U.S. patent application number 17/361123 was filed with the patent office on 2021-12-23 for implantable medical devices with increased immune tolerance, and methods for making and implanting.
The applicant listed for this patent is AMPIO PHARMACEUTICALS, INC.. Invention is credited to David Bar-Or.
Application Number | 20210393851 17/361123 |
Document ID | / |
Family ID | 1000005826245 |
Filed Date | 2021-12-23 |
United States Patent
Application |
20210393851 |
Kind Code |
A1 |
Bar-Or; David |
December 23, 2021 |
IMPLANTABLE MEDICAL DEVICES WITH INCREASED IMMUNE TOLERANCE, AND
METHODS FOR MAKING AND IMPLANTING
Abstract
The present invention relates to the contacting of one or more
surfaces of an implantable medical device with one or more
diketopiperazines (DKPs).
Inventors: |
Bar-Or; David; (Englewood,
CO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AMPIO PHARMACEUTICALS, INC. |
Englewood |
CO |
US |
|
|
Family ID: |
1000005826245 |
Appl. No.: |
17/361123 |
Filed: |
June 28, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16575653 |
Sep 19, 2019 |
11058798 |
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17361123 |
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15896964 |
Feb 14, 2018 |
10471178 |
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16575653 |
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14350617 |
Apr 9, 2014 |
9925300 |
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PCT/US12/59483 |
Oct 10, 2012 |
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15896964 |
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61545465 |
Oct 10, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61L 27/28 20130101;
A61L 29/16 20130101; A61L 2300/606 20130101; A61K 31/495 20130101;
A61L 31/16 20130101; A61L 2300/204 20130101; F04C 2270/0421
20130101; A61L 27/54 20130101; A61K 31/496 20130101; A61L 2300/41
20130101 |
International
Class: |
A61L 27/28 20060101
A61L027/28; A61L 29/16 20060101 A61L029/16; A61L 31/16 20060101
A61L031/16; A61K 31/495 20060101 A61K031/495; A61K 31/496 20060101
A61K031/496; A61L 27/54 20060101 A61L027/54 |
Claims
1-87. (canceled)
88. A medical device coated with DA-DKP and a component selected
from the group consisting of N-acetyl tryptophan, caprylate,
caprylic acid, and combinations thereof.
89. The medical device of claim 88, wherein the component is
N-acetyl tryptophan.
90. The medical device of claim 88, wherein the component is
caprylate, caprylic acid, or both caprylate and caprylic acid.
91. The medical device of claim 88, wherein the component is
N-acetyl tryptophan and caprylate.
92. The medical device of claim 88, wherein the component is
N-acetyl tryptophan and caprylic acid.
93. The medical device of claim 88, wherein the component is
N-acetyl tryptophan, caprylate and caprylic acid.
94. The medical device of claim 88, which is implantable.
95. The medical device of claim 88, which is selected from the
group consisting of a graft, catheter, stent, prosthetic, breast
implant, pump, tube, pin, rod, screw, brace, plate and
pacemaker.
96. The medical device of claim 88, which is a tracheal tube.
97. The medical device of claim 96, which is an endotracheal
tube.
98. A method of preparing a medical device, comprising contacting
the medical device with DA-DKP and a component selected from the
group consisting of N-acetyl tryptophan, caprylate, caprylic acid,
and combinations thereof.
99. The method of claim 98, wherein the component is N-acetyl
tryptophan.
100. The method of claim 98, wherein the component is caprylate,
caprylic acid, or both caprylate and caprylic acid.
101. The method of claim 98, wherein the component is N-acetyl
tryptophan and caprylate or N-acetyl tryptophan and caprylic
acid.
102. The method of claim 98, wherein the device is implantable.
103. The method of claim 98, wherein the device is selected from
the group consisting of a graft, catheter, stent, prosthetic,
breast implant, pump, tube, pin, rod, screw, brace, plate and
pacemaker.
104. The method of claim 98, wherein the device is a tracheal
tube.
105. The method of claim 104, wherein the device is an endotracheal
tube.
106. The method of claim 98, wherein the DA-DKP is coated onto the
surface of the device.
107. A method for implanting a medical device into a subject,
wherein a surface of the device is coated with DA-DKP, N-acetyl
tryptophan, caprylate, caprylic acid, or combinations thereof.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 16/575,653, filed Sep. 19, 2019; which is a
continuation of U.S. patent application Ser. No. 15/896,964, filed
Feb. 14, 2018, now U.S. Pat. No. 10,471,178; which is a
continuation of U.S. patent application Ser. No. 14/350,617, filed
Apr. 9, 2014, now U.S. Pat. No. 9,925,300; which is a national
stage application under 35 U.S.C. 371 of PCT Application No.
PCT/US2012/059483, having an international filing date of Oct. 10,
2012, which designated the United States; which claims the benefit
of priority under 35 U.S.C. .sctn. 119(e) to U.S. Provisional
Patent Application No. 61/545,465, filed Oct. 10, 2011. The entire
disclosures of U.S. Provisional Patent Application No. 61/545,465,
U.S. patent application Ser. No. 14/350,617, U.S. patent
application Ser. No. 15/896,964, U.S. patent application Ser. No.
16/575,653, and PCT Application No. PCT/US2012/059483 are
incorporated herein by reference in their entirety.
FIELD OF INVENTION
[0002] The present invention relates to the contacting one or more
surfaces of an implantable medical device with one or more
diketopiperazines (DKPs).
BACKGROUND OF THE INVENTION
[0003] Implantation of a medical device into a patient's body can
trigger an abnormal immune response by the patient's body which is
a threat to the acceptance of the implant and can result in device
failure. For the patient, this can mean extended inflammation,
higher risk of infection, and tissue build-up that may cause
complications as well as discomfort. These effects slow patient
recovery and often mean further medical intervention.
SUMMARY OF THE INVENTION
[0004] The present invention provides for a method to prepare an
implantable medical device for implantation in a subject comprising
contacting the device with a DKP.
[0005] The invention further provides for a method for implanting a
medical device comprising implanting a medical device into a
subject, wherein a surface of the device comprises a DKP.
