U.S. patent application number 10/868195 was filed with the patent office on 2007-09-13 for use of bacterial phage associated lysing enzymes for the prophylactic and therapeutic treatment of various illnesses.
Invention is credited to Vincent Fischetti, Lawrence Loomis.
Application Number | 20070212340 10/868195 |
Document ID | / |
Family ID | 32469737 |
Filed Date | 2007-09-13 |
United States Patent
Application |
20070212340 |
Kind Code |
A1 |
Fischetti; Vincent ; et
al. |
September 13, 2007 |
Use of bacterial phage associated lysing enzymes for the
prophylactic and therapeutic treatment of various illnesses
Abstract
A method for the prophylactic and therapeutic treatment of
bacterial infections of the skin is disclosed which comprises the
treatment of an individual with an effective amount of a native or
altered version of a lytic enzyme produced by a bacteria infected
with a bacteriophage specific for said bacteria. The lytic enzyme
is selected from a group consisting of shuffled lytic enzymes,
chimeric lytic enzymes, wherein the lytic enzyme is in an
environment having a pH which allows for activity of said lytic
enzyme; and a carrier for delivering said lytic enzyme.
Additionally, a holin protein for puncturing the membrane may be
included in the composition.
Inventors: |
Fischetti; Vincent; (West
Hempstead, NY) ; Loomis; Lawrence; (Columbia,
MD) |
Correspondence
Address: |
Jonathan E. Grant
2107 Hounds Run Place
Silver Spring
MD
20906
US
|
Family ID: |
32469737 |
Appl. No.: |
10/868195 |
Filed: |
June 16, 2004 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09560650 |
Apr 28, 2000 |
6752988 |
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10868195 |
Jun 16, 2004 |
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09395636 |
Sep 14, 1999 |
6056954 |
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09560650 |
Apr 28, 2000 |
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08962523 |
Oct 31, 1997 |
5997862 |
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09395636 |
Sep 14, 1999 |
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Current U.S.
Class: |
424/94.2 ;
424/94.1 |
Current CPC
Class: |
Y02A 50/481 20180101;
A61K 38/47 20130101; A61K 38/162 20130101; Y02A 50/30 20180101;
Y02A 50/473 20180101; A61K 38/162 20130101; A61K 2300/00 20130101;
A61K 38/47 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/094.2 ;
424/094.1 |
International
Class: |
A61K 38/43 20060101
A61K038/43; A61K 38/54 20060101 A61K038/54 |
Claims
1) A method for the prophylactic or therapeutic treatment of
bacterial infections, comprising: administering to the site of the
infection an effective amount of a modified version of at least one
lytic enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof
2) The method according to claim 1, further comprising
administering at least one holin enzyme with said at least one
lytic enzyme.
3) The method according to claim 2, wherein said at least one holin
enzyme is a shuffled holin enzyme.
4) The method according to claim 2, wherein said holin enzyme is a
chimeric holin enzyme.
5) The method according to claim 1, further comprising at least one
lytic enzyme which is neither said shuffled lytic enzyme nor said
chimeric lytic enzyme.
6) The method according to claim 1, further comprising delivering
said modified lytic enzyme in a carrier suitable for delivering
said lytic enzyme to the site of the infection.
7) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Hemophilus
influenza.
8) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Pseudomonas.
9) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Streptococcus
pneumoniae
10) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of
Streptococcusfasciae
11) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Listeria.
12) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Salmonella.
13) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of E. coli.
14) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Campylobacter.
15) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Pseudomonas.
16) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Streptococcus
mutans.
17) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Mycobacterium
tuberculosis.
18) The method according to claim 1, wherein the at least one
modified lytic enzyme is for the treatment of Streptococcus.
19) The method according to claim 6, wherein the carrier is an
inhalant.
20) The method according to claim 6, wherein the carrier is a
topical cream
21) The method according to claim 6, wherein the carrier is a nasal
spray.
22) The method according to claim 6, wherein the carrier is a
syrup.
23) The method according to claim 6, wherein the carrier is a
tablet.
24) The method according to claim 6, wherein the carrier is a
tampon.
25) The method according to claim 6, wherein the carrier is a
suppository.
26) The method according to claim 6, wherein the carrier is an eye
drop solution.
27) The method according to claim 6, wherein the carrier is a
candy.
28) The method according to claim 6, wherein the carrier is a
chewing gum.
29) The method according to claim 6, wherein the carrier is a
lozenge.
30) The method according to claim 6, wherein the carrier is a
troche.
31) The method according to claim 6, wherein the carrier is a
powder.
32) The method according to claim 6, wherein the carrier is an
aerosol.
33) The method according to claim 6, wherein the carrier is a
liquid.
34) The method according to claim 6, wherein the carrier is a
liquid spray.
35) The method according to claim 6, wherein the carrier is a
bandage.
36) The method according to claim 6, wherein the carrier is a
toothpaste.
37) The method according to claim 6, wherein the carrier is an oral
wash.
38) A method for the prophylactic and therapeutic treatment of
bacterial infections of an upper respiratory tract, comprising
administering a composition comprising an effective amount of a
modified version of a lytic enzyme produced by a bacteria infected
with a bacteriophage specific for said bacteria wherein said
modified version of said at least one lytic enzyme is selected from
the group consisting of shuffled lytic enzymes, chimeric lytic
enzymes, and combinations thereof.
39) The method according to claim 38, further comprising
administering a holin enzyme with said modified lytic enzyme.
40) The method according to claim 39, wherein said holin enzyme is
a shuffled holin enzyme.
41) The method according to claim 39, wherein said holin enzyme is
a chimeric holin enzyme.
42) The method according to claim 38, further comprising at least
one lytic enzyme which is neither said shuffled lytic enzyme nor
said chimeric lytic enzyme.
43) The method according to claim 38, further comprising delivering
said modified lytic enzyme in a carrier suitable for delivering
said lytic enzyme to the mouth, the throat or the nasal
passage.
44) The method according to claim 38, wherein said bacteria being
treated is Streptococcus pneumoniae.
45) The method according to claim 38, wherein said bacteria being
treated is Hemophilus influenza.
46) The method according to claim 43, wherein said carrier is a
candy, chewing gum, lozenge, troche, tablet, a powder, an aerosol,
a liquid and a liquid spray.
47) The method according to claim 38, wherein said composition
further comprises a buffer that maintains pH of the composition at
a range between about 4.0 and about 9.0.
48) The method according to claim 48, wherein the buffer maintains
the pH of the composition at the range between about 5.5 and about
7.5.
49) The method according to claim 47, wherein said buffer comprises
a reducing reagent.
50) The method according to claim 49, wherein said reducing reagent
is dithiothreitol.
51) The method according to claim 47, wherein said buffer comprises
a metal chelating reagent.
52) The method according to claim 51, wherein said metal chelating
reagent is ethylenediaminetetracetic disodium salt.
53) The method according to claim 47, wherein said buffer is a
citrate-phosphate buffer.
54) The method according to claim 38, wherein said at least one
modified lytic enzyme is lyophilized.
55) The method according to claim 43, wherein said carrier further
comprises a sweetener.
56) The method according claim 38, further comprising administering
a concentration of about 100 to about 100,000 active enzyme units
per milliliter of fluid in the wet environment of the nasal or oral
passages.
57) The method according to claim 56, further comprising
administering the concentration of about 100 to about 10,000 active
enzyme units per milliliter of fluid in the wet environment of the
nasal or oral passages.
58) The method according to claim 38, further comprising using said
composition in the therapeutic treatment of Streptococcus
infections.
59) The method according to claim 38, further comprising using said
composition in the prophylactic treatment of Streptococcus
infections.
60) The method according to claim 38, further comprising using said
composition in the prophylactic treatment of Hemophilus
infections.
61) The method according to claim 38, further comprising using said
composition in the therapeutic treatment of Hemophilus
infections.
62) A composition for use in the therapeutic or prophylactic
treatment of a bacterial infection of an upper respiratory tract,
comprising: an effective amount of a modified version of a lytic
enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof, and a carrier for delivering said at least one lytic
enzyme to a mouth, throat, or nasal passage.
63) The composition according to claim 62, further comprising a
holin enzyme.
64) The composition according to claim 63, wherein said holin
enzyme is a shuffled holin enzyme.
65) The composition according to claim 63, wherein said holin
enzyme is a chimeric holin enzyme.
66) The composition according to claim 62, wherein said bacteria
being treated is Streptococcus pneumoniae.
67) The composition according to claim 62, wherein said bacteria
being treated is Hemophilus influenza.
68) The composition according to claim 62, wherein said carrier is
selected from the group consisting of a candy, chewing gum,
lozenge, troche, tablet, a powder, an aerosol, a liquid and a
liquid spray.
69) The composition according to claim 62, wherein said composition
further comprises a buffer that maintains pH of the composition at
a range between about 4.0 and about 9.0.
70) The composition according to claim 62, wherein the buffer
maintains the pH of the composition at the range between 5.5 and
7.5.
71) The composition according to claim 62, further comprising a
bactericidal or bacteriostatic agent 5 as a preservative.
72) The composition according to claim 62, wherein said modified
lytic enzyme is lyophilized.
73). The composition according claim 62, wherein said at least one
lytic enzyme is present in a concentration of about 100 to about
100,000 active enzyme units per milliliter of fluid in the wet
environment of the nasal or oral passages.
74). The composition according to claim 71, wherein said at least
one lytic enzyme is present in a concentration of about 100 to
about 10,000 active enzyme units per milliliter of fluid in the wet
environment of the nasal or oral passages.
75) A method for the treatment of bacterial infections of the
digestive tract, comprising administering to the digestive tract a
composition comprising an effective amount of a modified version of
a lytic enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof.
