U.S. patent application number 13/489324 was filed with the patent office on 2012-11-22 for needle array assembly and method for delivering therapeutic agents.
Invention is credited to S. Bahram Bahrami, James Olson, Mandana Veiseh.
Application Number | 20120296206 13/489324 |
Document ID | / |
Family ID | 40351485 |
Filed Date | 2012-11-22 |
United States Patent
Application |
20120296206 |
Kind Code |
A1 |
Bahrami; S. Bahram ; et
al. |
November 22, 2012 |
NEEDLE ARRAY ASSEMBLY AND METHOD FOR DELIVERING THERAPEUTIC
AGENTS
Abstract
A fluid delivery device includes an array of needles, each in
fluid communication with a respective reservoir. Respective
actuators are coupled so as to be operable to drive fluid from the
reservoirs via needle ports. Each needle can have a plurality of
ports, and the ports can be arranged to deliver a substantially
equal amount of fluid at any given location along its length. A
driver is coupled to the actuators to selectively control the rate,
volume, and direction of flow of liquid through the needles. The
device can simultaneously deliver a plurality of fluid agents along
respective axes in solid tissue in vivo. If thereafter resected,
the tissue can be sectioned for evaluation of an effect of each
agent on the tissue, and based on the evaluation, candidate agents
selected or deselected for clinical trials or therapy, and subjects
selected or deselected for clinical trials or therapeutic
treatment.
Inventors: |
Bahrami; S. Bahram;
(Emeryville, CA) ; Veiseh; Mandana; (Emeryville,
CA) ; Olson; James; (Seattle, WA) |
Family ID: |
40351485 |
Appl. No.: |
13/489324 |
Filed: |
June 5, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12674146 |
Aug 27, 2010 |
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PCT/US2008/073212 |
Aug 14, 2008 |
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13489324 |
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60955676 |
Aug 14, 2007 |
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Current U.S.
Class: |
600/431 ;
600/407; 600/562; 604/173; 604/65; 604/93.01 |
Current CPC
Class: |
A61M 5/3294 20130101;
A61M 5/3298 20130101; A61M 5/3291 20130101; A61M 5/16827 20130101;
A61M 5/3287 20130101; A61M 2205/50 20130101; A61M 5/14526 20130101;
A61M 5/1452 20130101; A61M 5/20 20130101; A61B 6/508 20130101; A61P
35/00 20180101; A61M 2205/52 20130101 |
Class at
Publication: |
600/431 ;
604/93.01; 600/562; 604/65; 604/173; 600/407 |
International
Class: |
A61B 6/00 20060101
A61B006/00; A61B 10/02 20060101 A61B010/02; A61M 5/00 20060101
A61M005/00 |
Claims
1.-121. (canceled)
122. An array, comprising: an array of reservoirs and means for
administering a therapeutic or diagnostic agent in tissue
immediately adjacent to the administration means and separate from
the other administration means.
123. The array of claim 122, further comprising means for
separately extracting, sampling or analyzing the tissue or cells at
the sites of administration.
124. The array of claim 122, wherein the array provides for
separate release of agent in the separate reservoirs.
125. The array of claim 122, wherein each of the reservoirs may be
loaded with different substances and/or different microdose amounts
of the substances, which can be released independently, optionally
further comprising means for controlling release selected from the
group consisting of a microprocessor, remote control and a
sensor.
126. The array of claim 122, wherein the reservoirs comprise
biodegradable polymeric matrices which release agent as a function
of diffusion or degradation.
127. The array of claim 122 providing microdoses of therapeutic or
diagnostic agents.
128.-129. (canceled)
130. The array of claim 122, wherein the means for extracting,
sampling or analyzing the tissue or cells are biopsy
mechanisms.
131. (canceled)
132. The array of claim 122, comprising penetrating members.
133. The array of claim 122, comprising an assay component
configured to detect and analyze genes or their products which are
over-expressed or over-active in cells undergoing unwanted
proliferation, or to detect abnormal cellular proliferation.
134. The array of claim 122, comprising imaging means.
135.-151. (canceled)
152. A device, comprising a plurality of reservoirs and a plurality
of needles arranged in an array for delivering one or more fluid
agents to a plurality of positions along mutually parallel axes
within a tissue of a subject.
153. The device of claim 152, further comprising means for
separately obtaining or analyzing biological samples at the sites
of administration.
154. The device of claim 152, wherein each one of said plurality of
reservoirs is in fluid communication with a respective one of said
plurality of needles, allowing separate release of said one or more
fluid agents in separate reservoirs.
155. The device of claim 152, wherein each one of said plurality of
reservoirs is loaded with different fluid agents and/or different
dosage of said agents, which can be released independently,
optionally further comprising means for controlling release
selected from the group consisting of a microprocessor, a
controller and a patient scanning device.
156. The device of claim 152, wherein said plurality of reservoirs
comprise biodegradable polymeric carrier to control the release of
said one or more fluid agents.
157. The device of claim 152, providing an amount of said one or
more fluid agents to be locally delivered to said tissue.
158.-159. (canceled)
160. The device of claim 152, wherein the means for obtaining or
analyzing biological samples are biopsy mechanisms.
161. (canceled)
162. The device of claim 152, comprising an assay component
configured to detect and analyze an altered physiologic state
include cell viability, cell division, apoptosis, necrosis, cell
surface marker expression and cellular activation state.
163. The device of claim 152, further comprising imaging means.
164. (canceled)
165. A device, comprising an array of needles for separately
obtaining or analyzing biological samples from different sites of a
tissue within an individual.
166.-179. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application is a national stage application of
PCT/US2008/73212, filed Aug. 14, 2008, which claims the benefit
under 35 U.S.C. .sctn.119(e) of U.S. Provisional Patent Application
No. 60/955,676 filed Aug. 14, 2007. The contents of each of the
above-named documents are incorporated herein by reference for all
purpose in their entirety.
BACKGROUND
[0002] 1. Technical Field
[0003] In general, the disclosed embodiments relate to devices and
methods for the introduction and subsequent evaluation of
therapeutic agents to biological tissue, and in particular to the
simultaneous introduction of a plurality of agents to the tissue in
vivo.
[0004] 2. Description of the Related Art
[0005] Numerous cancer-related therapeutics are under phase I or
phase II clinical trial and evaluations at any particular time;
however, most of them will fail to advance. In fact, it is
estimated that more than 90% of cancer-related therapeutics will
fail phase I or II clinical trial evaluation. The failure rate in
phase III trials is almost 50%, and the cost of new drug
development from discovery through phase III trials is between $0.8
billion and $1.7 billion and can take between eight and ten
years.
[0006] In addition, many patients fail to respond even to standard
drugs that have been shown to be efficacious. For reasons that are
not currently well understood or easily evaluated, individual
patients may not respond to standard drug therapy. One significant
challenge in the field of oncology is to exclude drug selection for
individual patients having cell autonomous resistance to a
candidate drug to reduce the risk of unnecessary side effects. A
related problem is that excessive systemic concentrations are
required for many oncology drug candidates in efforts to achieve a
desired concentration at a tumor site, an issue compounded by poor
drug penetration in many under-vascularized tumors (Tunggal et al.,
1999 Clin. Canc. Res. 5:1583).
[0007] Clearly there is a need in the art for improved devices and
methods for testing and delivering cancer therapies, including
improved methodologies for performing efficient pre-clinical and
clinical studies of candidate oncology medicines, and for
identifying therapeutics having increased likelihood of benefitting
individual subjects. The present invention addresses these and
similar needs, and offers other related advantages.
BRIEF SUMMARY
[0008] It is an aspect of the present invention to provide a device
for delivery of a fluid to a solid tissue, comprising: a plurality
of needles arranged in an array; a plurality of reservoirs, each in
fluid communication with a respective one of the plurality of
needles; and a plurality of actuators operatively coupled to
respective ones of the plurality of reservoirs and configured to
control a fluid pressure within the reservoir. In certain
embodiments each of the plurality of actuators comprises one of a
plurality of plungers, a first end of each of the plurality of
plungers being received in a respective one of the plurality of
reservoirs, and in certain further embodiments the plungers of the
plurality of plungers are operatively coupled together at
respective second ends so as to be simultaneously depressable.
Certain still further embodiments comprise a plunger driver
configured to depress all of the plurality of plungers at a
selectively variable rate. In other embodiments each of the
plurality of actuators comprises one of a plurality of fluid
transmission lines having first and second ends, a first end of
each of the plurality of fluid transmission lines being coupled to
a respective one of the plurality of reservoirs. In other
embodiments the device comprises a fluid pressure source, and each
of the plurality of actuators comprises a fluid coupling between
the fluid pressure source and a respective one of the plurality of
reservoirs. In further embodiments the fluid pressure source
comprises at least one of a compressor, a vacuum accumulator, a
peristaltic pump, a master cylinder, a microfluidic pump, and a
valve. In another embodiment, each of the plurality of needles
comprises a plurality of ports distributed along its length.
[0009] In another embodiment there is provided a device for
delivering a fluid to a solid tissue, comprising a dispenser
including a needle having a plurality of ports distributed along a
length thereof, a reservoir in fluid communication with the
dispensing needle, and a plunger having a first end positioned in
the reservoir; and a plunger driver coupled to a second end of the
plunger and configured to depress the plunger at a selectably
variable rate. In certain further embodiments the dispenser is one
of a plurality of dispensers arranged in a dispenser array, each
comprising a needle, a reservoir, and a plunger having first and
second ends. In certain further embodiments the plunger driver is
coupled to the second end of the plunger of each of the plurality
of dispensers and is configured to depress each of the plungers
simultaneously. In certain other further embodiments the device
comprises a plurality of cylindrical tubes arranged in an array
corresponding to the dispenser array, each of the plurality of
cylindrical tubes being sized and positioned to receive the needle
of a respective one of the plurality of dispensers.
[0010] In certain other embodiments the plunger driver comprises a
driver shaft coupled to the plunger and having a threaded region,
the plunger driver configured such that rotation of the driver
shaft in a first direction depresses the plunger a distance
corresponding to a thread pitch of the threaded region and a number
of revolutions of the driver shaft. In certain further embodiments
the device comprises a motor having a rotor coupled to the driver
shaft of the plunger driver such that the rotor and the driver
shaft are rotationally fixed with respect to each other, the motor
being controllable to rotate the rotor at a selectably variable
rate. In certain other further embodiments the device comprises a
motor having a rotor coupled to the driver shaft of the plunger
driver such that the rotor and the driver shaft are rotationally
fixed with respect to each other, the motor being controllable to
rotate the rotor to a selectable angle of rotation. Certain further
embodiments comprise a controller coupled to the motor, the
controller being programmable to control direction and speed of
rotation of the rotor and to control a number of degrees from a
start of rotation to an end of rotation. In other embodiments of
the above described device, the dispenser comprises a dispenser
cylinder; a first portion of the dispenser cylinder defines the
reservoir; and a second portion of the dispenser cylinder defines
the needle. In another embodiment the plurality of ports are sized
and positioned along the length of the needle so as to deliver a
substantially equal amount of fluid at any given location along the
length of the needle. In another embodiment the plurality of ports
is evenly distributed along a portion of the length of the
needle.
[0011] In certain embodiments a size of each of the plurality of
ports is inversely related to a distance of the respective port
from a tip-end of the needle. In certain other embodiments a
distribution density of the plurality of ports is inversely related
to a distance of the respective port from a tip-end of the needle.
In certain other embodiments the plurality of ports is distributed
in a spiral pattern along the length of the needle. In certain
other embodiments the plurality of ports is arranged in pairs of
ports on opposite sides of the needle, with each pair of ports
rotated 90 degrees with respect to adjacent pairs of ports along
the length of the needle.
[0012] According to certain other embodiments disclosed herein,
there is provided a method, comprising placing an agent in a
reservoir of each of a plurality of dispenser needles; inserting
each of the plurality of dispenser needles into a selected region
of solid tissue; and introducing the agent in the reservoirs into
the selected region of solid tissue by simultaneously
overpressurizing each of the plurality of dispenser needles. In
certain further embodiments the introducing comprises introducing
the agent in the reservoirs into the selected region of solid
tissue from a plurality of apertures along each of the plurality of
dispenser needles. Certain other further embodiments comprise at
least one of imaging the solid tissue prior to the inserting,
imaging the solid tissue concurrently with the inserting, and
imaging the solid tissue after the inserting. In certain other
further embodiments the inserting comprises inserting an array of
introducer needles into a subject; inserting each of the plurality
of dispenser needles into a respective one of the array of
introducer needles; and extending a tip-end of each of the
plurality of dispenser needles beyond a tip end of the respective
one of the array of introducer needles and into the selected region
of tissue. Certain further embodiments comprise removing stylets
from the introducer needles of the array prior to inserting the
plurality of dispenser needles.
