U.S. patent application number 11/524926 was filed with the patent office on 2010-09-09 for natural biodegradable matrices and uses thereof.
This patent application is currently assigned to SurModics, Inc.. Invention is credited to Michael J. Burkstrand, Joseph A. Chinn, Stephen J. Chudzik, Dale G. Swan.
Application Number | 20100226960 11/524926 |
Document ID | / |
Family ID | 36958998 |
Filed Date | 2010-09-09 |
United States Patent
Application |
20100226960 |
Kind Code |
A1 |
Chudzik; Stephen J. ; et
al. |
September 9, 2010 |
Natural biodegradable matrices and uses thereof
Abstract
Matrices formed from a natural biodegradable material and that
exhibit surface degradation in the presence of an enzyme are
described. Matrices including a bioactive agent can be implanted or
formed in a subject for release of the bioactive agent upon
degradation of the matrix. The matrix can be provided in various
forms, including coatings for implantable devices, implants, and
in-situ formed matrices. The matrix can also be in the form of a
medical device having a structure that is used to treat a medical
condition.
Inventors: |
Chudzik; Stephen J.; (St.
Paul, MN) ; Chinn; Joseph A.; (Shakopee, MN) ;
Swan; Dale G.; (St. Louis Park, MN) ; Burkstrand;
Michael J.; (Richfield, MN) |
Correspondence
Address: |
Kagan Binder, PLLC
221 Main Street North, Suite 200
Stillwater
MN
55082
US
|
Assignee: |
SurModics, Inc.
|
Family ID: |
36958998 |
Appl. No.: |
11/524926 |
Filed: |
September 21, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60719466 |
Sep 21, 2005 |
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Current U.S.
Class: |
424/426 ;
424/93.1; 514/44R; 514/54 |
Current CPC
Class: |
A61K 9/205 20130101;
A61L 27/58 20130101; A61L 27/54 20130101; A61L 2300/232 20130101;
A61L 29/085 20130101; B05D 1/36 20130101; A61L 2300/258 20130101;
C08B 33/04 20130101; A61L 2300/252 20130101; A61L 31/042 20130101;
A61K 9/0024 20130101; C08B 31/04 20130101; A61L 31/10 20130101;
C08L 3/12 20130101; A61L 29/16 20130101; A61L 2420/02 20130101;
C09D 103/02 20130101; A61K 9/2027 20130101; A61P 43/00 20180101;
C08B 33/02 20130101; A61L 27/20 20130101; A61L 2300/606 20130101;
B05D 7/52 20130101; C09D 103/18 20130101; A61L 27/20 20130101; C08B
31/006 20130101; C08L 3/02 20130101; A61K 9/0051 20130101; A61K
47/34 20130101; A61L 27/34 20130101; A61L 31/16 20130101; A61L
2300/604 20130101; C08L 3/12 20130101 |
Class at
Publication: |
424/426 ; 514/2;
514/44.R; 514/54; 424/93.1 |
International
Class: |
A61F 2/00 20060101
A61F002/00; A61K 38/02 20060101 A61K038/02; A61K 31/7088 20060101
A61K031/7088; A61K 31/715 20060101 A61K031/715; A61K 35/12 20060101
A61K035/12; A61P 43/00 20060101 A61P043/00 |
Claims
1. A method of delivering a bioactive agent to a subject comprising
the steps of: providing a matrix of natural biodegradable material
comprising bioactive agent entrapped within the matrix, wherein the
matrix has a surface, and wherein the matrix is implanted or
immobilized within the subject; and allowing the surface to be
contacted with an enzyme, wherein the matrix prevents the enzyme
from entering the matrix, wherein the enzyme degrades the matrix at
the surface, and wherein bioactive agent is released from the
matrix upon degradation of the natural biodegradable material.
2. The method of claim 1, wherein the matrix is in the form of a
coating on an implantable medical article.
3. The method of claim 1, wherein the matrix is in the form of an
in situ-formed hydrogel.
4. The method of claim 1, wherein the matrix is in the form of an
implantable device.
5. The method of claim 1, wherein the natural biodegradable
material comprises a natural biodegradable polymer.
6. The method of claim 5, wherein the natural biodegradable
material comprises a natural biodegradable polysaccharide.
7. The method of claim 6, wherein the natural biodegradable
polysaccharide is selected from the group consisting of
maltodextrin, polyalditol, and amylose.
8. The method of claim 1, wherein the matrix is formed of
cross-linked natural biodegradable polymers.
9. The method of claim 1, wherein the natural biodegradable
material has a molecular weight of less than 100,000 Da.
10. The method of claim 1, wherein the natural biodegradable
material has a molecular weight of less than 50,000 Da.
11. The method of claim 1, wherein the natural biodegradable
material has a molecular weight in the range of 1000 Da to 10,000
Da.
12. The method of claim 1, wherein the step of allowing, the enzyme
is a carbohydrase.
13. The method of claim 12, wherein the step of allowing, the
enzyme is .alpha.-amylase.
14. The method of claim 1 wherein the bioactive agent is selected
from the group consisting of polypeptides, polynucleotide,
polysaccharides, viral particles, and cells.
15-20. (canceled)
21. The method of claim 1, wherein the natural biodegradable
material has a molecular weight of less than 500,000 Da.
22. The method of claim 1, wherein the natural biodegradable
material has a molecular weight of less than 250,000 Da.
23. The method of claim 1, wherein the matrix is formed from a
composition comprising natural biodegradable polymers comprising
pendent polymerizable groups.
24. The method of claim 23, wherein the pendent polymerizable
groups are selected from the group consisting of vinyl groups,
acrylate groups, methacrylate groups, ethacrylate groups, 2-phenyl
acrylate groups, acrylamide groups, methacrylamide groups,
itaconate groups, and styrene groups.
25. The method of claim 1, wherein the matrix is formed from a
composition comprising natural biodegradable polymers at a
concentration in the range of 5-50% (w/v).
26. The method of claim 25, wherein the matrix is formed from a
composition comprising natural biodegradable polymers at a
concentration in the range of 20-50% (w/v).
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present non-provisional application claims the benefit
of commonly owned provisional Application having Ser. No.
60/719,466, filed on Sep. 21, 2005, and entitled ARTICLES AND
COATINGS INCLUDING NATURAL BIODEGRADABLE POLYSACCHARIDES AND USES
THEREOF.
TECHNICAL FIELD
[0002] The present invention relates to natural biodegradable
matrices and medical uses thereof. Bioactive agents can be included
in the matrices to provide a therapeutic effect to a patient.
BACKGROUND
[0003] Recently, the use of drug-eluting stents (DES) in
percutaneous coronary interventions has received much attention.
DES are medical devices that present or release bioactive agent
into their surroundings (for example, luminal walls of coronary
arteries). Generally speaking, a bioactive agent can be coupled to
the surface of a medical device by surface modification, embedded,
and released from within polymeric materials (matrix-type), or
surrounded by and released through a carrier (reservoir-type). The
polymeric materials in such applications should optimally act as a
biologically inert barrier and not induce further inflammation
within the body. However, the molecular weight, porosity of the
polymer, a greater percentage of coating exposed on the medical
device, and the thickness of the polymer coating can contribute to
adverse reactions to the medical device.
[0004] Another way to deliver bioactive agents from the surface of
a medical device is by using a coating that has a biodegradable
polymer, such as polylactic acid. As the coating degrades, the
bioactive agent is released from the surface of the device.
Although biodegradable coatings that include polylactic acid have
been described in a number of documents, for example, U.S. Pat. No.
6,258,121, there remains a need for improved coatings and coating
materials.
[0005] Some concerns exist that regard the use of biodegradable
materials that degrade into materials that are not typically found
in the body, or that are found at particularly low levels in the
body. These types of biodegradable materials have the potential to
degrade into products that cause unwanted side effects in the body
by virtue of their presence or concentration in vivo. These
unwanted side effects can include immune reactions, toxic buildup
of the degradation products in the liver, or the initiation or
provocation of other adverse effects on cells or tissue in the
body.
[0006] Another problem is that preparations of some biodegradable
materials may not be obtained at consistent purity due to
variations inherent in natural materials. This is relevant at least
with regard to biodegradable materials derived from animal sources.
Inconsistencies in preparations of biodegradable materials can
result in problematic coatings.
[0007] It is also desirable to provide biodegradable drug delivery
coatings that are easy to prepare, cost effective, and that also
offer a wide range of flexibility with regard to the type and
amount of drug or drugs to be delivered from the biodegradable
coating.
[0008] Other aspects of the present invention relate to the use of
polymeric coatings for providing a sealant function to medical
articles. Biodegradable sealant compositions have been used on
articles having porous surfaces, such as fabrics associated with
implantable medical articles. The sealant coating initially renders
the porous surface impermeable to fluids for a period of time.
However, as the sealant materials degrade and are resorbed by the
body, cells involved in tissue repair infiltrate the porous
material and replace the sealant materials. Thus, newly formed
tissue replaces the original function of the coated sealant over a
period of time.
[0009] Animal-derived sealant materials such as collagen and
gelatin are commonly used to coat textile grafts. These materials
can be resorbed in vivo. The blood clotting protein fibrin has also
been utilized as a sealant material. Despite their uses, there are
drawbacks and concerns with using these types of sealant materials.
One particular problem is that it is difficult to produce
consistent sealant compositions from these animal sources due to
batch-to-batch variations inherent in their production.
[0010] In many cases the collagen used in sealant technologies is
obtained from non-human animal sources, such as bovine sources. In
these cases there is the possibility that bovine collagen
preparations may contain unwanted contaminants that are undesirable
for introduction into a human subject. One example of an unwanted
contaminant is the prionic particles that cause Bovine Spongiform
Encephalopathy (BSE).
[0011] BSE, also termed Mad Cow Disease, is one of a group of
progressive neurological diseases called transmissible spongiform
encephalopathies, or TSEs (named for deteriorated areas of the
brain that look like sponges). Various forms of TSE have been
reported, including scrapie in sheep and chronic wasting disease in
elk and mule deer. It is generally believed that the use of
recycled animal parts led to the cross-species contamination of
scrapie in sheep to mad cow disease, and the ingestion of
contaminated beef and bovine products led to the human variant of
this disease, Creutzfeldt-Jakob Disease (CJD).
[0012] Additional concerns are that preparations from animal
sources may provide other unwanted contaminants, such as antigenic
factors. These antigenic factors may promote a localized immune
response in the vicinity of the implanted article and foul its
function. These factors may also cause infection as well as local
inflammation.
[0013] While synthetic materials can be used in the preparation of
medical compositions, these synthetic materials have the potential
of degrading into non-naturally occurring products. These
non-naturally occurring products have the potential to be at least
partially toxic to the organism or immunogenic and cause
inflammation, as well as infection, at or around the site of
implantation.
SUMMARY OF THE INVENTION
[0014] In one aspect, the present invention provides compositions
and methods for preparing biodegradable coatings that are
particularly useful for coating surfaces of implantable medical
devices, such as stents and catheters, and are capable of releasing
bioactive agents from the device surface. These coating
compositions include a natural biodegradable material as a
component that can be crosslinked to form a matrix from which a
therapeutic material such as a drug, a biomolecule, or cells
(referred to herein as a "bioactive agents") can be released or
retained. In some embodiments of the invention, a bioactive agent
is present in and can be released from the biodegradable matrix; in
other embodiments a bioactive agent is present in a biodegradable
microparticle, the microparticle being immobilized within the
matrix.
[0015] In other aspects of the invention, the natural biodegradable
material is used to prepare an article, such as an article that can
be implanted or formed within the body (for example, by in situ
formation). In some aspects, the article can be amorphous, such as
a polymerized mass of natural biodegradable material that is formed
within or on a portion of the body, by using an in vivo
matrix-forming composition.
[0016] In other aspects, the invention provides an article
fabricated from natural biodegradable material, wherein the article
has a defined structure, and wherein the article can be implanted
in the body (such as a filament). Such articles are referred to
herein as "medical implants". A medical implants having a defined
structure can be formed by any suitable process, including molding,
extruding, shaping, cutting, casting, and the like.
[0017] The article can be used for one or more purposes, such as
for releasing or retaining a bioactive agent at a location in the
body. For example, the article can be a bioactive agent-containing
medical implant or depot. The article can also provide one or more
mechanical or physical properties to a portion the body. For
example, the natural biodegradable material can be included in a
composition used for the formation of a biodegradable medical
device such as a stent.
[0018] In some aspects, the article, such as an in vivo formed
matrix, is used in methods for the treatment of any one or more of
a variety of medical conditions or indications, including
restoring, improving, and/or augmenting tissue growth or function,
in particular those for orthopedic, dental, and bone graft
applications. These functions can be provided by placing a
polymerized matrix of biodegradable material in contact with a host
tissue. The matrix can restore or improve tissue growth or function
by, for example, promoting or permitting formation of new tissue
between and into the matrix. The effect on tissue can be caused by
the biodegradable material itself, or the biodegradable material in
combination with one or more bioactive agent(s) that can be present
in and/or released from the matrix. Exemplary bioactive agents that
can affect tissue function include peptides, such as peptides that
are involved in tissue repair processes and belonging to the EGF,
FGF, PDGF, TGF-.beta., VEGF, PD-ECGF or IGF families, and also
peptides derived from bone morphogenetic protein 2, or BMP-2. The
bioactive agent can also be a cell, such as a platelet.
[0019] In another aspect, the invention provides a method of
delivering a bioactive agent to a subject based on the properties
of the biodegradable matrix. The method comprises a step of
providing a matrix comprising natural biodegradable material and
bioactive agent entrapped within the matrix. The matrix has a
surface and is implanted or immobilized within the subject. Another
step of the method includes allowing the surface to be contacted
with an enzyme. The structure of the matrix prevents the enzyme
from entering the matrix. The enzyme, however, degrades the matrix
at the surface and bioactive agent is released from the matrix upon
degradation of the matrix. The matrix provides an improved method
for delivering one or more bioactive agent(s) to a subject with
zero order release rates. In more specific aspects, in the step of
providing, the matrix comprises cross-linked
enzymatically-biodegradable polymers.
[0020] The matrix has a structure that can exclude enzymes selected
from carbohydrases (such as polysaccharidases), proteases,
nucleases, and lipases.
[0021] Another aspect of the invention is directed to a method for
treating a medical condition in a subject. In this method, the
structural property of the medical article is used to treat the
condition. Bioactive agents are not required in the article, but
can be included if desired. The method includes a step of providing
to a subject at a target location an implantable article comprising
a matrix of natural biodegradable material. The article has a
surface, and a structure for treating the condition at the target
location. The method also includes a step of maintaining the
article at the target location for a period of time sufficient to
treat the condition. During the step of maintaining the enzyme is
allowed to contact the article. The structure of the matrix
prevents the enzyme from entering the article, but the enzyme
degrades the article at the surface. The method can be used to
treat a condition such as coronary artery disease.
[0022] In many cases, the article can be completely degraded within
the body. In more specific aspects, in the step of providing, the
matrix comprises cross-linked enzymatically-biodegradable
polymers.
[0023] In some aspects of the invention, the natural biodegradable
material that forms the matrix is a natural biodegradable polymer.
Natural biodegradable polymers include polymer and/or polymer
derivatives that are obtained from natural sources, such as plants
or animals.
[0024] A polymeric matrix can be formed by coupling the
biodegradable polymers together. In some aspects of the invention,
the matrix is formed using natural biodegradable
polysaccharides.
[0025] For example, in preparing the coatings or articles, a
plurality of natural biodegradable polysaccharides are crosslinked
to each other via coupling groups that are pendent from the natural
biodegradable polysaccharide (i.e., one or more coupling groups are
chemically bonded to the polysaccharide). In some aspects, the
coupling group on the natural biodegradable polysaccharide is a
polymerizable group. In a free radical polymerization reaction the
polymerizable group can crosslink natural biodegradable
polysaccharides together in the composition, thereby forming a
natural biodegradable polysaccharide matrix, which can be a portion
of a coating, an in-vivo formed matrix, or the body member of a
medical implant.
[0026] The natural biodegradable polysaccharides described herein
are non-synthetic polysaccharides that can be associated with each
other to form a matrix, which can be used as a coating or as an
article, for example, a medical implant or an in-vivo formed
matrix. The natural biodegradable polysaccharides can also be
enzymatically degraded, but offer the advantage of being generally
non-enzymatically hydrolytically stable. This is particularly
advantageous for bioactive agent delivery, as in some aspects the
invention provides coatings or articles capable of releasing the
bioactive agent under conditions of enzyme-mediated degradation,
but not by diffusion. Therefore, the kinetics of bioactive agent
release from the coatings or articles of the invention are
fundamentally different than those of coatings prepared from
synthetic biodegradable materials, such as poly(lactides).
[0027] Examples of biodegradable articles that can be used to treat
a condition at a target location in a subject include biodegradable
stents.
[0028] Exemplary natural biodegradable polysaccharides include
amylose, maltodextrin, amylopectin, starch, dextran, hyaluronic
acid, heparin, chondroitin sulfate, dermatan sulfate, heparan
sulfate, keratan sulfate, dextran sulfate, pentosan polysulfate,
and chitosan. Preferred polysaccharides are low molecular weight
polymers that have little or no branching, such as those that are
derived from and/or found in starch preparations, for example,
amylose and maltodextrin.
[0029] Because of the particular utility of the amylose and
maltodextrin polymers, in some aspects natural biodegradable
polysaccharides are used that have an average molecular weight of
500,000 Da or less, 250,000 Da or less, 100,000 Da or less, or
50,000 Da or less. In some aspects the natural biodegradable
polysaccharides have an average molecular weight of 500 Da or
greater. In some aspects the natural biodegradable polysaccharides
have an average molecular weight in the range of about 1000 Da to
about 10,000 Da. Natural biodegradable polysaccharides of
particular molecular weights can be obtained commercially or can be
prepared, for example, by acid hydrolysis and/or enzymatic
degradation of a natural biodegradable polysaccharide preparation,
such as starch. The decision of using natural biodegradable
polysaccharides of a particular size range may depend on factors
such as the physical characteristics of the coating composition
(e.g., viscosity), the desired rate of degradation of the coating,
the presence of other optional moieties in the coating composition
(for example, bioactive agents, etc.), etc.
[0030] The natural biodegradable polysaccharides that are used in
accordance with some methods and compositions of the invention are
readily available at a low cost and/or can be prepared easily using
established techniques. This allows for a cost effective method of
coating and fabricating medical articles.
[0031] The use of natural biodegradable material, such as
polysaccharides as exemplified by maltodextrin or amylose, provides
many advantages when used in a coating composition applied to the
surface of a medical device, or for the formation of an article,
such as one that can be used in vivo. Degradation of a natural
biodegradable polysaccharide-containing article, or coating from
the surface of a medical device, can result in the release of, for
example, naturally occurring mono- or disaccharides, such as
glucose, which are common serum components. Furthermore, the use of
natural biodegradable polysaccharides that degrade into common
serum components, such as glucose, can be viewed as more acceptable
than the use of synthetic biodegradable polysaccharides that
degrade into non-natural compounds, or compounds that are found at
very low concentrations in the body.
[0032] In some aspects of the invention, this advantageous feature
is reflected in the use of natural biodegradable materials, such as
natural polysaccharides, as exemplified by amylose and
maltodextrin, and that degrade into products that present little or
no immunogenic or toxic risk to the individual. The invention
provides improved, cost-efficient, natural biodegradable
polysaccharide compositions for articles or coatings that can be
used in a variety of medical treatments.
[0033] Another advantage of the invention is that the natural
biodegradable matrices (such as polysaccharide-based matrices) are
more resistant to hydrolytic degradation than other biodegradable
polymers, such as poly(lactides). As an example, degradation of the
natural biodegradable polysaccharides of the invention are
primarily enzyme-mediated, with minimal or no hydrolysis of the
natural biodegradable polysaccharide occurring when a natural
biodegradable polysaccharide-containing coating is prepared under
ambient conditions. This allows natural biodegradable
polysaccharide-based coatings to remain substantially stable (for
example, resistant to degradation) prior to placing the
coated-article in vivo. For example, a natural biodegradable
polysaccharide coated article can be manipulated in a
non-biological, aqueous-based-medium without risk that the coating
will prematurely degrade due to non-enzyme-mediatated hydrolysis.
Other coatings that are based on biodegradable polymers such as
poly(lactide) or poly(lactide-co-glycolide) are subject to
hydrolysis even at relatively neutral pH ranges (e.g., pH 6.5 to
7.5) and therefore do not offer this advantage.
[0034] Therefore, the invention includes natural
biodegradable-material containing compositions, coatings, articles,
and methods of preparing such that have the advantage of providing
stability in the presence of an aqueous environment.
[0035] In one aspect, the invention provides a shelf-stable
composition for preparing a biodegradable coating, the shelf stable
composition comprising a natural biodegradable polysaccharide
comprising coupling groups. These compositions could be obtained or
prepared, according to the details provided herein, and then stored
for a period of time before the composition is used to form a
biodegradable coating or article, without significant degradation
of the natural biodegradable polysaccharide occurring during
storage. Accordingly, the invention also provides methods for
preparing a biodegradable coating comprising preparing a
biodegradable coating composition comprising a natural
biodegradable polysaccharide comprising coupling group; storing the
coating composition for an amount of time; and then using the
coating composition to prepare a biodegradable coating or a
biodegradable article. In some aspects, the biodegradable article
is formed in situ, for example, by promoting the polymerization of
the natural biodegradable polysaccharide within the body.
Optionally, one or more bioactive agents and/or microparticles can
be added before or after storage of the coating composition.
[0036] In a related aspect, the invention also provides the
advantage of being able to perform methods wherein the natural
biodegradable polysaccharide is subject to exposure to an aqueous
solution without risking significant degradation of the natural
biodegradable polysaccharide. For example, the natural
biodegradable polysaccharide may be contacted with an aqueous
solution in a synthetic or post-synthetic step, including addition
synthesis reactions and purification steps, or a coating that
includes the natural biodegradable polysaccharide can be contacted
with an aqueous solution in, for example, a sterilization step or a
step that involves incorporation of a bioactive agent into the
biodegradable coating.
[0037] In yet another aspect, the invention relates to the
stability of an article or a coating that is formed on an article.
The invention provides a method comprising obtaining an article
formed from, or having a coating comprising, a natural
biodegradable polymer, and then contacting the article with an
aqueous solution for a period of time wherein the article or
coating remains predominantly stable in the solution. The aqueous
solution can be, for example, a storage solution, a solution that
is used to hydrate the surface of the coated device, or an aqueous
sterilization solution.
[0038] Degradation of the coating or article formed of a natural
biodegradable matrix may commence when placed in contact with a
body fluid or tissue, which may include an enzyme capable of
degrading the matrix at its surface. The enzyme can be a natural
biodegradable polysaccharide-degrading enzyme, such as a
carbohydrase.
[0039] The invention also provides a useful way to deliver larger
hydrophilic bioactive agents, such as polypeptides, nucleic acids,
and polysaccharides, as well as viral particles and cells from the
biodegradable article, or biodegradable coating on a surface, such
as a medical device or a coating on a surface thereof.
Comparatively, the use of non-degrading drug delivery matrices may
not be useful for the delivery of these larger bioactive agents if
they are too large to diffuse out of the matrix. However, according
to some aspects of the invention, an article or a coating that
includes a matrix of the natural biodegradable polysaccharide
having a bioactive agent can be placed or formed in the body, and
as the matrix degrades the bioactive agent is gradually released
from the matrix. In one aspect of the invention, the bioactive
agent has a molecular weight of about 10,000 Da or greater.
[0040] In some aspects, the invention provides a drug-releasing
biodegradable article, coating, or composition comprising (i) a
natural biodegradable polysaccharide, preferably selected from
amylose and maltodextrin, comprising an ethylenically unsaturated
group, (ii) an initiator, and (iii) a bioactive agent selected from
the group of polypeptides, polynucleotides, and
polysaccharides.
[0041] In another aspect, a coated surface is prepared on a medical
device, such as a stent or catheter. The methods include disposing
in one or more steps the following reagents on a surface: (a) an
initiator, (b) a natural biodegradable polysaccharide, preferably
selected from amylose and maltodextrin, comprising an ethylenically
unsaturated group, and (c) a bioactive agent. After the components
have been disposed on the surface, the initiator is activated to
crosslink a plurality of natural biodegradable polysaccharides
comprising ethylenically unsaturated groups that are present in the
composition, thereby forming a coating on the surface that includes
the bioactive agent.
[0042] Depending on the application, the initiator can be first
disposed on the surface, followed by disposing the natural
biodegradable polysaccharide and bioactive agent on the layer of
initiator. Alternatively, the initiator, natural biodegradable
polysaccharide, and bioactive agent are mixed and disposed together
on the surface.
[0043] Therefore, in some aspects, the invention provides a method
for delivery of a bioactive agent, or more than one bioactive
agent, to a subject. The method comprises the steps of providing a
coated article to a subject, the coated article having a
biodegradable coating which comprises a plurality of natural
biodegradable polysaccharides associated via coupling groups, and
bioactive agent. The coated article is then exposed to a
carbohydrase to promote the degradation of the coating and release
of the bioactive agent. For example, a biodegradable coating or
article including amylose and/or maltodextrin polymers can be
exposed to an .alpha.-amylase to promote degradation of the coating
and release of the bioactive agent. The step of exposing can be
performed by placing the biodegradable coating or article in a
patient. In the absence of the carbohydrase there is substantially
no release of the bioactive agent. In some aspects the bioactive
agent comprises a polypeptide, such as an antibody or an antibody
fragment.
[0044] In some aspects, the methods of the invention can be used to
prepare coatings wherein an amount of bioactive agent in the range
of 1% to 17% of the total amount of bioactive agent present in the
coating is released from the coating within a period of 2 days, and
coatings wherein an amount of bioactive agent in the range of 1% to
20% of the total amount of bioactive agent present in the coating
is released from the coating within a period of 8 days.
[0045] In other aspects, the bioactive agent is delivered from a
medical implant having a biodegradable body member which comprises
a plurality of natural biodegradable polysaccharide associated via
pendent coupling groups, the body member also including a bioactive
agent. The medical implant is then exposed to a carbohydrase to
promote the degradation of the implant and release of the bioactive
agent.
[0046] In some aspects, the methods of the invention can be used to
prepare medical implants wherein an amount of bioactive agent in
the range of 1% to 17% of the total amount of bioactive agent
present in the medical implant is released within a period of 8
days, medical implants wherein an amount of bioactive agent in the
range of 1% to 41% of the total amount of bioactive agent present
in the medical implant is released within a period of 14 days, and
medical implants wherein an amount of bioactive agent in the range
of 1% to 60% of the total amount of bioactive agent present in the
medical implant is released within a period of 21 days.
[0047] Alternatively a carbohydrase can be administered to a
subject, or the carbohydrase can be provided to a portion of the
article, wherein the carbohydrase is released from the portion and
locally causes the degradation of the coating.
[0048] The coatings can also have favorable bioactive
agent-releasing properties when the coated article has been placed
in the body. In this regard, the present invention provides an
overall improvement in terms of providing coatings for implantable
medical articles. Articles that are fabricated from the
biodegradable polysaccharides can have many of the same beneficial
surface characteristics as provided by the biodegradable
polysaccharide coatings.
[0049] In another aspect of the invention, a natural biodegradable
polysaccharide is modified with a hydrophobic moiety in order to
provide a biodegradable matrix having hydrophobic properties.
Therefore, a biodegradable coating or article can be formed from
natural biodegradable polysaccharide comprising one or more pendent
coupling groups and one or more pendent hydrophobic moieties.
Exemplary hydrophobic moieties include fatty acids and derivatives
thereof, and C.sub.2-C.sub.18 alkyl chains.
[0050] Therefore, in some aspects of the invention, modification of
the natural biodegradable polysaccharide allows for preparation of
coatings or articles that are biodegradable and that can release a
hydrophobic bioactive agent.
[0051] In other aspects, the hydrophobic moiety pendent from the
natural biodegradable has properties of a bioactive agent. Upon
degradation of the matrix, the hydrophobic moiety can be hydrolyzed
from the natural biodegradable polymer and released to provide a
therapeutic effect. One example of a therapeutically useful
hydrophobic moiety is butyric acid.
[0052] In yet another aspect, the invention provides methods and
articles for improving the stability of a bioactive agent that is
delivered from a coating or an article by utilizing a natural
biodegradable non-reducing polysaccharide. The non-reducing
polysaccharide can provide an inert matrix and thereby improve the
stability of sensitive bioactive agents, such as proteins and
enzymes. The article or coating can include a matrix having a
plurality of natural biodegradable non-reducing polysaccharides
along with a bioactive agent, such as a polypeptide. An exemplary
non-reducing polysaccharide comprises polyalditol. Biodegradable
non-reducing polysaccharides can very useful for formulating
coatings or articles that release the bioactive agent over a
prolonged period of time.
[0053] While it is desirable to make coatings or articles that
provide desired properties (for example, bioactive agent release,
wettability, etc.), their actual preparation can be challenging. In
particular, the use of some polysaccharides for preparing coatings
or articles may result in products that are unsuitable for use. For
example, some polysaccharide-based coatings, including those made
from starch-based materials, have the potential to be overly
brittle and inflexible. While these properties may be suitable for
pharmaceutical capsules or tablets they are generally undesirable
as properties for coatings or articles, such as bioactive agent
releasing or sealant coatings, or medical implants.
[0054] Despite this, the present invention demonstrates the
preparation of articles and coatings that include natural
biodegradable polysaccharides that are suitable for in vivo use.
