CA2074509A1 - Polyelectrolyte complex antibacterial agent and antibacterial material - Google Patents

Abstract

An antibacterial agent containing a polyelectrolyte complex prepared by the reaction of a cationic polymer containing N+ atoms in the repeating units with an anionic polymer containing -COO-, -SO3- or -PO3- groups in the repeating units; and an antibacterial material comprising the polyelectrolyte complex carried on a support.

Description

- 1 - 2 ~ 7 ~
DESCRIPTION

?r~ -?~,YT~. CO~ I,E~ ANTIBACTERIAL AGENT
~ D .~NTIBACTERIAL ~ATERIAL

TECHNICAL FT-L~
The present invention relates to an antibacterial agent com?rising a polyelectrolyte complex and an antibacter a~ .,~a-erial carrying the polyelectrolyte complex.

.~CKG~oU~s -?T
Orgar.ir co!~pounds having a positive charge are known to exhibi~ a~ acterial properties, regardless of whether they are low molecular weight compounds or high molecular weisht compv~.nc.s. The application technology utilizing such proper~ies nas spread widely from the medical field to general clothing. For example, quaternary ammonium compounds such as benzalkonium chloride or the like are soluble in water, and so are in themselves used as sterilizing or disinfecting solutions. Conversely, however, since they are soluble in water, the range of usage as antibacterial agents is limited.
Further, anionic groups are introduced to the surface of synthe~ic polymer articles and then treatment with quaternary a~monium bases is performed so as to obtain a material mainsaining an antibacterial property over a sustained long term period. The resulting material is used as a filter material for air filters and dialysis. This technique is also applied to textile materials, and utilized for clothing having antibacterial properties and agents for protecting wound.
As a means of introducing quaternary ammonium to the surface of a polymer material to obtain a material exhibiting sustained antibacterial properties, for example, Japanese Published Unexamined Paten~ Application No. 5g-86584 describes a method for bringing a polymer obtained by ~Q7i~

po'l~eri~ ~ a~ az dl -group-containing monomer into contact ~ a- ~.ueous solution o~ a quaternary ammonium base. F,; ~'ze~ anese Pub'ished Unexamined Patent Ap?licatic. ;:o. -)5-i5~3~12 discloses a method Lor in~roduc'-g -n -~n~sr.ic-croup-containing vinyl monomer to the surfce o- -~ svr1-netlc polymer article by graft pc'~.,eriza~ on ,- ~h- li'.~e, and then treating by a u--~rna~ `^ase q c~ ~ _ . ~ ., ~ . ~ . _ ~ .. .. .
In t^~ ational methods, however, the rate of ir._-oduct ~ '^.e vuat_rnary ammonium base for ma.-._-est'~ r. ibacteriai property was not necessarily su _icien: ~n~ ia_iafa^tory sustenance could not be o~_ained. -~r '^.e-, rne process of production was also cor,plicat_d.
The p~esen.~ inventors engaged in intensive research wi -. the ~b ec~ o obtaining a material having a sustained an~ibacterial a_~ion and having the property of insolubility in solvents (especially water) while maintaining the antibacterial property of a positively charged organic compound, and discovered that the objects can be achieved by a polyelectrolyte complex obtained by reacting a specific anionic polymer and a specific cationic polymer. The present invention is based on the above discovery.

DISCLOSUR- OF T:iE TNVENTION
Therefvre, the present invention relates to an antibacter al agent characterized by containing a polyelectrolyte complex obtained by reacting (A) a cationic polymer containing N+ atoms in repeating units thereof and (~) an anionic polymer containing -COO~ groups, -SO3-groups, or -PO3~ groups in repeating units thereof.
Further, the present invention relates to an an~ibacteria' material characterized by carrying the above-mentioned polyelectrolyte complex on a carrier.
Accorai.ng _o a preferable embodiment of the present invention, as tne cationic polymer (A), use is made of at least one compound selected from the group consisting of:

~ ~ 7 ~ 3 (al) a com~?ound !q~laterna~y ammonium salt polymer) of the general form~lla ( T~:

I R2 Rs _ r ~ R 1 1`~ R4 - _ ~ Xl- ( I ) R3 R6 m wherein R1 ~-.d R1 a~-e, _ndependenr~, an alX~ylene group of 1 to 10 ca-bon a~oms, preferably a straight or branched alkylene gro~p of 2 to 8 carbon atoms, a group of the general forlm~'a:

- R ~

R l 2--:, .
. wherein R1- and R12 are, independently, an alkylene group of 1 or 2 carbon atoms, preferably R11 and R12 being bonded at the p-position, or an arylene group, and R2, R3, Rs, and R6 are, indeDendentlv, an alkyl group of 1 to 3 carbon atoms, or R1 forms, together with the 2 nitrogen atoms and R2, R3, Rs, and R6 in the above formula, a group of the formula:
~

- N N -/
and R4 has ~he same meaning as above, X'~ is a counter ion, and m is a number of 5 or more, ~,7` ~.~
,~
(a2) a com~ou~ d (quate.n.arl a.~monium salt polymer) of the general for..lu'a (Il) -t C H 2 - - C h-t- ~
(11 ) ,~.
wr.erein ~ is a g-cup of tze ~eneral formula:

B - ~ + R 2 2 P~ 2 3 (wherein 3 is a.n alkvlene group of 1 or 2 carbon atoms, 21, R22, and R23 are, inde?endently, a hydrogen atom or an alkyl group of 1 to 3 carbon atoms, and X2- is a counter ion), or A is a group of the general formula:

h~ ' N +--R 2 4 (wherein R24 is an alkyl group of 1 to 3 carbon atoms or a benzyl group and X3- is a counter ion), or A is a group of the general formula:

,' I _ r~ X4 - ~28 ~wherein R2s is an alkyl group of 1 or 2 carbon atoms, R26, R27, and R28 are, independently, a hydrogen atom or an -5- ~r~ "~
al~yl grou~ o~ 1 -o 3 car~on atoms, and X~~ is a counter ion) and n ~s ~ ~ ~. ^r or 1~ or more, (a3) a CO~I~OU~ baslc amino acid polymerj of the ge.neral fo~ 'a ~

r r: H O H H O

- ( i3- C- C ) ( N- C- C ) I ~ I 100-t ( (CH2)V 91 S~ r wherein v is 3 -~ ~, Rgi is a h.ydrogen atom; an a''~yl group of 1 to 4 ca:^Don a~oms; an alkyl grou? of 1 to 4 carbon atoms substituted by a hydroxyl or mercapto grou? or by an alkylthio group of 1 to 3 carbon atoms; or an imidazolylmethyl or indolylmethyl grou?; for exam?le, a methyl, isopropyl, isobutyl, s-butyl, hydroxymethyl, hydroxyethyl, methylthioethyl, mercaptomethyl, 5-imidazolylmethyl or 3-imidazolylmethyl group, R92 is -N+H2Xs~ or -N+HC(NH)NH2X6-, Xs~ and X6- are, independently counter ions, t is 20 to 100, and r is an lnteger of 10 or more, and (a4) a cationic polysaccharide.
~ urther, as the anionic polymer (3), use is made of at least one compound selected from the group consisting of:
(bl) a compound (acidic amino acid polymer) of the general formula (IV):

H H O H H O
I I
- ~ N-- C - C ) -- ( N- C- C ) ( (CH2) U 81 -- COOH q wherein u is 1 or 2, Rg1 is a hydrogen atom, an alkyl group of 1 to 4 carbon atoms; an alkyl group of 1 to 4 carbon -- 2~ s ~
atoms subs-~ a h;~-o~ ' or mercapto group or by an alkylthio 9~ car:sor atoms; or an imidazolyl.~,e~~ - ir.do'~ -eth~l group; for example, a me~hyl, iso -~,, ., s~bu~;l, 5 -butyl, hydroxymethyl, hydroxyeth,l, ~ . e~hyl, mercaptomethyl, 5-imidazolyl~ ` or ~ ~r~ dazo yl~e~..yl grouo, s is 20 to 100 and q is an nteye- o~ 13 or mo-e, (b2) a cc:~.^ound 'acr~lic acld ?olymer) o~ the general formula (v):

?~
t C~ C~ l o O - a~ ( V ) ~ Y CCOR32 J p wherein R31 is a hydrogen atom or a methyl group, R32 is an alkyl group of 6 to 18 carbon atoms, preferably a straight or branched alkyl group of 6 to 14 carbon atoms, Y is a group of a carboxylic acid or a salt thereof, a sulfonic acid or a salt thereof or a phosphoric acid or a salt thereof, or an aryl group containing a group of a carboxylic acid or a salt thereof, a sulfonic acid or a salt thereof or a phosphoric acid o- a salt thereof, a is a numeral of 20 ro 100, preferably 50 to 100, and p is an integer of 10 c- more, and (b3) an anionic polysaccharide.

BEST MODE FOR CARRYING OUT THE INVE`~TION
The polyelectrolyte complex ('r.ereinafter optionally referred to as "PEC") per se which -.ay be used in the present invention is a known substance. As described, for example, in Japanese Published Unex_mined Patent Application No. 49-8581, a polyelec-rolyte complex (PEC) can be immediately formed by mixing a solution of a cationic polymer (a polyelectrolyte having positive charges) a:.. a solut~ of an anionic polymer (a polyelectr~ ..a~ n~gati~e charges). The resulting P~C may be -` s,olvee~. ln a particular three-component solvent (fo:- ec~?'e, ~;ater/acetone/low molecular weight sal. with -- ..art~c~la- c-mrosi~ionj, but is insoluble in a general so ^n~ . ?_C -~ e~.iDi~s a high permeability for variouC oJ."o' c~la- riie~ght com?ounds and so may be used as a d~ is -.e..-^r ane . 3~C may be used to provide various types of materials having various properties according t~v the '.~inG o the starti~.g polymers (polyelectr-~ y~esj, ~he mix ng rat o thereof, the preparatic~. on-~~_cns, o he like. However, it was not kn_rvr. hith-- o mha_ -~' has an ant ~ac~erial property.
In t'~ _se-.~ s~- - c~t or., _n alkyl grou~ of 1 to 3 carbon ato~ ~eans, -o- example, a methyl, ethyl, or n- or i-?ropyl 9~ ?- An a'`.~yl grou? of ~ to 4 carbon atoms means, in ad~ ion tO the above-men~ioned alkyl group of 1 to 3 carbon atoms, for example, an n-, i-, s-, or t-butyl group. An alkylene group of 1 to 2 carbon atoms means, for example, a methylene, ethylene, or ethylldene group. A
straight or branched alkylene group of 1 to 10 carbon atoms means, for example, a methylene, ethylene, propylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, nonamethyle-.e, decamernylene, ethylethylene or ethyltrimethylene group. A straigh_ or branched alkyl group of 6 -o 18 carbon atoms means, for example, a straight alk-~1 group of 6 to 18 carbon atoms, an alkyl group of 6 so 15 carbon atoms substituted with one or more straight or branched alkyl groups c~ 1 to 3 carbon atoms, in particular, an alkyl group of 6 ~o 10 carbon atoms monosubstituted with a straight alkyl group of 1 to 3 carbon atoms. An arylene group mea-.s, for example, a phenylene or naphthalene group. A _arboxylic acid salt group or sulronic acid salt group r.eans, for example, a salt of an alkali metal (for example, sodium or potassium) or an alkal~re earth metal (for ex~ ?le, calcium or magnesium). Further, an aryl group containing a sulfonic .

acid group ~ a s~ -.-reor r~eans, for example, a 2 ~ 3 su' Çonhen ' ~, ., ~
The c ~ esen.c both in the starting cationic po~er (A, ~.~ s ar~ an onlc polymer (~) no longer exis~ in t~ .u^. prepared from the reaction therebet~ic--.. he!c re, the councer ions may be an~y ions so long as -:-ey d~ n~t incerfere with the reaction therebetwee.~. ~-e-2ra~'e csunter ons are halide ions, in particular 2'.'' ~ride, ~romide or iodide ions.
''he ~~ -E-_~ he reac~ion of t:~e cationic polv~er (r. ~:~. . .h~ r c ~ol-y~e~ (B) has a structure that the ~.~ c:-es ~ ne o~tionic ?ol~mer ~re successively bonded to ~.^- -c~d si~_s ('or exam?le, car_oxylic, sulfonic, o:- ,'nosphsr-_ ac~d sites) of the anionic polymer by means of _~u~o~ en-sgy. Namely, the starting polymers are cross~ ed to eacn o~her by ionic bonds and form a gel which - --.so'u~lc in a solvent.
For exa:-,ple, the ?EC produced from the cationic polymer (al) of the general formula (I) and the anionic polymer (bl) of the general formula (IV) has the structure of the general formula (VI):

