Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Nitrogen containing other than solely as a nitrogen in an...
Reexamination Certificate
1997-09-12
2001-08-14
Krass, Frederick (Department: 1614)
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Nitrogen containing other than solely as a nitrogen in an...
C514S256000, C514S668000, C514S669000, C514S670000, C514S674000
Reexamination Certificate
active
06274630
ABSTRACT:
BACKGROUND OF THE INVENTION
The initiation factor, EIf5A, is unique in that it is the only known cellular protein that contains the amino acid hypusine (Hpu) [N
&egr;
-(4-amino-2-hydroxybutyl)lysine], an unusual naturally occurring amino acid, having the structure:
Hypusine was first isolated from bovine brain extracts by Shiba et al in 1971 [Biochim. Biophys. Acta., Vol. 244, pages 523-531 (1971)]. The molecule has two chiral centers at positions 2 and 9, each of which can be classified R or S by the Cahn-Ingold-Prelog method. The post-translational formation of the (2S, 9R) diastereomer:
has been shown to occur on a precursor protein of the eukaryotic initiation factor 5A, i.e., EIf5A (formerly called eIF-4D) [Cooper et al,
Proc. Natl. Acad. Sci. U.S.A
., Vol. 80, pages 1854-1857 (1983); and Safer,
Eur. J. Biochem
., Vol. 186, pages 1-3 (1989)].
EIf5A is biosynthesized by the post-translational aminobutylation of lys-51 of the precursor polypeptide followed by hydroxylation which results in hypusine at residue 51. In the mid-1970's, EIf5A was shown to stimulate ribosomal subunit joining and to enhance 80 S-bound Met-t-RNAi reactivity with puromycin [Anderson et al,
FEBS Lett
., Vol. 76, pages 1-10 (1977); and Kemper et al,
J. Biol. Chem
., Vol. 251, pages 5551-5557 (1976)]. Later in 1983, Cooper et al, supra, suggested that a hypusine-modified protein serves as an important 1-5 initiation factor in all growing eukaryotic cells. In 1986, Park et al [
J. Biol. Chem
., Vol. 261, pages 14515-14519 (1986)] isolated the EIf5A protein from human red blood cells and elucidated the amino acid sequence surrounding the single hypusine residue, as Thr-Gly-Hpu-His-Gly-His-Ala-Lys. EIf5A has also been found to be essential to HIV replication [Bevec et al,
J. Proc. Natl. Acad. Sci. U.S.A
., Vol. 91, pages 10829-10833 (1994); and Ruhl et al,
J. Cell Biol
., Vol. 123, pages 1309-1320 (1994)].
The initial step in the biosynthesis of EIf5A in the cell requires spermidine as the aminobutyl donor.
It is an object of the present invention to provide a method of inhibiting or preventing intracellular biosynthesis of EIf5A.
SUMMARY OF THE INVENTION
The above and other objects are realized by the present invention, one embodiment of which relates to a method for the inhibition or prevention of the intracellular biosynthesis of EIf5A comprising administering to a human or non-human mammal in need thereof an amount of a polyamine sufficient to deplete the supply of intracellular spermidine required for EIf5A biosynthesis, the polyamine having one of the formulae:
wherein: R
1
and R
6
may be the same or different and are H, alkyl or aralkyl having from 1 to 12 carbon atoms, provided that, in formula (I), R
1
and R
6
are not H;
R
2
-R
5
may be the same or different and are H, R
1
or R
6
;
R
7
is H, alkyl, aryl or aralkyl having from 1 to 12 carbon atoms;
m is an integer from 3 to 6, inclusive;
n is an integer from 3 to 6, inclusive;
v, w, x, y and z may be the same or different and are integers from 3 to 10, inclusive;
or its possible stereoisomers wherein:
R
8
-R
13
may be the same or different and are alkyl, branched alkyl, aryl, arylalkyl, cycloalkyl, optionally having an alkyl chain interrupted by at least one etheric oxygen atom, or hydrogen;
N
1
, N
2
, N
3
and N
4
are nitrogen atoms capable of protonation at physiological pH's;
a and b may be the same or different and are integers from 1 to 4, with the proviso that one, but not both, of a and b may be 0;
A, B and C may be the same or different and are bridging groups which effectively maintain the distance between the nitrogen atoms such that the polyamine:
(i) is capable of uptake by a target cell upon administration of the polyamine to a human or non-human mammal or is capable of binding to at least one polyamine site of a receptor located within or on the surface of a cell upon administration of the polyamine to a human or non-human mammal; and
(ii) upon uptake by the target cell, competitively binds via an electrostatic interaction between the positively charged nitrogen atoms to biological counteranions;
the polyamine, upon binding to the biological counter-anion in the cell, functions in a manner biologically different than the intracellular polyamines; and
further wherein at least one of said bridging groups A, B and C may contain at least one —CH(OH)— group which is not alpha- to either of the nitrogen atoms; or
(V) a salt thereof with a pharmaceutically acceptable acid.
Another embodiment of the invention comprises a pharmaceutical composition comprising an amount of a polyamine sufficient, upon administration to a human or non-human mammal in need thereof, to deplete the supply of intracellular spermidine required for EIf5A biosynthesis in the mammal, and a pharmaceutically acceptable carrier therefor, the polyamine having one of the formulae:
wherein: R
1
and R
6
may be the same or different and are H, alkyl or aralkyl having from 1 to 12 carbon atoms, provided that, in formula (I), R
1
and R
6
are not H;
R
2
-R
5
may be the same or different and are H, R
1
or R
6
;
R
7
is H, alkyl, aryl or aralkyl having from 1 to 12 carbon atoms;
m is an integer from 3 to 6, inclusive;
n is an integer from 3 to 6, inclusive;
v, w, x, y and z may be the same or different and are integers from 3 to 10, inclusive;
or its possible stereoisomers wherein:
R
8
-R
13
may be the same or different and are alkyl, branched alkyl, aryl, arylalkyl, cycloalkyl, optionally having an alkyl chain interrupted by at least one etheric oxygen atom, or hydrogen;
N
1
, N
2
, N
3
and N
4
are nitrogen atoms capable of protonation at physiological pH's;
a and b may be the same or different and are integers from 1 to 4, with the proviso that one, but not both, of a and b may be 0;
A, B and C may be the same or different and are bridging groups which effectively maintain the distance between the nitrogen atoms such that the polyamine:
(i) is capable of uptake by a target cell upon administration of the polyamine to a human or non-human mammal or is capable of binding to at least one polyamine site of a receptor located within or on the surface of a cell upon administration of the polyamine to a human or non-human mammal; and
(ii) upon uptake by the target cell, competitively binds via an electrostatic interaction between the positively charged nitrogen atoms to biological counter-anions;
the polyamine, upon binding to the biological counter-anion in the cell, functions in a manner biologically different than the intracellular polyamines; and
further wherein at least one of said bridging groups A, B and C may contain at least one —CH(OH)— group which is not alpha- to either of the nitrogen atoms; or
(V) a salt thereof with a pharmaceutically acceptable acid.
REFERENCES:
patent: 5091576 (1992-02-01), Bergeron
Clarke Dennis P.
Krass Frederick
Miles & Stockbridge
University of Florida Research Foundation Inc.
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