U.S. patent application number 11/677576 was filed with the patent office on 2007-08-23 for high-yield activation of polymer surfaces for covalent attachment of molecules.
This patent application is currently assigned to THE TRUSTEES OF PRINCETON UNIVERSITY. Invention is credited to T. Joseph Dennes, Jeffrey Schwartz.
Application Number | 20070196663 11/677576 |
Document ID | / |
Family ID | 38438106 |
Filed Date | 2007-08-23 |
United States Patent
Application |
20070196663 |
Kind Code |
A1 |
Schwartz; Jeffrey ; et
al. |
August 23, 2007 |
HIGH-YIELD ACTIVATION OF POLYMER SURFACES FOR COVALENT ATTACHMENT
OF MOLECULES
Abstract
Polymer surfaces coated with organometallic layers, wherein the
organometallic layers and polymer surfaces have functional groups
that react to bond the organometallic layer to the polymer surface
with organometallic functional groups remaining unreacted for the
subsequent covalent attachment of organic overlayers. Coating
methods and coated articles are also disclosed.
Inventors: |
Schwartz; Jeffrey;
(Princeton, NJ) ; Dennes; T. Joseph; (Cranbury,
NJ) |
Correspondence
Address: |
SYNNESTVEDT LECHNER & WOODBRIDGE LLP
P O BOX 592
112 NASSAU STREET
PRINCETON
NJ
08542-0592
US
|
Assignee: |
THE TRUSTEES OF PRINCETON
UNIVERSITY
New South Building, 4th Floor P.O. Box 36
Princeton
NJ
08544
|
Family ID: |
38438106 |
Appl. No.: |
11/677576 |
Filed: |
February 21, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60775127 |
Feb 21, 2006 |
|
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|
60804633 |
Jun 13, 2006 |
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Current U.S.
Class: |
428/411.1 ;
427/2.1; 428/420 |
Current CPC
Class: |
A61L 27/14 20130101;
C08H 1/06 20130101; C08J 7/0423 20200101; A61L 31/082 20130101;
A61L 27/30 20130101; A61L 27/50 20130101; C08J 2379/02 20130101;
Y10T 428/31536 20150401; A61L 27/38 20130101; A61L 27/28 20130101;
A61P 43/00 20180101; C09J 189/04 20130101; Y10T 428/31504 20150401;
A61P 35/00 20180101; A61L 31/005 20130101 |
Class at
Publication: |
428/411.1 ;
427/002.1; 428/420 |
International
Class: |
B32B 27/00 20060101
B32B027/00; A61L 33/00 20060101 A61L033/00 |
Claims
1. A coated substrate comprising a polymer surface and an
organometallic layer formed thereon, wherein the organometallic
layer and polymer surface have functional groups that react to bond
the organometallic layer to the polymer surface.
2. The coated substrate of claim 1, wherein said organometallic
layer comprises transition metal atoms selected from the group
consisting of atoms of Group 4, Group 5 and Group 6 of the Periodic
Chart.
3. The coated substrate of claim 1, wherein said organometallic
layer comprises alkoxide or dialkylamide groups at least a portion
of which react with said functional groups of said polymer
surface.
4. The coated substrate of claim 1, wherein said organometallic
layer has unreacted functional groups and said coated substrate
further comprises an organic overlayer coated on said
organometallic layer, said overlayer comprising a compound,
polypeptide, oligomer or polymer having functional groups that
react with said unreacted organometallic layer functional groups to
bond said overlayer to said organometallic layer.
5. The coated substrate of claim 4, wherein said overlayer
comprises an organophosphorus, organocarboxylic acid or
organocarboxylic acid ester compound covalently bonded to said
organometallic layer.
6. The coated substrate of claim 5, wherein the organo group of the
organophosphorus, organocarboxylic acid or organocarboxylic acid
ester overlayer compound is a saturated or unsaturated, substituted
or unsubstituted alkyl group.
7. The coated substrate of claim 6, wherein said aalkyl group is
substituted in the omega position and the omega-substituents of the
omega-substituted organophosphorus, organocarboxylic acid or
organocaroxylic acid ester overlayer compounds are selected from
the group consisting of carboxylate, carbamate, hydroxyl, keto,
ether, oxy, carbonate, amino, amide and thiol.
8. The coated substrate of claim 5, wherein said overlayer
comprises an organo-phosphorus compound selected from the group
consisting of phosphoric acids, phosphonic acids and phosphinic
acids.
9. The coated substrate of claim 8, wherein said organophosphorus
overlayer comprises an organophosphoric acid compound or mixture of
compounds of the structure: (RO).sub.x--P(O)--(OR').sub.y wherein x
is 1 or 2, y is 1 or 2 and x+y=3; R is a radical having a total of
1-30 carbons; where R' is H, a metal or lower alkyl having 1-4
carbons; and, for at least a portion of the organo-phosphorus
compounds in the overlayer, R' is H.
10. The coated substrate of claim 8, wherein said organophosphorus
overlayer comprises an organophosphonic acid compound or mixture of
compounds of the structure: ##STR7## wherein x is 0 or 1, y is 1, z
is 1 or 2 and x+y+z is 3; R and R'' are each independently radicals
having a total of 1 to 30 carbons; R' is H, a metal or lower alkyl
having 1-4 carbons, and, for at least a portion of the
organophosphorus compounds in the overlayer, R' is H.
11. The coated substrate of claim 8, wherein said organophosphorus
overlayer comprises an organophosphonic acid compound or mixture of
compounds of the structure: ##STR8## wherein x is 0, 1 or 2, y is
0, 1 or 2, z is 1 and x+y+z is 3; R and R'' are each independently
radicals having a total of 1 to 30 carbons; R' is H, a metal or
lower alkyl and, for at least a portion of the organophosphorus
compounds in the overlayer, R' is H.
12. The coated substrate of claim 5, wherein said overlayer
comprises an organo-phosphorus or organocarboxylic acid compound
with an organo group containing a C.sub.6 to C.sub.18 hydrocarbon
or substituted hydrocarbon group.
13. The coated substrate of claim 1, wherein the substrate is a
molded polymer article.
14. The coated substrate of claim 1, wherein the polymer is in the
form of a coating.
15. The coated substrate of claim 14, wherein the polymer is a
coating on an article made from another material, such as glass,
silicon dioxide, metal, or another polymer.
16. The coated substrate of claim 1, wherein the polymer surface
has surface functional groups selected from the group consisting of
amide, imide, urethane, urea, amine, epoxy, hydroxyl, oxy, keto,
acidic C--H, phenol, carboxylic acid, carboxylic acid ester,
carboxylic acid anhydride, sulfonic acid, and thiol groups.
17. The coated substrate of 1, wherein said polymer surface is a
polymer selected from the group consisting of polyamides,
polyimides, polyurethanes, polyureas, polyamines, polyepoxides,
polyesters containing unreacted hydroxyl or carboxylic acid groups,
polysulf-onamides and polysulfides containing unreacted thiol
groups.
18. The coated substrate of claim 4, wherein said overlayer
comprises a biologically or pharmaceutically active compound
covalently bonded to the organometallic coating layer.
19. The coated substrate of claim 6, further comprising a second
overlayer of a biologically or pharmaceutically active compound
covalently bonded to said functional groups of said
organophorphorus, organocarboxylic acid or organocarboxylic acid
ester compound overlayer.
20. The coated substrate of claim 4, wherein said overlayer
comprises a biologically active ligand compound selected from cell
attachment mediators or a substance that enhances or excludes
particular varieties of cellular or tissue ingrowth.
21. The coated substrate of claim 20, wherein said biologically
active ligand compound is selected from the group consisting of
osteo-inductive substances and substances that induce cellular
growth and proliferation and integrin cell attachment
mediators.
22. The coated substrate of claim 21, wherein said biologically
active ligand compound is selected from the group consisting of
bone morphogenic proteins (BMP), epidermal growth factor (EGF),
fibroblast growth factor (FGF), platelet-derived growth factor
(PDGF), insulin-like growth factor (IGF-I and II), TGF- and
vascular endothelial growth factor (VEGF).
23. The coated substrate of claim 4, wherein said overlayer
comprises a pharmaceutically active compound selected from the
group consisting of anti-neoplastic and anti-proliferative
agents.
24. The coated substrate of claim 4, wherein said overlayer
comprises a active agent selected from the group consisting of
acyclovir, cephradine, malphalen, tamoxifen, raloxifene,
daunomycin, adriamycin, plumbagin, chlorambucil, ephedrine,
atropine, quinine, digoxin, quinidine, biologically active
peptides, chlorin e.sub.6, cephalothin, proline, proline analogues,
penicillin V, aspirin, ibuprofen, steroids and nicotinic acid.
25. A polymer scaffold for tissue engineering comprising the coated
substrate of claim 1.
26. A polymer scaffold for tissue engineering comprising the coated
substrate of claim 4, wherein said overlayer comprises a
biologically active ligand for cellular or tissue ingrowth.
27. The polymer scaffold of claim 26, wherein said scaffold is
adapted for the re-generation of nervous, musculo-skeletal,
cartilaginous, tendenous, hepatic, pancreatic, ocular,
integumentary, arterio-venous or urinary tissues or tissues forming
solid or hollow organs.
28. The polymer scaffold of claim 26, containing cells selected
from the group consisting of cells of the muscular and skeletal
systems, parenchymal cells, cells of intestinal origin, exocrine
cells, bile duct cells, parathyroid cells, thyroid cells, cells of
the adrenal-hypothalmic-pituitary axis, heart muscle cells, kidney
epithelial cells, kidney tubular cells, kidney basement membrane
cells, nerve cells, blood vessel cells, cells forming bone and
cartilage, smooth muscle cells, skeletal muscle cells, ocular
cells, integumentary cells, keratinocytes, skin cells and
endothelial cells.
29. The polymer scaffold of claim 28, wherein said cells are
selected from the group consisting of chondrocytes, fibroblasts,
osteocytes, osteoblasts, hepatocytes and pancreatic cells.
30. The polymer scaffold of claim 26, containing cells selected
from the group consisting of cells obtained from donors, embryonic
stem cells, non-embryonic stem cells, cells from established cell
culture lines, cells before genetic engineering and cells after
genetic engineering.
31. The polymer scaffold of claim 30, wherein said cells from
established cell culture lines comprise cells from embryonic stem
cell culture lines and cells from non-embryonic stem cell culture
lines.
32. A method of regulating cellular attachment, migration and
proliferation on a polymeric substrate, characterized by contacting
living cells, tissues or biological fluids containing living cells
with the polymer scaffold of claim 26.
33. An implantable medical device characterized by one or more
surfaces comprising the coated substrate of claim 1.
34. The implantable medical device of claim 33, characterized in
that it is a vascular graft, stent, bone plate, suture, implantable
sensor, barrier for surgical adhesion prevention or implantable
drug delivery device.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims priority benefit under 35
U.S.C. .sctn.119(e) of U.S. Provisional Patent Application Nos.
60/775,127 filed Feb. 21, 2006 and 60/804,633 filed Jun. 13, 2006.
The disclosures of both applications are incorporated herein by
reference.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to covalently binding organic
materials to the surfaces of polymer substrates by functionalizing
the surfaces with linker moieties containing transition metal
complexes. In particular, the present invention relates to
modifying polymer surfaces with organometallic compounds that have
functional groups that react with functional groups of the polymer
surface. The modified polymer surface can be further reacted with a
compound, polymer or oligomer that contains functional groups that
are reactive with functional groups of the organometallic compound
that remain after reaction with the polymer surface. The present
invention further relates to functionalizing polymer surfaces to
support cell growth and the attachment of biologically active
molecules and other compounds of interest.
[0003] Bioactive polymeric scaffolds are of increasing importance
for use in tissue regeneration in a variety of clinical
applications, and a scaffold that supports cell growth is a
critical first step in such regeneration. Surface wetting
properties of many polymers used as bioscaffolds are not conducive
to biointegration, but incorporation of surface functional groups
can effect substantial changes in a polymer's wettability while
generating reactive sites suitable for attachment of peptides and
other biomolecules.
[0004] Adjustment of the surface properties of polymers such as
those from which preformed polymeric therapeutic devices are
formed, has proven problematic because those polymers most often
used as biomaterials are resistant to specific surface treatments.
To circumvent this problem, polymer scaffold materials have been
prepared by blending, copolymerization, or physical treatment, but
these methods can result in alteration of the bulk properties of
the polymer. Furthermore, these methods or attempts to surface
modify pre-cast polymers using standard methods of organic
synthesis, result only in low surface coverage by peptides that do
not approach those that can be achieved on metallic substrates.
[0005] Surface modification has also proven problematic in other
areas of polymer and polymer surface customization. There is a need
to modify polymer surfaces so as to change the surface
characteristics of the polymer surface without changing the bulk
properties of the polymer. More particularly, there remains a need
for polymer surfaces that support cell growth as well as the
attachment of biologically active molecules and other compounds of
interest.
SUMMARY OF THE INVENTION
[0006] The present invention relates to a coated substrate having a
polymer surface and an organometallic coating layer formed thereon.
The organometallic coating and polymer surface have co-reactive
functional groups that are reacted to bond the coating to the
surface. Preferably, the organometallic coating has remaining
unreacted functional groups that are reacted with co-reactive
functional groups of a subsequently applied compound, oligomer or
polymer.
[0007] In another embodiment, the present invention relates to a
method of coating a polymer surface, which includes coating an
organometallic compound on the polymer surface, wherein the polymer
surface and the organometallic coating have co-reactive functional
groups; and reacting the functional groups of the organometallic
compound with the co-reactive functional groups of the polymer
surface to form an organometallic coating on the polymer surface.
Preferably, a compound, oligomer or polymer containing functional
groups that react with the remaining unreacted functional groups of
the organometallic compound is applied to the organometallic layer
and the functional groups of the compound, oligomer or polymer are
reacted with the remaining functional groups in the organometallic
layer to form an organic coating on the organometallic coating.
[0008] In one embodiment, the present invention provides a novel
approach to polymer surface modification that enables high surface
density derivatization of polymers with reactive surface moieties
containing acidic covalent bonds. The polymers need not contain
pendant functional groups with acidic covalent bonds. For example,
polymers with exposed amide functionality contain acidic N--H
bonds, which can serve as sites for chemical derivatization when
appropriately activated. In particular, coordination of the
carbonyl group to an appropriate metallic center further acidifies
the N--H bonds and facilitates derivatization. In addition, acidic
C--H bonds on polymer surfaces can also serve as sites for chemical
derivatization when appropriately activated. In particular,
coordination of the carbonyl group to an appropriate metallic
center further acidifies the C--H bonds and facilitates
derivatization.
[0009] Therefore, according to one aspect of the present invention,
a coated substrate is provided having a polymer surface with
exposed reactive functional groups containing acidic covalent bonds
and an organometallic coating layer formed thereon and covalently
bonded thereto, wherein the organometallic coating layer contains
transition metal atoms selected from atoms of Group 4, Group 5 and
Group 6 of the Periodic Chart that have been covalently bonded to
the polymer surface by reaction of a polyalkoxide or
polydialkylamide of the transition metal with the reactive
functional groups exposed on the polymer surface. Zirconium and
titanium are two examples of such transition metals. Examples of
reactive functional groups with acidic covalent bonds include, but
are not limited to, hydroxyl groups, phenol groups, amide N--H
groups, amino groups, imide groups, urethane groups, urea groups,
thiol groups, carboxylic acid groups, carboxylic acid ester groups,
carboxylic acid amide groups, sulfonic acid groups, acidic C--H
groups, and the like. Each of these groups will covalently bond the
organometallic coating layers.
[0010] The organometallic layer can then be further bonded to
organic groups or ligands of interest that are reactive with the
organometallic layer, thereby attaching covalently the organic
ligands of interest to the polymer surface. Therefore, according to
another aspect of the present invention, a coated substrate is
provided having a polymer surface with an organometallic coating
layer formed thereon and covalently bonded thereto, wherein the
organometallic coating layer contains transition metal atoms
selected from atoms of Group 4, Group 5 and Group 6 of the Periodic
Chart which have been covalently bonded to the polymer surface by
reaction of a polyalkoxide or polydialkylamide of the transition
metal with the reactive functional groups on the polymer surface,
wherein each transition metal atom additionally has covalently
bonded to it one or more organic ligands.
