U.S. patent application number 10/934067 was filed with the patent office on 2005-06-30 for implantable system with drug-eluting cells for on-demand local drug delivery.
This patent application is currently assigned to Medtronic, Inc.. Invention is credited to Donovan, Maura G., Soykan, Orhan.
Application Number | 20050143802 10/934067 |
Document ID | / |
Family ID | 22508514 |
Filed Date | 2005-06-30 |
United States Patent
Application |
20050143802 |
Kind Code |
A1 |
Soykan, Orhan ; et
al. |
June 30, 2005 |
Implantable system with drug-eluting cells for on-demand local drug
delivery
Abstract
An implantable system that includes a carrier and eukaryotic
cells, which produce and release a therapeutic agent, and a
stimulating element for stimulating the release of the therapeutic
agent. The system can also include a sensing element for monitoring
a physiological condition and triggering the stimulating element to
stimulate the delivery device to release the therapeutic agent.
Alternatively, the patient in which the system is implanted can
activate the stimulating element to release the therapeutic
agent.
Inventors: |
Soykan, Orhan; (New
Brighton, MN) ; Donovan, Maura G.; (St. Paul,
MN) |
Correspondence
Address: |
Kenneth J. Collier
Medtronic, Inc.
710 Medtronic Parkway N.E.
Minneapolis
MN
55432
US
|
Assignee: |
Medtronic, Inc.
|
Family ID: |
22508514 |
Appl. No.: |
10/934067 |
Filed: |
September 3, 2004 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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10934067 |
Sep 3, 2004 |
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09745144 |
Dec 20, 2000 |
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6824561 |
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10934067 |
Sep 3, 2004 |
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09144420 |
Aug 31, 1998 |
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6206914 |
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10934067 |
Sep 3, 2004 |
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09070480 |
Apr 30, 1998 |
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Current U.S.
Class: |
623/1.11 |
Current CPC
Class: |
A61L 2300/414 20130101;
A61L 2300/602 20130101; A61L 29/16 20130101; A61L 31/005 20130101;
A61L 27/54 20130101; A61L 27/3843 20130101; A61F 2/06 20130101;
C12N 5/069 20130101; A61F 2250/0067 20130101; A61L 31/10 20130101;
A61F 2250/0001 20130101; A61N 1/36542 20130101; A61L 2300/64
20130101; A61K 38/49 20130101; A61K 9/0024 20130101; A61L 31/16
20130101; A61K 48/00 20130101; A61F 2/02 20130101; A61L 29/005
20130101; A61L 27/3804 20130101; A61L 2300/252 20130101; A61L
2300/22 20130101; A61L 27/3808 20130101; A61K 2035/126 20130101;
A61K 9/0009 20130101; A61L 2300/00 20130101; A61F 2/88 20130101;
A61L 2300/416 20130101; A61L 2300/42 20130101; A61K 38/4866
20130101; A61F 2/82 20130101; A61L 2300/254 20130101 |
Class at
Publication: |
623/001.11 |
International
Class: |
A61F 002/06 |
Claims
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41. An implantable system comprising: (a) a delivery device
comprising a carrier and eukaryotic cells that produce at least one
therapeutic agent; and (b) a stimulating element for stimulating
the release of the therapeutic agent from the delivery device;
wherein the stimulating element can be activated by a patient in
which it is implanted on demand.
42. The implantable system of claim 41 wherein the delivery device
comprises: (a) an intraluminal stent comprising: (i) a lumen-wall
contacting surface; (ii) a lumen-exposed surface; (iii) a first
polymer composition covering at least a portion of the stent; and
(b) endothelial cells associated with the first polymer
composition; wherein the endothelial cells produce at least one
therapeutic agent.
43. A method of local delivery of a therapeutic agent to a patient,
the method comprising: (a) providing an implantable system
comprising: (i) a delivery device comprising a carrier and
eukaryotic cells that produce at least one therapeutic agent; and
(ii) a stimulating element for stimulating the release of the
therapeutic agent from the delivery device; and (b) implanting the
implantable system into the body of a patient; wherein the
therapeutic agent is released from the delivery device when the
patient detects a change in a physiological condition and
communicates with the electrical stimulating element to trigger the
release of the therapeutic agent.
44. The method of claim 43 wherein the delivery device comprises:
(a) an intraluminal stent comprising: (i) a lumen-wall contacting
surface; (ii) a lumen-exposed surface; (iii) a first polymer
composition covering at least a portion of the stent; and (b)
endothelial cells associated with the first polymer composition;
wherein the endothelial cells produce at least one therapeutic
agent.
45. The method of claim 43 wherein the patient activates the
stimulating element using radio frequency, infrared, or acoustic
pulses.
Description
[0001] This application is a Continuation-In-Part of U.S. patent
application Ser. No. 09/070,480 filed 30 Apr. 1998, which is
incorporated by reference.
FIELD OF THE INVENTION
[0002] The present invention relates to implantable systems that
include medical devices (e.g., stents, vascular grafts, stent
grafts) that function as a carrier for eukaryotic cells (e.g.,
genetically engineered endothelial cells). Such cells are capable
of producing and releasing a therapeutic agent (e.g., tissue-type
Plasminogen Activator) for on-demand localized treatment of
conditions such as coronary artery disease. The cells or the device
carrying them release the therapeutic agent upon the application of
a stimulus (e.g., electrical stimulus).
BACKGROUND OF THE INVENTION
[0003] Coronary Artery Disease (CAD) affects 1.5 million people in
the USA annually. About 10% of these patients die within the first
year and the rest suffer from myocardial infarction and develop
related symptoms, such as arrhythmias, CHF, and mechanical
complications (e.g., aneurysm, thrombus formation, pericarditis).
During CAD, formation of plaques under the endothelial tissue
narrows the lumen of the coronary artery and increases its
resistance to blood flow, thereby reducing the O.sub.2 supply.
Injury to the myocardium (i.e., the middle and thickest layer of
the heart wall, composed of cardiac muscle) fed by the coronary
artery begins to become irreversible within 0.5-1.5 hours and is
complete after 6-12 hours, resulting in a condition called
myocardial infarction. If the ischemia due to stenosis of the
coronary artery lumen could be reduced by increasing the blood
circulation to the myocardium, the major cause of most of the heart
disease would be eliminated.
[0004] Current and proposed treatments for coronary artery disease
typically focus on pharmacological approaches and surgical
intervention. For example, angioplasty, with and without stents, is
a well known technique for reducing stenosis. Systemically
administered drugs (e.g., anticoagulants) are also commonly used,
however, such drugs become diluted, which can reduce their potency
by the time they reach the remote site. Furthermore, systemic
administration can be deleterious because it can lead to
complications as a result of the high dosages required upon
administration to allow for dilution that occurs during transport
to the remote site. Therefore, localized delivery of therapeutic
agents is preferred. Local delivery is advantageous in that the
effective local concentration of delivered drug is much higher than
can normally be achieved by systemic administration.
[0005] Stents have been used as delivery vehicles for therapeutic
agents (i.e., drugs). Intravascular stents are generally
permanently implanted in coronary or peripheral vessels. Stent
designs include those of U.S. Pat. No. 4,733,655 (Palmaz), U.S.
Pat. No. 4,800,882 (Gianturco), or U.S. Pat. No. 4,886,062
(Wiktor). Such designs include both metal and polymeric stents, as
well as self-expanding and balloon-expandable stents. Stents are
also used to deliver a drug (e.g., antiplatelet agents,
anticoagulant agents, antimicrobial agents, antimetabolic agents)
at the site of contact with the vasculature, as disclosed in U.S.
Pat. No. 5,102,417 (Palmaz), and in International Patent
Application Nos. WO 91/12779 (Medtronic, Inc.) and WO 90/13332
(Cedars-Sanai Medical Center), for example. Anticoagulant
substances such as heparin and thrombolytic agents have also been
incorporated into a stent, as disclosed, for example, in U.S. Pat.
No. 5,419,760 (Narciso, Jr.) and U.S. Pat. No. 5,429,634 (Narciso,
Jr.). Stents have also been used to deliver viruses to the wall of
a lumen for gene delivery, as disclosed in U.S. patent application
Ser. No. 08/746,404, filed Nov. 8, 1996 (Donovan et al.).
[0006] Stents seeded with autologous endothelial cells (Dichek, et
al., Circulation, 80,1347-1353 (1989)) are disclosed as a method
for delivering recombinant protein over time to the vascular wall.
The concentration of secreted protein produced by the endothelial
cells that is required to treat the surrounding vascular tissue can
be significantly higher than could be tolerated if delivered
systemically. However, in order to be effective, it is not only
necessary to release a high dose of the drug, but it is also
necessary to achieve controlled release, e.g., immediately after an
occlusion. Thus, it would be desirable to control the release of
such cellular components into the surrounding tissue when needed
(i.e., on demand). The present invention provides such a
system.
