Materials and complexes for the delivery of biologically-active materials to cells

Patent metadata

Field	Value
Publication number	US-9399016-B2
Application number	US-22773507-A
Country	US
Kind code	B2
Filing date	May 30, 2007
Priority date	May 30, 2006
Publication date	Jul 26, 2016
Grant date	Jul 26, 2016

Abstract

The invention provides a peptide derivative of formula A-B-C wherein
A is a polycationic nucleic acid-binding component; B is a spacer element peptide that is susceptible to cleavage within a cell; and C is a cell surface receptor binding component.

The invention also provides a lipid derivative of general formula (I):

(PEG) q -Linker-Spacer-Cationic headgroup-carbon skeleton-(hydrophobic chain) o

wherein:
the Linker is a group susceptible to cleavage within a cell; the Spacer is a group linking the Linker to the Cationic headgroup; q denotes the number of PEG chains and q=1, 2 or 3; o denotes the number of hydrophobic chains and o=1, 2 or 3; the carbon skeleton is a group linking the hydrophobic chains to the cationic headgroup.

The peptide and lipid derivatives find use in non-viral gene delivery systems.

First claim

The invention claimed is:

1. A non-viral delivery complex, for delivering genetic material when combined with a nucleic acid, that comprises:
(i) a peptide of formula A-B-C wherein
A is a polycationic nucleic acid-binding component,
B is a spacer element peptide that is susceptible to cleavage within a cell, and
C is a cell surface receptor binding component,

wherein B, the spacer element peptide comprises amino acid sequences susceptible to cleavage by furin selected from RX 1 KR [SEQ ID NO: 2] and RX 2 RR [SEQ ID NO: 3] in which X 1 and X 2 which may be the same or different and each represent any amino acid residue, or comprises amino acid sequences susceptible to cleavage by cathepsin selected from X 3 X 4 in which X 3 is selected from Tyrosine (Tyr, Y), Phenylalanine (Phe, F), Leucine (Leu, L), Valine (Val, V) and Isoleucine (Ile, I) and X 4 is selected from Glycine (Gly, G), Alanine (Ala, A) and Glutamic acid (Glu, E);
and wherein C, the cell surface receptor binding component, comprises a peptide sequence selected from:
RGD;
RRETAWA [SEQ ID NO: 5];
LDV;
X 5 SM in which X 5 is a basic amino acid residue;
LX 6 HK [SEQ ID NO: 6] in which X 6 is Q or P;
PSGX 7 ARA [SEQ ID NO: 7] in which X 7 is A or T;
SX 8 RSMNF [SEQ ID NO: 8] in which X 8 is an acidic amino acid residue;
LX 9 HKSMP [SEQ ID NO: 9] in which X 9 is P or Q;
PX 10 X 11 X 12 T [SEQ ID NO: 19] in which X 10 , X 11 and X 12 , which may be the same or different, each represents an amino acid residue;
PSX 13 S [SEQ ID NO: 20] in which X 13 represents an amino acid residue;
QX 14 X 15 X 16 Q [SEQ ID NO: 21] in which X 14 and X 16 , which may be the same or different, each represents an amino acid residue, and X 15 represents an amino acid residue having an amide side chain; and
SX 17 S in which X 17 represents an amino acid residue having an aliphatic side chain; and

(ii) a lipid of formula (III):

wherein:
each Y is the same or different and is selected from —O—, and —O—C(O)—;
each R 4 is the same or different and is selected from a straight or branched, saturated or unsaturated C 7-24 hydrocarbyl group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen and OR′, wherein R′ is a C 1-6 hydrocarbyl group;
each R 5 is the same or different and is selected from a straight or branched, saturated or unsaturated C 1-10 hydrocarbyl group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, —OR′, —C(O)OH, —CN, —N(R′) 2 , and —C(O)R′, wherein each R′ is the same or different and is a C 1-6 hydrocarbyl group;
Sp is a C 1-8 alkylene group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen and OR′, wherein R′ is a C 1-6 hydrocarbyl group;
W is selected from —O—C(O)—, —C(O)O—, and the group of formula (IV):

n denotes the number of chains, and n=1 if the linker W is —O—C(O)— or —C(O)O—, and n=2 if the linker W is the group of formula (IV);
each B is the same or different and is a C 1-6 alkylene group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, —OR 1 , —C(O)OH, —CN, —NR 1 R 2 , —C(O)OR 1 , —OC(O)R 1 and —C(O)R 1 ;
R 1 and R 2 are the same or different and are C 1-4 hydrocarbyl;
m is an integer from 1 to 100; and
Q is selected from —N + (R 3 ) 3 , —OH, and —OR 3 wherein R 3 is an unsubstituted C 1-4 alkyl group or a trifluoromethyl group.