[0006] In one aspect, the methods comprise a DKP comprising the
following formula
##STR00001##
wherein:
[0007] R.sup.1 and R.sup.2, which may be the same or different,
each is: [0008] (a) a side chain of an amino acid, wherein the
amino acid is glycine, alanine, valine, norvaline,
.alpha.-aminoisobutyric acid, 2,4-diaminobutyric acid,
2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine,
homoserine, threonine, aspartic acid, asparagine, glutamic acid,
glutamine, lysine, hydroxylysine, histidine, arginine,
homoarginine, citrulline, phenylalanine, p-aminophenylalanine,
tyrosine, tryptophan, thyroxine, cysteine, homocysteine,
methionine, penicillamine or ornithine; provided, however, that
when R.sup.1 is the side chain of asparagine or glutamine, then
R.sup.2 cannot be the side chain of lysine or ornithine, and when
R.sup.1 is the side chain of lysine or ornithine, then R.sup.2
cannot be the side chain of asparagine or glutamine; [0009] (b)
R.sup.1 is --CH.sub.2--CH.sub.2--CH.sub.2-- or
--CH.sub.2--CH(OH)--CH.sub.2-- and together with the adjacent ring
nitrogen forms proline or hydroxyproline, R.sup.2 is
--CH.sub.2--CH.sub.2--CH.sub.2-- or --CH.sub.2--CH(OH)--CH.sub.2--
and together with the adjacent ring nitrogen forms proline or
hydroxyproline, or both R.sup.1 and R.sup.2 are each independently
--CH.sub.2--CH.sub.2--CH.sub.2-- or --CH.sub.2--CH(OH)--CH.sub.2--
and together with the adjacent ring nitrogens form proline or
hydroxyproline; or [0010] (c) a derivative of a side chain of an
amino acid, wherein the amino acid is one of those recited in (a),
and the derivatized side chain has: [0011] (i) an --NH.sub.2 group
replaced by an --NHR.sup.3 or --N(R.sup.3).sub.2 group, wherein
each R.sup.3 may independently be a substituted or unsubstituted
alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0012] (ii) and --OH group replaced by an
--O--PO.sub.3H.sub.2 or --OR.sup.3 group, wherein each R.sup.3 may
independently be a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0013]
(iii) a --COOH group replaced by a --COOR.sup.3 group, wherein each
R.sup.3 may independently be a substituted or unsubstituted alkyl,
cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0014] (iv) a --COOH group replaced by a
--CON(R.sup.4).sub.2 group, wherein each R.sup.4 may independently
be H or a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0015]
(v) an --SH group replaced by --S--S--CH.sub.2--CH(NH.sub.2)--COOH
or --S--S--CH.sub.2--CH.sub.2--CH(NH.sub.2)--COOH; [0016] (vi) a
--CH.sub.2-- group replaced by a --CH(NH.sub.2)-- or a --CH(OH)--
group; [0017] (vii) a --CH.sub.3 group replaced by a
--CH.sub.2--NH.sub.2 or a --CH.sub.2--OH group; and/or [0018]
(viii) an H which is attached to a carbon atom replaced by a
halogen; or a physiologically-acceptable salt thereof.
[0019] The implantable medical device can be selected from a graft,
catheter, stent, prosthetic, breast implant, pump, tube, pin, rod,
screw, brace, plate and pacemaker. In one aspect, the stent can be
selected from a cardiac stent, an artery stent and a birth control
stent. In another aspect, the prosthetic can be selected from an
artificial hip, an artificial knee and an artificial ankle. In
still another aspect, the pump can be an insulin pump. In yet
another aspect, the implantable medical device is composed of
material selected from metal, steal, titanium, glass, polymers,
plastics and ceramics.
[0020] In one aspect, the DKP is adhered to the surface of the
implantable medical device. In another aspect, the DKP is
impregnated into the surface of the implantable medical device. In
still another aspect, the DKP is coated onto the surface of the
implantable medical device. In yet another aspect, the implantable
medical device is contacted with a solution comprising a DKP,
wherein the solution adheres to the device.
[0021] In some embodiments, the DKP that adheres to the surface of
the implantable medical device, or is impregnated into the surface
of the implantable medical device or coated onto the surface of the
implantable medical device, can be in an amount of about 1 .mu.M to
about 500 .mu.M. In still another aspect, the amount can be in an
amount of about 50 .mu.M to about 100 .mu.M.
[0022] In some embodiments, the concentration of the DKP on a
surface of the implantable medical device exposed to the body is
about 1 ng/cm.sup.2 to about 200 ng/cm.sup.2. In another aspect,
the concentration of the DKP on a surface of the implantable
medical device exposed to the body is about 50 ng/cm.sup.2.
[0023] The subject of the methods can be a mammal, including a
human.
[0024] In some embodiments, the subject's immune tolerance to the
device is increased by the presence of the DKP.
[0025] Another embodiment of the invention relates to an
implantable medical device, wherein a surface of the device
comprises a DKP. In one aspect, the device comprises a DKP
comprising the following formula
##STR00002##
wherein:
[0026] R.sup.1 and R.sup.2, which may be the same or different,
each is: [0027] (a) a side chain of an amino acid, wherein the
amino acid is glycine, alanine, valine, norvaline,
.alpha.-aminoisobutyric acid, 2,4-diaminobutyric acid,
2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine,
homoserine, threonine, aspartic acid, asparagine, glutamic acid,
glutamine, lysine, hydroxylysine, histidine, arginine,
homoarginine, citrulline, phenylalanine, p-aminophenylalanine,
tyrosine, tryptophan, thyroxine, cysteine, homocysteine,
methionine, penicillamine or ornithine; provided, however, that
when R.sup.1 is the side chain of asparagine or glutamine, then
R.sup.2 cannot be the side chain of lysine or ornithine, and when
R.sup.1 is the side chain of lysine or ornithine, then R.sup.2
cannot be the side chain of asparagine or glutamine; [0028] (b)
R.sup.1 is --CH.sub.2--CH.sub.2--CH.sub.2-- or
--CH.sub.2--CH(OH)--CH.sub.2-- and together with the adjacent ring
nitrogen forms proline or hydroxyproline, R.sup.2 is
--CH.sub.2--CH.sub.2--CH.sub.2-- or --CH.sub.2--CH(OH)--CH.sub.2--
and together with the adjacent ring nitrogen forms proline or
hydroxyproline, or both R.sup.1 and R.sup.2 are each independently
--CH.sub.2--CH.sub.2--CH.sub.2-- or --CH.sub.2--CH(OH)--CH.sub.2--
and together with the adjacent ring nitrogens form proline or
hydroxyproline; or [0029] (c) a derivative of a side chain of an
amino acid, wherein the amino acid is one of those recited in (a),
and the derivatized side chain has: [0030] (i) an --NH.sub.2 group
replaced by an --NHR.sup.3 or --N(R.sup.3).sub.2 group, wherein
each R.sup.3 may independently be a substituted or unsubstituted
alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0031] (ii) and --OH group replaced by an
--O--PO.sub.3H.sub.2 or --OR.sup.3 group, wherein each R.sup.3 may
independently be a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0032]
(iii) a --COOH group replaced by a --COOR.sup.3 group, wherein each
R.sup.3 may independently be a substituted or unsubstituted alkyl,
cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0033] (iv) a --COOH group replaced by a
--CON(R.sup.4).sub.2 group, wherein each R.sup.4 may independently
be H or a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0034]
(v) an --SH group replaced by --S--S--CH.sub.2--CH(NH.sub.2)--COOH
or --S--S--CH.sub.2--CH.sub.2--CH(NH.sub.2)--COOH; [0035] (vi) a
--CH.sub.2-- group replaced by a --CH(NH.sub.2)-- or a --CH(OH)--
group; [0036] (vii) a --CH.sub.3 group replaced by a
--CH.sub.2--NH.sub.2 or a --CH.sub.2--OH group; and/or [0037]
(viii) an H which is attached to a carbon atom replaced by a
halogen; or a physiologically-acceptable salt thereof.
