R D ))O-, -S-, -S(O) x -, -OS(O) x -, -N(R C )S(O) x -, -S(O) x N(R C )-, -P(R F ) y -, -Si(OR A )2-, -Si(R G )(OR A )-, -B(OR A)- or a metal, each of which is optionally bonded to an addition group (e.g., an addition group described herein) and is optionally substituted with one or more R 1 ; each of L 1 and L 3 independently represents a bond, alkyl, or heteroalkyl, wherein each alkyl and heteroalkyl is optionally substituted with one or more R 2 ; L 2 represents a connection; M is absent or is alkyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted by one or more R 3 ; P is heteroaryl optionally substituted with one or more R 4 ; Z is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl, each of which is optionally substituted with one or more R 5 ; each R A , RB , R C , R D , R E , R F and R G independently represents hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, halogen, azido, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl is optionally substituted with one or more R 6 ; or R C and R D , taken together with the nitrogen atom to which they are attached, form a ring (e.g., a 5-7 membered ring) optionally substituted with one or more R 6 ; each R 1 , R 2 , R 3 , R 4 , R 5 and R 6 independently represents alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, azido, oxo, -OR A1 , -C(O)OR A1 , -C(O)R B1 ,-OC(O)R B1 , -N(R C1 )(R D1 ), -N(R C1 )C(O)R B1 , -C(O)N(R C1 ), SRE1 , S(O) x R E1 , -OS(O) x R E1 , -N(R C1 )S(O) x R E1 , - S(O) x N(R C1 )(R D1 ), -P(R F1 ) y , cycloalkyl, heterocyclyl, aryl, heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is optionally substituted with one or more R 7 ; each R A1 , R B1 , R C1 , R D1 , R E1 and R F1 independently represents hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more R 7 ; each R 7 independently represents alkyl, alkenyl, alkynyl, heteroalkyl, halogen, cyano, oxo, hydroxyl, cycloalkyl, or heterocyclyl; x is 1 or 2; and y is 2, 3, or 4.

2. The particle of claim 1, wherein the first compartment is surrounded by a second compartment.

3. A particle according to any one of claims 1-2, wherein the second compartment forms a barrier around the first compartment.

4. A particle according to any one of claims 1-3, wherein the differential volume of the second compartment is smaller, for example, 1.5 times, 2 times, 3 times or 5 times smaller than the volume of the first compartment.

5. The particle of any one of claims 1-4, wherein the differential volume of the second compartment is approximately 1%, 2%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 75% less than the volume of the first compartment.

6. A particle according to claim 1, characterized by a property selected from the following: a) the first compartment contains a compound of formula (Ia); b) the second compartment contains a compound of formula (Ia); c) the compound of formula (I-a) is located on the outer surface of the particle and / or d) the particle comprises an interface between the first and second compartments, and the compound of formula (Ia) is located at the interface.

7. The particle of claim 1, wherein the first compartment or the second compartment does not substantially contain a compound of formula (Ia).

8. A particle according to claim 1, characterized by a property selected from the following: a) the first compartment does not substantially contain the compound of formula (Ia); b) the second compartment does not substantially contain a compound of formula (Ia); c) the outer surface of the particle is substantially free of the compound of formula (Ia); or d) the particle comprises an interface between the first and second compartments, and the interface does not substantially contain a compound of formula (Ia).

9. A particle according to any one of claims 1 to 8, wherein the particle has a largest linear dimension (LLD), such as a diameter, of from 1 millimeter to 5 millimeters, such as from 1 millimeter to 4 millimeters, from 1 millimeter to 3 millimeters, from 1 millimeter to 2 millimeters, or from 1.5 millimeters to 2 millimeters.

10. A particle according to any one of claims 1-9, wherein the average distance between the outer boundary of the first compartment and the inner boundary of the second compartment is from 500 nanometers to 500 micrometers.

11. A particle according to any one of claims 1-10, wherein the particle comprises a cell.

12. The particle of any one of claims 1-11, wherein the first compartment comprises a cell.

13. A particle according to any one of claims 1-11, wherein the first compartment contains a cell and the second compartment does not contain a cell.

14. A particle according to any one of paragraphs 11-13, wherein the particle contains at least 5×10 6 , 10×10 6 , 15×10 6 or 20×10 6 cells per ml.

15. The particle of any one of claims 11-14, wherein the cell or cells are present as individual cells, one or more spheroids, or associated with one or more microcarriers.

16. A particle according to item 1, where: a) one or a set of cells are located in the first compartment; a) one or a set of cells are located in the second compartment; c) the number or density of cells in the second compartment is at least 2, 5, 10, 10 2 , 10 3 or 10 4 times less than the number or density of cells in the first compartment; d) the first compartment (e.g., the outer boundary of the first compartment) contains a compound of formula (Ia); and / or e) the second compartment (e.g., the outer boundary of the second compartment) contains a compound of formula (Ia).

17. The particle of any one of claims 11-16, wherein the cell is an epithelial cell, an endothelial cell, a fibroblast, a mesenchymal stem cell, or a keratinocyte.