[0011] The present invention provides coated substrates in which up
to 40%, up to 60%, up to 75% or up to 100% of the polymer surface
has organic groups or ligands bonded thereto. This is accomplished
without changing the bulk material properties of the substrate. The
percentage of the polymer surface with organic ligands bonded
thereto depends on relationship between the size of the organic
ligand and the size of the metal complex used to bond the organic
ligand to the polymer and is readily apparent to one of ordinary
skill in the art guided by the present specification.
[0012] The present invention thus provides a novel type of
interface that enables strong adhesion between a polymer surface
and an organic coating. The coating modifies the surface of the
polymer making it more hydrophilic or hydrophobic as may be
desired. The modification can make the polymer surface more or less
receptive to subsequently applied coatings, improving or minimizing
the adhesion of the polymer to the subsequently applied coating.
The polymer surface can be a polymer coating on an article made
from another material, such as glass, silicon dioxide, metal, or
another polymer. The polymer surface can also be the surface of a
pre-cast polymer article. Examples of suitable polymers with
reactive functional groups include polyamides, such as nylon, silk,
and collagen, polyacrylamides, polyimides, polyurethanes,
polyureas, polysulfonamides, polyesters, polysaccharides, such as
haluronic acid, methylcellulose and proteoglycans, and the like,
and copolymers of any of these polymers.
[0013] According to one embodiment of this aspect of the invention,
the polymer is a biocompatible polymer and the polymer-coated or
polymer-cast article is a medical implant or biological scaffold or
porous matrix. The organic ligand can then be a protein, peptide,
peptide mimetic, small molecule ligand for a cell surface receptor,
or other biologically or pharmaceutically active compound having
utility as a coating on a medical implant. The organic ligand can
thus promote or prevent cell growth or proliferation, promote or
discourage cell adhesion, prevent infection, or prevent or promote
blood clotting or adhesion.
[0014] According to another embodiment of this aspect of the
invention, the polymer is a fabric formed from a woven or non-woven
fiber. The fiber can be a natural fiber with exposed functional
groups, such as silk, wool, cotton, linen, collagen and the like.
The fiber can also be a synthetic fiber with exposed functional
groups, such as nylon.
[0015] The present invention also provides a method by which
organic ligands or groups may be covalently bonded to polymer
surfaces with reactive functional groups using an organo-metallic
interface. Therefore, according to another aspect of the present
invention, there is provided a method of forming an organic layer
on a polymer surface with reactive functional groups, which method
includes the steps of:
[0016] providing a substrate having a polymer surface covalently
bonded to an organometallic surface layer of dialkylamides or
alkoxides of transition metals selected from Group 4, Group 5 or
Group 6 of the Periodic Chart, wherein the dialkylamides or
alkoxides are bonded at the transition metal atoms to the polymer
surface; and
[0017] reacting the transition metal dialkylamide or alkoxide layer
with an organic overlayer comprising a compound, oligomer or
polymer capable of reacting with unreacted transition metal
dialkylamide or alkoxide groups to covalently bond the organic
compound, oligomer or polymer to the transition metals.
[0018] The polymer surface can be provided with the organometallic
surface layer by reacting a polymer substrate having reactive
functional groups with acidic covalent surface bonds with a
polydialkylamide or polyalkoxide of the Group 4, Group 5 or Group
6transition metal having two or more dialkylamide or alkoxide
groups, so that an organometallic surface layer is formed,
covalently bonded to the polymer surface, and having at least one
unreacted dialkylamide or alkoxide group.
[0019] The method of the present invention thus provides high yield
coatings on polymer surfaces with the adhesion properties of
physical deposition methods under mild reaction conditions. In
particular, the coatings of the present invention may be formed at
ambient temperatures.
[0020] In addition to the coatings of the present invention and the
method by which they are formed, the present invention also
provides coated implantable medical devices, methods for improving
cellular growth and attachment, tissue in-growth and adhesion to
tissue for implantable medical devices using the coatings of the
invention and the inventive coating methods, and methods for
implanting medical devices by first coating them according to the
present invention
[0021] The invention can also be used to make the polymer surface
electrically conductive, semi-conductive or electrically insulating
making it useful in organic thin film transistors, light emitting
devices and electrolytic capacitors.
[0022] Other features of the present invention will be pointed out
in the following description that discloses, by way of example, the
principles of the invention and the best methods which have been
presently contemplated for carrying them out.
BRIEF DESCRIPTION OF THE DRAWINGS
[0023] FIG. 1 depicts the reaction of N-hexylacetamide and
zirconium tetra(tert-butoxide);
[0024] FIG. 2 depicts the reaction of nylon-Zr-amide complex with a
phosphonic acid and RGDC coupling;
[0025] FIG. 3 depicts the transesterification reaction of
nylon-Zr-amide complex and RGDC coupling; and
[0026] FIGS. 4a-4c depict fibroblast cell growth on surface
embodiments of the invention according to one embodiment of a
method of the invention.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0027] The coated substrates are formed by reacting a polymer
surface having functional groups with acidic covalent bonds that
are reactive with other functional groups, specifically transition
metal polydialkylamides and polyalkoxides. For purposes of the
present invention, "reactive groups" on a polymer surface are
defined as functional group with acidic covalent bonds. Examples of
suitable polymer surface reactive functional groups include those
having a reactive N--H bond such as amine, amide, imide, urethane
and urea groups. Examples of other reactive functional groups
include hydroxyl, oxy, ether, thiol, carbonyl including keto,
ester, free acid and acid anhydride, sulfonic acid and acidic--CH
groups.
[0028] The polymer can be in the form of a molded article, a rigid
or flexible film or a coating. Examples of suitable polymers
containing these groups are polyamines such as polyoxy-alkylene
polyamines, polyethers such as polyethylene glycol, polyketones
such as PEEK, polyamides such as nylon, polyacrylamides,
polyimides, polyesters and polyurethanes such as the reaction
product of polymeric polyols with polyisocyanates such as
techoflex. The preferred functional groups are ##STR1## groups such
as those associated with polyamides. Suitable polyamides include
nylons such as Nylon 6. Nylon 4/6, Nylon 6/6, Nylon 6/9, Nylon
6/10, Nylon 6/12, Nylon 12, Nylon 6/66, and the like.
[0029] The reactive functional groups containing acidic covalent
bonds may be either pendant to or between monomeric repeating units
of the polymer, a portion of which are exposed at the polymer
surface.
[0030] The organometallic compound used in the practice of the
invention is preferably derived from a metal or a metalloid such as
selected from Group 3 of the Periodic Chart or a transition metal
selected from Group 4, 5 and 6 of the Periodic Chart. Preferred
metals are aluminum and transition metals are selected from Group 4
with titanium and zirconium being the most preferred. The organo
portion of the organometallic compound contains functional groups
that are reactive with the functional groups of the polymer
surface. For purposes of the present invention, "organometallic"
compounds are defined as including compounds that do not
necessarily contain metal-carbon bonds. Examples of suitable organo
groups of the organometallic compound are dialkylamide and alkoxide
groups containing from 1 to 18, preferably 2 to 8 carbon atoms.
Examples of alkoxide groups include ethoxide, propoxide,
isopropoxide, butoxide, isobutoxide and tert-butoxide. Examples of
dialkylamide groups include diethyl amide, dipropyl amide,
diisopropyl amide, dibutyl amide, diisobutyl amide and ditert-butyl
amide.
[0031] Reaction may be by transition metal coordination followed by
proton transfer to a dialkylamide or alkoxide ligand and loss of an
dialkylamine or alkanol, so that an organometallic layer covalently
bonds with the polymer surface. The organometallic layer is
believed to consist of a transition metal layer in which the
dialkylamide or alkoxide groups of the transition metal are
covalently bonded to the polymer at the reactive functional
group,
[0032] Depending upon the position of the transition metal on the
Periodic Chart, the transition metal dialkylamide or alkoxide will
have from two to six dialkylamide or alkoxide groups. Transition
metal tetra-alkoxides and tetra-dialkylamides are preferred, with
the most preferred transition metal tetra-alkoxide and
tetra-dialkylamide being zirconium tetra tert-butoxide and
zirconium tetra-diethylamide.
[0033] With Group 4 transition metal tetra-alkoxides and
tetra-dialkylamides, at least one of the alkoxide or amide groups
reacts with reactive functional groups on the polymer surface to
form covalent bonds between the reactive functional groups and the
transition metal. The reaction proceeds by proton transfer to a
dialkylamide or alkoxide group of a transition metal, producing an
equivalent quantity of the corresponding dialkyl amine or alkanol.
At least one dialkylamide or alkoxide group does not react and
remains available for subsequent reaction with the organic
overlayer material.
[0034] Group 5 transition metals form pentaalkoxides or
pentadialkylamides and oxotrialk-oxides or oxotridialkylamides that
are suitable for use with the present invention. These compounds
can also react by proton transfer where possible to covalently bond
the transition metal to the reactive functional group on the
polymer surface, producing an equivalent quantity of an alkanol or
dialkylamine and leaving at least one unreacted alkoxide or
dialkylamide group for subsequent reaction with organic overlayer
material.
[0035] Group 6 transition metals form hexaalkoxides or
hexadialkylamides, oxotetra-alkoxides or oxotetradialkylamides and
dioxo-dialkoxides or dioxodidialkylamides that are all suitable for
use with the present invention. These compounds can also react by
proton transfer where possible to covalently bond the transition
metal to the reactive functional group on the polymer surface,
producing an equivalent quantity of an alkanol or dialkylamine and
leaving at least one unreacted alkoxide or dialkylamide group for
subsequent reaction with organic overlayer material.
[0036] Advantageously, many of the transition metal alkoxides
dialkylamides suitable for use with the present invention are
commercially available. This includes the preferred zirconium tetra
(tert-butoxide) and zirconium tetra-diethylamide, which may be
obtained from Strem. However, the transition metal alkoxides and
dialkylamides may also be prepared by conventional techniques by
reacting a halide or oxo-halide of the selected transition metal,
depending on the desired number of alkoxide or dialkylamide groups,
with the corresponding alkoxide or dialkylamide of a metal selected
from Group 1 or Group 2 of the Periodic Chart.
[0037] With regard to the preferred metals titanium and zirconium,
the alkoxides are titanates and zirconates. These compounds can be
reactive simple esters, polymeric forms of the esters and chelates
that are relatively stable. Examples of various compounds
include
[0038] a. alkyl ortho esters of titanium and zirconium having the
general formula M(OR).sub.4, wherein M is selected from Ti and Zr
and R is C.sub.1-18 alkyl,
[0039] b. polymeric alkyl titanates and zirconates obtainable by
condensation of the alkyl ortho esters of (a), i.e., partially
hydrolyzed alkyl ortho esters of the general formula
RO[-M(OR).sub.2O--].sub.x-1R, wherein M and R are as above and x is
a positive integer,
[0040] c. titanium chelates, derived from ortho titanic acid and
polyfunctional alcohols containing one or more additional hydroxyl,
keto, carboxyl or amino groups capable of donating electrons to
titanium. These chelates have the general formula
Ti(O).sub.a(OH).sub.b(OR').sub.c(XY).sub.d wherein a=4-b-c-d;
b=4-a-c-d; c=4-a-b-d; d=4-a-b-c; R' is H, R as above or X--Y,
wherein X is an electron donating group such as oxygen or nitrogen
and Y is an aliphatic radical having a two or three carbon atom
chain such as [0041] i. --CH.sub.2CH.sub.2--, e.g., of
ethanolamine, diethanolamine and triethanolamine, ##STR2## [0042]
ii. e.g., of lactic acid, ##STR3## [0043] iii. e.g., of
acetylacetone enol form, and ##STR4## [0044] iv. e.g., as in
1,3-octyleneglycol
[0045] d. titanium acylates having the general formula
Ti(OCOR).sub.4-n(OR).sub.n wherein R is C.sub.1-18 alkyl as above
and n is an integer of from 1 to 3, and polymeric forms thereof,
and [0046] e. mixtures thereof.
[0047] The organometallic compound is usually dissolved or
dispersed in a diluent. Examples of suitable diluents are alcohols
such as methanol, ethanol and propanol, aliphatic hydrocarbons,
such as hexane, isooctane and decane, ethers, for example,
tetrahydrofuran and dialkylethers such as diethylether. Also,
adjuvant materials may be present in the organometallic
composition. Examples include surfactants and anti-static agents.
The adjuvants if present are present in amounts of up to 30 percent
by weight based on the non-volatile content of the composition.
[0048] The concentration of the organometallic compound in the
composition is not particularly critical but is usually at least
1.0 micromolar, typically from about 1.0 micromolar to about 100
millimolar, and more typically from about 1.0 micromolar to about
50 millimolar.
[0049] The organometallic treating composition can be obtained by
mixing all of the components at the same time with low shear mixing
or by combining the ingredients in several steps. The
organometallic compounds are reactive with moisture, and care
should be taken that moisture is not introduced with the diluent or
adjuvant materials and that mixing is conducted in a substantially
anhydrous atmosphere.
[0050] The organometallic composition is applied to the polymer
surface by conventional means such as dipping or spraying. The
organometallic compound is then exposed to conditions sufficient to
form a polymeric metal oxide coating preferably with unreacted
dialkylamide or alkoxide and/or hydroxyl groups. This can be
accomplished by depositing the film under conditions resulting in
hydrolysis and self-condensation of the alkoxide or dialkylamide.
These reactions result in a polymeric coating being formed that
provides cohesive strength to the film. The conditions necessary
for these reactions to occur is to deposit the film in the presence
of water, such as a moisture-containing atmosphere. The resulting
film preferably has some unreacted dialkylamide or alkoxide groups
and/or hydroxyl groups for subsequent reaction and possible
covalent bonding with reactive groups of an overlayer material.
Concurrently with the self-condensation reaction, the diluent is
evaporated. Depending on the reactivity of the functional groups in
the organometallic compound and on the polymer surface, heating may
be required to bond the organometallic layer to the substrate. For
example, temperatures of about 50 to about 200.degree. C. may be
used. However, for readily co-reactive groups, ambient
temperatures, that is, about 20.degree. C., may be sufficient.
[0051] As mentioned above, an overlayer can be applied to the
organometallic film. Such an overlayer material can be derived from
a compound, oligomer or polymer that contains groups that are
reactive with the dialkylamide or alkoxide and/or hydroxyl groups.
Preferred overlayers are the layers of organic ligands of
carboxylic and organophosphorus acids as generally described in
U.S. Pat. No. 6,645,644, the disclosure of which is incorporated by
reference. Certain organic components can also enhance the
non-fouling characteristics of a surface so that cells (e.g., from
bacteria, scar tissue, mildew, mold, and other unwanted organisms)
do not adhere well to the treated surface.
[0052] Examples of organophosphorus acids or derivative thereof are
organophosphoric acids, organophosphonic acids and/or
organophosphinic acids including derivatives thereof. Examples of
derivatives are materials that perform similarly as the acid
precursors such as acid salts, acid esters and acid complexes. The
organo group of the phosphorus acid may be a monomeric, oligomeric
or polymeric group. Examples of monomeric phosphorus acids are
phosphoric acids, phosphonic acids and phosphinic acids including
derivatives thereof.
[0053] Examples of monomeric phosphoric acids are compounds or a
mixture of compounds having the following structure:
(RO).sub.xP(O)(OR').sub.y wherein x is 1-2, y is 1-2 and x+y=3, R
is a radical having a total of 1-30, preferably 6 to 18 carbons,
where R' is H, a metal such as an alkali metal, for example, sodium
or potassium or lower alkyl having 1 to 4 carbons, such as methyl
or ethyl. Preferably, for a portion of the phosphoric acid
compounds in the overlayer, R' is H. The organic component of the
phosphoric acid (R) can be a hydrocarbon and can be aliphatic
(e.g., alkyl having 2-20, preferably 6-18 carbon atoms) including a
saturated or unsaturated carbon chain (e.g., an olefin),
unsubstituted or substituted aliphatic, such as fluoro-substituted,
or can be aryl or aryl-substituted moiety. Substitution in the
omega position is preferred.
