U.S. patent application number 16/438762 was filed with the patent office on 2019-12-19 for nanoparticles for crossing the blood brain barrier and methods of treatment using the same.
The applicant listed for this patent is California Institute of Technology. Invention is credited to Mark E. Davis, Emily A. Wyatt.
Application Number | 20190381188 16/438762 |
Document ID | / |
Family ID | 68839032 |
Filed Date | 2019-12-19 |
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United States Patent
Application |
20190381188 |
Kind Code |
A1 |
Davis; Mark E. ; et
al. |
December 19, 2019 |
Nanoparticles For Crossing The Blood Brain Barrier And Methods Of
Treatment Using The Same
Abstract
The present application discloses nanoparticles carrying
therapeutic agents, including chemotherapeutic agents, and
targeting ligands suitable for delivering these therapeutic agents
through the blood brain barrier and methods of using these patients
on those patients in need of such treatment.
Inventors: |
Davis; Mark E.; (Pasadena,
CA) ; Wyatt; Emily A.; (Encino, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
California Institute of Technology |
Pasadena |
CA |
US |
|
|
Family ID: |
68839032 |
Appl. No.: |
16/438762 |
Filed: |
June 12, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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62684593 |
Jun 13, 2018 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 31/4745 20130101;
A61K 9/5138 20130101; A61K 31/137 20130101; A61K 31/4045 20130101;
A61K 9/1641 20130101; B82Y 5/00 20130101; C07K 16/32 20130101; A61K
2039/505 20130101; A61P 35/00 20180101; A61K 47/6935 20170801 |
International
Class: |
A61K 47/69 20060101
A61K047/69; A61K 9/16 20060101 A61K009/16; A61K 31/4045 20060101
A61K031/4045; A61K 31/137 20060101 A61K031/137; A61K 31/4745
20060101 A61K031/4745; C07K 16/32 20060101 C07K016/32 |
Goverment Interests
GOVERNMENT RIGHTS
[0002] This invention was made with government support under Grant
No. CA151819 awarded by the National Institutes of Health. The
government has certain rights in the invention.
Claims
1. A method of treating a neurological disorder in a patient, the
method comprising systemically administering a first small molecule
therapeutic agent and/or a large molecule therapeutic agent to the
patient in need of such treatment, wherein (a) the first small
molecule therapeutic agent and/or the large molecule therapeutic
agent is attached to a nanoparticle comprising a nanoparticle core
and a targeting agent, the targeting agent comprising at least one
targeting ligand attached to an external surface of the
nanoparticle core by a linker having a cleavable linkage; (i) the
external surface of the nanoparticle comprising a neutral and/or
negatively charged mucic acid-containing polymer (MAP) (including
being substantially free of cationic mucic-acid containing polymer
(cMAP)); (ii) the at least one targeting ligand having an affinity
for binding to a receptor expressed by endothelial cells of the
blood-brain barrier; and (iii) the cleavable linkage being
cleavable when subject to conditions inside an endothelial cell of
the blood-brain barrier, wherein the cleavage comprises hydrolysis,
chemical reduction, or enzymatic cleavage of the linker; and
wherein one or both of (iv) the small molecule therapeutic agent is
optionally linked to the nanoparticle core by way of an optional
linker; and/or (v) the large molecule therapeutic agent is linked
to the nanoparticle by way of an optional linker; and (b) the
administration of the first small molecule therapeutic agent and/or
the large molecule therapeutic agent attached to the nanoparticle
results in the delivery of the first small molecule therapeutic
agent and/or the large molecule therapeutic agent past the blood
brain barrier and into the subject's brain parenchyma in an amount
is greater than would be delivered were the first small molecule
therapeutic agent and/or the large molecule therapeutic agent not
attached to the nanoparticle.
2. The method of claim 1, wherein the method comprises delivering
the large molecule therapeutic agent attached to the nanoparticle,
such that the large molecule therapeutic agent is delivered past
the blood brain barrier and into the subject's brain parenchyma in
an amount that is greater than would be delivered were the large
molecule therapeutic agent not attached to the nanoparticle.
3. The method of claim 2, wherein the amount of the large molecule
therapeutic agent that passes the blood brain barrier and into the
subject's brain parenchyma is a therapeutically effective amount
for the neurological disorder.
4. The method of claim 2, further comprising systemically
administering to the patient a second small molecule therapeutic
agent that itself able to pass the blood brain barrier and be
delivered into the subject's brain parenchyma in a therapeutically
effective amount, wherein the second small molecule therapeutic
agent is not attached to the nanoparticle.
5. The method of claim 4, wherein the first and second small
molecule therapeutic agents are not the same.