[0013] In certain embodiments the selected region of tissue is a
portion of a tumor in a subject, and in certain further embodiments
the subject is one of a preclinical model and a human patient. In
certain other embodiments the method comprises excising at least
the portion of the tumor after the introducing. Certain further
embodiments comprise at least one of imaging the tumor prior to the
excising, imaging the tumor concurrently with the excising, and
imaging the tumor after to the excising. In certain other
embodiments the excising comprises excising at least the portion of
the tumor at a time that is a selected period of time after
introducing the agent. In certain further embodiments the selected
period of time is one of a range of time, a minimum period of time
for excising, and a specific period of time for excising. In
certain embodiments the selected period of time is a period
exceeding 48 hours. In certain embodiments the selected period of
time is a range of between about 72 and about 96 hours. In certain
embodiments the selected period of time is a period exceeding one
week.
[0014] According to certain other embodiments of the above
described method, the agent comprises a plurality of agents, and
the placing comprises placing each of the plurality of agents into
the reservoir of a respective one of the plurality of dispenser
needles. In certain further embodiments the plurality of agents
comprises at least one of a negative control composition and a
positive control composition. In certain other further embodiments
the plurality of agents comprises at least one position marker. In
certain other further embodiments at least one of the plurality of
agents is a candidate effective agent. In certain other further
embodiments at least one of the plurality of agents comprises an
indicator of efficacy, which in certain further embodiments
comprises at least one of a nanoparticle, a nanostucture, and an
indicator dye. In certain other embodiments at least one of the
plurality of agents is selected based on a clinically demonstrated
efficacy of the respective agent. In certain other further
embodiments of the above described method, the method comprises
assessing, with respect to at least one of the plurality of agents,
at least one of efficacy, activity, and toxicity of the agent.
[0015] In another embodiment there is provided a method for
identifying relative efficacies of a plurality of agents for
treating a subject, comprising injecting each of a plurality of
candidate effective agents into a respective location in an
injection site in a solid tissue in a subject; excising from the
subject at least the injection site of the solid tissue; and
evaluating the excised injection site for an altered physiologic
state at each of the respective locations, and therefrom
identifying relative efficacies of the plurality of agents. In
certain further embodiments the excising comprises one of excising
at least 48 hours after the injecting, excising at least 72 hours
after the injecting, excising 72 to 96 hours after the injecting,
and excising at least one week after the injecting.
[0016] In another embodiment there is provided a method of
operation of a therapeutic device, comprising charging a reservoir
of each of a plurality of needles with a respective one of a
plurality of agents; injecting, simultaneously, each of the
plurality of agents into a respective region of a solid tissue; and
evaluating an effect of each of the plurality of agents on the
respective region. In certain further embodiments the injecting
comprises injecting the plurality of agents into the solid tissue
in vivo, and in certain still further embodiments the method
comprises excising the solid tissue prior to the evaluating. In
certain embodiments the method comprises imaging the solid tissue,
which in certain further embodiments comprises imaging the solid
tissue in vivo. In certain other embodiments the injecting
comprises distributing each of the plurality of agents into the
solid tissue along an axis in the respective region of the tissue.
In certain other embodiments the method further comprises
assessing, with respect to at least one of the plurality of agents,
at least one of efficacy, activity, and toxicity of the agent.
[0017] Also provided herein according to certain embodiments is a
method of determining efficacy of a cancer treatment regimen,
comprising simultaneously introducing an agent to a plurality of
positions in a solid tumor in a subject in vivo; removing the tumor
from the subject; and evaluating an effect of the agent on the
tumor in vitro. In certain further embodiments the agent comprises
a plurality of agents and the introducing comprises distributing
each of the plurality of agents to a respective one of the
plurality of positions in the tumor. In another embodiment there is
provided a method, comprising introducing an agent to a region of
solid tissue in a subject by distributing the agent to a plurality
of positions along an axis within the region of solid tissue in
vivo; removing the region of solid tissue from the subject; and
evaluating an effect of the agent on the region of solid tissue in
vitro. In a further embodiment the region of solid tissue comprises
a tumor.
[0018] In certain embodiments the axis is one of a plurality of
parallel axes in the region of solid tissue, and wherein the
introducing comprises distributing the agent along each of the
plurality of parallel axes. In certain further embodiments the
introducing comprises simultaneously distributing the agent along
each of the plurality of parallel axes, and in certain other
further embodiments the plurality of parallel axes is arranged in
an array. In certain other embodiments the method comprises
introducing at least two position markers to the region of solid
tissue along a respective one of the plurality of parallel axes,
and in certain further embodiments the introducing at least two
position markers comprises distributing the at least two position
markers along respective parallel axes within the region of solid
tissue. In certain other embodiments the at least two position
markers each comprise a detectable label that is selected from the
group consisting of a radiolabel, a radio-opaque label, a
fluorescent label, a colorimetric label, a dye, an enzymatic label,
a GCMS tag, avidin, and biotin.
[0019] In certain other embodiments of the above described method,
the agent is one of a plurality of agents and the axis is one of a
plurality of parallel axes arranged in an array in the region of
solid tissue, and wherein the introducing comprises distributing
each of the plurality of agents to a plurality of positions along a
respective one of the plurality of parallel axes. In certain other
embodiment the method comprises at least one of imaging the solid
tissue prior to the introducing, imaging the solid tissue
concurrently with the introducing, and imaging the solid tissue
after the introducing. In certain other embodiments the evaluating
comprises sectioning the region of solid tissue into a plurality of
sections normal to the parallel axes. In certain further
embodiments the evaluating comprises detecting within the solid
tissue an altered physiologic state that results from at least one
of the plurality of agents. In certain further embodiments the
detecting comprises, with respect to the at least one of the
plurality of agents, at least one of detecting a degree of
permeation of the agent through the solid tissue, detecting a
physicochemical effect of the agent on the tissue, and detecting a
pharmacological effect of the agent on the tissue. In certain other
embodiments the evaluating comprises determining the effects of at
least two of the plurality of agents on a same position within the
region of the solid tissue. In certain other embodiments the
evaluating comprises determining the effects of at least two of the
plurality of agents on adjacent positions within the region of the
solid tissue.
[0020] In certain other embodiments the evaluating comprises
differentiating a degree of the effect of at least one of the
plurality of agents on different sections of the solid tissue
according to different characteristics of the different sections of
the solid tissue. In certain other embodiments the evaluating
comprises comparing a first effect of at least a first one of the
plurality of agents on the solid tissue with a second effect of at
least a second one of the plurality of agents on the solid tissue.
In certain other embodiments the evaluating comprises, with respect
to at least one of the plurality of agents, assessing at least one
of efficacy, activity, and toxicity on the region of solid tissue.
In certain other embodiments the method comprises deselecting at
least one of the plurality of agents based on the evaluating. In
certain other embodiments the method comprises selecting at least
one of the plurality of agents based on the evaluating. In certain
other embodiments the method comprises prioritizing at least two of
the plurality of agents based on the evaluating. In certain other
embodiments the method comprises distributing the plurality of
agents to a plurality of positions, each along a respective one of
a plurality of parallel axes within a region of solid tissue within
each of a plurality of subjects. In certain further embodiments the
method comprises one of (i) selecting at least one of the plurality
of agents based on the evaluating, (ii) deselecting at least one of
the plurality of agents based on the evaluating, and (iii)
prioritizing at least two of the plurality of agents based on the
evaluating. In certain other embodiments the method comprises one
of (i) selecting at least one of the plurality of subjects based on
the evaluating, (ii) deselecting at least one of the plurality of
subjects based on the evaluating, and (iii) prioritizing at least
two of the plurality of subjects based on the evaluating. In
certain other embodiments the evaluating comprises determining a
level of altered physiologic state of the solid tissue near at
least one of the plurality of parallel axes.
[0021] Turning to another embodiment there is provided a fluid
agent-delivering device comprising (i) a plurality of needles
arranged in an array, each of said needles having, independently,
one or a plurality of ports distributed along its length wherein at
least one needle has said plurality of ports, (ii) a plurality of
reservoirs containing the fluid agent, each of said reservoirs
being in fluid communication with a respective one of the plurality
of needles, and (iii) a plurality of plungers, a first end of each
plunger being received in a respective one of the plurality of
reservoirs and a second end of each plunger being depressable such
that depressing each plunger results in injection of the fluid
agent through the respective one of the plurality of needles.
[0022] In another embodiment of the presently disclosed invention
there is provided a method for selective delivery of a fluid agent
to a solid tissue, comprising (a) introducing a plurality of
needles of a fluid agent-delivering device into the solid tissue;
and (b) administering the fluid agent into the solid tissue by
injection through said needles. In certain further embodiments the
solid tissue has been removed from a subject. In certain other
further embodiments the solid tissue is in a subject. In certain
further embodiments the agent is delivered to the solid tissue in a
therapeutically effective amount. In certain still further
embodiments, outside the solid tissue, the agent is either (i)
undetectable, or (ii) if detectable outside the solid tissue, the
agent is present at less than a minimal dose. In certain
embodiments the solid tissue comprises a tumor. In certain further
embodiments the tumor is selected from a benign tumor and a
malignant tumor. In certain other further embodiments the tumor is
selected from a primary tumor, an invasive tumor and a metastatic
tumor. In certain other further embodiments the tumor comprises at
least one cancer cell selected from a prostate cancer cell, a
breast cancer cell, a colon cancer cell, a lung cancer cell, a
brain cancer cell, and an ovarian cancer cell. In certain other
further embodiments the tumor comprises a cancer selected from
adenoma, adenocarcinoma, squamous cell carcinoma, basal cell
carcinoma, small cell carcinoma, large cell undifferentiated
carcinoma, chondrosarcoma and fibrosarcoma. In certain other
embodiments the solid tissue is selected from brain, liver, lung,
kidney, prostate, ovary, spleen, lymph node, thyroid, pancreas,
heart, skeletal muscle, intestine, larynx, esophagus and
stomach.
[0023] In certain other embodiments the fluid agent comprises an
agent that is selected from (a) a gene therapy agent; (b) a
chemotherapy agent; (c) a small molecule; (d) an antibody; (e) a
protein; (f) one of a small interfering RNA and an encoding
polynucleotide therefor; (g) one of an antisense RNA and an
encoding polynucleotide therefor; (h) one of a ribozyme and an
encoding polynucleotide therefor; (i) a detectable label; and (j)
one of a therapeutic protein, polypeptide, and a peptidomimetic. In
certain further embodiments the detectable label is selected from a
radiolabel, a radio-opaque label, a fluorescent label, a
colorimetric label, a dye, an enzymatic label, a GCMS tag, avidin,
and biotin. In certain embodiments the agent is selected from (i) a
gene therapy agent that comprises at least one operably linked
promoter, (ii) a small interfering RNA-encoding polynucleotide that
comprises at least one operably linked promoter; (iii) an antisense
RNA-encoding polynucleotide that comprises at least one operably
linked promoter; and (iv) a ribozyme-encoding polynucleotide that
comprises at least one operably linked promoter. In certain further
embodiments the operably linked promoter is selected from a
constitutive promoter and a regulatable promoter. In certain still
further embodiments the regulatable promoter is selected from an
inducible promoter, a tightly regulated promoter and a
tissue-specific promoter.
[0024] In certain other embodiments there is provided a method for
altering a physiologic state in a solid tissue, comprising: (a)
introducing a plurality of needles of a fluid agent-delivering
device into the solid tissue; and (b) administering the fluid agent
into the solid tissue by injection through said needles.
[0025] In certain embodiments there is provided a method for
obtaining biological samples from a plurality of positions in a
solid tissue, comprising (a) introducing a multiple needle device
into the solid tissue, thereby placing a plurality of needles at a
plurality of positions in the tissue; and (b) generating negative
pressure at a port of each needle of said multiple needle device
under conditions and for a time sufficient to draw into said
needles a plurality of biological samples from said plurality of
positions in the tissue, and thereby obtaining biological samples
from a plurality of positions in the tissue.
[0026] In certain embodiments there is provided a method for
obtaining biological samples from a plurality of positions along an
axis in a solid tissue, comprising (a) introducing a multiple
needle device into the solid tissue, thereby placing a plurality of
needles at a plurality of positions in the tissue; and (b)
generating negative pressure at a plurality of ports located along
a length of each needle of said multiple needle device under
conditions and for a time sufficient to draw into said needles a
plurality of biological samples from said plurality of positions in
the tissue, and thereby obtaining biological samples from a
plurality of positions along an axis in the tissue.
