U.S. patent application number 10/821330 was filed with the patent office on 2004-12-30 for methods and compositions for enhancing immune response.
Invention is credited to Kedl, Ross M., Miller, Richard L., Ortiz, Ronnie, Stoesz, James D., Tomai, Mark A., Zarraga, Isidro Angelo Eleazar.
Application Number | 20040265351 10/821330 |
Document ID | / |
Family ID | 33545722 |
Filed Date | 2004-12-30 |
United States Patent
Application |
20040265351 |
Kind Code |
A1 |
Miller, Richard L. ; et
al. |
December 30, 2004 |
Methods and compositions for enhancing immune response
Abstract
Methods and compositions for enhancing the immune response to an
IRM compound by depositing within a localized tissue region an IRM
depot preparation that provides an extended residence time of
active IRM within the localized tissue region.
Inventors: |
Miller, Richard L.;
(Maplewood, MN) ; Tomai, Mark A.; (Woodbury,
MN) ; Kedl, Ross M.; (Denver, CO) ; Zarraga,
Isidro Angelo Eleazar; (Minneapolis, MN) ; Ortiz,
Ronnie; (Apple Valley, MN) ; Stoesz, James D.;
(Inver Grove Heights, MN) |
Correspondence
Address: |
3M INNOVATIVE PROPERTIES COMPANY
PO BOX 33427
ST. PAUL
MN
55133-3427
US
|
Family ID: |
33545722 |
Appl. No.: |
10/821330 |
Filed: |
April 9, 2004 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
10821330 |
Apr 9, 2004 |
|
|
|
10640904 |
Aug 14, 2003 |
|
|
|
60533703 |
Dec 31, 2003 |
|
|
|
60462140 |
Apr 10, 2003 |
|
|
|
60515256 |
Oct 29, 2003 |
|
|
|
60515604 |
Oct 30, 2003 |
|
|
|
60545424 |
Feb 18, 2004 |
|
|
|
60545542 |
Feb 18, 2004 |
|
|
|
Current U.S.
Class: |
424/423 |
Current CPC
Class: |
Y02A 50/30 20180101;
A61K 31/44 20130101; A61K 47/543 20170801; Y02A 50/489 20180101;
A61K 47/6911 20170801; A61K 39/39 20130101; Y02A 50/41 20180101;
Y02A 50/488 20180101; A61K 39/00 20130101 |
Class at
Publication: |
424/423 |
International
Class: |
A61K 031/44 |
Claims
What is claimed is:
1. A method of enhancing the immune response to an IRM compound,
comprising: depositing within a localized tissue region an IRM
depot preparation that provides an extended residence time within
the localized tissue region.
2. The method of claim 1, wherein the localized tissue region is a
breast cancer tumor.
3. The method of claim 1, wherein the localized tissue region is a
stomach cancer tumor.
4. The method of claim 1, wherein the localized tissue region is a
lung cancer tumor.
5. The method of claim 1, wherein the localized tissue region is a
head or neck cancer tumor.
6. The method of claim 1, wherein the localized tissue region is a
colorectal cancer tumor.
7. The method of claim 1, wherein the localized tissue region is a
renal cell carcinoma tumor.
8. The method of claim 1, wherein the localized tissue region is a
pancreatic cancer tumor
9. The method of claim 1, wherein the localized tissue region is a
basal cell carcinoma tumor
10. The method of claim 1, wherein the localized tissue region is a
cervical cancer tumor
11. The method of claim 1, wherein the localized tissue region is
melanoma cancer tumor.
12. The method of claim 1, wherein the localized tissue region is
prostate cancer tumor.
13. The method of claim 1, wherein the localized tissue region is
ovarian cancer tumor.
14. The method of claim 1, wherein the localized tissue region is
bladder cancer tumor.
15. The method of claim 1, wherein the localized tissue region is
viral-infected lesion or organ.
16. The method of claim 1, wherein the localized tissue region is
includes a vaccine.
17. The method of claim 1, wherein the localized tissue region is a
particular organ subject to a disease that is treatable using the
IRM compound.
18. The method of claim 1, wherein the IRM depot preparation
comprises a lipid-modified IRM.
19. The method of claim 1, wherein the IRM depot preparation
comprises an IRM compound attached to support material.
20. The method of claim 1, wherein the IRM depot preparation
comprises solid particles of IRM compound.
21. The method of claim 1, wherein the IRM depot preparation
comprises an emulsion.
22. The method of claim 1, wherein the IRM depot preparation
comprises micelles.
23. The method of claim 1, wherein the IRM depot preparation
comprises IRM within a biodegradable polymer matrix.
24. The method of claim 1, wherein the IRM depot preparation
comprises IRM compound incorporated into lipid membranes, lipid
vesicles, or liposomes.
25. The method of claim 1, wherein the IRM depot preparation
provides pulsed delivery of an IRM compound.
26. The method of claim 1, wherein the IRM depot preparation
comprises an osmotically driven cylinder.
27. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using needle
injection.
28. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using surgical
implantation.
29. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using laparoscopic
implantation.
30. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using catheter
implantation.
31. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using a microneedle
array.
32. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using high-velocity
particle implantation.
33. The method of claim 1, wherein the IRM depot preparation is
delivered within the localized tissue region using an image guiding
technique selected from ultrasound, MRI, or real-time X-ray
fluoroscopy.
34. The method of claim 1 wherein the IRM is an agonist of at least
one TLR selected from the group consisting of TLR6, TLR7, TLR8,
TLR9 and combinations thereof.
35. The method of claim 1 wherein the IRM is a selective TLR
agonist of TLR7.
36. The method of claim 1 wherein the IRM is a selective TLR
agonist of TLR8.
37. The method of claim 1 wherein the IRM is a selective TLR
agonist of TLR9.
38. The method of claim 1 wherein the IRM is a TLR agonist of both
TLR 7 and 8.
39. The method of claim 1 wherein the IRM is a small molecule
immune response modifier.
40. The method of claim 1 wherein the IRM is selected from the
group consisting of imidazoquinoline amines including, but not
limited to, amide substituted imidazoquinoline amines, sulfonamide
substituted imidazoquinoline amines, urea substituted
imidazoquinoline amines, aryl ether substituted imidazoquinoline
amines, heterocyclic ether substituted imidazoquinoline amines,
amido ether substituted imidazoquinoline amines, sulfonamido ether
substituted imidazoquinoline amines, urea substituted
imidazoquinoline ethers, and thioether substituted imidazoquinoline
amines; tetrahydroimidazoquinoline amines including, but not
limited to, amide substituted tetrahydroimidazoquinoline amines,
sulfonamide substituted tetrahydroimidazoquinoline amines, urea
substituted tetrahydroimidazoquinoline amines, aryl ether
substituted tetrahydroimidazoquinoline amines, heterocyclic ether
substituted tetrahydroimidazoquinoline amines, amido ether
substituted tetrahydroimidazoquinoline amines, sulfonamido ether
substituted tetrahydroimidazoquinoline amines, urea substituted
tetrahydroimidazoquinoline ethers, and thioether substituted
tetrahydroimidazoquinoline amines; imidazopyridine amines
including, but not limited to, amide substituted imidazopyridines,
sulfonamido substituted imidazopyridines, and urea substituted
imidazopyridines; 1,2-bridged imidazoquinoline amines; 6,7-fused
cycloalkylimidazopyridine amines; imidazonaphthyridine amines;
tetrahydroimidazonaphthyridine amines; oxazoloquinoline amines;
thiazoloquinoline amines; oxazolopyridine amines; thiazolopyridine
amines; oxazolonaphthyridine amines; thiazolonaphthyridine amines;
pharmaceutically acceptable salts thereof; and combinations
thereof.
