U.S. patent application number 17/022855 was filed with the patent office on 2021-02-25 for methods for better delivery of active agents to tumors.
The applicant listed for this patent is SORRENTO THERAPEUTICS, INC.. Invention is credited to Russell F. Ross.
Application Number | 20210052871 17/022855 |
Document ID | / |
Family ID | 1000005210021 |
Filed Date | 2021-02-25 |
View All Diagrams
United States Patent
Application |
20210052871 |
Kind Code |
A1 |
Ross; Russell F. |
February 25, 2021 |
METHODS FOR BETTER DELIVERY OF ACTIVE AGENTS TO TUMORS
Abstract
The present invention concerns delivery of agents through the
skin. Methods for delivering agents such as bioactive agents are
contemplated by the present invention. Specifically, methods for
the targeted delivery of agents to one or more areas of the
epidermis and thereby, to one or more cancer tumors are
described.
Inventors: |
Ross; Russell F.;
(Jacksonville Beach, FL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SORRENTO THERAPEUTICS, INC. |
San Diego |
CA |
US |
|
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Family ID: |
1000005210021 |
Appl. No.: |
17/022855 |
Filed: |
September 16, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15744346 |
Jan 12, 2018 |
10806913 |
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PCT/US2016/043623 |
Jul 22, 2016 |
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17022855 |
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62196570 |
Jul 24, 2015 |
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62196578 |
Jul 24, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61M 2037/0061 20130101;
A61M 2037/0023 20130101; A61K 9/0014 20130101; A61M 2037/0038
20130101; A61M 2037/0007 20130101; A61K 9/0021 20130101; A61M
37/0015 20130101 |
International
Class: |
A61M 37/00 20060101
A61M037/00; A61K 9/00 20060101 A61K009/00 |
Claims
1. A method of treating a subject with a disease comprising one or
more tumors by administering one or more bioactive agents to the
one or more tumors comprising: (a) applying one or more delivery
devices having between 2 and 50,000 delivery structures to one or
more sites of a skin of a subject comprising blood vasculature and
lymphatic vasculature, wherein the delivery device contacts one or
more layers of epidermis with one or more reversible permeability
enhancers that induces a reversible increase in the permeability of
one or more barrier cells of the epidermis to at least the one or
more bioactive agents; (b) administering a total liquid dosage in
between 2 and 50,000 sub-doses of the one or more bioactive agents
at a controlled administration flow rate through the delivery
device; wherein each sub-dose of the one or more bioactive agents
is independently administered, in an administering step, to a
plurality of independent depths within the epidermis prior to any
subsequent diffusion or movement of the one or more bioactive
agents within the epidermis; and wherein following the
administering step, the one or more bioactive agents moves or
diffuses deeper through the epidermis through a basal layer of the
epidermis and into at least a portion of underlying viable dermis
to achieve an uptake of a portion of the one or more bioactive
agents by one or more susceptible blood capillary plexus or
lymphatic capillary plexus; wherein after administration and
uptake, the one or more bioactive agents circulates through the
blood vasculature or lymphatic vasculature to one or more tumors;
and wherein a greater concentration of the one or more bioactive
agents is delivered to the one or more tumors compared to
intravenous, intradermal, or subcutaneous delivery of the identical
one or more bioactive agents.
2. The method according to claim 1, wherein the total liquid dosage
of the one or more bioactive agents administered to the plurality
of independent depths within the epidermis comprises administration
to a depth within at least a portion of non-viable epidermis and/or
at least a portion of viable epidermis.
3. The method according to claim 1, wherein the plurality of
independent depths within the epidermis is from about 1 .mu.m to
about 500 .mu.m beyond a most superficial surface layer of the
epidermis of the subject.
4. The method according to claim 1, wherein the total liquid dosage
of the one or more bioactive agents is administered to a plurality
of depths within the epidermis consisting only of one or more
viable epidermal layers and not a non-viable epidermal layer.
5. The method according to claim 4, wherein the plurality of depths
within the viable epidermis is from about 1 .mu.m to about 250
.mu.m beyond the deepest non-viable epidermal layer but still
within the viable epidermis.
6. The method according to claim 1, wherein the average of the
plurality of independent depths exhibits a combined average
sub-dose delivery depth within the epidermis of about 70 .mu.m to
about 175 .mu.m beyond the most superficial surface layer of the
epidermis.
7. The method according to claim 1, wherein the plurality of
independent depths has a combined average depth of administration
within the epidermis, wherein each independently administered
sub-dose is at a depth within the epidermis that is deeper,
shallower, or the same.
8. The method according to claim 1, wherein a frequency of each of
the independent sub-dose administration depth within the viable
and/or non-viable epidermis exhibits a Gaussian distribution of
depths.
9. The method according to claim 1, wherein the delivery device
comprises an array comprising between 2 and 50,000 of the delivery
structures in fluid communication with the one or more bioactive
agents in a liquid carrier vehicle, wherein the delivery device
comprises a means for controlling the administration flow rate
including at least one component selected from the group consisting
of a pump, a fluid delivery rate controller, a syringe, a pen, an
elastomer membrane, or any combination thereof; wherein the
delivery structures comprise a means for penetrating at least a
most superficial layer of the epidermis; and wherein the one or
more bioactive agents in the liquid carrier vehicle is delivered by
the delivery structures to the plurality of independent depths
within a viable epidermis of the subject, thereby administering
between 2 and 50,000 sub-doses of the one or more bioactive
agents.
10. The method according to claim 9, wherein the one or more
bioactive agents is administered at a controlled administration
flow rate of about 0.01 .mu.l/hr to about 100 .mu.l/hr per delivery
structure.
11. The method according to claim 9, wherein the delivery
structures comprise a standard or nonstandard geometric shape.
12. The method according to claim 1, wherein the overall controlled
administration flow rate of the one or more bioactive agents to the
plurality of depths within the epidermis is from about 0.02
.mu.l/hr/cm2 to about 50,000 .mu.l/hr/cm2 based on the total
surface area of a delivery device that is in contact with the skin
of the subject.
13. The method according to claim 1, wherein the one or more
permeability enhancers is one or more chemical, physical, or
electrical permeability enhancers.
14. The method according to claim 13, wherein the physical
permeability enhancers comprises a nanostructured or nanotopography
surface.
15. The method according to claim 1, wherein the delivery
structures comprise needles.
16. The method according to claim 1, wherein the one or more
bioactive agents is delivered to a tissue volume of the epidermis
encompassing the one or more bioactive agents prior to any
subsequent diffusion or movement of the one or more bioactive
agents within the epidermis of about 0.7 mm3 to about 2,500
mm3.
17. The method according to claim 1, wherein administration of one
or more bioactive agents achieves a dermal interstitial fluid
pressure in the portion of underlying viable dermis beneath a site
of administration of about 1 mmHg to about 15 mmHg.
18. The method according to claim 1, wherein a concentration of the
one or more bioactive agents within the one or more tumors is about
1.25 fold to about 50 fold more than intravenous, intradermal, or
subcutaneous delivery of the one or more bioactive agents.
19. The method according to claim 1, wherein the bioactive agent is
useful for retarding progression of, delaying onset of, prophylaxis
of, amelioration of or reducing symptoms of the disease comprising
the one or more tumors.
20. The method according to claim 1, wherein the one or more
bioactive agents is continuously administered to a subject for a
time period of about 0.1 hours to about 96 hours.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of application Ser. No.
15/744,346, filed Jan. 12, 2018, which is a United States National
Phase Application of PCT/US2016/43623, filed Jul. 22, 2016, which
claims priority to U.S. Provisional Patent Applications 62/196,570
and 62/196,578 both having a filing date of Jul. 24, 2015, all of
which are incorporated by reference in their entirety.
TECHNICAL FIELD
[0002] The present invention concerns delivery of agents through
the skin. Methods for delivering agents such as bioactive agents
are contemplated by the present invention. Specifically, methods
for the targeted delivery of agents to one or more areas of the
epidermis and thereby, to one or more cancer tumors are
described.
BACKGROUND
[0003] Cancer is the second leading cause of mortality in the
United States, superseded only by heart disease with solid tumors
accounting for more than 85% of cancer mortalities. Currently, the
standard of care treatment for patients presenting with solid
tumors is invasive surgery followed by adjuvant chemotherapy and/or
radiotherapy. While this strategy has been successfully employed at
times, it is accompanied with cytotoxicity to normal cells and
tissues, in addition to the development of multidrug resistance
(MDR).
[0004] Targeted cancer therapies offer the potential to improve the
treatment of solid tumors. The thought bas been by targeting
therapeutic agents to solid tumors, cytotoxicity to normal cells
and tissues may be minimized and potentially limit the emergence of
drug resistance.
[0005] Current targeted delivery approaches that have been explored
include using nanoparticles (NPs), such as micelles, liposomes, and
dendrimers administered intravenously (i.v.) carrying a drug
payload for the targeted delivery of therapeutic agents to solid
tumors. Currently, systemic delivery of therapeutic agents via
nanoparticles to solid tumors is a three step process: (1) systemic
delivery of the therapeutic agent to different regions of the
tumor; (2) transport of the therapeutic agent across the vessel
wall into the solid tumor (extravasation); and (3) passage of the
therapeutic agent from the tumor tissue adjacent to the vasculature
to the tumor cells via diffusion through the interstitial
space.
[0006] Nanoparticles injected i.v. must remain in the systemic
circulation long enough for a portion to extravasate and accumulate
within a solid tumor tissue. Nanoparticles are capable of
accumulating in solid tumors due to the enhanced permeability and
retention (EPR) effect Masumura, et al., Cancer Research, (46),
6387-6392 (1986). The EPR effect is a consequence of the abnormal
vasculature frequently associated with solid tumors. The
vasculature of tumors is typically characterized by blood vessels
containing poorly-aligned defective endothelial cells with wide
fenestrations often lacking smooth muscle and a basement membrane.
However, the extent of the presence of intra tumor vasculature,
high tumor interstitial tissue fluid pressure, and tumor
vasculature composition heterogeneity make consistent delivery
using these types of approaches problematic.
[0007] Thus, despite the presence of the EPR effect, these prior
approaches are severely limited as the majority of nanoparticles
(>95%) accumulate in other organs and tissues (e.g., the liver,
spleen, and lungs). Further accounting for this effect, is evidence
suggesting that larger nanoparticles more effectively accumulate
within tumors, but are subject to higher rates of clearance from
the blood circulation see, for example, Moghimim et al.,
Pharmacological Reviews, 2(53), 283-318 (2001).
[0008] Additional approaches have been to utilize specific
ligand/receptor interactions for an active targeting of drugs or
drug carrier nanoparticles or modifications to increase plasma
half-life to increase chances of the EPR effect. For example,
PEGylated drug carriers have been shown to have increased systemic
circulation retention. Modest increases in tumor delivery were
observed, but still >90% of the delivered dose was systemically
cleared within a few hours. Active targeting approaches may provide
increased drug release selectivity but are similarly limited as
they also rely on initial i.v. administration and subsequent
extravasation of the drug or drug carrier, which can similarly lead
to accumulation in distant tissues far from the tumor to be
treated.
[0009] For example, two nanoparticle drug formulations have been
approved by the FDA, DOXIL.RTM. (a 100 nm PEGylated liposomal form
of doxorubicin) and ABRAXANE.RTM. (an 130 nm albumin-bound
paclitaxel nanoparticle). While these formulations have exhibited
some improved pharmacokinetic properties and reduced adverse
effects, they provided only modest survival benefits. Thus, the
limited efficacy of these existing nanoparticle formulations likely
stems from their inability to effectively deliver the therapeutic
agents to the solid tumor.
[0010] Therefore new methods for delivering increased
concentrations of agents to solid tumors are greatly needed.
SUMMARY
[0011] One embodiment described herein is a method of delivering
one or more agents to one or more susceptible tumors of a subject,
the method comprising: (a) contacting one or more layers of
epidermis with one or more reversible permeability enhancers,
wherein the one or more reversible permeability enhancers induces a
reversible increase in the permeability of one or more barrier
cells of the epidermis to at least the one or more agents; (b)
administering a total liquid dosage in between 2 and 50,000
sub-doses of the one or more agents at a controlled administration
flow rate, wherein each sub-dose of the one or more agents is
independently administered to a plurality of independent depths
within the epidermis prior to any subsequent diffusion or movement
of the one or more agents within the epidermis; and wherein
following administration, the permeability of the one or more
barrier cells returns to a normal state prior to the contacting of
the epidermis with the one or more permeability enhancers.
[0012] Another embodiment described herein is a method of treating
a subject with a disease comprising one or more tumors by
administering one or more bioactive agents to the one or more
tumors comprising: (a) applying one or more delivery devices having
between 2 and 50,000 delivery structures to one or more sites of
skin comprising blood vasculature and lymphatic vasculature,
wherein the delivery device contacts one or more layers of
epidermis with one or more reversible permeability enhancers that
induces a reversible increase in the permeability of one or more
barrier cells of the epidermis to at least the one or more
bioactive agents; (b) administering a total liquid dosage in
between 2 and 50,000 sub-doses of the one or more bioactive agents
at a controlled administration flow rate through the delivery
device wherein each sub-dose of the one or more bioactive agents is
independently administered to a plurality of independent depths
within the epidermis prior to any subsequent diffusion or movement
of the one or more bioactive agents within the epidermis; wherein
following the administering step, the one or more bioactive agents
moves or diffuses deeper through the epidermis through a basal
layer of the epidermis and into at least a portion of underlying
viable dermis to achieve an uptake of a portion of the one or more
bioactive agents by one or more susceptible blood capillary plexus
or lymphatic capillary plexus; wherein after administration and
uptake, the one or more bioactive agents circulates through the
blood vasculature or lymphatic vasculature to one or more tumors;
and wherein a greater concentration of the one or more bioactive
agents is delivered to the one or more tumors compared to
intravenous, intradermal, or subcutaneous delivery of the identical
one or more bioactive agents.
[0013] In some aspects of the embodiments described herein, the
epidermis comprises both nonviable epidermis and viable
epidermis.
[0014] In some aspects of the embodiments described herein, the
plurality of independent depths has a combined average depth of
administration within the epidermis, wherein each independently
administered sub-dose is at a depth within the epidermis that is a
deeper depth, a shallower depth, or a same depth.
[0015] In some aspects of the embodiments described herein, the
total liquid dosage of the one or more agents administered to
plurality of depths within the epidermis comprises administration
to a depth within at least a portion of non-viable epidermis and/or
at least a portion of viable epidermis.
[0016] In some aspects of the embodiments described herein, the
plurality of depths within the epidermis is from about 1 .mu.m to
about 500 .mu.m beyond a most superficial surface layer of the
epidermis of the subject.
[0017] In some aspects of the embodiments described herein, the
total liquid dosage of the one or more agents is administered to a
plurality of depths within the epidermis consisting only of one or
more viable epidermal layers and not a non-viable epidermal
layer.
[0018] In some aspects of the embodiments described herein, the
plurality of depths within the viable epidermis is from about 1
.mu.m to about 250 .mu.m beyond the deepest non-viable epidermal
layer but still within the viable epidermis.
[0019] In some aspects of the embodiments described herein, the
average of the independent plurality of depths exhibits a combined
average sub-dose delivery depth within the epidermis of about 70
.mu.m to about 175 .mu.m beyond the most superficial surface layer
of the epidermis.
[0020] In some aspects of the embodiments described herein, a
frequency of each of the independent sub-dose administration depth
within the viable and/or non-viable epidermis exhibits a Gaussian
distribution of depths.
[0021] In some aspects of the embodiments described herein, the one
or more agents are administered by applying one or more delivery
devices to one or more sites of the skin.
[0022] In some aspects of the embodiments described herein, the
delivery device comprises an array comprising between 2 and 50,000
delivery structures in fluid communication with one or more agents
in a liquid carrier vehicle, wherein the delivery device comprises
a means for controlling the administration flow rate; wherein the
delivery structures comprise a means for penetrating at least a
most superficial layer of the epidermis; and wherein the one or
more agents in a liquid carrier vehicle is delivered by the
delivery structures to the plurality of depths within the viable
epidermis of a subject, thereby administering between 2 and 50,000
sub-doses of the one or more agents.
[0023] In some aspects of the embodiments described herein, the
delivery structures comprise a standard or non-standard geometric
shape.
[0024] In some aspects of the embodiments described herein, the
delivery structures comprise needles.
[0025] In some aspects of the embodiments described herein, the one
or more agents is administered at a controlled administration flow
rate of about 0.01 .mu.l/hr to about 100 .mu.l/hr per delivery
structure.
[0026] In some aspects of the embodiments described herein, the
overall controlled administration flow rate of the one or more
agents to the plurality of depths within the epidermis is from
about 0.02 .mu.l/hr/cm.sup.2 to about 50,000 .mu.l/hr/cm.sup.2
based on the total surface area of a delivery device that is in
contact with the skin of the subject.
[0027] In some aspects of the embodiments described herein, the one
or more agents is delivered to a tissue volume of the epidermis
encompassing the one or more agents prior to any subsequent
diffusion or movement of the one or more agents within the
epidermis of about 0.7 mm.sup.3 to about 2,500 mm.sup.3.
[0028] In some aspects of the embodiments described herein, the one
or more agents are continuously administered to a subject for a
time period of about 0.1 hours to about 96 hours.
[0029] In some aspects of the embodiments described herein, the one
or more permeability enhancers are one or more chemical, physical,
or electrical permeability enhancers.
[0030] In some aspects of the embodiments described herein, the
physical permeability enhancers comprise a nanostructured or
nanotopography surface.
[0031] In some aspects of the embodiments described herein, the
nanotopograhy surface is fabricated on the surface of the delivery
structures as described herein.
[0032] In some aspects of the embodiments described herein, the
administered one or more agents to the plurality of depths within
the skin moves or diffuses deeper through the epidermis through a
basal layer of the epidermis and into at least a portion of
underlying viable dermis.
[0033] In some aspects of the embodiments described herein, the
administration of one or more agents achieves a dermal interstitial
fluid pressure in the underlying dermis of about 1 mmHg to about 15
mmHg.
[0034] In some aspects of the embodiments described herein, the one
or more agents is absorbed by one or more tissues comprising one or
more susceptible lymphatic capillary plexus or one or more blood
capillary plexus following delivery to the epidermis.
[0035] In some aspects of the embodiments described herein, the one
or more agents circulate through the one or more blood capillary
plexus and into or within proximity to one or more susceptible
tumors.
[0036] In some aspects of the embodiments described herein, the one
or more agents circulate through the one or more lymphatic
capillary plexus and into or within proximity to one or more
susceptible tumors.
