Pharmaceutical compositions comprising viral particles and methods of using the same

Abstract

Provided for herein are pharmaceutical compositions comprising a virus, such as lentiviruses, including pseudotyped lentiviruses and uses thereof. Also provided for herein are methods of using the pharmaceutical compositions.

Description

Abstract This article provides pharmaceutical compositions comprising viruses such as lentiviruses, including pseudolentiviruses, and their uses. Methods of using said pharmaceutical compositions are also provided.

Claims

DOCKET NO: INH-025WO PATENT What is Claimed:

1. A pharmaceutical composition comprising: (i) a lentiviral vector; (ii) a pharmaceutically acceptable buffer at a concentration of about 5 mM to about 100 mM ; (iii) a pharmaceutically acceptable salt at a concentration of about 100 mM to about 800 mM; (iv) a sugar in an amount of about 1% (w / v) to about 5% (w / v); and (v) a protein excipient in an amount of about 0.1% (w / v) to about 3% (w / v), wherein the pharmaceutical composition has a pH of about 6.0 to about 8.

0.

2. The pharmaceutical composition of claim 1, wherein the lentiviral vector is present at a concentration of about 1.0E+05to about 1.0E+09TUs / mL.

3. The pharmaceutical composition of claim 1 or 2, wherein the pharmaceutically acceptable buffer is histidine buffer, HEPES, phosphoric acid buffer, citric acid buffer, acetic acid buffer, succinic acid buffer, phosphate buffer, acetate buffer, citrate buffer, succinate buffer, ascorbic acid buffer, glutamic acid buffer, lactic acid buffer, maleic acid buffer, trometamol buffer, or gluconic acid buffer, or any combination thereof.

4. The pharmaceutical composition of any one of claims 1-3, wherein the pharmaceutically acceptable buffer is a histidine buffer.

5. The pharmaceutical composition of claim 4, wherein the histidine buffer is present at a concentration of about 10 mM to about 30 mM.

6. The pharmaceutical composition of any one of claims 1-5, wherein the pharmaceutical composition has a pH of about 6.5 to about 7.

5.

7. The pharmaceutical composition of any one of claims 1-6, wherein the pharmaceutically acceptable salt is sodium chloride, potassium chloride, magnesium chloride, or calcium chloride, or any combination thereof. -175- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT 8. The pharmaceutical composition of any one of claims 1-7, wherein the pharmaceutically acceptable salt is sodium chloride.

9. The pharmaceutical composition of claim 8, wherein the sodium chloride is present at a concentration of about 100 mM to about 250 mM.

10. The pharmaceutical composition of any one of claims 1-9, wherein the sugar is sucrose or trehalose, or a combination thereof.

11. The pharmaceutical composition of any one of claims 1-10, wherein the sugar is sucrose.

12. The pharmaceutical composition of claim 11, wherein the sucrose is present at about 2% (w / v) to about 4% (w / v).

13. The pharmaceutical composition of any one of claims 1-12, wherein the protein excipient is recombinant human serum albumin, non-recombinant human serum albumin, or a combination thereof.

14. The pharmaceutical composition of any one of claims 1-13, wherein the protein excipient is non-recombinant human serum albumin.

15. The pharmaceutical composition of claim 13 or 14, wherein the non-recombinant human serum albumin is USP grade non-recombinant human serum albumin.

16. The pharmaceutical composition of any one of claims 13-15, wherein the non- recombinant human serum albumin is present at an amount of about 0.1% (w / v), about 0.2% (w / v), about 0.3% (w / v), about 0.4% (w / v), about 0.5% (w / v), about 0.6% (w / v), about 0.7% (w / v), about 0.8% (w / v), about 0.9% (w / v), about 1% (w / v), about 1.1% (w / v), about 1.2% (w / v), about 1.3% (w / v), about 1.4% (w / v), about 1.5% (w / v), about 1.6% (w / v), about 1.7% (w / v), about 1.8% (w / v), about 1.9% (w / v), about 2.0% (w / v), about 2.1% (w / v), about 2.2% (w / v), about 2.3% (w / v), about 2.4% (w / v), about 2.5% (w / v), about 2.6% (w / v), about 2.7% (w / v), about 2.8% (w / v), about 2.9% (w / v), or about 3.0% (w / v).

17. The pharmaceutical composition of any one of claims 1-16, wherein the lentiviral vector -176- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT is a VSV-G pseudotyped lentivirus.

18. The pharmaceutical composition of claim 17, wherein the VSV-G pseudotyped lentivirus comprises a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO:

2.

19. A pharmaceutical composition comprising: (i) a VSV-G pseudotyped lentivirus (ii) about 10 mM to about 30 mM of a L-Histidine buffer; (iii) about 100 mM to about 400 mM of sodium chloride; (iv) about 1% (w / v) to about 5% (w / v) of sucrose; and (v) about 0.1% (w / v) to about 2% (w / v) of a non-recombinant human serum albumin, wherein the pharmaceutical composition has a pH of about 5.5 to about 8.