41. The method of claim 1 wherein the IRM is selected from the
group consisting of amide substituted imidazoquinoline amines,
sulfonamide substituted imidazoquinoline amines, urea substituted
imidazoquinoline amines, aryl ether substituted imidazoquinoline
amines, heterocyclic ether substituted imidazoquinoline amines,
amido ether substituted imidazoquinoline amines, sulfonamido ether
substituted imidazoquinoline amines, urea substituted
imidazoquinoline ethers, and thioether substituted imidazoquinoline
amines; tetrahydroimidazoquinoline amines including, but not
limited to, amide substituted tetrahydroimidazoquinoli- ne amines,
sulfonamide substituted tetrahydroimidazoquinoline amines, urea
substituted tetrahydroimidazoquinoline amines, aryl ether
substituted tetrahydroimidazoquinoline amines, heterocyclic ether
substituted tetrahydroimidazoquinoline amines, amido ether
substituted tetrahydroimidazoquinoline amines, sulfonamido ether
substituted tetrahydroimidazoquinoline amines, urea substituted
tetrahydroimidazoquinoline ethers, and thioether substituted
tetrahydroimidazoquinoline amines; imidazopyridine amines
including, but not limited to, amide substituted imidazopyridines,
sulfonamido substituted imidazopyridines, and urea substituted
imidazopyridines; 1,2-bridged imidazoquinoline amines; 6,7-fused
cycloalkylimidazopyridine amines; imidazonaphthyridine amines;
tetrahydroimidazonaphthyridine amines; oxazoloquinoline amines;
thiazoloquinoline amines; oxazolopyridine amines; thiazolopyridine
amines; oxazolonaphthyridine amines; thiazolonaphthyridine amines;
pharmaceutically acceptable salts thereof; and combinations
thereof.
42. The method of claim 1, wherein the IRM comprises a
2-aminopyridine fused to a five membered nitrogen-containing
heterocyclic ring.
43. The method of claim 1, wherein the IRM depot preparation
comprises a CpG IRM.
44. The method of claim 1 wherein the IRM depot preparation further
comprises one or more additional active ingredients.
45. The method of claim 28, wherein the active ingredient comprises
a vaccine.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a Continuation-In-Part of U.S.
patent application Ser. No. 10/640,904, filed on Aug. 14, 2003, and
claims priority to U.S. Provisional Patent Application Ser. Nos.
60/533,703, filed Dec. 31, 2003, 60/462,140, filed on Apr. 10,
2003, 60/545,424, filed on Feb. 18, 2004, 60/515,256, filed on Oct.
29, 2003, 60/545,542, filed on Feb. 18, 2004, and U.S. 60/515,604,
filed Oct. 30, 2003 each of which is incorporated herein by
reference in their entirety.
BACKGROUND
[0002] There has been a major effort in recent years, with
significant successes, to discover new drug compounds that act by
stimulating certain key aspects of the immune system, as well as by
suppressing certain other aspects (see, e.g., U.S. Pat. Nos.
6,039,969 and 6,200,592). These compounds, sometimes referred to as
immune response modifiers (IRMs), appear to act through basic
immune system mechanisms known as toll-like receptors to induce
selected cytokine biosynthesis and may be used to treat a wide
variety of diseases and conditions. For example, certain IRMs may
be useful for treating viral diseases (e.g., human papilloma virus,
hepatitis, herpes), neoplasias (e.g., basal cell carcinoma,
squamous cell carcinoma, actinic keratosis), and TH2-mediated
diseases (e.g., asthma, allergic rhinitis, atopic dermatitis), and
are also useful as vaccine adjuvants. Unlike many conventional
anti-viral or anti-tumor compounds, the primary mechanism of action
for IRMs is indirect, by stimulating the immune system to recognize
and take appropriate action against a pathogen.
[0003] Many of the IRM compounds are small organic molecule
imidazoquinoline amine derivatives (see, e.g., U.S. Pat. No.
4,689,338), but a number of other compound classes are now known as
well (see, e.g., U.S. Pat. No. 5,446,153) and more are still being
discovered. Other IRMs have higher molecular weights, such as
oligonucleotides, including CpGs (see, e.g., U.S. Pat. No.
6,194,388). In view of the great therapeutic potential for IRMs,
and despite the important work that has already been done, there is
a substantial ongoing need for new means of controlling the
delivery and activity of IRMs in order to expand their uses and
therapeutic benefits.
SUMMARY
[0004] It has been found that many immune response modifier (IRM)
compounds (described infra) often have a relatively short half-life
in terms of residence time at a given delivery location, typically
less than about 1-2 hours for small molecule IRMs. They appear to
be cleared, metabolized, or simply diffuse away from within a local
delivery site rather easily in many cases. This short residence
duration may reduce the IRM's ability to activate some immune
system cells at the desired site. Hence, it is now believed that
the effectiveness of IRM compounds may be enhanced by maintaining a
depot of active IRM compound within a localized region of tissue
for an extended period. Importantly, not only do IRMs have the
ability to modulate the immune system locally, but by inducing
certain chemokines, such as, e.g., MIP-3.alpha., MIP-1.alpha.,
IP-10, they can recruit additional critical immune system cells,
such as dendritic cells, to the localized tissue region. For
example, Furumoto et al., J. Clin Invest, March 2004, no.
113(5):774-83, discusses the use of CCL20/MIP-3alpha and CpGs to
recruit dendritic cells, although apparently not in a depot. But by
maintaining the IRM present for an extended period within the
localized tissue region, the IRM can further activate the immune
system cells that have been recruited to the localized site, thus
creating a further synergistic effect.
[0005] The IRM depoting methods and compositions of the present
invention can thus provide important additional time for activation
and/or infiltration of responsive immune system cells (e.g.,
dendritic cells, monocytes/macrophages, and B cells) within a
specific localized tissue region. Moreover, these methods and
formulations may also help assure that the immune response is
correctly targeted to an immunogen at the intended desired tissue
location (e.g., where there are neoplastic cells, virus infected
cells, or vaccine antigen present). This later point--the ability
to target by co-locating the IRM, antigen presenting cells (APCs),
and antigen--is surprisingly important because it may enhance
recognition by the immune system of the targeted disease antigens,
and may also reduce the potential for unwanted immune system
stimulation away from the actual disease target.
