Title of Invention

COMPOSITIONS AND METHODS FOR REDUCING SCAR TISSUE FORMATION

Abstract A drug attached to a carrier, the drug being selected from the group consisting of sirolimus, tacrolimus, everolimus and the analogs and derivatives thereof, the carrier onto which the drug is attached being selected from the group consisting of microparticles, gels, xerogels, bioadhesives, foams and liquids.
Full Text

COMPOSITIONS AND METHODS
FOR REDUCING SCAR TISSUE FORMATION
Field of Invention
This invention is related to the field of tissue healing and excess scar prevention by
pharmacological activity. Specifically, this invention is related to the use of sirolimus,
tacrolimus and analogs of sirolimus (i.e., rapamycin and derivatives thereof) to reduce and/or
prevent post-surgical scar tissue formation and/or adhesions.
Background
Excess post-operative scar tissue formation, adhesions and blood vessel narrowing are
major problems following abdominal, neurological, spinal, vascular, thoracic or other types of
surgery using both classical open and arthroscopic/laparoscopic procedures.
Scar tissue forms as part of the natural healing process of an injury whereupon the
body usually initiates a full and swift wound healing response resulting in reconstructed,
repaired tissue. In certain instances, however, this normal healing process may result in
excessive scar tissue.
Following some kinds of surgery or injury, excess scar tissue production is a major
problem which influences the result of surgery and healing. In the eye, for example,
post-operative scarring can determine the outcome of surgery. This is particularly the case in
the blinding disease glaucoma, where several anti-scarring regimens are currently used to
improve glaucoma surgery results, but are of limited use clinically because of severe
complications. Other examples of excess scar tissue production negatively impacting the
outcome of surgery include adhesion lysis surgery, angioplasty, spinal surgery, vascular
surgery and heart surgery.
Previous attempts to solve problematic post-surgical scarring have used highly
cytotoxic mitosis inhibitors such as anthracycline, daunomycin, mitomycin C and doxorubin.
Kelleher, U.S. Patent No. 6,063,396. Similarly, intraluminal administration of cytostatic
agents are reported to inhibit gi reduce arterial restenosis. Kurtz et al, U.S. Patent No.
5,981,568.

The current state of the art is lacking in post-surgical and post-trauma treatments to
significantly reduce the formation of scar tissue using compounds having a low medical risk
and a high therapeutic benefit.
Definitions
The term "attached" as used herein, refers to any interaction between a medium or
carrier and a compound. Attachment may be reversible or irreversible. Such attachment may
be, but is not limited to, covalent bonding, ionic bonding, Van de Waal forces or friction, and
the like. A compound is attached to a medium or carrier if it is impregnated, incorporated,
coated, in suspension with, in solution with, mixed with, etc.
The term "contacting" as used herein, refers to any physical relationship between a
biological tissue and a pharmaceutical compound attached to a medium. Such physical
relationship may be, but is not limited to, spraying, layering, impregnation, interior placement
into or exterioT placement onto, and the like.
The term "wound" as used herein, denotes a bodily injury with disruption of the
normal integrity of tissue structures. In one sense, the term is intended to encompass a
"surgical site". In another sense, the term is intended to encompass wounds including, but not
limited to, contused wounds, incised wounds, lacerated wounds, non-penetrating wounds (i.e.,
i
wounds in which there is no disruption of the skin but there is injury to underlying
structures), open wounds, penetrating wound, perforating wounds, puncture wounds, septic
wounds, subcutaneous wounds, burn injuries etc. Conditions related to wounds or sores which
may be successfully treated according to the invention are skin diseases.
The term "surgical site" as used herein, refers to any opening in the skin or internal
organs performed for a specific medical purpose. The surgical site may be "open" where
medical personnel have direct physical access to the area of interest as in traditional surgery.
Alternatively, the surgical site may be "closed" where medical personnel perform procedures
using remote devices such as, but not limited to, catheters wherein fluoroscopes may be used
to visualize the activities and; endoscopes (i.e., Iaparoscopes) wherein fiber optic systems may
be used to visualize the activities. A surgical site may include, but is not limited to, organs,
muscles, tendons, ligaments, connective tissue and the like.

The term "organ" as used herein, include, without limitation, veins, arteries, lymphatic
vessels, esophagus, stomach, duodenum, jejunum, ileum, colon, rectum, urinary bladder,
ureters, gall bladder, bile ducts, pancreatic duct, pericardia! sac, peritoneum, and pleura.
The term "skin" is used herein, very broadly embraces the epidermal layer of the skin
and, if exposed, also the underlying dermal layer. Since the skin is the most exposed part of
the body, it is particularly susceptible to various kinds of injuries such as, but not limited to,
ruptures, cuts, abrasions, burns and frostbites or injuries arising from the various diseases.
The term "anastomosis" as used herein, refers to a surgical procedure where two
vessels or organs, each having a lumen, are placed in such proximity that growth is stimulated
and the two vessels or organs are joined by forming continuous tissue. Preferably, the bodily
organs to be joined are veins, arteries and portions of the intestinal tract. Most preferably, the
organs to be joined are arteries. One of skill in the art will recognize that an anastomosis
procedure contemplated by the present invention is amenable to use not only in all areas of
vascular surgery but also in other surgical procedures for joining organs. Examples of
anastomoses that can be performed include, but are not limited to, arterial anastomosis,
venous anastomosis, arterio-venous anastomosis, anastomosis of lymphatic vessels,
gastroesophageal anastomosis, gastroduodenal anastomosis, gastrojejunal anastomosis,
anastomosis between and among the jejunum, ileum, colon and rectum, ureterovesicular
anastomosis, anastomosis of the gall bladder or bile duct to the duodenum, and anastomosis of
the pancreatic duct to the duodenum. In addition, an anastomosis may join an artifical graft
to a bodily organ that has a lumen. In one embodiment, the present invention contemplates
contacting a medium with an arterio-venous anastomosis of a patient, wherein said patient
exhibits symptoms of end stage renal disease and is undergoing dialysis.
The term "communication" as used herein, refers to the ability of two organs to
exchange body fluids by flowing or diffusing from one organ to another in the manner
typically associated with the organ pair that has is been joined. Examples of fluids that might
flow through an anastomosis include, but are not limited to, liquid and semi-solids such as
blood, urine, lymphatic fluid, bile, pancreatic fluid, ingesta and purulent discharge.
The term "medium" as used herein, refers to any material, or combination of materials,
which serve as a carrier or vehicle for delivering of a compound to a treatment point (e.g.,

wound, surgical site etc.). For ail practical purposes, therefore, the term "medium" is
considered synonymous with the term "carrier". In one embodiment, a medium comprises a
carrier, wherein said carrier is attached to a drug or compound and said medium facilitates
delivery of said carrier to a treatment point. In another embodiment, a carrier comprises an
attached drug wherein said carrier facilitates delivery of said drug to a treatment point.
Preferably, a medium is selected from the group consisting of foams, gels (including, but not
limited to, hydrogels), xerogels, microparticles (i.e., microspheres, liposomes, microcapsules
etc.), bioadhesives and liquids. Specifically contemplated by the present invention is a
medium comprising combinations of microparticles with hydrogels, bioadhesives, foams or
liquids. Preferably, hydrogels, bioadhesives and foams comprise any one, or a combination
of, polymers contemplated herein. Any medium contemplated by this invention may comprise
a controlled release formulation. For example, in some cases a medium constitutes a drug
delivery system that provides a controlled and sustained release of drugs over a period of time
lasting approximately from 1 day to 6 months.
The term "xerogel" as used herein, refers to any device comprising a combination of
silicone and oxygen having a plurality of air bubbles and an entrapped compound. The
resultant glassy matrix is capable of a controlled release of an entrapped compound during the
dissolution of the matrix.
The term "material" as used herein refers to any chemical that is useful in the creation
of a medium. For example, a liposome medium is comprised of a phospholipid material; a
microparticle or hydrogel medium is comprised of a polymer material, wherein said polymer
material is exemplified by poly(lactide-co-glycolide) copolymers and hyaluronic acid.
The term "reduction in scar tissue formation" as used herein refers to any tissue
response that reflects an improvement in wound healing. Specifically, improvement in
conditions such as, but not limited to, hyperplasia or adverse reactions to post-cellular trauma
are contemplated. It is not contemplated that all scar tissue must be avoided. It is enough if
the amount of scarring or hyperplasia is reduced as compared to untreated patients.
The term "foam" as used herein, refers to a dispersion in which a large proportion of
gas, by volume, is in the form of gas bubbles and dispersed within a liquid, solid or gel. The
diameter of the bubbles are usually relatively larger than the thickness of the lamellae between

the bubbles.
The term "gel" as used herein, refers to any material forming, to various degrees, a
medium viscosity liquid or a jelly-like product when suspended in a solvent. A gel may also
encompass a solid or semisolid colloid containing a certain amount of water. These colloid
solutions are often referred to in the art as hydrosols. One specific type of gel is a hydrogel.
The term "hydrogel" as used herein, refers to any material forming, to various degrees, a
jelly-like product when suspended in a solvent, typically water or polar solvents comprising
such as, but not limited to, gelatin and pectin and fractions and derivatives thereof. Typically,
a hydrogel is capable of swelling in water and retains a significant portion of water within its
structure without dissolution. In one embodiment, the present invention contemplates a gel '
that is liquid at lower than body temperature and forms a firm gel when at body temperature.
The term "spray" as used herein, refers to any suspension of liquid or particles blown,
ejected into, or falling through the air. Sprays can be jets of fine particles or droplets. A
spray can be an aerosol.
An "aerosol" is herein defined as a suspension of liquid or solid particles of a
substance (or substances) in a gas, such as, but not limited to dispersions. Aerosols may
comprise solid or liquid dispersions. The present invention contemplates the generation of
aerosols by both atomizers and nebulizers of various types. An "atomizer" is an aerosol
generator without a baffle, whereas a "nebulizer" uses a baffle to produce smaller particles.
In one embodiment, the present invention contemplates using the commercially available
Aerogen™ aerosol generator which comprises a vibrational element and dome-shaped aperture
plate with tapered holes. When the plate vibrates several thousand times per second, a micro-
pumping action causes liquid to be drawn through the tapered holes, creating a low-velocity
aerosol with a precisely defined range of droplet sizes. The Aerogen™ aerosol generator does
not require propellant. "Baffling" is the interruption of forward motion by an object, i.e. by a
"baffle." Baffling can be achieved by having the aerosol hit the sides of the container or
tubing. More typically, a structure (such as a ball or, other barrier) is put in the path of the
aerosol (See e.g. U.S. Patent 5,642,730, hereby incorporated by reference). The present
invention contemplates the use of a baffle in order to slow the speed of the aerosol as it exits
the delivery device.

The tram "compound" or "drug" as used herein, refers to any pharmacologically active
substance capable of being administered which achieves a desired effect. Compounds or
drugs can be synthetic or organic, proteins or peptides, oligonucleotides or nucleotides,
polysaccharides or sugars. Compounds or drugs may have any of a variety of activities,
which may be stimulatory or inhibitory, such as antibiotic activity, antiviral activity,
antifungal activity, steroidal activity, cytotoxic, cytostatic, anti-proliferative, anti-
inflammatory, analgesic or anesthetic activity, or can be useful as contrast or other diagnostic
agents. In a preferred embodiment, the present invention contemplates compounds or drugs
that are capable of binding to the mTOR protein and either reduce wound and post-surgical
adhesions and/or reduce wound and post-surgical scarring. In another embodiment, the
present invention contemplates compounds or drugs that are cytostatic and are believed to
primarily act by interrupting the cell division cycle in the GO or Gl stage, thus inhibiting
proliferation without killing the cell. It is not intended that the term compound or drug refers
to any non-pharmaceutically active material such as, but not limited to, polymers or resins
intended for the creation of any one specific medium.
The term "rapamycin" as used herein refers to a compound represented by the drug
sirolimus. Rapamycin is an antifungal antibiotic which may be naturally extracted from a
streptomycetes, e.g., Streptomyces hygroscopicus, chemically synthesized or produced by
genetic engineering cell culture techniques.
The term "analog" as used herein, refers to any coumpound having substantial
structure-activity relationships to a parent compound such that the analog has similar
biochemical activity as the parent compound. For example, sirolimus has many analogs that
are substituted at either the 2-, 7- or 32- positions. One of skill in the art should understand
that the term "derivative" is used herein interchangeably with term "analog".
The term "administered" or "administering" a compound or drug, as used herein, refers
to any method of providing a compound or drug to a patient such that the compound or drug
has its intended effect on the patient. For example, one method of administering is by an
indirect mechanism using a medical device such as, but not limited to a catheter, spray gun,
syringe etc. A second exemplary method of administering is by a direct mechanism such as,
oral ingestion, transdermal patch, topical, inhalation, suppository etc.

The term "biocompatible", as used herein, refers to any material does not elicit a
substantial detrimental response in the host. There is always concern, when a foreign object
is introduced into a living body, that the object will induce an immune reaction, such as an
inflammatory response that will have negative effects on the host. In the context of this
invention, biocompatiblity is evaluated according to the application for which it was designed:
for example; a bandage is regarded a biocompable with the skin, whereas an implanted
medical device is regarded as biocompatible with the internal tissues of the body. Preferably,
biocompatible materials include, but are not limited to, biodegradable and biostable materials.
The term "biodegradable" as used herein, refers to any material that can be acted upon
biochemically by living cells or organisms, or processes thereof, including water, and broken
down into lower molecular weight products such that the molecular structure has been altered.
The term "bioerodible" as used herein, refers to any material that is mechanically worn
away from a surface to which it is attached without generating any long term inflammatory
effects such that the molecular structure has not been altered. In one sense, bioerosin
represents the final stages of "biodegradation" wherein stable low molecular weight products
undergo a final dissolution.
The term "bioresorbable" as used herein, refers to any material that is assimilated into
or across bodily tissues. The bioresorption process may utilize both biodegradation and/or
bioerosin.
The term "biostable" as used herein, refers to any material that remains within a
physiological environment for an intended duration resulting in a medically beneficial effect.
The term "supplemental pharmaceutical compound" as used herein, refers to any
medically safe compound administered as part of a medium as contemplated by this invention.
Administration of a medium comprising a supplemental pharmaceutical compound includes,
but is not limited to, systemic, local, implantation or any other means. A supplemental
pharmaceutical compound may have activities similar to, or different from a compound
capable being cytostatic or of binding to the mTOR protein. Preferably, supplemental
pharmaceutical compounds include, but are not limited to, antiinflammatory drugs,
corticosteriods, antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
The term "complementary pharmaceutical compound" as used herein, refers to any

medically safe compound administered separately from a medium as contemplated by this
invention. Administration of a complementary pharmaceutical compound includes, but is not
limited to, oral ingestion, transdermal patch, topical, inhalation, suppository etc. Preferably,
complementary pharmaceutical compounds include, but are not limited to, sirolimus,
tacrolimus, analogs of sirolimus, antiinflammatory drugs, corticosteroids, antithrombotics,
antibiotics, antivirals, analgesics and anesthetics.
The term "colloidal system" or "colloid" as used herein, refers to a substance that
consists of particles dispersed throughout another substance which are too small for resolution
with an ordinary light microscope but are incapable of passing through a semipermeable
membrane. It is not necessary for all three dimensions to be within the colloidal system:
fibers may exhibit only two dimensions as a colloid, and thin films may have only a single
dimension as a colloid. It is not necessary for the units of a colloidal system to be discrete:
continuous network structures, the basic units of which are of colloidal dimensions also fall in
this class (e:g. porous solids, gels and foams). A fluid colloidal system may be composed of
two or more components and called a sol, e.g. a protein sol, a gold sol, an emulsion, a
surfactant solution above the critical micelle concentration, or an aerosol. In a suspension
solid particles are dispersed in a liquid; a colloidal suspension is one in which the size of the
particles lies in the colloidal range.
The term "dose metering element" as used herein, is an element that controls the
amount of compound administered. The element can, but need not, measure the amount of
compound as it is administered. In a preferred embodiment, the element is characterized
simply as a container of defined volume (e.g., a reservoir). In a preferred embodiment, the
defined volume is filled by the manufacturer or hospital professional (e.g., nurse, pharmacist,
doctor, etc.) and the entire volume is administered. In another embodiment, the reservoir is
configured as a transparent or semi-transparent cylinder with visible measurement indicia (e.g.
markings, numbers, etc.) and the filling is done to a desired point (e.g. less than the entire
capacity) using the indicia as a guide.
The term "fluid driving element" as used herein, is an element that moves fluid in a
direction along the device. In some embodiments, the fluid driving element comprises a

plunger driven by compressed gas, said compressed gas stored in a canister. In other
embodiments, it comprises a pump. In still other embodiments, it comprises a hand actuated
plunger (in the manner of a syringe).
The term "patient" as used herein, is a human or animal and need not be hospitalized.
For example, out-patients, persons in nursing homes are "patients."
The term "medical device", as used herein, refers broadly to any apparatus used in
relation to a medical procedure. Specifically, any apparatus that contacts a patient during a
medical procedure or therapy is contemplated herein as a medical device. Similarly, any
apparatus that administers a compound or drug to a patient during a medical procedure or
therapy is contemplated herein as a medical device. "Direct medical implants" include, but
are not limited to, urinary and intravascular catheters, dialysis shunts, wound drain tubes, skin
sutures, vascular grafts and implantable meshes, intraocular devices, implantable drug delivery
systems and heart valves, and the like. "Wound care devices" include, but are not limited to,
general wound dressings, non-adherent dressings, burn dressings, biological graft materials,
tape closures and dressings, and surgical drapes. "Surgical devices" include, but are not
limited to, endoscope systems (i.e., catheters, vascular catheters, surgical tools such as
scalpels, retractors, and the like) and temporary drug delivery devices such as drug ports,
injection needles etc.. to administer the medium. A medical device is "coated" when a
medium comprising a cytostatic or antiproliferative drug (i.e., for example, sirolimus or an
analog of sirolimus) becomes attached to the surface of the medical device. This attachment
may be permanent or temporary. When temporary, the attachment may result in a controlled
release of a cytostatic or antiproliferative drug.
The term "cytostatic" refers to any compound whose principal mechanism of
antiproliferative action interferes with the progress of the cell cycle in the GO or Gl phase. In
one embodiment, sirolimus, tacrolimus or analogs of sirolimus are cytostatic and interfere
with (i.e., stop) the cell cycle from progressing out of the Gl phase.
The term "endoscope" refers to any medical device that is capable of being inserted
into a living body and used for tasks including, but not limited to, observing surgical
procedures, performing surgical procedures, applying medium to a surgical site. An
endoscope is illustrated by instruments including, but not limited to, an arthroscope, a

laparoscope, hysteroscope, cytoscope, etc. It is not intended to limit the use of an endoscope
to hollow organs. It is specifically contemplated that endoscopes, such as an arthroscope or a
laparoscope is inserted through the skin and courses to a closed surgical site.
The term "liquid" as used herein, refers to a minimally viscous medium that is applied
to a surgical site by methods including, but not limited to, spraying, pouring, squeezing,
spattering, squirting, and the like.
The term "dispense as a liquid" as used herein, refers to spraying, pouring, squeezing,
spattering, squirting, aad the like.
The term "liquid administration" as used herein, refers to any method by which a
medium comprises an ability to flow or stream, either in response to gravity or by pressure-
induced force.
The term "liquid spray" as used herein, refers to a liquid administration comprising the
generation of finely dispersed droplets in response to pressure-induced force, wherein the
finely dispersed droplets settle onto a surgical site by gravity.
The term "pourable liquids" as used herein, refers to a liquid administration comprising
the flowing or streaming of a low viscosity liquid in response to gravity. The present
invention contemplates low viscosity liquids (at room temperature) ranging from between 1
and 15,000 centipoise, preferably between 1 and 500 centipoise {i.e., similar to saturated
glucose solution) and more preferably between 1 and 250 centipoise (i.e., similar to motor
oil).
The term "squeezable liquids" as used herein, refers to a liquid administration
comprising the flowing or streaming of a high viscosity liquid in response to a pressure-
induced force. The present invention contemplates high viscosity liquids (at room
temperature) ranging from between 5,000 and 100,000 centipoise, preferably between 25,000
and 50,000 centipoise (i.e., similar to mayonnaise), more preferably between 15,000 and
25,000 centipoise (i.e., similar to molten glass), and more preferably between 5,000 and
15,000 centipoise (i.e., similar to honey).
The term, "microparticle" as used herein, refers to any microscopic carrier to which a
compound or drug may be attached. Preferably, microparticles contemplated by this invention

are capable of formulations having controlled release properties.
The term "PLGA" as used herein, refers to mixtures of polymers or copolymers of
lactic acid and glycolic acid. As used herein, lactide polymers are chemically equivalent to
lactic acid polymer and glycolide polymers are chemically equivalent to glycolic acid
polymers. In one embodiment, PLGA contemplates an alternating mixture of lactide and
glycolide polymers, and is referred to as a poly(lactide-co-glycolide) polymer.
Summary
This invention is related to the field of tissue healing and scar prevention. In one
embodiment, pharmaceutical compounds are used to reduce and/or prevent scar tissue
formation. In another embodiment, sirolimus, tacrolimus and analogs of sirolimus (i.e.,
sirolimus and it's derivatives) are used to reduce and/or prevent post-surgical scar tissue
formation. In another embodiment, compounds capable of interrupting the cell cycle at the
GO or Gl stage are used to reduce and/or prevent excess scar tissue. In another embodiment,
compounds capable of binding to the mTOR protein are used to reduce and/or prevent scar
tissue formation.
One aspect of the present invention contemplates a drug attached to a carrier, the drug
being selected from the group consisting of sirolimus, tacrolimus, everolimus and the analogs
and derivatives of the drug, the carrier onto which the drug is attached being selected from
the group consisting of microparticles, gels, xerogels, bioadhesives, foams and liquids. In one
embodiment the carrier comprises a biocompatible material. In another embodiment, the
carrier comprises a biodegradable material. In one embodiment, the microparticles are
selected from the group consisting of microspheres, microencapsulating particles,
microcapsules and liposomes. In one embodiment, the microparticle comprises a polymer
selected from the group consisting of poly(lactide-co-glycolide), aliphatic polyesters including,
but not limited to, poly-glycolic acid and poly-lactic acid, hyaluronic acid, modified
polysacchrides, chitosan, cellulose, dextran, polyurethanes, polyacrylic acids, psuedo-
poly(amino acids), polyhydroxybutrate-related copolymers, polyanhydrides,
polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids. In one embodiment
the carrier comprises a material selected from the group consisting of gelatin, collagen,