6. The method of claim 1, wherein the method comprises delivering
both the first small molecule therapeutic agent and the large
molecule therapeutic agent, both attached to the nanoparticle, such
that both the first small molecule therapeutic agent and the large
molecule therapeutic agent are delivered past the blood brain
barrier and into the subject's brain parenchyma in amounts that are
individually greater than would be delivered were the small
molecule therapeutic agent and the large molecule therapeutic agent
large molecule therapeutic agent not attached to the
nanoparticle.
7. The method of claim 6, wherein the amount of first small
molecule therapeutic agent and the large molecule therapeutic agent
that passes the blood brain barrier and into the subject's brain
parenchyma are individually a therapeutically effective amount for
the neurological disorder.
8. The method of claim 1, wherein the neurological disorder is
brain cancer.
9. The method of claim 1, wherein the nanoparticle core comprises a
polymer comprising units of the formula: ##STR00049##
10. The method of claim 1, wherein the nanoparticle core comprises
a polyol structure: ##STR00050## where y is in a range of from 10
to 25; where R is a functional group residue corresponding to that
of an amino acid containing a third functional group, for example
arginine (R is CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2),
histidine (R is CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof, and/or where
R is coupled to one or more of the targeting agent, the first small
molecule therapeutic agent, and/or the large molecule therapeutic
agent.
11. The method of claim 1, wherein the nanoparticle core comprises
a polymer derived from the coupling of a compound of Formula A with
a compound of Formula B; wherein the compound of Formula A is:
##STR00051## wherein n is a number from 1 to 20; and R is a
functional group residue (e.g., --COOH, --NH.sub.2, --OH)
corresponding to that of an amino acid containing a third
functional group, for example arginine (R is
CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2+), histidine (R is
CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof; and the
compound of Formula B is: ##STR00052## but not ##STR00053## in
which p is a number from 20 to 200; and L is a leaving group.
12. The method of claim 1, wherein the nanoparticle core comprises
a polymer comprises units of the formula: ##STR00054## wherein n is
a number from 1 to 20 and p is a number from 20 to 200.
13. The method of claim 1, wherein the cleavable linkage comprises
an acetal, a borate ester, a carbonate, a carboxylic acid ester, a
diamino ketal, a disulfide, a hydrazone, an imine, a ketal, an
orthoester, or a peptide linkage.
14. The method of claim 1, wherein the at least one targeting agent
comprises at least one borate ester of a (nitro)phenyl boronic
acid-containing polymer comprising a structure: ##STR00055##
wherein the nanoparticle core and the (nitro)phenyl boronic
acid-containing polymer are reversible connected to one another by
the borate condensation linkage between the (nitro)phenyl boronic
acid moieties of the (nitro)phenyl boronic acid-containing polymer
and at least one pair of vicinal diols of the nanoparticle core,
X.sub.5 being at the distal end of this connection; R.sup.A is
nitro; n is 1; s is a number in a range of from 2 to 2000; L is a
linking group between the phenyl ring and the polyethylene oxide
linkage, the linking group comprising an amide, carbonate, ester,
or disulfide group; and X.sub.5 is a C.sub.1-6 alkyl, optionally
substituted with --OH, --COOH, --B(OH).sub.2--,
--C(.dbd.O)O(alkyl), --C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl),
--N(alkyl).sub.2, or --SH, wherein the at least one targeting agent
is coupled thereto.
15. The method of claim 1, wherein the first small molecule
therapeutic agent is linked to the nanoparticle core by a linker
comprising an amino acid residue.
16. The method of claim 1, wherein the first small molecule
therapeutic agent is a neurotransmitter or a chemotherapeutic
agent.
17. The method of claim 15, wherein the first small molecule
therapeutic agent is dopamine, serotonin, camptothecin, irinotecan,
SN-38, or a metabolite, or prodrug thereof.
18. The method of claim 1, wherein the nanoparticle comprises a
unit structure: ##STR00056##
19. The method of claim 1, wherein the targeting ligand is
transferrin.
20. The method of claim 1, wherein the large molecule therapeutic
agent is trastuzumab (Herceptin).
21. The method of claim 1, wherein the first small molecule
therapeutic agent is camptothecin, irinotecan, SN-38, or a
metabolite, or prodrug thereof and the large molecule therapeutic
agent is trastuzumab (Herceptin).
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Patent Application
Ser. No. 62/684,593 filed Jun. 13, 2018, the contents of which are
incorporated by reference herein for all purposes.
TECHNICAL FIELD
[0003] The present disclosure is directed to the nanoparticles
capable of and for use in delivering therapeutic agents across the
blood brain barrier.
BACKGROUND
[0004] Chronic diseases of the central nervous system (CNS) are a
major cause of morbidity and mortality in the developed world.
Alzheimer's disease alone affects over five million people in the
United States and is expected to increase to over thirteen million
by 2050. Moreover, while the proportion of deaths from many other
leading causes of mortality in the United States, such as heart
disease and stroke, have seen significant decreases over the last
decade, the proportion of deaths from Alzheimer's disease has
increased 68%. A similar trend, in both high economic cost and a
relative lack of progress in treatment, is seen with many other
neurodegenerative diseases, including Huntington's disease,
Parkinson's disease, multiple sclerosis, and brain cancers.
