Polypeptide transition metal salts and method of enhancing...

Details Polypeptide transition metal salts and method of enhancing... Polypeptide transition metal salts and method of enhancing...

1999-04-14

2001-12-11

Park, Hankyel T. (Department: 1648)

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

15 to 23 amino acid residues in defined sequence

C424S001170, C424S009200

Reexamination Certificate

active

06329498

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to a transition metal salt of a polypeptide which exhibits a strong affinity to lipopolysaccharides, particularly endotoxins, and moreover the invention relates to a method of enhancing an antiviral activity (for example, an anti-HIV activity) of the polypeptide by which said antiviral activity is expressed stably and strongly by converting said polypeptide to a transition metal salt, and also relates to a pharmaceutical composition or a drug composition for inhibiting a HIV activity, which comprises said transition metal salt of the polypeptide (hereinafter sometimes described as a polypeptide transition metal salt) as an active component.
BACKGROUND OF THE INVENTION
As shown in the publications below, two families of an antimicrobial polypeptide which exhibits an affinity to endotoxins have been isolated from horseshoe crabs.
See, for example, Shigenaga et al., 1990, J. Biol. Chem., 265:21350-21354; Kawano et al., 1990, J. Biol. Chem., 265:15365-15367; Muta et al., 1990, J. Biochem., 108:261-266; Japanese Laid-Open Patent Application No.167230/1990; Japanese Laid-Open Patent Application No.1152987/1990; Japanese Laid-Open Patent Application No.53799/1990; U.S. Pat. No. 5,068,314 (Published Searched Application No.500194/1990); Miyataetal., 1989, J. Biochem., 106:663-668; Akaji et al., 1989, Chem. Pharm. Bull. 37:2661-2664; Tokunaga and Iwanaga, 1989, Taisha(Metabolism), 26:429-439; Shieh et al., 1989, FEBS Lett., 252:121-124; Nakamura et al., 1988, J. Biol. Chem., 263:16709-16713.
One family, a tachyplesin family has been isolated from the Japanese horseshoe crab, Tachypleus. Three tachyplesins, I, II, and III have been identified. Another family, a polyphemusin family has been isolated from the American horseshoe crab,
Limulus polyphemus
. Two polyphemusins, I and II have been identified.
Both families of said tachyplesins and polyphemusins have been found to inhibit growth of both Gram-negative and Gram-positive bacteria at low concentrations as well as fungi, such as
Candida albicans
and form complexes with a bacterial lipopolysaccharide (Shigenaga et al., 1990, J. Biol. Chem., 265:21350-21354; Muta et al., 1990, J. Biochem., 108:261-266).
Also, a polypeptide of the tachyplesin family has been found to exhibit some inhibition activities for virus, such as influenza virus, vesicular stomatitis virus (Murakami et al., 1991, Chemotherapy, 37, 327-334) or human immunodeficiency virus (Morimoto, et al., 1991, Chemotherapy, 37, 206-211).
On the other hand, with respect to the survival of the highly evolved human beings, development of such drugs is extremly longing that are expected to have a prophylactic or therapeutic effect on acquired immune deficiency syndrome (AIDS) caused by infection with human immunodeficiency virus (HIV).
The present inventors have found a series of novel polypeptide which is basically different from the common structure of the polypeptide of horseshoe crabs and exhibits a high antiviral activity against human immunodeficiency virus(HIV) through the studies on the correlation between structural conversion of the polypeptide with endotoxin affinity and the anti-HIV activity, and these results were published in the publications below (Nakashima et al., 1992, Antimicrob. Agents Chemother., 36: 1249-1255; Masuda et al., 1992, Biochem. Biophys. Res. Commun., 189:845-850; Tamamura et al., 1993, Chem. Pharm. Bull., 41:978-980; Tamamura et al., 1993, Biochem. Biophys. Acta, 1163:209-216; Masudaetal., 1992, J. Pharmacobio. Dyn., 15:s-90; U.S. Pat. No. 5,571,892 (International Publication WO 92/04374); U.S. Pat. No. 5,449,752 (Japanese Laid-Open Patent Application No. 163298/1993)).