cellulose esters, dextran sulfate, pentosan polysulfate, chitin, saccharides, albumin, fibrin
sealants, synthetic polyvinyl pyrrolidone, polyethylene oxide, polypropylene oxide, block
polymers of polyethylene oxide and polypropylene oxide, polyethylene glycol, acrylates,
acrylamides, methacrylates including, but not limited to, 2-hydroxyethyl methacrylate,
poly(ortho esters), cyanoacrylates, gelatin-resorcin-aldehyde type bioadhesives, polyacrylic
acid and copolymers and block copolymers thereof. In another embodiment, the carrier
comprises a polymer selected from the group consisting of poly(lactide-co-glycolide), aliphatic
polyesters including, but not limited to, poly-glycolic acid and poly-lactic acid, hyaluronic
acid, modified polysacchrides, chitosan, cellulose, dextran, polyurethanes, polyacrylic acids,
psuedo-poly(amino acids), polyhydroxybutrate-related copolymers, polyanhydrides,
polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids. In one
embodiment, the carrier releases said drug in a controlled release manner. In one
embodiment, the carrier is colored. In one embodiment, the carrier further comprises a radio-
opaque marker, wherein said marker is visualized by X-r4y spectroscopy.
One aspect of the present invention contemplates medium, comprising a compound
selected from the group consisting of sirolimus, tacrolimus, analogs of sirolimus and
pharmaceutically acceptable salts thereof, wherein said medium is selected from the group
consisting of microparticles, gels, bioadhesives, hydrogels, xerogels, foams and combinations
thereof. In one embodiment, said medium comprises a bocompatible material. In one
embodiment, said medium comprises a biodegradable material. In one embodiment, said
medium provides controlled release of said compound. In one embodiment, said
microparticles are selected from the group consisting of microspheres, microencapsulating
particles, microcapsules and liposomes. In one embodiment, the microparticle comprises a
polymer selected from the group consisting of poly(lactidle-co-glycolide), aliphatic polyesters
including, but not limited to, poly-glycolic acid and poly- lactic acid, hyaluronic acid, modified
polysacchrides, chitosan, cellulose, dextran, polyurethanes, polyacrylic acids, psuedo-
poly(amino acids), polyhydroxybutrate-related copolymers, polyanhydrides,
polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids. In one embodiment
the medium comprises a material selected from the group consisting of gelatin, collagen,
cellulose esters, dextran sulfate, pentosan polysulfate, chitin, saccharides, albumin, fibrin

sealants, synthetic polyvinyl pyrrolidone, polyethylene oxide, polypropylene oxide, block
i
polymers of polyethylene oxide and polypropylene oxide, (polyethylene glycol, acrylates,
acrylamides, methacrylates including, but not limited to, 2-hydroxyethyl methacrylate,
poly(ortho esters), cyanoacrylates, gelatin-resorcin-aldehyde type bioadhesives, polyacrylic
acid and copolymers and block copolymers thereof. In another embodiment, the medium
comprises a polymer selected from the group consisting of poly(lactide-co-glycolide), aliphatic
polyesters including, but not limited to, poly-glycolic acid and poly-lactic acid, hyaluronic
acid, modified polysacchrides, chitosan, cellulose, dextranj polyurethanes, polyacrylic acids,
psuedo-poly(amino acids), polyhydroxybutrate-related copplymers, polyanhydrides,
polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids. In one embodiment,
said medium is colored. In one embodiment, said medium further comprises a radio-opaque
marker, wherein said marker is visualized by X-ray spectoscopy. In one embodiment, said
analog of sirolimus is selected from the group consisting of everolimus, CCI-779, ABT-578,
7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin,
7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin and
2-desmethyl-rapamycin. In one embodiment, said medium further comprises a supplemental
pharmaceutical compound selected from the group consisting of antiinflammatory,
corticosteriods, antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
One aspect of the present invention contemplates a composition, comprising:
a) a medium; and b) a compound attached to said medium, said compound selected from the
group consisting of sirolimus, tacrolimus, analogs of siro|imus and pharmaceutically
acceptable salts thereof. In one embodiment, said medium comprises a biocompatible
material. In another embodiment, said medium comprises a biodegradable material. In one
embodiment, said medium provides controlled release of said compound. In one embodiment,
said medium is selected from the group consisting of a raicroparticles, liquids, foams, gels,
hydrogels, xerogels and bioadhesives. In another embodiment, said medium is a spray. In
one embodiment, said medium comprises a microparticle. In one embodiment, said
microparticle is a microencapsulating particle. In one embodiment, said microencapsulating
particle is selected from the group consisting of microcapsules, microspheres and liposomes.
In one embodiment, said medium is colored. In one embodiment, said medium further

comprises a radio-opaque marker, wherein said marker is visualized by X-ray spectroscopy.
In one embodiment, said analog of sirolimus is selected from the group consisting of
everolimus, CCI-779, ABT-578, 7-epi-rapamycin, 7-thiomethyl-rapamycin,
7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rap4mycin, 7-demethoxy-rapamycin,
32-demethoxy-rapamycin and 2-desmethyl-rapamycin. Ifi one embodiment, said composition
further comprises antisense to c-myc. In another embodiment, said composition further
comprises tumstatin. In one embodiment, said composition further comprises a supplemental
pharmaceutical compound selected from the group consisting of antiinflammatory,
corticosteriods, antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
Another aspect of the present invention contemplates a composition, comprising:
a) a micfoparticle; and b) a compound attached to said tfiicroparticle, said compound selected
from the group consisting of sirolimus, tacrolimus, analogs of sirolimus and pharmaceutically
acceptable salts thereof. In one embodiment, said micrc)particle comprises a biocompatible
material. In one embodiment, said microparticle comprises a biodegradable material. In one
embodiment, said microparticle is a microsphere. In one embodiment, said microparticle is a
microencapsulating particle. In one embodiment, said medium provides controlled release of
said compound. In one embodiment, said microencapsulating particle is selected from the
group consisting of microcapsules and liposomes. In oite embodiment,, said microparticle is
colored. In one embodiment, said microparticle further, comprises a radio-opaque marker,
wherein said marker is visualized by X-ray spectroscopy. In one embodiment, said analog of
sirolimus is selected from the group consisting of everojimus, CCI-779, ABT-578,
7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimetbJoxyphenyl-rapamycin,
7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 3t2-demethoxy-rapamycin and
2-desmethyl-rapamycin. In one embodiment, said composition further comprises antisense to
c-myc. In another embodiment, said composition furtbjer comprises tumstatin. In one
embodiment, said composition further comprises, a supplemental pharmaceutical compound
selected from the group consisting of antiinflammatory^ corticosteriods, antithrombotics,
antibiotics, antivirals, analgesics and anesthetics.
One aspect of the present invention contemplates a composition, comprising: a) a
microparticle, wherein said microparticle encapsulates ja compound selected from the group

consisting of sirolimus, tacrolimus, analogs of sirolimus and pharmaceutically acceptable salts
thereof; and b) a biocompatible and biodegradable material to which said microparticle is
attached. In one embodiment, said microparticle is selected from the group consisting of
microspheres, microcapsules and liposomes. In one embodiment, said microparticle provides
controlled release of said compound. In one embodiment said microparticle is clear. In
another embodiment, said microparticle is colored. In one embodiment, said microparticle
further comprises a radio-opaque marker, wherein said marker is visualized by X-ray
spectroscopy. In one embodiment, said biocompatible and biodegradable material is selected
from the group consisting of polylactide-polyglycolide polymers, lactide/glycolide copolymers,
poly(lactide-co-glycolide) polymers (i.e., PLGA), hyaluropic acid, modified polysaccharides
and any other well known substance that is known to be both biocompatible and
biodegradable. In one embodiment, said analog of sirolimus comprises a compound capable
of binding to the mTOR protein. In one embodiment, sajd compound capable of binding to
the mTOR protein is selected from the group consisting of everolimus, CCI-779, ABT-578,
7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin,
7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-
rapamycin. In one embodiment, said composition further comprises anti-sense to c-myc. In
another embodiment, said composition further comprises tumstatin. In one embodiment, said
microparticle further comprises a plurality of supplemental pharmaceutical compounds. In
one embodiment, said supplemental pharmaceutical compound is selected from the group
consisting of antiinflammatory, corticosteriods, antithromtyotics, antibiotics, antivirals,
analgesics and anesthetics.
Another aspect of the present invention is a composition, comprising:
a) a biocompatible and biodegradable hydrogel; and b) a compound selected from the group
consisting of sirolimus, tacrolimus, analogs of sirolimus ahd pharmaceutically acceptable salts
thereof, wherein said compound is attached to said hydrogel. In one embodiment, said
hydrogel comprises a material selected from the group consisting of gelatins, pectins,
collagens, hemoglobins, carbohydrates, hyaluronic acid, cellulose esters, Carbopol®, synthetic
polyvinylpyrrolidone, polyethyleneoxide, acrylate, and mefthacrylate and copolymers thereof.
In one embodiment, said hydrogel provides controlled release of said compound. In one

embodiment, said analog of sirolimus comprises a compound capable of binding to the mTOR
protein. In one embodiment, said compound capable jof binding to the mTOR protein is
selected from the group consisting of everolimus, CCl-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapa|tnycin, 7-epi-thiomethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin. In one
embodiment, said composition further comprises antsense c-myc. In another embodiment,
said composition further comprises tumstatin. In one embodiment, said biodegradable and
biocompatible hydrogel further comprises a plurality of supplemental pharmaceutical
compounds. In one embodiment, said supplementall pharmaceutical compound is selected
from the group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics. In one embodiment, a cytostatic pharmaceutical
compound is attached to a polymer medium that is incorporated into said hydrogel. In one
embodiment, said polymer medium is biodegradable and has a different release rate and
biodegradation characteristics than said hydrogel. In another embodiment, said polymer
medium is selected from the group comprising polylactide-polyglycolide polymers,
lactide/glycolide copolymers, poly(lactide-co-glycolide) polymers (i.e., PLGA), hyaluronic
acid or other similar polymers. In one embodiment, said hydrogel comprises a microparticle
incorporating a cytostatic drug,
Another aspect of the present invention contemplates a composition, comprising:
a) a biocompatible bioadhesive; and b) a compound selected from the group consisting of
sirolimus, everolimus, analogs of sirolimus and pharmaceutically acceptable salts thereof,
wherein said compound is attached to said bioadhesive. In one embodiment, said bioadhesive
is biodegradable. In one embodiment, said bioadhesive provides controlled release of said
compound. In one embodiment, said bioadhesive comprises a material selected from the
group consisting of fibrin, fibrinogen, calcium polycarbophil, polyacrylic acid, gelatin,
carboxymethyl cellulose, natural gums such as karaya and tragacanth, algin, cyanoacrylates,
chitosan, hydroxypropylmethyl cellulose, starches, pectins or mixtures thereof. In one
embodiment, said bioadhesive further comprises a hydrocarbon gel base, wherein said base is
composed of polyethylene and mineral oil. In one embodiment, said base has a preselected
pH level, wherein said pH level maintains, said jbase stability. In one embodiment, said analog

of sirolimus comprises a compound capable of binding to the mTOR protein selected from the
group consisting of tacrolimus, everolimus, CCI-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desm)5thyl-rapamycin. In one
embodiment, said composition further comprises antisense to c-myc. In another embodiment,
said composition further comprises tumstatin. In one embodiment, said bioadhesive further
comprises a plurality of supplemental pharmaceutical compounds. In one embodiment, said
supplemental pharmaceutical compound is selected from the group consisting of
antiinflammatory, corticosteriods, antitlirombotics, antibiotibs, antivirals, analgesics and
anesthetics.
Another aspect of the present invention contemplatejs a gel, comprising a compound
selected from the group consisting of sirolimus, everolimus, analogs of sirolimus and
pharmaceutically acceptable salts thereof. In one embodimjent, said gel comprises a hydrogel.
In one embodiment, said gel provides controlled release of said compound. In one
embodiment, said gel is colored. In one embodiment, said gel further comprises a radio-
opaque marker, wherein said marker is visualized by X-ray) spectroscopy. In one
embodiment, said analog of sirolimus is selected from the group consisting of everolimus,
CCI-779, ABT-578, 7-epi-rapamycin, 7-thiomethyl-rapamypin,
7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin,
32-demethoxy-rapamycin, 2-desmethyl-rapamycin. In one jembodiment, said gel further
comprises antisense c-myc. In another embodiment, said g-el further comprises tumstatin. In
one embodiment, said gel further comprises a supplemental pharmaceutical compound selected
from the group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics. One embodiment contemplates a surgical device
wherein at least a portion of said device comprises an attached gel comprising sirolimus and
analogs of sirolimus.
Another aspect of the present invention contemplates a foam, comprising a compound
selected from the group consisting of sirolimus, tacrolimus, analogs of sirolimus and
pharmaceutically acceptable salts thereof. In one embodiment, said foam further comprises a
xerogel. In one embodiment, said foam provides controlled release of said compound. In one

embodiment, said foam is colored. In one embodiment, said foam further comprises a radio-
opaque marker, wherein said marker is visualized by X-raV spectroscopy. In one
embodiment, said analog of sirolimus is selected from the group consisting of everolimus,
CCI-779, ABT-578, 7-epi-rapamyein, 7-thiomethyl-rapamycin,
7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-raparmycm, 7-demethoxy-rapamycin,
32-demethoxy-rapamycin, 2-desmethyl-rapamycin. In one I embodiment, said foam further
comprises antisense c-myc. In another embodiment, said from further comprises tumstatin.
In one embodiment, said foam further comprises a supplemental pharmaceutical compound
selected from the group consisting of antiinflammatory, corticosteriods, antithrombotics,
antibiotics, antivirals, analgesics and anesthetics. One embodiment contemplates a surgical
device wherein at least a portion of said device comprises an attached foam comprising
sirolimus and analogs of sirolimus.
One aspect of the present invention contemplates a method, comprising: a) providing:
i) a medium comprising a compound selected from the group consisting of sirolimus,
tacrolimus, analogs of sirolimus and pharmaceutically acceptable salts thereof, wherein said
medium is selected from the group consisting of micropartilcles, gels, xerogels, hydrogels,
bioadhesives, foams and combinations thereof; and ii) a patient, wherein said patient has a
surgical site; and b) contacting said surgical site with said medium. In one embodiment, said
surgical site comprises a closed surgical site. In another embodiment, said surgical site
comprises an open surgical site. In one embodiment, said medium of step a) is housed in a
device capable of delivering said medium to said surgical site. In One embodiment, said
device delivers said medium by brushing. In one embodiment, said device delivers said
medium by liquid administration. In one embodiment, said liquid administration comprises a
liquid spray. In one embodiment, said liquid spray in the form of an aerosol. In one
embodiment, said liquid administration comprises a pourable liquid. In another embodiment,
said liquid administration comprises a squeezable liquid. In one embodiment, said device
comprises a catheter. In one embodiment, said device is configured for endoscopic surgery.
In one embodiment, said medium comprises a biocompatible material. In one embodiment,
said medium comprises a biodegradable material. In one embodiment, said microparticles are
selected from the group consisting of microspheres, microencapsulating particles,

microcapsules and liposomes. In one embodiment, said medium is colored. In one
embodiment, said medium further comprises a radio-opaque marker, wherein said marker is
visualized by X-ray spectroscopy. In one embodiment, said analog of sirolimus is selected
from the group consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycm, 7-epi-thipmethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin and 2-desmethyl-rapamycin. In one
embodiment, said method further comprises administering antisense to c-myc. In another
embodiment, said method further comprises administering tumstatin. In one embodiment, said
medium further comprises administering a supplemental pharmaceutical compound selected
from the group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics. In one embodiment, said method further comprises
administering a complementary pharmaceutical compound selected from the group consisting
of sirolimus, tacrolimus, analogs of sirolimus, antiinflammatory, corticosteriods,
antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
One aspect of the present invention contemplates a method, comprising: a) providing:
i) a medium comprising a compound selected from the group consisting of sirolimus,
tacrolimus, analogs of sirolimus and pharmaceutically acceptable salts thereof, wherein said
medium is selected from the group consisting of microparticles, gels, xerogels, hydrogels,
bioadhesives, foams and combinations thereof; and ii) a patient, wherein said patient has a
wound; and b) contacting said wound with said medium. In one embodiment, said wound is
external. In another embodiment, said wound is internal. In one embodiment, said medium
of step a) is housed in a device capable of delivering said medium to said wound. In one
embodiment, said device delivers said medium by brushing. In one embodiment, said device
delivers said medium by liquid administration. In one embodiment, said liquid administration
comprises a liquid spray. In one embodiment, said liquid spray in the form of an aerosol. In
one embodiment, said liquid administration comprises a pourable liquid. In another
embodiment, said liquid administration comprises a squeezable liquid. In one embodiment,
said device comprises a catheter. In one embodiment, said device is configured for
endoscopic surgery. In one embodiment, said medium comprises a biocompatible material.
In one embodiment, said medium comprises a biodegradable material. In one embodiment,

said microparticles are selected from the group consisting of microspheres, microencapsulating
particles, microcapsules and liposomes. In one embodiment, said medium is colored. In one
embodiment, said medium further comprises a radio-opaque marker, wherein said marker is
visualized by X-ray spectroscopy. In one embodiment, said analog of sirolimus is selected
from the group consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin and 2-desmethyl-rapamycin. In one
embodiment, said method further comprises administering antisense to c-myc. In another
embodiment, said method further comprises administering tumstatin. In one embodiment, said
medium further comprises administering a supplemental pharmaceutical compound selected
from the group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics. In one embodiment, said method further comprises
administering a complementary pharmaceutical compound selected from the group consisting
of sirolimus, tacrolimus, analogs of sirolimus, antiinflammatory, corticosteriods,
antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
Another aspect of the present invention contemplates a method, comprising:
a) providing: i) a composition, comprising a medium and a compound attached to said
medium, said compound selected from the group consisting of sirolimus, tacrolimus, analogs
of sirolimus and pharmaceutically acceptable salts thereof; and ii) a patient, wherein said
patient has a surgical site; and b) contacting said surgical site with said composition. In one
embodiment, said surgical site comprises a closed surgical site. In one embodiment, said
composition of step a) is housed in a device comprising a reservoir, wherein said device is
capable of delivering said composition to a surgical site. In one embodiment, said medium
comprises a biocompatible material. In one embodiment, said medium comprises a
biodegradable material. In one embodiment, said medium is selected from the group
consisting of microparticles, gels, xerogels, hydrogels, bioadhesives, foams and combinations
thereof. In another embodiment, said medium provides controlled release of said compound.
In one embodiment, said microparticles are microencapsulating particles. In one embodiment,
said microencapsulating particle is selected from the group consisting of microcapsules and
liposomes. In one embodiment, said composition of step a) contacts said surgical site in the

form of a spray. In one embodiment, said device delivers said composition by brushing. In
one embodiment, said device delivers said medium by liquid administration. In one
embodiment, said liquid administration comprises a liquid spray. In one embodiment, said
liquid spray in the form of an aerosol. In one embodiment, said liquid administration
comprises a pourable liquid. In another embodiment, said liquid administration comprises a
squeezable liquid. In one embodiment, said device comprises a catheter. In one embodiment,
said method further comprises observing said contacting of said surgical site with an
endoscopic device. In another embodiment, said method further comprises observing said
contacting of said surgical site with a fiuoroscopic device. In one embodiment, medium is
colored. In one embodiment, said medium further comprises a radio-opaque marker, wherein
said marker is visualized by X-ray spectroscopy. In one embodiment, said analog of sirolimus
is selected from the group consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-raparnycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin and 2-desmethyl-rapamycin. In one
embodiment, said method further comprises administering antisense to c-myc. In another
embodiment, said method further comprises administering tumstatin. In one embodiment, said
medium further comprises administering a supplemental pharmaceutical compound selected
from the group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics. In one embodiment, said method further comprises
administering a complementary pharmaceutical compound selected from the group consisting
of sirolimus, tacrolimus, analogs of sirolimus, antiinflammatory, corticosteriods,
antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
Another aspect of the present invention contemplates a method, comprising:
a) providing: i) a composition, comprising a microparticle and a compound attached to said
microparticle, said compound selected from the group consisting of sirolimus, tacrolimus
analogs of sirolimus, and pharmaceutically acceptable salts thereof; and ii) a patient, wherein
said patient has a surgical site; and b) contacting said surgical site with said composition. In
one embodiment, said surgical site comprises a closed surgical site. In another embodiment,
said surgical site comprises an open surgical site. In one embodiment, said composition of
step a) is housed in a device comprising a reservoir, wherein said device is capable of

delivering said composition to a surgical site. In one embodiment, said device delivers said
composition by brushing. In one embodiment, said device delivers said composition by liquid
administration. In one embodiment, said liquid administration comprises a liquid spray. In
one embodiment, said liquid spray in the form of an aerosol. In one embodiment, said liquid
administration comprises a pourable liquid. In another embodiment, said liquid administration
comprises a squeezable liquid. In one embodiment, said device comprises a catheter. In one
embodiment, said method further comprises observing said contacting of said surgical site
with an endoscopic device. In another embodiment, said method further comprises observing
said contacting of said surgical site with a fluoroscopic device. In one embodiment, said
microparticle comprises a biocompatible material. In one embodiment, said microparticle
comprises a biodegradable material. In one embodiment, said microparticle is a microsphere.
In one embodiment, said microparticle is a microencapsulating particle. In one embodiment,
said microencapsulating particle is selected from the group consisting of microcapsules and
liposomes. In one embodiment, said microparticle is colored. In one embodiment, said
microencapsulating particle further comprises a radio-opaque marker, wherein said marker is
visualized by X-ray spectroscopy. In one embodiment, said analog of sirolimus is selected
from the group consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin,
7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-rapamycin. In one
embodiment, said method further comprises administering antisense to c-myc. In another
embodiment, said method further comprises administering tumstatin and antisense c-myc. In
one embodiment, said medium further comprises administering a supplemental pharmaceutical
compound selected from the group consisting of antiinflammatory, corticosteriods,
antithrombotics, antibiotics, antivirals, analgesics and anesthetics. In one embodiment, said
method further comprises administering a complementary pharmaceutical compound selected
from the group consisting of sirolimus, tacrolimus, analogs of sirolimus, antiinflammatory,
corticosteriods, antithrombotics, antibiotics, antivirals, analgesics and anesthetics.
Another aspect of the present invention contemplates a method comprising:
a) providing; i) a patient, wherein said patient has an open surgical site; ii) a biocompatible
medium, wherein said medium is attached to a compound selected from the group consisting

of sirolimus, tacrolimus, analogs of sirolimus and pharmaceutical acceptable salts thereof; and
iii) a medical device containing said medium, wherein said medical device is capable of
administering said compound to said surgical site; b) contacting said surgical site with said
medium by administering said medium from said medical device; and c) reducing the
formation of excess post-operative scar tissue and/or adhesions by pharmacological activity of
said compound. In one embodiment, said medium is biodegradable. In one embodiment, said
medium provides controlled release administration of sirolimus or analogs of sirolimus. In
one embodiment, said medium comprises a microencapsulating particle. In another
embodiment, said medium is selected from the group consisting of a gel, foam, dressing and
bioadhesive. In one embodiment, said compound contacts said surgical site by liquid
administration. In one embodiment, said contacting is selected from the group consisting of a
spraying, brushing, wrapping and layering. In one embodiment, said microencapsulating
particle is selected from the group consisting of microparticles, microspheres, microcapsules
and liposomes. In one embodiment, said medium is comprised of a material selected from the
group consisting of polylactide-polyglycolide polymers, lactide/glycolide copolymers,
poly(lactide-co-glycolide) polymers (i.e., PLGA), hyaluronic acid, modified polysaccharides
and any other well known substance that is known to be both biocompatible and
biodegradable. In one embodiment said analog of sirolimus comprises a compound capable of
binding to the mTOR protein selected from the group consisting of everolimus, CCI-779,
ABT-578, 7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin,
7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin and
2-desmethyl-rapamycin. In one embodiment, said method further comprises administering
antisense to c-myc. In another embodiment, said method further comprises administering
tumstatin. In one embodiment, said medical device is selected from the group consisting of a
self-contained spray container, a gas-propelled spray container, a spray catheter, a liquid-
dispensing catheter, a brush, and a syringe. In one embodiment, said spray can comprises a
single dose of said compound. In one embodiment, said spray can comprises a
microencapsulating particle contacting said compound. In one embodiment, said medium is
colored. In one embodiment, said medium further comprises a radio-opaque marker, wherein
said marker is visualized by X-ray fluoroscopy. In one embodiment, said method further