[0054] Examples of monomeric phosphonic acids are compounds or
mixture of compounds having the formula: ##STR5## wherein x is 0-1,
y is 1, z is 1-2 and x+y+z is 3. R and R'' are each independently a
radical having a total of 1-30, preferably 6-18 carbons. R' is H, a
metal, such as an alkali metal, for example, sodium or potassium or
lower alkyl having 1-4 carbons such as methyl or ethyl. Preferably,
for at least a portion of the phosphonic acid compounds in the
overlayer, R' is H. The organic component of the phosphonic acid (R
and R'') can be a hydrocarbon and can be aliphatic (e.g., alkyl
having 2-20, preferably 6-18 carbon atoms) including a saturated or
unsaturated carbon chain (e.g., an olefin), unsubstituted or
substituted aliphatic such as fluoro-substituted, or can be an aryl
or aryl-substituted moiety. Substitution in the omega position is
preferred.
[0055] Examples of monomeric phosphinic acids are compounds or
mixture of compounds having the formula: ##STR6## wherein x is 0-2,
y is 0-2, z is 1 and x+y+z is 3. R and R'' are each independently
radicals having a total of 1-30, preferably 6-18 carbons. R' is H,
a metal, such as an alkali metal, for example, sodium or potassium
or lower alkyl having 1-4 carbons, such as methyl or ethyl.
Preferably, for at least a portion of the phosphinic acid compounds
in the overlayer, R' is H. The organic component of the phosphinic
acid (R, R'') can be a hydrocarbon and can be aliphatic (e.g.,
alkyl having 2-20, preferably 6-18 carbon atoms) including a
saturated or unsaturated carbon chain (e.g., an olefin), an
unsubstituted or substituted aliphatic such as fluoro-substituted,
or can be an aryl or aryl-substituted moiety. Substitution in the
omega position is preferred.
[0056] Examples of organo groups which may comprise R and R''
include long and short chain aliphatic hydrocarbons, aromatic
hydrocarbons and substituted aliphatic hydrocarbons and substituted
aromatic hydrocarbons. Examples of substituents include carboxyl
such as carboxylic acid, hydroxyl, amino, imino, amido, thio,
cyano, and halo such as fluoro.
[0057] In addition to the organophosphorus acids mentioned above,
oligomeric or polymeric organophosphorus acids resulting from
self-condensation of the respective organophosphorus acid may be
used.
[0058] The overlayer material can further include a suitable
solvent. For example, for organophosphorus materials, solvents such
as an alcohol (e.g., ethanol), tetrahydrofuran, dichloromethane,
chloroform, 2:1 by volume ethanol:toluene, acetonitrile and water
can be used. The concentration of the overlayer material can range
from about 0.1 micromolar to as high as the upper limit of the
solubility of the overlayer material in a specific solvent, for
example, from about 0.1 micromolar to about 100 millimolar, from
about 0.1 micromolar to about 10.0 millimolar, for example, about
1.0 millimolar.
[0059] The solution of the overlayer material can be applied to the
organometallic coating using one or more techniques, and allowing
the solution to evaporate. For example, the solution can be sprayed
(e.g., a few microgram per square centimeter) onto, dropped on,
and/or painted on the organometallic coating. Alternatively, the
substrate with the organometallic coating can be dipped into the
solution. The solution can be applied by doctor blade, reverse
roll, die coater, wire bar, knife and blade coaters. Direct
gravure, micro gravure and reverse gravure techniques can also be
used. Suitable solvents include solvents in which the organic
compound is soluble including aqueous buffer solutions,
tetrahydrofuran, aceto-nitrile, methylene chloride, chloroform and
water, and the like. The substrate will then be removed from the
solution, rinsed with an inert solvent such as water, acetonitrile,
methanol, tetrahydrofuran or the like and then dried to provide a
substrate with a polymer surface having an organic ligand surface
layer covalently attached thereto.
[0060] Besides the techniques mentioned above, the transition metal
amide or alkoxide may be applied to the polymer surface by vapor
deposition. Typically, an excess of transition metal amide or
alkoxide is employed, and the reaction performed at ambient
temperature. With vapor deposition, upon completion of the
reaction, the vacuum should once again be maintained to remove
excess transition metal alkoxide and alkanol byproduct.
[0061] Other methods of applying the solution to the organometallic
coating are described in US 2004/0023048 and PCT/US/2003/034909,
the disclosures of both of which are incorporated by reference.
Application of the solution may be in one or multiple layers.
[0062] After the solution of the overlayer material is applied to
the organometallic coating and the solvent is allowed to evaporate,
the applied layer may optionally be treated to enhance bonding
directly to the organometallic coating. The applied layer can be
treated with heat and/or electromagnetic radiation, such as
microwave radiation (e.g., 2450 MHz or a wavelength of about 12
cm). In some embodiments, the applied layer is exposed to radiant
and/or induction heating, for example, to a temperature of
50.degree. C. to about 200.degree. C. (e.g., about 150.degree. C.)
for about 30-120 seconds. The heating time may be a function of the
temperature used, and the temperature used may be restricted by
design considerations and/or materials limitations.
[0063] In a similar aspect, the present invention can be looked at
as bonding organic ligands or groups to substrates with polymer
surfaces having functional groups, for example amide groups. For
purposes of the present invention, "polymer surfaces having
functional groups" are defined as polymers with functional groups
either within or between monomeric repeating units, a portion of
which are exposed at the polymer surface, either in the form of a
coating or a molded article, and are reactive with the
organometallic compound, for example, a transition metal. Reaction
may be by transition metal coordination to the functional group.
For an amide, this may be followed by N--H proton transfer to an
alkoxide ligand and loss of an alkanol, so that an organometallic
layer covalently bonds with the amide group on the polymer
surface.
[0064] Although not to be bound by any theory, it is believed the
reaction of the transition metal complex with the polymer substrate
proceeds by coordination of the metal to the functional group of
the polymer. This may be followed by the transfer of a proton from
the polymer functional to the ligand of the transition metal,
forming the corresponding dialkylamine or alkanol and bonding the
polymer functional group to the transition metal. In the case of
vapor deposition techniques, once the reaction is complete, the
vacuum is maintained in order to draw off any excess of the
transition metal or dialkylamine or alkanol byproduct.
[0065] Further, although not to be bound by any theory, it is
believed the reaction of the transition metal complex with the
organic overlayer proceeds by coordination of the functional group
on the organic overlayer to the transition metal. This may be
followed by the transfer of a proton from the organic functional to
the ligand of the transition metal, forming the corresponding
dialkylamine or alkanol and bonding the organic overlayer to the
transition metal.
[0066] The organic compound may also be selected from phosphoric
acid and organo-phosphonic acids. For purposes of the present
invention "phosphoric acid" is defined according to its
well-understood meaning, H.sub.3PO.sub.4. "Organophosphonic acids"
refers to compounds having the formula H.sub.2RPO.sub.3, wherein R
is a hydrocarbon ligand with a carbon directly bonded to
phosphorus.
[0067] Overlayers based on organophosphorus acids are covalently
bonded as phosphate esters of the organometallic layer transition
metal. The phosphate esters may be hydrolyzed to form transition
metal polyphosphate coatings on the substrate surface. The
phosphate and polyphosphate coatings are rich in hydroxyl groups
that are available for further chemical modification.
[0068] The coated polymer substrates also include organic ligands
or groups covalently bonded to the polymer surfaces. By reacting
organic overlayer materials with the transition metal dialkylamide
or alkoxide layer, organic ligands form as a layer on the polymer
surface, covalently bonded at the transition metal to the polymer
surface. The ability to react the organic overlayer material with
the transition metal dialkylamide or alkoxide layer covalently
bonded to the polymer surface at ambient temperature is
particularly useful for the attach-ment of biologically active
ligands such as peptides, proteins, or any other ligand which is
deactivated under extreme conditions.
[0069] Examples of biologically active ligands that are covalently
attached to the polymer surface of a substrate by an organometallic
layer include integrins, integrin receptors, cell attachment
mediators, such as peptides containing variations of the
Arg-Gly-Asp integrin binding sequence known to enhance cellular
attachment, and substances that enhance or exclude particular
varieties of cellular or tissue ingrowth. Such substances include,
for example, osteoinductive substances, such as bone morphogenic
proteins (BMP), and substances that induce cellular growth,
proliferation, and/or differentiation such as epidermal growth
factor (EGF), fibroblast growth factor (FGF), platelet-derived
growth factor (PDGF), insulin-like growth factor (IGF-I and II),
TGF-, vascular endothelial growth factor (VEGF) and the like. Other
biologically active ligands include SMAD3, AXIN2, ID2, HEME
Oxygenase-1 and Nell-1. Antibodies, including monoclonal
antibodies, may also be covalently bound to the polymer
surface.
[0070] When desired, an active agent (or a combination of active
agents) can be bound to the polymer surface of a substrate by the
organometallic layer according to the invention in order to
accomplish any of a variety of goals. The particular active
agent(s) used, as well as the mechanism to chemically and/or
physically attach the active agent(s) to the derivatized surface,
will obviously depend upon the chemical and/or physical nature of
the derivatization of the surface, e.g., its reactivity, its
functionality, its surface roughness, etc. Nevertheless, the
following list of active agents that are suitable for surface
immobilization according to the invention is merely exemplary and
should not be construed as being complete.
[0071] In one embodiment, the active agent can include
antileukotrienes or leukotriene receptor antagonists (e.g., for B4,
C4, D4, and/or E4 leukotriene receptors) including, but not limited
to, zafirlukast, montelukast, pranlukast, iralukast, pobilukast, or
the like, or sombinations thereof, and/or salts thereof (e.g.,
Montelukast sodium, which is commercially available under the
tradename SINGULAIR.RTM.).
[0072] In another embodiment, the active agent can include
antihistamines including, but not limited to, ethanolamines (e.g.,
diphenhydramine and/or salts including hydro-chloride,
dimenhydrinate, carbinoxamine, clemastine and/or salts such as
fumarate, bromodiphenhydramine and/or salts such as hydrochloride,
phenytoloxamine, doxyl-amine, or the like, or other salts thereof,
or combinations thereof), ethylenediamines (e.g., tripelennamine
and/or salts such as hydrochloride, pyrilamine and/or salts such as
maleate, antazoline and/or salts such as phosphate, methapyriline,
or the like, or other salts thereof, or combinations thereof),
alkylamines (e.g., chlorpheniramine and/or salts such as maleate,
brompheniramine and/or salts such as maleate, dexchlorpheniramine
and/or salts such as maleate, dimethindene and/or salts such as
maleate, triprolidine and/or salts such as hydrochloride,
pheniramine and/or salts such as maleate, or the like, or other
salts thereof, or combinations thereof), piper-zines (e.g.,
cyclizine and/or salts such as hydrochloride and/or lactate,
meclizine and/or salts such as hydrochloride, hydroxyzine and/or
salts such as hydrochloride and/or pamoate, buclizine,
chlorcyclizine, or the like, or other salts thereof, or
combinations thereof), phenothiazines (e.g., promethazine and/or
salts such as hydro-chloride, propiomazine, methdilazine,
trimeprazine and/or salts such as tartrate, or the like, or other
salts thereof, or combinations thereof), and/or miscellaneous
others (e.g., cyproheptadine, ketotifen, azatadine and/or salts
such as maleate, terfenadine, fexofenadine, astemizole,
diphenylpyraline, phenindamine, or the like, or salts thereof, or
combinations thereof).
[0073] In another embodiment, the active agent can include
antiseptics including, but not limited to, iodine, chlorhexidine
acetate, sodium hypochlorite, and calcium hydroxide.
[0074] In another embodiment, the active agent can include
steroidal anti-inflammatory agents including, but not limited to,
betamethasone, triamcinolone, dexamethasone, prednisone,
mometasone, fluticasone, beclomethasone, flunisolide, budesonide,
or the like, or salts thereof, or combinations thereof. In another
embodiment, the active agent can include non-steroidal
anti-inflammatory agents including, but not limited to, fenoprofen,
flurbiprofen, ibuprofen, ketoprofen, naproxen, oxaprozin,
diclofenac, etodolac, indomethacin, ketorolac, nabumetone,
sulindac, tolmetin, meclofenamate, mefenamic acid, piroxicam,
suprofen, or the like, or salts thereof, or combinations
thereof.
[0075] In another embodiment, the active agent can include
decongestants including, but not limited to, ephedrine,
phenylpropanolamine, pseudoephedrine, phenylephrine, epinephrine,
ephedrine, desoxyephedrine, naphazoline, oxymetazoline,
tetrahydrozoline, xylometazoline, propylhexedrine, or the like, or
salts thereof, or combinations thereof.
[0076] In another embodiment, the active agent can include
mucolytics including, but not limited to, acetylcysteine, dornase
alpha, or the like, or salts thereof, or combinations thereof.
[0077] In another embodiment, the active agent can include
anticholinergics including, but not limited to, ipratropium,
atropine, scopolamine, or the like, or salts thereof, or
combinations thereof.
[0078] In another embodiment, the active agent can include
non-antibiotic antimicrobials including, but not limited to,
taurolidine or the like.
[0079] In another embodiment, the active agent can include mast
cell stabilizers including, but not limited to, cromolyn,
nedocromil, ketotifen, salts thereof (e.g., sodium), or
combinations thereof.
[0080] In another embodiment, the active agent can include one or
more active ingredients such as anti-infective agents,
anti-inflammatory agents, mucolytic agents, antihistamines,
antileukotrienes, decongestants, anticholinergics, antifungals, and
combinations of these classes of agents. Anti-infective agents
contemplated by the present invention include, but are not limited
to antibiotics, anti-virals, non-antibiotic antimicrobials, and
antiseptics. Anti-inflammatory agents contemplated by the present
invention include, but are not limited to steroidal and
non-steroidal anti-inflammatory agents, and mast cell inhibitors.
Antifungal agents contemplated by the present invention include,
but are not limited to amphotericin B, and azole antifungals.
Examples of contemplated antibiotics include, but are not limited
to cefuroxime, ciprofloxacin, tobramycin, cefoperazone,
erythromycin, and gentamycin.
[0081] Exemplary anti-infective agents include, but are not limited
to, penicillins, cephalosporins, macrolides, ketolides,
sulfonamides, quinolones, aminoglycosides, beta lactam antibiotics,
and linezolid. Exemplary non-antibiotic antimicrobials include
taurolidine. Exemplary steroidal anti-inflammatory agents include
glucocorticoids. Exemplary nonsteroidal anti-inflammatory agents
include diclofenac. Exemplary mast cell stabilizers include
cromolyn and nedcromil sodium. Exemplary mucolytic agents are
acetylcysteine and dornase alpha. Exemplary decongestants are
phenylephrine, naphazoline, oxymetazoline, tetrahydrozoline and
xylometoazoline. Exemplary antihistamines include loratidine.
Exemplary antibiotic combinations include cefuroxime and
gentamicin. Exemplary anticholinergics include ipratropium,
atropine and scopolamine. Exemplary antifungals include
amphotericin B, itraconazole, fluconazole, and miconazole.
