U.S. patent application number 15/563346 was filed with the patent office on 2018-03-22 for biodegradable polyethylene glycol derivative having cyclic benzylidene acetal linker.
This patent application is currently assigned to NOF CORPORATION. The applicant listed for this patent is NOF CORPORATION. Invention is credited to Takuma TSUBUSAKI, Yuji YAMAMOTO.
Application Number | 20180078651 15/563346 |
Document ID | / |
Family ID | 57007176 |
Filed Date | 2018-03-22 |
United States Patent
Application |
20180078651 |
Kind Code |
A1 |
TSUBUSAKI; Takuma ; et
al. |
March 22, 2018 |
BIODEGRADABLE POLYETHYLENE GLYCOL DERIVATIVE HAVING CYCLIC
BENZYLIDENE ACETAL LINKER
Abstract
A biodegradable polyethylene glycol derivative in which a
polyethylene glycol chain is linked by an acetal linker capable of
accurately controlling the hydrolysis rate under different pH
environments in the living body, and whose division rate into a
polyethylene glycol chain of low molecular weight in the living
body can be accurately controlled. The biodegradable polyethylene
glycol derivative is represented by formula (1) or formula (2) as
described.
Inventors: |
TSUBUSAKI; Takuma;
(Kawasaki-shi, Kanagawa, JP) ; YAMAMOTO; Yuji;
(Kawasaki-shi, Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NOF CORPORATION |
Tokyo |
|
JP |
|
|
Assignee: |
NOF CORPORATION
Tokyo
JP
|
Family ID: |
57007176 |
Appl. No.: |
15/563346 |
Filed: |
March 30, 2016 |
PCT Filed: |
March 30, 2016 |
PCT NO: |
PCT/JP2016/060377 |
371 Date: |
September 29, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C08G 65/333 20130101;
C08G 65/33341 20130101; C08G 65/33337 20130101; A61K 47/60
20170801 |
International
Class: |
A61K 47/60 20060101
A61K047/60; C08G 65/333 20060101 C08G065/333 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 31, 2015 |
JP |
2015-070659 |
Claims
1. A biodegradable polyethylene glycol derivative having a cyclic
benzylidene acetal linker represented by formula (1) or formula
(2): ##STR00081## wherein, in the formula (1) and the formula (2),
R.sup.1 and R.sup.6 are each independently a hydrogen atom or a
hydrocarbon group; R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are each
independently an electron-withdrawing or electron-donating
substituent or a hydrogen atom; s is 1 or 2, t is 0 or 1, and s+t
is 1 or 2; P.sup.1 is a straight-chain or branched polyethylene
glycol having a number of ethylene glycol units of 3 or more;
P.sup.2 is a straight-chain or branched polyethylene glycol having
a number of ethylene glycol units of 3 or more; w is an integer of
1 to 8; u is an integer of 1 to 40; v is an integer of 1 to 4;
X.sup.1 is a chemically reactive functional group; and Z.sup.1,
Z.sup.2 and Z.sup.3 are each independently a selected divalent
spacer.
2. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein s is 1 and t is 0, R.sup.2 and R.sup.5 are each a
hydrogen atom, and a sum (.SIGMA..sigma.) of substituent constants
(.sigma.) in R.sup.3, R.sup.4 and P.sup.1--Z.sup.1 in formula (1)
or a sum (.SIGMA..sigma.) of substituent constants (.sigma.) in
R.sup.3, R.sup.4 and P.sup.2--Z.sup.1 in formula (2) satisfies
-0.30.ltoreq..SIGMA..sigma..ltoreq.1.05.
3. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein s is 1 and t is 0, at least one of R.sup.2 and
R.sup.5 is the substituent described above, and a sum
(.SIGMA..sigma.) of substituent constants (.sigma.) in R.sup.3,
R.sup.4 and P.sup.1--Z.sup.1 in formula (1) or a sum
(.SIGMA..sigma.) of substituent constants (.sigma.) in R.sup.3,
R.sup.4 and P.sup.2--Z.sup.1 in formula (2) satisfies
-1.71.ltoreq..SIGMA..sigma..ltoreq.0.88.
4. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein s is 1 and t is 1, or s is 2 and t is 0, R.sup.2
and R.sup.5 are each a hydrogen atom, and a sum (.SIGMA..sigma.) of
substituent constants (.sigma.) in R.sup.3, R.sup.4 and
P.sup.1--Z.sup.1 in formula (1) or a sum (.SIGMA..sigma.) of
substituent constants (.sigma.) in R.sup.3, R.sup.4 and
P.sup.2--Z.sup.1 in formula (2) satisfies
-0.19.ltoreq..SIGMA..sigma..ltoreq.0.57.
5. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein s is 1 and t is 1, or s is 2 and t is 0, at least
one of R.sup.2 and R.sup.5 is the substituent described above, and
a sum (.SIGMA..sigma.) of substituent constants (.sigma.) in
R.sup.3, R.sup.4 and P.sup.1--Z.sup.1 in formula (1) or a sum
(.SIGMA..sigma.) of substituent constants (.sigma.) in R.sup.3,
R.sup.4 and P.sup.2--Z.sup.1 in formula (2) satisfies
-0.98.ltoreq..SIGMA..sigma..ltoreq.0.48.
6. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein w is 1.
7. The biodegradable polyethylene glycol derivative as claimed in
claim 6, wherein P.sup.1 is a straight-chain polyethylene glycol
having a hydrocarbon group or a chemically reactive functional
group at a terminal thereof.
8. The biodegradable polyethylene glycol derivative as claimed in
claim 7, wherein P.sup.1 is represented by formula (3):
Y--(OCH.sub.2CH.sub.2).sub.n-- (3) wherein, in the formula (3), Y
is a hydrocarbon group having from 1 to 24 carbon atoms; and n is
an integer of 3 to 2,000.
9. The biodegradable polyethylene glycol derivative as claimed in
claim 7, wherein P.sup.1 is represented by formula (4):
X.sup.2--Z.sup.4--(OCH.sub.2CH.sub.2).sub.n-- (4) wherein, in the
formula (4), X.sup.2 is a chemically reactive functional group
different from X.sup.1; Z.sup.4 is a divalent spacer; and n is an
integer of 3 to 2,000.
10. The biodegradable polyethylene glycol derivative as claimed in
claim 6, wherein P.sup.1 is a branched polyethylene glycol having a
hydrocarbon group or a chemically reactive functional group
different from X.sup.1 at a terminal thereof.
11. The biodegradable polyethylene glycol derivative as claimed in
claim 10, wherein P.sup.1 is represented by formula (5):
##STR00082## wherein, in the formula (5), Y is a hydrocarbon group
having from 1 to 24 carbon atoms; n is an integer of 3 to 1,000;
and a is 0 or 2.
12. The biodegradable polyethylene glycol derivative as claimed in
claim 10, wherein P.sup.1 is represented by formula (6):
##STR00083## wherein, in the formula (6), X.sup.2 is a chemically
reactive functional group different from X.sup.1; Z.sup.4 is a
divalent spacer; n is an integer of 3 to 1,000, and a is 0 or
2.
13. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein w is 2 to 8.
14. The biodegradable polyethylene glycol derivative as claimed in
claim 13, wherein P.sup.1 is represented by formula (7):
##STR00084## wherein, in the formula (7), X.sup.2 is a chemically
reactive functional group different from X.sup.1; Z.sup.4 is a
divalent spacer; n is an integer of 3 to 1,000, and a is 0 or
2.
15. The biodegradable polyethylene glycol derivative as claimed in
claim 13, wherein P.sup.1 is a straight-chain polyethylene glycol
or a branched polyethylene glycol having a number of terminals of 3
to 8, all terminals of the polyethylene glycol constituting P.sup.1
are each connected to Z.sup.1 in formula (1) or Z.sup.2 in formula
(2), and w is equal to the number of terminals of the polyethylene
glycol.
16. The biodegradable polyethylene glycol derivative as claimed in
claim 15, wherein P.sup.1 is selected from the group consisting of
formula (r), formula (s), formula (t), formula (u) and formula (v):
##STR00085## wherein, in the formulae, n is an integer of 3 to
2,000; and w is 2 in a case where P.sup.1 is represented by formula
(r), w is 3 in a case where P.sup.1 is represented by formula (s),
w is 4 in a case where P.sup.1 is represented by formula (t), w is
4 in a case where P.sup.1 is represented by formula (u), and w is 8
in a case where P.sup.1 is represented by formula (v).
17. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein P.sup.2 is represented by formula (8):
--(OCH.sub.2CH.sub.2).sub.m-- (8) wherein, in the formula (8), m is
an integer of 3 to 2,000; and v in formula (1) or formula (2) is
1.
18. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein P.sup.2 is represented by formula (9):
##STR00086## wherein, in the formula (9), m is an integer of 3 to
1,000, b is 0 or 2; and v in formula (1) or formula (2) is b+2.
19. The biodegradable polyethylene glycol derivative as claimed in
claim 1, wherein X.sup.1 is selected from the group consisting of
an active ester group, an active carbonate group, an aldehyde
group, an isocyanate group, an isothiocyanate group, an epoxy
group, a maleimide group, a vinyl sulfone group, an acryl group, a
sulfonyloxy group, a carboxy group, a thiol group, a dithiopyridyl
group, an .alpha.-haloacetyl group, an alkynyl group, an allyl
group, a vinyl group, an amino group, an oxyamino group, a
hydrazide group and an azide group.
20. The biodegradable polyethylene glycol derivative as claimed in
claim 1 wherein Z.sup.1, Z.sup.2 and Z.sup.3 are each independently
an ether bond, an ester bond, a carbonate bond, a urethane bond, an
amide bond, a secondary amino group, an alkylene group containing
any of these bonds and group, a single bond or an alkylene
group.
21. The biodegradable polyethylene glycol derivative as claimed in
claim 9, wherein X.sup.2 is selected from the group consisting of
an active ester group, an active carbonate group, an aldehyde
group, an isocyanate group, an isothiocyanate group, an epoxy
group, a maleimide group, a vinyl sulfone group, an acryl group, a
sulfonyloxy group, a carboxy group, a thiol group, a dithiopyridyl
group, an .alpha.-haloacetyl group, an alkynyl group, an allyl
group, a vinyl group, an amino group, an oxyamino group, a
hydrazide group and an azide group.
22. The biodegradable polyethylene glycol derivative as claimed in
claim 9, wherein Z.sup.4 is an ether bond, an ester bond, a
carbonate bond, a urethane bond, an amide bond, a secondary amino
group, an alkylene group containing any of these bonds and group, a
single bond or an alkylene group.
Description
TECHNICAL FIELD
[0001] The present invention relates to a biodegradable
polyethylene glycol derivative in which polyethylene glycol chains
are linked by a hydrolysable acetal linker and which is divided in
a living body into a polyethylene glycol chain of low molecular
weight which can be more effectively cleared from the living body.
The polyethylene glycol derivative described in the specification
is used for chemical modification of a biofunctional molecule, for
example, a physiologically active protein, a peptide, an antibody,
a nucleic acid or a low molecular weight drug, or a drug carrier,
for example, a liposome or a polymeric micelle.
BACKGROUND ART
[0002] In drug delivery system, the chemical modification of
biofunctional molecule or drug carrier with polyethylene glycol,
which is a hydrophilic polymer having low antigenicity, is an
effective technique for increasing water solubility and
bioavailability of the drug or the like and for prolonging
circulation time in blood.
[0003] On the other hand, after the drug or the like connected to
such a polyethylene glycol derivative is transported to the tissue
or site as a target to express the efficiency, since the
polyethylene glycol having a large molecular weight is insufficient
in the clearance from the living body, it remains in the body for a
long period of time in some cases.
[0004] As to such a problem, an approach has been made in which the
polyethylene glycol chains are connected with a degradable linker
and the linker is degraded in the living body, thereby dividing the
polyethylene glycol chain into a polyethylene glycol chain of low
molecular weight which can be more effectively cleared from the
living body. Most of the strategies utilize an environment in the
living body, for example, a reductive environment or an act of
degrading enzyme, for the degradation of the linker, and one of
them is a technique of utilizing pH in the living body.
[0005] Under pH environment in the living body, for the purpose of
dividing the polyethylene glycol chain into a polyethylene glycol
chain of low molecular weight which can be more effectively cleared
from the living body, synthesis examples of polyethylene glycol
derivative of division type in which the polyethylene glycol chains
are linked by a hydrolyzable acetal linker have been reported.
[0006] For example, in Patent Document 1, a plurality of
polyethylene glycol derivatives in which two polyethylene glycol
chains are connected through an acetal group derived from various
aldehydes or ketones are disclosed. In Patent Document 1, there is
a disclosure that since the acetal group is hydrolyzed in the
living body so that the polyethylene glycol chain is divided into
two polyethylene glycol chains of low molecular weight, the rate of
clearance from the living body is improved. However, evaluation
data of hydrolysis rate of the acetal group is not shown at all and
also, there is no description on the relevance between the
structure around the acetal group and the hydrolysis rate.
[0007] As described above, although there are examples of
polyethylene glycol derivatives in which the polyethylene glycol
chains are linked by a hydrolysable acetal linker, there is no
example relating to a polyethylene glycol derivative in which the
hydrolysis rate of the acetal linker, that is, the division rate of
the polyethylene glycol chain is accurately controlled.
PRIOR ART DOCUMENT
Patent Document
[0008] Patent Document 1: WO2005/108463
DISCLOSURE OF THE INVENTION
Problems that the Invention is to Solve
[0009] Although pH in the living body varies depending on the site,
the deviation of pH at each site is small. For example, the
periphery of a tumor tissue is an acidic environment in comparison
with pH 7.4 in normal physiological environment, but is weakly
acidic at pH1 of 6.4 to 6.9. Also, the endosome interior and
lysosome interior in the cell have a lower pH, but are at pH of 5.5
to 6.0 and at pH of 4.5 to 5.0, respectively, so that the deviation
of pH is small. Therefore, in order to connect a polyethylene
glycol derivative in which polyethylene glycol chains are linked by
an acetal linker with a drug or the like and after expressing the
efficiency under different pH environments in the living body, to
divide the polyethylene glycol chain into a polyethylene glycol
chain of low molecular weight in each of these sites, it is
necessary to accurately control the hydrolysis rate of the acetal
linker under different pH environments in the living body.
[0010] An object of the present invention is to provide a
biodegradable polyethylene glycol derivative in which polyethylene
glycol chains are linked by an acetal linker capable of accurately
controlling the hydrolysis rate under different pH environments in
the living body, and whose division rate into a polyethylene glycol
chain of low molecular weight in the living body can be accurately
controlled.
Means for Solving the Problems
[0011] As a result of the intensive investigations to solve the
problem described above, the inventors have developed a
biodegradable polyethylene glycol derivative in which polyethylene
glycol chains are linked by a cyclic benzylidene acetal linker
capable of accurately controlling the hydrolysis rate under
different pH environments in the living body, and whose division
rate into a polyethylene glycol chain of low molecular weight in
the living body can be accurately controlled.
[0012] The feature of the invention resides in that a plurality of
polyethylene glycol chains are connected through a cyclic
benzylidene acetal linker having substituent(s). By appropriately
selecting the kind and position of the substituent(s) on the
benzene ring of the cyclic benzylidene acetal linker, the degrees
of electron density and steric hindrance around the acetal group
which affect the hydrolysis rate of the acetal linker can be
adjusted. Based on the feature, it is possible to impart a desired
hydrolysis rate to the acetal linker and after the drug or the like
connected to the biodegradable polyethylene glycol derivative is
transported to the tissue or site as a target to express the
efficiency, it is possible to divide the polyethylene glycol chain
into a polyethylene glycol chain of low molecular weight at an
arbitrary rate under pH environment in each of these sites.
[0013] That is, the invention includes the following items.