[0037] In some aspects of the embodiments described herein, the
concentration of one or more agents within one or more susceptible
tumors is about 1.25 fold to about 50 fold more than intravenous,
intradermal, or subcutaneous delivery of the identical one or more
agents.
[0038] In some aspects of the embodiments described herein, a blood
serum absorption rate of the one or more agents is equivalent to
intradermal delivery and subcutaneous delivery of the identical one
or more agents.
[0039] In some aspects of the embodiments described herein, the one
or more agents comprise a bioactive agent.
[0040] In some aspects of the embodiments described herein, the
bioactive agent is useful for treating, retarding the progression
of, delaying the onset of, prophylaxis of, amelioration of, or
reducing the symptoms of a disease in a patient in need of
treatment thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041] FIG. 1. Schematic of the skin including the epidermis and
dermis illustrating the various tissues of the skin.
[0042] FIGS. 2A and 2B. Schematic of the epidermis illustrating the
layers of the epidermis.
[0043] FIG. 3. Schematic of an exemplary delivery structure for
administering an active agent to the skin.
[0044] FIG. 4. Schematic of an exemplary delivery structure having
a nanotopography surface for administering an active agent to the
skin.
[0045] FIGS. 5A, 5B, and 5C. Schematic of the delivery methods to
the skin illustrating average depth of delivery.
[0046] FIG. 6. Optical coherence tomography (OCT) imaging of the
skin following penetration of the skin with a needle array.
[0047] FIGS. 7A, 7B, and 7C. Modulation of tight junction proteins
by nanotopography containing needles in Caco-2 epithelial
cells.
[0048] FIG. 8. Targeted delivery of an anti-cancer drug
(trastuzumab) to tumors in vivo following delivery to the skin of
rats.
[0049] FIGS. 9A and 9B Representative tumors following delivery of
an anti-cancer drug (trastuzumab) to the skin of rats.
[0050] FIGS. 10A and 10B. Distribution of drugs delivered to in
vitro grown tumors using an array of needles or by drug
supplemented tissue culture media.
[0051] FIGS. 10C and 10D. Drug delivery method effects on
proliferation of tumor tissue cells grown in vitro following drug
delivery using an array of needles or by drug supplemented tissue
culture media.
[0052] FIG. 11. Imaging of fluorescently tagged drug (Etanercept)
administered to the skin showing delivery directly to the lymphatic
vasculature and lymph node tissues.
[0053] FIG. 12. Imaging of fluorescently tagged drug (Etanercept)
administered to the skin showing delivery to the axillary and
mandibular lymph nodes.
[0054] FIG. 13. Biodistribution for Etanercept following delivery
to the skin using the methods of the present invention illustrating
increased delivery to the lymph nodes compared to traditional
delivery methods.
[0055] FIG. 14. Blood serum absorption rate of a drug (Etanercept)
following delivery to the skin showing that the blood serum
absorption rate is similar to traditional reference delivery
methods.
DETAILED DESCRIPTION
[0056] There is a need for methods of controlled delivery of agents
(e.g., bioactive agents) to solid cancer tumors of subjects.
Therefore, described herein are methods for the controlled delivery
of one or more agents to the skin followed by the uptake of the one
or more agents by tumors. In some embodiments described herein the
uptake of the one or more agents by one or more tumors is
facilitated by primary absorption of the lymphatic tissues followed
by delivery through the lymphatic vasculature to one or more
susceptible tumors.
[0057] The term "viable skin" as used herein refers to an area of
the skin immediately below the stratum corneum layer of the
epidermis including the dermis, but above the subcutaneous tissue
layers. This term encompasses both the viable epidermis and the
viable dermis. The actual depth of the viable skin will vary
depending on location of the skin, age, and physiology of a given
subject. The term viable skin further specifies that this portion
of the skin comprises nucleated living cells, often mitotic. In
some aspects described herein, the viable skin also comprises at
least one or more lymphatic capillary plexus and/or one or more
blood capillary plexus.
[0058] The term "viable dermis" as used herein refers to an area of
the skin immediately below the basal layer of the epidermis but
above the subcutaneous tissue layer. The viable dermis comprises
both the papillary and reticular dermal layers of the dermis,
further comprising, for example, blood capillaries and lymphatic
capillaries amongst other tissue types.
[0059] The term "viable epidermis" as used herein refers to an area
of the skin immediately below the stratum corneum. The viable
epidermis comprises the basal layer or stratum germinativum, the
squamous cell layer or the stratum spinosum and the granular cell
layer or the stratum granulosum.
[0060] The term "agent" as used herein refers to a compound,
substance, composition, or molecule to be delivered. Exemplary and
non-limiting examples include bioactive agents, nucleic acids
(e.g., micro RNAs), dyes (e.g., contrast agents and fluorescent
reporters), vaccines and the like.
[0061] The term "bioactive agent," as used herein refers to any
biocompatible agent, which elicits a cellular response. The term
bioactive agent comprises any drug, active ingredient, active drug
substance, or vaccine. For example, a bioactive agent described in
the embodiments herein, may comprise drugs, such as small molecule
drugs, bio-similar drugs, biologics, etc., nanoparticles, lipids,
liposomes, proteins (e.g., recombinant proteins, antibodies, etc.),
and the like.
[0062] The terms "drug", "active ingredient," "active drug
substance," or "active pharmaceutical agent" as used herein refer
to an active ingredient, compound, or substance, compositions, or
mixtures thereof, that provide a pharmacological, often beneficial,
effect. Reference to a specific active ingredient includes, where
appropriate, the active ingredient and any of its pharmaceutically
acceptable salts or esters.
[0063] The terms "dosage" or "dose" denote any form of the active
ingredient formulation that contains an amount sufficient to
produce a therapeutic effect with a single administration.
[0064] The term "titration" as used herein refers to the
incremental increase in drug dosage or administration rate to a
level that provides the optimal therapeutic effect.
[0065] The term "controlled delivery" as used herein refers to an
administration method that results in the controllable delivery of
one or more agents over a desired period of time. As used herein,
it encompasses the terms "modified delivery," "sustained delivery,"
"extended delivery," and "delayed delivery." In some aspects
described herein, the methods for controlled delivery result in the
delivery of one or more agents or active drug substances to achieve
a therapeutic threshold for a maximal length of time.
[0066] The term "delayed delivery" as used herein refers to the
delivery of one or more agents according to a desired profile over
an extended period under physiological conditions or in an in vitro
test. By "extended period" it is meant a continuous period of time
of at least about 20 minutes, about 30 minutes, about 1 hour; about
2 hours; about 4 hours; about 6 hours; about 8 hours; about 10
hours; about 12 hours; about 14 hours; about 16 hours; about 18
hours; about 20 hours about 24 hours; or even longer.
[0067] The term "modified delivery" as used herein refers to the
delivery of one or more agents at a slower rate than does immediate
delivery formulation under physiological conditions or in an in
vitro test.
[0068] The term "sustained delivery" as used herein refers to the
delivery of one or more agents over an extended period of time, for
example minutes, hours, or days, such that less than all the active
ingredient is released initially. A sustained release rate may
provide, for example, the delivery of a certain specified amount of
one or more agents or active drug substances over a certain period,
under physiological conditions or in an in vitro test.
[0069] The term "extended delivery" as used herein refers to the
delivery of one or more agents over an extended period, such as of
at least about 20 minutes, about 30 minutes, about 1 hour; about 2
hours; about 4 hours; about 6 hours; about 8 hours; about 10 hours;
about 12 hours; about 14 hours; about 16 hours; about 18 hours;
about 20 hours, about 24 hours, about 48 hours, about 72 hours; or
even longer.
[0070] The term "initial delivery" or "initially delivered" refers
to a tissue location at which an agent first comes into contact. In
some aspects described herein, initial delivery may refer to a
location within the skin (e.g., non-viable epidermis, viable
epidermis, or viable dermis) in which one or more agents first
contacts after being delivered through a delivery device or one or
more delivery structures of a delivery device.
[0071] As used herein, "conventional delivery" means any method
prior to the present invention that is used in the art for
delivering one or more materials having biological kinetics or
activity similar to intravenous (i.v.), iontophoretic, subcutaneous
(s.c.), intramuscular (i.m.), or intradermal (i.d.) injections, or
topical formulations. Exemplary methods include subcutaneous,
iontophoretic, and intradermal delivery methods, such as those
described in U.S. Pat. No. 5,800,420, US 20050180952, Xie et al.,
Expert Opin Drug Deliv., 6(8), 785-792 (2009) and Zhang and
Wei-Yue., Cancer Biol Med., (11), 247-254 (2014), each of which is
incorporated by reference herein with regard to a general
description of conventional delivery methods.
[0072] The term "targeted drug delivery" refers to the predominant
location, wherein a drug accumulates. This term is separate and
distinct from commonly used terminology, such as "targeted
therapy," which more specifically refers to a specific interaction
with a cell or tissue type (e.g., a ligand/receptor
interaction).
[0073] The term "BCS Class I, II, II, or IV" refers to whether a
compound or active drug substance has high or low permeability and
high or low solubility (e.g., poorly soluble). BCS Class I drugs
have high permeability and high solubility; BCS Class II drugs have
high permeability and low solubility, BCS Class III drugs have low
permeability and high solubility, and BCS Class IV drugs have low
permeability and low solubility. An immediate release drug
substance is considered highly soluble when the highest dose
strength is soluble in 250 mLs or less of aqueous media over the pH
range of 1 to 7.5 at 37.+-.1.degree. C. A sufficient number of pH
conditions should be evaluated to accurately define the
pH-solubility profile. In the absence of evidence suggesting
instability in the gastrointestinal tract, an immediate release
drug substance is considered to be highly permeable when the extent
of absorption in humans is determined to be 90% or more of an
administered dose based on the mass balance determination or in
comparison to an intravenous reference dose. Permeability can be
determined using mass balance, absolute bioavailability, or
intestinal perfusion approaches. When a single method fails to
conclusively demonstrate the permeability classification, two
different methods may be advisable. A drug product is considered
rapidly dissolving when no less than 85% of the labeled amount of
the drug substance dissolves within 30 minutes, using USP Apparatus
I at 100 rpm (or Apparatus II at 50 rpm) in a volume of 900 ml or
less in each of the following media: (1) 0.1 N HCl or Simulated
Gastric Fluid USP without enzymes; (2) a pH 4.5 buffer; and (3) a
pH 6.8 buffer or Simulated Intestinal Fluid USP without enzymes.
See, FDA Guidance for Industry: Waiver of In Vivo Bioavailability
and Bioequivalence Studies for Immediate-Release Solid Oral Dosage
Forms Based on a Biopharmaceutics Classification System. (August
2000), which is incorporated by reference herein for such
teachings.
[0074] As used herein, "bioavailability", means the total amount of
a given dosage of the administered agent that reaches the blood
compartment. This is generally measured as the area under the curve
in a plot of concentration vs. time.
[0075] As used herein "tissue" refers to a group or layer of cells
that together perform a function including but not limited to, skin
tissue, lymphatic tissue (e.g., lymph nodes), mucosal tissue,
reproductive tissue, cervical tissue, vaginal tissue and any part
of the body that consists of different types of tissue and that
performs a particular function, i.e., an organ, including but not
limited to lung, spleen, colon, thymus. As used herein, tissue
includes any tissue that interacts with or is accessible to the
environment, e.g., skin or mucosal tissue.
[0076] As used herein, "tissue-bioavailability" means the amount of
an agent that is biologically available in vivo in a particular
tissue. These amounts are commonly measured as activities that may
relate to binding, labeling, detection, transport, stability,
biological effect, or other measurable properties useful for
diagnosis and/or therapy. In addition, it is understood that the
definition of "tissue-bioavailability" also includes the amount of
an agent available for use in a particular tissue.
"Tissue-bioavailability" includes the total amount of the agent
accumulated in a particular tissue, the amount of the agent
presented to the particular tissue, the amount of the agent
accumulated per mass/volume of particular tissue, and amount of the
agent accumulated per unit time in a particular mass/volume of the
particular tissue. Tissue bioavailability includes the amount of an
agent that is available in vivo in a particular tissue or a
collection of tissues such as those that make up the vasculature
and/or various organs of the body e.g., a part of the body that
consists of different types of tissue and that performs a
particular function.
[0077] The term "C.sub.max" as used herein refers to the maximum
observed blood (plasma, serum, or whole blood) concentration or the
maximum blood concentration calculated or estimated from a
concentration to time curve, and is expressed in units of mg/L or
ng/mL, as applicable.
[0078] The term "C.sub.min" as used herein refers to the minimum
observed blood (plasma, serum, or whole blood) concentration or the
minimum blood concentration calculated or estimated from a
concentration to time curve, and is expressed in units of mg/L or
ng/mL, as applicable.
[0079] The term "C.sub.avg" as used herein refers to the blood
(plasma, serum, or whole blood) concentration of the drug within
the dosing interval, is calculated as AUC/dosing interval, and is
expressed in units of mg/L or ng/mL, as applicable.
[0080] The term "T.sub.max" as used herein refers to the time after
administration at which C.sub.max occurs, and is expressed in units
of hours (h) or minutes (min), as applicable.
[0081] The term "AUC.sub.0.fwdarw..tau." as used herein refers to
area under the blood (plasma, serum, or whole blood) concentration
versus time curve from time zero to time tau (.tau.) over a dosing
interval at steady state, where tau is the length of the dosing
interval, and is expressed in units of hmg/L or hng/mL, as
applicable. For example, the term AUC.sub.0.fwdarw.12 as used
herein refers to the area under the concentration versus time curve
from 0 to 12 hours.
[0082] The term "AUC.sub.0.fwdarw..infin." as used herein refers to
the area under the blood (plasma, serum, or whole blood)
concentration versus time curve from time 0 hours to infinity, and
is expressed in units of h mg/L or hng/mL, as applicable.
[0083] The term "AUC.sub.overall" as used herein refers to the
combined area under the blood (plasma, serum, or whole blood)
concentration versus time curve, and is expressed in units of h
mg/L (or h ng/mL) for at least one or more doses of the
pharmaceutical compositions described herein. In one aspect, the
"AUC.sub.overall" refers to the combined area under the blood
concentration versus time curve for at least two doses of the
pharmaceutical compositions described herein.
[0084] The term "treating" refers to administering a therapy in an
amount, manner, or mode effective to improve a condition, symptom,
or parameter associated with a disorder.
[0085] The term "prophylaxis" refers to preventing or reducing the
progression of a disorder, either to a statistically significant
degree or to a degree detectable to one skilled in the art.
[0086] The term "substantially" as used herein means to a great or
significant extent, but not completely. In some aspects,
substantially means 90% to 99% or more in the various embodiments
described herein, including each integer within the specified
range.
[0087] As used herein, the terms "subject" and "patient" are used
interchangeably. As used herein, a subject is preferably a mammal
such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats
etc.) and a primate (e.g., monkey and human), most preferably a
human.
[0088] As used herein, the terms "disorder" and "disease" are used
interchangeably to refer to a condition in a subject. Diseases
include to any interruption, cessation, or disorder of body
functions, systems or organs.
[0089] As used herein, the terms "treat," "treating" and
"treatment" refer to the eradication, reduction or amelioration of
symptoms of a disease or disorder. In some embodiments, treatment
refers to the eradication, removal, modification, or control of
primary, regional, or metastatic cancer tissue that result from the
administration of one or more therapeutic agents. In certain
embodiments, such terms refer to the minimizing or delaying the
spread of cancer resulting from the administration of one or more
therapeutic agents to a subject with such a disease.
[0090] As used herein, the terms "manage," "managing" and
"management" refer to the beneficial effects that a subject derives
from administration of a prophylactic or therapeutic agent, which
does not result in a cure of the disease. In certain embodiments, a
subject is administered one or more prophylactic or therapeutic
agents to "manage" a disease so as to prevent the progression or
worsening of the disease.
[0091] As used herein, the terms "prevent", "preventing" and
"prevention" refer to the prevention of the recurrence or onset of
one or more symptoms of a disorder in a subject resulting from the
administration of a prophylactic or therapeutic agent
[0092] As used herein, the phrase "side effects" encompasses
unwanted and adverse effects of a prophylactic or therapeutic
agent. Adverse effects are always unwanted, but unwanted effects
are not necessarily adverse. An adverse effect from a prophylactic
or therapeutic agent might be harmful or uncomfortable or risky.
Side effects from chemotherapy include, but are not limited to,
gastrointestinal toxicity such as, but not limited to, early and
late-forming diarrhea and flatulence, nausea, vomiting, anorexia,
leukopenia, anemia, neutropenia, asthenia, abdominal cramping,
fever, pain, loss of body weight, dehydration, alopecia, dyspnea,
insomnia, dizziness, mucositis, xerostomia, and kidney failure, as
well as constipation, nerve and muscle effects, temporary or
permanent damage to kidneys and bladder, flu-like symptoms, fluid
retention, and temporary or permanent infertility. Side effects
from radiation therapy include but are not limited to fatigue, dry
mouth, and loss of appetite. Side effects from biological
therapies/immunotherapies include but are not limited to rashes or
swellings at the site of administration, flu-like symptoms such as
fever, chills and fatigue, digestive tract problems and allergic
reactions. Side effects from hormonal therapies include but are not
limited to nausea, fertility problems, depression, loss of
appetite, eye problems, headache, and weight fluctuation.
Additional undesired effects typically experienced by patients are
numerous and known in the art, see, e.g., the Physicians' Desk
Reference (69.sup.th ed., 2015), which is incorporated herein by
reference in its entirety.
[0093] As used herein, the phrase "delivery to a susceptible
tissue" or a "viable tissue" refers to the delivery of one or more
agents to a living tissue or tissue structure, for example the
skin, spleen, thymus, lung, vasculature, lymphatic vasculature,
lymph nodes, heart and brain, etc. In some embodiments described
herein, the methods, compositions, and devices further described
herein may modulate the structure of a living tissue or tissue
structure to facilitate the absorption of one or more agents. In
some aspects, the living tissue or tissue structure includes the
skin and individual viable cells that comprise the skin. In some
aspects, described herein, the methods of delivery induce a
particular cell or tissue (e.g., the viable skin) to be susceptible
to delivering one or more agents to that specific tissue. In some
aspects described herein, the living tissue or tissue structure
comprises one or more layers of the viable skin such as the viable
layers of the epidermis and the underlying dermis. In some aspects
described herein, the living tissue or tissue structure comprises
lymphatic capillaries, e.g., delivery to a susceptible lymphatic
capillary plexus.
[0094] Described herein are methods and devices for the initial
delivery of agents to the skin, and subsequently to a susceptible
tumor. In certain embodiments described herein are methods for
delivering agents to the lymphatic vasculature and a susceptible
tumor.