[0082] In another embodiment, the active agent can include, but are
not limited to, anti-inflammatory agents (e.g., alclometasone,
amcinonide, amlexanox, balsalazide, betamethasone, celecoxib,
choline magnesium, trisalicylate, choline salicylate, chlobetasol,
colchicine, cortisone acetate, curcumin, disunite, dexamethasone,
diclofenac, diflunisal, etodolac, fenoprofen, fluocinolone,
fluometholone, flurandren-olide, flurandrenolide, flurbiprofen,
hydrocortisone, ibuprofen, indomethacin, ketoprofen, ketorolac,
meclofenamate, mefenamic acid, meloxicam, mesalamine,
Methylprednisolone, nabumetone, naproxen, olsalazine, oxaprozin,
piroxicam, prednisone, rofecoxib, salsalate, sulfasalazine,
sulindac, tolmetin, triamcinolone, valdecoxiband,
analogs/derivatives thereof, salts thereof, or combinations
thereof), immunosuppressants (e.g., azathioprine, basiliximab,
cyclosporine, daclizumab, leflunomide, lymphocyte immune globulin,
methotrexate, muromonab-CD3, myco-phenolate, sirolimus, tacrolimus,
thalidomideand, analogs/derivatives thereof, salts thereof, or
combinations thereof), anti-cell proliferation agents (e.g.,
alkylating agents such as busulfan, cisplatin, cyclophosphamide,
oxaliplatin, or the like; nitrosourea compounds such as in
carmustine, lomustine, or the like; anthracycline compounds such as
epirubicin, mitoxantrone, or the like; anti-androgen compounds such
as bicalutamide, flutamide, nilutamide, or the like; antibiotics
such as bleomycin, dactinomycin, mitomycin, or the like;
anti-metabolite compounds such as cladribine, flurouracil,
gemcitabine, hydroxyurea, methotrexate, or the like;
anti-microtubular compounds such as docetaxel, paclitaxel, or the
like; aromatase inactivators such as anastrozole, exemestane, or
the like; hormones such as estramumustine, megestrol, or the like;
monoclonal antibody compounds such as alemtuzumab, rituximab, or
the like; protein synthesis inhibitors such as asparaginase,
pegaspargase, or the like; other compounds such as carboplatin,
dipyridamole, doxorubin, doxorubicin, etopo-side, imatinib,
misonidazole, mercaptopurine, testolactone, trimetrexate,
glucuronate, tiripazamine, topotecan, vindesine, vincristine,
analogs/derivatives thereof, salts thereof, or combinations
thereof), anti-thromosis, anti-platelet, and/or fibrinolysis agents
(e.g., abcimab, antithrombin III, argatroban, aspirin, clopidogrel,
dipyridamole, eptifibatide, fondaparinux, heparin, low molecular
weight heparin, heparin sulfate, recombinant hirudin such as
bivalirudin, lepirudin, or the like, ticlopidine, tissue
recombinant plasminogen activators such as alteplase, reteplase,
streptokinase, tenecteplase, urokinase, or the like, tirofibanand,
analogs/derivatives thereof, salts thereof, or combinations
thereof), extracellular matrix mediators (e.g., calprotectin,
catechins, sulfonylated amino acid hydroxamates, tetracycline
compounds such as demeclo-cycline, doxycycline, minocycline,
oxytetracycline, tetracycline, or the like, analogs or derivatives
thereof, salts thereof, or combinations thereof), and the like, and
combinations thereof.
[0083] In another embodiment, the active agent can include, but are
not limited to, anti-thrombotic agents such as heparin, heparin
derivatives, urokinase, PPack (dextro-phenylalanine proline
arginine chloromethylketone), or the like, analogs/derivatives
thereof, salts thereof, or combinations thereof; steroidal and
non-steroidal anti-inflammatory agents (NSAIDs) such as
dexamethasone, prednisolone, cortico-sterone, hydrocortisone and
budesonide estrogen, sulfasalazine and mesalamine, salicylic acid,
salicylates, ibuprofen, naproxen, sulindac, diclofenac, piroxicam,
ketoprofen, diflunisal, nabumetone, etodolac, oxaprozin,
indomethacin, or the like, analogs/derivatives thereof, salts
thereof, or combinations thereof; anti-neoplastic or
anti-proliferative or anti-mitotic agents such as paclitaxel,
5-fluorouracil, cisplatin, vinblastine, vincristine, epothilones,
endostatin, angiostatin, doxorubicin, metho-trexate, angiopeptin or
the like, analogs/derivatives thereof, salts thereof, or
combinations thereof, monoclonal antibodies capable of blocking
smooth muscle cell proliferation, thymidine kinase inhibitors, or
the like, analogs/derivatives thereof, salts thereof, or
combinations thereof; anesthetic agents such as lidocaine,
bupivacaine, ropivacaine, or the like, analogs/derivatives thereof,
salts thereof, or combinations thereof; anti-coagulants such as
D-Phe-Pro-Arg chloromethyl ketone, an RGD peptide-containing
compound, heparin, hirudin, antithrombin compounds, platelet
receptor antagonists, antithrombin antibodies, anti-platelet
receptor antibodies, aspirin, prostaglandin inhibitors, platelet
inhibitors, tick antiplatelet peptides, or the like,
analogs/derivatives thereof, salts thereof, or combinations
thereof; vascular cell growth promoters such as growth factors,
transcriptional activators, translational promoters, or the like,
analogs/derivatives thereof, salts thereof, or combinations
thereof; vascular cell growth inhibitors such as growth factor
inhibitors, growth factor receptor antagonists, transcriptional
repressors, translational repressors, replication inhibitors,
inhibitory antibodies, antibodies directed against growth factors,
bifunctional molecules consisting of a growth factor and a
cytotoxin, bifunctional molecules consisting of an antibody and a
cytotoxin, or the like, analogs/derivatives thereof, salts thereof,
or combinations thereof; protein kinase and tyrosine kinase
inhibitors such as tyrphostins, genistein, quinoxalines, or the
like, analogs/derivatives thereof, salts thereof, or combinations
thereof; prostacyclin analogs; cholesterol-lowering agents;
angiopoietins; resveratrol and derivatives thereof; antimicrobial
agents such as triclosan, cephalosporins, .beta.-lactams,
aminoglycosides, nitrofurantoin, or the like, analogs/derivatives
thereof, salts thereof, or combinations thereof; cytotoxic agents;
cytostatic agents; cell proliferation affectors; vasodilating
agents; agents that interfere with endogenous vascoactive
mechanisms; analogs/derivatives thereof; salts thereof; metabolites
thereof; or combinations thereof.
[0084] Exemplary genetic active agents include, but are not limited
to, anti-sense DNA and RNA as well as DNA coding for: (a)
anti-sense RNA, (b) tRNA or rRNA to replace defective or deficient
endogenous molecules, (c) angiogenic factors including growth
factors such as acidic and basic fibroblast growth factors,
vascular endothelial growth factor, epidermal growth factor,
transforming growth factor .alpha. and .beta., platelet-derived
endothelial growth factor, platelet-derived growth factor, tumor
necrosis factor .alpha., hepatocyte growth factor and insulin-like
growth factor, (d) cell cycle inhibitors including CD inhibitors,
and (e) thymidine kinase ("TK") and other agents useful for
interfering with cell proliferation. Also of interest is DNA
encoding for the family of bone morphogenic proteins ("BMP's"),
including BMP-2, BMP-3, BMP-4, BMP-5, BMP-6 (Vgr-1), BMP-7 (OP-1),
BMP-8, BMP-9, BMP-10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, and
BMP-16. Such molecules include any of the "hedgehog" proteins, or
the DNA's encoding them.
[0085] Vectors of interest for delivery of genetic active agents
include, but are not limited to, (a) plasmids, (b) viral vectors
such as adenovirus, adeno-associated virus, lentivirus, or the
like, and (c) non-viral vectors such as lipids, liposomes, cationic
lipids, or the like.
[0086] Cells include cells of human origin (autologous or
allogenic), including stem cells, or from an animal source
(xenogenic), which can be genetically engineered if desired to
deliver proteins of interest.
[0087] Non-limiting examples of useful antimicrobial agents
include: Antiamebics, e.g., Arsthinol, Bialamicol, Carbarsone,
Cephaeline, Chlorbetamide, Chloroquine, Chlorphenoxamide,
Chlortetracycline, Dehydroemetine, Dibromopropamidine, Diloxanide,
Diphetarsone, Emetine, Fumagillin, Glaucarubin, Glycobiarsol,
8-Hydroxy-7-iodo-5-quinoline-sulfonic Acid, Iodochlorhydroxyquin,
lodoquinol, Paromomycin, Phanquinone, Polybenzarsol, Propamidine,
Quinfamide, Scenidazole, Sulfarside, Teclozan, Tetracycline,
Thiocarbamizine, Thiocarbarsone, Tinidazole; Antibiotics, e.g.
Amino-glycosides (such as Amikacin, Apramycin, Arbekacin,
Bambermycins, Butirosin, Dibekacin, Dihydrostreptomycin,
Fortimicin(s), Gentamicin, Isepamicin, Kaniamycin, Micronomicin,
Neomycin, Neomycin Undecylenate, Netilmicin, Paromomycin,
Ribo-stamycin, Sisomicin, Spectinomycin, Streptomycin, Tobramycin,
Trospectomycin, and the like), Amphenicols (such as Azidamfenicol,
Chloramphenicol, Florfenicol, Thiamphenicol, and the like),
Ansamycins (such as Rifamide, Rifampin, Rifamycin, Rifapentine,
Rifaximin, and the like), .beta.-Lactams (e.g., Carbacephems,
Loracarbef, Carbapenems (such as Biapenem, Imipenem, Meropenem,
Panipenem, and the like), Cephalosporins (such as Cefaclor,
Cefadroxil, Cefamandole, Cefatrizine, Cefazedone, Cefazolin,
Cefcapene Povoxil, Cefclidin, Cefdinir, Cefditoren, Cefepime,
Cefetamet, Cefixime, Cefinenoxine, Cefodizime, Cefonicid,
Cefoperazone, Ceforanide, Cefotaxime, Cefotiam, Cefozopran,
Cefpimizole, Cefpiramide, Cefpirome, Cefpodoxime Proxetil,
Cefprozil, Cefroxadine, Cefsulodin, Ceftazidime, Cefteram,
Ceftezole, Ceftibuten, Ceftizoxime, Ceftriaxone, Cefuroxime,
Cefuzonam, Cephacetrile Sodium, Cephalexin, Cephaloglycin,
Cephaloridine, Cephalosporin, Cephalothin, Cephapirin Sodium,
Cephradine, Pivcefalexin, and the like), Cepha-mycins (such as
Cefbuperazone, Cefmetazole, Cefminox, Cefotetan, Cefoxitin, and the
like), Monobactams (such as Aztreonam, Carumonam, Tigemonam, and
the like), Oxacephens (such as Flomoxef, Moxalactam, and the like),
Penicillins (such as Amdinocillin, Amdinocillin Pivoxil,
Amoxicillin, Ampicillin, Apalcillin, Aspoxicillin, Azidocillin,
Azlocillin, Bacampicillin, Benzylpenicillic Acid, Benzylpenicillin
Sodium, Carbenicillin, Carindacillin, Clometocillin, Cloxacillin,
Cyclacillin, Dicloxacillin, Epicillin, Fenbenicillin, Floxacillin,
Hetacillin, Lenampicillin, Metampicillin, Methicillin Sodium,
Mezlocillin, Naacillin Sodium, Oxacillin, Penamecillin, Penethamate
Hydr-iodide, Penicillin G Benethamine, Penicillin G Benzathine,
Penicillin G Benzhydryl-amine, Penicillin G Calcium, Penicillin G
Hydrabamine, Penicillin G Potassium, Penicillin G Procaine,
Penicillin N, Penicillin O, Penicillin V, Penicillin V Benzathine,
Penicillin V Hydrabamine, Penimepicycline, Phenethicillin
Potassium, Pipera-cillin, Pivampicillin, Propicillin, Quinacillin,
Sulbenicillin, Sultamicillin, Talampicillin, Temocillin,
Ticarcillin, and the like), Ritipenem, Lincosamides (such as
Clindamycin, Linco-mycin, and the like), Macrolides (such as
Azithromycin, Capbomycin, Clarithromycin, Dirithromycin,
Erythromycin, Erythromycin Acistrate, Erythromycin Estolate,
Erythromycin Glucoheptonate, Erythromycin Lactobionate,
Erythromycin Propionate, Erythromycin Stearate, Josamycin,
Leucomycins, Midecamycins, Miokamycin, Oleandomycin, Primycin,
Rokitamycin, Rosaramicin, Roxithromycin, Spiramycin,
Troleandomycin, and the like), poly-peptides (such as Amphomycin,
Bacitracin, Capreomycin, Colistin, Enduracidin, Envio-mycin,
Fusafungine, Gramicidin S, Gramicidin(s), Mikamycin, Polymyxin,
Pristinamycin, Ristocetin, Teicoplanin, Thiostrepton,
Tuberactinomycin, Tyrocidine, Tyrothricin, Vanco-mycin, Viomycin,
Virginiamycin, Zinc Bacitracin, and the like), Tetracyclines (such
as Apicycline, Chlortetracycline, Clomocycline, Demeclocycline,
Doxycycline, Guamecycline, Lymecycline, Meclocycline, Methacycline,
Minocycline, Oxytetracycline, Penimepi-cycline, Pipacycline,
Rolitetracycline, Sancycline, Tetracycline, and the like),
Cyclo-serine, Mupirocin, Tuberin; synthetic antibacterial agents,
e.g. 2,4-Diaminopyrimi-dines (such as Brodimoprim, Textroxoprim,
Trimethoprim, and the like), Nitrofurans (such as Furaltadone,
Furazolium Chloride, Nifuradene, Nifuratel, Nifurfoline,
Nifur-pirinol, Nifurprazine, Nifurtoinol, Nitrofirantoin, and the
like), Quinolones and Analogs (such as Cinoxacin, Ciprofloxacin,
Clinafloxacin, Difloxacin, Enoxacin, Fleroxacin, Flumequine,
Grepafloxacin, Lomefloxacin, Miloxacin, Nadifloxacin, Nadilixic
Acid, Norflaxacin, Ofloxacin, Oxolinic Acid, Pazufloxacin,
Pefloxacin, Pipemidic Acid, Piromidic Acid, Rosoxacin, Rufloxacin,
Sparfloxacin, Temafloxacin, Tosufloxacin, Trovafloxacin, and the
like), Sulfonamides (such as Acetyl Sulfamethoxpyrazine,
Benzylsulfamide, Chloramine-B, Chloramine-T, Dichlor-amine T,
N2-Formylsulfisomidine, N4- -D-Glucosylsulfanilamide, Mafenide,
4'-(Methyl-sulfamoyl)sulfanil-anilide, Noprylsulfainide,
Phthalylsulfacetamide, Phthalylsulfathiazole, Salazo-sulfadimidine,
Succinylsulfathiazole, Sulfabenzamide, Sulfacetamide,
Sulfachlor-pyridazine, Sulfachrysoidine, Sulfacytine, Sulfadiazine,
Sulfadicramide, Sulfadi-methoxine, Sulfadoxine, Sulfaethidole,
Sulfaguanidine, Sulfaguanol, Sulfalene, Sulfaloxic, Sulfamerazine,
Sulfameter, Sulfamethazine, Sulfamethizole, Sulfa-methomidine,
Sulfamethoxazole, Sulfamethoxypyridazine, Sulfametrole,
Sulfamido-chrysoidine, Sulfamoxole, Sulfanilamide,
4-Sulfanilamidosalicylic Acid, N4-Sulfanilyl-sulfanilamide,
Sulfanilylurea, N-Sulfanilyl -3,4-xylamide, Sulfanitran,
Sulfaperine, Sulfaphenazole, Sulfaproxyline, Sulfapyrazine,
Sulfapyri-dine, Sulfasomizole, Sulfa-symazine, Sulfathiazole,
Sulfathiourea, Sulfatolamide, Sulfisomi-dine, Sulfisoxazole, and
the like), Sulfones (such as Acedapsone, Acediasulfone,
Acetosul-fone Sodium, Dapsone, Diathymosulfone, Glucosulfone
Sodium, Solasulfone, Succisulfone, Sulf-anilic Acid,
p-Sulfanilylbenzylamine, Sulfoxone Sodium, Thiazolsulfone, and the
like), Clofoctol, Hexedine, Methenamine, Methenamine
Anhydromethylenecitrate, Methenamine Hippurate, Methenamine
Mandelate, Methenamine Sulfosalicylate, Nitroxoline, Taurolidine,
Xibomol, and the like; leprostatic antibacterial agents, such as
Acedapsone, Acetosulfone Sodium, Clofazimine, Dapsone,
Diathymosulfone, Glucosulfone Sodium, Hydnocarpic Acid,
Solasulfone, Succisulfone, Sulfoxone Sodium, and the like,
antifungal agents such as Allyl-amines Butenafine, Naftifine,
Terbinafine, Imidazoles (e.g., Bifonazole, Butoconazole,
Cholordantoin, Chlormid-azole, Cloconazole, Clotrimazole,
Econazole, Enilconazole, Fenticonazole, Flutrimazole, Isoconazole,
Ketoconazole, Lanoconazole, Miconazole, Omoconazole, Oxiconazole
Nitrate, Sertaconazole, Sulconazole, Tioconazole, and the like),
Thiocarbamates (e.g., Tolcilate, Tolindate, Tolnaftate, and the
like), Triazoles (e.g., Fluconazole, Itraconazole, Saperconazole,
Terconazole, and the like), Acrisorcin, Amorolfine, Biphenamine,
Bromosalicylchloranilide, Buclosamide, Calcium Propionate,
Chlorphenesin, Ciclopirox, Cloxyquin, Coparaffinate, Diamthazole
Dihydrochloride, Exalamide, Flucytosine, Halethazole, Hexetidine,
Loflucarban, Nifuratel, Potassium Iodide, Propionic Acid,
Pyrithione, Salicylanilide, Sodium Propionate, Sulbentine,
Tenonitrozole, Triacetin, Ujothion, Undecylenic Acid, Zinc
Propionate, etc.; or the like; analogs/derivatives thereof; salts
thereof; or combinations thereof.