comprises administering a supplemental pharmaceutical compound selected from the group
consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics, antivirals,
analgesics and anesthetics. In one embodiment, said method further comprises administering
a complementary pharmaceutical compound selected from the group consisting of sirolimus,
tacrolimus, analogs of sirolimus, antiinflammatory, corticosteriods, antithrombotics,
antibiotics, antivirals, analgesics and anesthetics. In one embodiment, administration of said
complementary pharmaceutical compound starts prior to exposure of said surgical site a
surgical procedure. In another embodiment, administration of said complementary
pharmaceutical compound continues for up to 6 months following exposure of said surgical
site.
Another aspect of the present invention contemplates a method comprising:
a) providing; i) a patient, wherein said patient has a closed surgical site; ii) a biocompatible
medium, wherein said medium is attached to a compound selected from the group consisting
of sirolimus, tacrolimus, analogs of sirolimus and pharmaceutically acceptable salts thereof;
and iii) a medical device containing said medium, wherein said medical device is capable of
administering said medium to said surgical site; b) contacting said surgical site with said
medium by administering said medium from said medical device; and c) reducing formation
of excess post-operative scar tissue and/or adhesions by pharmacological activity of said
compound. In one embodiment, said medium is biodegradable. In one embodiment, said
method further comprises a step of, visualizing said surgical site with an endoscope to guide
and verify said medium administration. In one embodiment, said analog of sirolimus
comprises a compound capable of binding to the mTOR protein selected from the group
consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin, 7-thiomethyl-rapamycin,
7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin,
32-demethoxy-rapamycin and 2-desmethyl-rapamycin. In one embodiment, said medium
further comprises antisense to c-myc. In another embodiment, said medium further comprises
tumstatin. In one embodiment, said medical device is selected from the group consisting of a
catheter and said endoscope. In one embodiment, said catheter is capable of layering said
medium. In one embodiment, said catheter is capable of spraying said medium. In one
embodiment, said catheter is capable of liquid administration of said medium. In another

embodiment, said catheter is capable of brushing said medium. In one embodiment, said
catheter pours said medium. In another embodiment, said medium is selected from the group
consisting of a microparticle, foam, gel, hydrogel, liquid spray and bioadhesive. In one
embodiment, said method further comprises administering a supplemental pharmaceutical
compound selected from the group consisting of antiinflammatory, corticosteriods,
antithrombotics, antibiotics, antivirals, analgesics and anesthetics. In one embodiment, said
method further comprises administering a complementary pharmaceutical compound selected
from the group consisting of sirolimus, tacrolimus, analogs of sirolimus, antiinflammatory,
corticosteriods, antithrombotics, antibiotics, antivirals, analgesics and anesthetics. In one
embodiment, administration of said complementary pharmaceutical compound starts prior to
exposure of said surgical site a surgical procedure. In another embodiment, administration of
said complementary pharmaceutical compound continues for up to 6 months following
exposure of said surgical site.
One aspect of the present invention contemplates a device, comprising: i) a reservoir
containing a medium comprising sirolimus and analogs of sirolimus; ii) a fluid-driving
element connected to said reservoir; iii) a channel having a first end and a second end,
wherein said first end is connected to said reservoir; and iv) an extrusion port located at the
second end of said channel, whereby said fluid-driving element causes said medium to extrude
from said extrusion port.
One aspect of the present invention contemplates a device, said device comprising a
reservoir comprising a medium comprising sirolimus and analogs of sirolimus and capable of
delivering said medium to a surgical site. In one embodiment, said delivering is in the form
of a spray. In one embodiment, said delivering is in the form of an aerosol. In one
embodiment, said device comprises a catheter. In one embodiment, said device is an
endoscope. In one embodiment, said endoscope is a laparoscope. One embodiment
contemplates a surgical device wherein at least a portion of said device comprises an attached
medium comprising sirolimus and analogs of sirolimus.
Another aspect of the present invention contemplates a device, said device comprising
a reservoir comprising sirolimus and analogs of sirolimus and is capable of delivering said
sirolimus and analogs of said sirolimus to a surgical site. In one embodiment, said delivering

is in the form of a spray. In one embodiment, said delivering is in the form of an aerosol.
In one embodiment, said device comprises a catheter. In one embodiment, said device
comprises a laparoscopic device. In one embodiment, said device is a surgical device wherein
at least a portion of said device is coated with sirolimus and analogs of sirolimus.
These and other embodiments and applications of this invention will become obvious
to a person of ordinary skill in this art upon reading of the detailed description of this
invention including the associated drawings.
Brief Description Of The Drawings
Figure 1 illustrates one embodiment of a liposome encapsulating a sirolimus molecule.
Figure 2 illustrates one embodiment of a microsphere impregnated with a cytostatic
anti-proliferative compound.
Figure 3 illustrates one embodiment of a microsphere to which a cytostatic
anti-proliferative compound is adhered to the surface.
Figure 4 illustrates one embodiment of a microsphere comprising controlled release of
sirolimus or an analog of sirolimus.
Figure 5 illustrates one embodiment of a spray can to administer a sirolimus medium.
Figure 6 shows one embodiment of a nebulizer tip attached to a syringe.
Figure 7 illustrates by cross-section one embodiment of an endoscope shaft containing
a endoscopic catheter delivering a medium.
Figure 8 shows one embodiment of an endoscopic catheter for administration of media.
Note: typical sizes are length = 200 cm; diameter = 2.5 mm and length of area 3 (w/ holes) =
4 cm. Note: Female luer lock 81 that readily fits onto a syringe or other plunger device or
spray can.
Figure 9 shows one embodiment of a foam canister.
Figure 10 shows one embodiment of surgical dressing.
Figure 11 shows two exemplary embodiments of a side hole catheter.
Figure 12 shows a close-up view of one embodiment of a slit port spray catheter tip.
Figure 13 shows one embodiment of a bioadhesive applicator.

Detailed Description of The Invention
This invention is related to the field of tissue healing and reducing scar tissue
formation. More specifically, this invention is related to the use of sirolimus and analogs of
sirolimus (i.e., sirolimus and it's derivatives) to reduce post-surgical scar tissue formation.
This invention also contemplates the use of compounds that are capable of binding to the
mTOR protein to reduce and/or prevent scar tissue formation. The binding of compounds to
the mTOR protein may be direct or indirect, competitive or non-competitive. Also
contemplated are allosteric agonists or antagonists that may increase or decrease, respectively,
the binding efficacy of a compound to the mTOR protein. Also contemplated by this
invention are embodiments of antiproliferative, cytostatic compounds (i.e., sirolimus and
analogs of sirolimus) which are believed to act primarily by interrupting the cell division
cycle at the GO or Gl phase in such a way that cell death does not occur.
Sirolimus and its derivatives is currently marketed as an antiproliferative, cytostatic
drug in the liquid form for oral administration in 1 - 5 dosages per day of between 1-100
mg each. These disclosed oral mediums consist of conventional tablets, capsules, granules
and powders. Guitard et al., Pharmaceutical Compositions. United States Patent No.
6,197,781 (herein incorporated by reference).
Until recently, none of the clinical uses for the above liquid sirolimus compositions
had been contemplated for the reduction of excess scar tissue. Sirolimus (i.e., rapamycin) is
useful for treatment of post-surgical adhesions and scar tissue, wherein the drug is attached to
a sheet of material and placed onto a damaged area. As the sirolimus is released from the
sheet of material, it exerts it antiproliferative action. Fischell et al, Surgically Implanted
Devices Having Reduced Scar Tissue Formation, United States Patent No. 6,534,693 (2003).
One aspect of the present invention contemplates delivering sirolimus or other
cytostatic agents to a surgical site or wound in a controlled release manner {i.e., ranging from
1 day to 6 months). In some embodiments described herein, a specific medium is
contemplated comprising specific formulations of polymers that provide a controlled drug
release capability where the polymers take the form of microparticles, gels, foams or liquids..
In one embodiment, a local administration of a cytostatic compound is administered

concurrently with a systemic administration of said cytostatic compound.
Another aspect of the present invention contemplates a variety of devices and methods
to administer a medium comprising an attached compound. Preferably, these devices and
methods include, but are not limited to, spray cans, reservoirs with plungers, delivery via a
catheter for endoscopic procedures, premixed media, and media mixed at the time of
administration.
Sirolimus and most analogs of sirolimus are known not to be readily soluble in
aqueous solutions. A non-polar solvent or amphipathic material is usually required to
generate a liquid solution (i.e., for example, olive oil). Otherwise, aqueous mixtures of
sirolimus and analogs of sirolimus are limited to colloidal suspensions or dispersions. The
present invention contemplates a method to improve the solubility of sirolimus. To this end,
modified derivatives of sirolimus are contemplated in order to address this problem.
Soluble monoacyl and diacyl derivatives of sirolimus can be prepared according to
known methods. Rakhit, U.S. Pat. No. 4,316,885 (herein incorporated by reference). These
derivatives are used in the form of a sterile solution or suspension containing other solutes or
suspending agents, for example, enough saline or glucose to make the solution isotonic, bile
salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitol and its
anhydrides copolymerized with ethylene oxide) and the like. Furthermore, water soluble
prodrugs of sirolimus may be used including, but not limited to, glycinates, propionates and
pyrrolidinobutyrates. Stella et al., U.S. Pat. No. 4,650,803 (herein incorporated by reference).
Alternatively, aminoalkylation of sirolimus or analogs of sirolimus to create functional
sirolimus derivatives is contemplated. Kingsbury et al. Synthesis Of Water-Soluble
(Aminoalkyl) camptothecin Analogues: Inhibition Of Topoisomerase I And Antitumor Activity,
J. Med. Chem. 34:98(1991). The Kingsbury et al. publication teaches the synthesis of
several water-soluble analogs of camptothecin, by introduction of atninoalkyl groups into the
camptothecin ring system. These derivatives, retained their biological efficacy.
Alternatively, sirolimus or its analogs modified by bonding phenolic groups with
diamines through a monocarbamate linkage is contemplated as having improved solubility.
For example, it is known that the water solubility of camptotecin is improved by derivatives

bonding to phenolic groups with diamines through a monocarbamate linkage. Sawada et al,
Synthesis And Antitumor Activity Of 20(S)-Camptothecin Derivatives: Carbamate-Linked,
Water Soluble Derivatives Of'7-Ethyl-10-hydroxycamptothecin, Chem. Pharm. Bull 39:1446
(1991).
It is known that 6eto-emitting radioisotopes placed onto a sheet of material reduce scar
tissue formation. Although effective, the limited shelf life and safety issues associated with
clinical use of radioisotopes make them less than ideal for routine use in the operating room
or a doctor's office. Fischell et al., U.S. Patent. No. 5,795,286 (herein incorporated by
reference).
Various means and methods to reduce scar tissue formation are disclosed in the art, but
none utilizing the pharmacological activity of a cytostatic compound. For example, sheets of
biodegradable mesh, gels, foams and barrier membranes of various materials, are
commercially available or in clinical trials that are intended to reduce unwanted scar tissue
growth and post-surgical adhesions. The mechanism of action of these barrier membranes is
not pharmacological but involves a physical separation of the injured tissues, thereby
preventing adherence.
ACTIONS OF SIROLIMUS AND RELATED COMPOUNDS
The present invention contemplates the administration of cytostatic, anti-proliferative
compounds such as, but not limited to, sirolimus, tacrolimus (FK506) and any analog of
sirolimus including, but not limited to everolimus (i.e., SDZ-RAD), CCI-779, ABT-578,
7-epi-rapamycin, 7-thiomethyl-rapamycin, 7-epi-trimethoxyphenyl-rapamycin,
7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-demethoxy-rapamycin, 2-desmethyl-
rapamycin. In one embodiment, the present invention contemplates non-sirolimus compounds
such as, but not limited to, antisense to c-myc (Resten-NG) and tumstatin.
Inhibition of mTOR
Cytostatic antiproliferative compounds, such as sirolimus (i.e. sirolimus) and its
functional analogs are known to reduce cell proliferation. Originally discovered as an
antifungal agent, the bacterial macrolide sirolimus is a potent immunosuppressant, a promising

anti-cancer compound and an antiproliferative compound. Although it is not necessary to
understand the mechanism of an invention, it is believed that sirolimus forms a complex with
its cellular receptor, the FK506-binding protein (FKBP12), and inhibits the function of
mammalian Target Of Rapamycin (mTOR). Current understanding indicates that by
mediating amino acid sufficiency, mTOR governs signaling to translational regulation and
other cellular functions by converging with the phosphatidylinositol 3-kinase pathway on
downstream effectors. Recent findings have revealed a novel link between mitogenic signals
and mTOR via the lipid second messenger phosphatidic acid that suggests mTOR may be
involved in the integration of nutrient and mitogen- signals. One hypothesis suggests that this
possible interaction between phosphatidic and mTOR is inhibited by sirolimus binding. Chen
et al, A Novel Pathway Regulating The Mammalian Target Of Sirolimus (mTOR) Signaling.
Biochem Pharmacol. 64:1071-1077 (2002).
The binding of sirolimus, or sirolimus analogs, to the mTOR protein may be direct or
indirect, or depend upon the binding of facilitating compounds, such as, allosteric agonists.
Conversely, the binding of sirolimus or sirolimus analogs to the mTOR protein may depend
on the binding of inhibiting compounds, such as, allosteric antagonists. Consequently, one of
skill in the art would understand that the resultant change in mTOR protein activity due to the
presence of sirolimus, or analogs of sirolimus, may not be solely dependent upon binding to
sirolimus, or analogs of sirolimus.
Cell Cycle Interruption
Although it is not necessary to understand the mechanism of an invention, it is
believed that the principal action of cytostatic antiproliferatives such as, sirolimus and analogs
of sirolimus, is an interference with the progress of the cell cycle at the GO or G1 phase.
Other compounds capable of binding to the mTOR protein are also expected to decrease
cellular proliferation and hence reduce the formation of excess scar tissue at a surgical or
epidermal wound site. Compounds capable of binding to the mTOR protein may or may not
have structure similarity to sirolimus or analogs of sirolimus nor may they have similar
mTOR binding sites. Other cytostatic antiproliferatives that interfere with the GO or Gl phase
of the cell cycle are also contemplated within this invention to effectively reduce scar tissue
when properly dispensed to a surgical or other injury site.

Sirolimus and its analogs impact a variety of cell types. In the case of preventing
vascular hyperplasia following angioplasty it is believed that the dominant mechanism of
sirolimus released at the site of a vascular stent is to inhibit growth factor and cytokine
mediated smooth muscle cell proliferation at the Gl phase of the cell cycle. In its application
as an anti-rejection drug, sirolimus is administered systemically to prevent T-cell proliferation
and differentiation. Moses, J.W., Brachytherapy And Drug Eluting Stents, J Invasive
Cardiology, 15:30B-33B (2003).
Glib-1 oncongeny expression occurs in both scar tissue and keloids, wherein keloids
express greater hyperproliferative characteristics, and glib-1 expression, than ordinary scar
tissue. Since sirolimus is known to inhibit glib-1 oncogeny expression it is expected that
sirolimus also inhibits glib-1 expression in keloids. Kim et al., Are Keloid Really "Glib-
loids": High-Level Expression Of Glib-1 Oncogeny In Keloid. J. Am Acad Dermatol
45(5):707-711 (2001). In one embodiment, the present invention contemplates a reduction in
keloid formation following the administration of sirolimus or analogs of sirolimus.
ACTIONS OF NON-SIROLIMUS RELATED COMPOUNDS
Cvtotoxic/Antiproliferative Compounds
Other cytotoxic compounds (i.e., taxol and other anticancer compounds), may or may
not bind to the mTOR protein, and have anti-proliferative effects, but are typically cytotoxic:
These cytotoxic compounds interfere with proliferation in part by interfering with successful
cell division in stage G2 or M, resulting in cell death. Because the by-products of cell death
are in and of themselves inflammatory and stimulative, it is believed that stopping cell
proliferation with a cytostatic effect rather than a cytotoxic effect is preferred. Indeed, in
drug coated stent trials, arteries treated with stents coated with a cytostatic drug (sirolimus)
showed less neointimal tissue growth than vessels treated with stents coated with a cytotoxic
drug (paclitaxel). Grube et al, Taxus I: Six And Twelve Month Results From A Randomized,
Double Blind Trial On Slow Release Paclitaxel Eluting Stent For De Novo Coronaiy Lesions.
Circulation 107:38-42 (2003); and Morice et al.A Randomized Comparison Of Sirolimus-
Eluting Agent With A Standard Stent For Coronary Revascularization. N Engl J Med

346:1773-1780 (2002).
The present invention also contemplates cytotoxic anti-proliferative non-sirolimus
compounds including, but not limited to, anticancer compounds such as taxol, actinomycin-D,
alkeran, cytoxan, leukeran, cis-platinum, BiCNU, adriamycin, doxorubicin, cerubidine,
idamycin, mithracin, mutamycin, fluorouracil, methotrexate, thioguanine, toxotere, etoposide,
vincristine, irinotecan, hycamptin, matulane, vumon, hexalin, hydroxyurea, gemzar, oncovin
and etophophos. Preferably, cytotoxic antiproliferative non-sirolimus compounds are used in
combination with sirolimus, tacrolimus and analogs of sirolimus. Alternatively, cytotoxic
antiproliferative non-sirolimus compounds may also be used alone.
Non-Sirolimus mTOR Binding
One embodiment of the present invention contemplates the reduction of excess scarring
by compounds capable of inhibiting the mTOR protein. One exemplary compound is
tumstatin, a 28-kilodalton fragment of type IV collagen that displays both anti-angiogenic and
proapoptotic activity. Tumstatin is known to function as an endothelial cell-specific inhibitor
of protein synthesis, however, there is no speculation in the art regarding any ability to reduce
excess scar tissue.. Although it is not necessary to understand an invention, it is believed that
tumstatin acts through aVp3integrin, inhibits focal adhesion kinase, phosphatidylinositol
3-kinase, protein kinase B, mTOR, and prevention of the dissociation of eukaryotic initiation
factor 4E protein (eIF4E) from 4E-binding protein 1.
CURRENT CLINICAL APPLICATIONS OF SIROLIMUS
Although there are several known uses of sirolimus, none of them include a
combination of sirolimus and a medium where the medium is in the form of a microsphere,
gel, liquid, bioadhesive or foam. Also none of the known uses contemplate using such a
composition to prevent excess scar tissue growth following injury. In such a form, the
sirolimus can be adminsitered to the wound site easily, without missing any portions of the
affected tissue.
Scar Tissue Reduction
Sirolimus (i.e., rapamycin) attached to a sheet of material is known as a useful
treatment of post-surgical treatment of adhesions and scar tissue. Fischell et ai, Surgically

Implanted Devices Having Reduced Scar Tissue Formation, United States Patent No.
6,534,693 (2003). Scarring, as used herein, also contemplates the narrowing of any
neurological, vascular, ductal/tubal (e.g., for example, pancreatic, biliary or fallopian) space in
the body secondary to injury from, for example, implants, trauma, surgery or system and local
disease/infections. In one embodiment, the present invention contemplates the administration
of sirolimus, tacrolimus and analogs of sirolimus to reduce scarring in compositions
comprising a medium including, but not limited to, a foam, gel, bioadhesive that may or may
not be attached to a dressing or medical device. Further, the present invention contemplates
the long term administration of sirolimus in the prevention of scar tissue formation by
compositions that provide controlled release of sirolimus or related compounds.
Transplantations
Sirolimus (i.e., Rapamune: Wyeth, Madison, NJ) is known as an immunosuppressant
effective for long-term immunosuppressive therapy in renal transplantation. Observations
indicate that sirolimus operates synergistically with cyclosporin A (CsA). For example, in
blinded dose-controlled trials, the rates of acute rejection episodes within 12 months following
administration of 2 or 5 mg/day sirolimus in combination with CsA and steroids were reduced
to 19 and 14%, respectively. It is speculated that sirolimus acts to retard proliferation of
vascular smooth muscle cells, an important component of the immuno-obliterative processes
associated with chronic rejection. Kahan, Sirolimus: A Comprehensive Review. Expert Opin
Pharmacother 2:1903-17 (2001).
The administration of mTOR inhibitors {i.e., sirolimus) are known to result in
improved outcomes for renal transplant recipients by decreasing the risk of rejection and by
increasing the function and lifespan of the allograft. Gourishankar et ai, New Developments
In Immunosuppressive Therapy In Renal Transplantation. Expert Opin Biol Ther 2:483-501
(2002)
Tacrolimus and sirolimus are two immunosuppressive compounds considered as
optimal immunosuppressive strategies for pancreas transplantation. Specifically, the
application of these compounds have contributed to substantially lower rates of allograft
rejection and improved graft survival. Odorico et al, Technical And Immunosuppressive

Advances In Transplantation For Insulin-Dependent Diabetes Mellitus. World J Surg
26:194-211 (2002). Similar effects on renal transplantation success has been reported
following the administration of an analog of sirolimus; SDZ RAD (everolimus, Certican®).
Nashan, Early Clinical Experience With A Novel Sirolimus Derivative. Ther Drug Monit
24:53-8 (2002).
Vascular Stents
Sirolimus is known as a coating for intraluminal vascular medical devices and methods
of treating intimal hyperplasia, constrictive vascular remodeling and resultant vascular scarring
and injury-induced vascular inflammation. Falotico et al., Compound/Compound Delivery
Systems For The Prevention And Treatment Of Vascular Disease. Published United States
Patent Application No. 2002/0007214 Al, Published United States Patent Application No.
2002/0007215 Al, United States Patent Application No. 2001/0005206 Al, United States
Patent Application No. 2001/007213 Al, United States Patent Application No. 2001/0029351
Al; and Morris et al., Method Of Treating Hyperproliferative Vascular Diseases. United
States Patent No. 5,665,728. These conditions are generally referred to as hyperproliferative
vascular disease and may be caused by vascular catheterization, vascular scraping,
percutaneous transluminal/coronary angioplasty, vascular surgery, vascular endothelial
proliferation, intimal hyperplasia, foreign body endothelial proliferation, and obstructive
proliferation/hyperplasia including specific conditions such as, but not limited to, fibroblastic,
endothelial or intimal.
Vascular stents have been coated with sirolimus (i.e., sirolimus), actinomycin-D or
taxol to reduce cellular proliferation and irestenosis following angioplasty or recanalization of
injured arteries. However, these compositions have never been used for reducing cellular
proliferation at the site of a surgical procedure. Hossainy et al., Process For Coating Stents.
United States Patent No. 6,153,252.
Sirolimus has been demonstrated to inhibit smooth muscle cell (SMC) proliferation
and migration in vitro and to reduce in vivo neointima formation by blocking the cell cycle
before the Gl-S transition. Further, sirolimus drug-eluting stents eliminate restenosis after
stent implantation. Paclitaxel (Taxol: a microtubule- stabilizing agent) has a similar

antiproliferative effect. Paclitaxel, however, is believed to act by inhibiting spindle formation
necessary for cell division. Chieffo et al, Drug-Eluting Stents. Minerva Cardioangiol
50:419-29 (2002).
Keloids
Related to scars are lesions known as keloids. Keloids arise from sites of previous
trauma. Keloids are a considerable source of morbidity because of continued growth, pruritus,
and physical appearance. Clinically, keloids are distinguished from scars in that keloids
continue to grow over the borders of the original injury. It has been observed that both
sirolimus and tacrolimus (i.e., FK506; an antiproliferative) effectively treat keloids. Kim et
al, Are Keloid Really "Glib-loids": High-Level Expression Of Glib-1 Oncogeny In Keloid. J.
Am Acad Dermatol 45(5):707-711 (2001).
TNF-B Lieand
Also, a combination of a sirolimus derivative with a tissue growth factor-beta ligand is
known to prevent the formation of ocular scar tissue and/or promote the proliferation of
connective tissue or soft tissue for wound healing. Donahoe et al, Methods And
Compositions For Enhancing Cellular Response To [email protected] Ligands. United States Patent No.
5,912,224.
Clearly, still lacking in the art is any contemplation that sirolimus and analogs of
sirolimus may be effective either during, or after, a surgical procedure to reduce or prevent
the formation of scar tissue on any living tissue.
Preferred Embodiments Of the Present Invention
Excess scar tissue production is a known morbidity consequence of healing from a
number of types of wounds. Examples include, but are not limited to, hypertrophic burn
scars, surgical adhesions (i.e., for example, abdominal, vascular, spinal, neurological, thoracic
and cardiac), capsular contracture following breast implant surgery and excess scarring
following eye surgery and ear surgery.
The delivery of specific compounds contemplated by this invention to a surgical site or
wound include, but are not limited to, microparticles, gels, hydrogels, foams, bioadhesives,

liquids, xerogels or surgical dressings. Particularly, these media are produced in various
embodiments providing a controlled release of a compound such as sirolimus.
CLINICAL APPLICATIONS
Burns
Burn injuries are well known for the development of scar tissue during the healing
process. Sirolimus and analogs of sirolimus are contemplated by the present invention to be
applied by any one of the compositions and methods described herein to facilitate the healing
and reduction and/or prevention of scar tissue and adhesions of a bum wound.
The clinical management of burn-induced hypertrophic scarring has focused primarily
on the application of pressure since the early 1970s. Although the exact mechanism of action
is unknown, pressure appears clinically to enhance the scar maturation process. Bandages that
can be wrapped and unwrapped or are made of a soft material are used in early scar
management. Custom-made pressure garments generally are used for definitive scar
management and inserts are placed in concavities to aid in compression. Staley et al, Use Of
Pressure To Treat Hypertrophic Burn Scars. Adv Wound Care 10:44-46 (1997). However, it
is clear that these approaches, while helpful, still allow the development of serious and
debilitating scarring.
The further development of effective topical chemotherapy, reintroduction of burn
wound excision, and the use of biologic dressings have significantly decreased the incidence
of invasive burn wound infection and have contributed to the improvement in the survival
over the past four decades. The currently available skin substitutes, however, are imperfect
and research endeavors are essential to continue to develop a nonantigenic and disease-free
physiologically effective tissue (i.e., synthetic skin). This approach will eventually improve
wound closure, reduce scar formation thereby reducing the need for reconstructive surgery.
Greenfield et al, Advances In Burn Wound Care. Crit Care Nurs Clin North Am 8:203-15
(1996).
The advent of specific antiproliferative drugs (i.e., sirolimus and analogs of sirolimus)
that reduce scarring in bum patients will provide an enormous benefit to bum patients.