These products display excellent physical characteristics and are
suitable for use in applications wherein a particular function,
such as bioactive agent delivery or a sealant function is desired.
For example, coatings or articles can be prepared having
viscoelastic properties. In one aspect of the invention, the
coating or article has an elastic modulus value in the range of 27
kPa to 30 kPa.
[0055] The coatings of the present invention can have desirable
surface properties that include elasticity and wettability, in
addition to being biodegradable. Also, it has surprisingly been
discovered that the coatings demonstrate excellent lubricity, which
can provide distinct advantages for short term and single use
devices. Therefore, in one aspect, the invention presents a method
for providing lubricity to an article surface comprising the steps
of disposing a composition comprising a plurality of natural
biodegradable polysaccharides comprising pendent coupling groups
and activating the coupling groups to promote association of the
plurality of natural biodegradable polysaccharides and formation of
a lubricious coating on the article surface.
[0056] The coatings can be formed on the surfaces of medical
articles, including those designed for single use or for short-term
use. For example, a lubricious coating can be formed on a
catheter.
[0057] Coatings including natural biodegradable polysaccharides can
be prepared to provide a lubricity of 20 g or less, and can also be
prepared to provide a lubricity of 15 g or less, or 10 g or less,
as based on friction testing. The coatings were also shown to be
highly durable, as lubricity was maintained during multiple cycles
of friction testing. Methods utilizing a photoinitiator have been
shown to provide coatings with both excellent lubricity and
durability.
[0058] In some embodiments of the invention, the methods of
preparing the compositions for fabrication of articles and/or
coated surfaces do not require the use of organic solvents. The use
of organic solvents can be physically hazardous. Use of organic
solvents can potentially destroy the activity of a bioactive agent
that can be optionally included in a natural biodegradable
polysaccharide-based composition.
[0059] Many of the advantageous features of the present natural
biodegradable polysaccharide-containing coatings and articles are
thought to be provided by the starting materials, in particular the
natural biodegradable polysaccharides having pendent coupling
groups. In some aspects the natural biodegradable polysaccharides
have pendent polymerizable groups, such as ethylenically
unsaturated groups. In a preferred aspect, the degradable
polymerizable polymers (macromers) are formed by reacting a natural
biodegradable polysaccharide with a compound comprising an
ethylenically unsaturated group. For example, in some cases, a
natural biodegradable polysaccharide is reacted with a compound
including an ethylenically unsaturated group and an isocyanate
group. In another example of synthesis, a natural biodegradable
polysaccharide is treated with an oxidizing agent to form a
reactive aldehyde species on the polysaccharide and then reacted
with a compound comprising an ethylenically unsaturated group and
an amine group. Polysaccharide macromers were shown to have
excellent matrix forming capabilities.
[0060] Synthesis can be carried out to provide the natural
biodegradable polysaccharide with a desired quantity of pendent
coupling groups. It has been found that use of a natural
biodegradable polysaccharide having a predetermined amount of the
coupling groups allows for the formation of a coating or an article
having desirable physical characteristics (for example, the
coatings are not brittle). Therefore, in some aspects, the
invention provides natural biodegradable polysaccharides having an
amount of pendent coupling groups of about 0.7 .mu.moles of
coupling group per milligram of natural biodegradable
polysaccharide. Preferably the amount of coupling group per natural
biodegradable polysaccharide is in the range of about 0.3
.mu.moles/mg to about 0.7 .mu.moles/mg. For example, amylose or
maltodextrin can be subject to a synthesis reaction with a compound
having an ethylenically unsaturated group to provide an amylose or
maltodextrin macromer having a ethylenically unsaturated group load
level in the range of about 0.3 .mu.moles/mg to about 0.7
.mu.moles/mg.
[0061] In some aspects of the invention an initiator is used to
promote the formation of the natural biodegradable polysaccharide
matrix for article or coating formation. The initiator can be an
independent compound or a pendent chemical group used to activate
the coupling group pendent from the natural biodegradable polymer
and promote coupling of a plurality of natural biodegradable
polymers. When the coupling group pendent from the natural
biodegradable polysaccharide is a polymerizable group, the
initiator can be used in a free radical polymerization reaction to
promote crosslinking of the natural biodegradable polysaccharides
together in the composition.
[0062] Therefore, in one aspect, the invention provides a
biodegradable coating or article composition comprising (i) a
natural biodegradable polysaccharide, preferably selected from
amylose and maltodextrin, comprising a coupling group, (ii) an
initiator, and (iii) a bioactive agent, wherein the coupling group
is able to be activated by the initiator and promote crosslinking
of a plurality of natural biodegradable polysaccharides. In some
aspects of the invention the initiator is independent of the
natural biodegradable polysaccharide and in other aspects the
initiator is pendent from the natural biodegradable polysaccharide.
Preferably, the natural biodegradable polysaccharide comprises an
ethylenically unsaturated group. In some aspects a photoinitiator
is used, such as a photoinitiator that is activated by light
wavelengths having no or a minimal effect on the bioactive agent
present in the composition.
[0063] In another aspect, the initiator includes an oxidizing
agent/reducing agent pair, a "redox pair," to drive polymerization
of the biodegradable polysaccharide. In preparing the biodegradable
coating or article the oxidizing agent and reducing agent are
combined in the presence of the biodegradable polysaccharide. One
benefit of using a redox pair is that, when combined, the oxidizing
agent and reducing agent can provide a particularly robust
initiation system. This is advantageous as it can promote the
formation of a matrix, for example, useful for coating or article
preparation, from biodegradable polysaccharide compositions having
a relatively low viscosity. This can be particularly useful in many
applications, especially when the biodegradable polysaccharide
composition is used for the formation of an in situ polymerized
article. For example, a low viscosity composition can be passed
through a small gauge delivery conduit with relative ease to
provide the composition that can polymerize in situ.
[0064] In some aspects of the invention, the viscosity of the
composition is above about 5 centi Poise (cP), or about 10 cP or
greater. In other aspects of the invention the viscosity of the
composition is between about 5 cP or 10 cP and about 700 cP, and in
some aspects between about 5 cP or 10 cP and about 250 cP. In some
aspects the viscosity of the composition is above about 5 cP or 10
cP and the biodegradable polysaccharides in the composition have an
average molecular weight of 500,000 Da or less, 250,000 Da or less,
100,000 Da or less, or 50,000 Da or less.
[0065] A method for preparing a coating or article can include the
steps of (a) providing a first composition that includes a natural
biodegradable polysaccharide comprising a coupling group and a
first member of a redox pair (for example, the oxidizing agent) and
then (b) mixing the first composition with second composition that
includes a second member of the redox pair (for example, the
reducing agent). In some aspects the second composition includes a
natural biodegradable polysaccharide. For example, the first
composition can include (a) a natural biodegradable polysaccharide
having a coupling group and an oxidizing agent and the second
composition can include a (b) natural biodegradable polysaccharide
having a coupling group and a reducing agent. In some aspects, when
the first composition is combined with the second composition, the
final composition can be about 5 cP or greater.
[0066] The oxidizing agent can be selected from inorganic or
organic oxidizing agents, including enzymes; the reducing agent can
be selected from inorganic or organic reducing agents, including
enzymes. Exemplary oxidizing agents include peroxides, including
hydrogen peroxide, metal oxides, and oxidases, such as glucose
oxidase. Exemplary reducing agents include salts and derivatives of
electropositive elemental metals such as Li, Na, Mg, Fe, Zn, Al,
and reductases. In one aspect, the reducing agent is present in the
composition at a concentration of 2.5 mM or greater when mixed with
the oxidizing agent. Other reagents, such as metal or ammonium
salts of persulfate, can be present in the composition to promote
polymerization of the biodegradable polysaccharide.
[0067] A coating or article formed using redox polymerization can
therefore comprise a plurality of natural biodegradable
polysaccharides associated via polymerized groups, a reduced
oxidizing agent, and an oxidized reducing agent. In one preferred
aspect, the biodegradable coated layer is formed article having a
first coated layer comprising a synthetic polymer.
[0068] The invention also provides alternative methods for
preparing a coated surface or an article that is biodegradable and
that can release a bioactive agent. For example, an alternative
method for forming a coating includes disposing in two or more
steps at least the following reagents on a surface: (a) a natural
biodegradable polysaccharide comprising a first coupling group, (b)
a natural biodegradable polysaccharide comprising a second coupling
group that is reactive with the first coupling group, and (c) a
bioactive agent. According to this method reagents (a) and (b) are
reactive with each other and are disposed separately on the surface
but can individually include reagent (c). For example, reagent (a)
is first disposed on the surface and then a mixture comprising
reagent (b) and (c) is then disposed on reagent (a). Reagent (a)
reacts with (b) to link the natural biodegradable polysaccharides
together to form a coating that includes reagent (c), the bioactive
agent. An article can be formed in a similar manner, for example,
by a method that includes combining (a) a natural biodegradable
polysaccharide comprising a first coupling group with (b) a natural
biodegradable polysaccharide comprising a second coupling group
that is reactive with the first coupling group, and (c) a bioactive
agent. The article can be partially or fully formed when reagent
(a) reacts with (b) to link the natural biodegradable
polysaccharides together to form the article, which includes
reagent (c), the bioactive agent.
[0069] In some aspects, the present invention employs the use of
biodegradable microparticles that include a bioactive agent and a
natural biodegradable polysaccharide, such as amylose and
maltodextrin that have pendent coupling groups. The microparticles
are used in association with the natural biodegradable
polysaccharides to prepare a biodegradable, bioactive
agent-releasing coating for the surface of medical devices.
[0070] According to this aspect of the invention, a medical device
having a coating that includes a crosslinked matrix of natural
biodegradable polysaccharides and biodegradable microparticles
having a bioactive agent can be placed in the body, and as the
biodegradable microparticles degrade the bioactive agent is
gradually released from the coating.
[0071] The natural biodegradable polysaccharide matrix provides the
ability to associate the biodegradable microparticles with the
surface of the coated device. In some arrangements, the
biodegradable microparticles are dispersed in the natural
biodegradable polysaccharide matrix. Such coatings can be formed by
disposing a mixture of (a) biodegradable microparticles having a
bioactive agent and (b) natural biodegradable polysaccharides
having pendent coupling groups, disposing the mixture on a surface,
and then treating the composition to form a coated layer wherein
the biodegradable microparticles are dispersed within the
matrix.
[0072] In other arrangements, the coating is formed by disposing
the biodegradable microparticles independently of the natural
biodegradable polysaccharide having pendent coupling groups. In
these arrangements the biodegradable microparticles can be present
predominantly one face of the layer that is formed from the natural
biodegradable polysaccharide and a microparticle-matrix interface
can be formed.
[0073] The methods include disposing in one or more steps the
following components on a surface: (a) an initiator, (b) a natural
biodegradable polysaccharide, preferably selected from amylose and
maltodextrin, comprising a coupling group, and (c) biodegradable
microparticles comprising a bioactive agent. After the components
have been disposed on the surface, the initiator is activated to
couple a plurality of natural biodegradable polysaccharide polymers
that are present in the composition, thereby forming a natural
biodegradable polysaccharide matrix on the surface that is
associated with the biodegradable microparticles having the
bioactive agent.
[0074] In these aspects, the method includes the steps of (i)
disposing a composition comprising (a) a natural biodegradable
polysaccharide having a coupling group, (b) an initiator, and (c)
biodegradable microparticles comprising a bioactive agent on a
surface; and (ii) activating the initiator to provide a coated
composition having the natural biodegradable polysaccharide and the
biodegradable microparticles having the bioactive agent on the
surface. Alternatively, the initiator can be disposed independently
of the natural biodegradable polysaccharide.
[0075] By including microparticles having a bioactive agent in the
natural biodegradable polysaccharide-containing coating, the
invention also provides a way to effectively and efficiently
prepare a variety of drug-delivery coatings. The use of
microparticles offers the ability to easily prepare coatings having
one or more bioactive agents present in desired amounts in the
coating. Such coatings can be prepared by obtaining biodegradable
microparticles that have a bioactive agent and then forming a
coating that includes the microspheres associated with the natural
biodegradable polysaccharide matrix. In some aspects, different
microparticles having different bioactive agents can be included in
the coating in desired amounts to provide a bioactive
agent-releasing coating that is able to release a desired
combination of bioactive agents in desired amounts. This is a
particular advantage when using bioactive agents that are typically
not compatible in the same composition (for example, bioactive
agents that have different physical properties).
[0076] Microparticles can also be included in articles formed from
the natural biodegradable polysaccharide. For example,
microparticles can be included in an implantable medical article
formed from the natural biodegradable polysaccharides of the
invention, or can be included in an article that is formed in situ.
In these aspects, the presence of biodegradable microparticles in
articles can offer many of the advantages that are offered by the
presence of the microparticles in the coatings.
[0077] In another aspect, the present invention provides
compositions and methods for preparing sealant materials that are
particularly useful in connection with implantable medical articles
having a porous surface, such as grafts, patches, and wound
dressings. In preferred aspects, the inventive compositions can be
used to prepare a sealant coating for implantable medical articles,
particularly implantable medical articles that include a porous
surface.
[0078] The sealant coating can provide a barrier to the movement of
body fluids, such as blood, near the surface of the coated article.
For example, the natural biodegradable polysaccharide-based sealant
coating can provide hemostasis at the article surface by formation
of a tight seal. Gradually, the natural biodegradable
polysaccharide in the sealant coating degrades and a tissue layer
is formed as the sealant coating is replaced by cells and other
factors involved in tissue repair. During the process of
degradation, natural biodegradable polysaccharide degradation
products, such as naturally occurring mono- or disaccharides, for
example, glucose, are released from the sealant coating, which can
be considered an ideal in vivo degradation product because it is
commonly found in the body and may also be utilized by the cells
involved in tissue repair during the degradation/infiltration
process. Gradually, infiltrated tissue growth replaces the function
of the natural biodegradable polysaccharide-containing sealant
coating.
[0079] In some aspects, another particular advantage of the
invention is that release of glucose reduces the likelihood that
the process of natural biodegradable polysaccharide degradation and
tissue infiltration will promote a strong inflammatory response.
This is because the natural biodegradable polysaccharide-based
sealant coating can degrade into materials that are non-antigenic
or that have low antigenicity. Another advantage is that the
degradation products are free of other materials that may cause
disease, such as microbial, viral, or prionic materials potentially
present in animal-derived preparations (such as bovine collagen
preparations).
[0080] The sealant compositions of the invention, which include
natural biodegradable polysaccharides, such as amylose or
maltodextrin polymers, that can be coupled together to form a
matrix (at least a portion of the sealant coating) on the medical
article, can include a bioactive agent, which can be released as
the sealant coating degrades.
[0081] In some aspects, the invention provides a biodegradable
sealant composition comprising (i) a natural biodegradable
polysaccharide comprising a coupling group, and (ii) an initiator,
wherein the coupling group is able to be activated by the initiator
and promote coupling of a plurality of natural biodegradable
polysaccharides. Preferably the natural biodegradable
polysaccharide is a polymer such as amylose or maltodextrin. In
some aspects the sealant composition can also include a bioactive
agent. The initiator can be independent of the natural
biodegradable polysaccharide, pendent from the natural
biodegradable polysaccharide polymer, or both pendent and
independent of the natural biodegradable polysaccharide
polymer.
[0082] Accordingly, the invention also provides methods for
preparing a surface having a sealant coating. The sealant coated
surface is prepared on a medical article or article having a porous
surface. The methods include disposing in one or more steps the
following reagents on a surface: (a) an initiator, and (b) a
natural biodegradable polysaccharide comprising a coupling group.
In some aspects a bioactive agent is also disposed on the surface.
In one preferred aspect, the bioactive agent is a prothrombotic or
procoagulant factor. In these aspects, after the components have
been disposed on the surface, the initiator is activated to couple
the natural biodegradable polysaccharides that are present in the
composition, thereby forming a natural biodegradable polysaccharide
coating on the surface that includes the bioactive agent.
[0083] During the step of activating, the natural biodegradable
polysaccharide is contacted with the initiator and the initiator is
activated to promote the coupling of two or more natural
biodegradable polysaccharides via their coupling groups. In
preferred aspects, the natural biodegradable polysaccharide
includes a polymerizable group, such as an ethylenically
unsaturated group, and initiator is capable of initiating free
radical polymerization of the polymerizable groups.
[0084] The invention also provides alternative methods for
preparing a sealant coating on the surface of an article. The
methods include disposing at least the following reagents on a
surface: (a) a natural biodegradable polysaccharide comprising a
first coupling group and (b) a natural biodegradable polysaccharide
comprising a second coupling group, where in the second coupling
group is reactive with the first coupling group. According to this
method, reagents (a) and (b) are reactive with each other to couple
the natural biodegradable polysaccharide, or (a) and/or (b) can be
treated to be made reactive with each other. In some aspects (a)
and (b) are disposed separately on the surface to form the sealant
coating. The natural biodegradable polysaccharide can be the same
types of polymers of different types of polymers.
[0085] The first coupling group and second coupling group can be a
pair of chemical groups that are reactive with one another,
preferably specifically reactive. The groups can also become
reactive with each other upon addition of a particular agent to the
mixture of natural biodegradable polysaccharide having different
reactive groups.
BRIEF DESCRIPTION OF THE DRAWINGS
[0086] FIG. 1 is a graph of cumulative BSA release from
maltodextrin-acrylate filaments treated with amylase, over a period
of time.
[0087] FIG. 2 is a graph of cumulative BSA release from
maltodextrin-acrylate/photo-PVP-coated PEBAX rod treated with
amylase, over a period of time.
[0088] FIG. 3 is a graph of cumulative absorbance values of active
and total IgG Fab fragment release from maltodextrin-acrylate
filaments treated with amylase, over a period of time.
[0089] FIG. 4 is a graph of cumulative absorbance values of active
and total IgG release from
maltodextrin-acrylate(redox)/photo-PVP-coated stainless steel rods
treated with amylase, over a period of time.
[0090] FIG. 5 is a graph of cumulative absorbance values of active
and total IgG release from
maltodextrin-acrylate(photoinitiation)/photo-PVP-coated stainless
steel rods treated with amylase and percent degradation of the
maltodextrin-acrylate coating, over a period of time.
[0091] FIG. 6 is a graph of cumulative absorbance values of active
and total IgG release from a maltodextrin-acrylate filament treated
with amylase and percent degradation of the filament, over a period
of time.
[0092] FIG. 7 is a graph of modulus of a maltodextrin-acrylate
matrix formed via REDOX polymerization, over a period of time.
[0093] FIG. 8 is a graph of repetitive force testing of
maltodextrin-acrylate coated PEBAX rods versus synthetic
polymer-coated PEBAX rods.
DETAILED DESCRIPTION
[0094] The embodiments of the present invention described herein
are not intended to be exhaustive or to limit the invention to the
precise forms disclosed in the following detailed description.
Rather, the embodiments are chosen and described so that others
skilled in the art can appreciate and understand the principles and
practices of the present invention.
[0095] All publications and patents mentioned herein are hereby
incorporated by reference. The publications and patents disclosed
herein are provided solely for their disclosure. Nothing herein is
to be construed as an admission that the inventors are not entitled
to antedate any publication and/or patent, including any
publication and/or patent cited herein.
[0096] In one aspect, the invention provides methods of preparing
biodegradable coatings. The coatings can release bioactive agents
from the surface of medical devices. The coatings comprise a matrix
of natural biodegradable material. The compositions and methods of
the present invention are particularly useful for coating surfaces
of implantable medical devices, such as stents and catheters, and
that are capable of releasing drugs from the device.
[0097] In another aspect, the invention provides methods of
preparing biodegradable articles, such as medical implants or in
situ formed matrices. The articles comprise a matrix of natural
biodegradable material. The biodegradable articles can also be used
for the release of bioactive agents, and in this manner can
function as bioactive agent-releasing implants or depots. In some
aspects, the biodegradable articles of the invention biodegrade
within a period that is acceptable for the desired application.
[0098] In some aspects, the biodegradable article is a medical
implant that provides mechanical properties at the implantation
site and maintains these mechanical properties until they are no
longer needed. After this period of time has elapsed, the medical
implant is degraded to an extent that the properties are no longer
provided by the medical implant, and the biodegradable components
can be absorbed and/or excreted by the body. In some embodiments,
the medical implant slowly degrades and transfers stress at the
appropriate rate to surrounding tissues as these tissues heal and
can accommodate the stress once borne by the medical device.
[0099] Generally, the biodegradable matrices of the present
invention (whether used alone or in connection with another device)
have a structure that prevents the entrance of an enzyme that is
capable of degrading the matrix, into the matrix. That is, the
structure of the matrix physically excludes higher molecular weight
compounds such as enzymes from entering the matrix. The present
invention shows that matrices treated with a matrix degrading
enzyme (e.g., crosslinked maltodextrin matrices treated with
amylase) exhibited uniform surface degradation. Examination of
these matrices that were subjected to enzymatic degradation for a
period of time revealed a gradual and smooth erosion of the
surface. No bulk erosion of the matrix or irregular surface
patterns (which would otherwise indicate infiltration of the enzyme
into the matrix) were observed. Furthermore, various experiments
show that bioactive agents were released from the matrices at a
zero order rate, further signifying the exclusion properties of the
matrix.
[0100] Enzymes are proteins that generally have a molecular weights
of about 10,000 Da or greater, and more typically 25,000 Da or
greater, or 40,000 Da or greater.
[0101] The matrices of the present invention can exclude and be
degraded by enzymes that degrade naturally occurring materials. One
class of enzymes that can be excluded form the matrix are
carbohydrases, which include polysaccharidases. The carbohydrase
that contacts the coating or article can specifically degrade the
natural biodegradable polysaccharide causing release of the
bioactive agent. Examples of carbohydrases that can specifically
degrade natural biodegradable polysaccharide coatings include
.alpha.-amylases, such as salivary and pancreatic .alpha.-amylases;
disaccharidases, such as maltase, lactase and sucrase;
trisaccharidases; and glucoamylase (amyloglucosidase).
[0102] .alpha.-Amylase extracted from human pancreas and purified
by using ammonium sulfate fractionation has been shown to have a
molecular weight of 53,700 Da according to SDS polyacrylamide gel
electrophoresis (Sky-Peck, H. H. and Thuvasethakul, P. (1977) Human
pancreatic alpha-amylase. I. Purification and characterization.
Ann. Clin. and Lab. Sci. 7:298-309).
[0103] Another class of enzymes that can be excluded from the
matrix is proteases. Proteases can degrade natural biodegradable
materials such as water-soluble proteins. Well known water-soluble
proteins include globular proteins, such as albumin. Albumins can
be degraded by aspartic and cysteine proteinases present in the
body.
[0104] Another class of enzymes that can be excluded from the
matrix is lipases. Lipases are enzymes that catalyze hydrolysis of
fatty acid ester bond in triacylglycerol thus releasing free fatty
acids. The lipase commonly measured in plasma is pancreatic lipase.
Human pancreatic lipase has a molecular weight of about 48,000 Da
(De Caro, A. et al. (1977) Biochim. Biophys. Acta. 490:411-419)
[0105] Another class of enzymes that can be excluded from the
matrix is nucleases. Nucleases include ribonucleases and
deoxyribonucleases. Numerous ribonucleases and deoxyribonucleases
are known which can degrade RNA and DNA molecules in a sequence
specific or non-specific manner. Nucleases, such as human
deoxyribonuclease I have a molecular weight of about 41,000 (Ito,
K. et al. (1984) J. Biochem. (Tokyo). 95:1399-1406) and is present
in human serum (Nadano, D. et al. (1993) Clinical Chem. 39:
448-452).
[0106] In some aspects of the invention, the biodegradable coating
or article includes a natural biodegradable polysaccharide having a
coupling group. Exemplary natural biodegradable polysaccharides
include amylose and maltodextrin. In some aspects, the present
invention provides biodegradable coatings having excellent surface
characteristics and that can provide a suitable vehicle for the
delivery of bioactive agents. These biodegradable coatings can be
disposed on medical devices having a variety of biomaterial
surfaces.
[0107] In some embodiments of the invention, a coating is formed on
a device that includes a biodegradable matrix and biodegradable
microparticles, the biodegradable microparticles including one or
more bioactive agents. The biodegradable material used to form the
matrix includes a natural biodegradable polysaccharide as a
component. In the matrix, natural biodegradable polysaccharides
such as amylose and maltodextrin are coupled to each other and the
biodegradable microparticles are associated with the matrix.
[0108] In yet other embodiments of the invention, a sealant coating
is formed on a device. The sealant coating includes a biodegradable
matrix and optionally one or more bioactive agents, such as
prothrombotic agents.
[0109] The sealant coating of the invention can, at least
initially, provide a barrier on the porous surface that is not
permeable to fluids within the body. Gradually, the sealant coating
degrades and its function is replaced by tissue that infiltrates
the porous surface. Therefore, the sealant coating has particular
properties, such as biodegradability and relative impermeability
(i.e., relative to the degradation of the sealant coating). The
sealant coating can also be compliant and/or conformal, and can
have properties such as flexibility, elasticity, and
bendability.
[0110] As used herein, impermeable, used in relation to the
function of the sealant coating, refers to a significant reduction
in the transmission of bulk liquid or fluids through the substrate
which the sealant coating is associated with. For example, the
sealant coating can be impermeable to the transmission of blood.
The impermeability can be maintained as the natural biodegradable
polysaccharide-based sealant coating degrades, and is replaced by
tissue.
[0111] As referred to herein, a "natural biodegradable
polysaccharide" refers to a non-synthetic polysaccharide that is
capable of being enzymatically degraded but that is generally
non-enzymatically hydrolytically stable. Natural biodegradable
polysaccharides include polysaccharide and/or polysaccharide
derivatives that are obtained from natural sources, such as plants
or animals. Natural biodegradable polysaccharides include any
polysaccharide that has been processed or modified from a natural
biodegradable polysaccharide (for example, maltodextrin is a
natural biodegradable polysaccharide that is processed from
starch). Exemplary natural biodegradable polysaccharides include
hyaluronic acid, starch, dextran, heparin, chondroitin sulfate,
dermatan sulfate, heparan sulfate, keratan sulfate, dextran
sulfate, pentosan polysulfate, and chitosan. Preferred
polysaccharides are low molecular weight polymers that have little
or no branching, such as those that are derived from and/or found
in starch preparations, for example, amylose and maltodextrin.
Therefore, the natural biodegradable polysaccharide can be a
substantially non-branched or non-branched poly(glucopyranose)
polymer.
[0112] Because of the particular utility of the amylose and
maltodextrin polymers, it is preferred that natural biodegradable
polysaccharides having an average molecular weight of 500,000 Da or
less, 250,000 Da or less, 100,000 Da or less, or 50,000 Da or less.
It is also preferred that the natural biodegradable polysaccharides
have an average molecular weight of 500 Da or greater. A
particularly preferred size range for the natural biodegradable
polysaccharides is in the range of about 1000 Da to about 10,000
Da. Natural biodegradable polysaccharides of particular molecular
weights can be obtained commercially or can be prepared. The
decision of using natural biodegradable polysaccharides of a
particular size range may depend on factors such as the physical
characteristics of the coating composition (e.g., viscosity), the
desired rate of degradation of the coating, the presence of other
optional moieties in the coating composition, for example,
bioactive agents, etc.
[0113] As used herein, "amylose" or "amylose polymer" refers to a
linear polymer having repeating glucopyranose units that are joined
by .alpha.-1,4 linkages. Some amylose polymers can have a very
small amount of branching via .alpha.-1,6 linkages (about less than
0.5% of the linkages) but still demonstrate the same physical
properties as linear (unbranched) amylose polymers do. Generally
amylose polymers derived from plant sources have molecular weights
of about 1.times.10.sup.6 Da or less. Amylopectin, comparatively,
is a branched polymer having repeating glucopyranose units that are
joined by .alpha.-1,4 linkages to form linear portions and the
linear portions are linked together via .alpha.-1,6 linkages. The
branch point linkages are generally greater than 1% of the total
linkages and typically 4%-5% of the total linkages. Generally
amylopectin derived from plant sources have molecular weights of
1.times.10.sup.7 Da or greater.
[0114] Amylose can be obtained from, or is present in, a variety of
sources. Typically, amylose is obtained from non-animal sources,
such as plant sources. In some aspects, a purified preparation of
amylose is used as starting material for the preparation of the
amylose polymer having coupling groups. In other aspects, as
starting material, amylose can be used in a mixture that includes
other polysaccharides.
[0115] For example, in some aspects, starch preparations having a
high amylose content, purified amylose, synthetically prepared
amylose, or enriched amylose preparations can be used in the
preparation of amylose having the coupling groups. In starch
sources, amylose is typically present along with amylopectin, which
is a branched polysaccharide. According to the invention, it is
preferred to use coating compositions that include amylose, wherein
the amylose is present in the composition in an amount greater than
amylopectin, if present in the composition. For example, in some
aspects, starch preparations having high amylose content, purified
amylose, synthetically prepared amylose, or enriched amylose
preparations can be used in the preparation of amylose polymer
having the coupling groups. In some embodiments the composition
includes a mixture of polysaccharides including amylose wherein the
amylose content in the mixture of polysaccharides is 50% or
greater, 60% or greater, 70% or greater, 80% or greater, or 85% or
greater by weight. In other embodiments the composition includes a
mixture of polysaccharides including amylose and amylopectin and
wherein the amylopectin content in the mixture of polysaccharides
is 30% or less, or 15% or less. In some cases it may be desirable
to use non-retrograding starches, such as waxy starch, in the
current invention. The amount of amylopectin present in a starch
may also be reduced by treating the starch with amylopectinase,
which cleaves .alpha.-1,6 linkages resulting in the debranching of
amylopectin into amylose.