[0038] The device can be selected from a graft, catheter, stent,
prosthetic, breast implant, pump, tube, pin, rod, screw, brace,
plate and pacemaker. In yet another aspect, the device is composed
of material selected from metal, steal, titanium, glass, polymers,
plastics and ceramics.
[0039] In one aspect, the DKP is adhered to the surface of the
device. In another aspect, the DKP is impregnated into the surface
of the device. In still another aspect, the DKP is coated onto the
surface of the device. In yet another aspect, the device is
contacted with a solution comprising a DKP, wherein the solution
adheres to the device.
[0040] In some embodiments, the DKP that adheres to the surface of
the device, or is impregnated into the surface of the device or
coated onto the surface of the device, can be in an amount of about
1 .mu.M to about 500 .mu.M. In still another aspect, the DKP amount
can be in an amount of about 50 .mu.M to about 100 .mu.M.
[0041] In some embodiments, the concentration of the DKP on a
surface of the device exposed to the body is about 1 ng/cm.sup.2 to
about 200 ng/cm.sup.2. In another aspect, the concentration of the
DKP on a surface of the device exposed to the body is about 50
ng/cm.sup.2.
DESCRIPTION OF THE INVENTION
[0042] The present invention relates to the contacting (such as by
coating, impregnating, etc.) of one or more surfaces of an
implantable medical device with one or more diketopiperazines
(DKPs). DKPs can suppress the immune response of a subject against
the implanted medical device and can be referred to as
immunomodulatory DKPs. By the introduction of DKPs to a surface of
an implanted device, the problems associated with immune response
against such devices (device failure and patient complications) can
be reduced. Without being bound by theory, by coating or contacting
an implantable medical device with a DKP, it is believed that the
presence of the DKP increases the subject's immune tolerance to the
device.
[0043] Various embodiments of the present invention include a
method to prepare an implantable medical device in a subject by
contacting the device with a DKP. Another embodiment is a method
for implanting a medical device by implanting a medical device into
a subject, wherein a surface of the device comprises a DKP. A
further embodiment of the present invention is an implantable
medical device, wherein a surface of the device comprises a
DKP.
[0044] To increase immune tolerance of an implantable medical
device and/or for implanting a medical device, the device and/or
the surface of the device can be contacted by the diketopiperazine
(DKP) of the present invention having the following formula:
##STR00003##
wherein:
[0045] R.sup.1 and R.sup.2, which may be the same or different,
each is: [0046] (a) a side chain of an amino acid, wherein the
amino acid is glycine, alanine, valine, norvaline,
.alpha.-aminoisobutyric acid, 2,4-diaminobutyric acid,
2,3-diaminobutyric acid, leucine, isoleucine, norleucine, serine,
homoserine, threonine, aspartic acid, asparagine, glutamic acid,
glutamine, lysine, hydroxylysine, histidine, arginine,
homoarginine, citrulline, phenylalanine, p-aminophenylalanine,
tyrosine, tryptophan, thyroxine, cysteine, homocysteine,
methionine, penicillamine or ornithine; provided, however, that
when R.sup.1 is the side chain of asparagine or glutamine, then
R.sup.2 cannot be the side chain of lysine or ornithine, and when
R.sup.1 is the side chain of lysine or ornithine, then R.sup.2
cannot be the side chain of asparagine or glutamine; [0047] (b)
R.sup.1 is --CH.sub.2--CH.sub.2--CH.sub.2-- or
--CH.sub.2--CH(OH)--CH.sub.2-- and together with the adjacent ring
nitrogen forms proline or hydroxyproline and/or R.sup.2 is
--CH.sub.2--CH.sub.2--CH.sub.2-- or --CH.sub.2--CH(OH)--CH.sub.2--
and together with the adjacent ring nitrogen forms proline or
hydroxyproline; or [0048] (c) a derivative of a side chain of an
amino acid, wherein the amino acid is one of those recited in (a),
and the derivatized side chain has: [0049] (i) an --NH.sub.2 group
replaced by an --NHR.sup.3 or --N(R.sup.3).sub.2 group, wherein
each R.sup.3 may independently be a substituted or unsubstituted
alkyl, cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0050] (ii) and --OH group replaced by an
--O--PO.sub.3H.sub.2 or --OR.sup.3 group, wherein each R.sup.3 may
independently be a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0051]
(iii) a --COOH group replaced by a --COOR.sup.3 group, wherein each
R.sup.3 may independently be a substituted or unsubstituted alkyl,
cycloalkyl, heterocycloalkyl, aryl, alkylaryl, arylalkyl or
heteroaryl; [0052] (iv) a --COOH group replaced by a
--CON(R.sup.4).sub.2 group, wherein each R.sup.4 may independently
be H or a substituted or unsubstituted alkyl, cycloalkyl,
heterocycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl; [0053]
(v) an --SH group replaced by --S--S--CH.sub.2--CH(NH.sub.2)--COOH
or --S--S--CH.sub.2--CH.sub.2--CH(NH.sub.2)--COOH; [0054] (vi) a
--CH.sub.2-- group replaced by a --CH(NH.sub.2)-- or a --CH(OH)--
group; [0055] (vii) a --CH.sub.3 group replaced by a
--CH.sub.2--NH.sub.2 or a --CH.sub.2--OH group; and/or [0056]
(viii) an H which is attached to a carbon atom replaced by a
halogen; or a physiologically-acceptable salt thereof.