Specifically, by controlling the overgrowth of scar tissue, the normal healing process will be
allowed to predominate. As such, the need for post-burn healing medical treatments to
provide cosmetic, and clinical, treatment for burn scars will be minimized. The present
invention specifically contemplates a method to reduce scars comprising: a) providing;
i) sirolimus or an analog of sirolimus or other cytostatic antiproliferative, ii) a burn patient;
and b) administering said sirolimus or analog of sirolimus to said burn patient under
conditions such that scarring is reduced. Preferably, said sirolimus, analog of sirolimus or
other active compound is delivered locally at a burn site on the skin, either with or without a
systemic concurrent administration of said active compound, such as sirolimus.
Pericarditis
Pericarditis is an inflammation and swelling of the pericardium (/.&, the sac-like
covering of the heart), which can occur in the days or weeks following a heart attack.
Examples of clinical conditions involving pericarditis include, but are not limited to,
Dressler's syndrome, post-myocardial infarction, post-cardiac injury, and postcardiotomy.
Pericarditis may occur within 2 to 5 days after a heart attack (i.e., for example, an
acute myocardial infarction), or it may occur as much as 11 weeks subsequent to such an
attack and may involve repeated episodes of the symptoms. Pericarditis may also result from
open heart surgery, stab wounds to the heart and blunt chest trauma.
Pericarditis occurring shortly after a heart attack is caused by the inflammatory
response to blood in the pericardial sac or by the presence of dead or severely damaged tissue
in the heart muscle. During the period, of inflammation, the immune system sometimes
healthy cells by mistake. Pain occurs when the inflamed pericardium rubs on the heart.
Early pericarditis complicates 7% to 10% of heart attacks. Dressler's syndrome is
seen in only 1% of patients after heart attack. Risks include previous heart attack, open heart
surgery or chest trauma.
In one embodiment, the present invention contemplates a method to reduce scars and
inflammation following heart surgery wherein sirolimus, tacrolimus, analogs of sirolimus or
another cytostatic antiproliferative drug are administered to a patient exhibiting symptoms of
pericarditis. In another embodiment, the present invention contemplates a method to reduce
scars and inflammation wherein sirolimus, tacrolimus, analogs of sirolimus or another

cytostatic antiproliferative drug are administered to a patient undergoing heart surgery so as to
prevent pericarditis.
Surgical Adhesions
Postsurgical adhesions are fibrous scar tissue formations, or fibrin matrices, that form
between tissues or organs following injury associated with surgical procedures. Such injuries
include ischemia, foreign body reaction, hemorrhage, abrasion, incision, and infection-related
inflammation. In the U.S., the annual cost of removing lower abdominal adhesions is
estimated to be more than $2 billion in inpatient treatment charges. Adhesions also develop
following cardiac, spinal, neurological, pleural and other thoracic surgery. In one
embodiment, the present invention contemplates a reduction in pleural adhesions following
lung surgery. In another embodiment, the present invention contemplates a reduction in
cardiac adhesions following cardiac surgery.
Postsurgical damage sites form adhesions in tissues or organs that normally remain
separate, but instead, join together by fibrin matrices within the first few days following
surgery. Under normal circumstances, most fibrin matrices between organs degrade during .
the healing process. When fibrin matrices fail to degrade, permanent adhesions are formed,
linking tissues and/or organs together. Such unwanted adhesion formation following
gynecologic or general abdominal surgeries can lead to a variety of complications, including
pain, infertility and bowel obstruction.
Adhesions are recognized as serious sequelae in patients undergoing gynecologic and
general abdominal surgical procedures. For example, the presence of adhesions between
structures such as the fallopian tubes, ovaries and uterus following surgery is a major cause of
pain and infertility.
Abdominal adhesions are the predominant cause of small-bowel obstruction,
accounting for 54% to 74% of cases. Moreover, approximately 80% to 90% of abdominal
adhesions result from surgery.
Pelvic adhesions occur in 55% to 100% of fertility-enhancing procedures as
determined by second-look laparoscopy performed in a number of large, multicenter studies.
In an attempt to reduce the tissue trauma and thus recovery time, special microsurgical
medical procedures have been developed that minimize tissue handling. However, even when

these techniques are followed, postoperative adhesions can occur in the majority of patients in
certain surgical procedures. Therefore, it is generally believed that the best approach to
minimizing postsurgical adhesion formation is through the use of special microsurgical
techniques in combination with anti-adhesion protocols.
The reduction of post-surgical adhesions following liquid spray applications of
fiuorocarbons to open surgical sites is known. These fluorocarbons act by coating the tissue
and reducing surface tension, thus preventing adherence of the coated tissues when brought
into close proximity. Niazi, S., Use Of Fluorocarbons For The Prevention Of Surgical
Adhesions. United States Patent No. 6,235,796.
Another method to reduce surgical tissue adhesion by physical barrier means utilizes a
dual chamber spray can or bottle that mixes two polymer solutions at the nozzle. This mixing
initiates a nucleophilic-electrophilic crosslinking reaction and generates a solidified polymer
matrix. Either polymer mixture is also capable of delivering growth factors to a surgical site
as part of a bioadhesive polymer matrix. The polymer matrix prevents post-surgical adhesion
formation via the tissue surface coating and is capable of removing scar tissue. Synthetic
polymers of collagen or hyaluronic acid are specifically contemplated, and natural proteins
may be added to improve the bioadhesive properties of the matrix. Rhee et al, Method Of
Using Crosslinked Polymer Compositions In Tissue Treatment Applications. United States
Patent No. 6,116,139 (herein incorporated by reference).
Alternatively, it is known that post-surgical adhesions are prevented or reduced by the
administration of another type of barrier, a thermally gelling polymer. Gelation of a thermal
gel during its administration is determined by its phase transition temperature. It is known in
the art that the thermal gel phase transition temperature may be modified by mixing a
modifier polymer (i.e., cellulose esters or Carbopol®) with a constitutive polymer (i.e.,
polyoxyalkene copolymer). Flore et al, Methods And Compositions For The Delivery Of
Pharmaceutical Agents And/Or The Prevention Of Adhesions. United States Patent No.
6,280,745 (herein incorporated by reference). The '745 patent explains that prevention of
post-surgical scarring and adhesions is due the actual physical presence of the gel (i.e., acting
as an artificial barrier to growth), rather than due to the pharmacological action of any

compound delivered with the hydrogel.
The present invention contemplates the administration of a medium comprising a
cytostatic and antiproliferative compound (i.e., such as, for example, sirolimus, tacrolimus
and/or analogs of sirolimus) to a surgical site or other area of tissue injury that, by
pharmacological action, prevents or reduces the formation of scar tissue and post-surgical
adhesions. In a preferred embodiment, the medium comprising the cytostatic or
antiproliferative compound is easily administered to the surgical field via liquid administration
techniques, via a thermally gelling polymer, via a bioadhesive or via microparticles.
External Vascular Scarring
The present invention contemplates a medium comprising a cytostatic and
antiproliferative compound (i.e., sirolimus, tacrolimus and analogs of sirolimus) applied to an
external vascular site. In one embodiment, the compound reduces or prevents the formation
of scar tissue or tissue adhesions.
The advent of permanent hemodialysis access has made possible the use of chronic
hemodialysis in patients with end-stage renal disease. Although autogenous arteriovenous
fistulae remain the conduit of choice, their construction is not always feasible. Prosthetic
grafts made of polytetrafluoroethylene (PTFE) are typically the second-line choice for
hemoaccess. However, these grafts suffer from decreased rates of patency and an increased
number of complications. Anderson et al., Polytetrafluoroethylene Hemoaccess Site
Infections, American Society for Artificial Internal Organs Journal, 46(6):S 18-21 (2000). In
one embodiment, the present invention contemplates the administration of a medium
comprising sirolimus, tacrolimus or an analog of sirolimus to a patient having PTFE graft
complication. In one emboidment, the medium is sprayed onto the PTFE graft. In another
embodiment, the medium is attached to a surgical wrap that encircles the PTFE graft. In one
embodiment, the medium is attached to a surgical sleeve (i.e., a bandage or mesh that is
tubular in nature) that is placed onto the exterior surface of the vasculature during the PTFE
graft procedure.
Ear Scarring
One aspect of the present invention contemplates a method of applying sirolimus,

tacrolimus and analogs of sirolimus in and around the ear to prevent progressive inner ear
deterioration {i.e., cholesteatoma). It is known that process of scar formation within the ear,
including the ear drum, is very similar to other tissues. The epithelial pathogenesis of
acquired cholesteatoma appears to have three prerequisites: (1) the unique anatomical situation
at the ear-drum (two different epithelial layers close together); (2) chronic destruction of the
submucosal tissue in the middle ear (infection, inflammation); and (3) wound healing (i.e., a
proliferation phase). Destruction of the submucosal space by middle ear infection and cell
necrosis starts the wound healing cascade. In wound healing, generally the connective tissue
fibroblasts and macrophages play a pivotal role. Cytokines are thought to promote the
re-epithelization of the mucosal defect and scar tissue development act upon the intact
squamous cell layer of the outer surface of the ear-drum at the same time. Thereby a
proliferation of the undamaged epithelial layer is induced. Cholesteatoma matrix is always
surrounded by a layer of connective tissue, the perimatrix. Persistence of the inflammation
causes permanent wound healing in the perimatrix, proliferation of the fibroblasts (granulation
tissue) and proliferation of the epithelium (matrix). It is speculated that by virtue of wound
healing cytokines of fibroblasts and macrophages are the driving forces of cholesteatoma
origin, growth and bone destruction. Milewski C, Role Of Perimatrix Fibroblasts In
Development Of Acquired Middle Ear Cholesteatoma. A Hypothesis. HNO 46:494-501
(1998).
The present invention contemplates the administration of a medium comprising a
cytostatic and antiproliferative compound including, but not limited to, sirolimus, tacrolimus
and/or analogs of sirolimus, to the ear so that, by pharmacological action, such excess scar
tissue is prevented or reduced.
Eve Scarrine
One aspect of the present invention contemplates a method of applying sirolimus,
tacrolimus, and analogs of sirolimus to eye tissues following or during surgery or trauma.
Various conditions of the eye are known to be associated with corneal scarring and fibroblast
proliferation, including ocular coagulation and burns, mechanical and chemical injury, ocular
infections such as kerato-conjunctivitis, and other ocular conditions. Some of these conditions
are known to arise post-operatively after surgical treatment of other ocular conditions. This

undesirable tissue growth is easily neovascularized and therefore becomes permanently
established and irrigated. Tissue scarring or fibroblast proliferation is a condition which is
difficult to treat. Presently, it is treated by subjecting the ocular area to further surgery or by
using steroids, topically or by injection. However, steroids do increase side effects such as
infection, cataract and glaucoma. Other non-steroidal agents like indomethcin
have very little anti-scarring effects. (Williamson J. et al., British J. of Ophthalmology 53:361
(1969); Babel, J., Histologie Der Crtisonkatarakt, p.327. Bergmann, Munich (1973)).
It is known in the art that corneal scarring, neovascularization or fibroblast
proliferation maybe reduced by the application of a human leukocyte elastase (HLE)
inhibitory agents (i.e., carbamates substituted by oligopeptides). Digenis et al. Methods Of
Treating Eye Conditions With Human Leukocyte Elastase (HLE) Inhibitory Agents. United
States Patent No. 5,922,319. Elastases (human leukocyte elastase and cathepsin G), appear to
be responsible for some chronic tissue destruction associated with inflammation, arthritis and
emphysema. Therefore, the actions of elastase inhibitors do not involve the mTOR protein in
regards to their antiproliferative effects relative to the reduction of scar tissue formation.
Progressive scarring may result in blindness, especially in cases where the retina is
involved. The most common cause of failure of retinal reattachment surgery is formation of
fibrocellular contractile membranes on both surfaces of the neuroretina. This intraocular
fibrosis, known as proliferative vitreoretinopathy, results in a blinding fractional retinal
detachment because of the contractile nature of the membrane. Contractility is a
cell-mediated event that is thought to be dependent on locomotion and adhesion to the
extracellular matrix. Sheridan et al.. Matrix Metalloproteinases: A Role In "Die Contraction
Of Vitreo-Retinal Scar Tissue. Am J Pathol 159:1555-66 (2001).
Corneal wound healing frequently leads to the formation of opaque scar tissue.
Stromal fibroblastic cells of injured corneas express collagen IV and contribute to the
formation of a basal lamina-like structure. Normally, stromal collagenous matrix organizes in
orthogonal lamellae during corneal development, whereas that of an alkali-burned cornea, is
known to develop in a disorganized manner. Enhanced expression of collagen IV by the
fibroblastic cells in the stroma of injured corneas is consistent with the notion that they may

contribute to the formation of basal lamina-like structures in injured corneas. Ishizaki et al,
Stromal Fibroblasts Are Associated With Collagen W In Scar Tissues Of Alkali-Burned And
Lacerated Corneas. Curr Eye Res 16:339-48 (1997).
Medical Devices
One aspect of the present invention contemplates a method for applying sirolimus,
tacrolimus and analogs of sirolimus to reduce scar tissue formation and adhesions following
the placement of medical device implants.
Excess scar tissue formation and inflammation around direct medical implants are of
particular concern. For example, the permanent placement of a percutaneous functional
implant that protrudes through the skin for prolonged periods of time has not yet become a
reality. Efforts towards eventual success must be directed toward a variety of failure
mechanisms. For example, these mechanisms may be either extrinsic or intrinsic that cause
shearing and tearing at the skin-implant interface. Extrinsic forces are defined as those forces
applied either to the skin or the implant by the external environment. Intrinsic forces are
those that have to do directly or indirectly with the body's growth and cell maturation, such
as the retraction of maturing scar tissue and the surface migration of squamous epithelium.
An intact skin-implant interface is important to attain in order to provide a seal against
microbial invasion. The skin must remain intact, since a suppurative wound makes the
implant's removal mandatory. Hall et al, Some Factors That Influence Prolonged Interfacial
Continuity. J Biomed Mater Res 18:383-93 (1984).
Implants for reconstructive or cosmetic surgery, such as breast implants, also have
problems with excess scar tissue formation. Breast implants are known to develop
surrounding scar capsules that may harden and contract, resulting in discomfort, weakening of
the shell with rupture, asymmetry, and patient dissatisfaction. This phenomenon is known to
occur in as many as 70 percent of implanted patients over time. Most complications are due
to late leaks, infection, and capsular contracture. Ersek et al, Textured Surface, Nonsilicone
Gel Breast Implants: Four Years' Clinical Outcome. Plast Reconstr Surg 100:1729-39
(1997).
Glaucoma implants are also suspected to fail due to scar formation. Glaucoma

implants are designed to increase fluid outflow from the eye in order to decrease intraocular
pressure and prevent damage to the optic nerve. The implant consists of a silicone tube that
is inserted into the anterior chamber at one end and is attached at the other end to a silicone
plate that is sutured to the outside of the globe beneath the conjunctiva. The glaucoma
"implant" becomes a "drain" over the first 3 to 6 postoperative weeks as the silicone plate is
enclosed by a fibrous capsule that allows a space to form into which fluid can drain and from
which fluid can be absorbed by the surrounding tissues. Ideally, the size and thickness of the
capsule (i.e., the filtering bleb) that surrounds the plate is such that the amount of fluid that
passes through the capsule is identical to the amount of fluid produced by the eye at an
intraocular pressure of 8 to 14 mmHg. The most common long-term complication of these
implants is failure of the filtering bleb 2 to 4 years after surgery due to the formation of a
thick fibrous capsule around the device. Micromovement of the smooth drainage plate against
the scleral surface may be integral to the mechanism of glaucoma implant failure by
stimulating low-level activation of the wound healing response, increased collagen scar
formation, and increased fibrous capsule thickness. Jacob et al. Biocompatibility Response To
Modified Baerveldt Glaucoma Drains. J Biomed Mater Res 43:99-107 (1998).
Another aspect of the present invention contemplates coating a medical device with a
medium comprising sirolimus, tacrolimus or an analog of sirolimus. A "coating", as used
herein, refers to any compound that is attached to a medical device. For example, such
attachment includes, but is not limited to, surface adsorption, impregnation into the material
of manufacture, covalent or ionic bonding and simple friction adherence to the surface of the
medical device.
Sirolimus or analogs of sirolimus may be attached to a medical device in a number of
ways and utilizing any number of biocompatible materials (i.e., polymers). Different
polymers containing sirolimus are utilized for different medical devices. For example, a
ethylene-co-vinylacetate and polybutylmethacrylate polymer is utilized with stainless steel.
Falotico et al, United States Patent Application, 20020016625. Other polymers may be
utilized more effectively with medical devices formed from other materials, including
materials that exhibit superelastic properties such as alloys of nickel and titanium. In one

embodiment, a compound such as, but not limited to, sirolimus, tacrolimus or analogs of
sirolimus are directly incorporated into a polymeric matrix and sprayed onto the outer surface
of a catheter such that the polymeric spray becomes attached to said catheter. In another
embodiment, said compound will then elute from the polymeric matrix over time and enter
the surrounding tissue. In one embodiment, said compound is expected to remain attached on
the catheter for at least one day up to approximately six months.
In one embodiment, the present invention contemplates a sirolimus hydrogel polymer
coating on a stainless steel medical device (i.e., for example, a permanent implant)..
Preferably, a stainless steel implant is brush coated with a styrene acrylic aqueous dispersion
polymer (55% solids) and dried for 30 minutes at 85°C. Next, this polymer surface is
overcoated with a controlled release hydrogel composition consisting of:

