Protease-resistant compounds useful as shuttles through the blood-brain barrier and shuttle-cargo constructs

{
  "publication_number": "US9475840B2",
  "country": "US",
  "kind": "B2",
  "title": "Protease-resistant compounds useful as shuttles through the blood-brain barrier and shuttle-cargo constructs",
  "abstract": "The peptides of formula (I) where: R 1 is the group attached to the N-terminal of the first amino acid of the sequence P, optionally via the ligand X, and is selected from H, CH 3 C(═O) -, and maleimide; X is a biradical selected from - NH - (CH 2) r - C(═O) -, - C(═O) - (CH 2) r - C(═O) -, - S(CH 2) r -, - S - (CH 2) r - C(═O) -, - O - (CH 2) r -, - S - CH - CH(NH 2) - C(═O) -, - O - (CH 2) r - C(═O) -, - (CH 2) r - C(═O) -, - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, and - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) -; r is 1-5; P is a biradical of an amino acid sequence comprising the sequence D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr (SEQ ID NO: 1); Y is the group attached to the C-terminal of the last amino acid of the sequence P, and is selected from - NH 2, - OH, - OR 2 and - NHR 2; R 2 is a radical selected from (C 1 -C 6)-alkyl and (CH 2) 2 - NH - C(═O) - CH 2 - O - NH 2; k is 0-2; m is 0-1; with the proviso that when the biradical X is - C(═O)(CH 2) r - C(═O) -, then R 1 is H; when the N of the amino acid of the sequence P to which is attached the biradical X is a biradical - NH -, then m is 1, and when is a biradical - N -, then m is 0; and when R 1 is maleimide then the biradical X is - C(═O) - (CH 2) r - C(═O) -, - (CH 2) - C(═O) -, - (CH 2) r - C(═O) -, and - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -. The peptides are useful as shuttles through the blood brain barrier (BBB). The constructs BBB-shuttle-cargo, being the cargo of a biologically active substance or for use in a diagnostic method, are useful in therapy and diagnosis. R 1 - (X) K - P - Y (I)",
  "claims": [
    "1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, R 1 - (X) K - P - Y (I) wherein: R 1 is the group attached to the N-terminal of the first amino acid of the sequence P, optionally via the ligand X, and is selected from the group consisting of H, CH 3 C(═O) -, and maleimide; X is a biradical selected from the group consisting of - NH - (CH 2) r - C(═O) -, - C(═O) - (CH 2) r - C(═O) -, - S(CH 2) r -, - S - (CH 2) r - C(═O) -, - O - (CH 2) r -, - S - CH 2 - CH(NH 2) - C(═O) -, - O - (CH 2) r - C(═O) -, - (CH 2) r - C(═O) -, - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, and - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) -; wherein the biradical X is attached to R 1 and to the N of the sequence P as follows: R 1 - NH - (CH 2) r - C(═O) - N(H) m -, R 1 - C(═O) - (CH 2) r - C(═O) - N(H) m -, R 1 - S - (CH 2) r - N(H) m -, R 1 - S - (CH 2) r - C(═O) - N(H) m -, R 1 - O - (CH 2) r - N(H) m -, R 1 - S - CH 2 - CH(NH 2) - C(═O) - N(H) m, R 1 - O - (CH 2) r - C(═O) - N(H) m -, R 1 - (CH 2) r - C(═O) - N(H) m, R 1 - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, R 1 - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, and R 1 - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) - N(H) m; r is an integer from 1 to 5; P is a biradical of an amino acid sequence comprising the sequence D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr (SEQ ID NO: 1); Y is the group attached to the C-terminal of the last amino acid of the sequence P, and is selected from the group consisting of - NH 2, - OH, - OR 2 and - NHR 2; R 2 is a radical selected from the group consisting of (C 1 -C 6)-alkyl and (CH 2) 2 - NH - C(═O) - CH 2 - O - NH 2; k is an integer from 0 to 2; m is an integer from 0 to 1; with the proviso that when the biradical X is - C(═O)(CH 2) r C(═O) -, then R 1 is H; with the proviso that when m is 1, then the nitrogen of the N-terminal amino acid of the sequence P, which is attached to the biradical X, is in the form of - NH -, and when m is 0, then the nitrogen of the N-terminal amino acid of the sequence P, which is attached to the biradical X, is in the form of “ - N - ”; and with the proviso that when R 1 is maleimide then the biradical X is - C(═O) - (CH 2) r - C(═O) -, - CH(NH 2) - C(═O) -, - (CH 2) r - C(═O) -, and - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -.",