[1] A biodegradable polyethylene glycol derivative having a cyclic
benzylidene acetal linker represented by formula (1) or formula (2)
shown below:
##STR00001##
in formula (1) and formula (2), R.sup.1 and R.sup.6 are each
independently a hydrogen atom or a hydrocarbon group; R.sup.2,
R.sup.3, R.sup.4 and R.sup.5 are each independently an
electron-withdrawing or electron-donating substituent or a hydrogen
atom; s is 1 or 2, t is 0 or 1, and s+t is 1 or 2; P.sup.1 is a
straight-chain or branched polyethylene glycol having a number of
ethylene glycol units of 3 or more; P.sup.2 is a straight-chain or
branched polyethylene glycol having a number of ethylene glycol
units of 3 or more; w is a valence of P.sup.1 connected to a cyclic
benzylidene acetal and is an integer of 1 to 8; u is a number of a
structural unit composed of the cyclic benzylidene acetal and
P.sup.2 which are connected each other in series and is an integer
of 1 to 40; v is a number of X.sup.1 connected to P.sup.2 and is an
integer of 1 to 4; X.sup.1 is a chemically reactive functional
group; and Z.sup.1, Z.sup.2 and Z.sup.3 are each independently a
selected divalent spacer. [2] The biodegradable polyethylene glycol
derivative of [1], wherein s is 1 and t is 0, R.sup.2 and R.sup.5
are each a hydrogen atom, and a sum (.SIGMA..sigma.) of substituent
constants (.sigma.) in R.sup.3, R.sup.4 and P.sup.1--Z.sup.1 in
formula (1) or in R.sup.3, R.sup.4 and P.sup.2--Z.sup.1 in formula
(2) satisfies -0.30.ltoreq..SIGMA..sigma..ltoreq.1.05. [3] The
biodegradable polyethylene glycol derivative of [1], wherein s is 1
and t is 0, at least one of R.sup.2 and R.sup.5 is the substituent
described above, and a sum (.sigma.) of substituent constants
(.sigma.) in R.sup.3, R.sup.4 and P.sup.1--Z.sup.1 in formula (1)
or in R.sup.3, R.sup.4 and P.sup.2--Z.sup.1 in formula (2)
satisfies -1.71.ltoreq..SIGMA..sigma..ltoreq.0.88. [4] The
biodegradable polyethylene glycol derivative of [1], wherein s is 1
and t is 1, or s is 2 and t is 0, R.sup.2 and R.sup.5 are each a
hydrogen atom, and a sum (.SIGMA..sigma.) of substituent constants
(.sigma.) in R.sup.3, R.sup.4 and P.sup.1--Z.sup.1 in formula (1)
or in R.sup.3, R.sup.4 and P.sup.2--Z.sup.1 in formula (2)
satisfies -0.19.ltoreq..SIGMA..sigma..ltoreq.0.57. [5] The
biodegradable polyethylene glycol derivative of [1], wherein s is 1
and t is 1, or s is 2 and t is 0, at least one of R.sup.2 and
R.sup.5 is the substituent described above, and a sum
(.SIGMA..sigma.) of substituent constants (.sigma.) in R.sup.3,
R.sup.4 and P.sup.1--Z.sup.1 in formula (1) or in R.sup.3, R.sup.4
and P.sup.2--Z.sup.1 in formula (2) satisfies
-0.98.ltoreq..SIGMA..sigma..ltoreq.0.48. [6] The biodegradable
polyethylene glycol derivative of any one of [1] to [5], wherein w
is 1. [7] The biodegradable polyethylene glycol derivative of [6],
wherein P.sup.1 is a straight-chain polyethylene glycol having a
hydrocarbon group or a chemically reactive functional group at a
terminal thereof. [8] The biodegradable polyethylene glycol
derivative of [7], wherein P.sup.1 is represented by formula
(3):
Y--(OCH.sub.2CH.sub.2).sub.n-- (3)
in the formula (3), Y is a hydrocarbon group having from 1 to 24
carbon atoms; and n is an integer of 3 to 2,000. [9] The
biodegradable polyethylene glycol derivative of [7], wherein
P.sup.1 is represented by formula (4):
X.sup.2--Z.sup.4--(OCH.sub.2CH.sub.2).sub.n-- (4)
in the formula (4), X.sup.2 is a chemically reactive functional
group different from X.sup.1; Z.sup.4 is a divalent spacer; and n
is an integer of 3 to 2,000. [10] The biodegradable polyethylene
glycol derivative of [6], wherein P.sup.1 is a branched
polyethylene glycol having a hydrocarbon group or a chemically
reactive functional group different from X.sup.1 at a terminal
thereof. [11] The biodegradable polyethylene glycol derivative of
[10], wherein P.sup.1 is represented by formula (5):
##STR00002##
in the formula (5), Y is a hydrocarbon group having from 1 to 24
carbon atoms; n is an integer of 3 to 1,000; and a is 0 or 2. [12]
The biodegradable polyethylene glycol derivative of [10], wherein
P.sup.1 is represented by formula (6):
##STR00003##
in the formula (6), X.sup.2 is a chemically reactive functional
group different from X.sup.1; Z.sup.4 is a divalent spacer; n is an
integer of 3 to 1,000, and a is 0 or 2. [13] The biodegradable
polyethylene glycol derivative of any one of [1] to [5], wherein w
is 2 to 8. [14] The biodegradable polyethylene glycol derivative of
[13], wherein P.sup.1 is represented by formula (7):
##STR00004##
in the formula (7), X.sup.2 is a chemically reactive functional
group different from X.sup.1; Z.sup.4 is a divalent spacer, n is an
integer of 3 to 1,000, and a is 0 or 2. [15] The biodegradable
polyethylene glycol derivative of [13], wherein P.sup.1 is a
straight-chain polyethylene glycol or a branched polyethylene
glycol having a number of terminals of 3 to 8, all terminals of the
polyethylene glycol constituting P.sup.1 are each connected to
Z.sup.1 in formula (1) or Z.sup.2 in formula (2), and w is equal to
the number of terminals of the polyethylene glycol. [16] The
biodegradable polyethylene glycol derivative of [15], wherein
P.sup.1 is selected from the group consisting of formula (r),
formula (s), formula (t), formula (u) and formula (v):
##STR00005##
in the formulae, n is an integer of 3 to 2,000; and w is 2 in a
case where P.sup.1 is represented by formula (r), w is 3 in a case
where P.sup.1 is represented by formula (s), w is 4 in a case where
P.sup.1 is represented by formula (t), w is 4 in a case where
P.sup.1 is represented by formula (u), and w is 8 in a case where
P.sup.1 is represented by formula (v). [17] The biodegradable
polyethylene glycol derivative of any one of [1] to [16], wherein
P.sup.2 is represented by formula (8):
--(OCH.sub.2CH.sub.2).sub.m-- (8)
in the formula (8), m is an integer of 3 to 2,000; and v in formula
(1) or formula (2) is 1. [18] The biodegradable polyethylene glycol
derivative of any one of [1] to [16], wherein P.sup.2 is
represented by formula (9):
##STR00006##
in the formula (9), m is an integer of 3 to 1,000, b is 0 or 2; and
v in formula (1) or formula (2) is b+2. [19] The biodegradable
polyethylene glycol derivative of any one of [1] to [18], wherein
X.sup.1 is selected from the group consisting of an active ester
group, an active carbonate group, an aldehyde group, an isocyanate
group, an isothiocyanate group, an epoxy group, a maleimide group,
a vinyl sulfone group, an acryl group, a sulfonyloxy group, a
carboxy group, a thiol group, a dithiopyridyl group, an
.alpha.-haloacetyl group, an alkynyl group, an allyl group, a vinyl
group, an amino group, an oxyamino group, a hydrazide group and an
azide group. [20] The biodegradable polyethylene glycol derivative
of any one of [1] to [19], wherein Z.sup.1, Z.sup.2 and Z.sup.3 are
each independently an ether bond, an ester bond, a carbonate bond,
a urethane bond, an amide bond, a secondary amino group, an
alkylene group containing any of these bonds and group, a single
bond or an alkylene group. [21] The biodegradable polyethylene
glycol derivative of [9], [12] or [14], wherein X.sup.2 is selected
from the group consisting of an active ester group, an active
carbonate group, an aldehyde group, an isocyanate group, an
isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group, a carboxy
group, a thiol group, a dithiopyridyl group, an .alpha.-haloacetyl
group, an alkynyl group, an allyl group, a vinyl group, an amino
group, an oxyamino group, a hydrazide group and an azide group.
[22] The biodegradable polyethylene glycol derivative of [9], [12]
or [14], wherein Z.sup.4 is an ether bond, an ester bond, a
carbonate bond, a urethane bond, an amide bond, a secondary amino
group, an alkylene group containing any of these bonds and group, a
single bond or an alkylene group.
Advantage of the Invention
[0014] In the biodegradable polyethylene glycol derivative having a
cyclic benzylidene acetal linker according to the invention, the
hydrolysis rate of the cyclic benzylidene acetal linker can be
adjusted under different pH environments in the living body.
Therefore, after the drug or the like connected to the
biodegradable polyethylene glycol derivative is transported to the
tissue or site as a target to express the efficiency, it is
possible to divide the polyethylene glycol chain into a
polyethylene glycol chain of low molecular weight at an arbitrary
rate under pH environment in each of these sites. Thus, the
problem, which is a disadvantage in conventional polyethylene
glycol modification, in that since the polyethylene glycol having a
large molecular weight is insufficient in the clearance from the
living body, it remains in the body for a long period of time, can
be fundamentally eliminated. That is, by using the biodegradable
polyethylene glycol derivative in the chemical modification of the
drug or the like, it is able to impart not only the advantages of
polyethylene glycol modification, for example, an increase in water
solubility and bioavailability and prolongation of circulation time
in blood, but also the advantage in that after the drug or the like
expresses the efficiency, the clearance of the polyethylene glycol
from the living body is excellent.
BRIEF DESCRIPTION OF THE DRAWINGS
[0015] FIG. 1 shows results of the hydrolysis test in MES
deuterated water buffer of pD 5.5 at 37.degree. C. using the
compounds of formula (35), formula (44), formula (45), formula (47)
and formula (48) described in Examples.
[0016] FIG. 2 shows results of the hydrolysis test in HEPES
deuterated water buffer of pD 7.4 at 37.degree. C. using the
compounds of formula (35), formula (44), formula (45), formula (47)
and formula (48) described in Examples.
[0017] FIG. 3 shows results of the hydrolysis test in MES
deuterated water buffer of pD 5.5 at 37.degree. C. using the
compounds of formula (41), formula (54), formula (74) and formula
(76) described in Examples.
[0018] FIG. 4 shows results of the hydrolysis test in HEPES
deuterated water buffer of pD 7.4 at 37.degree. C. using the
compounds of formula (41), formula (54), formula (74) and formula
(76) described in Examples.
MODE FOR CARRYING OUT THE INVENTION
[0019] The invention will be described in detail hereinafter.
[0020] The term "acetal" as used in the specification means both of
an acetal structure derived from an aldehyde and an acetal
structure derived from a ketone, that is, a ketal structure.
[0021] The term "cyclic acetal" as used in the invention means both
of a 1,3-dioxolane structure of a 5-membered ring which is s is 1
and t is 0 in formula (1) or formula (2) and a 1,3-dioxane
structure of a 6-membered ring which is s is 1 and t is 1 or s is 2
and t is 0 in formula (1) or formula (2).
[0022] Each of R.sup.1 and R.sup.6 in formula (1) or formula (2) of
the invention is a hydrogen atom or a hydrocarbon group, and a
number of carbon atoms of the hydrocarbon group is preferably 10 or
less and more preferably 4 or less. Specific examples of the
hydrocarbon group include a methyl group, an ethyl group, a propyl
group, an isopropyl group, a tert-butyl group, a phenyl group and a
benzyl group. A preferred embodiment of R.sup.1 is a hydrogen atom
or a methyl group, and a hydrogen atom is more preferred.
[0023] The benzene ring in formula (1) or formula (2) of the
invention may have a plurality of substituents. By appropriately
selecting the kind, the position and the degree of
electron-donating property and electron-withdrawing property of the
substituents on the benzene ring, it is possible to adjust the
degrees of electron density and steric hindrance around the acetal
group which affects the hydrolysis rate of the cyclic acetal
linker. This makes it possible to impart a desired hydrolysis rate
to the cyclic acetal linker.
[0024] In the specification, the substituent(s) on the benzene ring
in formula (1) or formula (2) is described using the "substituent
constant (.sigma.)" which means the substituent constant in the
Hammett's rule which quantifies the effect of the substituent on
the reaction rate or equilibrium of benzene derivative. However, as
is known, the Hammett's rule is applied only to a para-substituted
or meta-substituted benzene derivative and cannot be applied to an
ortho-substituted benzene derivative which is affected by steric
hindrance. Therefore, in the case of ortho-substituted benzene
derivative, the substituent constant means the substituent constant
in the Taft's equation which extends the Hammett's rule described
above.
[0025] In the para-substituted or meta-substituted benzene
derivative described above, the Hammett's rule is represented by
equation (10) shown below.
log(k/k.sub.0)=.rho..sigma. (10)
in the equation, k is a rate constant or equilibrium constant in an
arbitrary reaction of para-substituted or meta-substituted benzene
derivative, k.sub.0 is a rate constant or equilibrium constant in
the case where the benzene derivative does not have the
substituent, that is, the substituent is a hydrogen atom, .rho. is
a reaction constant, and .sigma. is a substituent constant.
[0026] The reaction constant (.rho.) in equation (10) described
above is a constant which is determined depending on reaction
conditions, for example, kind, temperature or solvent of the
reaction, and can be calculated from the slope of Hammett plots. In
the acid hydrolysis reaction of the hydrophilic polymer derivative
having a cyclic benzylidene acetal linker of the invention, in the
case of 1,3-dioxolane structure, the constant is calculated as
".rho.=-2.7" from the results of the hydrolysis tests performed for
the compounds of formula (35), formula (44) and formula (45). Also,
in the case of 1,3-dioxane structure, the constant is calculated as
".rho.=-4.8" from the results of the hydrolysis tests performed for
the compounds of formula (47) and formula (48).
[0027] The substituent constant (.sigma.) in equation (10)
described above is a constant which is determined only depending on
the kind and position of the substituent, regardless of the kind of
reaction, and in the case where no substituent is present, that is,
the substituent is a hydrogen atom, the constant is "0". The term
"electron-withdrawing" as used in the specification means the case
where .sigma. is a positive value and the term "electron-donating"
means the case where .sigma. is a negative value.
[0028] As described above, the Hammett's rule is applied only to a
para-substituted or meta-substituted benzene derivative and cannot
be applied to the case of an ortho-substituted benzene derivative
which is affected by steric hindrance. Therefore, it is the Taft's
equation that the effect of such steric hindrance is introduced as
a factor of the position, that is, a position constant (Es) of the
substituent, to extend the Hammett's rule so that it can also be
applied to the case of the ortho-substituted benzene derivative.
The Taft's equation is represented by equation (11) shown
below.
log(k/k.sub.0)=.rho.*.sigma.*+Es (11)
in the equation, k is a rate constant or equilibrium constant in an
arbitrary reaction of para-substituted or meta-substituted benzene
derivative, k.sub.0 is a rate constant or equilibrium constant in
the case where the benzene derivative does not have a substituent,
that is, the substituent is a hydrogen atom, .rho.* is a reaction
constant, .sigma.* is a substituent constant, and Es is a position
constant of the substituent.
[0029] As is known, since the reaction constant (.rho.) of
para-substituted or meta-substituted benzene derivative and the
reaction constant (.rho.*) of ortho-substituted benzene derivative
are approximately equal, it is defined in the specification that
.rho. and .rho.* are the same. Since the substituent constant
(.sigma.*) in the ortho position is similar to the substituent
constant in the para position as described, for example, in
"Charton, M. Can. J. Chem. 1960, 38, 2493-2499", to the substituent
constant in the ortho position in the specification is applied a
corresponding substituent constant in the para position.
[0030] The substituent constant (.sigma.) in the para position or
the meta position is described in "Hansch, C.; Leo, A.; Taft. R. W.
Chem. Rev. 1991, 91, 165-195", and with respect to a substituent in
which the substituent constant (.sigma.) is unknown, the constant
can be measured and determined by the method described in "Hammett.
L. P. Chem. Rev. 1935, 17(1), 125-136". Moreover, the position
constant (Es) is described in "Unger, S. H.; Hansch, C. Prog. Phys.
Org. Chem. 1976, 12, 91-118". However, as to Es as used in the
specification, a hydrogen atom is defined as "0".
[0031] In formula (1) or formula (2), in the case where a plurality
of substituents are present on the benzene ring, it is defined that
additivity is established for the substituent constant (.sigma.)
and the position constant (Es) thereof, and the sum of .sigma. is
represented by ".SIGMA..sigma." and the sum of Es is represented by
".SIGMA.Es".
[0032] Z.sup.1 is connected to the benzene ring of the cyclic
benzylidene acetal and P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 is also
a substituent of the benzene ring. The substituent constant of
P--Z.sup.1 or P.sup.2--Z.sup.1 can be determined by separately
measuring as to the combination of P.sup.1 and Z.sup.1 or the
combination of P.sup.2 and Z.sup.1, but, since the substituent
constant of P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 is substantially
affected largely by the structure in the vicinity of the connecting
portion to the benzene ring, the effect of the other portions is so
small as to be ignored. Therefore, it is possible to use a known
substituent constant of a structure similar to the structure in the
vicinity of the connecting portion to the benzene ring in place of
separately measuring the substituent constant as to
P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1.
[0033] It is defined that the substituent constant of
P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 in the specification can be
substituted with a substituent constant of a structure in which
atom(s) other than the second atom connected to the third atom
counted from the atom connected to the benzene ring in the backbone
atoms of the main chain of P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 are
substituted with hydrogen atom(s). However, in the case where, when
the atom is substituted with a hydrogen atom, a carboxy group is
formed, it is defined that the substituent constant of
P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 can be substituted with a
substituent constant of a structure in which the atom is
substituted with a methyl group in place of a hydrogen atom.
[0034] Specific examples of the structure of the connecting portion
to the benzene ring in P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 and the
structure for the substitution are shown below. In the case of (r1)
shown below, wherein the connecting portion to the benzene ring in
P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 is an ether bond, a
substituent constant of (r2) shown below is applied. In the cases
of (r3) and (r5) shown below, wherein the connecting portion to the
benzene ring in P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1 is an amide
bond, substituent constants of (r4) and (r6) shown below are
applied, respectively. In the case of (r7) shown below, wherein the
connecting portion to the benzene ring in P.sup.1--Z.sup.1 or
P.sup.2--Z.sup.1 is a urethane bond, a substituent constant of (r8)
shown below is applied.
TABLE-US-00001 Structure of Connecting Portion to Benzene Ring (r1)
##STR00007## (r3) ##STR00008## (r5) ##STR00009## (r7) ##STR00010##
Structure for Substitution (r2) ##STR00011## (r4) ##STR00012## (r6)
##STR00013## (r8) ##STR00014##
[0035] As to the hydrolysis rate of the biodegradable polyethylene
glycol derivative having a cyclic benzylidene acetal linker of the
invention, hydrolysis half-life (t.sub.1/2) in a buffer at pH 5.5
and 37.degree. C. is preferably in the range from 1 hour to 6
months, more preferably in the range from 1 hour to 1 month, and
still more preferably in the range from 1 hour to 24 hours. In the
specification, using a numerical value derived from the compound of
formula (44) described in Examples in which t.sub.1/2 under the
hydrolysis conditions described above is 12 hours, a preferred
range of the sum (.SIGMA..sigma.) of substituent constants in the
case where a 1,3-dioxolane structure is included is defined. When
log(k/k.sub.0) for the compound of formula (44) is calculated using
equation (10) above, equation (12) shown below is obtained.