[0116] In some cases a synthesis reaction can be carried out to
prepare an amylose polymer having pendent coupling groups (for
example, amylose with pendent ethylenically unsaturated groups) and
steps may be performed before, during, and/or after the synthesis
to enrich the amount of amylose, or purify the amylose.
[0117] Amylose of a particular size, or a combination of particular
sizes can be used. The choice of amylose in a particular size range
may depend on the application, for example, the type of surface
coated or the porosity of the surface. In some embodiments amylose
having an average molecular weight of 500,000 Da or less, 250,000
Da or less, 100,000 Da or less, 50,000 Da or less, preferably
greater than 500 Da, or preferably in the range of about 1000 Da to
about 10,000 Da is used. Amylose of particular molecular weights
can be obtained commercially or can be prepared. For example,
synthetic amyloses with average molecular masses of 70, 110, 320,
and 1,000 kDa can be obtained from Nakano Vinegar Co., Ltd. (Aichi,
Japan). The decision of using amylose of a particular size range
may depend on factors such as the physical characteristics of the
coating composition (e.g., viscosity), the desired rate of
degradation of the coating, the presence of other optional moieties
in the coating composition (for example, bioactive agents, etc.),
etc.
[0118] Maltodextrin is typically generated by hydrolyzing a starch
slurry with heat-stable .alpha.-amylase at temperatures at
85-90.degree. C. until the desired degree of hydrolysis is reached
and then inactivating the .alpha.-amylase by a second heat
treatment. The maltodextrin can be purified by filtration and then
spray dried to a final product. Maltodextrins are typically
characterized by their dextrose equivalent (DE) value, which is
related to the degree of hydrolysis defined as: DE=MW
dextrose/number-averaged MW starch hydrolysate.times.100.
[0119] A starch preparation that has been totally hydrolyzed to
dextrose (glucose) has a DE of 100, where as starch has a DE of
about zero. A DE of greater than 0 but less than 100 characterizes
the mean-average molecular weight of a starch hydrolysate, and
maltodextrins are considered to have a DE of less than 20.
Maltodextrins of various molecular weights, for example, in the
range of about 500-5000 Da are commercially available (for example,
from CarboMer, San Diego, Calif.).
[0120] Another contemplated class of natural biodegradable
polysaccharides is natural biodegradable non-reducing
polysaccharides. A non-reducing polysaccharide can provide an inert
matrix thereby improving the stability of sensitive bioactive
agents, such as proteins and enzymes. A non-reducing polysaccharide
refers to a polymer of non-reducing disaccharides (two
monosaccharides linked through their anomeric centers) such as
trehalose (.alpha.-D-glucopyranosyl .alpha.-D-glucopyranoside) and
sucrose (.beta.-D-fructofuranosyl .alpha.-D-glucopyranoside). An
exemplary non-reducing polysaccharide comprises polyalditol which
is available from GPC (Muscatine, Iowa). In another aspect, the
polysaccharide is a glucopyranosyl polymer, such as a polymer that
includes repeating (1.fwdarw.3)O-.beta.-D-glucopyranosyl units.
[0121] In some aspects, the coating compositions can include
natural biodegradable polysaccharides that include chemical
modifications other than the pendent coupling group. To exemplify
this aspect, modified amylose having esterified hydroxyl groups can
be prepared and used in sealant coating compositions in association
with the methods of the invention. Other natural biodegradable
polysaccharides having hydroxyl groups may be modified in the same
manner. These types of modifications can change or improve the
properties of the natural biodegradable polysaccharide making for a
coating composition that is particularly suitable for a desired
application. Many chemically modified amylose polymers, such as
chemically modified starch, have at least been considered
acceptable food additives.
[0122] As used herein, "modified natural biodegradable
polysaccharides" refers to chemical modifications to the natural
biodegradable polysaccharide that are different than those provided
by the coupling group or the initiator group. Modified amylose
polymers having a coupling group (and/or initiator group) can be
used in the compositions and methods of the invention.
[0123] To exemplify this aspect, modified amylose is described. By
chemically modifying the hydroxyl groups of the amylose, the
physical properties of the amylose can be altered. The hydroxyl
groups of amylose allow for extensive hydrogen bonding between
amylose polymers in solution and can result in viscous solutions
that are observed upon heating and then cooling amylose-containing
compositions such as starch in solution (retrograding). The
hydroxyl groups of amylose can be modified to reduce or eliminate
hydrogen bonding between molecules thereby changing the physical
properties of amylose in solution.
[0124] Therefore, in some embodiments the natural biodegradable
polysaccharides, such as amylose, can include one or more
modifications to the hydroxyl groups wherein the modifications are
different than those provided by coupling group. Modifications
include esterification with acetic anhydride (and adipic acid),
succinic anhydride, 1-octenylsuccinic anhydride, phosphoryl
chloride, sodium trimetaphosphate, sodium tripolyphosphate, and
sodium monophosphate; etherification with propylene oxide, acid
modification with hydrochloric acid and sulfuric acids; and
bleaching or oxidation with hydrogen peroxide, peracetic acid,
potassium permanganate, and sodium hypochlorite.
[0125] Examples of modified amylose polymers include carboxymethyl
amylose, carboxyethyl amylose, ethyl amylose, methyl amylose,
hydroxyethyl amylose, hydroxypropyl amylose, acetyl amylose, amino
alkyl amylose, allyl amylose, and oxidized amylose. Other modified
amylose polymers include succinate amylose and oxtenyl succinate
amylose.
[0126] In another aspect of the invention, the natural
biodegradable polysaccharide is modified with a hydrophobic moiety
in order to provide a biodegradable matrix having hydrophobic
properties. Exemplary hydrophobic moieties include those previously
listed, fatty acids and derivatives thereof, and C.sub.2-C.sub.18
alkyl chains. A polysaccharide, such as amylose or maltodextrin,
can be modified with a compound having a hydrophobic moiety, such
as a fatty acid anhydride. The hydroxyl group of a polysaccharide
can also cause the ring opening of lactones to provide pendent
open-chain hydroxy esters.
[0127] In some aspects, the hydrophobic moiety pendent from the
natural biodegradable has properties of a bioactive agent. The
hydrophobic moiety can be hydrolyzed from the natural biodegradable
polymer and released from the matrix to provide a therapeutic
effect. One example of a therapeutically useful hydrophobic moiety
is butyric acid, which has been shown to elicit tumor cell
differentiation and apoptosis, and is thought to be useful for the
treatment of cancer and other blood diseases. The hydrophobic
moiety that provides a therapeutic effect can also be a natural
compound (such as butyric acid). Therefore, degradation of the
matrix having a coupled therapeutic agent can result in all natural
degradation products.
[0128] According to the invention, a natural biodegradable
polysaccharide that includes a coupling group is used to form an
article or a coating on the surface of a medical article. Other
polysaccharides can also be present in the coating composition. For
example, the two or more natural biodegradable polysaccharides are
used to form an article or a coating on the surface of a medical
article. Examples include amylose and one or more other natural
biodegradable polysaccharide(s), and maltodextrin and one or more
other natural biodegradable polysaccharide(s); in one aspect the
composition includes a mixture of amylose and maltodextrin,
optionally with another natural biodegradable polysaccharide.
[0129] In one preferred embodiment, amylose or maltodextrin is the
primary polysaccharide. In some embodiments, the composition
includes a mixture of polysaccharides including amylose or
maltodextrin and the amylose or maltodextrin content in the mixture
of polysaccharides is 50% or greater, 60% or greater, 70% or
greater, 80% or greater, or 85% or greater by weight.
[0130] Purified or enriched amylose preparations can be obtained
commercially or can be prepared using standard biochemical
techniques such as chromatography. In some aspects, high-amylose
cornstarch can be used.
[0131] As used herein, "coupling group" can include (1) a chemical
group that is able to form a reactive species that can react with
the same or similar chemical group to form a bond that is able to
couple the natural biodegradable polysaccharides together (for
example, wherein the formation of a reactive species can be
promoted by an initiator); or (2) a pair of two different chemical
groups that are able to specifically react to form a bond that is
able to couple the natural biodegradable polysaccharides together.
The coupling group can be attached to any suitable natural
biodegradable polysaccharide, including the amylose and
maltodextrin polymers as exemplified herein.
[0132] Contemplated reactive pairs include Reactive Group A and
corresponding Reactive Group B as shown in the Table 1 below. For
the preparation of a coating composition, a reactive group from
group A can be selected and coupled to a first set of natural
biodegradable polysaccharides and a corresponding reactive group B
can be selected and coupled to a second set of natural
biodegradable polysaccharides. Reactive groups A and B can
represent first and second coupling groups, respectively. At least
one and preferably two, or more than two reactive groups are
coupled to an individual natural biodegradable polysaccharide
polymer. The first and second sets of natural biodegradable
polysaccharides can be combined and reacted, for example,
thermochemically, if necessary, to promote the coupling of natural
biodegradable polysaccharides and the formation of a natural
biodegradable polysaccharide matrix.
TABLE-US-00001 TABLE 1 Reactive group A Reactive group B amine
hydroxyl, sulfhydryl N-oxysuccinimide ("NOS") amine Aldehyde amine
Isothiocyanate amine, sulfhydryl Bromoacetyl amine, sulfhydryl
Chloroacetyl amine, sulfhydryl Iodoacetyl amine, hydroxyl Anhydride
aldehyde Hydrazide amine, hydroxyl, carboxylic acid Isocyanate
amine, sulfhydryl Maleimide sulfhydryl Vinylsulfone Amine also
includes hydrazide (R--NH--NH.sub.2)
[0133] For example, a suitable coupling pair would be a natural
biodegradable polysaccharide having an electrophilic group and a
natural biodegradable polysaccharide having a nucleophilic group.
An example of a suitable electrophilic-nucleophilic pair is
N-hydroxysuccinimide-amine pair, respectively. Another suitable
pair would be an oxirane-amine pair.
[0134] In some aspects, the natural biodegradable polysaccharides
of the invention include at least one, and more typically more than
one, coupling group per natural biodegradable polysaccharide,
allowing for a plurality of natural biodegradable polysaccharides
to be coupled in linear and/or branched manner. In some preferred
embodiments, the natural biodegradable polysaccharide includes two
or more pendent coupling groups.
[0135] In some aspects, the coupling group on the natural
biodegradable polysaccharide is a polymerizable group. In a free
radical polymerization reaction the polymerizable group can couple
natural biodegradable polysaccharides together in the composition,
thereby forming a biodegradable natural biodegradable
polysaccharide matrix.
[0136] A preferred polymerizable group is an ethylenically
unsaturated group. Suitable ethylenically unsaturated groups
include vinyl groups, acrylate groups, methacrylate groups,
ethacrylate groups, 2-phenyl acrylate groups, acrylamide groups,
methacrylamide groups, itaconate groups, and styrene groups.
Combinations of different ethylenically unsaturated groups can be
present on a natural biodegradable polysaccharide, such as amylose
or maltodextrin.
[0137] In preparing the natural biodegradable polysaccharide having
pendent coupling groups any suitable synthesis procedure can be
used. Suitable synthetic schemes typically involve reaction of, for
example, hydroxyl groups on the natural biodegradable
polysaccharide, such as amylose or maltodextrin. Synthetic
procedures can be modified to produce a desired number of coupling
groups pendent from the natural biodegradable polysaccharide
backbone. For example, the hydroxyl groups can be reacted with a
coupling group-containing compound or can be modified to be
reactive with a coupling group-containing compound. The number
and/or density of acrylate groups can be controlled using the
present method, for example, by controlling the relative
concentration of reactive moiety to saccharide group content.
[0138] In some modes of practice, the biodegradable polysaccharides
have an amount of pendent coupling groups of about 0.7 .mu.moles of
coupling group per milligram of natural biodegradable
polysaccharide. In a preferred aspect, the amount of coupling group
per natural biodegradable polysaccharide is in the range of about
0.3 .mu.moles/mg to about 0.7 .mu.moles/mg. For example, amylose or
maltodextrin can be reacted with an acrylate groups-containing
compound to provide an amylose or maltodextrin macromer having a
acrylate group load level in the range of about 0.3 .mu.moles/mg to
about 0.7 .mu.moles/mg.
[0139] As used herein, an "initiator" refers to a compound, or more
than one compound, that is capable of promoting the formation of a
reactive species from the coupling group. For example, the
initiator can promote a free radical reaction of natural
biodegradable polysaccharide having a coupling group. In one
embodiment the initiator is a photoreactive group (photoinitiator)
that is activated by radiation. In some embodiments, the initiator
can be an "initiator polymer" that includes a polymer having a
backbone and one or more initiator groups pendent from the backbone
of the polymer.
[0140] In some aspects the initiator is a compound that is light
sensitive and that can be activated to promote the coupling of the
amylose polymer via a free radical polymerization reaction. These
types of initiators are referred to herein as "photoinitiators." In
some aspects it is preferred to use photoinitiators that are
activated by light wavelengths that have no or a minimal effect on
a bioactive agent if present in the composition. A photoinitiator
can be present in a sealant composition independent of the amylose
polymer or pendent from the amylose polymer.
[0141] In some embodiments, photoinitiation occurs using groups
that promote an intra- or intermolecular hydrogen abstraction
reaction. This initiation system can be used without additional
energy transfer acceptor molecules and utilizing nonspecific
hydrogen abstraction, but is more commonly used with an energy
transfer acceptor, typically a tertiary amine, which results in the
formation of both aminoalkyl radicals and ketyl radicals. Examples
of molecules exhibiting hydrogen abstraction reactivity and useful
in a polymeric initiating system, include analogs of benzophenone,
thioxanthone, and camphorquinone.
[0142] In some preferred embodiments the photoinitiator includes
one or more charged groups. The presence of charged groups can
increase the solubility of the photoinitiator (which can contain
photoreactive groups such as aryl ketones) in an aqueous system and
therefore provide for an improved coating composition. Suitable
charged groups include, for example, salts of organic acids, such
as sulfonate, phosphonate, carboxylate, and the like, and onium
groups, such as quaternary ammonium, sulfonium, phosphonium,
protonated amine, and the like. According to this embodiment, a
suitable photoinitiator can include, for example, one or more aryl
ketone photogroups selected from acetophenone, benzophenone,
anthraquinone, anthrone, anthrone-like heterocycles, and
derivatives thereof; and one or more charged groups, for example,
as described herein. Examples of these types of water-soluble
photoinitiators have been described in U.S. Pat. No. 6,077,698.
[0143] In some aspects the photoinitiator is a compound that is
activated by long-wavelength ultraviolet (UV) and visible light
wavelengths. For example, the initiator includes a photoreducible
or photo-oxidizable dye. Photoreducible dyes can also be used in
conjunction with a compound such as a tertiary amine. The tertiary
amine intercepts the induced triplet producing the radical anion of
the dye and the radical cation of the tertiary amine. Examples of
molecules exhibiting photosensitization reactivity and useful as an
initiator include acridine orange, camphorquinone, ethyl eosin,
eosin Y, erythrosine, fluorescein, methylene green, methylene blue,
phloxime, riboflavin, rose bengal, thionine, and xanthine dyes. Use
of these types of photoinitiators can be particularly advantageous
when a light-sensitive bioactive agent is included in the sealant
coating.
[0144] Therefore, in yet another aspect, the invention provides a
coating composition comprising (i) a natural biodegradable
polysaccharide comprising an ethylenically unsaturated group (ii) a
photoinitiator selected from the group consisting of acridine
orange, camphorquinone, ethyl eosin, eosin Y, erythrosine,
fluorescein, methylene green, methylene blue, phloxime, riboflavin,
rose bengal, thionine, and xanthine dyes, and (iii) a bioactive
agent.
[0145] Thermally reactive initiators can also be used to promote
the polymerization of natural biodegradable polymers having pendent
coupling groups. Examples of thermally reactive initiators include
4,4' azobis(4-cyanopentanoic acid),
2,2-azobis[2-(2-imidazolin-2-yl) propane] dihydrochloride, and
analogs of benzoyl peroxide. Redox initiators can also be used to
promote the polymerization of the natural biodegradable polymers
having pendent coupling groups. In general, combinations of organic
and inorganic oxidizers, and organic and inorganic reducing agents
are used to generate radicals for polymerization. A description of
redox initiation can be found in Principles of Polymerization,
2.sup.nd Edition, Odian G., John Wiley and Sons, pgs 201-204,
(1981).
[0146] In some cases, the initiator can be included in a base
coating and the natural biodegradable polysaccharide or composition
that includes the natural biodegradable polysaccharide can be
disposed on the base coating. For example, a coated layer that
includes the natural biodegradable polysaccharide can be formed on
a coated layer that includes a synthetic polymer. The synthetic
polymer can be a hydrophilic polymer such as
poly(vinylpyrrolidone), poly(acrylamide), or copolymers thereof. In
some aspects the synthetic polymer is formed using photoreactive
groups, such as photoreactive groups that are pendent from the
synthetic polymer, which can be used to covalently bond the
synthetic polymer to a surface of the article.
[0147] In some aspects the polymerization initiator is a polymer
that includes an initiator group (herein referred to as an
"initiator polymer"). The polymeric portion of the initiator
polymer can be obtained or prepared to have particular properties
or features that are desirable for use with a coating composition,
such as a sealant coating composition. For example, the polymeric
portion of the initiator polymer can have hydrophilic or amphoteric
properties, it can include pendent charged groups, or it can have
groups that allow it to interact with a particular surface (this
can depend on the type of surface to be coated). Optionally, or
additionally, the polymer can change or improve the properties of
the coating that is formed by the amylose polymer having coupling
groups. For example, the initiator polymer can change the
elasticity, flexibility, wettability, or softness (or combinations
thereof) of the coating formed on the surface. Certain polymers, as
described herein, are useful as plasticizing agents for coatings
that include natural biodegradable polysaccharides. Initiator
groups can be added to these plasticizing polymers and used in the
compositions and methods of the invention.
[0148] For example, in some aspects an initiator can be pendent
from a natural biodegradable polysaccharide. Therefore, the natural
biodegradable polysaccharide is able to promote activation of
polymerizable groups that are pendent from other natural
biodegradable polysaccharides and promote the formation of a
natural biodegradable polysaccharide matrix.
[0149] In other cases, the polymeric portion of the initiator
polymer can include, for example, acrylamide and methacrylamide
monomeric units, or derivatives thereof. In some embodiments, the
coating composition includes an initiator polymer having a
photoreactive group and a polymeric portion selected from the group
of acrylamide and methacrylamide polymers and copolymers.
[0150] In some aspects, the initiator includes an oxidizing
agent/reducing agent pair, a "redox pair," to drive polymerization
of the biodegradable polysaccharide. In this case, polymerization
of the biodegradable polysaccharide is carried out upon combining
one or more oxidizing agents with one or more reducing agents.
Other compounds can be included in the composition to promote
polymerization of the biodegradable polysaccharides.
[0151] When combined, the oxidizing agent and reducing agent can
provide a particularly robust initiation system and can drive the
formation of a polymerized matrix of polysaccharides from a
composition having a low viscosity. A polysaccharide composition
with a low viscosity may be due to a low concentration of
polysaccharide in the composition, a polysaccharide having a low
average molecular weight, or combinations thereof. Matrix formation
from a polysaccharide composition having a low viscosity is
particularly advantageous in many applications, especially for in
situ polymerization. In some aspects of the invention, a low
viscosity polysaccharide composition is passed through a small
gauge delivery conduit, such as a needle, wherein the redox pair
causes the polymerization of the polysaccharides in situ.
[0152] In some aspects of the invention, the viscosity of the
composition is above about 5 cP, or about 10 cP or greater. In
other aspects of the invention the viscosity of the composition is
between about 5 cP or 10 cP and about 700 cP, or between about 5 cP
or 10 cP and about 250 cP.
[0153] In order to promote polymerization of the biodegradable
polysaccharides in a composition to form a matrix, the oxidizing
agent is added to the reducing agent in the presence of the one or
more biodegradable polysaccharides. For example, a composition
including a biodegradable polysaccharide and a reducing agent is
added to a composition including an oxidizing agent, or a
composition including a biodegradable polysaccharide and an
oxidizing agent is added to a composition containing a reducing
agent. One desirable method of preparing a matrix is to combine a
composition including a biodegradable polysaccharide and an
oxidizing agent with a composition including a biodegradable
polysaccharide and a reducing agent. For purposes of describing
this method, the terms "first composition" and "second composition"
can be used.
[0154] The viscosities of biodegradable polysaccharide in the first
and second compositions can be the same or can be different.
Generally, though, it has been observed that good mixing and
subsequent matrix formation is obtained when the compositions have
the same or similar viscosities. In this regard, if the same
biodegradable polymer is used in the first and second compositions,
the concentration of the biodegradable polymer may be the same or
different.
[0155] The oxidizing agent can be selected from inorganic or
organic oxidizing agents, including enzymes; the reducing agent can
be selected from inorganic or organic reducing agents, including
enzymes. Exemplary oxidizing agents include peroxides, including
hydrogen peroxide, metal oxides, and oxidases, including glucose
oxidase. Exemplary reducing agents include salts and derivatives of
electropositive elemental metals such as Li, Na, Mg, Fe, Zn, Al,
and reductases. In one mode of practice, the reducing agent is
present at a concentration of about 2.5 mM or greater when the
reducing agent is mixed with the oxidizing agent. Prior to mixing,
the reducing agent can be present in a composition at a
concentration of, for example, 5 mM or greater.
[0156] Other reagents can be present in the composition to promote
polymerization of the biodegradable polysaccharide. Other
polymerization promoting compounds can be included in the
composition, such as metal or ammonium salts of persulfate.
[0157] Optionally, the compositions and methods of the invention
can include polymerization accelerants that can improve the
efficiency of polymerization. Examples of useful accelerants
include N-vinyl compounds, particularly N-vinyl pyrrolidone and
N-vinyl caprolactam. Such accelerants can be used, for instance, at
a concentration of between about 0.01% and about 5%, and preferably
between about 0.05% and about 0.5%, by weight, based on the volume
of the coating composition.
[0158] In some aspects, an aqueous composition that includes the
natural biodegradable polysaccharide, such as amylose or
maltodextrin having pendent coupling groups, and a bioactive agent
is obtained and used in the method of coating a surface. In another
aspect, an aqueous composition is used to form an article. This
composition can be prepared by mixing a bioactive agent, such as a
water-soluble small molecule, a protein, or a nucleic acid, with
the natural biodegradable polysaccharide.
[0159] According to the invention, the natural biodegradable
polysaccharide that includes a coupling group is used to form a
coating on the surface of a medical device or to form an article.
Other polysaccharides can also be present in the coating
composition. For example, the coating can include two different
natural biodegradable polysaccharides, or more than two different
natural biodegradable polysaccharides. For example, in some cases
the natural biodegradable polysaccharide (such as amylose or
maltodextrin) can be present in the coating or article composition
along with another biodegradable polymer (i.e., a secondary
polymer), or more than one other biodegradable polymer. An
additional polymer or polymers can be used to alter the properties
of the matrix, or serve as bulk polymers to alter the volume of the
matrix. For example, other biodegradable polysaccharides can be
used in combination with the amylose polymer. These include
hyaluronic acid, dextran, starch, amylose (for example,
non-derivitized), amylopectin, cellulose, xanthan, pullulan,
chitosan, pectin, inulin, alginates, and heparin.
[0160] In some aspects of the invention, a composition is disposed
on a surface that includes at least the natural biodegradable
polysaccharide, such as amylose or maltodextrin having a coupling
group and a bioactive agent. In some embodiments the composition
includes the natural biodegradable polysaccharide, a bioactive
agent, and an initiator. In other embodiments, a coating is formed
by disposing the natural biodegradable polysaccharide and disposing
the biodegradable microparticles on a surface. In some embodiments
a composition containing both the natural biodegradable
polysaccharide and the biodegradable microparticles having the
bioactive agent are disposed on a surface. In yet other embodiments
of the invention, a sealant composition that includes at least the
natural biodegradable polysaccharide having a coupling group is
disposed on a porous surface.
[0161] The concentration of the natural biodegradable
polysaccharide in the composition can be chosen to provide a
coating or an article having a desired density of crosslinked
natural biodegradable polysaccharide. In some embodiments, the
concentration of natural biodegradable polysaccharide in the
composition can depend on the type or nature of the bioactive agent
that is included in the composition. In some embodiments the
natural biodegradable polysaccharide having the coupling groups is
present in the coating composition at a concentration in the range
of 5-100% (w/v), and 5-50%, and in more specific embodiments in the
range of 10-20% and in other embodiments in the range of 20-50%
(w/v).
[0162] For example, in forming a medical implant, the concentration
of the natural biodegradable polysaccharide may be higher to
provide a more structurally rigid implant.
[0163] Other polymers or non-polymeric compounds can be included in
the composition that can change or improve the properties of the
coating or article that is formed by the natural biodegradable
coating having coupling groups in order to change the elasticity,
flexibility, wettability, or adherent properties, (or combinations
thereof) of the coating formed on the surface.
[0164] For example, in order to improve the properties of a
coating, such as a sealant coating when formed, it is possible to
include in the mixture one or a combination of plasticizing agents.
Suitable plasticizing agents include glycerol, diethylene glycol,
sorbitol, sorbitol esters, maltitol, sucrose, fructose, invert
sugars, corn syrup, and mixtures thereof. The amount and type of
plasticizing agents can be readily determined using known standards
and techniques.
[0165] Compositions of this invention can be used to coat the
surface of a variety of implantable devices. The coating of natural
biodegradable polysaccharide (with or without bioactive agent) can
be applied to a medical device using standard techniques to cover
the entire surface of the device, or a portion of the device
surface.
[0166] The medical articles on which the biodegradable coating can
be formed can be fabricated from any suitable biomaterial or
combinations of biomaterials. Preferred biomaterials include those
formed of synthetic polymers, including oligomers, homopolymers,
and copolymers resulting from either addition or condensation
polymerizations.
[0167] Examples of suitable addition polymers include, but are not
limited to, acrylics such as those polymerized from methyl
acrylate, methyl methacrylate, hydroxyethyl methacrylate,
hydroxyethyl acrylate, acrylic acid, methacrylic acid, glyceryl
acrylate, glyceryl methacrylate, methacrylamide, and acrylamide;
vinyls such as ethylene, propylene, vinyl chloride, vinyl acetate,
vinyl pyrrolidone, and vinylidene difluoride.
[0168] Examples of condensation polymers include, but are not
limited to, nylons such as polycaprolactam, polylauryl lactam,
polyhexamethylene adipamide, and polyhexamethylene dodecanediamide,
and also polyurethanes, polycarbonates, polyamides, polysulfones,
poly(ethylene terephthalate), polylactic acid, polyglycolic acid,
polydimethylsiloxanes, and polyetherketone.
[0169] Other suitable biomaterials include metals, metal alloys,
and ceramics. The metals and metal alloys include, but are not
limited to, titanium, Nitinol, stainless steel, tantalum, and
cobalt chromium. A second class of metals includes the noble metals
such as gold, silver, copper, and platinum uridium. The ceramics
include, but are not limited to, silicon nitride, silicon carbide,
zirconia, and alumina, as well as glass, silica, and sapphire.
Combinations of ceramics and metals are another class of
biomaterials.
[0170] Certain natural materials are also suitable biomaterials,
including human tissue such as bone, cartilage, skin and teeth; and
other organic materials such as wood, cellulose, compressed carbon,
and rubber.
[0171] The surface of such biomaterials can be pretreated (for
example, with a Parylene coating composition) in order to alter the
surface properties of the biomaterial, when desired.
[0172] The biomaterials as described herein can be used to
fabricate a variety of implantable devices on which the
biodegradable coating can be formed. The medical device can be any
device that is introduced temporarily or permanently into a mammal
for the prophylaxis or treatment of a medical condition. These
devices include any that are introduced subcutaneously,
percutaneously or surgically to rest within an organ, tissue, or
lumen of an organ, such as arteries, veins, ventricles or atria of
the heart. The device can be a biostable device, a partially
degradable device, or a completely degradable device (for example,
stents can be fabricated from biodegradable polymeric
materials).
[0173] The natural biodegradable polysaccharide coating (in some
embodiments including biodegradable microparticles) can be formed
on the surface of virtually any implantable device. Exemplary
implantable devices include but are not limited to drug-delivering
vascular stents; other vascular devices (e.g., grafts, catheters,
valves, artificial hearts, heart assist devices); implantable
defibrillators; blood oxygenator devices; surgical devices;
tissue-related materials; membranes; cell culture devices;
chromatographic support materials; biosensors; shunts for
hydrocephalus; wound management devices; endoscopic devices;
infection control devices; orthopedic devices; dental devices,
urological devices; colostomy bag attachment devices; ophthalmic
devices; glaucoma drain shunts; synthetic prostheses; intraocular
lenses; respiratory, peripheral cardiovascular, spinal,
neurological, dental, and ear/nose/throat devices (e.g., ear
drainage tubes); renal devices; and dialysis articles (e.g.,
tubing, membranes, grafts).