[0027] In certain embodiments there is provided a method of
screening subjects for eligibility to participate in a clinical
trial of one or more agents, comprising (a) introducing one or more
agents to a region of solid tissue in one or more subjects in vivo
by distributing each of said agents to a plurality of positions
along an axis within the region in each subject; (b) removing the
region of solid tissue from each of said subjects; and (c)
evaluating each region removed in (b) for an effect of each agent
on the respective position along the axis within the region,
wherein either (i) for any given agent or agents presence of a
detectable effect of said agent or agents on the solid tissue
region from the subject indicates eligibility of the subject for
participation in a clinical trial of the agent or agents, (ii) for
any given agent or agents absence of a detectable effect of said
agent or agents on the solid tissue region from the subject
indicates ineligibility of the subject for participation in a
clinical trial of the agent or agents, or (iii) both (i) and
(ii).
[0028] In certain embodiments there is provided a method of rating
a candidate agent for development into a therapeutic agent for
treating a solid tumor, comprising (a) introducing one or more
candidate agents to a region of a solid tumor of known tumor type
in each one or more subjects having a tumor of the known tumor
type, by distributing each of said candidate agents to a plurality
of positions along an axis within the region in each subject; (b)
removing the region of solid tumor from each of said subjects; and
(c) comparing each region removed in (b) for an effect of each
candidate agent on the respective position along the axis within
the region, wherein an agent that results in a greater beneficial
effect when introduced to the tumor receives a higher rating for
development into a therapeutic agent for treating the solid tumor,
and an agent that results in a lesser beneficial effect when
introduced to the tumor receives a lower rating for development
into a therapeutic agent for treating the solid tumor.
[0029] These and other aspects of the present invention will become
apparent upon reference to the following detailed description and
attached drawings.
BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS
[0030] FIG. 1 is a schematic diagram of a needle array assembly for
injecting biological tissue with therapeutic agents according to
various embodiments.
[0031] FIGS. 2A-2D and 3 show delivery needles according to
respective embodiments.
[0032] FIGS. 4A and 4B show portions of a delivery needle and an
insertion needle in, respectively, an insertion position and a
delivery position.
[0033] FIG. 5 is a diagrammatic view of a delivery assembly
according to an embodiment.
[0034] FIG. 6 shows a portion of a needle array, including a
reservoir, according to an embodiment.
[0035] FIG. 7 shows elements of a delivery assembly according to
another embodiment.
[0036] FIG. 8 is a diagrammatic view of a delivery assembly
according to a further embodiment.
[0037] FIG. 9 shows diagrammatically a portion of a tumor
illustrating principles of the invention.
[0038] FIG. 10 is a diagram of a data processing system according
to an embodiment.
DETAILED DESCRIPTION
[0039] The present invention is directed in certain embodiments as
described herein to devices and methods for delivery of fluids to
solid tissues, and in particular embodiments, to solid tumors. The
herein described embodiments relate in part to certain surprising
and heretofore unrecognized advantages, disclosed in greater detail
below, that derive from exquisite control of the location, amount
and time of fluid delivery to solid tissue. These and related
embodiments feature the precise positioning of delivery needle
outlet apertures, including positioning of spatially defined
multiple-needle arrays and/or of needles having multiple outlet
apertures at defined locations, and further including the use of
fluidics configurations that provide extremely fine control over
fluid delivery events. The invention provides improved accuracy and
versatility to screening therapeutic compounds such as anti-cancer
agents for use in treating solid tumors, and permits early
exclusion from a screening program or a therapeutic regimen of
candidate drugs to which tumor cells may be resistant.
[0040] Accordingly, for example, certain embodiments contemplate
direct drug delivery to a solid tissue at low flow rates with low
shear forces that eliminate or reduce mechanochemical damage to
tissues while permitting precisely targeted therapeutic agent
delivery to defined focal sites. These and related embodiments
permit advantageous and selective delivery of a therapeutic agent
to a solid tissue in vivo in a therapeutically effective amount,
while in further related embodiments the agent is undetectable
outside the solid tissue or is present at less than a minimal dose.
Hence, problems (e.g., toxicity, detrimental side-effects, etc.)
associated with administering excessively high systemic
concentrations in order to obtain a therapeutically effective
concentration in a desired solid tissue are overcome by the
presently disclosed embodiments.
[0041] Additionally, certain embodiments contemplate direct
delivery of multiple drugs, candidate drugs, imaging agents,
positional markers, indicators of efficacy and appropriate control
compositions to a plurality of spatially defined locations along
parallel axes in a solid tissue, such as a solid tumor, followed,
after a desired time interval, by excision of the treated tissue
and evaluation or analysis of the tissue for effects of the
treatments. Indicators of efficacy may be, for example, detectable
indicator compounds, nanoparticles, nanostructures or other
compositions that comprise a reporter molecule which provides a
detectable signal indicating the physiological status of a cell,
such as a vital dye (e.g., Trypan blue), a colorimetric pH
indicator, a fluorescent compound that may exhibit distinct
fluorescence as a function of any of a number of cellular
physiological parameters (e.g., pH, intracellular Ca.sup.2+ or
other physiologically relevant ion concentration, mitochondrial
membrane potential, plasma membrane potential, etc., see Haugland,
The Handbook: A Guide to Fluorescent Probes and Labeling
Technologies (10.sup.th Ed.) 2005, Invitrogen Corp., Carlsbad,
Calif.), an enzyme substrate, a specific oligonucleotide probe, a
reporter gene, or the like. Control compositions may be, for
example, negative controls that have been previously demonstrated
to cause no statistically significant alteration of physiological
state, such as sham injection, saline, DMSO or other vehicle or
buffer control, inactive enantiomers, scrambled peptides or
nucleotides, etc.; and positive controls that have been previously
demonstrated to cause a statistically significant alteration of
physiological state, such as an FDA-approved therapeutic
compound.
[0042] Typically and in certain preferred embodiments, the excised
tissue may be cut into a plurality of serial histological sections
along parallel planes that are substantially normal (e.g.,
perpendicular or deviating from perpendicular by as much as 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35 or more
degrees) to the parallel axes, for analysis by any of a number of
known histological, histochemical, immunohistological,
histopathologic, microscopic (including morphometric analysis
and/or three-dimensional reconstruction), cytological, biochemical,
pharmacological, molecular biological, immunochemical, imaging or
other analytical techniques, which techniques are known to persons
skilled in the relevant art. See, e.g., Bancroft and Gamble, Theory
and Practice of Histological Techniques (6.sup.th Ed.), 2007
Churchill Livingstone, Oxford, UK; Kiernan, Histological and
Histochemical Methods: Theory and Practice, 2001 Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, N.Y.; M.A. Hayat (Ed.),
Cancer Imaging--Vols. 1 and 2, 2007 Academic Press, N.Y. Imaging
may be performed before, during or after dispenser needles are
inserted into the solid tissue. Positional markers are known and
include, as non-limiting examples, metal or plastic clips,
fluorescent quantum dots, India ink, metal or plastic beads, dyes,
stains, tumor paint (Veiseh et al., 2007 Canc. Res. 67:6882) or
other positional markers, and may be introduced at desired
positions. Markers may include any subsequently locatable source of
a detectable signal, which may be a visible, optical, colorimetric,
dye, enzymatic, GCMS tag, avidin, biotin, radiological (including
radioactive radiolabel and radio-opaque), fluorescent or other
detectable signal.
[0043] A detectable marker thus may comprises a unique and readily
identifiable gas chromatography/mass spectrometry (GCMS) tag
molecule.
[0044] Numerous such GCMS tag molecules are known to the art and
may be selected for use alone or in combination as detectable
identifier moieties. By way of illustration and not limitation,
various different combinations of one, two or more such GCMS tags
may be added to individual reservoirs of the device described
herein in a manner that permits the contents of each reservoir to
be identified on the basis of a unique GCMS "signature", thereby
permitting any sample that is subsequently recovered from an
injection region to be traced back to its needle of origin for
identification purposes. Examples of GCMS tags include
.alpha.,.alpha.,.alpha.-trifluorotoluene, .alpha.-methylstyrene,
o-anisidine, any of a number of distinct cocaine analogues or other
GCMS tag compounds having readily identifiable GCMS signatures
under defined conditions, for instance, as are available from SPEX
CertiPrep Inc. (Metuchen, N.J.) or from SigmaAldrich (St. Louis,
Mo.), including Supelco.RTM. products described in the Supelco.RTM.
2005 gas chromatography catalog and available from
SigmaAldrich.
[0045] Through the use of the device described herein, which
includes configuration (e.g., by placing at least one positional
marker in one or more known locations) of the multiple needles in a
manner that permits ready identification of the effects at a
particular location, if any, of the contents released from a
particular needle at the tissue location these and related
embodiments thus contemplate methods of simultaneously comparing
the relative therapeutic efficacies and/or toxicities of a large
number of candidate therapeutic agents. Such applications may find
uses in methods of drug screening and drug discovery, such as in
preclinical animal models to identify and functionally characterize
potential new therapeutics. For instance, a plurality of siRNAs may
be administered intratumorally and their relative abilities to
knock down expression of a desired target gene may be compared.
Other similar embodiments may find uses in clinical contexts, for
example, to "deselect", or eliminate from consideration, known
therapeutic agents that have no effect in a particular tumor,
thereby advantageously advancing the therapeutic management of a
patient by avoiding the loss of time and the undesirable
side-effects that may be associated with administering an
ineffectual treatment regimen.
[0046] The present invention provides compositions and methods that
are useful for the classification and/or stratification of a
subject or patient population, including for use in drug discovery
and in pharmacogenomics. In these and related embodiments,
correlation of one or more indicia of an altered physiological
state with a position at which a given candidate agent has been
introduced in a solid tumor may be used to gauge the subject's
responsiveness to, or the potential efficacy of, a particular
therapeutic treatment; related embodiments contemplate this
approach for "deselection", or elimination from consideration as
potential therapies, of candidate agents in which no evidence of an
altered physiological state is detected at a site of introducing in
the tumor.
[0047] As described herein, determination of levels of at least one
indicator of altered physiologic state may also be used to stratify
a patient population for eligibility to participate in a clinical
trial. These and related embodiments are contemplated as usefully
providing advantages associated with evaluation of candidate
therapeutic compounds at an earlier stage of development than is
currently the case. For instance, it is not currently standard
clinical trial practice to establish biomarker parameters (which
may be the basis for exclusion of subjects) prior to Phase Ill
studies, whereas the embodiments described herein may provide
useful results even in the absence of established biomarker
criteria, for example, at Phase II. Accordingly it is envisioned
that through the practice of certain presently disclosed
embodiments, relevant information on the properties of a candidate
agent may be obtained earlier in a solid tumor oncology drug
development program than has previously been the case, including in
a manner which may time-efficiently and cost-effectively permit
elimination from a clinical trial of subjects for whom no response
or benefit can be expected based on a nonresponder result for a
particular candidate agent.
[0048] For example, stratification of a patient population
according to levels of at least one indicator of altered
physiologic state, determined as described herein, may provide a
useful marker with which to correlate the efficacy of any candidate
therapeutic agent being used in cancer subjects, and/or to classify
subjects as responders, nonresponders or possible responders.
[0049] Referring first to FIG. 1, a needle array assembly 100 is
shown, including a plurality of needles 112, a plurality of
reservoirs 114, a plurality of delivery actuators such as, in the
present example, plungers 116, and a controller 102. Each of the
plurality of needles 112 is fixed in position relative to the
others of the plurality of needles, and the plungers are likewise
operatively coupled so as to be fixed in position and
simultaneously actuable. Each of the plurality of needles 112 is in
fluid communication with a respective one of the plurality of
reservoirs 114, and each of the plurality of plungers includes a
first end positioned in a respective one of the plurality of
reservoirs 114. The controller 102 is operatively coupled to second
ends of each of the plurality of plungers 116. The controller is
configured to control actuation of the plungers within the
reservoir with respect to speed, distance, and direction of
movement.
[0050] Movement of the plurality of plungers 116 in a first
direction creates a negative pressure in the respective reservoirs
114, drawing a therapeutic agent or other fluid into the reservoirs
via the respective needle 112, thereby charging the reservoirs.
Each reservoir 114 can be charged with a different agent, or some
or all of the reservoirs can be charged with a common agent.
Movement of the plurality of plungers 116 in a second direction
creates a positive pressure, or overpressure, in the respective
reservoirs 114, forcing the contents of the reservoirs out via the
respective needles 112.
[0051] In this configuration, a relatively small amount of a
plurality of therapeutic agents can be simultaneously delivered
directly to a region of solid tissue 106 for evaluation and
analysis. In some embodiments, the amount of a therapeutic agent
delivered to the tissue is less than 1 .mu.L per needle. The
evaluation of the tissue 106 and the efficacy of the different
therapeutic agents delivered thereto can be used, for example, to
screen potential therapeutic agents for subsequent clinical trials
or to make patient-specific treatment decisions based on the
relative efficacy of the therapeutic agents in the tissue 106.
