Compounds for the delivery of granulin across the blood brain barrier

Abstract

Compounds for the delivery of progranulin, a progranulin fragment, and / or at least one granulin protein subunit across the blood brain barrier. Compounds for the delivery of progranulin, a progranulin fragment, and / or at least one granulin protein subunit across the blood brain barrier, the compounds including a progranulin domain, a TfR1 binding domain, and optionally an albumin binding domain. Compounds for the delivery of wildtype or unmodified progranulin across the blood brain barrier.

Description

Abstract of Compounds for Transporting Granulin Across the Blood-Brain Barrier. This abstract describes compounds for transporting pregranulin, pregranulin fragments, and / or at least one granulin subunit across the blood-brain barrier. The compounds comprise a pregranulin domain, a TfR1 binding domain, and optionally an albumin binding domain. The abstract also includes compounds for transporting wild-type or unmodified pregranulin across the blood-brain barrier.

Claims

CLAIMSWe claim:

1. A compound comprising:(a) a progranulin domain; and(b) a transferrin receptor 1 (TfRl) binding domain, wherein the TfRl binding domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions (HCDR) HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions (LCDR) LCDR1, LCDR2, and LCDR3, wherein the amino acid sequence of HCDR1 is SEQ ID NO. 45, the amino acid sequence of HCDR2 is SEQ ID NO. 46, the amino acid sequence of HCDR3 is SEQ ID NO. 47, the amino acid sequence of LCDR1 is SEQ ID NO. 48, the amino acid sequence of LCDR2 is SEQ ID NO. 49, and the amino acid sequence of LCDR3 is SEQ ID NO. 50.

2. The compound of claim 1, wherein the compound further comprises an albumin binding domain, wherein the albumin binding domain the comprises complementarity determining regions (CDR) CDR1, CDR2, and CDR3, wherein the amino acid sequence of CDR1 is SEQ ID NO: 51, the amino acid sequence of CDR2 is SEQ ID NO: 52, and the amino acid sequence of CDR3 is SEQ ID NO: 53.

3. The compound of claim 2, wherein the TfRl binding domain is linked to the progranulin domain with a first linker and the albumin binding domain is linked to the progranulin domain or the TfRl binding domain with a second linker.

4. The compound of claim 2 or 3, wherein the compound is of the formula:Y-Ll-x-L2-zorz-L2-Y-Ll-xandwherein X is the progranulin domain,Y is the TfRl binding domain,Z is the albumin binding domain,Li is the first linker, andL2 is the second linker.

5. The compound of claim 4, wherein the compound is of the formula:Y-LI-X-L2-ZANDwherein the TfRl binding domain is linked to the progranulin domain at an N-terminus of the progranulin domain and the albumin binding domain is linked to the progranulin at a C-terminus of the progranulin domain.

6. The compound of any one of claims 2 to 5, wherein the TfRl binding domain and the albumin binding domain do not form a dimer.

7. The compound of any one of claims 3 to 6, wherein the TfRl binding domain is a Fab region, and the first linker is attached to a heavy chain of the TfRl binding domain or a light chain of the TfRl binding domain.

8. The compound of any one of claims 1 to 7, wherein the TfRl binding domain has an affinity to a human TfRl receptor of from about 1 nM to about 100 nM, from about 5 nM to less than 20 nM, or is about 10 nM.

9. The compound of any one of claims 1 to 8, wherein the TfRl binding domain comprises:(a) a heavy chain, wherein the amino acid sequence of the heavy chain is SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, or SEQ ID NO. 18, and(b) a light chain, wherein the amino acid sequence of the light chain is SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21, or SEQ ID NO. 22.

10. The compound of any one of claims 1 to 9, wherein the TfRl binding domain comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 15 and the amino acid sequence of the LC is SEQ ID NO. 19.

11. The compound of any one of claims 2 to 10, wherein the albumin binding domain is a VHH antibody fragment.

12. The compound of any one of claims 3 to 11, wherein the albumin binding domain is attached to the C-terminus of the progranulin domain through L2.

13. The compound of any one of claims 1 to 22, wherein the amino acid sequence of the albumin binding domain is SEQ ID NO. 23 or SEQ ID NO. 24.

14. The compound of any one of claims 3 to 13, wherein Li and L2 are independently selected from a covalent bond, a peptide linker, a PEG linker, a disulfide bond, a thioacetal linkage, or a thioester linkage.