R2 Rs ~ )1 0 1~ H O
_ N Rl N+ Ri --- ~ ~ N- C - C 1-- ( N- C - C I
l l ¦ I C H ~ I R 8 1 5_¦ ~ V I ) R3 R6 m wherein R1, R2, R3, R4, R5, R6, Rg1, m, u, s, and q have the same meanings as mentioned above, except that 0.25 <
2m/q < 4-0-Further, the PEC produced from the cationic polymer(al) of the general formula (I) and the anionic polymer (b2) of the gcneral formula (V) has the st-ucture of the general formu1a (VII):

-9 2~7 ~ ~3 N'_ Rl ~It~ CI1~ --- C+~CIl~-- ~~ !VI I ) I ' l Y' COO~" 7 R) R~ m ~herein Rl, ~2, ~3, 2~, R5, R6, R31, R32, m, a, and p have the same meanings as mentloned a~ove, except that 0.25 <
2m/p < 4.0, an- Y is a -C^O~, -S03- or -P03~ group, or Y' is an aryl g-oup containing a -COO~, -S03- or -P03 group.
Further, ~he P~C produced rrom the cationic polymer (a2) of the general formula (II) and the anionic polymer ~bl) of the general formula (IV) has the structure of the general formula (VIIIj or (IX):

H H 0~ H H C~
t C H 2 C H ~ n --- ( N- C-C I-- I N- C-C I _ I (C~ S ~ 00-5~ (Vlll~

--( C H 2 C H ~ r~ _ - ( N-- C--C )-- I N- C- C I
5 I 100-~ (IX) - coc~~ 8 1 q N+~
R2~

1,7 - 'O-In the ar~ or~iulae (VlII) and ~IX), R21, R22~ R23, R2~, R8l~ n, u, s, and q ha-~e the same meanings as mentioned a~o-,e, e~ce?t that 0.25 < n/q < 4Ø
The PEC ?roduced from the cationic polymer (a2) of the general forr.ula !TT) and che anionic polymer (b2) of the general formula (~J has che structure of the general formula (X) or (XI):

--t C H ~ -- C H 7 n ~ C '~ --C7 ~ Crl~-- C7~ ( X ) ~1 ~ Y COORJ~ J, --( C H 2 C H ~ n _ -t CH~ --f, t ~-- I " o o ~ ( X I ) ~ Y' COOF~I, ~

~ NJ
,' I
R2~

In the above formulae (X) and (XI), R21, R22, R23, R24, R31, R32, Y', n, a, and p have the same meanings as mentioned above, except that 0.25 < n/p < 4Ø
The PEC produced from the cationic polymer (a3) of the general formula (III) and the anionic polymer (bl) of the general formula (IV) has the structure of the general formula (XII):

2 r~7 ~

H ~f O1~ ~i _ ~ C - C I ~ ~1- C - C i ~ - I ~1- C - C I-- ~ ~1- C - C ) _ ~ c ~ ~ ~ vI 1 0 0 ~ I ~ C ~ U I I O û~ ~ X I I ) R ~ _ C O O - _ q wherein Rg1, RP~ r, ~ v q, u, and s have the sarne meanings as me~ cned a~ove, excep~ that 0.25 < r/q < 4.0, and Rg3 is a r~!~:~2 or -`~1-t.~.C(NH)~lH2 group.
The PEC pr^duce_ f~om the cationic polymer (a3) of the general formula ( T-- ) ar. ~he anionic polymer (b2) of the general formula ~,~) 'r.as ~'ne struc~ure of the general formula (X-~

C-C~ C-C~ -C~ (Xlll) ~CH,I V R9 ~ y - R"

wherein Rgl, R93, R31, R32, Y~, r, t, v, a, and p have the same meanings as mentioned above, except that 0.25 < r/p <
4Ø
The PEC which can be used as an antibacterial agent in the present invention is not limited so long as being a solid insoluble in a solvent. The average molecular weight thereof is no~ particularly limited, but the number of ion sites in the PEC is generally 10 to 1000, preferably 20 to 100. The average molecular weights of the starting cationic polymer (al) to (a4) and the starting cationic polymer (bl) to (b3) are not particularly limited, but as to the preferable range thereof, _ in the general formula (I) [the cationic polymer (al)] is 5 to 500 (in particular 5 to 100), n in ~he general formula (T ) [the cationic polymer (a2)] is 10 to 1000 (in par~icular 10 to 500), and -12~ u--r in the ger.eral ormuld (III) ~the cationic polymer (a3)]
is 10 to lCOC ! s. par~iic~ r 1(, to ~30). Eurthe , g in the general fo~r~ `r~-. ca~ios.ic polymer (bl)] is 10 to 1000 (in pa-clc~ 10 tO 500,, and D in the general formula (vj ,~'se _a~ion:c pol;r.er (~2)~ is 10 to 1000 (in particular '~ tO _~'i) .
As examoles of the cationic poiymer (al) of the general forrn la (_,, there ma- ~e mentioned ~uaternary polyethyler.eirrine chlo{lde, poly(N,~ Jr',N'-tetramethyl-alkylene-p~ lerae dia~ol~ium dichlo-ide), poly(N,~J,N',M'-tetramethyl-alkylene-dialrrmoni~rr dichloride), poly(N,N-dime~hyl-3-..-;_ro.;-ipropylarnmonium ch oride), polv(2-hydroxy-3-methacroylo.Yypropylr~imethyla.rrmor.i~l chloride!, pol~(2-methacroyl~-e~hy'tr-i.rretn.ylammo~i~rn chloride), poly(glycicyitrimechyl-ammonium chloride), poly[(dimethyliminio)ethylene(dimethyliminio)-me~hylene-1,4-phenylene~,e~hylene dicnloride] iin general known as 2X], poly[(dimethyliminio)hexamethylene(dimethyliminio)-methylene-1,4-phenylenemethylene dichloride] [in general known as 6X], poly[(dimethyliminio)hexamethylene chloride]
[in general known as 6,6], poly(N-ethyl-4-vinylpyridinium bromide), poly(dimethyldiallylammonium chloride), or the like.
As examples of the cationic polymer (a2) of the general formula (II), there may be mentioned poly(vinylbenzyltrimethylarnmonium chloride), polyvinylpyridinium chloride, poly(N-benzyl-4-vinylpyridinium chloride), or the l-ke.
As examples of the cationic polymer (a3) of the general formula (III), there may be mentioned polylysine, polyarginine or copolymers thereof, or copolymers of the monomers of said polymers with glycine, alanine, phenyl alanine, tyrosine, valine, leucine, isoleucine, serine, threonine, methionine, cysteine, histidine, proline, and/or tryptophan, or the like.
Examples of the cationic polysa^charide (a4) are as follows:
(1) Chi~osan and derivatives hereof:

, 2 ~ J . ~, ~

2-~ ~

' ~ 2 ~ 6 6 q2 6 wherein R66 -s a `n.ar^^en a~o.m or an acetyi group, X7- is a councer ior;, ~`e _eace~,lizacion deyree is 50 to 100 pe-cent, pre-^~a~~y ,0 ~^ 10^~ percen-, q26 is 20 ~o 3000, preferably -0 ~.30. Ic is ..o~ed ~hat in the a~ove formula, the ~' a-cms r the -~+H2R6o groups and the anionic groups -~. the ar.ioni- ?ol~er are bonded to each other.
(2) Dlethvl~milloethyl Derivative of Neutral Polysaccharide:
As a neutral polysaccharide, there may be mentioned dextran, cellulose, mannan, starch, agarose or the like.
The degree of diethylaminoethyl substitution of the above derivatives is O.S to 2.0, preferably 0.7 to 1.5 groups per one sugar residue. The degree of polymerization is 50 to 5000, preferably îOO to 1000. It is noted that the nitrogen atoms in the diethylaminoethyl groups and the anionic groups in the anionic polymer are bonded to each other.
Examples of ~he anionic polymer (bl) of the general formula (IV) are polyglutamic acid, polyaspartic acid, or copolymers thereof, and copolymers of the monomers of said polymers with glycine, alanine, phenylalanine, tyrosine, valine, leucine, isoleucine, serine, threonine, methionine, cysteine, histidine, proline, and/or tryptophan, or the like.
It is no~ed that the polyamino acids of the general formulae (III) ana (IV) may be prepa-ed by the general acid . . .

2'~7 ~

anh~dride monomer mechod, the active esterification method, the Merryfield .,~ecr.cd, oc the like.
As examples of the anionic polymer (b2) of the general formula (V), there ma~ be mentioned polyacrylic acid, polym.ethacrylic acid polyitaconic acid monoesters, poly.-,.aleic acic r._noescers, polyv~nylsulfonic acid, polystyrene sul o-.ic acid, ar.d coooly~ers of two or more monomers const u ir.~ e above polymers, and copolymers of such monomers with carboxylic acid derivatives having C6 to C1g alkyl groups bonded by esterification to the carboxylic grou?s of the ab^.e ~.OQ.Ome.S.
Exam?les o~ he arion c ?olysaccharide (b3) are as follo~.;s:
al~rcr. c .~.cic. ar.d Der vatives Thereof ., , ~ \~ .
OH NHCOCH3 ql wherein R41 is a hydrogen atom or an alkaline metal (for ' example, sodium or potassium), but disappears in at least a portion of the re?eating unics in che PEC prepared by the ~ reaction with a cacionic polymer, and q1 is 100 to 12000, .. preferably 200 to 8000.
(2) Algir.ic .'.cid and Deriva-ives Thereof i ~R~2 ` to ~ ~o~
COoR42 q2 --:, .... .. .. . . .. .
'',"`:: , ' , ' ' . ' - ~ ' -' , . ~
, : .

- 5- 2 ~ 7 !1 `", ~

~herein R~2 is a hJrl-oye?. atc.n or an alkaline metal (for e~æmple, sodi~r.i~-~r ~o~assium), DUt disappears in at least a portion of the -eDea~ing units in the PEC prepared by the reactio.n 1~ith t:-.e cationic pol~.ner, and q2 is 100 to 10000, prer era'~ly 2 J ~ r C C .
!3~ Cso-.~ . Sl flr-- ~.cid A

H ~ ~

O.i NHCOCH q3 wherein q3 is 10 to 100, preferably 10 to 50, and at least a portion of the COOH and/or S03H groups in the formula is converted to COO~ and/or S03- groups by reaction with the cationic polymer.

(4) Chondroitin Sulfuric Acid C

~ ~~

OH NHCOCH3 q4 wherein q4 is 10 to 100, preferably 10 to 50, and at least a portion of the COOH and/or S03H groups in the formula is converted to COO~ and/or S03- groups by reaction with the cationic polymer.

(5) Chondroi~in Sulfuric Acid 3 (Dermatan Sulfuric Acid) and Derivatives Thereof ,, .

-i6- 2n7~

~~, ~"~'1``

` NHCOCH3 NHCOCH3 q5 wherein qs is 20 to 100, preferably 40 to 50, and at least . a portion of the COOH and/or S03H groups in the formula is , converted to COO- and/or S03- groups by reaction with the ~, cationic polymer.
~ (6) Chond~o tin Sulfuric Acid D and Derivatives ;~ Thereof ... .

~ COOH CH20SO3H COOH CH20SO3H
~ 0~o~\0 ~NHCOCH3 OSO3H NHCOCH3 .
wherein q6 is 10 to 500, preferably 20 to 100, and at least ,a portion of the COOH and/or S03H groups in the formula is converted to COO~ and/or S03- groups by reaction with the cationic polymer.
(7) Chondroitin Sulfuric Acid E and Derivatives .
Thereof - .
, ,~ L_ _~
:~ ~ o\ ~ V\ / j , HO3S/ O
` ~ ~ o ~ o~

~ NHCOCH3 NHCOCH

' .,,,. V.'.. . -."~ ' '~, . : '' ' ', ' ~'-..