The coating is then dried for 25 hours at 85°C prior to use. It is not intended that the present
invention be limited by the above sirolimus concentration. One skilled in the art should
realize that that various concentrations of sirolimus may be used such as, but not limited to,
1.0 - 10 mg/ml, preferably 0.1 - 5 mg/ml, and more preferably 0.001 - 1 mg/ml.
In another embodiment, a multiple layering of non-erodible polymers may be utilized
in conjunction with sirolimus. Preferably, the polymeric matrix comprises two layers; a inner
base layer comprising a first polymer and the incorporated sirolimus and an outer second
polymer layer acting as a diffusion barrier to prevent the sirolimus from eluting too quickly
and entering the surrounding tissues. In one embodiment, the thickness of the outer layer or
top coat determines the rate at which the sirolimus elutes from the matrix. Preferably, the
total thickness of the polymeric matrix is in the range from about 1 micron to about 20
microns or greater. Another embodiment of the present invention contemplates spraying or

dipping a polymer/sirolimus mixture onto a catheter.
Intraluminal Narrowing
The formation of excess scar tissue and resultant intraluminal narrowing in bodily
lumens is a well known phenomenon following illness, injurious trauma, implants or surgery
that involves bodily organs. The mechanisms for such narrowing include fibroblastic,
endothelial and intimal excess proliferation or hyperplasia. Perhaps the most well-known
condition is that of restenosis, which is a condition of a narrowing of the vascular lumen
following systemic or local hyperproliferative vascular disease, or as a complication of
vascular surgery, injurious trauma or implantation of a medical device. Other examples of
excess luminal narrowing occur following ductal/tubal surgery, including, but not limited to,
pancreatic, biliary and fallopian tube surgery.
Although it is not intended to limit the present invention, it is believed that the
following example regarding arteriovenous fistula blockage provides an adequate teaching.
Vascular access complications include, but are not limited to, arteriovenous fistulae
which is a major problem in hemodialysis patients. The most common complication is
progressive stenosis at the anastomotic site. In most cases, this stenosis occurs at the venous
anastomotic site.
Vascular access is governed by the DOQI (Dialysis Outcome Quality Initiatives). In
early 2000, the National Kidney Foundation (NKF) announced it is expanding the scope of
DOQI study to include "all phases of kidney disease and dysfunction and their monitoring and
management." DOQI has developed and published clinical practice guidelines in four areas -
hemodialysis, peritoneal dialysis, anemia, vascular access and nutrition.
Thus, according to DOQI, patients requiring vascular access are treated with the
following progression of dialysis vascular access grafts as they fail: i) a Cimino graft; which
is a lower forearm radial artery/cephalic vein A-V fistula (i.e., a native graft); ii) an upper
arm native fistula; connecting the brachial artery to either the cephalic or basilic vein; and iii)
an upper arm PTFE Loop; connecting the brachial artery to the median antecubical vein.
The primary failure issues related to graft technology are that: i) even though 70%
Cimino grafts are suitable for use 50% fail over the first ten years and 30% generate
thromboses or fail to mature (i.e., undergo endothelization and healing); ii) a condition

known as "steal" develops that is characterized by a high blood flow rate through the graft
(i.e., 300- 500 ml/min) resulting in a lack of blood flow to the hand and lower arm and iii)
PTFE grafts typically develop initimal thickening at the venous anatomotic site.
One approach to remedy these problems is to apply a perivascular endothelial cell
implant to inhibit intimal thickening observed following chronic arteriovenous anastomoses.
Nugent et al, Perivascular Endothelial Implants Inhibit Intimal Hyperplasia In A Model Of
Arteriovenous Fistulae: A Safety And Efficacy Study In The Pig. J Vase Res 39(6):524-33
(2002). In one embodiment, the present invention contemplates a method to reduce scar
tissue formation following an arteriovenous anastomosis in a dialysis patient. In another
embodiment, said patient has end stage renal disease. In another embodiment, the patient has
an artificial graft.
Another aspect of the present invention contemplates treatment of vascular
complications following coronary or peripheral bypass graft surgeries. It is well known that
arterial grafts have a higher success rate than autologous venous grafts. However, venous
grafts remain preferred as they are easier to harvest and insert and far more available. A
major disadvantage to using venous grafts lies in the fact that 10%-18% fail within 1-6
months following surgery, due predominately to exaggerated intimal hyperplasia. Hyperplasia
may be accompanied by neointimal thickening and atherosclerotic plaques. Improvement in
vein graft patency, therefore, remains a long felt need in this area of vascular surgery.
The present invention contemplates a method to improve the patency of vascular grafts
by administration of a medium comprising sirolimus, tacrolimus and analogs of sirolimus
following any surgical manipulation (i.e, for example, suturing) that results in a direct trauma
to the endothelium and smooth muscle cells of the vasculature. In one embodiment, the
administration of said medium reduces anastomotic and vein graft intimal hyperplasia believed
caused by an intrinsic adaptive response of the medial smooth muscle cells.
Transplantations
One aspect of the present invention contemplates a medium comprising a cytostatic
and antiproliferative compound {i.e., for example, sirolimus, tacrolimus and analogs of

sirolimus) administered to a patient during and after an organ transplant. In one embodiment,
a method results in the prevention or reduction of post-transplantation scarring. It is well
known in the art that sirolimus and related compounds are effective in reducing the graft-
versus-host rejection cascade. This invention, however, proposes a novel use in regards to
prevention of scarring for sirolimus in this clinical setting.
DRUG DELIVERY SYSTEMS
The present invention contemplates several drug delivery systems that provide for
roughly uniform distribution, have controllable rates of release and may be administered to
either an open or closed surgical site. A variety of different media are described below that
are useful in creating drug delivery systems. It is not intended that any one medium or carrier
is limiting to the present invention. Note that any medium or carrier may be combined with
another medium or carrier; for example, in one embodiment a polymer microparticle carrier
attached to a compound may be combined with a gel medium.
Carriers or mediums contemplated by this invention comprise a material selected from
the group comprising gelatin, collagen, cellulose esters, dextran sulfate, pentosan polysulfate,
chitin, saccharides, albumin, fibrin sealants, synthetic polyvinyl pyrrolidone, polyethylene
oxide, polypropylene oxide, block polymers of polyethylene oxide and polypropylene oxide,
polyethylene glycol, acrylates, acrylamides, methacrylates including, but not limited to, 2-
hydroxyethyl methacrylate, poly(ortho esters), cyanoacrylates, gelatin-resorcin-aldehyde type
bioadhesives, polyacrylic acid and copolymers and block copolymers thereof.
One aspect of the present invention contemplates a medical device comprising several
components including, but not limited to, a reservoir comprising sirolimus, tacrolimus or an
analog of sirolimus, a catheter, a sprayer or a tube. In one embodiment, said medical device
administers either an internal or external spray to a patient. In another embodiment, said
medical device administers either an internal or external gel to a patient.
One embodiment of the present invention contemplates a drug delivery system
comprising sirolimus, tacrolimus (FK506) and analogs of sirolimus such as, but not limited to,
everolimus (i.e., SDZ-RAD), CCT-779, ABT-578, 7-epi-rapamycin, 7-thiomethyl-rapamycin,
7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin, 7-demethoxy-rapamycin, 32-
.48.

demethoxy-rapamycin and 2-desmethyl-rapamycin.
Other derivatives of sirolimus comprising mono-esters and di-esters at
positions 31 and 42 have been shown to be useful as antifungal agents (U.S. Pat. No.
4,316,885) and as water soluble prodrugs of rapamycin (U.S. Pat. No. 4,650,803). A
30-demethoxy rapamycin has also been described in the literature (C. Vezina et al. J. Antibiot.
(Tokyo), 1975, 28 (10), 721; S. N. Sehgal et al., J. Antibiot. (Tokyo), 1975, 28(10), 727;
1983, 36(4), 351; N. L. Pavia et al., J. Natural Products, 1991, 54(1), 167-177).
Numerous other chemical modifications of rapamycin have been attempted. These
include the preparation of mono- and di-ester derivatives of rapamycin (WO 92/05179),
27-oximes of rapamycin (EP0 467606); 42-oxo. Analog of rapamycin (U.S. Pat. No.
5,023,262); bicyclic rapamycins (U.S. Pat. No. 5,120,725); rapamycin dimers (U.S. Pat. No.
5,120,727); silyl ethers of rapamycin (U.S. Pat. No. 5,120,842); and arylsulfonates and
sulfamates (U.S. Pat. No. 5,177,203). Rapamycin was recently synthesized in its naturally
occurring enantiomeric form (K. C. Nicolaou et al., J. Am. Chem. Soc, 1993, 115,
4419-4420; S. L. Schreiber, J. Am. Chem. Soc, 1993, 115, 7906-7907; S. J. Danishefsky, J.
Am. Chem. Soc, 1993, 115, 9345-9346.
Alternatively, media may also comprise non-sirolimus compounds, such as, but not
limited to, antisense c-myc and tumstatin. Other pharmaceutical compounds may be delivered
either alone or in combination with sirolimus and analogs of sirolimus, such as, but not
limited to, antiinflammatory, corticosteroid, antithrombotic, antibiotic, antifungal, antiviral,
analgesic and anesthetic. .
Microparticles
One aspect of the present invention contemplates a medium comprising a
microparticle. Preferably, microparticles comprise liposomes, nanoparticles, microspheres,
nanospheres, microcapsules, and nanocapsules. Preferably, some microparticles contemplated
by the present invention comprise poly(lactide-co-glycolide), aliphatic polyesters including,
but not limited to, poly-glycolic acid and poly-lactic acid, hyaluronic acid, modified
polysacchrides, chitosan, cellulose, dextran, polyurethanes, polyacrylic acids, psuedo-
poly(amino acids), polyhydroxybutrate-related copolymers, polyanhydrides,
polymethylmethacrylate, poly(ethylene oxide), lecithin and phospholipids.

Liposomes
One aspect of the present invention contemplates liposomes capable of attaching and
releasing sirolimus and analogs of sirolimus. Liposomes are microscopic spherical lipid
bilayers surrounding an aqueous core that are made from amphiphilic molecules such as
phospholipids. For example, Figure 1 demonstrates one liposome embodiment where a
sirolimus molecule 2 is trapped between hydrophobic tails 4 of the phospholipid micelle 8.
Water soluble drugs can be entrapped in the core and lipid-soluble drugs, such as sirolimus,
can be dissolved in the shell-like bilayer. Lipesomes have a special characteristic in that they
enable water soluble and water insoluble chemicals to be used together in a medium without
the use of surfactants or other emulsifiers. As is well known in the art, liposomes form
spontaneously by forcefully mixing phosopholipids in aqueous media. Water soluble
compounds are dissolved in an aqueous solution capable of hydrating phospholipids. Upon
formation of the liposomes, therefore, these compounds are trapped within the aqueous
liposomal center. The liposome wall, being a phospholipid membrane, holds fat soluble
materials such as oils. Liposomes provide controlled release of incorporated compounds. In
addition, liposomes can be coated with water soluble polymers, such as polyethylene glycol to
increase the pharmacokinetic half-life. One embodiment of the present invention contemplates
an ultra high-shear technology to refine liposome production, resulting in stable, unilamellar
(single layer) liposomes having specifically designed structural characteristics. These unique
properties of liposomes, allow the simultaneous storage of normally immiscible compounds
and the capability of their controlled release.
The present invention contemplates cationic and anionic liposomes, as well as
liposomes having neutral lipids comprising sirolimus and analogs of sirolimus. Preferably,
cationic liposomes comprise negatively-charged materials by mixing the materials and fatty
acid liposomal components and allowing them to charge-associate. Clearly, the choice of a
cationic or anionic liposome depends upon the desired pH of the final liposome mixture.
Examples of cationic liposomes include lipofectin, lipofectamine, and lipofectace.
One embodiment of the present invention contemplates a medium comprising
liposomes that provide controlled release of sirolimus and analogs of sirolimus. Preferably,
liposomes that are capable of controlled release: i) are biodegradable and non-toxic; ii) carry

both water and oil soluble compounds; iii) solubilize recalcitrant compounds; iv) prevent
compound oxidation; v) promote protein stabilization; vi) control hydration; vii) control
compound release by variations in bilayer composition such as, but not limited to, fatty acid
chain length, fatty acid lipid composition, relative amounts of saturated and unsaturated fatty
acids, and physical configuration; viii) have solvent dependency; iv) have pH-dependency and
v) have temperature dependency.
The compositions of liposomes are broadly categorized into two classifications.
Conventional liposomes are generally mixtures of stabilized natural lecithin (PC) that may
comprise synthetic identical-chain phospholipids that may or may not contain glycolipids.
Special liposomes may comprise: i) bipolar fatty acids; ii) the ability to attach antibodies for
tissue-targeted therapies; iii) coated with materials such as, but not limited to lipoprotein and
carbohydrate; iv) multiple encapsulation and v) emulsion compatibility.
Liposomes may be easily made in the laboratory by methods such as, but not limited
to, sonication and vibration. Alternatively, compound-delivery liposomes are commercially
available. For example, Collaborative Laboratories, Inc. are known to manufacture custom
designed liposomes for specific delivery requirements.
Microspheres, Microparticles And Microcapsules
Microspheres and microcapsules are useful due to their ability to maintain a generally
uniform distribution, provide stable controlled compound release and are economical to
produce and dispense. Preferably, an associated delivery gel or the compound-impregnated
gel is clear or, alternatively, said gel is colored for easy visualization by medical personnel.
One of skill in the art should recognize that the terms "microspheres, microcapsules and
microparticles" (i.e., measured in terms of micrometers) are synonymous with their respective
counterparts "nanospheres, nanocapsules and nanoparticles" {i.e., measured in terms of
nanometers). It is also clear that the art uses the terms "micro/nanosphere, micro/nanocapsule
and micro/nanoparticle" interchangeably, as will the discussion herein.
Microspheres are obtainable commercially (Prolease®, Alkerme's: Cambridge, Mass.).
For example, a freeze dried sirolimus medium is homogenized in a suitable solvent and
sprayed to manufacture microspheres in the range of 20 to 90 μm. Techniques are then
followed that maintain sustained release integrity during phases of purification, encapsulation

and storage. Scott et al, Improving Protein Therapeutics With Sustained Release
Formulations, Nature Biotechnology, Volume 16:153-157 (1998).
Modification of the microsphere composition by the use of biodegradable polymers can
provide an ability to control the rate of sirolimus release. Miller et al., Degradation Rates of
Oral Resorbable Implants {Polylactates and Polyglycolates: Rate Modification and Changes
in PLAJPGA Copolymer Ratios, J. Biomed. Mater. Res., Vol. 11:711-719 (1977).
Alternatively, a sustained or controlled release microsphere preparation is prepared
using an in-water drying method, where an organic solvent solution of a biodegradable
polymer metal salt is first prepared. Subsequently, a dissolved or dispersed medium of
sirolimus is added to the biodegradable polymer metal salt solution. The weight ratio of
sirolimus to the biodegradable polymer metal salt may for example be about 1:100000 to
about 1:1, preferably about 1:20000 to about 1:500 and more preferably about 1:10000 to
about 1:500. Next, the organic solvent solution containing the biodegradable polymer metal
salt and sirolimus is poured into an aqueous phase to prepare an oil/water emulsion. The
solvent in the oil phase is then evaporated off to provide microspheres. Finally, these
microspheres are then recovered, washed and lyophilized. Thereafter, the microspheres may
be heated under reduced pressure to remove the residual water and organic solvent.
Other methods useful in producing microspheres that are compatible with a
biodegradable polymer metal salt and sirolimus mixture are: i) phase separation during a
gradual addition of a coacervating agent; ii) an uvwater drying method or phase separation
method, where an antiflocculant is added to prevent particle agglomeration and iii) by a
spray-drying method.
In one aspect the present invention contemplates a medium comprising a microsphere
or microcapsule capable of delivering a controlled release of a compound for a duration of
approximately between 1 day and 6 months. In one embodiment, the microsphere or
microparticle may be colored to allow the medical practitioner the ability to see the medium
clearly as it is dispensed. In another embodiment, the microsphere or microcapsule may be
clear. In another embodiment, the microsphere or microparticle is impregnated with a radio-
opaque fluoroscopic dye.
Controlled release microcapsules may be produced by using known encapsulation

techniques such as centrifugal extrusion, pan coating and air suspension. Using techniques
well known in the state of the art, these microspheres/microcapsules can be engineered to
achieve particular release rates. For example, Oliosphere® (Macromed) is a controlled release
microsphere system. These particular microsphere's are available in uniform sizes ranging
between 5 - 500 μm and composed of biocompatible and biodegradable polymers. It is well
known in the art that specific polymer compositions of a microsphere control the drug release
rate such that custom-designed microspheres are possible, including effective management of
the burst effect. ProMaxx® (Epic Therapeutics, Inc.) is a protein-matrix drug delivery system.
The system is aqueous in nature and is adaptable to standard pharmaceutical drug delivery
models. In particular, ProMaxx® are bioerodible protein microspheres that deliver both small
and macromolecular drugs, and may be customized regarding both microsphere size and
desired drug release characteristics.
In one embodiment, a microsphere or microparticle comprises a pH sensitive
encapsulation material that is stable at a pH less than the pH of the internal mesentery. The
typical range in the internal mesentery is pH 7.6 to pH 7.2. Consequently, the microcapsules
should be maintained at a pH of less than 7. However, if pH variability is expected, the pH
sensitive material can be selected based on the different pH criteria needed for the dissolution
of the microcapsules. The encapsulated compound, therefore, will be selected for the pH
environment in which dissolution is desired and stored in a pH preselected to maintain
stability. Examples of pH sensitive material useful as encapsulants are Eudragit® L-100 or
S-100 (Rohm GMBH), hydroxypropyl methylcellulose phthalate, hydroxypropyl
methylcellulose acetate succinate, polyvinyl acetate phthalate, cellulose acetate phthalate, and
cellulose acetate trimellitate. In one embodiment, lipids comprise the inner coating of the
microcapsules. In these compositions, these lipids may be, but are not limited to, partial
esters of fatty acids and hexitiol anhydrides, and edible fats such as triglycerides. Lew C. W.,
Controlled-Release pH Sensitive Capsule And Adhesive System And Method. United States
Patent No. 5,364,634 (herein incorporated by reference).
One embodiment of the present invention contemplates microspheres or microcapsules
comprising sirolimus, tacrolimus (FK506) and analogs of sirolimus such as, but not limited to,
everolimus (i.e., SDZ-RAD), CCI-779, ABT-578, 7-epi-sirolimus, 7-thiomethyl-sirolimus,

7-epi-trimethoxyphenyl-sirolimus, 7-epi-thiomethyl- sirolimus, 7-demethoxy- sirolimus,
32-demethoxy-sirolimus and 2-desmethyl-sirolimus. Alternatively, microspheres or
microcapsules may also comprise non-sirolimus compounds such as, but not limited to,
antisense to c-myc and tumstatin. Other, complementary pharmaceutical compounds may be
delivered either alone or in combination with sirolimus and analogs of sirolimus, such as, but
not limited to, antiinflammatory, corticosteriod, antithrombotic, antibiotic, antifungal, antiviral,
analgesic and anesthetic.
In one embodiment, a microparticle contemplated by this invention comprises a
gelatin, or other polymeric cation having a similar charge density to gelatin (i.e., poly-L-
lysine) and is used as a complex to form a primary microparticle. A primary microparticle is
produced as a mixture of the following composition: i) Gelatin (60 bloom, type A from
porcine skin), ii) chondroitin 4-sulfate (0.005% - 0.1%), iii) glutaraldehyde (25%, grade 1),
and iv) l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride
(EDC hydrochloride), and ultra-pure sucrose (Sigma Chemical Co., St. Louis, Mo.). The
source of gelatin is not thought to be critical; it can be from bovine, porcine, human, or other
animal source. Typically, the polymeric cation is between 19,000-30,000 daltons.
Chondroitin sulfate is then added to the complex with sodium sulfate, or ethanol as a
coacervation agent.
Following the formation of a microparticle, a compound (i.e., for example, sirolimus)
is directly bound to the surface of the microparticle or is indirectly attached using a "bridge"
or "spacer". The amino groups of the gelatin lysine groups are easily derivatized to provide
sites for direct coupling of a compound. Alternatively, spacers (i.e., linking molecules and
derivatizing moieties on targeting ligands) such as avidin-biotin are also useful to indirectly
couple targeting ligands to the microparticles. Stability of the microparticle is controlled by
the amount of glutaraldehyde-spacer crosslinking induced by the EDC hydrochloride. A
controlled release medium is also empirically determined by the final density of
glutaraldehyde-spacer crosslinks.
Table 1 identifies one embodiment for a microcapsule delivery system for sirolimus.
This particular embodiment forms a compound-containing microcapsule bioadhesive gel by
contacting the outer microcapsule surface with an adhesive.


The bioadhesives of this embodiment allow microcapsules to be placed within the
internal mesentery for a sustained period of time for delivery of the compounds contemplated
herein. One skilled in the art should realize that various concentrations of sirolimus may be
incorporated into the above example (i.e., for example, 0.001% - 30%).
In one embodiment, the present invention contemplates microparticles formed by
spray-drying a composition comprising fibrinogen or thrombin with sirolimus and analogs of
sirolimus. Preferably, these microparticles are soluble and the selected protein (i.e.,
fibrinogen or thrombin) creates the walls of the microparticles. Consequently, sirolimus and
analogs of sirolimus are incorporated within, and between, the protein walls of the
microparticle. Heath et al, Microparticles And Their Use In Wound Therapy. United States
Patent No. 6,113,948 (herein incorporated by reference). Following the application of the
microparticles to living tissue, the subsequent reaction between the fibrinogen and thrombin
creates a tissue sealant thereby releasing the incorporated compound into the immediate
surrounding area. In one embodiment, the released compound has.pharmacologic activity
resulting in the reduction of scar tissue formation and/or prevention of tissue adhesion.
In one embodiment (Figure 2), the present invention contemplates a microsphere 10

comprising a biocompatible, biodegradable material into which a cytostatic or antiproliferative
compound (i.e., sirolimus or an analog of sirolimus) 12 is impregnated (i.e., encapsulated).
The compound 12 is contemplated as existing either as fully dissolved or as a colloid.
In one embodiment, (Figure 3), the present invention contemplates a microsphere 10
comprising a biocompatible, biodegradable material into which a cytostatic or antiproliferative
compound (i.e., sirolimus or an analog or sirolimus) 12 is adhered to the microsphere 10
surface.
In another embodiment (Figure 4), the present invention contemplates a microsphere
20 comprising an interior portion 22 comprising a biocompatible, biodegradable material
surrounded by a compound layer 12 of a cytostatic, anti-proliferative compound (i.e.,
sirolimus or an analog of sirolimus) which in turn is surrounded by a second biocompatible,
biodegradable material layer 26. Second layer 26 is capable of controlling the rate of release
of compound layer 12. Preferably, compound layer 12 is released over a time period of
approximately between 1 day and 6 months, hi one specific embodiment, compound layer 12
may be contained within a layer 26 or within the interior portion 22.
The diameter of the exemplary microspheres in either Figure 2 or Figure 3 should be
approximately between 0.1 and 100 microns; preferably 20-75 microns; and more preferably
40-60 microns. One having skill in the art will understand that the shape of the microspheres
need not be exactly spherical; only as very small particles capable of being sprayed or spread
into or onto a surgical site (i.e., either open or closed). In one embodiment, microparticles
are comprised of a biocompatible and/or biodegradable material selected from the group
consisting of polylactide, polyglycolide and copolymers of lactide/glycolide (PLGA),
hyaluronic acid, modified polysaccharides and any other well known material.
The present invention contemplates the combination of microparticles with another
medium described herein. For example, a microparticle may be combined with a medium
including, but not limited to, a foam, hydrogel, gel or a liquid. In one embodiment a
controlled release medium is created. The description of the present invention presents
several exemplary embodiments for a variety of mediums. It is not intended for any
controlled release medium to be limited to combinations described herein.
Liquid Administration