[0095] In some embodiments described herein are methods for
delivering one or more agents to the skin. In some aspects, the one
or more agents are delivered to at least a portion or area of the
viable skin or non-viable skin. In some aspects, the one or more
agents are delivered to at least a portion or area of the viable
epidermis. In some aspects, the one or more agents are delivered to
at least a portion or area of the non-viable epidermis. As further
described herein, the one or more agents are able to pass through
the viable epidermis and enter the dermis, thereby coming into
proximity with one or more blood or lymphatic capillaries. In some
embodiments described herein, delivery of one or more agents to the
skin results in the uptake of the one or more agents by a
susceptible tumor. The delivery to tumors may be due to absorption
by a lymphatic capillary or a blood capillary or both as further
described herein.
[0096] Delivery to the skin presents several difficulties based
upon the barrier providing function of the skin. Anatomically, the
skin is broadly made up of two major tissue layers, an outer
epidermis and an underlying dermis, which together constitute the
skin. The broader integumentary system comprises the skin, hair,
nails, exocrine glands, and the subcutaneous tissues. Many
transdermal or microneedle approaches for delivery to the skin and
through the epidermis and into the viable dermis are unsuccessful
because of this barrier function resulting in the delivered
materials being retained within one or more layers of the
epidermis.
[0097] The epidermis is subdivided into four principle layers or
strata. In order from bottom to top is the basement membrane, the
basal layer or stratum germinativum, the squamous cell layer or the
stratum spinosum, the granular cell layer or the stratum
granulosum, and the cornified layer or the stratum corneum. Of
these three layers, the lower three layers (i.e., stratum
germinativum, stratum spinosum, and stratum granulosum) constitute
the living layers of the epidermis.
[0098] These living layers of the epidermis are important for the
barrier function of the skin, which relies on the self-renewal and
differentiation of the basally located stem cells to regenerate the
upper layers of the skin and provide enucleated cells for the
barrier layer or the stratum corneum. The barrier function of the
epidermis is largely due to the presence of tight junctions which
prevent the passage of macromolecules (e.g., proteins),
microorganisms, and other potentially toxic chemicals. Thus, these
tight junctions are barrier structures that include a network of
transmembrane proteins embedded in adjacent plasma membranes (e.g.,
claudins, occludin, and junctional adhesion molecules) as well as
multiple plaque proteins (e.g., ZO-1, ZO-2, ZO-3, cingulin,
symplekin). Tight junctions are found in nearly all types of
barrier types of tissue including the internal epithelium (e.g.,
the intestinal epithelium, the blood-brain barrier, blood vessels,
lymphatic vessels) as well as throughout the viable epidermis of
the skin.
[0099] The thickness of the skin is varied depending on location
and age. For example the eye lid has one of the thinnest layers of
epidermis at less than about 0.2 mm; the palms of the hands and
soles of the feet have some of the thickest layers of epidermis
measuring at nearly 1.5 mm. The thickness of the dermis is also
varied depending on tissue location with the dermis on the back
being 30-40 times thicker than the epidermis see, William D. James,
Timothy Berger, and Dirk Elston., Clinical Dermatology (11.sup.th
ed. 2011), which is incorporated by reference herein in its
entirety.
[0100] Beneath the epidermis lies the dermis, which contains two
layers, an outermost portion referred to as the papillary dermis
and a deeper layer referred to as the reticular dermis. The
papillary dermis contains vast microcirculatory blood and lymphatic
plexuses. In contrast, the reticular dermis is relatively
acellular, made up of dense collagenous and elastic connective
tissue. Beneath the epidermis and dermis is the subcutaneous
tissue, also referred to as the hypodermis, which is composed of
connective tissue and fatty tissue. See, Physiology, Biochemistry,
and Molecular Biology of the Skin, Second Edition, (L. A.
Goldsmith, Ed., 2.sup.nd ed. Oxford University Press, New York,
1991), which is incorporated by reference herein in its
entirety.
[0101] Some embodiments described herein are methods for the
targeted delivery of one or more agents to one or more tumors. The
delivery of one or more agents to one or more tumors is facilitated
by the delivery of one or more agents to the skin at a rate and
depth as further described herein. The targeted delivery of one or
more agents to one or more tumors may be facilitated by delivery to
one or more susceptible lymphatic capillary plexus. In some other
aspects, the targeted delivery of one or more agents to one or more
tumors may be facilitated by the delivery to one or more
susceptible blood capillary plexus. In some aspects, the tumor may
be a primary tumor or a secondary tumor (e.g., a metastasis of the
primary tumor).
[0102] In some embodiments described herein, one or more agents are
delivered to a position within the skin, wherein after the initial
administration, the one or more agents moves or diffuses to a
position that is in proximity of the blood vasculature and the
lymphatic vasculature. As described herein, this placement within
the skin may result in the subsequent delivery of an agent to a
lymphatic capillary bed or otherwise known as a lymphatic drainage
bed or lymphatic capillary plexus, which physiologically functions
to drain interstitial fluid for a given location to the rest of the
lymphatic system.
[0103] In some embodiments described herein, one or more agents are
directly delivered to a position within the epidermis. In some
aspects, the one or more agents diffuse, move, flow, or migrate to
a position in proximity to the lymphatic vasculature. As described
herein, this placement within the epidermis following the methods
described herein results in the diffusion or movement of an agent
through the epidermis and into the viable epidermis, which allows
for direct contact of an agent to the most superficially present
lymphatic capillary bed(s) or otherwise known as a lymphatic
drainage bed or lymphatic capillary plexus, which physiologically
functions to drain interstitial fluid for a given location to the
rest of the lymphatic system. In some other aspects, this placement
within the skin may result in the localized delivery of an agent to
a blood capillary bed. The methods of delivering one or more agents
to a lymphatic capillary bed described herein may further result in
the delivery of the agent to the first lymph nodes draining the
lymphatic capillary bed, also referred to as "primary" lymph nodes.
In some aspects, the localized delivery of one or more agents may
also result in the delivery of the agent to additional lymph nodes
downstream of the primary lymph nodes, also referred to as
"secondary" lymph nodes. In some aspects the agent may eventually
enter the blood stream and be delivered systemically. In some
aspects described herein, the delivery of one or more agents to the
skin results in the targeted delivery of the one or more agents to
one or more susceptible tumors in a subject.
[0104] In some embodiments described herein are methods for
delivering one or more agents to a range of depths within the skin.
In some aspects, the one or more agents is delivered to the
epidermis, which comprises both the non-viable epidermis (e.g.,
stratum corneum) and the viable epidermis underlying the non-viable
epidermis. The depth in the skin may vary depending on location,
age and physiology of the skin of a given subject as described
herein. The overall depth in the skin of delivery of one or more
agents may be described as the distribution of a plurality of
depths that the one or more agents may be located following the
initial administration of the one or more agents using the methods
described herein. The total distribution of depths of delivery of
the one or more active agents depends on the rate of
administration, volume, and depth within the skin of a delivery
structure as described further herein. Therefore, portions of the
total delivered agent may be at a more superficial depth or a
deeper depth, wherein the total delivered agent has an average
delivery depth and standard deviation of a range of delivery
depths. Therefore, in some aspects, the delivery of one or more
agents to the skin as described herein may follow a simple normal
distribution (i.e., a Gaussian distribution) within the skin. In
some other aspects, the delivery of one or more agents to the skin
may follow a multi-modal distribution of depths within the
skin.
[0105] As further described herein, the delivery of one or more
agents to the epidermis, wherein the administered one or more
agents exhibits a distribution of depths within the epidermis
provides allows for increased lymphatic uptake of the one or more
agents. The delivery methods described herein allow for the
previously unrealized aspect of contacting all levels of potential
dermal lymphatic capillaries. The methods described herein further
comprise reversibly increasing the porosity of the barrier function
of the skin to promote the downward (top to bottom) diffusion or
movement of an agent throughout all layers of the epidermis and
into the viable dermis. In some aspects described herein, delivery
to the epidermis yields greater lymphatic uptake compared to
alternative parenteral delivery methods, such as direct intradermal
delivery techniques, which may miss the initial lymphatic
capillaries directly below the basement membrane of the epidermis,
resulting in reduced lymphatic uptake. Without being bound by any
theory, this may occur because the agent may more freely move
downwardly through the reticular dermis and into the subcutaneous
tissue. Therefore, by providing methods that allow for the
diffusion or movement of an agent through the epidermis at a
plurality of flow rates as described herein, the superficial
lymphatics and deeper lymphatics within the dermis may be contacted
by an agent, which increases the absorption rate or amount of an
agent by one or more susceptible lymphatic capillaries.
[0106] In some embodiments described herein, at least a portion of
or all of one or more agents may be directly delivered or
administered to an initial depth in the skin comprising the
nonviable epidermis and/or the viable epidermis. In some aspects, a
portion of the one or more agents may also be directly delivered to
the viable dermis in addition to the epidermis. The range of
delivery depth will depend on the disease being treated and the
skin physiology of a given subject. This initial depth of delivery
may be defined as a location within the skin, wherein an
administered agent first comes into contact as described herein.
Without being bound by any theory, it is thought that the
administered one or more agents may move (e.g., diffuse) from the
initial site of delivery (e.g., the non-viable epidermis, the
viable epidermis, or the viable dermis) to a deeper position within
the viable skin. For example, a portion of or all of an
administered agent may be delivered to the non-viable epidermis and
then continue to move (e.g., diffuse) into the viable epidermis and
past the basal layer of the viable epidermis and enter into the
viable dermis. Alternatively, a portion of or all of an
administered agent may be delivered to the viable epidermis (i.e.,
immediately below the stratum corneum) and then continue to move
(e.g., diffuse) past the basal layer of the viable epidermis and
enter into the viable dermis. Lastly, a portion of or all of an
administered agent may be delivered to the viable dermis. The
movement of the one or more active agents throughout the skin is
multifactorial and, for example, depends on the liquid carrier
composition (e.g., viscosity thereof), rate of administration,
delivery structures, etc. This movement through the epidermis and
into the dermis may be further defined as a transport phenomenon
and quantified by mass transfer rate(s) and/or fluid mechanics
(e.g., mass flow rate(s)).
[0107] Thus, in some embodiments described herein, the one or more
agents may be delivered to a depth in the epidermis wherein the one
or more agents moves past the basal layer of the viable epidermis
and into the viable dermis. In some aspects described herein, the
one or more agents are then absorbed by one or more susceptible
lymphatic capillary plexus or blood capillaries and then delivered
to one or more susceptible tumors.
[0108] In some embodiments described herein, the one or more agents
may be delivered in a liquid carrier solution. In one aspect, the
tonicity of the liquid carrier may be hypertonic to the fluids
within the blood capillaries or lymphatic capillaries. In another
aspect, the tonicity of a liquid carrier solution may be hypotonic
to the fluids within the blood capillaries or lymphatic
capillaries. In another aspect, the tonicity of a liquid carrier
solution may be isotonic to the fluids within the blood capillaries
or lymphatic capillaries. The liquid carrier solution may further
comprise at least one or more pharmaceutically acceptable
excipients, diluent, cosolvent, particulates, or colloids.
Pharmaceutically acceptable excipients for use in liquid carrier
solutions is known, see, for example, Pharmaceutics: Basic
Principles and Application to Pharmacy Practice (Alekha Dash et al.
eds., 1.sup.st ed. 2013), which is incorporated by reference herein
for its teachings thereof.
[0109] In some embodiments, the one or more agents may then be
directly or indirectly delivered to one or more susceptible tumors
by first delivering the one or more agents to a depth in the skin,
which results in delivery to a susceptible lymphatic capillary
plexus or a blood capillary plexus as described herein. In one
aspect, the targeted delivery of one or more agents to one or more
susceptible tumors comprises delivery to the epidermis, wherein the
one or more agents is absorbed by a susceptible lymphatic capillary
plexus prior to being absorbed by one or more susceptible tumors.
In another aspect, the targeted delivery of one or more agents to
one or more susceptible tumors comprises delivery to the viable
epidermis and/or viable dermis, wherein the one or more agents is
absorbed by a blood capillary plexus prior to being absorbed by one
or more susceptible tumors.
[0110] In some embodiments described herein, the distribution of
depths in the skin, wherein a portion of the one or more agents is
initially delivered, which results in uptake of the one or more
agents by one or more susceptible tumors ranges from about 5 .mu.m
to about 4,500 .mu.m, including each integer within the specified
range. In some aspects, the depth in the skin for initially
delivering one or more agents ranges from about 5 .mu.m to about
2,000 .mu.m, including each integer within the specified range. In
some aspects, the depth in the skin for initially delivering one or
more agents ranges from about 5 .mu.m to about 1,000 .mu.m,
including each integer within the specified range. In some aspects,
the depth in the skin for initially delivering one or more agents
ranges from about 5 .mu.m to about 500 .mu.m, including each
integer within the specified range. In some aspects, the depth in
the skin for initially delivering one or more agents ranges from
about 5 .mu.m to about 250 .mu.m, including each integer within the
specified range. In some aspects, the depth in the skin for
initially delivering one or more agents ranges from about 5 .mu.m
to about 100 .mu.m, including each integer within the specified
range. In some aspects, the average depth in the skin for initially
delivering one or more agents is about 5 .mu.m, about 10 .mu.m,
about 20 .mu.m, about 30 .mu.m, about 40 .mu.m, about 50 .mu.m,
about 60 .mu.m, about 70 .mu.m, about 80 .mu.m, about 90 .mu.m,
about 100 .mu.m, about 125 .mu.m, about 150 .mu.m, about 175 .mu.m,
about 200 .mu.m, about 225 .mu.m, about 250 .mu.m, about 275 .mu.m,
about 300 .mu.m, about 350 .mu.m, about 400 .mu.m, about 450 .mu.m,
about 500 .mu.m, about 550 .mu.m, about 600 .mu.m, about 650 .mu.m,
about 700 .mu.m, about 750 .mu.m, about 800 .mu.m, about 850 .mu.m,
about 900 .mu.m, about 950 .mu.m, about 1,000 .mu.m, about 1,100
.mu.m, about 1,200 .mu.m, about 1,300 .mu.m, about 1,400 .mu.m,
about 1,500 .mu.m, about 1,600 .mu.m, about 1,700 .mu.m, about
1,800 .mu.m, about 1,900 .mu.m, about 2,000 .mu.m, about 2,250
.mu.m, about 2,500 .mu.m, about 2,750 .mu.m, about 3,000 .mu.m,
about 3,250 .mu.m, about 3,500 .mu.m, about 3,750 .mu.m, about
4,000 .mu.m, to about 4,500 .mu.m.
[0111] In some embodiments described herein, one or more agents are
delivered to the viable skin, wherein the distribution of depths in
the viable skin for delivery of the one or more agents is
immediately past the stratum corneum of the epidermis but above the
subcutaneous tissue, which results in uptake of the one or more
agents by one or more susceptible tumors. Whether the agent is
within the epidermis or dermis will depend on the thickness of the
epidermis, for example, more shallow depths of delivery comprising
about I .mu.m to about 250 .mu.m past the stratum corneum would be
expected to be within the viable epidermis. Depths greater than 400
.mu.m, 500 .mu.m, or 700 .mu.m would likely be expected to be
within at least a most superficial portion of the viable dermis
(e.g., the papillary dermis). In some aspects, the depth in the
viable skin for delivering one or more agents ranges from about 1
.mu.m to about 5,000 .mu.m beyond the stratum corneum, but still
within the viable skin above the subcutaneous tissue, including
each integer within the specified range. In some aspects, the depth
in the viable skin for delivering one or more agents ranges from
about I .mu.m to about 3,500 .mu.m beyond the stratum corneum, but
still within the viable skin above the subcutaneous tissue,
including each integer within the specified range. In some aspects,
the depth in the viable skin for delivering one or more agents
ranges from about 1 .mu.m to about 2,000 .mu.m beyond the stratum
corneum, but still within the viable skin above the subcutaneous
tissue, including each integer within the specified range. In some
aspects, the depth in the viable skin for delivering one or more
agents ranges from about I .mu.m to about 1,000 .mu.m beyond the
stratum corneum, but still within the viable skin above the
subcutaneous tissue, including each integer within the specified
range. In some aspects, the depth in the viable skin for delivering
one or more agents ranges from about 1 .mu.m to about 500 .mu.m
beyond the stratum corneum, but still within the viable skin above
the subcutaneous tissue, including each integer within the
specified range. In some aspects, the depth in the viable skin for
delivering one or more agents ranges from about 1 .mu.m to about
250 .mu.m beyond the stratum corneum, but still within the viable
skin above the subcutaneous tissue, including each integer within
the specified range. In some aspects, the depth in the viable skin
for delivering one or more agents ranges from about 1 .mu.m to
about 100 .mu.m beyond the stratum corneum, but still within the
viable skin above the subcutaneous tissue, including each integer
within the specified range. In some aspects, the depth in the
viable skin for delivering one or more agents ranges from about 1
.mu.m to about 50 .mu.m beyond the stratum corneum, but still
within the viable skin above the subcutaneous tissue, including
each integer within the specified range. In some aspects, the
average depth in the viable skin for delivering one or more agents
is about 1 .mu.m, about 5 .mu.m, about 10 .mu.m, about 20 .mu.m,
about 30 .mu.m, about 40 .mu.m, about 50 .mu.m, about 60 .mu.m,
about 70 .mu.m, about 80 .mu.m, about 90 .mu.m, about 100 .mu.m,
about 150 .mu.m, about 250 .mu.m, about 350 .mu.m, about 450 .mu.m,
about 550 .mu.m, about 650 .mu.m, about 750 .mu.m, about 850 .mu.m,
about 950 .mu.m, about 1,000 .mu.m, about 1,100 .mu.m, about 1,200
.mu.m, about 1,300 .mu.m, about 1,400 .mu.m, about 1,500 .mu.m,
about 1,600 .mu.m, about 1,700 .mu.m, about 1,800 .mu.m, about
1,900 .mu.m, about 2,000 .mu.m, about 2,250 .mu.m, about 2,500
.mu.m, about 2,750 .mu.m, about 3,000 .mu.m, about 3,250 .mu.m,
about 3,500 .mu.m, about 3,750 .mu.m, about 4,000 .mu.m, about
4,500 .mu.m, or about 5,000 .mu.m beyond the stratum corneum, but
still within the viable skin above the subcutaneous tissue.