From results of investigations on structural requirements for expressing an anti-HIV activity of polypeptide based on a basic structure of polypeptide derived from horseshoe crabs, which consists of 16-18 amino acid residues, the present inventors have further provided and filed an improved invention of a novel concept by focusing on minimum essential structure (International Publication WO 95/10534).
According to the aforementioned invention, the structural concept of such lead compound, a polypeptide, which is derived from the polypeptide of the horseshoe crabs as a standard material and exhibits an anti-HIV activity can be summarized into formula [I] below.
1    2    3    4    5    6    7   8   9   10   11   12   13   14   15
[I] (SEQ ID NO: 1)
A
1
- a
2
- Cys - A
2
- A
3
- A
3
- X - Y - Z - A
2
- A
3
- A
3
- Cys - A
3
- A
4
(wherein A
1
independently represents a basic amino acid residue selected from Lys, Arg and Orn; a peptide residue having at least two of said basic amino acid residue; or an N-&agr; substituted amino acid residue or an N-&agr; substituted peptide residue in which a hydrogen atom of N-&agr; position of an amino acid residue in an amino terminus of said basic amino acid residue or said peptide residue may be replaced with an acyl group or a substituted thiocarbamoyl group;
A
2
independently represents an amino acid residue selected from Phe, Trp and Tyr;
A
3
independently represents a basic amino acid residue selected from Arg, Lys and Orn;
A
4
represents an —OH (derived from a carboxyl group) or an —NH
2
(derived from an acid amide group);
X represents a peptide residue of two amino acid residues where at the next position of one amino acid residue selected from Ala, Val, Leu, Ile, Ser, Met and Cys, one of the amino acid of A
2
is connected via a peptide bond;
Y represents a peptide residue of two amino acid residues which consist of a combination of Gly and one amino acid residue selected from A
3
, or a peptide residue of two amino acid residues which consist of a combination of Pro and one amino acid residue selected from D-Arg, D-Lys and D-Orn;
Z represents a peptide residue of two amino acid residues where at the next position of one amino acid residue selected from Ala, Val, Leu, Ile, Ser, Met and A
2
, Cys is connected via a peptide bond;
and X-Y-Z residue connected via peptide bonds is connected to each amino acid residue at the 6th and 10th positions via peptide bonds, or due to the concurrent deletion of X and Z, the residue Y may be connected directly to each amino acid residue at the 6th and 10th positions via peptide bonds, wherein the hydrogen atom of a side chain &ohgr;-amino group of D-Lys, L-Lys, D-Orn or L-Orn which is a constituent amino acid of Y may be substituted with &ohgr;-aminoacyl group).
It has been disclosed already that the polypeptide having a structure of the above-mentioned formula [I] can be provided in addition to the exhibition of its high anti-HIV activity, can maintain the activity by modification of a specific site without lowering the activity, but rather provide a polypeptide of characteristics by the modification which allows a wide variety of selection of physical and/or chemical properties and therapeutic usage that the basic structure has, for example, to increase or decrease its hydrophilicity or lipophilicity, to selectively accumulate it onto a specific tissue, organ or cell, to increase or decrease its retention time in the body, or to develop dosage forms. Among the polypeptide of the formula [I], polypeptides per se which exhibit high anti-HIV activity are exemplified in Table 1.
TABLE 1
Number [I]
1  2  3   4  5  6  7 8 9 10 11 12 13  14 15
A
1
-A
2
-Cys-A
2
-A
3
-A
3
-X-Y-Z-A
2
-A
3
-A
3
-Cys-A
3
-A
4
(SEQ ID NO: 1)

    1    2   3   4   5   6     7       8      &e

Affiliated with

Also associated with

Rating

Polypeptide transition metal salts and method of enhancing... does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Polypeptide transition metal salts and method of enhancing..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Polypeptide transition metal salts and method of enhancing... will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2570650