[0174] Other contemplated devices include self-expanding stents
(e.g., made from nitinol), balloon-expanded stents (e.g., prepared
from stainless steel), degradable coronary stents, non-degradable
coronary stents, peripheral coronary stents, urinary catheters
(e.g., surface-coated with antimicrobial agents), penile implants,
sphincter devices, urethral devices, bladder devices, renal
devices, vascular implants and grafts, intravenous catheters (e.g.,
treated with antithrombotic agents), small diameter grafts,
artificial lung catheters, electrophysiology catheters, anastomosis
devices, vertebral disks, bone pins, suture anchors, hemostatic
barriers, clamps, surgical staples/sutures/screws/plates/clips,
atrial septal defect closures, electro-stimulation leads for
cardiac rhythm management (e.g., pacer leads), glucose sensors
(long-term and short-term); blood pressure and stent graft
catheters, blood oxygenator tubing, blood oxygenator membranes,
blood bags, birth control devices, breast implants, benign
prostatic hyperplasia and prostate cancer implants, bone
repair/augmentation devices, breast implants, cartilage repair
devices, orthopedic joint implants, orthopedic fracture repairs,
tissue adhesives, tissue sealants, tissue scaffolds, cerebral
spinal fluid (CSF) shunts, dental implants, dental fracture repair
devices, implanted drug infusion tubes, intravitreal drug delivery
devices, nerve regeneration conduits, oncological implants,
electrostimulation leads, pain management implants,
spinal/orthopedic repair devices, wound dressings, embolic
protection filters, abdominal aortic aneurysm grafts, heart valves
(e.g., mechanical, polymeric, tissue, percutaneous, carbon, sewing
cuff), valve annuloplasty devices, mitral valve repair devices,
vascular intervention devices, left ventricle assist devices, neuro
aneurysm treatment coils, neurological catheters, left atrial
appendage filters, central venous access catheters, hemodialysis
devices, catheter cuffs, anastomotic closures, vascular access
catheters, cardiac sensors, uterine bleeding patches, urological
catheters/stents/implants, in vitro diagnostics, aneurysm exclusion
devices, neuropatches, Vena cava filters, urinary dialators,
endoscopic surgical tissue extractors, atherectomy catheters, clot
extraction catheters, percutaneous transluminal angioplasty (PTA)
catheters, percutaneous transluminal coronary angioplasty (PTCA)
catheters, stylets (vascular and non-vascular), coronary
guidewires, drug infusion catheters, esophageal stents, circulatory
support systems, angiographic catheters, transition sheaths and
dialators, coronary and peripheral guidewires, hemodialysis
catheters, neurovascular balloon catheters, tympanostomy vent
tubes, cerebro-spinal fluid shunts, defibrillator leads,
percutaneous closure devices, drainage tubes, thoracic cavity
suction drainage catheters, electrophysiology catheters, stroke
therapy catheters, abscess drainage catheters, biliary drainage
products, dialysis catheters, central venous access catheters, and
parental feeding catheters.
[0175] The compositions are particularly useful for forming
biodegradable coatings on the surface of devices that will come in
contact with aqueous systems. The body fluids typically have
enzymes that allow for the degradation of the natural biodegradable
polysaccharide-based coating. The aqueous system (such as bodily
fluids) allows for the degradation of the biodegradable coating and
release of the bioactive agent from the device. In some cases,
depending on the bioactive agent and the matrix, the bioactive
agent can diffuse out of the matrix. For example, it has been
demonstrated that a loosely formed matrix may allow some diffusion
of bioactive agents, particularly smaller bioactive agents. More
desirably, well-formed matrices having signification polysaccharide
association via coupling groups are able to retain bioactive
agents. Release of bioactive agents from these matrices is mediated
by enzymatic degradation.
[0176] The coatings can also be formed on a biological article. A
"biological article" refers to any sort of non-synthetic
biologically-based article such as a cell or a portion of a cell, a
group of cells, tissue, or an organ or a portion of a organ. The
present reagents can be used in methods for encapsulating cellular
material.
[0177] In some aspects of the invention, a sealant coating is
provided on a porous surface of a medical article. The medical
article can be any article that is introduced into a mammal for the
prophylaxis or treatment of a medical condition, wherein the
medical article include a sealant coating (at least initially) and
has a sealant function. The medical article having the sealant
coating can provide one or more functions, including providing a
barrier to the movement of body fluids, such as blood.
[0178] The sealant coatings can be formed on the surface of
articles that have a porous structure wherein it is desired to seal
the porous structure, providing a barrier to the movement of body
fluids. In many cases it is desirable to form these artificial
barriers to ensure that the implanted article functions as it is
intended to in the body. Gradually, however, it is desired to allow
the body to maintain the function of the sealant coating by
replacing the sealant barrier materials with natural materials from
the body.
[0179] The sealant composition can be prepared and/or applied in
such a manner as to fill the pores on the surface of the article
with the sealant material. This can be achieved by, for example,
controlling factors such as the viscosity of the coating
composition and the coupling of the natural biodegradable
polysaccharides during formation of the coating.
[0180] An article having a "porous surface" refers to any article
having a surface with pores on which a natural biodegradable
polysaccharide-based sealant coating can be formed. The pores are
preferably of a physical dimension that permits in-growth of tissue
into the pores as the sealant coating degrades. The porous surface
can be associated with a non-porous surface, such as a scaffold
that can provide support to the porous surface.
[0181] The medical article can include porous surfaces that can be
provided with a sealant coating and non-porous surfaces that are
not coated with the sealant coating, optionally coated with the
sealant coating, or coated with a material that is different than
the sealant coating. All or a portion of the porous surfaces can be
coated with the sealant coating. In some cases a sealant material
that is different than the natural biodegradable
polysaccharide-based sealant material can be used in conjunction
with the natural biodegradable polysaccharide-based sealant
material.
[0182] For articles that have interior and exterior porous
surfaces, either the interior or the exterior portions can be
coated, or portions of the interior and/or exterior can be coated.
The portion or portions of the article that are coated can depend
on a particular desired application or function of the coated
article. For example, in some cases it may be desirable to have a
difference in the flow of fluids, such as blood, through porous
portions of the medical article. Also, tissue in-growth on selected
portions of the article can also be promoted by depositing the
sealant coating at desired locations.
[0183] The porous surface of the article can also include a
material that is thrombogenic and/or presents surface stasis areas
(regions of minimized or no blood flow). Depending on the
application, a surface having a desired degree of porosity is
obtained. The surface will have a degree of porosity sufficient for
proper in-growth of cells and tissue forming factors. Upon tissue
in-growth, the surface can provide a barrier that is fluid
impermeable.
[0184] In many cases the porous surface of the article is a fabric
or has fabric-like qualities. The porous surface can be formed from
textiles, which include woven materials, knitted materials, and
braided materials. Particularly useful textile materials are woven
materials which can be formed using any suitable weave pattern
known in the art.
[0185] The porous surface can be that of a graft, sheath, cover,
patch, sleeve, wrap, casing, and the like. These types of articles
can function as the medical article itself or be used in
conjunction with another part of a medical article (examples of
which are described herein).
[0186] The porous surface can include any suitable type of
biomaterial. Useful biomaterials can be woven into fibers for the
preparation of fabrics as described herein. Useful materials
include synthetic addition or condensation polymers such as
polyesters, polypropylenes, polyethylenes, polyurethanes, and
polytetrafluoroethylenes. Polyethylene terephthalate (PET) is a
commonly used polymer in fabrics. Blends of these polymers can also
be utilized in the preparation of fibers, such as monofilament or
multi-filament fibers, for the construction of fabrics. Commonly
used fabrics include those such as nylon, velour, and
DACRON.TM..
[0187] The fabrics can optionally include stiffening materials to
improve the physical properties of the article, for example, to
improve the strength of a graft. Such materials can improve the
function of an implanted article. For example, strengthening
materials can improve the patency of the graft.
[0188] Porous surfaces can also be formed by dipping mandrels in
these types of polymers.
[0189] Other particular contemplated porous surfaces include those
of cardiac patches. These can be used to decrease suture line
bleeding associated with cardiovascular reconstructions. The
patches can be used to seal around the penetrating suture. Common
materials used in cardiac patches include PTFE and DACRON.TM..
[0190] The thickness of the material used as the porous surface can
be chosen depending on the application. However, it is common that
these thicknesses are about 1.0 mm or less on average, and
typically in the range of about 0.10 mm to about 1.0 mm.
[0191] Other particular contemplated porous surfaces include
grafts, particularly grafts having textured exterior portions.
Examples of textured grafts include those that have velour-textured
exteriors, with textured or smooth interiors. Grafts constructed
from woven textile products are well known in the art and have been
described in numerous documents, for example, U.S. Pat. No.
4,047,252; U.S. Pat. No. 5,178,630; U.S. Pat. No. 5,282,848; and
U.S. Pat. No. 5,800,514.
[0192] The natural biodegradable polysaccharide can be used to
provide a sealant coating to a wide variety of articles. As used
herein, "article" is used in its broadest sense and includes
objects such as devices. Such articles include, but are not limited
to vascular implants and grafts, grafts, surgical devices;
synthetic prostheses; vascular prosthesis including endoprosthesis,
stent-graft, and endovascular-stent combinations; small diameter
grafts, abdominal aortic aneurysm grafts; wound dressings and wound
management device; hemostatic barriers; mesh and hernia plugs;
patches, including uterine bleeding patches, atrial septic defect
(ASD) patches, patent foramen ovale (PFO) patches, ventricular
septal defect (VSD) patches, and other generic cardiac patches;
ASD, PFO, and VSD closures; percutaneous closure devices, mitral
valve repair devices; left atrial appendage filters; valve
annuloplasty devices, catheters; central venous access catheters,
vascular access catheters, abscess drainage catheters, drug
infusion catheters, parental feeding catheters, intravenous
catheters (e.g., treated with antithrombotic agents), stroke
therapy catheters, blood pressure and stent graft catheters;
anastomosis devices and anastomotic closures; aneurysm exclusion
devices; biosensors including glucose sensors; birth control
devices; breast implants; cardiac sensors; infection control
devices; membranes; tissue scaffolds; tissue-related materials;
shunts including cerebral spinal fluid (CSF) shunts, glaucoma drain
shunts; dental devices and dental implants; ear devices such as ear
drainage tubes, tympanostomy vent tubes; ophthalmic devices; cuffs
and cuff portions of devices including drainage tube cuffs,
implanted drug infusion tube cuffs, catheter cuff, sewing cuff;
spinal and neurological devices; nerve regeneration conduits;
neurological catheters; neuropatches; orthopedic devices such as
orthopedic joint implants, bone repair/augmentation devices,
cartilage repair devices; urological devices and urethral devices
such as urological implants, bladder devices, renal devices and
hemodialysis devices, colostomy bag attachment devices; biliary
drainage products.
[0193] In some aspects, the polymeric compositions can be utilized
in connection with an ophthalmic article. The ophthalmic article
can be configured for placement at an external or internal site of
the eye. Suitable ophthalmic articles in accordance with these
aspects can provide bioactive agent to any desired area of the eye.
In some aspects, the articles can be utilized to deliver bioactive
agent to an anterior segment of the eye (in front of the lens),
and/or a posterior segment of the eye (behind the lens). Suitable
ophthalmic devices can also be utilized to provide bioactive agent
to tissues in proximity to the eye, when desired. The biodegradable
polysaccharide compositions can be used either for the formation of
a coating on the surface of an ophthalmic article, or in the
construction of an ophthalmic article.
[0194] Suitable external articles can be configured for topical
administration of bioactive agent. Such external devices can reside
on an external surface of the eye, such as the cornea (for example,
contact lenses) or bulbar conjunctiva. In some embodiments,
suitable external devices can reside in proximity to an external
surface of the eye.
[0195] Articles configured for placement at an internal site of the
eye can reside within any desired area of the eye. In some aspects,
the ophthalmic article can be configured for placement at an
intraocular site, such as the vitreous. Illustrative intraocular
devices include, but are not limited to, those described in U.S.
Pat. Nos. 6,719,750 B2 ("Devices for Intraocular Drug Delivery,"
Varner et al.) and 5,466,233 ("Tack for Intraocular Drug Delivery
and Method for Inserting and Removing Same," Weiner et al.); U.S.
Publication Nos. 2005/0019371 A1 ("Controlled Release Bioactive
Agent Delivery Device," Anderson et al.), 2004/0133155 A1 ("Devices
for Intraocular Drug Delivery," Varner et al.), 2005/0059956 A1
("Devices for Intraocular Drug Delivery," Varner et al.), and U.S.
application Ser. Nos. 11/204,195 (filed Aug. 15, 2005, Anderson et
al.), 11/204,271 (filed Aug. 15, 2005, Anderson et al.), 11/203,981
(filed Aug. 15, 2005, Anderson et al.), 11/203,879 (filed Aug. 15,
2005, Anderson et al.), 11/203,931 (filed Aug. 15, 2005, Anderson
et al.); and related applications.
[0196] In some aspects of the invention, the biodegradable
polysaccharide coating is included on a non-linear intraocular
device. In some aspects of the invention, the biodegradable
polysaccharide coating includes a bioactive agent, such as a high
molecular weight bioactive agent useful for treating an ocular
condition.
[0197] In some aspects, the ophthalmic article can be configured
for placement, or can be formed, at a subretinal area within the
eye. Illustrative ophthalmic devices for subretinal application
include, but are not limited to, those described in U.S. Patent
Publication No. 2005/0143363 ("Method for Subretinal Administration
of Therapeutics Including Steroids; Method for Localizing
Pharmacodynamic Action at the Choroid and the Retina; and Related
Methods for Treatment and/or Prevention of Retinal Diseases," de
Juan et al.); U.S. application Ser. No. 11/175,850 ("Methods and
Devices for the Treatment of Ocular Conditions," de Juan et al.);
and related applications.
[0198] In some aspects, the invention provides a biodegradable
implant that is formed from the biodegradable polysaccharide and
that includes a bioactive agent, such as a high molecular weight
bioactive agent useful for treating an ocular condition.
[0199] In some aspects, the invention provides a method for forming
an article from the biodegradable polysaccharide, wherein the
method includes polymerizing a composition that includes the
biodegradable polysaccharide within the eye, such as in a
subretinal area or within the vitreous. For example, a low
viscosity composition including a natural biodegradable
polysaccharide and a redox pair to promote polymerization for in
situ matrix formation.
[0200] Ophthalmic articles can also be configured for placement
within any desired tissues of the eye. For example, ophthalmic
devices can be configured for placement at a subconjunctival area
of the eye, such as devices positioned extrasclerally but under the
conjunctiva, such as glaucoma drainage devices and the like.
[0201] A medical article having a biodegradable coating can also be
prepared by assembling an article having two or more "parts" (for
example, pieces of a medical article that can be put together to
form the article) wherein at least one of the parts has a
biodegradable coating. All or a portion of the part of the medical
article can have a biodegradable coating. In this regard, the
invention also contemplates parts of medical article (for example,
not the fully assembled article) that have a natural biodegradable
polysaccharide-based coating.
[0202] In some aspects of the invention the natural biodegradable
polymer is used to form the body member of a medical implant,
wherein the body member has a wet weight of about 10 g or less, or
a dry weight of about 2.5 g or less.
[0203] The device can also have a base coating of material. The
base coating can serve one or more functions, for example, it can
provide an improved surface for the natural biodegradable
polysaccharide or composition that includes the natural
biodegradable polysaccharide. The base coating can include a
polymeric material, such as a natural or synthetic polymer.
Examples of suitable compounds that can be used to pretreat a
surface to provide a base coat include Parylene and organosilane
compounds. Suitable base coatings can include, for example,
methacrylate, acrylate, alkylacrylate, acrylamide,
vinylpyrrolidinone, vinylacetamide, and vinyl formamide based
polymers and copolymers. These polymers can also include latent
reactive groups such as photoreactive groups.
[0204] Base coatings can be useful in various coating processes.
For example, in some aspects, biodegradable microparticles can be
first disposed on a base coat and then the natural biodegradable
polysaccharide having coupling groups can be disposed on the
microparticles. The surface can then be treated to form a coating
wherein the microparticles are predominantly located between the
base layer and the layer formed from the natural biodegradable
polysaccharide having coupling groups. If desired, an initiator can
be included in a base coating and the natural biodegradable
polysaccharide polymer or composition that includes the natural
biodegradable polysaccharide polymer can be disposed on the base
coating. The base coating can serve one or more functions, for
example, it can provide an improved surface for the natural
biodegradable polysaccharide or composition that includes the
natural biodegradable polysaccharide.
[0205] In many aspects of the invention, the natural biodegradable
polysaccharide coating or the biodegradable article includes one or
more bioactive agents. The bioactive agent can be dispersed within
the natural biodegradable polysaccharide coating or biodegradable
article itself. Alternatively, the bioactive agent can be present
in microparticles that are associated with the natural
biodegradable polysaccharide coating. The bioactive agent can be
delivered from the coated surface upon degradation of the natural
biodegradable polysaccharide and/or biodegradable
microparticles.
[0206] The term "bioactive agent" refers to a peptide, protein,
carbohydrate, nucleic acid, lipid, polysaccharide, synthetic
inorganic or organic molecule, viral particle, cell, or
combinations thereof, that causes a biological effect when
administered in vivo to an animal, including but not limited to
birds and mammals, including humans. Nonlimiting examples are
antigens, enzymes, hormones, receptors, peptides, and gene therapy
agents. Examples of suitable gene therapy agents include (a)
therapeutic nucleic acids, including antisense DNA, antisense RNA,
and interference RNA, and (b) nucleic acids encoding therapeutic
gene products, including plasmid DNA and viral fragments, along
with associated promoters and excipients. Examples of other
molecules that can be incorporated include nucleosides,
nucleotides, vitamins, minerals, and steroids.
[0207] Although not limited to such, the coatings of the invention
are particularly useful for delivering bioactive agents that are
large hydrophilic molecules, such as polypeptides (including
proteins and peptides), nucleic acids (including DNA and RNA),
polysaccharides (including heparin), as well as particles, such as
viral particles, and cells. In one aspect, the bioactive agent has
a molecular weight of about 10,000 or greater.
[0208] Classes of bioactive agents which can be incorporated into
biodegradable coatings (both the natural biodegradable matrix
and/or the biodegradable microparticles) of this invention include,
but are not limited to: ACE inhibitors, actin inhibitors,
analgesics, anesthetics, anti-hypertensives, anti polymerases,
antisecretory agents, anti-AIDS substances, antibiotics,
anti-cancer substances, anti-cholinergics, anti-coagulants,
anti-convulsants, anti-depressants, anti-emetics, antifungals,
anti-glaucoma solutes, antihistamines, antihypertensive agents,
anti-inflammatory agents (such as NSAIDs), anti metabolites,
antimitotics, antioxidizing agents, anti-parasite and/or
anti-Parkinson substances, antiproliferatives (including
antiangiogenesis agents), anti-protozoal solutes, anti-psychotic
substances, anti-pyretics, antiseptics, anti-spasmodics, antiviral
agents, calcium channel blockers, cell response modifiers,
chelators, chemotherapeutic agents, dopamine agonists,
extracellular matrix components, fibrinolytic agents, free radical
scavengers, growth hormone antagonists, hypnotics,
immunosuppressive agents, immunotoxins, inhibitors of surface
glycoprotein receptors, microtubule inhibitors, miotics, muscle
contractants, muscle relaxants, neurotoxins, neurotransmitters,
opioids, photodynamic therapy agents, prostaglandins, remodeling
inhibitors, statins, steroids, thrombolytic agents, tranquilizers,
vasodilators, and vasospasm inhibitors.
[0209] Antibiotics are art recognized and are substances which
inhibit the growth of or kill microorganisms. Examples of
antibiotics include penicillin, tetracycline, chloramphenicol,
minocycline, doxycycline, vancomycin, bacitracin, kanamycin,
neomycin, gentamycin, erythromycin, cephalosporins, geldanamycin,
and analogs thereof. Examples of cephalosporins include
cephalothin, cephapirin, cefazolin, cephalexin, cephradine,
cefadroxil, cefamandole, cefoxitin, cefaclor, cefuroxime,
cefonicid, ceforanide, cefotaxime, moxalactam, ceftizoxime,
ceftriaxone, and cefoperazone.
[0210] Antiseptics are recognized as substances that prevent or
arrest the growth or action of microorganisms, generally in a
nonspecific fashion, e.g., by inhibiting their activity or
destroying them. Examples of antiseptics include silver
sulfadiazine, chlorhexidine, glutaraldehyde, peracetic acid, sodium
hypochlorite, phenols, phenolic compounds, iodophor compounds,
quaternary ammonium compounds, and chlorine compounds.
[0211] Anti-viral agents are substances capable of destroying or
suppressing the replication of viruses. Examples of anti-viral
agents include .alpha.-methyl-P-adamantane methylamine,
hydroxy-ethoxymethylguanine, adamantanamine,
5-iodo-2'-deoxyuridine, trifluorothymidine, interferon, and adenine
arabinoside.
[0212] Enzyme inhibitors are substances that inhibit an enzymatic
reaction. Examples of enzyme inhibitors include edrophonium
chloride, N-methylphysostigmine, neostigmine bromide, physostigmine
sulfate, tacrine HCl, tacrine, 1-hydroxymaleate, iodotubercidin,
p-bromotetramisole,
10-(.alpha.-diethylaminopropionyl)-phenothiazine hydrochloride,
calmidazolium chloride, hemicholinium-3,3,5-dinitrocatechol,
diacylglycerol kinase inhibitor I, diacylglycerol kinase inhibitor
II, 3-phenylpropargylamine, N-monomethyl-L-arginine acetate,
carbidopa, 3-hydroxybenzylhydrazine HCl, hydralazine HCl,
clorgyline HCl, deprenyl HCl, L(-), deprenyl HCl, D(+),
hydroxylamine HCl, iproniazid phosphate,
6-MeO-tetrahydro-9H-pyrido-indole, nialamide, pargyline HCl,
quinacrine HCl, semicarbazide HCl, tranylcypromine HCl,
N,N-diethylaminoethyl-2,2-diphenylvalerate hydrochloride,
3-isobutyl-1-methylxanthine, papaverine HCl, indomethacin,
2-cyclooctyl-2-hydroxyethylamine hydrochloride,
2,3-dichloro-.alpha.-methylbenzylamine (DCMB),
8,9-dichloro-2,3,4,5-tetrahydro-1H-2-benzazepine hydrochloride,
p-aminoglutethimide, p-aminoglutethimide tartrate, R(+),
p-aminoglutethimide tartrate, S(-), 3-iodotyrosine,
alpha-methyltyrosine, L(-) alpha-methyltyrosine, D L(-),
cetazolamide, dichlorphenamide,
6-hydroxy-2-benzothiazolesulfonamide, and allopurinol.
[0213] Anti-pyretics are substances capable of relieving or
reducing fever. Anti-inflammatory agents are substances capable of
counteracting or suppressing inflammation. Examples of such agents
include aspirin (salicylic acid), indomethacin, sodium indomethacin
trihydrate, salicylamide, naproxen, colchicine, fenoprofen,
sulindac, diflunisal, diclofenac, indoprofen and sodium
salicylamide. Local anesthetics are substances that have an
anesthetic effect in a localized region. Examples of such
anesthetics include procaine, lidocaine, tetracaine and
dibucaine.
[0214] Cell response modifiers are chemotactic factors such as
platelet-derived growth factor (pDGF). Other chemotactic factors
include neutrophil-activating protein, monocyte chemoattractant
protein, macrophage-inflammatory protein, SIS (small inducible
secreted) proteins, platelet factor, platelet basic protein,
melanoma growth stimulating activity, epidermal growth factor,
transforming growth factor (alpha), fibroblast growth factor,
platelet-derived endothelial cell growth factor, insulin-like
growth factor, nerve growth factor, and bone
growth/cartilage-inducing factor (alpha and beta). Other cell
response modifiers are the interleukins, interleukin inhibitors or
interleukin receptors, including interleukin 1 through interleukin
10; interferons, including alpha, beta and gamma; hematopoietic
factors, including erythropoietin, granulocyte colony stimulating
factor, macrophage colony stimulating factor and
granulocyte-macrophage colony stimulating factor; tumor necrosis
factors, including alpha and beta; transforming growth factors
(beta), including beta-1, beta-2, beta-3, inhibin, activin, and DNA
that encodes for the production of any of these proteins.
[0215] Examples of statins include lovastatin, pravastatin,
simvastatin, fluvastatin, atorvastatin, cerivastatin, rousvastatin,
and superstatin.
[0216] Imaging agents are agents capable of imaging a desired site,
e.g., tumor, in vivo, can also be included in the coating
composition. Examples of imaging agents include substances having a
label which is detectable in vivo, e.g., antibodies attached to
fluorescent labels. The term antibody includes whole antibodies or
fragments thereof.
[0217] Exemplary ligands or receptors include antibodies, antigens,
avidin, streptavidin, biotin, heparin, type IV collagen, protein A,
and protein G.
[0218] Exemplary antibiotics include antibiotic peptides.
[0219] In some aspects the bioactive agent can be selected to
improve the compatibility (for example, with blood and/or
surrounding tissues) of medical device surfaces. These agents,
referred to herein as "biocompatible agents," when associated with
the medical device surface, can serve to shield the blood from the
underlying medical device material. Suitable biocompatible agents
preferably reduce the likelihood for blood components to adhere to
the medical device, thus reducing the formation of thrombus or
emboli (blood clots that release and travel downstream).
[0220] The bioactive agent can improve the biocompatibility of the
medical article having a coating that includes the natural
biodegradable polymer and the biodegradable microparticle. The
bioactive agent can provide antirestenotic effects, such as
antiproliferative, anti-platelet, and/or antithrombotic effects. In
some embodiments, the bioactive agent can include anti-inflammatory
agents, immunosuppressive agents, cell attachment factors,
receptors, ligands, growth factors, antibiotics, enzymes, nucleic
acids, and the like. Compounds having antiproliferative effects
include, for example, actinomycin D, angiopeptin, c-myc antisense,
paclitaxel, taxane, and the like.
[0221] Representative examples of bioactive agents having
antithrombotic effects include heparin, heparin derivatives, sodium
heparin, low molecular weight heparin, hirudin, lysine,
prostaglandins, argatroban, forskolin, vapiprost, prostacyclin and
prostacyclin analogs, D-ph-pr-arg-chloromethylketone (synthetic
antithrombin), dipyridamole, glycoprotein IIb/IIIa platelet
membrane receptor antibody, coprotein IIb/IIIa platelet membrane
receptor antibody, recombinant hirudin, thrombin inhibitor (such as
commercially available from Biogen), chondroitin sulfate, modified
dextran, albumin, streptokinase, tissue plasminogen activator
(TPA), urokinase, nitric oxide inhibitors, and the like.
[0222] The bioactive agent can also be an inhibitor of the
GPIIb-IIIa platelet receptor complex, which mediates platelet
aggregation. GPIIb/IIIa inhibitors can include monoclonal antibody
Fab fragment c7E3, also know as abciximab (ReoPro.TM.), and
synthetic peptides or peptidomimetics such as eptifibatide
(Integrilin.TM.) or tirofiban (Agrastat.TM.).
[0223] The bioactive agent can be an immunosuppressive agent, for
example, cyclosporine, CD-34 antibody, everolimus, mycophenolic
acid, sirolimus, tacrolimus, and the like.
[0224] Other exemplary therapeutic antibodies include trastuzumab
(Herceptin.TM.), a humanized anti-HER2 monoclonal antibody (moAb);
alemtuzumab (Campath.TM.), a humanized anti-CD52 moAb; gemtuzumab
(Mylotarg.TM.), a humanized anti-CD33 moAb; rituximab
(Rituxan.TM.), a chimeric anti-CD20 moAb; ibritumomab
(Zevalin.TM.), a murine moAb conjugated to a beta-emitting
radioisotope; tositumomab (Bexxar.TM.), a murine anti-CD20 moAb;
edrecolomab (Panorex.TM.), a murine anti-epithelial cell adhesion
molecule moAb; cetuximab (Erbitux.TM.), a chimeric anti-EGFR moAb;
and bevacizumab (Avastin.TM.), a humanized anti-VEGF moAb.
[0225] Additionally, the bioactive agent can be a surface adhesion
molecule or cell-cell adhesion molecule. Exemplary cell adhesion
molecules or attachment proteins (such as extracellular matrix
proteins including fibronectin, laminin, collagen, elastin,
vitronectin, tenascin, fibrinogen, thrombospondin, osteopontin, von
Willibrand Factor, bone sialoprotein (and active domains thereof),
or a hydrophilic polymer such as hyaluronic acid, chitosan or
methyl cellulose, and other proteins, carbohydrates, and fatty
acids. Exemplary cell-cell adhesion molecules include N-cadherin
and P-cadherin and active domains thereof.
[0226] Exemplary growth factors include fibroblastic growth
factors, epidermal growth factor, platelet-derived growth factors,
transforming growth factors, vascular endothelial growth factor,
bone morphogenic proteins and other bone growth factors, and neural
growth factors.
[0227] The bioactive agent can be also be selected from
mono-2-(carboxymethyl) hexadecanamidopoly (ethylene glycol).sub.200
mono-4-benzoylbenzyl ether, mono-3-carboxyheptadecanamidopoly
(ethylene glycol).sub.200 mono-4-benzoylbenzyl ether,
mono-2-(carboxymethyl) hexadecanamidotetra (ethylene glycol)
mono-4-benzoylbenzyl ether, mono-3-carboxyheptadecanamidotetra
(ethylene glycol) mono-4-benzoylbenzyl ether,
N-[2-(4-benzoylbenzyloxy) ethyl]-2-(carboxymethyl) hexadecanamide,
N-[2-(4-benzoylbenzyloxy)ethyl]-3-carboxyheptadecanamide,
N-[12-(benzoylbenzyloxy) dodecyl]-2-(carboxymethyl) hexadecanamide,
N-[12-(benzoylbenzyloxy) dodecyl]-3-carboxy-heptadecanamide,
N-[3-(4-benzoylbenzamido) propyl]-2-(carboxymethyl) hexadecanamide,
N-[3-(4-benzoylbenzamido) propyl]-3-carboxyheptadecanamide,
N-(3-benzoylphenyl)-2-(carboxymethyl) hexadecanamide,
N-(3-benzoylphenyl)-3-carboxyheptadecanamide,
N-(4-benzoylphenyl)-2-(carboxymethyl) hexadecanamide, poly(ethylene
glycol).sub.200 mono-15-carboxypentadecyl mono-4-benzoylbenzyl
ether, and mono-15-carboxypentadecanamidopoly (ethylene
glycol).sub.200 mono-4-benzoylbenzyl ether.