[0088] Other antimicrobial agents useful in the present invention
include, but are not limited to, Q-lactamase inhibitors (e.g.
Clavulanic Acid, Sulbactam, Tazobactam, and the like);
Chloramphenicols (e.g. Azidamphenicol, Chloramphenicol,
Thiaphenicol, and the like); Fusidic Acid; synthetic agents such as
Trimethoprim, (optionally in combination with sulfonamides)
Nitroimidazoles (e.g., Metronidazole, Tinidazole, Nimor-azole, and
the like), and the like; Antimycobacterial agents (e.g.,
Capreomycin, Clofazimine, Dapsone, Etham-butol, Isoniazid,
Pyrazinamide, Rifabutin, Rifampicin, Streptomycin, Thioamides, and
the like); Antiviral agents (e.g., Acryclovir, Amanta-dine,
Azidothymidine, Ganciclovir, Idoxuridine, Tribavirin, Trifluridine,
Vidarabine, and the like); Interferons; antiseptic agents (e.g.,
Chlorhexidine, Gentian violet, Octenidine, Povidone Iodine,
Quaternary ammonium compounds, Silver sulfadi-azine, Triclosan, and
the like); or the like; analogs/derivatives thereof; salts thereof;
or combinations thereof.
[0089] In some embodiments, the active agent may include, but is
not limited to, collagen (e.g., Type 1), osteonectin, bone
sialoproteins (Bsp), alpha-2HS-glycoproteins, bone Gla-protein
(Bgp), matrix Gla-protein, bone phosphoglycoprotein, bone
phosphor-protein, bone proteoglycan, protolipids, bone morphogenic
proteins (e.g., BMP-1, -2A, -2B, -3, -3b, -4, -5, -6, -7, -8, -8b,
-9, -10, -11, -12, -13, -14, -15), cartilage induction factor,
platelet derived growth factor (PDGF-1, -2), endothelial cell
growth factors (ECGF-1, -2a, -2b), skeletal growth factor
(SKF=IGF-2), insulin-like growth factors (IGF-1, IGF-2), fibroblast
growth factor (ODGF-1, -2, -3, -4, -5, -6, -7, -8, -9, -10, -11,
-12, -13, ,-14, -15, -16, -17, -18, -19, -20, -21, -22, -23),
colony stimulating factor, transforming growth factor (e.g.,
TGF-.alpha., TGF-.beta., or the like), vascular endothelial growth
factors (VEGF), growth/differentiation factors (GDF-1, -3, -5, -6,
-7, -8, -9, -9B, -10, -11, -15, -16), osteogenic proteins
(OP-1=BMP-7, OP-2=BMP-8, OP-3=BMP-8b), bone growth hormone,
parathyroid hormone (PTH), insulin, calcitonin, and the like, and
combinations thereof. Additionally or alternately, the active
agents may include proteins associated with cartilage, such as
chondrocalcining protein; proteins associated with dentin, such as
phosphophoryn, glycoproteins and Gla proteins; proteins associated
with enamel such as amelognin and enamelin; structural proteins
such as fibrin, fibrinogen, keratin, tubulin, elastin, and the
like; blood proteins, whether in plasma or serum, e.g., serum
albumin; non-protein growth factors such as prostaglandins and
statins (e.g., Simvastatin, Lovastatin, or the like); or the like;
analogs/derivatives thereof; salts thereof; or combinations
thereof.
[0090] In another embodiment, the active agent can include amino
acids, anabolics, analgesics and antagonists, anesthetics,
angiogenesis agents, anti-angiogenetic agents, antihelmintics,
anti-adrenergic agents, anti-asthmatics, anti-atherosclerotics,
antibacterials, anticholesterolics, anticholinergics,
anti-coagulants, antidepressants, antidotes, anti-emetics,
anti-epileptic drugs, anti-fibrinolytics, antihistamines,
anti-inflammatory agents, antihyper-tensives, antimetabolites,
antimigraine agents, antimycotics, antinauseants, antineoplastics,
anti-obesity agents, anti-Parkinson agents, antiprotozoals,
antipsychotics, antirheumatics, antiseptics, antivertigo agents,
antivirals, appetite stimulants, bacterial vaccines, bioflavonoids,
calcium channel blockers, capillary stabilizing agents, coagulants,
corticosteroids, detoxifying agents for cytostatic treatment,
diagnostic agents (like contrast media and radio-isotopes), drugs
for treatment of chronic alcoholism, drugs targeting dopaminergic
pathways, electrolytes, enzymes, enzyme inhibitors, ferments,
ferment inhibitors, gangliosides and ganglioside derivatives,
hemostatics, hormones, hormone antag-onists, hypnotics,
immuno-modulators, immunostimulants, immuno-suppressants, minerals,
muscle relaxants, neuron-modulators, neurotransmitters and
neurotropics, osmotic diuretics, parasympatholytics,
para-sympathomimetics, peptides, proteins, psychostimulants,
respiratory stimulants, sedatives, serum lipid reducing agents,
smooth muscle relaxants, sympatholytics, sympathomimetics,
vasodilators, vaso-protectives, vectors for gene therapy, viral
vaccines, viruses, vitamins, oligonucleo-tides and derivatives, or
the like, or analogs/derivatives thereof, salts thereof, and/or
combinations thereof.
[0091] In another embodiment, the active agent can include
antimicrobial agents, analgesics, anti-inflammatory agents, counter
irritants coagulation modifying agents, diuretics,
sympatho-mimetics, anorexics, antacids and other gastrointestinal
agents, antiparasitics, antidepressants, antihypertensives,
anticholinergics, stimulants, antihormones, central and respiratory
stimulants, drug antagonists, lipid-regulating agents, uricosurics,
cardiac glycosides, electrolytes, ergot and derivatives thereof,
expectorants, hypnotics and sedatives, antidiabetic agents,
dopaminergic agents, antiemetics, muscle relaxants,
para-sympathomimetics, anticonvul-sants, antihist-amines,
beta-blockers, purgatives, antiarrhytmics, contrast materials,
radio-pharmaceuticals, antiallergic agents, tranquilizers,
vasodilators, antiviral agents, and anti-neoplastic or cytostatic
agents or other agents with anticancer properties, vitamins
(including micro- and macro-nutrients), or a combination
thereof.
[0092] In another embodiment, the active agent includes an
anti-muscle spasm agent, anti-spasmodic, bone resorption inhibitor,
smooth muscle contractile agent, calcium absorption enhancer,
muscle relaxant, or a mixture thereof. Suitable anti-muscle spasm
agents include, but are not limited to, baclofen, botulinum toxin,
carisoprodol, chlorphenesin, chlorzoxazone, cyclobenzaprine,
dantrolene, diazepam, metaxalone, methocarbamol, orphenadrine,
tizani-dine, and mixtures thereof. Suitable anti-spasmodics
include, but are not limited to, atropine, baclofen, dicyclomine,
hyoscine, propatheline, oxybutynin, S-oxybutynin, tizanidine,
cevim-eline, chlordiazepoxide, hydrochloride, dicyclomine,
hyoscine, hyoscyamine, glycopyrrolate, and mixtures thereof.
Suitable bone resorption inhibitors include, but are not limited to
alendronate, ibandronate, minodronate, risedronate, etidronate,
tiludronate, and mixtures thereof. A suitable smooth muscle
contractile agent includes, but is not limited to, hyoscine, and
mixtures thereof. Suitable calcium absorption enhancers include,
but are not limited to, alfacalcidol, calcitriol, and mixtures
thereof. Suitable muscle relax-ants include, but are not limited
to, baclofen, carisoprodol, chlorphenesin, chlorzoxa-zone,
cyclobenzaprine, dantro-lene, diazepam, metaxalone, methocarbamol,
orphenadrine, and mixtures thereof.
[0093] In another embodiment, the active agent includes an
anti-diuretic, anti-muscle spasm agent, anti-spasmodic, agent for
treating urinary incontinence, anti-diarrheal agent, agent for
treating nausea and/or vomiting, smooth muscle contractile agent,
anti-secretory agent, enzyme, anti-ulcerant, bile acid replacement
and/or gallstone solubilizing drug, or a mixture thereof. Suitable
anti-diuretics include, but are not limited to, acetazolamide,
benzthiazide, bendroflumethazide, bumetanide, chlorthali-done,
chlorothiazide, ethacrynic acid, furose-mide, hydrochlorothiazide,
hydroflume-thiazide, methyclothiazide, polythiazide, quinetha-zone,
spironolactone, triamterene, torsemide, trichlomethiazide,
desmopressin, oxytocin, and mixtures thereof. Suitable anti-muscle
spasm agents include, but are not limited to, baclofen, botulinum
toxin, carisoprodol, chlorphenesin, chlorzoxazone, cyclobenzaprine,
dantrolene, diazepam, metaxalone, methocarbamol, orphenadrine,
tizanidine, and mixtures thereof. Suitable anti-spasmodics include,
but are not limited to, atropine, baclofen, dicyclo-mine, hyoscine,
propatheline, oxybutynin, S-oxybutynin, tizanidine, and mixtures
thereof. Suitable agents for treating urinary incontinence include,
but are not limited to, darifenacin, vamic-amide, detrol, ditropan,
imipramine, and mixtures thereof. Suitable anti-diarrheal agents
include, but are not limited to, ondansetron, palno-setron,
tropisetron, attapulgite, atropine, bismuth, diphenoxylate,
loperamide, and mixtures thereof. Suitable agents for treating
nausea and/or vomiting include, but are not limited to, alosetron,
dolasetron, granisetron, meclizine, metoclopramide, ondansetron,
palnosetron, prochloperazine, promethazine, trimethobenzamiode,
tropisetron, and mixtures thereof. A suitable smooth muscle
contractile agent includes, but is not limited to, hyoscine.
Suitable anti-secretory agents include, but are not limited to,
esomeprazole, lansoprazole, omeprazole, pantoprazole, rabeprazole,
tenetopra-zole, ecabet, misoprostol, teprenone, and mixtures
thereof. Suitable enzymes include, but are not limited to,
alpha-galactosidase, alpha-L-iduronidase, imiglucerase/alglucerase,
amylase, lipase, protease, pancreatin, olsalazine, and mixtures
thereof. Suitable anti-ulcerants include, but are not limited to,
cimetidine, ranitidine, famotidine, misoprostol, sucralfate,
pantopra-zole, lansoprazole, omepra-zole, and mixtures thereof. A
suitable bile acid replacement and/or gallstone solubilizing drug
includes, but is not limited to, ursodiol.
[0094] In another embodiment, the active agent includes an
endocrine modulator, glucose production inhibitor, agent for
treatment of type II diabetes, anti-secretory agent, glycolipid,
glycoprotein, anti-hyperthyroid agent, thyroid hormone, or a
mixture thereof. Suitable endocrine modulators include, but are not
limited to, methimazole, voglibose, finasteride, GI198745,
liothyronine, glyburide, metformin, nateglinide, ioglitazone,
pegvisomant, minoxidil, and mixtures thereof. Suitable glucose
production inhibitors include, but are not limited to, acarbose,
acetohexamide, chlorpropamide, glipizide, glyburide, metformin,
miglitol, nateglinide, pioglitazone, rosiglitazone, tolbutamide,
tolazamide, and mixtures thereof. Suitable agents for treatment of
type II diabetes include, but are not limited to, acarbose,
acetohex-amide, chlorpropamide, glipizide, glyburide, metformin,
miglitol, nateglinide, pioglit-azone, rosiglitazone, tolbutamide,
tolazamide, and mixtures thereof. Suitable anti-secretory agents
include, but are not limited to, esomeprazole, lansoprazole,
omep-razole, pantoprazole, rabeprazole, tenetoprazole, ecabet,
misoprostol, teprenone, and mixtures thereof. Suitable glycolipids
include, but are not limited to imigulcerase, vanco-mycin, vevesca
(OGT 918), GMK vaccine, and mixtures thereof. Suitable
glycoproteins include, but are not limited to, staphvax,
bimosiamose (TBC 1269), GCS-100, heparin, and mixtures thereof.
Suitable anti-hyperthyroid agents include, but are not limited to,
methimazol, propylthiouracil, and mixtures thereof.
[0095] In another embodiment, the active agent includes a
cholesterol-lowering agent, aldo-sterone antagonist,
triglyceride-lowering agent, leukotriene receptor antagonist,
immuno-modulator or immunogen, glucose production inhibitor, agent
for treatment of type II diabetes, bone resorption inhibitor,
calcium absorption enhancer, insulin enhancing agent, insulin
sensitizer, cytokine, metabolic regulator, mast cell mediator,
eosinophil and/or mast cell antagonist, glycolipid, glycoprotein,
anti-inflammatory drug, anti-obesity drug, COX (cyclooxygenase)
and/or LO (lipoxygenase) inhibitor, or a mixture thereof. Suitable
cholesterol-lowering agents include, but are not limited to,
atorvastatin, benzofibrate, bezafibrate, cerivastatin,
cholestyramine, ciprofibrate, clofibrate, colesevelam, colestipol,
ezetimibe, fluvastatin, gemfibrozil, lovastatin, niacin/lovastatin,
pravastatin, probucol, rosuvastatin, and simvastatin. A suitable
aldosterone antagonist includes, but is not limited to,
spironolactone. A suitable triglyceride-lowering agent includes,
but is not limited to, fenofibrate. Suitable immunomodulators or
immunogens include, but are not limited to, interferon beta 1A,
interferon beta 1B. Suitable glucose production inhibitors include,
but are not limited to, acarbose, acetohexamide, chlorpropamide,
glipizide, glyburide, metformin, miglitol, nateglinide,
pioglitazone, rosiglitazone, tolbutamide, and tolazamide. Suitable
insulin enhancing agents include, but are not limited to,
acamprosate, miglitol, troglitazone, chlorpropamide, glimepiride,
glipizide, glyburide, and repagli-nide. A suitable insulin
sensitizer includes, but is not limited to, is BRL 49653. Suitable
cytokines include, but are not limited to, darbepoetin alfa,
epoetin alpha, erythropoietin, and NESP. Suitable metabolic
regulators include, but are not limited to, allopurinol and
oxypurinol. A suitable eosinophil and/or mast cell antagonists
includes, but is not limited to, nedocromil. Suitable
anti-inflammatory drugs include, but are not limited to, alosetron,
anakinra, beclomethasone, betamethasone, budesonide, clobetasol,
celecoxib, cromolyn, desoximetasone, dexametha-sone, epinastic,
etanercept, etoricoxib, flunisolide, fluocinonide, fluticasone,
formoterol, hydrocortisone, hydroxychloroquine, ibudilast,
ketotifen, meloxicam, mesalamine, metho-trexate,
methylprednisolone, mometasone, montelukast, nedocromil,
olsalazine, prednisone, ramatroban, rofecoxib, salsalate,
terbutaline, triamcinolone, valdecoxib, and zafirlukast. Suitable
anti-obesity drugs include, but are not limited to, dexedrine,
diethylpropion, mazindol, oleoyl-estrone, phentermine,
phendimetrazine, and sibutramine. A suitable COX and/or LO
inhibitor includes, but is not limited to, is ML-3000.