[0005] Brain metastases of breast cancer are also presenting an
increasing challenge in the clinic. Historically, brain metastases
were not a major problem for most breast cancer patients because
they usually developed late during the disease, and lack of
systemic control limited survival. However, new therapies have
improved clinical outcomes in some subsets of patients, and brain
progression has become a more significant threat to long-term
survival. Certain of these cancers are caused by the overexpression
HER2, EGFR, HER3, and/or HER4 complexes. For example, in women with
human epidermal growth factor 2 (HER2)-positive breast cancer,
improved control of systemic disease with new therapies has
unmasked brain metastases that historically would have remained
clinically silent. Efficacy of therapeutic agents against brain
metastases is limited by their inability to permeate the
blood-brain and blood-tumor barriers (BBB and BTB) in therapeutic
amounts.
[0006] Although HER2-targeted therapies can effectively control
extracranial disease, they have limited distribution to brain
metastases and demonstrate poor efficacy in this setting. That is,
while HER2-targeted therapies effectively control systemic disease,
their efficacy against brain metastases is hindered by their
inability to penetrate the blood-brain and blood-tumor barriers
(BBB and BTB). Current therapeutic options such as surgery,
radiation and chemotherapy are considered palliative, and rarely
provide a significant increase in survival.
[0007] As with the delivery of most chemotherapeutics to the brain,
delivery of HER2-inhibitors to brain metastases is limited by poor
drug penetration across the blood-brain barrier (BBB), a selective
cellular barrier that acts as a regulator for the movement of
molecules into and out of the brain. The tumor microvasculature
associated with brain metastases, often referred to as the
blood-tumor barrier (BTB), has increased passive permeability
relative to the intact BBB; however, the loss in barrier integrity
is limited and highly variable from tumor to tumor and even within
the metastatic lesion. Many drugs commonly used to treat
HER2-positive breast cancer are unable to reach therapeutic
concentrations in the brain and circumventing the BBB and BTB
remains a major obstacle in effective treatment of brain
metastases.
[0008] A major reason for the lack of progress in treating these
diseases is due to the presence of the blood-brain barrier (BBB).
The BBB is a physical barrier between the CNS parenchyma and
vasculature that plays a critical role in maintaining homeostasis
within the CNS. The BBB consists of several barriers in parallel,
with the two that are best described being the vascular BBB,
consisting primarily of the capillary bed, and the
blood-cerebrospinal fluid (blood-CSF) barrier, consisting primarily
of the choroid plexus. Tight junctions exist between endothelial
cells that inhibit paracellular diffusion of polar molecules,
macromolecules and cells. This forces solute transport into the CNS
to occur primarily across individual endothelial cells. Though
critically important for maintaining CNS homeostasis, the
impermeability of the BBB to most solutes has proven a tremendous
obstacle for drug delivery to the CNS. Currently, 98% of small
molecule therapeutics and essentially 100% of large-molecule
therapeutics, including, monoclonal antibodies, proteins and gene
therapies, do not cross the BBB. Only a small class of drugs--small
molecules with high lipid solubility and a low molecular mass (Mr)
of <400-500 Daltons (Da)--actually cross the BBB by
themselves.
[0009] While a low molecular weight and high degree of lipid
solubility favor crossing by this mechanism, a drug taken up by the
membranes that form the BBB must then partition into the aqueous
environment of the brain's interstitial fluid to exert an effect.
As a result, a substance that is too lipid soluble can be
sequestered by the capillary bed of the barrier and not reach the
cells behind the BBB. Thus, while lipid solubility can increase
transport rate across the BBB, it can also lower the concentration
of the drug in the blood. The percent of administered drug entering
the brain is determined by both the rate of transport across the
BBB and the amount of drug presented to the brain. Use of lipid
solubility to improve drug delivery to the brain must thus find the
balance between increased permeation of the BBB and decreased
concentrations in blood.
[0010] There are only a few diseases of the brain that consistently
respond to this category of small molecules, and these include
depression, chronic pain, and epilepsy. In contrast, many other
serious illness disorders of the brain do not respond to the
conventional lipid-soluble-low-Mr small-molecule therapeutics, and
these include Alzheimer disease, stroke/neuroprotection, brain and
spinal cord injury, brain cancer, HIV infection of the brain,
various ataxia-producing disorders, amytrophic lateral sclerosis
(ALS), Huntington disease, Parkinson disease (PD), multiple
sclerosis (MS). and childhood inborn genetic errors affecting the
brain.
[0011] The present disclosure is directed to addressing some of
these challenges and concerns.