[0112] Non-limiting tests for assessing initial delivery depth in
the skin may be invasive (e.g., a biopsy) or non-invasive (e.g.,
imaging). Conventional non-invasive optical methodologies may be
used to assess delivery depth of an agent into the skin including
remittance spectroscopy, fluorescence spectroscopy, photothermal
spectroscopy, or optical coherence tomography (OCT). Imaging using
methods may be conducted in real-time to assess the initial
delivery depths. Alternatively, invasive skin biopsies may be taken
immediately after administration of an agent, followed by standard
histological and staining methodologies to determine delivery depth
of an agent. For examples of optical imaging methods useful for
determining skin penetration depth of administered agents see
Sennhen, et al., Skin Pharmacol., 6(2), 152-160 (1993), Gotter, et
al., Skin Pharmacol. Physiol., 21, 156-165 (2008), and Mogensen, et
al., Semin. Cutan. Med. Surg., 28, 196-202 (2009), each of which
are incorporated by reference herein for their teachings
thereof.
[0113] In some embodiments described herein are methods for the
extended delivery (or administration) of one or more agents
described herein. In some aspects, the one or more agents is
delivered over a period of time from about 0.5 hours to about 72
hours, including each integer of time within the specified range.
In some aspects, the one or more agents is delivered over a period
of time from about 0.5 hours to about 48 hours, including each
integer of time within the specified range. In some aspects, the
one or more agents is delivered over a period time from about 0.5
hours to about 24 hours, including each integer of time within the
specified range. In some aspects, the one or more agents is
delivered over a period of time from about 0.5 hours to about 12
hours, including each integer of time within the specified range.
In some aspects, the one or more agents is delivered over a period
of time from about 0.5 hours to about 6 hours, including each
integer of time within the specified range. In some aspects, the
one or more agents is delivered over a period of time of about 0.5
hours, about I hours, about 1.5 hours, about 2 hours, about 2.5
hours, about 3 hours, about 3.5 hours, about 4 hours, about 4.5
hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours,
about 9 hours, about 10 hours, about 11 hours, about 12 hours,
about 16 hours, about 20 hours, about 24 hours, about 28 hours,
about 32 hours, about 36 hours, about 40 hours, about 44 hours,
about 48 hours, about 52 hours, about 56 hours, about 60 hours,
about 64 hours, about 68 hours, or about 72 hours.
[0114] In some embodiments described herein, one or more agents in
a liquid carrier solution are administered to an initial
approximate space in the skin. The one or more agents in a liquid
carrier solution initially delivered to the skin (e.g., prior to
any subsequent movement or diffusion) may be distributed within, or
encompassed by an approximate three dimensional volume of the skin.
Thus, as further described herein, the one or more initially
delivered agents exhibits a Gaussian distribution of delivery
depths and will also have a Gaussian distribution within a three
dimensional volume of the skin tissue. In some aspects, the one or
more agents in a liquid carrier solution may be administered to the
skin, wherein the tissue volume comprising the one or more agents
in a liquid carrier solution is about 0.7 mm.sup.3 to about 2,500
mm.sup.3, including each integer within the specified range. In
some aspects, the one or more agents in a liquid carrier solution
may be administered to the skin, wherein the total three
dimensional surface area of the administered liquid carrier
solution comprising the one or more agents is about 18 mm.sup.2 to
about 20,000 mm.sup.2, including each integer within the specified
range. In some aspects, the one or more agents in a liquid carrier
solution may be administered to the skin, wherein the three
dimensional surface area to volume ratio of the administered liquid
carrier solution comprising the one or more agents is about 35
mm.sup.-1 to about 5 mm.sup.-1, including each integer within the
specified range. The exemplified volume, surface area, and surface
area to volume ratios may vary depending on the local physiological
administration site, size of the delivery device, delivery depth,
and disease to be treated.
[0115] The tissue volume, surface area, and surface area to volume
ratio of a delivered agent may be determined by using standard
geometric calculations following measuring the overall dimensions
(width and length) of the delivery device in contact with the skin
of a subject and the deepest delivery depth of an initially
administered agent using the standard methods and techniques of
measuring delivery depth as described herein.
[0116] In some embodiments described herein, multiple dosages of
one or more agents in a liquid carrier solution as described herein
is simultaneously administered to the skin for targeted delivery to
one or more susceptible tumors. In some aspects, one or more agents
in a liquid carrier solution are simultaneously administered in
between 2 and 50,000 sub doses, including each integer within the
specified range. In some aspects, one or more agents in a liquid
carrier solution are simultaneously administered in between 2 and
25,000 sub doses. In some aspects, one or more agents in a liquid
carrier solution are simultaneously administered in between 2 and
15,000 sub doses, including each integer within the specified
range. In some aspects, one or more agents in a liquid carrier
solution are simultaneously administered in between 2 and 10,000
sub doses, including each integer within the specified range. In
some aspects, one or more agents in a liquid carrier solution are
simultaneously administered in between 2 and 5,000 sub doses,
including each integer within the specified range. In some aspects,
one or more agents in a liquid carrier solution are simultaneously
administered in between 2 and 1,000 sub doses, including each
integer within the specified range. In some aspects, one or more
agents in a 10 liquid carrier solution are simultaneously
administered in between 2 and 500 sub doses, including each integer
within the specified range. In some aspects, one or more agents in
a liquid carrier solution are simultaneously administered in
between 2 and 250 sub doses, including each integer within the
specified range. In some aspects, one or more agents in a liquid
carrier solution are simultaneously administered in between 2 and
150 sub doses, including each integer within the specified range.
In some aspects, one or more agents in a liquid carrier solution
are simultaneously administered in between 2 and 100 sub doses. In
some aspects, one or more agents in a liquid carrier solution are
simultaneously administered in between 2 and 50 sub doses,
including each integer within the specified range. In some aspects,
one or more agents in a liquid carrier solution are simultaneously
administered in between 2 and 25 sub doses, including each integer
within the specified range. In some aspects, one or more agents in
a liquid carrier solution are simultaneously administered in
between 2 and 15 sub doses, including each integer within the
specified range. In some aspects, one or more agents in a liquid
carrier solution are simultaneously administered in between 2 and
10 sub doses. In some aspects, one or more agents in a liquid
carrier solution is simultaneously administered in about 2 sub
doses, about 5 sub doses, about 10 sub doses, about 15 sub doses,
about 20 sub doses, about 25 sub doses, about 30 sub doses, about
35 sub doses, about 40 sub doses, about 45 sub doses, about 50 sub
doses, about 75 sub doses, about 80 sub doses, about 85 sub doses,
about 90 sub doses, about 95 sub doses, about 100 sub doses, about
150 sub doses, about 200 sub doses, about 250 sub doses, about 300
sub doses, about 350 sub doses, about 400 sub doses, about 450 sub
doses, about 500 sub doses, about 600 sub doses, about 700 sub
doses, about 800 sub doses, about 900 sub doses, about 1,000 sub
doses, about 2,000 sub doses, about 3,000 sub doses, about 4,000
sub doses, about 5,000 sub doses, about 6,000 sub doses, about
7,000 sub doses, about 8,000 sub doses, about 9,000 sub doses,
about 10,000 sub doses, about 15,000 sub doses, about 20,000 sub
doses, about 25,000 sub doses, about 30,000 sub doses, about 35,000
sub doses, about 40,000 sub doses, about 45,000 sub doses, or about
50,000 sub doses. In some aspects, the above described sub doses
may be administered by a suitable delivery structure as further
described herein.
[0117] In some embodiments described herein, the flow rate of one
or more administered agents to the skin per single delivery
structure as described herein may be about 0.01 .mu.l per hour to
about 500 .mu.l per hour, including each integer within the
specified range for the targeted delivery to one or more
susceptible tumors. In some aspects, the controlled flow rate of
one or more administered agents per single delivery structure as
described herein may be about 0.01 .mu.l per hour to about 250
.mu.l per hour, including each integer within the specified range
In some aspects, the controlled flow rate of one or more
administered agents per single delivery structure as described
herein may be about 0.01 .mu.l per hour to about 150 .mu.l per
hour, including each integer within the specified range. In some
aspects, the controlled flow rate of one or more administered
agents per single delivery structure as described herein may be
about 0.01 .mu.l per hour to about 100 .mu.l per hour, including
each integer with in the specified range. In some aspects, the
controlled flow rate of one or more administered agents per single
delivery structure as described herein may be about 0.01 .mu.l per
hour to about 50 .mu.l per hour, including each integer within the
specified range. In some aspects, the controlled flow rate of one
or more administered agents per single delivery structure as
described herein may be about 0.01 .mu.l per hour to about 25 .mu.l
per hour, including each integer within the specified range. In
some aspects, the controlled flow rate of one or more administered
agents per single delivery structure as described herein may be
about 0.01 .mu.l per hour, about 0.5 .mu.l per hour, about 1 .mu.l
per hour, about 1.5 .mu.l per hour, about 2 .mu.l per hour, about
2.5 .mu.l per hour, about 3 .mu.l per hour, about 3.5 .mu.l per
hour, about 4 .mu.l per hour, about 4.5 .mu.l per hour, about 5
.mu.l per hour, about 10 .mu.l per hour, about 15 .mu.l per hour,
about 20 .mu.l per hour, about 25 .mu.l per hour, about 30 .mu.l
per hour, about 35 .mu.l per hour, about 40 .mu.l per hour, about
45 .mu.l per hour, about 50 .mu.l per hour, about 60 .mu.l per
hour, about 70 .mu.l per hour, about 80 .mu.l per hour, about 90
.mu.l per hour, about 100 .mu.l per hour, about 125 .mu.l per hour,
about 150 .mu.l per hour, about 175 .mu.l per hour, about 200 .mu.l
per hour, about 225 .mu.l per hour, about 250 .mu.l per hour, about
300 .mu.l per hour, about 350 .mu.l per hour, about 400 .mu.l per
hour, about 450 .mu.l per hour, about 500 .mu.l per hour.
[0118] In some embodiments described herein, the overall controlled
flow rate of one or more administered agents to a subject as
described herein may be from about 0.02 .mu.l per hour to about
50,000 .mu.l per hour, including each integer within the specified
range, which results in uptake of the one or more agents by one or
more susceptible tumors. In some aspects, the overall controlled
flow rate of one or more administered agents described herein may
be from about 0.02 .mu.l per hour to about 25,000 .mu.l per hour,
including each integer with in the specified range. In some
aspects, the overall controlled flow rate of one or more
administered agents described herein may be from about 0.02 .mu.l
per hour to about 15,000 .mu.l per hour, including each integer
within the specified range. In some aspects, the overall controlled
flow rate of one or more administered agents described herein may
be from about 0.02 .mu.l per hour to about 10,000 .mu.l per hour,
including each integer within the specified range. In some aspects,
the overall controlled flow rate of one or more administered agents
described herein may be from about 0.02 .mu.l per hour to about
5,000 .mu.l per hour, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may be from about 0.02
.mu.l per hour to about 2,500 .mu.l per hour, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may be from about 0.02 .mu.l per hour to about 1,250 .mu.l
per hour, including each integer within the specified range. In
some aspects, the overall controlled flow rate of one or more
administered agents described herein may be from about 0.02 .mu.l
per hour to about 500 .mu.l per hour, including each integer within
the specified range. In some aspects, the overall controlled flow
rate of one or more administered agents described herein may be
from about 0.02 .mu.l per hour to about 250 .mu.l per hour,
including each integer within the specified range. In some aspects,
the overall controlled flow rate of one or more administered agents
described herein may be from about 0.02 .mu.l per hour to about 125
.mu.l per hour, including each integer within the specified range.
In some aspects, the overall controlled flow rate of one or more
administered agents described herein may be from about 0.02 .mu.l
per hour to about 50 .mu.l per hour, including each integer within
the specified range. In some aspects, the overall controlled flow
rate of one or more administered agents described herein may be
from about 0.02 .mu.l per hour to about 25 .mu.l per hour,
including each integer within the specified range. In some aspects,
the overall controlled flow rate of one or more administered agents
described herein may be from about 0.02 .mu.l per hour to about 10
.mu.l per hour, including each integer within the specified range.
In some aspects, he overall controlled flow rate of one or more
agents described herein may be about 0.02 .mu.l per hour, about 0.5
.mu.l per hour, about 1 .mu.l per hour, about 1.5 .mu.l per hour,
about 2 .mu.l per hour, about 2.5 .mu.l per hour, about 3 .mu.l per
hour, about 3.5 .mu.l per hour, about 4 .mu.l per hour, about 4.5
.mu.l per hour, about 5 .mu.l per hour, about 10 .mu.l per hour,
about 15 .mu.l per hour, about 20 .mu.l per hour, about 25 .mu.l
per hour, about 30 .mu.l per hour, about 35 .mu.l per hour, about
40 .mu.l per hour, about 45 .mu.l per hour, about 50 .mu.l per
hour, about 60 .mu.l per hour, about 70 .mu.l per hour, about 80
.mu.l per hour, about 90 .mu.l per hour, about 100 .mu.l per hour,
about 125 .mu.l per hour, about 150 .mu.l per hour, about 175 .mu.l
per hour, about 200 .mu.l per hour, about 225 .mu.l per hour, about
250 .mu.l per hour, about 300 .mu.l per hour, about 350 .mu.l per
hour, about 400 .mu.l per hour, about 450 .mu.l per hour, about 500
.mu.l per hour, about 550 .mu.l per hour, about 600 .mu.l per hour,
about 650 .mu.l per hour, about 700 .mu.l per hour, about 750 .mu.l
per hour, about 800 .mu.l per hour, about 850 .mu.l per hour, about
900 .mu.l per hour, about 950 .mu.l per hour, about 1,000 .mu.l per
hour, about 1,250 .mu.l per hour, about 1,500 .mu.l per hour, about
1,750 .mu.l per hour, about 2,000 .mu.l per hour, about 2,250 .mu.l
per hour, about 2,500 .mu.l per hour, about 2,750 .mu.l per hour,
about 3,000 .mu.l per hour, about 3,250 .mu.l per hour, about 3,500
.mu.l per hour, about 3,750 .mu.l per hour, about 4,000 .mu.l per
hour, about 4,250 .mu.l per hour, about 4,500 .mu.l per hour, about
4,750 .mu.l per hour, about 5,000 .mu.l per hour, about 5,500 .mu.l
per hour, about 6,000 .mu.l per hour, about 6,500 .mu.l per hour,
about 7,000 .mu.l per hour, about 7,500 .mu.l per hour, about 8,000
.mu.l per hour, about 8,500 .mu.l per hour, about 9,000 .mu.l per
hour, about 9,500 .mu.l per hour, about 10,000 .mu.l per hour,
about 10,000 .mu.l per hour, about 20,000 .mu.l per hour, about
30,000 .mu.l per hour, about 40,000 .mu.l per hour, or about 50,000
.mu.l per hour.
[0119] In some embodiments described herein, the combined overall
controlled flow rate of one or more agents administered to the skin
of a subject as described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 50,000 .mu.l/hr/cm.sup.2, including each
integer within the specified range based on the total surface area
of a delivery device that is in contact with the skin of the
subject as further described herein. In the following aspects the
rate of delivery based on the total surface areas described herein
results in uptake of the one or more agents by one or more
susceptible tumors. In one aspect, the total surface area of a
delivery device refers to the two dimensional surface area of the
delivery device backing substrate that is in contact with the skin
of a subject. In another aspect, the total surface area of a
delivery device refers to the combined total of the two dimensional
cross sectional surface areas of each of the independent delivery
structures that are in contact with the skin of a subject. In some
aspects, the overall controlled flow rate of one or more
administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 50,000 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 15,000
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 10,000 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 5,000
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 2,500 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 1,250
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 500 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 250
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 125 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 50
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 25 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, the overall
controlled flow rate of one or more administered agents described
herein may range from about 0.02 .mu.l/hr/cm.sup.2 to about 10
.mu.l/hr/cm.sup.2, including each integer within the specified
range. In some aspects, the overall controlled flow rate of one or
more administered agents described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 5 .mu.l/hr/cm.sup.2, including each
integer within the specified range. In some aspects, be overall
controlled flow rate of one or more agents described herein may be
about 0.02 .mu.l/hr/cm.sup.2, about 0.5 .mu.l/hr/cm.sup.2, about 1
.mu.l/hr/cm.sup.2, about 1.5 .mu.l/hr/cm.sup.2, about 2
.mu.l/hr/cm.sup.2, about 2.5 .mu.l/hr/cm.sup.2, about 3
.mu.l/hr/cm.sup.2, about 3.5 .mu.l/hr/cm.sup.2, about 4
.mu.l/hr/cm.sup.2, about 4.5 .mu.l/hr/cm.sup.2, about 5
.mu.l/hr/cm.sup.2, about 10 .mu.l/hr/cm.sup.2, about 15
.mu.l/hr/cm.sup.2, about 20 .mu.l/hr/cm.sup.2, about 25
.mu.l/hr/cm.sup.2, about 30 .mu.l/hr/cm.sup.2, about 35
.mu.l/hr/cm.sup.2, about 40 .mu.l/hr/cm.sup.2, about 45
.mu.l/hr/cm.sup.2, about 50 .mu.l/hr/cm.sup.2 about 60
.mu.l/hr/cm.sup.2, about 70 .mu.l/hr/cm.sup.2 about 80
.mu.l/hr/cm.sup.2, about 90 .mu.l/hr/cm.sup.2, about 100
.mu.l/hr/cm.sup.2, about 125 .mu.l/hr/cm.sup.2, about 150
.mu.l/hr/cm.sup.2, about 175 .mu.l/hr/cm.sup.2, about 200
.mu.l/hr/cm.sup.2, about 225 .mu.l/hr/cm.sup.2, about 250
.mu.l/hr/cm.sup.2, about 300 .mu.l/hr/cm.sup.2, about 350
.mu.l/hr/cm.sup.2, about 400 .mu.l/hr/cm.sup.2, about 450
.mu.l/hr/cm.sup.2, about 500 .mu.l/hr/cm.sup.2, about 550
.mu.l/hr/cm.sup.2, about 600 .mu.l/hr/cm.sup.2, about 650
.mu.l/hr/cm.sup.2, about 700 .mu.l/hr/cm.sup.2, about 750
.mu.l/hr/cm.sup.2, about 800 .mu.l/hr/cm.sup.2, about 850
.mu.l/hr/cm.sup.2, about 900 .mu.l/hr/cm.sup.2, about 950
.mu.l/hr/cm.sup.2, about 1,000 .mu.l/hr/cm.sup.2, about 1,250
.mu.l/hr/cm.sup.2, about 1,500 .mu.l/hr/cm.sup.2, about 1,750
.mu.l/hr/cm.sup.2, about 2,000 .mu.l/hr/cm.sup.2, about 2,250
.mu.l/hr/cm.sup.2, about 2,500 .mu.l/hr/cm.sup.2 about 2,750
.mu.l/hr/cm.sup.2, about 3,000 .mu.l/hr/cm.sup.2, about 3,250
.mu.l/hr/cm.sup.2, about 3,500 .mu.l/hr/cm.sup.2, about 3,750
.mu.l/hr/cm.sup.2, about 4,000 .mu.l/hr/cm.sup.2, about 4,250
.mu.l/hr/cm.sup.2, about 4,500 .mu.l/hr/cm.sup.2, about 4,750
.mu.l/hr/cm.sup.2, about 5,000 .mu.l/hr/cm.sup.2, about 5,500
.mu.l/hr/cm.sup.2 about 6,000 .mu.l/hr/cm.sup.2 about 6,500
.mu.l/hr/cm.sup.2, about 7,000 .mu.l/hr/cm.sup.2, about 7,500
.mu.l/hr/cm.sup.2, about 8,000 .mu.l/hr/cm.sup.2, about 8,500
.mu.l/hr/cm.sup.2, about 9,000 .mu.l/hr/cm.sup.2, about 9,500
.mu.l/hr/cm.sup.2, about 10,000 .mu.l/hr/cm.sup.2, about 20,000
.mu.l/hr/cm.sup.2, about 30,000 .mu.l/hr/cm.sup.2, about 40,000
.mu.l/hr/cm.sup.2, or about 50,000 .mu.l/hr/cm.sup.2.