[0096] In another embodiment, the active agent includes an
anti-arrhythmic, anti-hyper-tensive, heart regulator,
cardiovascular agent, plaque stabilization agent, vasodilator,
anti-anginal, anti-coagulant, anti-hypotensive, anti-thrombotic,
drug for treating congestive heart failure, p-FOX (fatty acid
oxidation) inhibitor, or a mixture thereof. Suitable
anti-arrhythmics include, but are not limited to, adenosine,
amioda-rone, bepridil, bretylium, digitoxin, digoxin, diltiazem,
disopyramide, dofetilide, D-sotolol, flecainide, lidocaine,
mexiletine, milrinone, phenyloin, pilsicainide, procain-amide,
propafenone, propranolol, quinidine, tocainide, dofetilide, and
mixtures thereof. Suitable anti-hypertensives include, but are not
limited to, acebutolol, alfuzosin, amlodipine, atenolol,
amlodipine/benazepril, barnidipine benazepril, bepridil, betaxolol,
bisoprolol, bosentan, candesartan, captopril, cariporide,
carvedilol, celiprolol, cilazapril, clonidine, diltiazem,
doxazosin, enalapril, eplerenone, eprosartan, esmolol, felodipine,
fenoldopam, fosinopril, guanfacine, imidapril, irbesartan,
isradipine, labetalol, lercanidipine, lisinopril, losartan,
manidipine, methyldopa, metoprolol, moxonidine, nadolol,
nicardipine, nicorandal, nifedipine, nitrendipine, nosoldipine,
omapatrilat, perindopril erbumine, pindolol, prazosin, propranolol,
quinapril, ramipri, sotalol, spirapril, tamsulosin, telmisartan,
terazosin, torsemide, trandolapril, valsartan, vatanidipine,
midodrine, and mixtures thereof. Suitable heart regulators include,
but are not limited to, digoxin, digitoxin, dobut-amine, and
mixtures thereof. Suitable cardiovascular agents include, but are
not limited to, edaravone, iloprost, levosimendan, molsidomine,
tezosentan, tirilazad, YM087, adenosine, avasimibe, fenofibrate,
and mixtures thereof. A suitable plaque stabilization agent
includes, but is not limited to, avasimibe. Suitable vasodilators
include, but are not limited to, buflomedil, cilostazol,
dipyridamole, diazoxide, hydralazine, minoxidil, naftidrofuryl,
nicorandil, nitroprusside, alprostadil, apomorphine, phentolamine
mesylate, sildenafil, tadalafil, vardenifil, and mixtures thereof.
Suitable anti-anginals include, but are not limited to,
amilodipine, amyl nitrite, atenolol, bepridil, diltiazem,
erythrityl tetranitrate, felodipine, isosorbide dinitrate,
isradipine, metoprolol, nadolol, nicardipine, nifedipine,
nimodipine, pentaerythritol tetranitrate, propranolol, and mixtures
thereof. Suitable anti-coagulants include, but are not limited to,
abciximab, ardeparin, argatroban, bivalirudin, clopidogrel,
dalteparin, danaparoid, desirudin, dipyridamole, enoxaparin,
eptifibatide, fondaparinux, H376/95, lepirudin, melagatran,
nadroparine, nafamostat mesilate, pentosan, pentoxifylline,
reviparin, sarpogrelate, SNAC/SNAD-heparin, ticlopidine,
tinzaparin, tirofiban, warfarin, and mixtures thereof. Suitable
anti-hypotensives include, but are not limited to, midodrine,
dobutamine, fludrocortisone, and mixtures thereof. Suitable
anti-thrombotics include, but are not limited to, aspirin,
abciximab, enoxaparin, integrelin, ticlopidine, and mixtures
thereof. Suitable drugs for treating congestive heart failure
include, but are not limited to, amrinone, benazepril, bumetanide,
captopril, digitoxin, digoxin, dobutamine, dopamine, enalapril,
ethacrynic acid, fosino-pril, furosemide, hydralazine, lisinopril,
milrinone, minoxidil, moexipril, quinapril, ramipril, torsemide,
and mixtures thereof. A suitable p-FOX inhibitor includes, but is
not limited to, ranolazine.
[0097] In another embodiment, the active agent includes an
aldosterone antagonist, immunomodulator or immunogen,
immunosuppressant, cytokine, leukotriene receptor antagonist, mast
cell mediator, eosinophil and/or mast cell antagonist, mucolytic,
glucocorti-coid, glycolipid, or a mixture thereof. A suitable
aldosterone antagonist includes, but is not limited to,
spironolactone. Suitable immuno-suppressants include, but are not
limited to, azathioprine, cyclophosphamide, cyclosporine, ERL 080,
enlimomab, methotrexate, mitoxan-trone, mycophenolate, mofetil,
sirolimus, tacrolimus (FK-506), and mixtures thereof. Suitable
mucolytics for use in the buccal sprays of the invention include,
but are not limited to, ambroxol, bromhexin, fudostein,
acetylcestine, and mixtures thereof.
[0098] In another embodiment, the active compound is a p-FOX (fatty
acid oxidation) inhibitor, acetylcholinesterase inhibitor, nerve
impulse inhibitor, anti-cholinergic, anti-convulsant,
anti-psychotic, anxiolytic agent, dopamine metabolism inhibitor,
agent to treat post stroke sequelae, neuroprotectant, agent to
treat Alzheimer's disease, neurotransmitter, neurotransmitter
agonist, sedative, agent for treating attention deficit disorder,
agent for treating narcolepsy, central adregenic antagonist,
anti-depression agent, agent for treating Parkinson's disease,
benzodiazepine antagonist, stimulant, neurotransmitter antagonist,
tranquilizer, or a mixture thereof. Suitable acetylcholinesterase
inhibitors include, but are not limited to, galantamine,
neostig-mine, physostigmine, and edrophonium. Suitable nerve
impulse inhibitors include, but are not limited to,
levobupivacaine, lidocaine, prilocaine, mepivacaine, propofol,
rapacuronium bromide, ropivacaine, tubocurarine, atracurium,
doxaurium, miva-curium, pancuronium, vercuronium, pipecuronium, and
rocuronium. Suitable anti-cholinergics for use in the buccal sprays
of the invention include, but are not limited to, amantadine,
ipratropium, oxitropium, and dicycloverine. Suitable
anti-convulsants include, but are not limited to, acetazolamide,
carbamazepine, clonazepam, diazepam, dival-proex (valproic acid),
ethosuximide, lamotrignine acid, levetriacetam, oxcarbazepine,
phenol-barbital, phenytoin, pregabalin, primidone, remacemide,
trimethadione, topiramate, vigabatrin, and zonisamide. Suitable
anti-psychotics include, but are not limited to, amisulpride,
aripiprazole bifemelane, bromperidol, clozapine, chlorpromazine,
haloperidol, iloperidone loperidone, olanzapine, quetiapine,
fluphenazine, fumarate, risperidone, thiothixene, thioridazine,
sulpride, and ziprasidone. Suitable anxiolytic agents include, but
are not limited to, amitrypti-line, atracurium, buspirone,
chlorzoxazone, clorazepate, cisatracurium, cyclobenza-prine,
eperisone, esopiclone, hydroxyzine, mirtazapine, mivacurium,
pagoclone, sulperide, zaleplon, and zopiclone. Suitable dopamine
metabolism inhibitors include, but are not limited to, entacapone,
lazebemide, selegiline, and tolcapone. Suitable agents to treat
post stroke sequelae include, but are not limited to, glatiramer,
interferon beta 1A, interferon beta 1B, estradiol, and
progesterone. Suitable neuron-protectants include, but are not
limited to, donepezil, memanine, nimodipine, riluzole,
rivastigmine, tacrine, TAK147, and xaliproden. Suitable agents to
treat Alzheimer's disease include, but are not limited to,
carbidopa, levodopa, tacrine, donezepil, rivastigmine, and
galantamine. Suitable neurotransmitters include, but are not
limited to, acetylcholine, serotonin, 5-hydroxytryptamine (5-HT),
GABA, glutamate, aspartate, glycine, histamine, epinephrine,
norpinephrine, dopamine, adenosine, ATP, and nitric oxide. Suitable
neuron-transmitter agonists include, but are not limited to,
almotriptan, aniracetam, atomoxetine, benserazide, bromocriptine,
bupropion, cabergoline, citalopram, clomipramine, desipramine,
diazepam, dihydroergotamine, doxepin duloxetine, eletriptan,
escitalopram, fluvoxamine, gabapentin, imipramine, moclobemide,
naratriptan, nefazodone, nefiracetam acamprosate, nicergoline,
nortryptiline, paroxetine, pergolide, pramipexole, rizatriptan,
ropinirole, sertraline, sibutramine, sumatriptan, tiagabine,
trazodone, venlafaxine, and zolmitriptan. Suitable sedatives
include, but are not limited to, dexmedetomidine, eszopiclone,
indiplon, zolpidem, and zaleplon. Suitable agents for treating
attention deficit disorder include, but are not limited to,
amphetamine, dextroamphetamine, methyl-phenidate, and pemoline.
Suitable agents for treating narcolepsy include, but are not
limited to, modafinil and mazindol. A suitable central adregenic
antagonist includes, but is not limited to, mesoridazine. Suitable
anti-depression agents include, but are not limited to,
amitriptyline, amoxapine, bupropion, clomipramine, clomipramine,
clorgyline, desipramine, doxepin, fluoxetine, imipramine,
isocarbox-azid, maprotiline, mirtazapine, nefazodone,
nortriptyline, paroxetine, phenelzine, protriptyline, sertraline,
tranylcypromine, trazodone, and venlafaxine. Suitable agents for
treating Parkinson's disease include, but are not limited to,
amantadine, bromocriptine, carvidopa, levodopa, pergolide, and
selegiline. A suitable benzo-diazepine antagonist includes, but is
not limited to, flumazenil. A suitable neuron-transmitter
antagonist includes, but is not limited, to deramciclane. Suitable
stimulants include, but are not limited to, amphetamine,
dextroamphetamine, dinoprostone, methylphenidate, methylphenidate,
modafinil, and pemoline. A suitable tranquilizer includes, but is
not limited to, mesoridazine.
[0099] In another embodiment, the active agent includes a nerve
impulse inhibitor. Suitable nerve impulse inhibitors include, but
are not limited to levobupivacaine, lidocaine, prilocalne,
mep-ivacaine, propofol, rapacuronium bromide, ropivacaine,
tubocurarine, atracurium, doxacurium, mivacurium, pancuronium,
vecuronium, pipecuronium, rocuronium, and mixtures thereof.
[0100] In another embodiment, the active agent includes an
anti-opioid agent. Suitable anti-opioid agents for use in the
buccal sprays of the invention include, but are not limited to,
naloxone, nalmefene, naltrexone, cholecystokinin, nociceptin,
neuropeptide FF, oxytocin, vasopressin, and mixtures thereof.
[0101] In another embodiment, the active agent includes an
anti-migraine agent. Suitable anti-migraine agents for use in the
buccal sprays of the invention include, but are not limited to,
frovatriptan, zolmitriptan, rizatriptan, almotriptan, eletriptan,
naratriptan, almotriptan, ergotamine, diethylergotamine,
sumatriptan, and mixtures thereof.
[0102] In another embodiment, the active agent includes a pain
control agent. Suitable pain control agents for use in the buccal
sprays of the invention include, but are not limited to,
non-steroidal anti-inflammatory drugs, alfentanil, butorphanol,
codeine, dezocine, fentanyl, hydrocodone, hydromorphone,
levorphanol, meperidine, methadone, morphine, nalbuphine,
oxycodone, oxymorphone, propoxyphene, pentazocine, sufentanil,
tramadol, and mixtures thereof.
[0103] In another embodiment, the active agent includes an
anesthetic. Suitable anesthetics for use in the buccal sprays of
the invention include, but are not limited to, benzo-natate,
bupivacaine, desflurane, enflurane, isoflurane, levobupivacaine,
lidocaine, mepivacaine, prilocalne, propofol, rapacuronium bromide,
ropivacaine, sevoflurane, ketamine, and mixtures thereof.
[0104] In another embodiment, the active agent can include, but is
not limited to, cyclosporine, sermorelin, octreotide acetate,
calcitonin-salmon, insulin lispro, sumatriptan succinate,
clozepine, cyclobenzaprine, dexfenfluramine hydrochloride,
glyburide, zidovudine, erythromycin, ciprofloxacin, ondansetron
hydrochloride, dimenhydrinate, cimetidine hydro-chloride,
famotidine, phenyloin sodium, phenyloin, carboprost thromethamine,
carboprost, diphenhydramine hydrochloride, isoproterenol
hydrochlor-ide, terbutaline sulfate, terbutaline, theophylline,
albuterol sulfate, neutraceuticals (i.e., nutrients with
pharmacological action, e.g., carnitine, valerian, echinacea, and
the like), or the like; analogs/derivatives thereof;
salts/alternate salts thereof; or combinations thereof.
[0105] Any opioid or non-.mu.-opioid, a pharmaceutically acceptable
salt thereof, a base form thereof, or mixture of any combination of
such opioids and/or their derivatives that are known in the art can
be included. Opioids believed to have at least some .mu.-opioid
receptor agonist activity (and optionally at least some agonist
activity also at one or more of the .kappa.-opioid receptor, the
.delta.-opioid receptor, and the ORL-1 receptor) include, but are
not limited to, alfentanil, allylprodine, alphaprodine,
anileridine, benzylmorphine, bezitramide, buprenor-phine,
butorphanol, clonitazene, codeine, desomorphine, dextromoramide,
dezocine, diampr-omide, diamorphone, dihydro-codeine,
dihydromorphine, dihydromorphone, dihydroisomor-phine, dimenoxadol,
dimepheptanol, dimethylthiambutene, dioxaphetyl butyrate,
dipipanone, eptazocine, ethoheptazine, ethylmethylthiambutene,
ethylmorphine, etonitazene, etorphine, dihydroetorphine, fentanyl,
heroin, hydrocodone, hydromorphone, hydromorphodone,
hy-droxypethidine, isomethadone, ketobemidone, levorphanol,
levophenacylmorphan, lofentanil, meperidine, meptazinol,
metazocine, methadone, metopon, morphine, myrophine, narceine,
nicomorphine, norlevorphanol, normethadone, nalorphine, nalbuphene,
normorphine, norpip-anone, opium, oxycodone, oxymorphone, pantopon,
papaveretum, paregoric, pentazocine, phenadoxone, phendimetrazine,
phendimetrazone, phenomorphan, phenazocine, pheno-peridine,
piminodine, piritramide, propheptazine, promedol, properidine,
propoxyphene, propylhexedrine, sufentanil, tilidine, tramadol, and
mixtures thereof. Non-.mu.-opioids include, but are not limited to,
ORL-1-specific opioid agonists, such as nociceptin, deltorphin, and
the like, and mixtures thereof. In a preferred embodiment, the
opioid includes buprenorphine, pharmaceutically acceptable salts
thereof, base forms thereof, fentanyl, pharmaceutically acceptable
salts thereof, base forms thereof, oxycodone, pharmaceutically
acceptable salts thereof, base forms thereof, and any combination
of such opioids and/or their derivatives.
[0106] In certain embodiments, the opioid agonist includes
hydrocodone, morphine, hydromorphone, oxycodone, codeine,
levorphanol, meperidine, methadone, oxymorphone, bupren-orphine,
fentanyl, dipipanone, heroin, tramadol, etorphine,
dihydroetorphine, butorphanol, levorphanol, pharmaceutically
acceptable salts thereof, base forms thereof, and any and all
mixtures thereof. The opioid agonist can, in some embodiments,
include oxycodone, hydro-codone, fentanyl, buprenorphine,
pharmaceutically acceptable salts thereof, base forms thereof, and
any and all mixtures thereof. The opioid agonist can, in other
embodiments, include buprenorphine, pharmaceutically acceptable
salts thereof, base forms thereof, fentanyl, pharmaceutically
acceptable salts thereof, base forms thereof, and any combination
of such opioids and/or their derivatives.