SUMMARY
[0012] The present disclosure is directed to nanoparticles suitable
for delivering therapeutics and imaging agents across the blood
brain barrier to treat a range of neurological conditions and
diseases that would benefit from the systemic delivery of these
therapeutics and imaging agents across the blood brain barrier.
Certain embodiments of the disclosure includes polymer, polymer
conjugate, or nanoparticle compositions comprising a polymer or
nanoparticle core to which is bonded: (a) at least one targeting
agent, the targeting agent comprising a ligand attached to an
external surface of the nanoparticle core by a linker; the ligand
having an affinity for binding to a receptor expressed by
endothelial cells of the blood-brain barrier; and the linker being
cleavable when subject to conditions inside an endothelial cell of
the blood-brain barrier, wherein the cleavage comprises hydrolysis,
chemical reduction, or enzymatic cleavage of the linker; and one or
both of (b) at least one small molecule therapeutic agent,
optionally linked to the nanoparticle core by way of an optional
linker; and/or (c) at least one large molecule therapeutic agent,
linked to the nanoparticle by way of an optional linker; wherein
the the large molecule therapeutic agent, when present, and the
targeting agent comprise different chemical entities. In certain
preferred embodiments, the composition is a nanoparticle comprises
the polymer or nanoparticle core to which is bonded at least one
targeting agent, the at least one small molecule therapeutic agent,
and the at least one large molecule therapeutic agent.
[0013] In other independent embodiments, the polymer or
nanoparticle core comprises, consists essentially of, or consists
of a polyol containing polymer, a poly(lactic-co-glycolic acid)
(PLGA), chitosan, polyethyleneimine, polysaccharide, polyester,
polyamide, polyether, polycarbonate, polyacrylate, iron oxide, or
gold. In such preferred embodiments, the the polymer or
nanoparticle core comprises, consists essentially of, or consists
of a polyol-containing polymer, preferably a sugar-containing
polymer, for example, a polymer derived from glucose, fructose,
mannitol, mucic acid, sucrose, galactose, sorbitol. xylose or
galactose, more preferably from mucic acid. A wide range of such
structures are set forth herein.
[0014] Additionally, or alternatively, the polymer or nanoparticle
core independently comprises units of the formula:
##STR00001##
[0015] Additionally, or alternatively, in some embodiments, the
polymer or nanoparticle core comprises a polymer containing a
polyol, wherein the polymer containing the polyol is derived from
the coupling of a compound of Formula A:
##STR00002##
[0016] with a compound of Formula B:
##STR00003##
wherein n, q, p, L, and R are described elsewhere herein.
Additionally, or alternatively, in some embodiments, the compound
of Formula B does not include:
##STR00004##
[0017] Additionally, or alternatively, in some embodiments, the
polymer or nanoparticle core comprises a polymer or nanoparticle
core polymer comprises a polyol structure:
##STR00005##
where X, Y, and R are defined elsewhere herein.
[0018] Additionally, or alternatively, in some embodiments, the
polymer or nanoparticle core comprises a polymer or nanoparticle
core polymer comprises alternating charged and uncharged segments
comprising one or more of the following structural units of Formula
(I) or Formula (II) or Formula (III):
##STR00006##
where A is an uncharged segment comprising polyalkylene glycol; and
B is a cationically charged segment comprising at least one
polyhydroxy linkage comprising at least one pair of vicinal diols.
In some of these embodiments, the B segment comprises at least one
repeating subunit comprising a caionic mucic acid polymer (cMAP)
residue structure. Various structures and permutations of these A
and B segments, and their associated structures, are set forth
elsewhere herein.
[0019] Additionally, or alternatively, in some independent
embodiments, the cleavable linker between the targeting ligand and
the polymer or nanoparticle core comprises one or more of an
acetal, borate ester, a carboxylic ester, a diaminoketal, a
disulfide, a ketal, a hydrazone, an imine, a ketal, an orthoester,
or a peptide linkage. These linkages are variously susceptible to
cleavage under the conditions associated with a brain endothelial
cell, for example by hydrolysis, chemical reduction, enzymatic
cleavage, or other means. In certain specific embodiments, the
cleavable linkage comprises at least one borate ester of a
(nitro)phenyl boronic acid-containing polymer comprising a
structure:
##STR00007##
where R.sup.A, n, S, and X.sup.5 are set forth elsewhere herein.
Various additional configurations of such linkages are set forth
herein. This linkage, cleavable within the endothelial cell, is
critical to these nanoparticles' ability to ferry cargo across the
blood brain barrier and other barriers.
[0020] In some embodiments the at least one targeting ligand
independently comprises or consists of a nucleotide,
polynucleotide, aptamer, peptide, oligopeptide, polypeptide,
protein, polysaccharide, antibody or antibody fragment.
Additionally, or alternatively, and as described elsewhere herein,
the at least one of the targeting ligands is one known to
specifically bind to receptor or surface protein expressed by a
brain endothelial cell that undergoes transcytosis. A wide variety
of useful targeting ligands are set forth elsewhere within this
disclosure. In certain additional or alternative embodiments
comprise or consist of, but are not limited to, chemical entities
that specifically bind to certain identified targeted receptors.