[0120] In some embodiments described herein, the flow rate of one
or more administered agents to the skin per single delivery
structure as described herein may be about 0.01 .mu.l per hour to
about 500 .mu.l per hour for the targeted delivery to one or more
susceptible tumors.
[0121] In some embodiments described herein, the overall controlled
flow rate of one or more administered agents to a subject as
described herein may be from about 0.2 .mu.l per hour to about
50,000 .mu.l per hour, which results in uptake of the one or more
agents by one or more susceptible tumors.
[0122] In some embodiments described herein, the combined overall
controlled flow rate of one or more agents administered to the skin
of a subject as described herein may range from about 0.02
.mu.l/hr/cm.sup.2 to about 50,000 .mu.l/hr/cm.sup.2 based on the
total surface area of a delivery device that is in contact with the
skin of the subject as further described herein. In one aspect, the
total surface area of a delivery device refers to the two
dimensional surface area of the delivery device backing substrate
that is in contact with the skin of a subject. In another aspect,
the total surface area of a delivery device refers to the combined
total of the two dimensional cross sectional surface areas of each
of the independent delivery structures that are in contact with the
skin of a subject
[0123] In some embodiments, the methods described herein provide
for increased delivery of one or more agents to one or more
lymphatic tissues for the targeted delivery to one or more
susceptible tumors. In some aspects, the one or more agents travel
through the lymphatic vasculature to one or more lymph nodes or one
or more sites of a tumor. Without being bound by any theory, it is
thought that the uptake of one or more agents by the lymphatic
vasculature allows for delivery of the one or more agents to a
location surrounding the susceptible tumor. Furthermore, tumors
elicit lymphangiogenic responses, which may further increase the
local lymphatic vasculature peripheral to a susceptible tumor
allowing for drug delivery. Also, without being limited theory, it
is further thought that increasing delivery of one or more
bioactive agents (e.g., an anti-cancer agent) through the lymphatic
vasculature may limit the metastatic dissemination of the tumor
through the peripheral lymphatics by exerting a localized cytotoxic
or therapeutic effect against potential metastatic cancer
cells.
[0124] As further described herein, the physiology and hydrostatics
of the lymphatic vasculature plays an important role in mammalian
physiology and a yet untapped resource for the delivery of agents
to tumors. The lymphatic vasculature comprises all of the lymphatic
endothelial cells making up the lymphatic capillaries, larger
lymphatic vessels, and collecting ducts. The fluid within the
lymphatic vasculature and all bio-materials in this fluid
eventually drain into one or more lymph nodes and ultimately into
the blood stream to enter the systemic circulation. For a complete
review of the lymphatic physiology, see, William N. Charman and
Valentino J. Stella, Lymphatic Transport of Drugs (1992), which is
incorporated by reference herein in its entirety.
[0125] The lymphatic system is a part of the immune system,
protecting the body against infection and invasion by foreign
organisms. Lymphocytes and macrophages patrol most of the body's
tissues for invading viruses, bacteria, tumor cells, foreign
proteins, toxins, damaged and dying cells, and foreign cells,
including, foreign tissue grafts. Lymph vessels communicate with
most tissues, transporting the lymph fluid that carries the immune
cells to the lymph nodes and lymphatic organs, such as the spleen
and thymus. The lymphatic vessels, also referred to as lymphatics
or lymphatic vasculature, are a network of thin opaque tube-like
structures that branch, like blood vessels, into tissues throughout
the body. In mammals, including humans, most tissues and organs are
drained by the lymphatic system.
[0126] Unlike the circulatory system, the lymphatic system is not
closed and has no central pump. The lymphatic system forms a
one-way flow system towards the heart. An elaborate network of
lymph capillaries drains interstitial fluid from the tissues, after
which, this fluid is referred to as lymph. The lymphatics enter all
tissues except epithelia, brain, spinal cord, and bone marrow. A
few connective tissues, such as cartilage and the cornea, have no
blood vessels and also lack lymphatics. The lymph moves slowly and
under low pressure from peristaltic contraction.
[0127] These lymphatic capillaries are ten to fifty micrometers in
diameter. They start from a blind sac, or from anastomosing
vessels. The endothelium is a single layer, with an incomplete
basement membrane. They possess gap junctions that are highly
permeable to plasma proteins and large particles, including, for
example, carbon particles, pathogens, such as viruses, bacterial
cells, and parasites, cells, including, for example, immune cells
and tumor cells, and cellular debris. The lymphatic capillaries
have one-way valves, which ensure flow is only in one direction.
When the pressure of the interstitial fluid outside of the
lymphatic capillary is greater than the pressure inside the
capillary, the flaps open allowing for fluid to enter. Conversely,
when the pressure is greater inside the capillary, the flap is
forced shut, precluding any lymph from leaking out of the vessel.
During inflammation, the capillaries develop further openings that
allow for the uptake of even larger molecules and cellular
debris.
[0128] Lymph flows from capillaries into collecting lymphatics
where it encounters the first of many lymph nodes. These "afferent"
lymphatic vessels bring lymph to a lymph node and the "efferent"
lymphatic vessels take the lymph away from a lymph node. Lymph is a
colorless, watery fluid originating from interstitial fluid. Lymph
originates as blood plasma lost from the capillary beds of the
circulatory system, which leaks out into the surrounding tissues.
Although the capillaries of the circulatory system lose only about
1% of the volume of the fluid that passes through them to the
interstitial tissue; however, so much blood circulates that the
cumulative fluid loss in the average human body is about three
liters per day. The lymphatic system recaptures this fluid by
diffusion into lymph capillaries, and filters it through the
various lymph nodes and returns it to the circulatory system by way
of the thoracic duct. Once within the lymphatic system the fluid is
called lymph, and has almost the same composition as the original
interstitial fluid.
[0129] Lymphatic capillaries are ubiquitous found throughout the
body. Non-limiting examples of such locations include the viable
skin (dermis), tendons, striated muscle, muscle sheaths, the
periosteum of bone, joint capsules, under the mesothelium lining of
pleural, peritoneal, and pericardial cavities, the alimentary
canal, salivary glands, liver, spleen, nasal cavity, trachea,
bronchi, thyroid gland, thymus, adrenal gland, kidney, bladder,
urethra, prostate, testis, uterus, ovary, and heart.
[0130] The lymph nodes filter lymph, with an internal honeycomb of
connective tissue filled with lymphocytes that collect and destroy
bacteria and viruses. Lymph nodes also produce lymphocytes and
antibodies. When the body is fighting an infection, these
lymphocytes multiply rapidly and produce a characteristic swelling
of the lymph nodes. Lymph is transported to progressively larger
lymphatic vessels culminating in the right lymphatic duct (for
lymph from the right upper body) and the thoracic duct (for the
rest of the body). These ducts drain into the circulatory system at
the right and left subclavian veins, near the shoulders. Along the
network of lymphatic vessels are a series of various lymphatic
tissues and organs, including lymphatic nodules, Peyer's patches,
tonsils, lymph nodes, the thymus, and the spleen.
[0131] Lymphatic nodules are transient clusters of lymphocytes that
form at sites of infection and then disappear. No capsule or
external covering separates nodules from the surrounding cells and
fluids, which percolates directly into the nodules. The lymph nodes
encapsulate many lymphatic nodules within a tough capsule and are
supplied with blood vessels and lymphatics. Lymph nodes filter the
lymph delivered to them by lymphatic vessels. Thus, lymph nodes
filter the lymph draining from the lymphatic capillary bed in which
the lymph node is situated. Peyer's patches are larger nodular
clusters of lymphocytes located in the walls of the intestines and
the tonsils are pockets of nodular tissue enfolded into the mucosa
of the pharynx. Peyer's patches and the tonsils are situated to
intercept antigens from the digestive and respiratory tracts,
respectively.
[0132] The spleen, lymph nodes, and accessory lymphoid tissue
(including the tonsils and appendix) are the secondary lymphoid
organs. These organs are made up of a scaffolding of connective
tissue that supports circulating B- and T-lymphocytes and other
immune cells, including, for example, macrophages, dendritic cells,
and eosinophils. When microorganisms invade the body or the body
encounters other antigens, the antigens are typically transported
from the tissue to the lymph. The lymph is carried in the lymph
vessels to regional lymph nodes.
[0133] In the lymph nodes, the macrophages and dendritic cells
phagocytose antigens, process antigens, and present antigens to
lymphocytes, which can then start producing antibodies or serve as
memory cells to recognize the antigens again in the future. Lymph
and lymphoid tissue thus contain antibodies and immune cells.
[0134] There is a broad range of lymphatic absorption rates of
fluid from the interstitial tissues. For example, it has been
estimated that the percentage of water being evacuated from
intestinal location via the lymphatics ranges anywhere from 1% to
nearly 85% with other estimates indicating that the lymphatic
system is responsible for absorbing between 15 and 20% of
interstitial fluids. This discrepancy is likely due to the
immediate physiological state of the lymphatic tissue measured.
[0135] Lymphatic capillaries are distributed widely throughout the
skin in mammals. Particularly, certain areas such as the fingers
and palms and plantar surfaces of the feet and toes and the scrotum
have been found to have the highest distribution of lymphatics. The
skin lymphatics consist principally of superficial lymphatic plexus
in the dermis extending upwards to the outer two thirds portions of
the dermal structure into the papillary dermis. Deeper plexus lie
within the dermis near the subcutaneous tissue boundary in areas of
the reticular dermis. In general, very little or no lymphatic
tissue is found within the epidermis or subcutaneous tissue layers.
The lymphatics are typically more uniform in areas of the skin that
have thicker dermal layers (e.g., the palmar surface of the hands
and plantar surface of the feet). Similar to the intestinal
tissues, absorption of interstitial fluid and proteins in the skin
is highly variable, for example, in animal models the lymph flow in
skin areas is approximately I ml/hr/100 g of tissue, which may
increase by over 10 fold depending on the local physiology
surrounding the lymphatic vasculature. Various factors have been
found to affect lymphatic absorption including venous pressure,
contraction of surrounding tissues and blood vessels, and
respiration rates. For example, Starling's equation describes the
generation of interstitial fluid by the competition of hydrostatic
and oncotic forces across semipermeable capillary walls. Thus,
increased hydrostatic pressure or reduced oncotic pressure within a
blood vessel, or increased capillary permeability, will tend to
promote interstitial fluid volume and subsequent fluid absorption
by the lymphatic capillaries or result in oedema.
[0136] The absorption of proteins and lipids is likewise highly
varied and depends largely on fluid absorption rates, location
within the skin, and molecular size. In general, the size and lipo
or hydrophilicity of a molecule plays a large role in its relative
absorption. For example, and without being limited by any theory,
it is thought that molecules smaller than 10 kDa are absorbed by
the blood capillaries and lymphatic capillaries at approximately
the same rate, whereas molecules larger than 20 kDa may more likely
enter into the lymphatics, depending on the physiological status of
a given local lymphatic capillary as described above.
[0137] Thus, it has been widely appreciated that delivery to the
lymphatic system would be highly desirable due to the ubiquitous
nature of the lymphatic capillaries and the capability to absorb a
plethora of differently sized agents. The above mentioned
involvement of the lymphatic system in inflammation and the
occurrence and dissemination of various cancers lend an important
alternative route for both the local and systemic treatment of
cancer and cancer tumors.
[0138] In some embodiments described herein, one or more agents are
directly delivered to a position within the epidermis. In some
aspects, the one or more agents diffuse, move, flow, or migrate to
a position in proximity to the lymphatic vasculature. As described
herein, this placement within the epidermis following the methods
described herein results in the diffusion or movement of an agent
through the epidermis and the viable epidermis and into the top
layers of the dermis. This type of movement provided by the
delivery methods described herein allows for direct contact of an
agent to the most superficially present lymphatic capillary bed(s)
or otherwise known as a lymphatic drainage bed or lymphatic
capillary plexus, which physiologically functions to drain
interstitial fluid for a given location to the rest of the
lymphatic system. The localized delivery of one or more agents to a
lymphatic capillary bed may result in the delivery of the agent to
the first lymph nodes draining the lymphatic capillary bed, also
referred to as "primary" lymph nodes. In some aspects, the
localized delivery of one or more agents may also result in the
delivery of the agent to additional lymph nodes downstream of the
primary lymph nodes, also referred to as "secondary" lymph nodes.
In some aspects, the agent may eventually enter the blood stream
and be delivered systemically to one or more tumors. In some
aspects, the agent may be delivered through the lymphatic
vasculature inside of or in close proximity of a solid tumor or
delivered to a tumor present within one or more lymph nodes.
[0139] Therefore, some embodiments described herein include methods
for the localized delivery of one or more agents to a lymphatic
tissue comprising one or more lymphatic capillaries, lymphatic
nodules, lymph nodes, Peyer's patches, and/or the tonsils. In some
aspects, the methods described herein are suitable for delivery to
one or more lymph nodes in any tissue or region of the body.
Suitable non-limiting examples comprise lymph nodes found in the
hands, the feet, thighs (femoral lymph nodes), arms, legs, underarm
(the axillary lymph nodes), the groin (the inguinal lymph nodes),
the neck (the cervical lymph nodes), the chest (pectoral lymph
nodes), the abdomen (the iliac lymph nodes), the popliteal lymph
nodes, parasternal lymph nodes, lateral aortic lymph nodes,
paraaortic lymph nodes, submental lymph nodes, parotid lymph nodes,
submandibular lymph nodes, supraclavicular lymph nodes, intercostal
lymph nodes, diaphragmatic lymph nodes, pancreatic lymph nodes,
cisterna chyli, lumbar lymph nodes, sacral lymph nodes, obturator
lymph nodes, mesenteric lymph nodes, mesocolic lymph nodes,
mediastinal lymph nodes, gastric lymph nodes, hepatic lymph nodes,
and splenic lymph nodes.
[0140] In some embodiments described herein are methods for
delivering one or more agents to a susceptible lymphatic capillary
plexus of a mammal for the targeted delivery of one or more agents
to one or more susceptible tumors. In some aspects, the one or more
agents are delivered to a tissue area having a susceptible
lymphatic capillary plexus. In some aspects, the one or more agents
are delivered to a viable area of the skin having a susceptible
lymphatic capillary plexus. In some aspects, the susceptible
lymphatic capillary plexus is located within a region of the viable
dermis as described herein. In some aspects, the susceptible
lymphatic capillary plexus readily absorbs one or more agents from
the local surrounding interstitial tissue. In some aspects, the
susceptible lymphatic capillary plexus is susceptible to absorbing
one or more agents from the local surrounding interstitial tissue
due to the local physiological environment. In some aspects, the
local physiological environment is susceptible to absorbing one or
more agents due to the presence of inflammation, higher
interstitial fluid pressure compared to intralymphatic pressure,
higher blood capillary fluid exchange rates, tissue contraction,
respiration or a combination of factors thereof. In one aspect, the
susceptible lymphatic capillary plexus has a lower pressure inside
the lymphatic capillary compared to the surrounding interstitial
fluid, wherein the one or more agents is located, thereby
increasing the local absorption rate of the one or more agents by
the lymphatic capillary plexus. In another aspect, the susceptible
lymphatic capillary plexus and the one or more agents are located
within an area of inflammation (e.g., stemming from an incidence of
cancer), wherein local inflammation promotes the porosity of the
lymphatic capillary plexus, thereby increasing the local absorption
rate of one or more agents delivered thereto.
[0141] In some embodiments described herein are methods for
delivering one or more agents to an area within the viable skin
(e.g., the viable dermis) having one or more susceptible lymphatic
capillary plexus. In some aspects, the delivery method comprises
administering one or more agents in a suitable vehicle to the skin
at a controlled rate, wherein the one or more agents surrounds one
or more susceptible lymphatic capillary plexus. In some aspects,
the one or more agents are delivered as a suspension or solution in
a liquid carrier. Without being bound any theory, it is thought
that administering one or more agents in a liquid carrier solution
at a controlled rate, wherein the one or more lymphatic capillary
plexus are surrounded by the solution containing the one or more
agents may enhance its absorption by the lymphatic capillary
plexus. Again, without being bound by any theory, it is thought
that the absorption of solution containing the one or more agents
may be due to increases in the local dermal interstitial fluid
pressure around the lymphatic capillary plexus.
[0142] In some embodiments described herein, are methods for
matching the physiological absorption rate of one or more
susceptible lymphatic capillary plexus with a liquid carrier
solution having one or more agents. In some aspects described
herein, the methods for matching the tissue absorption rate of a
susceptible lymphatic capillary plexus comprises increasing the
local tissue fluid pressure or tissue hydrostatic pressure
surrounding one or more lymphatic capillary plexus. The absorption
rate of a lymphatic capillary plexus will depend upon numerous
factors as further described herein (e.g., vascular capillary flow
rates and filtration rates, tissue oncotic pressure and hydrostatic
pressure, and tissue compliance, etc.). Without being bound by any
theory, generally a relatively small increase in local interstitial
tissue hydrostatic pressure increases the rate of lymphatic
absorption. At lower tissue interstitial tissue absorption rates,
the lymphatics are the primary route of fluid removal. Various
proteins and other bioparticles are carried with the interstitial
fluid as it drains from the interstitial tissues fluids into the
lymphatic capillaries. The resulting increases in tissue
hydrostatic pressure without a significant lowering of the tissue
oncotic pressure preferentially forces fluid into the lymphatics
and not the blood capillaries due to the general increased
hydraulic conductance of the lymphatic vessels. In contrast, at
relatively higher interstitial tissue absorption rates, the
capillaries are generally the principal route of fluid removal.