[0052] According to various embodiments, any number of needles can
be used. For example, as few as one, two, or three needles can be
used, and according to some embodiments, more than one thousand
needles can be used. According to an embodiment, each of the
needles includes a plurality of ports or apertures arranged along
the length of the needle.
[0053] Turning now to FIGS. 2A-2D, various configurations of
needles 120 are shown. FIG. 2A shows a delivery needle 120a
including a plurality of ports 122 in pairs on opposite sides of
the needle, the pairs being evenly spaced along its length. Each
pair is rotated 90 degrees with respect to adjacent pairs of ports
along the length of the needle 120a. When fluid in a reservoir in
fluid communication with the needle 120a is subjected to an
overpressure, it is forced from the needle via the plurality of
apertures 122. Because the reservoir holding the fluid is to the
right of the needle 120a, as viewed in the figures, an overpressure
in the reservoir will result in the largest volume of fluid being
forced from the right-most ports 122, such that the fluid will be
delivered in a progressively diminishing volume along its length
toward the tip-end 124. The relative volume of fluid distributed
from each of the plurality of ports 122 along the needle 120a may
be influenced by a number of factors including, for example,
viscosity of the fluid, the size and concentration of solids
suspended therein, the density, permeability, and wettability of
tissue in which the needle is positioned, the degree of
overpressure, the size of the ports, etc.
[0054] FIG. 2B shows a delivery needle 120b according to another
embodiment, in which ports 122 are largest near the tip-end 124 of
the needle 120b, and the relative size of each of the plurality of
ports is inversely related to a distance of the respective port
from the tip-end of the needle. Thus, while an overpressure of
fluid in the needle will be greatest at the right-most port 122,
that will also be the smallest port, and, conversely, while the
overpressure will be lowest at the left-most port 122, that port
will also be the largest. By appropriate sizing of each of the
ports 122, the needle 120b can be configured to deliver a
substantially equal volume of fluid at any given location along its
axis, or alternatively, the needle can be configured to deliver
fluid according to any selected distribution profile along its
axis, by appropriate selection of the size of the respective ports
122. The size of the apertures can vary along the. length of the
needle from about 0.01 mm or less to about 0.25 mm or more.
[0055] FIG. 2C shows a delivery needle 120c according to an
embodiment in which a distribution density of the plurality of
ports 122 is inversely related to a distance of the respective port
from the tip-end 124 of the needle 120c. In other words, the ports
122 closest to the tip-end 124 of the needle 120c are the most
closely spaced, while the spacing between the ports grows
increasingly greater as the distance from the tip-end increases.
Accordingly, when fluid in the associated reservoir is subjected to
an overpressure, the volume of fluid per port 122 will be greatest
at the right-most port, but a lower volume of fluid per port will
be offset toward the left by the progressively closer spacing of
the ports. Thus, the overall distribution of fluid along the length
of the needle 120c can be made to be substantially consistent by
distributing the ports 122 as described above, or can be made to
conform to another selected distribution profile by appropriate
selection of the distribution density of the ports along the
needle.
[0056] Turning to FIG. 2D, a delivery needle 120d is shown
according to another embodiment. Ports 126 of the needle 120d are
formed in a spiral pattern, with each port rotated 90.degree. with
respect to adjacent ports. In the embodiment shown in FIG. 2D, the
ports 126 are formed by wire-electrode electrical discharge
machining (wire EDM). In cutting the ports 126, the depth and the
length of each cut can be selected to control the port size, while
the pitch of the spiral can be selected to control the distribution
density. Thus, ports 126 configured as shown in FIG. 2D can be
differentially sized or spaced as described with reference to FIGS.
2B and 2C.
[0057] In addition to wire EDM, the ports 122, 126 of the needles
120 can be formed by any appropriate method, including, for
example, laser cutting, waterjet cutting, chemical etching,
mechanical drilling or grinding, etc.
[0058] The tip-ends 124 of the needles 120 are shown as being
closed and pointed. According to some research, "pencil point"
needles, such as, for example, Sprotte and Whitacre needles, may be
less damaging to biological tissue than bevel-tipped needles.
Additionally, fluids injected into tissue using pencil point
side-port needles tend to remain in the tissue rather than leaking
from the tissue via a channel formed by the needle. Such
considerations are explored in more detail in U.S. Patent
Application No. 2004/0191225--see also U.S. Pat. No.
5,848,996--which are incorporated herein by reference in their
entireties. Nevertheless, the scope of the invention is not limited
to pencil point needles. Bevel-tipped and blunt-tipped needles can
also be employed according to various embodiments. In particular,
the inventors have conducted tests using prototypes with
blunt-tipped needles, which performed satisfactorily.
[0059] FIG. 3 shows a solid-core delivery needle 130 having a
plurality of annular grooves 132. The needle 130 is a "passive
delivery" device, meaning that a therapeutic agent is not delivered
under pressure from a reservoir but instead is carried into the
tissue in the grooves 132. Other passive delivery-type needles
include, for example, needles with micro pits over their surfaces,
needles coated with nanowire, and needles made from porous
materials. Such a needle is immersed in a liquid agent for
sufficient time to charge, and is then inserted into the target
tissue. In the case of the needle 130 of FIG. 3, the needle can be
charged by being briefly dipped into the agent. A porous needle
will carry more of the agent, but may require more time to charge,
and may likewise need to be left in place in the tissue for a
longer period to deliver its charge.
[0060] Embodiments are primarily described herein as using active
delivery needles, i.e., needles that actively force fluid into the
surrounding tissue. However, passive delivery needles such as those
described above can also be employed, according to the design
parameters of a given application.
[0061] FIGS. 4A and 4B show a portion of an inserter needle 140 and
a portion of a delivery needle 120 similar to those described with
reference to FIGS. 2A-2D. In FIG. 4A, the inserter needle 140 is
positioned such that the tip-end 124 of the delivery needle 120
extends slightly beyond an end of the inserter needle 140. In this
configuration, the needle 120 and inserter needle 140 can be
inserted through the skin of a subject, such as a patient or test
model. The combination of the needle 120 and inserter needle 1406
are configured to have sufficient stiffness to penetrate the skin
without bending, and the tapered point of the tip-end 124 assists
in the penetration. Additionally, the inserter needle 140 covers
the ports 122 of the needle 120 and prevents contamination of the
contents of the needle by non-target tissue, and vice-versa. When
the tip-end 124 of the needle has penetrated to within a small
distance of the target tissue, the inserter needle 140 is held in
position while insertion of the needle 120 continues until it is
correctly positioned in the target tissue. The insertion distance
of the inserter needle 140 can be selected such that the needle 120
is correctly positioned once all of the ports 122 are clear of the
inserter needle, as shown in FIG. 2B. In this way, the needle 120
can be provided with maximum protection and support, and the
likelihood of contamination can be minimized.
[0062] According to an alternate embodiment, a stylette is
positioned in the inserter needle to stiffen the needle and prevent
collection of a tissue plug during insertion. Once the inserter
needle 140 is positioned, the stylette is removed and the delivery
needle 120 is inserted.
[0063] FIG. 5 is a diagrammatic view of a delivery assembly 150
according to another embodiment. The delivery assembly 150 includes
a needle array 152, an inserter assembly 154, an actuator assembly
156, a driver assembly 158, a control assembly 160, and a frame
162. The frame 162 provides a substantially rigid structure to
which other elements of the assembly 150 are coupled.
[0064] The needle array 152 comprises a plurality of needle
cylinders 166 and a needle block 168. In the embodiment shown, the
needle block 168 is integral with the frame 162. Each of the
plurality of needle cylinders 166 is coupled, at a first end 170,
in a respective needle aperture 174 extending in the needle block
168, and comprises a lumen 176, having, in the illustrated
embodiment, a nominal diameter of 0.15 mm, extending substantially
the entire length of the needle cylinder 166. Each needle cylinder
166 includes a reservoir 178 in a region toward the first end 170,
a needle 120 in a region toward a second end 180, and a tip-end 124
at the second end 180 of the needle cylinder 166. In the embodiment
shown, the tip-end 124 is tapered to a point.
[0065] Each delivery needle 120 is defined by a plurality of ports
122 distributed along its length. The length of each of the
plurality of needle cylinders 166 and of the respective needles 120
varies according to the embodiment. In one embodiment, each needle
cylinder 166 is longer than 15 cm, while according to other
embodiments the needle cylinders are each longer than 10 cm,
between 5 cm and 10 cm, and as short as 2 cm, respectively.
Likewise, according to various embodiments, each of the plurality
of delivery needles 120, defined by the portion of the respective
needle cylinder 166 along which the ports 122 are spaced, is longer
than 1 cm, longer than 2 cm, longer than 4 cm, and longer than 8
cm.
[0066] The inserter assembly 154 comprises a plurality of inserter
needles 140 coupled to an inserter block 192 in respective inserter
apertures 190 extending therein in a configuration that corresponds
to the arrangement of the needle cylinders 166 in the needle block
168, such that each of the plurality of needle cylinders 166 can be
positioned within a respective one of the plurality of inserter
needles 140 as shown in FIG. 5. The inserter assembly 154 is
axially slidable over the needle cylinders 166 between a first
position, in which only the tip-ends 124 of each of the needle
cylinders 166 extend from respective ones of the plurality of
inserter needles 140, to a second position, in which the second
ends 180 of each of the needle cylinders 166 extends from the
respective inserter needle 140 a distance sufficient to clear all
of the ports 122 of the respective delivery needle 120.
[0067] According to an embodiment, a spacer is provided, configured
to be positioned between the inserter block 192 and the needle
block 168, sized such that when the inserter block and the needle
block are both engaged with the spacer, the inserter block is
maintained in the first position. Removal of the spacer permits
movement of the inserter block 192 and the needle block 168
relative to each other, to permit placement of the inserter block
into the second position, relative to the needle block.
[0068] The actuator assembly 156 comprises a plurality of plungers
200 coupled at respective first ends 204 to a plunger block 206 in
a configuration that corresponds to the arrangement of the needle
cylinders 166 and the inserter needles 140 such that a second end
208 of each of the plurality of plungers 200 can be positioned
within the reservoir 178 of a respective one of the plurality of
the needle cylinders 166 as shown. An O-ring 210 is provided at the
second end 208 of each of the plurality of plungers 200 to
sealingly engage the wall of the respective lumen 176. The actuator
assembly 156 also comprises an actuator 212 coupled to an actuator
block 214, which in turn is rigidly coupled to the plunger block
206. In the embodiment shown, the actuator 212 comprises a
micrometer device 220 having a thimble 222, a barrel 224, and a
spindle 228 such as are well known in the art. The barrel 224 is
rigidly coupled to the frame 162 while the spindle 228 is rotatably
coupled to the actuator block 568 so as to control translational
movement of the actuator block relative to the frame 162. The
micrometer device 568 is calibrated in 0.01 mm increments, with a
spindle travel of 0.5 mm per rotation of the thimble 222 and a
maximum stroke of 15 mm. Thus, each complete rotation of the
thimble moves each of the plurality of plungers 0.5 mm within the
lumen 178 of the respective needle cylinder 166 and displaces about
0.0001 cm.sup.3 of volume, or 0.1 nL per revolution. Thus, given a
maximum stroke of 15 mm, the maximum dispensing capacity of each of
the plurality of needles 120 is about 3 nL.
[0069] The driver assembly 158 comprises a stepper motor 230 such
as is well known in the art, and that includes a motor casing 232,
a motor shaft 234 coupled to a rotor of the motor 230, and other
elements such as are well known in the art. The motor casing 232 is
rigidly coupled to the frame 162, and the motor shaft 234 is
slidably coupled to the thimble 222 of the micrometer device 568
while being rotationally locked therewith, such as via a spline
coupling, for example. Accordingly, rotational force from the motor
shaft 234 is transmitted to the thimble 222, while axial movement
of the thimble is not limited by the motor shaft. Such couplings
are well known in the mechanical arts. The stepper motor 230 of the
illustrated embodiment is configured to divide each rotation into
125 steps. Thus, each incremental rotational step of the motor 230
rotates the thimble about 3.degree., displacing a volume of about
0.8 pL per reservoir 178.
[0070] The controller assembly 160 includes a controller 240 and a
control cable 242 that extends from the controller to the stepper
motor 230. Signals for controlling direction, speed, and degree of
rotation of the motor shaft 234 are transmitted from the controller
240 to the stepper motor 230 via the control cable 242 in a manner
that is well known in the field to which such motors belong.