[0007] Many of the following lists of patents and nonpatent
documents disclose information related to the local delivery of
therapeutic agents using medical devices, such as stents. Others
are directed toward stent designs and other medical devices as well
as genetically engineered cells, for example.
1TABLE 1a Patents Patent No. Inventor(s) 3,523,807 Gerendas
4,476,868 Thompson 4,540,573 Neurath et al. 4,548,736 Muller et al.
4,556,063 Thompson et al. 4,733,655 Palmaz 4,800,882 Gianturco
4,821,723 Baker et al. 4,886,062 Wiktor 4,944,659 Labbe et al.
5,102,417 Palmaz 5,131,388 Pless 5,144,949 Olson 5,158,078 Bennett
et al. 5,192,297 Hull 5,199,428 Obel et al. 5,207,218 Carpentier et
al. 5,312,453 Shelton et al. 5,314,430 Bardy 5,330,507 Schwartz
5,354,316 Keimel 5,372,600 Beyer et al. 5,409,009 Olson 5,419,760
Narciso, Jr. 5,429,634 Narciso, Jr. 5,510,077 Dinh et al. 5,545,186
Olson et al. 5,674,722 Mulligan et al. 5,702,427 Ecker et al. U.S.
Pat. Appl. Donovan et al. 08/746,404, filed Nov. 8, 1996
[0008]
2TABLE 1b Foreign Patent Documents Document No. Applicant
Publication Date WO 90/13332 Cedars-Sanai Medical Ctr. Nov 15, 1990
WO 91/12779 Medtronic, Inc. Sep 5, 1991 WO 96/28841 Smela et al.
Sep 19, 1996 WO 96/34417 Smela et al. Oct 31, 1996
[0009]
3TABLE 1c Nonpatent Documents Bansal et al., "Calcium-Regulated
Secretion of Tissue Plasminogen Activator and Parathyroid Hormone
from Human Parathyroid Cells," J. Clin. Endocrin. Metab., 74,
266-271 (1992). "Bioartificial Polymeric Materials Obtained from
Blends of Synthetic Polymers with Fibrin and Collagen",
International Journal of Artificial Organs, 14(5) (1991). Blaese et
al., "T Lymphocyte-Directed Gene Therapy for ADA SCID: Initial
Trial Results After 4 Years," Science, 270, 475-480 (1995).
Bordignon et al., "Gene Therapy in Peripheral Blood Lymphocytes and
Bone Marrow for ADA Immunodeficient Patients," Science, 270,
470-475 (1995). Bouaziz et al., "Effect of constant and modulated
electrical charges applied to the culture material on PGI2 and TXA2
secretion by endothelial cells," Biomaterials, 16, 727-734 (1995).
Bouaziz et al., "Vascular endothelial cell responses to different
electrically charged poly(vinylidene fluoride) supports under
static and oscillating flow conditions," Biomaterials, 18, 107-112
(1997). Braunwald et al., "Unstable Angina: Diagnosis and
Management," Clinical Practical Guidelines, No. 10, U.S. Department
of Health and Human Services, Public Health Services, Agency for
Health Care Policy and Research, AHCPR Publication No. 94-0602,
27-92 (1994). Bugiardini et al., "Relation of Severity of Symptoms
to Transient Myocardial Ischemia and Prognosis in Unstable Angina,"
J. Am. Coll. Cardiol., 25, 597-604 (1995). Chowdhury et al.,
"Long-Term Improvement of Hypercholesterolemia After ex Vivo Gene
Therapy in LDLR-Deficient Rabbits," Science, 254, 1802-1804 (1991).
Dicheck et al., "Enhanced In Vivo Antithrombotic Effects of
Endothelial Cells Expressing Recombinant Plasminogen Activators
Transduced With Retroviral Vectors," Circulation, 93, 301-309
(1996). Dicheck et al., "Seeding of Intravascular Stents With
Genetically Engineered Endothelial Cells," Circulation, 80,
1347-1353 (1989). Dicheck et al., "Retroviral Vector-mediated Gene
Transfer into Endothelial Cells," Mol. Biol. Med., 8, 257-266
(1991). Dunn et al., "Seeding of Vascular Grafts With Genetically
Modified Endothelial Cells," Circulation, 93, 1439-1446 (1996).
Ekterae et al., "Retroviral vector-mediated transfer and expression
of human tissue plasminogen activator gene in human endothelial and
vascular smooth muscle cells," J. Vascular Surgery, 21, 953-962
(June 1995). Fisher et al., "Isolation and Characterization of the
Human Tissue-type Plasminogen Activator Structural Gene Including
Its 5' Flanking Region," J. Biol. Chem., 260, 11223-11230 (1985).
Gillis et al., "Secretion of Prostacyclin, Tissue Plasminogen
Activator and its Inhibitor by Cultured Adult Human Endothelial
Cells Grown on Different Matrices," Eur. J. Vasc. Endovasc. Surg.,
11, 127-133 (1996). Grossman et al., "Successful ex vivo gene
therapy directed to liver in a patient with familial
hypercholesterolaemia," Nature Genetics, 6, 335-341 (1994). Innis
et al. eds., "PCR Strategies", Academic Press, New York, New York
(1995). Kohn et al., "Engraftment of gene-modified umbilical cord
blood cells in neonates with adenosine deaminase deficiency,"
Nature Medicine, 1, 1017-1023 (1995). Levin et al., "Protein Kinase
C and the Stimulation of Tissue Plasminogen Activator Release from
Human Endothelial Cells," J. Biol. Chem., 264, 16030-16036 (1989).
Lilly, "Acute Myocardial Infarction", Pathophysiology of Heart
Disease, Lea & Febiger, Philadelphia, PA, Ch. 7, 113-129
(1993). Nabel et al., "Recombinant Gene Expression in Vivo Within
Endothelial Cells of the Arterial Wall," Science, 244, 1342-1344
(1989). Nilius et al., "Activation of a Cl.sup.- Current by
Hypotonic Volume Increase in Human Endothelial Cells," J. Gen.
Phys., 103 787-805 (1994). Ryan et al., "ACC/AHA Guidelines for the
Management of Patients With Acute Myocardial Infarction: A Report
of the American College of Cardiology/American Heart Association
Task Force on Practice Guidelines (Committee on Management of Acute
Myocardial Infarction)," J. Am. Coll. Cardiol., 28, 1328-1428
(1996). Sambrook et al, "Molecular Cloning: A Laboratory Manual",
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York,
New York (1989). Santell et al., "Cyclic AMP Potentiates Phorbol
Ester Stimulation of Tissue Plasminogen Activator Release and
Inhibits Secretion of Plasminogen Activator Inhibitor-1 from Human
Endothelial Cells," J. Biol. Chem., 263, 16802-16808 (1988). Smela
et al., "Controlled Folding of Micrometer-Size Structures,"
Science, 268, 1735-1738 (1995). Van den Eijnden-Schrauwen et al.,
"Studies on the Acute Release of Tissue-Type Plasminogen Activator
from Human Endothelial Cells In Vitro and in Rats In Vivo: Evidence
for a Dynamic Storage Pool," Blood, 85, 3510-3517 (1995). Williams
et al., "Introduction of new genetic material into pluripotent
haematopoietic stem cells of the mouse," Nature, 310, 476-480
(1984). Yanagisawa et al., "The Human Preproendothelin-1 Gene:
Possible Regulation by Endothelial Phosphoinositide Turnover
Signaling," J. Cardiovasc. Pharmacol., 13, S13-S17 (1989).
[0010] All patent and nonpatent documents listed in Table 1 are
hereby incorporated by reference herein in their respective
entireties. As those of ordinary skill in the art will appreciate
upon reading the Summary of the Invention, Detailed Description of
Preferred Embodiments, and Claims set forth below, many of the
systems, devices, and methods disclosed in these documents may be
modified advantageously by using the teachings of the present
invention.
SUMMARY OF THE INVENTION
[0011] The present invention provides implantable systems that
include delivery devices having a carrier and eukaryotic cells
associated therewith, optionally, within a containment vehicle.
Such cells are capable of producing at least one therapeutic agent
(i.e., drug), which is released from the delivery device upon the
application of a stimulus (e.g., electrical stimulus) for on-demand
(i.e., when needed) localized treatment of conditions such as
coronary artery disease or cerebral vascular occlusion, for
example. The cells are referred to herein as "drug-eluting"
cells.
[0012] Release of the therapeutic agent from the delivery device is
stimulated by a variety of methods, including electrical
stimulation, which can be used to create an electrical field or
mechanical stimulus, for example. This can be accomplished by
direct action on the cells, such as by stimulating the cellular
membrane to release the cellular products contained therein.
Alternatively, this can be accomplished by activating the cellular
products such that upon release they will function as therapeutic
agents. This can also be accomplished by stimulating a microscopic
containment vehicle that contains the cells, such as by stimulating
a synthetic membrane of the containment vehicle, to release the
cells and/or their cellular products.