[0057] By "replaced" is meant that, with reference to the formula
of an amino acid side chain, the specified group is replaced by the
other specified group. For instance, the formula of the isoleucine
side chain is --CH(CH.sub.3)--CH.sub.2--CH.sub.3. If the terminal
--CH.sub.3 group is replaced with a --CH.sub.2--OH group, then the
formula of the resulting derivatized isoleucine side chain would be
--CH(CH.sub.3)--CH.sub.2--CH.sub.2--OH. As another example, the
formula of the alanine side chain is --CH.sub.3. If one of the
hydrogen atoms is replaced by a chlorine atom, then the resulting
derivatized alanine side chain would be --CH.sub.2--Cl. Note that
the side chain of glycine is --H and, if this H is replaced by a
chlorine (or other halogen) atom, the resulting side chain will
--Cl, with the chlorine atom attached to the ring carbon (e.g.,
R.sup.1=--Cl)
[0058] Preferred are diketopiperazines wherein R.sup.1, R.sup.2 or
both is the side chain of aspartic acid or glutamic acid or a
derivative of such a side chain wherein the --COOH group is
replaced by a --COOR.sup.3 group or a --CON(R.sup.4).sub.2 group,
wherein R.sup.3 and R.sup.4 are defined above. Of this group of
compounds, most preferred are diketopiperazines comprising the side
chains of aspartic acid and alanine (Asp-Ala DKP or DA-DKP), the
side chains of glutamic acid and alanine (Glu-Ala DKP or EA-DKP),
the side chains of tyrosine and aspartic acid (Tyr-Asp DKP or
YD-DKP), the side chains of tyrosine and glutamic acid (Tyr-Glu DKP
or YE-DKP) and derivatives of the aspartic acid or glutamic acid
side chains of these four diketopiperazines wherein the --COOH
group is replaced by a --COOR.sup.3 group or a --CON(R.sup.4).sub.2
group, wherein R.sup.3 and R.sup.4 are defined above.
[0059] Also, preferred are diketopiperazines wherein R.sup.1 and
R.sup.2 are both hydrophobic side chains (e.g., the side chain of
phenylalanine) or hydrophobic side chain derivatives. By
"hydrophobic side chain derivative" is meant that the derivatized
side chain which is hydrophobic. In particular, preferred are
diketopiperzines wherein R.sup.1 and/or R.sup.2, which may be the
same or different, each is the side chain of glycine, alanine,
valine, norvaline, .alpha.-aminobutyric acid, leucine, isoleucine,
norleucine or phenylalanine, and/or R.sup.1 and/or R.sup.2 is
--CH.sub.2--CH.sub.2--CH.sub.2-- and together with the adjacent
nitrogen atom(s) form proline. Of this group of compounds, most
preferred are the diketopiperazines comprising the side chains of
glycine and leucine (Gly-Leu DKP or GL-DKP), proline and
phenylalanine (Pro-Phe DKP or PF-DKP), and alanine and proline
(Ala-Pro DKP or AP-DKP).
[0060] Additional preferred diketopiperazines are those wherein
R.sup.1, R.sup.2 or both is the side chain of methionine, the side
chain of arginine or a derivative of these side chains. Most
preferred of this group is a diketopiperazine wherein R.sup.1 is
the side chain of methionine and R.sup.2 is the side chain of
arginine (Met-Arg DKP or MR-DKP).
[0061] By "side chain" of an amino acid is meant that portion of
the amino acid attached to the common NH.sub.2--CH--COOH backbone
of all of the amino acids listed above. For instance, the side
chain of glycine is --H, the side chain of alanine is --CH.sub.3,
and the side chain of serine is --CH.sub.2OH.
[0062] By "hydrophobic" is meant a side chain or side chain
derivative that is uncharged at physiological pH and is repelled by
an aqueous solution.
[0063] By "alkyl" is meant a saturated straight-chain or branched
hydrocarbon containing 1-10 carbon atoms, preferably 1-6, carbon
atoms. "Lower alkyl" means a saturated straight-chain or branched
hydrocarbon containing 1-6 carbon atoms.
[0064] By "cycloalkyl" is meant a saturated cyclic hydrocarbon
containing at least one ring, each ring containing at least three
carbon atoms. Preferably, the cycloalkyl contains one ring of 4-8
carbon atoms.
[0065] By "heterocycloalkyl" is meant a cycloalkyl having one or
more of the ring carbon atoms of at least one of the rings replaced
by an O, S or N.
[0066] By "aryl" is meant an aromatic group having at least one
aromatic ring (e.g., phenyl).
[0067] By "alkylaryl" is meant a lower alkyl having an H replaced
by an aryl (e.g., --CH.sub.2--C.sub.6H.sub.5 or
--CH.sub.3CH(C.sub.6H.sub.5)CH.sub.3).
[0068] By "arylalkyl" is meant an aryl having an H replaced by a
lower alkyl (e.g., --C.sub.6H.sub.4--CH.sub.3).
[0069] By "heteroaryl" is meant an aryl having one or more of the
ring carbon atoms of at least one of the rings replaced by an O, S
or N.
[0070] By "substituted" is meant that the moiety is substituted
with one or more substituents selected from the following group:
--OH, NH.sub.2, --SH, --COOH and/or a halogen atom.
[0071] By "halogen" is meant chlorine, fluorine, bromine or iodine.
Preferred is chlorine or bromine.
[0072] Methods of making diketopiperazines are well known in the
art, and these methods may be employed to synthesize the
diketopiperazines of the invention. See, e.g., U.S. Pat. Nos.
4,694,081, 5,817,751, 5,990,112, 5,932,579 and 6,555,543, US Patent
Application Publication Number 2004/0024180, PCT applications WO
96/00391 and WO 97/48685, and Smith et al., Bioorg. Med. Chem.
Letters, 8, 2369-2374 (1998), the complete disclosures of which are
incorporated herein by reference.
[0073] For instance, diketopiperazines can be prepared by first
synthesizing dipeptides. The dipeptides can be synthesized by
methods well known in the art using L-amino acids, D-amino acids or
a combination of D- and L-amino acids. Preferred are solid-phase
peptide synthetic methods. Of course, dipeptides are also available
commercially from numerous sources, including DMI Synthesis Ltd.,
Cardiff, UK (custom synthesis), Sigma-Aldrich, St. Louis, Mo.
(primarily custom synthesis), Phoenix Pharmaceuticals, Inc.,
Belmont, Calif. (custom synthesis), Fisher Scientific (custom
synthesis) and Advanced ChemTech, Louisville, Ky.
[0074] Once the dipeptide is synthesized or purchased, it is
cyclized to form a diketopiperazine. This can be accomplished by a
variety of techniques. For example, U.S. Patent Application
Publication Number 2004/0024180 describes a method of cyclizing
dipeptides. Briefly, the dipeptide is heated in an organic solvent
while removing water by distillation. Preferably, the organic
solvent is a low-boiling azeotrope with water, such as
acetonitrile, allyl alcohol, benzene, benzyl alcohol, n-butanol,
2-butanol, t-butanol, acetic acid butylester, carbon tetrachloride,
chlorobenzene chloroform, cyclohexane, 1,2-dichlorethane,
diethylacetal, dimethylacetal, acetic acid ethylester, heptane,
methylisobutylketone, 3-pentanol, toluene and xylene. The
temperature depends on the reaction speed at which the cyclization
takes place and on the type of azeotroping agent used. The reaction
is preferably carried out at 50-200.degree. C., more preferably
80-150.degree. C. The pH range in which cyclization takes place can
be easily determined by the person skilled in the art. It will
advantageously be 2-9, preferably 3-7. When one or both of the
amino acids of the dipeptide has, or is derivatized to have, a
carboxyl group on its side chain (e.g., aspartic acid or glutamic
acid), the dipeptide is preferably cyclized as described in U.S.
