Compounds

Details Compounds Compounds

2002-07-19

2004-12-28

Carlson, Karen Cochrane (Department: 1653)

Drug, bio-affecting and body treating compositions

Designated organic active ingredient containing

Peptide containing doai

C530S328000, C424S078270

Reexamination Certificate

active

06835712

ABSTRACT:

CROSS REFERENCES TO RELATED APPLICATIONS
This application is a National Stage Application filed under 35 U.S.C.§371 of PCT/GB00/02364, filed on Jun. 16, 2000.
This invention relates to newly identified spermine:peptide-based surfactant compounds, to the use of such compounds and to processes for their preparation. The invention also relates to the use of the spermine:peptide-based surfactant compounds to facilitate the transfer of compounds into cells for drug delivery.
Surfactants are substances that markedly affect the surface properties of a liquid, even at low concentrations. For example surfactants will significantly reduce surface tension when dissolved in water or aqueous solutions and will reduce interfacial tension between two liquids or a liquid and a solid. This property of surfactant molecules has been widely exploited in industry, particularly in the detergent and oil industries. In the 1970s a new class of surfactant molecule was reported, characterised by two hydrophobic chains with polar heads which are linked by a hydrophobic bridge (Deinega, Y et al.,
Kolloidn. Zh.
36, 649, 1974). These molecules, which have been termed “gemini” (Menger, F M and Littau, C A,
J.Am.Chem.Soc.
113, 1451, 1991), have very desirable properties over their monomeric equivalents. For example they are highly effective in reducing interfacial tension between oil and water based liquids and have a very low critical micelle concentration.
Cationic surfactants have been used inter alia for the transfection of polynucleotides into cells in culture, and there are examples of such agents available commercially to scientists involved in genetic technologies (for example the reagent Tfx™-50 for the transfection of eukaryotic cells available from Promega Corp. WI, USA).
The efficient delivery of DNA to cells in vivo, either for gene therapy or for antisense therapy, has been a major goal for some years. Much attention has concentrated on the use of viruses as delivery vehicles, for example adenoviruses for epithelial cells in the respiratory tract with a view to corrective gene therapy for cystic fibrosis (CF). However, despite some evidence of successful gene transfer in CF patients, the adenovirus route remains problematic due to inflammatory side-effects and limited transient expression of the transferred gene. Several alternative methods for in vivo gene delivery have been investigated, including studies using cationic surfactants. Gao,X et al. (1995)
Gene Ther.
2, 710-722 demonstrated the feasibility of this approach with a normal human gene for CF transmembrane conductance regulator (CFTR) into the respiratory epithelium of CF mice using amine carrying cationic lipids. This group followed up with a liposomal CF gene therapy trial which, although only partially successful, demonstrated the potential for this approach in humans (Caplen, N J. et al.,
Nature Medicine,
1, 39-46, 1995). More recently other groups have investigated the potential of other cationic lipids for gene delivery, for example cholesterol derivatives (Oudrhiri,N et al.
Proc.Natl.Acad.Sci.
94, 1651-1656, 1997). This limited study demonstrated the ability of these cholesterol based compounds to facilitate the transfer of genes into epithelial cells both in vitro and in vivo, thereby lending support to the validity of this general approach.
These studies, and others, show that in this new field of research there is a continuing need to develop novel low-toxicity surfactant molecules to facilitate the effective transfer of polynucleotides into cells both in vitro for transfection in cell-based experimentation and in vivo for gene therapy and antisense treatments. The present invention seeks to overcome the difficulties exhibited by existing compounds.
The invention relates to spermine:peptide-based surfactant compounds having a spermine backbone and having the general structure of formula (I):
where R
1
and R
3
are hydrogen and R
2