According to an embodiment, the controller is programmable. A user
can program the controller to control a speed of delivery of a
fluid from the delivery needles 120 by selecting the speed of
rotation, and a volume of fluid delivered by selecting the number
of partial and complete rotations of the rotor. According to
another embodiment, the controller is manually operated, such that
a user controls a rate and direction of rotation of the motor 230
in real time. According to a third embodiment, the driver and
controller assemblies are omitted, and a user controls fluid
delivery by manually rotating the thimble 222 of the actuator
assembly 212.
[0071] Charging the reservoirs 178 can be accomplished in a number
of ways. For example, a charging vessel can be provided that
includes a plurality of cups or compartments in an arrangement that
corresponds to the arrangement of the needle cylinders 166. The
user first places a selected fluidic agent or combination of agents
in each of the cups. The delivery assembly 150 of FIG. 5 is
positioned with the needle cylinders pointing downward as shown in
the drawing, and the spindle 228 of the actuator 212 fully
extended. The frame 162 is lowered until the needles 120 are fully
immersed in the fluids in the respective cups. The motor 230 is
then controlled to rotate in the reverse direction, drawing the
spindle 228 inward and pulling the plungers 200 upward. This in
turn creates a negative pressure in the reservoirs 178 relative to
ambient, drawing the fluids into the needle cylinders 166 via the
needle ports 122. When the reservoirs are sufficiently charged,
rotation of the rotor is halted and the needle array 152 is
withdrawn from the charging vessel.
[0072] In order to deliver the charge, according to one embodiment,
each of the needle cylinders 166 of the needle array 152 is
positioned in a respective one of the inserter needles 140 of the
inserter assembly 154 so that the tip-ends 178 of the needles 120
protrude from the inserter needles 140, substantially as described
with reference to FIG. 4A. The delivery assembly 150 is then
positioned in axial alignment with a target tissue region of a
subject and translated axially so that the tip-ends of the needles
120 penetrate the subject's skin. Axial translation of the delivery
assembly 150 continues until the tip-ends 124 of the needle
cylinders 166 have penetrated to within a selected distance of the
target tissue region. The inserter assembly 154 is then held in
position while the frame 162 and the elements coupled thereto
continue to move axially, such that the needles 120 extend into the
target tissue region. When the needles 120 are correctly
positioned, movement of the delivery assembly 150 is halted and the
frame 162 is held in position relative to the subject. The stepper
motor 230 is then controlled to rotate the thimble 222 in the
forward direction so as to cause the spindle 228 to extend, driving
the plungers 200 into the needle cylinders 166 and creating an
overpressure in the respective reservoirs 178, thereby forcing
fluid from the reservoirs to the target tissue region via the ports
122 of the delivery needles 120.
[0073] Delivery can be performed in a few seconds, or it can be
extended over minutes or hours under a relatively low overpressure
to promote complete absorption of the fluid into the surrounding
tissue. According to the embodiment described with reference to
FIG. 5, the stepper motor 230 can be controlled to rotate the rotor
fast enough to depress the plungers 200 the full 15 mm in less than
one second, or slow enough that a single rotation can take many
hours.
[0074] FIG. 6 shows a portion of a needle array 250 according to
another embodiment. A portion of a needle cylinder 166 is shown,
together with a portion of a needle block 252. The first end 170 of
the needle cylinder 166 is coupled to a first portion 254 of an
aperture 256 extending in the needle block 252. The aperture 256
includes the first portion 254, sized to receive the needle
cylinder 166, and a second portion 258 having an increased
diameter. In the embodiment shown, the diameter of the second
portion 258 has a diameter of 0.75 mm. The second portion 258
defines a reservoir that is in fluid communication with the needle
cylinder 166. The second end of a plunger 260 is positioned in the
second portion 258, with an O-ring 262 sealingly engaged
therein.
[0075] In the arrangement described with reference to the delivery
assembly 150 of FIG. 5, the second end of each of the plurality of
plungers 200 is positioned in a respective one of the needle
cylinders 166, and the reservoirs 178 are comprised by the needle
cylinders. Thus, axial movement of one of the plurality of plungers
200 within the lumen 176 of a respective needle cylinder 166
displaces a volume equal to a transverse cross-sectional area of
the lumen, multiplied by the distance of travel of the plunger. In
contrast, because of the diameter of the second portion 258 of the
aperture 256 of FIG. 6, relative to the diameter of the lumen 176,
axial movement of the plunger 260 displaces a volume that is
greater than the volume displaced by a plunger 200 of FIG. 5 by a
factor of 25, for a given distance of travel. Conversely, to
displace an equal volume of the reservoir 178, a plunger 200 of
FIG. 5 must travel 25 times as far as the plunger 260 in the second
portion 258 of the aperture. Thus, given otherwise identical
elements, the delivery assembly 150 of FIG. 5 is capable of
accurately metering delivery of smaller quantities of fluid than a
similar assembly having reservoirs and plungers configured as
described with reference to FIG. 6, while the latter is capable of
delivering larger quantities of fluid for a given plunger stroke
length; up to 75 nL over a stroke of 15 mm vs. 3 nL for the
embodiment of FIG. 5. Of course, the values given are exemplary;
one of ordinary skill will recognize that the needle sizes as well
as reservoir sizes can be selected to accommodate particular
requirements.
[0076] Turning now to FIG. 7, elements of a delivery assembly 270
are shown according to another embodiment. A needle block 272
includes a large plurality of needle apertures 274 extending
therethrough, arranged in a closely spaced array. Needle cylinders
166 are provided separately, in various assortments of lengths and
numbers, sizes, and spacings of ports.
[0077] In use, a user selects a number of needles to be used for a
particular procedure, and selects the particular needle cylinders
166, placing each in a respective one of the plurality of apertures
274 of the needle block, in an arrangement that is selected for the
particular procedure. The user may require only a small number of
needles, such as one to five, for example, or may require hundreds
or thousands of needles. Furthermore, the needle cylinders 166 can
be of varying lengths and configurations. The user selects the
arrangement of the needle cylinders 166 in the needle block 272,
and their respective lengths and configurations, at least in part
according to factors such as the size, shape, and position of a
target tissue region in a subject's body, the desired distribution
density of fluid in the target tissue region, the permeability of
the target tissue, etc.
[0078] The needle cylinders 166 can be affixed in the apertures 274
by any appropriate means, including, for example, soldering,
brazing, and by adhesive. Alternatively, the needle cylinders 166
can be sized and configured to be fixed in place by an interference
fit, such that, for example, the first end of each needle cylinder
has a fractionally increased outer diameter. The user drops each
needle cylinder into a respective aperture, tip-end first, then
pulls the needle cylinder into the aperture from the other side of
the needle block until the first end is firmly engaged in the
aperture. A plunger block and an inserter block are also provided,
with respective pluralities of apertures in arrays corresponding to
the array of needle apertures of the needle block. The user loads
plungers 200 and inserter needles 140 concurrently with the needle
cylinders 166 for operation in a delivery assembly similar to that
described with reference to FIG. 5. Provided the method used to
attach the needle cylinders 166 in place can be reversed, the
needle block 272 can be reused repeatedly for different
procedures.
[0079] The needle block 272 shown in FIG. 7 is about 5 cm in
diameter, 1 cm in thickness, and has approximately 1,600 apertures,
spaced about 1 mm apart. According to other embodiments, the needle
block can be any appropriate shape and size, from as small as 1 or
2 centimeters across to as large as ten or more centimeters across,
and can have any number of apertures, from ten or fewer to several
thousand. According to various embodiments, the needle block is
provided, for example, with 200, 400, and 800 apertures. The large
number of apertures provides significant freedom to a user to
control spacing between needles as well as the particular pattern
of the array. The needle block 272 is shown with a hexagonal grid
array of apertures. The apertures can also be arranged in other
grid configurations, such as, for example, rectangular and
quincunx. The needle cylinders shown are about 5 cm in length, but
this too is merely exemplary.
[0080] The delivery actuators of previous embodiments have been
described as plungers. However, any suitable actuator can be used
to control an amount of therapeutic agent delivered from the
reservoirs into the needle. For example, fluid pressure such as by
compressed air or pressurized liquid can be used to control an
amount of therapeutic agent delivered to a region of biological
tissue via the reservoirs and needles.
[0081] Referring now to FIG. 8, a delivery assembly 300 is shown,
according to another embodiment. The delivery assembly 300 includes
a plurality of needle cylinders 302 comprising respective
reservoirs 178 and needles 120. Fluid couplings 312 place the
needle cylinders 302 in fluid communication with a manifold 304. A
fluid pressure source 306 and a fluid vacuum source 308 can each be
placed in fluid communication with the manifold 304 by operation of
a valve 310.
[0082] According to the embodiment of FIG. 8, the needle cylinders
302 are not fixed with respect to each other, but can be
individually emplaced, in a target tissue region, for example. The
reservoirs are first charged, by placing the delivery needles 120
in a selected fluid, e.g., a therapeutic agent or respective
therapeutic agent, and the fluid vacuum source is placed in fluid
communication with the manifold, drawing a negative pressure into
the reservoirs and drawing the agent into the needles. The user
then positions the needles 120 in the target tissue region. When
they are all in place, the manifold 304 is pressurized, forcing
fluid from the reservoirs of each of the needle cylinders 166 via
the ports 122 of the respective delivery needles. While FIG. 8
shows a simple fluid circuit, it will be understood that in
practice such a circuit could include any of valves, pressure
regulator, peristaltic pump, microfluidic pump, vacuum accumulator,
compressor, controller, etc., all of which are well known in the
art, and within the abilities of one of ordinary skill to select
and configure for a given application.
[0083] According to an embodiment, solid tissue into which a
plurality of therapeutic agents have been delivered is subsequently
resected from the subject and evaluated. For example, in a case
where the target tissue is a cancerous tumor, the plurality of
agents injected therein can include some agents whose efficacy or
effect on such tumors is under investigation. By injecting the
various agents in vivo then waiting a selected period before
removing the tumor, the effect of the agents on the tumor in situ
can be investigated. This preserves the tumor microenvironment and
distinguishes this method from current ex vivo or in vitro
therapeutics evaluation methods. Assuming that the needles used are
configured to deliver a substantially equal amount of fluid at any
given location along their length, as described above with
reference to FIGS. 2B-2D, the agent delivered by each of the
needles is evenly distributed to the surrounding tissue along the
delivery axis on which the respective needle 120 was positioned
during the delivery of the agent to the tumor 320. Over time, each
agent permeates outward from its delivery axis to a greater or
lesser degree, depending on factors such as, for example, the
density of the surrounding tissue, the viscosity and composition of
the agent, the wettability of the tissue by the respective agent,
etc. Typically, the portions of the tissue into which the agents
spread are approximately column-shaped regions coaxial with the
respective delivery axes.
[0084] According to various embodiments, a region of tissue is left
in place for some period of time before being resected. For
example, 48-72 hours following delivery is thought to be generally
sufficient for a tumor to exhibit a detectable response. In other
cases, the wait period may be hours, days, or weeks. According to
some embodiments, the tissue region is imaged using known methods
to precisely locate the target region of tissue prior to insertion
of the needles. The region may be imaged repeatedly before and
after delivery of the plurality of agents to the region of
tissue.
[0085] According to other embodiments, a plurality of agents are
delivered to a portion of tissue via respective ones of a plurality
of needles of a needle array after the portion of tissue is
resected.
[0086] Referring now to FIG. 9, a portion of a tumor 320 is shown,
following an injection procedure and subsequent resection. The
tumor 320 has been sectioned into a plurality of slices 322 along
planes that lie substantially normal to the delivery axes.
Column-shaped delivery regions 324 define the regions of permeation
of the respective agents, and extend perpendicular to the planes of
the sections 322.
[0087] Many of the regions 324 may not be easily detectable to a
user, so generally at least two readily detectable position markers
324a, 324b are among the agents injected, at widely separated
locations. The user can then overlay a template on which the
locations of each of the delivery axes is marked, aligning the
indicated marker positions of the template with the detectable
position markers 324a, 324b of a given section 322, thereby
locating the remaining delivery regions 324. The position markers
324a, 324b can be any composition that is detectable by a user.
Various exemplary position markers are described in detail
elsewhere in this disclosure. According to an embodiment, the
position markers are selected to resist permeation and diffusion
into the surrounding tissue and to remain concentrated in a narrow
column, as shown for example at 324a, so as to be detectable for an
extended period after the injection procedure, and to provide an
accurate guide for positioning the template.
[0088] In addition to position markers, control agents may also be
among the agents injected. For example, a negative control can
comprise a substance used as a vehicle in others of the agents, and
a positive control can comprise a compound of most or all of the
agents delivered individually at other delivery axes.