18. The particle of any one of claims 11-17, wherein the cell is an RPE cell (eg, ARPE-19) or an MSC.

19. The particle of any one of claims 11-17, wherein the cell is an islet cell.

20. The particle of any one of claims 11-19, wherein the cell expresses a therapeutic agent (e.g., a polypeptide).

21. The particle of claim 20, wherein the polypeptide is a factor VIII protein or variant thereof (e.g., SEQ ID NO: 1) or a factor IX protein or variant thereof (e.g., SEQ ID NO: 2).

22. The particle of claim 20, wherein the polypeptide is insulin (e.g., insulin A chain, insulin B chain, or proinsulin).

23. A particle according to any one of claims 1-22, wherein the particle comprises a polymer.

24. The particle of claim 23, wherein the polymer is selected from alginate, chitosan, hyaluronate, gelatin, poly(L-lactic acid) (PLLA) or a copolymer of lactic and glycolic acids (PLGA).

25. The particle of any one of claims 23-24, wherein the first compartment comprises a polymer (e.g., a polysaccharide, e.g., alginate).

26. The particle of any one of claims 23-25, wherein the second compartment comprises a polymer (e.g., a polysaccharide, e.g., alginate).

27. A particle according to any one of claims 23-26, wherein the polymers of both the first compartment and the second compartment are modified with a compound of formula (Ia).

28. A particle according to any one of claims 1 to 27, wherein the compound of formula (Ia) is a compound of any of formulas (Ib), (Ic), (Id), (Ie), (If), (II), (II-a), (III), (III-a), (III-b), (III-c) or (III-d) or a pharmaceutically acceptable salt thereof.

29. A particle according to any one of claims 1 to 27, wherein the compound of formula (Ia) is compound 100, compound 101, compound 112, compound 113 or compound 114, as shown in Table 2.

30. The particle of claim 1, wherein the particle is a hydrogel capsule and wherein: a) the first compartment contains a collection of cells engineered to express the polypeptide; b) the first compartment is surrounded by the second compartment; c) the second compartment is essentially cell-free; and d) wherein the second compartment and the outer surface of the particle comprise alginate chemically modified with a compound of formula (Ia).

31. The particle of claim 30, wherein the chemically modified alginate contains a compound of formula (Ia) in an amount that imparts to the particle both anti-fibrotic properties and the necessary mechanical strength.

32. The particle of claim 30 or 31, wherein the compound of formula (Ia) is compound 101, which has the structure:

33. The particle of claim 32, wherein compound 101 is present in the chemically modified alginate at a density of at least 2.0% and less than 9.0% nitrogen (N) (preferably 3.0% to 8.0%, 4.0% to 7.0%, 5.0% to 7.0%, or 6.0% to 7.0%), as determined by combustion analysis to measure the percentage of nitrogen.

34. A particle according to any one of claims 30-31, wherein the first compartment is formed from an alginate solution that does not contain an anti-fibrotic compound (e.g. a compound of formula (Ia)).

35. A particle according to any one of claims 30-34, which has an average diameter from about 1 mm to about 2 mm or an average diameter from about 0.75 to about 1.0 mm.

36. The particle of any one of claims 30-35, wherein the first compartment is formed from an alginate solution containing from about 10 to about 50 million cells / ml, from 50 to about 500 million cells / ml, from about 75 million to about 450 million cells / ml, from about 100 to about 450 million cells / ml, from about 100 to about 400 million cells / ml, or from about 100 to about 300 million cells / ml.

37. The particle of any one of claims 30-36, wherein the cells are derived from ARPE19 cells and comprise an exogenous nucleotide sequence that provides SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:26, SEQ ID NO:27 and SEQ ID NO:

28.

38. The particle of claim 37, wherein the exogenous nucleotide sequence is SEQ ID NO:

23.

39. The particle according to claim 37 or 38, wherein the exogenous nucleotide sequence comprises SEQ ID NO:15 or SEQ ID NO:

27.

40. The particle of claim 37 or 38, wherein the exogenous nucleotide sequence is SEQ ID NO:19 or SEQ ID NO:

28.

41. A particle according to any one of claims 1-40, made by a method comprising contacting a plurality of droplets of a polymer solution with an aqueous crosslinking solution for a period of time sufficient to produce the particle, wherein the crosslinking solution comprises a crosslinking agent, a buffer, and an osmolarity adjusting agent.

42. A preparation based on a plurality of particles, where the plurality comprises a particle according to any one of paragraphs 1-41.

43. The drug according to claim 42, wherein the drug is a pharmaceutically acceptable drug.

44. A method for producing a particle as described herein, for example a particle according to any one of claims 1-40.

45. A particulate composition for use in treating a subject in need of a substance, such as a polypeptide, required by the subject, which comprises: obtaining a particle as described herein, such as in any of embodiments 1-41, that comprises or is capable of producing the substance; and placement of a particle in the subject's body.

Images