Pat. No. 6,555,543. Briefly, the dipeptide, with the side-chain
carboxyl still protected, is heated under neutral conditions.
Typically, the dipeptide will be heated at from about 80.degree. C.
to about 180.degree. C., preferably at about 120.degree. C. The
solvent will be a neutral solvent. For instance, the solvent may
comprise an alcohol (such as butanol, methanol, ethanol, and higher
alcohols, but not phenol) and an azeotropic co-solvent (such as
toluene, benzene, or xylene). Preferably, the alcohol is
butan-2-ol, and the azeotropic co-solvent is toluene. The heating
is continued until the reaction is complete, and such times can be
determined empirically. Typically, the dipeptide will be cyclized
by refluxing it for about 8-24 hours, preferably about 18 hours.
Finally, the protecting group is removed from the diketopiperazine.
In doing so, the use of strong acids (mineral acids, such as
sulfuric or hydrochloric acids), strong bases (alkaline bases, such
as potassium hydroxide or sodium hydroxide), and strong reducing
agents (e.g., lithium aluminum hydride) should be avoided, in order
to maintain the chirality of the final compound.
[0075] Dipeptides made on solid phase resins can be cyclized and
released from the resin in one step. See, e.g., U.S. Pat. No.
5,817,751. For instance, the resin having an N-alkylated dipeptide
attached is suspended in toluene or toluene/ethanol in the presence
of acetic acid (e.g., 1%) or triethylamine (e.g., 4%). Typically,
basic cyclization conditions are preferred for their faster
cyclization times.
[0076] To prepare the diketopiperazine of formula I wherein the
amino acid side chains are derivatized, amino acid derivatives can
be used in the synthesis of the dipeptides, the dipeptides can be
derivatized and/or the diketopiperazines can be derivatized, as is
known in the art. See, e.g., those references cited above.
[0077] Other methods of cyclizing dipeptides and of making
diketopiperazines are known in the art and can be used in the
preparation of diketopiperazines useful in the practice of the
invention. See, e.g., those references listed above. In addition,
many diketopiperazines suitable for use in the present invention
can be made as described below from proteins and peptides. Further,
diketopiperazines for use in the practice of the invention can be
obtained commercially from, e.g., DMI Synthesis Ltd., Cardiff, UK
(custom synthesis).
[0078] The diketopiperazines of formula I include all possible
stereoisomers than can be obtained by varying the configuration of
the individual chiral centers, axes or surfaces. In other words,
the diketopierazines of formulas I and II include all possible
diastereomers, as well as all optical isomers (enantiomers).
[0079] The physiologically-acceptable salts of the
diketopiperazines of the invention may also be used in the practice
of the invention. Physiologically-acceptable salts include
conventional non-toxic salts, such as salts derived from inorganic
acids (such as hydrochloric, hydrobromic, sulfuric, phosphoric,
nitric, and the like), organic acids (such as acetic, propionic,
succinic, glycolic, stearic, lactic, malic, tartaric, citric,
glutamic, aspartic, benzoic, salicylic, oxalic, ascorbic acid, and
the like) or bases (such as the hydroxide, carbonate or bicarbonate
of a pharmaceutically-acceptable metal cation or organic cations
derived from N,N-dibenzylethylenediamine, D-glucosamine, or
ethylenediamine). The salts are prepared in a conventional manner,
e.g., by neutralizing the free base form of the compound with an
acid. It has been found that diketopiperazines suitable for use in
the present invention are present in some commercially-available
intravenous pharmaceutical compositions containing albumin,
immunoglobulin and erythropoietin. The diketopiperazines present in
these pharmaceutical preparations are formed by the heating steps
often used in the manufacture of these pharmaceutical compositions.
The heating results in cleavage and cyclization of the two
N-terminal and/or two C-terminal amino acids of the proteins to
form diketopiperazines.
[0080] Accordingly, diketopiperazines for use in the present
invention can be prepared by heating solutions of albumin,
immunoglobulin, erythropoietin and other proteins and peptides. For
example, a solution of albumin, immunoglobulin, erythropoietin or
another protein or peptide in phosphate buffer at neutral pH is
prepared. Preferably, the solution is a concentrated solution
(e.g., about 100-500 mM) to achieve protonation of the N-terminal
and/or C-terminal amino acid. The solution is heated at 60.degree.
C. for from about 2 hours to several days, preferably about 4 days,
to cause formation of the diketopiperazines. Denaturation of the
protein should, preferably, be avoided. This can be accomplished by
using shorter times and/or by adding caprylic acid or N-acetyl
tryptophan at about 0.02 M for each.
[0081] Diketopiperazines for use in the present invention can also
be prepared by contacting a solution of albumin, immunoglobulin,
erythropoietin or another protein or peptide with an enzyme that
can cleave the two N-terminal amino acids from the protein or
peptide (e.g., dipeptidyl peptidases) or an enzyme that can cleave
the two C-terminal amino acids from the protein or peptide (e.g.,
carboxypeptidases). Suitable dipeptidyl peptidases and
carboxypeptidases are available commercially from, e.g., Sigma. The
reaction should be conducted at pH 6-8, preferably in a buffer,
such as phosphate buffer, at a temperature high enough to speed the
reaction but not so high that the protein is denatured (e.g.,
37.degree. C.).
[0082] The amino acid sequences of numerous proteins and peptides
are known, and a protein or peptide with the desired N-terminal
and/or C-terminal sequence can be chosen to give the desired
diketopiperazine(s) using either method. Also, peptides with a
desired sequence can be synthesized by well known methods and
used.
[0083] The diketopiperazines can be purified from solutions
containing them, including from the commercially-available
pharmaceutical compositions comprising albumin, immunoglobulin and
erythropoietin, by well known methods, such as size-exclusion
chromatography (e.g., Centricon filtration), affinity
chromatography (e.g., using a column of beads having attached
thereto an antibody or antibodies directed to the desired
diketopiperazine(s) or an antibody or antibodies directed to the
truncated protein or peptide), anion exchange or cation exchange.
The purified diketopiperazines can be used and incorporated into
pharmaceutical compositions as described above.