1 ~ 2 ~ 7 ,i,~", ,, ~ ,,J
wherein q7 is 10 to 300, preferably 20 to 100, and at least a portion of ~he COOH ar.d/or SO3H groups in the formula is converted to CCO~ and/or SO3 groups by reaction with the cationic ~o'y-ier.
(8) r.epa~an Sul-uric .icid and Derivatives Thereof COOH CH ~OH CH20SO,H
j K~ r~oO~ r~

wherein q8 is 7 ~o 200, preferably 10 ~o 100, and at least a portion of th.e CCOH and/or S03H groups in the formula is converted to COO~ and/or SO3- grou?s bv reaction with the cationic polymer.
(9) Heparin and Derivatives Thereof ~ ) O ~ Lo~ /1 ot . NHCOCH3 OSO3H NHCOCH3 wherein qg is 100 to 500, prererabl~y 100 to 300, and at least a portion of the COOH and/or SO3:i groups in the formula is converted to COO~ ar.d/or SO3- groups by reaction with the cationic polymer.
(10) ~-Carragheenan and Derivatives Thereof ~ . .

- 13 - 2 ~ J

C'~20~1 CH2 ~5~ ! o~ G~I

OH. CRso qlO
wherein Rso is a hydrogen atom or S03H group, qlo is 100 to 10000, preferab'y 100 to 500, and ar least a portion of the S03H groups in the ormula is conve-ted to S03- groups by the reaction w~th ~he ca~ onic pol~-.er (11) ~-Carragheenan ard ~eriva-ives Thereof CH20H CH20Rs Lo~ ~~ Lo ORsl OH qll wherein Rsl is a hydrogen atom or S03H group, qll is 100 to 10000, preferably 100 to 500, and a~ least a portion of the S03H groups in the formula is conve~ted to S03- groups by the reaction with the cationic polymer.
Further, after carboxymethylation, sulfation, phosphoriration or the like, neutral natural polysaccharide may be converted to and used as the anionic polysaccharide (b3). Examples of such modified po`ysaccharides are as follows:
(12) Cellulose Deriva~ives - 5- ~2 ~

C~JzO~tsz ~o~

~ts2 J
~ C~12 ;`
wherein Rs2 is a hyd~osen a-om, o- a carbox-rmethyl, sulfuric acid o- _hossh ric acid g-oup, ql2 is '00 to 15000, preferably 200 ~o 5000, ar.~ at least a portion of the COO~, SO3H and/or ~ rou?s n the formula is converted to CCO , SO3 and/or rO3 groups by the reaction with the cation.ic ?o~ymQr.
(13) Cr.i~ir. 3eriva~ives {'~/
NHCOCH

~' wherein Rs3 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, ql3 is 50 to 8000, preferably 100 to 5000, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to COO , SO3- and/or PO3 groups by t:~e reaction with the cationic polymer.
- (14) Carboxymethylstarch and 3erivatives Thereof - ~

-20- 2~ 7~

ORs4 - -~\
,s ~
.,, . ORsq ,,` ~ qlq wherein Rs4 is a hydrogen atom, or a carboxymethyl, - sulfuric acid or phosphoric acid group, q14 is 100 to 8000, ~' preferably 200 ~o 5000, and at least a portion of the COOH, -~ SO3H and/or PO3H grou~s in the formula is converted to COO-, SO3- and/or PO3- groups by the reaction with the cationic polymer.
(15) Amylose Derivatives ~ORs s ~ -o~ "
~ Rss ''~''' q~S
, .
~j wherein RS5 is a hydrogen atom, or a carboxymethyl, .~ sulfuric acid or phosphoric acid group, q1s is 100 to 8000, preferably 100 to 5000, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to ~; COO-, SO3- and/or PO3~ groups by the reaction with the ;l cationic polymer.
` (16) Amylopectin Derivatives :~

, :~,'' .
.
.

" .
,' ~, ,; . .~ :-: ~ -. -- ~ : - ~ :
,, : .
,' - ~ . ~, ~ . ' -- : . .

, . .

-21- 2~7~f~
/ c~20RS6 Iko~l ----`~r ' l I H20Rs6 CH2 CH20R56 ~ 1~

ORs6 ~56 R56 ql6 wherein Rs6 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q16 is 100 to 100000, preferably 100 to 10000, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to COO , SO3- and/or PO3 groups by the reaction with the cationic polymer.
(17) ~-1,3'-Glucan Derivatives (For Example, Cardran) / q l 7 wherein Rs7 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q17 is 50 to 1000, preferably 100 to 300, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to COO~, SO3- and/or PO3~ groups by the reaction with the cationic polymer.
(18) ~-1,2'-Glucan Derivatives , :;. - . . . . . . . . . . .
", .. . - , . , : . : :

", "
: : : . : ~ ~:

-22- 2~7/~

CH2ORs8l ~\~
~\ OR58 Rss \~
I
qla wherein Rs8 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q18 is 100 to 4000, preferably 100 to 3500, and at least a portion of the COOH, S03H and/or P03H groups in the formula is converted to COO , S03- and/or P03~ groups by the reaction with the cationic polymer.
(19) ~-1,3'-; ~-1,6~-Glucan (For Example Lentinan, Schizophilan, Coriolan) Derivatives ~OR s 9 ~ O
~ ~1 :~

~ 0 ~l--o~ 0~
,'Rs90 RsgO RssO .' ORsg ORsg ORsg ql9 , wherein Rsg is a hydrogen atom, or a carboxymethyl, ,~ sulfuric acid or phosphoric acid group, q19 is 100 to ; 100000, preferably 100 to 50000, and at least a portion of ~ the COOH, S03H and/or P03H groups in the formula is . .
,.

.
.

`~ '-'' , ,, . ~ :
,,,: . , , ~ ~ ' :

23- 2r!~ 3 .` .
converted to COO~, SO3- and/or PO3- groups by the reaction ;~ with the cationic polymer.
(20) Dextran Derivatives wherein R~o iS a hydrogen atom, or a carboxymethyl, . sulfuric acid or phosphoric acid group, q20 is 100 to ~ 300000, preferably 200 to 100000, and at least a portion of the COOH, S03H and/or PO3H groups in the formula is i~ converted to COO , SO3 and/or PO3 groups by the reaction with the cationic polymer.

:~

. .

.~

., , .

, ., ., , ;~
~' ~

. ;, . .,.:, , . . .
.. . . . .

(21) Pullulan ~erivati~ves `~ ~
o~ I ~oT
~ I ~ I
., ~ o,~
~o ~ o~
o ~
.~_ o ~s I -~

'~ 7 ~o ~o ~1 ~
~o .~

,................................ .

:

-2~- ~ r~ 7 ,1 . ".j wherein R61 is a h~droger, atom, or a carboxymethyl, sulfuric acid or p,~os?horlc acid group, q21 is 300 to 2000, preferably 500 to ,~9, ar.~ at least a portion of the COOH, SO3H and/or PO3H groups in ~he formula is converted to COO~, SO3- and/or ~o3- grou?s by the reaction with the cationic poly~er.
(22) Agarose ~eriva~ives ~ o \ O

?`62 q22 wherein R62 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q22 is 20 to 200, : preferably 20 to 100, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to COO~, SO3- and/or PO3~ groups by the reaction with the cationic polymer.
- (23) ~-1,4'-Galactan Derivatives '~ _ -0~ ~ _ OR63 q23 wherein R63 is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q23 is 50 to 200, preferably 50 to 200, and at least a portion of the COOH, SO3H and/or PO3H groups in the formula is converted to COO~, SO3- and/or PO3~ groups by the reaction with the cationic polymer.

-26- 2 ~
(24) Manna~ De~iva~i,es C H 20 R ~
~o`~

a 2 ~

wherein R64 is a hydrogen a~cm, or a carboxymethyl, sulfuric acid or phospnor c acid g-oup, q24 is 50 to 5000, preferably 100 to 3000, a-.- a~ leas a portion of the COOH, S03H and/or ?03~ ~roups in -.~e or ula is converted to COO~, S03 and/or ?03 grc ?s by the reaction with the cationic polymer.
(25) Inulin Derivatives I~
R6 5OH2 1C/ \ O
~\ OR 6~
~/ CH2 I

OR 6 s ~
q2s wherein R6s is a hydrogen atom, or a carboxymethyl, sulfuric acid or phosphoric acid group, q2s is 20 to 100, preferably 20 to 80, and at least a portion of the COOH, S03H and/or P03H groups in the formula is converted to COO~, S03 and/or P03~ groups by the reaction with the cationic polymer.
The polyelectrolyte complex (PEC) used in the present invention may be prepared by a usual method. More particularly, the reaction of aqueous solutions of the above-mentioned cationic polymers and anionic polymers (10-5 mole/liter to 10-2 mole/liter) is carried out in aqueous solution within 0.25 to 4.0, preferably 0.4 to 2.5, .

, -27- 2 ~ 7 ~ J 3 of a concentration ratio of the cationic sites of the cationic polymer and the anionic sites of the anionic polymer (cationic sites/anionic sites). If the concentration ratio of the cationic sites and anionic sites (cationic sites/anlonic sites) goes out of the above range of 0.25 to 4.0, it becomes àifficult to form the polyelectrolyte com~lex (?~C). The above reaction has a relatively high reactl~it~. ~herefore, the pH of the solution, the ionic strength, the temperature or the like may vary in a relati~iely wide range, but in general the reaction is car-ie~ out at a pH of 3 to 9, an ionic strength of 0 to 1.0, and a temperature of 20 to 40C.
The charge balance of the PEC used in the present invention is -6 to +3, pre erably -4.5 to +4.5. In the present specification, the term "charge balance" means the charge state of the PEC, expressed by the concentration ratio of the cationic sites of the starting cationic polymer and the anionic sites of the starting anionic polymer. For example, when the concentrations of the cationic sites of the cationic polymer used and the anionic sites of the anionic polymer used are equal to each other, the charge balance of the PEC produced becomes +0. If the concentration ratio is greater than that of the above case (namely, the concentration of the cationic sites is higher), the charge balance becomes positive, while if the concentration ratio is smaller (namely, the concentration of the anionic sites is higher), the charge balance becomes negative. Further, when the concentration ratio is 1.5, the charge balance becomes +2, while when the concentration ratio is 0.5, the charge balance becomes -3.3. The charge balance may be easily adjusted by changing the amount used of the aqueous solution of the cationic polymer and the aqueous solution of the anionic polymer, each having the equal concentration, respectively. By adjusting the charge balance, it is possible to obtain an excess cationic or anionic state.
The polyelectrolyte complex (PEC) is obtained as a gelanious precipitate from the reaction solution.

- . . - , - . : . : , - -, . ,: ' ~ ' .. . ~ , ~, , . . . , -- .
.