[0107] General categories of active agents can, in one embodiment,
include, but are not limited to: ACE inhibitors; adenohypophyseal
hormones; adrenergic neuron blocking agents; adrenocortical
steroids; inhibitors of the biosynthesis of adrenocortical
steroids; alpha-adrenergic agonists; alpha-adrenergic antagonists;
selective alpha-two-adrenergic agonists; androgens; anti-addictive
agents; antiandrogens; anti-infectives, such as antibiotics,
antimicrobials, and antiviral agents; analgesics and analgesic
combinations; anorexics; antihelmintics; antiarthritics;
antiasthmatic agents; anticonvulsants; antidepressants;
antidiabetic agents; antidiarrheals; antiemetic and prokinetic
agents; antiepileptic agents; antiestrogens; antifungal agents;
antihistamines; antiinflammatory agents; antimigraine preparations;
anti-muscarinic agents; antinauseants; antineoplastics;
antiparasitic agents; anti-parkinsonism drugs; antiplatelet agents;
antiprogestins; antipruritics; antipsychotics; anti-pyretics;
antispasmodics; anticholinergics; antithyroid agents; antitussives;
azaspirodecane-diones; sympathomimetics; xanthine derivatives;
cardiovascular preparations, including potassium and calcium
channel blockers, alpha blockers, beta blockers, and
antiarrhythmics; antihypertensives; diuretics and antidiuretics;
vasodilators, including general coronary, peripheral, and cerebral;
central nervous system stimulants; vasoconstrictors; cough and cold
preparations, including decongestants; hormones, such as estradiol
and other steroids, including cortico-steroids; hypnotics;
immunosuppressives; muscle relaxants; parasympatholytics;
psychostimulants; sedatives; tranquilizers; nicotine and acid
addition salts thereof; benzodiaz-epines; barbituates;
benzothiadiazides; beta-adrenergic agonists; beta-adrenergic
antagonists; selective beta-one-adrenergic antagonists; selective
beta-two-adrenergic antagonists; bile salts; agents affecting
volume and composition of body fluids; butyrophenones; agents
affecting calcification; catecholamines; cholinergic agonists;
cholinesterase reactivators; dermatological agents;
diphenylbutyl-piperidines; ergot alkaloids; ganglionic blocking
agents; hydantoins; agents for control of gastric acidity and
treatment of peptic ulcers; hematopoietic agents; histamines;
5-hydroxytryptamine antagonists; drugs for the treatment of
hyper-lipiproteinemia; laxatives; methylxanthines; moncamine
oxidase inhibitors; neuron-muscular blocking agents; organic
nitrates; pancreatic enzymes; phenothiazines; prostaglan-dins;
retinoids; agents for spasticity and acute muscle spasms;
succinimides; thioxanthines; thrombolytic agents; thyroid agents;
inhibitors of tubular transport of organic compounds; drugs
affecting uterine motility; vitamins; and the like; or a
combination thereof.
[0108] Alternately or in addition to an opioid agonist, another
active compound may be added including, but not limited to,
flurogestone acetate, hydroxyprogesterone, hydroxyprogesterone
acetate, hydroxyprogesterone caproate, medroxy-progester-one
acetate, norethindrone, norethindrone acetate, norethisterone,
norethynodrel, desogestrel, 3-keto desogestrel, gestadene,
levonorgestrel, estradiol, estradiol benzoate, estradiol valerate,
estradiol cyprionate, estradiol decanoate, estradiol acetate,
ethynyl estradiol, estriol, estrone, mestranol, betamethasone,
betametha-sone acetate, cortisone, hydrocortisone, hydrocortisone
acetate, corticosterone, fluocinolone acetonide, prednisolone,
prednisone, triamcinolone, aldosterone, androsterone, testosterone,
methyl testosterone, or a combination thereof.
[0109] Alternately or in addition to an opioid agonist, another
active compound may be added including, but not limited to: a)
corticosteroids, e.g., cortisone, hydrocortisone, prednisolone,
beclomethasone propionate, dexamethasone, betamethasone,
flumethasone, triamcinolone, triamcinolone acetonide, fluocinolone,
fluocinolone acetonide, fluocinolone acetate, clobetasol
propionate, or the like, or a combination thereof; b) analgesic
anti-inflammatory agents, e.g., acetaminophen, mefenamic acid,
flufenamic acid, indomethacin, diclofenac, diclofenac sodium,
alclofenac, ibufenac, oxyphenbutazone, phenylbutazone, ibuprofen,
flurbiprofen, ketoprofen, salicylic acid, methylsalicylate,
acetylsalicylic acid, 1-menthol, camphor, slindac, tolmetin sodium,
naproxen, fenbufen, or the like, or a combination thereof; c)
hypnotic sedatives, e.g., phenobarbital, amobarbital,
cyclobarbital, lorazepam, haloperidol, or the like, or a
combination thereof; d) tranquilizers, e.g., fulphenazine,
thiorid-azine, diazepam, flurazepam, chlorpromazine, or the like,
or a combination thereof; e) anti-hypertensives, e.g., clonidine,
clonidine hydrochloride, bopinidol, timolol, pindolol,
propran-olol, propranolol hydrochloride, bupranolol, indenolol,
bucumolol, nifedipine, bunitrolol, or the like, or a combination
thereof; f) hypotensive diuretics, e.g., bendroflumethiazide,
poly-thiazide, methylchlorthiazide, trichlor-methiazide,
cyclopenthiazide, benzyl hydrochloro-thiazide, hydrochlorothiazide,
bumetanide, or the like, or a combination thereof; g) anti-biotics,
e.g., penicillin, tetracycline, oxytetracycline, metacycline,
doxycycline, minocycline, fradiomycin sulfate, erythromycin,
chloramphenicol, or the like, or a combination thereof; h)
anesthetics, e.g., lidocaine, benzocaine, ethylaminobenzoate, or
the like, or a combination thereof; i) antimicrobial agents, e.g.,
benzalkonium chloride, nitrofurazone, nystatin, sulfacet-amide,
clotriamazole, or the like, or a combination thereof; j)
anti-fungal agents, e.g., penta-mycin, amphotericin B, pyrrol
nitrin, clotrimazole, or the like, or a combination thereof; k)
vitamins, e.g., vitamin A, ergocalciferol, cholecalciferol,
octotriamine, riboflavin butyric acid ester, or the like, or a
combination thereof; l) antiepileptics, e.g., nitrazepam,
meprobamate, clonazepam, or the like, or a combination thereof; m)
antihistamines, e.g., diphenhydramine hydrochloride,
chlorphen-iramine, diphenylimidazole, or the like, or a combination
thereof; n) antitussives, e.g., dextromethorphan, terbutaline,
ephedrine, ephedrine hydrochloride, or the like, or a combination
thereof; o) sex hormones, e.g., progesterone, estradiol, estriol,
estrone, or the like, or a combination thereof; p) antidepressants,
e.g., doxepin; q) vaso-dilators, e.g., nitroglycerin, isosorbide
nitrate, nitroglycol, pentaerythritol tetranitrate, dipyridamole,
or the like, or a combination thereof; r) other drugs, e.g.,
5-fluorouracil, dihydroergotamine, desmopressin, digoxin,
methoclopramide, domperidone, scopolamine, scopolamine
hydrochloride, or the like, or a combination thereof; or the like;
or a combination thereof.
[0110] In another embodiment, the active agent can include, but is
not limited to, anti-staphylococcal agents (e.g., YSPXTNF, YSPWTNF,
YSPWTNF-NH2, GENBANK/AF202641, GENBANK/AF205220, GENBANK/AAG03056,
or the like, or combinations thereof). Other agents that modulate
the production or secretion of bacterial or microbial toxins or
virulence factors may also be used as active agents. For instance,
thiolactones and bacterial toxin regulatory proteins such as
RNAIII-inhibiting peptides (RIPs) are classes of active agents.
See, e.g., Balaban, N., et al., "Regulation of Staphylococcus
aureus pathogenesis via target of RNAIII-activating Protein
(TRAP)," J. Biol Chem., 2001 Jan 26; 276(4): 2658-67, which is
incorporated by reference herein in its entirety.
[0111] When an active agent of the present invention is acidic,
salts may be prepared from pharmaceutically acceptable non-toxic
bases. Salts derived from all stable forms of inorganic bases
include aluminum, ammonium, calcium, copper, iron, lithium,
magnesium, manganese, potassium, sodium, zinc, etc. In one
embodiment, the salt includes ammonium, calcium, magnesium,
potassium, or a sodium salt. Salts derived from pharmaceutically
acceptable organic non-toxic bases include salts of primary,
secondary, and tertiary amines, substituted amines including
naturally occurring substituted amines, cyclic amines and basic
ion-exchange resins such as arginine, betaine, caffeine, choline,
N,N dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol,
2-dimethylaminoethanol, ethanolamine, ethylene-diamine,
N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine,
histidine, iso-propylamine, lysine, methyl-glucosamine, morpholine,
piperazine, piperidine, polyamine resins, procaine, purine,
theobromine, triethylamine, trimethylamine, tripropylamine,
etc.
[0112] When an active agent of the present invention is basic,
salts may be prepared from pharmaceutically acceptable non-toxic
acids. Such acids include acetic, benzene-sulfonic, benzoic,
camphorsulfonic, citric, ethane-sulfonic, fumaric, gluconic,
glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic,
mandelic, methane-sulfonic, mucic, nitric, pamoic, pantothenic,
phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, etc.
In one embodiment, the acid includes citric, hydrobromic, maleic,
phosphoric, sulfuric, and/or tartaric acids.
[0113] Although some active agents can be bound directly to the
polymer surface of a substrate by the organometallic layer
according to the invention, many active agents according to the
present invention can often be attached to the polymer surface of a
substrate via .alpha.,.omega.-di-functional linkers or spacers
bound to the organic overlayer. Such spaces or linkers will often
be tailored to the particular active agent(s) used. It is noted
that the - and .omega.-functional groups of the linkers/spacers can
be similar or different, and often are different particularly where
release of the active agent(s) is(are) intended (e.g., in vivo).
Such attachment (and release) of active agents can be effected by
covalent bonding (cleaving), ionic bonding (dissociation), physical
transitions of state such as crystallization (melting) or liquid
crystalline-type ordering (disordering), hydrogen bonding
(dissociation), van der Waals interactions (repul-sions), or the
like, or any combination thereof. The linkers/spacers can be of
relatively small molecular weight (e.g., less than about 200
g/mol), relatively intermediate molecular weight (e.g., from about
200 to about 2000 g/mol), relatively large molecular weight (e.g.,
more than about 2000 g/mol), or some combination thereof.
Particularly when the linkers/spacers are of relatively
intermediate and/or relatively large molecular weight, they can
include, but are not limited to, oligomers, polymers, and/or
copolymers described above as bioactive moieties or biodegrade-able
polymers; additionally or alternately, the linkers/spacers can
include, but are not limited to, oligomers, polymers, and/or
copolymers having one or more of the following types of repeat
units: urethanes, ureas, ethers, ketones, esters, amines,
carbonates, amides, saccharides, or the like, or combinations
thereof.
[0114] In other embodiments, polyfunctional linkers/spacers can be
used to attach active agents to the coatings/derivatized surfaces
according to the invention. Such poly-functional linkers/spacers
can include, but are not limited to, oligomers, polymers, and/or
copolymers that are branched, hyperbranched, dendritic,
star-shaped, brushes, combs, block, multiblock, or the like, or any
combination thereof.
[0115] The transition metal phosphate esters can also serve as a
template for first chemical, then biological growth of bone tissue
hydroxyapatite in the implant surface. Surface-bound transition
metal phosphate layers insinuate themselves directly into bone
tissue hydroxylapatite to make a strong composite seal between the
implant surface and the hydroxyapatite. Alternatively, the
phosphate may be hydrolyzed to form transition metal polyphosphates
having a two-dimensional structure, the layers of which also
insinuate themselves directly into bone tissue hydroxyapatite.
[0116] Essentially any organophosphonic acid capable of forming a
thin film on an organometallic surface is suitable for use with the
present invention. The organophosphonic acid will have a
hydrocarbon ligand that may be saturated or unsaturated, branched
or unbranched, substituted or unsubstituted, and may be aromatic or
non-aromatic. Typical hydrocarbon ligands of organophosphonic acids
will contain between two and twenty carbon atoms or for example,
between three and eighteen carbon atoms. Stearyl ligands, for
example, may be used.
[0117] A preferred class of organophosphorus and organic carboxylic
acids are those with omega-functionalized organo groups that can be
chemically transformed to react and covalently bond to the
aforementioned biologically active and pharmaceutically active
compounds. Examples of such omega functional groups include amino,
carboxylate, thiol, hydroxyl, carbonate, ester, carbamate, and
amide groups.
[0118] A preferred application technique involves a two-stage vapor
deposition process in which the transition metal alkoxide or
dialkylamide is first vapor deposited on the polymer surface. When
the reaction is complete, vacuum is applied to remove excess
transition metal alkoxide and dialkylamine or alkanol by-product,
which is then followed by vapor deposition of the organic overlayer
material. Upon completion of the reaction with the organic
compound, the vacuum is then applied to withdraw excess organic
compound solution and alkanol or dialkylamine by-product.
[0119] Suitable substrates with polymer surfaces also include
fabrics formed from a woven or non-woven fiber. The fiber can be a
natural fiber with exposed functional groups, such as silk, wool,
cotton, collagen, linen, and the like. The fiber can also be a
synthetic fiber with exposed amide groups, such as nylon.
[0120] The polymer-coated and polymer-cast substrates of the
present invention may be fabricated into scaffolds for tissue
engineering and tissue guided regeneration applications, including
reconstructive surgery. The scaffolds may also be molded to form
external scaffolding for the support of in vitro culturing of cells
for the creation of external support organs. The scaffolds may also
be used in transplantation as a matrix for dissociated cells.
[0121] Polymer scaffolds and mixed polymer/ceramic scaffolds for
tissue engineering and their manufacture are known to those skilled
in the art. The scaffold structure is typically porous to allow
generous cellular ingrowth. The polymer scaffolds may also be
molded to form external scaffolding for the support of in vitro
culturing of cells for the creation of external support organs.
[0122] Polymers that are suitable for use as scaffold materials are
non-toxic, physiologically compatible, and optionally
biodegradable. The polymer must be selected for biocompatibility at
the time of implant, and, if biodegradable, the products of its
degradation process must also be biocompatible. Additional
parameters that play an important role include the mechanical
properties of the material, especially its mechanical rigidity.
Relatively high rigidity is advan-tageous so that the scaffold can
withstand the contractile forces exerted by cells growing within
the scaffold. Also important are the thermal properties, such as
the glass transition temperature, T.sub.g, and the biodegradation
kinetics, if degradable, which should match the rate of the healing
process.
[0123] The scaffold functions to mimic the extracellular matrices
(ECM) of the body. The scaffold serves as both a physical support
and an adhesive substrate for isolated cells during in vitro
culture and subsequent implantation. As the transplanted cell
populations grow and the cells function normally, they begin to
secrete their own ECM support. The scaffold polymer preferably
degrades as the need for an artificial support diminishes.
[0124] In the reconstruction of structural tissues like cartilage
and bone, tissue shape is integral to function, requiring the
molding of the polymer scaffold into articles of varying thickness
and shape. Any crevices, apertures or refinements desired in the
three-dimensional structure can be created by removing portions of
the matrix with scissors, a scalpel, a laser beam or any other
cutting instrument. Scaffold applications include the regeneration
of tissues such as nervous, musculoskeletal, cartilaginous,
tendenous, hepatic, pancreatic, ocular, integumentary,
arteriovenous, urinary or any other tissue forming solid or hollow
organs.