[0089] Following sectioning of the tumor 320, a user conducts
selected assays on delivery regions 324 of various sections 322 of
the tumor 320, as described in more detail later. One benefit of
the devices and methods disclosed herein is that, in addition to
evaluating the efficacy of a given agent on the tumor, the efficacy
of agents at various delivery regions 324 can be evaluated and
compared. Additionally, the effect of a given agent on various
parts of the tumor can be evaluated, both vertically and
horizontally. By comparing the effect of an agent in a delivery
region 324c at section 322a, for example, with its effect in the
same region 324c at sections 322b and 322c, the effect of that
agent on different tissue compositions that may occur vertically
can be differentiated. Similarly, the same agent can be delivered
at several delivery axes in the array, e.g., 324c and 324d, and the
relative effects at those locations in a given section 322 can then
be compared, providing horizontal differentiation. As is well known
in the art, biological tissue is rarely homogeneous over even
relatively small distances. A given agent might have substantially
no effect on some tissue structures of a tumor, but might, on the
other hand, be extremely effective on others. Such differential
effects can be detected and evaluated as described above.
[0090] Another valuable aspect that can be evaluated is the effect
of multiple agents in regions where they interact within the
tissue. Delivery regions 324e and 324f are spaced more closely
together than the others, resulting in the respective agents
interacting in a region 324ef where the respective delivery regions
overlap.
[0091] As discussed in the background section of this disclosure,
clinical trials for cancer related therapeutics are incredibly
expensive and time consuming. It is therefore very important to
effectively screen for agents that have relatively greater
potential as early in the process as possible. Agents subjected to
such screening are sometimes referred to as candidate effective
agents. One screening method involves placing each candidate agent
in a respective Petri dish with a growth medium. A cancerous tumor
is reduced to a homogeneous slurry and is distributed among the
Petri dishes and incubated.
[0092] The dishes are later evaluated for indications of cell
growth. Agents that appear to have impeded growth of cancer cells
may then be advanced for further study.
[0093] However, this method is only marginally effective, for
several reasons. First, many cancers are known to be nonviable
outside a live subject, for reasons such as lack of a blood supply,
etc., and fail to grow in vitro under any circumstances. Screening
tests like that described are therefore ineffective with these. In
some cases it is not known that a particular strain falls in this
category prior to conducting the test. The result is that an
expensive and time consuming test is inconclusive, and the tumor
cannot be salvaged for an alternative test. If the tumor is of a
rare strain, it may be some time before another is available for
alternative testing.
[0094] Second, the reduction process can alter the response
characteristics of a tumor. The process involves essentially
pureeing the tumor, which completely destroys any structural
differentiation, and may render the cancer susceptible to some
agents that would have no effect on the same strain in vivo,
resulting in a false positive, even though such agents might be
useless for treating the cancer in patients. The result is that
many such agents are not eliminated until later phases of study,
after much more money and effort have been expended.
[0095] Third, the same reduction process can also produce false
negatives, in which some agents may fail to inhibit cell growth in
vitro, but would be effective in treating the same cancer in vivo.
This results in the premature elimination of some agents that might
otherwise have become effective therapeutic options.
[0096] Many false positives or false negatives are generated in the
current ex vivo or in vitro art because tumor cells are separated
from their microenvironment, e.g., surrounding non-cancerous cells,
blood, hormones, paracrine factors, oxygen tension, cell-cell
communications, and host immune functions, all of which may
influence whether certain therapeutic agents have or do not have
activity in cancer cells.
[0097] Fourth, even where accurate, only the most general
information can be gleaned from such studies because the test
conditions do not remotely resemble the conditions in which the
cancer normally lives and grows, and in which it is treated
therapeutically.
[0098] It is also known to inject a test agent into a tumor prior
to resection from a subject, for subsequent examination. However,
in such tests a single agent is typically injected, so they are not
feasible for early screening, but are usually reserved for agents
that have already demonstrated significant potential. Even in
animal models, it is expensive and time consuming to induce a tumor
and allow it to grow to a practical size, which makes extensive
early screening by this method impractical.
[0099] Finally, even where general efficacy of an agent in treating
a particular cancer type, subtype, variant, strain or the like has
been demonstrated, it is not uncommon for the cancer of a
particular patient to be wholly unresponsive to the agent. The
patient is thus exposed to the often extreme discomfort and
toxicity of the treatment--not to mention the cost--without
significant benefit. Worse, because the agent's ineffectiveness may
not be known for a long period while the treatment is ongoing, the
opportunity to shift to a different treatment that might have been
completely successful may be lost.
[0100] Where a similar idiosyncratic response--or lack
thereof--occurs in a subject of a drug study, the results of the
study can be skewed. To avoid this, it is typically necessary to
resort to larger test groups to minimize the statistical impact of
nonresponding study subjects, which further increases the cost of
such studies.
[0101] The inventors have recognized that the inability in the
known art to accurately position an agent in tissue in vivo,
especially with respect to other agents, and the inability to later
identify the locations of agents in tissue, prevent more extensive
and beneficial use of in vivo injection, and likewise, that if such
accuracy could be achieved, significant benefits in research and
therapy could be realized.
[0102] It has been noted above that the volume of fluid that is
delivered by each delivery needle can be vanishingly small, much
less than would be a minimal dose required to produce a detectable
effect in an adult. Depending on the agent, the effect may
nevertheless be detected on the very small region immediately
surrounding the delivery site. Accordingly, candidate effective
agents can be injected into a tumor, for example, in situ, without
danger of harming the subject. Additionally, a significant number
of different agents can be simultaneously delivered to respective
delivery axes within the tumor.
[0103] The procedures described above can be employed to resolve a
number of the problems and difficulties that contribute to the cost
and delay of developing effective cancer therapies. For example,
because the candidate agents are delivered in vivo, the tumor is
not otherwise disturbed, and so its reaction to each agent will
tend to be indicative of its reaction if exposed to that agent in
therapeutically effective quantities. The incidence of false
positives and false negatives is significantly reduced.
[0104] Second, because relatively large numbers of agents can be
delivered to a tumor without significant danger to the subject, it
is practical to use the procedure to screen large numbers of
candidate agents early in the testing process, perhaps eliminating
those that show the least promise, flagging the most promising
agents for additional study, or prioritizing candidates for further
study.
[0105] Third, again because of the large number of agents that can
be delivered to a tumor, potential study subjects can be screened
for response to particular agents, reducing or eliminating the
number of subjects with idiosyncratic responses.
[0106] Fourth, when employed in a therapeutic setting, a patient
can be tested for response to a large number of treatments and the
most promising can be identified early in the process, thereby
reducing the number of patients who undergo ineffective treatments
and improving the likelihood that a patient will receive the most
effective available treatment.
[0107] According to an embodiment, a plurality of candidate
effective agents are delivered, in vivo, along mutually parallel
axes to a region of solid tissue, substantially as described above.
The region of solid tissue is subsequently resected from a subject
and an effect of each of the plurality of agents on the solid
tissue is evaluated. Based on the evaluation, one or more of the
plurality of agents are prioritized for further investigation.
[0108] According to alternate embodiments, one or more of the
plurality of agents are selected or deselected for further
investigation, based on the evaluation.
[0109] According to another embodiment a plurality of agents that
have been shown to have therapeutic benefits are delivered, in
vivo, along mutually parallel axes to a region of solid tissue. The
region of solid tissue is subsequently resected from a subject and
an effect of each of the plurality of agents on the solid tissue is
evaluated. Based on the evaluation, one or more of the plurality of
agents are prioritized for therapeutic treatment of the subject.
According to alternate embodiments, one or more of the plurality of
agents are selected or deselected for therapeutic treatment of the
subject, based on the evaluation.
[0110] According to another embodiment a plurality of agents that
are currently under investigation for therapeutic efficacy are
delivered, in vivo, along mutually parallel axes to a region of
solid tissue in each of a plurality of subjects. The respective
regions of solid tissue are subsequently resected from each of the
subjects and an effect of each of the plurality of agents on the
respective regions of solid tissue is evaluated. Based on the
evaluation, one or more of the plurality of subjects are selected
as candidates for further study of the therapeutic efficacy of one
or more of the agents. According to alternate embodiments, one or
more of the plurality of subjects are deselected as candidates for
further study of the therapeutic efficacy of one or more of the
agents.
[0111] According to some embodiments, needle arrays are employed to
effectively and broadly deliver therapeutic agents to live and
viable tissue (e.g., solid tissue) in a subject, wherein imaging is
not required and the tissue is not subsequently resected. The
subject may be monitored according to appropriate clinical criteria
for assessing clinical improvement. In these and related
embodiments, it will be appreciated that significantly higher
levels of the therapeutic agent will be achieved within the solid
tissue than would be the case if the agent were delivered
systemically, although detectable amounts of the therapeutic agent
may be subsequently identified outside the solid tissue (e.g., in
the circulation). As a non-limiting example, one such embodiment
contemplates direct intramuscular introduction of a gene therapy
agent for treating muscular dystrophy (e.g., an engineered
therapeutic virus, a therapeutic agent-carrying nanoparticle, etc.)
to one or more skeletal muscle injection sites in a subject,
without the need for imaging, surgery, or histology on biopsy
specimens. Of course, periodic monitoring of the circulation for
leaked therapeutic agent and/or subsequent analysis of a biopsy
specimen, e.g., to assess the effects of the agent on the target
tissue, may also be considered.
[0112] In other embodiments it is contemplated that the target
region in a solid tissue may be imaged using known techniques to
evaluate the effects of the agents. The imaging can be by any
suitable process or method, including, for example, radiographic
imaging, magnetic resonance imaging, positron emission tomogoraphy,
biophotonic imaging, etc. In some embodiments, the target region
may be imaged repeatedly before, during, and after the delivery
process.
[0113] According to the embodiment of FIG. 10, a data processing
system 350 is used to carry out or direct operations, and includes
a processor 354 and a memory 356. The processor 354 communicates
with the memory 356 via an address/data bus 360 and also
communicates with a needle array assembly 362 and a patient
scanning device 364. The patient scanning device 364 is used,
according to an embodiment, to assist in placing the needles of the
needle array assembly 362 in a patient in vivo and for non-invasive
analysis of target tissue regions using imaging techniques, such as
radiographic imaging or nuclear medical assays. The processor 354
can be a commercially available or custom microprocessor,
microcontroller, signal processor or the like. The memory 356 can
include any memory devices and/or storage media containing the
software and data used to implement the functionality circuits and
modules.
[0114] The memory 356 can any of include several categories of
software and data used in the data processing system, such as, for
example, an operating system 366, application programs 368;
input/output device drivers 370; and data 372. The application
programs 368 are illustrative of the programs that implement the
various features of the circuits and modules according to some
embodiments, and the data 372 represents the static and dynamic
data used by the application programs 368, the operating system
366, the input/output device drivers 370 and other software
programs that may reside in the memory 356.
[0115] According to various embodiments, the data processing system
350 may include several modules, including an array controller 376,
a scanner controller 378 and the like. The modules may be
configured as a single module or additional modules otherwise
configured to implement the operations described herein. For
example, the array controller 376 can be configured to control the
needle array assembly 100 of FIG. 1, by controlling the actuators
116, and consequently, the release of therapeutic agents from the
reservoirs 114 via the needles 112. The scanner controller 378 can
be configured to control the patient scanning device 364.
[0116] Certain embodiments described herein relate to introducing
an agent into a solid tissue in a subject, and/or excising all or a
portion of a solid tissue from a subject, and/or obtaining one or
more biological samples from a solid tissue that may be in a
subject, and/or screening one or more subjects for clinical trial
eligibility, and/or any number of other methods that may involve a
subject, which includes a subject or biological source.
[0117] The subject or biological source may be a human or non-human
animal, a transgenic or cloned or tissue-engineered (including
through the use of stem cells) organism, a primary cell culture or
culture adapted cell line including but not limited to genetically
engineered cell lines that may contain chromosomally integrated or
episomal recombinant nucleic acid sequences, immortalized or
immortalizable cell lines, somatic cell hybrid cell lines,
differentiated or differentiatable cell lines, transformed cell
lines and the like. In certain preferred embodiments of the
invention, the subject or biological source may be suspected of
having or being at risk for having a malignant condition, and in
certain preferred embodiments of the invention the subject or
biological source may be known to be free of a risk or presence of
such disease.
[0118] Certain preferred embodiments contemplate a subject or
biological source that is a human subject such as a patient that
has been diagnosed as having or being at risk for developing or
acquiring cancer according to art-accepted clinical diagnostic
criteria, such as those of the U.S. National Cancer Institute
(Bethesda, Md., USA) or as described in DeVita, Hellman, and
Rosenberg's Cancer: Principles and Practice of Oncology (2008,
Lippincott, Williams and Wilkins, Philadelphia/Ovid, New York);
Pizzo and Poplack, Principles and Practice of Pediatric Oncology
(Fourth edition, 2001, Lippincott, Williams and Wilkins,
Philadelphia/Ovid, New York); and Vogelstein and Kinzler, The
Genetic Basis of Human Cancer (Second edition, 2002, McGraw Hill
Professional, New York); certain embodiments contemplate a human
subject that is known to be free of a risk for having, developing
or acquiring cancer by such criteria.