76) The method according to claim 75, further comprising
administering a holin enzyme with said modified lytic enzyme.
77) The method according to claim 76, wherein said holin enzyme is
a shuffled holin enzyme. 5
78) The method according to claim 76, wherein said holin enzyme is
a chimeric holin enzyme.
79) The method according to claim 75, further comprising delivering
said lytic enzyme in a carrier suitable for delivering said lytic
enzyme to the digestive tract. 10
80) The method according to claim 75, wherein said bacterial
infections are caused by gram negative bacteria selected from the
group consisting of Listeria, Salmonella, E. coli, and
Campylobacter.
81) The method according to claim 73, wherein said carrier is
selected from the group consisting of suppository enemas, syrups,
and enteric coated pills.
82) A composition for treating for the treatment of bacterial
infections of the digestive tract, comprising: an effective amount
of a modified version of a lytic enzyme produced by a bacteria 20
infected with a bacteriophage specific for said bacteria wherein
said modified version of said at least one lytic enzyme is selected
from the group consisting of shuffled lytic enzymes, chimeric lytic
enzymes, and combinations thereof, and a carrier for delivering
said lytic enzyme to the digestive tract.
83) The composition according to claim 82, further comprising a
holin enzyme
84) The composition according to claim 83, wherein said holin
enzyme is a shuffled holin enzyme.
85) The composition according to clai 84, wherein said holin enzyme
is a chimeric holin enzyme.
86) The composition according to claim 82, wherein said bacteria to
be treated are selected from the group consisting of Listeria,
Salmonella, E. coli, and Campylobacter.
87) The composition according to claim 82, wherein said carrier for
delivering said at least one modified lytic enzyme to the digestive
tract is selected from the group consisting of suppository enemas,
syrups, or enteric coated pills.
88) The composition according to claim 82, wherein said composition
further comprises a buffer that maintains pH of the composition at
a range between about 4.0 and 9.0.
89). The composition according claim 82, wherein said at least one
modifed lytic enzyme is present in a concentration of about 100 to
about 100,000 active enzyme units per milliliter of fluid in the
wet environment of the digestive tract
90) The composition according to claim 89, wherein said at least
one modified lytic enzyme is present in a concentration of about
100 to about 10,000 active enzyme units per milliliter of fluid in
the wet environment of the digestive tract.
91) A composition for the therapeutic or prophylactic treatment of
bacterial infections of bums and wounds of the skin, comprising: an
effective amount of a modified version of a lytic enzyme produced
by a bacteria infected with a bacteriophage specific for said
bacteria wherein said modified version of said at least one lytic
enzyme is selected from the group consisting of shuffled lytic
enzymes, chimeric lytic enzymes, and combinations thereof; and a
carrier for delivering said at least one lytic enzyme to the
skin.
92) The composition according to claim 91, further comprising a
holin enzyme.
93) The composition according to claim 92, wherein said holin
enzyme is a shuffled holin enzyme.
94) The composition according to claim 92, wherein said holin
enzyme is a chimeric holin enzyme.
95) The composition according to claim 91, wherein said carrier is
a bandage.
96) The composition according to claim 91, further comprising using
said composition in the prophylactic treatment of bacterial
infections.
97) The composition according to claim 91, further comprising using
said composition in the therapeutic treatment of bacterial
infections.
98) The composition according to claim 91, wherein said bacteria
being treated is Pseudomonas.
99) The composition according to claim 91, wherein said bacteria
being treated is Staphylococcus.
100) The composition according to claim 91, wherein said bacterium
being treated are Staphylococcus and Pseudomonas.
101) A method for the therapeutic or prophylactic treatment of
bacterial infections of bums and wounds of the skin, comprising:
administering to an infected area of the skin a composition
comprising an effective amount of a modified version of a lytic
enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof
102) The composition according to claim O1, further comprising a
holin enzyme.
103) The composition according to claim 102, wherein said holin
enzyme is a shuffled holin enzyme.
104) The composition according to claim 102, wherein said holin
enzyme is a chimeric holin enzyme.
105) The method according to claim 101, further comprising
delivering said at least one modified lytic enzyme in a carrier
suitable for delivering said lytic enzyme to the skin.
106) The method according to claim 105, wherein said carrier is a
bandage.
107) The method according to claim 101, further comprising using
said composition in the prophylactic treatment of bacterial
infections.
108) The method according to claim 101, further comprising using
said composition in the therapeutic treatment of bacterial
infections.
109) The method according to claim 101, wherein said bacteria being
treated is Pseudomonas.
110) The method according to claim 101, wherein said bacteria being
treated is Staphylococcus.
111) The method according to claim 101, wherein said bacterium
being treated are Staphylococcus and Pseudomonas.
112) A method for the prophylactic and therapeutic treatment of
vaginal infections, comprising: administering to the vagina
composition an effective amount of a modified version of a lytic
enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof.
113) The method according to claim 1 12, further comprising
administering a holin enzyme with said modified lytic enzyme.
114) The method according to claim 113, wherein said holin enzyme
is a shuffled holin enzyme.
115) The method according to claim 113, wherein said holin enzyme
is a chimeric holin enzyme.
116) The method according to claim 11 1, further comprising
delivering said modified lytic enzyme in a carrier suitable for
delivering said modified lytic enzyme to the vagina.
117) The method according to claim 115, wherein said carrier is to
be placed in the vagina.
118) The method according to claim 116, wherein said carrier is a
tampon.
119) The method according to claim 1 16, wherein said carrier is a
pad.
120) The method according to claim 116, wherein said carrier is a
douche.
121) The method according to claim 112, wherein said lytic enzyme
is specific for Group B Streptococcus.
122) A composition for the prophylactic and therapeutic treatment
of treatment of vaginal infections, comprising: an effective amount
of a modified version of a lytic enzyme produced by a bacteria
infected with a bacteriophage specific for said bacteria wherein
said modified version of said at least one lytic enzyme is selected
from the group consisting of shuffled lytic enzymes, chimeric lytic
enzymes, and combinations thereof; and a carrier for delivering
said lytic enzyme to a vagina.
123) The method according to claim 122, further comprising
administering a holin enzyme with said modified lytic enzyme.
124) The method according to claim 123, wherein said holin enzyme
is a shuffled holin enzyme.
125) The method according to claim 122, wherein said holin enzyme
is a chimeric holin enzyme.
126) The composition according to claim 122, wherein said carrier
is a tampon.
127) The composition according to claim 122, wherein said carrier
is a douche.
128) The composition according to claim 122, wherein said carrier
is a pad.
129) The composition according to claim 122, wherein said lytic
enzyme is specific for Group B Streptococcus.
130) A method for the prophylactic and therapeutic treatment of eye
infections, comprising: administering to an eye a composition
comprising an effective amount of a modified version of a lytic
enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof.
131) The method according to claim 130, further comprising
administering a holin enzyme with said modified lytic enzyme.
132) The method according to claim 131, wherein said holin enzyme
is a shuffled holin enzyme.
133) The method according to claim 131, wherein said holin enzyme
is a chimeric holin enzyme.
134) The method according to claim 130, further comprising
delivering said lytic enzyme in a carrier suitable for delivering
said lytic enzyme to the eye.
135) The method according to claim 130, wherein said bacteria being
treated is Hemophilus.
136) The method according to claim 130, wherein said bacteria being
treated is Staphylococcus.
137) The method according to claim 134, wherein the carrier is an
eye drop solution.
138) The method according to claim 134, wherein the carrier is an
eye wash solution.
139) The method according to claim 138, wherein said solution is an
isotonic solution.
140) A composition for use in the therapeutic or prophylactic
treatment of an eye infection,, comprising an effective amount of a
modified version of a lytic enzyme produced by a bacteria infected
with a bacteriophage specific for said bacteria wherein said
modified version of said at least one lytic enzyme is selected from
the group consisting of shuffled lytic enzymes, chimeric lytic
enzymes, and combinations thereof; and a carrier for delivering
said lytic enzyme to the eye.
141) The composition according to claim 140, further comprising
administering a holin enzyme with said modified lytic enzyme.
142) The composition according to claim 141, wherein said holin
enzyme is a shuffled holin enzyme.
143) The composition according to claim 141, wherein said holin
enzyme is a chimeric holin enzyme.
144) The composition according to claim 140, wherein said bacteria
being treated is Hemophilus.
145) The composition according to claim 140, wherein said bacteria
being treated is Staphylococcus.
146) The composition according to claim 140, wherein said carrier
is an isotonic solution.
147) The composition according to claim 146, wherein said isotonic
solution is in an eye drop dispenser.
148) A method for the prophylactic or therapeutic treatment of
dermatological infections comprising: topically applying to an
infected area of the skin a composition comprising an effective
amount of a modified version of a lytic enzyme produced by a
bacteria infected with a bacteriophage specific for said bacteria
wherein said modified version of said at least one lytic enzyme is
selected from the group consisting of shuffled lytic enzymes,
chimeric lytic enzymes, and combinations thereof.
149) The method according to claim 148, further comprising
administering a holin enzyme with said modified lytic enzyme.
150) The method according to claim 149, wherein said holin enzyme
is a shuffled holin enzyme.
151) The method according to claim 149, wherein said holin enzyme
is a chimeric holin enzyme.
152) The method according to claim 148, further comprising
delivering said composition in a pharmaceutically acceptable
carrier.
153) The method according to claim 152, wherein said carrier is
selected from the group consisting of an aqueous liquid, an alcohol
base, a water soluble gel, a lotion, an ointment, a nonaqueous
liquid base, a mineral oil base, a blend of mineral oil and
petrolatum, lanolin, liposomes, hydrophilic gelling agents,
cross-linked acrylic acid polymers (carbomer), cellulose polymers,
hydroxy ethyl cellulose, cellulose gum, MVE/MA decadiene
crosspolymers, PVM/MA copolymers, and any combinations thereof.