2 ~ 7 ~ ~tJ ~
-2a -Therefore, tne P~C Inay be used in the form of the resulting gelanious precipltate, or directiy shaped and processed to a suitable ~orm (or e~ample, a ~iber, film, sheet, block, latex, or gel) and ~s.ed as an antibacterial material in a wet or dry s~a e. ~~lrther, the ~EC can be deposited or adhered on almcs~ al~_ materlals, and thus can be cGated on a suitable carrier tO prepare an antibacterial material.
If an antibacte.ia' proper~y is im?arted to a liquid per se depending on the a~v?licacion thereo~, the PEC reaction solution may also be àirectly used in the form of suspended liquid.
As the carrie- made of an orsanic material, there may be mentioned, for exaln?le, an organic polymer material, such as, synthetic or na~ural resin, synthetic or natural rubber, synthetic or natural fibers, bi.opolymer materials, leather, ~ood, ?ulp, and paper.
As the syn~he~ic resin, there may be mentioned, for example, hydrocarbon polymers (for example, polyolefin, polyethylene, polypropylene, polybutene-1, poly-4-methylpentene-1, polystyrene, polyacetylene); halogenated hydrocarbon polymers (for example, polyvinyl chloride, polyvinylidene chloride, fluorine resin); unsaturated alcohol or ether polymers (for example, polyvinyl alcohol, poly~inyl ether, polyphenylene oxide, polyphenylene sulfide, polyacecal, polyether, polvvinylbutyral, polyether sulfonepoxy resin); unsaturated aldehyde or ketone polymers (for example, phenol and urea resin); unsaturated carboxylic acid polymers (for example, acrylic resin);
unsaturated ester polymers (for example, polyvinyl ester, polyacrylate, wholly aromatic polyester, polyethylene terephthalate, polycarbonate, poiybutylenediallylphthlate, unsaturated polyester resin); unsaturated nitrile polymers (for example, polyacrylonitrile, ABS resin, AAS resin, AES
resin); unsaturated amine polymers (for example, polyvinylamine, polyimide, polyamide, melamine resin, polyethyleneimine, polyurethane); and also silicone resins, the copolymers or blended resins o- above-mentioned ?, ~7 ~

polymers, and the.mGplastic elastomers. Example of the natural resins is cellulose derivative resins.
As the synthecic rubber, there may be mentioned, for example, s~yrene-r~tadiene, butadiene, isoprene, nitro, chloroprene, butyl, erhylen.e-prop~lene, acrylic, chlorinated pol~e~~.y'ene, 'uoro, silicone, urethane and polysulfide rubber.
As the synthe~ic fiber, there may be mentioned, for example, regenerated cellulose acetate (viscose rayon, cuprammonium rayo.-`, -riacetate, polyamide, acryl, vinylon, vinylidene, pol~r~/ n~,-~ chl ride, polyester, polyethylene, polypropy'ene, ~o yb2nzoate, pol~cral, aramide phenolic fiber, polyurethar.e flb2r, fluorine fiber, polyvinyl alcohol, carbon f _er, and silicon carbide fiber. Examples of the na~ural fiber are cotton, silk, wool, hemp, and wood.
As the carrie-, tnere may also be used inorganic materials, for example, glass, minerals (for example, asbestos), enamel, cement, ceramics, artificial stone, and metals (for example, iron, steel, non-ferrous metals, alloys). The shape and form of the carrier are not particularly limited and may be any of a fiber, filament, film, sheet, woven fabric, nonwoven fabric, bar, string, sphere, powder, granule, porous body, hollow body, aggregate, foam, gel, and the like.
The PEC can be carried on the carrier by any known method, for example, coating, spraying, dipping, or the like. It is sufficient to merel-y bring the PEC solution into contact with the carrier. For example, the PEC can be strongly adhered on the bottom of a container by mixing the aqueous solution of the cationic polymer and the aqueous solution of the anionic polymer in a reaction vessel, then transferring the reaction solution immediately to said container, allowing to stand overnight or so to sufficient precipitate the PEC, then removing the supernatant from the container, washing with physiological saline solution and distilled water about one to three ~imes, and drying at 60 to 100C for 6 to 12 hours for annealing. The entire , ' ', -; - , : .
.

30 2 ~ r! ~r;

inside surface of a container can be strongly coated with the PEC, by agitating the solution while rotating overnight or so to sufficiently precipitate the PEC, then removing the supernatant from the container, washing and annealing in the same manner as above.
Further, in the case of a fiber, bead, woven fabric or the like, such materials may be treated in the same manner as mentioned above, after dipping in the PEC solution overnight or so. Further, as described in Japanese Published Unexamined Patent Application No. 50-63096, it is possible to prepa.e the PEC in the presence of a water-soluble organic solvent (for example, a mixture of water, acetone, and sodium bromide), and directly use the reaction solution as a coating agent for coating, spraying or dipping.
hen the PEC is carried on a highly hydrophobic surface of the carrier (for example, a polycarbonate carrier), it is preferable to perform the treatment for imparting hydrophilic property to the surface (for example, treatment with hypochloric acid, organic solvent, plasma, or ultraviolet radiation) in advance.
The resulting carrier carrying the PEC thereon may be directly used as an antibacterial material without further processing. Further, such an antibacterial material may be used to prepare various antibacterial products. As examples of the antibacterial materials which can be used without further processing, there may be mentioned a textile material (for example, fiber, filament, woven fabric, or nonwoven fabric) carrying the PEC on at least a part of the surface thereof (preferably, the entire surface thereof), for example, PEC carrying gauze, absorbent cotton, or fabric (for sterile clothing products for medical, sanitation, or beauty use~. From the above PEC
carrying fiber materials, it is possible to simply prepare, for example, masks, eye bandages, bandages~ sheets, absorbent pads (for example, for the ears, nose or mouth, or menstrual tampons), and napkins.

, ,,, -., ~
:, .

, - ' .:

, -31- 2~7~
Further, it is possible to prepare various types of sterile clothing, for example, underwear (undershirts, undergarments, socks, etc.), baby linen products (for example, baby underpants, bibs, swaddling clothes, singlets, etc.), handkerchiefs, corsets, girdles, brassieres, swl~mins suits, surgical operation garments, surgical and patient use aprons, life-saving devices, diving suits, laboratory clothing, protective clothing (surgical gloves), masks, and surgical hats.
The antibacterial products pre,ared using the PEC
carrying antibacterial materials are not particularly limited, so long as -he suppression of.the proliferation of microorganisms is desired therein. ror example, there may be mentioned medical devices, sanitation devices (for example, hospital-use bed covers, sheets, sterile clothing, bandages, diapers, eye bandage gauze, tampons, contact lenses, contact lens containers, pharmaceutical storing containers, blood transfusion containers), food devices (for example, food packaging materials and food storage containers), household devices (for example, devices for dining table use, kitchenware such as a bottom sheet for a cupboard, sanitaryware such as toiletseat covers), barber and beauty shop devices, devices where slime easily occurs (for example, dialysis membranes and filter materials), and physicochemical machinery and equipment (for example, humidifiers, washers, and constant temperature tanks), etc.
Particularly preferable examples of the medical devices (more preferably disposable medical devices) as the antibacterial products prepared from the PEC carrying antibacterial material of the present invention will be mentioned hereinafter. The preferred carrier materials therefor are shown in parentheses.
As general medical and nursing devices, there may be mentioned, for example, adapters [or connectors]
(polyethylene, polypropylene, or polyamide), irrigators (polyvinyl chloride), indicators (Japanese and western style paper), aprons (nonwoven fab--^s/, diapers (polypropylene fibers, nonwoven fab-ics, paper, cotton, ,- ~ . ~ . . .. .
- . .

.
:- , , -,.......... .. .
, ~ .

~ 5 -~

polyamide, pul? , gd-lze ~r!or;oi~r ~abrics, paper, paper cotton, pol~ia.~s~- ac-~ ;, po'~e--,ter), cups [specimen con~ai-.erj] ~ o~ e, rjo'' er'-..lene, polystyrene, paperJ, cat',ete-a '~~ ,li ir. ' chloride, rubber, silicone, ?~ prc-,~ler.e, polyamide), covers (nonwoven fabr~_-j, p_'ie-ai'ene), __-fs (polyvinyl chloride, ru'ober~, e e ba.r.dages (cauze, nonwoven fabrics, synthet c fi e~s , e~.e-,la -e ~-es !s,rthetic fibers), caps (nonwoven fabrics, pa?er) suctior. devices (polyvinyl chloride, plas[lcs r~bbe-~, clamrs ~clips] (sponge, ru'^be- ~t,~'s -~ ~ chlorid~ acetal , , , . . _ ~ _ , ., _ ~ . . , ~ _ . ~ . . _ . . _ _, resins), ex,-.min; i~-l c' G_ `i`-`9 ~ o.~w Jen fabrics), coils [for hearing b~cs_, ipoli-,in~l _hlcride, polypropylene), oxygen tents (po';~e~'r;-~er.e, pol;~ r.yl chloride), three-way plugs (polyamide, pol~ace-al, Derl n, polyvinyl chloride, polymethyl penter.e), ~-t f'clal ncses (paper, polypropylene), s~oppers ~polyeth~:ene, polystyrene, polypropylene), blood transfusion sets (polyethylene, polystyrene, polypropylene, rubber, polyvinyl chloride, metals), towels (nonwoven fabrics), cavity scopes (polyvinyl chloride), syringes (rubber, polypropylene, medical use silicone oil, polymethylpentene), needles (polyethylene, stainless steel, polypropylene, polyvinyl chloride), hearing aids (polyvinyl chloride), proctoscopes (polyvinyl chloric,e), tapes [stick' ,r,g plasters] (acrylics, polyester, polyethylene, cotton, Japanese paper, polyvinyl chloride, polyamide, rayon), T-bancages (nonwoven fabrics, paper), gloves (polyethvlene, pol~ inyl chloride, rubber), instillators (polyethylene, polypr_-ylene), trays (compressed pulp, paper), urinals ~polyester, polyethylene, polypropylene, polyvinyl chloride, .`BS, rubber), name bands (polyvinyl chloride, polyethylene), pus basins (paper, pulp), bags (polyvinyl chloride, s ainless steel, polyethylene, polystyrene, rubber, ?aper), pads [cotton]
(cotton, gauze, polyester beads, r. nwoven fabrics, paper), acupuncture needles (s~ainless stee:), splints (polyisoprene), belts (spandex), c_st bandages (cotton, gauze, knitted fabrics, nonwoven f_brics, plaster, -3 ~
polyamide), mouthpleces (pGl~lstyrene, paper), masks (poly?ropylene, ~o~~et;lyiene, pol~-arnide, polyvinyl chloride, no~..^wo~,es. fa~rics), !rats loolyethylene, aluminum, adhesives), ~nc.-e ers ~ool~s ;re.ne), cotton balls (cotton), cot_on s~;aDs (~,ir.i~e '~i~ch rnaterial), finger sacks (polyethyiene, ~ er , ~ /el -n~; ne~dles (stainless steel, polyvinyl cnloride, ABS, rubber, metals, polyethylene, pcl-~,~ropylene, fluororesins), connecting tubes (polyvinyl chloride, ?olyethy]ene, rubber, metals, polypropylene, polyamide), ar.d che like.
~ urther, ~s ~nesth.eti^ ard s~rgical devices, there rnay be mensioned, fc~- ei:,r.:rle, int~a~uâers for use of vascular injection (poly~ . C`'llCr' de, ~ , polyethylene, polypropylene, ru~be-, metals, Tef~lon), airways (polyvinyl chloride, ethylene/vinyl acesate copolymer), blepharostats (tantalum), go~r.s (nonwoven fabrics), catheters (polyvinyl chloride, polyvi-.yl chloride with mixed silicone, latex, stainless steel, Teflon), shoe covers (nonwoven fabrics), cuffs (latex), caps (nonwoven fabrics, cellulose), suction devices [suction tubes] (polyvinyl chloride, polyamide, polypropylene), pharyngoscopes (polyvinyl chloride), connectors (polyethylene), vascular injection sets (polyethylene, polypropylene, polyacetal, Teflon, polyvinyl chlorlde, metals, silicone), towels (nonwoven fabrics), counter electrode ?lates (aluminum foil, copper, stainless steel foil, Bose paper, stainless steel plate), tapes (nonwoven fabrics, filaments), gloves (rubber, polyethylene), dra?es (polyvinyl chloride, polyethylene film, nonwoven fabrics), drains (polyvinyl chloride, rubber, silicone rubber), biopsy needles (stainless steel, PBS, polyvinyl chloride, polystyrene, metals), suture thread (silk, polyamide, polypropylene, polyester, stainless steel, catgut), masks (polyester, nonwoven fabrics, glass fibers, polystyrene), scalpels (stainless steel, polyvinyl chloride, ABS), and the like.
Furthermore, as examination and examination room devices, there may be mentioned, for example, cover glass (glass), blood sample tubes (glass, acrylics, polypropylene, natural ru`~be-, synthetic rubber), blood sample bottles (oo y?ropylene, riolyethylene, polystyrene), test tubes (pol~prop~lene, polyethylene, styrene resins, glass), petri dishes !polyst~rene, paper, glass), Spitz tubes (polyprop~lene, pol~st~rer.e, acetyl cellulose, acrylicsj, plungers (polyecri enei, slide glass ~glass), tapes (paper), elecsrodes [for electrocardiograph etc.]
(synthetic fiber, ?aper, lea ~Ji-e, polyethylene, gel), incubators (polyethylene, glass, acrylics, synthetic rubber, polystyrene), beakers (polypropylene), pipettes (glass, polypropylene), labels (paper), and the like.
Further, as ar ificial organs and artificial kidney room devices, the-e ~.a~ be mentioned, for example, catheters [cannulas, !~eflon si'icone rubber, polyvinyl chloride, polyetr.~ene, DO' ~ r-~ylene, cotton), blood circuits (rubber, polyvinyl chloride, polypropylene, polyamide, cellulose), connectors (polyamide, polyvinyl chloride, rubber, silicone, Teflon), artificial veins (silicone, Dacron, Teflon), artificial lungs (polycarbonate, polypropylene, polyamide, urethane foam, polyvinyl chloride), dialyzers (Cuprophane, polypropylene, polystyrene, silicone rubber, polyvinyl chloride, nonwoven fabrics, Japanese paper), dialysis membranes (Cuprophane, polyacrylonitrile), heat exchangers (silicone rubber, stainless steel), needles (polyethylene, stainless steel, polyvinyl chloride, polyamide, rubber, Teflon), filters (polycarbonate, Dacron wool, polypropylene), and the like.
Further, the antibacterial products according to the present invention may be applied to various facilities (walls, floors, equipment, air filters, etc.) for maintaining a sterile atmosphere, endoscopes, and other things coming into direct contact with the human body.
In a cationic polyelectrolyte such as a cationic polymer, the counter ions carried thereon are generally low molecular weight counter ones (for example, halogen ions), and so is relatively easily separated from the polymer and thus the cationic sites in the polymer is easily exposed.
To the contrary, in the PEC used in the present invention, 35 2 ~ 7 , ~ i 1 the counter ions are polymeric compounds, and so the properties of quaternary a~onium from the starting cationic polymer are somewhat neutralized. It should be surprised that the PEC with such a structure exhibits antibacterial activity. The reason is not elucidated at present, but it is ass~ed tnat the quaternary ammonium portions strongly chemically bonded and contained in the polymer per se exhibit a sustained antibacterial activity.
Further, a PEC generally exhibits remarkably diverse properties along with changes in the microdomain structure having the hydrophilic property, the structural changes in the surface water, changes in the charge balance or the like, and thus, it is assumed that these effects also serve the manifestation of the antibacterial property in the PEC
of the present invention.
., Examples The present invention now will be further illustrated by, but by no means limited to, the following examples. It is noted that the average molecular weights described in the following examples are number average molecular weights measured by the vapor pressure osmometer method.
Pre~aration Exam~le 1: Pre~aration of PEC (2X-CLA) A cationic polymer, poly[(dimethyliminio)ethylene-(dimethyliminio)-methylene-1,4-phenylenemethylene dichloride] (2X) (average molecular weight = about 6000), in an amount of 0.015 9 (1 x 10-4 moles as cationic sites), and an anionic polymer, acrylic acid/lauryl acrylate random copolymer (CLA) (acrylic acid content = about 80 mole percent; average molecular weight = about 10,000), in an amount of 0.018 9 (1 x 10-4 moles as anionic sites) were dissolved separately in 10 ml of physiological saline solution (pH 7.4). The two aqueous solutions (5 ml, respectively) were mixed in a beaker to form a polyelectrolyte complex gel (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in ,, .
I, .