[0006] It is also believed that IRM depot preparations that provide
a pulsed IRM delivery (i.e., where the active IRM is release
intermittently in pulses over time) may be particularly desirable
for certain applications.
[0007] Accordingly, the invention includes a method of enhancing
the immune response to an IRM compound, comprising depositing
within a localized tissue region an IRM depot preparation that
provides an extended residence time within the localized tissue
region. This contrasts with either injection of a simple solution
or topical delivery via cream, gel, or patch. The invention also
includes IRM depot preparations disclosed herein, and methods of
treatment using the IRM depot preparations disclosed herein.
[0008] The IRM localized tissue region may be, e.g., a cancer,
infected lesion or organ, or vaccination site. The localized tissue
region may be, e.g., a breast cancer tumor, stomach cancer tumor,
lung cancer tumor, head or neck cancer tumor, colorectal cancer
tumor, renal cell carcinoma tumor, pancreatic cancer tumor, basal
cell carcinoma tumor, pancreatic cancer tumor, cervical cancer
tumor, melanoma cancer tumor, prostate cancer tumor, ovarian cancer
tumor, or bladder cancer tumor. The localized tissue region may
include a vaccine.
[0009] The IRM depot preparation may comprise, e.g., a
lipid-modified IRM, an IRM compound attached to support material,
solid particles of IRM compound, an emulsion, micelles, an IRM
within a biodegradable polymer matrix, IRM compound incorporated
into lipid membranes, lipid vesicles, or liposomes.
[0010] The IRM depot preparation may provide pulsed delivery of an
IRM compound to the localized tissue region. The IRM depot
preparation may comprise an osmotically driven cylinder. The IRM
depot preparation may be delivered within the localized tissue
region using, e.g., needle injection, surgical implantation,
laparoscopic implantation, catheter implantation, a microneedle
array, or high-velocity particle implantation.
[0011] The IRM may be an agonist of at least one TLR selected from
the group consisting of TLR6, TLR7, TLR8, TLR9 and combinations
thereof. The IRM may be a selective TLR agonist of TLR 7, TLR 8, or
TLR 9, or an agonist of both TLR 7 and 8. Many of the IRM compounds
disclosed herein are TLR 7 and/or 8 agonists. The IRM may
alternatively be a TLR 4 agonist. The IRM may be preferably be a
small molecule immune response modifier, for example comprising a
2-aminopyridine fused to a five membered nitrogen-containing
heterocyclic ring.
[0012] The term "comprises" and variations thereof do not have a
limiting meaning where these terms appear in the description and
claims, As used herein, "a," "an," "the," "at least one," and "one
or more" are used interchangeably.
[0013] As used herein, "treating" a condition or a subject includes
therapeutic, prophylactic, and diagnostic treatments.
[0014] Also herein, the recitations of numerical ranges by
endpoints include all numbers subsumed within that range (e.g., 1
to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).
[0015] The above summary of the present invention is not intended
to describe each disclosed embodiment or every implementation of
the present invention. The description that follows more
particularly exemplifies illustrative embodiments. In several
places throughout the application, guidance is provided through
lists of examples, which examples can be used individually and in
various combinations. In each instance, the recited list serves
only as a representative group and should not be interpreted as an
exclusive list.
[0016] Various other features and advantages of the present
invention should become readily apparent with reference to the
following detailed description, examples, claims and appended
drawings. In several places throughout the specification, guidance
is provided through lists of examples. In each instance, the
recited list serves only as a representative group and should not
be interpreted as an exclusive list.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS OF THE
INVENTION
[0017] The present invention is directed to methods and
formulations of immune response modifiers (IRMs) that can be
deposited within a localized tissue region and provide locally
active IRM compounds for an extended period of time. One way this
can be described is in terms of the IRM half-life within a
localized tissue region. To illustrate, if a conventional solution
formulation of a given IRM compound is injected into a solid tumor
so as to achieve a resulting tissue concentration of active IRM
within the tumor, the concentration may be about half only two
hours later. This would be considered an IRM residence half-life of
about 2 hours, although the rate of IRM clearance may not always be
constant. By contrast, if an IRM depot preparation such as those
described herein is injected into a localized tissue region, such
as solid tumor, so as to achieve a desired IRM concentration, the
concentration of active IRM in the tumor tissue (localized tissue
region) may not be down to half until, e.g., 10-14 days later. This
would be considered an IRM residence half-life of about 2
weeks.
[0018] The present invention thus provides active IRMs within a
localized tissue region for a time longer than a comparable
concentration of the IRM in a conventional solution, wherein at
least about 50% of the IRM compound delivered via the IRM depot
preparation remains localized at the treatment site for more than
at least about 2 hours. For example, the IRM residence half-life
may be at least 12 hours, 24 hours, 7 days, two weeks, a month, or
even several months.
[0019] As described below, the benefits of the present invention in
terms of enhanced immune response and/or better targeting of the
immune system to intended antigens can be accomplished with many
different IRM depot preparations, IRM compounds, optionally with
other active agents, and can be delivered to various localized
tissue regions for a wide range of treatments.
[0020] IRM Depot Preparations
[0021] As used herein, IRM depot preparation refers to compositions
that provide active IRM compound for an extended period to a
localized tissue region (as opposed to an extended release IRM
formulation for providing extended systemic delivery, although that
may use a drug depot for systemic delivery).
[0022] There are at least two general ways of maintaining a
localized IRM depot effect. The IRM may either be attached to some
other material that helps hold the IRM in place within the desired
localized tissue region, or the IRM may be released over time from
a controlled release formulation in such a way that active IRM is
present locally at a desired concentration for an extended
period.
[0023] Examples of attaching an IRM to another material that can be
used in an IRM depot preparation include the IRM-support complexes
disclosed in, e.g., copending applications U.S. 60/462,140,
60/515,256, and US 2003/25523.
[0024] Examples of controlled release formulations and methods that
can be used in an IRM depot preparation, although typically used
for extended release systemic drug delivery, are disclosed in,
e.g., U.S. Pat. No. 6,126,919 (biocompatible compounds), O'Hagan
and Singh, Microparticles as vaccine adjuvants and delivery
systems, Expert Rev. Vaccines 2(2), p. 269-83 (2003), and Vogel et
al., A Compendium of Vaccine Adjuvants and Excipients, 2.sup.nd
Edition, Bethesda, Md.: National Institute of Allergy &
Infectious Diseases, 1998 (available at www.niaid.nih.gov/hivva-
ccines/pdf/compendium.pdf). Also, it is usually desired to prevent,
or at least reduce the occurrence of, the systemic distribution of
the IRM after it leaves the localized tissue region. One way to
facilitate this is to select an IRM that is metabolized or cleared
rapidly once the IRM leaves the localized tissue region.