However, as defined above, P.sup.1--Z.sup.1 in the compound of
formula (44) is substituted with an ethoxy group
(CH.sub.3CH.sub.2O--).
log(k/k.sub.0)=-2.7.times.(0.34-0.24)=-0.27 (12)
[0036] In the case where R.sup.2 and R in formula (1) or formula
(2) are hydrogen atoms, when log(k'/k.sub.0) is calculated by
taking the rate constant at the time when t.sub.1/2 is 24 hours as
k' using equation (12) and equation (10) above, equation (13) shown
below is obtained.
log(k'/k)=log {(12/24)k/k}=-0.30
[0037] When the equation is modified,
log(k'/k)=log [(k'/k.sub.0)/(k/k.sub.0)]=-0.30
log(k'/k.sub.0)-log(k/k.sub.0)=-0.30
[0038] When equation (12) above is substituted,
log(k'/k.sub.0)-(-0.27)=-0.30
log(k'/k.sub.0)=-0.57 (13)
[0039] Here, when the sum (.SIGMA..sigma.) of the substituent
constants is calculated using equation (13) and equation (10)
above, equation (14) shown below is obtained.
log(k'/k.sub.0)=-2.7.times..SIGMA..sigma.=-0.57
.SIGMA..sigma.=0.21 (14)
[0040] Similarly, in the case where R.sup.2 and R.sup.5 in formula
(1) or formula (2) are hydrogen atoms, when log(k''/k.sub.0) is
calculated by taking the rate constant at the time when t.sub.1/2
is 1 hour as k'', equation (15) shown below is obtained.
log(k''/k)=log(12k/k}=1.08
[0041] When the equation is modified,
log(k''/k)=log [(k''/k.sub.0)/(k/k.sub.0)]=1.08
log(k''/k.sub.0)-log(k/k.sub.0)=1.08
[0042] When equation (12) above is substituted,
log(k''/k.sub.0)-(-0.27)=1.08
log(k''/k.sub.0)=0.81 (15)
[0043] Here, when the sum (.SIGMA..sigma.) of the substituent
constants is calculated using equation (15) and equation (10)
above, equation (16) shown below is obtained.
log(k''/k.sub.0)=-2.7.times..SIGMA..sigma.=0.81
.SIGMA..sigma.=-0.30 (16)
[0044] From equation (14) and equation (16), in the case where
formula (1) or formula (2) includes a 1,3-dioxolane structure and
R.sup.2 and R.sup.5 are hydrogen atoms, when the range of
.SIGMA..sigma. satisfies -0.30.ltoreq..SIGMA..sigma..ltoreq.0.21,
t.sub.1/2 of the biodegradable polyethylene glycol derivative is
represented by 1 hour.ltoreq.t.sub.1/2.ltoreq.24 hours. Similarly,
when the ranges of .SIGMA..sigma. at 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month and 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months are calculated,
-0.30.ltoreq..SIGMA..sigma..ltoreq.0.76 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month and
-0.30.ltoreq..SIGMA..sigma..ltoreq.1.05 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months, respectively.
[0045] The substituent which can be used in the invention is a
substituent which does not inhibit the acetalization reaction of
the cyclic benzylidene acetal linker compound, the coupling
reaction of the cyclic benzylidene acetal linker compound with a
polyethylene glycol intermediate, the terminal functional group
conversion reaction of the polyethylene glycol intermediate and the
linking reaction of the polyethylene glycol intermediate in the
synthesis process of the biodegradable polyethylene glycol
derivative, and further the bond-forming reaction between the
biodegradable polyethylene glycol derivative and the drug or the
like.
[0046] The substituent may be any of electron-withdrawing
substituent and electron-donating substituent as far as it
satisfies the conditions described above, and the substituents may
be used individually or in combination. The electron-withdrawing
substituent includes an acyl group having from 2 to 5 carbon atoms,
an alkoxycarbonyl group having from 2 to 5 carbon atoms, a
carbamoyl group having from 2 to S carbon atoms, an acyloxy group
having from 2 to 5 carbon atoms, an acylamino group having from 2
to 5 carbon atoms, an alkoxycarbonylamino group having from 2 to 5
carbon atoms, a fluorine atom, a chlorine atom, a bromine atom, an
iodine atom, an alkylsulfanyl group having from 1 to 4 carbon
atoms, an alkylsulfonyl group having from 1 to 4 carbon atoms, an
arylsulfonyl group having from 6 to 10 carbon atoms, a nitro group,
a trifluoromethyl group and a cyano group, and preferred examples
thereof include an acetyl group, a methoxycarbonyl group, a
methylcarbamoyl group, an acetoxy group, an acetamide group, a
methoxycarbonylamino group, a fluorine atom, a chlorine atom, a
bromine atom, an iodine atom, a methylsulfanyl group, a
phenylsulfonyl group, a nitro group, a trifluoromethyl group and a
cyano group. The electron-donating substituent includes an alkyl
group having from 1 to 4 carbon atoms, and preferred examples
thereof include a methyl group, an ethyl group, a propyl group, an
isopropyl group and a tert-butyl group. The substituent which is an
electron-withdrawing group in the meta-position and an
electron-donating group in the para-position or the ortho-position
includes an alkoxy group having from 1 to 4 carbon atoms, an aryl
group having from 6 to 10 carbon atom and an aryloxy group having
from 6 to 10 carbon atoms, and preferred examples thereof include a
methoxy group, an ethoxy group, a propoxy group, an isopropoxy
group, a tert-butoxy group, a phenyl group and a phenoxy group.
[0047] In the case where formula (1) or formula (2) includes a
1,3-dioxolane structure and at least one of R.sup.2 and R.sup.5 is
a substituent other than a hydrogen atom, using the position
constants (Es) of a phenyl group which has the largest influence of
steric hindrance and a fluorine atom which has the smallest
influence of steric hindrance among the substituents described
above, the ranges of .SIGMA..sigma. in a buffer at pH 5.5 and
37.degree. C. at 1 hour.ltoreq.t.sub.1/2.ltoreq.24 hours, 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month, and 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months are calculated by using
Taft's equation (11), respectively. As a result, it is found that
-1.71.ltoreq..SIGMA..sigma..ltoreq.0.04 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.24 hours,
-1.71.ltoreq..SIGMA..sigma..ltoreq.0.59 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month, and
-1.71.ltoreq..SIGMA..sigma..ltoreq.0.88 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months, respectively.
[0048] In the case where formula (1) or formula (2) includes a
1,3-dioxolane structure and R.sup.2 and R.sup.5 are hydrogen atoms,
for example, a preferred embodiment which satisfies
-0.30.ltoreq..SIGMA..sigma..ltoreq.0.21 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.24 hours is described below. However,
the substituents shown herein means R.sup.3 and R.sup.4 and the
structure used in place of P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1
according to the definition described above. In the preferred
embodiment, one of the meta-positions in formula (1) or formula (2)
is a methoxy group, an ethoxy group or an acetamide group, and more
preferably an ethoxy group or an acetamide group. In another
preferred embodiment, the para-position in formula (1) or formula
(2) is a methoxy group or an ethoxy group and one of the
meta-positions is a substituent selected from the group consisting
of a fluorine atom, a chlorine atom, a bromine atom and an iodine
atom, and more preferably the para-position is an ethoxy group and
one of the meta-positions is a fluorine atom or a chlorine atom. In
still another preferred embodiment, one of the para-position and
the meta-position in formula (1) or formula (2) is a methoxy group,
an ethoxy group or an acetamide group, and more preferably a
methoxy group or an ethoxy group.
[0049] Further, in the case where formula (1) or formula (2)
includes a 1,3-dioxolane structure and at least one of R.sup.2 and
R.sup.5 is a substituent other than a hydrogen atom, for example, a
preferred embodiment which satisfies
-1.71.ltoreq..SIGMA..sigma..ltoreq.0.04 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.24 hours is described below. However,
the substituents shown herein means R.sup.3 and R.sup.4 and the
structure used in place of P.sup.1--Z.sup.1 or P.sup.2--Z.sup.1
according to the definition described above. In the case where one
of R.sup.2 and R.sup.5 in formula (1) or formula (2) is a fluorine
atom, a methyl group or an ethyl group and the other is a hydrogen
atom, the para-position is preferably an ethoxy group or an
acetamide group, and more preferably an ethoxy group. In the case
where one of R.sup.2 and R.sup.5 in formula (1) or formula (2) is a
methoxy group and the other is a hydrogen atom, the para-position
is preferably a substituent selected from the group consisting of a
methoxymethyl group and an acetamide group, and more preferably an
acetamide group.
[0050] Moreover, using a numerical value derived from the compound
of formula (35) described in Examples in which the hydrolysis
half-life (t.sub.1/2) in a buffer at pH 5.5 and 37.degree. C. is 24
hours, a preferred range of the sum (.SIGMA..sigma.) of substituent
constants in the case where formula (1) or formula (2) includes a
1,3-dioxane structure can be defined.
[0051] In the case where formula (1) or formula (2) includes a
1,3-dioxane structure and R.sup.2 and R.sup.5 are hydrogen atoms,
when the range of .SIGMA..sigma. satisfies
-0.19.ltoreq..SIGMA..sigma..ltoreq.0.10, t.sub.1/2 of the
hydrophilic polymer derivative is represented by 1
hour.ltoreq.t.sub.1/2.ltoreq.24 hours. Similarly, when the ranges
of .SIGMA..sigma. at 1 hour.ltoreq.t.sub.1/2.ltoreq.1 month and 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months are calculated,
-0.19.ltoreq..SIGMA..sigma..ltoreq.0.41 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month and
-0.19.ltoreq..SIGMA..sigma..ltoreq.0.57 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months, respectively.
[0052] Further, in the case where formula (1) or formula (2)
includes a 1,3-dioxone structure and at least one of R.sup.2 and
R.sup.5 is a substituent other than a hydrogen atom, using the
position constants (Es) of a phenyl group which has the largest
influence of steric hindrance and a fluorine atom which has the
smallest influence of steric hindrance among the substituents
described above, the ranges of .SIGMA..sigma. in a buffer at pH 5.5
and 37.degree. C. at 1 hour.ltoreq.t.sub.1/2.ltoreq.24 hours, 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month, and 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months are calculated by using
Taft's equation (11), respectively. As a result, it is found that
-0.98.ltoreq..SIGMA..sigma..ltoreq.0.00 at the time of 1
hour.ltoreq.t.sub.1/2.gtoreq.24 hours,
-0.98.ltoreq..SIGMA..sigma..ltoreq.0.31 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.1 month, and
-0.98.ltoreq..SIGMA..sigma..ltoreq.0.48 at the time of 1
hour.ltoreq.t.sub.1/2.ltoreq.6 months, respectively.
[0053] As described above, the kind and position of the
substituent(s) suitable for imparting the desired hydrolyzability
to the cyclic benzylidene acetal linker in the biodegradable
polyethylene glycol derivative of the invention can be reasonably
set by performing the calculation described above using equation
(10) and equation (11).
[0054] X.sup.1 in formula (1) or formula (2) of the invention is
not particularly limited as long as it is a functional group which
forms a covalent bond upon a reaction with a functional group
present in a biofunctional molecule, for example, a physiologically
active protein, a peptide, an antibody, a nucleic acid or a low
molecular drug, or a drug carrier, for example, a liposome or a
polymeric micelle, which is the object of chemical modification.
For example, the functional groups include those described in
"Harris, J. M. Poly(Ethylene Glycol) Chemistry; Plenum Press: New
York, 1992", "Hermanson, G. T. Bioconjugate Techniques, 2nd ed.;
Academic Press: San Diego, Calif., 2008", "PEGylated Protein Drugs:
Basic Science and Clinical Applications; Veronese, F. M., Ed.;
Birkhauser; Basel, Switzerland, 2009" and the like.
[0055] Preferred examples of X.sup.1 include an active ester group,
an active carbonate group, an aldehyde group, an isocyanate group,
an isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group, a carboxy
group, a thiol group, a dithiopyridyl group, an .alpha.-haloacetyl
group, an alkynyl group, an allyl group, a vinyl group, an amino
group, an oxyamino group, a hydrazide group and an azide group.
[0056] More specifically, the functional group capable of forming a
covalent bond upon a reaction with an amino group of the
biofunctional molecule is an active ester group, an active
carbonate group, an aldehyde group, an isocyanate group, an
isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group or a carboxy
group, the functional group capable of forming a covalent bond upon
a reaction with a thiol group of the biofunctional molecule is an
active ester group, an active carbonate group, an aldehyde group,
an isocyanate group, an isothiocyanate group, an epoxy group, a
maleimide group, a vinyl sulfone group, an acryl group, a
sulfonyloxy group, a carboxy group, a thiol group, a dithiopyridyl
group, an .alpha.-haloacetyl group, an alkynyl group, an allyl
group or a vinyl group, the functional group capable of forming a
covalent bond upon a reaction with an aldehyde group or a carboxy
group of the biofunctional molecule is a thiol group, an amino
group, an oxyamino group or a hydrazide group, the functional group
capable of forming a covalent bond upon a reaction with an alkynyl
group of the biofunctional molecules is a thiol group or an azide
group, and the functional group capable of forming a covalent bond
upon a reaction with an azide group of the biofunctional molecule
is an alkynyl group.
[0057] The term "active ester" as referred to herein indicates an
activated carboxy group represented by formula: --C(.dbd.O)-L,
wherein L represents a leaving group.
[0058] The leaving group represented by L includes a
succinimidyloxy group, a phthalimidyloxy group, a 4-nitrophenoxy
group, a 1-imidazolyl group, a pentafluorophenoxy group, a
benzotriazol-1-yloxy group, a 7-azabenzotriazol-1-yloxy group and
the like. The term "active carbonate" as referred to herein
indicates an activated carbonate group represented by formula:
--O--C(.dbd.O)-L, wherein L represents a leaving group same as
described above.
[0059] In a preferred embodiment of the invention, X.sup.1 is a
group represented by group (I), group (II), group (III), group (IV)
or group (V).
Group (I): Functional group capable of forming a covalent bond upon
a reaction with an amino group of the biofunctional molecule
[0060] (a), (b), (c), (d), (e) and (f) shown below:
Group (II): Functional group capable of forming a covalent bond
upon a reaction with a thiol group of the biofunctional
molecule
[0061] (a), (b), (c), (d), (e), (f), (g), (h), (i) and (j) shown
below:
Group (III): Functional group capable of forming a covalent bond
upon a reaction with an aldehyde group or a carboxy group of the
biofunctional molecule
[0062] (g), (k), (l) and (m) shown below:
Group (IV): Functional group capable of forming a covalent bond
upon a reaction with an alkynyl group of the biofunctional
molecule
[0063] (g), (k), (l), (m) and (n) shown below:
Group (V): Functional group capable of forming a covalent bond upon
a reaction with an azide group of the biofunctional molecule
[0064] (j) shown below:
##STR00015##
[0065] In the formulae above, R.sup.7 is a hydrogen atom or a sulfo
group, specific examples of the sulfo group include sodium
sulfonate and potassium sulfonate, and R.sup.7 is preferably a
hydrogen atom. R.sup.8 and R.sup.11 are each a hydrogen atom or a
hydrocarbon group having from 1 to 5 carbon atoms, and specific
examples of the hydrocarbon group include a methyl group, an ethyl
group, a propyl group, an isopropyl group, a butyl group, a
tert-butyl group and a pentyl group. R.sup.9 is a hydrocarbon group
having from 1 to 10 carbon atoms which may contain a halogen atom,
specific examples of the hydrocarbon group include a methyl group,
an ethyl group, a propyl group, an isopropyl group, a butyl group,
a tert-butyl group, a pentyl group, an isopentyl group, a hexyl
group, a benzyl group, a 4-methylphenyl group, a trifluoromethyl
group, a 2,2,2-trifluoroethyl group, a 4-(trifluoromethoxy)phenyl
group, a vinyl group, a chloroethyl group, a bromoethyl group and
an iodoethyl group, and R.sup.9 is preferably a methyl group, a
vinyl group, a 4-methylphenyl group or a 2,2,2-trifluoroethyl
group. R.sup.10 is a halogen atom selected from the group
consisting of a chlorine atom, a bromine atom and an iodine
atom.
[0066] Z.sup.1 in formula (1) or formula (2) of the invention is a
divalent spacer between the benzene ring of the cyclic benzylidene
acetal group and the polyethylene glycol chain, Z.sup.2 is a
divalent spacer between the cyclic acetal of the cyclic benzylidene
acetal group and the polyethylene glycol chain, and Z.sup.3 is a
divalent spacer between X.sup.1 and the polyethylene glycol chain.
These are composed of covalent bonds, are not particularly limited
as long as they are more stable to acid hydrolysis than the cyclic
benzylidene acetal group, and are preferably an ether bond, an
ester bond, a carbonate bond, a urethane bond, an amide bond, a
secondary amino group, an alkylene group containing any of these
bonds and group, a single bond or an alkylene group. The number of
carbon atoms of the alkylene group is preferably from 1 to 24. By
way of illustration and without limitation, preferred examples of
the alkylene group include structures such as (z1). Preferred
examples of the alkylene group having an ether bond include
structures such as (z2) or (z3). Preferred examples of the alkylene
group having an ester bond include structures such as (z4).
Preferred examples of the alkylene group having a carbonate bond
include structures such as (z5). Preferred examples of the alkylene
group having a urethane bond include structures such as (z6).
Preferred examples of the alkylene group having an amide bond
include structures such as (z7). Preferred examples of the alkylene
group having a secondary amino group include structures such as
(z8). In a preferred embodiment, p and q are each independently an
integer of 1 to 12. However, in the case where at least one of
Z.sup.1, Z.sup.2 and Z.sup.3 is an ether bond, an ester bond, a
carbonate bond, a urethane bond, an amide bond, a secondary amino
group or an alkylene group containing any of these bonds and group
and a plurality of identical structural units are connected, a
number of the structural units described above is 2 or less.
##STR00016##
[0067] P.sup.1 in formula (1) or formula (2) of the invention is a
straight-chain or branched polyethylene glycol having a number of
ethylene glycol units of 3 or more, and P.sup.2 is a straight-chain
or branched polyethylene glycol having a number of ethylene glycol
units of 3 or more. The number of the ethylene glycol units
constituting P.sup.1 or P.sup.2 is more preferably 10 or more, and
particularly preferably 20 or more. Further, the number of the
ethylene glycol units constituting P.sup.1 or P.sup.2 is more
preferably 2,000 or less, and particularly preferably 1,000 or
less.
[0068] The term "polyethylene glycol" as used in the specification
means both of polyethylene glycol having a molecular weight
distribution obtained by polymerization of ethylene oxide and a
monodispersed polyethylene glycol obtained by connecting of an
oligoethylene glycol having a single molecular weight by a coupling
reaction.
[0069] In one aspect of the invention, the biodegradable
polyethylene glycol derivative in which w in formula (1) or formula
(2) is 1 is provided.
[0070] In a preferred embodiment of the aspect, P.sup.1 in formula
(1) or formula (2) is a straight-chain polyethylene glycol having a
hydrocarbon group or a chemically reactive functional group at the
terminal thereof.
[0071] Specific examples of the straight-chain polyethylene glycol
having a hydrocarbon group at the terminal thereof for P.sup.1
include those represented by formula (3).
Y--(OCH.sub.2CH.sub.2).sub.n-- (3)
[0072] In the formula, n is a number of repeating units per
polyethylene glycol chain, and in the polyethylene glycol having a
molecular weight distribution, it is defined that n is calculated
by various theoretical calculations based on a number average
molecular weight (Mn) of the compound.