154) The method according to claim 152, wherein the form in which
the composition is delivered is selected from the group consisting
of a spray, a smear, a time release patch, a liquid absorbed wipe,
and any combinations thereof.
155) The method according to claim 148, wherein the lytic enzyme is
in an environment having a pH which allows for activity of said
lysin enzyme.
156) The method according to claim 155, wherein said composition
further comprises a buffer that maintains pH of the composition at
a range between about 4.0 and about 9.0.
157) The method according to claim 148, wherein said composition
further comprises a mild surfactant in an amount effective to
potentiate effects of the lytic enzyme.
158) The method according to claim 148, wherein the composition
further comprises at least one complementary agent which
potentiates the bactericidal activity of the lytic enzyme, said
complementary agent being selected from the group consisting of
penicillin, synthetic penicillins bacitracin, methicillin,
cephalosporin, polymyxin, cefaclor. Cefadroxil, cefamandole nafate,
cefazolin, cefixime, cefmetazole, cefonioid, cefoperazone,
ceforanide, cefotanme, cefotaxime, cefotetan, cefoxitin,
cefpodoxime proxetil, ceftazidime, ceftizoxime, ceftriaxone,
cefriaxone moxalactam, cefuroxime, cephalexin, cephalosporin C,
cephalosporin C sodium salt, cephalothin, cephalothin sodium salt,
cephapirin, cephradine, cefuroximeaxetil, dihydratecephalothin,
moxalactam, loracarbef. mafate and chelating agents in an amount
effective to synergistically enhance effects of the lytic
enzyme.
159) The method according to claim 148, wherein the composition
further comprises lysostaphin for the treatment of any
Staphylococcus aureus bacteria.
160) The method according to claim 148, wherein said lytic enzyme
is present in an amount ranging from about 100 to about 500,000
units per milliliter.
161) A composition for the treatment of dermatological infections
comprising: an effective amount of a modified version of a lytic
enzyme produced by a bacteria infected with a bacteriophage
specific for said bacteria wherein said modified version of said at
least one lytic enzyme is selected from the group consisting of
shuffled lytic enzymes, chimeric lytic enzymes, and combinations
thereof; and a carrier.
162) The composition according to claim 161, wherein said carrier
is selected from the group consisting of an aqueous liquid, an
alcohol base, a water soluble gel, a lotion, an ointment, a
nonaqueous liquid base, a mineral oil base, a blend of mineral oil
and petrolatum, lanolin, liposomes, hydrophilic gelling agents,
cross-linked acrylic acid polymers (carbomer), cellulose polymers,
hydroxy ethyl cellulose, cellulose gum, MVE/MA decadiene
crosspolymers, PVM/MA copolymers, and any combinations thereof.
163) The composition according to claim 161, wherein said
composition is in the form selected from the group consisting of a
spray, a smear, a time release patch, a liquid absorbed wipe, and
any combinations thereof.
164) The composition according to claim 161, wherein the at least
one lytic enzyme is in an environment having a pH which allows for
activity of said lytic enzyme.
165) The composition according to claim 164, wherein said
composition further comprises a buffer 5 that maintains pH of the
composition at a range between about 4.0 and about 9.0.
166) The composition according to claim 165, wherein said buffer
maintains the pH of the composition at the range of between about
5.5 and about 7.5.
167) The composition according to claim 166, further comprising a
bactericidal or bacteriostatic agent as a preservative.
168) The composition according to claim 167, further comprising a
surfactant in an amount effective to potentiate a therapeutic
effect of the composition.
169) The composition according to claim 161, wherein the
composition further comprises at least one complementary agent
which potentiates the bactericidal activity of the lytic enzyme,
said complementary agent being selected from the group consisting
of penicillin, synthetic penicillins bacitracin, methicillin,
cephalosporin, polymyxin, cefaclor. Cefadroxil, cefamandole nafate,
cefazolin, cefixime, cefmetazole, cefonioid, cefoperazone,
ceforanide, cefotanme, cefotaxime, cefotetan, cefoxitin,
cefpodoxime proxetil, ceftazidime, ceftizoxime, ceftriaxone,
cefriaxone moxalactam, cefuroxime, cephalexin, cephalosporin C,
cephalosporin C sodium salt, cephalothin, cephalothin sodium salt,
cephapirin, cephradine, cefuroximeaxetil, dihydratecephalothin,
moxalactam, loracarbef. mafate chelating agents, and combinations
thereof in an amount effective to synergistically enhance the
therapeutic effect of the lytic enzyme.
170) The composition according to claim 161, wherein the
composition further comprises lysostaphin for the treatment of any
Staphylococcus aureus bacteria.
171) The composition according to claim 161, wherein the
composition further comprises lysozyme.
172) The composition according to claim 161, further comprising at
least one emulsifier.
173) The composition according to claim 161, further comprising at
least one antioxidant.
174) The composition according to claim 161, further comprising at
least one sunscreen.
175) The composition according to claim 161, further comprising at
least one anti-inflammatory agent.
176) A composition for the therapeutic or prophylactic treatment of
bacterial infections of the upper respiratory system, comprising an
effective amount of a modified version of a lytic enzyme produced
by a bacteria infected with a bacteriophage specific for said
bacteria wherein said modified version of said at least one lytic
enzyme is selected from the group consisting of shuffled lytic
enzymes, chimeric lytic enzymes, and combinations thereof, and a
pharmaceutically acceptable carrier in an inhaler allowing for the
administration of the at least one lytic enzyme to the bronchial
tubes and lungs.
177) The composition according to claim 176, further comprising
administering a holin enzyme with said modified lytic enzyme.
178) The composition according to claim 177, wherein said holin
enzyme is a shuffled holin enzyme.
179) The composition according to claim 177, wherein said holin
enzyme is a chimeric holin enzyme.
180) The composition according to claim 176, wherein said
composition is for the therapeutic treatment of bacterial
infections of the upper respiratory system.
181) The composition according to claim 176, wherein said
composition is for the prophylactic treatment of bacterial
infections of the upper respiratory system.
182) A composition for the therapeutic or prophylactic treatment of
bacterial infections of the mouth or teeth, comprising an effective
amount of a modified version of a lytic enzyme produced by a
bacteria infected with a bacteriophage specific for said bacteria
wherein said modified version of said at least one lytic enzyme is
selected from the group consisting of shuffled lytic enzymes,
chimeric lytic enzymes, and combinations thereof and a
pharmaceutically acceptable carrier for topical application of the
at least one lytic enzyme.
183) The composition according to claim 182, further comprising
administering a holin enzyme with said modified lytic enzyme.
184) The method according to claim 183, wherein said holin enzyme
is a shuffled holin enzyme.
185) The method according to claim 183, wherein said holin enzyme
is a chimeric holin enzyme.
186) The composition according to claim 182, wherein said
composition is used for the prophylactic treatment of dental
caries.
187) The composition according to claim 182, wherein said
composition is used for the therapeutic treatment of dental
caries.
188) The composition according to claim 182, wherein said carrier
is toothpaste.
189) The composition according to claim 182, wherein said carrier
is an oral wash.
190) The composition according to claim 182, wherein said carrier
is a chewing gum.
191) The composition according to claim 182, wherein said carrier
is a lozenge.
192) The composition according to claim 182, wherein said bacteria
being treated is Streptococcus mutans.
193) The composition according to claim 182, wherein said lytic
enzyme is present in an amount ranging from about 100 to about
500,000 units per milliliter.
194) The composition according to claim 193, wherein said lytic
enzyme is present in an amount ranging from about 10,000 to about
100,000 units per milliliter.
Description
[0001] The following application is a continuation-in-part of U.S.
patent application Ser. No. 09/395,636 filed Sep. 14, 1999, which
is a continuation of continuation of U.S. patent application Ser.
No. 08/962,523, filed Oct. 31, 1997, now U.S. Pat. No.
5,997,862.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention discloses a method and composition for
the treatment of bacterial infections by the use of a lytic enzymes
and holin enzymes blended with an appropriate carrier suitable for
the treatment of the infection.
[0004] 2. Description of the Prior Art
[0005] In the past, antibiotics have been used to treat various
infections. The work of Selman Waksman in the introduction and
production of Streptomycetes, Dr. Fleming's discovery of
penicillin, are well known as well as the work of numerous others
in the field of antibiotics. Over the years, there have been
additions and chemical modifications to the "basic" antibiotics in
attempts to make them more powerful, or to treat people allergic to
these antibiotics.
[0006] Others have found new uses for these antibiotics. U.S. Pat.
No. 5,260,292 (Robinson et al.) discloses the topical treatment of
acne with aminopenicillins. The method and composition for
topically treating acne and acneiform dermal disorders includes
applying an amount of an antibiotic selected from the group
consisting of ampicillin, amoxicillin, other aminopenicillins, and
cephalosporins, and derivatives and analogs thereof, effective to
treat the acne and acneiform dermal disorders. U.S. Pat. No.
5,409,917 (Robinson et al.) discloses the topical treatment of acne
with cephalosporins.
[0007] However, as more antibiotics have been prescribed or used at
an ever increasing rate for a variety of illnesses, increasing
numbers of bacteria have developed a resistance to antibiotics.
Larger doses of stronger antibiotics are now being used to treat
ever more resistant strains of bacteria. Multiple antibiotic
resistant bacteria have consequently developed. The use of more
antibiotics and the number of bacteria showing resistance has led
to increasing the amount of time that the antibiotics need to be
used. Broad, non-specific antibiotics, some of which have
detrimental effects on the patient, are now being used more
frequently. Also, antibiotics do not easily penetrate mucus
linings.
Additionally, the number of people allergic to antibiotics appears
to be increasing.