[0025] Some general examples of IRM depot preparations that can
provide an increased IRM residence time within a localized tissue
region include but are not limited to the following:
[0026] 1. The IRM compound may be attached (e.g., conjugated,
coated, or ion-paired) onto to other support materials, such as
plastic, metal, minerals (e.g., alum), or silicone, in the form of
particles, beads, fibers, meshes, polymers, etc., as disclosed in,
e.g., copending applications U.S. 60/462,140, 60/515,256, and US
2003/25523. These IRM support complexes can then be deposited
within a desired tissue site and remain in place and active for an
extended period of time. This a highly versatile approach in part
because the IRM compounds can be attached to many different support
complex materials, and because the IRM can remain active even while
they remain attached to the support complex material.
[0027] For example, IRM, and an antigen if desired, may adsorb onto
the surface of alum particles to enhance antigen presentation and
endocytosis of particulates. Another example would where the IRM is
covalently linked to a polymer backbone through a link that is
subject to hydrolysis or enzymatic activity at a slow, controlled
rate.
[0028] 2. The IRM compound may be conjugated directly to a lipid
group, as disclosed in, e.g., copending application U.S.
60/515,604, which in itself can provide an IRM depot preparation.
These lipid-modified or lipidated IRMs may also be used as the IRM
compound in the other IRM depot preparations described herein,
e.g., for formation of suspensions, incorporation into emulsions,
lipid membranes, lipid vesicles, liposomes, and the like.
[0029] For example, if a lipidated IRM is in suspension,
formulations of which are described below, is injected
subcutaneously at 10 mg/kg (200 ug of drug in a normal B6 mouse), a
substantial amount of the IRM depot preparation is still visible
under the skin 10-14 days later.
[0030] 3. The IRM compound may be used in the form of solid IRM
particles, where the particles may have a limited solubility so
that once implanted within the localized tissue region they
dissolve slowly over an extended period. This contrasts with the
situation where solid drug particle suspensions are delivered that
then dissolve relatively quickly upon delivery (e.g., within an
hour). The IRM particles may be amorphous or crystalline and in the
form of fine powders, liquid suspensions, such as colloidal
suspensions, or may be included within gels or creams, and the
like. The solid IRM particles may be essentially pure IRM compound,
or may include carriers or fillers. The average size of the
particles may be less than 1 micron, or from about 1-100 microns,
or larger than 100 microns, depending on the particular IRM used
and the desired release characteristics. An average particle size
of between 1-20 microns will often be suitable. When introduced
within a localized tissue region the solid IRM particles can slowly
release active IRM compound the local area, providing extended
residence times. The rate of release will depend on solubility of
the particular IRM used, which may be influenced by such things as
selection of polymorph forms, salts, and stereoisomers, in addition
to the physicochemical properties of any carriers or fillers, if
used.
[0031] For example, a colloidal IRM suspension may be formed using
an IRM dissolved in a water-miscible organic injectable solvent
(e.g. N-methyl pyrrolidone or NMP) (compartment 1). Then, an
antigen, if desired, may be dissolved in an aqueous solution with
surfactants (e.g. Tween 80) (compartment 2). Prior to
administration to a localized tissue region, such as subcutaneous
injection, colloidal particles of IRM are formed by mixing
compartment 1 into compartment 2, causing precipitation of IRM into
fine particles. Of course, where appropriate an IRM colloidal
suspension can be made prior to packaging, with instructions for
shaking/vortexing prior to administration.
[0032] 4. The IRM compound may be encapsulated, incorporated, or
dissolved into biodegradable polymeric matrices such as poly lactic
acid (polylactides), poly glycolic acid (polyglycolides),
poly(d,l-lactide-co-glycolide) (PLGA), polyorthoesters,
polyanhydrides, polyphosphazenes, and polyurethanes. Such
biodegradable matrices are often used to provide extended release
systemic drug delivery, but can also be adapted for use to provide
extended IRM delivery within a localized tissue region, for example
directly within a tumor mass, infection site, or vaccination
site.
[0033] For example, an IRM compound and a vaccine antigen may be
dissolved in a polymer solvent like N-methyl pyrrolidone or NMP
(compartment 1). A selected polymer may then be dissolved in the
solvent (compartment 2). Compartment 1 and 2 are mixed upon
administration, e.g., using a double cartridge syringe with a
static mixer. When injected, the solvent (e.g. NMP) which is
miscible with water, diffuses away, leaving a semi-solid implant
containing both IRM and Antigen.
[0034] 5. The IRM compound may be incorporated into single
emulsions such as oil-in-water (o/w) or water-in-oil (w/o), or
multiple emulsions such as water-in-oil-in water (w/o/w) and
oil-in-water-in-oil (o/w/o). Antigen can also be incorporated into
the emulsion in addition to the IRM (e.g. to generate a more
specific immune response). The IRM and antigen can partition
between the oil and water phases, or lie on the discrete phase
(e.g. oil droplet) surface. The emulsion format may act
synergistically with the IRM for improved immune response (e.g.
antigen on oil droplet surface may enhance its presentation to
cells of the immune system, while IRM can enhance uptake of the
antigen on the oil droplet by cells of the immune system.
[0035] For example, an o/w emulsion may use an IRM in a MF59-based
emulsion containing squalene (oil-phase) and surfactant (e.g.,
Tween 80, Span 85), and water. The IRM may be pre-dissolved in the
water phase or the oil phase. Another example is to use a vegetable
oil (sesame oil, soybean oil, mineral oil, e.g. emulsion based on
Freund's Incomplete Adjuvant, etc.) with IRM and surfactant (e.g.,
lecithin) in water. A w/o emulsion may use an IRM dissolved in
water, surfactant (e.g., mannide monooleate) and mineral oil. A w/o
emulsion can also be made using water with IRM dissolved, and
injectable vegetable oils with appropriate emulsifiers and
surfactants (Tween 80, Cremophore EL, etc.). Antigen may also be
pre-dissolved in water or the oil phase and incorporated in the
emulsion.
[0036] 6. The IRM compound may be incorporated into lipid
membranes, lipid vesicles, and liposomes. Within lipsome
preparations, the IRM may be loaded into the membrane, on the
membrane surface, or in the liposome core. For example, IRM
liposomes may use DOTAP transfection agent and cholesterol to
entrap IRM, and antigen if desired, in a liposome.
[0037] 7. The IRM compound may be delivered to the localized tissue
region using an osmotically driven cylinder implanted within the
tissue.