[0228] Additional examples of contemplated bioactive agents and/or
bioactive agent include analogues of rapamycin ("rapalogs"),
ABT-578 from Abbott, dexamethasone, betamethasone, vinblastine,
vincristine, vinorelbine, poside, teniposide, daunorubicin,
doxorubicin, idarubicin, anthracyclines, mitoxantrone, bleomycins,
plicamycin (mithramycin), mitomycin, mechlorethamine,
cyclophosphamide and its analogs, melphalan, chlorambucil,
ethylenimines and methylmelamines, alkyl sulfonates-busulfan,
nitrosoureas, carmustine (BCNU) and analogs, streptozocin,
trazenes-dacarbazinine, methotrexate, fluorouracil, floxuridine,
cytarabine, mercaptopurine, thioguanine, pentostatin,
2-chlorodeoxyadenosine, cisplatin, carboplatin, procarbazine,
hydroxyurea, mitotane, estrogen, ticlopidine, clopidogrel,
abciximab, breveldin, cortisol, cortisone, fludrocortisone,
prednisone, prednisolone, 6U-methylprednisolone, triamcinolone,
acetaminophen, etodalac, tolmetin, ketorolac, ibuprofen and
derivatives, mefenamic acid, meclofenamic acid, piroxicam,
tenoxicam, phenylbutazone, oxyphenthatrazone, nabumetone,
auranofin, aurothioglucose, gold sodium thiomalate, azathioprine,
mycophenolate mofetil; angiotensin receptor blockers; nitric oxide
donors; and mTOR inhibitors.
[0229] Viral particles and viruses include those that may be
therapeutically useful, such as those used for gene therapy, and
also attenuated viral particles and viruses which can promote an
immune response and generation of immunity. Useful viral particles
include both natural and synthetic types. Viral particles include,
but are not limited to, adenoviruses, baculoviruses, parvoviruses,
herpesviruses, poxviruses, adeno-associated viruses, vaccinia
viruses, and retroviruses.
[0230] Other bioactive agents that can be used for altering gene
function include plasmids, phages, cosmids, episomes, and
integratable DNA fragments, antisense oligonucleotides, antisense
DNA and RNA, modified DNA and RNA, iRNA, ribozymes, siRNA, and
shRNA.
[0231] Other bioactive agents include cells such as platelets, stem
cells, T lymphocytes, B lymphocytes, acidophils, adipocytes,
astrocytes, basophils, hepatocytes, neurons, cardiac muscle cells,
chondrocytes, epithelial cells, dendrites, endrocrine cells,
endothelial cells, eosinophils, erythrocytes, fibroblasts,
follicular cells, ganglion cells, hepatocytes, endothelial cells,
Leydig cells, parenchymal cells, lymphocytes, lysozyme-secreting
cells, macrophages, mast cells, megakaryocytes, melanocytes,
monocytes, myoid cells, neck nerve cells, neutrophils,
oligodendrocytes, oocytes, osteoblasts, osteochondroclasts,
osteoclasts, osteocytes, plasma cells, spermatocytes,
reticulocytes, Schwann cells, Sertoli cells, skeletal muscle cells,
and smooth muscle cells. Bioactive agents can also include
genetically modified, recombinant, hybrid, mutated cells, and cells
with other alterations.
[0232] Additives such as inorganic salts, BSA (bovine serum
albumin), and inert organic compounds can be used to alter the
profile of bioactive agent release, as known to those skilled in
the art.
[0233] The concentration of the bioactive agent or agents dissolved
or suspended in the coating mixture can range from about 0.01 to
about 90 percent, by weight, based on the weight of the final
coated composition.
[0234] The particular bioactive agent, or combination of bioactive
agents, can be selected depending upon one or more of the following
factors: the application of the controlled delivery device, the
medical condition to be treated, the anticipated duration of
treatment, characteristics of the implantation site, the number and
type of bioactive agents to be utilized, and the like.
[0235] Any of the polymer compositions described herein can be
provided to the surface of the medical article and can include any
number of desired bioactive agents, depending upon the final
application of the medical device.
[0236] A comprehensive listing of bioactive agents can be found in
The Merck Index, Thirteenth Edition, Merck & Co. (2001).
Bioactive agents are commercially available from Sigma Aldrich Fine
Chemicals, Milwaukee, Wis.
[0237] In some aspects of the invention, the bioactive agent can be
used to promote thrombosis in association with the natural
biodegradable polysaccharide-based coating, which can be of
particular use when a coating having a sealant function is desired.
A sealant coating including a thrombogenic agent can promote the
in-growth of tissue upon degradation of the sealant coating
material. The degree of thrombosis can be controlled by various
factors, including, for example, the presence of one or more
thrombosis-promoting bioactive agents on or within the coating.
Suitable thrombotic agents are described herein.
[0238] In some aspects the thrombotic agent can be selected to have
an affect on the blood and/or surrounding tissues that are in
contact with the article surface. In some cases the thrombotic
agent is chosen for the ability to affect the ability of blood
components to adhere to the medical article. The thrombotic agent
can, in some cases, be chosen to promote thrombus formation at the
surface of the coated article. Therefore, in some embodiments, the
sealant coating can include a thrombotic agent, such as thrombin,
collagen (for example, (synthetic) recombinant human collagen
(FibroGen, South San Francisco, Calif.)), ADP, or convulxin to
promote thrombosis at the coated surface of the article.
[0239] Other prothrombotic or procoagulant factors include platelet
factors 1-4, platelet activating factor (acetyl glyceryl ether
phosphoryl choline); P-selectin and von Willebrand Factor (vWF);
tissue factor; plasminogen activator initiator-1; thromboxane;
procoagulant thrombin-like enzymes including cerastotin and
afaacytin; phospholipase A.sub.2; Ca.sup.2+-dependent lectins
(C-type lectin); factors that bind glycoprotein receptors and
induce aggregation including aggretin, rhodocytin,
aggregoserpentin, triwaglerin, and equinatoxin; glycoprotein Ib
agonists including mamushigin and alboaggregin; vWF interacting
factors including botrocetin, bitiscetin, cerastotin, and
ecarin.
[0240] Other factors, including protein factors, that are involved
in the clotting cascade include coagulation factors I-XIII (for
example, fibrinogen, prothrombin, tissue thromboplastin, calcium,
proaccelerin (accelerator globulin), proconvertin (serum
prothrombin conversion accelerator), antihemophilic factor, plasma
thromboplastin component, Stuart factor (autoprothrombin C), plasma
thromboplastin antecedent (PTA), Hageman factor, and
fibrin-stabilizing factor (FSF, fibrinase,
protransglutaminase)).
[0241] Some surface adhesion molecule or cell-cell adhesion
molecules may also function to promote coagulation or thrombosis.
Exemplary cell adhesion molecules or attachment proteins (such as
extracellular matrix proteins) include fibronectin, laminin,
collagen, elastin, vitronectin, tenascin, fibrinogen,
thrombospondin, osteopontin, von Willebrand Factor, bone
sialoprotein (and active domains thereof), or a hydrophilic polymer
such as hyaluronic acid, chitosan or methyl cellulose, and other
proteins, carbohydrates, and fatty acids. Exemplary cell-cell
adhesion molecules include N-cadherin and P-cadherin and active
domains thereof.
[0242] The particular thrombotic agent, or a combination of
thrombotic agents with other bioactive agents, can be selected
depending upon one or more of the following factors: the
application of the medical article, the medical condition to be
treated, the anticipated duration of treatment, characteristics of
the implantation site, the number and type of
thrombogenic/bioactive agents to be utilized, the chemical
composition of the sealant coating (such as amylose, selected
additives, and the like), the extent of coupling in the formed
sealant coating, and the like.
[0243] Any of the sealant compositions described herein can be
provided to the surface of the medical article. In some embodiments
the sealant coating can include any number of desired
thrombogenic/bioactive agents, depending upon the final application
of the medical article. The coating of sealant material (with or
without thrombogenic/bioactive agents) can be applied to the
medical article using standard techniques to cover the entire
surface of the article, or a portion of the article surface.
Further, the sealant composition material can be provided as a
single coated layer (with or without thrombogenic/bioactive
agents), or as multiple coated layers (with or without
thrombogenic/bioactive agents). When multiple coated layers are
provided on the surface, the materials of each coated layer can be
chosen to provide a desired effect.
[0244] In some aspects of the invention, a microparticle is used to
deliver the bioactive agent from the natural biodegradable
polysaccharide-based coating. The microparticles of the invention
can comprise any three-dimensional structure that can be
immobilized on a substrate in association with the matrix formed by
the amylose polymer. The term "microparticle" is intended to
reflect that the three-dimensional structure is very small but not
limited to a particular size range, or not limited to a structure
that has a particular shape. According to the invention,
microparticles typically have a size in the range of 5 nm to 100
.mu.m in diameter. Generally microparticles are spherical or
somewhat spherical in shape, but can have other shapes as well. In
preferred embodiments of the invention, the biodegradable
microparticles have a size in the range of 100 nm to 20 .mu.m in
diameter, and even more preferable in the range of 400 nm to 20
.mu.m in diameter.
[0245] The microparticle being "biodegradable" refers to the
presence of one or more biodegradable materials in the
microparticle. The biodegradable microparticles include at least a
biodegradable material (such as a biodegradable polymer) and a
bioactive agent. The biodegradable microparticles can gradually
decompose and release bioactive agent upon exposure to an aqueous
environment, such as body fluids.
[0246] The biodegradable microparticle can also include one or more
biodegradable polymers. Examples of biodegradable polymers that can
be included in the biodegradable microparticle include, for
example, polylactic acid, poly(lactide-co-glycolide),
polycaprolactone, polyphosphazine, polymethylidenemalonate,
polyorthoesters, polyhydroxybutyrate, polyalkeneanhydrides,
polypeptides, polyanhydrides, and polyesters, and the like.
[0247] Biodegradable polyetherester copolymers can be used.
Generally speaking, the polyetherester copolymers are amphiphilic
block copolymers that include hydrophilic (for example, a
polyalkylene glycol, such as polyethylene glycol) and hydrophobic
blocks (for example, polyethylene terephthalate). Examples of block
copolymers include poly(ethylene glycol)-based and poly(butylene
terephthalate)-based blocks (PEG/PBT polymer). Examples of these
types of multiblock copolymers are described in, for example, U.S.
Pat. No. 5,980,948. PEG/PBT polymers are commercially available
from Octoplus BV, under the trade designation PolyActive.TM..
[0248] Biodegradable copolymers having a biodegradable, segmented
molecular architecture that includes at least two different ester
linkages can also be used. The biodegradable polymers can be block
copolymers (of the AB or ABA type) or segmented (also known as
multiblock or random-block) copolymers of the (AB).sub.n type.
These copolymers are formed in a two (or more) stage ring opening
copolymerization using two (or more) cyclic ester monomers that
form linkages in the copolymer with greatly different
susceptibilities to transesterification. Examples of these polymers
are described in, for example, in U.S. Pat. No. 5,252,701 (Jarrett
et al., "Segmented Absorbable Copolymer").
[0249] Other suitable biodegradable polymer materials include
biodegradable terephthalate copolymers that include a
phosphorus-containing linkage. Polymers having phosphoester
linkages, called poly(phosphates), poly(phosphonates) and
poly(phosphites), are known. See, for example, Penczek et al.,
Handbook of Polymer Synthesis, Chapter 17: "Phosphorus-Containing
Polymers," 1077-1132 (Hans R. Kricheldorf ed., 1992), as well as
U.S. Pat. Nos. 6,153,212, 6,485,737, 6,322,797, 6,600,010,
6,419,709. Biodegradable terephthalate polyesters can also be used
that include a phosphoester linkage that is a phosphite. Suitable
terephthalate polyester-polyphosphite copolymers are described, for
example, in U.S. Pat. No. 6,419,709 (Mao et al., "Biodegradable
Terephthalate Polyester-Poly(Phosphite) Compositions, Articles, and
Methods of Using the Same). Biodegradable terephthalate polyester
can also be used that include a phosphoester linkage that is a
phosphonate. Suitable terephthalate polyester-poly(phosphonate)
copolymers are described, for example, in U.S. Pat. Nos. 6,485,737
and 6,153,212 (Mao et al., "Biodegradable Terephthalate
Polyester-Poly(Phosphonate) Compositions, Articles and Methods of
Using the Same). Biodegradable terephthalate polyesters can be used
that include a phosphoester linkage that is a phosphate. Suitable
terephthalate polyester-poly(phosphate) copolymers are described,
for example, in U.S. Pat. Nos. 6,322,797 and 6,600,010 (Mao et al.,
"Biodegradable Terephthalate Polyester-Poly(Phosphate) Polymers,
Compositions, Articles, and Methods for Making and Using the
Same).
[0250] Biodegradable polyhydric alcohol esters can also be used
(See U.S. Pat. No. 6,592,895). This patent describes biodegradable
star-shaped polymers that are made by esterifying polyhydric
alcohols to provide acyl moieties originating from aliphatic
homopolymer or copolymer polyesters. The biodegradable polymer can
be a three-dimensional crosslinked polymer network containing
hydrophobic and hydrophilic components which forms a hydrogel with
a crosslinked polymer structure, such as that described in U.S.
Pat. No. 6,583,219. The hydrophobic component is a hydrophobic
macromer with unsaturated group terminated ends, and the
hydrophilic polymer is a polysaccharide containing hydroxy groups
that are reacted with unsaturated group introducing compounds. The
components are convertible into a one-phase crosslinked polymer
network structure by free radical polymerization. In yet further
embodiments, the biodegradable polymer can comprise a polymer based
upon .alpha.-amino acids (such as elastomeric copolyester amides or
copolyester urethanes, as described in U.S. Pat. No.
6,503,538).
[0251] The biodegradable microparticle can include one or more
biodegradable polymers obtained from natural sources. In some
preferred aspects the biodegradable polymer is selected from
hyaluronic acid, dextran, starch, amylose, amylopectin, cellulose,
xanthan, pullulan, chitosan, pectin, inulin, alginates, and
heparin. One, or combinations of more than one of these
biodegradable polymers, can be used. A particular biodegradable
polymer can also be selected based on the type of bioactive agent
that is present in the microparticle. Therefore, in some aspects of
the invention, the biodegradable coating can include a natural
biodegradable polysaccharide matrix and a natural biodegradable
polysaccharide-containing microparticle.
[0252] Therefore, in some embodiments, the microparticles include a
natural biodegradable polysaccharide such as amylose or
maltodextrin. In some embodiments the natural biodegradable
polysaccharide can be the primary biodegradable component in the
microparticle. In some embodiments, both the coating matrix and the
microparticle include amylose and/or maltodextrin as
components.
[0253] Dextran-based microparticles can be particularly useful for
the incorporation of bioactive agents such as proteins, peptides,
and nucleic acids. Examples of the preparation of dextran-based
microparticles are described in U.S. Pat. No. 6,303,148.
[0254] The preparation of amylose and other starch-based
microparticles have been described in various references,
including, for example, U.S. Pat. No. 4,713,249; U.S. Pat. No.
6,692,770; and U.S. Pat. No. 6,703,048. Biodegradable polymers and
their synthesis have been also been described in various references
including Mayer, J. M., and Kaplan, D. L. (1994) Trends in Polymer
Science 2: pages 227-235; and Jagur-Grodzinski, J., (1999) Reactive
and Functional Polymers: Biomedical Application of Functional
Polymers, Vol. 39, pages 99-138.
[0255] In some aspects of the invention, the biodegradable
microparticle contains a biologically active agent (a "bioactive
agent"), such as a pharmaceutical or a prodrug. Microparticles can
be prepared incorporating various bioactive agents by established
techniques, for example, by solvent evaporation (see, for example,
Wichert, B. and Rohdewald, P. J. Microencapsul. (1993) 10:195). The
bioactive agent can be released from the biodegradable
microparticle (the microparticle being present in the natural
biodegradable polysaccharide coating) upon degradation of the
biodegradable microparticle in vivo. Microparticles having
bioactive agent can be formulated to release a desired amount of
the agent over a predetermined period of time. It is understood
that factors affecting the release of the bioactive agent and the
amount released can be altered by the size of the microparticle,
the amount of bioactive agent incorporated into the microparticle,
the type of degradable material used in fabricating the
microparticle, the amount of biodegradable microparticles
immobilized per unit area on the substrate, etc.
[0256] The microparticles can also be treated with a porogen, such
as salt, sucrose, PEG, or an alcohol, to create pores of a desired
size for incorporation of the bioactive agent.
[0257] The quantity of bioactive agents provided in the
biodegradable microparticle can be adjusted by the user to achieve
the desired effect. For example, a particular amount of
anti-coagulant drug can be incorporated into the microparticle to
provide a certain level of anti-coagulant activity from the
biodegradable coating. Biologically active compounds can be
provided by the microparticles in a range suitable for the
application. In another example, protein molecules can be provided
by biodegradable microparticles. For example, the amount of protein
molecules present can be in the range of 1-250,000 molecules per 1
.mu.m diameter microparticle.
[0258] Generally, the concentration of the bioactive agent present
in the biodegradable microparticles can be chosen based on any one
or a combination of a number of factors, including, but not limited
to, the release rate from the coating, the type of bioactive
agent(s) in the coating, the desired local or systemic
concentration of the bioactive agent following release, and the
half life of the bioactive agent. In some cases the concentration
of bioactive agent in the microparticle can be about 0.001% or
greater, or in the range of about 0.001% to about 50 percent, or
greater, by weight, based on the weight of the microparticle.
[0259] The particular bioactive agent to be included in the
biodegradable microparticle, or combination of bioactive agents in
microparticles, can be selected depending upon factors such as the
application of the coated device, the medical condition to be
treated, the anticipated duration of treatment, characteristics of
the implantation site, the number and type of bioactive agents to
be utilized, the chemical composition of the microparticle, size of
the microparticle, crosslinking, and the like.
[0260] In one embodiment, the invention advantageously allows for
preparation of surfaces having two, or more than two, different
bioactive agents, wherein the bioactive agents are mutually
incompatible in a particular environment, for example, as
hydrophobic and hydrophilic drugs are incompatible in either a
polar or non-polar solvent. Different bioactive agents may also
demonstrate incompatibility based on protic/aprotic solvents or
ionic/non-ionic solvents. For example, the invention allows for the
preparation of one set of biodegradable microparticles containing a
hydrophobic drug and the preparation of another set of
biodegradable microparticles containing a hydrophilic drug; the
mixing of the two different sets of microparticles into a polymeric
material used to form the matrix; and the disposing of the mixture
on the surface of a substrate. Both hydrophobic and hydrophilic
drugs can be released from the surface of the coated substrate at
the same time as the biodegradable microparticles degrade, or the
composition of the biodegradable microparticles or the natural
biodegradable polysaccharide matrix can be altered so that one
bioactive agent is released at a different rate or time than the
other one.
[0261] Biodegradable microparticles can be prepared having
compositions that are suitable for either hydrophobic or
hydrophilic drugs. For example, polymers such as polylactide or
polycaprolactone can be useful for preparing biodegradable
microparticles that include hydrophobic drugs; whereas polymers
such as amylose or glycolide can be useful for preparing
microparticles that include hydrophilic drugs.
[0262] Traditional coating procedures directed at disposing at
least two different types of bioactive agents have often required
that the bioactive agents be put down separately. Traditional
approaches may include the steps of solubilizing a hydrophobic drug
in a non-polar solvent, coating the surface of the substrate with
the non-polar mixture, drying the non-polar mixture, solubilizing
the hydrophilic drug in a polar solvent, coating the layer of the
dried non-polar mixture with the polar mixture, and then drying the
polar mixture. This type of traditional coating process can be
inefficient and can also result in undesirable surface properties
(e.g., the layering of the drugs will cause one drug to be released
before the other one is released). According to this aspect of the
invention, the method of preparing surfaces having two, or more
than two, different bioactive agents, in particular when the two
different bioactive agents are released from the surface of the
substrate, is a significant improvement over traditional methods of
coating substrates and delivering bioactive agents from the surface
of the substrates.
[0263] Components of the biodegradable coating can be applied to
the medical device using standard techniques to cover the entire
surface of the device, or a portion of the device surface. As
indicated, the components can be applied to the medical device
independently or together, for example, in a composition. The
coating formed on the device can be a single layer coating, or a
multiple layer coating.
[0264] Various factors can influence the delivery of bioactive
agents from the coating. These include the concentration of the
natural biodegradable polysaccharide and the extent of natural
biodegradable polysaccharide coupling in the coating, the amount
and location of biodegradable microparticles associated with the
coating, the concentration of bioactive agent in the
microparticles, and the presence of other coated layers, if
included in the overall coating and the like. For example, the rate
of delivery of the drug can be decreased by increasing the
concentration of polymeric material or the relative amount of
coupling or crosslinking of the polymeric material in the polymeric
matrix or in the microparticle. Based on the description provided
herein and the general knowledge in this technical area, one can
alter properties of the coating to provide a desired release rate
for one or more particular bioactive agents from the coating.
[0265] Portions of the coating can be prepared to degrade at the
same or different rates. For example, the biodegradable
microparticles can be prepared or obtained to have a faster rate of
degradation than the natural biodegradable polysaccharide matrix.
In this case, the bioactive agent can be released into the natural
biodegradable polysaccharide matrix and/or diffuse out of the
natural biodegradable polysaccharide matrix.
[0266] A natural biodegradable polysaccharide-based coating can be
prepared by any one of a variety of methods. A "coating" as used
herein can include one or more "coated layers", each coated layer
including one or more coating materials. In many cases, the coating
consists of a single layer of material that includes the natural
biodegradable polysaccharide, such as amylose or maltodextrin. In
other cases, the coating includes more than one coated layer, at
least one of the coated layers including the natural biodegradable
polysaccharide. If more than one layer is present in the coating,
the layers can be composed of the same or different materials. If
multiple polymeric layers are provided on the surface, each
individual layer of polymer can be chosen to provide a desired
effect. Additionally, multiple layers of various bioactive agents
can be deposited onto the medical device surface so that a
particular bioactive agent can be presented to or released from the
medical device at one time, one or more bioactive agents in each
layer, which can be separated by polymeric material.
[0267] If more than one coated layer is applied to a surface, it is
typically applied successively. For example, a natural
biodegradable polysaccharide coated layer can be formed by, for
example, dipping, spraying, bushing, or swabbing the coating
material on the article to form a layer, and then drying the coated
layer. The process can be repeated to provide a coating having
multiple coated layers, wherein at least one layer includes the
natural biodegradable polysaccharide.
[0268] Thus, in some embodiments wherein multiple coated layers are
prepared, each coated layer is composed of the same materials.
Alternatively, one or more of the coated layers is composed of
materials that are different from one or more of the other layers.
Additionally, multiple layers of various bioactive agents can be
deposited onto the medical article surface so that a particular
bioactive agent can be presented to or released from the medical
article at one time, one or more bioactive agents in each layer,
which can be separated by polymeric material.
[0269] The invention also provides the advantage of maintaining
excellent control over the formation of a coating on the surface of
an article. To exemplify this aspect of the invention, an initiator
is disposed on a surface of a medical article along with the
natural biodegradable polysaccharide having pendent coupling
groups. A bioactive agent can be disposed if desired. The initiator
can be disposed in a mixture with the natural biodegradable
polysaccharide together, or the initiator can be disposed
independently. These compounds are generally disposed in a fluid
state (for example, suspended or dissolved in an aqueous liquid)
and can be disposed on an article surface using any one of number
of techniques as described herein. After the initiator and natural
biodegradable polysaccharide are both disposed, the initiator is
activated, resulting in the activation of the pendent coupling
groups, the coupling of natural biodegradable polysaccharide
molecules, and the formation of the coating. The steps of disposing
and activating can be performed in ways (as described herein and/or
known in the art) to precisely control the formation of a coating.
For example, the thickness and the location of the coating on the
article surface can be controlled using techniques described herein
and/or known in the art.
[0270] In preferred aspects of the following methods, the natural
biodegradable polysaccharide is selected from the group of amylose
and maltodextrin. In other preferred aspects of the following
methods, the natural biodegradable polysaccharide has a molecular
weight of 500,000 Da or less, 250,000 Da or less, 100,000 Da or
less, or 50,000 Da or less. It is also preferred that the natural
biodegradable polysaccharides have an average molecular weight of
500 Da or greater. A particularly preferred size range for the
natural biodegradable polysaccharides is in the range of about 1000
Da to about 10,000 Da.
[0271] For example, in some aspects the method includes the steps
of (i) disposing a composition comprising (a) a natural
biodegradable polysaccharide having a coupling group, (b) an
initiator, and (c) a bioactive agent on a surface; and (ii)
activating the initiator to provide a coated composition having the
natural biodegradable polysaccharide and the bioactive agent on the
surface. This method can also be used to form a medical implant
wherein the composition is disposed to form an implant of a desired
configuration. For example, the composition can be disposed in a
mold. This method can also be used to form an in situ formed matrix
wherein the composition is disposed within a portion of a
subject.
[0272] In other aspects, the method includes the steps of (i)
disposing an initiator on a surface, (ii) disposing a composition
comprising (a) a natural biodegradable polysaccharide having a
coupling group and (b) a bioactive agent on the surface; and (iii)
activating the initiator to provide a coated composition having the
natural biodegradable polysaccharide and the bioactive agent.
[0273] During the step of activating, a composition including the
natural biodegradable polysaccharide and the bioactive agent are
contacted with the initiator and the initiator is activated to
promote the crosslinking of two or more natural biodegradable
polysaccharides via their coupling groups. In preferred aspects the
natural biodegradable polysaccharide includes a polymerizable
group, such as an ethylenically unsaturated group, and initiator is
capable of initiating free radical polymerization of the
polymerizable groups. Therefore, in another embodiment, the
invention provides a method for coating a surface, including the
steps of (i) disposing a composition comprising (a) a natural
biodegradable polysaccharide having a ethylenically unsaturated
group, (b) a polymerization initiator, and (c) a bioactive agent on
a surface; and (ii) activating the polymerization initiator to
cause the polymerization of the amylose compound thereby providing
a coated composition having the natural biodegradable
polysaccharide and the bioactive agent on the surface. These
methods can also be used to form medical implants and in
situ-formed matrices, wherein the composition is disposed in a mold
or in a subject, respectively, rather than on a surface.
[0274] In yet another aspect the invention provides a medical
device having a coated composition comprising a plurality of
coupled natural biodegradable polysaccharide and a bioactive
agent.
[0275] In some embodiments, the invention provides methods for
preparing biodegradable coatings that include (a) a natural
biodegradable polysaccharide having a coupling group and (b)
biodegradable microparticles having a bioactive agent.
[0276] In some embodiments the coupling group can be activated by
an initiator. Therefore, the method can include the steps of (i)
disposing an initiator on a surface, (ii) disposing a composition
comprising (a) a natural biodegradable polysaccharide having a
coupling group and (b) biodegradable microparticles comprising a
bioactive agent; and (iii) activating the initiator to provide a
biodegradable bioactive agent-releasing coated composition having
the natural biodegradable polysaccharide and the biodegradable
microparticles having the bioactive agent.
[0277] In preferred aspects the natural biodegradable
polysaccharide includes a polymerizable group, such as an
ethylenically unsaturated group, and initiator is capable of
initiating free radical polymerization of the polymerizable groups.
Therefore, in another embodiment, the invention provides a method
for coating a surface, including the steps of (i) disposing a
composition comprising (a) a natural biodegradable polysaccharide
having an ethylenically unsaturated group, (b) a polymerization
initiator, and (c) biodegradable microparticles having a bioactive
agent on a surface; and (ii) activating the polymerization
initiator to cause the polymerization of the natural biodegradable
polysaccharide thereby providing a coated composition that includes
biodegradable microparticles in a natural biodegradable
polysaccharide matrix.
[0278] The invention also provides alternative methods for
preparing a coated surface that is biodegradable and having
microparticles that can release a bioactive agent. The methods
include disposing in two or more steps at least the following
reagents on a surface: (a) a natural biodegradable polysaccharide
comprising a first coupling group (b) a natural biodegradable
polysaccharide comprising a second coupling group that is reactive
with the first coupling group, and (c) biodegradable microparticles
that include a bioactive agent. According to this method reagents
(a) and (b) are reactive with each other and are disposed
separately on the surface but can individually include (c). For
example, reagent (a) is first disposed on the surface and then a
mixture comprising reagent (b) and (c) is then disposed on reagent
(a). Reagent (a) reacts with (b) to link the natural biodegradable
polysaccharide together to form a coating that includes (c), the
biodegradable microparticles.
[0279] The invention also provides methods for preparing
biodegradable sealant coatings that include a natural biodegradable
polysaccharide having a coupling group; optionally a bioactive
agent can be included in the sealant coating.