[0008] Consequently, other efforts have been sought to first
identify and then kill bacteria.
[0009] Attempts have been made to treat bacterial diseases with by
the use of bacteriophages. U.S. Pat. No. 5,688,501 (Merril, et al.)
discloses a method for treating an infectious disease caused by
bacteria in an animal with lytic or non-lytic bacteriophages that
are specific for particular bacteria.
[0010] U.S. Pat. No. 4,957,686 (Norris) discloses a procedure of
improved dental hygiene which comprises introducing into the mouth
bacteriophages parasitic to bacteria which possess the property of
readily adhering to the salivary pellicle.
[0011] It is to be noted that the direct introduction of
bacteriophages into an animal to prevent or fight diseases has
certain drawbacks. Specifically, the bacteria must be in the right
growth phase for the phage to attach. Both the bacteria and the
phage have to be in the correct and synchronized growth cycles.
Additionally, there must be the right number of phages to attach to
the bacteria; if there are too many or too few phages, there will
either be lysis from without or no production of the lysing enzyme,
respectively. The phage must also be active where damaged phage are
inactive. The phages are also inhibited by many things including
bacterial debris from the organism it is going to attack. Further
complicating the direct use of bacteriophage to treat bacterial
infections is the possibility of immunological reactions, rendering
the phage non-functional.
[0012] Consequently, others have explored the use of other safer
and more effective means to treat and prevent bacterial
infections.
[0013] U.S. Pat. No. (application Ser. No. 08/962,523) (Fischetti,
et. al.) and U.S. Pat. No. (application Ser. No. 09/257,026)
(Fischetti et al.) disclose the use of an oral delivery mode, such
as a candy, chewing gum, lozenge, troche, tablet, a powder, an
aerosol, a liquid or a liquid spray, containing a lysin enzyme
produced by group C streptococcal bacteria infected with a C1
bacteriophage for the prophylactic and therapeutic treatment of
Streptococcal A throat infections, commonly known as strep
throat.
[0014] U.S. patent Ser. No. 09/395,636 (Fischetti et al.) discloses
a method and composition for the prophylactic or therapeutic
treatment of bacterial infections, comprising administering an
effective amount of at least one lytic enzyme produced by a
bacteria infected with a bacteriophage specific for the bacteria to
the site of the infection. The lytic enzyme preferably comprises a
carrier suitable for delivering the lytic enzyme to the site of the
infection. This method and treatment may be used for treating upper
respiratory infections, topical infections, vaginal infections, eye
infections ear infections, for parenteral treatment, and for most
other bacterial infections.
SUMMARY OF THE INVENTION
[0015] The method for obtaining and purifying the lytic enzyme
produced by a bacteria infected with the bacteriophage is known in
the art. While many lytic enzymes are phage encoded, some recent
evidence suggests that phage enzymes may actually be a bacterial
enzyme that is used to construct the cell wall and the phage. While
replicating in the bacterium, a phage gene product may cause the
upregulation or derepression of bacterial enzyme for the purpose of
releasing the bacteriophage. These bacterial enzymes may be tightly
regulated by the bacterial cell and are used by the bacteria for
the construction and assembly of the cell wall.
[0016] The use of these lytic enzymes for the prophylactic and
therapeutic treatment of bacterial diseases, however, has not been
explored, except by the inventors of the present invention.
Consequently, the present invention discloses the extraction and
use of a variety of bacterial phage associated lytic enzymes, holin
enzymes, chimeric enzymes, and shuffled enzymes for the treatment
of a wide variety of illnesses caused by bacterial infections.
[0017] The use of phage associated lytic enzymes produced by the
infection of a bacteria with a bacteria specific phage has numerous
advantages for the treatment of diseases. As the phage are targeted
for specific bacteria, the lytic enzymes do not interfere with
normal flora. The actions of the lytic enzymes are fast and do not
depend on bacterial growth.
[0018] Lytic enzymes can be directed to the mucosal lining, where,
in residence, they will be able to kill colonizing disease
bacteria.
[0019] Shuffled enzymes are enzymes in which the gene for more than
one related lytic enzymes have been randomly cleaved and
reassembled into a more active or specific enzyme.
[0020] In a preferred embodiment of the invention, shuffled enzymes
are used to treat bacterial infections, thereby increasing the
speed and efficiency with which the the bacteria are killed.
[0021] Chimeric enzymes are enzymes which are a combination of two
or more enzymes having two or more active sites such that the
chimeric enzyme can act independently on the same or different
molecules. This will allow for potentially treating two or more
different bacterial infections at the same time. Chimeric enzymes
may also be used to treat one bacterial infection by cleaving the
cell wall in more than one location.
[0022] A number of chimeric lytic enzymes have been produced and
studied. Gene E-L, a chimeric lysis constructed from bacteriophages
phi X174 and MS2 lysis proteins E and L, respectively, was
subjected to internal deletions to create a series of new E-L
clones with altered lysis or killing properties. The lytic
activities of the parental genes E, L, E-L, and the internal
truncated forms of E-L were investigated in this study to
characterize the different lysis mechanism, based on differences in
the architecture of the different membranes spanning domains.
Electron microscopy and and release of marker enzymes for the
cytoplasmic and periplasmic spaces revealed that two different
lysis mechanisms can be distinguished depending on penetrating of
the proteins of either the inner membrane or the inner and outer
membranes of the E. coli. FEMS Microbiol. Lett. Jul. 1, 1998,
164(1); 159-67.
[0023] In another experiment an active chimeric cell wall lytic
enzyme (TSL) has been constructed by fusing the region coding for
the N-terminal half of the lactococcal phage Tuc2009 lysin and the
region coding for the C-terminal domain of the major pneumococcal
autolysin. The chimeric enzyme exhibited a glycosidase activity
capable of hydrolysing choline-containing pneumoccal cell
walls.
[0024] A preferred embodiment of this invention discloses the use
of chimeric lytic enzymes to treat two infectious bacteria at the
same time, or to cleave the cell wall of a bacteria in two
different locations.
[0025] Holin enzymes produce holes in the cell membrane. More
specifically, holins form lethal membrane lesions that terminates
respiration. Like the lytic enzymes, the holin enzymes are coded
for and carried by a phage. In fact, it is quite common for the
genetic code for the holin enzyme to be found next to or even
within the code for the lytic enzyme in the phage. Most holin
sequences are short, and overall, hydrophobic in nature, with a
highly hydrophilic carboxy-terminal domain. In many cases, the
putative holin is encoded on a different reading frame within the
enzymatically active domain of the phage. In other cases, the holin
is encoded on the DNA next or close to the DNA coding for the cell
wall lytic enzyme. The holin is frequently synthesized during the
late stage of phage infection and found in the cytoplasmic membrane
where it causes membrane lesions.
[0026] Holins can be grouped into two general classes based on
primary structure analysis. Class I holins are usually 95 residues
or longer and may have three potential transmembrane domains. Class
II holins are usually smaller, at approximately 65-95 residues, and
the distribution of charged and hydrophobic residues indicating two
TM domains (Young, et al. Trends in Microbiology v. 8, No. 4, March
2000). At least for the phages of gram-positive hosts, however, the
dual-component lysis system may not be universal. Although the
presence of holins has been shown or suggested for several phages,
no genes have yet been found encoding putative holins for all of
the phages. Holins have been shown to be present or suggested for
among others, lactococcal bacteriophage Tuc2009, lactococcal
.phi.LC3, pneumococcal bacteriophage EJ-1, Lactobacillus gasseri
bacteriophage .phi.adh, Staphylococcus aureus bacteriophage Twort,
Listeria monocytogenes bacteriophages, pneumococcal phage Cp-1,
Bacillus subtillis phage .phi.29, Lactobacillus delbrueckki
bacteriophage LL-H lysin, and bacteriophage .phi.11 of
Staphylococcus aureus. (Loessner, et al., Journal of Bacteriology,
August 1999, p. 4452-4460).
[0027] In another embodiment of the invention, holin enzymes are
used in conjunction with the lytic enzymes to accelerate the speed
and efficiency at which the bacteria are killed. Holin enzymes may
also be in the form of chimeric and/or shuffled enzymes. Holin
enzymes may also be used alone in the treatment of bacterial
infections
[0028] It is an object of the invention to use phage associated
lytic enzymes in combination with chimeric or shuffled lytic
enzymes to prophylactically and therapeutically treat bacterial
diseases.
[0029] In another embodiment of the invention, chimeric lytic
enzymes are used to prophylactically and therapeutically treat
bacterial diseases.
[0030] In yet another embodiment of the invention, shuffled lytic
enzymes are used to prophylactically and therapeutically treat
bacterial infections.
[0031] In yet another embodiment of the invention, holin enzymes
are used in conjunction with phage associated lytic enzymes to
prophylactically and therapeutically treat bacterial
infections.
[0032] In another embodiment of the invention, holin enzymes alone
are used to prophylactically and therapeutically treat bacterial
infections.
[0033] In another embodiment of the invention, the holin enzymes
are shuffled holin enzymes or chimeric holin enzymes, either in
combination with or independent of the lytic enzymes.
[0034] The invention (which incorporates U.S. Pat. No. 5,604,109 in
its entirety by reference) uses a lytic enzyme produced by the
bacterial organism after being infected with a particular
bacteriophage as either a prophylactic treatment for preventing
those who have been exposed to others who have the symptoms of an
infection from getting sick, or as a therapeutic treatment for
those who have already become ill from the infection. The present
invention is based upon the discovery that phage lytic enzymes
specific for bacteria infected with a specific phage can
effectively and efficiently break down the cell wall of the
bacterium in question. At the same time, the semipurified enzyme is
lacking in proteolytic enzymatic activity and therefore
non-destructive to mammalian proteins and tissues when present
during the digestion of the bacterial cell wall. As discussed
above, the lytic enzymes may be chimeric, shuffled or "natural,"
and may be in combination with at least one holin enzyme, which may
also be chimeric, shuffled, or "natural."