    "2. The peptide compound according to claim 1, wherein: R 1 is the group attached to the N-terminal of the first amino acid of the sequence P, optionally via the ligand X, and is selected from the group consisting of H and CH 3 C(═O) -; X is a biradical selected from the group consisting of - NH - (CH 2) r - C(═O) -, - C(═O) - (CH 2) - C(═O) -, - S(CH 2) r -, - S - (CH 2) r - C(═O) -, - O - (CH 2) r -, - S - CH 2 - CH(NH 2) - C(═O) - and - O - (CH 2) r - C(═O) -; wherein the biradical X is attached to R 1 and to the N of the sequence P as follows: R 1 - NH - (CH 2) r - C(═O) - N(H) m -, R 1 - C(═O) - (CH 2) r - C(═O) - N(H) m -, R 1 - S - (CH 2) r - N(H) m -, R 1 - S - (CH 2) r - C(═O) - N(H) m -, R 1 - O - (CH 2) r - N(H) m -, R 1 - S - CH 2 - CH(NH 2) - C(═O) - N(H) m, and R 1 - O - (CH 2) r - C(═O) - N(H) m -; r is an integer from 1 to 3; P is a biradical of an amino acid sequence comprising the sequence D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr (SEQ ID NO: 1); Y is the group attached to the C-terminal of the last amino acid of the sequence P, and is selected from the group consisting of - NH 2, - OH, - OR 2, and - NHR 2; R 2 is a radical (C 1 -C 6)-alkyl; k is an integer from 0 to 2; m is an integer from 0 to 1; with the proviso that when the biradical X is - C(═O)(CH 2) r C(═O) -, then R 1 is H and with the proviso that when m is 1, then the nitrogen of the N-terminal amino acid of the sequence P, which is attached to the biradical X, is in the form of - NH -, and when m is 0, then the nitrogen of the N-terminal amino acid of the sequence P, which is attached to the biradical X, is in the form of “ - N - ”.",
    "3. The peptide compound according to claim 1, wherein P is a biradical consisting of SEQ ID NO: 1.",
    "4. The peptide compound according to claim 1, where P is a biradical that is the sequence Gly-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr- (SEQ ID NO: 2).",
    "5. The peptide compound according to claim 1, wherein k is an integer from 0 to 1.",
    "6. The peptide compound according to claim 1, wherein Y is selected from - NH 2, - OH and - NHR 2.",
    "7. The peptide compound according to claim 1, which is selected from the group consisting of: H-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (Ia): and H-Gly-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (Ib).",
    "8. A construct of formula (II) or a pharmaceutically acceptable salt thereof, (Z) q - (X) K - P - Y, (II) wherein: X is a biradical selected from the group consisting of - NH - (CH 2) r - C(═O) -, - C(═O) - (CH 2) r - C(═O) -, - S(CH 2) r -, - S - (CH 2) r - C(═O) -, - O - (CH 2) r -, - S - CH 2 - CH(NH 2) - C(═O) -, - O - (CH 2) r - C(═O) -, - (CH 2) r - C(═O) -, - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O), and - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) -; wherein the biradical X is attached to R 1 and to the N of the sequence P as follows: R 1 - NH - (CH 2) r - C═O) - NH m -, R 1 - C(═O) - (CH 2) r - C(═O) - N(H) m -, R 1 - S - (CH 2) r - N(H) m -, R 1 - S - (CH 2) r - C(═O) - N(H) m -, R 1 - O - (CH 2) r - N(H) m -, R 1 - S - CH 2 - CH(NH 2) - C(═O) - N(H) m -, R 1 - O - (CH 2) r - C(═O) - N(H) m -, R 1 - (CH 2) r - C(═O) - N(H) m, R 1 - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, R 1 - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, and R 1 - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) - N(H) m; r is an integer from 1 to 5; P is a biradical of an amino acid sequence comprising the sequence D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-S-Pro-D-Pro-D-Arg-D-His-D-Thr (SEQ ID NO: 1); Y is the group attached to the C-terminal of the last amino acid of the sequence P, and is selected from the group consisting of - NH 2, - OH, - OR 2 and - NHR 2; R 2 is a radical selected from the group consisting of (C 1 -C 6)-alkyl and (CH 2) 2 - NH - C(═O) - CH 2 - 0 - NH 2; k is an integer from 0 to 2; q is an integer from 1 to 2, and Z is a radical of a biologically active substance or a substance for use in a diagnostic method, said substance being substantially unable to cross the BBB by itself; wherein when q is 1, then Z is attached to the N-terminal of the first amino acid of the sequence P, either directly or via the ligand X, and when q is 2, then one Z is attached to the N-terminal of the first amino acid of the sequence P, either directly or via the ligand X, and the other Z is attached either to a nitrogen of the ligand X or to a nitrogen of the lateral chain of the first amino acid of the sequence P.",