Transferrin is an attractive option for this targeting ligand and
serves as a specific embodiment in this capacity.
[0021] Each polymer or nanoparticle core can be conjugated to a
single targeting ligand (per core) or to several thousand such
targeting ligands (per core). Moreover, the the polymer or
nanoparticle core can be conjugated to a single type of targeting
ligand or multiple types. Further, while at least one targeting
ligand must be conjugated to each polymer or nanoparticle core by a
cleavable linkage, as set forth elsewhere herein, additional
targeting ligands can also be linked to the core with non-cleavable
linkages, offering added functionality to the structures. Still
further, the polymer or nanoparticle core may further comprise free
pendant moieties capable of acting as linking groups (cleavable or
otherwise) to which no targeting linkages are attached. Such
linkable groups offer the possibility for attaching other
biological, chemical, or imaging agents to the core, or may link
such biological, chemical, or imaging agents to the core.
[0022] In certain independent embodiments, the small molecule
optionally attached to the polymer or nanoparticle core is a
pharmaceutical compound useful in the treatment of Alzheimer's
disease, Huntington's disease, Parkinson's disease, multiple
sclerosis, and brain cancer. Such compounds include
neurotransmitters, chemotherapeutic, and other biologically active
materials. Compounds such as dopamine, serotonin, camptothecin,
irinotecan, SN-38, or a derivative, metabolite, or prodrug thereof
are but a small list of compounds set forth in this disclosure.
[0023] Alternatively, or additionally, the small molecule can also
be "tagged" with radio-isotopes for molecular imaging of the tumor
marker in vivo.
[0024] In certain independent additive or alternative embodiments,
the small molecule therapeutic compound may be chemically or
electrostatically bonded to the polymer or nanoparticle core or may
be encapsulated within the structure of the nanoparticle core
without being chemically or electrostatically bonded thereto. When
chemically linked, the small molecule linker may be either
chemically stable (i.e., able to maintain its structure in its
presented or intended environment) or may be cleavable by any of
the mechanisms associated with the term "cleavable" as set forth
elsewhere herein. Additionally, or alternatively, this linking
group may comprise one or more amino acid residues, such as a
residue of arginine, histidine, lysine, aspartic acid, glutamic
acid, serine, threonine, asparagine, glutamine, tyrosine,
tryptophan, or a salt thereof.
[0025] In certain independent embodiments, the large molecule
therapeutic agent is a nucleotide, polynucleotide, aptamer,
peptide, oligopeptide, polypeptide, protein (including fusion
proteins), polysaccharide, antibody or antibody fragment useful in
the treatment of Alzheimer's disease, Huntington's disease,
Parkinson's disease, multiple sclerosis, and cancer. In certain
Aspects of this Embodiment, the large molecule is an antibody. In
this regard, the present disclosure exemplifies the use of
trastuzumab (Herceptin.RTM.) in this capacity, though the large
molecule therapeutic agent is not so limited to this material, and
the disclosure sets for a wide range of materials useful in this
capacity.
[0026] As with the small molecule compound, the large molecule
therapeutic compound may be attached to the polymer or nanoparticle
core either by a direct chemical bond or via a linking group. The
linking group may be either chemically stable (i.e., able to
maintain its structure in its presented or intended environment) or
may be cleavable by any of the mechanisms associated with the term
"cleavable" as set forth elsewhere herein. Additionally, or
alternatively, this linking group may comprise one or more amino
acid residues, such as for example arginine, histidine, lysine,
aspartic acid, glutamic acid, serine, threonine, asparagine,
glutamine, tyrosine, tryptophan, or a salt thereof.
[0027] While the compositions have been disclosed in terms of a
polymer, polymer conjugate, or nanoparticle composition in
preferred embodiments, the composition is a nanoparticle. The
disclosure also embraces populations of nanoparticles, and
pharmaceutical compositions derived therefrom.
[0028] Additionally, the disclosure embraces methods of using these
nanoparticle compositions for systemically delivering enhanced
levels of therapeutics, including the combination of small and
large molecule therapeutics, past the blood brain barrier and into
the brain parenchyma. Such methods are directed to specific patient
patient populations, previously identified for such treatment.
These methods comprise systematically administering to such a
subject having a neurological brain disorder and in need of such
treatment the nanoparticles as disclosed herein. In this context,
the enhanced level of the therapeutic agent delivered by the
nanoparticles to the brain parenchyma can be defined as an amount
that is greater than is delivered using otherwise equivalent
nanoparticles that do not contain the cleavable linker under the
same conditions. Several means for determining the effectiveness
are set forth elsewhere herein.