[0073] In the formula. Y is a hydrocarbon group having from 1 to 24
carbon atoms, specific examples thereof include a methyl group, an
ethyl group, a propyl group, an isopropyl group, a butyl group, a
tert-butyl group, a pentyl group, an isopentyl group, a hexyl
group, a heptyl group, a 2-ethylhexyl group, an octyl group, a
nonyl group, a decyl group, an undecyl group, a dodecyl group, a
tridecyl group, a tetradecyl group, a pentadecyl group, a hexadecyl
group, a heptadecyl group, an octadecyl group, a nonadecyl group,
an eicosyl group, a heneicosyl group, a docosyl group, a toicosyl
group, a tetracosyl group, a phenyl group, a benzyl group, a cresyl
group, a butylphenyl group, a dodecylphenyl group and a trityl
group, and Y is preferably a hydrocarbon group having from 1 to 10
carbon atoms (more preferably from 1 to 7 carbon atoms), more
preferably a methyl group or an ethyl group, and still more
preferably a methyl group.
[0074] Specific examples of the straight-chain polyethylene glycol
having a chemically reactive functional group for P.sup.1 include
those represented by formula (4).
X.sup.2--Z.sup.4--(OCH.sub.2CH.sub.2).sub.n-- (4)
[0075] In the formula, X.sup.2 is a chemically reactive functional
group different from X.sup.1, and Z.sup.4 is a divalent spacer
between the functional group X.sup.2 and the polyethylene glycol
chain. Since the biodegradable polyethylene glycol derivative has
two different chemically reactive functional groups X.sup.1 and
X.sup.2, it is possible to provide a polyethylene glycol-drug
conjugate having a target-directing property, for example, by
connecting a drug to X.sup.1 and connecting a targeting molecule to
X.sup.2.
[0076] Preferred examples of X.sup.2 include an active ester group,
an active carbonate group, an aldehyde group, an isocyanate group,
an isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group, a carboxy
group, a thiol group, a dithiopyridyl group, an .alpha.-haloacetyl
group, an alkynyl group, an allyl group, a vinyl group, an amino
group, an oxyamino group, a hydrazide group and an azide group.
[0077] More specifically, the functional group capable of forming a
covalent bond upon a reaction with an amino group of the
biofunctional molecule is an active ester group, an active
carbonate group, an aldehyde group, an isocyanate group, an
isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group or a carboxy
group, the functional group capable of forming a covalent bond upon
a reaction with a thiol group of the biofunctional molecule is an
active ester group, an active carbonate group, an aldehyde group,
an isocyanate group, an isothiocyanate group, an epoxy group, a
maleimide group, a vinyl sulfone group, an acryl group, a
sulfonyloxy group, a carboxy group, a thiol group, a dithiopyridyl
group, an .alpha.-haloacetyl group, an alkynyl group, an allyl
group or a vinyl group, the functional group capable of forming a
covalent bond upon a reaction with an aldehyde group or a carboxy
group of the biofunctional molecule is a thiol group, an amino
group, an oxyamino group or a hydrazide group, the functional group
capable of forming a covalent bond upon a reaction with an alkynyl
group of the biofunctional molecule is a thiol group or an azide
group, and the functional group capable of forming a covalent bond
upon a reaction with an azide group of the biofunctional molecule
is an alkynyl group.
[0078] In a preferred embodiment of the invention, X.sup.2 is a
group represented by group (I), group (II), group (III), group (IV)
or group (V).
Group (I): Functional group capable of forming a covalent bond upon
a reaction with an amino group of the biofunctional molecule
[0079] (a), (b), (c), (d), (e) and (f) shown below:
Group (II): Functional group capable of forming a covalent bond
upon a reaction with a thiol group of the biofunctional
molecule
[0080] (a), (b), (c), (d), (e), (f), (g), (h), (i) and (j) shown
below:
Group (III): Functional group capable of forming a covalent bond
upon a reaction with an aldehyde group or a carboxy group of the
biofunctional molecule
[0081] (g), (k), (l) and (m) shown below:
Group (IV): Functional group capable of forming a covalent bond
upon a reaction with an alkynyl group of the biofunctional
molecule
[0082] (g), (k), (l), (m) and (n) shown below:
Group (V): Functional group capable of forming a covalent bond upon
a reaction with an azide group of the biofunctional molecule
[0083] (j) shown below:
##STR00017##
[0084] In the formulae above, R.sup.7 is a hydrogen atom or a sulfo
group, specific examples of the sulfo group include sodium
sulfonate and potassium sulfonate, and R.sup.7 is preferably a
hydrogen atom. R.sup.8 and R.sup.11 are each a hydrogen atom or a
hydrocarbon group having from 1 to 5 carbon atoms, and specific
examples of the hydrocarbon group include a methyl group, an ethyl
group, a propyl group, an isopropyl group, a butyl group, a
tert-butyl group and a pentyl group. R.sup.9 is a hydrocarbon group
having from 1 to 10 carbon atoms which may contain a halogen atom,
specific examples of the hydrocarbon group include a methyl group,
an ethyl group, a propyl group, an isopropyl group, a butyl group,
a tert-butyl group, a pentyl group, an isopentyl group, a hexyl
group, a benzyl group, a 4-methylphenyl group, a trifluoromethyl
group, a 2,2,2-trifluoroethyl group, a 4-(trifluoromethoxy)phenyl
group, a vinyl group, a chloroethyl group, a bromoethyl group and
an iodoethyl group, and R.sup.9 is preferably a methyl group, a
vinyl group, a 4-methylphenyl group or a 2,2,2-trifluoroethyl
group. R.sup.10 is a halogen atom selected from the group
consisting of a chlorine atom, a bromine atom and an iodine
atom.
[0085] It is necessary that X.sup.2 is different from X.sup.1. As
to preferred examples of a combination of X.sup.1 and X.sup.2, when
X.sup.1 is an active ester group or an active carbonate group,
X.sup.2 is a group selected from a maleimide group, a vinyl sulfone
group, an .alpha.-haloacetyl group, an alkynyl group and an azide
group; when X.sup.1 is an aldehyde group, X.sup.2 is a group
selected from a maleimide group, a vinyl sulfone group, an alkynyl
group and an azide group; when X.sup.1 is a maleimide group, a
vinyl sulfone group or an .alpha.-haloacetyl group, X.sup.2 is a
group selected from an active ester group, an active carbonate
group, an alkynyl group and an azide group; when X.sup.1 is an
alkynyl group or an azide group, X.sup.2 is a group selected from a
maleimide group, a vinyl sulfone group, an .alpha.-haloacetyl
group, an active ester group, an active carbonate group, an amino
group and an oxyamino group; when X.sup.1 is an amino group or an
oxyamino group, X.sup.2 is an alkynyl group, an azide group, a
thiol group or a carboxy group; and when X.sup.1 is a thiol group.
X.sup.2 is a group selected from an amino group, an oxyamino group,
an azide group and a carboxy group. More preferably, when X.sup.1
is an active ester group or an active carbonate group, X.sup.2 is a
group selected from a maleimide group, an .alpha.-haloacetyl group,
an alkynyl group and an azide group; when X.sup.1 is an aldehyde
group, X.sup.2 is a group selected from a maleimide group, an
.alpha.-haloacetyl group, an alkynyl group and an azide group; when
X.sup.1 is a maleimide group or an .alpha.-haloacetyl group,
X.sup.2 is a group selected from an active ester group, an active
carbonate group, an alkynyl group and an azide group; when X.sup.1
is an alkynyl group or an azide group, X.sup.2 is a group selected
from a maleimide group, an .alpha.-haloacetyl group, an active
ester group, an active carbonate group, an amino group and an
oxyamino group; when X.sup.1 is an amino group or an oxyamino
group, X.sup.2 is an alkynyl group, an azide group or a thiol
group; and when X.sup.1 is a thiol group, X.sup.2 is a group
selected from an amino group, an oxyamino group and an azide
group.
[0086] Z.sup.4 is composed of covalent bonds, is not particularly
limited as long as it is more stable to acid hydrolysis than the
cyclic benzylidene acetal group, and is preferably an ether bond,
an ester bond, a carbonate bond, a urethane bond, an amide bond, a
secondary amino group, an alkylene group containing any of these
bonds and group, a single bond or an alkylene group. The number of
carbon atoms of the alkylene group is preferably from 1 to 24. By
way of illustration and without limitation, preferred examples of
the alkylene group include structures such as (z1) shown below.
Preferred examples of the alkylene group having an ether bond
include structures such as (z2) or (z3) shown below. Preferred
examples of the alkylene group having an ester bond include
structures such as (z4) shown below. Preferred examples of the
alkylene group having a carbonate bond include structures such as
(z5) shown below. Preferred examples of the alkylene group having a
urethane bond include structures such as (z6) shown below.
Preferred examples of the alkylene group having an amide bond
include structures such as (z7) shown below. Preferred examples of
the alkylene group having a secondary amino group include
structures such as (z8) shown below. In a preferred embodiment, p
and q are each independently an integer of 1 to 12. However, in the
case where Z.sup.3 is an ether bond, an ester bond, a carbonate
bond, a urethane bond, an amide bond, a secondary amino group or an
alkylene group containing any of these bonds and group and a
plurality of identical structural units are connected, a number of
the structural units described above is 2 or less.
##STR00018##
[0087] In another preferred embodiment of the aspect, P.sup.1 in
formula (1) or formula (2) is a branched polyethylene glycol having
a hydrocarbon group or a chemically reactive functional group at
the terminal thereof.
[0088] Specific examples of the branched polyethylene glycol having
a hydrocarbon group at the terminal thereof for P.sup.1 include
those represented by formula (5).
##STR00019##
[0089] In the formula, Y is a hydrocarbon group having from 1 to 24
carbon atoms as described above, and a is 0 or 2.
[0090] In the case where a is 0, two polyethylene glycol chains are
present and, in the case where a is 2, four polyethylene glycol
chains are present. In general, in the chemical modification of a
bio-related substance with polyethylene glycol, when connecting
points to the polyethylene glycol are introduced more than
necessary, the active sites of the bio-related substance are
destroyed to reduce its function so that an attempt to increase the
effect by increasing a molecular weight of the polyethylene glycol
has been performed. However, the viscosity increases with the
increase in the molecular weight and hence, for example, handling
as an aqueous solution preparation, for example, an injection
preparation becomes difficult. Since the polyethylene glycol
derivative has a branched structure, it shows low viscosity in
comparison with a straight-chain polyethylene glycol derivative
having the same molecular weight, and thus it is useful in
application, for example, the aqueous solution preparation.
[0091] Specific examples of the branched polyethylene glycol having
a chemically reactive functional group at the terminal thereof for
P.sup.1 include those represented by formula (6).
##STR00020##
[0092] In the formula, X.sup.2 is a chemically reactive functional
group different from X.sup.1 as described above, Z.sup.4 is a
divalent spacer as described above, and a is 0 or 2.
[0093] The polyethylene glycol derivative in which P.sup.1 is
represented by formula (6) has one X.sup.1 and two or four X.sup.2
in the case where v in formula (1) or formula (2) is 1 and, for
example, when a drug is connected to X.sup.1 and a targeting
molecule is connected to X.sup.2, high target-directing performance
can be obtained.
[0094] In another aspect of the invention, the biodegradable
polyethylene glycol derivative in which w in formula (1) or formula
(2) is from 2 to 8 is provided.
[0095] In a preferred embodiment of the aspect, P.sup.1 in formula
(1) or formula (2) is represented by formula (7).
##STR00021##
[0096] In the formula, X.sup.2 is a chemically reactive functional
group different from X.sup.1 as described above, Z.sup.4 is a
divalent spacer as described above, and a is 0 or 2.
[0097] In the antibody-drug conjugate (ADC)-related field, in order
to increase drug transportation efficiency, it is preferred to
connect a plurality of drugs to an antibody, but when a plurality
of connecting points are introduced into the antibody, a problem
arises in that the affinity to an antigen is decreased. The
polyethylene glycol derivative in which P.sup.1 is represented by
formula (7) has two or four X.sup.1 and one X.sup.2 in the case
where v in formula (1) or formula (2) is 1 and, for example, when
an anticancer agent is connected to X.sup.1 and an antibody is
connected to X.sup.2 in ADC targeting cancer, it is possible to
improve the transportation efficiency of the anticancer agent
without increasing the connecting points to the antibody.
[0098] In another preferred embodiment of the aspect, P.sup.1 in
formula (1) or formula (2) is polyethylene glycol having the number
of terminals of 2 to 8, all the terminals of the polyethylene
glycol constituting P.sup.1 are each connected to Z.sup.1 in
formula (1) or Z.sup.2 in formula (2), and w is equal to the number
of terminals of the polyethylene glycol.
[0099] In specific examples of the embodiment, P.sup.1 in formula
(1) or formula (2) is selected from the group consisting of formula
(r), formula (s), formula (t), formula (u) and formula (v), w is 2
in the case where P.sup.1 is represented by formula (r), w is 3 in
the case where P.sup.1 is represented by formula (s), w is 4 in the
case where P.sup.1 is represented by formula (t), w is 4 in the
case where P.sup.1 is represented by formula (u), and w is 8 in the
case where P.sup.1 is represented by formula (v).
##STR00022##
[0100] A preferred range of n in formula (3), formula (4) or
formula (r) of the invention is an integer of 3 to 2,000, more
preferably an integer of 20 to 1,000, and still more preferably an
integer of 40 to 500. Further, a preferred range of n in formula
(5), formula (6), formula (7), formula (s), formula (t), formula
(u) and formula (v) is preferably an integer of 3 to 1,000, more
preferably an integer of 10 to 500, and still more preferably an
integer of 20 to 250.
[0101] In one aspect of the invention, P.sup.2 in formula (1) or
formula (2) is represented by formula (8). Here, v in formula (1)
or formula (2) is 1.
--(OCH.sub.2CH.sub.2).sub.m-- (8)
[0102] In the formula, m is a number of repeating units per
polyethylene glycol chain, and in a polyethylene glycol having a
molecular weight distribution, it is defined that m is calculated
by various theoretical calculations based on a number average
molecular weight (Mn) of the compound.
[0103] In another aspect of the invention, P.sup.2 in formula (1)
or formula (2) is represented by formula (9).
##STR00023##
[0104] In the formula, b is 0 or 2. Here, v in formula (1) or
formula (2) is b+2.
[0105] A preferred range of m in formula (8) of the invention is an
integer of 3 to 2,000, more preferably an integer of 20 to 1,000,
and still more preferably an integer of 40 to 500. Further, a
preferred range of m in formula (9) is preferably an integer of 3
to 1,000, more preferably an integer of 10 to 500, and still more
preferably an integer of 20 to 250.
[0106] The biodegradable polyethylene glycol derivative of the
invention can be synthesized by linking a polyethylene glycol
intermediate composed of P.sup.2 to a polyethylene glycol
intermediate composed of P.sup.1 through a cyclic benzylidene
acetal linker having substituent(s). The bond generated by the
linking is determined by a combination of the functional groups
used in the reaction, and is the ether bond, the ester bond, the
carbonate bond, the urethane bond, the amide bond, the secondary
amino group, the alkylene group containing any of these bonds and
group, the single bond or the alkylene group contained in the
divalent spacer Z.sup.1 and Z.sup.2 described above. In the
biodegradable polyethylene glycol derivative synthesized, the
terminal functional group is chemically converted, if desired. As
to the reaction used for the functional group conversion, a
conventionally known method can be used, but it is necessary to
appropriately select conditions which do not decompose the cyclic
benzylidene acetal group of formula (1) or formula (2) and the
bonds contained in the divalent spacers Z.sup.1, Z.sup.2, Z.sup.3
and Z.sup.4 described above. In addition, in the synthesis of the
biodegradable polyethylene glycol derivative, the cyclic
benzylidene acetal linker compound for introducing the cyclic
benzylidene acetal linker either may be connected to the
polyethylene glycol intermediate composed of P.sup.1 and then
connected to the polyethylene glycol intermediate composed of
P.sup.2 or may be connected to the polyethylene glycol intermediate
composed of P.sup.2 and then connected to the polyethylene glycol
intermediate composed of P.sup.1. As a typical example of the
synthesis of the biodegradable polyethylene glycol derivative, the
steps described below are exemplified. A synthesis method of the
biodegradable polyethylene glycol derivative represented by formula
(1) is described herein as the typical example.
(A) Cyclic Benzylidene Acetal Linker Compound
##STR00024##
[0107] in the formula, R.sup.1 is a hydrogen atom or a hydrocarbon
group; and R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are each
independently an electron-withdrawing or electron-donating
substituent or a hydrogen atom.
##STR00025##
[0108] A carbonyl compound of formula (17) having a hydroxy group
which is a chemically reactive functional group is allowed to react
with a 1,2-diol derivative of formula (18) having a phthalimide
group in which an amino group is protected with a phthaloyl group
in an aprotic solvent, for example, toluene, benzene, xylene,
acetonitrile, ethyl acetate, diethyl ether, tert-butyl methyl
ether, tetrahydrofuran, chloroform, dichloromethane, dimethyl
sulfoxide, dimethylformamide or dimethylacetamide or with no
solvent in the presence of an acid catalyst to obtain a compound of
formula (19) shown below having a cyclic benzylidene acetal group.
The resulting compound may be purified by extraction,
recrystallization, adsorbent treatment, column chromatography or
the like. In place of the carbonyl compound, it is possible to use
a corresponding acetal derivative of a lower alcohol. The lower
alcohol is preferably an alcohol having from 1 to 5 carbon atoms,
and more preferably methanol or ethanol. The acid catalyst may be
either an organic acid or an inorganic acid and is not particularly
limited, and specific examples thereof include p-toluenesulfonic
acid, pyridinium p-toluenesulfonate, methanesulfonic acid,
10-camphorsulfonic acid, hydrogen chloride, iodine, ammonium
chloride, oxalic acid, boron trifluoride-diethyl ether complex and
the like.
##STR00026##
[0109] The "protective group" as referred to herein is a component
which prevents or blocks a reaction of a specific chemically
reactive functional group in the molecule under certain reaction
conditions. The protective group varies depending on the kind of
the chemically reactive functional group to be protected, the
conditions to be used and the presence of the other functional
group or protective group in the molecule. Specific examples of the
protective group can be found in many general books and are
described, for example, in "Wuts, P. G M.; Greene, T. W.,
Protective Groups in Organic Synthesis, 4th ed.;
Wiley-Interscience: New York, 2007". Moreover, the functional group
protected by the protective group can be reproduce the original
functional group by deprotection using reaction conditions suitable
for each of the protective groups, that is, causing a chemical
reaction. Therefore, in the specification, a functional group which
is protected by a protective group and is capable of being
deprotected by various reactions is included in the "chemically
reactive functional group". The typical deprotection conditions of
the protective group are described in the literature described
above.