[0013] Thus, an object of the present invention is to provide a
system and method for the treatment (including prevention) of
coronary artery disease, for example, by producing and delivering
locally a therapeutic agent, such as an anticoagulant.
Significantly, the local dosage can be controlled and provided on
demand without worrying about the systemic effects. Furthermore,
the local dosage can be administered prior to significant
physiological damage occurs to the patient.
[0014] In a preferred embodiment, the present invention provides an
implantable system that includes: a delivery device comprising a
carrier for carrying eukaryotic cells that produce at least one
therapeutic agent; a stimulating element for stimulating the
release of the therapeutic agent from the delivery device; and a
sensing element for monitoring at least one physiological property
of a patient in which the system is implanted and communicating
with the stimulating element to stimulate the release of the
therapeutic agent from the delivery device.
[0015] In another embodiment, an implantable system includes a
delivery device comprising an intraluminal stent, which includes a
lumen-wall contacting surface, a lumen-exposed surface, a first
polymer composition covering at least a portion of the stent
(preferably, at least a portion of the lumen-exposed and the
lumen-wall contacting surfaces), and endothelial cells associated
with the first polymer composition covering, wherein the
endothelial cells produce at least one therapeutic agent. The
implantable system further includes an electrical stimulating
element for stimulating the release of the therapeutic agent from
the delivery device, and a sensing element for monitoring at least
one physiological property of a patient in which the system is
implanted and communicating with the electrical stimulating element
to stimulate the release of the therapeutic agent from the delivery
device.
[0016] The present invention also provides a method of local
delivery (as opposed to systemic delivery) of a therapeutic agent.
The method involves simply implanting an implantable system
described above. Once implanted into a patient, the therapeutic
agent is released from the delivery device when the sensing element
detects a predetermined level of a physiological property (e.g., a
certain pH or level of blood flow or blood gases) and communicating
with the stimulating element to trigger the release.
[0017] A method of making an implantable system for local delivery
of a therapeutic agent is also included within the scope of the
present invention. The method includes: providing a delivery device
comprising a carrier and eukaryotic cells associated therewith that
produce at least one therapeutic agent; providing a stimulating
element for stimulating the release of the therapeutic agent from
the delivery device; providing a sensing element for monitoring at
least one physiological property of a patient in which the system
is implanted; and connecting the stimulating element and sensing
element such that they communicate with each other to stimulate the
release of the therapeutic agent from the delivery device when
implanted in the body of a patient and the sensing element detects
a predetermined level of a physiological property. Preferably, the
step of providing a delivery device includes: providing a carrier;
isolating eukaryotic cells from a patient; culturing the eukaryotic
cells; delivering nucleic acid of a desired therapeutic agent to
the eukaryotic cells to form genetically engineered eukaryotic
cells; and contacting the carrier with the genetically engineered
eukaryotic cells.
[0018] In an alternative embodiment, the present invention provides
implantable systems as described above without the sensing element.
In such systems, the patient in which the system is implanted
activates the stimulating element when desired (i.e., on demand).
Typically, this occurs when the patient detects a change in a
physiological condition (e.g., angina) and communicates with the
electrical stimulating element to trigger release of the
therapeutic agent. The patient can communicate with the electrical
stimulating element to activate it using radio frequency, infrared,
or acoustic pulses.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1 is an illustration of a method of making and
implanting a drug-loaded stent of the present invention.
[0020] FIG. 2 is an illustration of an implantable system according
to the present invention that includes the use of an RF signal to
communicate and generate an electrical current in a coiled stent.
Inset FIG. 2A is a diagramatic representation of a circuit in a
coiled stent for electrically stimulating the cells in association
with the stent.
[0021] FIG. 3 is an elevational view of a preferred balloon
catheter with stent including fibrin and drug-eluting cells
according to the present invention.
[0022] FIG. 4 illustrates a method of loading a stent with
drug-eluting cells.
[0023] FIG. 5 shows a circuit for an implantable medical device
according to the present invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0024] The present invention provides implantable systems for the
treatment (including prevention) of a variety of disorders, such
as, coronary artery disease, which can be manifested by stenosis,
myocardial infarction, aneurysm, angina, and/or atherosclerosis, or
cerebro-vascular occlusion, which can result in a stroke, for
example. The implantable systems of the present invention include a
delivery device comprising a carrier (e.g., stents, vascular
grafts, stent grafts) and eukaryotic cells (e.g., genetically
engineered endothelial cells), which can optionally be enclosed
within containment vehicles. Such cells are capable of producing
one or more therapeutic agents (e.g., proteins and other cellular
products) that have a preventative, therapeutic, or
disease-treating effect on surrounding tissue.
[0025] The therapeutic agents are released from the delivery device
upon the application of a stimulus (e.g., electrical stimulus).
This can occur as a result of the stimulus acting directly on the
cells to cause them to produce, activate, and/or release the
therapeutic agents. Alternatively, this can occur as a result of
the stimulus acting on the optional containment vehicles within the
delivery device, for example. Significantly, the local dosage can
be controlled and provided on demand without worrying about the
systemic effects, and before significant damage occurs to the heart
muscle, for example.
[0026] Referring to FIG. 1, in one preferred embodiment,
endothelial cells 12 are obtained from a patient 10 and grown in
cell culture 14. During proliferation in cell culture 14, cells are
infected with a genetically engineered retrovirus which integrates
the gene for the drug to be locally delivered into the chromosomes
of the endothelial cells. A cylindrical coil shaped stent 18 is
produced out of a conductive metal and coated with an insulative
material, such as an organic polymer, except at its ends, and with
genetically engineered endothelial cells 19. The stent 18 coated
with endothelial cells is introduced into the patient 10.
Typically, the coated stent is implanted into the appropriate
coronary artery during a catheterization procedure, into the right
or left coronary artery depending on the pathophysiology.
[0027] Referring to FIG. 2, a preferred embodiment of the
implantable system 20 of the present invention includes, in
addition to the coated stent 18, a stimulation device 22 that
includes, for example, an implantable generator, which can be
implanted in the left pectoralis major area, a traditional
pacemaker implant site, along with an isolated lead or antenna 24
traveling to the right ventricle via the subclavian vein, superior
vena cava, and tricuspid valve, a traditional pacemaker lead
placement technique. The stimulation device 22 can monitor the
patient (using a sensing element incorporated within the device
22), if desired, and trigger the stent (using a stimulating element
incorporated within device 22) with engineered endothelial cells to
release the cellular product(s). Typically, as is shown in this
embodiment, the stimulation device is remote from the delivery
device.
[0028] Alternatively, the patient can control the stimulation
device and cause the stent to release the therapeutic agent(s) upon
sensing a physiological change (e.g., angina) using a patient
activator to communicate with the stimulating element. In this
embodiment, the stimulation device does not need a sensing element;
rather, the patient acts as the sensor to detect an undesirable or
a change in a physiological condition. Such patient activators can
involve the use of radio frequency, infrared, acoustic pulsed, or
magnetic means. For example, upon experiencing angina (i.e., chest
pain), a patient can hold a hand-held device in place over the
implant site of the stimulating element (e.g., an implantable pulse
generator) to communicate with it and activate it to stimulate the
remote endothelial cells.
[0029] The sensing element can be in the form of electrocardiogram
(ECG) for monitoring changes in the circulatory system, such as a
reduction of blood flow in the coronary sinus, for example. A
sudden change in ECG morphology such as ST segment elevation is
usually due to the onset of ischemia caused by a decrease in flow
due to an occlusion resulting from the rupture of an unstable
plaque (i.e., a blood clot). Other sensors, such as blood gas
sensors, for example, can alternatively or additionally be used to
detect changes in a patient's coronary circulatory system. Once the
onset of ischemia is detected, an implantable pulse generator
(IPG), for example, will trigger the delivery device (preferably,
by stimulating the eridothelial cells) to release one or more
cellular products (e.g., tissue-type Plasminogen Activator or t-PA)
to dissolve the clot and prevent the infarction from taking
place.
[0030] In one preferred embodiment, as illustrated by FIG. 2, when
it is time to reduce the coagulation in a coronary artery (as
determined by a sensing element in an implantable stimulation
device 22 or by a patient experiencing angina), a radio frequency
signal 26 is sent to the coil formed by the stent 18. Because the
ends of the stent 18 are not insulated, electromotive force (EMF)
generated by the RF signal is delivered to the drug-eluting cells.
Upon that event, the cells, e.g., genetically engineered
endothelial cells, can be stimulated to release their stores of
thrombolytic agents, such as tissue-type plasminogen activator,
immediately.
[0031] Placement of genetically engineered endothelial cells, for
example, upstream in the coronary artery will allow the secretion
of various therapeutic agents (i.e., drugs) into the lumen, and the
blood will carry them through the rest of the circulatory system.
Although this eventually results in more of a systemic delivery,
the administration of the therapeutic agent is initiated as local
delivery at the site of the delivery device. Release of tissue-type
plasminogen activator (t-PA) by engineered endothelial cells in a
coronary artery would allow a potent dose of the drug to be
delivered locally while avoiding dilution, and potential toxicity
as in the case of systemic administration of the same drug.