[0280] In some embodiments, the method includes the steps of (i)
disposing a sealant composition comprising (a) a natural
biodegradable polysaccharide having a coupling group, and (b) an
initiator, and (ii) activating the initiator to form a sealant
coating. This aspect of the invention includes coating methods
where a bulk polymerization approach is performed. For example, in
some embodiments, a composition including a polymerization
initiator and natural biodegradable polysaccharides having a
polymerizable group is disposed on a surface. The initiator is then
activated to promote bulk polymerization and coupling of the
natural biodegradable polysaccharides in association with the
surface.
[0281] In other aspects, the method includes the steps of (i)
disposing an initiator on a surface, (ii) disposing a natural
biodegradable polysaccharide having a coupling group; and (iii)
activating the initiator to provide a coated composition having the
amylose polymer. The natural biodegradable polysaccharides can be
disposed on the surface along with other reagents if desired. This
aspect of the invention includes coating methods where a graft
polymerization approach is performed. For example, in some
embodiments, a polymerization initiator is first disposed on a
surface and then a natural biodegradable polysaccharide having a
polymerizable group is disposed on the surface having the
initiator. The initiator is activated to promote free radical
polymerization, and coupling of the natural biodegradable
polysaccharides from the surface.
[0282] In other embodiments of the invention, an aqueous
composition that includes the natural biodegradable polysaccharide
having the coupling group and a bioactive agent is obtained and
used in the method of providing a sealant coating to a surface.
This composition can be prepared by mixing the natural
biodegradable polysaccharide with a bioactive agent, for example, a
water-soluble small molecule, a protein, or a nucleic acid. In one
preferred aspect of the invention, the bioactive agent is a
procoagulant or prothrombotic factor. For example, the bioactive
agent can be a protein such as recombinant collagen, or other
proteins that associate with receptors on platelets to induce
platelet aggregation.
[0283] In some aspects of the invention, the coating is placed in
contact with an aqueous solution, or the materials of the coating
composition. The coating or coating materials are designed to be
stable in the presence of the aqueous solution provided that an
enzyme that causes the degradation of the natural biodegradable
polysaccharide (or another degrading agent) is not present in an
amount sufficient to cause substantial degradation of the
materials.
[0284] For example, the invention provides a shelf stable
composition comprising a natural biodegradable polysaccharide
comprising coupling groups. These compositions could be obtained or
prepared, according to the details provided herein, and then stored
for a period of time before the composition is used to form a
biodegradable coating, without the significant degradation of the
natural biodegradable polysaccharide occurring during storage.
[0285] Accordingly, the invention also provides methods for
preparing a biodegradable coating comprising preparing a
biodegradable coating composition comprising a natural
biodegradable polysaccharide comprising coupling group; storing the
coating composition for an amount of time; and then using the
coating composition to prepare a biodegradable coating. Optionally,
one or more bioactive agents and/or microparticles can be added
before or after storage of the coating composition.
[0286] In a related aspect, the invention also provides the
advantage of being able to perform synthetic and post-synthetic
procedures wherein the natural biodegradable polysaccharide is
contacted with an aqueous composition, and there is minimal risk of
degradation of the polysaccharide. For example, the natural
biodegradable polysaccharide may be contacted with an aqueous
solution for purification without risking significant degradation
of the natural biodegradable polysaccharide.
[0287] In yet another aspect, the invention relates to the
stability of the coatings that are formed on an article. The
invention provides a method comprising obtaining an article having
a coating comprising a natural biodegradable polysaccharide, and
then contacting the article with an aqueous solution. The aqueous
solution can be, for example, a storage solution, a solution that
is used to hydrate the surface of the coated device, or an aqueous
sterilization solution.
[0288] In some aspects the coating can be contacted with an aqueous
sterilization solution. Medical articles, or parts of medical
articles, can be prepared having a coating and these articles can
be treated to sterilize one or more parts of the article, or the
entire medical article. Sterilization can take place prior to using
the medical article and/or, in some cases, during implantation of
the medical article.
[0289] In some aspects, the invention provides a method for
delivering a bioactive agent from a biodegradable matrix by
exposing the matrix to an enzyme that causes its degradation. The
matrix can be in the form of a coating on the surface of an
implantable article, or can provide the structure of an implantable
article. In performing this method an article having a
biodegradable matrix comprising a natural biodegradable material is
provided to a subject. The article is then exposed to an enzyme,
such as a carbohydrase, that can promote the degradation of the
matrix. The structure of the matrix prevents the enzyme from
entering the matrix, and degradation of the matrix occurs at its
surface. Bioactive agent is released from the surface as the matrix
is degraded. Release of the bioactive agent is observed in a zero
order rate, meaning that the amount of bioactive agent that is
released from the matrix per time remains essentially constant
throughout the lifespan of the matrix.
[0290] In some aspects, the enzyme degrades a biodegradable coating
comprising a natural biodegradable polysaccharide having pendent
coupling groups, wherein the coating is formed on a surface of the
article by reaction of the coupling groups to form a crosslinked
matrix of a plurality of natural biodegradable polysaccharides, and
wherein the coating includes a bioactive agent.
[0291] In these aspects, the carbohydrase that contacts the coating
or article can specifically degrade the natural biodegradable
polysaccharide causing release of the bioactive agent. Examples of
carbohydrases that can specifically degrade natural biodegradable
polysaccharide coatings include .alpha.-amylases, such as salivary
and pancreatic .alpha.-amylases; disaccharidases, such as maltase,
lactase and sucrase; trisaccharidases; and glucoamylase
(amyloglucosidase).
[0292] Serum concentrations for amylase are estimated to be in the
range of about 50-100 Upper liter, and vitreal concentrations also
fall within this range (Varela, R. A., and Bossart, G. D. (2005) J
Am Vet Med Assoc 226:88-92).
[0293] A method for causing the degradation of the matrix can also
include a step of providing an enzyme that promotes the degradation
of the matrix. The enzyme can be provided by, for example,
administering the enzyme to a subject, or associating the enzyme
with a portion of the implanted article, or another article placed
within the subject.
[0294] For example, in some aspects, a carbohydrase is be
administered to a subject to increase the local concentration, for
example in the serum or the tissue surrounding the implanted
device, so that the carbohydrase may promote the degradation of the
coating. Exemplary routes for introducing a carbohydrase include
local injection, intravenous (IV) routes, and the like.
Alternatively, degradation can be promoted by indirectly increasing
the concentration of a carbohydrase in the vicinity of the coated
article, for example, by a dietary process, or by ingesting or
administering a compound that increases the systemic levels of a
carbohydrase.
[0295] In other cases, the carbohydrase can be provided on a
portion of the coated article. For example the carbohydrase may be
eluted from a portion of the article that does not have the natural
biodegradable polymer coating. In this aspect, as the carbohydrase
is released it locally acts upon the coating to cause its
degradation and promote the release of the bioactive agent.
Alternatively, the carbohydrase can be present in a microparticle
in one or more portions the coating. As the carbohydrase is
released from the microparticle, it causes coating degradation and
promote the release of the bioactive agent.
[0296] In another aspect, the invention sets forth methods for
providing lubricity to an article surface. The matrix of
biodegradable polysaccharides as described herein can provide a
surprisingly lubricious and durable surface when formed, for
example, on the surface of a device.
[0297] As used herein, the term "lubricity" refers to a
characterization of the frictional force associated with a coating.
A lubricious coating can reduce the frictional forces present on
the surface of the device when another surface is moved against the
device surface. For example, a catheter having a coating that
provides improved lubricity will encounter less frictional
resistance when moved within a portion of the body, as compared to
an uncoated substrate, or a coating that is not lubricious.
Lubricity can also be important for devices with inner moving parts
in addition to devices that function along with another device, for
example, a coronary catheter which guides the insertion of a PTCA
catheter. The methods can be used to prepare lubricious coatings
for short term use and/or single use devices.
[0298] Improved lubricity can be shown by one or more methods. One
method of testing lubricity of a coating is by the horizontal sled
style friction test method (such as ASTM D-1894; a modified test is
described herein). The lubricity measurements described herein
refer to the kinetic coefficient of friction, which is equal to the
average force reading obtained during uniform sliding of the
surfaces divided by the sled weight. The measurements are in
grams.
[0299] As shown herein, the biodegradable polysaccharide coatings
generally show a lubricity of 20 g or less, and coatings having a
lubricity of 15 g or less, or 10 g or less can be prepared by the
methods described herein. In one desirable method for preparing the
coating, a biodegradable polysaccharide matrix is formed using a
photoinitiator to promote association of the biodegradable
polysaccharides and matrix formation.
[0300] Another method which can be used to demonstrate an
improvement in lubricity is the water contact angle measurement
method. Reduction of water contact angle is indicative of increased
wettability, which associates with an improvement in lubricity.
[0301] Also, in many aspects, the biodegradable coating of the
invention demonstrates excellent durability. As used herein, the
term "durability" refers to the wear resistance of the
biodegradable polysaccharide coating. A more durable coating is
less easily removed from a substrate by abrasion. Durability of a
coating can be assessed by subjecting the device to conditions that
simulate use conditions. The durability of the coating compositions
is demonstrated by the ability to adhere to the device surface
sufficiently to withstand the effect of shear forces encountered
during insertion and/or removal of the device, which could
otherwise result in delamination of the coating from the body
member.
[0302] Improved durability can be shown by one or more methods. One
preferred method of testing durability, as well as lubricity, is by
the horizontal sled style friction test method described herein. In
the method multiple push-pull cycles are performed and friction
measurements are taken during the measurements. If minimal increase
in friction values is seen after multiple push pull cycles, the
coating is though to have good durability.
[0303] The biodegradable coatings of the present invention showed
excellent durability. For example, the lubricity of the fifth push
and pull cycle generally was not greater than 10% or more typically
not greater than 5% of the lubricity of the first push and pull
cycle. At later points, for example, at the fortieth push and pull
cycle, the lubricity generally was not greater than 20% or more
typically not greater than 10% of the lubricity of the first push
and pull cycle.
[0304] The invention will be further described with reference to
the following non-limiting Examples. It will be apparent to those
skilled in the art that many changes can be made in the embodiments
described without departing from the scope of the present
invention. Thus the scope of the present invention should not be
limited to the embodiments described in this application, but only
by embodiments described by the language of the claims and the
equivalents of those embodiments. Unless otherwise indicated, all
percentages are by weight.
Example 1
Synthesis of Acrylated-Amylose
[0305] Amylose having polymerizable vinyl groups was prepared by
mixing 0.75 g of amylose (A0512; Aldrich) with 100 mL of
methylsulfoxide (JT Baker) in a 250 mL amber vial, with stirring.
After one hour, 2 mL of triethylamine (TEA; Aldrich) was added and
the mixture was allowed to stir for 5 minutes at room temperature.
Subsequently, 2 mL of glycidyl acrylate (Polysciences) was added
and the amylose and glycidyl acrylate were allowed to react by
stirring overnight at room temperature. The mixture containing the
amylose-glycidyl acrylate reaction product was dialyzed for 3 days
against DI water using continuous flow dialysis. The resultant
acrylated-amylose (0.50 g; 71.4% yield) was then lyophilized and
stored desiccated at room temperature with protection from
light.
Example 2
Synthesis of MTA-PAAm
[0306] A polymerization initiator was prepared by copolymerizing a
methacrylamide having a photoreactive group with acrylamide.
[0307] A methacrylamide-oxothioxanthene monomer
(N-[3-(7-Methyl-9-oxothioxanthene-3-carboxamido)
propyl]methacrylamide (MTA-APMA)) was first prepared.
N-(3-aminopropyl)methacrylamide hydrochloride (APMA), 4.53 g (25.4
mmol), prepared as described in U.S. Pat. No. 5,858,653, Example 2,
was suspended in 100 mL of anhydrous chloroform in a 250 mL round
bottom flask equipped with a drying tube.
7-methyl-9-oxothioxanthene-3-carboxylic acid (MTA) was prepared as
described in U.S. Pat. No. 4,506,083, Example D. MTA-chloride
(MTA-Cl) was made as described in U.S. Pat. No. 6,007,833, Example
1. After cooling the slurry in an ice bath, MTA-Cl (7.69 g; 26.6
mmol) was added as a solid with stirring to the APMA-chloroform
suspension. A solution of 7.42 mL (53.2 mmol) of TEA in 20 mL of
chloroform was then added over a 1.5 hour time period, followed by
a slow warming to room temperature. The mixture was allowed to stir
16 hours at room temperature under a drying tube. After this time,
the reaction was washed with 0.1 N HCl and the solvent was removed
under vacuum after adding a small amount of phenothiazine as an
inhibitor. The resulting product was recrystallized from
tetrahydrofuran (THF)/toluene (3/1) and gave 8.87 g (88.7% yield)
of product after air drying. The structure of MTA-APMA was
confirmed by NMR analysis.
[0308] MTA-APMA was then copolymerized with acrylamide in DMSO in
the presence of 2-mercaptoethanol (a chain transfer agent),
N,N,N',N'-tetramethyl-ethylenediamine (a co-catalyst), and
2,2'-azobis(2-methyl-propionitrile) (a free radical initiator) at
room temperature. The solution was sparged with nitrogen for 20
minutes, sealed tightly, and incubated at 55.degree. C. for 20
hours. The solution was dialyzed for 3 days against DI water using
continuous flow dialysis. The resultant MTA-PAAm was lyophilized,
stored desiccated, and protected from light at room
temperature.
Example 3
Formation of an Amylose Coating
[0309] 100 mg of acrylated-amylose as prepared in Example 1 was
placed in an 8 mL amber vial. To the acrylated-amylose was added 3
mg of MTA-PAAm (lyophilized), 2 of 2-NVP (N-vinyl-2-pyrrolidone;
accelerant (Bimax)) and 1 mL of 1.times. phosphate-buffered saline
(1.times.PBS). The reagents were then mixed for one hour on a
shaker at 37.degree. C. The mixture in an amount of 50 .mu.L was
placed onto a glass slide (2991FI; Esco) and illuminated for 50
seconds with an EFOS 100 SS illumination system equipped with a
400-500 nm filter (50 mW/cm.sup.2). After illumination the polymer
was found to form a semi-firm gel having elastomeric
properties.
Example 4
Preparation of 4-bromomethylbenzophenone (BMBP)
[0310] 4-Methylbenzophenone (750 g; 3.82 moles) was added to a 5
liter Morton flask equipped with an overhead stirrer and dissolved
in 2850 mL of benzene. The solution was then heated to reflux,
followed by the dropwise addition of 610 g (3.82 moles) of bromine
in 330 mL of benzene. The addition rate was approximately 1.5
mL/min and the flask was illuminated with a 90 watt (90 joule/sec)
halogen spotlight to initiate the reaction. A timer was used with
the lamp to provide a 10% duty cycle (on 5 seconds, off 40
seconds), followed in one hour by a 20% duty cycle (on 10 seconds,
off 40 seconds). At the end of the addition, the product was
analyzed by gas chromatography and was found to contain 71% of the
desired 4-bromomethylbenzophenone, 8% of the dibromo product, and
20% unreacted 4-methylbenzophenone. After cooling, the reaction
mixture was washed with 10 g of sodium bisulfite in 100 mL of
water, followed by washing with 3.times.200 mL of water. The
product was dried over sodium sulfate and recrystallized twice from
1:3 toluene:hexane. After drying under vacuum, 635 g of
4-bromomethylbenzophenone was isolated, providing a yield of 60%,
having a melting point of 112.degree. C.-114.degree. C. Nuclear
magnetic resonance ("NMR") analysis (.sup.1H NMR (CDCl.sub.3)) was
consistent with the desired product: aromatic protons 7.20-7.80 (m,
9H) and methylene protons 4.48 (s, 2H). All chemical shift values
are in ppm downfield from a tetramethylsilane internal
standard.
Example 5
Preparation of ethylenebis(4-benzoylbenzyldimethylammonium)
dibromide
[0311] N,N,N',N'-Tetramethylethylenediamine (6 g; 51.7 mmol) was
dissolved in 225 mL of chloroform with stirring. BMBP (29.15 g;
106.0 mmol), as described in Example 4, was added as a solid and
the reaction mixture was stirred at room temperature for 72 hours.
After this time, the resulting solid was isolated by filtration and
the white solid was rinsed with cold chloroform. The residual
solvent was removed under vacuum and 34.4 g of solid was isolated
for a 99.7% yield, melting point 218.degree. C.-220.degree. C.
Analysis on an NMR spectrometer was consistent with the desired
product: .sup.1H NMR (DMSO-d.sub.6) aromatic protons 7.20-7.80 (m,
18H), benzylic methylenes 4.80 (br. s, 4H), amine methylenes 4.15
(br. s, 4H), and methyls 3.15 (br. s, 12H).
Example 6
Formation of an Amylose Matrix on PET Mesh
[0312] Acrylated-amylose (100 mg), as described in Example 1, was
placed in an 8 mL amber vial.
Ethylenebis(4-benzoylbenzyldimethylammonium) dibromide (3 mg), as
described in Example 5, 2 .mu.l of 2-NVP, and 1 mL of 1.times.
phosphate buffered saline (1.times.PBS) was added to the
acrylated-amylose and mixed for two hours on a shaker at 37.degree.
C. The mixture (250 .mu.l) was spread onto a 3 cm.times.2 cm
polyethylene terephthalate (PET) mesh substrate (41 .mu.m monofil
diameter; Goodfellow Cambridge Ltd., UK). The PET substrate with
the applied amylose mixture was placed in a Dymax Lightweld PC-2
illumination system (Dymax Corp.; light intensity 6.5 mW/cm.sup.2),
15 cm from the light source, and illuminated for 60 seconds. After
illumination, the applied amylose mixture was found to form a
semi-firm gel on the PET substrate, with elastomeric properties
evident.
Example 7
Preparation of 1-(6-oxo-6-hydroxyhexyl)maleimide (Mal-EACA)
[0313] A maleimide functional acid was prepared in the following
manner, and was used in Example 8. EACA (6-aminocaproic acid), (100
g; 0.762 moles), was dissolved in 300 mL of acetic acid in a
three-neck, three liter flask equipped with an overhead stirrer and
drying tube. Maleic anhydride, (78.5 g; 0.801 moles), was dissolved
in 200 mL of acetic acid and added to the EACA solution. The
mixture was stirred one hour while heating on a boiling water bath,
resulting in the formation of a white solid. After cooling
overnight at room temperature, the solid was collected by
filtration and rinsed two times with 50 mL of hexane each rinse.
After drying, the yield of the (z)-4-oxo-5-aza-undec-2-endioic acid
(Compound 1) was in the range of 158-165 g (90-95%) with a melting
point of 160-165.degree. C. Analysis on an NMR spectrometer was
consistent with the desired product: .sup.1H NMR (DMSO-d.sub.6, 400
MHz) .delta. 6.41, 6.24 (d, 2H, J=12.6 Hz; vinyl protons), 3.6-3.2
(b, 1H; amide proton), 3.20-3.14 (m, 2H: methylene adjacent to
nitrogen), 2.20 (t, 2H, J=7.3; methylene adjacent to carbonyl),
1.53-1.44 (m, 4H; methylenes adjacent to the central methylene),
and 1.32-1.26 (m, 2H; the central methylene).
[0314] (z)-4-oxo-5-aza-undec-2-endioic acid, (160 g; 0.698 moles),
zinc chloride, 280 g (2.05 moles), and phenothiazine, 0.15 g were
added to a two liter round bottom flask fitted with an overhead
stirrer, condenser, thermocouple, addition funnel, an inert gas
inlet, and heating mantle. Chloroform (CHCl.sub.3), 320 mL was
added to the 2 liter reaction flask, and stirring of the mixture
was started. Triethylamine (480 mL; 348 g, 3.44 moles (TEA)) was
added over one hour. Chlorotrimethyl silane (600 mL; 510 g, 4.69
moles) was then added over two hours. The reaction was brought to
reflux and was refluxed overnight (-16 hours). The reaction was
cooled and added to a mixture of CHCl.sub.3 (500 mL), water (1.0
liters), ice (300 g), and 12 N hydrochloric acid (240 mL) in a 20
liter container over 15 minutes. After 15 minutes of stirring, the
aqueous layer was tested to make sure the pH was less than 5. The
organic layer was separated, and the aqueous layer was extracted
three times with CHCl.sub.3 (700 mL) each extraction. The organic
layers were combined and evaporated on a rotary evaporator. The
residue was then placed in a 20 liter container. A solution of
sodium bicarbonate (192 g) in water (2.4 liters) was added to the
residue. The bicarbonate solution was stirred until the solids were
dissolved. The bicarbonate solution was treated with a solution of
hydrochloric acid, (26 liters of 1.1 N) over 5 minutes to a pH of
below 2. The acidified mixture was then extracted with two portions
of CHCl.sub.3, (1.2 liters and 0.8 liters) each extraction. The
combined extracts were dried over sodium sulfate and evaporated.
The residue was recrystallized from toluene and hexane. The
crystalline product was then isolated by filtration and dried which
produced 85.6 g of white N-(6-oxo-6-hydroxyhexyl)maleimide
(Mal-EACA; Compound 2). Analysis on an NMR spectrometer was
consistent with the desired product: .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 6.72 (s, 2H; maleimide protons), 3.52 (t, 2H, J=7.2
Hz; methylene next to maleimide), 2.35 (t, 2H, J=7.4; methylene
next to carbonyl), 1.69-1.57 (m, 4H; methylenes adjacent to central
methylene), and 1.39-1.30 (m, 2H; the central methylene). The
product had a DSC (differential scanning calorimator) melting point
peak at 89.9.degree. C.
##STR00001##
Example 8
Preparation of N-(5-isocyanatopentyl)maleimide (Mal-C5-NCO)
[0315] Mal-EACA from Example 7 (5.0 g; 23.5 mmole) and CHCl.sub.3
(25 mL) were placed in a 100 mL round bottom flask and stirred
using a magnetic bar with cooling in an ice bath. Oxalyl chloride
(10.3 mL; .about.15 g; 118 mmole) was added and the reaction was
brought to room temperature with stirring overnight. The volatiles
were removed on a rotary evaporator, and the residue was azetroped
with three times with 10 mL CHCl.sub.3 each time. The intermediate
Mal-EAC-Cl [N-(6-oxo-6-chlorohexyl)maleimide] (Compound 3) was
dissolved in acetone (10 mL) and added to a cold (ice bath) stirred
solution of sodium azide (2.23 g; 34.3 mmole) in water (10 mL). The
mixture was stirred one hour using an ice bath. The organic layer
was set aside in an ice bath, and the aqueous layer was extracted
three times with 10 mL CHCl.sub.3. All operations of the acylazide
were done at ice bath temperatures. The combined organic solutions
of the azide reaction were dried for an hour over anhydrous sodium
sulfate. The N-(6-oxo-6-azidohexyl)maleimide (Compound 4) solution
was further dried by gentle swirling over molecular sieves over
night. The cold azide solution was filtered and added to refluxing
CHCl.sub.3, 5 mL over a 10 minute period. The azide solution was
refluxed for 2 hours. The weight of Mal-C5-NCO (Compound 5)
solution obtained was 55.5 g, which was protected from moisture. A
sample of the isocyanate solution, 136 mg was evaporated and
treated with DBB (1,4-dibromobenzene), 7.54 mg and chloroform-d,
0.9 mL: .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 6.72 (s, 2H),
3.55 (t, 2H, J=7.2 Hz), 3.32 (t, 2H, J=6.6 Hz), 1.70-1.59 (m, 4H),
1.44-1.35 (m, 2H). The NMR spectra was consistent with desired
product. The DBB internal standard 8 at 7.38 (integral value was
2.0, 4H; per mole of product) was used to estimate the moles of
Mal-C5-NCO in solution. The calculated amount of product in
solution was 23.2 mmole for a yield of 98% of theory. NCO reagent
(concentration was 0.42 mmole/g) was used to prepare a macromer in
Example 14.
##STR00002##
Example 9
Preparation of 3-(acryloyloxy)propanoic acid (2-carboxyethyl
acrylate; CEA)
[0316] Acrylic acid (100 g; 1.39 mole) and phenothiazine (0.1 g)
were placed in a 500 mL round bottom flask. The reaction was
stirred at 92.degree. C. for 14 hours. The excess acrylic acid was
removed on a rotary evaporator at 25.degree. C. using a mechanical
vacuum pump. The amount of residue obtained was 51.3 g. The CEA
(Compound 6) was used in Example 10 without purification.
##STR00003##
Example 10
Preparation of 3-chloro-3-oxopropyl acrylate (CEA-Cl)
[0317] CEA from Example 9 (51 g; 0.35 mole) and dimethyl formamide
(DMF; 0.2 mL; 0.26 mmole) were dissolved in CH.sub.2Cl.sub.3 (100
mL). The CEA solution was added slowly (over 2 hours) to a stirred
solution of oxalyl chloride (53 mL; 0.61 mole), DMF (0.2 mL; 2.6
mmole), anthraquinone (0.5 g; 2.4 mmole), phenothiazine (0.1 g, 0.5
mmole), and CH.sub.2Cl.sub.3 (75 mL) in a 500 mL round bottom flask
in an ice bath at 200 mm pressure. A dry ice condenser was used to
retain the CH.sub.2Cl.sub.3 in the reaction flask. After the
addition was complete the reaction was stirred at room temperature
overnight. The weight of reaction solution was 369 g. A sample of
the CEA-Cl (Compound 7) reaction solution (124 mg) was treated with
1,4-dibromobenzene (DBB, 6.85 mg) evaporated and dissolved in
CDCl.sub.3: .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.38 (s, 4H;
DBB internal std.), 6.45 (d, 1H, J=17.4 Hz), 6.13 (dd, 1H, J=17.4,
10.4 Hz), 5.90 (d, 1H, J=10.4 Hz), 4.47 (t, 2H, J=5.9 Hz), 3.28 (t,
2H, J=5.9). The spectra was consistent with the desired product.
There was 0.394 mole DBB for 1.0 mole CEA-Cl by integration, which
gave a calculated yield of 61%. Commercially available CEA (426 g;
Aldrich) was reacted with oxalyl chloride (532 mL) in a procedure
similar to the one listed above. The residue CEA-Cl (490 g) was
distilled using an oil bath at 140.degree. C. at a pressure of 18
mm Hg. The distillate temperature reached 98.degree. C. and 150 g
of distillate was collected. The distillate was redistilled at 18
mm Hg at a maximum bath temperature of 120.degree. C. The
temperature range for the distillate was 30.degree. C. to
70.degree. C. which gave 11 g of material. The distillate appeared
to be 3-chloro-3-oxopropyl 3-chloropropanoate. The residue of the
second distillation (125 g; 26% of theory) was used in Example
11.
##STR00004##
Example 11
Preparation of 3-azido-3-oxopropyl acrylate (CEA-N3)
[0318] CEA-Cl from Example 10 (109.2 g; 0.671 mole) was dissolved
in acetone (135 mL). Sodium azide (57.2 g; 0.806 mole) was
dissolved in water (135 mL) and chilled. The CEA-Cl solution was
then added to the chilled azide solution with vigorous stirring in
an ice bath for 1.5 hours. The reaction mixture was extracted two
times with 150 mL of CHCl.sub.3 each extraction. The CHCl.sub.3
solution was passed through a silica gel column 40 mm in diameter
by 127 mm. The 3-azido-3-oxopropyl acrylate (Compound 8) solution
was gently agitated over dried molecular sieves at 4.degree. C.
overnight. The dried solution was used in Example 12 without
purification.
##STR00005##
Example 12
Preparation of 2-isocyanatoethyl acrylate (EA-NCO)
[0319] The dried azide solution (from Example 11) was slowly added
to refluxing CHCl.sub.3, 75 mL. After the addition was completed,
refluxing was continued 2 hours. The EA-NCO (Compound 9) solution
(594.3 g) was protected from moisture. A sample of the EA-NCO
solution (283.4 mg) was mixed with DBB (8.6 mg) and evaporated. The
residue was dissolved in CDCl.sub.3: .sup.1H NMR (CDCl.sub.3, 400
MHz) .delta. 7.38 (s, 4H; DBB internal std.), 6.50 (d, 1H, J=17.3
Hz), 6.19 (dd, 1H, J=17.3, 10.5 Hz), 5.93 (d, 1H, J=10.5 Hz), 4.32
(t, 2H, J=5.3 Hz), 3.59 (t, 2H, J=5.3). The spectra was consistent
with the desired EA-NCO. There was 0.165 mole DBB for 1.0 mole
EA-NCO by integration, which gave a calculated concentration of 110
mg EA-NCO/g of solution. The EA-NCO solution was used to prepare a
macromer in Example 13.
##STR00006##
Example 13
Preparation of Maltodextrin-Acrylate Macromer (Md-Acrylate)
[0320] Maltodextrin (MD; Aldrich; 9.64 g; .about.3.21 mmole; DE
(Dextrose Equivalent): 4.0-7.0) was dissolved in dimethylsulfoxide
(DMSO) 60 mL. The size of the maltodextrin was calculated to be in
the range of 2,000 Da-4,000 Da. A solution of EA-NCO from Example
12 (24.73 g; 19.3 mmole) was evaporated and dissolved in dried DMSO
(7.5 mL). The two DMSO solutions were mixed and heated to
55.degree. C. overnight. The DMSO solution was placed in dialysis
tubing (1000 MWCO, 45 mm flat width.times.50 cm long) and dialyzed
against water for 3 days. The macromer solution was filtered and
lyophilized to give 7.91 g white solid. A sample of the macromer
(49 mg), and DBB (4.84 mg) was dissolved in 0.8 mL DMSO-d.sub.6:
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 7.38 (s, 4H; internal
std. integral value of 2.7815), 6.50, 6.19, and 5.93 (doublets, 3H;
vinyl protons integral value of 3.0696). The calculated acrylate
load of macromer was 0.616 .mu.moles/mg of polymer.