[0038] 8. The IRM compound may be incorporated into a bioadhesive
polymer such as a hydrogel, including, for example, those described
by H. S. Sawhney, C. P. Pathak and J. A. Hubell in Macromolecules,
(1993) 26:581-587, as well as polyhyaluronic acids, casein,
polysaccharides, keratin, collagen, gelatin, glutin, polyethylene
glycol, crosslinked albumin, fibrin, polyanhydrides, polyacrylic
acids, alginate, chitosan, poly(methyl methacrylates), poly(ethyl
methacrylates), poly(butyl methacrylate), poly(isobutyl
methacrylate), poly(hexyl methacrylate), poly(isodecyl
methacrylate), poly(lauryl methacrylate), poly(phenyl
methacrylate), poly(methyl acrylate), poly(isopropyl acrylate),
poly(isobutyl acrylate), poly(octadecyl acrylate), and cellulose
gums. Alternatively, polymeric hydrogel materials can be
constructed from poly(vinyl alcohol) precursors as disclosed in
U.S. Pat. Nos. 4,528,325 and 4,618,649 or from poly(methyl
methacrylate). Poly(methyl methacrylate) is commercially available
and is often used in ophthalmic devices such as intraocular lenses,
contact lenses, and the like.
[0039] A suitable hydrogel can be natural, synthetic, or a
combination thereof. In some embodiments, the hydrogel can be
thermally responsive to a designed temperature such as, for
example, a hydrogel as described in U.S. patent application Ser.
No. 10/626,261, filed Jul. 24, 2003. For example, the thermally
responsive hydrogels can be harden when they are warmed up to body
temperature, can be further harden upon UV irradiation.
[0040] Bioadhesive organic polymers are preferred for certain
applications of IRMs. For example, if the IRM is to be used for
treating bladder cancer, a bioadhesive polymer may be desired.
Advantageously, the adhesive qualities of the formulation would
allow the IRM to be in contact with the biological tissue allowing
for greater contact time for cytokine induction.
[0041] It should also be noted that where solid particles are
involved, the particles may be in any number of forms, e.g.,
irregular particulates, spheres, plates, flakes, rods or other
shapes, and they may be porous or non-porous. Particles can be
lyophilized, then for example provided with a diluent to create a
microsuspension prior to administration. For vaccines, antigen may
be encapsulated within a particle matrix, for example a
biodegradable polymer, and IRM compound either incorporated into or
adsorbed on surface of the particle, either by physical or chemical
adsorption.
[0042] The IRM depot preparations may provide IRM compound to the
desired localized tissue region with various IRM residence
half-life times, generally of at least 2 hours. For example, the
IRM residence half-life may be at least 12 hours, 24 hours, 7 days,
two weeks, a month, or even several months.
[0043] Further, the IRM depot preparation can be designed to
achieve constant or pulsed delivery to the localized tissue region.
Pulsed delivery may be desirable in order to provide intermittent
dosing of an IRM to the local tissue region over time. For example,
a combination of biodegradable polymers can be used that have
differing degradation, and thus IRM release, rates. The depot
preparation may contain a homogeneous mixture of various
biodegradable polymers, or the polymers may be utilized in a
segmented fashion to achieve complex degradation profiles. The
depot preparation may also be coated with various polymers to
achieve zero-order, first-order, and pulsatile IRM release. If in
the form of particles, some particles may use a polymer matrix that
releases the IRM (and other optional ingredients, such as vaccine
antigen) over 24 hours, other particles that release about two
weeks later, and so on. It may be particularly desirable for
vaccine purposes to provide continuous release of an antigen and
use an IRM depot preparation that provides pulsatile release of the
IRM compound. Also, the IRM release timing may either be regular,
e.g., initially and once weekly for several weeks, or it may be
irregular, e.g., initially and then 3 days, 2 weeks, and 2 months
apart.
[0044] The IRM depot preparations may be delivered into a desired
localized tissue region via any suitable route, e.g., including but
not limited to a subcutaneous, intradermal, intramuscular,
intrathecal, intra-organ, intratumoral, intralesional,
intravesicle, and intraperitoneal route of delivery. A "localized
tissue region" will generally be a relatively small portion of the
body, e.g., less than 10% by volume, and often less than 1% by
volume. For example, depending on the size of, e.g., a solid tumor
or cancerous organ, the localized tissue region will typically be
on the order of no more than about 500 cm.sup.3, often less than
about 100 cm.sup.3, and in many instances 10 cm.sup.3 or less. For
some applications the localized tissue region will be 1 cm.sup.3 or
less (e.g., for small tumor nodules, viral lesions, or vaccination
sites). However, in certain instances the localized tissue region
may be a particularly large region, up to several liters, for
example to treat metastasized cancer within the entire peritoneal
cavity (e.g., using an IRM depot preparation to retain the IRM for
an extended time within the peritoneal cavity). The IRM depot
preparations may be delivered using, e.g., needle injection,
surgical, laparoscopic, or catheter implantation, microneedle
array, high-velocity particle implantation, or any other known
method for introducing a preparation into a localized tissue
region. Delivery to the localized tissue region may be in
conjunction with image guiding techniques using, for example,
ultrasound, MRI, real-time X-ray (fluoroscopy), etc.
[0045] Additional Agents
[0046] In addition to one or more IRM compounds, the IRM depot
preparations and methods of the present invention can include
additional agents. Alternatively, the additional agent(s) can be
administered separately from the IRM depot preparation.
[0047] Such additional agents may be additional drugs, including,
for example, a vaccine or a tumor necrosis factor receptor (TNFR)
agonist. Vaccines include any material that raises either humoral
and/or cell mediated immune response, such as live or attenuated
viral and bacterial immunogens and inactivated viral,
tumor-derived, protozoal, organism-derived, fungal, and bacterial
immunogens, toxoids, toxins, polysaccharides, proteins,
glycoproteins, peptides, cellular vaccines, such as using dendritic
cells, DNA vaccines, recombinant proteins, glycoproteins, and
peptides, and the like, for use in connection with, e.g., cancer
vaccines, BCG, cholera, plague, typhoid, hepatitis A, B, and C,
influenza A and B, parainfluenza, polio, rabies, measles, mumps,
rubella, yellow fever, tetanus, diphtheria, hemophilus influenza b,
tuberculosis, meningococcal and pneumococcal vaccines, adenovirus,
HIV, chicken pox, cytomegalovirus, dengue, feline leukemia, fowl
plague, HSV-1 and HSV-2, hog cholera, Japanese encephalitis,
respiratory syncytial virus, rotavirus, papilloma virus, severe
acute respiratory syndrome (SARS), anthrax, and yellow fever. See
also, e.g., vaccines disclosed in WO 02/24225. Such additional
agents can include, but are not limited to, drugs, such as
antiviral agents or cytokines. The vaccine may be separate or may
be physically or chemically linked to the IRM, such as by chemical
conjugation or other means, so that they are delivered as a unit.
TNFR agonists that may be delivered in conjunction with the IRM
depot preparation include, but are not limited to, CD40 receptor
agonists, such as disclosed in copending application U.S.
60/437,398. Other active ingredients for use in combination with an
IRM depot preparation of the present invention include those
disclosed in, e.g., US 2003/0139364.