Ideally, this system will dissolve blood clots, for example, as
soon as they form through the localized delivery of potent
quantities of t-PA, for example, upstream from the site of thrombus
formation.
[0032] Drug-Eluting Cells
[0033] Cells suitable for use in the present invention include a
wide variety of eukaryotic cells that produce therapeutic agents,
or can be genetically engineered to produce therapeutic agents.
Ideally, such cells are also able to secrete these agents,
particularly upon the application of a stimulus, such as an
electrical stimulus. Suitable cells for use in the present
invention typically include mesenchymal or mesodermal cells,
including, but not limited to endothelial cells and fibroblasts,
whether they are autologous or allogeneic, genetically engineered
or nonengineered. Mixtures of such cells can also be used.
Endothelial cells are particularly suitable for use in the present
invention because they line the walls of the blood vessels. They
are also particularly suitable for use in the present invention
because they are capable of secreting vasodilatory, thrombolytic,
or angiogenic factors, which can facilitate recovery of ischemic
myocardium.
[0034] Endothelial cells and fibroblasts are preferred because they
have been shown to be suitable for use in ex vivo gene transfer. Ex
vivo gene transfer (also referred to herein as ex vivo gene
therapy) is a process by which cells are removed from the body
using well known techniques, genetically manipulated, usually
through transduction or transfection of nucleic acid into the cells
in vitro, and then returned to the body for therapeutic purposes.
This contrasts with in vivo gene therapy, where a gene transfer
vector is administered to the patient resulting in genetic transfer
into cells and tissues in the intact patient. Ex vivo techniques
are well known to one of skill in the art.
[0035] Ex vivo gene therapy is an effective approach because the
target cells to be used in the procedure can be manipulated as
needed to optimize gene transfer efficiency and thus the
effectiveness of the overall procedure. However, the ex vivo
approach can only be utilized for those cell types which can be
readily retrieved from the body, cultured ex vivo, and then
returned to the body. Such cells include blood and marrow cells,
liver hepatocytes, skin fibroblasts, muscle myoblasts, and vascular
endothelial cells. Thus, endothelial cells and fibroblasts, which
can be efficiently infected by retroviral vectors in vitro, and
then transplanted back into the host to achieve gene transfer in
vivo, are particularly preferred for use in the present
invention.
[0036] Autologous endothelial cells are particularly desirable.
Vascular endothelial cells have been removed from a patient and
transduced ex vivo with a retroviral vector designed for expression
of .beta.-galactosidase as a reporter gene, as disclosed in Nabel
et al., Science, 244, 1342-1344 (1989). Such genetically engineered
cells can be reintroduced into a patient. For example, Nabel et al.
showed that endothelial cells engineered to express
.beta.-galactosidase could be introduced into an occluded vascular
site using a balloon catheter, after which the presence of
engrafted endothelial cells could be detected by staining for
.beta.-galactosidase activity.
[0037] In one embodiment of the present invention, endothelial
cells are obtained from a patient and grown in cell culture. During
proliferation in cell culture, they are infected with a genetically
engineered retrovirus which integrates the gene for the drug to be
locally delivered into the chromosomes of the endothelial cells.
This can be accomplished, for example, according to the teachings
of U.S. Pat. No. 5,674,722 (Mulligan et al.) and Dichek et al.,
Mol. Biol. Med., 8, 257-266 (1991). For the treatment of coronary
artery disease (CAD), candidate genes include the gene encoding
wild-type tissue plasminogen activator and the gene encoding
protein C, for example. The isolation and characterization of the
human t-PA structural gene is disclosed in Fisher et al., J. Biol.
Chem., 260, 11223-11230 (1985).
[0038] Activated Protein C degrades the coagulation factors which
are responsible for the formation of life threatening clots in the
coronary arteries. However, protein C requires the presence of
Ca.sup.++ (calcium ion) for its own activation, which is controlled
in the cells by ion gates which, in turn, are under the control of
the voltages across the cell membrane. Thus, electrical stimulation
of the cells can activate protein C. For example, endothelial cells
may be depolarized, thereby altering the ion concentrations within
the cells. Although the inventors do not wish to be bound by
theory, it is believed that changing ion concentration, such as
calcium ion concentration, may trigger secretion of certain
cellular products, such as t-PA. Alternatively, calcium ions can
activate protein C, which can start the cascade for the reduction
of coagulation factors and prevent the formation of clots.
[0039] There are a wide variety of methods that can be used to
deliver nucleic acid to the eukaryotic cells if they are to be
modified to secrete a desired agent. These are well known to one of
skill in the art. The desired nucleic acid can be inserted into an
appropriate delivery vehicle, such as, for example, an expression
plasmid, cosmid, YAC vector, and the like. There are a number of
viruses, live or inactive, including recombinant viruses, that can
also be used. A retrovirus can be genetically modified to deliver
any of a variety of genes. Adenovirus has been used in a variety of
experiments to deliver nucleic acid capable of directing and
expressing protein in a cell. These include, but are not limited
to, superoxide dismutase, tissue plasminogen activator, and
interleukin-10.
[0040] Exemplary nucleic acid that would function as nucleic acid
for incorporation into the cells include, but are not limited to,
nucleic acid operably encoding a protein, polypeptide, or peptide
to deliver a therapeutic effect to a cell. The nucleic acid can
include an entire gene or a portion of a gene. Exemplary genes
include, but are not limited to, the active form of the nitric
oxide synthase (a protein that is known to relax blood vessels and
prevent clot formation), and prostaglandin H synthase (to restore
an endogenous inhibitor of platelet aggregation and
vasoconstriction following injury to endothelium).
[0041] There are a variety of disorders that can be treated using
the systems and devices of this invention. Examples of these
disorders include, but are not limited to, damage associated with
myocardial infarction or aneurysms (targeting fibroblast growth
factor or transforming growth factor .beta. and protease,
respectively), atherosclerosis (targeting high density
lipoprotein), and hypercoagulable states (targeting
tissue-plasminogen activator).
[0042] The gene sequence of the nucleic acid delivered by the
virus, including nucleic acid encoding proteins, polypeptide or
peptide is available from a variety of sources including GenBank
(Los Alamos National Laboratories, Los Alamos, N. Mex.), EMBL
databases (Heidelberg, Germany), and the University of Wisconsin
Biotechnology Center, (Madison, Wis.), published journals, patents
and patent publications. All of these sources are resources readily
accessible to those of ordinary skill in the art. The gene sequence
can be obtained from cells containing the nucleic acid fragment
(generally, DNA) when a gene sequence is known. The nucleic acid
can be obtained either by restriction endonuclease digestion and
isolation of a gene fragment, or by polymerase chain reaction (PCR)
using oligonucleotides as primers either to amplify cDNA copies of
mRNA from cells expressing the gene of interest or to amplify cDNA
copies of a gene from gene expression libraries that are
commercially available. Oligonucleotides or shorter DNA fragments
can be prepared by known nucleic acid synthesis techniques and from
commercial suppliers of custom oligonucleotides such as Amitof
Biotech Inc. (Boston, Mass.), or the like. Those skilled in the art
will recognize that there are a variety of commercial kits
available to obtain cDNA from mRNA (including, but not limited to
Stratagene, La Jolla, Calif. and Invitrogen, San Diego, Calif.).
Similarly, there are a variety of commercial gene expression
libraries available to those skilled in the art including libraries
available form Stratagene, and the like. General methods for
cloning, polymerase chain reaction and vector assembly are
available from Sambrook et al. eds. (Molecular Cloning: A
Laboratory Manual, 1989 Cold Spring Harbor Laboratory Press, Cold
Spring Harbor, N.Y.) and Innis, et al. eds. (PCR Strategies, 1995,
Academic Press, New York, N.Y.).
[0043] Depending on the maximum genome size that a particular viral
genome can accommodate or that can be associated with a virus
particle, the virus delivering nucleic acid to the cell can include
nucleic acid encoding one or more proteins, polypeptides, or
peptides. Oligonucleotides can be delivered by virus through the
incorporation of oligonucleotides within the virus or associated
with the outer surface of the virus using methods well known to one
of skill in the art.
[0044] Therapeutic agents (i.e., drugs) that can be produced,
stored, and secreted by eukaryotic cells, particularly genetically
engineered endothelial cells, include, but are not limited to,
anticoagulant agents, antiplatelet agents, antifibrinolytic agents,
angiogenesis factors, etc. Specific examples include activated
protein C, tissue plasminogen activator, prostacyclin, and vascular
endothelial growth factor (VEGF). Such cells are referred to herein
as "drug-eluting" cells.