0.

20. The pharmaceutical composition of claim 19, wherein the VSV-G pseudotyped lentivirus is present at a concentration of about 1.0E+05to 1.0E+09TUs / mL.

21. The pharmaceutical composition of any one of claims 19-20, wherein the L-Histidine is present at a concentration of about 15-25 mM and the pharmaceutical composition has a pH of about 6 to about 7.

22. The pharmaceutical composition of any one of claims 19-21, wherein sodium chloride is present at a concentration of about 125 mM to about 175 mM.

23. The pharmaceutical composition of any one of claims 19-22, wherein sucrose is present in an amount of about 2% (w / v) to about 5% (w / v).

24. The pharmaceutical composition of any one of claims 19-23, wherein the non-recombinant human serum albumin is present at about 0.1% (w / v)to about 1.5% (w / v).

25. The pharmaceutical composition of any one of claims 19-24, wherein the non- recombinant human serum albumin is USP grade non-recombinant human serum albumin.

26. The pharmaceutical composition of any one of claims 19-25, wherein the VSV-G -177- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT pseudotyped lentivirus comprises a VSV-G polypeptide comprising a mutation that corresponds to a mutation at position 182 of SEQ ID NO:

2.

27. The pharmaceutical composition of claims 26, wherein the VSV-G polypeptide comprises a I182A, I182D, or I182E mutation as compared to SEQ ID NO:

2.

28. The pharmaceutical composition of claim 26 or 27, wherein the VSV-G polypeptide comprises a sequence at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to a sequence of SEQ ID NO:

4.

29. The pharmaceutical composition of claim 26 or 27, wherein the VSV-G polypeptide comprises a sequence at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to a sequence of SEQ ID NO:

5.

30. The pharmaceutical composition of any one of claims 26-29, wherein the VSV-G polypeptide, further comprises a mutation that corresponds to a mutation at positions 214 and / or 352 of SEQ ID NO:

2.

31. The pharmaceutical composition of claim 30, wherein the VSV-G polypeptide comprises substitutions at positions I182, T214, and T352 of SEQ ID NO:

2.

32. The pharmaceutical composition of claim 30 or 31, wherein the substitution at position 182 of the VSV-G polypeptide is I182D or I182E, the substitution at position 214 of the VSV- G polypeptide is T214N, and the substitution at position 352 of the VSV-G polypeptide is T352A.

33. The pharmaceutical composition of any one of claims 30-32, wherein the VSV-G polypeptide ,comprises a sequence of SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 24 or SEQ ID NO:

25.

34. A pharmaceutical composition comprising: (i) a VSV-G pseudotyped lentivirus comprising a VSV-G polypeptide comprising a -178- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT mutation that corresponds to a mutation at position 182, 214, and / or 352 of SEQ ID NO: 2; (ii) L-Histidine buffer at a concentration of about 10 mM to about 30 mM; (iii) sodium chloride at a concentration of about 100 mM to about 400 mM; (iv) sucrose in amount of about 1% (w / v) to about 5% (w / v); and (v) non-recombinant human serum albumin in an amount of about 0.1% (w / v) to about 2% (w / v), wherein the pharmaceutical composition has a pH of about 6.5 to about 7.

5.

35. The pharmaceutical composition of any one of claims 1-34, wherein the lentiviral vector further comprises a targeting moiety.

36. The pharmaceutical composition of claim 35, wherein the targeting moiety binds to CD7.

37. The pharmaceutical composition of claim 36, wherein the targeting moiety comprises a polypeptide comprising: (i) a heavy chain variable region comprising heavy chain CDRl, CDR2, and CDR3 sequences, wherein the heavy chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 30; the heavy chain CDR2 has the amino acid sequence of SEQ ID NO: 31; and the heavy chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 32, or variants of any of the foregoing; and (ii) a light chain variable region comprising light chain CDR1, CDR2, and CDR3 sequences, wherein the light chain CDR1 sequence has the amino acid sequence of SEQ ID NO: 33; the light chain CDR2 sequence has the amino acid sequence of SEQ ID NO: 34; and the light chain CDR3 sequence has the amino acid sequence of SEQ ID NO: 35; or variants of any of the foregoing.

38. The pharmaceutical composition of claim 36 or 37, wherein the targeting moiety comprises a heavy chain variable region (VH) comprising an amino acid sequence of SEQ ID NO: 42 and a light chain variable region (VL) comprising an amino acid sequence of SEQ ID NO:

43.

39. The pharmaceutical composition of any one of claims 36-38, wherein the targeting moiety comprises an antigen binding domain comprising an amino acid sequence of SEQ ID NO: 44 or SEQ ID NO:

45. -179- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT 40. The pharmaceutical composition of any one of claims 35-39, wherein the targeting moiety comprises a formula of T-S1, wherein T is a target binding domain and S1 is a stalk portion, wherein the targeting moiety is attached to the surface of the viral vector through a stalk portion, S1,.