One aspect of the present invention contemplates the administration of a medium
comprising a flowable liquid. Preferably, the liquid media can be administered using a
variety of techniques including, but are not limited to, spraying, pouring, squeezing, and the
like.
In one embodiment, the present invention contemplates a liquid spray medium
comprising liquids, foams, hydrogels, bioadhesives and the like, with or without
microparticles. One embodiment of the present invention contemplates spray mediums
comprising sirolimus, tacrolimus and analogs of sirolimus. Preferably, a spray may be
administered using a catheter directly onto a closed surgical site during an endoscopy
procedure such as, but not limited to, laparoscopy or arthroscopy. Alternatively, a spray of
said compound may generated by a pressure source (i.e., a spray can or a cylinder comprising
a pressure regulator and nozzled tip) to create a droplet spray onto an open surgical site. In
another embodiment, a nebulizer (i.e., for example, an atomizer) may also be used to create
an aerosol spray. In another embodiment, a spray is administered to an open surgical site.
One embodiment of the present invention (Figure 5) contemplates a pressurized spray
can 1 that is capable of spraying a cytostatic anti-proliferative compound (i.e., sirolimus and
analogs of sirolimus) into a surgical or wound site. Pressing an actuator button 3 on top of
the can body 2 causes the compound spray 5 to exit a nozzle 4. Spray 5 is contemplated to
comprise a sirolimus or an analog of sirolimus containing medium selected from the group
consisting of an aqueous mixture, microparticles, foam, and bioadhesive. Alternatively,
nozzel 4 is attached to a medical tubing 6 or a nebulizer (see Figure 6) may also be used to
spray the compound into the surgical site. One having skill in the art would understand that
the present invention is not intended to limit the spraying from a can.
One aspect of the present invention contemplates a method of applying a medium of
sirolimus, tacrolimus and analogs of sirolimus, such as, but not limited to liquids,
bioadhesives and foams to an internal tissue or organ in an even and controlled manner by a
hand-held applicator. In one embodiment, the applicator includes a pump, a tubular extension
that is thin enough to pass through an endoscopic lumen, a proximal end of the tubular
extension being sealingly connected to the pump, and an applicator tip that attaches to the
distal end of the tubular extension. Activation of the pump moves the medium through the tip

and onto the internal tissue in an even and controlled manner without contact of the liquid by
the pump. In one embodiment, the pump is a micropipetter that includes a hand-held portion
having a hand-actuated plunger that does not come in direct, physical contact with the liquid
to be dispensed. The device may further include a wound closure device including at least
two closure pins extending from the distal end of the tubular extension. In another
embodiment, the applicator may be a syringe with a tube extending from the distal end of the
syringe. In another embodiment, the tubular extension is large enough for medical personnel
to firmly grasp by the hands and apply the medium to an open surgical site. In one
embodiment, the applicator comprises two tubular extensions that merge to form a single
applicator tip. Preferably, the two tubular extensions contain different media that are applied
to the tissue as a single mixture. In one embodiment, the tubular extension contains a powder
medium of sirolimus, tacrolimus and analogs of tacrolimus.
In one embodiment, the present invention contemplates a method for spraying a
medium comprising sirolimus and analogs of sirolimus onto an open surgical site. Preferably,
the sprayed medium comprises a bioadhesive requiring the activation of fibrinogen. Gas-
propelled devices are known to spray a first application comprising a first agent capable of
gelling or solidifying and then spraying a second application of a second agent that activates
said first agent to gel or solidify. Epstein G., Gas Driven Spraying Of Mixed Sealant Agents.
United States Patent No. 6,461,361 (herein incorporated by reference). Alternatively, the first
and second agents are mixed during spraying such that they are forming a solid matrix as the
spray contacts the living tissue. Specifically, one type of sterile-gas ejected bioadhesive spray
applicator uses the combination of a protein solution (i.e., thrombin) and a coagulation
solution (i.e., fibrinogen). Fukunaga et ah, Applicator For Applying A Biocompatible
Adhesive. United States Patent No.5,582,596 (herein incorporated by reference).
Alternatively, a metered application of an aerosolized fibrinogen/thrombin bioadhesive is
known by using the step-wise mechanical advancement of two syringes in response to a hand-
held trigger mechanism shaped similarly to a pistol. Coelho et al., Sprayer For Fibrin Glue.
United States Patent No. 5,759,171 (herein incorporated by reference). In another
embodiment, microspheres suspended in a liquid carrier are sprayed. In another embodiment,

a thermally gelling polymer is sprayed into an open surgical site.
In one embodiment, the present invention contemplates a method for spraying a
medium comprising sirolimus and analogs of sirolimus onto a closed surgical site and
surrounding tissues. Preferably, application of liquids to a closed surgical site serves as an
adjunct to the deployment of a sheet of material by an endoscopic surgical device. In one
embodiment, the endoscopic device has multiple openings to dispel a liquid (i.e., saline)
during the deployment of the sheet of material. Tilton et al., Instrumentation For Endoscopic
Surgical Insertion And Application Of Liquid, Gel And Like Material. United States Patent
No. 6,416,506 (herein incorporated by reference). Alternatively, an endoscopic applicator
device (i.e., for example, a spray device adapted for use in a laparoscope) is also
contemplated to selectively direct a spray application of tissue bioadhesives comprising
sirolimus, tacrolimus and analogs of sirolimus. Trumbull, H.R., Laparoscopic Sealant
Applicator. United States Patent No. 6,228,051 (herein incorporated by reference).
Alternatively, a spray tube or device adapted for use via a catheter in an endoscopic or
fluoroscopic device is also contemplated to selectively direct a spray or flow of liquid or gel
media comprising cytostatic pharmaceutical compounds (e.g., sirolimus or analogs thereof) to
a surgical site.
The present invention contemplates laparoscopic devices capable of delivering a
variety of liquid and gel media, including thermoplastic polymers, comprising cytostatic and
antiproliferative drugs (i.e., for example, sirolimus, tacrolimus and analogs of sirolimus),
biologically-active agents and/or water-insoluble thermoplastic polymers to an area of interest
(i.e., for example, an open or closed surgical site). In one embodiment, the invention
contemplates using a device ejecting a spray comprising sirolimus and/or analogs of sirolimus
under gas pressure that aerosolizes upon exiting a tubular extension rod housing. Fujita et al.,
Method For Remote Delivery Of An Aerosolized Liquid. United States Patent No. 5,722,950
(herein incorporated by reference). Alternatively, a spray may be generated by slits through
the walls of an implanted medical-surgical tube such as a tracheal tube, thoracic or trocar
catheter. Preferably, the spray may be an aerosol, coarse spray or liquid stream as determined
by the number and size of piercings through the tube wall into the lumen of the tube.

Sheridan D., Medico-Surgical Tube Including Improved Means For Administering Liquid Or
Gas Treatment. United States Patent No. 5,207,655 (herein incorporated by reference).
In one embodiment, the present invention contemplates a spray tip 71, wherein a medium is
nebulized by a small orifice 72 (see Figure 6). Preferably, said spray 71 comprises a luer
lock 73 thus allowing compatibility with any standard medical connectors.
An exemplary endoscope shaft 44 (Figure 7) may be used during laparoscopic or
arthroscopic procedures comprising a viewing optical fiber 48 and a first lumen 47 and a
second lumen 46. First lumen 47 could be used for operating a surgical cutting tool (not
shown) and second lumen 46 can be used for administering sirolimus and analogs of sirolimus
12 into the surgical site using an endoscopic delivery catheter 43.
In another embodiment, a catheter comprises a common lumen for both a surgical
cutting device and for the delivery of a medium comprising sirolimus, tacrolimus and analogs
of sirolimus. In one embodiment, sirolimus, tacrolimus or an analog of sirolimus may be
administered in the form of a liquid spray, pourable liquids, squeezable liquids, a foam, a gel,
a hydrogel, or sheet of material. In another embodiment, the sirolimus compounds are in the
form of microparticles as described herein. In one embodiment, a medium comprising said
microparticles decreases post-surgical complications by reducing scar tissue formation
following either or both a laparoscopy or arthroscopy procedure.
In another embodiment, an endoscopic delivery catheter is inserted through an organ
lumen 46 to deliver a medium to a closed surgical site. Figure 8 shows one embodiment of a
typical endoscopic delivery catheter. A female Luer lock adapter 81 is connected to a
reservior (not show) that allows a medium comprising sirolimus, tacrolimus or an analog of
sirolimus to flow through the catheter lumen 82 and exit the catheter at side ports 83.
The present invention contemplates a method comprising pouring a medium into an
open surgical site. In one embodiment, the liquid medium is poured from a hand-held
container wherein a flexible tube is capable of directing the flow of the liquid medium.
Preferably, said hand-held container includes, but is not limited to, a bottle, a dish, or a
mixing tray. In another embodiment, the liquid medium is poured from a fixed container that
may be tilted by remote control or manually a medical assistant. Preferably, said fixed
container includes, but is not limited to, an applicator tube with a valve for controlling the

flow of the medium. In another embodiment, the medium is applied from a tube into an open
surgical site. In another embodiment, the medium is applied by squeezing a squeeze bottle.
Bioadhesives
One aspect of the present invention contemplates a bioadhesive medium comprising
sirolimus and analogs of sirolimus. Preferably, various embodiments of a bioadhesive
medium comprise a biocompatible and biodegradable patch designed for use inside a living
organism. In one embodiment, a bioadhesive patch releases a constant compound dose over a
period of at least 1 day to 6 months. One of skill in the art would recognize that this
embodiment is superior to most conventional transdermal patches currently available for the
epidermal layer of the skin. Although it is not necessary to understand the mechanism of an
invention it is believed that a bioadhesive patch will heal a wound faster than applying a
topical medication that acts locally for only a short time. Additionally, long duration
bioadhesive patches do not have the inconvenience and cost of adding more medication for
multiple dressing changes. Some bioadhesives are applied using the techniques of liquid
administration as defined above.
One embodiment of the present invention contemplates a bioadhesive comprising
sirolimus and analogs of sirolimus in combination with a wound healing agent comprising a
dental enamel matrix. Gestrelius et al., Matrix Protein Compositions For Wound Healing.
United States Pat. No. 6,503,539 (herein incorporated by reference). Alternatively,
Liquiderm™ adhesive and Dermabond® Topical Skin Adhesive (Closure Medical Corporation)
are also compatible with the present invention. Dermabond® adhesive is known as a viable
alternative to sutures and staples in closing incisions and lacerations. Liquiderm™ adhesive is
brushed on the wound, seals the wound from dirt and germs thereby creating a healing
environment.
One embodiment of the present invention contemplates a bioadhesive comprising
sirolimus and analogs of sirolimus and an adhesive material consisting of a mixture of
hemoglobin and albumin in a solution of glutaraldehyde. Preferably, the coating functions as
both a repository for controlled compound release and provides external vascular structural
support following surgery. Ollerenshaw et al, Vascular Coating Composition, United States
Patent No. 6,372,229 (herein incorporated by reference).

One aspect of the present invention contemplates a method of anastomoses using a
bioadhesive comprising sirolimus and analogs of sirolimus. Bioadhesives are known to be
useful for anastomoses. Black et ah, Sutureless Anastomotic Technique Using A Bioadhesive
And Device Therefore, United States Patent No. 6,245,083 (herein incorporated by reference)
The impregnation of bioadhesives with sirolimus and analogs of sirolimus, however, to reduce
post-surgical scarring is novel.
In one embodiment, the present invention contemplates a method of joining organs, at
least one of which has an internal cavity, using a bioadhesive comprising cross-linked
proteinaceous materials and a compound selected from the group consisting of sirolimus,
tacrolimus and analogs of sirolimus. Preferably, the organs are held in apposition {i.e., by
hand or a surgical device) and the organs are joined together using a compound impregnated
bioadhesive of the present invention. This joining is facilitated by the creation of apertures by
cutting the wall of the organ to allow the introduction of one organ into the other. When the
apertures are held together, an anastomosis site is formed at the interface of the two organs to
which the bioadhesive of the present invention is applied. For example, a device can be
attached to each organ through the use of expandable balloons that become stabilized within
the organs when they are inflated. The expandable balloons can be attached to one another by
a means extending through the apertures. Hence, an arteriotomy site is dilated while holding
the oragns to be anastomosed in contact while the bioadhesive is applied. The amount of
bioadhesive used is sufficient to seal the joined organs so that the apertures communicate,
thereby enabling liquids and compounds to move from one organ to the other through the
apertures. Once the bioadhesive sets, the cavities of the two organs can communicate through
the joined apertures.
The present invention contemplates a bioadhesive suitable for use in an anastomoses
that is non-toxic, has the capability to adhere to biological tissues, reaches stability quickly
(typically within about 30 seconds to about 5 minutes), preferably set in wet conditions, bonds
to both biological tissues and synthetic materials, and provides sufficient strength to stabilize
organs having undergone an anastomosis joining. Preferably, bioadhesive compositions
comprising sirolimus and analogs of sirolimus wherein said composition consists of a
proteinaceous material and a cross-linking agent are contemplated by this invention for

anastomoses. Kowanko N., Adhesive Composition And Method, United States Pat. No.
5,385,606 (hereby incorporated herein by reference). The '506 bioadhesive compositions
contains two components: i) from 27-53% by weight proteinaceous material; and ii) di- or
polyaldehydes in a weight ratio of one part by weight to every 20-60 parts of protein present.
To produce the bioadhesive, the two parts are mixed and allowed to react on the surface to be
bonded. Bond formation is rapid, generally requiring less than one minute to complete. The
resulting adhesion is strong, capable of providing bonds with tear strengths of between
400-1300 g/cm2.
Another suitable bioadhesive compatible with the present invention are made by the
condensation of a carboxylic diacid with a sulphur-containing amino acid or one of its
derivatives. These products contain reactive thiol SH functions which may oxidize to form
disulfide bridges, leading to polymers which may or may not be crosslinked. Constancis et
al, Adhesive Compositions For Surgical Use. United States Patent No. 5,496,872 (herein
incorporated by reference).
One embodiment of the present invention contemplates the extrusion of a double
component bioadhesive comprising sirolimus and analogs of sirolimus. In one embodiment,
the invention relates to a method of joining, or anastomosing, tubular organs in a side-to-side
or end-to-side fashion using bioadhesive. For example, a double component bioadhesive of
the '506 patent may be applied through an extruding device having a mixing tip. In one
embodiment, a bioadhesive is extruded onto the interface of the two organs in an open
surgical field where medical personnel have free and open access to the anastomosis site. In
another embodiment, a bioadhesive may be applied by a catheter directed through an
endoscope {infra).
The details of the anastomosis embodiment can be exemplified in terms of performing
coronary bypass surgery. One embodiment of the present invention contemplates a method
for anastomosis of the internal mammary artery (hereinafter "IMA"), also called the internal
thoracic artery, to a branch of the left coronary artery comprising; i) isolating an IMA from
the chest wall; ii) clamping at a location proximal to the intended site of anastomosis; iii)
incising said IMA distal to the intended site of anastomosis; iv) elevating a host artery:
v) incising said IMA thus creating a first aperture; vi) isolating said host artery; vii) incising

said host artery thus creating a second aperture; viii) inserting a double balloon catheter in
said IMA such that said catheter passes through said first aperture and protrudes into said
second aperture; ix) inflating a first balloon of said catheter within said host artery such that
said second aperture is stabilized; x) positioning said first and second apertures such that they
are directly apposed; xi) inflating a second balloon of said catheter within said IMA such that
said first aperture is stabilized; xii) applying a bioadhesive comprising sirolimus and analogs
of sirolimus around said apposed first and second apertures such that a sufficient strength is
reached to maintain an anastomosis; xiii) removing said catheter from said anastomosis; and
xiii) ligating said anastomosis.
One embodiment of the present invention contemplates a bioadhesive patch comprising
a hydrogel (infra) and a compound selected from the group consisting of sirolimus, tacrolimus
and analogs of sirolimus. In a clinical setting, medical personnel would apply the patch
containing the compound to a wound, covering it with a bandage. The bandage maintains
contact of the hydrogel with the wound and prevents the hydrogel from drying out.
Alternatively, a cytostatic and antiproliferative compound (i.e., for example, sirolimus,
tacrolimus and analogs of sirolimus) may be incorporated directly into the hydrogel or
attached to microparticles, wherein said microparticles are residing within the hydrogel. In
one embodiment, a bioadhesive comprising microparticles provide a controlled release
medium of said compound.
Bioadhesives are known to comprise fibrin glues, cyanoacrylates, calcium
polycarbophil, polyacrylic acid, gelatin, carboxymethyl cellulose, natural gums such as karaya
and tragacanth, algin, chitosan, hydroxypropylmethyl cellulose, starches, pectins or mixtures
thereof. Alternatively, the adhesives may be combined with a hydrocarbon gel base,
composed of polyethylene and mineral oil, with a preselected pH level to maintain gel
stability.
In one embodiment an adhesive gel is adjusted to a preselected pH wherein the gel
comprises microcapsules. Adhesive biogel system is then placed into a surgical site under
conditions such that the active ingredient is delivered.
Foams
One aspect of the present invention contemplates a medium comprising a foam and

sirolimus and analogs of sirolimus. It is well known in the art that a foam medium is
generally produced from a previously manufactured hydrogel or gel. Therefore, one of skill
in the art will understand that any hydrogel medium disclosed herein may be converted into a
counterpart foam medium. Many different compositions of foams are known in the art,
therefore, the following is only intended as one example of a foam contemplated by the
present invention. It is not intended that the present invention be limited by this type of
foam.
One embodiment of the present invention contemplates a foam comprising a
water-swellable polymer gel and sirolimus and analogs of sirolimus produced by a general
process of lyophilizing a gel swollen with water, or by introducing bubbles into the internal of
the gel. Preferably, a method for preparing a foam comprising introducing bubbles into the
internal of the gel includes processes disclosed in British Patent No. 574,382, Japanese Patent
Laid-Open Nos. Hei 5-254029, 8-208868 and 8-337674 and Japanese Unexamined Patent
Publication No. Hei 6-510330, and the like. Particularly, when a foam of the water-swellable
gel of the present invention is prepared by the process below, there is obtained a foam of a
water-swellable polymer gel having higher water absorbability and higher stability as
compared to those foams disclosed in those publications.
One example of a method for preparing a foam comprising: i) introducing bubbles into
the internal of a gel comprising a compound selected from the group consisting of sirolimus,
tacrolimus and analogs of sirolimus, ii) introducing bubbles into an esterified polysaccharide
solution or a polyamine solution such that foaming occurs, and iii) contacting said foamed
solution with said polyamine solution or said esterified polysaccharide, respectively, to cause
gelation. In another example, a method comprises; i) introducing bubbles into a mixed
solution of an esterified polysaccharide and a polyamine such that foaming occurs, and ii)
completing gelation.
In another embodiment, a method for preparing a foam comprises, i) introducing
bubbles into a solution comprising a compound selected from the group consisting of
sirolimus, tacrolimus and analogs of sirolimus that is capable of foaming; ii) adding a foaming
agent such that a water-insoluble gas is generated and foaming occurs. Preferably, said gas
generation results from heating or a chemical reaction using, for instance, but not limited to,

ammonium carbonate, azodicarbonamide, p-toluenesulfonyl hydrazide, butane, hexane, and
ether. Any method to prepare a foam may further comprise mechanically stirring the solution,
thereby diffusing a fed gas into the aqueous solution to foam; and the like.
Any method to prepare a foam may further comprise an ionic or non-ionic surfactant
(i.e., a "surface active agent"), which is a bubble-forming agent, as occasion demands, in
order to stabilize the foam. In one embodiment, an ionic surfactant includes, for instance,
anionic surfactants such as sodium stearate, sodium dodecyl sulfate, a-olefinsulfonate and
sulfoalkylamides; cationic surfactants such as alkyldimethylbenzylammonium salts,
alkyltrimethylammonium salts and alkylpyridinium salts; and amphoteric surfactants such as
imidazoline surfactants: In another embodiment, a non-ionic surfactant includes, for instance,
polyethylene oxide alkyl ethers, polyethylene oxide alkylphenyl ethers, glycerol fatty acid
esters, sorbitan fatty acid esters, sucrose fatty acid esters, and the like.
Low molecular weight surfactants are known for irritating and denaturing living tissue
or a physiologically active substance (i.e., an enzyme or the like). Preferably, non-toxic
surfactants are contemplated for foam embodiments of the present invention. Foams
contemplated by the present invention comprise a non-toxic surfactant, that are a collection of
complex molecules aggregating at the bubble's surfaces. Preferably, such surfactants include,
but are not limited to, fats or proteins in edible foams or chemical additives in shaving cream.
Although it is not necessary to understand the invention, it is believed that surfactants act by
preventing surface tension from collapsing the foam structure by keeping the bubbles separate
and repelling water from their surfaces. Foams sprayed from hand-held canisters are capable
of expanding to about 100 times their liquid volume as air is drawn into the spray. An
advantage of a foam over a liquid is that the foam fills crevices and other elusive hiding
places as the expansion process occurs.
Although it is not necessary to understand an invention, it is believed that an esterified
polysaccharide itself exhibits amphipathic properties and functions as a bubble-forming agent
for stabilizing the gas-liquid interface. Consequently, in some embodiments a surfactant may
not be necessary in the presence of esterified polysaccharides. Since an esterified
polysaccharide has reactivity in addition to the amphipathic property, the esterified
polysaccharide can be referred to as a "reactive surfactant polysaccharide."

In some embodiments, surfactants may also be, but not limited to, a protein such as
albumin, gelatin or albumin, or lecithin.
In one embodiment, a method for preparing a foam further comprises adding a bubble
stabilizer. Preferably, bubble stabilizers include, but are not limited to, a higher alcohol such
as dodecyl alcohol, tetradecanol or hexadecanol; an amino alcohol such as ethanolamine; a
water-soluble polymer such as carboxymethyl cellulose; and the like. Alternatively, bubble
stabilizers may be polysaccharides comprising natural polysaccharides such as agarose,
agaropectin, amylose, amylopectin, arabinan, isolichenan, curdlan, agar, carrageenan, gellan
gum, nigeran and laminaran. While it is not necessary to understand an invention, it is
believed that bubble stabilizers prevent the disappearance of bubbles prior to the completion
of crosslinking.
In one embodiment, the present invention contemplates a pressurized canister
comprising a foam and a compound, such as, but not limited to, sirolimus, tacrolimus and
analogs of sirolimus. As depicted in Figure 9 a pressurized foam canister 80 has a generally
cylindrical body. Foam canister 80 includes a movable dispensing valve 75 coupled thereto
that is accessed by finger aperature 62. Valve 75 is constructed in accordance with
conventional fabrication techniques and defines an upwardly extending valve passage 76 and a
laterally extending ledge 77. Valve 75 is operable to discharge the pressurized foam contents
through valve passage 76. A generally conical cap 60 defines a nozzel aperture 61 at its apex
and a downwardly extending nozzle passage 65. Valve 75 also extends partially into nozzle
passage 65 within cap 60.
Gels
A hydrdgel medium comprises a three-dimensional networks of hydrophilic polymers,
either covalently or ironically cross-linked, which interact with aqueous solutions by swelling
and reaching an equilibrium. Compounds, such as, but not limited to, sirolimus and analogs
of sirolimus, can be added to a hydrogel medium during the manufacturing process. Hydrogel
medium technology encompasses many different types of compositions, therefore, the term
"hydrogel" does not refer to any specific composition but identifies a composition having
specific properties. For example, hydrogels may provide controlled release of drug
compounds included in them by providing physical barriers or through chemical attachment of

the drug to the hydrogel.
Hydrogels are primarily characterized by having an ability to swell in aqueous
solutions. Swelling ratios and solubility are controlled by the specific composition of the
hydrogel. Higher swelling ratios result in a greater release rate of an incorporated compound
that is attached to or contained within the hydrogel. Although it is not necessary to
understand the mechanism of an invention, it is believed that a high swelling ratio results in
more open structure within the hydrogel and more closely mimics living tissue, therefore
facilitating the process of diffusion between the hydrogel and the tissue. High swelling ratios
are also related to the overall hydrophilicity of the hydrogel composition, and provide for
better absorptive properties.
In one embodiment, the present invention contemplates a hydrogel matrix having the
capability to provide controlled release of a compound prepared by: i) adding heparin (400
mg, 0.036 mmole) to 750 mis of double distilled water at 4°C; ii) adding human serum
albumin (550 mg, 0.0085 mmole) 1.0 ml double distilled water at 4°C, and hi) adding
N-(3-dimethylaminopropyl)- N-ethylcarbodiimide (i.e., EDC solution; 94 mg) to 250 ml
double distilled water at 4°C. The heparin solution, along with the 1 ml of the albumin
solution are first mixed within a 2 ml polyethylene-polypropylene syringe containing a small
stir bar and a desired concentration of a compound (i.e., for example, sirolimus).
Subsequently, the EDC solution is added to form the final mixture. All steps are carried out
at 4°C.
After 24 hours, a hydrogel is removed from the syringe by swelling as the syringe is
placed in toluene. After the albumin-heparin hydrogel is extruded from the syringe, the
hydrogel is then equilibrated with phosphate buffered saline to remove uncoupled material.
The release rate of the attached compound may be controlled by varying the amount of
heparin present in the matrix.
One embodiment of the present invention contemplates a hydrogel laminate comprising
sirolimus and analogs of sirolimus and crosslinked hydrophilic-adhesive polymers. Such
compositions form absorbent products such as bandages. Preferably, hydrogel polymers are
generally synthetic polyvinylpyrrolidone, polyethyleneoxide, acrylate, and methacrylate and
copolymers thereof. Kundel, Hydrogel Laminate, Bandages and Composites And Methods