[0119] Certain other embodiments contemplate a non-human subject or
biological source, for example a non-human primate such as a
macaque, chimpanzee, gorilla, vervet, orangutan, baboon or other
non-human primate, including such non-human subjects that may be
known to the art as preclinical models, including preclinical
models for solid tumors and/or other cancers. Certain other
embodiments contemplate a non-human subject that is a mammal, for
example, a mouse, rat, rabbit, pig, sheep, horse, bovine, goat,
gerbil, hamster, guinea pig or other mammal; many such mammals may
be subjects that are known to the art as preclinical models for
certain diseases or disorders, including solid tumors and/or other
cancers (e.g., Talmadge et al., 2007 Am. J. Pathol. 170:793;
Kerbel, 2003 Canc. Biol. Therap. 2 (4 Suppl 1):S134; Man et al.,
2007 Canc. Met. Rev. 26:737; Cespedes et al., 2006 Clin. Transl.
Oncol. 8:318). The range of embodiments is not intended to be so
limited, however, such that there are also contemplated other
embodiments in which the subject or biological source may be a
non-mammalian vertebrate, for example, another higher vertebrate,
or an avian, amphibian or reptilian species, or another subject or
biological source.
[0120] Biological samples may be provided by obtaining a blood
sample, biopsy specimen, tissue explant, organ culture, biological
fluid or any other tissue or cell preparation from a subject or a
biological source. In certain preferred embodiments the biological
sample may be obtained from a solid tissue (e.g., a solid tumor)
using the herein described device, for example, by introducing a
multiple needle device into a solid tissue, thereby placing a
plurality of needles at a plurality of positions in the tissue, and
generating negative pressure at one or a plurality of ports of each
needle of the multiple needle device under conditions and for a
time sufficient to draw into the needles a plurality of biological
samples from the plurality of positions in the tissue.
[0121] Devices and methods disclosed here may find uses according
to certain preferred embodiments for the introduction of agents to,
and/or the withdrawal of biological samples from, a solid tissue,
which may be present in a subject in vivo including a solid tissue
that may be accessed further to a surgical procedure, or that may
be excised, for instance incident to a surgical procedure according
to standard medical practices.
[0122] Solid tissues are well known to the medical arts and may
include any cohesive, spatially discrete non-fluid defined anatomic
compartment that is substantially the product of multicellular,
intercellular, tissue and/or organ architecture, such as a
three-dimensionally defined compartment that may comprise or derive
its structural integrity from associated connective tissue and may
be separated from other body areas by a thin membrane (e.g.,
meningeal membrane, pericardial membrane, pleural membrane, mucosal
membrane, basement membrane, omentum, organ-encapsulating membrane,
or the like). Non-limiting exemplary solid tissues may include
brain, liver, lung, kidney, prostate, ovary, spleen, lymph node
(including tonsil), thyroid, pancreas, heart, skeletal muscle,
intestine, larynx, esophagus and stomach. Anatomical locations,
morphological properties, histological characterization, and
invasive and/or non-invasive access to these and other solid
tissues are all well known to those familiar with the relevant
arts.
[0123] Certain particularly preferred embodiments as disclosed
herein relate to a method for selective delivery of a fluid-phase
agent to a solid tissue. As also noted above, such selective
delivery obviates the need for excessive systemic concentrations of
therapeutic or candidate agents in order to achieve therapeutically
effective concentrations in the desired solid tissue, thereby
avoiding clinically detrimental toxicities to uninvolved tissues
and also avoiding undesirable side-effects. Related embodiments
contemplate the testing of currently non-approved candidate agents
through such selective delivery to a solid tissue. Without wishing
to be bound by theory, according to these embodiments, direct
effects of the candidate agent on the solid tissue (e.g., solid
tumor) can be evaluated by in vivo administration followed by ex
vivo analysis of excised tissue, without threatening the health of
the subject, because the dose used for direct administration into
the solid tissue is far lower than the minimal dose that would
otherwise be administered systemically. (The minimal dose is the
smallest amount of the agent that will produce a desired
physiologic effect in the subject.) Given the minute volumes and
low pressures of the present modes of fluid administration, and
full or partial patency of the solid tissue as a physical property
that promotes retention of the administered fluid (also
determinable by existing methodologies, e.g., by imaging and/or by
use of a detectable label as a tracer), the agent that is
selectively administered to the solid tissue according to the
present disclosure is either undetectable outside the solid tissue,
or if detectable outside the solid tissue, the agent is present at
less (in a statistically significant manner) than the minimal
dose.
[0124] Such considerations pertain in related embodiments, wherein
detection in a solid tissue of an altered physiologic state
subsequent to introducing an agent or a plurality of agents
includes detecting a degree of permeation of the agent(s) through
the solid tissue, detecting a degree of absorption of the agent(s)
in the tissue, detecting a physicochemical effect of the agent(s)
on the tissue, and/or detecting a pharmacological effect of the
agent(s) on the tissue. Assays, including fluorescence assays, of
drug permeation or penetration in solid tissues are known in the
art and have been described (e.g., Kerr et al., 1987 Canc.
Chemother. Pharmacol. 19:1 and references cited therein; Nederman
et al., 1981 In Vitro 17:290; Durand, 1981 Canc. Res. 41:3495;
Durand, 1989 JNCI 81:146; Tunggal et al., 1999 Clin. Canc. Res.
5:1583) and may be configured further according to the present
disclosure, for instance, through the detection in histological
serial sections of a detectable label that has been co-administered
to the solid tissue, prior to excision and sectioning, with an
agent of interest.
[0125] In such embodiments, permeation or penetration refers to the
area of retention of an agent in the solid tissue in the immediate
vicinity of the needle from which the agent was introduced
exclusive of perfusion (entry into and dispersion via any blood
vessel), and may include retention of the agent in extracellular
space or extracellular matrix or in association with a cell
membrane or intracellularly. Permeation may be distinct from a
physicochemical effect, which refers to microscopically detectable
mechanical disruption of tissue that results from the needle
insertion or fluid injection itself, or from non-biological
mechanical or chemical tissue disruption caused by the agent (e.g.,
damage to cell membranes or disintegration of cell-cell junctions).
Pharmacological effects include statistically significant
alterations of a cell or tissue physiological state that are
detectable as consequences of the molecular mechanism of action of
the agent, for example, cytoskeletal reorganization, extension or
withdrawal of cellular processes, or evidence of biological signal
transduction as may be detected using any of a number of known
cytological, biochemical, molecular biological or other read-outs.
Comparison of serial sections may permit distinguishing the nature
of the effect that is detected histologically.
[0126] Particularly preferred embodiments include those in which
the solid tissue comprises a tumor, wherein agent delivery may be
made to, and/or sample retrieval may be made from, the solid tumor.
It will be appreciated by persons familiar with the art from the
disclosure herein that in the course of practicing certain
embodiments described herein, a selected region of a tumor may
comprise the site into which the needles of the presently described
devices are inserted, introduced or otherwise contacted with the
tumor. The region may be selected on any number of bases, including
based on imaging that may be conducted before, during or after a
step of needle insertion, introduction or contacting, or based on
imaging conducted before, during or after excising the solid tissue
from a subject, or based on other criteria including but not
limited to anatomic location, accessibility in the course of a
surgical procedure, degree of vascularization or other
criteria.
[0127] Solid tumors of any type are contemplated as being suitable
for intervention using the devices described herein. In certain
preferred embodiments, the solid tumor may be a benign tumor or a
malignant tumor, which may further be a primary tumor, an invasive
tumor or a metastatic tumor. Certain embodiments contemplate a
solid tumor that comprise one of a prostate cancer cell, a breast
cancer cell, a colon cancer cell, a lung cancer cell, a brain
cancer cell and an ovarian cancer cell, but the invention is not
intended to be so limited and other solid tumor types and cancer
cell types may be used. For example, the tumor may comprise a
cancer selected from adenoma, adenocarcinoma, squamous cell
carcinoma, basal cell carcinoma, small cell carcinoma, large cell
undifferentiated carcinoma, chondrosarcoma and fibrosarcoma, or the
like. As also noted elsewhere herein, art-accepted clinical
diagnostic criteria have been established for these and other
cancer types, such as those promulgated by the U.S. National Cancer
Institute (Bethesda, Md., USA) or as described in DeVita, Hellman,
and Rosenberg's Cancer: Principles and Practice of Oncology (2008,
Lippincott, Williams and Wilkins, Philadelphia/Ovid, N.Y.); Pizzo
and Poplack, Principles and Practice of Pediatric Oncology (Fourth
edition, 2001, Lippincott, Williams and Wilkins, Philadelphia/
Ovid, N.Y.); and Vogelstein and Kinzler, The Genetic Basis of Human
Cancer (Second edition, 2002, McGraw Hill Professional, New York).
Other non-limiting examples of typing and characterization of
particular cancers are described, e.g., in Ignatiadis et al. (2008
Pathobiol. 75:104); Kunz (2008 Curr. Drug Discov. Technol. 5:9);
and Auman et al. (2008 Drug Metab. Rev. 40:303).
[0128] An "altered physiologic state" may be any detectable
parameter that directly relates to a condition, process, pathway,
dynamic structure, state or other activity in a solid tissue (and
in preferred embodiments in a solid tumor) including in a region
thereof or a portion therefrom, further including a biological
sample obtained therefrom, and that permits detection of an altered
(e.g., measurably changed in a statistically significant manner
relative to an appropriate control) structure or function in a
biological sample from a subject or biological source. The methods
of the present invention thus pertain in part to such correlation
where an indicator of altered physiologic state may be, for
example, a cellular or biochemical activity, including as further
non-limiting examples, cell viability, cell proliferation,
apoptosis, cellular resistance to anti-growth signals, cell
motility, cellular expression or elaboration of connective
tissue-degrading enzymes, cellular recruitment of angiogenesis, or
other criteria as provided herein.
[0129] "Altered physiologic state" may refer to any condition or
function where any structure or activity that is directly or
indirectly related to a solid tissue function has been changed in a
statistically significant manner relative to a control or standard,
and may have its origin in direct or indirect interactions between
a solid tissue constituent and an introduced agent, or in
structural or functional changes that occur as the result of
interactions between intermediates that may be formed as the result
of such interactions, including metabolites, catabolites,
substrates, precursors, cofactors and the like.
[0130] Additionally, altered physiologic state may include altered
signal transduction, respiratory, metabolic, genetic, biosynthetic
or other biochemical or biophysical activity in some or all cells
or tissues of a subject or biological source, in preferred
embodiments in some or all cells of a solid tissue, and in more
preferred embodiments in some or all cells of a tumor such as a
solid tumor in a solid tissue. As non-limiting examples, altered
biological signal transduction, cell viability, cell proliferation,
apoptosis, cellular resistance to anti-growth signals, cell
motility, cellular expression or elaboration of connective
tissue-degrading enzymes, cellular recruitment of angiogenesis, or
other criteria including induction of apoptotic pathways and
formation of atypical chemical and biochemical crosslinked species
within a cell, whether by enzymatic or non-enzymatic mechanisms,
may all be regarded as indicative of altered physiologic state.
Certain of these and other non-limiting examples are described in
greater detail herein.
[0131] According to certain presently contemplated embodiments, the
efficacy of a candidate agent may be identified by detecting an
altered physiologic state as provided herein, including by
assessing any of a number of biological parameters characteristic
of a cancer cell such as those reviewed by Hanahan and Weinberg
(2000 Cell 100:57) and in the references cited therein. Therein are
disclosed methodologies for determining the effect of a candidate
agent on one or more traits exhibited by cancer cells, and
detectable by any of a variety of techniques known to the art for
determining one or more of (i) an ability to evade apoptosis, (ii)
acquisition of self-sufficiency in growth signals, (iii)
insensitivity to growth-inhibitory signals, (iv) acquisition of
tissue invasive and metastatic phenotype, (v) unlimited replicative
potential, and (vi) sustained angiogenesis. Persons skilled in the
art are familiar with multiple approaches for detecting the
presence of these alterations of physiologic state, which can be
adapted to a particular excised tumor system. See, e.g., Bonificano
et al. (Eds.) Current Protocols in Cell Biology, 2007 John Wiley
& Sons, NY; Ausubel et al. (Eds.) Current Protocols in
Molecular Biology, 2007 John Wiley & Sons, NY; Coligan et al.