[0035] In one embodiment of the invention, the prophylactic and
therapeutic treatment of a variety of illnesses caused by
Streptococcal pyogenes, Streptococcal pneumoniae, Streptococcus
fasciae, and Hemophilus influenza are disclosed. In another
embodiment of the invention, gram negative bacterial infections
caused by Listeria, Salmonella, E. coli, and Campylobacter, are
treated by the use of lytic enzymes. These and other bacteria,
which can infect the digestive system, can be treated by
incorporating the lytic enzymes in suppository enemas, in syrups,
or in other carriers to get directly to the site of the
infection(s).
[0036] In another embodiment of the invention, lytic enzymes are
incorporated into bandages to prevent or treat infections of burns
and wounds. In yet another embodiment of the invention, the lytic
enzymes of phage associated with Staphylococcus or Pseudomonas are
incorporated into bandages to prevent or treat infections of burns
and wounds.
[0037] Vaginal infections caused by Group B Streptococcus can cause
premature birth and subsequent complications resulting in neonatal
sepsis. Lysin incorporated into tampons specific for group B strep
would prevent infection of the neonate during birth without
disturbing normal vaginal flora so that women would not be overcome
by yeast infection as a result of antibiotic therapy.
[0038] In another embodiment of the invention, eye drops containing
lytic enzymes of Hemophilus, Pseudomonas, and/or Staphylococcus can
be used to directly treat eye infections. Treatment with lytic
enzymes are faster and more expedient than with antibiotics.
[0039] In yet another embodiment of the invention the phage
associated lytic enzyme is put into a carrier which is placed in an
inhaler to treat or prevent the spread of diseases localized in the
mucus lining of the oral cavity and lungs. Specific lytic enzymes
for tuberculosis have been isolated and can be used.
[0040] In another embodiment of the invention the lytic enzyme is
administered in the form of a candy, chewing gum, lozenge, troche,
tablet, a powder, an aerosol, a liquid, a liquid spray, or
toothpaste for the prevention or treatment of bacterial infections
associated with upper respiratory tract illnesses.
[0041] In another embodiment of the invention, species specific
lytic enzymes can be used in the treatment of bacterial infections
associated with topical or dermatological infections, administered
in the form of a topical ointment or cream. In another embodiment
of the invention, the lytic enzyme would be administered in an
aqueous form. In yet another embodiment of the invention,
lysostaphin, the enzyme which lyses Staphylococcus aureus, can be
included in the therapeutic agent. In a further embodiment of the
invention, conventional antibiotics may be included in the
therapeutic agent with the lytic enzyme, and with or without the
presence of lysostaphin. More than one lytic enzyme may also be
included in the prophylactic or therapeutic agent.
DETAILED DESCRIPTION OF THE INVENTION
[0042] The method for treating bacterial infections comprises
treating the infection with a therapeutic agent comprising an
effective amount of a modified version of at least one lytic enzyme
produced by a bacteria infected with a bacteriophage specific for
the bacteria wherein the modified version of the one lytic enzyme
is selected from the group consisting of shuffled lytic enzymes,
chimeric lytic enzymes, and combinations thereof. The lytic enzyme
is preferably in an environment having a pH which allows for
activity of said lytic enzyme. A holin enzyme may be used in
conjunction with the administration of the modified lytic enzyme.
The holin enzyme may be in its "natural" state, may be shuffled
holin enzymes or may be chimeric lytic enzymes.
[0043] The shuffled and chimeric enzymes are usually produced
either enzymatically or through recombinant DNA means. Any method
may be used to produce these enzymes.
[0044] The lytic enzyme can be used for the treatment or prevention
of Streptococcal pygenes, Hemophilus influenza, Pseudomonas,
Streptococcus pneumoniae, Streptococcus fasciae, Streptococcus
group B, Listeria, Salmonella, E. coli, Campylobacter, and other
bacteria, and any combination thereof. This lytic enzyme may be
either supplemented by chimeric and/or shuffled lytic enzymes, or
may be itself a chimeric and/or shuffled lytic enzyme. Similarly, a
holin enzyme may be included, which may also be a chimeric and/or
shuffled lytic enzyme.
[0045] For example, if there is a bacterial infection of the upper
respiratory tract, the infection can be prophylactically or
therapeutically treated with a composition comprising an effective
amount of at least one lytic enzyme produced by a bacteria being
infected with a bacteriophage specific for that bacteria, and a
carrier for delivering the lytic enzyme to a mouth, throat, or
nasal passage. It is preferred that the lytic enzyme is in an
environment having a pH which allows for activity of the lytic
enzyme. If an individual has been exposed to someone with the upper
respiratory disorder, the lytic enzyme will reside in die mucosal
lining and prevent any colonization of the infecting bacteria.
[0046] Two examples of bacteria which infect the upper respiratory
system are Streptococcus pneumoniae and Hemophilus influenzae. In
recent years, there has been an increase in the number of people,
particularly children and the elderly, that are infected or are
carriers of penicillin resistant Streptococcus pneumoniae and
Hemophilus. While these bacteria are normally harmless residents of
the host, they are opportunistic organisms that are able to cause
infections when the resistance of the host has been compromised. By
eliminating or reducing the number of these organisms in the upper
respiratory tract, there will be a commensurate reduction in the
number of infections by these bacteria.
[0047] Infection of the Hemophilus bacteria by Bacteriophage HP1 (a
member of the P2-like phage family with strong similarities to
coliphages P2 and 186, and some similarity to the retronphage Ec67)
produces a lytic enzyme capable of lysing the bacteria. The lytic
enzyme for Streptococcus pneumoniae, previously identified as an
N-acetyl-muramoyl-L-alanine amidase, is produced by the infecting
Streptococcus pneumoniae with the Pal bacteriophage. The
therapeutic agent can contain either or both of the lytic enzymes
produced by these two bacteria, and may contain other lytic enzymes
for other bacteria. The composition which may be used for the
prophylactic and therapeutic treatment of a strep infection
includes the lysin enzyme and a means of application, (such as a
carrier system or an oral delivery mode), to the mucosal lining of
the oral and nasal cavity, such that the enzyme is put in the
carrier system or oral delivery mode to reach the mucosa lining.
Another infection which can be treated prophylactically
Streptococcus group A, which can produce what is commonly known as
"strep" throat. When group C Streptococci are infected with a C1
bacteriophage, a lysin enzyme is produced specific for the lysing
of Streptococcus group A.
[0048] Prior to, or at the time the lysin, enzyme is put in the
carrier system or oral delivery mode, it is preferred that the
enzyme be in a stabilizing buffer environment for maintaining a pH
range between about 4.0 and about 9.0, more preferably between
about 5.5 and about 7.5 and most preferably at about 6.1.
[0049] The stabilizing buffer should allow for the optimum activity
of the lysin enzyme. The buffer may be a reducing reagent, such as
dithiothreitol. The stabilizing buffer may also be or include a
metal chelating reagent, such as ethylenediaminetetracetic acid
disodium salt, or it may also contain a phosphate or
citrate-phosphate buffer.
[0050] Means of application include, but are not limited to direct,
indirect, carrier and special means or any combination of means.
Direct application of the lytic enzyme may be by nasal sprays,
nasal drops, nasal ointments, nasal washes, nasal injections, nasal
packings, bronchial sprays and inhalers, or indirectly through use
of throat lozenges, or through use of mouthwashes or gargles, or
through the use of ointments applied to the nasal nares, the bridge
of the nose, or the face or any combination of these and similar
methods of application. The forms in which the lysin enzyme may be
administered include but are not limited to lozenges, troches,
candies, injectants, chewing gums, tablets, powders, sprays,
liquids, ointments, and aerosols.
[0051] The lozenge, tablet, or gum into which the lytic enzyme is
added may contain sugar, corn syrup, a variety of dyes, non-sugar
sweeteners, flavorings, any binders, or combinations thereof.
Similarly, any gum based products may contain acacia, carnauba wax,
citric acid, corn starch, food colorings, flavorings, non-sugar
sweeteners, gelatin, glucose, glycerin, gum base, shellac, sodium
saccharin, sugar, water, white wax, cellulose, other binders, and
combinations thereof.
[0052] Lozenges may further contain sucrose, corn starch, acacia,
gum tragacanth, anethole, linseed, olcorcsin, mineral oil, and
cellulose, oiner binders, and combinations thereof. In another
embodiment of the invention, sugar substitutes are used in place of
dextrose, sucrose, or other sugars.
[0053] The enzyme may also be placed in a nasal spray, wherein the
nasal spray is the carrier. The nasal spray can be a long acting or
timed release spray, and can be manufactured by means well known in
the art. An inhalant may also be used, so that the phage enzyme may
reach further down into the bronchial tract, including into the
lungs.
[0054] Any of the carriers for the lytic enzyme may be manufactured
by conventional means. However, it is preferred that any mouthwash
or similar type products not contain alcohol to prevent denaturing
of the enzyme. Similarly, when the lytic enzyme is being placed in
a cough drop, gum, candy or lozenge during the manufacturing
process, such placement should be made prior to the hardening of
the lozenge or candy but after the cough drop or candy has cooled
somewhat, to avoid heat denaturation of the enzyme.
[0055] The enzyme may be added to these substances in a liquid form
or in a lyophilized state, whereupon it will be solubilized when it
meets body fluids such as saliva. The enzyme may also be in a
micelle or liposome.
