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(12)
(54)
(75)
Hung et a].
JP
1050014 A
2/1989
JP
1293314 A
11/1989
JP
3102313 A
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JP
5142502
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6181970
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JP
JP
7223966 A
7324014
W0
W0 94/ 13774
W0
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WO
WO
WO
WO
WO
US 7,125,967 B2
96/20730
97/06782
99/40790
00/30609
02/09513 A2
8/1995
12/1995
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7/1996
2/1997
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6/2000
2/2002
Filed:
Oct. 8, 2003
(65)
Int. Cl.
C08B 37/08
C07H 5/04
(52)
(58)
US 2005/0080245 A1
(51)
OTHER PUBLICATIONS
(2006.01)
(2006.01)
536/20; 536/55.3
536/20,
536/ 55.3
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Bough, W. et al., In?uence of Manufacturing Variables on the
Characteristics and Effectiveness of Chitosan Products. II. Coagu
(Continued)
Primary ExamineriShaojia Anna Jiang
Assistant ExamineriEverett White
(57)
ABSTRACT
2/2002
CN
9/1999
*
8/1987
8/1988
l/l989
38 Claims, No Drawings
US 7,125,967 B2
Page 2
OTHER PUBLICATIONS
Felt, O. et al., Delivery of Antibiotics to the Eye Using a Positively
(1997).
Kristiansen, A. et al., Competitive binding of highly de-N
acetylated chitosans and N,N-diacetylchitobiose to lysoZyrne from
chicken egg White studied by 1H NMRspectroscopy, Carbohydrate
Research, vol. 289, pp. 143-150 (1996).
US 7, 125 ,967 B2
1
20
25
30
Which it is contained.
The resulting chitosans or chitosan derivatives are also
40
55
65
US 7,125 ,967 B2
3
chitins).
Examples of chitosans or chitosan derivatives include, but
are not limited to, chitosan salts; Water-soluble chitosan;
20
25
Endotoxin Concentration
The present invention is directed to methods for making
Water-soluble chitosan or chitosan derivatives (collectively
referred to herein as chitosan or chitosans). The meth
30
40
50
55
60
[A]W[B]X[C]y[D]ZP+Q
Wherein:
each of W, x, y, and Z is independently an integer from 0
to 4 and W+X+y+Z:4;
65
US 7,125 ,967 B2
6
5
A, B, C and D are each independently selected from
wherein:
Illa
lllb
35
lllc
R.\
40
.X/WX
W
folloWing steps:
agitating/mixing the Water-insoluble chitosan in the basic
solution during the basic treatment step;
W2 R3
X
45
X L/XV
50
alkyl, or a halogen;
7.2;
solution;
55
60
dibenZo-l8-croWn-6,
dibenZo-2l-croWn-7,
dibenZo-24
65
US 7,125 ,967 B2
7
20
but are not limited to, acids, such as acetic acid, and
(polyaminopropyl biguanide).
The compositions of the present invention comprise at
30
minutes, more desirably for not less than 1 hour, even more
occurs at room temperature; hoWever, any suitable tempera
60
US 7,125 ,967 B2
9
10
day re-challenge.
ethyleneglyco-bis([3-amino-ethylether)-N,N-tetraacetic
acid.
20
25
EXAMPLE 1
35
include, but are not limited to, surfactants, Which are block
50
Average Amount of
Endotoxin Detected
Sample No.
Sample pH
5.3
45 eu/gram
6.8
45 eu/gram
7.0
45 eu/gram
7.2
45 eu/gram
7.7
45 eu/gram
55
(range of endotoxin)
(from 30460 eu/gram)
(from 30460 eu/gram)
US 7,125 ,967 B2
11
12
EXAMPLE 2
TABLE 2
25
COMPARATIVE EXAMPLE 3
Endotoxin Detected
Sample No.
Sample pH
5.3
(range of endotoxin)
90 eu/gram
6.8
7.0
180 eu/gram
30
7.2
180 eu/gram
7.7
180 eu/gram
35
EXAMPLE 4
Endotoxin Concentration
40
EXAMPLE 5
50
Endotoxin Concentration
Acetylating Step B
EXAMPLE 6
60
65
US 7,125 ,967 B2
14
13
EXAMPLE 7
Endotoxin Concentration
7.0;
adding a Water-miscible solvent into the aqueous solution
having a pH of at least 7.0;
further adjusting the pH of the aqueous solution to a pH
of at least 8.0 to cause precipitation of Water-soluble
TABLE 3
Example No.
3B
3C
4896
12*24
Comparative 3
6000*l 2000
4
5
6
7
12*24
12*24
2448
6*12
tion; and
Washing the Water-soluble chitosan having loW endotoxin
20
EXAMPLE 8
25
30
EXAMPLE 9
35
thereof.
(I)
45
Example No.
8
9
48*96
12*24
Wherein:
each of W, x, y, and Z is independently an integer from 0
to 4 and W+X+y+Z:4;
55
to 4 and W+X+y+Z:4;
US 7,125 ,967 B2
15
16
15. The method of claim 1, Wherein the partially acety
lated Water-soluble chitosan has a degree of N-acetylation of
from about 24% to about 55%, and a degree of O-acetylation
pH adjusting step.
Hla
{\ X
XE X)_>
20
lllb
Water-soluble chitosan.
22. The method of claim 1, Wherein the Water-soluble
chitosan has an endotoxin content of less than about 20
25
XE X)_>
1110
30
.X/WX
@X
R.\
soluble chitosan.
W2 R3
X L/XV
Wherein each X independently represents 0 or S;
45
oXin-free Water.
solvent mixture;
Washing the Water-soluble chitosan having loW endotoxin
content With the Water-miscible solvent; and
wherein:
65
content.
US 7,125 ,967 B2
17
18
(I)
Wherein:
each of W, x, y, and Z is independently an integer from 0
to 4 and W+X+y+Z:4;
soluble chitosan.
38. The method of claim 23, Wherein the Water-soluble
soluble chitosan.