[0045] The drug-eluting cells can be coated directly on the carrier
of the delivery device or they can be incorporated into the
carrier, as in a polymer film (i.e., sheeting) or coating on a
stent. They can also be included within a microscopic containment
vehicle that can be stimulated to release the cells and/or their
cellular products. The microscopic containment vehicle can be
coated on the carrier or incorporated into the carrier, as in a
polymer film or coating on a stent. The cells can be induced to
produce, activate, and/or release their cellular products
(including one or more therapeutic agents) by an external
stimulation device. This can occur as a result of destruction of
the cellular membrane, for example. Alternatively, cells can
constitutively release low levels of one or more therapeutic agents
all the time. These cells would be preferentially placed in a
containment vehicle, which would then be induced to release the
therapeutic agent by an external stimulation device.
[0046] Delivery Devices
[0047] The drug-eluting cells can be carried by a variety of
carriers for use in the delivery devices of the present invention.
For example, intravascular stents and vascular grafts have been
seeded with endothelial cells expressing recombinant t-PA, as
disclosed in Dichek et al., Circulation, 8, 1347-1353 (1989) and
Dichek et al., Circulation, 93, 301-309 (1996). In addition to
stents and vascular grafts or stent grafts, suitable carriers for
use in delivery devices of the present invention include, but are
not limited to, synthetic patches, infusion sleeves, medical
electrical leads and electrodes, and indwelling catheters.
[0048] Although any of these devices can be used as carriers for
the drug-eluting cells, the following description focuses on
stents. Whether the carrier is a stent or other medical device, the
drug-eluting cells can be directly coated onto the carrier,
incorporated into or onto a polymeric coating or film on the
carrier, incorporated into microscopic containment vehicles and
coated directly on the carrier or incorporated into or onto a
polymeric coating or film on the carrier. As used herein, a
"delivery device" includes the carrier (e.g., stent), cells that
produce one or more therapeutic agents, and optional containment
vehicles, as well as other optional therapeutic materials.
[0049] The term "stent" refers to any device capable of being
placed into contact with a portion of a wall of a lumen. It
includes classical stents used in intravascular applications, as
well as any prosthesis that may be inserted and held where desired
in a lumen. Such prostheses include single- or multi-filar braided
mesh designs, metallic malleable designs, and others, as disclosed,
for example, in International Patent Application No. WO 91/12779
(Medtronic, Inc.).
[0050] Typically, the stent has a lumen wall-contacting surface and
a lumen-exposed surface. Where the stent is shaped generally as a
tube-like structure, including a discontinuous tube or a ring-like
structure, the lumen-wall contacting surface is the outside surface
of the tube and the lumen-exposed surface is the inner surface of
the tube. The stent can include polymeric elements, metallic
elements, filametary elements, or combinations thereof. It can be
coated with fibronectin or other extracellular matrix for adhering
the cells to the stent. Preferably, the stent includes a fibrin or
collagen coating or film or other natural or synthetic polymeric
coating or film for holding the cells in place.
[0051] A deformable metal wire stent useful as a stent framework to
support the cells of this invention is disclosed in U.S. Pat. No.
4,886,062 (Wiktor). Other metallic stents useful in this invention
include those of U.S. Pat. No. 4,733,655, (Palmaz) and U.S. Pat.
No. 4,800,882 (Gianturco). FIG. 3 provides an elevational view of a
preferred stent 30 of this invention. In one preferred embodiment,
the stent comprises a stent framework 32 with a polymer film 34
(preferably, a fibrin film) extending circumferentially around at
least a portion of the lumen-wall contacting surface of the stent
and/or the lumen-exposed surface of the stent and preferably
extending over substantially all of the lumen-wall contacting and
lumen-exposed surfaces. A balloon 36 is positioned in FIG. 3
adjacent the lumen-exposed surface of the stent to facilitate
delivery of the stent. Polymeric stents can also be used in this
invention, whether they be nonbioabsorbable or bioabsorbable in
part, or total.
[0052] The drug-eluting cells can be coated directly on the stent
or they can be incorporated into a polymer film (i.e., sheeting) or
coating. The stents of this invention preferably include a first
polymer composition comprising fibrin or other polymer to provide
containment for the drug-eluting cells. The first polymer
composition of this invention can be prepared from a homopolymer, a
copolymer (i.e., a polymer of two or more different monomers), or a
composition (e.g., a blend) comprising fibrin with one or more
polymers or copolymers, for example. The composition preferably
forms a viscoelastic, tear-resistant, biocompatible polymer. The
term "viscoelastic" refers to the prescribed "memory"
characteristics of a molecule that allow the molecule to respond to
stress as if the molecule was a combination of elastic solids and
viscous fluids. The term "tear resistant" indicates that when the
polymer is exposed to expansion stress, the material does not
substantially tear. Tearing refers to the propagation of a nick or
cut in the material while under stress. When the stent of this
invention is expanded on a balloon, the polymer is able to expand
to accommodate the balloon expansion. The term "biocompatible" is
used herein to refer to a material that does not have toxic or
injurious effects on biological systems.
[0053] Preferably, the first polymer composition minimizes or does
not exacerbate irritation to the lumen wall when the stent is in
position. The first polymer composition is preferably
nonthrombogenic when applied alone or alternatively when used with
anti-thrombogenic agents such as heparin, and the like, or with
anti-inflammatory agents such as cyclosporins, and the like. The
first polymer composition is also preferably a biostable or a
bioabsorbable polymer depending on the desired rate of release or
the desired degree of polymer stability.
[0054] The stents of this invention can be coated with fibrin, for
example, by application of a fibrinogen solution and a solution of
a fibrinogen-coagulating protein or by attachment of a fibrin
preform or sleeve such as an encircling film of fibrin, including
the film provided in U.S. Pat. No. 4,548,736 (Muller). As described
in U.S. Pat. No. 4,548,736 (Muller) and U.S. Pat. No. 5,510,077
(Dinh et al.), fibrin is clotted by contacting fibrinogen with a
fibrinogen-coagulating protein such as thrombin. The fibrinogen is
preferably used in solution with a concentration of about 10 to
about 50 mg/ml with a pH of about 5.89 to about 9.0 and with an
ionic strength of about 0.05 to about 0.45. The fibrinogen solution
typically contains proteins and enzymes such as albumin,
fibronectin, Factor XIII, plasminogen, antiplasmin, and
Antithrombin III. The thrombin solution added to make the fibrin is
typically at a concentration of up to about 120 NIH units/ml with a
preferred concentration of calcium ions between about 0.02 M and
0.2 M. Also preferably, the fibrinogen and thrombin used to make
fibrin in the present invention are from the same animal or human
species as that in which the stent of the present invention will be
implanted to avoid cross-species immune reactions. The resulting
fibrin can also be subjected to heat treatment at about
1500.degree. C., for about 2 hours to reduce or eliminate
antigenicity.
[0055] The fibrin, or other polymer, can be in the form of a film
produced by casting the combined fibrinogen and thrombin in a film
and removing moisture from the film osmotically through a moisture
permeable membrane. Alternatively, a substrate can be contacted
with a fibrinogen solution and with a thrombin solution resulting
in a fibrin film formed by polymerization of fibrinogen on the
surface of the device. Multiple layers of fibrin applied by this
method can provide a fibrin film in a variety of thicknesses. In
another method, the fibrin can be first clotted and then ground
into a powder that is mixed with water and stamped into a desired
shape in a heated mold. These methods can be used with fibrin
monomers or with combinations of monomers to form the first polymer
composition of this invention. Those skilled in the art will
recognize that the methods for forming the first polymer
composition can be modified to include other polymers, as
contemplated in this invention, without undue experimentation.
[0056] The first polymer compositions of this invention can include
one or more other synthetic or natural polymers. Suitable polymers
include those that are compatible with the cells and therapeutic
agents. These include, but are not limited to, fibrins, collagens,
alginates, polylactic acids, polyglycolic acids, celluloses,
hyaluronic acids, polyurethanes, silicones, polycarbonates,
mixtures or copolymers thereof, and a wide variety of other
polymers typically disclosed as being useful in implantable medical
devices. An example of a copolymer with improved structural
strength and improved biological performance is a fibrin and
polyurethane copolymer, as disclosed in U.S. Pat. No. 5,510,077
(Dinh et al.), or a fibrin alginate copolymer. Because fibrin is
more readily degraded in the body than polyurethane, polyurethane
can be used to regulate degradation of the fibrin covering the
stent and to slow release of the cellular products from the
stent.
[0057] Heparin, or other anticoagulants, such as polyethylene
oxide, hirudin, and tissue plasminogen activator, can be
incorporated into the stent prior to implantation in an amount
effective to prevent or limit thrombosis. A heparin immersion
procedure can be used to incorporate the heparin. A heparin
solution can alternatively be added at the time that cells are
loaded onto the stent. Heparin can also be incorporated into the
polymer matrix before it has completely polymerized. For example,
powdered heparin can be dusted onto the stent during the
polymerization process and additional thrombin and fibrinogen can
then be applied as a coating over the heparin.