15. The compound of any one of claims 3 to 14, wherein Li and L2 are identical.

16. The compound of any one of claims 3 to 15, wherein the amino acid sequence of Li and L2are SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO.7, SEQ ID NO. 8, SEQ ID NO.9, SEQ ID NO. 10, SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, or SEQ ID NO.14, preferably wherein the amino acid sequence of Li and L2 are SEQ ID NO. 7.

17. The compound of any one of claims 1 to 16, wherein the progranulin domain is a naturally occurring sequence.

18. The compound of any one of claims 1 to 17, wherein the progranulin domain comprises a fragment of progranulin, wherein the amino sequence of the fragment of progranulin is SEQ ID NO. 2.

19. The compound of any one of claims 1 to 18, wherein the amino acid sequence of the progranulin domain is SEQ ID NO. 1.

20. The compound of any one of claims 1 to 19, wherein the compound comprises a heavy chain with an amino acid sequence having at least 90% sequence identity to the amino acid sequence given by SEQ ID NO. 25, SEQ ID NO. 26, or SEQ ID NO. 29.

21. The compound of any one of claims 1 to 20, wherein the compound comprises a light chain with an amino acid sequence having at least 90% sequence identity to the amino acid sequence given by SEQ ID NO. 19, SEQ ID NO. 32, or SEQ ID NO. 33.

22. The compound of any one of claims 1 to 22, wherein the compound comprises:(a) a heavy chain, wherein the amino acid sequence of the heavy chain is SEQ ID NO. 25, SEQ ID NO. 26, or SEQ ID NO. 29; and(b) a light chain, wherein the amino acid sequence of the light chain is SEQ ID NO. 19, SEQ ID NO. 32, or SEQ ID NO. 33.

23. The compound of any one of claims 1 to 22, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 25 and the amino acid sequence of the LC is SEQ ID NO. 19.

24. The compound of any one of claims 1 to 23, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 26 and the amino acid sequence of the LC is SEQ ID NO. 19.

25. The compound of any one of claims 1 to 24, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 29 and the amino acid sequence of the LC is SEQ ID NO. 32.

26. The compound of any one of claims 1 to 25, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 29 and the amino acid sequence of the LC is SEQ ID NO. 33.

27. A compound comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence given by SEQ ID NO. 25, preferably wherein the compound comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence given by SEQ ID NO. 25.

28. The compound of claim 27, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 25 and the amino acid sequence of the LC is SEQ ID NO. 19.

29. A compound comprising an amino acid sequence having at least 90% sequence identity to the amino acid sequence given by SEQ ID NO. 26, preferably wherein the compound comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence given by SEQ ID NO. 26.

30. The compound of claim 29, wherein the compound comprises a heavy chain (HC) and a light chain (LC), wherein the amino acid sequence of the HC is SEQ ID NO. 26 and the amino acid sequence of the LC is SEQ ID NO. 19.

31. A compound comprising a progranulin fragment, wherein the amino acid sequence of the progranulin fragment is SEQ ID NO. 2.

32. The compound of claim 31, wherein the wherein the progranulin fragment is from 100 residues to 500 residues in length.

33. The compound of any one of claims 1 to 32 for use in the treatment of neuronal ceroid lipofuscinosis type-11, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, or a combination thereof.

34. A method of treating a disorder, the method comprising administering the compound of any one of claims 1 to 32 to a patient in need thereof.

35. The method of claim 34, wherein the disorder is neuronal ceroid lipofuscinosis type-11, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, or a combination thereof.

36. The compound of any one of claims 1 to 32, for use in therapy.

37. A pharmaceutical composition comprising the compound of any one of claims 1 to 32, for use in treating neuronal ceroid lipofuscinosis type-11, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, or a combination thereof.

38. Use of the compound of any one of claims 1 to 32 in the manufacture of a medicament for the treatment of neuronal ceroid lipofuscinosis type-11, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, or a combination thereof.

39. A composition comprising the compound of any one of claims 1 to 32 and a pharmaceutically acceptable carrier.

40. Use of the compound of any one of claims 1 to 32 for the treatment of neuronal ceroid lipofuscinosis type-11, frontotemporal dementia, Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, or a combination thereof.

Images