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

Patent metadata

Field	Value
Publication number	US-9475840-B2
Application number	US-201314379484-A
Country	US
Kind code	B2
Filing date	Feb 27, 2013
Priority date	Feb 27, 2012
Publication date	Oct 25, 2016
Grant date	Oct 25, 2016

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)

First claim

The invention claimed is:

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

Assignees

• Univ Barcelona
• Fundació Inst De Recerca Biomèdica (Irb Barcelona)

Inventors

• Giralt Lledó Ernest
• Teixidó Turà Meritxell
• Prades Cosano Roger

Classifications

• A61K39/39533

  against materials from animals · CPC title

• A61K47/48238

  Human Necessities · mapped topic

• A61K49/1866

  the nanoparticle having a (super)(para)magnetic core coated or functionalised with a peptide, e.g. protein, polyamino acid · CPC title

• C07K7/08 Primary

  having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) · CPC title

• A61K49/0056

  Peptides, proteins, polyamino acids · CPC title