[0048] Immune Response Modifier Compounds:
[0049] Immune response modifiers ("IRM") useful in the present
invention include compounds that act on the immune system by
inducing and/or suppressing cytokine biosynthesis. IRM compounds
possess potent immunostimulating activity including, but not
limited to, antiviral and antitumor activity, and can also
down-regulate other aspects of the immune response, for example
shifting the immune response away from a TH-2 immune response,
which is useful for treating a wide range of TH-2 mediated
diseases. IRM compounds can also be used to modulate humoral
immunity by stimulating antibody production by B cells. Further,
various IRM compounds have been shown to be useful as vaccine
adjuvants (see, e.g., U.S. Pat. Nos. 6,083,505, U.S. Pat. No.
6,406,705, and WO 02/24225).
[0050] In particular, certain IRM compounds effect their
immunostimulatory activity by inducing the production and secretion
of cytokines such as, e.g., Type I interferons, TNF-.alpha., IL-1,
IL-6, IL-8, IL-10, IL-12, IP-10, MIP-1, MIP-3, and/or MCP-1, and
can also inhibit production and secretion of certain TH-2
cytokines, such as IL-4 and IL-5. Some IRM compounds are said to
suppress IL-1 and TNF (U.S. Pat. No. 6,518,265).
[0051] For some embodiments, the preferred IRM compounds are
so-called small molecule IRMs, which are relatively small organic
compounds (e.g., molecular weight under about 1000 daltons,
preferably under about 500 daltons, as opposed to large biologic
protein, peptides, and the like). Although not bound by any single
theory of activity, some IRMs are known to be agonists of at least
one Toll-like receptor (TLR). IRM compounds that are agonists for
TLRs selected from 6, 7, 8, and/or 9 may be particularly useful for
certain applications. In some applications, for example, the
preferred IRM compound is not a TLR7 agonist and is a TLR8 or TLR9
agonist. In other applications, for example, the IRM is a TLR7
agonist and is not a TLR 8 agonist. Some small molecule IRM
compounds are agonists of TLRs such as 6, 7, and/or 8, while
oligonucleotide IRM compounds are agonists of TLR9, and perhaps
others. Thus, in some embodiments, the IRM that is included in the
IRM delivery apparatus may be a compound identified as an agonist
of one or more TLRs.
[0052] For example, without being bound to any particular theory or
mechanism of action, IRM compounds that activate a strong cytotoxic
lymphocyte (CTL) response may be particularly desirable as vaccine
adjuvants, especially for therapeutic viral and/or cancer vaccines
because a therapeutic effect in these settings is dependent on the
activation of cellular immunity. For example, studies have shown
that activation of T cell immunity in a given patient has a
significant positive effect on the prognosis of the patient.
Therefore the ability to enhance T cell immunity is believed to be
critical to producing a therapeutic effect in these disease
settings.
[0053] IRM compounds that are TLR 8 agonists may be particularly
desirable for use with therapeutic cancer vaccines because antigen
presenting cells that express TLR8 have been shown to produce IL-12
upon stimulation through TLR8. IL-12 is believed to play a
significant role in activation of CTLs, which are important for
mediating therapeutic efficacy as described above.
[0054] IRM compounds that are TLR 7 agonists and/or TLR 9 agonists
may be particularly desirable for use with prophylactic vaccines
because the type I interferon induced by stimulation through these
TLRs is believed to contribute to the formation of neutralizing
Th1-like humoral and cellular responses.
[0055] IRM compounds that are both TLR 7 and TLR 8 agonists may be
particularly desirable for use with therapeutic viral vaccines
and/or cancer vaccines because TLR7 stimulation is believed to
induce the production of type I IFN and activation of innate cells
such as macrophages and NK cells, and TLR8 stimulation is believed
to activate antigen presenting cells to initiate cellular adaptive
immunity as described above. These cell types are able to mediate
viral clearance and/or therapeutic growth inhibitory effects
against neoplasms.
[0056] IRM compounds that are non-TLR 7 agonists, and do not induce
substantial amounts of interferon alpha, may be desirable for use
with certain vaccines such as bacterial vaccines because TLR7
induces type I IFN production, which down-regulates the production
of IL-12 from macrophages and DCs. IL-12 contributes to the
subsequent activation of macrophages, NK cells and CTLs, all of
which contribute to anti-bacterial immunity. Therefore the
induction of anti-bacterial immunity against some kinds of bacteria
may be enhanced in the absence of IFNa.
[0057] For purposes of the present application, one way to
determine if an IRM compound is considered to be an agonist for a
particular TLR is if it activates an NFkB/luciferase reporter
construct through that TLR from the target species more than about
1.5 fold, and usually at least about 2 fold, in TLR transfected
host cells such as, e.g., HEK293 or Namalwa cells relative to
control transfectants. For information regarding TLR activation,
see, e.g., applications WO 03/043573, U.S. 60/447,179, U.S.
60/432,650, U.S. 60/432,651, and U.S. 60/450,484, WO 03/043588 and
the other IRM patents and applications disclosed herein.
[0058] Preferred IRM compounds include a 2-aminopyridine fused to a
five-membered nitrogen-containing heterocyclic ring.
[0059] Examples of classes of small molecule IRM compounds include,
but are not limited to, derivatives of imidazoquinoline amines
including but not limited to amide substituted imidazoquinoline
amines, sulfonamide substituted imidazoquinoline amines, urea
substituted imidazoquinoline amines, aryl ether substituted
imidazoquinoline amines, heterocyclic ether substituted
imidazoquinoline amines, amido ether substituted imidazoquinoline
amines, sulfonamido ether substituted imidazoquinoline amines, urea
substituted imidazoquinoline ethers, and thioether substituted
imidazoquinoline amines; tetrahydroimidazoquinoline amines
including but not limited to amide substituted
tetrahydroimidazoquinoline amines, sulfonamide substituted
tetrahydroimidazoquinoline amines, urea substituted
tetrahydroimidazoquinoline amines, aryl ether substituted
tetrahydroimidazoquinoline amines, heterocyclic ether substituted
tetrahydroimidazoquinoline amines, amido ether substituted
tetrahydroimidazoquinoline amines, sulfonamido ether substituted
tetrahydroimidazoquinoline amines, urea substituted
tetrahydroimidazoquinoline ethers, and thioether substituted
tetrahydroimidazoquinoline amines; imidazopyridine amines including
but not limited to amide substituted imidazopyridines, sulfonamido
substituted imidazopyridines, and urea substituted
imidazopyridines; 1,2-bridged imidazoquinoline amines; 6,7-fused
cycloalkylimidazopyridine amines; imidazonaphthyridine amines;
tetrahydroimidazonaphthyridine amines; oxazoloquinoline amines;
thiazoloquinoline amines; oxazolopyridine amines; thiazolopyridine
amines; oxazolonaphthyridine amines; and thiazolonaphthyridine
amines, such as those disclosed in, for example, U.S. Pat. Nos.