[0084] A DA-DKP composition of the present invention can be
prepared from solutions containing DA-DKP, including from the
commercially-available pharmaceutical compositions comprising
albumin, such as human serum albumin, by well known methods, such
as ultrafiltration, chromatography (size-exclusion chromatography
(e.g., Centricon filtration), affinity chromatography (e.g., using
a column of beads having attached thereto an antibody or antibodies
directed to the desired diketopiperazine(s) or an antibody or
antibodies directed to the truncated protein or peptide), anion
exchange or cation exchange), sucrose gradient centrifugation,
chromatography, salt precipitation, or sonication, that will remove
some or all of the albumin in the solution. The resultant
DA-DKP-containing composition can be used and incorporated into
compositions used to coat, impregnate or cover the implantable
medical devices of the present invention.
[0085] Using ultrafiltration as a separation method, a human serum
albumin composition can be passed over an ultrafiltration membrane
having a molecular weight cut-off that retains the albumin while
the DA-DKP passes into the resulting filtrate or fraction. This
filtrate may comprise components having molecular weights less than
about 50 kDA, less than about 40 kDa, less than 30 kDa, less than
about 20 kDa, less than about 10 kDa, less than about 5 kDa, less
than about 3 kDa. Preferably, the filtrate comprises components
having molecular weights less than about 5 Da (also referred to as
"<5000 MW"). This <5000 MW fraction or filtrate contains
DA-DKP which is formed after the dipeptide aspartate-alanine is
cleaved from albumin and subsequently cyclized into the
diketopiperazine.
[0086] Implantable medical devices of the present invention are
devices which can be implanted into a subject. For example, such
devices can be a graft, a catheter, stent, prosthetic, implant
(such as a breast implant), pump, tubes, pins, rods, screws, brace,
plates or pacemaker. Stents can include but are not limited cardiac
stents and artery stents (for example for use in widening arteries
and to improve blood flow), and birth control stents (such as
Essure.RTM.). Prosthetics can include but are not limited to
artificial hip(s), artificial knee(s) or artificial ankle(s). Pumps
can include but are not limited to insulin pumps.
[0087] The implantable medical devices of the present invention can
be composed of one or more various materials. For example, the
material can be metal, steel, titanium, glass, polymers, plastics
or ceramics.
[0088] The diketopiperazines of the invention can be adhered to,
impregnated into or coated onto the surface of the implantable
medical devices. For example, an implanted medical device can be
contacted with a DKP containing solution, wherein the solution
adheres to or impregnates into or coats the implantable medical
device. The concentration of the DKP that adheres to or is coated
onto the surface of the implanted medical device can be in a range
with a lower endpoint of about 1 .mu.M, about 5 .mu.M, about 10
.mu.M, about 20 .mu.M, about 30 .mu.M, about 40 .mu.M, about 50
.mu.M, about 60 .mu.M, about 70 .mu.M, about 80 .mu.M, about 90
.mu.M, about 100 .mu.M, about 110 .mu.M, about 120 .mu.M, about 130
.mu.M, about 140 .mu.M, about 150 .mu.M, about 160 .mu.M, about 170
.mu.M, about 180 .mu.M, about 190 .mu.M, or about 200 .mu.M. In
addition, the concentration of the DKP that adheres to or is coated
onto the surface of the implanted medical device can be in a range
with an upper endpoint of about 500 .mu.M, about 475 .mu.M, about
450 .mu.M, about 425 .mu.M, about 400 .mu.M, about 375 .mu.M, about
350 .mu.M, about 325 .mu.M, about 320 .mu.M, about 310 .mu.M, about
300 .mu.M, about 290 .mu.M, about 280 .mu.M, about 270 .mu.M, about
260 .mu.M, about 250 .mu.M, about 240 .mu.M, about 230 .mu.M, about
220 .mu.M, or about 210 .mu.M.
[0089] In a further embodiment of the present invention, the
concentration of the DKP on a surface of a medical device that is
exposed to the body when implanted can be in a range with a lower
endpoint of about 1 ng/cm.sup.2, about 5 ng/cm.sup.2, about 10
ng/cm.sup.2, about 15 ng/cm.sup.2, about 20 ng/cm.sup.2, about 25
ng/cm.sup.2, about 30 ng/cm.sup.2, about 35 ng/cm.sup.2, about 40
ng/cm.sup.2, about 45 ng/cm.sup.2, about 50 ng/cm.sup.2, about 55
ng/cm.sup.2, about 60 ng/cm.sup.2, about 65 ng/cm.sup.2, about 70
ng/cm.sup.2, about 75 ng/cm.sup.2, about 80 ng/cm.sup.2, about 85
ng/cm.sup.2, about 90 ng/cm.sup.2, about 95 ng/cm.sup.2, or about
100 ng/cm.sup.2. Preferably, the concentration of the DKP on a
surface of a medical device that is exposed to the body when
implanted is about 50 ng/cm.sup.2. In addition, the concentration
of the DKP on a surface of a medical device that is exposed to the
body when implanted can be in a range with an upper endpoint of
about 200 ng/cm.sup.2, about 195 ng/cm.sup.2, about 190
ng/cm.sup.2, about 185 ng/cm.sup.2, about 180 ng/cm.sup.2, about
175 ng/cm.sup.2, about 170 ng/cm.sup.2, about 165 ng/cm.sup.2,
about 160 ng/cm.sup.2, about 155 ng/cm.sup.2, about 150
ng/cm.sup.2, about 145 ng/cm.sup.2, about 140 ng/cm.sup.2, about
135 ng/cm.sup.2, about 130 ng/cm.sup.2, or about 125
ng/cm.sup.2.
[0090] Subjects of the present invention can be a mammal, such as a
rabbit, goat, dog, cat, horse or human. Preferably, the subject is
a human.
[0091] As used herein, "a" or "an" means one or more.
[0092] As used herein, "comprises" and "comprising" include within
their scope all narrower terms, such as "consisting essentially of"
and "consisting of" as alternative embodiments of the present
invention characterized herein by "comprises" or "comprising". In
regard to use of "consisting essentially of", this phrase limits
the scope of a claim to the specified steps and materials and those
that do not materially affect the basic and novel characteristics
of the invention disclosed herein.
[0093] Additional objects, advantages and novel features of the
present invention will become apparent to those skilled in the art
by consideration of the following non-limiting examples. The
following experimental results are provided for purposes of
illustration and are not intended to limit the scope of the
invention.
EXAMPLES
Example 1
[0094] This example shows the results of an analysis of biofilms on
extracted orthopedic devices to determine if the presence of a
diketopiperazine is correlated with bacterial colonization.