[0029] In some embodiments, the disclosed treatments include those
where the attached small or large molecule therapeutic agent is
otherwise incapable of passing through the blood brain barrier to
deliver therapeutically effective amounts of the agent. In still
other embodiments, this administration is accompanied by the
co-administration of a therapeutic or imaging agent that is itself
able to pass the blood brain barrier and deliver therapeutic
amounts in the brain parenchyma. This tandem administration may be
provided at the same time, in the same delivery vehicle, or at
different times, in different delivery vehicles during a given
treatment regime. In this context, the enhanced level of the
therapeutic agent delivered by the nanoparticles to the brain
parenchyma can be defined as an amount that is greater than is
delivered using otherwise equivalent nanoparticles that do not
contain the cleavable linker under the same conditions.
[0030] The methods of administering these compositions, and the
certain neurological conditions suitable for treatment are set
forth elsewhere herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0032] The present application is further understood when read in
conjunction with the appended drawings. For illustrating the
subject matter, there are shown in the drawings exemplary
embodiments of the subject matter; however, the presently disclosed
subject matter is not limited to the specific methods, devices, and
systems disclosed. In addition, the drawings are not necessarily
drawn to scale. In the drawings:
[0033] FIG. 1(A) shows a scheme of acid-cleavable targeting
strategy. Following endocytosis, rapid acidification of endosome
triggers separation of Tf ligands from the nanoparticle core,
allowing free diffusion of the nanoparticle into the brain
parenchyma after transcytosis. FIG. 1(B) illustrates the
preparation of TfR-targeted and non-targeted MAP-CPT nanoparticles
and pH-dependence of nitroPBA-diol complex. x.about.82 for 3.4 kDa
PEG; y.about.20 for material used in this study; z.about.120 for 5
kDa PEG.
[0034] FIG. 2 illustrates a proposed mechanism for delivery of drug
and antibody combination to brain metastases using acid-cleavable
targeting ligands. At extracellular pH 7.4, Tf ligands and
Herceptin remain bound to the diols on the nanoparticle surface.
After endocytosis, rapid acidification of the endosome to pH 5.5
triggers their dissociation from the nanoparticle core, allowing
free diffusion into the brain once transcytosis is complete.
[0035] FIG. 3 shows a synthetic scheme of MAP-CPT polymer-drug
conjugate. w.about.82 for 3.4 kDa PEG; x.about.20 for material used
in this study.
[0036] FIG. 4(A-C). Single-agent or combination drug and antibody
nanoparticle delivery systems. Preparation of TfR-targeted
combination CPT/Herceptin nanoparticle (FIG. 4(A)), TfR-targeted
CPT nanoparticle (FIG. 4(B)) and TfR-targeted Herceptin
nanoparticle (FIG. 4(C)) formulations. w.about.82 for 3.4 kDa PEG;
x.about.20 for material used in this study; y.about.84 for 3.5 kDa
PEG; z.about.120 for 5 kDa PEG.
[0037] FIGS. 5(A-C) illustrates of breast cancer brain metastasis
models. FIGS. 5 (A): Intracranial (IC) injection of tumor cells
allows for direct establishment of brain metastases. FIG. 5 (B):
Following intracardiac (ICD) injection into the left ventricle,
tumor cells can head to brain vasculature, as well as to other
organs. Some cells will successfully extravasate and form
macroscopic brain tumors. FIG. 5 (C): After intravenous (IV)
injection, most tumor cells will arrest in the lung capillary bed,
as well as other sites, followed by subsequent metastasis to the
brain.
[0038] FIG. 6 shows apical to basal transport of non-targeted and
TfR-targeted MAP-CPT nanoparticles in model BBB. TfR-targeted
(blue) and non-targeted (gray) nanoparticles were added to apical
wells in either serum-free DMEM (DMEM), or in the presence of
either 2.5 mg/mL Tf (DMEM+Tf) or equimolar high-affinity anti-TfR
Ab (DMEM+Ab). Data shown are the average of 4 wells for each group.
Error bars indicate SE.
[0039] FIGS. 7(A-B) show the effect of anti-HER2 therapy on
HER2-positive BT474-Gluc breast cancer brain metastases established
in Rag2.sup.-/-;Il2rg.sup.-/- mice. Trastuzumab at 5 mg/kg was
administered twice weekly via intravenous tail vein injection, and
treatment was initiated when tumors reached 10 mm.sup.3 in volume.
MRI was used to monitor brain tumor size and response to treatment.
Tumors established by IC injection (FIG. 7(A)) showed significant
delay in tumor progression, whereas those established by IV
injection (FIG. 7(B)) did not.
[0040] FIG. 8 provides a detailed illustration of intracranial (A),
intracardiac (B), and intravenous (C) breast cancer brain
metastasis models and timelines for efficacy study. Numbers below
timeline indicate mean (range) time in months to establishment of
visible brain metastases (BM; .about.0.2 mm.sup.3 in volume) by
MRI. Thick arrows denote treatment schedule for the study, with 4
weekly doses administered once tumors reached .about.2 mm.sup.3 in
volume.