Example 14
Preparation of Maltodextrin-Maleimide Macromer (MD-Mal)
[0321] A procedure similar to Example 13 was used to make the
MD-Mal macromer. A solution of Mal-C5-NCO from Example 8 (0.412 g;
1.98 mmole) was evaporated and dissolved in dried DMSO (2 mL). MD
(0.991 g; 0.33 mmole) was dissolved in DMSO (5 mL). The DMSO
solutions were combined and stirred at 55.degree. C. for 16 hours.
Dialysis and lyophilization gave 0.566 g product. A sample of the
macromer (44 mg), and DBB (2.74 mg) was dissolved in 00.8 mL
DMSO-d.sub.6: .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 7.38 (s,
4H; internal std. integral value of 2.3832), 6.9 (s, 2H; Maleimide
protons integral value of 1.000). The calculated acrylate load of
macromer was 0.222 .mu.moles/mg of polymer. The macromer was tested
for its ability to make a matrix (see Example 17)
Example 15
Formation of Maltodextrin-acrylate Biodegradable Matrix Using
MTA-PAAm
[0322] 250 mg of MD-Acrylate as prepared in Example 13 was placed
in an 8 mL amber vial. To the MD-Acrylate was added 3 mg of
MTA-PAAm (lyophilized), 2 .mu.L of 2-NVP, and 1 mL of 1.times.
phosphate-buffered saline (1.times.PBS), providing a composition
having MD-Acrylate at 250 mg/mL. The reagents were then mixed for
one hour on a shaker at 37.degree. C. The mixture in an amount of
50 .mu.L was placed onto a glass slide and illuminated for 40
seconds with an EFOS 100 SS illumination system equipped with a
400-500 nm filter. After illumination the polymer was found to form
a semi-firm gel having elastomeric properties.
Example 16
Formation of MD-Acrylate Biodegradable Matrix Using
Camphorquinone
[0323] 250 mg of MD-acrylate as prepared in Example 13 was placed
in an 8 mL amber vial. To the MD-Acrylate was added 14 mg of
camphorquinone-10-sulfonic acid hydrate (Toronto Research
Chemicals, Inc.), 3 .mu.L of 2-NVP, and 1 mL of distilled water.
The reagents were then mixed for one hour on a shaker at 37.degree.
C. The mixture in an amount of 50 .mu.L, was placed onto a glass
slide and illuminated for 40 seconds with a SmartliteIQ.TM. LED
curing light (Dentsply Caulk). After illumination the polymer was
found to form a semi-firm gel having with elastomeric
properties.
Example 17
Formation of MD-Mal Biodegradable Matrix Using MTA-PAAm
[0324] 250 mg of MD-Mal as prepared in Example 14 was placed in an
8 mL amber vial. To the MD-Mal was added 3 mg of MTA-PAAm
(lyophilized), 2 .mu.L of 2-NVP, and 1 mL of 1.times.
phosphate-buffered saline (1.times.PBS). The reagents were then
mixed for one hour on a shaker at 37.degree. C. The mixture in an
amount of 50 .mu.L was placed onto a glass slide and illuminated
for 40 seconds with an EFOS 100 SS illumination system equipped
with a 400-500 nm filter. After illumination the polymer was found
to form a semi-firm gel having elastomeric properties.
Example 18
Coating a PEBAX.RTM. Rod with MD-Acrylate
[0325] 100 mg photo-derivatized poly(vinylpyrrolidone) (photo-PVP)
as prepared as described in U.S. Pat. No. 5,637,460, and the
photoinitiator tetrakis (4-benzoylbenzyl ether) of pentaerythritol
["tetra-BBE-PET"] (5 mg), prepared as described in U.S. Pat. No.
5,414,075 (Example 1) and commercially available from SurModics,
Inc. (Eden Prairie, Minn.) as PRO1, were mixed with 10 mL isopropyl
alcohol (IPA; Fisher) for 1 minute. The mixture in an amount of 1
mL was placed into a 1.8 mL eppendorf tube (VWR). A 1.2 cm
PEBAX.RTM. rod (Medical Profiles, Inc) was dipped into the solution
for 10 seconds, at a dip rate of 0.1 cm/second, and then removed at
the same rate. The rod was allowed to air dry for 5 minutes. The
rod was placed into a Dymax Lightweld PC-2 illumination system
(Dymax Corp.; light intensity 6.5 mW/cm.sup.2), 30 cm from light
source, illuminated for 180 seconds, and then removed.
[0326] 300 mg of MD-Acrylate, as prepared in Example 13, was placed
in an 8 mL amber vial. To the MD-Acrylate was added
4,5-bis(4-benzoylphenylmethyleneoxy) benzene-1,3-disulfonic acid (5
mg) (DBDS), prepared as described in U.S. Pat. No. 6,278,018
(Example 1) and commercially available from SurModics, Inc. (Eden
Prairie, Minn.) as DBDS, and 1 mL of 1.times. phosphate-buffered
saline (1.times.PBS). The reagents were then mixed for one hour on
a shaker at 37.degree. C. The mixture in an amount of 1 mL was
placed into a 1.8 mL eppendorf tube (VWR). The
photo-PVP/tetra-BBE-PET coated PEBAX.RTM. rod was dipped into the
mixture for 30 seconds, at a dip rate of 0.3 cm/s, and then removed
at the same rate. The rod was immediately placed into a Dymax
Lightweld PC-2 illumination system (Dymax Corp.; light intensity
6.5 mW/cm.sup.2), 30 cm from light source, and illuminated for 180
seconds and then removed.
[0327] The MD-Acrylate coated rod was examined under Scanning
Electron Microscope (SEM; LEO Supra 35 VP); the MD-Acrylate coating
thickness varied from 2.1 .mu.m to 2.5 .mu.m, with an average
coating thickness of 2.3 .mu.m.
Example 19
Bioactive Agent Incorporation/Release from a MD-Acrylate Matrix
[0328] 500 mg of MD-Acrylate as prepared in Example 13 was placed
in an 8 mL amber vial. To the MD-Acrylate was added 3 mg of
MTA-PAAm (lyophilized), 2 .mu.L of 2-NVP, and 1 mL of 1.times.
phosphate-buffered saline (1.times.PBS). The reagents were then
mixed for one hour on a shaker at 37.degree. C. To this mixture was
added either 5 mg 70 kD FITC-Dextran or 5 mg 10 kD FITC-Dextran
(Sigma) and vortexed for 30 seconds. The mixture in an amount of
200 .mu.L was placed into a Teflon well plate (8 mm diameter, 4 mm
deep) and illuminated for 40 seconds with an EFOS 100 SS
illumination system equipped with a 400-500 nm filter. The formed
matrix was loose, and not as well crosslinked as the formed
MD-acrylate matrix in Example 17. After illumination, the matrix
was transferred to a 12 well plate (Falcon) and placed in a well
containing 0.6 mL PBS. At daily intervals for 6 days, 150 .mu.L of
PBS was removed from each well and placed into a 96 well plate. The
remaining 850 .mu.L were removed from the samples, and replaced
with 1 mL fresh PBS. The 96 well plate was analyzed for
FITC-Dextran on a spectrophotometer (Shimadzu) at 490 absorbance.
Results showed that at least 70% of the detectable 10 kd or 70 kD
FITC-Dextran was released from the matrix after 2 days. Visual
observation showed that an unquantified amount of 10 kD or 70 kD
FITC-Dextran remained within the matrix after 6 days.
Example 20
Enzyme Degradation of a MD-Acrylate Matrix
[0329] A MD-Acrylate-coated PEBAX rod (from Example 18) was placed
in 5 mL of 1.times. phosphate-buffered saline (PBS) containing 24
.mu.g alpha-Amylase (Sigma; catalog #A6814) for 7 days on a
rotating plate at 37.degree. C. After 7 days, the rod was removed
from the PBS and washed with distilled water. The rod was then
examined under a Scanning Electron Microscope (LEO Supra 35 VP);
upon examination, no trace of the MD-Acrylate coating was detected.
As a control, a MD-Acrylate-coated PEBAX was placed in 1.times.
phosphate-buffered saline (PBS) without alpha-Amylase; upon
examination, the MD-Acrylate coating was intact and showed no signs
of degradation.
Example 21
Polyalditol-acrylate Synthesis
[0330] Polyalditol (PA; GPC; 9.64 g; .about.3.21 mmole) was
dissolved in dimethylsulfoxide (DMSO) 60 mL. The size of the
polyalditol was calculated to be in the range of 2,000 Da-4,000 Da.
A solution of EA-NCO from Example 12 (24.73 g; 19.3 mmole) was
evaporated and dissolved in dried DMSO (7.5 mL). The two DMSO
solutions were mixed and heated to 55.degree. C. overnight. The
DMSO solution was placed in dialysis tubing (1000 MWCO, 45 mm flat
width.times.50 cm long) and dialyzed against water for 3 days. The
polyalditol macromer solution was filtered and lyophilized to give
7.91 g white solid. A sample of the macromer (49 mg), and DBB (4.84
mg) was dissolved in 0.8 mL DMSO-d.sub.6: .sup.1H NMR
(DMSO-d.sub.6, 400 MHz) .delta. 7.38 (s, 4H; internal std. integral
value of 2.7815), 6.50, 6.19, and 5.93 (doublets, 3H; vinyl protons
integral value of 3.0696). The calculated acrylate load of macromer
was 0.616 .mu.moles/mg of polymer.
Example 22
Maltodextrin-acrylate Filaments
[0331] 1,100 milligrams of MD-Acrylate as prepared in Example 13
was placed in an 8 mL amber vial. To the MD-Acrylate was added 1 mg
of a photoinitiator 4,5-bis(4-benzoylphenyl-methyleneoxy)
benzene-1,3-disulfonic acid (5 mg) (DBDS) and 1 mL of 1.times.
phosphate-buffered saline (PBS). The reagents were then mixed for
one hour on a shaker at 37.degree. C. The mixture in an amount of
10 uL was injected, using a 23 gauge needle, into a 22 mm length
opaque silicone tube (P/N 10-447-01; Helix Medical, Carpinteria,
Calif.). The tubing was placed into a Dymax Lightweld PC-2
illumination system (Dymax Corp.; light intensity 6.5 mW/cm.sup.2),
15 cm from light source, illuminated for 270 seconds, and then
removed. After illumination, the filament was removed from the
silicone tubing by rolling a pencil over the tubing, starting from
the back. The filament was firm, which indicated complete
polymerization of the MD-Acrylate. No excess liquid was observed.
The filament was manipulated with forceps. Maltodextrin filaments
were also made from a MD-acrylate solution having concentration of
200 mg/mL. These are physically firm and same as 1,100 mg/ml.
Example 23
Polyalditol-acrylate Filaments
[0332] 1,500 milligrams of polyalditol-acrylate as prepared in
Example 21 was placed in an 8 ml amber vial. To the
polyalditol-acrylate was added 1 mg of DBDS (lyophilized), 15 mg
Bovine Serum Albumin, and 200 uL of 1.times. phosphate-buffered
saline (PBS). The reagents were then mixed for one hour on a shaker
at 37.degree. C. The mixture in an amount of 10 uL was injected,
using a 23 gauge needle, into a 22 mm length opaque silicone tube
(P/N 10-447-01; Helix Medical, Carpinteria, Calif.). The tubing was
placed into a Dymax Lightweld PC-2 illumination system (Dymax
Corp.; light intensity 6.5 mW/cm.sup.2), 15 cm from light source,
illuminated for 270 seconds, and then removed. After illumination,
the filament was removed from the silicone tubing by rolling a
pencil over the tubing, starting from the back. The filament was
firm, which indicated complete polymerization of the
polyalditol-acrylate. No excess liquid was observed. The filament
was manipulated with forceps
Example 24
Amylase Degradation of Maltodextrin-acrylate Filaments
[0333] Maltodextrin-acrylate filaments were synthesized using 200
mg/mL and 1100 mg/mL MD-acrylate as described in example 22 and
were tested for degradation in Amylase solutions. These filaments
were placed in microcentrifuge tubes containing 1 mL of either
1.times.PBS (control), 1.times.PBS containing alpha-Amylase at
0.121 .mu.g/mL (Sigma; catalog #A6814), or 1.times.PBS containing
alpha-Amylase at 24 .mu.g/mL. The tubes were then placed in an
incubator at 37.degree. C.
[0334] After 2 days in the PBS with the 0.121 .mu.g/mL
alpha-Amylase solution the 200 mg/mL filament was completely
degraded, and no trace of the filament was observable. The 200
mg/mL filament in PBS (control) showed no signs of degradation.
[0335] After 33 days in the 1.times.PBS containing alpha-Amylase at
0.121 .mu.g/mL, the 1100 mg/mL filament had lost some of its
initial firmness (as noted by the slightly curled appearance of the
filament), but was still completely intact. The 1,100 mg/mL
filament in the PBS with 24 ug Amylase had completely degraded
after 48 hours. The 1,100 mg/ml filament in the PBS showed no signs
of degradation.
Example 25
Maltodextrin-acrylate Filaments with Bioactive Agent and
Release
[0336] MD-Acrylate in an amount of 1,100 milligrams of as prepared
in Example 13 was placed in an 8 ml amber vial. To the MD-Acrylate
was added 1 mg of DBDS (lyophilized), 15 mg Bovine Serum Albumin
(representing the bioactive agent; and 1 mL of 1.times.
phosphate-buffered saline (1.times.PBS). The reagents were then
mixed for one hour on a shaker at 37.degree. C. The mixture in an
amount of 10 uL was injected, using a 23 gauge needle, into a 22 mm
length opaque silicone tube (P/N 10-447-01; Helix Medical,
Carpinteria, Calif.). The tubing was placed into a Dymax Lightweld
PC-2 illumination system (Dymax Corp.; light intensity 6.5
mW/cm.sup.2), 15 cm from light source, illuminated for 270 seconds,
and then removed. After illumination, the filament was removed from
the silicone tubing by rolling a pencil over the tubing, starting
from the back. The filament was firm, which indicated complete
polymerization of the MD-Acrylate. No excess liquid was
observed.
[0337] The filament was placed in a 1.7 ml microcentrifuge tube
with 1 ml 1.times.PBS. At daily intervals for 6 days, 150 .mu.L of
PBS was removed from each well and placed into a 96 well plate for
subsequent analysis. The remaining 850 .mu.L was removed from the
sample, and to the tube was added 1 ml of 1.times.PBS. After 6
days, the filament was placed in a 1.7 ml microcentrifuge tube with
1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL. At daily
intervals for 35 days, 150 .mu.L of PBS was removed from each well
and placed into a 96 well plate for subsequent analysis. The
remaining 850 .mu.L was removed from the sample, and to the tube
was added 1 ml of fresh 1.times.PBS containing alpha-Amylase at
0.121 .mu.g/mL. The 96-well plate was analyzed for BSA using the
Quanitpro Assay Kit (Sigma). For the first 6 days, there was an
initial burst of BSA, followed by a very slow release. After the
addition of PBS+Amylase, the rate of BSA release significantly
increased, and was relatively constant over the next 35 days.
Results are shown in Table 2 and FIG. 1.
TABLE-US-00002 TABLE 2 Cumulative BSA release Timepoint (% of Total
BSA) 1 4.8 2 5.35 3 5.7 4 5.98 5 6.19 6 6.36 7 9.46 8 10.7 9 11.82
10 12.94 11 14.01 12 15.06 13 16.11 14 17.23 15 18.11 16 19.04 17
19.92 18 21.26 19 22.15 20 23.04 21 24.06 22 25.35 23 26.31 24
26.91 25 27.51 26 28.63 27 29.19 28 29.75 29 30.44 30 31.11 31
31.43 32 31.63 33 31.83 34 32.07 35 32.31 36 32.72 37 32.95 38
33.27 39 33.83 40 34.15 41 34.43 42 34.71
Example 26
Polyalditol-acrylate Filaments with Bioactive Agent and Release
[0338] Polyalditol-acrylate in an amount of 1,500 mg of as prepared
in Example 21 was placed in an 8 ml amber vial. To the PA-Acrylate
was added 1 mg of DBDS (lyophilized), 15 mg Bovine Serum Albumin,
and 1 mL of 1.times. phosphate-buffered saline (1.times.PBS). The
reagents were then mixed for one hour on a shaker at 37.degree. C.
The mixture in an amount of 10 uL was injected, using a 23 gauge
needle, into a 22 mm length opaque silicone tube (P/N 10-447-01;
Helix Medical, Carpinteria, Calif.). The tubing was placed into a
Dymax Lightweld PC-2 illumination system (Dymax Corp.; light
intensity 6.5 mW/cm.sup.2), 15 cm from light source, illuminated
for 270 seconds, and then removed. After illumination, the filament
was removed from the silicone tubing by rolling a pencil over the
tubing, starting from the back. The filament was firm, which
indicated complete polymerization of the polyalditol-acrylate. No
excess liquid was observed. The filament was manipulated with
forceps.
[0339] The filament was placed in a 1.7 ml microcentrifuge tube
with 1 ml PBS containing alpha-Amylase at 0.121 .mu.g/mL. At daily
intervals for 15 days, 150 .mu.l of PBS was removed from each well
and placed into a 96 well plate for subsequent analysis. The
remaining 850 .mu.L was removed from the sample, and to the tube
was added 1 ml of fresh PBS containing alpha-Amylase at 0.121
.mu.g/mL. The 96-well plate was analyzed for BSA using the
Quanitpro Assay Kit (Sigma).
Example 27
Maltodextrin-acrylate Filaments with Bioactive Agent and
Release
[0340] Maltodextrin filaments were synthesized using a 1,100 mg/mL
solution as described in example 25 using an anti-horseradish
peroxidase antibody (P7899; Sigma) instead of BSA. The filament
contained 800 ug of the anti-horseradish peroxidase antibody. The
filament was placed in a 1.7 ml microcentrifuge tube containing 1
ml of 1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL. At
daily intervals for 5 days, 100 .mu.l of PBS was removed from the
sample, placed into a 96 well plate and incubated for 60 minutes at
37.degree. C. The remaining 850 .mu.L was removed from the sample,
and replaced with 1 ml fresh 1.times.PBS containing alpha-Amylase
at 0.121 .mu.g/mL. After 1 hour, the plate was washed three times
with 1 ml PBS/Tween (Sigma). 150 ul StabilCoat.TM. Immunoassay
Stabilizer (SurModics, Eden Prairie, Minn.) was added to the well
and incubated for 30 minutes at room temperature. After 30 minutes,
the 96-well plate was washed three times with PBS/Tween. A solution
of 0.5 mg/ml Horseradish Peroxidase (Sigma) in 1.times.PBS (100 uL)
was added to the well and incubated for 60 minutes. After 60
minutes, the 96-well plate was washed six times with PBS/Tween. A
chromogenic assay was then performed. After 15 minutes, the 96 well
plate was analyzed for HU conjugate on a spectrophotometer (Tecan)
at 560 nm absorbance. Detectable Antibody was found at each time
point.
Example 28
Bioactive Agent Incorporation and Release from a
MD-Acrylate/photo-PVP-coated PEBAX Rod
[0341] 100 mg photo-PVP and 5 mg the photoinitiator tetra-BBE-PET
were mixed with 10 ml Isopropyl alcohol (IPA; Fisher) for 1 minute.
The mixture in an amount of 1 ml was placed into a 1.7 ml eppendorf
tube (VWR). A 1.2 cm PEBAX.RTM. rod (Medical Profiles, Inc) was
dipped into the solution for 10 seconds, at a dip rate of 0.75
cm/second, and then removed at the same rate. The rod was allowed
to air dry for 10 minutes. The rod was placed into a Dymax
Lightweld PC-2 illumination system (Dymax Corp.; light intensity
6.5 mW/cm.sup.2), 30 cm from light source, illuminated for 180
seconds, and then removed.
[0342] 1000 mg of MD-Acrylate, as prepared in Example 13 was placed
in an 8 mL amber vial. To the MD-Acrylate was added 5 mg DBDS, 1 ml
of 1.times. phosphate-buffered saline (PBS), and 100 mg BSA. The
reagents were then mixed for one hour on a shaker at 37.degree. C.
The mixture in an amount of 1 ml was placed into a 1.8 ml eppendorf
tube (VWR). The photo-PVP/tetra-BBE-PET coated PEBAX.RTM. rod was
dipped into the mixture for 30 seconds, at a dip rate of 0.3 cm/s,
and then removed at the same rate. The rod was immediately placed
into a Dymax Lightweld PC-2 illumination system (Dymax Corp.; light
intensity 6.5 mW/cm.sup.2), 30 cm from light source, and
illuminated for 180 seconds and then removed.
[0343] The rod was placed in a 1.7 ml microcentrifuge tube with 1
ml 1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL. At
intervals for 9 days, 150 .mu.l of PBS was removed from each well
and placed into a 96 well plate. The remaining 850 .mu.l were
removed from the samples, and replaced with 1 ml fresh 1.times.PBS
containing alpha-Amylase at 0.121 .mu.g/mL. The 96 well-plate was
analyzed for BSA release using the Quanitpro Assay Kit (Sigma). BSA
was detected at each timepoint. Results are shown in Table 3 and
FIG. 2.
TABLE-US-00003 TABLE 3 Cumulative BSA release Timepoint (% of Total
BSA) 1 7.0 2 15.0 3 19.1 4 22.7 5 25.6 7 28.6 8 31.5 9 34.2
Example 29
Degradation of MD-acrylate Coating
[0344] The MD-Acrylate-coated PEBAX rod (as prepared in Example 28)
was placed in 5 ml of 1.times. phosphate-buffered saline (PBS)
containing alpha-Amylase at 24 .mu.g/mL for 7 days on a rotating
plate at 37.degree. C. After 7 days, the rod was removed from the
PBS and washed with distilled water. The rod was then examined
under a Scanning Electron Microscope (LEO Supra 35 VP); upon
examination, no trace of the MD-Acrylate coating was detected.
Example 30
Degradation of MD-Acrylate Filament in Vitreal Fluid
[0345] A circumferential dissection of the anterior segment
(cornea, aqueous humour, lens) of porcine eye was performed, and
the vitreous was squeezed out from the globe into a 20 mL amber
vial; approx 10 mL total was retrieved from a total of four eyes.
200 mg/mL and 1100 mg/mL Maltodextrin filaments, formed in example
21, were placed into 2 mL of the vitreous solution, and placed at
37.degree. C. on a rotator plate. The 200 mg/mL filament had
completely dissolved after 24 hours. The 1,100 mg/mL filaments
completely degraded after 30 days in the vitreous.
Example 31
Formation of a Maltodextrin-acrylate Biodegradable Matrix Using
REDOX Chemistry
[0346] Two solutions were prepared. Solution #1 was prepared as
follows: 250 mg of MD-acrylate as prepared in example 13 was placed
in an 8 mL vial. To the MD-acrylate was added 15 mg ferrous
gluconate hydrate (Sigma), 30 mg Ascorbic Acid (Sigma), 67 uL AMPS
(Lubrizol) and 1,000 uL deionized water. Solution #2 was prepared
as follows: 250 mg of MD-acrylate as prepared in example 13 was
placed in a second 8 mL vial. To this MD-acrylate was added 30 uL
AMPS, 80 uL Hydrogen Peroxide (Sigma) and 890 uL 0.1 M Acetate
buffer (pH 5.5).
[0347] 50 uL of Solution #1 was added to a glass slide. 50 uL of
solution #2 was added to Solution #1 with slight vortexing. After
mixing for 2 seconds, the mixture polymerized and formed a
semi-firm gel having elastomeric properties.
Example 32
Formation of Maltodextrin-acrylate Biodegradable Matrix Using REDOX
Chemistry
[0348] Two solutions were prepared, similar to Example 31, but in
this Example Solution #1 different concentrations of ferrous
gluconate hydrate (Sigma) and ascorbic acid were used. Solution #1
was prepared as follows: 250 mg of MD-acrylate (as prepared in
example 13) was placed in an 8 mL vial. To the MD-acrylate was
added 5 mg ferrous gluconate hydrate (Sigma), 40 mg ascorbic acid
(Sigma), 67 uL AMPS (Lubrizol) and 1,000 uL deionized water.
Solution #2 was prepared as follows: 250 mg of MD-acrylate as
prepared in example 7 was placed in a second 8 mL vial. To this
MD-acrylate was added 30 uL AMPS, 80 uL Hydrogen Peroxide (Sigma)
and 890 uL 0.1 M Acetate buffer (pH 5.5).
[0349] 50 uL of Solution #1 was added to a glass slide. 50 uL of
solution #2 was added to Solution #1 with slight vortexing. After
mixing for 8 seconds, the mixture polymerized and formed a
semi-firm gel having elastomeric properties.
Example 33
Formation of Maltodextrin-acrylate Biodegradable Matrix using REDOX
Chemistry
[0350] Two solutions were prepared. Solution #1 was prepared as
follows: 250 mg of MD-acrylate (as prepared in example 13) was
placed in an 8 mL vial. To the MD-acrylate was added 15 mg Iron
(II) L-Ascorbate (Sigma), 30 mg Ascorbic Acid (Sigma), 67 uL AMPS
(Lubrizol) and 1,000 uL deionized water. Solution #2 was prepared
as follows: 250 mg of MD-acrylate as prepared in example 7 was
placed in a second 8 mL vial. To this MD-acrylate was added 30 uL
AMPS, 80 uL hydrogen peroxide (Sigma) and 890 uL 0.1 M Acetate
buffer (pH 5.5).
[0351] 50 uL of Solution #1 was added to a glass slide. 50 uL of
solution #2 was added to Solution #1 with slight vortexing. After
mixing for 2 seconds, the mixture polymerized and formed a
semi-firm gel having elastomeric properties.
Example 34
Formation of Polyalditol-acrylate Biodegradable Matrix Using REDOX
Chemistry
[0352] Two solutions were prepared. Solution #1 was prepared as
follows: 1,000 mg of Polyalditol-acrylate as prepared in Example 21
was placed in an 8 mL vial. To the Polyalditol-acrylate was added
15 mg Ferrous Sulfate Heptahydrate (Sigma), 30 mg Ascorbic Acid
(Sigma), 67 uL AMPS (Lubrizol) and 1,000 uL deionized water;
Solution #2 was prepared as follows: 1,000 mg of
Polyalditol-acrylate as prepared in example was placed in a second
8 mL vial. To this Polyalditol-acrylate was added 30 uL AMPS, 80 uL
Hydrogen Peroxide (Sigma) and 890 uL 0.1 M Acetate buffer (pH
5.5).
[0353] 50 uL of Solution #1 was added to a glass slide. 50 uL of
solution #2 was added to Solution #1 with slight vortexing. After
mixing for 2 seconds, the mixture polymerized and formed a
semi-firm gel having elastomeric properties.
Example 35
Coating a PEBAX.RTM. Rod with MD-Acrylate Using REDOX
[0354] Photo-PVP (100 mg) and the photoinitiator tetra-BBE-PET (5
mg), were mixed with 10 mL isopropyl alcohol (IPA; Fisher) for 1
minute. The mixture in an amount of 1 mL was placed into a 1.8 mL
eppendorf tube (VWR). A 1.2 cm PEBAX.RTM. rod (Medical Profiles,
Inc) was dipped into the solution for 10 seconds, at a dip rate of
0.1 cm/second, and then removed at the same rate. The rod was
allowed to air dry for 5 minutes. The rod was placed into a Dymax
Lightweld PC-2 illumination system (Dymax Corp.; light intensity
6.5 mW/cm.sup.2), 30 cm from light source, illuminated for 180
seconds, and then removed.
[0355] Two solutions were prepared. Solution #1 was prepared as
follows: 250 mg of MD-acrylate (as prepared in example 13) was
placed in an 8 ml vial. To the MD-acrylate was added 15 mg Iron
(II) Ascorbate (Sigma), 30 mg Ascorbic Acid (Sigma), 67 ul AMPS
(Lubrizol) and 1,000 ul deionized water. Solution #2 was prepared
as follows: 250 mg of MD-acrylate was placed in a second 8 ml vial.
To this MD-acrylate was added 30 ul AMPS, 80 ul Hydrogen Peroxide
(Sigma) and 890 ul 0.1 M Acetate buffer (pH 5.5).
[0356] Solution #1 in an amount of 1 mL was placed into a 1.8 mL
eppendorf tube (VWR). The photo-PVP/tetra-BBE-PET coated PEBAX.RTM.
rod was dipped into the mixture for 30 seconds, at a dip rate of
0.5 cm/s, and then removed at the same rate. The PEBAX rod was
allowed to air dry for 10 minutes. Solution #2 in an amount of 1 mL
was placed into a second 1.8 mL eppendorf tube (VWR). The
photo-PVP/tetra-BBE-PET and Solution #1 coated PEBAX.RTM. rod was
dipped into the mixture for 30 seconds, at a dip rate of 0.5 cm/s,
and then removed at the same rate.
[0357] The MD-Acrylate coated rod was examined under Scanning
Electron Microscope (SEM; LEO Supra 35 VP); the MD-Acrylate coating
thickness ranged from 15 to 20 um, with an average thickness of
16.8 um.