[0056] The effective dosage rates or amounts of the lytic enzyme to
treat the infection will depend in part on whether the lytic will
be used therapeutically or prophylactically, the duration of
exposure of the recipient to the infectious bacteria, the size and
weight of the individual, etc. The duration for use of the
composition containing the enzyme also depends on whether the use
is for prophylactic purposes, wherein the use may be hourly, daily
or weekly, for a short time period, or whether the use will be for
therapeutic purposes wherein a more intensive regimen of the use of
the composition may be needed, such that usage may last for hours,
days or weeks, and/or on a daily basis, or at timed intervals
during the day. Any dosage form employed should provide for a
minimum number of units for a minimum amount of time. The
concentration of the active units of enzyme believed to provide for
an effective amount or dosage of enzyme may be in the range of
about 100 units/ml to about 100,000 units/ml of fluid in the wet or
damp environment of the nasal and oral passages, and possibly in
the range of about 100 units/ml to about 10,000 units/ml. More
specifically, time exposure to the active enzyme units may
influence the desired concentration of active enzyme units per ml.
It should be noted that carriers that are classified as "long" or
"slow" release carriers (such as, for example, certain nasal sprays
or lozenges) could possess or provide a lower concentration of
active (enzyme) units per ml, but over a longer period of time,
whereas a "short" or "fast" release carrier (such as, for example,
a gargle) could possess or provide a high concentration of active
(enzyme) units per ml, but over a shorter period of time. The
amount of active units per ml and the duration of time of exposure
depends on the nature of infection, whether treatment is to be
prophylactic or therapeutic, and other variables.
[0057] While this treatment may be used in any mammalian species,
the preferred use of this product is for a human.
[0058] This composition and method may also be used for the
treatment of Streptococcus A infections of the respiratory tract.
When using this composition for a Streptococcus A infection, the
lysin phage enzyme should be used for the prophylactic prevention
of Streptococcus infections. Similarly, in another embodiment of
the invention, this method may be used for the therapeutic and,
preferably, the prophylactic treatment of tuberculosis. In a
preferred embodiment of the invention, the phage associated lysing
enzyme for Mycobacteria tuberculosis is placed in a carrier in an
inhaler. The carrier may be sterile water or a water base, or any
other carrier used in an inhaler for dispersing drugs into the
bronchial tract. The phage associated lytic enzyme specific for
tuberculosis is subject to the same conditions as the phage
associated lytic enzyme for other lytic enzymes. Specifically,
prior to, or at the time the enzyme is put in the carrier system or
oral delivery mode, it is preferred that the enzyme be in a
stabilizing buffer environment for maintaining a pH range between
about 4.0 and about 9.0.
[0059] The stabilizing buffer should allow for the optimum activity
of the lytic enzyme. The buffer may be a reducing reagent, such as
dithiothreitol. The stabilizing buffer may also be or include a
metal chelating reagent, such as ethylenediaminetetracetic acid
disodium salt, or it may also contain a phosphate or
citrate-phosphate buffer.
[0060] For the prophylactic and therapeutic treatment of
tuberculosis, the phage associated lytic enzyme associated with
tuberculosis may also be applied by direct, indirect, carriers and
special means or any combination of means. Direct application of
the lytic enzyme may be by nasal sprays, nasal drops, nasal
ointments, nasal washes, nasal injections, nasal packings,
bronchial sprays and inhalers, or indirectly through use of throat
lozenges, or through use of mouthwashes or gargles, or through the
use of ointments applied to the nasal nares, the bridge of the
nose, or the face or any combination of these and similar methods
of application. The forms in which the lytic enzyme may be
administered include but are not limited to lozenges, troches,
candies, injectants, chewing gums, tablets, powders, sprays,
liquids, ointments, and aerosols. For the therapeutic treatment of
tuberculosis, the bronchial sprays and aerosols are most
beneficial, as these carriers, or means of distributing the
composition, allow the lytic enzyme to reach the bronchial tubes
and the lungs. An appropriate transport carrier may be attached to
the enzyme to transport the enzyme across the cell membrane to the
site of the bacteria.
[0061] The lozenge, tablet, or gum into which the lytic enzyme is
added may contain sugar, corn syrup, a variety of dyes, non-sugar
sweeteners, flavorings, any binders, or combinations thereof.
Similarly, any gum based products may contain acacia, carnauba wax,
citric acid, corn starch, food colorings, flavorings, non-sugar
sweeteners, gelatin, glucose, glycerin, gum base, shellac, sodium
saccharin, sugar, water, white wax, cellulose, other binders, and
combinations thereof.
[0062] Lozenges may further contain sucrose, corn starch, acacia,
gum tragacanth, anethole, linseed, oleoresin, mineral oil, and
cellulose, other binders, and combinations thereof. In another
embodiment of the invention, sugar substitutes are used in place of
dextrose, sucrose, or other sugars. However, to tackle bacterial
infections in the lung, the use of an inhaler carrier the lytic
enzyme in a carrier is preferred.
[0063] Another use of a lytic enzyme is for the treatment of
bacterial infections of the digestive tract. The method for
treating a bacterial infection of the digestive tract comprises
treating the bacterial infection with composition comprising an
effective amount of at least one lytic enzyme produced by a
bacteria infected with a bacteriophage specific for the bacteria,
and a carrier for delivering said lytic enzyme to the digestive
tract. In a preferred embodiment of the invention, the bacterial
infections being treated are being caused by gram negative bacteria
selected from the group consisting of Listeria, Salmonella, E.
coli, and Campylobacter. However, this method and composition will
effectively treat other bacteria, when the appropriate lytic enzyme
is used. The lytic enzymes used in the digestive tract may be
either supplemented by chimeric and/or shuffled lytic enzymes, or
may be itself a chimeric and/or shuffled lytic enzyme. Similarly, a
holin enzyme may be included, which may also be a chimeric and/or
shuffled lytic enzyme.
[0064] In a preferred embodiment of the invention, the carrier is
selected from the group consisting of suppository enemas, syrups,
or enteric coated pills. These proposed carriers can be made by
conventional methods. However, the only difference in their
manufacture is that the enzyme being placed in the carrier must not
be allowed to denature. To that end, the enzyme should be
incorporated into a carrier which does not contain alcohol, and
which has been cooled to a temperature that will not cause the
denaturing of the enzyme. The enzyme may be incorporated in a
lyophilized state, or may be incorporated in a liposome before
being placed in the suppository, syrup or enteric coated pill.
[0065] The enzyme placed in the composition or carrier should be in
an environment having a pH which allows for activity of the lytic
enzyme. To this end, the pH of the composition is preferably kept
in a range of between about 2 and about 11, more preferably in a
range of between about between about 4.0 and about 9.0, and even
more preferably at a pH range of between about 5.5 and about 7.5.
As described above with the other lytic enzyme, the pH can be
moderated by the use of a buffer. The buffer may contain a reducing
agent, and more specifically dithiothreitol. The buffer may also be
a a metal chelating reagent, such as ethylenediaminetetracetic
disodium salt or the buffer may contain a citrate-phosphate buffer.
As with all compositions described in this patent, the composition
may, further include a bactericidal or bacteriostatic agent as a
preservative.
[0066] The lytic enzyme is preferably present in a concentration of
about 100 to about 500,000 active enzyme units per milliliter of
fluid in the wet environment of a mucous membrane surface or the
gastrointestinal, preferably about 100 to about 100,000 active
enzyme units per milliliter of fluid, and preferably present in a
concentration of about 100 to about 10,000 active enzyme units per
milliliter of fluid in the wet environment of the a mucous membrane
surface and/orgastrointestinal tract.
[0067] The suppository is known in the art, and is made of
glycerin, fatty acids, and similar type substances that dissolve at
body temperature. As the suppository dissolves, the phage
associated lytic enzyme will be released.
[0068] Another composition and use of the lytic enzyme is for the
therapeutic or prophylactic treatment of bacterial infections of
burns and wounds of the skin. The composition comprises an
effective amount of at least one lytic enzyme produced by a
bacteria infected with a bacteriophage specific for the bacteria
and a carrier for delivering at least one lytic enzyme to the
wounded skin. The lytic enzyme(s) used for the topical treatment of
burns may be either supplemented by chimeric and/or shuffled lytic
enzymes, or may be itself a chimeric and/or shuffled lytic enzyme.
Similarly, a holin enzyme may be included, which may also be a
chimeric and/or shuffled lytic enzyme. The mode of application for
the lytic enzyme includes a number of different types and
combinations of carriers which include, but are not limited to an
aqueous liquid, an alcohol base liquid, a water soluble gel, a
lotion, an ointment, a nonaqueous liquid base, a mineral oil base,
a blend of mineral oil and petrolatum, lanolin, liposomes, protein
carriers such as serum albumin or gelatin, powdered cellulose
carmel, and combinations thereof. A mode of delivery of the carrier
containing the therapeutic agent includes but is not limited to a
smear, spray, a time-release patch, a liquid absorbed wipe, and
combinations thereof. The lytic enzyme may be applied to a bandage
either directly or in one of the other carriers. The bandages may
be sold damp or dry, wherein the enzyme is in a lyophilized form on
the bandage. This method of application is most effective for the
treatment of burns.
[0069] The carriers of the compositions of the present invention
may comprise semi-solid and gel-like vehicles that include a
polymer thickener, water, preservatives, active surfactants or
emulsifiers, antioxidants, sun screens, and a solvent or mixed
solvent system. U.S. Pat. No. 5,863,560 (Osborne) discusses a
number of different carrier combinations which can aid in the
exposure of the skin to a medicament.
[0070] Polymer thickeners that may be used include those known to
one skilled in the art, such as hydrophilic and hydroalcoholic
gelling agents frequently used in the cosmetic and pharmaceutical
industries. Preferably, the hydrophilic or hydroalcoholic gelling
agent comprises "CARBOPOL.RTM." (B.F. Goodrich, Cleveland, Ohio),
"HYPAN.RTM." (Kingston Technologies, Dayton, N.J.), "NATROSOL.RTM."