[0058] The shape of the polymer film or coating can be modified
based on the methods used to cover the stent. It can be spray
coated onto the stent or a film can be molded over the stent
framework. The first polymer composition of this invention can
cover both the lumen wall contacting surface and the lumen-exposed
surface of the stent.
[0059] Alternatively, a porous polymeric sheet material can be used
into which fibrin is incorporated. The sheet could be prepared from
polyurethane, for example, by dissolving a polyether urethane in an
organic solvent such as 1-methyl-2-pyrrolidone; mixing into the
resulting polyurethane solution a crystalline particulate material
like salt or sugar that is not soluble in the solvent; casting the
solution with particulate material into a thin film; and then
applying a second solvent, such as water, to dissolve and remove
the particulate material, thereby leaving a porous sheet. The
porous sheet could then be placed into a fibrinogen solution in
order to fill the pores with fibrinogen followed by application of
a solution of thrombin and fibrinogen to the surface of the sheet
to establish a fibrin matrix that occupies both the surface of the
sheet and the pores of the sheet. Preferably, a vacuum would be
pulled on the sheet to insure that the fibrinogen applied to the
sheet is received into the pores.
[0060] The stent framework can also be positioned within a mold and
the compounds forming the first polymer composition incorporated
into the mold. The first polymer composition forming a sheet or
sleeve can be prepared in an extended shape and then compressed or
dehydrated into a final shape to fit over the stent. In this way,
when the stent is expanded in place to fit the walls of a lumen,
the first polymer composition can be readily expanded without
tearing or introducing irregularities into the sleeve and/or the
coating.
[0061] The stent can be loaded with drug-eluting cells by mixing
the monomer solution of the first polymer composition with cells or
by directly applying the cells to the polymerized composition. In a
first embodiment, the stent is loaded with cells at the time of
formation of the first polymer composition. FIG. 4 provides an
example where a stent is formed over a balloon and introduced into
a mold cavity to receive a solution sufficient to form the first
polymer composition and including cells to be incorporated into the
first polymer coating. In FIG. 4, the stent framework 42 is
positioned over the balloon 46 and introduced into a mold 48. A
monomer solution capable of forming a first polymer composition is
introduced into the mold along with the drug-eluting cells. Once
the polymer is formed over the stent framework 42, the stent is
released from the mold. The drug-eluting cells can also be included
in a monomer solution as a spray or liquid coating to be applied to
the stent framework.
[0062] Alternatively, the drug-eluting cells can be added to the
polymer coated stent either at the time of stent manufacture or by
the physician, prior to stent implantation. Where the first polymer
composition is capable of dehydration and rehydration, the fibrin
coated stent can be supplied in a sterile, dehydrated form and
cells can be loaded onto the stent by rehydration of the first
polymer composition positioned on the stent framework by immersing,
wicking, or spraying a liquid suspension (such as a balanced salt
solution, including HBSS or a tissue culture media, and the like)
containing the cells onto the stent prior to stent implantation.
The liquid used to deliver cells to the first polymer coating
should support the stability of the cells and should include pH
buffered solutions, stabilizers such as albumin, glycerol, or the
like, in a form that is biocompatible and preferably
nonimmunogenic.
[0063] In one embodiment, a dehydrated first polymer covered stent
can be rehydrated in a solution of calcium chloride and sodium
alginate containing the drug-eluting cells. The solution can be
sprayed onto the stent, for example. A layer of calcium alginate
then precipitates onto the surface of the stent with the cells.
Alternatively, a dehydrated first polymer covered stent can be
rehydrated in a solution of sodium alginate containing the
drug-eluting cells, and then a layer of calcium alginate is sprayed
or otherwise coated over the first polymer covering and cells.
Where the cells are incorporated into a polymer solution for spray
application or air pump applications, the polymer should be of
sufficiently low viscosity to facilitate the application of the
solution to the stent.
[0064] In another embodiment of the stents of this invention, a
second polymer composition can be added over the first polymer
composition following, or at the time of, cell loading. Preferably,
the second polymer composition is biodegradable and the second
polymer composition coats at least a portion of the first polymer
composition and cells. For example, after the cells are loaded onto
a stent having a first polymer composition coated thereon, the
stent can be sprayed with or immersed in a solution to form a
coating of a biodegradable polymer such as polylactic acid or
methylcellulose, with or without heparin, or another coagulation
inhibitory or anti-inflammatory compound. Advantageously, the
second covering of polymer provides even greater sustained release
capabilities and prevents loss of cells from the coated or covered
stent during transit through the body to the target site on the
body lumen. Suitable polymers for use in the second polymer
composition include, but are not limited to, those listed above for
the first polymer composition, and combinations thereof, and a wide
variety of others typically disclosed as being useful in
implantable medical devices.
[0065] One or more surfaces of the stent can be coated with one or
more additional coatings of a polymer that is the same or different
from the second polymer composition. Additional polymer coatings on
the lumen-exposed surface are used to prevent release of the cells
from the lumen-exposed surface of the stent when the stent is
positioned in the body. This embodiment is particularly useful
where the stent is used in the blood vasculature and in particular
where the stent is used in the coronary artery.
[0066] The drug-eluting cells can also be included within a
microscopic containment vehicle that can be stimulated to release
the cells and/or their cellular products. This microscopic
containment vehicle can be coated onto the delivery device directly
or into or onto a polymer coating or film. The drug-eluting cells
are enclosed within such a vehicle. Upon stimulation of the
containment vehicle, the cells and/or cellular products (e.g.,
therapeutic agents) are released from the containment vehicle. Such
containment vehicles are particularly desirable for cells that
continuously produce and release the therapeutic agent. Thus,
instead of stimulating the cells themselves, the containment
vehicle is stimulated to release the cells and/or therapeutic
agents. Stimulation of the containment vehicles can be accomplished
using a variety of techniques.
[0067] Examples of microscopic containment vehicles include, but
are not limited to, those described in International Patent
Application Nos. WO 96/28841 (Ohman) and WO 96/34417 (Smela et
al.), and in Smela et al., Science, 268, 1735-1738 (1995), which
disclose micromachined structures and microactuators. In one
example, such micromachined structures include conducting bilayers
made of a layer of polymer and a layer of gold that are used as
hinges to connect rigid plates to each other and to a silicon
substrate. The bending of the hinges is electrically controlled and
reversible, allowing precise three-dimensional positioning of the
plates. The use of differential adhesion allows the structures to
be released from the silicon substrate. Thus, complex shapes can be
formed, such as cubes, of micrometer size that could be used to
contain drug-eluting cells and their cellular products in
association with a delivery device such as a stent, until they are
needed for therapeutic or preventative treatment.
[0068] Microscopic containment vehicles can also include
micropumps, reservoirs, with piezoelectric valves. The body of the
pump and reservoir can be made of a polymeric material while the
valves contain a piezoelectric material, which allows opening and
closing of the valves and pumping movement by electrical
stimulation. The reservoirs contain the drug-eluting cells and
their cellular products. Upon stimulation of the piezoelectric
material, the valve is opened to release the cells arid/or cellular
products. Implantable piezoelectric pumps are known (see, for
example, U.S. Pat. No. 4,944,659 (Labbe et al.)) and can be
modified by one of skill in the art to form containment vehicles
for the present invention.
[0069] The stents of this invention can be provided in a sterile,
dehydrated form, in a hydrated form with cells (shipped frozen or
on ice) or as a first polymer covered stent supplied with the
necessary materials to facilitate cells loading and further coating
or covering of the stent as needed. Therefore, this invention also
relates to a kit comprising a stent with a first polymer
composition comprising fibrin, buffers suitable for rehydrating the
stent and loading the cells and a container to facilitate sterile
loading of the stent. Optionally, the kit can include further
coatings or coverings to be applied over the first polymer
composition. In a preferred embodiment, the kit includes: a stent
comprising a lumen-wall contacting surface, a lumen-exposed
surface, and a first polymer composition comprising fibrin covering
at least a portion of the stent; a cell-loading composition to be
applied to the stent; and a container to house the stent and the
composition during application of the cell-loading composition.
[0070] Stimulating and Sensing Elements
[0071] Systems of the present invention include a second
implantable device (i.e., a stimulation device) that includes a
stimulating element, such as an implantable pulse generator (IPG)
similar in many respects to pacemakers and defibrillators known in
the art, for example, which is preferably in contact with a sensing
element. This device monitors the patient to detect when a stimulus
needs to be sent to the cells to trigger release of one or more
therapeutic agents. This monitoring can be in the form of an
electrocardiogram (ECG), for example, to detect an ST segment
elevation or a reduction of blood flow in the coronary sinus. Once
the onset of ischemia is detected, the stimulating element will
trigger the release of cellular products. For example, when a blood
clot is formed that reduces blood flow, an abnormal ECG is
produced, which causes an IPG, for example, to trigger a stent to
stimulate endothelial cells to release t-PA, which travels to the
blood clot and dissolves it, thereby preventing myocardial
infarction.