2. A non-viral delivery complex, for delivering genetic material when combined with a nucleic acid, that comprises:
(i) a peptide of formula A-B-C wherein
A is a polycationic nucleic acid-binding component,
B is a spacer element peptide that is susceptible to cleavage within a cell, and
C is a cell surface receptor binding component,

wherein B, the spacer element peptide comprises amino acid sequences susceptible to cleavage by furin selected from RX 1 KR [SEQ ID NO: 2] and RX 2 RR [SEQ ID NO: 3] in which X 1 and X 2 which may be the same or different and each represent any amino acid residue, or comprises amino acid sequences susceptible to cleavage by cathepsin selected from X 3 X 4 in which X 3 is selected from Tyrosine (Tyr, Y), Phenylalanine (Phe, F), Leucine (Leu, L), Valine (Val, V) and Isoleucine (Ile, I) and X 4 is selected from Glycine (Gly, G), Alanine (Ala, A) and Glutamic acid (Glu, E);
and wherein C, the cell surface receptor binding component, comprises a peptide sequence selected from:
RGD;
RRETAWA [SEQ ID NO: 5];
LDV;
X 5 SM in which X 5 is a basic amino acid residue;
LX 6 HK [SEQ ID NO: 6] in which X 6 is Q or P;
PSGX 7 ARA [SEQ ID NO: 7] in which X 7 is A or T;
SX 8 RSMNF [SEQ ID NO: 8] in which X 8 is an acidic amino acid residue;
LX 9 HKSMP [SEQ ID NO: 9] in which X 9 is P or Q;
PX 10 X 11 X 12 T [SEQ ID NO: 19] in which X 10 , X 11 and X 12 , which may be the same or different, each represents an amino acid residue;
PSX 13 S [SEQ ID NO: 20] in which X 13 represents an amino acid residue;
QX 14 X 15 X 16 Q [SEQ ID NO: 21] in which X 14 and X 16 , which may be the same or different, each represents an amino acid residue, and X 15 represents an amino acid residue having an amide side chain; and
SX 17 S in which X 17 represents an amino acid residue having an aliphatic side chain; and

(ii) a lipid of formula (VI):

wherein:
each Y is the same or different and is selected from —O—, and —O—C(O)—;
each R 4 is the same or different and is selected from a straight or branched, saturated or unsaturated C 7-24 hydrocarbyl group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen and OR′, wherein R′ is a C 1-6 hydrocarbyl group;
each R 5 is the same or different and is selected from a straight or branched, saturated or unsaturated C 1-10 hydrocarbyl group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, —OR′, —C(O)OH, —CN, —N(R′) 2 , and —C(O)R′, wherein each R′ is the same or different and is a C 1-6 hydrocarbyl group;
Sp is a C 1-8 alkylene group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen and OR′, wherein R′ is a C 1-6 hydrocarbyl group;
W is a group susceptible to cleavage within a cell;
q denotes the number of P-G-chains;
each B is the same or different and is a C 1-6 alkylene group which is unsubstituted or substituted by one or more substituents selected from hydroxy, halogen, —OR 1 , —C(O)OH, —CN, —NR 1 R 2 , —C(O)OR 1 , —OC(O)R 1 and —C(O)R 1 ;
R 1 and R 2 are the same or different and are C 1-4 hydrocarbyl;
p is an integer from 1 to 8; and
Q is selected from —N + (R 3 ) 3 , —OH, and —OR 3 wherein R 3 is an unsubstituted C 1-4 alkyl group or a trifluoromethyl group.

3. A non-viral transfection complex comprising the non-viral delivery complex as claimed in claim 1 or claim 2 and a nucleic acid.

4. A pharmaceutical composition which comprises a transfection complex as claimed in claim 3 in admixture with a pharmaceutically suitable carrier.

5. A method for the treatment of a cancer in a human or in a non-human animal which comprises administering a transfection complex as claimed in claim 3 to the human or to the non-human animal by direct injection into protected tissues or general parenteral administration for non-protected tissues, wherein a t

Assignees

• Hart Stephen Lewis
• Grosse Stephanie
• Tabor Alethea Bernice
• Wong John Bosco
• Elbs Martin
• Hailes Helen Claire
• Mohd Mustapa Mohd Firouz
• Ucl Business Plc

Inventors

• Hart Stephen Lewis
• Grosse Stephanie
• Tabor Alethea Bernice
• Wong John Bosco
• Elbs Martin
• Hailes Helen Claire
• Mohd Mustapa Mohd Firouz

Classifications

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• A61K48/0041

  the non-active part being polymeric · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61K48/0008

  characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition · CPC title

• C07C217/28

  having one amino group and at least two singly-bound oxygen atoms, with at least one being part of an etherified hydroxy group, bound to the carbon skeleton, e.g. ethers of polyhydroxy amines · CPC title