[0110] As the chemically reactive functional group in the compound
of formula (17), a functional group other than the hydroxy group
can also be used. Specific examples thereof include a hydroxyalkyl
group, an amino group, an aminoalkyl group, a carboxy group and a
carboxyalkyl group. Also, the functional group described above may
be protected by a protective group which is stable in the acidic
conditions of the acetalization reaction and can be deprotected
under reaction conditions other than catalytic reduction by which
the cyclic benzylidene acetal group is decomposed. As to preferred
combinations of the functional group to be protected and the
protective group, when the functional group to be protected is a
hydroxy group or a hydroxyalkyl group, for example, a silyl
protective group and an acyl protective group are exemplified, and
specific examples thereof include a tert-butyldiphenylsilyl group,
a tert-butyldimethylsilyl group, a triisopropylsilyl group, an
acetyl group and a pivaloyl group. When the functional group to be
protected is an amino group or an aminoalkyl group, for example, an
acyl protective group and a carbamate protective group are
exemplified, and specific examples thereof include a
trifluoroacetyl group, a 9-fluorenylmethyloxycarbonyl group and a
2-(trimethylsilyl)ethyloxycarbonyl group. When the functional group
to be protected is a carboxy group or a carboxyalkyl group, for
example, an alkyl ester protective group and a silyl ester
protective group are exemplified, and specific examples thereof
include a methyl group, a 9-fluorenylmethyl group and a
tert-butyldimethylsilyl group. The kinds and the typical
deprotection conditions of the specific protective groups are
described in the literature described above, and the reaction
conditions suitable for each of the protective groups are selected
and the deprotection can be performed before the reaction with the
hydrophilic polymer intermediate.
[0111] Moreover, as the chemically reactive functional group
excepting the 1,2-diol moiety in the compound of formula (18), a
functional group other than the phthalimide group can also be used.
In the case where the chemically reactive functional group is a
functional group which is protected by a protective group, it is
necessary that the protective group is stable in the acidic
conditions of the acetalization reaction and can be deprotected
under reaction conditions other than catalytic reduction by which
the benzylidene acetal group is decomposed.
[0112] As to preferred combinations of the functional group to be
protected and the protective group, when the functional group to be
protected is an amino group, for example, an acyl protective group
and a carbamate protective group are exemplified, and specific
examples thereof include a trifluoroacetyl group, a
9-fluorenylmethyloxycarbonyl group and a
2-(trimethylsilyl)ethyloxycarbonyl group. When the functional group
to be protected is a hydroxy group, for example, a silyl protective
group and an acyl protective group are exemplified, and specific
examples thereof include a tert-butyldiphenylsilyl group, a
tert-butyldimethylsilyl group, a triisopropylsilyl group, an acetyl
group and a pivaloyl group. When the functional group to be
protected is a carboxy group, for example, an alkyl ester
protective group and a silyl ester protective group are
exemplified, and specific examples thereof include a methyl group,
a 9-fluorenylmethyl group and a tert-butyldimethylsilyl group. When
the functional group to be protected is a sulfanyl group, for
example, a thioether protective group, a thiocarbonate protective
group and a disulfide protective group are exemplified, and
specific examples thereof include an S-2,4-dinotrophenyl group, an
S-9-tluorenylmethyloxycarbonyl group and an S-tert-butyldisulfide
group. The typical deprotection conditions of the protective group
are described in the literature described above, and the reaction
conditions suitable for each of the protective groups are selected.
However, in the case where the chemically reactive functional group
is a functional group which does not inhibit the acetalization
reaction even when it is not protected by a protective group, it is
not necessary to use a protective group.
(B) Polyethylene Glycol Intermediate Composed of P.sup.1
[0113] Ethylene oxide is polymerized in an amount of 3 to 2,000
molar equivalents to methanol, which is an initiator, in toluene or
with no solvent under alkaline conditions, for example, metallic
sodium, metallic potassium, sodium hydride or potassium hydride to
obtain polyethylene glycol of formula (20). The initiator is
preferably an alcohol having a hydrocarbon group having from 1 to
24 carbon atoms, and specifically includes, for example, methanol,
ethanol, propanol, isopropanol, butanol, tert-butanol, phenol and
benzyl alcohol. Since the polyethylene glycol has a hydroxy group
which is a chemically reactive functional group, it can be used as
it is in a reaction with a cyclic benzylidene acetal linker
compound.
CH.sub.3--(OCH.sub.2CH.sub.2).sub.n--OH (20)
[0114] The polyethylene glycol of formula (20) is allowed to react
with methanesulfonyl chloride in an aprotic solvent, for example,
toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl
ether, tert-butyl methyl ether, tetrahydrofuran, chloroform,
dichloromethane, dimethyl sulfoxide, dimethylformamide or
dimethylacetamide or with no solvent in the presence of an organic
base, for example, triethylamine, N-methylmorpholine, pyridine or
4-dimethylaminopyridine or an inorganic base, for example, sodium
carbonate, sodium hydrogen carbonate, sodium acetate or potassium
carbonate to obtain a polyethylene glycol intermediate of formula
(21). The organic base and inorganic base may not be used. The use
ratio of the organic base or the inorganic base is not particularly
limited, and is preferably equimolar or more to the hydroxy group
of the polyethylene glycol of formula (20). Also, it is possible to
use the organic base as a solvent. The resulting compound may be
purified by a purification means, for example, extraction,
recrystallization, adsorbent treatment, reprecipitation, column
chromatography or supercritical extraction.
##STR00027##
[0115] As the chemically reactive functional group in the
polyethylene glycol intermediate of formula (21), other functional
groups can be also used. Preferred examples of the chemically
reactive functional group are functional groups wherein the bond
generated by the reaction of the polyethylene glycol intermediate
with the cyclic benzylidene acetal linker compound described above
becomes the ether bond, the ester bond, the carbonate bond, the
urethane bond, the amide bond, the secondary amino group, the
alkylene group containing any of these bonds and group, the single
bond or the alkylene group contained in the divalent spacer Z.sup.1
of formula (1), and specifically include, for example, a halogen
atom, an active ester, an active carbonate, an aldehyde group, an
amino group, a hydroxy group and a carboxy group.
(C) Reaction Between Cyclic Benzylidene Acetal Linker Compound and
Polyethylene Glycol Intermediate Composed of P.sup.1
[0116] The benzylidene acetal linker compound of formula (19) and
the polyethylene glycol intermediate of formula (21) are subjected
to a reaction in an aprotic solvent, for example, toluene, benzene,
xylene, acetonitrile, ethyl acetate, diethyl ether, tert-butyl
methyl ether, tetrahydrofuran, chloroform, dichloromethane,
dimethyl sulfoxide, dimethylformamide or dimethylacetamide or with
no solvent in the presence of an organic base, for example,
triethylamine, N-methylmorpholine, potassium tert-butoxide or
sodium hexamethyldisilazane or an inorganic base, for example,
potassium carbonate, potassium hydroxide or sodium hydride to
obtain a compound of formula (22). The use ratio of the organic
base or the inorganic base is not particularly limited, and is
preferably equimolar or more to the chemically reactive functional
group of the polyethylene glycol intermediate of formula (21).
Also, it is possible to use the organic base as a solvent. The
resulting compound may be purified by the purification means
described above.
##STR00028##
[0117] The chemically reactive functional group of the cyclic
benzylidene acetal linker compound may be subjected to functional
group conversion before the reaction with the polyethylene glycol
intermediate. The reaction conditions for the reaction between the
cyclic benzylidene acetal linker compound and the polyethylene
glycol intermediate are determined depending on the combination of
the chemically reactive functional group of the cyclic benzylidene
acetal linker compound and the chemically reactive functional group
of the polyethylene glycol intermediate and a conventionally known
method can be used. However, it is necessary to appropriately
select conditions which do not decompose the bonds contained in the
cyclic benzylidene acetal group and the divalent spacers Z.sup.1
and Z.sup.2 described above of formula (1) or formula (2).
[0118] The compound of formula (22) is treated by using a basic
organic compound, for example, ethylenediamine, methyl hydrazine or
methylamine or a basic inorganic compound, for example, hydrazine,
hydroxylamine or sodium hydroxide in a protic solvent, for example,
water, methanol or ethanol, in an aprotic solvent, for example,
acetonitrile, tetrahydrofuran, dimethyl sulfoxide,
dimethylformamide or dimethylacetamide or with no solvent to obtain
a compound of formula (23) in which the phthalimide group is
deprotected and converted into an amino group. The use ratio of the
basic compound is not particularly limited, and is preferably
equimolar or more to the chemically reactive functional group of
the compound of formula (22). Also, it is possible to use the basic
compound as a solvent. The resulting compound may be purified by
the purification means described above.
##STR00029##
(D) Polyethylene Glycol Intermediate Composed of P.sup.2
[0119] The polyethylene glycol intermediate composed of P.sup.2 has
chemically reactive functional groups at at least two terminals of
polyethylene glycol, and preferred examples of the chemically
reactive functional group include an active ester group, an active
carbonate group, an aldehyde group, an isocyanate group, an
isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group, a carboxy
group, a thiol group, a dithiopyridyl group, an .alpha.-haloacetyl
group, an alkynyl group, an allyl group, a vinyl group, an amino
group, an oxyamino group, a hydrazide group, an azide group and a
hydroxy group. More specifically, the functional group capable of
forming a covalent bond upon a reaction with an amino group of the
cyclic acetal linker is an active ester group, an active carbonate
group, an aldehyde group, an isocyanate group, an isothiocyanate
group, an epoxy group, a maleimide group, a vinyl sulfone group, an
acryl group, a sulfonyloxy group or a carboxy group, the functional
group capable of forming a covalent bond upon a reaction with a
thiol group of the cyclic acetal linker is an active ester group,
an active carbonate group, an aldehyde group, an isocyanate group,
an isothiocyanate group, an epoxy group, a maleimide group, a vinyl
sulfone group, an acryl group, a sulfonyloxy group, a carboxy
group, a thiol group, a dithiopyridyl group, an .alpha.-haloacetyl
group, an alkynyl group, an allyl group or a vinyl group, the
functional group capable of forming a covalent bond upon a reaction
with an aldehyde group or a carboxy group of the cyclic acetal
linker is a thiol group, an amino group, an oxyamino group, a
hydrazide group or a hydroxy group, the functional group capable of
forming a covalent bond upon a reaction with an alkynyl group of
the cyclic acetal linker is a thiol group or an azide group, and
the functional group capable of forming a covalent bond upon a
reaction with an azide group of the cyclic acetal linker is an
alkynyl group.
[0120] The chemically reactive functional groups in the
polyethylene glycol intermediate composed of P.sup.2 may be the
same or different, and a combination of two different functional
groups is preferred.
[0121] As to preferred examples of the combination of two different
functional groups, when one is an active ester group or an active
carbonate group, the other is a group selected from a maleimide
group, a vinyl sulfone group, an .alpha.-haloacetyl group, an
alkynyl group and an azide group, when one is an aldehyde group,
the other is a group selected from a maleimide group, a vinyl
sulfone group, an alkynyl group and an azide group, when one is a
maleimide group, a vinyl sulfone group or an .alpha.-haloacetyl
group, the other is a group selected from an active ester group, an
active carbonate group, an alkynyl group and an azide group, when
one is an alkynyl group or an azide group, the other is a group
selected from a maleimide group, a vinyl sulfone group, an
.alpha.-haloacetyl group, an active ester group, an active
carbonate group, an amino group, an oxyamino group and a hydroxy
group, when one is an amino group or an oxyamino group, the other
is an alkynyl group, an azide group, a thiol group, a hydroxy group
or a carboxy group, and when one is a thiol group or a hydroxy
group, the other is a group selected from an amino group, an
oxyamino group, an azide group and a carboxy group. More
preferably, when one is an active ester group or an active
carbonate group, the other is a group selected from a maleimide
group, an .alpha.-haloacetyl group, an alkynyl group and an azide
group, when one is an aldehyde group, the other is a group selected
from a maleimide group, an .alpha.-haloacetyl group, an alkynyl
group and an azide group, when one is a maleimide group or an
.alpha.-haloacetyl group, the other is a group selected from an
active ester group, an active carbonate group, an alkynyl group and
an azide group, when one is an alkynyl group or an azide group, the
other is a group selected from a maleimide group, an
.alpha.-haloacetyl group, an active ester group, an active
carbonate group, an amino group, an oxyamino group or a hydroxy
group, when one is an amino group or an oxyamino group, the other
is an alkynyl group, an azide group, a hydroxy group or a thiol
group, and when one is a thiol group or a hydroxy group, the other
is a group selected from an amino group, an oxyamino group and an
azide group.
[0122] Further, of the chemically reactive functional groups in the
polyethylene glycol intermediate composed of P.sup.2, the
functional group other than the functional groups reacted with the
cyclic acetal linker may be protected with a protective group which
is stable under the reaction conditions of the reaction with the
cyclic acetal linker and can be deprotected under reaction
conditions other than catalytic reduction by which the cyclic
benzylidene acetal group is decomposed. As to preferred
combinations of the functional group to be protected and the
protective group, when the functional group to be protected is an
amino group, for example, an acyl protective group and a carbamate
protective group are exemplified, and specific examples thereof
include a trifluoroacetyl group, a phthalimide group, a
9-fluorenylmethyloxycarbonyl group and a
2-(trimethylsilyl)ethyloxycarbonyl group. When the functional group
to be protected is a hydroxy group, for example, a silyl protective
group and an acyl protective group are exemplified, and specific
examples thereof include a tert-butyldiphenylsilyl group, a
tert-butyldimethylsilyl group, a triisopropylsilyl group, an acetyl
group and a pivaloyl group. When the functional group to be
protected is a carboxy group, for example, an alkyl ester
protective group and a silyl ester protective group are
exemplified, and specific examples thereof include a methyl group,
a 9-fluorenylmethyl group and a tert-butyldimethylsilyl group. When
the functional group to be protected is a sulfanyl group, for
example, a thioether protective group, a thiocarbonate protective
group and a disulfide protective group are exemplified, and
specific examples thereof include an S-2,4-dinotrophenyl group, an
S-9-fluorenylmethyloxycarbonyl group and an S-tert-butyldisulfide
group. The typical deprotection conditions of the protective group
are described in the literature described above, and the reaction
conditions suitable for each of the protective groups are selected.
However, in the case where the chemically reactive functional group
is a functional group which does not inhibit the reaction with the
cyclic acetal linker even when it is not protected by a protective
group, it is not necessary to use a protective group.
[0123] The description will be made here using the compound of
formula (24) having an amino group protected with a trifluoroacetyl
group at one terminal of a straight-chain polyethylene glycol and
an N-succinimidylcarbonate, which is an active carbonate group, at
the other terminal thereof. Preferred examples of the polyethylene
glycol having the combination of two different functional groups
are described, for example, in NOF Corporation (Tokyo, Japan; see
www.nof.co.jp/english: Catalogue Ver. 15), but it is not limited
thereto.
##STR00030##
(E) Reaction Between Polyethylene Glycol Intermediate Composed of
P.sup.1 Having Cyclic Benzylidene Acetal Linker and Polyethylene
Glycol Intermediate Composed of P.sup.2
[0124] The compound of formula (23) is allowed to react with the
compound of formula (24) in an aprotic solvent, for example,
toluene, benzene, xylene, acetonitrile, ethyl acetate, diethyl
ether, tert-butyl methyl ether, tetrahydrofuran, chloroform,
dichloromethane, dimethyl sulfoxide, dimethylformamide or
dimethylacetamide or with no solvent in the presence of an organic
base, for example, triethylamine, N-methylmorpholine, pyridine or
4-dimethylaminopyridine or an inorganic base, for example, sodium
carbonate, sodium hydrogen carbonate, sodium acetate or potassium
carbonate to obtain a compound of formula (25), which is a
biodegradable polyethylene glycol derivative having a cyclic
benzylidene acetal linker. The organic base or the inorganic base
may not be used. The use ratio of the organic base or the inorganic
base is not particularly limited, and is preferably equimolar or
more to the chemically reactive functional group of the compound of
formula (23). Also, it is possible to use the organic base as a
solvent.
##STR00031##
[0125] The reaction conditions of the reaction between the
polyethylene glycol intermediate composed of P.sup.1 and the
polyethylene glycol intermediate composed of P.sup.2 are determined
depending on the combination of the chemically reactive functional
group of the polyethylene glycol intermediate composed of P.sup.1
and the chemically reactive functional group of the polyethylene
glycol intermediate composed of P.sup.2 and a conventionally known
method can be used. However, it is necessary to appropriately
select conditions which do not decompose the bonds contained in the
cyclic benzylidene acetal group and the divalent spacers Z.sup.1,
Z.sup.2 and Z.sup.3 described above of formula (1) or formula
(2).
[0126] The resulting compound may be purified by a purification
means, for example, extraction, recrystallization, adsorbent
treatment, reprecipitation, column chromatography or supercritical
extraction.
[0127] As the adsorbent in the case of performing purification by
the adsorbent treatment, an inorganic adsorbent composed of an
oxide containing at least one of aluminum and silicon.
Specifically, it includes an oxide containing either one of
aluminum and silicon or both of them. More specifically, it
includes aluminum oxide, silicon dioxide, a complex oxide of
aluminum oxide and silicon dioxide, a complex oxide of aluminum
oxide and other metal, and a complex oxide of silicon dioxide and
other metal. The other metal includes sodium, magnesium and
potassium.
[0128] In the adsorption purification described above, in order to
remove impurities having an acidic functional group, an adsorbent
having an acidic substance adsorption ability is preferred, and
specific examples thereof include Kyoward 300
(2.5MgO.Al.sub.2O.sub.3.0.7CO.sub.3.nH.sub.2O), Kyoward 500
(Mg.sub.6Al.sub.2(OH).sub.16(CO.sub.3).4H.sub.2O) and Kyoward 1000
(Mg.sub.4.5Al.sub.2(OH).sub.13(CO.sub.3).3.5H.sub.2O) of Kyoward
series of Kyowa Chemical Industry Co., Ltd. The adsorbents may be
used individually or in combination.