This is typically because as increased fluid is pushed into the
interstitial tissues, the tissue oncotic pressure decreases forcing
fluid into the blood capillaries due to the relatively high blood
capillary oncotic pressure. Furthermore, the presence of
intermediate to high levels of interstitial tissue fluid increases
pressure on the extracellular matricis increasing tissue compliance
(e.g., tissue expansion) and general lowering of hydrostatic
pressure. This results in decreased lymphatic drainage. Thus,
interstitial hydrostatic and oncotic pressure exert forces on
capillary walls, whereas only tissue pressure has an impact on
lymphatic draining.
[0143] In some embodiments described herein, the administration of
a fluid containing one or more agents in a liquid carrier solution
achieves an increase in the local dermal interstitial fluid
pressure to promote lymphatic uptake of one or more agents. Without
being limited by any theory, it is thought that interstitial fluid
pressures greater than about 1 mmHg to about 3 mmHg results in
interstitial fluid (e.g., dermal interstitial fluid) lymphatic
draining. This amount of pressure results in the opening of the
pressure responsive lymphatic valves as described herein allowing
for interstitial fluid draining into the lymphatic capillaries. In
some aspects, the area of the dermis, in which there is an
increased dermal interstitial fluid pressure due to the
administration of one or more agents described herein is below the
administration site within the epidermis. In some aspects, the
specific interstitial tissue pressure values maybe from about 1
mmHg to about 15 mmHg. In some aspects, the interstitial tissue
pressure values maybe increased by the methods described herein to
be from about I mmHg to about 10 mmHg. In some aspects, the
specific interstitial tissue pressure values maybe increased by the
methods described herein to be from about 1 mmHg to about 5 mmHg.
In some aspects, the specific interstitial tissue pressure values
maybe increased by the methods described herein to be greater than
about 1 mmHg, greater than about 2 mmHg, greater than about 3 mmHg,
greater than about 4 mmHg, greater than about 5 mmHg, greater than
about 6 mmHg, greater than about 7 mmHg, greater than about 8 mmHg,
greater than about 9 mmHg, greater than about 10 mmHg, greater than
about 11 mmHg, greater than about 12 mmHg, greater than about 13
mmHg, greater than about 14 mmHg, greater than about 15 mmHg,
greater than about 16 mmHg, greater than about 17 mmHg, greater
than about 18 mmHg, greater than about 19 mmHg, or greater than
about 20 mm Hg.
[0144] Any method for assessing interstitial fluid pressure known
in the art may be used. For example, the micropipette, the
wick-in-needle, or wick catheter techniques have shown high levels
of intra assay precision and may be used to assess hydrostatic
pressure surrounding a section of a probe inserted in to the
interstitial space of a subject (e.g., the dermal interstitial
space) while administering an agent or following the administration
of an agent using the methods described herein. See, for example,
Wiig and Swartz., Phsiol. Rev., 1005-1060, (2012), which is
incorporated by reference herein for its teachings thereof.
[0145] In some embodiments described herein, the methods for
controlled delivery described herein result in one or more active
drug substances being deposited in one or more lymph nodes or
lymphatic tissues. In some aspects, the concentration of the one or
more active drug substances is about 0.5% to about 75% of the
initial dosage per gram of lymph node tissue, including each
integer within the specified range. In some aspects, the
concentration of the one or more active drug substances is about
0.5% to about 50% of the initial dosage per gram of lymph node
tissue, including each integer within the specified range. In some
aspects, the concentration of the one or more active drug
substances is about 0.5% to about 25% of the initial dosage per
gram of lymph node tissue, including each integer within the
specified range. In some aspects, the concentration of the one or
more active drug substances is about 0.5% to about 15% of the
initial dosage per gram of lymph node tissue, including each
integer within the specified range. In some aspects, the
concentration of the one or more active drug substances is about
0.5% to about 10% of the initial dosage per gram of lymph node
tissue, including each integer within the specified range. In some
aspects, the concentration of the one or more active drug
substances is about 0.5% to about 5% of the initial dosage per gram
of lymph node tissue, including each integer within the specified
range. In some aspects, the concentration of the one or more active
drug substances is about 10% to about 60% of the initial dosage per
gram of lymph node tissue, including each integer within the
specified range. In some aspects, the concentration of the one or
more active drug substances is about 30% to about 55% of the
initial dosage per gram of lymph node tissue, including each
integer within the specified range. In some aspects, the
concentration of the one or more active drug substances is about
40% to about 50% of the initial dosage per gram of lymph node
tissue, including each integer within the specified range. In some
aspects, the concentration of the one or more active drug
substances is about 0.5%, about 3%, about 4%, about 5%, about 6%,
about 7%, about 8%, about 9%, about 10%, about 15%, about 20%,
about 25%, about 30%, about 35%, about 40%, about 45%, about 50%,
about 55%, about 60%, about 65%, about 70%, or about 75% of the
initial dosage per gram of lymph node tissue.
[0146] In some embodiments described herein, the methods for
controlled delivery described herein result in one or more active
drug substances being deposited in one or more lymph nodes, wherein
the ratio of the initial dose of one or more agents localized per
gram of lymph node tissue to whole blood tissue is from about 2:1
to about 50:1 after about 36 hours, including all ratios within the
specified range. In some aspects, the ratio of the initial dose of
one or more agents localized per gram of lymph node tissue to whole
blood tissue is from about 2:1 to about 25:1 after about 36 hours,
including all ratios within the specified range. In some aspects,
the ratio of the initial dose of one or more agents localized per
gram of lymph node tissue to whole blood tissue is from about 2:1
to about 15:1 after about 36 hours, including all ratios within the
specified range. In some aspects, the ratio of the initial dose of
one or more agents localized per gram of lymph node tissue to whole
blood tissue is from about 2:1 to about 10:1 after about 36 hours,
including all ratios within the specified range. In some aspects,
the ratio of the initial dose of one or more agents localized per
gram of lymph node tissue to whole blood tissue is from about 2:1
to about 5:1 after about 36 hours, including all ratios within the
specified range. In some aspects, the ratio of the initial dose of
one or more agents localized per gram of lymph node tissue to whole
blood tissue is about 2:1, about 3:1, about 4:1, about 5:1, about
6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 12:1, about
14:1, about 16:1, about 18:1, about 20:1, about 25:1, about 30:1,
about 35:1, about 40:1, about 45:1, or about 50:1.
[0147] In some embodiments described herein, the methods for
controlled delivery described herein result in one or more active
drug substances being deposited in one or more lymph nodes, wherein
the ratio of the initial dose of one or more agents localized per
gram of lymph node tissue to the skin is from about 0.1:1 to about
3:1 after about 36 hours, including all ratios within the specified
range. In some aspects, the ratio of the initial dose of one or
more agents localized per gram of lymph node tissue to the skin is
from about 0.25:1 to about 3:1 after about 36 hours, including all
ratios within the specified range. In some aspects, the ratio of
the initial dose of one or more agents localized per gram of lymph
node tissue to the skin is from about 0.5:1 to about 3:1 after
about 36 hours, including all ratios within the specified range. In
some aspects, the ratio of the initial dose of one or more agents
localized per gram of lymph node tissue to the skin is from about
1:1 to about 3:1 after about 36 hours, including all ratios within
the specified range. In some aspects, the ratio of the initial dose
of one or more agents localized per gram of lymph node tissue to
the skin is about 0.1:1, about 0.2:1, about 0.4:1, about 0.6:1,
about 0.8:1, or about 1:1, about 2:1, or about 3:1.
[0148] Non-invasive quantification methods for drug biodistribution
and absorption pharmacokinetics by tissues are well known and are
used for assessing the percentage of absorbed drug per initial dose
per gram of tissue. The percent initial dosage of one or more
agents delivered per gram of lymph node tissue as described herein
may be quantified by directly labelling the one or more agents with
a detectable radio label followed by administration of the agent
using the methods described herein. The imaging and quantification
of the radio labelled agent may be assessed using standard positron
emission tomography (PET) or single-photon emission computed
tomography (SPECT), or a combination of these techniques with X-Ray
computed tomography (CT) or magnetic resonance imaging (MRI) see,
for example, Ding and Wu., Theranostics, 2(11), 1040-1053 (2012),
which is incorporated by reference herein for its teachings
thereof. Useful radiolabels may comprise short or long lived
isotopes, such as .sup.11C, .sup.15O, .sup.18F, .sup.68Ga,
.sup.64Cu, .sup.76Br, .sup.89Zr, .sup.124I. The selected radiolabel
will depend on the agent being tested and specific labelling
protocols well known in the art. The percent absorbed initial
dose/gram of lymph node tissue measured initially using either PET
or SPECT imaging may be calculated using standard
radiopbarmaceutical dosimetry and tissue density tables, see,
Bolch, et al., J. Nucl. Med., 50(3), 477 (2009), which is
incorporated by reference herein for its teachings thereof.
[0149] Other comparative methodologies may be utilized to assess
the amount of drug per gram of tissue delivered to a lymphatic
tissue. Suitable comparative methodologies comprise radiolabelling
one or more agents with any of the above described short or long
lived isotopes above and administering the labelled one or more
agents using the methods described herein to a suitable comparative
test subject. The subject may comprise a laboratory animal such as
a rat, guinea pig, mouse, or monkey. To determine the
biodistribution and percent of an initial dose delivered per gram
of lymphatic tissue, lymphatic tissue (e.g., one or more lymph
nodes) amongst other relative organs may be harvested from the
subject animal and the specific radioactivity counts within that
tissue may be measured and quantified using standard well known
techniques and compared to the radioactivity measurements of the
initial dosage.
[0150] In some embodiments described herein, the methods for
controlled delivery described herein result in more of the initial
dosage of one or more agents being absorbed by one or more
susceptible lymphatic capillary plexus compared to other
traditional delivery routes, such as intravenous (i.v.),
subcutaneous (s.c.), intramuscular (i.m.), or intradermal (i.d.)
injection routes or traditional transdermal patches. In some
aspects, the controlled delivery methods described herein result in
approximately a 1.25 fold to about 50 fold increases in the
lymphatic delivery of one or more agents compared to i.v., s.c.,
i.m., or i.d. parenteral delivery routes, including each integer
within the specified range. In some aspects, the controlled
delivery methods described herein result in approximately a 1.25
fold to about 20 fold increase in the lymphatic delivery of one or
more agents compared to i.v., s.c., i.m., or i.d. parenteral
delivery routes, including each integer within the specified range.
In some aspects, the controlled delivery methods described herein
result in approximately a 1.25 fold to about 10 fold increase in
the lymphatic delivery of one or more agents compared to i.v.,
s.c., i.m., or i.d. parenteral delivery routes, including each
integer within the specified range. In some aspects, the controlled
delivery methods described herein result in approximately a 1.25
fold to about 5 fold increase in the lymphatic delivery of one or
more agents compared to i.v., s.c., i.m., or i.d. parenteral
delivery routes, including each integer within the specified
range.
[0151] Assessing the uptake and comparison of an agent delivered
using the methods described herein by one or more lymphatic
capillary plexus may be determined by using one or more imaging
agents attached to an agent or bioactive agent being delivered
using the methods described herein. These imaging agents can be
used to image the lymphatic capillaries and tissues, for example,
one or more lymphatic capillary plexus or lymph node tissues.
Suitable imaging agents may be any agent that is bio-compatible and
has no biological activity or side effects. Exemplary and
non-limiting imaging agents may be one or more agents used for
direct or indirect X-ray lymphangiography imaging, one or more
contrast agents used for magnetic resonance imaging (MRI), one or
more fluorescent imaging agents for fluorescence
microlymphangiography (FML), or one or more fluorescent imaging
agent excitable by tissue-penetrating near-infrared light (NIR) for
use in CG lymphography. The techniques and agents used for
lymphatic imaging are known in the art, see, for example,
Sevick-Muraca, et al., J. Clin Invest., 124(3), 905-914 (2014),
which is incorporated by reference herein for its teachings
thereof. Standard image analysis algorithms and software may be
used to calculate fluorophore intensity of a delivered agent that
is labelled or tagged with an imaging agent as described above and
compared to traditional i.v., s.c., i.m., or i.d. parenteral
delivery routes.
[0152] In some embodiments, the methods for controlled delivery
described herein result in an equivalent blood serum absorption
rate of one or more agents described herein compared to i.v., s.c.,
i.m., or i.d. parenteral delivery routes, while retaining
relatively higher rates of lymphatic delivery as described herein.
Without being bound by any theory, the rate of delivery may be due
to the lymphatic circulation of one or more agents through the
thoracic duct and into the blood circulation. Standard highly
accurate and precise methodologies for measuring blood serum
concentration and therapeutic monitoring at desired time points may
be used that are well known in the art, such as, but not limited
to, radioimmunoassays, high-performance liquid chromatography
(HPLC), fluorescence polarization immunoassay (FPIA), enzyme
immunoassay (EMIT) or enzyme-linked immunosorbent assays (ELISA).
For calculating the absorption rate using the methods described
above, the drug concentration at several time points should be
measured starting immediately following administration and
incrementally thereafter until a C.sub.max value can be established
and the associated absorption rate calculated.
[0153] In some embodiments described herein are methods for the
controlled delivery of one or more agents in a liquid carrier
solution as described herein to the skin for the targeted delivery
of one or more agents to one or more susceptible tumors. In some
aspects, the methods comprise penetrating at least a most
superficial layer of epidermis with a delivery structure described
herein, contacting the epidermis with one or more permeability
enhancers, and administering one or more agents in a liquid carrier
solution in between about 2 and 50,000 sub doses, wherein the sub
doses are administered to the skin (e.g., non-viable epidermis
and/or viable epidermis and/or viable dermis) at a depth of about
10 .mu.m to about 4,500 .mu.m or about 1 .mu.m to about 4,000 .mu.m
beyond a most superficial layer of the epidermis, but still within
the viable skin above the subcutaneous tissue; and wherein the
administration comprises one or more of (a) an administration flow
rate that matches the tissue lymphatic drainage rate; (b) an
overall administration flow rate of about 0.02 .mu.l/hr/cm.sup.2 to
about 50,000 .mu.l/hr/cm.sup.2 based on the surface area of the
delivery device or delivery structures; (c) an interstitial fluid
pressure greater than about 2 mmHg in the local vicinity of one or
more susceptible lymphatic capillary plexus; and (d) delivering the
one or more agents within a liquid carrier to the skin, wherein the
delivered fluid is in contact with or encompassed by a three
dimensional tissue volume of about 0.7 mm.sup.3 to about 2,500
mm.sup.3.
[0154] In some embodiments described herein are methods for the
controlled delivery of one or more active drug substances to the
skin. In some aspects, an overall dose of one or more active drug
substances in a liquid carrier is delivered to one or more
susceptible tumors. In some aspects, an overall dose of one or more
active drug substances in a liquid carrier is delivered first to
one or more susceptible lymphatic capillary plexus followed by a
targeted delivery to one or more susceptible tumors. This overall
dose may comprise between 2 and 50,000 sub doses as described
herein. In some aspects, the overall dose of one or more active
drug substances may comprise about 0.0001 mg/kg of body weight to
about 100 mg/kg body weight, including each integer within the
specified range. In some aspects, the overall dose of one or more
active drug substances may comprise about 0.001 mg/kg of body
weight to about 100 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.01 mg/kg of
body weight to about 100 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.1 mg/kg of
body weight to about 100 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.1 mg/kg of
body weight to about 50 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.1 mg/kg of
body weight to about 25 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.1 mg/kg of
body weight to about 10 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.1 mg/kg of
body weight to about 5 mg/kg body weight, including each integer
within the specified range. In some aspects, the overall dose of
one or more active drug substances may comprise about 0.0001 mg/kg,
about 0.001 mg/kg, about 0.01 mg/kg, about 0.1 mg/kg, about 0.5
mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg,
about 5 mg/kg, about 6 mg/kg, about 7 mg/kg, about 8 mg/kg, about 9
mg/kg, about 10 mg/kg, about 20 mg/kg, about 30 mg/kg, about 40
mg/kg, about 50 mg/kg, about 60 mg/kg, about 70 mg/kg, about 80
mg/kg, about 90 mg/kg, or about 100 mg/kg.
[0155] In some embodiments described herein are methods for the
controlled delivery of one or more active drug substances to the
skin. In some aspects, an overall dose of one or more active drug
substances in a liquid carrier is delivered to one or more
susceptible tumors. In some embodiments, the methods for controlled
delivery described herein result in an equivalent blood serum
absorption rate of one or more agents described herein compared to
i.v., s.c., i.m., or i.d. parenteral delivery routes, while
retaining relatively higher rates of lymphatic delivery as
described herein. Without being bound by any theory, the rate of
delivery may be due to the lymphatic circulation of one or more
agents through the thoracic duct and into the blood
circulation.
[0156] In some embodiments, the methods for controlled delivery
described herein result in physiologically prolonged levels of a
bioactive agent (e.g., an active drug substance) above a known
efficacious therapeutic threshold. In some aspects, the methods of
controlled delivery described herein reduce or prevent the bolus
administration of one or more active drug substances described
herein. In some aspects, this may result in an increased safety
profile of one or more bioactive agents by limiting potentially
dangerous spikes in the systemic plasma circulation of one or more
bioactive agents. Furthermore, the controlled delivery methods
described herein may further increase the therapeutic ratio of one
or more administered active drug substances by lowering the dose
needed for a therapeutic or beneficial effect.
[0157] In some embodiments, the controlled delivery methods
described herein result in one or more agents being retained at the
site of the disease (e.g., within one or more lymph nodes), and not
spread systemically resulting in a reduced occurrence of known side
effects of the active drug substance. For example, diseases
involving inflammation (e.g., cancer, infection, arthritis), one or
more agents may be absorbed by a susceptible lymphatic capillary
plexus and distributed to the sites of inflammation and not further
distribute into the systemic circulation. The one or more agents
may be distributed systemically by circulating through the
lymphatic vasculature and lymphatic tissues through the thoracic
duct and into the systemic blood circulation. Alternatively, the
one or more agents may be distributed systemically by being
directly absorbed by one or more susceptible blood capillary
plexus. In some aspects, the one or more agents may then be
absorbed by one or more susceptible tumors in a subject by
extravasating from the tumor blood vasculature and into the tumor
stroma.