.
~'. ~ ' ' ' : '' ~,' ' ' . . : -, . .

-, . : . :.
,.~ - : ~ : . : , :
,,,, , ;
,, :.
:"

-36- 2~ J J
a beaker to form a polyelectrolyte complex gel ~charge balance = +4).
Further, 3 ml of the aqueous solution of the cationic polymer and 7 ~l o~ the aqueous solution of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = -4).
Pre~aration Exam~le 2: Pre~aration of PEC (2X-COA) ; A cationic polymer, poly[(dimethyliminio)ethylene-, (dimethyliminio)-methylene-1,4-phenylenemethylene dichloride] (2X~ (average molecular weight = about 3000), in an amount o~ 0.015 9 (1 x 10 4 moles as cationic sites), ' and an anionic polymer, acrylic acid/2-ethylhexyl acrylate random copolymer (COA) (acrylic acid content = about 60 molar percent; average molecular weight = about 8000), in an amount of 0.021 9 (1 x 10-4 moles as anionic sites) were ~- separately dissolved in 10 ml of distilled water (pH 8.0).
: The two aqueous solutions (5 ml, respectively) obtained `e were mlxed together in a beaker to form a polyelectrolyte ~ complex gel (charge balance = +0).
i~ The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Further, 3 ml of the aqueous solution of the cationic polymer and 7 ml of the aqueous solution of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = -4). -Pre~aration Exam~le 3~ Pre~aration of PEC (PVBMA-COA) A cationic polymer, poly(vinylbenzyltrimethylammonium chloride) (PVBMA) (average molecular weight = about 15000), in an amount of 0.021 9 (1 x 10-4 moles as cationic sites), and an anionic polymer, acrylic acid/2-ethylhexyl acrylate random copolymer (COA) (acrylic acid content = about 60 ~ mole percent; average molecular weight = about 8000), in an amount of 0.021 9 (1 x 10-4 moles as anionic sites) were ;

", ,, , ~
.... .
" , ......................... , . ,.
' , ' ,,.

-3~- 2 ~ J 3 :`~
separately disso~vtd l. 10 ~l of distilled water (pH 8.0).
The two aqutio~lj sol~ ,..s ~5 .~ espectively) obtained were rnixed -o~er;~e~ bea~er to ~orm a pol~electrolyte complex gel ~C~.~!9e ~a ' ance = tO), The aqueous solu~ion ~7 ml) of the cationic polymer and the aqueous solu- O:-l (3 ml) of the anionic polymer prepared in the sar,e ranne! as above were mixed together in a beaker to fo~m a po~;elec~!ol~te complex gel (charge balance = +4).
Further, 3 ml of he aqueous solution of the cationic polymer and 7 ml o~ s`ne aqueous solution of the anionic polymer prepa-e~ in ~ same manner as above were mixed togetner in a beaker -o _o-m a polyelectrolyte complex gel (charge balan_e = -4).
Pre~aration Exam~~le ~: ?re~aration of PEC ( PVBMA- CLA) A cationic polyme~, ~oly(vinylbenzyltri.methylammonium chloride) (~VB~A) ~ave!age molecular weight = about 100000), in an amount of 0.106 9 (5 x 10-4 moles as cationic sites), and an anionic polymer, acrylic acid/lauryl acrylate random copolymer (CLA) (acrylic acid content = about 80 mole percent; average molecular weight =
about 4000), in an amount of 0.090 9 (5 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of physiological saline solution (pH 7.4). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer .; prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~aration Exam~le 5: Preparation of PEC (2X-Sodium Alainate) A cationic polymer, poly[(dimethyliminio)ethylene-(dimethyliminio)-methylene-1,4-phenylenemethylene dichloride] (2X) (average molecular weight = about 6000), in an amount of 0.015 9 (1 x 10-4 moles as cationic sites), 2 n, ~

and a polysaccharide, s-,di~ alginate (average molecular weight = about 500,C0~,), i.-. an ,l.~;~ur.t of 0.020 9 (1 x 10-4 moles as anionic sites` were s~-~p,;lra'~eLy dissolved in 10 ml of distillec watc~ . 8, O) . '[".ee ~','10 aqueous solutions (5 ml, respec-lve'y) c~ r.~ ;er~ ed together in a beaker to for.n a ùoly- e~ c~ c~:-p e:~ ^,e (charge balance =
+O) .
Similarly, 0.075 9 of the abo~e-mentioned cationic polymer (5 x 10-4 moles as cationic sites) and 0.100 9 of the polysaccharide (5 x 10-4 m,oles as anionic sites) were separately ~issolved l~ 10 ~.l of dis.illed water (pH 8.0).
The aquesus so'_t~ n 7 .~ f t~2 cationic polymer and the aqueous solutio: ~3 ~ ~a-- ~o'~ysaccharide were mixed together in a bea~er ~ -O~- a po'yelectrolyte complex gel (charge balance = t~ ), Preparation Exa~le 6: Pre~aracion of PEC (PVBMA-Sodium Alainate) A cationic polymer, poly(vinylbenzyltrimethylammonium chloride) (PVBMA) (average molecular weight = about 15,000), in an amount of 0.106 9 (5 x 10-4 moles as cationic sites), and a polysaccharide, sodium alginate (average molecular weight = about 100,000), in an amount of 0.100 9 (5 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of distilled water (pH 8.0). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = TO), The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the polysaccharide prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Preparation Example 7: Pre~aration of PEC (2X-Polvalutamic Acid) A cationic polymer, poly[(dimethyliminio)ethylene-(dimethyliminio)-methylene-1,4-phenylene methylene dichloride] (2X) (average molecular weight = about 6000), in an amount of 0.015 9 (1 x 10-4 moles as cationic sites), -39- 2 ~
and an anioni- ~olyme~, ~,olyglu~-~mic acid (PGA) (average molecular ~,Je 9~.- = a_J;t ~000), ~n an amount of 0.013 g (1 x 10-4 moles as anion'- sites) ,J~r~ separately dissolved in 10 ml of physiological saline solut on (pH 7.4). The tr~o aqueous solutior.s (5 .~.., resrec~ve'y) obtained ~"ere mixed together in a ~ 'J ~~~rn a ?ol electrolyte co~plex gel [2X-PGA] (charge balan-~e = +Oj.
The aqueous solu~o~ (7 mlj o- the cationic polymer and the aqueous solution (3 ml) of the anionic polymer prepared in the sarne manner as aso~e were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~aration E~~le 8: 2-e~aratio~ o- PEC !pvB~A As~artic Acid/Alanine Cs~ol-,~e~`
A cationic pol~,rne-, poly(vinylbenzyltrimethylamrnonium chloride) (p~r~ ) (average molecula~ weight = about 15,000), in an amount of 0.106 9 (5 x 10-4 moles as cationic sites), and an anionic polymer, aspartic acid/alanine random copolymer [C(Asp/Ala)] (aspartic acid content = about 65 mole percent, average molecular weight =
about 8000), in an amount of 0.090 9 (5 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of distilled water (pH 9.0). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelec~rolyte complex gel [PVBMA-C(Asp/Ala)] (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of ~he anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Preparation Example 9: Pre~aration of PEC ~Polv (L-Lvsine)-CLA)l A cationic polymer, poly(L-lys ne) (PLL) (average molecular weight = about 3000), in an amount of 1.3 mg (l x 10-5 moles as cationic sites), and an anionic polymer, acrylic acid/lauryl acr~late random _opolymer (CLA) (acrylic acid content = about 80 mo e percent; average !l rJ , 7 i ! ;
molecular weig`rlt = ar~out 50GOJ, in an amount of 1.8 mg (1 x 10-5 moles as a!~ic.-ic sites) ,-ere separately dissolved in 10 ml of an aqueous solutiori of 0.5 rr;ole/liter sodium chloride (pH 6.5). 'i""e ~`.`WO aqueo-ls solutions (5 ml, respectivel~) or~a_~ d /ere ~xed ~ogether in a reaker at room temperature ~ a ~ ~e-.Fc~roi~te cornplex gel [PLL-CLA] !charge ~alance = IO) The aqueous soiution (7 ml) of the cationic polymer and the aqueous so'u~ion (3 ml) of the anionic polymer prepared in the same manner as ab~-~e were mixed ~ogether in a beaker to form a pol~electrol~te ~omplex gel (charge balance = +4).
Pre~aration Exam~le 10: Dre~aration. of PEC (Lvsine/Serine CoDolvmer-Dolvclutamic ac~d) A cationic pol;~ucr, lysine/serine random copolymer [C(Lys/Ser)] (lysine content = about 70 mole percent, average molecular weight = about 10000), in an amount of 0.019 9 (1 x 10 4 moles as cationic sites), and an anionic polymer, polyglutamic acid (PGA) (average molecular weight = about 2000), in an amount of 0.013 9 (1 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of physiological saline solution (pH 7.4). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker to form a polyelectroly~e complex gel [C(Lys/Ser)-PGA] (charge balance = -0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of ~he anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~aration Exam~le 11: Pre~aration of PEC ~Polv(L-Lvsine)-Sodium Alainate) A cationic polymer, poly(L-lys ne) (PLL) (average molecular weight = about 3000), in an amount of 1.3 mg (1 x 10-5 moles as cationic sites), and an anionic polymer, sodium alginate (Arg) (average molecular weight = about 40000), in an amount of 1.8 mg (1 x 10-5 moles as anionic sites) were separately dissolved in 10 ml of an aqueous -41- ~ 97,,1~3 ~
solution of 0.5 mole/liter sodium chloride (pH 6.5). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelectrolyte complex gel [PLL-Arg] (charge balance =
+0) -The aqueous solution (7 ml~ of the cationic pol~mer and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~a ation Example 12: Pre~aration of_PEC (Lysine/Serine Co~olvmer-Carboxvmethvlchi~in~
A cationic polymer, lysine/serine [C(lys/Ser)] random copolymer (lysine conten~ = about 70 mole percent) (average molecular weight = a~ou~ 10000), in an amount of 0.019 9 (1 x 10-4 moles as cationic sites), and an anionic polymer, carboxymethylchitin (CM-Chn) (carboxymethylation degree =
about 0.65/monosaccharide) (average molecular weight =
about 5000), in an amount of 0.018 9 (1 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of a physiological saline solution (pH 7.4). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelectrolyte complex gel [C(Lys/Ser)-CM-Chn] (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer , s prepared in the same manner as above were mixed together in .~ a beaker to form a polyelectrolyte complex gel (charge .~s, balance = +4).
,:
Preparation Exam~le 13: Pre~aration of PEC (Chitosan-Sodium ' Alainate) A cationic polysaccharide, chitosan (deacetylization . degree = 100 percent; average molecular weight = about .;s 2000), in an amount of 0.020 9 (1 x 10-4 moles as cationic - sites), and an anionic polymer, sodium alginate (Arg) ~ (average molecular weight = about 4000), in an amount of 0.018 g (1 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of a physiological saline solution (pH

. . .