[0041] FIG. 9 shows data for brain tumors established using
different methods show differential response to therapeutics. Tumor
growth curves of BT474-Gluc metastatic brain tumors treated with
CPT (orange, 4 mg/kg), non-targeted MAP-CPT nanoparticles (gray, 4
mg CPT/kg), and TfR-targeted MAP-CPT nanoparticles (blue, 4 mg
CPT/kg) compared to saline (black) when established by IC (A), ICD
(B), and IV injection (C). Data shown are the average of 6 mice per
treatment group. Error bars indicate SE.
[0042] FIG. 10 shows blood Gluc activity of IC-established tumors
is correlated with tumor volume, as measured by MRI, for each
treatment group. Blood Gluc activity is plotted against tumor
volume for saline (black), CPT (orange), non-targeted MAP-CPT
nanoparticle (gray), and TfR-targeted MAP-CPT nanoparticle (blue)
treatment groups. Linear regression was performed using MATLAB.
[0043] FIGS. 11(A-C) show individual tumor growth curves of
BT474-Gluc metastatic brain tumors treated with CPT (orange, 4
mg/kg), non-targeted MAP-CPT nanoparticles (gray, 4 mg CPT/kg), and
TfR-targeted MAP-CPT nanoparticles (blue, 4 mg CPT/kg) compared to
saline (black) when established by IC (FIG. 11(A)), ICD (FIG.
11(B)), and IV injection (FIG. 11(C)). Open circles denote
leptomeningeal metastases. 2 of 24 total metastases included in
this study for both the ICD and IV models were leptomeningeal. All
other brain tumors were intracerebral.
[0044] FIGS. 12(A-B) show brain uptake of therapeutics is
model-dependent in tumor, but not healthy tissue. FIG. 12(A) shows
brain uptake in BT474-Gluc tumor tissue as calculated by percent
injected dose per g of tissue for different treatments. FIG. 12(B)
Percent injected dose in healthy brain tissue. Brain uptake was
determined 24 h after a 4 mg/kg dose (CPT basis). Data shown are
the average of 4 mice per treatment group. Error bars indicate SE.
ND, not detectable.
[0045] FIG. 13 shows BT474-Gluc cells isolated from brain tumors
following IC-(circle), ICD- (square), and IV-establishment
(triangle) as well as parental cells (solid circle) are similarly
sensitive to CPT. Data shown are the average of 4 dose-response
curves for each cell line. Error bars indicate SE.
[0046] FIG. 14(A-B) shows synthetic schemes for nitroPBA
conjugates. (FIG. 14(A)) Herceptin-PEG3.5 k-nitroPBA. y.about.84
for 3.5 kDa PEG. (FIG. 14(B)) Tf-PEG5 k-nitroPBA. z.about.120 for 5
kDa PEG.
[0047] FIG. 15 shows schemes for synthesis of MAP-AF568 polymer
conjugate. w.about.82 for 3.4 kDa PEG; x.about.20 for material used
in Examples.
[0048] FIG. 16 shows representative MRI images of metastatic brain
tumors at the start of treatment (A), and 8 weeks after the start
of treatment with saline (B), free CPT and Herceptin (C),
TfR-targeted CPT nanoparticles (D), TfR-targeted Herceptin
nanoparticles (E) and TfR-targeted combination CPT/Herceptin
nanoparticles (F). Dotted lines denote tumor margins. Formulations
were systemically administered weekly for 4 weeks at a dose of 4
and/or 24 mg/kg (CPT and/or Herceptin bases, respectively). Scale
bar, 2 mm; NP, nanoparticle.
[0049] FIG. 17 shows images of metastatic brain tumors imaged by
MRI following IV injection of BT474-Gluc cells. Intracerebral
(A,B,C) and leptomeningeal metastases were detected (D). Most
intracerebral metastases were in the cerebrum (A), with occasional
metastases in the cerebellum (B). Multifocal metastases were
occasionally observed (C). Leptomeningeal metastases most commonly
grew in the subarachnoid space (D). Scale bar, 2 mm.
[0050] FIG. 18. Combined nanoparticle delivery of CPT and Herceptin
inhibits brain metastatic tumor growth more effectively than
nanoparticle delivery of either monotherapy and combined free drug.
Tumor growth curves of BT474-Gluc metastatic brain tumors treated
with free CPT and Herceptin (gray, 4 and 24 mg/kg, respectively),
TfR-targeted CPT nanoparticles (orange, 4 mg CPT/kg), TfR-targeted
Herceptin nanoparticles (purple, 24 mg Herceptin/kg) and
TfR-targeted combination CPT/Herceptin nanoparticles (blue, 4 mg
CPT/kg and 24 mg Herceptin/kg) compared to saline (black). Data
shown are the average of 6 mice per treatment group. Error bars
indicate SE.