(Aqualon, Wilmington, Del.), "KLUCEL.RTM." (Aqualon, Wilmington,
Del.), or "STABILEZE.RTM." (ISP Technologies, Wayne, N.J.).
Preferably, the gelling agent comprises between about 0.2% to about
4% by weight of the composition. More particularly, the preferred
compositional weight percent range for "CARBOPOL.RTM." is between
about 0.5% to about 2%, while the preferred weight percent range
for "NATROSOL.RTM." and "KLUCEL.RTM." is between about 0.5% to
about 4%. The preferred compositional weight percent range for both
"HYPAN.RTM." and "STABILEZE.RTM." is between about 0.5% to about
4%.
[0071] "CARBOPOL.RTM." is one of numerous cross-linked acrylic acid
polymers that are given the general adopted name carbomer. These
polymers dissolve in water and form a clear or slightly hazy gel
upon neutralization with a caustic material such as sodium
hydroxide, potassium hydroxide, triethanolamine, or other amine
bases. "KLUCEL.RTM." is a cellulose polymer that is dispersed in
water and forms a uniform gel upon complete hydration. Other
preferred gelling polymers include hydroxyethylcellulose, cellulose
gum, MVE/MA decadiene crosspolymer, PVM/MA copolymer, or a
combination thereof.
[0072] Preservatives may also be used in this invention and
preferably comprise about 0.05% to 0.5% by weight of the total
composition. The use of preservatives assures that if the product
is microbially contaminated, the formulation will prevent or
diminish microorganism growth. Some preservatives useful in this
invention include methylparaben, propylparaben, butylparaben,
chloroxylenol, sodium benzoate, DMDM Hydantoin,
3-Iodo-2-Propylbutyl carbamate, potassium sorbate, chlorhexidine
digluconate, or a combination thereof.
[0073] Titanium dioxide may be used as a sunscreen to serve as
prophylaxis against photosensitization. Alternative sun screens
include methyl cinnamate. Moreover, BHA may be used as an
antioxidant, as well as to protect ethoxydiglycol and/or dapsone
from discoloration due to oxidation. An alternate antioxidant is
BHT.
[0074] Pharmaceuticals for use in all embodiments of the invention
include antimicrobial agents, anti-inflammatory agents, antiviral
agents, local anesthetic agents, corticosteroids, destructive
therapy agents, antifungals, and antiandrogens. In the treatment of
acne, active pharmaceuticals that may be used include antimicrobial
agents, especially those having anti-inflammatory properties such
as dapsone, erythromycin, minocycline, tetracycline, clindamycin,
and other antimicrobials. The preferred weight percentages for the
antimicrobials are 0.5% to 10%.
[0075] Local anesthetics include tetracaine, tetracaine
hydrochloride, lidocaine, lidocaine hydrochloride, dyclonine,
dyclonine hydrochloride, dimethisoquin hydrochloride, dibucaine,
dibucaine hydrochloride, butambenpicrate, and pramoxine
hydrochloride. A preferred concentration for local anesthetics is
about 0.025% to 5% by weight of the total composition. Anesthetics
such as benzocaine may also be used at a preferred concentration of
about 2% to 25% by weight.
[0076] Corticosteroids that may be used include betqamethasone
dipropionate, fiuocinolone actinide, betamethasone valerate,
triamcinolone actinide, clobetasol propionate, desoximetasone,
diflorasone diacetate, amcinonide, flurandrenolide, hydrocortisone
valerate, hydrocortisone butyrate, and desonide are recommended at
concentrations of about 0.01% to 1.0% by weight. Preferred
concentrations for corticosteroids such as hydrocortisone or
methylprednisolone acetate are from about 0.2% to about 5.0% by
weight.
[0077] Destructive therapy agents such as salicylic acid or lactic
acid may also be used. A concentration of about 2% to about 40% by
weight is preferred. Cantharidin is preferably utilized in a
concentration of about 5% to about 30% by weight. Typical
antifungals that may be used in this invention and their preferred
weight concentrations include: oxiconazole nitrate (0.1% to 5.0%),
ciclopirox olamine (0.1% to 5.0%), ketoconazole (0.1% to 5.0%),
miconazole nitrate (0.1% to 5.0%), and butoconazole nitrate (0.1%
to 5.0%). For the topical treatment of seborrheic dermatitis,
hirsutism, acne, and alopecia, the active pharmaceutical may
include an antiandrogen such as flutamide or finasteride in
preferred weight percentages of about 0.5% to 10%.
[0078] Typically, treatments using a combination of drugs include
antibiotics in combination with local anesthetics such as polymycin
B sulfate and neomycin sulfate in combination with tetracaine for
topical antibiotic gels to provide prophylaxis against infection
and relief of pain. Another example is the use of minoxidil in
combination with a corticosteroid such as betamethasone
diproprionate for the treatment of alopecia ereata. The combination
of an anti-inflammatory such as cortisone with an antifungal such
as ketoconazole for the treatment of tinea infections is also an
example.
[0079] In one embodiment, the invention comprises a dermatological
composition having about 0.5% to 10% carbomer and about 0.5% to 10%
of a pharmaceutical that exists in both a dissolved state and a
micro particulate state. The dissolved pharmaceutical has tne
capacity to cross the stratum corneum, whereas the micro
particulate pharmaceutical does not. Addition of an amine base,
potassium, hydroxide solution, or sodium hydroxide solution
completes the formation of the gel. More particularly, the
pharmaceutical may include dapsone, an antimicrobial agent having
anti-inflammatory properties. A preferred ratio of micro
particulate to dissolved dapsone is five or less.
[0080] In another embodiment, the invention comprises about 1%
carbomer, about 80-90% water, about 10% ethoxydiglycol, about 0.2%
methylparaben, about 0.3% to 3.0% dapsone including both micro
particulate dapsone and dissolved dapsone, and about 2% caustic
material. More particularly, the carbomer may include
"CARBOPOL.RTM. 980" and the caustic material may include sodium
hydroxide solution.
[0081] In a preferred embodiment, the composition comprises dapsone
and ethoxydiglycol, which allows for an optimized ratio of micro
particulate drug to dissolved drug. This ratio determines the
amount of drug delivered, compared to the amount of drug retained
in or above the stratum corneum to function in the supracorneum
domain. The system of dapsone and ethoxydiglycol may include
purified water combined with "CARBOPOL.RTM." gelling polymer,
methylparaben, propylparaben, titanium dioxide, BHA, and a caustic
material to neutralize the "CARBOPOL.RTM."
[0082] Any of the carriers for the lytic enzyme may be manufactured
by conventional means. However, if alcohol is used in the carrier,
the enzyme should be in a micelle, liposome, or a "reverse"
liposome, to prevent denaturing of the enzyme. Similarly, when the
lytic enzyme is being placed in the carrier, and the carrier is, or
has been heated, such placement should be made after the carrier
has cooled somewhat, to avoid heat denaturation of the enzyme. In a
preferred embodiment of the invention, the carrier is sterile.
[0083] The enzyme may be added to these substances in a liquid form
or in a lyophilized state, whereupon it will be solubilized when it
meets a liquid body.
[0084] The effective dosage rates or amounts of the lytic enzyme to
treat the infection, and the duration of treatment will depend in
part on the seriousness of the infection, the duration of exposure
of the recipient to the infectious bacteria, the number of square
centimeters of skin or tissue which are infected, the depth of the
infection, the seriousness of the infection, and a variety of a
number of other variables. The composition may be applied anywhere
from once to several times a day, and may be applied for a short or
long term period. The usage may last for days or weeks. Any dosage
form employed should provide for a minimum number of units for a
minimum amount of time. The concentration of the active units of
enzyme believed to provide for an effective dosage may be in the
range of about 100 units/ml to about 500,000 units/ml of enzyme,
preferably in the range of about 1000 units/ml to about 100,000
units/ml, and most preferably from about 10,000 to 100,000
units/ml. The amount of active units per ml and the duration of
time of exposure depends on the nature of infection, and the amount
of contact the carrier allows the lytic enzyme to have. It is to be
remembered that the enzyme works best when in a fluid environment.
Hence, effectiveness of the enzyme is in part related to the amount
of moisture trapped by the carrier. In another preferred
embodiment, a mild surfactant in an amount effective to potentiate
the therapeutic effect of the lytic enzyme. Suitable mild
surfactants include, inter alia, esters of polyoxyethylene sorbitan
and fatty acids (Tween series), octylphenoxy polyethoxy ethanol
(Triton-X series), n-Octyl-.beta.-D-glucopyranoside,
n-Octyl-.beta.-D-thioglucopyranoside,
n-Decyl-.beta.-D-glucopyranoside,
n-Dodecyl-.beta.-D-glucopyranoside, and biologically occurring
surfactants, e.g., fatty acids, glycerides, monoglycerides,
deoxycholate and esters of deoxycholate.
[0085] In order to accelerate treatment of the infection, the
therapeutic agent may further include at least one complementary
agent which can also potentiate the bactericidal activity of the
lytic enzyme. The complementary agent can be penicillin, synthetic
penicillins bacitracin, methicillin, cephalosporin, polymyxin,
cefaclor. Cefadroxil, cefamandole nafate, cefazolin, cefixime,
cefmetazole, cefonioid, cefoperazone, ceforanide, cefotanme,
cefotaxime, cefotetan, cefoxitin, cefpodoxime proxetil,
ceftazidime, ceftizoxime, ceftriaxone, cefriaxone moxalactam,
cefuroxime, cephalexin, cephalosporin C, cephalosporin C sodium
salt, cephalothin, cephalothin sodium salt, cephapirin, cephradine,
cefuroximeaxetil, dihydratecephalothin, moxalactam, loracarbef.
mafate, chelating agents and any combinations thereof in amounts
which are effective to synergistically enhance the therapeutic
effect of the lytic enzyme.