and R
4
, which may be the same or different, are peptide groups formed from one or more amino acids linked together, in a linear or branched manner, by amide (CONH) bonds and further linked to the spermine backbone by amide bonds, having the general formula (II):
where p1 is 0 to 5 and p2 is 1 to 5, preferably 1; and the values for p3 and p4, which may be the same or different, are from 0 to 5, preferably 0;
A1, A3 and A4, which may be the same or different, are amino acids selected from serine, lysine, ornithine, threonine, histidine, cysteine, arginine and tyrosine; and
A2 is an amino acid selected from lysine, ornithine and histidine;
and R
5
and R
6
are saturated or unsaturated hydrocarbyl groups having up to 24 carbon atoms and linked to the spermine backbone by an amide or an amine (NCH
2
) linkage;
or
where R
1
and R
3
are hydrogen, R
2
and R
4
, which may be the same or different are saturated or unsaturated hydrocarbyl groups having up to 24 carbon atoms and linked to the spermine backbone by amide or amine bonds, and R
5
and R
6
, which may be the same or different, are peptide groups of formula (II) linked to the spermine backbone by amide bonds;
or
a salt, preferably a pharmaceutically acceptable salt thereof.
When used herein, the term “hydrocarbyl” refers to a group having from 1 to 24 carbon atoms which may be in a straight chain or a branched chain and include a saturated carbocyclic ring having from 3 to 6 carbon atoms and which chain may contain unsaturation (double and/or triple carbon-carbon bonds).
The amide linkages between the amino acids A1, A2 and A3 in the peptide group of formula (II) are standard peptide bonds (&agr; bonds), unless the amino acid is a diamine, for example lysine or ornithine, where the linkage may involve either of the two amine groups. For example, where A1 is lysine, the linkage to the amino acid A2 may be a standard alpha amide bond, or an epsilon (&egr;) amide bond involving the amine of the lysine side chain. Similarly where A1 is ornithine the amide bond linking A1 to A2 may be an alpha bond or a delta (&dgr;) bond that is created using the amine on the side chain of the ornithine amino acid residue.
Preferably, the compound is symmetrical, that is R
1
and R
3
are the same, R
2
and R
4
are the same, and R
5
and R
6
are the same. Symmetrical spermine:peptide-based surfactant compounds of the invention are “gemini” surfactants.
In a preferred embodiment A1 in the group of formula (II) is serine or threonine, prefereably serine. Preferably A3 and A4 in the group of formula (II) are lysine, ornithine, histidine or arginine.
In a further preferred embodiment the hydrocarbyl group is selected from:
—CO(CH
2
)
10
CH
3
—CO(CH
2
)
12
CH
3
—CO(CH
2
)
14
CH
3
—CO(CH
2
)
16
CH
3
—CO(CH
2
)
18
CH
3
—CO(CH
2
)
22
CH
3
—CO(CH
2
)
7
CH═CH(CH
2
)
5
CH
3
—CO(CH
2
)
7
CH═CH(CH
2
)
7
CH
3
—CO(CH
2
)
7
CH═CHCH
2
CH═CH (CH
2
)
4
CH
3
—CO(CH
2
)
7
(CH═CHCH
2
)
3
CH
3
—CO(CH
2
)
3
CH═CH(CH
2
CH═CH)
3
(CH
2
)
4
CH
3
—CO(CH
2
)
7
CH═CH(CH
2
)
5
CH
3
Trans
—CO(CH
2
)
7
CH═CH(CH
2
)
7
CH
3
Trans
—CO(CH
2
)
8
CHCH
3
(CH
2
)
7
CH
3
—COCHOH(CH
2
)
21
CH
3
In another preferred embodiment the hydrocarbyl group is selected from:
—(CH
2
)
11
CH
3
—(CH
2
)
13
CH
3
—(CH
2
)
15
CH
3
—(CH
2
)
17
CH
3
—(CH
2
)
19
CH
3
—(CH
2
)
23
CH
3
—(CH
2
)
8
CH═CH (CH
2
)
5
CH
3
—(CH
2
)
8
CH═CH (CH
2
)
7
CH
3
—(CH
2
)
8
CH═CHCH
2
CH═CH (CH
2
)
4
CH
3
—(CH
2
)
8
(CH═CHCH
2
)
3
CH
3
—(CH
2
)
4
CH═CH(CH
2
CH═CH)
3
(CH
2
)
4
CH
3
—(CH
2
)
8
CH═CH(CH
2
)
5
CH
3
Trans
—(CH
2
)
8
CH═CH(CH
2
)
7
CH
3
Trans
—(CH
2
)
9
CHCH
3
(CH
2
)
7
CH
3
Compounds of the present invention may be prepared from readily available starting materials using synthetic peptide chemistry well known to the skilled person. The scheme shown in
FIGS. 1
a
and
1
b
shows a general process for the synthesis of the compounds of the invention wherein the hydrocarbyl groups are linked to the spermine moiety by amine bonds and the scheme shown in
FIGS. 2
a
and
2
b
shows a general process for

Affiliated with

Also associated with

Rating

Compounds does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Compounds, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Compounds will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3290178