:, ~ - , .
- : .
, , ., .,. . , - ~:, :

~:.. - , ~ . :
;'5 ' ' ' ' .'. , , ,~,~- . .....
,,' ' ,, ' ' :' :', . , . ': ' . .~' , 2 ~ ~ r 7.4). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelectrolyte complex gel (chitosan-Arg) (charge balance = +0).
The aqueous solution (7 ml) of the cationic polysaccharide and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Preparation Exam~le 14: Pre~aration of PEC (Chitosan-Sulfated Cellulose) A cationic polysaccharide, chitosan (deacetylization degree = about 70 percent; average molecular weight = about 5000), in an amount of 0. 102 9 ( 5 x 10 4 moles as cationic sites), and an anionic polymer, sulfated cellulose (S-cel) (sulfation degree = about 0.8/monosaccharide; average molecular weight = about 8000), in an amount of 0.110 9 (5 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of distilled water (pH 5.0). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelectrolyte complex gel (chitosan-S-cel) (charge balance = +0).
The aqueous solution (7 ml) of the cationic polysaccharide and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~aration Exam~le 15: Pre~aration of PEC
(Diethvlaminoethyldextran-Carboxvmethvlchitin) A cationic polysaccharide, diethylaminoethyldextran (DEAE-Dex) (rate of introduction = 60 percent; average molecular weight = about 3000), in an amount of 2.0 mg (1 x 10-5 moles as cationic sites), and an anionic polymer, carboxymethylchitin (CM-Chn) (carboxymethylation degree =
about 0.65/monosaccharide; average molecular weight = about 5000), in an amount of 1.8 mg (i x ~0-5 moles as anionic sites) were separately dissolved in 10 ml of an aqueous solution of 0.5 mole/liter sodium chloride (pH 8.0). The .'.'' ~

. .

-, . ,, . , - :, .
, ~ . , : - :
; . : :.

,; .

- 4 ~
two aqueous solutlor.s (. rnl, res?fctivel~) obtained were mixed together i~ a _e~er a~- ~oo~" temperature to form a polyelectrol~e cornr)le~ gel 3~ .De~-CM-Chn) (charge balance = ~o).
The aq~eous soi~r o.~ (7 ~ of the cationic polysaccharide ard t.~e a~le^l~s solution (3 ml) of the anionic pol-~m~er prepared in the same manner as above were mixed together i~ a bea.ker to form a polyelectrolyte complex gel (charge balance = +4).
Pre~aration Exam~le '.6: ?re~ara~ion of PEC (Chitosan-Polvalutamic ..cid) ~ . cationic pol,~,ie-, chi ~^san (deacetylization degree =
100 percent) (average molecular ~;eight = about 2000), in an amount of 0.020 9 (1 x 10-4 moles as cationic sites), and an anionic polymer, polyglutamic acid (PGA) (average molecular weight = about 4000), in an amount of 0.013 g (1 x 10 4 moles as anionic sitesj were separately dissolved in 10 ml of a physiological saline solution (pH 7.4). The two aqueous solutions (5 ml, respectively) obtained were mixed together in a beaker at room temperature to form a polyelectrolyte complex gel (Chitosan-PGA) (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer prepared in the same manner as above were mixed together in a beaker to form a polyelectrolyte complex gel (charge balance = +4).
Preparation Exam~le 17: Preparation of PEC (Chitosan-CLA) A cationic polymer, chitosan (deacetylization degree =
70 percent) (average molecular weight = about 3000), in an amount of 0.021 g (1 x 10-4 moles as cationic sites), and an anionic polymer, acrylic acid/lauryl acrylate random copolymer (CLA) (acrylic acid content = about 80 molar percent) (average molecular weight = about 4000), in an amount of 0.018 9 (1 x 10-4 moles as anionic sites) were separately dissolved in 10 ml of an aqueous solution of 0.3 mole/liter sodium chloride (pH 6.5). The two aqueous solutions (5 ml, respectively) obtained were mixed together 2~7 ~
. .~
in a beaker at room ~em?eratllre to form a polyelectrolyte complex gel (Cnitosan-C~ (charge balance = +0).
The aqueous solution (7 ml) of the cationic polymer and the aqueous solution (3 ml) of the anionic polymer prepared in ~he scu`,e mar...e- as abo;/e were mixed together in a beaker -o orm. a ~J'.-el--c~ e complex gel (charge balance = +4).
Immobilization Ecam~le '.: Tr~m.ooillzation to Carrier (Polvethvlene Tube~
Two ml amounts of ~he DEC gels prepared in Preparation Examples 1 to 17 were pou-ed into polyethylene tubes (inside diamecer = ~.C ~` and ro~a~ed by a rotor at 60 rpm for 8 hours, res~ec~ e~~ er the PEC was coated on the inside walls of the DO' `,'e-- ;l ene tubes, the supernatant was removed. The tubes ~,~ere ~-ied at 60C for 4 hours, washed the insides by 10 ml of distilled water three times, and then were again dried at 80C for 4 hours to obtain the PEC
immobilized tubes.
Imrnobilization Example 2: Immobilization to Carrier (Gauze) Pieces of cotton gauze (10 cm x 10 cm) were immersed for 8 hours in the PEC gel liquids prepared in Preparation Examples l to 17. The gauze pieces were taken out therefrom, washed with 20 ml of distilled water, dried at 60C for 4 hours, then washed by 10 ml of distilled water three times, and dried a~ 80C for 4 hours to obtain the PEC immobilized gauze.
Immobilization Exam~le 3: Immobilization to Carrier (Glass Beads) Glass beads (diameter = 0.2 mm; made by Toshiba Varotini K.K.) were immersed for 6 hours in the PEC gel liquids prepared in Preparation Examples l to 6. After the glass beads were coated by the PEC, the supernatant was removed. The beads were dried at 60C for 4 hours, washed by 10 ml of distilled water three times, and again dried at 80C for 4 hours to obtain PEC immobilized beads.
Immobilization Exam~le 4: Immobilization to Carrier ~Contact Lens Material (Dolvmethvlme~hacrvlate)l 2 " 7 ~

- Small circular pieces (diameter = 8 mm) punched out from polymethylmethacrylate sheets made of the material same as that for con~act len.ses (hereinafter referred to as "small circular pieces") ~ere immersed at room temperature for 10 hours in the PEC gel liquids prepared in Preparation Examples 7 to 17. Afte~ .e small circular pieces were coated by the PEC, the supernatant was removed. The pieces were dried at 60C for 3 :sours, washed by 10 ml of distilled water twice and washed by physiological saline solutions twice, and the~ dried at 80C for 4 hours to obtain PEC immobilized small circular pieces.
Pharmacoloqical Test Exam~le 1: Examination of Antibacterial Pro~er v ,:, The following microo-ganisms were used to examine the antibacterial property.
-. Escherichia coli ATCC25932 ,~ Staphylococcus aureus ATCC25923 :~ Serratia marcescens IFO3046 - Pseudomonas aeruginosa ATCC10145 " ~
The various bacterial solutions cultivated in brain heart infùsion (BHI) media at 37C for 16 hours were diluted by an M/15 phosphate buffer solution (PBS, pH 6.8) containing 0.85 % sodium chloride to prepare bacterial suspensions having a bacteria concentration of about 1 x 104/ml. After 10 ml of a PD medium ~prepared by dissolving 7.0 9 of dipotassium hydrogenphosphate, 2.0 9 of potassium hydrogenphosphate, 0.1 9 of magnesium sulfate, 1.0 9 of ammonium sulfate, 0.5 9 of sodium citrate, 10.0 9 of glucose and 10.0 9 of bactopeptone in 1000 ml of purified water) was added to the PEC-coated polyethylene tubes prepared in the above Immobilization Example 1, 0.1 ml of each of the above bacterial suspensions was added. The whole was mixed, and then held at 30C. Shaking cultivation was carried out at 30C for 24 hours and the turbidity of the media was visually observed to examine the antibacterial effect. The results are shown in Table 1.
Pharmacoloaical Test Exam~le 2: Examination of Antibacterial Pro~ertv ' ,- : ' ' ~ , ' ..

:

; ............ . .

."
, - ..

, .

-~6-Fifty ml portions of a P~ ~,ediwn were inserted into Sakaguchi-flas'~s, the?. ~;~ree plr:'_S of the PEC-immobilized gauze (6 cm x 6 cn~ o's~-~ined in rmrnobilization Example 2 were introduced to the -'~asks, ard 1 ml o~ the bacterial suspensions used in the above Pharmacological Test Example 1 were inoculated ~r. ar. amourlt of about 1 x 10~ bacteria.
Further, inoculation was carried out in the same manner as above, in each of flas'~s for a blan'.~- test and those for a control test wherein three pieces of gauze (6 cm x 6 cm) without immobilize-D~C r.~ere in~rodu^ed. Then, shaking cultivation was carrie~ OUb at 35~ -or 16 hours for each flask. The turbid ti o- the meclia :;as visuall~ observed to examine the antibacteri~' effect. ~'ne results are shown in Table 2.
Pharmacoloaical Test Exam~le 3: Exa~ination of Antibacterial Pro?ertv Ten ml portions of a PD medium were poured into sterilized polyethylene tubes, then 1 9 of the PEC-immobilized glass beads obtained in Immobilization Example 3 was introduced thereto and 0.1 ml of the bacterial suspensions as in the above-mentioned Pharmacological Test Example 1 was added. The whole was mixed and held at 30C.
Further, inoculation was carried out in the same manner as above, in each of tubes for a blank test and those for a control test wherein 1 9 of glass beads without immobilized-PEC was introduced. Then, shaking cultivation was carried out at 30C for 24 hours for each tube. The turbidity of the media was visually observed to examine the antibacterial effect. The results are shown in Table 3.
Pharmacoloaical Test Exam~le 4: Examination of Antibacterial Pro~ertv Ten ml portions of a PD medium were poured into sterilized polyethylene tubes, then 2 9 of the PEC-immobilized small circular pieces obtained in Immobilization Example 4 was added and 0.1 ml of the bacterial suspensions as in the above Pharmacological Test Example 1 was added. The whole was mixed and held at 30C.
Further, inoculation was carried out in the same manner as 2,, J