Example 36
Bioactive Agent Incorporation into a MD-Acrylate Matrix
[0358] Two solutions were prepared. Solution #1 was prepared as
follows: 250 mg of MD-acrylate (as prepared in example 13) was
placed in an 8 ml vial. To the MD-acrylate was added 15 mg Iron
(II) Acetate (Sigma), 30 mg Ascorbic Acid (Sigma), 67 ul AMPS
(Lubrizol), 75 mg Bovine Serum Albumin (BSA; representing the
bioactive agent) and 1,000 .mu.L deionized water. Solution #1 was
prepared as follows: 250 mg of MD-acrylate was placed in a second 8
ml vial. To this MD-acrylate was added 30 .mu.L AMPS, 80 .mu.L
Hydrogen Peroxide (Sigma), 75 mg BSA and 890 .mu.L Acetate buffer
(pH 5.5).
[0359] 50 .mu.L of Solution #1 was added to a glass slide. 50 .mu.L
of solution #2 was added to Solution #1 with slight vortexing.
After mixing for 2 seconds, the mixture polymerized and formed a
semi-firm gel having elastomeric properties.
Example 37
Enzyme Degradation of a MD-Acrylate Matrix Formed by REDOX
[0360] Maltodextrin-acrylate filaments were prepared using the
reagents at concentrations as described in Example 31. These
filaments were placed in microcentrifuge tubes containing 1 ml
either Phosphate Buffered Saline (PBS) or 1.times.PBS containing
alpha-Amylase at 0.121 .mu.g/mL. The tubes were then placed in an
incubator at 37.degree. C.
[0361] After 4 days in the 1.times.PBS containing alpha-Amylase at
0.121 .mu.g/mL, the 250 mg/mL filament had completely degraded,
leaving no trace of the matrix. The matrix in PBS showed no signs
of degradation.
Example 38
FAB Fragment Incorporation and Release from a MD-Acrylate
Filament
[0362] 600 milligrams of MD-Acrylate as prepared in Example 13 was
placed in an 8 mL amber vial. To the MD-Acrylate was added 5 mg of
DBDS (lyophilized), 10 mg Rabbit Anti-Goat Fragment Antibody
(catalog #300-007-003; Jackson Immunological Research, West Grove,
Pa.) and 1 mL of 1.times. phosphate-buffered saline (PBS). The
reagents were then mixed for one hour on a shaker at 37.degree. C.
The mixture in an amount of 10 .mu.L was pipetted into a 22 mm
length opaque silicone tube (P/N 10-447-01; Helix Medical,
Carpinteria, Calif.). The tubing was placed into a Dymax Lightweld
PC-2 illumination system (Dymax Corp.; light intensity 6.5
mW/cm.sup.2), 15 cm from light source, illuminated for 270 seconds,
and then removed. After illumination, the filament was removed from
the silicone tubing by rolling a pencil over the tubing, starting
from the back. The filament was firm and completely crosslinked,
with no excess liquid. The filament was placed in a 1.7 mL
microcentrifuge tube with 0.5 ml 1.times.PBS containing
alpha-Amylase at 0.121 .mu.g/mL (eluent solution). At predetermined
intervals for 17 days, 200 .mu.L of the eluent solution was removed
from each tube, and 100 .mu.L was placed into two 96 well plates.
The remaining 300 pit were removed from the samples, and replaced
with 0.5 mL fresh 1.times.PBS containing alpha-Amylase at 0.121
.mu.g/mL. The 96 well plates were analyzed for total FAB molecule
release and FAB activity using an Enzyme-Linked Immunosorbent Assay
(ELISA). Briefly, the 100 .mu.L eluent solution was incubated at
37.degree. C. for one hour and then washed 3.times. with 2 ml
PBS/Tween 20 (Sigma). The wells were blocked with 100 .mu.L
StabilCoat.TM. for 1 hour at room temperature and then washed
3.times. with 2 mL PBS/Tween 20. 100 uL of either 0.1 ug/mL (in
PBS/Tween) HRP-labeled Goat IgG (Jackson Immunological; catalog
#005-030-003) for molecule activity or 0.08 ug/mL (in PBS/Tween)
HRP-labeled Goat anti-Rabbit IgG (Jackson Immunological; catalog
#111-305-003) was incubated for 1 hour at 37.degree. C. The wells
were washed 6.times. with 2 mL PBS/Tween 20. 100 .mu.L of TMB
Microwell Peroxidase Substrate System (KPL, Catalog #50-76-00;
Gaithersburg, Md.) as added to each well. After 15 minutes, the 96
well plate was analyzed for HRP conjugate on a spectrophotometer
(Tecan) at 650 nm absorbance. Detectable Antibody was found at each
timepoint. Results are shown in Table 4 and FIG. 3.
TABLE-US-00004 TABLE 4 Fab Fragment release ABS values Cumulative
Active FAB Cumulative Total Fab Timepoint (Day) Abs at 650 nm Abs
at 650 nm 1 1.37 1.97 3 3.12 4.07 4 4.54 5.87 6 5.69 7.54 7 6.12
8.60 8 6.53 9.01 10 6.94 9.79 13 7.34 10.64 15 7.54 11.18 17 7.71
11.62 19 7.81 11.92 21 7.90 12.28 23 8.00 12.68 26 8.09 13.11
Example 39
Rabbit Antibody Incorporation and Release from a MD-acrylate (Redox
Polymerization) Coated Stainless Steel Rod
[0363] 316V or 304V stainless steel rods (0.035'' diameter; Small
Parts, Inc.) were cleaned by wiping the rods with isopropyl alcohol
(IPA) soaked Alpha 10 clean-wipes (Texwipe; Kernersville, N.C.) and
then sonicating the rods in a solution of 10% Valtron SP2200
detergent (Valtech Corp.) in hot tap water for 10 minutes. The rods
were then rinsed 3.times. with deionized water followed by a 1
minute sonication in hot tap water; the rinse and sonication steps
were then repeated.
[0364] A 0.5% 1,4-bis(trimethoxysilylethyl)benzene (B2495.6, UCT,
Bristol, Pa.) solution in 89.5% IPA and 10% deionized water was
prepared (See Example 1 of U.S. Pat. No. 6,706,408B2-) and the
cleaned rods were dipped into the silane solution within 2 minutes
of coming out of the deionized rinse. The rods were allowed to
dwell in the silane solution for 3 minutes and then pulled out at a
rate of 1.0 cm/s. They were allowed to air dry for 2 minutes at
room temperature and then placed at 110.degree. C. for 5
minutes.
[0365] A solution of photo-PVP (15 mg/mL), DBDS (1 mg/mL),
tetra-BBE-PET 0.075 (mg/mL) and PVP K90 (20 mg/ml; BASF) was
prepared in 60% IPA/40% water) (see also Example 1 of U.S. Pat. No.
6,706,408B2). The silane treated rods were dip coated into the
above solution using the following parameters:
[0366] Velocity in =2.0 cm/s; dwell time=60 seconds; velocity
out=0.1 cm/s The coated rods were allowed to air dry at room
temperature for 5 minutes and then illuminated in a Dymax lamp UV
light chamber, with rotation, for 3 minutes. The dipping procedure
was then repeated with a dwell time of 120 seconds (all other
dipping parameters the same).
[0367] Two solutions were prepared. The first solution (#1) was
prepared by placing 600 mg of MD-acrylate (as prepared in example
13) into an 8 mL vial and then adding 9 mg iron (II) ascorbate
(Sigma), 30 mg ascorbic acid (Sigma), 67 .mu.L AMPS (Lubrizol), 16
mg Rabbit Anti-HRP antibody (Sigma; catalog #P7899), and 1,000
.mu.L deionized water. The second solution (#2) was prepared by
placing 600 mg of MD-acrylate in a second 8 mL vial and then adding
30 .mu.L AMPS, 16 mg Rabbit Anti-HRP antibody (Sigma; catalog
#P7899), 80 .mu.l, hydrogen peroxide (Sigma) and 890 .mu.L 0.1 M
Acetate buffer (pH 5.5).
[0368] Solution #1 in an amount of 1 mL was placed into a 1.8 mL
eppendorf tube (VWR). The (PV01/K90/DBDS/tetra-BBE-PET)-coated
stainless steel rod (20 mm in length) was dipped into the mixture
for 20 seconds, at a dip rate of 0.75 cm/s, and then removed at the
same rate. The rod was allowed to air dry for 10 minutes. Solution
#2 in an amount of 1 mL was placed into a second 1.8 mL eppendorf
tube (VWR). The (PV01/K90/DBDS/tetra-BBE-PET)-coated and Solution
#1 coated rod was dipped into the Solution #2 for 20 seconds, at a
dip rate of 0.75 cm/s, and then removed at the same rate. Contact
with Solution #2 initiated the redox reaction and caused formation
of an MD-Acrylate coated layer on the PV01/K90/DBDS/tetra-BBE-PET
coated layer; the MD-Acrylate coating was cured within 10
seconds
[0369] The coated rod was examined under Optical Inferometer
Microscope (Veeco) which revealed that the MD-Acrylate coating
layer had a thickness of 70
[0370] The MD-Acrylate coated rod was placed in a 0.6 mL
microcentrifuge tube with 0.5 ml 1.times.PBS containing
alpha-Amylase at 0.121 .mu.g/mL (eluent solution) to assess release
of the bioactive agent (antibody) from the coating.
[0371] At predetermined intervals for 19 days, 200 .mu.L of the
eluent solution was removed from the tube, which was divided into
two 100 .mu.L aliquots and placed into two 96 well plates. The
remaining 3004 was removed from the microcentrifuge tube, and 0.5
mL of fresh eluent solution (1.times.PBS containing alpha-Amylase
at 0.121 .mu.g/mL) was added to the microcentrifuge tube having the
MD-Acrylate coated rod. The eluent samples in 96 well plates were
analyzed for total Rabbit Antibody molecule release and activity
using an Enzyme-Linked Immunosorbent Assay (ELISA). Briefly, the
100 .mu.L of eluent solution was incubated at 37.degree. C. for one
hour and then washed 3.times. with 2 ml PBS/Tween 20 (Sigma). The
wells were blocked with 100 .mu.L StabilCoat.TM. (SurModics, Eden
Prairie, Minn.) for 1 hour at room temperature and then washed
3.times. with 2 ml PBS/Tween 20. 100 .mu.L of either 0.1 ug/mL (in
PBS/Tween) HRP (Sigma; catalog #P8375) for molecule activity or
0.08 ug/mL (in PBS/Tween) HRP-labeled Goat anti-Rabbit IgG (Jackson
Immunological; catalog #111-305-003) was incubated for 1 hour at
37.degree. C. The wells were washed 6.times. with 2 ml PBS/TWeen
20. 100 .mu.L of TMB Microwell Peroxidase Substrate System (KPL,
Catalog #50-76-00; Gaithersburg, Md.) was added to each well. After
15 minutes, the 96 well plate was analyzed for HRP conjugate on a
spectrophotometer (Tecan) at 650 nm absorbance. Detectable antibody
was found in the eluate samples at each timepoint. Results are
shown in Table 5 and FIG. 4.
TABLE-US-00005 TABLE 5 Cumulative Active IgG Cumulative Total IgG
Timepoint (Day) Abs at 650 nm Abs at 650 nm 1 1.08 2.01 2 1.82 3.78
4 2.01 5.11 6 2.41 5.57 8 2.48 5.75 10 2.54 5.88 12 2.59 5.97 19
2.87 7.88
Example 40
Rabbit Antibody Incorporation and Release from a MD-acrylate
(Photo-Initiated Polymerization) Coated Stainless Steel rod
[0372] 316V or 304V stainless steel rods (0.035'' diameter; Small
Parts, Inc., Miami Lakes, Fla.) were cleaned by wiping the rods
with isopropyl alcohol (IPA)-soaked Alpha 10 clean-wipe (Texwipe)
and then sonicating the rods in a solution of 10% Valtron SP2200
detergent in hot tap water for 10 minutes. The rods were then
rinsed 3.times. with deionized water followed by a 1 minute
sonication in hot tap water; the rinse and sonication step was then
repeated.
[0373] The cleaned rods were dipped into a 0.5%
1,4-bis(trimethoxysilylethyl) benzene solution (as described in
Example 39) within 2 minutes of coming out of the deionized rinse.
The rods were allowed to dwell in the silane solution for 3 minutes
and then pulled out at a rate of 1.0 cm/s. The rods were allowed to
air dry for 2 minutes at room temperature and then placed at
110.degree. C. for 5 minutes.
[0374] A solution of PV01 (15 mg/mL), DBDS (1 mg/mL), tetra-BBE-PET
(0.075 mg/ml) and K90 (20 mg/mL; BASF) was prepared in 60% IPA/40%
water. The silane treated rods were dip coated into the above
solution using the following parameters:
[0375] Velocity in =2.0 cm/s; dwell time=60 seconds; velocity
out=0.1 cm/s. The coated rods were allowed to air dry at room
temperature for 5 minutes and then illuminated in a Dymax lamp UV
light chamber, with rotation, for 3 minutes. The dipping procedure
was then repeated with a dwell time of 120 seconds (all other
dipping parameters the same).
[0376] 600 milligrams of MD-Acrylate as prepared in Example 13 was
placed in an 8 mL amber vial. To the MD-Acrylate was added 5 mg of
DBDS (lyophilized), 16 mg Rabbit Anti-HRP antibody (Sigma; catalog
#P7899) and 1 ml of 1.times. phosphate-buffered saline (PBS). The
reagents were then mixed for one hour on a shaker at 37.degree. C.
The MD-Acrylate solution in an amount of 1 mL was placed into a 1.8
mL eppendorf tube (VWR). The (PV01/K90/DBDS/tetra-BBE-PET)-coated
stainless steel rod (20 mm in length) was dipped into the mixture
for 20 seconds, at a dip rate of 0.75 cm/s, and then removed at the
same rate. The rod was then immediately illuminated in a Dymax lamp
UV light chamber, with rotation, for 3 minutes. The rod was allowed
to air dry for 5 minutes, and the dipping and illuminating
procedures were repeated one more time.
[0377] The MD-Acrylate coated rod was examined under Optical
Inferometer Microscope (Veeco); the MD-Acrylate coating layer had a
coating thickness of 20 .mu.m.
[0378] The MD-Acrylate coated rod was placed in a 0.6 mL
microcentrifuge tube with 0.5 mL 1.times.PBS containing
alpha-Amylase at 0.121 .mu.g/mL (eluent solution) to assess
degradation of the coating and release of the bioactive agent
(antibody) from the coating. At predetermined intervals for 25
days, 200 .mu.L of the eluent solution was removed from each tube,
which was divided into two 100 .mu.L aliquots and placed into two
96 well plates. The remaining 300 .mu.L, was removed from the
microcentrifuge tube, and 0.5 mL of fresh eluent solution
(1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL) was added
to the microcentrifuge tube having the MD-Acrylate coated rod.
[0379] The 96 well plates were analyzed for total Rabbit Antibody
molecule release and activity using an Enzyme-Linked Immunosorbent
Assay (ELISA). Briefly, the 100 .mu.L eluent solution was added to
the wells and incubated at 37.degree. C. for one hour and then
washed 3.times. with 2 mL PBS/Tween 20 (Sigma). The wells were
blocked with 100 .mu.L StabilCoat.TM. for 1 hour at room
temperature and then washed 3.times. with 2 mL PBS/Tween 20. 100
.mu.L of either 0.1 ug/mL (in PBS/Tween) HRP (Sigma; catalog
#P8375) for molecule activity or 0.08 ug/mL (in PBS/Tween)
HRP-labeled Goat anti-Rabbit IgG (Jackson Immunological; catalog
#111-305-003) was incubated for 1 hour at 37.degree. C. The wells
were washed 6.times. with 2 mL PBS/Tween 20. 100 .mu.L of TMB
Microwell Peroxidase Substrate System (KPL, Catalog #50-76-00;
Gaithersburg, Md.) was added to each well. After 15 minutes, the 96
well plate was analyzed for HRP conjugate on a spectrophotometer
(Tecan) at 650 nm absorbance. Detectable antibody was found in the
eluate samples at each time point.
[0380] Based on the initial weight of the coating before
degradation, a theoretical maximum concentration of the antibody
was calculated. In addition to measurements performed by ELISA, the
MD-Acrylate coated rod was weighed at various time points to
determine the amount of material of the MD-Acrylate coated layer
lost from the rods due to amylase digestion. Results are shown in
Table 6 and FIG. 5.
TABLE-US-00006 TABLE 6 Cumulative Cumulative Maximum Active IgG
Total IgG MD-acrylate theoretical Timepoint release (%) release (%)
coating total IgG (Day) (ELISA) (ELISA) remaining (%) release (%) 1
7.14 9.29 2 8.14 10.14 83 17 4 8.49 10.29 6 8.93 10.49 7 80 20 8
9.27 10.76 10 9.63 10.90 12 10.06 11.19 14 10.19 11.36 80 20 17
10.65 11.91 19 11.19 12.68 22 12.62 13.38 25 14.76 14.35
Example 41
Rabbit Antibody Incorporation and Release from a MD-Acrylate
Filament
[0381] 600 milligrams of MD-Acrylate as prepared in Example 13 was
placed in an 8 ml amber vial. To the MD-Acrylate was added 5 mg of
DBDS (lyophilized), 16 mg Rabbit Antibody Anti-HRP (Sigma; catalog
#P7899) and 1 ml of 1.times. phosphate-buffered saline (PBS). The
reagents were then mixed for one hour on a shaker at 37.degree. C.
The mixture in an amount of 10 .mu.L was pipetted into a 22 mm
length opaque silicone tube (P/N 10-447-01; Helix Medical,
Carpinteria, Calif.). The tubing was placed into a Dymax Lightweld
PC-2 illumination system (Dymax Corp.; light intensity 6.5
mW/cm.sup.2), 15 cm from light source, illuminated for 270 seconds,
and then removed. After illumination, the filament was removed from
the silicone tubing by rolling a pencil over the tubing, starting
from the back. The filament was firm and completely crosslinked,
with no excess liquid.
[0382] The filament was placed in a 1.7 ml microcentrifuge tube
with 0.5 ml 1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL
(eluent solution). At predetermined intervals for 25 days, 200
.mu.l of the eluent solution was removed from each tube, and 100
.mu.L was placed into two 96 well plates. The remaining 300 .mu.l
were removed from the samples, and replaced with 0.5 ml fresh
1.times.PBS containing alpha-Amylase at 0.121 .mu.g/mL. The 96
wellplates were analyzed for total Rabbit Antibody molecule release
and activity using an Enzyme-Linked Immunosorbent Assay (ELISA).
Briefly, the 100 .mu.L eluent solution was added to the wells and
incubated at 37 degrees C. for one hour and then washed 3.times.
with 2 ml PBS/Tween 20 (Sigma). The wells were blocked with 100
.mu.L StabilCoat.TM. (SurModics) for 1 hour at room temperature and
then washed 3.times. with 2 ml PBS/Tween 20. 100 .mu.L of either
0.1 ug/ml (in PBS/Tween) HRP (Sigma; catalog #P8375) for molecule
activity or 0.08 ug/ml (in PBS/Tween) HRP-labeled Goat anti-Rabbit
IgG (Jackson Immunological; catalog #111-305-003) was incubated for
1 hour at 37 degrees C. The wells were washed 6.times. with 2 ml
PBS/Tween 20. 100 .mu.L of TMB Microwell
[0383] Peroxidase Substrate System (KPL, Catalog #50-76-00;
Gaithersburg, Md.) was added to each well. After 15 minutes, the 96
well plate was analyzed for HRP conjugate on a spectrophotometer
(Tecan) at 650 nm absorbance. Detectable Antibody was found at each
time point.
[0384] Results are shown in Table 7 and FIG. 6.
TABLE-US-00007 TABLE 7 Cumulative Cumulative Maximum Active IgG
Total IgG MD-acrylate theoretical Timepoint release (%) release (%)
coating total IgG (Day) (ELISA) (ELISA) remaining (%) release (%) 1
5.56 5.31 2 12.13 11.94 4 18.38 19.13 6 27.75 22.88 7 83 17 8 33.50
25.44 10 37.63 27.44 12 39.50 28.31 14 40.75 28.57 59 31 17 41.75
28.76 19 42.75 28.98 21 40 60 22 43.44 29.67 25 44.31 30.67
Example 42
Mechanical Testing of MD-acrylate Discs Formed Via REDOX
Polymerization
[0385] MD-acrylate discs formed via redox polymerization of
MD-acrylate coating solutions were tested for mechanical
properties.
[0386] A first solution (#1) was prepared by placing 300 mg of
MD-acrylate as prepared in Example 13 into an 8 ml vial and then
adding 9 mg iron (II) ascorbate (Sigma), 30 mg ascorbic acid
(Sigma), 67 .mu.L AMPS (Lubrizol), and 1,000 .mu.L deionized water.
Solution #2 was prepared by placing 300 mg of MD-acrylate into a
second 8 ml vial and then adding 30 .mu.L AMPS, 80 .mu.L if
hydrogen peroxide (Sigma) and 890 .mu.L of 0.1 M Acetate buffer (pH
5.5).
[0387] Viscosity of the first and second solutions were determined
on a Brookfield Viscometer. The average viscosity for both
solutions was 10.9 cP.
[0388] The modulus of the formed matrix was determined by
rheological measurements. In order to perform rheological
measurements, the first and second solutions were combined on the
testing plate in the Rheometer (Rheometric Scientific; model
#SR-2000) and the mixture was allowed to polymerize to form a
matrix. Data recording began before sample was cured in plates.
Briefly, 100 .mu.L of solution #1 and 100 .mu.L of solution #2 were
mixed on the lower testing plate. As the matrix formed, the upper
testing plate was lowered to fully contact the mixture of the first
and second solutions as the mixture polymerized into a matrix. The
sample was cured within 15 seconds. This curing method ensured
maximum contact between the two testing plates resulting in more
accurate testing compared to pre-formed matrices being placed
between the testing plates. The resulting MD-acrylate matrix had
properties of an elastic solid with an elastic (storage) modulus
ranging from 27 kPa to 30 kPa, and a viscous (loss) modulus of only
about 1 kPa. Results are shown in Table 8 and FIG. 7.
TABLE-US-00008 TABLE 8 (Testing Conditions: Stress: 433 Pa; strain
1.6%; frequency: 1 radian/sec) G' G'' G* Time (Elastic (Storage
(Loss (seconds) Modulus; Pa) Modulus; Pa) Modulus; Pa) 247 26820.2
1300.5 26851.7 261 26908.5 1294.55 26939.6 274 26872 1299.28
26903.4 288 26943.8 1343.69 26977.3 301 27376.6 1380.43 27411.4 315
27327.7 1373.31 27362.2 329 27319.8 1376.27 27354.5 342 27274.8
1362.35 27308.8 356 27246.6 1369.38 27281 369 27180.6 1373.6
27215.3 383 27174.4 1371.61 27209 397 27119.4 1366.76 27153.9 410
27105.4 1360.49 27139.5 424 27064.1 1358.45 27098.2 437 27019.9
1355.9 27053.9 451 27019.8 1355.39 27053.8 465 26972.3 1355.85
27006.4 478 26956.2 1361.11 26990.5 492 26918.2 1352.58 26952.2 505
26880.7 1355.85 26914.9 519 26840.4 1360.47 26874.9
Example 43
Lubricity and Durability Testing of MD-acrylate Coated PEBAX
Rods
[0389] To assess lubricity and tenacity of MD-acrylate coated
parts, frictional force testing was performed.
[0390] Frictional testing over the last 40 cycles of a 50 cycle
test was evaluated. Coated rods were evaluated by a horizontal sled
style friction test method (modified ASTM D-1894, as described
below). Silicone Pads (7 mm diameter) were hydrated and then
wrapped around a 200 gram stainless steel sled. The silicone pad
was clipped together tightly on the opposite side of the sled. The
sled with rotatable arm was then attached to a 500 gram Chatillon
Digital Force Gauge (DGGHS, 500.times.0.1) with computer interface.
The testing surface was mounted on a 22.5 inch positioning rail
table with micro-stepper motor control (Compumotor SX6
Indexer/Drive).
[0391] MD-coated rods (from example 18) were hydrated in deionized
water and clamped onto the test surface 1 inch (or approximately
2.5 cm) apart. The hydrated Silicone pad (jaw force set at 500 g)
moved at 0.5 cm/sec over a 5 cm section for 50 push/pull cycles,
and the final force measurements were taken over the last 40
push/pull cycles.
[0392] As shown in Table 9 and FIG. 8, compared to the benchmark
synthetic coating (see example 1 of U.S. Pat. No. 6,706,408B2) the
MD-acrylate coated rod provided a coating having excellent
lubricity and durability; the rods coated with a MD-acrylate
formulation containing a photoinitiator, the grams of force
remained relatively constant for the last 40 cycles, indicating a
durable coating.
TABLE-US-00009 TABLE 9 MD-acrylate photo coated rod Synthetic 1
9.67 12.53 2 9.53 12.43 3 9.45 12.26 4 9.18 12.40 5 9.12 12.33 6
9.13 12.17 7 9.11 12.15 8 9.06 12.12 9 8.92 12.25 10 8.96 12.19 11
8.91 12.11 12 8.98 12.02 13 8.95 12.02 14 8.82 12.25 15 8.79 12.10
16 8.84 12.08 17 8.85 12.02 18 8.89 11.92 19 8.76 12.10 20 8.81
12.08 21 8.85 11.93 22 8.85 11.86 23 8.88 11.83 24 8.81 12.01 25
8.90 11.94 26 8.96 11.94 27 9.04 11.87 28 9.04 11.71 29 8.97 12.00
30 9.03 11.94 31 9.05 11.79 32 9.20 11.83 33 9.32 11.75 34 9.23
11.90 35 9.22 11.82 36 9.39 11.79 37 9.46 11.68 38 9.52 11.74 39
9.53 11.91 40 9.56 11.87
Example 44
Preparation of Acylated Maltodextrin (Butyrylated-MD)
[0393] Maltodextrin having pendent butyryl groups were prepared by
coupling butyric anhydride at varying molar ratios.
[0394] To provide butyrylated-MD (1 butyl/4 glucose units, 1:4
B/GU) the following procedure was performed. Maltodextrin (MD;
Aldrich; 11.0 g; 3.67 mmole; DE (Dextrose Equivalent): 4.0-7.0) was
dissolved in dimethylsulfoxide (DMSO) 600 mL with stirring. The
size of the maltodextrin was calculated to be in the range of 2,000
Da-4,000 Da. Once the reaction solution was complete,
1-methylimidazole (Aldrich; 2.0 g, 1.9 mls) and butyric anhydride
(Aldrich; 5.0 g, 5.2 mls) was added with stirring. The reaction
mixture was stirred for four hours at room temperature. After this
time, the reaction mixture was quenched with water and dialyzed
against DI water using 1,000 MWCO dialysis tubing. The butyrylated
starch was isolated via lyophylization to give 9.315 g (85% yield).
NMR confirmed a butyrylation of 1:3 B/GU (1.99 mmoles butyl/g
sample).
[0395] To provide butyrylated-MD (1:8 B/GU), 2.5 g (2.6 mL) butyric
anhydride was substituted for the amount of butyric anhydride
described above. A yield of 79% (8.741 g) was obtained. NMR
confirmed a butyrylation of 1:5 B/GU (1.31 mmoles butyl/g
sample).
[0396] To provide butyrylated-MD (1:2 B/GU), 10.0 g (10.4 mL)
butyric anhydride was substituted for the amount of butyric
anhydride described above. A yield of 96% (10.536 g) was obtained.
NMR confirmed a butyrylation of 1:2 B/GU (3.42 mmoles butyllg
sample).
Example 45
Preparation of Acrylated Acvlated Maltodextrin
(Butyrylated-MD-Acrylate)
[0397] Preparation of an Acylated Maltodextrin Macromer Having
Pendent Butyryl and acrylate groups prepared by coupling butyric
anhydride at varying molar ratios.
[0398] To provide butyrylated-MD-acrylate (1 butyl/4 glucose units,
1:4 B/GU) the following procedure was performed. MD-Acrylate
(Example 13; 1.1 g; 0.367 mmoles) was dissolved in
dimethylsulfoxide (DMSO) 60 mL with stirring. Once the reaction
solution was complete, 1-methylimidazole (0.20 g, 0.19 mls) and
butyric anhydride (0.50 g, 0.52 mls) was added with stirring. The
reaction mixture was stirred for four hours at room temperature.
After this time, the reaction mixture was quenched with water and
dialyzed against DI water using 1,000 MWCO dialysis tubing. The
butyrylated starch acrylate was isolated via lyophylization to give
821 mg (75% yield, material lost during isolation). NMR confirmed a
butyrylation of 1:3 B/GU (2.38 mmoles butyl/g sample).
Example 46
Preparation of Acrylated Acylated Maltodextrin
(Butyrylated-MD-Acrylate)
[0399] Maltodextrin having pendent butyryl and acrylate groups
prepared by coupling butyric anhydride at varying molar ratios.
[0400] To provide butyrylated-MD-acrylate the following procedure
is performed. Butyrylated-MD (Example 43; 1.0 g; 0.333 mmole) is
dissolved in dimethylsulfoxide (DMSO) 60 mL with stirring. Once the
reaction solution is complete, a solution of EA-NCO from Example 12
(353 mg; 2.50 mmole) is evaporated and dissolved in dried DMSO 1.0
mL). The two DMSO solutions are mixed and heated to 55.degree. C.
overnight. The DMSO solution is placed in dialysis tubing (1000
MWCO) and dialyzed against water for 3 days. The macromer solution
is filtered and lyophilized to give a white solid.
* * * * *