4,689,338; 4,929,624; 4,988,815; 5,037,986; 5,175,296; 5,238,944;
5,266,575; 5,268,376; 5,346,905; 5,352,784; 5,367,076; 5,389,640;
5,395,937; 5,446,153; 5,482,936; 5,693,811; 5,741,908; 5,756,747;
5,939,090; 6,039,969; 6,083,505; 6,110,929; 6,194,425; 6,245,776;
6,331,539; 6,376,669; 6,451,810; 6,525,064; 6,545,016; 6,545,017;
6,558,951; and 6,573,273; European Patent 0 394 026; U.S. Patent
Publication No. 2002/0055517; and International Patent Publication
Nos. WO 01/74343; WO 02/46188; WO 02/46189; WO 02/46190; WO
02/46191; WO 02/46192; WO 02/46193; WO 02/46749; WO 02/102377; WO
03/020889; WO 03/043572 and WO 03/045391.
[0060] Additional examples of small molecule IRMs said to induce
interferon (among other things), include purine derivatives (such
as those described in U.S. Pat. Nos. 6,376,501, and 6,028,076),
imidazoquinoline amide derivatives (such as those described in U.S.
Pat. No. 6,069,149), and benzimidazole derivatives (such as those
described in U.S. Pat. No. 6,387,938). 1H-imidazopyridine
derivatives (such as those described in U.S. Pat. No. 6,518,265)
are said to inhibit TNF and IL-1 cytokines. Other small molecule
IRMs said to be TLR 7 agonists are shown in U.S. 2003/0199461
A1.
[0061] Examples of small molecule IRMs that include a
4-aminopyrimidine fused to a five-membered nitrogen-containing
heterocyclic ring include adenine derivatives (such as those
described in U.S. Pat. Nos. 6,376,501; 6,028,076 and 6,329,381; and
in WO 02/08595).
[0062] In some applications, for example, the preferred IRM
compound is other than imiquimod or S-28463 (i.e., resiquimod:
4-Amino-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol).
[0063] Examples of particular IRM compounds include
2-propyl[1,3]thiazolo[4,5-c]quinolin-4-amine, which is considered
predominantly a TLR 8 agonist (and not a substantial TLR 7
agonist),
4-amino-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol,
which is considered predominantly a TLR 7 agonist (and not a
substantial TLR 8 agonist), and
4-amino-2-(ethoxymethyl)-.alpha.,.alpha.-dimethyl-6,7-
,8,9-tetrahydro-1H-imidazo[4,5-c]quinoline-1-ethanol, which is a
TLR 7 and TLR 8 agonist. In addition to its TLR 7 activity (and TLR
6 activity, but low TLR 8 activity),
4-amino-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]qu-
inoline-1-ethanol has beneficial characteristics, including that it
has a much lower CNS effect when delivered systemically compared to
imiquimod. Other examples of specific IRM compounds include, e.g.,
N-[4-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)butyl]-N'-c-
yclohexylurea,
2-methyl-1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyri-
din-4-amine,
1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amine- ,
N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimet-
hylethyl}methanesulfonamide,
N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinol-
in-1-yl)butyl]methanesulfonamide,
2-methyl-1-[5-(methylsulfonyl)pentyl]-1H-
-imidazo[4,5-c]quinolin-4-amine,
N-[4-(4-amino-2-propyl-1H-imidazo[4,5-c]q-
uinolin-1-yl)butyl]methanesulfonamide,
2-butyl-1-[3-(methylsulfonyl)propyl-
]-1H-imidazo[4,5-c]quinoline-4-amine,
2-butyl-1-{2-[(1-methylethyl)sulfony-
l]ethyl}-1H-imidazo[4,5-c]quinolin-4-amine,
N-{2-[4-amino-2-(ethoxymethyl)-
-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}-N'-cyclohexylurea,
N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimeth-
ylethyl} cyclohexanecarboxamide,
N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo-
[4,5-c]quinolin-1-yl]ethyl}-N'-isopropylurea. Resiquimod,
4-amino-2-ethoxymethyl-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinolin-
e-1-ethanol, may also be used in certain situations where a
combination TLR 7 and TLR 8 agonist is desired.
[0064] Other IRM compounds include large biological molecules such
as oligonucleotide sequences. Some IRM oligonucleotide sequences
contain cytosine-guanine dinucleotides (CpG) and are described, for
example, in U.S. Pat. Nos. 6,194,388; 6,207,646; 6,239,116;
6,339,068; and 6,406,705. Some CpG-containing oligonucleotides can
include synthetic immunomodulatory structural motifs such as those
described, for example, in U.S. Pat. Nos. 6,426,334 and 6,476,000.
CpG7909 is a specific example. Other IRM nucleotide sequences lack
CpG and are described, for example, in International Patent
Publication No. WO 00/75304. However, the large biological molecule
IRMs may be less susceptible to rapid clearance from a localized
tissue region and, consequently, the IRM depot preparations
described herein may be especially useful in connection with small
molecule IRMs described above.
[0065] Exemplary Applications:
[0066] IRM depot preparations delivered to a localized tissue
region can be used in a wide variety of applications, such as in
the treatment of a wide variety of conditions. For example, IRMs
such as imiquimod--a small molecule, imidazoquinoline IRM, marketed
as ALDARA (3M Pharmaceuticals, St. Paul, Minn.)--have been shown to
be useful for the therapeutic treatment of warts, as well as
certain cancerous or pre-cancerous lesions (See, e.g., Geisse et
al., J. Am. Acad. Dermatol., 47(3): 390-398 (2002); Shumack et al.,
Arch. Dermatol., 138: 1163-1171 (2002); U.S. Pat. No. 5,238,944 and
International Publication No. WO 03/045391.
[0067] Other diseases for which IRMs identified herein, including
as an IRM depot preparation, may be used as treatments include, but
are not limited to:
[0068] viral diseases, such as genital warts, common warts, plantar
warts, hepatitis B, hepatitis C, herpes simplex virus type I and
type II, molluscum contagiosum, variola, HIV, CMV, VZV, rhinovirus,
adenovirus, coronavirus, influenza, para-influenza;
[0069] bacterial diseases, such as tuberculosis, and mycobacterium
avium, leprosy;
[0070] other infectious diseases, such as fungal diseases,
chlamydia, candida, aspergillus, cryptococcal meningitis,
pneumocystis carnii, cryptosporidiosis, histoplasmosis,
toxoplasmosis, trypanosome infection, leishmaniasis;
[0071] neoplastic diseases, such as intraepithelial neoplasias,
cervical dysplasia, actinic keratosis, basal cell carcinoma,
squamous cell carcinoma, hairy cell leukemia, Karposi's sarcoma,
melanoma, renal cell carcinoma, myelogeous leukemia, multiple
myeloma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, and
other cancers;
[0072] TH-2 mediated, atopic, and autoimmune diseases, such as
atopic dermatitis or eczema, eosinophilia, asthma, allergy,
allergic rhinitis, systemic lupus erythematosis, essential
thrombocythaemia, multiple sclerosis, Ommen's syndrome, discoid
lupus, alopecia greata, inhibition of keloid formation and other
types of scarring, and enhancing would healing, including chronic
wounds; and
[0073] as a vaccine adjuvant for use in conjunction with any
material that raises either humoral and/or cell mediated immune
response, such live viral and bacterial immunogens and inactivated
viral, tumor-derived, protozoal, organism-derived, fungal, and
bacterial immunogens, toxoids, toxins, polysaccharides, proteins,
glycoproteins, peptides, cellular vaccines, DNA vaccines,
recombinant proteins, glycoproteins, and peptides, and the like,
for use in connection with, e.g., cancers, BCG, cholera, plague,
typhoid, hepatitis A, B, and C, influenza A and B, parainfluenza,
polio, rabies, measles, mumps, rubella, yellow fever, tetanus,
diphtheria, hemophilus influenza b, tuberculosis, meningococcal and
pneumococcal vaccines, adenovirus, HIV, chicken pox,
cytomegalovirus, dengue, feline leukemia, fowl plague, HSV-1 and
HSV-2, hog cholera, Japanese encephalitis, respiratory syncytial
virus, rotavirus, papilloma virus, and yellow fever.