Bacteria use small molecular weight N-acylhomoserine lactones and
diketopiperazines to initiate biofilm formation and regulate colony
growth. An Aspartate, Alanine-Diketopiperazine (DA-DKP) formed by
the cleavage and cyclization of the N-terminal amino acids of human
serum albumin has previously been demonstrated to be
immunomodulatory for memory but not naive human T lymphocytes.
[0095] Methods: This study was an institutional review board (IRB)
approved study. Twenty-two patients undergoing hardware removal
were enrolled. The removed orthopedic devices were stripped of
surface biofilm using methanol/ammonium formate. The .ltoreq.3 kD
MW material was collected and diketopiperazine levels analyzed
using anion exchange high pressure liquid chromatography coupled to
negative electrospray ionization mass spectrometry.
[0096] Results: The thirty-three patients ranged in age from 6 to
91 years, with a mean of 54. There were fifteen males and eighteen
females. Ten devices were reported by the clinical laboratory to be
culture positive. In five cases the main organism was
Staphylococcus. In one of the three cases Bacteroides and
Streptococcus species were also isolated. In all thirty-three cases
detectable amounts of DA-DKP were identified with a mean level of
120 ng/ml. Higher amounts of DA-DKP (9.75-235 ng/ml) were detected
in the culture positive devices versus the culture negative
(1.78-34.7 ng/ml). In addition, one device removed from a case with
osteomyelitis had a DA-DKP content of 3,063 ng/ml. (see Table
1).
[0097] Conclusion: DA-DKP is an important immune modulator in
biofilm formation on orthopedic implants. Its presence in biofilms
found on extracted orthopedic devices suggests innate physiologic
mechanisms conferring tolerance to the implanted device possibly
correlated to the presence of inflammation/rejection reactions.
TABLE-US-00001 TABLE 1 Study Group DA-DKP amounts Sex of subject
Concentration Age of F = female Culture of DA-DKP subject M = male
Device Removed Results ng/ml 73 F Pin (hip) None 4.63 66 F Rod +
Screws (hip) Negative 19.7 89 F Rod + Screws (hip) Staph 87.0
(positive) 74 M Staple (knee) None 3.03 67 F Septic hip prosth.
Staph aureus 84.8 44 M Plate (clavicle) None 2.40 45 F Rod + Screws
(spine) Negative 30.9 87 F Screws (hip arthritis) None 19.9 33 M
Rod + Screws (knee) None 6.03 45 M Rod + Screws (ulna) None 9.05 31
F Rod + Screws (ankle) Bacteroides + 53.7 Strep 52 F Brace + Screws
None 9.02 (tibia) 58 F Screws (femur) Staph 28.4 (positive) 74 M
Brace (clavicle) Negative 7.02 6 M Plate + Screws None 22.4 60 F
Rod (osteomyelitis Negative 3,063 tibia) 58 F Hip Prosth. Negative
23.0 66 F Rod (femoral) Staph + Strep 235 45 M Rod + Screws (tibia)
Gram + 12.9 55 M Nails (femoral) Gram + 70.4 54 F Ankle Prosth.
Negative 34.7 24 M Screws (tibia) Gram + 9.75 91 F Screws (hip)
None 3.91 44 M Plate + Screws None 14.2 (ankle) 46 F Screws (ankle)
None 9.32 24 M Screws (femur) Staph aureus 53.1 68 F Knee Prosth.
Negative 1.78 56 F Screws (knee) None 3.41 41 F Brace + Screws
Yeast 20.4 (ankle) 61 M Screws (femur) Negative 4.21 34 M Plate +
Screws Negative 9.78 (radius) 77 M Plate + Rod (hip) None 2.13 28 M
Plates + Screw None 12.7 (ankle)
Example 2
[0098] Isolation and characterization of peptides and proteins from
endotracheal tubes. The results of this example further demonstrate
that DKPs form on implantable medical devices such as endotracheal
tubes, when they are implanted within a subject. The presence of
the DKPs on these tubes helps the subject to confer tolerance to
the tubes. This again demonstrates the unique finding of coating
implantable medical devices, such as endotracheal tubes, with DPKs
prior to implantation so as to increase the subject's immune
tolerance and/or to decrease a subject's inflammatory response to
the tube.
[0099] Endotracheal tubes discarded from mechanically ventilated
trauma patients are collected into sterile biohazard pouches and
transported immediately to the Trauma Research Lab. As controls for
the absence of biofilms, discarded endotracheal tubes form surgical
patients that were only used for a few hours during surgery were
used.
Method
[0100] Biofilm and/or mucus is stripped from the proximal ends of
endotracheal tubes by placing in a sterile centrifuge tube
containing 1-2 ml of chromatography each analysis buffer consisting
of methanol 60% plus 50 mM ammonium formate 40% with extensive
washing using a pipette and agitation on a vortexer. After the
biofilm is stripped from the endotracheal tube, the sediment is
pelleted by centrifugation and frozen for later analysis of
bacterial content. The biofilm supernatant is collected for
analysis of protein and large molecular width peptides. An aliquot
of the biofilm supernatant is placed in an ultrafiltration spin
column (Vivaspin 500, 3,000 MWCO, Sartorius, Hannover, Germany) for
centrifugation at 15,000.times.g. The filtrate is collected for
analysis of <3 kD molecular weight peptides.
[0101] Supernatants containing higher molecular weight material are
analyzed by high performance liquid chromatography (HPLC, Waters,
Milford, Mass., USA) coupled to positive electrospray ionization
time of flight mass spectrometry (+ESI-TOF MS, Micromass, UK). Each
supernatant is diluted 1:10 with dH.sub.2O. 10 .mu.L of each sample
is injected onto a YMC-Pack Protein-PR, 150 mm.times.4.6 mm, 5 u,
HPLC column heated at 50.degree. C. (Waters, Milford, Mass., USA)
using a 20-minute linear gradient method used water/0.1%
trifluoroacetic acid (A) and acetonitrile/0.1% TFE (B). The output
of the HPLC is split 1:20 (v:v) and injected into the mass
spectrometer with a scan range of 500 to 3500 m.z, cone voltage of
30 eV, source temperature of 100.degree. C., and gas temperature of
250.degree. C. Albumin (a molecular standard) elutes at 8.15
minutes and is visualized as a charge envelope from 1100 to 2500
m/z representing +44 to +26 charges. The spectrum is the
deconvolved to the uncharged parent mass using MaxEnt 1 (Micromass,
UK). The parent mass spectrum is then integrated and relative
proportions of each species were calculated.