[0158] In some embodiments, the methods for controlled delivery
described herein result in relatively short time to maximal
therapeutic efficacy or T.sub.max, while retaining physiologically
prolonged levels of one or more active drug substances above the
efficacious therapeutic threshold. The T.sub.max may be independent
of the C.sub.max or total blood serum concentration of an active
drug substance and can be assessed by the perception of
amelioration of a disease or condition.
[0159] In some embodiments described herein, the one or more
bioactive agents delivered to the skin and subsequently one or more
susceptible tumors by the methods described herein may comprise an
active drug substance. For example, the active drug substance may
be a compound (e.g., a small molecule), that is capable of acting
on a cellular receptor or surface protein and function as an
agonist, antagonist, inverse agonist, etc., which results in the
modification of a disease pathway and an often efficacious and
beneficial outcome for a subject afflicted with a disease or
disorder as described herein. The active drug substance may exhibit
toxicity to cancer cells or have bactericidal or anti-viral
activity.
[0160] Suitable active drug substances will depend on the disease
or disorder being treated and the tolerance of the subject for
receiving a particular active drug substance. Suitable active drug
substances described herein may be administered regardless of
whether the active drug substance is hydrophilic, lipophilic, or
amphipathic. Active drug substances may be poorly or highly soluble
in an aqueous environment or demonstrate low or high systemic
permeability (e.g., any BCS Class I, II, III, or IV drug).
Furthermore active drug substances described herein may also
comprise any protein drug, such as an antibody (e.g., a humanized
antibody).
[0161] Exemplary active drug substances may comprises a small
molecule. In some aspects, the small molecule may have a molecular
weight of about 50 g/mol to about 1,000 g/mol (i.e., 50 Da-1,000
Da), including each integer within the specified range. In some
aspects, the small molecule may have a molecular weight of about 50
g/mol, about 100 g/mol, about 150 g/mol, about 200 g/mol, about 250
g/mol, about 300 g/mol, about 350 g/mol, about 400 g/mol, about 450
g/mol, about 500 g/mol, about 550 g/mol, about 600 g/mol, about 650
g/mol, about 700 g/mol, about 750 g/mol, about 800 g/mol, about 850
g/mol, about 900 g/mol, about 950 g/mol, or about 1000 g/mol.
[0162] Other suitable active drug substances may comprise a larger
compound or protein. In some aspects, the compound or protein may
have an atomic mass of about I kDa to about 250 kDa, including each
integer within the specified range. In some aspects, the compound
or protein may have an atomic mass of about 1 kDa, about 5 kDa,
about 10 kDa, about 15 kDa, about 20 kDa, about 25 kDa, about 50
kDa, about 75 kDa, about 100 kDa, about 125 kDa, about 150 kDa,
about 175 kDa, about 200 kDa, about 225 kDa, or about 250 kDa.
[0163] In some embodiments described herein are methods for
administering one or more bioactive agents to an animal, preferably
a mammal, and most preferably a human, for preventing, treating, or
ameliorating one or more symptoms associated with a disease,
disorder, or infection, by delivering the one or more bioactive
agents to the skin of subject's skin. The methods described herein
are useful for the treatment or prevention of a disease or disorder
of the lymphatic system, primary or metastatic neoplastic disease
(i.e., cancer). The bioactive agent's may be provided in
pharmaceutically acceptable compositions or formulations as known
in the art or as described herein.
[0164] In some embodiments described herein, the one or more
bioactive agents are present in a liquid carrier as a substantially
dissolved solution, a suspension, or a colloidal suspension. Any
suitable liquid carrier solution may be utilized that meets at
least the United States Pharmacopeia (USP) specifications, and the
tonicity of such solutions may be modified as is known, see, for
example, Remington: The Science and Practice of Pharmacy (Lloyd V.
Allen Jr. ed., 22.sup.nd ed. 2012. Exemplary non-limiting liquid
carrier solutions may be aqueous, semi-aqueous, or non-aqueous
depending on the bioactive agent(s) being administered. For
example, an aqueous liquid carrier may comprise water and any one
of or a combination of a water-miscible vehicles ethyl alcohol,
liquid (low molecular weight) polyethylene glycol, and the like.
Non aqueous carriers may comprise a fixed oil, such as corn oil,
cottonseed oil, peanut oil, or sesame oil, and the like. Suitable
liquid carrier solutions may further comprise any one of a
preservative, antioxidant, complexation enhancing agent, a
buffering agent, an acidifying agent, saline, an electrolyte, a
viscosity enhancing agent, a viscosity reducing agent, an
alkalizing agent, an antimicrobial agent, an antifungal agent, a
solubility enhancing agent or a combination thereof.
[0165] Some embodiments, described herein, include methods of
treating, preventing, reducing the likelihood of, ameliorating, or
managing a disease or disorder in a subject in need thereof, the
method comprising administering to the subject a therapeutically
effective dose or prophylactically effective dose of one or more
bioactive agents (e.g., an active drug substance) to the skin of a
subject in need thereof. In some aspects described herein are
methods of treating, preventing, reducing the likelihood of,
ameliorating, or managing cancer (e.g., treating one or more
susceptible tumors or metastatic diseases thereof) in a subject,
the method comprising administering to the subject a
therapeutically effective dose or prophylactically effective dose
of one or more bioactive agents (e.g., an active drug substance) to
the skin of a subject in need thereof. In some aspects, the methods
of treating or preventing a disease in a subject by delivering one
or more bioactive agents to the skin of a subject is more effective
than conventional delivery routes, e.g., i.v., s.c., i.m., or i.d.
injections.
[0166] In some embodiments described herein, the methods for
controlled delivery described herein result in one or more active
drug substances being deposited in one or more susceptible tumors.
In some aspects, the concentration of the one or more active drug
substances delivered to one or more susceptible tumors is about
0.5% to about 75% of the initial dosage, including each integer
within the specified range. Assessing the percent of a delivered
agent to one or more tumors may be assessed by non-invasive
techniques such as PET or SPECT or a combination of these
techniques with XCT or MRJ as described herein. The percent initial
dosage of the one or more agents delivered to one or more tumors
herein may be quantified by directly labelling the one or more
agents with a detectable radio label followed by administration of
the agent using the methods described herein. The imaging and
quantification of the radio labelled agent may be assessed using
standard positron emission tomography (PET) or single-photon
emission computed tomography (SPECT), or a combination of these
techniques with X-Ray computed tomography (CT) or magnetic
resonance imaging (MRI). See, for example, Ding and Wu.,
Theranostics, 2(11), 1040-20, 1053 (2012), which is incorporated by
reference herein for its teachings thereof. Useful radiolabels may
comprise short or long lived isotopes, such as, but not limited to,
.sup.11C, .sup.15O, .sup.18F, .sup.68Ga, .sup.64Cu, .sup.76Br,
.sup.89Zr, and .sup.124I. The selected radiolabel will depend on
the agent being tested and specific labelling protocols that are
well known in the art. The percent absorbed of the initial dose
delivered to a tumor tissue measured initially using either PET or
a combination of these techniques with X-Ray computed tomography
(CT) or a combination of these techniques with X-Ray computed
tomography (CT) or a combination of these techniques with X-Ray
computed tomography (CT) or a combination of these techniques with
X-Ray computed tomography (CT) or one combination of these
techniques with X-Ray computed tomography (CT) or a combination of
these techniques with X-Ray computed tomography (CT) or SPECT
imaging may be calculated using standard radiopharmaceutical
dosimetry and tissue density tables, see, Bolch et al., J. Nucl.
Med., 50(3), 477 (2009), which is incorporated by reference herein
for its teachings thereof.
[0167] In some aspects, the concentration of the one or more active
drug substances delivered to one or more susceptible tumors is
about 0.5% to about 50% of the initial dosage, including each
integer within the specified range. In some aspects, the
concentration of the one or more active drug substances delivered
to one or more susceptible tumors is about 0.5% to about 25% of the
initial dosage, including each integer within the specified range.
In some aspects, the concentration of the one or more active drug
substances delivered to one or more susceptible tumors is about
0.5% to about 15% of the initial dosage, including each integer
within the specified range. In some aspects, the concentration of
the one or more active drug substances delivered to one or more
susceptible tumors is about 0.5% to about 10% of the initial
dosage, including each integer within the specified range. In some
aspects, the concentration of the one or more active drug
substances delivered to one or more susceptible tumors is about
0.5% to about 5% of the initial dosage, including each integer
within the specified range. In some aspects, the concentration of
the one or more active drug substances delivered to one or more
susceptible tumors is about 0.5%, about 3%, about 4%, about 5%,
about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about
20%, about 25%, about 30%, about 35%, about 40%, about 45%, about
50%, about 55%, about 60%, about 65%, about 70%, or about 75% of
the initial dosage.
[0168] Assessing the percent of a delivered agent to one or more
tumors may be assessed by non-invasive techniques such as PET or
SPECT or a combination of these techniques with X-CT or MRI as
described herein. The percent absorbed initial dose delivered to a
tumor tissue measured initially using either PET or SPECT imaging
may be calculated using standard radiopharmaceutical dosimetry and
tissue density tables as described herein (e.g., as described for
delivery to a lymphatic tissue).
[0169] Alternatively, to assess relative tumor drug concentration,
one or more discovered tumors that have been treated or
administered one or more labelled (e.g., radiolabelled) agents as
described herein may be harvested from a subject. To determine the
biodistribution and percent of an initial dose delivered per gram
of tumor tissue, the specific radioactivity counts within that
tissue may be measured and quantified using standard well known
techniques and compared to the radioactivity measurements of the
initial dosage.
[0170] In some embodiments described herein are methods for methods
for increasing the amount of a bioactive agent delivered to one or
more susceptible tumors. In some aspects, because more of the
bioactive agent is targeted to the tumor, there is a smaller chance
of incurring a deleterious side effect, while exhibiting increased
therapeutic efficacy. In some aspects, the amount of bioactive
agent required to treat one or more susceptible tumors is
approximately 1% to about 75% of the dose of the identical
bioactive agent required for treating one or susceptible tumors by
conventional delivery routes; e.g., i.v., s.c., i.m., or i.d.
injections, including each integer within the specified range. In
some aspects, the amount of bioactive agent required to treat one
or more susceptible tumors is approximately 1% to about 75% of the
dose required for treating one or susceptible tumors by
conventional delivery routes; e.g., i.v., s.c., i.m., or i.d.
injections, including each integer within the specified range. In
some aspects, the amount of bioactive agent required to treat one
or more susceptible tumors is approximately 1% to about 50% of the
dose required for treating one or susceptible tumors by
conventional delivery routes; e.g., i.v., s.c., i.m., or i.d.
injections, including each integer within the specified range. In
some aspects, the amount of bioactive agent required to treat one
or more susceptible tumors is approximately 1% to about 25% of the
dose required for treating one or susceptible tumors by
conventional delivery routes; e.g., i.v., s.c., i.m., or i.d.
injections, including each integer within the specified range. In
some aspects, the amount of bioactive agent required to treat one
or more susceptible tumors is approximately 1% to about 10% of the
dose required for treating one or susceptible tumors by
conventional delivery routes; e.g., i.v., s.c., i.m., or i.d.
injections, including each integer within the specified range. In
some aspects, the amount of bioactive agent required to treat one
or more susceptible tumors is approximately about 1%, about 2%,
about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about
9%, about 10%, about 15%, about 20%, about 25%, about 30%, about
35%, about 40%, about 45%, about 50%, about 55%, about 60%, about
65%, about 70%, or about 75% of the dose required for treating one
or susceptible tumors by conventional delivery routes; e.g., i.v.,
s.c., i.m., or i.d. injections.
[0171] In some embodiments, the therapeutic efficacy of the one or
more bioactive agents delivered to one or more susceptible tumors
as described herein may be measured as a reduction in tumor size,
decreased tumor metastasis, improvement in organ tissue function,
reduction in associated side effects, reduced need for surgical
intervention, improved quality of life, increased overall survival
and increased refractory free survival or a mixture or combination
thereof.
[0172] In some embodiments, the methods of targeted delivery of one
or more bioactive agents to one or more susceptible tumors results
in a greater decrease in size of one or more tumors or reduction in
metastasis compared to conventional delivery routes; e.g., i.v.,
s.c., i.m., or i.d. injections. In some aspects, the size of the
one or more tumors is reduced by about 5% to about 99% or more,
including each integer within the specified range. In some aspects,
the size of the one or more tumors is reduced by about 5%, about
6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%,
about 25%, about 30%, about 35%, about 40%, about 45%, about 50%,
about 55%, about 60%, about 65%, about 70%, about 75%, about 80%,
about 85%, about 90% or about 95%, or about 99% or more.
[0173] Reductions in tumor size and reductions in metastasis (e.g.,
a solid tumor), and refractory free survival may be assessed and
quantified by any common imaging method known in the art. For
example magnetic resonance imaging (MRI) or magnetic resonance
spectroscopy (MRS) may be used to non-invasively track and monitor
tumor size or tumor metastasis or aggressiveness following
administration of one or more bioactive agents. See, for example,
Dynamic Contrast-Enhanced Magnetic Resonance Imaging in Oncology
(Jackson et al. eds., 2013) and Gillies and Morse., Annu. Rev.
Biomed Eng., 7, 287-326 (2005), each of which is incorporated by
reference herein for their respective teachings of MRI and MRS.
[0174] In some aspects described herein, the targeted delivery of
one or more bioactive agents to one or more susceptible tumors
results in decreased side effects. The reduction in side effects is
due to the localization of one or more bioactive agents within a
tumor or tumor metastasis. Furthermore, as described herein, a
reduction in the amount of bioactive agent required to elicit the
same therapeutic response (e.g., reduction in tumor size or
reduction in metastatic potential) decreases the amount of
potential side effects of any suitable delivered bioactive
agent.
[0175] Therefore, the methods described herein provide for the
targeted delivery of one or more bioactive agents to one or more
susceptible tumors in a subject by the initial delivery to the skin
with the methods described herein, results in previously unattained
beneficial therapeutic outcomes, comprising dose sparing, increased
drug efficacy, reduced side effects, reduced metastatic potential,
reduced tumor associated inflammation, and prolonged survival of a
subject. Accordingly, the methods described herein provide for
increased deposition of therapeutic agents within one or more
susceptible tumors when compared to i.v., s.c., i.m., or i.d.
injection methods. Any suitable cancer, tumor or metastatic
dissemination thereof may be treated by the methods described
herein. Thus, the methods described herein provide for the
treatment of a disease in a patient in need of treatment thereof;
e.g., cancer, by improving the amount of the agent deposited within
or in proximity of a tumor tissue.
[0176] In some embodiments the diffusion or movement of an
administered agent through the epidermis may be increased by
administering or contacting the epidermis of a subject with one or
more permeability or penetration enhancers. In some aspects, the
permeability or penetration enhancer may be chemical, physical, or
electrical. The permeability enhancers function to increase the
movement or diffusion of one or more agents through the stratum
corneum of the epidermis and into to the viable epidermis. The
permeability enhancers may further promote the movement or
diffusion of one or more administered agents through the viable
epidermis including the basement membrane of the viable epidermis
and into the underlying viable dermis. See, for example, Prasunitz
and Langer, Nature Biotechnol, 26(11), 1261-1268 (2008), which is
incorporated by reference herein for its teachings of the use of
epidermal permeability enhancers in transdermal drug delivery.
[0177] In some embodiments described herein, an effective amount of
one or more chemical permeability enhancers may be delivered to the
epidermis. Without being bound by any theory, it is thought that
the chemical permeability enhancers described herein may promote
the permeability of the stratum corneum to an administered agent by
denaturing intracellular keratin, causing swelling due to
hydration, affect desmosomes maintaining corneocyte adhesion, or
modify barrier producing lipids within the lipid bilayer.
Non-limiting examples of chemical permeability enhancers may
include sulfoxides, such as dimethyl sulfoxide and dodecyl methyl
sulfoxide; ureas; alcohols, such as ethanol, caprylic alcohol, and
propylene glycol; pyrrolidones and derivatives, such as
N-methyl-2-pyrrolidone and 2-pyrrolidone; azone and derivatives,
such as 1-dodecylazacycloheptan-2-one; dioxolane derivatives;
anionic, cationic, nonionic, or zwitterionic surfactants, such as
sodium lauryl sulfate, cetyltrimethyl ammonium bromide, sorbitan
monolaurate, polysorbate 80, dodecyl dimethyl ammoniopropane
sulfate; terpenes, such as menthol or limonene; fatty acids, such
as oleic acid or undecanoic acid; or hydrative amounts of
water.
[0178] In some embodiments described herein, a physical
permeability enhancer may be used to increase the permeability of
the epidermis to an administered agent. In some aspects, the
physical permeability enhancer may rely on using sound, the
application of electric fields, or specific structural interaction
with the epidermis to increase the permeability of the epidermis.
Non-limiting examples include sonophoresis (e.g., ultra sound),
iontophoresis, electroporation, or nanostructured contact surfaces
(e.g., nanotopograhy).
[0179] In some embodiments described herein, the methods for
controlled delivery of one or more agents initially to skin for the
targeted delivery to one or more tumors as described herein may
comprise delivering one or more agents through a device comprising
2 or more delivery structures that are capable of penetrating the
stratum corneum and obtaining a delivery depth and volume in the
skin and controllably delivering one or more agents at the
administration rates as described herein. The delivery structures
may be attached to a backing substrate of the delivery device and
arranged at one or a plurality of different angles for penetrating
the stratum corneum and delivering the one or more agents. In some
aspects, described herein the backing substrate comprising the
delivery structures may be in contact with the skin of a subject
and may have a cylindrical, rectangular, or geometrically irregular
shape. The backing substrate further comprises a two dimensional
surface area. In some aspects the two dimensional surface area may
be from about 1 mm.sup.2 to about 10,000 mm.sup.2. In some aspects,
the delivery structures may comprise any geometric shape (e.g., a
cylindrical, rectangular or geometrically irregular shape). In
addition, the delivery structures may comprise a length and cross
sectional surface area. In some aspects, the delivery structures
may have an overall length that is greater than a cross sectional
diameter or width. In some other aspects, the delivery structures
may have a cross sectional diameter or width greater than an
overall length. In some aspects, the cross sectional width of each
of the delivery structures may be from about 5 .mu.m to about 140
.mu.m and the cross sectional area may be from about 25 .mu.m.sup.2
to about 15,000 .mu.m.sup.2, including each integer within the
specified range. In some aspects, the length of each of the
delivery structures may be from about 10 .mu.m to about 1,000
.mu.m, including each integer within the specified range. The
surface area and cross-sectional surface areas as described herein
may be determined using standard geometric calculations known in
the art.