(Eds.), Current Protocols in Immunology, 2007 John Wiley &
Sons, NY; Robinson et al. (Eds), Current Protocols in Cytometry,
2007 John Wiley & Sons, NY. Non-limiting examples of parameters
that may be assayed to identify an altered physiologic state
include assays of cell viability, cell division, apoptosis,
necrosis, cell surface marker expression, cellular activation
state, cellular elaboration of extracellular matrix (ECM)
components or of ECM-degrading enzymes, morphometric analysis,
extension or retraction of cellular processes, cytoskeletal
reorganization, altered gene expression, e.g., by in situ
hybridization of immunohistochemistry (e.g., Shibata et al., 2002
J. Anat. 200:309) intracellular phosphoprotein localization (e.g.,
Gavet et al., 1998 J Cell Sci 111:3333), and the like.
[0132] Selection of agents that are known or candidate oncology
agents is understood and determinable by one skilled in the
relevant arts (see, e.g., Berkowet al., eds., The Merck Manual,
16.sup.th edition, Merck and Co., Rahway, N.J., 1992; Goodman et
al., eds., Goodman and Gilman's The Pharmacological Basis of
Therapeutics, 10.sup.th edition, Pergamon Press, Inc., Elmsford,
N.Y., (2001); DeVita, Hellman, and Rosenberg's Cancer: Principles
and Practice of Oncology (2008, Lippincott, Williams and Wilkins,
Philadelphia/Ovid, N.Y.); Pizzo and Poplack, Principles and
Practice of Pediatric Oncology (Fourth edition, 2001, Lippincott,
Williams and Wilkins, Philadelphia/Ovid, N.Y.); Avery's Drug
Treatment: Principles and Practice of Clinical Pharmacology and
Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins,
Baltimore, Md. (1987), Ebadi, Pharmacology, Little, Brown and Co.,
Boston, (1985); Osolci al., eds., Remington's Pharmaceutical
Sciences, 18.sup.th edition, Mack Publishing Co., Easton, Pa.
(1990); Katzung, Basic and Clinical Pharmacology, Appleton and
Lange, Norwalk, Conn. (1992)). Candidate agents may be selected
from resources that disclose listings of investigational
therapeutics, for instance, the National Institutes of Health
(Bethesda, Md.) which maintains a database of ongoing and planned
clinical trials at its "ClinicalTrials.gov" website.
[0133] Candidate agents for use in screening methods and in methods
of rating candidate agents for development into therapeutic agents
such as a therapeutic agent for treating a solid tumor may be
provided as "libraries" or collections of compounds, compositions
or molecules. Such molecules typically include compounds known in
the art as "small molecules" and having molecular weights less than
10.sup.5 daltons, preferably less than 10.sup.4 daltons and still
more preferably less than 10.sup.3 daltons.
[0134] For example, a plurality of members of a library of test
compounds can be introduced as candidate agents to a region of a
solid tumor of known tumor type in each one or a plurality of
subjects having a tumor of the known tumor type, by distributing
each of the candidate agents to a plurality of positions along an
axis within the region in each subject, and after a selected period
of time (e.g., a range of time, a minimum time period or a specific
time period) the region of solid tumor in which the candidate
agents have been introduced can be imaged or removed from each
subject, and each region compared by detecting an effect (if any)
of each agent on the respective position within the region, for
instance, by determining whether an altered physiologic state is
present as provided herein, relative to positions in the region
that are treated with control agents as provided herein, which
would either produce no effect (negative control) or a readily
detectable effect (positive control).
[0135] Candidate agents further may be provided as members of a
combinatorial library, which preferably includes synthetic agents
prepared according to a plurality of predetermined chemical
reactions performed in a plurality of reaction vessels. For
example, various starting compounds may be prepared employing one
or more of solid-phase synthesis, recorded random mix methodologies
and recorded reaction split techniques that permit a given
constituent to traceably undergo a plurality of permutations and/or
combinations of reaction conditions. The resulting products
comprise a library that can be screened followed by iterative
selection and synthesis procedures, such as a synthetic
combinatorial library of peptides (see e.g., PCT/US91/08694,
PCT/US91/04666, which are hereby incorporated by reference in their
entireties) or other compositions that may include small molecules
as provided herein (see e.g., PCT/US94/08542, EP 0774464, U.S. Pat.
No. 5,798,035, U.S. Pat. No. 5,789,172, U.S. Pat. No. 5,751,629,
which are hereby incorporated by reference in their entireties).
Those having ordinary skill in the art will appreciate that a
diverse assortment of such libraries may be prepared according to
established procedures, and tested for their influence on an
indicator of altered mitochondrial function, according to the
present disclosure.
[0136] Other candidate agents may be proteins (including
therapeutic proteins), peptides, peptidomimetics, polypeptides, and
gene therapy agents (e.g., plasmids, viral vectors, artificial
chromosomes and the like containing therapeutic genes or
polynucleotides encoding therapeutic products, including coding
sequences for small interfering RNA (siRNA), ribozymes and
antisense RNA) which in certain further embodiments may comprise an
operably linked promoter such as a constitutive promoter or a
regulatable promoter, such as an inducible promoter (e.g.,
IPTG-inducible), a tightly regulated promoter (e.g., a promoter
that permits little or no detectable transcription in the absence
of its cognate inducer or derepressor) or a tissue-specific
promoter. Methodologies for preparing, testing and using these and
related agents are known in the art. See, e.g., Ausubel (Ed.),
Current Protocols in Molecular Biology (2007 John Wiley & Sons,
NY); Rosenzweig and Nabel (Eds), Current Protocols in Human
Genetics (esp. Ch. 13 therein, "Delivery Systems for Gene Therapy",
2008 John Wiley & Sons, NY); Abell, Advances in Amino Acid
Mimetics and Peptidomimetics, 1997 Elsevier, N.Y.
[0137] Other candidate agents may be antibodies, including
naturally occurring, immunologically elicited, chimeric, humanized,
recombinant, and other engineered antigen-specific immunoglobulins
and artificially generated antigen-binding fragments and
derivatives thereof, such as single-chain antibodies, minibodies,
Fab fragments, bi-specific antibodies and the like. See, e.g.,
Coligan et al. (Eds.), Current Protocols in Immunology (2007 John
Wiley & Sons, NY); Harlow and Lane, Antibodies: A Laboratory
Manual (1988 Cold Spring Harbor Press, Cold Spring Harbor, N.Y.);
Harlow and Lane, Using Antibodies (1999 Cold Spring Harbor Press,
Cold Spring Harbor, N.Y.).
[0138] Pharmaceutically acceptable carriers for therapeutic use are
well known in the pharmaceutical art, and are described, for
example, in Remingtons Pharmaceutical Sciences, Mack Publishing Co.
(A. R. Gennaro edit. 1985). For example, sterile saline and
phosphate-buffered saline at physiological pH may be used.
Preservatives, stabilizers, dyes and other ancillary agents may be
provided in the pharmaceutical composition. For example, sodium
benzoate, sorbic acid and esters of p-hydroxybenzoic acid may be
added as preservatives. Id. at 1449. In addition, antioxidants and
suspending agents may be used. Id. "Pharmaceutically acceptable
salt" refers to salts of drug compounds derived from the
combination of such compounds and an organic or inorganic acid
(acid addition salts) or an organic or inorganic base (base
addition salts). The agents, including drugs, contemplated for use
herein may be used in either the free base or salt forms, with both
forms being considered as being within the scope of the certain
present invention embodiments.
[0139] The pharmaceutical compositions that contain one or more
agents may be in any form which allows for the composition to be
administered to a patient. According to certain preferred
embodiments the composition will be in liquid form and the route of
administration will comprise administration to a solid tissue as
described herein. The term parenteral as used herein includes
transcutaneous or subcutaneous injections, and intramuscular,
intramedullar and intrasternal techniques.
[0140] The pharmaceutical composition is formulated so as to allow
the active ingredients contained therein to be bioavailable upon
administration of the composition to a subject such as a human
patient. Compositions that will be administered to a patient may
take the form of one or more doses or dosage units, where for
example, a pre-measured fluid volume may comprise a single dosage
unit, and a container of one or more compositions (e.g., drugs) in
liquid form may hold a plurality of dosage units. A dose of a drug
includes all or a portion of a therapeutically effective amount of
a particular drug that is to be administered in a manner and over a
time sufficient to attain or maintain a desired concentration range
of the drug, for instance, a desired concentration range of the
drug in the immediate vicinity of a delivery needle in a solid
tissue, and where the absolute amount of the drug that comprises a
dose will vary according to the drug, the subject, the solid tissue
and other criteria with which the skilled practitioner will be
familiar in view of the state of the medical and pharmaceutical and
related arts. In certain embodiments at least two doses of the drug
may be administered, and in certain other embodiments 3, 4, 5, 6,
7, 8, 9, 10 or more doses may be administered.
[0141] A liquid pharmaceutical composition as used herein, whether
in the form of a solution, suspension or other like form, may
include one or more of the following adjuvants: sterile diluents
such as water for injection, saline solution, preferably
physiological saline, Ringer's solution, saline solution (e.g.,
normal saline, or isotonic, hypotonic or hypertonic sodium
chloride), fixed oils such as synthetic mono or digylcerides which
may serve as the solvent or suspending medium, polyethylene
glycols, glycerin, propylene glycol or other solvents;
antibacterial agents such as benzyl alcohol or methyl paraben;
antioxidants such as ascorbic acid or sodium bisulfite; chelating
agents such as ethylenediaminetetraacetic acid; buffers such as
acetates, citrates or phosphates and agents for the adjustment of
tonicity such as sodium chloride or dextrose. The parenteral
preparation can be enclosed in ampoules, disposable syringes or
multiple dose vials made of glass or plastic. Physiological saline
is a preferred adjuvant. An injectable pharmaceutical composition
is preferably sterile. It may also be desirable to include other
components in the preparation, such as delivery vehicles including
but not limited to aluminum salts, water-in-oil emulsions,
biodegradable oil vehicles, oil-in-water emulsions, biodegradable
microcapsules, hydrogels, and liposomes.
[0142] While any suitable carrier known to those of ordinary skill
in the art may be employed in the pharmaceutical compositions of
this invention, the type of carrier will vary depending on the mode
of administration and whether a conventional sustained drug release
is also desired. For parenteral administration, such as
supplemental injection of drug, the carrier preferably comprises
water, saline, alcohol, a fat, a wax or a buffer. Biodegradable
microspheres (e.g., polylactic galactide) may also be employed as
carriers for the pharmaceutical compositions of this invention.
Suitable biodegradable microspheres are disclosed, for example, in
U.S. Pat. Nos. 4,897,268 and 5,075,109. In this regard, it is
preferable according to certain embodiments that the microsphere be
larger than approximately 25 microns, while other embodiments are
not so limited and contemplate other dimensions. Pharmaceutical
compositions may also contain diluents such as buffers,
antioxidants such as ascorbic acid, low molecular weight (less than
about 10 residues) polypeptides, proteins, amino acids,
carbohydrates including glucose, sucrose or dextrins, chelating
agents such as EDTA, glutathione and other stabilizers and
excipients. Neutral buffered saline or saline mixed with
nonspecific serum albumin are exemplary appropriate diluents.
Preferably, an agent (e.g., a therapeutic drug or a candidate drug)
is formulated as a lyophilizate using appropriate excipient
solutions (e.g., sucrose) as diluents.
[0143] For convenience, elements of various embodiments have been
described as components of a number of discrete assemblies.
However, in practice, elements of some of the assemblies may be
omitted or grouped with other assemblies. Accordingly, the
particular arrangements of embodiments disclosed above do not
impose similar organization on other embodiments or the claims.
[0144] When used to refer to agents delivered from needles, the
term fluid is to be read broadly to read on any substance capable
of flowing through such a needle, including liquids, gases,
colloids, suspended solids, etc.
[0145] The abstract of the present disclosure is provided as a
brief outline of some of the principles of the invention according
to one embodiment, and is not intended as a complete or definitive
description of any embodiment thereof, nor should it be relied upon
to define terms used in the specification or claims. The abstract
does not limit the scope of the claims.
[0146] Elements of the various embodiments described above can be
combined, and further modifications can be made, to provide further
embodiments without deviating from the spirit and scope of the
invention. All of the U.S. patents, U.S. patent application
publications, U.S. patent applications, foreign patents, foreign
patent applications and non-patent publications referred to in this
specification and/or listed in the Application Data Sheet, are
incorporated herein by reference, in their entirety. Aspects of the
embodiments can be modified, if necessary to employ concepts of the
various patents, applications and publications to provide yet
further embodiments.
[0147] These and other changes can be made to the embodiments in
light of the above-detailed description. In general, in the
following claims, the terms used should not be construed to limit
the claims to the specific embodiments disclosed in the
specification, but should be construed to include all possible
embodiments along with the full scope of equivalents to which such
claims are entitled. Accordingly, the claims are not limited by the
disclosure.
* * * * *