For Forming The Same, United States Pat. No. 6,468,383 (herein incorporated by reference).
Alternatively, hydrogels compatible with the present invention may be formed by crosslinking
carbohydrates, such as dextran, with maleic acid or hyaluronic acid with polyvinyl chloride.
Kim et al, Dextran-Maleic Acid Monoesters And Hydrogels Based Thereon. United States
Patent No. 6,476,204; Giusti et al, Biomaterial Comprising Hyaluronic Acid and Derivatives
Thereof In Interpenetrating Polymer Networks (IPN). United States Patent No. 5,644,049
(both herein incorporated by reference).
Another embodiment contemplates a hydrogel medium comprising hyaluronic acid
capable of controlled release of sirolimus and analogs of sirolimus. While these compositions
are disclosed as topical and injectable polymer solutions, the present invention contemplates a
hyaluronic acid polymer solution within a hydrogel to time-release the delivery of compounds,
such as, but not limited to, sirolimus and analogs of sirolimus within the body. Drizen et al.,
Sustained Release System. United States Patent No. 6,063,405 (herein incorporated by
reference).
One aspect contemplated by the present invention comprises a hydrogel medium
comprising sirolimus or analogs of sirolimus, wherein said hydrogel has a controlled gelation
time. In one embodiment, the hydrogel is made of one or more synthetic and/or natural
water-soluble polymers, and one or more divalent or multivalent cation containing or releasing
compounds. At least one of the polymer monomers is an acid or a salt thereof that is capable
of reacting with the divalent or multivalent cation to form intermolecular polymer ionic
crosslinks. Such hydrogels are discussed in detail relating to use for tissue culture
. scaffolding. Ma P.X., Ironically Crosslinked Hydrogels With Adjustable Gelation Time.
United States Patent No. 6,497,902 (herein incorporated by reference). Specifically,
controlled gelation time is taught as a function of: i) cation solubility; ii) cation concentration;
iii) mixture/ratio of cation containing compounds; iv) polymer concentration; and v) gelation
temperature.
In one embodiment, the present invention contemplates the administration of a
hydrogel comprising sirolimus and analogs of sirolimus to an open surgical site. In another
embodiment, the present invention contemplates the administration of a hydrogel comprising

sirolimus and analogs of sirolimus to a closed surgical site via a catheter (i.e., during
laparoscopic procedures) that transitions into a gel upon contact with living tissue. In one
embodiment, a micelled hydrogel core serves as a reservoir of sirolimus and analogs of
sirolimus. In another embodiment, a hydrogel comprises microparticles attached to sirolimus
and analogs of sirolimus. Sirolimus and analogs of sirolimus are contemplated by the present
invention as pharmacologically effective in reducing scar tissue and improving the healing of
wounds or surgical incisions.
One embodiment of the present invention contemplates a controlled release hydrogel
medium comprising sirolimus and analogs of sirolimus formed by crosslinking a protein (i.e.,
albumin, casein, fibrinogen, y-globulin, hemoglobin, ferritin and elastin) with a polysaccharide
(i.e., heparin, heparan, chondroitin sulfate and dextran). Determinative factors regulating
compound release from a hydrogel medium is: i) gel composition; ii) crosslinking degree; and
iii) gel surface treatments. Specifically, it is known that hydrogel releasable compounds
include hormones, cytostatic agents, antibiotics, peptides, proteins, enzymes and
anticoagulants. Feijen J., Biodegradable Hydrogel Matrices For the Controlled Release Of
Pharmacologically Active Agents. United States Patent No. 4,925,677 (herein incorporated by
reference). Alternatively, controlled release of compounds from a hydrogel medium
contemplated by the present invention is possible by inserting hydrolyzable spacers between
polymer crosslinks. In one embodiment, a hydrogel degradation rate is contemplated to be
modified to provide dissolution rates from 1 day to 6 months. Hennink et al., Hydrolyzable
Hydrogels For Controlled Release. United States Patent No. 6,497,903 (herein incorporated
by reference).
Alternatively, a hydrogel medium may act as a compound repository in their own right
wherein diffusion creates a time-release delivery of a compound into the surrounding tissue.
Kennedy et al., Semisolid Therapeutic Delivery System And Combination Semisolid,
Multiparticulate, Therapeutic Delivery System. United States Patent No. 6,488,952 (herein
incorporated by reference). In one embodiment, the present invention contemplates a
hydrogel comprising a liposome comprising sirolimus and analogs of sirolimus covalently
attached to a medical device, such as, for example, a wound dressing. Preferably, a hydrogel

medium contemplated by this invention comprises a material selected from the group
consisting of gelatins, pectins, collagens and hemoglobins. DiCosmo et al., Compound
Delivery Via Therapeutic Hydrogels, United States Patent No. 6,475,516 (herein incorporated
by reference). In one particular embodiment, a hydrogel comprises microparticles containing
sirolimus or an analog of sirolimus.
One embodiment of the present invention contemplates a method providing a medical
device comprising a catheter capable of placing a hydrogel comprising sirolimus and analogs
of sirolimus at a closed surgical site. Sahatjian et al., Compound Delivery, United States
Patent No. 5,674,192 (herein incorporated by reference). Preferably, said hydrogel comprises
a second compound designed as a wound healing agent such as, but not limited to, dental
enamel matrix. Gestrelius et al., Matrix Protein Compositions For Wound Healing. United
States Pat. No. 6,503,539 (herein incorporated by reference).
One aspect of the present invention contemplates thermo-reversible gel technology
based on the use of biocompatible poloxamers made up of polyoxyethylene and
polyoxypropylene units. Preferably, these poloxamers comprise any polymer or copolymer
sold under the trademarks Pluronics® or Tetronics®. A Tetronic ® gel-forming macromer
contains four covalently linked polymeric blocks, wherein at least one polymeric block is
hydrophilic, linked by a common crosslinkable group and is disclosed as a thermal gelling
drug delivery device. United States Patent No. 6,410,645 To Pathak et al. (herein
incorporated by reference). These gels are discussed as having thermosensitivity and
lipophilicity, and may be used to administer drugs and tissue coatings for medical
applications. Other Tetronic ® polyols, having hydrophobic polymeric blocks, are known as
drug delivery devices. United States Patents 4,474,751; 4,474,752; 4,474,753; and 4,478,822
To Haslam et al. (all herein incorporated by reference).
In one embodiment, poloxamer 407 {i.e., Pluronics® F-127) is a primary ingredient
and can be manufactured in a variety of formulations with specific physical and chemical
properties. The most significant physical characteristic of thermo-reversible gels is an ability
to change from- a liquid to a gel upon wanning to body temperature. This characteristic
allows for manipulation of the polymer product in its liquid state and conversion to a desired

solid state (i.e., a gel) in or on the body of the patient. One specific advantage of
administering a thermal gel in a liquid state includes molding to body/tissue contours before
gelling in place. Thus, the thermal gel maintains contact with the tissue surface and serves as
a physical, protective barrier in addition to serving as a carrier for drug delivery to adjacent
tissues. Typically, thermal gels are comprised of materials known to be non-toxic,
non-irritating and pharmacologically inert. Furthermore, thermal gels dissolve in the body and
are cleared by the normal excretory processes.
The present invention contemplates a biocompatible thermal gel medium comprising
sirolimus and analogs of sirolimus attached to microparticles. In one embodiment, the
microparticles are capable of controlled release of the sirolimus and analogs of sirolimus. In
one embodiment, the thermal gel medium comprises a polymer gel, such as, but not limited to
Flogel® (Alliance Pharmaceutical Corp). Preferably, polymer gels such as FloGel® are applied
to tissues and organs as a chilled liquid that solidifies into a gel as it warms to body
temperature, creating a physical barrier that holds the microspheres in place while the thermal
gel and the microspheres bioerode and the cytostatic compound is released such that excess
scar tissue is prevented.
Xerogels
One aspect of the present invention contemplates a device and method for long-term
controlled release of a medium comprising sirolimus, tacrolimus and analogs of sirolimus. In
one embodiment, the medium comprises a xerogel, exemplified by the commercially available
product Xerocell™ (Gentis, Berwyn, PA). Xerogels comprise a plurality of microscopic air
bubbles suffused in a glassy matrix. In one emboidment, the present invention contemplates a
controlled release medium comprising a xerogel and sirolimus, tacrolimus and/or analogs of
sirolimus. Preferably, the xerogel allows complete control over a controlled release profile
from approximately a few hours to more than a year.
One aspect of the present invention contemplates a method comprising placing a
xerogel comprising sirolimus, tacrolimus and analogs of sirolimus at or near a surgical site.
In one embodiment, said surgical site heals over and around the xerogel. In one embodiment,
the xerogel provides a controlled release the sirolimus, tacrolimus and/or analogs of sirolimus
such that surgical scar and/or adhesion tissue formation is reduced.

. One aspect of the present invention contemplates surgical dressings and surgical tapes
comprising a medium and sirolimus and analogs of sirolimus. Illustrative examples of such
dressings and tapes include, but are not limited to, sheets of material, surgical swabs, gauze
pads, closure strips, compress bandages, surgical tape, etc. For example, one embodiment of
the present invention contemplates a laminated composite comprising a first nonwoven fiber
layer, an elastic layer, a melt blown adhesive fiber layer, and a second nonwoven fiber layer,
wherein said composite comprises sirolimus and analogs of sirolimus impregnated into said
second nonwoven layer. Menzies et al, Laminated Composites, United States Patent No.
6,503,855 (herein incorporated by reference).
In one embodiment (Figure 10), the present invention contemplates a biocompatible
sheet of material or mesh comprising sirolimus and analogs of sirolimus impregnated (i.e., ,
attached) into, coated onto or placed onto a material sheet or mesh. Such sheets of material
may placed between internal body tissues to prevent the formation of post-operative adhesions
and/or scar tissue. In one embodiment, the sheet of material is biodegradable (Surgicel™,
Johnson & Johnson). In another embodiment, said sheet of material comprises a surgical
suture. In another embodiment said sheet of material comprises a surgical staple. In another
embodiment, said sheet of material comprises an eye buckle. In another embodiment, said
sheet of material comprises a cylindrical tube.
In one embodiment, the present invention contemplates a moist dressing product
comprising a medium of sirolimus and analogs of sirolimus. Preferably, these dressings
consist of a flexible film having a polyurethane gel core. Although it is not necessary to
understand the mechanism of an invention, it is believed that moist dressing products reduce
the formation of a hard scab and reduces the likelihood of scarring. For example, these
dressings may include, but are not limited to, those currently marketed as Elastoplast® (Active
Gel Strips; Beiersdor, Inc.).
In one embodiment, the present invention contemplates a semipermeable membrane
formed from a unique blend of silicone and polytetrafluoroethylene (PTFE) and a medium of
sirolimus and analogs of sirolimus. Although it is not necessary to understand the mechanism
of an invention, it is believed that the PTFE provides an internal reinforcing mechanism,

thereby creating very thin sheets of soft silicone with significantly enhanced physical strength.
For example, these dressings may include, but are not limited to, those currently marketed as
Silon-IPN™ (Bio Med Sciences).
In one embodiment, the present invention contemplates dressings comprising a
polyurethane membrane-matrix on a semi-permeable thin-film backing and sirolimus and
analogs of sirolimus. Preferably, the hydrophilic membrane contains a cleanser, a moisturizer
and a super-absorbent starch co-polymer. Although it is not necessary to understand the
mechanism of an invention, it is believed that eliminates the need for manual debridement and
cleaning during dressing changes is eliminated and reduces patient discomfort and the time
and cost of dressing changes. For example, these dressings may include, but are not limited
to, those currently marketed as PolyMem® (Ferris Mfg., Inc.).
In one embodiment, the present invention contemplates closure strips comprising
sirolimus and analogs of sirolimus. Preferably, said skin closures are useful in a method to
provide skin closure following intra-abdominal operations. Alternatively, these closures may
be used with any traditional sutures of sutures coated with sirolimus and analogs of sirolimus.
Although it is not necessary to understand the mechanism of an invention, it is believed that
the advantages of skin closures contemplated by the present invention are: i) lower rates of
infection and over-all morbidity; ii) a lower cost; iii) a reduction in time in the operating
room when compared with conventional methods; and iv) avoidance of foreign body
granulomas, strangulation, tissue necrosis and cellulitis. Pepicello et al. Five Year Experience
With Tape Closure Of Abdominal Wounds. Surg Gynecol Obstet 169:310-4 (1989).
Marker Agents
The present invention contemplates incorporating any color as a marker agent into any
medium discussed herein. In one embodiment, a desired colored medium comprises a marker
comprising a colored dye or stain such as the blue dye "Brilliant Blue R", also known as
"Coomassie™ Brilliant Blue R-250" (distributed as "Serva Blue"; Serva) The resulting
medium has a blue color that provides a good contrast to the color of body tissues, making
the medium easy to see during surgery. In another embodiment, the present invention
contemplates a gel, film or spray made up of two liquids which comprise sirolimus and
analogs of sirolimus, that when sprayed together, solidify to form a bright colored material

which breaks down gradually over about a week.
One embodiment of the present invention contemplates a method providing a
biocompatible and biodegradable microsphere or hydrogel having a coloring marker agent
such that medical personnel are capable of adequately covering an intended region where scar
tissue and/or adhesions might form.
The present invention also contemplates incorporating a radio-opaque marker into any
medium discussed herein. In one embodiment, said radio-opaque marker comprises a barium
compound. In one embodiment, said radio-opaque marker is visualized using X-ray
fluoroscopy.
For any of the applications described herein, the systemic application of one or more
of the cytostatic anti-proliferative agents that have been described could be used conjunctively
to further minimize the creation of scar tissue. The systemic application could be by mouth,
by injection, or by any other well known means for placing a compound systemically into a
human body.
Although only the use of certain compounds, such as, sirolimus and analogs of
sirolimus, and those capable of binding to the mTOR protein and/or interrupting the cell cycle
in the GO or Gl phase has been discussed herein, it should be understood that supplemental
pharmaceutical compounds may be provided to improve the outcome for the patients.
Specifically, an antibiotic, and/or analgesic, and/or anti-inflammatory agent could be added to
prevent infection and/or to decrease pain. It is further understood that any patient in whom
sirolimus and analogs of sirolimus is used in combination with at least one supplemental
pharmaceutical compound may have an improved response if sirolimus and analogs of
sirolimus is also given as a conventional administration.
Various other modifications, adaptations, and alternative designs are of course possible
in light of the above teachings. Therefore, it should be understood at this time that within the
scope of the appended claims, the invention can be practiced otherwise than as specifically
described herein.

Experimental
The following examples serve to illustrate certain preferred embodiments and aspects
of the present invention and are not to be construed as limiting the scope thereof.
In the experimental disclosure which follows, the following abbreviations apply: g
(gram); mg (milligrams); ug (microgram); M (molar); mM (milliMolar); fiM (microMolar);
ran (nanometers); L (liter); ml (milliliter); pi (microtiters); CC (degrees Centigrade); m
(meter); sec. (second).
Example I
A Controlled Release Microsphere For Hvdrophobic Compounds
This example describes the production of a microsphere capable of administering
sirolimus in controlled release manner.
A controlled release microsphere pharmaceutical composition is made that is burst-free
and provides a sustained programmable release of a sirolimus compound over a duration of 24
hours to 100 days made in accordance with United States Patent No. 6447796 To Vook et al.
(herein incorporated by reference). These microspheres are particularly suited for
hydrophobic drugs by using a blend of end-capped and uncapped biocompatible,
biodegradable poly(lactide-co-glycolide) copolymers (PLGA). The end-capped polymers have
terminal residues functionalized as esters and the uncapped polymers have terminal residues
existing as carboxylic acids.
PLGA copolymers contemplated by this Example has a molecular weight ranging from
10 to 100 kDa are in a 50:50 ratio, although one skilled in the art would understand that other
ratio's are also possible. Briefly, well known solvent evaporation techniques are used to
prepare sirolimus/PLGA microspheres in a range of 0.1 - 2.0 mg of sirolimus per 100 mg
PLGA. The evaporation technique is expected to result in microsphere core loads of 10%,
20%, 40%, and 50% of a theoretical maximum. Empirical testing is performed to determine
the proper ratios of sirolimus and PLGA copolymer concentrations that result in these
predicted core loading efficiencies.
For example, a useful protocol is as follows:
1) Pre-heat water bath to 15°C

2) Prepare 1% poly-vinyl alcohol solution in distilled water.
3) Prepare a 1% poly-vinyl alcohol solution in methylene chloride-saturated
distilled water.

3) Co-dissolve appropriate amounts of sirolimus and PLGA in 3.5 g methylene
chloride.
4) Add the PLGA-sirolimus solution to 25 ml of the 1% poly-vinyl alcohol
solution in methylene choloride-saturated distilled water.
5) Homogenize the mixture at 10,000 rpm for 30 seconds in a 50 ml centrifuge
tube.
6) Add the homogenized mixture to 500 ml of the 1% poly-vinyl alcohol solution
in distilled water.
7) Stir at 650 rpm for 1/2 hour at 15°C.
8) Stir at 650 rpm for 4 hours at 25°C.
9) Collect microspheres by filtration.
10) Wash collected microspheres.
10) Vacuum dry collected microspheres overnight.
Microspheres are expected to show an average diameter range of between 2.5-200 jim,
prefereably between 4.0-75 \im, and more preferably between 5.0-10.0 \im. Release rates in
relationship to core loading capacity are expected as: i) 40.19% sirolimus release in 10 days
using a 10% core load; ii) 71.58% sirolimus release in 6 days using a 20% core load; iii)
48.09% sirolimus release in 6 days using a 40% core load; and iv) 39.84% sirolimus release
in 6 days using a 50% core load. Administration of sirolimus containing microspheres
prepared according to this method may be performed by any method contemplated herein.
Example II
Liposome Encapsulation
This example describes a method to prepare liposomes that encapsulate sirolimus.
Multilammelar vesicles (i.e., liposomes) are prepared from egg phosphatidylcholine
(EPC) and cholesterol (Ch) (ratio 4:3). Specifically, a preliposomal lipid film will be

obtained by drying under nitrogen atmosphere a mixture of EPC (14.4 mg = 18.3 mnoles),
5.6 mg cholesterol (13.7 jimoles) and 0.1 mg sirolimus in a nonpolar organic solvent such as
dichloromethane or chloroform. The resulting dry lipid film is then converted into a
liposomal suspension of the multilammelar vesicles encapsulating the sirolimus by hydrating
the dry lipid film with 1 ml isotonic phosphate buffer pH 8.1, and smooth shaking of the
suspension during formulation. Finally, sorbitol is then added to the suspension in an
amount of 1% wt/volume, at a molar ratio of sorbitol to phospholipid of 3:1.
The final liposomal suspension is then freeze dried at -25 °C by direct immersion in
denatured ethanol. The association (i.e., encapsulation efficiency) of sirolimus with 1 ml of
the liposomal suspension with 1 ml of the liposomal suspension before and after freeze drying
is expected to be approximately 80%.
Example III
A Hvdrogel Composition
This example provides a composition where sirolimus is incorporated into a hydrogel
such that the sirolimus is released by diffusion.
This hydrogel composition will incorporate and retain significant amounts of H2O, and
eventually reach an equilibrium content in the presence of an aqueous environment. Glyceryl
monooleate (i.e., GMO) is described herein, however on skilled in the art will recognize that
many polymers, hydrocarbon compositions and fatty acid derivatives having similar
physical/chemical properties with respect to viscosity/rigidity are capable of producing
hydrogels for purposes of this invention.
First, the GMO is heated above its melting point (i.e., 40°C - 50°C). Second, a warm
aqueous-based buffer (i.e., an electrolyte solution) such as phosphate buffer or normal saline
or a semi-polar solvent containing the desired concentration of any sirolimus suspension or
water soluble sirolimus derivative as discussed herein, is added to produce a three-dimensional
hydrogel composition.
The selection of GMO as a gel polymer is advantageous due to its amphipathic
properties. Specifically, GMO will provide a predominantly lipid-based hydrogel, thereby
incorporating lipophilic compounds such as sirolimus.

At room temperature (i.e., 20°C- 25°C) this hydrogel will exist in a lamellar phase
consisting of approximately 5%-15% H2O and 95% - 85% GMO. This lamellar phase is a
moderately viscous fluid, which is easily manipulated, poured and injected. However, when
this hydrogel is exposed to physiologic temperature and pH (i.e., approximately 37°C and pH
7.4) a cubic phase (i.e., a liquid crystalline gel) results consisting of approximately 15% -
40% H2O and 85% - 60% GMO and is expected to have an equilibrium water content (i.e.,
maximum water content in the presence of excess water) of approximately 35% - 40% by
weight. This cubic phase is highly viscous and will exceed 1.2 million centipoise (cp).
Example IV
A Thermoreversible Gel
This example demonstrates that sirolimus may be incorporated into a thermoreversible
gel polymer composition having internal micellular components sufficient for controlled
release.
This polymer composition is represented by the composition trademarked as Flogel®
(Alliance Pharmaceuticals; San Diego, CA) and comprises a polyoxyethylene-
polyoxypropylene block copolymer having the formula HO(C2 H4 O)b (C3 H6 O\ (C2 H4 O)b
H, wherein a is an integer such that the hydrophobe base represented by (C3 Hg O\ has a
molecular weight of at least about 900, preferably at least about 2500, most preferably at least
about 4000 average molecular weight, as determined by hydroxyl number. Similar polymer
compositions may also be produced having a polyoxyproplyene hydrophobe base average
molecular weight of about 4000, a total average molecular weight of about 12,000 and
containing oxyethylene groups in the amount of about 70% by weight of the total weight of
the copolymer. A preferred copolymer is a tri-block copolymer containing two
polyoxyethylene blocks flanking a central polyoxypropylene block and is sold under the
trademark Pluronic® F-127 (BASF Corp, Parsippany, N.J.).
In this example, Pluronic® F-127 mixed with sirolimus, as discussed herein, is placed
in water, and the Pluronic® F-127 self-assembles so as to remove contact between the
polyoxypropylene groups and water (i.e. self-assembly is driven by a hydrophobic effect).
These self-assembled units are termed micelles within which are trapped sirolimus drug

molecules. The structure of the micelles and the interactions between them is strongly
dependent on temperature. A large increase in solution viscosity (i.e. gel-phase formation) is
noted with increasing temperature due to the organization of the micelles into a
three-dimensional cubic array (see Example HI). This gelation time may be controlled by the
addition of a modifying polymer including, but not limited to, cellulose derivatives.
The Pluronic* F-127-sirolimus solution is maintained at + 4°C until the time of use.
When the chilled solution is placed on or within a living tissue the solution will gel to form a
solid matrix on the surface of the tissue. During the subsequent controlled dissolution of the
matrix, the sirolhnus will be slowly released into the immediate environment to prevent scar
tissue and adhesion formation. The dissolution rates of thermoreversible gels may be
controlled by compounds including, but not limited to, fatty acid soap derivatives. It is
expected that the gelled matrix begins dissolution during the first day after administration and
is completely dissolved following twenty-one days after administration.
Example V
A Fibrin-Based Microparticle Bioadhesive
This example described the preparation of a powdered fibrin bioadhesive containing a
sirolimus compound. Specifically, the composition comprises microparticles containing a
fibrinogen-thrombin matrix and sirolimus. This protocol entails the preparation of two
separate powders (i.e, a fibrinogen powder and a thrombin powder) that are mixed together
just prior to use. United States Patent No. 6,113,948 To Heath et al. (herein incorporated by
reference).
Briefly, the first powder comprises fibrinogen and sucrose and the second powder
comprises thrombin, CaCl2, sirolimus and mannitol. Fibrinogen is first formulated with 600
mg sucrose. The resulting composition is then spray-dried using a Mini Spray Dryer with a
collecting vessel under the following conditions:



A 20% final excipient loading is expected along with a fibrinogen theoretical activity
of 10 mg/100 mg. This indicates a Ml retention of the fibrinogen bioactivity.
The second powder is prepared by dissolving 1 g D-mannitol in 10 ml of 40 mM
CaCl2 with any soluble form of sirolimus, as described herein, at a concentration sufficient to
obtain a final concentration of 2 mg/15 cm2 of tissue surface. The resultant solution is used
to reconstitute 1 vial of thrombin. The spray-drying conditions are essentially the same as for
the first powder, except that the outlet temperature is 62°C, and the feed rate is reduced to
0.75 g/min.
A thrombin clotting assay should reveal a thrombin activity of 91.86 units/100 mg that
will compared favorably with the theoretical activity, of 93 units/100 mg. This indicates full
retention of thrombin bioactivity.
The first and second microparticle powders are then mixed to form a 50:50 blend in a
glass vial by placement on a roller mixer for 20 minutes. This activated mixture is then
applied to a biological tissue.
Example VI
A Dual Component Bioadhesive With PLGA Microspheres
This example describes a composition for a sirolimus-eluting bioadhesive consisting of
proteinaceous materials and a cross-Unking agent.
Dry plasma solids are obtained by lyophilizing fresh frozen human plasma.
Thereafter, water is added to this solid to produce a viscous solution containing 45% of solids
by weight to create Solution A. Sirolimus-eluting microspheres, prepared in accordance with
Example I, are then added to Solution A. Solution B is prepared by creating an aqueous 10%
(w/w) glutaraldehyde mixture. The bioadhesive properties may be tested by lightly spraying
two rectangular (i.e., 2.5 cm x 2.5 cm) blocks of meat with Solution B on the surfaces to be
bonded. The surfaces are then coated with Solution A to a thickness of 1-2 mm, and again

sprayed with Solution B. This process will result in a ratio of Solution A to Solution B of 7
to 1 by weight. The surfaces are then joined within about 10 seconds of the application of
Solution A and held in position until cure was complete, generally 15-60 seconds, depending
on temperature and on the effectiveness of mixing Solution A and Solution B.
If the sequence of application of Solution A and Solution B is reversed or if Solution
A and Solution B are applied simultaneously or if Solution A and Solution B are pre-mixed
immediately prior to application, essentially the same bond strengths are expected to be
observed. Sirolimus elution may be tested by placing the sample that includes the bioadhesive
layers into a glass vial filled with 25 ml phosphate buffered isotonic saline (PBS: pH 7.4;
37°C). At predetermined intervals the buffer solution may be removed and the is vial refilled
with fresh PBS. The sirolimus in the removed PBS is then extracted by mixing with 1:1
chloroform. The chloroform is separated and filtered through a polyethylene Frit and
YLON+GL0.45 jim filter (Millipore). The released amount of sirolimus may be determined
in triplicate by UV spectroscopy at 280 nm and compared to a standard calibration curve.
Example VII
A Foam Cream
This example describes a composition for a pharmaceutical foam cream containing
sirolimus.
The cream is produced by combining the following ingredients in a turbo diffuser:
sirolimus 1%; white vaseline 12%; liquid paraffin 74%; white wax 3%; hydrogenated castor
oil 5%; and methylglucose dioleate 5%. The operation of the turbo diffuser will first melt
together the vaseline, paraffin, glucose and wax components by warming to a temperature of
72QC while slowly stirring. Then hydrogenated castor oil is added to the mixture, which is
then homogenized with a central turbo homogenizer. After cooling to room temperature,
sirolimus is added to the mixture and then homogenized with the turbo diffuser under a light
vacuum of 500 mm of mercury. The resulting cream is filled into suitable containers.