[0072] Cellular products can be released upon the action of a
stimulus directly on the cells, particularly the cellular membrane,
or on the microscopic containment vehicle in which the cells are
optionally located. The cellular products, particularly the
therapeutic agents, are then distributed to the desired site
through a washing action of the delivery device with body fluids.
Following stimulation of the cells or containment vehicle, cellular
products can also be released by erosion or bioabsorption of the
coated surfaces of the stent, for example, by dissolution of fibrin
or hydrolysis resulting in crosslink cleavage and concomitant
release of the physically entrapped cells.
[0073] The stimulation device can provide electrical stimulation,
mechanical stimulation, acoustic stimulation, thermal stimulation,
chemical stimulation, or combinations thereof, to the eukaryotic
cells and/or the containment vehicles, for example. One of skill in
the art will recognize that a variety of means of stimulation can
be used to electrically, mechanically, acoustically, and/or
chemically stimulate the cells and/or the containment vehicles. The
following are presented as examples only and do not limit the
specific mechanisms or devices by which the stimulation can
occur.
[0074] Typically, electrical stimulation causes ions to flow
through cells making their membranes permeable. If sufficient
stimulation is applied, then large pores are created on the
cellular membrane, which are sufficient for the cellular product to
pass through. Electrical stimulation can occur as described herein
with reference to FIG. 2.
[0075] Mechanical stimulation can be applied by twisting the
cellular membranes of the cells, for example. This causes a shear
force on the cells and elongates them. Surface area is not changed
but the enclosed volume is decreased so the intracellular pressure
increases, causing the contents of the cells to leak out.
Alternatively, mechanical stimulation can be accomplished through
the use of a piezoelectric material (e.g., a piezoelectric crystal)
on the surface of the carrier of the to delivery device. When an
electrical pulse is applied to the crystal, it changes size (e.g.,
longer or shorter), which can stretch or compress the carrier, and
hence cause pressure on the cells. Acoustic stimulation is similar
to mechanical stimulation, but the pressure is applied repetitively
by sound waves.
[0076] Thermal stimulation involves applying heat to the cells,
which can cause them to release their cellular products. This can
be accomplished by the use of resistive elements inside the carrier
of the delivery device. When electrical current is applied, the
resistive element applies heat to the cells.
[0077] Chemical stimulation involves releasing a compound (e.g., a
ligand) into the blood stream that acts via a lock and key
mechanism to interact with the cells. For example, a ligand can be
released from the stimulation device and when it reaches the cells
in the delivery device, it can attach to an appropriate receptor on
the cells and trigger events leading to the release of the
therapeutic agent.
[0078] Although current implantable pulse generators (IPGs) are
designed to stimulate cardiac muscle tissue, they may be modified
readily by one of skill in the art to stimulate drug-eluting cells
in accordance with the teachings of the present invention. It will
be appreciated that the stimulation device according to the present
invention can include a wide variety of microprocessor-based
implantable stimulators similar to those used in pacemakers, as
disclosed in U.S. Pat. No. 5,158,078 (Bennett et al.), U.S. Pat.
No. 5,312,453 (Shelton et al.), and U.S. Pat. No. 5,144,949
(Olson), and pacemaker/cardioverter/defibrillators (PCDs), as
disclosed in U.S. Pat. No. 5,545,186 (Olson et al.), U.S. Pat. No.
5,354,316 (Keimel), U.S. Pat. No. 5,314,430 (Bardy), U.S. Pat. No.
5,131,388 (Pless), and U.S. Pat. No. 4,821,723 (Baker et al.).
Alternatively, the stimulation device can include stimulating
elements similar to those used in implantable nerve or muscle
stimulators, such as those disclosed in U.S. Pat. No. 5,199,428
(Obel et al.), U.S. Pat. No. 5,207,218 (Carpentier et al.), and
U.S. Pat. No. 5,330,507 (Schwartz).
[0079] In general, the implantable stimulation device 22 shown in
FIG. 2 includes a hermetically sealed enclosure that may include
various elements, such as an electrochemical cell (e.g., a lithium
battery) for providing electrical current to circuitry, circuitry
that controls device operations, a telemetry transceiver antenna,
and a circuit that receives downlinked telemetry commands from and
transmits stored data in a telemetry uplink to an external
programmer, in addition to other elements.
[0080] FIG. 5 is a block diagram illustrating various components of
an implantable stimulation device 22 which is programmable by means
of an external programming unit (not shown). One such programmer
easily adaptable for the purposes of the present invention is the
commercially available Medtronic Model 9790 programmer. The
programmer is a microprocessor device which provides a series of
encoded signals to stimulation device 22 by means of a programming
head which transmits radio frequency encoded signals according to a
telemetry system, such as that described in U.S. Pat. No. 5,312,453
(Wyborny et al.), for example.
[0081] Stimulation device 22, illustratively shown in FIG. 5 as an
exemplary embodiment, is electrically coupled to lead or antenna
24. Lead 24 may be used for stimulating only, or it may be used for
both stimulating and sensing. Lead 24 is coupled to a node 62 in
the circuitry of stimulation device 22 through input capacitor 60.
Input/output circuit 68 also contains circuits for interfacing with
stimulation device 22, antenna 66, and circuit 74 for application
of stimulating signals to lead 24 under control of
software-implemented algorithms in microcomputer unit 78.
[0082] Microcomputer unit 78 comprises on-board circuit 80 which
includes system clock 82, microprocessor 83, and on-board RAM 84
and ROM 86. In this illustrative embodiment, off-board circuit 88
comprises a RAM/ROM unit. On-board circuit 80 and off-board circuit
88 are each coupled by a data communication bus 90 to digital
controller/timer circuit 92. The electrical components shown in
FIG. 5 are powered by an appropriate implantable battery power
source 94 in accordance with common practice in the art. For
purposes of clarity, the coupling of battery power to the various
components of stimulating element 22 is not shown in the
figures.
[0083] Antenna 66 is connected to input/output circuit 68 to permit
uplink/downlink telemetry through RF transmitter and receiver unit
55. Unit 55 may correspond to the telemetry and program logic
disclosed in U.S. Pat. No. 4,556,063 (Thompson et al.), or to that
disclosed in the above-referenced Wyborny et al. patent. Voltage
reference (V.sub.REF) and bias circuit 61 generates a stable
voltage reference and bias current for the analog circuits of
input/output circuit 68. Analog-to-digital converter (ADC) and
multiplexer unit 58 digitizes analog signals and voltages to
provide "real-time" telemetry signals and battery end-of-life (EOL)
replacement functions.
[0084] Sense amplifier 53 amplifies sensed signals and provides an
amplified signal to peak sense and threshold measurement circuitry
57. Circuitry 57, in turn, provides an indication of peak sensed
voltages and measured sense amplifier threshold voltages on path 64
to digital controller/timer circuit 92. An amplified sense
amplifier signal is then provided to comparator/threshold detector
59. Sense amplifier 53 may correspond in some respects to that
disclosed in U.S. Pat. No. 4,379,459 (Stein).
[0085] Circuit 92 is further preferably coupled to electrogram
(EGM) amplifier 76 for receiving amplified and processed signals
sensed by an electrode disposed on lead 24. The electrogram signal
provided by EGM amplifier 76 is employed when the implanted device
is being interrogated by an external programmer (not shown) to
transmit by uplink telemetry a representation of an analog
electrogram of the patient's electrical heart activity. Such
functionality is, for example, shown in previously referenced U.S.
Pat. No. 4,556,063. Note that lead or antenna 24 may be located in
positions other than inside the heart.
[0086] Output pulse generator 74 provides stimuli to lead 24
through coupling capacitor 65 in response to a stimulating trigger
signal provided by digital controller/timer circuit 92. Output
amplifier 74, for example, may correspond generally to the output
amplifier disclosed in U.S. Pat. No. 4,476,868 (Thompson).
[0087] It is to be understood that FIG. 5 is an illustration of an
exemplary type of stimulation device which may find application in
the present invention, or which may be modified for use in the
present invention by one of skill in the art, and is not intended
to limit the scope of the present invention.
[0088] In addition to a stimulating element within stimulation
device 22, systems of the present invention include a sensing
element for monitoring at least one physiological property to
detect a change in a physiological condition (typically, the onset
of ischemia caused by a decrease in blood flow due to an occlusion
resulting from the rupture of unstable plaque). For example, a
pseudo-surface electrocardiogram (ECG) using a subcutaneous
electrode array can be used to detect a reduction in blood flow,
which is represented by an abnormal morphology (e.g., inverted
shape) of a T wave (i.e., the portion of an ECG pattern due to
ventricular repolarization or recovery). Such a pseudo-surface ECG
is similar to a normal ECG modified for implantation. In this
sensing element, for example, an implantable pulse generator having
three electrodes, each only about one centimeter apart, could be
implanted into the pectoralis muscle in the chest of a patient. An
ECG pattern, similar to that of a normal ECG, would be monitored
for an indication of an abnormal morphology of a T wave.