[0180] The delivery structures described herein need not be
identical to one another. A device having a plurality of delivery
structures may each have various lengths, outer diameters, inner
diameters, cross-sectional shapes, nanotopography surfaces, and/or
spacing between each of the delivery structures. For example, the
delivery structures may be spaced apart in a uniform manner, such
as, for example, in a rectangular or square grid or in concentric
circles. The spacing may depend on numerous factors, including
height and width of the delivery structures, as well as the amount
and type of an agent that is intended to be delivered through the
delivery structures. In some aspects, the spacing between each
delivery structure may be from about 1 .mu.m to about 800 .mu.m,
including each integer within the specified range.
[0181] In some embodiments, the delivery structures may comprise an
array of needles in fluid connection with a liquid carrier vehicle
comprising one or more agents. In some aspects, the array of
needles may comprise between 2 and 50,000 needles with structural
means for controlling skin penetration and fluid delivery to the
skin (e.g., penetrating and delivering to the skin), see, for
example, US 20150367117, which is incorporated by reference herein
in its entirety. In some aspects, the array of needles may comprise
a plurality of needles with structural means for controlling skin
penetration and fluid delivery to the skin. In some other aspects,
the array of needles may further comprise a manufactured random or
structured nanotopography on each needle. The needle or needle
array may be attached to a larger drug delivery apparatus
comprising fluidic delivery rate controls, adhesives for attaching
to the skin, fluidic pumps, and the like. If desired, the rate of
delivery of the agent may be variably controlled by the
pressure-generating means. Desired delivery rates as described
herein to the epidermis may be initiated by driving the one or more
agents described herein with the application of pressure or other
driving means, including pumps, syringes, pens, elastomer
membranes, gas pressure, piezoelectric, electromotive,
electromagnetic or osmotic pumping, or use of rate control
membranes or combinations thereof. Particular exemplary structures
and devices comprising a means for controllably delivering one or
more agents to the epidermis are described in US20110270221,
US20120187814, US20130144217, US20130144257, US20130150822,
US20130158505, US20130165861, US 20140343532, US 20150360018, US
20150367117, and US 20160106965, each of which is incorporated by
reference herein in their entirety.
[0182] In some embodiments described herein, the delivery device
may comprise a needle array in the form of a patch. In some
aspects, the array of needles are able to penetrate a most
superficial layer of the stratum corneum and initially deliver one
or more agents as described herein to at least a portion or all of
the non-viable epidermis, at least a portion of or all of the
viable epidermis, and/or at least a portion of the viable dermis of
a subject and subsequently to one or more tumors. These needles may
further comprise nanotopography on the surface of the needle in a
random or organized pattern. In some aspects, the nanotopography
pattern may demonstrate fractal geometry.
[0183] Exemplary and non-limiting devices and structures for
delivering one or more agents to the skin are shown in FIGS. 3 and
4. As shown in FIG. 3, the needle assembly as illustrated may
include a support 42 having a top surface 44 and a bottom surface
46 and defining a plurality of apertures 50 between the top and
bottom surfaces 44, 46. In addition, the needle assembly may also
include a plurality of needles 48 extending outwardly from the
bottom surface 46. As described above, each needle 48 may define
one or more channel(s) 56 in fluid communication with the apertures
50. As such, the active formulation in a liquid carrier as
described herein contained within the suitable reservoir may be
directed from the top surface 44 of the support 42 through the
apertures 50 and into the needles 48 for subsequent delivery to the
user's skin. The methods described herein, provide for the delivery
of one or more agents to the layer of the skin and ultimately to a
susceptible lymphatic capillary plexus and/or a blood capillary
plexus.
[0184] The needles described above my further comprise
nanotopography as described herein. FIG. 4 schematically
illustrates the ends of two representative needles 22. In this
particular embodiment, the needles 22 define a central exit lumen
24 that may be used for delivery of an agent via each needle 22 of
a needle array as described herein. In some other embodiments, the
needles may have multiple exit lumens for the delivery of an agent
via the needle. The surface 25 of the needle 22 may define a
nanotopography area 26. In this particular embodiment, the
nanotopography 26 defines a random pattern on the surface 25 of the
needle 22; however, in some other embodiments the nanotopography
may be structured or in a partially structured/unstructured
manner.
[0185] In some embodiments described herein, delivery devices
comprising a needle array with nanotopography as described herein
function as a permeability enhancer and may increase the delivery
of one or more agents through the epidermis. As described herein,
this delivery may occur through modulating transcellular transport
mechanisms (e.g., active or passive mechanisms) or through
paracellular permeation. Without being bound by any theory, the
nanostructured or nanotopography surface may increase the
permeability of one or more layers of the viable epidermis,
including the epidermal basement membrane by modifying cell/cell
tight junctions allowing for paracellular or modifying cellular
active transport pathways (e.g., transcellular transport) allowing
for diffusion or movement and/or active transport of an
administered agent through the viable epidermis and into the
underlying viable dermis. This effect may be due to modulation of
gene expression of the cell/cell tight junction proteins. As
previously mentioned, tight junctions are found within the viable
skin and in particular the viable epidermis. The opening of the
tight junctions may provide a paracellular route for improved
delivery of any agent, such as those that have previously been
blocked from delivery through the skin.
[0186] In some embodiments described herein, delivery devices
comprising a needle array with nanotopograhy modulate the gene
nucleic acid expression of a cell/cell contact gene of one or more
viable epithelial cell types (e.g., a viable epidermal, dermal skin
cell, blood capillary cell or lymphatic capillary cell). In some
aspects, the nucleic acid gene expression of one or more cell/cell
contact proteins is increased. In some aspects, the nucleic acid
gene expression of one or more cell/cell contact proteins is
decreased. Any method for measuring gene expression levels may be
used including but not limited to, PCR, RT-PCR, qRT-PCR,
microarrays, northern blotting, RNA Seq, and the like.
[0187] Interaction between individual cells and structures of the
nanotopography may increase the permeability of an epithelial
tissue (e.g., the epidermis) and induce the passage of an agent
through a barrier cell and encourage transcellular transport. For
instance, interaction with keratinocytes of the viable epidermis
may encourage the partitioning of an agent into the keratinocytes
(e.g., transcellular transport), followed by diffusion through the
cells and across the lipid bilayer again. In addition, interaction
of the nanotopography structure and the corneocytes of the stratum
corneum may induce changes within the barrier lipids or
corneodesmosomes resulting in diffusion of the agent through the
stratum corneum into the underlying viable epidermal layers. While
an agent may cross a barrier according to paracellular and
transcellular routes, the predominant transport path may vary
depending upon the nature of the agent.
[0188] In some embodiments described herein, delivery devices
comprising a needle array with nanotopography modulate the protein
expression of a cell/cell contact gene of one or more viable
epithelial cell types (e.g., a viable epidermal, dermal skin cell,
blood capillary cell or lymphatic capillary cell). In some aspects,
the protein expression of one or more cell/cell contact proteins is
increased. In some aspects, the protein expression of one or more
cell/cell contact proteins is decreased. Any method for measuring
protein expression levels may be used including but not limited to
western blotting, tissue imaging (e.g., fluorescent or
chemiluminescent imaging), mass spectrometry, and the like.
[0189] In some embodiments described herein, the device may
interact with one or more components of the epithelial tissue to
increase porosity of the tissue making it susceptible to
paracellular and/or transcellular transport mechanisms. Epithelial
tissue is one of the primary tissue types of the body. Epithelial
tissues that may be rendered more porous may include both simple
and stratified epithelium, including both keratinized epithelium
and transitional epithelium. In addition, epithelial tissue
encompassed herein may include any cell types of an epithelial
layer including, without limitation, keratinocytes, endothelial
cells, lymphatic endothelial cells, squamous cells, columnar cells,
cuboidal cells and pseudostratified cells. Any method for measuring
porosity may be used including but not limited to any epithelial
permeability assay. For example, a whole mount permeability assay
may be used to measure epithelial (e.g., skin) porosity or barrier
function in vivo. In one embodiment, a whole mount permeability
assay uses 5-bromo-4-chloro-3-indolyl-.beta., D-galactopyranoside
(X-Gal). Unfixed, untreated samples are rinsed with phosphate
buffered saline (PBS) and briefly dried. Samples are immersed in a
standard X-Gal reaction mixture with the pH adjusted to 4.5. After
incubating at 37.degree. C. for 8-10 hrs, the samples are washed
with PBS for 1-2 minutes and analyzed. In one embodiment, a whole
mount permeability assay uses a histological dye such as, but not
limited to, toluidine blue or hematoxylin. Unfixed, untreated
samples are incubated for 1-5 minutes in methanol and rinsed in
PBS. Samples are incubated in 0.5% hematoxylin or 0.1% toluidine
blue then embedded in agarose for analysis. In one embodiment,
sample analysis is performed by photographing the prepared samples
and evaluated based on the degree of dye penetration. Other methods
as are known in the art may also be used. See, for example, Indra
and Leid., Methods Mol Biol., (763) 73-81 (2012), which is
incorporated by reference herein for its teachings thereof.
[0190] In some embodiments described herein, the structural changes
induced by the presence of a nanotopography surface on a barrier
cell are temporary and reversible. It was surprisingly found that
using nanostructured nanotopography surfaces results in a temporary
and completely reversible increase in the porosity of epithelial
tissues by changing junctional stability and dynamics, which
without being bound by any theory, may result in a temporary
increase in the paracellular and transcellular transport of an
administered agent through the epidermis and into the viable
dermis. Thus, in some aspects, the increase in permeability of the
epidermis or an epithelial tissue elicited by the nanotopography,
such as promotion of paracellular or transcellular diffusion or
movement of one or more agents, returns to a normal physiological
state that was present before contacting the epithelial tissue with
a nanotopography following the removal of the nanotopography. In
this way, the normal barrier function of the barrier cell(s) (e.g.,
epidermal cell(s)) is restored and no further diffusion or movement
of molecules occurs beyond the normal physiological diffusion or
movement of molecules within the tissue of a subject.
[0191] These reversible structural changes induced by the
nanotopography may function to limit secondary skin infections,
absorption of harmful toxins, and limit irritation of the dermis.
Also the progressive reversal of epidermal permeability from the
top layer of the epidermis to the basal layer may promote the
downward movement of one or more agents through the epidermis and
into the dermis and prevent back flow or back diffusion of the one
or more agents back into the epidermis.
[0192] In some alternative embodiments, the methods for delivering
one or more agents to the skin comprises not only a needle,
microneedle, or nanoneedle-based injection means, but other
delivery methods, such as needle-less or needle-free ballistic
injection of fluids, iontophoresis techniques, and direct
deposition of fluid, solids, or other dosing forms into the
epidermis of the skin.
[0193] In some embodiments described herein, are methods for
applying a device having at least 2 or more delivery structures to
the surface of the skin a subject for the treatment of a disease or
disorder described herein. In some aspects, the device is applied
to an area of the subject's skin, wherein the location of the skin
on the body is dense in lymphatic capillaries and/or blood
capillaries. Multiple devices may be applied to one or more
locations of the skin having a dense network of lymphatic
capillaries. In some aspects, 1, 2, 3, 4, 5, or more devices may be
applied. These devices may be applied spatially separate or in
close proximity or juxtaposed with one another. Exemplary and
non-limiting locations dense with lymphatics comprise the palmar
surfaces of the hands, the scrotum, the plantar surfaces of the
feet and the lower abdomen.
[0194] It will be readily apparent to one of ordinary skill in the
relevant arts that suitable modifications and adaptations to the
compositions, methods, and applications described herein can be
made without departing from the scope of any embodiments or aspects
thereof. The compositions and methods provided are exemplary and
are not intended to limit the scope of the specified embodiments.
All of the various embodiments, aspects, and options disclosed
herein can be combined in all variations. The scope of the
compositions, formulations, methods, and processes described herein
include all actual or potential combinations of embodiments,
aspects, options, examples, and preferences herein described. All
patents and publications cited herein are incorporated by reference
herein for the specific teachings thereof.
Example 1. Diagram of the Skin
[0195] The overall structure of the skin including the dermis and
epidermis is illustrated in FIG. 1. The dermis is composed of a
myriad of tissue types, and in general, exhibits an overall
thickness ranging from about 500 .mu.m to about 4,000 .mu.m. The
lymphatic and blood capillaries are found often found together as
illustrated or they can be present as separate entities. As
illustrated, the blood and lymphatic capillaries are often located
within the upper portions of the dermis (e.g., near the epidermal
dermal or epidermal basement mem brane) within a portion of the
papillary dermis. Larger vessels are generally found within the
lower reticular dermis (e.g., a blood vessel as shown). Other
tissue types important to dermal function include the larger
arteries, arterioles, sweat gland ducts, sebaceous glands, nerve
corpuscles, connective tissues and extra cellular matricis, smooth
muscle, and hair follicles. Below the reticular dermis lies the
subcutaneous tissue layer, which is composed largely of fat tissue
and generally is void of any lymphatic or blood vasculature.
[0196] As illustrated in FIG. 2A, the epithelial skin layer is
formed of four principal cellular layers lacking the many other
tissue types of the dermis (e.g., blood and lymphatic capillaries,
etc.) with a general thickness ranging from about 20 .mu.m to about
400 .mu.m. As illustrated from top to bottom is the basement
membrane followed by the basal layer or stratum germinativum, the
squamous cell layer or the stratum spinosum (spinous layer), the
granular cell layer or the stratum granulosum, and the comified
layer or the stratum corneum. The epidermis is principally
non-mitotic with the stratum corneum comprising non-viable
enucleated barrier providing cells; however, as illustrated in FIG.
2B the basal layer consists of symmetrically dividing stem cells
and other transiently amplifying cells for the regeneration of the
corneum.
Example 2. Depth of Penetration into the Skin
[0197] An array of needles was fabricated on a patch and was used
to estimate the average range of depth of delivering of an agent
within the skin of adult guinea pigs. As shown in FIGS. 5A, 5B, and
5C, methylene blue dye was administered to an average depth of
about 92 .mu.m demonstrating a range of depth distributions of
about 5 .mu.m to about 200 .mu.m (a Gaussian distribution of
depths). As shown in FIG. 6, the structure and depth in the skin
may be estimated by using optical coherence tomography techniques.
The structure of the skin after applying an array of needles can be
visualized by looking at individual horizontal slices of the
skin.
Example 3. Modulation of Epidermal Tight Junction Proteins
[0198] An array of needles having a nanotopography surface was
fabricated on a patch and tested on an in vitro mono layer of
Caco-2 epithelial cells. As shown in FIG. 7A, the ZO-1 tight
junction protein shows a normal staining pattern. However, when an
array of needles having a nanotopograpby surface is placed within
proximity of Caco-2 cells, a disrupted staining pattern can be
visualized. This ruffled pattern indicates junction remodeling in
the areas of where the nanotopography was located (FIG. 7B). When
the array of needles having a nanotopography is removed, the
staining pattern returns to normal, indicating a spatial and
temporal effect on tight junction proteins, such as ZO-1 (FIG.
7C).
Example 4. Better Delivery of Trastuzumab (Herceptin.RTM.) to
Tumors In Vivo
[0199] The ability to deliver trastuzumab (an anti-cancer drug) to
tumors via administration to the skin was tested in a mouse
xenograft tumor model. The HER-2 positive JIMT-1 human breast
cancer cell line was used to generate xenograft tumors in mice. An
array of needles having a nanotopography surface was fabricated on
a patch and applied to the dorsal surface of mice presenting with
tumors. These mice were then administered different amounts of
trastuzumab at different administration rates to the skin and the
concentration of trastuzumab in tumors was assessed. As shown in
FIG. 8, administration of 0.22 mg of trastuzumab at a rate of 100
.mu.1/hr yielded higher concentrations of trastuzumab in JIMT-1
tumors compared to approximately a 10 fold greater dosage of
trastuzumab administered intravenously (2 mg i.v.).
[0200] Trastuzumab administered to the skin demonstrated a greater
or equivalent efficacy in treating tumors as evidenced by large
areas of tumor necrosis (black arrows pointing to sections of
necrotic tissue) when compared to even higher doses of trastuzumab
administered intravenously (FIGS. 9A-B).
Example 5. Better Delivery of Drugs to Tumors In Vitro
[0201] The efficiency of directly delivering drugs within in vitro
grown tumors using an array of needles having a nanotopography
surface was tested. Tumor tissues were grown in vitro and
administered an anti-cancer drug by either adding drug to the
tissue culture media or administering with an array of needles. The
distribution of the drug was assessed by cryosectioning and
subsequent tissue visualization (FIGS. 10A and 10B). The effects on
cancer cell proliferation were measured using standard
proliferation assay staining techniques following drug delivery. As
shown in FIG. 10A, simple addition of the drug to the culture media
led to little drug absorption throughout the tumor with the
majority being retained within cells at the surface layer (as shown
by the bracket). In contrast, as shown in FIG. 10B, administration
of the drug to the tumor with an array of needles demonstrated
higher levels of distribution throughout the tumor tissue slice
with little being retained at the surface (as shown by black arrow
head). Drugs that were only supplemented with the tissue culture
media had a reduced anti-proliferative effect (FIG. 10C) compared
to direct administration using an array of needles (FIG. 10D
examples of proliferating cells indicated by arrow heads).
Example 6. In Vivo Imaging of Lymphatic Delivery of Etanercept
(Enbrel.RTM.--an Anti-Inflammatory Drug) to the Lymphatic
Vasculature and Biodistribution in Rats
[0202] The ability to deliver the protein therapeutic Etanercept
directly to the lymphatic system via administration to the skin was
tested. Etanercept was fluorescently tagged for in vivo
visualization using near infrared light as previous described, see
Sevick-Muraca et al., J. Clin Invest., 124(3), 905-914 (2014),
which is incorporated by reference herein for its teachings
thereof. Etanercept was administered to the skin of rats by placing
a delivery device comprising an array of needles dorsally on the
rats. As shown in FIG. 11 and FIG. 12, administration of etanercept
to the skin resulted in uptake by the lymphatic vasculature and
subsequent distribution to primary and secondary lymph node
tissues.
[0203] The biodistribution of etanercept across multiple tissues
types following delivery was investigated. Accordingly, delivering
etanercept to the skin resulted in much higher levels within the
axillary and inguinal lymph nodes when compared to traditional
i.v., s.c., or i.d. methods (FIG. 13).
Example 7. Rate of Blood Serum Absorption of Etanercept after
Delivery to the Skin
[0204] As shown in FIG. 14, the blood serum absorption rate of
etaoercept following administration to the skin is approximately
the same as i.v., s.c., or i.d. methods.
* * * * *