[0102] 50 .mu.l of each of the <3000-Da filtrate fractions of
biofilm supernatant is injected into high performance liquid
chromatography (HPLC, 2795 system, Waters, Mass.) coupled to a mass
spectrometer (LCT-TOF, Micromass, UK), and quantified using a
storage anion exchange column (Supelcosil, SAX1 250 mm.times.4.6
mm, Supelco) and a 70:30 v/v methanol/water with 25 mM ammonium
acetate (Sigma Aldrich, St. Lois, Mo.) as the mobile phase in an
isocratic mode at 1 ml/min. The output of the HPLC is split 1:20
(v/v) and injected into the mass spectrometer using negative
electrospray ionization (-ESI MS) with a scan ranges of 80-1000
m/z, cone voltage of 30 eV, source temperature of 100.degree. C.
and a gas temperature of 250.degree. C. DA-DKP, as a molecular
standard, is measure by monitoring the mass 185 in time which
corresponds to DA-DKP minus a single proton (-H+). DA-DKP elutes at
5.8 mins and is quantified by integrating the area under the curve.
The area was compared with a standard curve derived from synthetic
DA-DKP standard (DMI Synthesis, Newport, Wales) of known
concentrations (5000 ng/ml, 1000 ng/ml, 200 ng/ml, 40 ng/ml, 8
ng/ml). The calibration curve was found to be very linear in this
range within R2 of 0.99998.
[0103] The concentration of DKP on over 100 endotracheal tubes as
detected by the method described above is presented in Table 2. The
DKP concentration ([DKP]) provided on Table 2 has already been
adjusted per volume added to dissolve biofilm. The following are
indicated on Table 2:
[0104] ID #: Subject identification number
[0105] Sex: Subject's sex either male (M) or female (F)
[0106] Age: Age in years of the subject
[0107] [DKP] ng/ml: The DKP concentration in ng/ml already adjusted
per volume added to dissolve biofilm
[0108] Bacteria identified: Type of bacteria detected on
endotracheal tube
[0109] Vent Days: The number of days the endotracheal tube was
implanted with the subject
[0110] Protein identified: The proteins that were determined on the
endotracheal tube
[0111] AIS: Abbreviated injury score/scale, with a score of 1 being
a minor injury, 2=moderate, 3=serious, 4=severe, 5=critical,
6=maximum, 9=not further specified.
[0112] ISS: Injury severity score, assesses trauma severity and
correlates with mortality, morbidity and hospitalization time after
trauma.
[0113] GCS: Glascow coma score/scale-neurological scale to help
assess the status of the central nervous system and used acutely to
grade the severity of a subject's trauma and mental function.
[0114] GOS: Glascow outcome score/scale (R=rehabilitation; L=long
term acute care; 1=dead; 5=good recovery)--a 5-point score given to
victims of traumatic brain injury at some point in their
recovery.
TABLE-US-00002 TABLE 2 DKP Concentration on Endotracheal Tubes ID
[DKP] Vent Proteins # Sex Age ng/ml Bacteria Days ID AIS ISS GCS
GOS 18 M 25 150.06 8 PRP1, 2 38 15 5 HIG2, AT5G1 24 M 47 607.73
S.pneumon 11 Defensin 3 20 15 R MRSA 1, 2, 3 LL-37, Lysozyme C 25 M
90 <20 3 Defensin 3 14 6 1 1, 2, 3 LL-37 28 F 22 <20 1
Defensin 3 21 7 R 1, 2, 3 LL-37 30 F 55 1,071.55 Enterob. 4 2 17 3
R Sakazakii 30-2 F 55 <20 Enterob. 4 2 17 3 R Sakazakii 31 M 60
1,402.85 9 S10A8 3 10 15 5 34 M 20 <20 H. 30 5 45 3 R influenza
35 F 71 116.29 7 4 30 6 L 36 M 46 1,580.08 Pneumo- 11 3 34 9 R
thorax 36-2 M 46 <20 Pneumo- 11 3 34 9 R thorax 39 M 58 <20 5
4 29 15 L 41 M 35 72.53 K. oxytoca 20 3 43 3 L S. aureus 44 M 35
<20 8 4 25 4 5 45 M 17 34.09 S. 3 1 21 3 5 marcesens 50 M 76
362.29 1 3 22 52 M 34 159.56 <1 2 17 12 R 53 M 21 156.85 <1 3
14 6 1 58 M 25 <20 36 5 29 15 L 60 M 15 124.82 1 1 41 5 70 M 28
104.63 4 3 26 3 R 79 M 73 33.48 K. oxytoca 8 4 L 87 M 49 79.92 90 F
56 103.79 96 F 49 637.7 97 M 30 1,752.89 98 F 24 952.71 102 M 32
1,311.48 106 F 80 1,083.15 108 F 59 2,648.23 109 M 20 629.67 114 M
76 379.51 121 F 84 2,325.85 123 M 15 8,009.14 129 M 27 500.95 131 F
65 16,194.74 132 M 32 282.84 133 F 48 480.59 138 M 49 137.37 140 F
81 567.2 143 M 28 426.93 145 M 68 1,655.73 149 M 21 58.68 149-2 M
21 749.03 150 M 31 111.68 151 M 50 389.23 152 M 25 134.8 153 F 51
272.45 156 M 29 25.45 157 F 18 634.13 158 M 51 2,183.93 161 M 77
386.28 161-2 M 77 691.98 162 M 18 364.63 163 M 63 107.4 164 F 15
69.15 166 F 18 856.74 169 F 30 <20 170 M 73 368.9 178 M 53 48.55
179 M 25 <20 182 M 67 1059.7 182-2 M 67 255.73 183 M 36 279.76
184 M 37 <20 190 M 34 624.44 191 M 29 2,207.58 191-2 M 29 798.37
192 M 66 <20 195 M 28 2,608.52 196 F 23 <20 198 M 35 404.87
200 M 33 816.38 202 M 67 <20 203 M 50 112.64 204 1,290.96 205
2,108.70 206 656.8 208 <20 209 276.96 210 76.19 211 76.98 212
<20 212-2 505.63 213 218.41 215 503.16 215-2 322.54 216 163.25
219 12,019.74 221 177.19 222 155.14 225 247.88 228 190.86 229 92.4
231 243.68 233 72.03 234 22.96 235 356.11 235-2 68.11 237 509.7 239
1,620.48 Surg 2,648.55 <1 001 Surg 500.22 <1 002 Surg 56.74
<1 003 Surg 505.67 <1 004 Surg <20 <1 005 Surg F 83
89.23 92 Surg F 83 67.87 92-2 R 2235.63 111 R 1216.05 113
[0115] While various embodiments of the present invention have been
described in detail, it is apparent that modifications and
adaptations of those embodiments will occur to those skilled in the
art. It is to be expressly understood, however, that such
modifications and adaptations are within the scope of the present
invention, as set forth in the following exemplary claims.
* * * * *