[0125] The scaffold may also be used in transplantation as a matrix
for dissociated cells such as chondrocytes or hepatocytes to create
a three-dimensional tissue or organ. Any type of cell can be added
to the scaffold for culturing and implantation, including cells of
the muscular and skeletal systems, such as mesenchymal stem cells,
chondrocytes, fibroblasts, osteocytes and osteoblasts, parenchymal
cells such as hepatocytes, pancreatic cells (including Islet
cells), cells of intestinal origin, and other cells such as
exocrine cells, bile duct cells, parathyroid cells, thyroid cells,
cells of the adrenal-hypothalmic-pituitary axis, heart muscle
cells, kidney epithelial cells, kidney tubular cells, kidney
basement membrane cells, nerve cells, neurons, blood vessel cells,
endothelial cells, cells forming bone and cartilage, smooth muscle
cells, skeletal muscle cells, ocular cells, integumentary cells,
keratinocytes and skin cells, and either as obtained from donors,
embryonic and non-embryonic stem cells, established cell culture
lines, including embryonic and non-embryonic stem cell culture
lines, and either before or after genetic engineering. Pieces of
tissue can also be used, which may provide a number of different
cell types in the same structure.
[0126] The cells are obtained from a suitable donor, or the patient
into which they are to be implanted, dissociated using standard
techniques and seeded onto and into the scaffold. In vitro
culturing optionally may be performed prior to implantation.
Alternatively, the scaffold is implanted, allowed to vascularize,
then cells are injected into the scaffold. Methods and reagents for
culturing cells in vitro and implantation of a tissue scaffold are
known to those skilled in the art. The polymer scaffolds can be
utilized in vivo as tissue engineering and tissue guided
regeneration scaffold in mammals such as primates, including
humans, sheep, horses, cattle, pigs, dogs, cats, rats and mice, or
in vitro.
[0127] The polymer-coated and polymer-cast substrates of the
present invention may also be fabricated from biocompatible
polymers into articles for medical implant applications. The
organic ligand bonded to the article surface can be a
biologically-or pharmaceutically-active compound having utility as
a coating on a medical implant. The polymer-coated and polymer-cast
articles are formed into shaped articles such as vascular grafts
and stents, bone plates, sutures, implantable sensors, barriers for
surgical adhesion prevention, implantable drug delivery devices and
other therapeutic agent articles. The same issues related to
toxicity and tissue compatibility for tissue scaffold polymers also
apply to medical implant polymers.
[0128] The following non-limiting examples set forth hereinbelow
illustrate certain aspects of the present invention. They are not
to be considered limiting as to the scope and nature of the present
invention. In the examples which follow, all parts are by
weight.
EXAMPLES
Example 1
Preparation of RGD-Modified Nylon Substrate
[0129] Activated nylon surface 4 was prepared by first casting
films of 3 (nylon 6/6; (R.dbd.(CH.sub.2).sub.4CO;
R'.dbd.(CH.sub.2).sub.6NH) from 0.1 mM formic acid solution on
glass microscope slides that were rinsed copiously in
Millipore.RTM. water, and evacuated at 10.sup.-2 torr for 3 hours.
The coated slides were then placed in a deposition chamber that was
equipped with two stopcocks for exposure either to vacuum or to
vapor of zirconium tetra(tert-butoxide). The chamber was evacuated
to 10.sup.-3 torr for 30 minutes, and slides of 3 were exposed to
vapor of of zirconium tetra(tert-butoxide) (with external
evacuation) for 30 seconds followed by 5 min exposure without
external evacuation. This cycle was repeated twice, then followed
by an additional 10 minutes of exposure without external
evacuation. The chamber was then evacuated for 16 hours at
10.sup.-3 torr to ensure removal of excess zirconium
tetra(tert-butoxide). The IR spectrum of polymer surface-bound Zr
complex (4) showed .sub..nu.C-H=2976 cm.sup.-1, indicative of
tert-butoxide groups.
[0130] RGD-modified nylon 7a was prepared by immersing a 4-coated
slide in a 0.1 mM solution of phosphonoundecanol (5) in dry THF for
15 min to yield complex 6. Treatment of 6 in a 0.1 mM solution of
3-maleimidopropionic acid N-hydroxysuccinimide ester for 24 hrs
under dry N.sub.2 was followed by copious rinsing successively in
acetonitrile and Millipore.RTM. water and drying in vacuo. As
depicted in FIG. 2, either an RGDC or DANSYL-cys-tethered surface
(7a or 7b) can be obtained via Michael addition, which results in a
complex with a 1:1 ratio of Zr to RGDC or DANSYL-Cys. IR analysis
of 6 showed peaks in the aliphatic region (.sub..nu.CH2, asym=2922
cm.sup.-1; .sub..nu.CH2,sym=2851 cm.sup.-1) characteristic of
disordered alkyl chains. Immersion of 6 in a 0.1 mM aqueous
solution of RGDC at pH 6.5 for 24 hours produced 7a.
Example 2
Preparation of RGD-Modified Nylon Substrate
[0131] RGD-derivatized surface 9a (FIG. 3) was prepared by
immersing a 4-coated slide in a 0.1 mM solution of
3-maleimidopropionic acid N-hydroxysuccinimide ester in dry
acetonitrile for 16 hours to produce 8. Immersion of 8 in a 0.1 mM
aqueous solution of RGDC at pH 6.5 for 24 hours produced 9a. The
nylon-Zr surface complex was derivatized with the succinimide ester
of 3-maleimidopropionic acid directly by transesterification to
produce 8, which can result in either an RGDC or
DANSYL-Cys-tethered surface (9a or 9b). Complexes 9a and 9b have a
1:2 ratio of zirconium to RGDC or DANSYL-Cys, respectively.
Example 3
Preparation of DANSYL-Cys-Modified Nylon Substrate
[0132] Fluorescent molecule-labeled analogues 7b and 9b were
prepared as described for 7a and 9a, but a 0.1 mM aqueous solution
of N-(5-(dimethylamino)-1-naphthyl-sulfonyl)-cysteine (DANSYL-Cys)
was used instead of RGDC (FIGS. 2 and 3).
[0133] To address the issue of solvent-induced polymer swelling,
control films of 3 were prepared by soaking in 0.1 mM DANSYL-cys
solution for 24 hrs. A calibration curve of fluorescence intensity
versus concentration was measured for DANSYL-Cys solutions from
0.16 to 21 .mu.M at pH 7.5 and pH 12.
[0134] Surface complex DANSYL content of 7b and 9b was quantified
by immersion in water at pH 12 for 3 hours, which cleaves the Zr
complexes from the surface, precipitates ZrO.sub.2, and releases
fluorophore from 7b and 9b into solution. The amount of DANSYL
surface-bound through Zr complexes 7b and 9b was measured to be
0.10 nmol/cm.sup.2 and 0.18 nmol/cm.sup.2, respectively. These
amounts are consistent with the DANSYL:Zr stoichiometries of 1:1
and 2:1 indicated for 7b and 9b, respectively (FIGS. 2 and 3).
Notably, they are at least an order of magnitude higher than the
previously highest reported value of about 10 pmol/cm.sup.2 for
polymer surface-derivatization with a peptide. A substantial change
in surface hydrophilicity was confirmed by a decrease in water
contact angle (75.degree. for 3 compared to 50.degree. for 9a).
[0135] The approximate spatial surface coverage by RGD was
calculated from its measured surface loading of 0.2 nmol/cm.sup.2;
assuming an RGD "footprint" of 40 .ANG..sup.2 (determined using
Chem 3D). This corresponds to coverage of about 0.4 cm.sup.2 per
cm.sup.2 of surface, or 40%. With-out being bound by theory, it is
hypothesized that the percent surface coverage by the organic
ligand depends upon the relative sizes of the organic ligand and
the transition metal complex.
Example 4
Hydrolytic Stability of Modified Nylon Substrates
[0136] Nylon films (2 cm.sup.2) derivatized as 7b and 9b and
control films of 3 were immersed in water at pH 7.5 for 7 days at
room temperature, and the supernatants were analyzed by
fluorescence spectroscopy. The samples were then removed from
solution, dried, and immersed in water at pH 12 for 3 hrs, after
which the supernatants were again analyzed by fluorescence
spectro-scopy. Release of DANSYL groups was measured by
fluorescence intensities of supernatants from treated 7b and 9b
which were compared to the control sample (3) over this seven-day
period. Unreacted DANSYLating reagent desorbed from the nylon in
about 3 hours. No release of surface-bound DANSYL material occurred
over the next seven days. Thus zirconium-amide surface-bound
complexes are stable to hydrolysis under these conditions.
Example 5
Cell Response to Surface Modification
[0137] Cell responses to surfaces 3 and 9a were evaluated in vitro.
NIH 3T3 cells were maintained in Dulbecco's Modified Eagle's Medium
(DMEM) with 10% calf serum. Cells were washed with
Phosphate-Buffered Saline (PBS) and released from tissue culture
dishes using 50 mg/mL trypsin and 2 mg/mL EDTA in PBS.
Trypsinization was stopped by washing cells in soybean trypsin
inhibitor (Sigma). Cells were resuspended in serum-free DMEM at
5.times.10.sup.4 cells/mL. Two milliliters of the cell suspension
were added to wells containing the nylon coated surfaces, which had
been pre-blocked for 1 hr in 1% Bovine Serum Albumin. After 90
minutes, non-adherent cells were removed and replaced with fresh,
serum-free DMEM. Cells were fixed, permeabilized, and stained for
the focal adhesion protein vinculin at the indicated time
points.
[0138] Fibroblast cell spreading on (a) nylon 3 control at 3 hrs;
(b) RGD-modified surface 9a at 3 hrs; and (c) RGD-modified surface
9a at 6 hrs is depicted in FIGS. 4a-4c. Staining shows vinculin,
and scale bars are 50 microns. Compared to control untreated nylon
(3), significant numbers of cells attached to the RGDC-modified
surface (9a). Cells were well spread on the RGD-modified surface
showing many focal adhesions when stained with anti-vinculin
antibodies, while the few cells on untreated nylon remained
round.
[0139] Surface-bound Zr-amide complexes, which are readily
synthesized on the surface of nylon 6/6, are thus effective for
activation of that surface for further organic chemical
transformation.
Example 6
Modification of Other Polymer Classes--Polyamide Control
[0140] The ability to modify polyurethanes, polyimides and
polysulfonamides with a transition metal alkoxide with zirconium
tetra(tert-butoxide) was evaluated on a small molecule model from
which the ability to surface-modify the corresponding polymer can
be readily predicted. As a control, the small molecule model was
first tested for the polyamides of Examples 1-5 using
N-hexylacetamide as the model.
[0141] Zr complex 2 (FIG. 1) was prepared by treating
N-hexylacetamide (0.15 g, 1.0 mmol) with Zr tetra(tert-butoxide) 1
(0.40 g, 1.0 mmol) in dry CH.sub.2Cl.sub.2 for 1 hour under
nitrogen. Solvent and reaction byproducts were removed in vacuo to
yield Zr complex 2 in >95% yield as shown by .sup.1H NMR
([CDCl.sub.3]: 0.8 (t, 3H); 1.3 (m, 35H); 1.9 (s, 3H); 3.2
(quartet, 2H)). The reaction proceeded via Zr coordination to the
amide carbonyl, followed by N--H proton transfer to a tert-butoxide
ligand, and loss of tert-butanol. The bonding for the amide moiety
to Zr in 2 (.eta..sup.2-coordination) is indicated by the 8 ppm
downfield shift of the acyl carbon vs. the free amide (.sup.13C NMR
[CDCl.sub.3]: .delta. 170.1 for N-hexyl-acetamide; .delta. 178.1
for 2).
Examples 7-9
Modification of Other Polymer Classes
[0142] The N-hexylacetamide example was repeated in tetrahydrofuran
(THF) at 20 C using methyl N-hexylcarbamate as a model for
polyurethanes, succinimide as a model for poly-imides, and N-hexyl
p-toluene sulfonamide as a model for polysulfonamides. The reaction
progress was observed via .sup.1H NMR and reaction completion was
calculated from the reduction in relative integration of the N--H
NMR peak in each molecule. All reactions were assumed to proceed
via zirconium complex coordination to "carbonyl" oxygen, N--H
proton transfer to ligand, and loss of ligand.
[0143] The succinimide and N-hexyl p-toluene sulfonamide yields
were about 90% after 30 minutes. The methyl N-hexyl carbamate yield
was about 40% after three hours using a stoichiometric excess of
zirconium alkoxide. This is evidence of the ability of transition
metal alkoxides or amides to surface-modify the polymers.
Example 10
Polyurea Modification
[0144] Examples 7-9 were repeated using dicyclohexylurea as a model
for polyurea and tetrakis(diethylamino)zirconium(IV) as the
transition metal alkoxide. The yield was about 90% after 30
minutes. This is indicative of the ability of transition metal
alkoxides or dialkylamides to surface-modify this polymer.
Example 11
Surface Reaction of Collagen with Zirconium
Tetra(tert-butoxide)
[0145] Films of type 1 collagen (from bovine achilles tendon,
Aldrich) were cast from a 0.1 M solution in formic acid on glass
microscope slides that were sonicated in ethanol, rinsed copiously
in Millipore water, and evacuated at 10.sup.-1 torr for 3 h. The
coated slides were then soaked for 24 hours in Millipore water, and
baked at 110.degree. C. for 16 hours to dehydrate them. These
collagen slides were placed in a deposition chamber equipped with
two stopcocks for exposure either to vacuum or to vapor of of
zirconium tetra(tert-butoxide). The chamber was evacuated to
10.sup.-3 torr for 30 min, and slides of collagen were exposed to
vapor of Zr tetra(tert-butoxide) (with external evacuation) for 30
sec followed by 5 min exposure without external evacuation. This
cycle was repeated twice and was then followed by an additional 30
min exposure without external evacuation. The chamber was then
evacuated at 10.sup.-3 torr for 1 h to ensure removal of excess of
zirconium tetra(tert-butoxide), and to give Zr-activated
collagen.
Example 12
1,12-Diphosphonododecane-Modified Collagen
[0146] Collagen was derivatized with 1,12-diphosphonododecane by
immersing a Zr-activated collagen slide in a 0.1 mM solution of
1,12-diphosphonododecane in dry THF for 1 h. The surface was rinsed
copiously in dry THF to produce
1,12-diphosphonododecane-derivatized collagen.
Example 13
11-hydroxyundecylphosphonate-Modified Collagen
[0147] Collagen was derivatized with 11-hydroxyundecylphosphonate
by immersing a Zr-activated collagen slide in a 0.1 mM solution of
11-hydroxyundecylphosphonate in dry THF for 1 h. This surface was
rinsed copiously in dry THF to produce
11-hydroxyundecylphosphonate-derivatized collagen.
Example 14
RGD-Modified Collagen.
[0148] RGD-derivatized collagen was prepared by immersing a
Zr-activated collagen slide in a 0.1 mM solution of
3-maleimidopropionic acid in dry acetonitrile for 1 h to derivatize
the collagen surface with maleimido groups. Immersion of this
maleimido-derivatized collagen in a 0.1 mM aqueous solution of RGDC
at pH 6.5 for 24 h gave RGD-derivatized collagen.
Example 15
Cell Response to Collagen Surface Modification
[0149] Cellular response to collagen coated glass slides modified
with various compounds was assessed by assaying adhesion to
modified and control slides. Collagen coated slides were modified
with 1,12-diphosphonododecane, 11-hydroxyundecylphosphonate and RGD
according to the procedures in Examples 12-14. Bare glass slides
and unmodified collagen coated glass slides were used as negative
controls. Glass slides coated with the cell binding protein
fibronectin were used as positive controls. Gasket sealed tissue
culture chambers were affixed to each slide type and adhesion of
human osteoblasts was assayed after 2 and 24 hour incubation
periods using a Vybrant Cell Adhesion Assay Kit
(invitrogen-V-13181). Representative data is shown in the table
below. TABLE-US-00001 Slide Type Percent Adhesion Glass slide
(-control) 23.8% Collagen coated glass slide (-control) 41.1%
Collagen + 11-hydroxyundecylphosphonate 60.9% Collagen +
1,12-diphosphonododecane 82.9% Collagen + RGD 90.8% Fibronectin
coated slide (+control) 94.9%
* * * * *