Example VHI
A Foam Cream Canister
This example describes the filling requirements and composition for a sirolimus foam
cream application canister.

In a first stainless steel container having an external jacket for warming, and a stirring
blade, 3.2 kg cetyl stearyl alcohol (USP) is melted in 43.8 kg mineral oil (USP) to a
temperature of 65° ± 0.5°C while stirring. In a second stainless steel turbo vacuum diffuser
provided with a water jacket for heating and cooling, a stirring blade, scraper and central
turbo homogenizer, 29 kg mineral oil (USP) and 4 kg sirolimus foam cream made in
accordance with Example VII are placed together. These two components are mixed by
stirring at a low rate for 30 minutes under a light vacuum (500 mmHg). Thereafter, the
above cetyl stearyl alcohol in mineral oil solution is cooled to 45°C and added to the
sirolimus/mineral oil mixture with continuous stirring under light vacuum for an additional 10
minutes while cooling the mixture to room temperature. The mixture is then subdivided by
means of a filling machine into approximately 20,000 canisters. The canisters are thereafter
closed with a polyethylene valve and for filling with propellant gas Purifair™ 3.2 and a
polyethylene tube is inserted in the valve to facilitate complete delivery when the valve is
depressed.
Example IX
An Elastomeric Foam
This example describes the production of a sirolimus foam scaffolding composition.
A random copolymer of e-caprolactone-glycolide (PCL/PLGA) with a 35/65 molar
composition is synthesized by a ring-opening polymerization reaction. Bezwada et ai.

*fclastomeric Medical Device. U.S. Pat. No. 5,468,253 (herein incorporated by reference). A
diethylene glycol initiator is added and is adjusted to a concentration of 1.15 mmole/mole of
monomer to obtain a dried polymer having the following characteristics: i) an inherent
copolymer viscosity of 1.59 dL/g in hexafluoroisopropanol at 25°C; ii) a PCL/PGA molar
ratio of 35.5/64.5 by proton NMR with about 0.5% residual monomer; iii) a glass transition
and melting point of approximately -10°C and 65°C, respectively.
A 5% (w/w) 35/65 PCL/PGA polymer/1,4-dioxane solution containing a desired
concentration of sirolimus is next prepared by gentle heating to 60 ± 0.5°C and continuously
stirring for at least 4 hours but not more man 8 hours. The solution is prepared in a flask
with a magnetic stir bar. A clear homogeneous solution is then obtained by filtering the
solution through an extra coarse porosity filter (i.e., a Pyrex brand extraction thimble with
fritted disc) using dry nitrogen.
The solution is thereafter lyophilized, using for example, a laboratory scale
lyophilizer-Freezemobile 6 (Virtis™). The freeze-dryer is preset at 20°C under a dry nitrogen
atmosphere and allowed to equilibrate approximately 30 minutes. The PCL/PGA polymer
solution is poured into the molds just before the actual start of the cycle. A glass mold is
preferred but a mold made of any material that is: i) inert to 1,4-dioxane; ii) has good heat
transfer characteristics; and iii) has a surface that enables the easy removal of the foam. The
best results are expected with a glass mold or dish weighing 620 grams, having optical glass
5.5 mm thick, and being cylindrical with a 21 cm outer diameter and a 19.5 cm inner
diameter. Next the following steps are followed in a sequence to make 2 mm thick foam
pieces:
i) The glass dish with the solution is carefully placed (without tilting) on the shelf
of the lyophilizer, which is maintained at 20°C. The cycle is started and the
shelf temperature is held at 20°C for 30 minutes for thermal conditioning.
ii) The solution is then cooled to -5°C by cooling the shelf to -5°C.
iii) After 60 minutes of freezing at -5°C, a vacuum is applied to initiate primary
drying of the dioxane by sublimation; approximately one hour of primary
drying under vacuum at r5°C is needed to remove most of the solvent. At the
end of this drying stage the vacuum level will typically reach about 50 mTorr

or less.
iv) Next, secondary drying under a 50 mTorr vacuum or less is performed in two
stages to remove the adsorbed dioxane. In the first stage, the shelf temperature
is raised to + 5°C for approximately 1 hour. In the second stage, temperature
is raised to 20°C for approximately 1 hour.
v) At the end of the second stage, the lyophilizer is brought to room temperature
and the vacuum is broken with nitrogen. The chamber is then purged with dry
nitrogen for approximately 30 minutes before opening the door.
As one skilled in the art would know, the conditions described herein are
typical and operating ranges depend on several factors e.g.: concentration of the solution;
polymer molecular weights and compositions; volume of the solution; mold parameters;
machine variables like cooling rate, heating rates; and the like. The above described process
is expected to result in elastomeric foams having a random microstructure.
Example X
Spray Application By A Catheter
This example provides a method and a device to administer sirolimus in an appropriate
vehicle, as described herein, as a spray during an endoscopic procedure using an
accompanying catheter.
A "side hole catheter" has tiny round side holes cut into the catheter near a closed
distal end. (e.g., Figure 11) This catheter is constructed of a flexible, elongated,
biocompatible polymer tubing which is hollow and thin-walled and should have a uniform
diameter of 2 to 20 French, but preferably 5 to 10 French. Radioopaque markings on the
catheter allows easy tracking of the catheter position via fluoroscopy. The catheter contains a
medium comprising sirolimus. In practice, the catheter is inserted into a lumen of an
endoscope system in accordance with standard approved procedures, and is moved carefully
such that distal end of the catheter is positioned into or near the application site. A
pharmaceutical solution of sirolimus is then injected under gentle pressure from a syringe-like
reservoir attached to a female Luer lock and is impelled toward distal end of the catheter,
emerging through the side holes and onto the application site. Alternately, a spray can or

other apparatus under pressure may be attached to the Luer lock and spray administered via
the side holes.
Alternatively, a "slit catheter" (Figure 12), also composed of a flexible catheter 90
comprising a hollow, thin-walled, biocompatible polymer material 92 into which extremely
thin slits 95 that are laser cut at regular intervals near a closed distal end 97. These slits are
tight enough that infusate will not escape unless the fluid pressure within the catheter reaches
a critical point that cause the slits to distend simultaneously and temporarily open. This
catheter also contains exterior radiopaque markers to assist in the positioning of the device.
These slit catheters are also used in conjunction with an automated, piston-driven,
pulsed infusion devices that are capable of delivering low volume regulated pulses of drug
infusion at the proximal end of the catheter. When a pulse is delivered, the pressure within
the catheter rises momentarily thus causing the slits to open momentarily to administer the
sirolimus. Slit catheters are preferable to side hole catheters since, in the former type, the
spray is delivered uniformly through all slits along the entire length of the catheter, whereas
sprays from a "side hole" catheter are administered mainly from the most proximal side holes.
Example XI
Spray Application By A Single Dose Dispenser
This example describes a single dose spray dispenser that is capable of applying a
single dose of sirolimus, tacrolimus and analogs of sirolimus. It is understood by one skilled
in the art that the basic concept described below may be modified and adapted to administer a
single dose either internally or externally. For example, the dispenser described below may
be reconfigured for operation with a catheter for administration to an intraluminal site within
the body. Depending upon the size of the surgical site, a medical practitioner may dispense
one or more cans at any one particular site.
The Dispenser Device
A dispenser device for spraying a single dose of sirolimus intended to cover about 50
cm2 of wound at the rate of about 200 jig/cm2 will have a cylinder containing a predetermined
dose of a liquid medium containing sirolimus, tacrolimus or an analog of sirolimus. A piston
will slide in a sealed manner within the cylinder between a storage position in which it

isolates the cylinder to an actuated position. An outlet passage will connect the cylinder to an
outlet orifice where the entire single dose of liquid is expelled from the device when the
piston is slid from the storage to the actuated position. Martin et al, Device For Dispensing
A Single Dose Of Fluid. United States Patent No. 6,345,737 (herein incorporated by
reference).
The Liquid Medium
Sirolimus will be dissolved in olive oil at a concentration of 1 mg/ml. Alternatively,
soluble monoacyl and diacyl derivatives of sirolimus are prepared according to known
methods. Rakhit, U.S. Pat. No. 4,316,885 (herein incorporated by reference). These
derivatives are used in the form of a sterile solution or suspension containing other solutes or
suspending agents, for example, enough saline or glucose to make the solution isotonic, bile
salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitol and its
anhydrides copolymerized with ethylene oxide) and the like. Furthermore, water soluble
prodrugs of sirolimus may be used including, but not limited to, glycinates, propionates and
pyrrolidinobutyrates. Stella et al., U.S. Pat. No. 4,650,803 (herein incorporated by reference).
Controlled Release
Alternatively, the liquid media described above is prepared using microspheres
prepared according to Example I or using liposomes prepared according to Example III.
Example XII
Aerosolizaton
This example describes one method of providing a sirolimus aerosol spray to an area
of interest.
The Nebulizer
A nebulizer will transform solutions or suspensions of sirolimus according to any of
the applicable Examples discusses herein, into a therapeutic aerosol mist either by means of
acceleration of a compressed gas, typically air or oxygen, through a narrow venturi orifice. In
particular, embodiments of sirolimus media exhibiting controlled release capabilities are
preferred. Sirolimus is present in a liquid carrier in an amount of up to 5% w/w, but
preferably less than 1% w/w of the formulation. The carrier is typically water or a dilute

aqueous alcoholic solution, preferably made isotonic with body fluids by the addition of, for
example, sodium chloride. Solubility enhancing agents are well known in the art and may be
added as deemed required depending upon the required concentration. Optional additives
include preservatives if the formulation is not prepared sterile, for example, methyl
hydroxybenzoate, antioxidants, volatile oils, buffering agents and surfactants.
The present invention contemplates the use of many devices to generate an aerosol and
the following exemplary device is not intended to limit the invention. The nebulizer device
has a lever that activates an air spring-valve joint directly connected to an external source of
pressurized air or other gaseous propellant such that the air enters an air chamber. An air
channel will extend from an air chamber to the distal end of the aerosolization apparatus. The
air channel terminates into a rod extension that contains the aperture aerosolization tips.
A fluid chamber tip also include apertures that communicate with the air channels.
When the sirolimus fluid chamber tip end is inserted into the air channel aperture, no air
passes out of air chamber. When the fluid chamber tip end is, however, withdrawn from the
air channel aperture, air and fluid mix into an aerosol and exit the apparatus through a
dispensing tip.
The Liquid Medium
Sirolimus will be dissolved in olive oil at a concentration of at least 10 mg/ml.
Alternatively, soluble monoacyl and diacyl derivatives of sirolimus are prepared according to
known methods. Rakhit, U.S. Pat. No. 4,316,885 (herein incorporated by reference). These
derivatives are used in the form of a sterile solution or suspension containing other solutes or
suspending agents, for example, enough saline or glucose to make the solution isotonic, bile
salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitol and its
anhydrides copolymerized with ethylene oxide) and the like. Furthermore, water soluble
prodrugs of sirolimus may be used including, but not limited to, glycinates, propionates and
pyrrolidinobutyrates. Stella et al, U.S. Pat. No. 4,650,803 (herein incorporated by reference).

Example XIII
A Multiple Lumen Catheter
This example describes a catheter capable of co-administration of several sirolimus
solutions simultaneously, or mixing a sirolimus solution with a non-sirolimus solution into a
single composition. Specifically contemplated is the mixing of two separate components in
order to spray a sirolimus-containing bioadhesive.
A device for applying two-component products, such as medical tissue bioadhesive,
has a flat head piece connected at the front end to a tubular body. A multiple lumen tube is
therewith expected to be in communication with a tubular body. The dorsal surface of the
head piece also has portions of two cannula hubs. A multiple lumen tube is comprised of
three lumina which extend in parallel from the inner end of the lumen tube to the discharge
end. Two lumina are connected to each of two syringes (respectively), either barrels of which
may contain a sirolimus-containing composition. The plunger rods of the syringes are
coupled by a bridging member such that both are operated simultaneously to permit equal
mixing and administration of the compositions in both barrels.
Two of the cannula hubs, partially included in the head piece, are connected to rigid
cannulas preferably made of metal. The two metal cannulas are oriented in the head piece
such that they extend in V-shape. A third lumen is an end of a connecting tubule and is
connected to a soft flexible air tube. An air tube also extends from the tip of the V formed
by the two metal cannulas straight to the rear end of the head piece.
The air tube is in direct communication with the third lumen of the multiple lumen
tube through the connecting tubule. The precise flow of the compositions from the two
syringe barrels and the air flow is expected to emerge from the catheter close together as a
thin jet from the discharge end of the multiple lumen tube. The compositions from the two
syringe barrels are sprayed in an optimal mixture by the air flow so that the treated site is
supplied with a sufficient quantity of dispersed sirolimus bioadhesive. Due to the separate
transport of the compositions from the two syringe barrels and the air in different lumens, the
compound containing material is only mixed when past the discharge end of the multiple
lumen tube. Accordingly, the portions of the compound containing material from the two
syringe barrels are dosed exactly and the composition of a sirolimus bioadhesive is always

Example XIV
Bioadhesive Applicator Device
This example describes one embodiment of a bioadhesive applicator device (see Figure
13).
The applicator is constructed as a pair of syringes 105 and 106, each of which has
plungers 101 and 102 which variably slide within a hollow of each respective syringe body
between a fully retracted position to a fully compressed position. Each of the syringes 105
and 106, respectively, contain a different material {i.e., for example, thrombin versus fibrin)
that, become an adhesive compound when mixed. The syringes merge into a common mixing
area 120 at one end, wherein the mixing area 120 is adapted to connect with each outlet of
syringes 105 and 106. The plungers 101 and 102 will push the respective medium out of
each syringe 105 and 106, whereupon mixing occurs prior to exiting from a nozzle 122 as a
single stream. After the mixed adhesive medium exits the applicator, the mixture will harden
into a bioadhesive onto the target tissue site.

Claims
We claim:
1. A drug attached to a carrier, the drug being selected from the group consisting of
sirolimus, tacrolimus, everolimus and the analogs and derivatives thereof, the carrier
onto which the drug is attached being selected from the group consisting of
microparticles, gels, xerogels, bioadhesives, foams and liquids.
2. The drug attached to a carrier-ef Claim 1, wherein the carrier comprises a
biocompatible material.
3. The drug attached to a carrier ef Claim 1, wherein the carrier comprises a
biodegradable material.
4. The drug attached to a carrier of Claim 1, wherein the microparticles are selected from
the group consisting of microspheres, microencapsulating particles, microcapsules and
liposomes.
5. The microparticle ei Claim 4 comprising a polymer selected from the group consisting
of poly(lactide-co-glycolide), aliphatic polyesters, poly-glycolic acid, poly-lactic acid,
hyaluronic acid, modified polysacchrides, poly(ethylene oxide), lecithin and
phospholipids.
6. The drug attached to a carrier ofr Claim 1, wherein the carrier comprises a material
selected from the group consisting of poly(lactide-co-glycolide), aliphatic polyesters,
poly-glycolic acid, poly-lactic acid, hyaluronic acid, modified polysacchrides,
poly(ethylene oxide), lecithin, phospholipids, fibrin sealants, polyethylene oxide,
polypropylene oxide, block polymers of polyethylene oxide and polypropylene oxide,
polyethylene glycol, methacrylates and cyanoacrylates.

The drag attached to a carrier^CIaim 1, wherein the carrier releases said drug in a
controlled release manner.
8. The drug attached to a carrier^? Claim 1, wherein the carrier is colored.
9. A medium, comprising a compound selected from the group consisting of sirolimus,
tacrolimus, analogs of sirolimus and phannaceutically acceptable salts thereof, wherein
said medium is selected from the group consisting of microparticles, gels, xerogels,
bioadhesives and foams.
10. The mediuny*/Claim 9, wherein said medium comprises a biocompatible material.
11. The medium of Claim 9, wherein said medium comprises a biodegradable material.
A
12. The medium ^Claim 9, wherein said microparticles are selected from the group
consisting of microspheres, microencapsulating particles, microcapsules and liposomes.
13. The mediumyf Claim 9, wherein said medium is colored.
14. The medium/f Claim 9, wherein said analog of sirolimus is selected from the group
consisting of everolimus, CCI-779, ABT-578, 7-epi-rapamycin, 7-thiomethyl-
rapamycin, 7-epi-trimethoxyphenyl-rapamycin, 7-epi-thiomethyl-rapamycin,
7-demethoxy-rapamycin, 32-demethoxy-rapamycin and 2-desmethyl-rapamycin.
15. The medium pf Claim 9, further comprising a second compound selected from the
group consisting of antiinflammatory, corticosteriods, antithrombotics, antibiotics,
antivirals, analgesics and anesthetics.
16 A device said device comprising a reservoir comprising the medium of Claim 9 and
capable of delivering said medium of Claim 9 to a surgical site.

17. The device^ Claim 16, wherein said delivering is in the form of a
spray.
18. The device of Claim 16, wherein said delivering is in the form of an
aerosol.
19. The device ofClaim 16, wherein said device comprises a catheter.
r
20. The device g£ Claim 16, wherein said device is configured for
endoscopic surgery.
21. The device pt Qaim 16, wherein said device is configured for
fluoroscopic surgery.
22. A medical device wherein at least a portion of said device is coated
with the medium of Claim 9.
23. A collection of microspheres comprising a biocompatible material for
placement at or near the site of a surgical procedure to reduce the
formation of scar tissue and adhesions, the microspheres having a
diameter between 0.1 and 100 microns and a cytostatic and
antiproliferative drug attached to the microspheres that is adapted for
release over time.
24. A method for delivering a gel or liquid comprising microparticles
having an attached compound selected from the group consisting of
sirolimus, tacrolimus and analogs of sirolimus to a surgical site.

25. A gel or liquid, comprising at least one compound selected from the
group consisting of sirolimus, tacrolimus, analogs of sirolimus and
pharmaceutically acceptable salts thereof, wherein said compound is
attached to a microparticle.

A drug attached to a carrier, the drug being selected from the group consisting of
sirolimus, tacrolimus, everolimus and the analogs and derivatives thereof, the
carrier onto which the drug is attached being selected from the group consisting
of microparticles, gels, xerogels, bioadhesives, foams and liquids.

Documents:

02429-kolnp-2005-abstract.pdf

02429-kolnp-2005-claims.pdf

02429-kolnp-2005-description complete.pdf

02429-kolnp-2005-drawings.pdf

02429-kolnp-2005-form 1.pdf

02429-kolnp-2005-form 2.pdf

02429-kolnp-2005-form 3.pdf

02429-kolnp-2005-form 5.pdf

02429-kolnp-2005-international publication.pdf

2429-kolnp-2005-granted-abstract.pdf

2429-kolnp-2005-granted-assignment.pdf

2429-kolnp-2005-granted-claims.pdf

2429-kolnp-2005-granted-correspondence.pdf

2429-kolnp-2005-granted-description (complete).pdf

2429-kolnp-2005-granted-drawings.pdf

2429-kolnp-2005-granted-examination report.pdf

2429-kolnp-2005-granted-form 1.pdf

2429-kolnp-2005-granted-form 18.pdf

2429-kolnp-2005-granted-form 2.pdf

2429-kolnp-2005-granted-form 3.pdf

2429-kolnp-2005-granted-form 5.pdf

2429-kolnp-2005-granted-reply to examination report.pdf

2429-kolnp-2005-granted-specification.pdf


Patent Number 235982
Indian Patent Application Number 2429/KOLNP/2005
PG Journal Number 37/2009
Publication Date 11-Sep-2009
Grant Date 10-Sep-2009
Date of Filing 30-Nov-2005
Name of Patentee AFMEDICA, INC.
Applicant Address 259 EAST MICHIGAN AVENUE, SUITE 409, KALAMAZOO, MI
Inventors:
# Inventor's Name Inventor's Address
1 FISCHELL, SARAH, T. 71 RIVERLAWN DRIVE, FAIRHAVEN, NJ 07704
2 FISCHELL, ROBERT, E. 14600 VIBURNUM DRIVE, DAYTON, MD 21036
3 WALDORF, CLAYTON, MACKENZIE 10844 SOUTH INTERLAKEN DRIVE, RICHLAND, MI 49083
4 FISCHELL, TIM, A. 1701 EMBURY ROAD, KALAMAZOO, MI 49008
5 WALDORF, CLAYTON, MACKENZIE 10844 SOUTH INTERLAKEN DRIVE, RICHLAND, MI 49083
6 FISCHELL, ROBERT, E. 14600 VIBURNUM DRIVE, DAYTON, MD 21036
7 FISCHELL, SARAH, T. 71 RIVERLAWN DRIVE, FAIRHAVEN, NJ 07704
8 FISCHELL, TIM, A. 1701 EMBURY ROAD, KALAMAZOO, MI 49008
PCT International Classification Number A61K
PCT International Application Number PCT/US2004/014118
PCT International Filing date 2004-05-06
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 10/431,701 2003-05-07 U.S.A.