    "9. The construct of formula (II) according to claim 8, which has the formula (II′) Z - (X) K - P - Y (II′) wherein: X, P, Y, k are as defined in claim 1, q is 1, X is a biradical selected from the group consisting of - NH - (CH 2) r - C(═O) -, - C(═O) - (CH 2) r - C(═O) -, - S(CH 2) r -, - S - (CH 2) r - C(═O) -, - O - (CH 2) r -, - S - CH 2 - CH(NH 2) - C(═O) -, - O - (CH 2) r - C(═O) -, - (CH 2) r - C(═O) -, - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) -, and - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) -; wherein the biradical X is attached to R 1 and to the N of the sequence P as follows: R 1 - NH - (CH 2) r - C(═O) - N(H) m -, R 1 - C(═O) - (CH 2) r - C(═O) - N(H) m -, R 1 - S - (CH 2) r - N(H) m -, R 1 - S - (CH 2) r - C(═O) - N(H) m -, R 1 - O - (CH 2) r - N(H) m -, R 1 - S - CH 2 - CH(NH 2) - C(═O) - N(H) m, R 1 - O - (CH 2) r - C(═O) - N(H) m -, R 1 - (CH 2) r - C(═O) - N(H) m, R 1 - NH - O - CH 2 - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, R 1 - (CH 2) r - C(═O) - NH - (CH 2) r - CH(NH 2) - C(═O) - N(H) m, and R 1 - NH - (CH 2) r - CH(NHC(═O)CH 2 NH 2) - C(═O) - N(H) m; r is an integer from 1 to 5; P is a biradical of an amino acid sequence comprising the sequence D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr (SEQ ID NO: 1); Y is the group attached to the C-terminal of the last amino acid of the sequence P, and is selected from the group consisting of - NH 2, - OH, - OR 2 and - NHR 2; R 2 is a radical selected from the group consisting of (C 1 -C 6)-alkyl and (CH 2) 2 - NH - CH 2 - O - NH 2; k is an integer from 0 to 2; and Z is a radical of a biologically active substance or a substance for use in a diagnostic method, said substance being substantially unable to cross the BBB by itself.",
    "10. The construct according to claim 8, wherein Z is a radical of an active pharmaceutical ingredient capable of forming an amide bond, an ester bond, a disulfide bond, a thioeter bond, an oxime bond, an amine bond, or an hydrazone bond with X, and is selected from the group consisting of antiretroviral agents, anticancer agents, antipsychotic agents, antineurodegenerative agents, replacement therapy agents, and antiepileptic agents.",
    "11. The construct according to claim 10, wherein the pharmaceutical active agent is selected from the group consisting of an antibody, L-dopamine and H-D-Ala-D-NMePhe-D-Nal(2)-D-Val-D-Leu-D-Lys-D-Lys (SEQ ID NO: 3).",
    "12. The construct according to claim 8, wherein Z is a radical of a compound selected from the group consisting of an antibody, 5(6)-carboxyfluorescein, Quantum dots, and Spion.",
    "13. The construct according to claim 8, which is selected from the group consisting of: L-Dopa-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIa); H-D-Ala-D-NMePhe-D-Nal(2)-D-Val-D-Leu-D-Lys-D-Lys-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIb); 5(6)-carboxifluorescein-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIc); Quantum dot-Gly-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IId); SPION-Gly-D-Pro-D-Trp-D-Val-D-Pro-D-S er-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIe), 5(6)-Carboxifluorescein-L-Lys(Antibody)-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIf); and Rhodamine b-L-Lys(Antibody)-D-Pro-D-Trp-D-Val-D-Pro-D-Ser-D-Trp-D-Met-D-Pro-D-Pro-D-Arg-D-His-D-Thr-NH 2 (IIg).",
    "14. A pharmaceutical composition or a composition for diagnostic purposes comprising a therapeutically effective amount of the construct as defined in claim 8, together with appropriate amounts of pharmaceutically acceptable carriers or excipients and/or acceptable carriers or excipients for diagnosis.",
    "15. A method for the treatment of Parkinson's disease, comprising administering to a mammal in need thereof a therapeutically effective amount of a construct of formula (II) as defined in claim 8, where Z is a radical of dopamine, together with pharmaceutically excipients or carriers.",