[0086] Additionally, the therapeutic agent may further comprise the
enzyme lysostaphin for the treatment of any Staphylococcus aureus
bacteria. Mucolytic peptides, such as lysostaphin, have been
suggested to be efficacious in the treatment of S. aureus
infections of humans (Schaffner et al., Yale J. Biol. & Med.,
39:230 (1967) and bovine mastitis caused by S. aureus (Sears et
al., J. Dairy Science, 71 (Suppl. 1): 244(1988)). Lysostaphin, a
gene product of Staphylococcus simulans, exerts a bacteriostatic
and bactericidal effect upon S. aureus by enzymatically degrading
the polyglycine crosslinks of the cell wall (Browder et al., Res.
Comm., 19: 393-400 (1965)). U.S. Pat. No. 3,278,378 describes
fermentation methods for producing lysostaphin from culture media
of S. staphylolyticus, later renamed S. simulans. Other methods for
producing lysostaphin are further described in U.S. Pat. Nos.
3,398,056 and 3,594,284. The gene for lysostaphin has subsequently
been cloned and sequenced (Recsei et al., Proc. Natl. Acad. Sci.
USA, 84: 1127-1131 (1987)). The recombinant mucolytic bactericidal
protein, such as r-lysostaphin, can potentially circumvent problems
associated with current antibiotic therapy because of its targeted
specificity, low toxicity and possible reduction of biologically
active residues. Furthermore, lysostaphin is also active against
non-dividing cells, while most antibiotics require actively
dividing cells to mediate their effects (Dixon et al., Yale J.
Biology and Medicine, 41: 62-68 (1968)). Lysostaphin, in
combination with the lysin enzyme, can be used in the presence or
absence of the listed antibiotics. There is a degree of added
importance in using both lysostaphin and the lysin enzyme in the
same therapeutic agent. Frequently, when a body has a bacterial
infection, the infection by one genus of bacteria weakens the body
or changes the bacterial flora of the body, allowing other
potentially pathogenic bacteria to infect the body. One of the
bacteria that sometimes co-infects a body is Staphylococcus aureus.
Many strains of Staphylococcus aureus produce penicillinase, such
that Staphylococcus, Streptococcus, and other gram positive
bacterial strains will not be killed by standard antibiotics.
Consequently, the use of the lysin and lysostaphin, possibly in
combination with antibiotics, can serve as the most rapid and
effective treatment of bacterial infections. In yet another
preferred embodiment, the invention may include mutanolysin, and
lysozyme
[0087] In preferred embodiments of the invention, the lytic enzymes
for Pseudomonas, Staphylococcus, and Streptococcus, jointly or
individually, may be incorporated into the carrier, or into a
bandage to be used on burn patients, or in a solution or cream
carrier.
[0088] Yet another use of lytic enzymes is for the prophylactic or
therapeutic treatment of vaginal infections. This treatment
comprises treating the vaginal infection with an effective amount
of at least one lytic enzyme produced by a bacteria being infected
with a bacteriophage specific for that bacteria, wherein that lytic
enzyme is incorporated in a carrier to be placed in a vagina. The
lytic enzyme(s) used to treat bacterial infections of the vagina
may be either supplemented by chimeric and/or shuffled lytic
enzymes, or may be itself a chimeric and/or shuffled lytic enzyme.
Similarly, a holin enzyme may be included, which may also be a
chimeric and/or shuffled lytic enzyme. The preferred carrier is a
tampon, or vaginal douche. A pad may also be used as a carrier,
although it is not as effective. While any number of bacteria could
be treated using this composition and method, it is believed that
the most optimum use of this treatment composition and method would
be for the treatment of an E. coli and Streptococcus B infection.
Vaginal infections caused by Group B Streptococcus can cause
neonatal meningitis resulting in brain damage and premature death.
Lytic enzyme incorporated into tampon specific for group B Strep
would eliminate the group B organisms without disturbing normal
flora so that woman would not be overcome by yeast infection post
antibiotic therapy. The use of the lytic enzyme in the vagina would
best provide a prophylactic effect, although therapeutic use would
also be advisable.
[0089] To produce a pad or tampon containing the enzyme, the lytic
enzyme can be applied in a solution to the tampon, and allowed to
dry. The lytic enzyme may be incorporated into the pad or tampon by
any other means known in the art, including lyophilization,
spraying, etc. The tampons and pads may also be kept slightly
moist, and in a sealed wrapper until ready for use. In that case,
bactericide and bacteriostatic compounds and inhibitors should be
present in the tampons and pads. The method to be used for
incorporating the lytic enzyme into the tampon or pad can be one of
the methods known in the art for incorporating a pharmaceutical
product. In another embodiment of the invention, the lytic enzyme
is incorporated into a vaginal suppository. The vaginal suppository
into which the lytic enzyme is being incorporated may be a standard
vaginal suppository, comprised of glyceride, alginate, starch,
other standard binders and any combinations thereof.
[0090] When using a tampon as the carrier, it is best to insert the
tampon in the vagina and leave it in for up to 12 hours to
distribute the enzyme vaginally.
[0091] As with other lytic enzymes, it is preferable that the pH be
kept in a range of about 4.0 and about 9.0 even more preferably at
a pH range of between about 5.5 and about 7.5. As described above
with the other lytic enzyme, the pH can be moderated by the use of
a buffer. The buffer may contain a reducing agent, and more
specifically dithiothreitol. The buffer may also contain a metal
chelating reagent, such as ethylenediaminetetracetic disodium salt
or the buffer may be a citrate-phosphate buffer. As with all
compositions described in this patent, the composition may, further
include a bactericidal or bacteriostatic agent as a
preservative.
[0092] The lytic enzyme is preferably present in a concentration of
about 100 to about 500,000 active enzyme units per milliliter of
fluid in the wet environment of the vaginal tract, preferably about
100 to about 100,000 active enzyme units per milliliter of fluid,
and preferably present in a concentration of about 100 to about
10,000 active enzyme units per milliliter of fluid in the wet
environment of the vaginal tract.
[0093] Another use of the invention is for the prophylactic and
therapeutic treatment of eye infections. The method of treatment
comprises administering eye drops which comprise an effective
amount of at least one lytic enzyme produced by the bacteria being
infected with a bacteriophage specific for the bacteria and a
carrier capable of being safely applied to an eye, with the carrier
containing the lytic enzyme. In a preferred embodiment of the
invention, the bacteria being treated is Hemophilus or
Staphylococcus. The eye drops are in the form of an isotonic
solution. The pH of the solution should be adjusted so that there
is no irritation of the eye, which in turn would lead to possibly
infection by other organisms, and possibly to damage to the eye.
While the pH range should be in the same range as for other lytic
enzymes, the most optimal pH will be in the range of from 6.0 to
7.5. Similarly, buffers of the sort described above for the other
lytic enzymes should also be used. Other antibiotics which are
suitable for use in eye drops may be added to the composition
containing the lytic enzymes. Bactericides and bacteriostatic
compounds may also be added. As stated above, this lytic enzyme may
be either supplemented by chimeric and/or shuffled lytic enzymes,
or may be itself a chimeric and/or shuffled lytic enzyme.
Similarly, a holin enzyme may be included, which may also be a
chimeric and/or shuffled lytic enzyme.
[0094] It is to be remembered that all of the enzymes can be used
for prophylactic and therapeutic treatments of the bacteria for
which the enzymes are specific.
[0095] It is also to be remembered that a carrier may have more
than one lytic enzyme. For instance, A throat lozenge may comprise
just a lysin enzyme (which lyses the Streptococcus A strain causing
"strep" throat, or it may also include the lytic enzymes for
Hemophilus. Similarly, the carrier for treating burns and wounds,
or infections of the skin, may contain just one lytic enzyme, or a
combination of lytic enzymes, for the treatment of Pseudomonas,
Streptococcus, Staphylococcus, or any other of a number of
bacteria.
[0096] Lytic enzymes can also be used to fight dental caries.
Specifically, a lytic enzyme specific for Streptococcus mutans may
be incorporated in a toothpaste or oral wash. Similarly, this lytic
enzyme may also be incorporated into a chewing gum or lozenge. Any
other carrier can be used that allows for the exposure of the
mouth, gums, and teeth to the lytic enzyme.
[0097] The lytic enzyme may also be incorporated in a lyophilized
or dried form in tooth powder. If the lytic enzyme is to be used in
an oral wash, it is preferred that the oral wash not contain any
alcohol, so as to not denature the enzyme. The enzyme can also be
in a liposome when mixed in with the toothpaste or oral wash. The
concentrations of the enzyme units per ml of toothpaste or mouth
wash can be in the range of from about 100 units/ml to about
500,000 units/ml of composition, preferably in the range of about
1000 units/ml to about 100,000 units/ml, and most preferably from
about 10,000 to 100,000 units/ml. The pH of the toothpaste or oral
wash should be in a range that allows for the optimum performance
of the enzyme, while not causing any discomfort to the user of the
toothpaste or oral wash. Again, as with the other uses of lytic
enzymes, the lytic enzyme use to treat dental caries may be either
supplemented by chimeric and/or shuffled lytic enzymes, or may be
itself a chimeric and/or shuffled lytic enzyme. Similarly, a holin
enzyme may be included, which may also be a chimeric and/or
shuffled lytic enzyme.
[0098] Many modifications and variations of the present invention
are possible in light of the above teachings. It is, therefore, to
be understood within the scope of the appended claims the invention
may be protected otherwise than as specifically described.
* * * * *