[0089] Sensing elements can include one or more individual sensors
for monitoring one or more physiological properties. In addition to
a pseudo-surface ECG, such sensors include, for example, blood gas
(e.g., CO.sub.2) sensors, pH sensors, blood flow sensors in the
coronary sinus, and the like. Other mechanisms of detection that
can be used in sensors include, for example, acoustic time of
flight changes as a result of flow, acoustic doppler, which takes
advantage of the doppler effect (received frequency is different
that the transmitted one), thermal dilution (a clinical technique
to measure blood flow and cardiac output), and venous pressure drop
due to lack of driving pressure from the blocked artery. Examples
of sensors or implantable monitoring devices that can be modified
to be used in the stimulation devices of the present invention are
disclosed, for example, in U.S. Pat. No. 5,409,009 (Olson), U.S.
Pat. No. 5,702,427 (Ecker et al.), and U.S. Pat. No. 5,331,966
(Bennet et al.). Suitable sensors and sensing techniques are well
known to one of skill in the art and can be readily adapted for use
in the present invention.
[0090] For electrical stimulation, because the direct wiring from
the stimulation device to the delivery device is not feasible, a
telemetric communication system can be used. Such systems are known
and can be modified to be used in the present invention. Examples
of such systems are discussed above and described in U.S. Pat. No.
5,312,453 (Wyborny et al.), for example. Referring again to FIG. 2,
the stimulation device 22 will produce a radio frequency (RF)
signal 26 using lead 24 as a transmitting antenna, which is
implanted into the right ventricle of a patient's heart. The lead
24 includes a coil, which transmits an H-field coupled pulse series
(e.g., a 175 kilocycle, pulse position modulated, H-field signal)
to the coil formed by stent 18, which is implanted into the left
ventricle of a patient's heart. For a conversion of the magnetic
field (H-field) into an electrical signal, the stent includes a
metal coil with all but the ends coated with an insulating coating.
A tuned LC filter integral with the stent 18 will receive some of
this signal, rectify it, and when a sufficient charge is
transferred from the stimulation device to the stent, energy will
be used to electrically stimulate the engineered endothelial cells
in the coronary artery to trigger the release of the therapeutic
agent. Inset FIG. 2A is a representation of a circuit within the
stent. This circuit includes an LC filter tuned to the specific
frequency of the RF signal 26 for induction of electric current in
the coiled stent 18 (or in a coil provided in the stent). A charge
is stored in capacitor (C.sub.storage) until a threshold voltage is
reached, which activates a switch to close the circuit and cause
current to flow through the cells associated with the stent. In
such a system, the current density produced is preferably
sufficient to stimulate the cells to release a therapeutic agent
without significant damage to the cells or the therapeutic agents.
Based on in vitro experiments, the current density for preferred
embodiments is maintained below about 540 .mu.-Amp/mm.sup.2 and the
current threshold below about 0.9 mA/mm.sup.2 to avoid damage to
the endothelial cells or the expressed product.
[0091] Thus, the delivery device that acts as a carrier for the
drug-eluting cells can be powered with RF coupling. Alternatively,
it can be self powered or battery powered.
EXAMPLES
[0092] The following examples are intended for illustration
purposes only.
[0093] Electrical Stimulation Studies
[0094] Electrical and mechanical hardware were modified to enable
the stimulation of endothelial cells cultured on inserts placed in
tissue culture wells. The cells were stimulated with single pulses
(1 milli-second long and delivered at a rate of 1 Hz) for 5 minutes
a day, on 3 consecutive days. At the end of the 3 days, cells were
removed from the culture wells, stained with crystal violet and
photographed. Increases in the current density caused a decrease in
the density of red stained endothelial cells. While a stimulation
pattern that produced 540 .mu.-Amp/mm.sup.2 of current density
appears to be detrimental to the endothelial cells, another pattern
which produced 270 .mu.-Amp/mm.sup.2 of current density did
not.
[0095] Studies continued to determine the stimulation parameters,
and upper limits on the current density to prevent cell damage. By
modifying the interface circuit between the stimulator and the
tissue culture wells, the current densities in wells where
endothelial cell cultures were grown were monitored. Below is a
table summarizing the data.
4 Current Current Current Density Density Density Well Day 1 Day 2
Day 3 Num- (mA/ (mA/ (mA/ ber mm.sup.2) mm.sup.2) mm.sup.2)
Observations A1 2.10 1.99 PULLED Zone of death on the near edge of
OUT the well closest to the outside electrode A2 1.66 1.55 PULLED
Zone of death on the near edge of OUT the well closest to the
outside electrode A3 1.77 1.66 PULLED Zone of death on the near
edge of OUT the well closest to the outside electrode A4 0.99 0.99
PULLED Some floating cells, OUT No significant cell death near
outside electrode A5 0.88 0.88 PULLED Some floating cells, OUT No
significant cell death near outside electrode A6 0.00 0.00 PULLED
Some floating cells, OUT Cells appear to be healthy B1 2.10 2.21
2.24 Zone of death on the near edge of the well closest to the
outside electrode B2 1.35 1.33 1.55 Zone of death on the near edge
of the well closest to the outside electrode B3 1.55 1.77 1.88 Zone
of death on the near edge of the well closest to the outside
electrode B4 0.84 0.88 1.11 Starting to see cell death around the
edge of the well closest to the outside electrode B5 0.84 0.88 0.94
Starting to see cell death around the edge of the well closest to
the outside electrode B6 0.00 0.00 0.00 Some floating cells, Cells
appear to be healthy
[0096] By looking at the data shown above, 0.9-1.0 mA/mm.sup.2
appears to be the current threshold for damaging the endothelial
cells with in vitro electrical stimulation.
[0097] Cell Culture Studies--Example 1
[0098] Human coronary artery endothelial cells (Clonetics, San
Diego, Calif.) were seeded in 24 well tissue culture plates at
1.times.10.sup.4 cells/cm.sup.2 (1.5 mL total volume) and grown in
defined media as supplied by Clonetics. The cells were incubated at
37.degree. C., 5% CO.sub.2 for up to four days. The media was not
changed on the cells during the course of the experiment.
Supernatants from duplicate wells were withdrawn on days 2, 3, and
4 post-seeding. In addition, cells plus supernatants from a second
set of samples were harvested from duplicate wells on days 2, 3,
and 4 post-seeding. Samples were stored at -85.degree. C. until
assayed. The ELISA assay was performed on the test samples and on
controls following the procedure outlined by the manufacturer
(American Diagnostica, Inc.) The standard curve was generated using
control plasma A, t-PA antigen control plasma set 2 (Product #275,
American Diagnostica, Inc.). The test samples were assayed
undiluted (20 .mu.L assay volume). The results suggest that
measurable t-PA was released from control endothelial cells, in the
absence of stimulation. The levels of t-PA that were released were
correlated to cell growth (more was measured after 4 days of
culture than after 2 days).
[0099] Cell Culture Studies--Example 2
[0100] This experiment was performed similarly to the first
experiment with the following changes. In the first part of the
experiment, cells were seeded at three different densities
(1.times.10.sup.4, 2.times.10.sup.4, 4.times.10.sup.4) in 0.5 mL
total volume per well, and the cell supernatants were collected on
day 2 and day 3 post seeding. In the second part of the experiment,
cells were seeded at 2.times.10.sup.4 cells/cm.sup.2 and one group
was stimulated once (on day 2 post seeding) and a second group was
stimulated two times (once on day 2 and once on day 3 post
seeding). The stimulation level was 55V (350-440 mA) for 5 minutes
and supernatants were collected 15 minutes after stimulation. The
following tables contain the data that was obtained from this
experiment. The data confirms what was observed in the first
experiment--that t-PA production/release is correlated with cell
number and with cell growth. Although this data from the stimulated
samples suggests that electrical stimulation at 55 volts for 5
minutes did not increase t-PA production/release as compared to the
unstimulated control, it did not cause adverse effects on the
released t-PA.
5 Density Day 2 Day 3 st dev std 1 .times. 10.sup.4 15.153 23.208
0.088 1.73 2 .times. 10.sup.4 33.164 41.219 5.9 1.566 4 .times.
10.sup.4 44.773 54.926 3.309 1.755 Condition Day 1 Day 2 st dev std
No Stim 8.581 12.447 0.366 0.278 1 .times. Stim 8.08 10.86 0.54
0.176 2 .times. Stim 11.264 0.663
[0101] The complete disclosures of the patents, patent
applications, and publications listed herein are incorporated by
reference, as if each were individually incorporated by reference.
The above examples and disclosure are intended to be illustrative
and not exhaustive. These examples and description will suggest
many variations and alternatives to one of ordinary skill in this
art. All these alternatives and variations are intended to be
included within the scope of the attached claims. Those familiar
with the art may recognize other equivalents to the specific
embodiments described herein which equivalents are also intended to
be encompassed by the claims attached hereto.
* * * * *