[0129] Further, in the adsorption purification described above, in
order to remove impurities having a basic functional group, an
adsorbent having a basic substance adsorption ability is preferred,
and specific examples thereof include an adsorbent having a basic
substance adsorption ability, for example, Kyoward 600
(MgO.3SiO.sub.2.nH.sub.2O), Kyoward 700
(Al.sub.2O.sub.3.9SiO.sub.2.nH.sub.2O) or Kyoward 200B
(Al.sub.2O.sub.3.2.0H.sub.2O), preferably Kyoward 700
(Al.sub.2O.sub.3.9SiO.sub.2.nH.sub.2O) or Kyoward 200B
(Al.sub.2O.sub.32.0H.sub.2O). The adsorbents may be used
individually or in combination with other adsorbents.
[0130] Furthermore, in the adsorption purification described above,
in order to remove a neutralized salt, an adsorbent having a high
salt adsorption ability is preferred, and specific examples thereof
include Kyoward 2000 (4.5MgO.Al.sub.2O.sub.3) and Kyoward 200B
(Al.sub.2O.sub.3.2.0H.sub.2O). The adsorbents may be used
individually or in combination.
(F) Terminal Functional Group Conversion of Biodegradable
Polyethylene Glycol Derivative Having Cyclic Benzylidene Acetal
Linker
[0131] The compound of formula (25) is treated by using a basic
organic compound, for example, ethylenediamine, methyl hydrazine or
methylamine or a basic inorganic compound, for example, hydrazine,
hydroxylamine, potassium carbonate or sodium hydroxide in a protic
solvent, for example, water, methanol or ethanol, in an aprotic
solvent, for example, acetonitrile, tetrahydrofuran, dimethyl
sulfoxide, dimethylformamide or dimethylacetamide or with no
solvent to obtain a compound of formula (26) in which the
trifluoroacetyl group is deprotected and converted into an amino
group. The use ratio of the basic compound is not particularly
limited, and is preferably equimolar or more to the chemically
reactive functional group of the compound of formula (25). Also, it
is possible to use the basic compound as a solvent. The resulting
compound may be purified by the purification means described
above.
##STR00032##
[0132] Further, the compound of formula (26) is allowed to react
with N-succinimidyl 3-maleimidopropionate in an aprotic solvent,
for example, toluene, benzene, xylene, acetonitrile, ethyl acetate,
diethyl ether, tert-butyl methyl ether, tetrahydrofuran,
chloroform, dichloromethane, dimethyl sulfoxide, dimethylformamide
or dimethylacetamide or with no solvent in the presence of an
organic base, for example, triethylamine, N-methylmorpholine,
pyridine or 4-dimethylaminopyridine or an inorganic base, for
example, sodium carbonate, sodium hydrogen carbonate, sodium
acetate or potassium carbonate to obtain a compound of formula (27)
in which a maleimide group is introduced into the terminal. The
organic base and inorganic base may not be used. The use ratio of
the organic base or the inorganic base is not particularly limited,
and is preferably equimolar or more to the chemically reactive
functional group of the compound of formula (26). Also, it is
possible to use the organic base as a solvent. The resulting
compound may be purified by the purification means described
above.
##STR00033##
[0133] For the terminal functional group conversion of a
biodegradable polyethylene glycol derivative having a cyclic
benzylidene acetal linker, a conventionally known method can be
used, but it is necessary to appropriately select conditions which
do not decompose the bonds contained in the cyclic benzylidene
acetal group and the divalent spacers Z.sup.1, Z.sup.2 and Z.sup.3
described above of formula (1) or formula (2).
[0134] In formula (1) and formula (2), although the direction of
connection of the cyclic benzylidene acetal linker with respect to
P.sup.1 and P.sup.2 is opposite, the cyclic benzylidene acetal
linker compound for introducing the cyclic benzylidene acetal
linker either may be connected to the polyethylene glycol
intermediate composed of P.sup.1 and then connected to the
polyethylene glycol intermediate composed of P.sup.2 or may be
connected to the polyethylene glycol intermediate composed of
P.sup.2 and then connected to the polyethylene glycol intermediate
composed of P.sup.1, and the compound represented by formula (1)
and the compound represented by formula (2) can be synthesized
according to essentially the same technique. In addition, synthesis
examples of the compounds represented by formula (1) and formula
(2) are specifically shown in the example below, and it will be
understood by those skilled in the art that these compounds can be
synthesized according to essentially the same technique.
EXAMPLES
[0135] The invention will be described more specifically with
reference to the examples, but the invention should not be
construed as being limited thereto.
[0136] In .sup.1H-NMR analysis, JNM-ECP400 or JNM-ECA600 produced
by JEOL DATUM Ltd. was used. For the measurement, a tube of 5 mm 4
was used, and tetramethylsilane (TMS) was used as an internal
standard substance in the case where a deuterated solvent was
CDCl.sub.3, CD.sub.3CN or CD.sub.3OD, or HDO was used as a standard
in the case of D.sub.2O.
[0137] In gel permeation chromatography (GPC) analysis, there were
used SHODEX GPC SYSTEM-11 as a GPC system, SHODEX RIX8 as a
differential refractometer which was a detector, and three columns,
i.e., SHODEX KF801L, KF803L and KF804L (.phi. 8 mm.times.300 mm)
connected in series as GPC columns, and the temperature of the
column oven was set to 40.degree. C. The measurement was performed
using tetrahydrofuran as an eluent, at the flow rate of 1 mL/min,
at the sample concentration of 0.1% by weight, and in the injection
volume of 0.1 mL. The calibration curves prepared by using ethylene
glycol, diethylene glycol and triethylene glycol produced by Kanto
Chemical Co., Ltd. and Polymer Standards for GPC of polyethylene
glycol or polyethylene oxide having a molecular weight of 600 to
70,000 produced by Polymer Laboratory Co., Ltd were used. For
analysis of data, BORWIN GPC calculation program was used. Mn
represents a number average molecular weight, Mw represents a
weight average molecular weight, and a molecular weight
distribution is indicated as a calculated value of Mw/Mn.
[0138] A deuterated water buffer of MES (2-morpholinoethanesulfonic
acid) having pD of 5.5 and a deuterated water buffer of HEPES
(2-[4-(Hydroxyethyl)-1-piperazinyl]ethanesulfonic acid) having pD
of 7.4 for use in hydrolysis test were prepared by adding a 0.1M
sodium hydroxide deuterated water solution to a 0.1M MES deuterated
water solution and a 0.1M HEPES deuterated water solution,
respectively, based on the relational equation shown below
described in "Glasoe, P. K.; Long, F. A., J. Phys. Chem. 1960, 64,
188-190".
pD=Measured value by pH meter+0.40
[0139] The hydrolysis ratio of each of the compounds of formula
(35), formula (44), formula (45), formula (47) and formula (48) was
evaluated by .sup.1H-NMR and calculated according to the
calculation equation shown below by taking an integrated value of
the hydrogen of the acetal group and an integral value of the
hydrogen of the aldehyde group to be formed by hydrolysis as
I.sup.1 and I.sup.2, respectively.
Hydrolysis ratio (%)=[I.sup.2/(I.sup.1+I.sup.2)].times.100
[0140] The hydrolysis ratio of each of the compounds of formula
(41) and formula (54) was evaluated by GPC and calculated according
to the calculation equation shown below by taking a peak area of
polyethylene glycol (molecular weight: about 10,000) which was not
divided upon the hydrolysis of the linker and a peak area of
polyethylene glycol (molecular weight: about 5,000) which was
divided upon the hydrolysis of the linker as A.sup.1 and A.sup.2,
respectively.
Hydrolysis ratio (%)=[A.sup.2/(A.sup.1+A.sup.2)].times.100
[0141] The hydrolysis ratio of each of the compounds of formula
(74) and formula (76) was evaluated by GPC and calculated according
to the calculation equation shown below by taking a peak area of
polyethylene glycol (molecular weight: about 15,000) which was not
divided upon the hydrolysis of the linker, a peak area of
polyethylene glycol (molecular weight: about 10,000) which is
partially divided upon the hydrolysis of the linker and a peak area
of polyethylene glycol (molecular weight: about 5,000) which was
completely divided upon the hydrolysis of the linker as A.sup.1,
A.sup.2 and A.sup.3, respectively.
Hydrolysis ratio
(%)=[A.sup.3/(A.sup.1+A.sup.2+A.sup.3].times.100
Example 1
[0142] Into a 200 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer, a Dean-stark tube
and a condenser tube were charged 1,2,6-hexanetriol (30.0 g, 0.224
mol), acetone dimethyl acetal (25.6 g, 0.246 mol) and
p-toluenesulfonic acid monohydrate (0.426 g, 2.24 mmol), and the
reaction was performed at 80.degree. C. for 3 hours while
distilling off methanol. Triethylamine (0.453 g, 4.48 mmol) was
added thereto and the mixture was stirred for a while, diluted with
ethyl acetate, and washed with an aqueous 20% by weight sodium
chloride solution. The organic layer was dried over anhydrous
sodium sulfate, and after filtration, the solvent was distilled off
under a reduced pressure. The residue was purified by silica gel
chromatography to obtain a compound of formula (28).
[0143] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.35 (3H, s, --CH.sub.3), 1.41 (3H, s, --CH.sub.3),
1.49-1.67 (6H, m, >CHCH.sub.2HCH.sub.2CH.sub.2--), 2.07 (1H,
brs, --OH), 3.51 (1H, t, --OCH.sub.2CH<), 3.64 (2H, t,
--CH.sub.2OH), 4.04 (1H, dd, --OCH.sub.2CH<), 4.07-4.10 (1H, m,
--OCH.sub.2CH<)
##STR00034##
Example 2
[0144] Into a 500 mL four-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (28) (20.0 g, 0.115 mol), triethylamine
(23.3 g, 0.230 mol) and toluene (200 g) and the mixture was cooled
to 10.degree. C. or less. While continuing the cooling,
methanesulfonyl chloride (19.8 g, 0.173 mol) prepared in a dropping
funnel was gradually added dropwise thereto. After the completion
of the dropwise addition, the reaction was performed at 20.degree.
C. for 2 hours. Ethanol (7.97 g, 0.173 mol) was added and the
mixture was stirred for a while and filtered, and the organic layer
was washed with ion-exchanged water. The organic layer was dried
over anhydrous sodium sulfate, and after filtration, the solvent
was distilled off under a reduced pressure to obtain a compound of
formula (29).
[0145] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.35 (3H, s, --CH.sub.3), 1.40 (3H, s, --CH.sub.3),
1.44-1.83 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.01 (3H, s,
--OSO.sub.2CH.sub.3), 3.51 (1H, t, --OCH.sub.2CH<), 4.03-4.11
(2H, m, --OCH.sub.2CH<, --OCH.sub.2CH<), 4.24 (2H, t,
--CH.sub.2OSO.sub.2CH.sub.3)
##STR00035##
Example 3
[0146] Into a 500 mL four-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (29) (20.0 g, 79.3 mmol), potassium
phthalimide (17.6 g, 95.2 mmol) and dehydrated dimethylformamide
(200 g), and the reaction was performed at 60.degree. C. for 2
hours. The mixture was cooled to 10.degree. C. or less,
ion-exchanged water (400 g) was added thereto and after stirring
for a while, the mixture was extracted with a mixed solution of
ethyl acetate/hexane (60/40 in v/v). The organic layer was washed
with an aqueous 0.2% by weight potassium carbonate solution and
dried over anhydrous sodium sulfate. After filtration, the solvent
was distilled off under a reduced pressure to obtain a compound of
formula (30).
[0147] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.34 (3H, s, --CH.sub.3), 1.39 (3H, s, --CH.sub.3--),
1.44-1.75 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.50 (1H, t,
--OCH.sub.2CH<), 3.69 (2H, t, --CH.sub.2-phthalimide), 4.01-4.09
(2H, m, --OCH.sub.2CH<, --OCH.sub.2CH<), 7.71-7.85 (4H, m,
-phthalimide)
##STR00036##
Example 4
[0148] Into a 1 L four-necked flask equipped with a thermometer, a
nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (30) (15.2 g, 50.0 mmol), p-toluenesulfonic
acid monohydrate (951 mg, 5.00 mmol) and methanol (500 mL), and the
reaction was performed at room temperature for 4 hours.
Triethylamine (1.01 g, 10.0 mmol) was added thereto and after
stirring for a while, the solvent was distilled off under a reduced
pressure. The residue was dissolved in chloroform, the solution was
washed with ion-exchanged water, and the organic layer was dried
over anhydrous sodium sulfate. After filtration, the solvent was
distilled off under a reduced pressure to obtain a compound of
formula (31).
[0149] .sup.1H-NMR (CD.sub.3CN, internal standard TMS); .delta.
(ppm): 1.24-1.61 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.69
(1H, t, --OH), 2.75 (1H, d, --OH), 3.17-3.21 (1H, m,
--OCH.sub.2CH<), 3.31-3.37 (1H, m, --OCH.sub.2CH<), 3.39-3.43
(1H, m, --OCH.sub.2CH<), 3.54 (2H, t, --CH.sub.2-phthalimide),
7.67-7.75 (4H, m, -phthalimide)
##STR00037##
Example 5
[0150] Into a 300 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer, a Dean-stark tube
and a condenser tube were charged the compound of formula (31)
(3.87 g, 14.7 mmol), 4-hydroxybenzaldehyde (1.20 g, 9.83 mmol),
pyridinium p-toluenesulfonate (247 mg, 0.983 mmol) and toluene (180
g), and the reaction was performed for 4 hours while removing
by-produced water by azeotropic distillation with toluene.
Triethylamine (199 mg, 1.97 mmol) was added thereto and after
stirring for a while, the solvent was distilled off under a reduced
pressure. The residue was dissolved in chloroform, the solution was
washed in order with an aqueous 20% by weight sodium chloride
solution and ion-exchanged water, and the organic layer was dried
over anhydrous sodium sulfate. After filtration, the solvent was
distilled off under a reduced pressure to obtain a compound of
formula (32).
[0151] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.41-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.57-4.26 (5H, m, --OCH.sub.2CH<, --CH.sub.2- phthalimide), 5.71
(0.6H, s, >CH--), 5.82 (0.4H, s, >CH--), 6.79-6.82 (2H, m,
arom. H), 7.31-7.35 (2H, m, arom. H), 7.70-7.86 (4H, m,
-phthalimide)
##STR00038##
Example 6
[0152] Into a 300 mL four-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
dehydrated methanol (12.8 g, 0.400 mol), dehydrated toluene (150 g)
and metal sodium (0.3 g, 13 mmol), and the mixture was stirred at
room temperature until the metal sodium was dissolved while
bubbling nitrogen through the mixture. The solution was charged
into a 5 L autoclave and after the inside of the system was
substituted with nitrogen, temperature was raised to 100.degree. C.
After adding ethylene oxide (1.987 g, 45 mol) at 100 to 130.degree.
C. under a pressure of 1 MPa or less, the reaction was further
continued for 2 hours. After the unreacted ethylene oxide gas was
removed under a reduced pressure, the mixture was cooled to
60.degree. C., and pH was adjusted to 7.5 with an aqueous 85%
phosphoric acid solution to obtain a compound of formula (33).
[0153] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 2.68 (1H, t, OH), 3.38 (3H, s, CH.sub.3O--), 3.49-3.85
(450H, m, --(OCH.sub.2CH.sub.2).sub.n--)
[0154] GPC analysis;
[0155] Number average molecular weight (Mn): 5119, weight average
molecular weight (Mw): 5226, polydispersity (Mw/Mn): 1.021
CH.sub.3--(OCH.sub.2CH.sub.2).sub.n--OH (33) [0156] n=about 113
Example 7
[0157] Into a 500 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer, a Dean-stark tube
and a condenser tube were charged the compound of formula (33) (100
g, 20.0 mmol) and toluene (250 g), and water was removed by
azeotropic distillation with toluene. After cooling to 40.degree.
C., triethylamine (3.24 g, 32.0 mmol) was charged and
methanesulfonyl chloride (2.75 g, 24.0 mmol) prepared in a dropping
funnel was gradually added dropwise thereto. After the completion
of the dropwise addition, the reaction was performed at 40.degree.
C. for 3 hours. Ethanol (1.11 g, 24.0 mmol) was added thereto and
the mixture was stirred for a while, filtered, and diluted with
ethyl acetate (200 g). Crystallization was performed by adding
hexane (500 g), and after filtration, the crystals were dissolved
in ethyl acetate (500 g). Crystallization was again performed by
adding hexane (500 g), and after filtration, the crystals were
dried under a reduced pressure to obtain a compound of formula
(34).
[0158] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 3.08 (3H, s, --OSO.sub.2CH.sub.3), 3.38 (3H, s,
CH.sub.3O--), 3.52-3.85 (448H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--), 4.37-4.39 (2H, m,
--CH.sub.2OSO.sub.2CH.sub.3)
[0159] GPC analysis;
[0160] Number average molecular weight (Mn): 5197, weight average
molecular weight (Mw): 5306, polydispersity (Mw/Mn): 1.021
##STR00039##
Example 8
[0161] Into a 100 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer and a condenser tube
were charged the compound of formula (34) (5.00 g, 1.00 mmol), the
compound of formula (26) (551 mg, 1.50 mmol), potassium carbonate
(691 mg, 5.00 mmol) and acetonitrile (25 g), and the reaction was
performed at 80.degree. C. for 4 hours. After distilling off the
solvent under a reduce pressure, the residue was dissolved in ethyl
acetate (100 g) and the solution was filtered. Crystallization was
performed by adding hexane (100 g), and after filtration, the
crystals were dried under a reduced pressure to obtain a compound
of formula (35).
[0162] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm):
[0163] 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.38
(3H, s, CH.sub.3O--), 3.52-4.25 (455H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<,
--CH.sub.2-phthalimide), 5.72 (0.6H, s, >CH--), 5.84 (0.4H, s,
>CH--), 6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom. H),
7.70-7.86 (4H, m, -phthalimide)
[0164] GPC analysis;
[0165] Number average molecular weight (Mn): 5462, weight average
molecular weight (Mw): 5582, polydispersity (Mw/Mn): 1.022
##STR00040##
Example 9
[0166] Into a 50 mL three-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (35) (2.00 g, 0.400 mmol), methanol (7 g)
and ethylene diamine monohydrate (0.781 g, 10.0 mmol), and the
reaction was performed at 40.degree. C. for 4 hours. The mixture
was diluted with an aqueous 20% by weight sodium chloride solution,
extracted with dichloromethane, and the solvent was distilled off
under a reduced pressure. The residue was dissolved in ethyl
acetate (50 g), dried over anhydrous sodium sulfate, filtered, and
crystallized by adding hexane (50 g). After filtration, the
crystals were dried under a reduced pressure to obtain a compound
of formula (36).