[0074] The IRM depot preparations of the invention may be
particularly beneficial for use within solid tumors and cancerous
organs or tissue regions. If the residence time of the IRM is
extended within the cancerous tissue, it is believed that the
body's immune response to the cancer can be enhanced and directly
targeted to relevant tumor antigens. This not only may help reduce
or eliminate cancer at the site of IRM depot preparation delivery,
but, by sensitizing the immune system to the cancer, may help the
immune system attack the cancer in other locations throughout the
body. This approach to treatment may be used alone or in
conjunction with other treatments for the cancer, such as
therapeutic cancer vaccination (which may further include use of an
IRM depot preparation), antibody based therapies such as Rituxan
and Herceptin, and other chemotherapies. Examples of cancers that
may be particularly suitable for direct injection of an IRM depot
preparation into a localized tissue region include, but are not
limited to, breast cancer, lung cancer, stomach cancer, head and
neck cancer, colorectal cancer, renal cell carcinoma, pancreatic
cancer, basal cell carcinoma, cervical cancer, melanoma, prostate
cancer, ovarian cancer, and bladder cancer.
[0075] The methods, materials, and articles of the present
invention may be applicable for any suitable subject. Suitable
subjects include, but are not limited to, animals such as, but not
limited to, humans, non-human primates, rodents, dogs, cats,
horses, pigs, sheep, goats, cows, or birds. IRMs may also be
particularly helpful in individuals having compromised immune
functioning, such as those with HIV AIDS, transplant patients, and
cancer patients.
[0076] An amount of an IRM depot preparation effective for a given
therapeutic or prophylactic application is an amount sufficient to
achieve the intended therapeutic or prophylactic application. The
precise amount of IRM depot preparation used will vary according to
factors known in the art including but not limited to the physical
and chemical nature of the IRM compound, the nature of the
composition, the intended dosing regimen, the state of the
subject's immune system (e.g., suppressed, compromised,
stimulated), the method of administering the IRM compound, and the
species to which the formulation is being administered. Accordingly
it is not practical to set forth generally the amount that
constitutes an amount of IRM and IRM depot preparation effective
for all possible applications. Those of ordinary skill in the art,
however, can readily determine an appropriate amount with due
consideration of such factors.
EXAMPLE
[0077] The following example has been selected merely to further
illustrate features, advantages, and other details of the
invention. It is to be expressly understood, however, that while
the example serve this purpose, the particular materials and
amounts used as well as other conditions and details are not to be
construed in a matter that would unduly limit the scope of this
invention.
[0078] Mice were immunized via subcutaneous injection with 500 ug
ovalbumin protein alone (formulated as an aqueous solution in
phosphate buffered saline) or mixed with 200 ug of one of the
following IRMs: 1
[0079] Prepared as a non-depot preparation, formulated to deliver
200 ug as an aqueous solution in an acetate buffered saline mixture
containing cyclodextrin. 2
[0080] Prepared as a non-depot preparation, formulated to deliver
200 ug as an aqueous solution in a acetate buffered saline mixture
containing cyclodextrin. 3
[0081] IRM 3 is a lipidated IRM. IRM 3 was formulated to deliver
200 ug using a depot preparation formulated as a micropartical
precipitate (ppt), colloidal suspension, or as a micellar
suspension. The precipitate composition was a microparticle (>1
micron, around 10-500 microns) composition formed by simply
injecting IRM 3 dissolved in an organic solvent into an aqueous
solution without surfactants. The resulting particle size range was
broad, unlike the colloidal suspension (which provided a more
monomodal submicron size). To form the colloidal formulation, IRM 3
was first dissolved in an organic and water miscible solvent (e.g.
N-methylpyrrolidone, DMSO, Cremophore EL), then added to an aqueous
solution containing an appropriate amount of Tween 80. This causes
the lipophilic IRM 3 to precipitate into colloidal particles,
coated or surrounded by Tween 80 molecules, which act to prevent or
minimize flocculation or agglomeration of the IRM particles. The
higher the concentration of Tween 80, the finer the IRM colloidal
particle size will be. At a concentration of Tween 80 greater than
2%, the IRM colloidal suspension turned into a translucent, almost
clear solution, suggestive of a micellar encapsulation of the IRM
by Tween 80. In this approach, the IRM may also partition between
the inside of the micelle or be embedded in the micelle shell
itself, contributing to its stability.
[0082] IRM 4 is known as CpG 1826 and was formulated to deliver 200
ug as an aqueous solution in phosphate buffered saline.
[0083] In each of the above cases, mice were injected
subcutaneously 100 ul of formulation containing 200 ug of IRM and
500 ug of ovalbumin, then boosted 2 weeks later with same
formulation. Five days after the boost, the spleens were removed
and the cells stained with CD8, CD44, B220 and Kb-SIINFEKL
tetramer.
[0084] The results are shown in FIG. 1. The data shown was gated on
all live, CD8+, B220-events. The numbers in the upper right
quadrants indicate the percent tetramer staining cells (antigen
specific cells) out of total CD8+T cells. The lipidated IRM
molecule provides a clearly enhanced immune response than the non
lipidated molecules. It is believed that this is due to both the
lipid-modified nature of the compound, and the precipitate,
colloidal suspension, and micellar suspension compositions, causing
the IRM to remain resident within the localized tissue region for
an extended period.
[0085] The complete disclosures of the patents, patent documents
and publications cited herein are incorporated by reference in
their entirety as if each were individually incorporated. In case
of conflict, the present specification, including definitions,
shall control. Various modifications and alterations to this
invention will become apparent to those skilled in the art without
departing from the scope and spirit of this invention. Illustrative
embodiments and examples are provided as examples only and are not
intended to limit the scope of the present invention. The scope of
the invention is limited only by the claims set forth as
follows.
* * * * *
References