    "16. A method for the treatment of Alzheimer's disease, comprising administering to a mammal in need thereof a therapeutically effective amount of a construct of formula (II) as defined in claim 8, wherein Z is a peptidic radical of the peptide H-D-Ala-D-NMePhe-D-Nal(2)-D-Val-D-Leu-D-Lys-D-Lys-NH 2 (SEQ ID NO: 3) together with pharmaceutically excipients or carriers.",
    "17. A method for the diagnosis of a disease comprising administering to a mammal in need of such diagnostic, a therapeutically effective amount of the construct of formula (II) as defined in claim 8 wherein Z is a superparamagentic iron oxide nanoparticle, a gadolinium complex, or a manganese complex, together with one or more pharmaceutically acceptable excipients or carriers and/or acceptable carriers or excipients for diagnosis, generating an image by magnetic resonance imaging, and making a diagnosis using this image.",
    "18. A method for the diagnosis of a disease comprising administering to a mammal in need of such diagnostic, a therapeutically effective amount of the construct of formula (II) as defined in claim 8 wherein Z is selected from a group consisting of 5(6)-carboxyfluorescein, Texas red, and rhodamine and quantum dots, together with one or more pharmaceutically acceptable excipients or carriers and/or acceptable carriers or excipients for diagnosis, generating an image using dual photon microscopy, and making a diagnosis using this image."
  ],
  "description_excerpt": "The present invention relates to the fields of medicine, research and diagnostics, and more specifically to novel compounds that act as shuttles through the blood-brain barrier (BBB) for the delivery of substances that cannot cross the BBB by themselves. It also refers to shuttle-cargo constructs and their use in therapy or diagnostics.\n\nSeveral severe health disorders require treatment of the brain. This includes neurodegenerative diseases such as Parkinson and Alzheimer diseases, but also central nervous system diseases such as schizophrenia, epilepsy or bipolar disorder, brain cancer, human immunodeficiency virus (HIV) and even certain aspects of obesity. The pharmaceutical targets of these diseases are located inside the brain.\n\nThe BBB is a natural filter within the body that regulates the passage substances through from the blood to the brain, allowing only certain substances to cross from the blood to the brain. It is a natural defense mechanism designed to keep harmful substances out of the brain. It controls the composition of the brain extracellular fluid independent of fluctuations within the blood. It is also impermeable for many environmental compounds and drugs.\n\nThe anatomical basis of the BBB is primarily the tight junction at endothelial cells of cerebral microvessels, which form a continuous membrane with no fenestrations. Specific transporters mediate the access of certain molecules important for the brain, such as glucose, isolated amino acids and ions. Other compounds can cross the BBB by a passive diffusion process through the endothelial cells that form the brain microvessels.",
  "cpc": [
    "C07K 7/08",
    "A61K 31/198",
    "A61K 39/39533",
    "A61K 47/48238",
    "A61K 47/48246",
    "A61K 47/62",
    "A61K 47/64",
    "A61K 49/0056",
    "A61K 49/0093",
    "A61K 49/1866"
  ],
  "ipc": [
    "A61K 31/198",
    "A61K 39/395",
    "A61K 47/48",
    "A61K 49/00",
    "A61K 49/18",
    "C07K 7/08"
  ],
  "assignees": [
    "Universitat de Barcelona UB",
    "Fundacio Privada Institut de Recerca Biomedica IRB"
  ],
  "inventors": [
    "Ernest Giralt Lledó",
    "Meritxell Teixidó Turà",
    "Roger Prades Cosano"
  ],
  "filing_date": "2013-02-27",
  "publication_date": "2016-10-25",
  "grant_date": "2016-10-25",
  "priority_date": "2012-02-27",
  "application_number": "US-201314379484-A",
  "family_id": "47884249",
  "cited_by_count": 10,
  "citations": [
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    "WO2007098415A2",
    "US20070231300A1",
    "WO2008025867A1",
    "WO2009008725A2",
    "WO2011009624A1",
    "WO2012007625A1"
  ]
}