[0167] .sup.1H-NMR (CD.sub.3OD, internal standard TMS); .delta.
(ppm): 1.43-1.79 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.77
(2H, t, --CH.sub.2--NH.sub.2), 3.36 (3H, s, CH.sub.3O--), 3.50-4.29
(453H, m, --(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<), 5.70
(0.6H, s, >CH--), 5.81 (0.4H, s, >CH--), 6.93-6.98 (2H, m,
arom. H), 7.33-7.41 (2H, m, arom. H)
[0168] GPC analysis;
[0169] Number average molecular weight (Mn): 5332, weight average
molecular weight (Mw): 5454, polydispersity (Mw/Mn): 1.023
##STR00041##
Example 10
##STR00042##
[0171] From the compound of formula (37) synthesized according to
the method described in JP-A-2010-248504, the tert-butyl group was
removed using hydrochloric acid to obtain a compound of formula
(38).
[0172] .sup.1H-NMR (D.sub.2O, internal standard TMS); .delta.
(ppm): 3.14 (2H, t, --CH.sub.2NH.sub.2), 3.40-4.00 (452H, m,
--(OCH.sub.2CH.sub.2).sub.m--)
H--(OCH.sub.2CH.sub.2).sub.m--NH.sub.2 (38) [0173] m=about 113
Example 11
[0174] Into a 100 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer and a condenser tube
were charged the compound of formula (38) (5.00 g, 1.00 mmol),
dichloromethane (30 g) and triethylamine (607 mg, 6.00 mmol), and
trifluoroacetic anhydride (630 mg, 3.00 mmol) was added thereto,
and the reaction was performed at 25.degree. C. for 3 hours.
Phosphate buffer having pH of 7.0 was added thereto and after
stirring for a while, the dichloromethane layer was recovered and
the solvent was distilled off under a reduced pressure. The residue
was dissolved in ethyl acetate (100 g), dried over anhydrous
magnesium sulfate, filtered, and crystallized by adding hexane (100
g). After filtration, the crystals were dried under a reduced
pressure to obtain a compound of formula (39).
[0175] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 2.58 (1H, t, --OH), 3.40-3.95 (450H, m,
--(OCH.sub.2CH.sub.2).sub.m--), 7.34 (1H, brs, --HNCOCF.sub.3)
##STR00043##
Example 12
[0176] Into a 100 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer and a condenser tube
were charged the compound of formula (39) (4.50 g, 0.900 mmol) and
dichloromethane (27 g) and N,N'-disuccinimidylcarbonate (692 mg,
2.70 mmol) and triethylamine (410 mg, 4.05 mmol) were added
thereto, and the reaction was performed at 25.degree. C. (for 4
hours. After filtration, the solvent was distilled off under a
reduced pressure. The residue was dissolved in ethyl acetate (90 g)
and crystallized by adding hexane (90 g). After filtration, the
crystals were dried under a reduced pressure to obtain a compound
of formula (40).
[0177] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 2.84 (4H, s, -succinimide), 3.40-3.95 (448H, m,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--), 4.44-4.48 (2H, m,
--CH.sub.2O--COO-succinimide), 7.34 (1H, brs, --HNCOCF.sub.3)
[0178] GPC analysis;
[0179] Number average molecular weight (Mn): 5241, weight average
molecular weight (Mw): 5356, polydispersity (Mw/Mn): 1.022
##STR00044##
Example 13
[0180] Into a 100 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer and a condenser tube
were charged the compound of formula (40) (4.00 g, 0.800 mmol), the
compound of formula (36) (4.20 g, 0.840 mmol) and toluene (24 g),
and the reaction was performed at 50.degree. C. for 2 hours. Then,
Kyoward 700 (1.2 g) was added thereto, and the adsorption treatment
was performed at 50.degree. C. for 2 hours. After filtration,
crystallization was performed by adding hexane (24 g). After
filtration, the crystals were dried under a reduced pressure to
obtain a compound of formula (41).
[0181] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.31-3.34 (2H, m, --CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--),
3.52-4.25 (903H, m, --(OCH.sub.2CH.sub.2).sub.n--,
--(OCH.sub.2CH.sub.2).sub.m--, --OCH.sub.2CH<), 5.19 (1H, brs,
--HNCOO--), 5.72 (0.6H, s, >CH--), 5.84 (0.4H, s, >CH--),
6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom. H), 7.34 (1H,
brs, --HNCOCF.sub.3)
[0182] GPC analysis;
[0183] Number average molecular weight (Mn): 10458, weight average
molecular weight (Mw): 11180, polydispersity (Mw/Mn): 1.069
##STR00045##
Example 14
[0184] Into a 100 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer and a condenser tube
were charged the compound of formula (41) (5.00 g, 0.200 mmol) and
IM aqueous potassium carbonate solution (25 g), and the reaction
was performed at 25.degree. C. for 2 hours. The mixture was diluted
with an aqueous 20% by weight sodium chloride solution, extracted
with dichloromethane, and the solvent was distilled off under a
reduced pressure. The residue was dissolved in ethyl acetate (100
g), dried over anhydrous sodium sulfate, filtered, and crystallized
by adding hexane (100 g). After filtration, the crystals were dried
under a reduced pressure to obtain a compound of formula (42).
[0185] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.86
(2H, t, --CH.sub.2--NH.sub.2), 3.31-3.34 (2H, m,
--CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--), 3.52-4.25 (901H,
m, --(OCH.sub.2CH.sub.2).sub.n--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<), 5.19
(1H, brs, --HNCOO--), 5.72 (0.6H, s, >CH--), 5.84 (0.4H, s,
>CH--), 6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom.
H)
[0186] GPC analysis;
[0187] Number average molecular weight (Mn): 10309, weight average
molecular weight (Mw): 11021, polydispersity (Mw/Mn): 1.069
##STR00046##
Example 15
[0188] Into a 50 mL three-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (42) (2.00 g, 0.200 mmol) and toluene (10
g), and N-succinimidyl 3-maleimidopropionate (63.9 mg, 0.240 mmol)
was added thereto, and the reaction was performed at 40.degree. C.
for 2 hours. After filtration, the mixture was diluted with ethyl
acetate (40 g), and crystallized by adding hexane (50 g). After
filtration, the crystals were dried under a reduced pressure to
obtain a compound of formula (43).
[0189] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.44
(2H, t, --CH.sub.2CH.sub.2-maleimide), 3.27-3.34 (4H, m,
--CH.sub.2--HNCOO--, --CH.sub.2NHCOCH.sub.2--), 3.38 (3H, s,
CH.sub.3O--), 3.52-4.25 (903H, m, --(OCH.sub.2CH.sub.2).sub.n--,
--(OCH.sub.2CH.sub.2).sub.m, --OCH.sub.2CH<,
--CH.sub.2CH.sub.2-maleimide), 5.19 (1H, brs, --HNCOO--), 5.72
(0.6H, s, >CH--), 5.84 (0.4H, s, >CH--), 6.70 (2H, s,
-maleimide), 6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom.
H)
[0190] GPC analysis;
[0191] Number average molecular weight (Mn): 10513, weight average
molecular weight (Mw): 11249, polydispersity (Mw/Mn): 1.070
##STR00047##
Example 16
[0192] A compound of formula (44) was obtained in the same manner
as in Examples 1 to 8 using 3-fluoro-4-hydroxybenzaldehyde.
[0193] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.38-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.38
(3H, s, CH.sub.3O--), 3.52-4.23 (455H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<,
--CH.sub.2-phthalimide), 5.70 (0.6H, s, >CH--), 5.82 (0.4H, s,
>CH--), 6.95-7.21 (3H, m, arom. H), 7.70-7.86 (4H, m,
-phthalimide)
[0194] GPC analysis;
[0195] Number average molecular weight (Mn): 5485, weight average
molecular weight (Mw): 5606, polydispersity (Mw/Mn): 1.022
##STR00048##
Example 17
[0196] A compound of formula (45) was obtained in the same manner
as in Examples 1 to 8 using 2-bromo-5-hydroxybenzaldehyde.
[0197] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.38-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.38
(3H, s, CH.sub.3O--), 3.52-4.23 (455H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<,
--CH.sub.2-phthalimide), 5.70 (0.6H, s, >CH--), 5.82 (0.4H, s,
>CH--), 6.95-7.21 (3H, m, arom. H), 7.70-7.86 (4H, m,
-phthalimide)
[0198] GPC analysis;
[0199] Number average molecular weight (Mn): 5548, weight average
molecular weight (Mw): 5670, polydispersity (Mw/Mn): 1.022
##STR00049##
Example 18
##STR00050##
[0201] A compound of formula (46) was synthesized in a manner
similar to Examples 1 to 4, and a compound of formula (47) was
obtained in the same manner as in Examples 5 to 8 using
3-fluoro-4-hydroxybenzaldehyde.
[0202] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.89 (2H, m, --CH.sub.2CH.sub.2-phthalimide), 3.19 (1H, m,
--OCH.sub.2CH<), 3.38 (3H, s, CH.sub.3O--), 3.52-4.41 (456H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<,
--CH.sub.2CH.sub.2CH.sub.2-phthalimide), 5.34 (0.8H, s, >CH--),
5.42 (0.2H, s, >CH--), 6.95-7.25 (3H, m, arom. H), 7.70-7.86
(4H, m, -phthalimide)
[0203] GPC analysis;
[0204] Number average molecular weight (Mn): 5498, weight average
molecular weight (Mw): 5619, polydispersity (Mw/Mn): 1.022
##STR00051##
Example 19
[0205] Using the compound of formula (46) and
2-bromo-5-hydroxybenzaldehyde, a compound of formula (48) was
obtained in the same manner as in Examples 5 to 8.
[0206] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.89 (2H, m, --CH.sub.2CH.sub.2-phthalimide), 3.19 (1H, m,
--OCH.sub.2CH<), 3.38 (3H, s, CH.sub.3O--), 3.52-4.41 (456H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --OCH.sub.2CH<,
--CH.sub.2CH.sub.2CH.sub.2-phthalimide), 5.61 (0.8H, s, >CH--),
5.68 (0.2H, s, >CH--), 6.78-7.40 (3H, m, arom. H), 7.70-7.86
(4H, m, -phthalimide)
[0207] GPC analysis;
[0208] Number average molecular weight (Mn): 5564, weight average
molecular weight (Mw): 5686, polydispersity (Mw/Mn): 1.022
##STR00052##
Example 20
[0209] The compound of formula (39) was allowed to react with
methanesulfonyl chloride in a manner similar to Example 7 to obtain
a compound of formula (49).
[0210] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 3.08 (3H, s, --OSO.sub.2CH.sub.2), 3.40-3.95 (448H, m,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--), 4.37-4.39 (2H, m,
--CH.sub.2OSO.sub.2CH.sub.3), 7.34 (1H, brs, --HNCOCF.sub.3)
[0211] GPC analysis;
[0212] Number average molecular weight (Mn): 5193, weight average
molecular weight (Mw): 5302, polydispersity (Mw/Mn): 1.021
##STR00053##
Example 21
[0213] Into a 300 mL three-necked flask equipped with a
thermometer, a nitrogen inlet tube, a stirrer, a Dean-stark tube
and a condenser tube were charged 1,2,6-hexanetriol (2.01 g, 15.0
mmol), 3-fluoro-4-hydroxybenzaldehyde (1.40 g, 10.0 mmol),
p-toluenesulfonic acid monohydrate (19.0 mg, 0.100 mmol) and
toluene (183 g), and the reaction was performed for 4 hours while
removing by-produced water by azeotropic distillation with toluene.
Triethylamine (20.2 mg, 0.200 mmol) was added thereto and after
stirring for a while, the solution was washed with an aqueous 10%
by weight sodium chloride solution, and the organic layer was dried
over anhydrous sodium sulfate. After filtration, the solvent was
distilled off under a reduced pressure to obtain a compound of
formula (50).
[0214] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.32-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.50-4.24 (5H, m, --OCH.sub.2CH<, --CH.sub.2--OH), 5.71 (0.6H,
s, >CH--), 5.82 (0.4H, s, >CH--), 6.73-7.24 (3H, m, arom.
H)
##STR00054##
Example 22
[0215] Using the compound of formula (50) and the compound of
formula (49), a compound of formula (51) was obtained in the same
manner as in Example 8.
[0216] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CH.sub.2CH.sub.2CH.sub.2--), 3.40-4.25
(455H, m, --(OCH.sub.2CH.sub.2--).sub.m--, --OCH.sub.2CH<,
--CH.sub.2--OH), 5.70 (0.6H, s, >CH--), 5.82 (0.4H, s,
>CH--), 6.95-7.21 (3H, m, arom. H), 7.34 (1H, brs,
--HNCOCF.sub.3)
[0217] GPC analysis;
[0218] Number average molecular weight (Mn): 5239, weight average
molecular weight (Mw): 5354, polydispersity (Mw/Mn): 1.022
##STR00055##
Example 23
[0219] The compound of formula (51) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12 to
obtain a compound of formula (52).
[0220] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.84
(4H, s, -succinimide), 3.40-4.25 (453H, m,
--(OCH.sub.2CH.sub.2).sub.m--, --OCH.sub.2CH<), 4.33 (2H, dd,
--CH.sub.2O--COO-succinimide), 5.70 (0.6H, s, >CH--), 5.82
(0.4H, s, >CH--), 6.95-7.21 (31H, m, arom. H), 7.34 (1H, brs,
--HNCOCF.sub.3)
[0221] GPC analysis;
[0222] Number average molecular weight (Mn): 5354, weight average
molecular weight (Mw): 5472, polydispersity (Mw/Mn): 1.022
##STR00056##
Example 24
[0223] CH.sub.3--(OCH.sub.2CH.sub.2).sub.n--NH.sub.2 (53) [0224]
n=about 113
[0225] Using the compound of formula (53) synthesized according to
the method described in JP-A-2010-248504 and the compound of
formula (52), a compound of formula (54) was obtained in the same
manner as in Example 13.
[0226] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.27-3.29 (2H, m, --CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--),
3.52-4.25 (903H, m, --(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--, --OCH.sub.2.sub.2CH<,
--HNCOO--CH.sub.2--), 5.19 (1H, brs, --HNCOO--), 5.70 (0.6H, s,
>CH--), 5.82 (0.4H, s, >CH--), 6.95-7.21 (3H, m, arom. H),
7.34 (1H, brs, --HNCOCF.sub.3)
[0227] GPC analysis;
[0228] Number average molecular weight (Mn): 10138, weight average
molecular weight (Mw): 10685, polydispersity (Mw/Mn): 1.054
##STR00057##
Example 25
[0229] The compound of formula (54) was subjected to deprotection
of the trifluoroacetyl group in the same manner as in Example 14,
followed by allowing to react with N-succinimidyl
3-maleimidopropionate in the same manner as in Example 15 to obtain
a compound of formula (55).
[0230] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.44
(2H, t, --CH.sub.2CH.sub.2-maleimide), 3.27-3.29 (4H, m,
--CH.sub.2--HNCOO--, --CH.sub.2--NHCOCH.sub.2--), 3.38 (3H, s,
CH.sub.3O--), 3.52-4.25 (903H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--, --CH.sub.2CH.sub.2-maleimide), 5.19 (1H, brs,
--HNCOO--), 5.70 (0.61, s, >CH--), 5.82 (0.4H, s, >CH--),
6.15 (1H, brs, --HNCOCH.sub.2--), 6.70 (2H, s, -maleimide),
6.95-7.21 (3H, m, arom. H)
[0231] GPC analysis;
[0232] Number average molecular weight (Mn): 10291, weight average
molecular weight (Mw): 10847, polydispersity (Mw/Mn): 1.054
##STR00058##
Example 26
##STR00059##
[0234] The compound of formula (56) synthesized according to the
method described in JP-A-2004-197077 was allowed to react with
acetic anhydride in the presence of triethylamine and
4-dimethylaminopyridine to obtain a compound of formula (57).
[0235] .sup.1H-NMR (CDCl.sub.3, internal standard TMS): .delta.
(ppm):
[0236] 2.08 (6H, s, CH.sub.3CO--), 3.40-4.00 (901H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.n--OCH<, --CH.sub.2OCH.sub.2Ph), 4.22
(4H, t, CH.sub.3CO.sub.2CH.sub.2--), 4.54 (2H, s,
--CH.sub.2OCH.sub.2Ph), 7.27-7.38 (5H, m,
--CH.sub.2OCH.sub.2Ph)
##STR00060##
Example 27
[0237] From the compound of formula (57), the benzyl group was
removed according to the method described in JP-A-2004-197077,
followed by allowing to react with methanesulfonyl chloride in a
manner similar to Example 7 to obtain a compound of formula
(58).
[0238] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 2.08 (6H, s, CH.sub.3CO--), 3.08 (3H, s,
--OSO.sub.2CH.sub.3), 3.40-4.00 (899H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.n--OCH<), 4.22 (4H, t,
CH.sub.3CO.sub.2CH.sub.2--), 4.26-4.42 (2H, m,
--CH.sub.2OSO.sub.2CH.sub.3)
##STR00061##
Example 28
[0239] Using 3-fluoro-4-hydroxybenzaldehyde and the compound of
formula (58), a compound of formula (59) was obtained in the same
manner as in Examples 1 to 5 and 8.
[0240] .sup.1H-NMR (CDCl.sub.3, internal standard TMS): .delta.
(ppm): 1.38-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.08
(6H, s, CH.sub.3CO--), 3.40-4.23 (910H, m,
--(OCH.sub.2CH.sub.2)--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.n--OCH<, --OCH.sub.2CH<,
--CH.sub.2-phthalimide, CH.sub.3CO.sub.2CH.sub.2--), 5.70 (0.6H, s,
>CH--), 5.82 (0.4H, s, >CH--), 6.95-7.21 (3H, m, arom. H),
7.70-7.86 (4H, m, -phthalimide)
[0241] GPC analysis;
[0242] Number average molecular weight (Mn): 10223, weight average
molecular weight (Mw): 10458, polydispersity (Mw/Mn): 1.023
##STR00062##
Example 29
[0243] The compound of formula (38) was allowed to react with
5-azidopentanoic anhydride and then allowed to react with
N,N'-disuccinimidylcarbonate to obtain a compound of formula
(60).