above, in each of tubes for a biank test and those for a control test wherein 2 g of the small circular pieces without immobilized-?EC ',`/a5 introduced. Then, shaking cultivation was carried out at 30C for 24 hours for each tube. The turDidltv of ~.e media was visually observed to examine tne antibacreri--l efrecc. .ne results are s;~own in Table 4.
Pharmacoloaical Test E~a.nole 5: Exa~ination of Sustained Antibacterial Prooertv Ten ml portions Ot purified wa-er were added to PEC-immobilized tubes ore?ared in I~moD lization Examole 1, then the tubes were ~iy^~olisl,~ s~ake~ in a shaker for 2 minutes, and the washir.y solutions were discarded. The washing as above was reoeated furthe- four times !five times in total), and then the ~ubes were dried at 60C for 2 hours. The thus pre-treated tubes were used to examine the sustained antibacterial effect D'~ the procedure as in Pharmacological Test Example 1 for E. coli (ATCC25932).
The results are shown in Table 5.
Pharmacoloaical Test Example 6: Examination of Sustained Antibacterial ProPertv - Pieces of PEC-immobilized gauze prepared in Immobilization Example 2 were introduced in 1000 ml beakers, 500 ml of purified water was added thereto, magnetic stirrers were used for agitation for 5 minutes, and then the washing solutions were discarded. The washing . as above was repeated further four times (five times in total). Then, the pieces of gauze were taken out therefrom and dried at 60C for 4 hours. The thus pre-treated pieces of gauze were used to examine the sustained antibacterial effect by the procedure as in Pharmacological Test Example 2 for E. coli (ATCC25932). The results are shown in Table 6.
Pharmacoloaical Test Example 7: Examination of Sustained Antibacterial Propertv PEC-immobilized glass beads prepared in Immobilization Example 3 were placed in sterilized ?olyethylene tubes, 10 ml of purified water was added, then the tubes were ~;?, 5 / r vigorously s~aken in a s;~a':er for 2 minutes, and then the ~ashing solutions .:e~e I s^arae~. The washing as above was repeated further four ~imes (f~;e times in total), then the pieces of gauze were ta~en. out therefrom and were dried at 60C for 4 hours. ~r~.e thus pre-treaced tubes were used to examine the sustaineà as i~acterial eflect by the procedure as in Pharmacological Test ExamDle 3 for ~. coli (ATCC25932). The ~esllt~ are shown ln Table 7.
Pharmacoloaical Test Exa~nDle 8: E~a~ination of Sustained Antibacterial ProDerc~
PEC-immobilized small circular pieces prepared in Immobilization Example ~ /e-e intrcduced into sterilized polyethylene tibes, 19 rnl of p.lrifi-d water was added, then the tubes were vigorousl;~ sha'~en in a shaker for 2 minutes, tnen the washing solutiona ~ere discarded. The ~Jashing as above was repeated further four times (five times in total), then the small circular pieces were taken out therefrom and dried at 60C for 4 hours. The thus pre-treated small circular pieces were used to examine the sustained antibacterial effect by the procedure as in Pharmacological Test Example 4 for E. coli (ATCC25932).
The results are shown in Table 8.
Pha_macoloaical Test Exam~le 9: Examination of Antibacterial Pro~ertv Pieces of filter pa?er (TOYO, "o. 5B) were immersed for 8 hours in the PEC gel solutions prepared in Preparation Examples l to 17. The ~ieces of filter paper were taken out therefrom and washed ,with 20 ml of distilled water, then dried at 60C for 4 hours, further washed by l0 ml of distilled water three times, and dried at 80C for 4 hours. The resulting filter paper pieces were cut into circular pieces having a diameter of 13 mm and subjected to gas sterilization treatment to obta n the test disks for antibacterial effect as below. Fur her, the same procedure was repeated, except that the circu_ar filter paper not immersed in PEC gel solutions was used, to prepare control disks.

~ I ~ r~

"
The our t~oes of -~cserla described in Pharmacological Tes~ 51~ 1 ;ere shake-cultivated overnight in B:~I medla ;3~t ~he same method as in Pharmacological Test Example l and subjected to three centrifugation treatments b~ the ~HI media, then diluted by BHI media to p.epare bac~e~ial sus?ensions having a bacteria concentration of about 1 x 107/ml. Twenty ~l portions of the bacterial suspen.sions were inoculated in the above antibacterial effecs test disks and control disks. The disks were allowed to scand at 37C ~or 2 hours, then placed on Tr~Dto-so~a agar plates so that the inoculated sides of ~he d sks c~ntacted the agar plates.
The whole was allo,weà to sr~nd at 37C for 1 hour, then the disks were removed. The p~a~es were incubated at 37C
overnight, then the forma~ion of colonies on the plate was observed. The results are shown in Table 9.
Examples 1 to 11 Various types of PEC were prepared in the same manner as in the above-mentioned preparation examples and immobilized on carriers, and then, the pharmacological activities were observed. More particularly, the cationic polymers shown in Table 10 and the anionic polymers shown in Table 11 were dissolved in the amounts shown in the "Amount taken" columns of Tables 10 and 11 in 10 ml of the solvents shown in the "Solvent" column of Table 10. The obtained solutions (5 ml, respectively) were mixed all at once in a beaker to form a polyelectrolyte complex gel (charge balance = +0). Similarly, polyelectrolyte complex gels having a charge balance of +4 or -4 were formed from 7 ml of the cationic polymer solutions and 3 ml of the anlonic polymer solutions, or from 7 ml of the anionic polymer solutions and 3 ml of the cationic polymer solutions. The resulting PEC gels were immobilized on carriers (gauze) by the same method as in Immobilization Example 2 and the antibacterial properties were observed by the same method as in Pharmacological Test Example 2. The results are shown in Table 12. It is noted that in Tables 1~ and 11, the polymers are shown by abbreviations. The meanings of the abb.eviations are as follows (the abbreviations used in the above Preparation Examples have the same meanings, so ~.e ex?lanations thereabout are omitted~.
6X: Poly[(dimethyliminio)hexamethylene(dimethyliminio)-methylene-1,4-phenyler.emethylene dichloride]
PAA: Polyacrylic acid PSS: Polystyrenesulfcnic acid SLA65: Styrenesulfonic acid/laurylacrylate random copolymer (styrene sulfonic acid content = about 65 mole percent) CSA74: Acrylic acid/s~earylacrylate random copolymer (acrylic ac d con~en~ = about 74 mole percent) CLA66: Acrylic acid/laury,acrylate random copolymer (acrylic acid content = about 66 mole percent) QPAl Am: Quaternized polyallylamine In the following Tables 1 to 8 and 12, the symbols have the following meanings:
+++: strong turbidity ++: turbidity +: a little turbidity +: no change -: transparent.

Table 1 PEC Charge E. S. S.marce- P. aerugi-balance coli aureus scens nosa 2X-CLA +0 - - - +
+4 -4 ++ ++ +++ +++
2X-COA +0 + ~+ +++ +++
+4 - - +
-4 ++ ++ +++ +++

~1 t~ r PVBl`qA- +O - - -t+
COA +4 - - + ++
-4 +i- +-t +-t-t +-t+
_ _ . _ _ PVBMA- +O + T -t -t +-t CLA ~4 -t~ ++
2X-alginlc 'O
acid +4 -PVBMA- +O - - + +
alginic +4 1 - ++ ++
acid _ _ 2X-PGA tO + T + +
+4 + +
PVBMA- +O + T + +
C(Asp/Ala) - 4 - - + +
PLL-CLA +0 - - - -+4 C(Lys/Ser)- +O + ++ ++ ++
PGA +4 + - - -PLL-Arg +0 - _ ~ +
+4 - - +
C(Lys/S~r) _O + + + +
-CM-chn +4 - -~ - +
Chitosan- _0 + + + +
Arg +4 + +
_ _ Chitosan- +0 + + + +
_ S.c~1 +4 DEAE Dex- +0 CM-Chn +4 -~2- ~7 -'?~`
Chitosan- 3 ~ + -t PGA +4 - - - +
~ = = = = . . . _ _ Chitosan- t O t -t + +
CLA +4 - - T +
-Blank test +r -t-~ +-t-t +++

Table 2 PEC Charge E S. S. marce- P. aerugi-balance coli aureus scens nosa +4 -4 ++ T+ +++ +++
2X-COA +0 ++ T+ ++ ++
+4 +
-4 ++ ++ +++ +++
PVBMA- +O - - ++ ++
COA +4 + + ++ +++
-4 + +
PVBMA- _O + + +++ +++
CLA +4 + ++ +++ +
2X-alginic +O
acid +4 PV3MA- +O - - + ++
alginic +4 + T ++ ++
acid 2X-PGA +0 + + + +
+4 - + +
PVBMA- +O + + + +
C(Asp/Ala) +4 - - T

, -53- ~', 7 `, ~
PLL-CLA O
+4 C ~Lys/Ser) - +O -' +-~ +~~ ++
PGA + 4 PLL-Arg +O - + _ +
+ ~ +

C (Lys/Ser) - +O + + t +
CM - chn +4 - T - +
Chitosan- 10 t t -~ +
Arg +4 - - + +
Chitosan- +O + + + +
S.cel +4 DEAE-Dex- +O
CM-Chn +4 - - - - -Chitosan- +O + + + +
PGA +4 - - - +
Chitosan- +O + + + +
CLA +4 ~ ~ +
Blank test ++ ++ +++ +++
Gauze alone ++ ++ +++ +++

Table 3 PEC Charge E. S. S. marce- P. aerugi-balance coli aureus scens nosa 2X-CLA +O - - + +
+4 -4 ++ ++ +++ +++

.5~

.~
2X-COA +0 +t -t~ +t +4 -4 ++ ++ +t-~ -t-~+
PVBMA- +0 - - t+ -t+
COA +4 - ++ --t-t ++ + ++
PVBMA- +0 ~ + +++ +++
CLA +4 + ++ ~++ +t+

2X - alginic +O
acid +4 PVBMA - + O
alginic +4 + ~ ++ ++
acid Blank test ++ ++ +++ +++
Beads alone ++ ++ +++ +++

Table 4 PEC Charge E, S. S. marce- P. aerugi-balance coli aureus scens nosa 2X-PGA +0 + + + +
+4 - + +
~, PVB~A - + O + + + +
C (Asp/Ala) +4 - - + +
PLL-CLA +0 - - - -+4 C (Lys/Ser) - +O + ++ ++ ++
. PGA +9 +
PLL-Arg +0 - + + +
+4 - - +

' ~

C (Lys/Ser) +O -~ + -t +
C~ chn +4 - ~ - +
_ __. __ __ Chitosan- +0 + + + +
Arg +4 - - + +
Chitosan- +0 + -~ + +
S.cel +4 - - - -DEAE Dex- +0 CM-Chn +4 Chitosan- +0+ + + +
PGA +4 - - - +
Chitosan- +0t + + +
CLA +4 - - + +
Blank test ++ ++ +++ +++
Small circular pieces alone ++ ++ +++ +++

Table 5 PEC Charge balance E. coli 2X-CLA +0 +4 -4 ++
2X-COA +0 +
+4 -4 ++
PVBMA-COA +O
+4 -4 ++
PVBMA-CLA +O +
+4 ++

-5~- 2;.~ J~.;
2X-alginic acid ~0 ~ .~
PVBMA-alginic ~0 acid +4 2X-PGA +O
+4 PVBMA- +O
C(Asp/Ala) +4 PLL-CLA ~0 +4 C(Lys/Ser)- +O
PGA +4 PLL-Arg _O
+4 C(Lys/Ser)- +O
CM-chn +4 Chitosan- +O
Arg +4 Chitosan- +O
S.cel +4 DEAE Dex- +O
CM-Chn +4 Chitosan- +O
PGA +4 ....
Chitosan- +0 +
CLA +4 Blank test ++

' , .

Table 6 PEC Charge balance E. coli 2X-CLA _0 t4 - 4 t-r 2X-COA tO ++
+4 -4 +T
PVBMA-COA t O
+4 ~4 t PVBMA-CLA +0 +
+4 +
2X-alginic acid +0 +4 PVBMA-alginlc +0 acid +4 +
2X-PGA +0 +
+4 PVBMA- +o +
C(Asp/Ala) +4 PLL-CLA +0 +4 C(Lys/Ser)- +0 +
PGA +4 +
PLL-Arg _0 +4 C(Lys/Ser)- +0 +
CM-chn +4 --5~ '.;J 3 ~
Chitosan- +G +
Arg t4 Chitosan- -0 +
S.cel +~1 -DEAE Dex- ~0 C~l-Ch~ +~1 Chitosan- _0 +
PGA +41 Chitosan- _0 -, CLA r~ -Blank test ++

Gauze alone ++
. .
Table 7 PEC Charge balance E. coli ~ 2X-CLA +0 :~ +4 -4 +~
- 2X-COA +0 ++
+4 -4 ++
: PVBMA-COA +0 :~ +4 +
-4 +
PVBMA-CLA +O +
+4 +
2X-alginic acid +0 +4 , ,~
: ~ ' ' ' ~ :

.