41. The pharmaceutical composition of claim 40, wherein the stalk portion, S1, comprises a formula of L1-Fc-L2-X1, wherein: L1 is a linker or absent; Fc is a variant Fc protein; L2 is a linker or absent; and X1 is a polypeptide comprising the transmembrane domain, wherein the targeting moiety having the formula T-S1 has a formula of T-L1-Fc-L2-X1.

42. The pharmaceutical composition of claim 41, wherein X1 comprises a polypeptide having a formula of ECD-TM-ICD, wherein: ECD is an extracellular domain, or a fragment thereof, of a cell surface protein, or absent; TM is a transmembrane domain of a transmembrane protein; and ICD is an intracellular domain or a protein that facilitates incorporation of the targeting moiety into the envelope of the viral particle, or absent, wherein the targeting moiety having the formula of T-L1-Fc-L2-X1 has a formula of T-L1-Fc- L2-ECD-TM-ICD.

43. The pharmaceutical composition of claim 42, wherein: T comprises an amino acid sequence of SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 62, or SEQ ID NO: 63; L1 comprises the amino acid sequence of SEQ ID NO: 72; Fc comprises the amino acid sequence of SEQ ID NO: 82; L2 is absent; ECD comprises the amino acid sequence of SEQ ID NO: 83; TM comprises the amino acid sequence of SEQ ID NO: 84; and ICD comprises an amino acid sequence comprising an env incorporation motif, wherein the env incorporation motif comprises an amino acid sequence of SEQ ID NO:

85. -180- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT 44. The pharmaceutical composition of claim 40, wherein the targeting moiety comprising a formula of T-S1 comprises an amino acid sequence having at least 90% identity to SEQ ID NO:

89.

45. The pharmaceutical composition of any one of claims 1-44, wherein the lentiviral vector further comprises a nucleic acid molecule encoding for a heterologous molecule of interest.

46. The pharmaceutical composition of claim 45, wherein the heterologous molecule of interest is a chimeric antigen receptor (“CAR”).

47. The pharmaceutical composition of claim 46, wherein the CAR comprises an antigen binding domain comprising an antibody, or a fragment thereof.

48. The pharmaceutical composition of claim 47, wherein the antigen binding domain of the CAR binds to CD20.

49. The pharmaceutical composition of claim 48, wherein the antigen binding domain that binds to CD20 comprises a polypeptide comprising a light chain and a heavy chain comprising: a heavy chain variable region of the heavy chain having at least 90% identity to an amino acid sequence of SEQ ID NO: 73; and a light chain variable region of the light chain having at least 90% identity to an amino acid sequence of SEQ ID NO:

74.

50. The pharmaceutical composition of claim 48 or 49, wherein the antigen binding domain that binds to CD20 comprises an amino acid sequence of SEQ ID NO: 75 or SEQ ID NO:

76.

51. The pharmaceutical composition of any one of claims 48-50, wherein the CAR comprises an amino acid sequence having at least 90% identity to SEQ ID NO:

90.

52. A method of infecting a cell, the method comprising contacting the cell with the pharmaceutical composition of any one of claims 1-51.

53. A method of delivering a heterologous molecule of interest to a cell, the method comprising contacting the cell with the pharmaceutical composition of any one of claims 1-51, wherein the lentiviral vector of the pharmaceutical composition comprises a nucleic acid -181- IPTS / 126943357.3DOCKET NO: INH-025WO PATENT molecule encoding the heterologous molecule of interest.

54. A method of delivering a heterologous molecule of interest to a subject, the method comprising administering to the subject the viral particle of any one of claims 1-51, wherein the lentiviral vector of the pharmaceutical composition comprises a nucleic acid molecule encoding the heterologous molecule of interest.

55. A method of treating a disease or disorder in a subject, the method comprising administering to the subject the pharmaceutical composition of any one of claims 1-51, wherein the lentiviral vector of the pharmaceutical composition comprises a nucleic acid molecule encoding a heterologous molecule of interest to treat the disease or disorder.

56. A method of delivering a heterologous molecule to a target cell, the method comprising contacting the cell with the pharmaceutical composition of any one of claims 1-51, wherein the lentiviral vector of the pharmaceutical composition comprises a nucleic acid molecule encoding the heterologous molecule.

57. The pharmaceutical composition of any one of claims 1-51, for use in the treatment of a cancer, an immune disorder, an auto-immune disorder, or a metabolic disorder.

58. Use of the pharmaceutical composition of any one of claims 1-51, for the manufacture of a medicament for treatment of a cancer, an immune disorder, an auto-immune disorder, or a metabolic disorder. -182- IPTS / 126943357.3

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