[0244] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.60-1.74 (4H, m,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 2.18 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 2.84 (4H, s,
-succinimide), 3.29 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 3.40-3.85 (448H, m,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--), 4.44-4.48 (2H, m,
--CH.sub.2O--COO-succinimide), 6.30 (1H, brs, --HNCOCH.sub.2--)
[0245] GPC analysis;
[0246] Number average molecular weight (Mn): 5532, weight average
molecular weight (Mw): 5665, polydispersity (Mw/Mn): 1.024
##STR00063##
Example 30
[0247] A compound of formula (59) was subjected to deprotection of
the phthalimide group using ethylene diamine monohydrate and to
removal of the acetyl group using an aqueous sodium hydroxide
solution, followed by allowing to react with the compound of
formula (60) to obtain a compound of formula (61).
[0248] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm):
[0249] 1.40-1.81 (10H, m, >CHCH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 2.18 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 3.29 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 3.31-3.34 (2H, m,
--CH.sub.2--HNCOO--), 3.40-4.23 (1353H, m, --(OCH.sub.2OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.n--OCH<, --OCH.sub.2CH<), 5.19 (1H,
brs, --HNCOO--), 5.70 (0.6H, s, >CH--), 5.82 (0.4H, s,
>CH--), 6.30 (1H, brs, --HNCOCH.sub.2--), 6.95-7.21 (3H, m,
{right arrow over (arom. H)})
[0250] GPC analysis:
[0251] Number average molecular weight (Mn): 14728, weight average
molecular weight (Mw): 15582, polydispersity (Mw/Mn): 1.058
##STR00064##
Example 31
[0252] The compound of formula (61) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12 to
obtain a compound of formula (62).
[0253] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm):
[0254] 1.40-1.81 (10H, m, >CHCH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH.sub.2CH.sub.2N.sub.3), 2.18 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 2.84 (8H, s,
-succinimide), 3.29 (2H, t,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2N.sub.3), 3.31-3.34 (2H, m,
--CH.sub.2--HNCOO--), 3.40-4.23 (1349H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.n--OCH<, --OCH.sub.2CH<), 4.44-4.48
(4H, m, --CH.sub.2O--COO-succinimide), 5.19 (1H, brs, --HNCOO--),
5.70 (0.6H, s, >CH--), 5.82 (0.4H, s, >CH--), 6.30 (1H, brs,
--HNCOCH.sub.2--), 6.95-7.21 (3H, m, arom. H)
[0255] GPC analysis;
[0256] Number average molecular weight (Mn): 14958, weight average
molecular weight (Mw): 15855, polydispersity (Mw/Mn): 1.060
##STR00065##
Example 32
##STR00066##
[0258] The compound of formula (63) synthesized by polymerizing
ethylene oxide to pentaerythritol was allowed to react with
methanesulfonyl chloride in the same manner as in Example 7 to
obtain a compound of formula (64).
[0259] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 3.08 (12H, s, --OSO.sub.2CH.sub.3), 3.47-3.85 (1800H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--), 4.37-4.39 (8H, m,
--CH.sub.2OSO.sub.2CH.sub.3)
##STR00067##
Example 33
[0260] Using the compound of formula (64) and the compound of
formula (50), a compound of formula (65) was obtained in the same
manner as in Example 8.
[0261] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (24H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.52-4.23 (1828H, m, --(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--OCH.sub.2CH<, --CH--OH), 5.70 (2.4H, s, >CH--), 5.82 (1.6H,
s, >CH--), 6.95-7.21 (12H, m, arom. H)
[0262] GPC analysis;
[0263] Number average molecular weight (Mn): 19078, weight average
molecular weight (Mw): 19574, polydispersity (Mw/Mn): 1.026
##STR00068##
Example 34
[0264] The compound of formula (65) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12 to
obtain a compound of formula (66).
[0265] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (24H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.84
(16H, s, -succinimide), 3.52-4.23 (1820H, m, --(OCH.sub.2CH.sub.2),
--OCH.sub.2--, --OCH.sub.2CH<), 4.33 (811, dd,
--CH.sub.2O--COO-succinimide), 5.70 (2.4H, s, >CH--), 5.82
(1.6H, s, >CH--), 6.95-7.21 (12H, m, arom. H)
[0266] GPC analysis;
[0267] Number average molecular weight (Mn): 19538, weight average
molecular weight (Mw): 20046, polydispersity (Mw/Mn): 1.026
##STR00069##
Example 35
[0268] Using the compound of formula (66) and the compound of
formula (38), a compound of formula (67) was obtained in the same
manner as in Example 13.
[0269] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (24H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.27-3.29 (8H, m, --CH.sub.2--HNCOO--), 3.52-4.23 (3620H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--), 5.19 (4H, brs, --HNCOO--), 5.70 (2.4H, s,
>CH--), 5.82 (1.6H, s, >CH--), 6.95-7.21 (12H, m, arom.
H)
[0270] GPC analysis;
[0271] Number average molecular weight (Mn): 37096, weight average
molecular weight (Mw): 39878, polydispersity (Mw/Mn): 1.075
##STR00070##
Example 36
[0272] Into a 50 mL three-necked flask equipped with a thermometer,
a nitrogen inlet tube and a stirrer were charged the compound of
formula (67) (4.00 g, 0.100 mmol) and dichloromethane (20 g), and
glutaric anhydride (68.5 mg, 0.600 mmol), triethylamine (60.7 mg,
0.600 mmol) and 4-dimethylaminopyridine (3.7 mg, 0.030 mmol) were
added thereto, and the reaction was performed at 25.degree. C. for
6 hours. After filtration, the solvent was distilled off under a
reduced pressure. The residue was dissolved in ethyl acetate (100
g) and crystallized by adding hexane (100 g). After filtration, the
crystals were dried under a reduced pressure to obtain a compound
of formula (68).
[0273] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (24H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 1.97
(8H, quin, --CH.sub.2CH.sub.2CH.sub.2COOH), 2.38-2.46 (16H, m,
--CH.sub.2CH.sub.2CH.sub.2COOH), 3.27-3.29 (8H, m,
--CH.sub.2--HNCOO--), 3.52-4.23 (3620H, m,
--(OCH.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--, --CH.sub.2O--COCH.sub.2--), 5.19 (4H, brs,
--HNCOO--), 5.70 (2.41H, s, >CH--), 5.82 (1.6H, s, >CH--),
6.95-7.21 (12H, m, arom. H)
[0274] GPC analysis;
[0275] Number average molecular weight (Mn): 38021, weight average
molecular weight (Mw): 40873, polydispersity (Mw/Mn): 1.075
##STR00071##
Example 37
[0276] A compound of formula (69) was obtained in the same manner
as in Example 21 using 4-hydroxybenzaldehyde.
[0277] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.32-1.80 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.50-4.24 (5H, m, --OCH.sub.2CH<, --CH.sub.2--OH), 5.71 (0.6H,
s, >CH--), 5.82 (0.4H, s, >CH--), 6.79-6.82 (2H, m, arom. H),
7.31-7.35 (2H, m, arom. H)
##STR00072##
Example 38
[0278] Using the compound of formula (34) and the compound of
formula (69), a compound of formula (70) was obtained in the same
manner as in Example 8.
[0279] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 3.38
(3H, s, CH.sub.3O--), 3.40-4.25 (455H, m,
--(OCH.sub.2CH.sub.2).sub.n, --OCH.sub.2CH<, --CH.sub.2--OH),
5.72 (0.6H, s, >CH--), 5.84 (0.4H, s, >CH--), 6.89-6.91 (2H,
m, arom. H), 7.35-7.39 (2H, m, arom. H)
[0280] GPC analysis;
[0281] Number average molecular weight (Mn): 5142, weight average
molecular weight (Mw): 5255, polydispersity (Mw/Mn): 1.022
##STR00073##
Example 39
[0282] The compound of formula (70) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12 to
obtain a compound of formula (71).
[0283] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (61-1, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.84
(4H, s, -succinimide), 3.38 (3H, s, CH.sub.3O--), 3.40-4.25 (453H,
m, --(OCH.sub.2CH.sub.2).sub.n, --OCH.sub.2CH<), 4.33 (2H, dd,
--H.sub.2O--COO-succinimide), 5.72 (0.6H, s, >CH--), 5.84 (0.4H,
s, >CH--), 6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom.
H)
[0284] GPC analysis;
[0285] Number average molecular weight (Mn): 5257, weight average
molecular weight (Mw): 5373, polydispersity (Mw/Mn): 1.022
##STR00074##
Example 40
[0286] The reaction was performed in the same manner as in Example
8 using the compound of formula (49) and the compound of formula
(69) and then the deprotection of trifluoroacetyl group was
performed in the same manner as in Example 14 to obtain a compound
of formula (72).
[0287] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (6H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.86
(2H, t, --CH.sub.2--NH.sub.2), 3.40-4.25 (453H, m,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--CH.sub.2--OH), 5.72 (0.6H, s, >CH--), 5.84 (0.4H, s,
>CH--), 6.89-6.91 (2H, m, arom. H), 7.35-7.39 (2H, m, arom.
H)
[0288] GPC analysis;
[0289] Number average molecular weight (Mn): 5126, weight average
molecular weight (Mw): 5239, polydispersity (Mw/Mn): 1.022
##STR00075##
Example 41
[0290] Into a 50 mL three-necked flask equipped with a thermometer,
a nitrogen inlet tube, a stirrer and a condenser tube were charged
the compound of formula (71) (2.00 g, 0.400 mmol), the compound of
formula (72) (2.10 g, 0.420 mmol) and toluene (12 g), and the
reaction was performed at 50.degree. C. for 2 hours. Then, Kyoward
200B (0.6 g) was added thereto, and the adsorption treatment was
performed at 50.degree. C. for 2 hours. After filtration,
crystallization was performed by adding hexane (12 g). After
filtration, the crystals were dried under a reduced pressure to
obtain a compound of formula (73).
[0291] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (12H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.27-3.29 (2H, m, --CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--),
3.52-4.25 (908H, m, --(OCH.sub.2CH.sub.2).sub.n--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--CH.sub.2--OH, --HNCOO--CH.sub.2--), 5.72 (1.2H, s, >CH--),
5.84 (0.8H, s, >CH--), 6.89-6.91 (4H, m, arom. H), 7.35-7.39
(4H, m, arom. H), 7.34 (1H, brs, --{right arrow over
(H)}NCOCF.sub.3)
[0292] GPC analysis;
[0293] Number average molecular weight (Mn): 10268, weight average
molecular weight (Mw): 10812, polydispersity (Mw/Mn): 1.053
##STR00076##
Example 42
[0294] The compound of formula (73) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12
and then the resulting compound was allowed to react with the
compound of formula (38) in the same manner as in Example 13 to
obtain a compound of formula (74).
[0295] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (12H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.27-3.29 (4H, m, --CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--),
3.52-4.25 (1356H, m, --(OCH.sub.2CH.sub.2).sub.n--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--), 5.72 (1.2H, s, >CH--), 5.84 (0.8H, s,
>CH--), 6.89-6.91 (4H, m, arom. H), 7.35-7.39 (4H, m, arom. H),
7.34 (2H, brs, --HNCOCF.sub.3)
[0296] GPC analysis;
[0297] Number average molecular weight (Mn): 15296, weight average
molecular weight (Mw): 16856, polydispersity (Mw/Mn): 1.102
##STR00077##
Example 43
[0298] The compound of formula (74) was allowed to react with
N,N'-disuccinimidylcarbonate in the same manner as in Example 12 to
obtain a compound of formula (75).
[0299] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (12H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.84
(4H, s, -succinimide), 3.27-3.29 (4H, m, --CH.sub.2--HNCOO--), 3.38
(3H, s, CH.sub.3O--), 3.52-4.25 (1354H, m,
--(OCH.sub.2CH.sub.2).sub.n--, --(OCH.sub.2CH.sub.2)--OCH.sub.2--,
--OCH.sub.2CH<, --HNCOO--CH.sub.2--), 4.44-4.48 (2H, m,
--CH.sub.2O--COO-succinimide), 5.72 (1.2H, s, >CH--), 5.84
(0.8H, s, >CH--), 6.89-6.91 (4H, m, arom. H), 7.35-7.39 (4H, m,
arom. H), 7.34 (2H, brs, --HNCOCF.sub.3)
[0300] GPC analysis;
[0301] Number average molecular weight (Mn): 15439, weight average
molecular weight (Mw): 17014, polydispersity (Mw/Mn): 1.102
##STR00078##
Example 44
[0302] A compound of formula (54) was subjected to deprotection of
the trifluoroacetyl group in the same manner as in Example 14,
followed by allowing to react with the compound of formula (52) in
the same manner as in Example 41 to obtain a compound of formula
(76).
[0303] .sup.1H-NMR (CDCl.sub.3, internal standard TMS); .delta.
(ppm): 1.40-1.81 (12H, m, >CHCH.sub.2CH.sub.2CH.sub.2--),
3.27-3.29 (4H, m, --CH.sub.2--HNCOO--), 3.38 (3H, s, CH.sub.3O--),
3.52-4.25 (1359H, m,
--(OCH.sub.2H.sub.2CH.sub.2).sub.n--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--), 5.19 (2H, brs, --HNCOO--), 5.70 (1.2H, s,
>CH--), 5.82 (0.8H, s, >CH--), 6.95-7.21 (6H, m, arom. H),
7.34 (1H, brs, --HNCOCF.sub.3)
[0304] GPC analysis;
[0305] Number average molecular weight (Mn): 15279, weight average
molecular weight (Mw): 16822, polydispersity (Mw/Mn): 1.101
##STR00079##
Example 46
[0306] A compound of formula (76) was subjected to deprotection of
the trifluoroacetyl group in the same manner as in Example 14,
followed by allowing to react with N-succinimidyl
3-maleimidopropionate in the same manner as in Example 15 to obtain
a compound of formula (77).
[0307] .sup.1H-NMR (CDCl.sub.3, internal standard TMS): .delta.
(ppm): 1.40-1.81 (12H, m, >CHCH.sub.2CH.sub.2CH.sub.2--), 2.44
(2H, t, --CH.sub.2CH.sub.2-maleimide), 3.27-3.29 (4H, m,
--CH.sub.2--HNCOO--, --CH.sub.2--NHCOCH.sub.2--), 3.38 (3H, s,
CH.sub.3O--), 3.52-4.25 (1359H, m,
--(OCH.sub.2CH.sub.2)--OCH.sub.2--,
--(OCH.sub.2CH.sub.2).sub.m--OCH.sub.2--, --OCH.sub.2CH<,
--HNCOO--CH.sub.2--, --CH.sub.2CH.sub.2-maleimide), 5.19 (2H, brs,
--HNCOO--), 5.70 (1.2H, s, >CH--), 5.82 (0.8H, s, >CH--),
6.15 (1H, brs, --HNCOCH.sub.2--), 6.70 (2H, s, -maleimide),
6.95-7.21 (6H, m, arom. H)
[0308] GPC analysis;
[0309] Number average molecular weight (Mn): 15334, weight average
molecular weight (Mw): 16883, polydispersity (Mw/Mn): 1.101
##STR00080##
Example 47
[0310] Each of the compounds (20 mg) of formula (35), formula (44),
formula (45), formula (47) and formula (48) was dissolved in MES
deuterated water buffer (1 mL) of pD 5.5 and in HEPES deuterated
water buffer (1 mL) of pD 7.4, and allowed to stand in a
thermostatic bath at 37.degree. C. FIG. 1 and FIG. 2 show the
measurement results of hydrolysis rates at pD 5.5 and pD 7.4,
respectively.
[0311] Each of the compounds (200 mg) of formula (41), formula
(54), formula (74) and formula (76) was dissolved in MES deuterated
water buffer (10 mL) of pD 5.5 and in HEPES deuterated water buffer
(10 mL) of pD 7.4, and allowed to stand in a thermostatic bath at
37.degree. C. FIG. 3 and FIG. 4 show the measurement results of
hydrolysis rates at pD 5.5 and pD 7.4, respectively.
[0312] As shown in FIG. 1, the hydrolysis half-lives (t.sub.1/2) of
the compounds of formula (35), formula (44), formula (45), formula
(47) and formula (48) at pD 5.5 and 37.degree. C. were 2 hours, 12
hours, 30 days, 24 hours and 6 months, respectively. Further, as
shown in FIG. 2, at pD 7.4 and 37.degree. C., the hydrolysis
half-lives (t.sub.1/2) of the compounds of formula (35) and formula
(44) were 65 hours and 18 days, respectively, the hydrolysis of
approximately 17% was observed for 18 days for the compound of
formula (47), and no hydrolysis was observed even after 18 days for
the compounds of formula (45) and formula (48).
[0313] As shown in FIG. 3, any of the hydrolysis half-lives
(t.sub.1/2) of the compounds of formula (41) and formula (74) in
which the structure of the cyclic benzylidene acetal linker was
same at pD 5.5 and 37.degree. C. was 2 hours, and was equivalent to
the hydrolysis half-life (t.sub.1/2) of the compound of formula
(35) having the same linker structure. Further, any of the
hydrolysis half-lives (t.sub.1/2) of the compounds of formula (54)
and formula (76) in which the structure of the cyclic benzylidene
acetal linker was same at pD 5.5 and 37.degree. C. was 12 hours,
and was equivalent to the hydrolysis half-life (t.sub.1/2) of the
compound of formula (44) having the same linker structure. As shown
in FIG. 4, at pD 7.4 and 37.degree. C., each of the hydrolysis
half-lives (t.sub.1/2) of the compounds of formula (41) and formula
(74) was 65 hours, and each of the hydrolysis half-lives
(t.sub.1/2) of the compounds of formula (54) and formula (76) was
18 days, and are equivalent to the hydrolysis half-lives
(t.sub.1/2) of the compound of formula (35) and the compound of
formula (44) each having the same linker structure,
respectively.
[0314] From the above, it was shown that when the structure of the
cyclic benzylidene acetal linker was same, the hydrolysis ratio was
same, regardless of the number of polyethylene glycols linked.
[0315] While the invention has been described in detail and with
reference to specific embodiments thereof, it will be apparent to
those skilled in the art that various changes and modifications can
be made therein without departing from the spirit and scope of the
invention.
[0316] This application is based on a Japanese patent application
filed on Mar. 31, 2015 (Japanese Patent Application No.
2015-070659, and the whole contents thereof are incorporated herein
by reference. Also, all the references cited herein are
incorporated as a whole.
* * * * *
References