PVBMA-alginlc -O
acid --4 Blank test -~
Beads alone ;+

Table 8 PEC Charge balance E. coli 2X-PGA +O -~
+4 PVBMA- +O
C(Asp/Ala) +4 PLL CLA +O
+4 C(Lys/Ser)- +O +
PGA +4 PLL-Arg +O
+4 C(Lys/Ser)- +O +
CM-chn +4 Chitosan- +O +
Arg +4 Chitosan- +O
S.cel +4 DEAE Dex- +O
CM-Chn +4 Chitosan- +O +
PGA +4 , F r Chitosan- tO +
CLA ~4 _ _ Blank test -t t ___~__ _ _ _ Small circular pieces alone -t-+

In the follo,ing Table 9, the s~ols show the degree of colony format on. The degrees are as --ollc;s +++: Great ++: Medium +: Small +: Extremely sma' -: No change Table 9 PEC ChargeE. S. S. marce- P. aerugi-balance coli aureus scens nosa 2X-CLA +0 - - + ++
:: +4 - +
-4 ++ ++ +++ ++
2X-COA +0++ ++ ++ ++
+4 +
-4 ++ ++
PVBMA- +0 - - ++ ++
COA +4 + + + ++
-4 + + ++ +++
PVBMA- +0 + + ++T ++
CLA +4 + ++ +++
2X-alginic +0 - - - +
acid +4 ~ ~ ~ +

, ~J - i ,`. ' .1 ~

PVBMA- +O - - + -t+
alginic +4 ~ - - + t acid 2X-PGA +O -t -r t +
+4 PVLMA- +O + - +
C(Asp/Ala) +4 - - + +
PLL-CLA +O
+4 _ C(Lys/Ser)- +O + - t PGA + 4 PLL-Arg +O - / + +
+4 - - - t C(Lys/Ser)- +O + + + +
cM-chn +4 +
Chitosan- +O + + + +
Arg +4 - - - +
Chitosan- +O + + + +
S.cel +4 DEAE Dex- +O - - + +
CM-Chn +4 Chitosan- +O + + + +
PGA +4 - - - +
Chitosan- +O + + + +
CLA +4 + +
Control +++ +++ +++ +++

Table 10 Ex. Cationic Average ~nount Cationic Solvent pol~ner ~olecular taken sites weight (g) (rnoles) 1 6X 12000 0.02C 1 x 10 4 Distilled water (pH 8.5) 2 2X 80G0 0.007 5 x 10-5 Distilled water (pH 9.0) 3 2X 20000 O.G015 1 x 10-5 O.5M sodium chloride solution (pH 5.0) 4 2X 6000 0.015 1 x 10-4 Distilled water (pH 7.0) 6X 12000 0.201 1 x 10-3 0.2M sodium chloride solution (pH 6.5) 6 ~PAl.Am 13600 0.013 1 x 10-4 Distilled water (pH 8.0) 7 2X 6000 0.015 1 x 10-4 Distilled water . (pH 6.0) 8 6X 6000 0.020 1 x 10-4 0.8M sodium chloride solution (pH 4.5) 9 6X 10000 0.010 5 x 10-5 Distilled water ~ (pH 10.0) : 10 6X 10000 0.020 1 x 10-4 Distilled water (pH 9.0) 11 Chitosan 4000 0.008 5 x 10-5 Distilled water (pH 7.2) ' Table 11 Ex. Anionic Average Amount Anionic sites polymer molecular taken (g) (moles) weight 1 CLA66 15000 0.021 1 X 10-4 -~3~ .. ~ r -~
2 PAA 35000 0.005 5 X 10-5 3 PSS 40000 0.0021 1 X 10-5 4 Phosphated 7000 0.032 1 x 10-4 chitin Chondroitin 23000 0.460 1 x 10-3 sulfuric acid type A
6 Sodium 500000 0. 040 1 x 10-4 alginate 7 Phosphated 23000 0.029 1 x 10 ~4 cellulose 8 SLA65 15000 0.031 1 x 10 ~4 9 CSA74 10800 0.011 5 x 10-5 Carboxymethy- 22000 0.022 1 x 10 ~4 cellulose (substitution degree 100) 11 Carboxymethyl 23000 0.012 5 x 10-5 cellulose (substitution degree 90) Table 12 PEC Charge E. S. S. marce- P. aerugi-balance coli aureus scens nosa 6X-C~A66 -4 + + + +
+O
+4 2X-PPA - 4 + + ++ ++
+O
+4 2X-PSS 0 ++ ++ +++ +++
+4 + + + +

2X-phos- -4 r -~ +-~- +~
phated +o t r + +
chitin +4 6X-chondo- -4 -t + ++-~ ~++
roitin +O - - -r+ +
sulfuric +4 - - + +
acid type A
QPAl Am- -4 + -r sodium +O - - r +
alginate +4 2x- -4 + ~ +-r ++
phosphated +O
callulose +4 6X-SLA65 +0 + + ++ +++
+4 + + + +
6X-CSA74 -4 + + + +
+ O
+4 , 6X-carboxy - 4 + ++ ++ ++
methyl- +O - +
cellulose +4 (substitution degree 100) .:
Chitosan- -4 + + +++ +++
carboxy- +O - + + +
methyl- +4 cellulose (substitution ; degree 90) Control ++ ++ +++ +++

I~USTRIAL APPLIC~BILITY
The antibacterial agent according to the present invention can be applied to a wide range usage, because of insolubility in general solvents. Further, the antibacterial activity can be maintained for a long period of time. Still further, various antibacterial agents exhibiting various antibacterial strengths can be easil~
provided.

Claims (11)

1. An antibacterial agent characterized by containing a polyelectrolyte complex prepared by reacting a cationic polymer containing N+ atoms in repeating units thereof and an anionic polymer containing -COO-, -SO3-, or -PO3- groups in repeating units thereof.

2. An antibacterial agent according to claim 1, wherein a concentration ratio of cationic sites of said cationic polymer and anionic sites o said anionic polymer (cationic sites/anionic sites) is 0.25 to 4Ø

3. An antibacterial agent according to claim 1, wherein at last one compound selected from the group consisting of:
(a1) a compound of the general formula (I):
(I) wherein R1 and R4 are, independently, an alkylene group of 1 to 10 carbon atoms, a group of the general formula:

wherein R11 and R12 are, independently, an alkylene group of 1 or 2 carbon atoms, or an arylene group, and R2, R3, R5, and R6 are, independently, an alkyl group of 1 to 3 carbon atoms, or R1 forms, together with the 2 nitrogen atoms and R2, R3, R5 and R6 in the formula, a group of the formula:
R4 has the same meaning as above, X1- is a counter ion, and m is a number of not less than 5, (a2) a compound of the general formula (II):

(II) wherein A is a group of the general formula:

wherein B is an alkylene group of 1 or 2 carbon atoms, R21, R22 and R23 are, independently, a hydrogen atom or an alkyl group of 1 to 3 carbon atoms, and X2- is a counter ion, or A is a group of the general formula:

wherein R24 is an alkyl group of 1 to 3 carbon atoms or a benzyl group and X3- is a counter ion, or A is a group of the general formula:
wherein R25 is an alkyl group of 1 or 2 carbon atoms, R26, R27 and R28 are, independently, a hydrogen atom or an alkyl group of 1 to 3 carbon atoms, and X4- is a counter ion, and n is a number of not less than 10, (a3) a compound of the general formula (III):
(III) wherein v is 3 or 4, R91 is a hydrogen atom; an alkyl group of 1 to 4 carbon atoms; an alkyl group of 1 to 4 carbon atoms substituted by a hydroxyl or mercapto group, or by an alkylthio group of 1 to 3 carbon atoms; or an imidazolylmethyl or indolylmethyl group; R92 is -N+H2X5- or -N+HC(NH)NH2X6-; X5- and X6- are, independently, counter ions, t is 20 to 100, and r is an integer of not less than 10, and (a4) a cationic polysaccharide, is used as said cationic polymer

4. An antibacterial agent according co claim 1, wherein at least one compound selected from the group consisting of:
(b1) a compound of the general formula (IV):

(IV) wherein u is 1 or 2, R81 is a hydrogen atom, an alkyl group of 1 to 4 carbon atoms; an alkyl group of 1 to 4 carbon atoms substituted by a hydroxyl or mercapto group, or by alkylthio group of 1 to 3 carbon atoms; or an imidazolyl-methyl or indolylmethyl group; s is 20 to 100 and q is an integer of not less than 10, (b2) a compound of the general formula (V):

(V) wherein R31 is a hydrogen atom or a methyl group, R32 is an alkyl group of 6 to 18 carbon atoms, Y is a group of a carboxlic acid or a salt thereof, a sulfonic acid or a salt thereof or a phosphoric acid or a salt thereof, or an aryl group containing a group of a carboxylic acid or a salt thereof, a sulfonic acid or a salt thereof or a phosphoric acid or a salt thereof, a is 20 to 100, and p is an integer of not less than 10, and (b3) an anionic polysaccharide, is used as said anionic polymer.

5. An antibacterial agent according to claim 3, wherein a compound (a1) of the general formula (I) wherein R1 and R4 are, independently, a straight or branched alkylene group of 2 to 8 carbon atoms, and R11 and R12 are bonded at the p-position, is used.

6. An antibacterial agent according to claim 3, wherein a compound (a3) of the general formula (III) wherein R91 is a hydrogen atom, or a methyl, isopropyl, isobutyl, s-butyl, hydroxymethyl, hydroxyethyl, methylthioethyl, mercaptomethyl, 5-imidazolylmethyl or 3-imidazolylmethyl group, is used.

7. An antibacterial agent according to claim 3, wherein chitosan, a chitosan derivative, or a neutral polysaccharide diethylaminoethyl derivative is used as said cationic polysaccharide (a4).

8. An antibacterial agent according to claim 4, wherein the compound (b1) of the general formula (IV), wherein R91 is a hydrogen atom, or a methyl, isopropyl, isobutyl, s-butyl, hydroxymethyl, hydroxyethyl, methylthioethyl, mercaptomethyl, 5-imidazolylmethyl or 3-imidazolylmethyl group, is used.

9. An antibacterial agent according to claim 4, wherein the compound (b2) of the general formula (V) wherein R32 is a straight or branched alkyl group of 6 to 14 carbon atoms, and a is 50 to 100, is used.

10. An antibacterial agent according to claim 4, wherein at least one anionic polysaccharide (b3) selected from the group consisting of hyaluronic acid or a derivative thereof, alginic acid or a derivative thereof, chondroitin sulfuric acid A, chondroitin sulfuric acid C, chondroitin sulfuric acid B (dermatan sulfuric acid) or a derivative thereof, chondroitin sulfuric acid D or a derivative thereof, chondroitin sulfuric acid E or a derivative thereof, heparan sulfuric acid or a derivative thereof, heparin or a derivative thereof, K-carragheenan or a derivative thereof, .lambda.-carragheenan or a derivative thereof, a carboxymethylated, sulfated or phosphated cellulose derivative, a carboxymethylated, sulfated or phosphated chitin derivative, carboxymethylstarch or a derivative thereof, a carboxymethylated, sulfated or phosphated amylose derivative, a carboxymethylated, sulfated or phosphated amylopectin derivative, a carboxymethylated, sulfated or phosphated .beta.-1,3'-glucan derivative, a carboxymethylated, sulfated or phosphated .beta.-1,2'-glucan derivative, a carboxymethylated, sulfated or phosphated .beta.-1,3'-; .beta.-1,6'-glucan derivative, a carboxymethylated, sulfated or phosphated dextran derivative, a carboxymethylated, sulfated or phosphated pullulan derivative, a carboxymethylated, sulfated or phosphated agarose derivative, a carboxymethylated, sulfated or phosphated .beta.-1,4'-galactan derivative, a carboxymethylated, sulfated or phosphated mannan derivative, and a carboxymethylated, sulfated or phosphated inulin derivative, is used as said anionic polysaccharide.

11. An antibacterial material characterized by carrying a polyelectrolyte complex according to any one of claims 1 to 10 on a carrier.