[0051] FIG. 19 shows individual tumor growth curves of BT474-Gluc
metastatic brain tumors treated with CPT and Herceptin (gray, 4 and
24 mg/kg, respectively), TfR-targeted CPT nanoparticles (orange, 4
mg CPT/kg), TfR-targeted Herceptin nanoparticles (purple, 24 mg
Herceptin/kg) and TfR-targeted combination CPT/Herceptin
nanoparticles (blue, 4 mg CPT/kg and 24 mg Herceptin/kg) compared
to saline (black). NP, nanoparticle.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
[0052] The present disclosure is directed to nanoparticles
comprising targeting agents and therapeutics and/or imaging agents,
suitable for, but not limited to, delivering the therapeutics
and/or imaging agents to the brain of a subject. In some
embodiments, the the described nanoparticles are loaded with the
therapeutic agent and/or imaging agent of interest prior to
administration of the nanoparticle to the subject. Following
delivery of the loaded nanoparticle, the targeting ligand
facilitates delivery of the nanoparticle to a target cell of
interest, such as a brain endothelial cell. Following
internalization by the target cell, the nanoparticle dissociates
from the targeting agent. In the case of brain endothelial cells,
the internalized nanoparticle will then be excreted from the cell
into the interstitial space of the brain where the particle will
destabilize and secrete the loaded agent, thereby delivering the
agent to the brain or other target location. In some embodiments
the described methods may be carried out to deliver a
neurotransmitter such as serotonin or dopamine to the brain, which
may be used to treat a neurological disorder. Other agents for use
in treating neurological disorders may also be delivered to the
brain via the described methods. Imaging agents that might not
readily access the brain on their own may also be delivered using
the described methods. In some embodiments the described methods
may be used to deliver a nanoparticle carrying the imaging agent
Cu64 to the brain of a subject to allow for imaging. Further, the
described methods may be used to deliver a combination of one or
more therapeutic agents, imaging agents, or both therapeutic
agents, imaging agent to the brain of a subject.
[0053] In certain embodiments, the nanoparticles contain
therapeutics suitable for the control and treatment of
cancer-tumors, especially cancer-tumors of the brain. More
specifically, the instant disclosure materials and methods which
allow therapeutics to cross the blood-brain-barrier (BBB) and
blood-tumor-barriers (BTB) for the treatment of conditions
including metastatic, HER2-positive brain tumors.
[0054] There has been significant interest in engineering
nanoparticles and other nanoscale or polymeric drug formulations to
enhance the delivery of therapeutic agents to the brain following
systemic administration. The use of endogenous transport mechanisms
at the BBB such as receptor-mediated transcytosis (RMT) has emerged
as a promising approach to shuttle a variety of payloads across the
BBB. In particular, transferrin receptor (TfR) has been one of the
primary targets investigated for RMT because of its high expression
on the blood side of the BBB endothelium.
[0055] As described herein, the inventors investigated the brain
uptake and efficacy of TfR-targeted therapeutic nanoparticles
designed to transcytose the BBB/BTB. Transferrin (Tf) was attached
to nanoparticles consisting of a mucic acid polymer (MAP) conjugate
of camptothecin (CPT), denoted MAP-CPT, through a pH-dependent,
boronic acid-diol complexation to the vicinal diols contained
within the mucic acid portions of the polymer. With this
acid-cleavable targeting strategy, nanoparticles retain high
avidity to TfR on the blood side of the BBB to enable practical,
systemic dosing, yet release the targeting agents upon
acidification during transcytosis to allow their release into the
brain. FIGS. 1(A-B). The present inventors have demonstrated that
these targeted nanoparticles, administered systemically, were able
to deliver CPT to HER2-positive breast cancer brain metastases in
mice and eliciting a considerable antitumor response.
[0056] The present inventors further hypothesized that TfR-targeted
nanoparticles carrying more potent therapeutic agents would reveal
even greater tumor size reductions. In this disclosure, an
exemplary delivery system was investigated for its ability to
shuttle an anti-HER2 monoclonal antibody, Herceptin, alone or in
combination with a CPT payload, across the BBB to achieve enhanced
antitumor activity over the previously reported efficacy of CPT
alone (FIG. 2).
[0057] The present application discloses nanoparticles carrying
therapeutic agents, including chemotherapeutic agent, and targeting
ligands, where the nanoparticles are suitable for delivering these
chemotherapeutic agents throughout a patient, including through the
blood the blood brain barrier and methods of using these patients
on those patients in need of such treatment. While described in
terms of chemotherapy, it is appreciated that some of the
nanoparticles are also capable of carrying therapeutics not
directed to cancer across the blood brain barrier for treatment of
other neurological conditions, for example serotonin or dopanine
for treatment of diseases such as Parkinson's disease, Huntington
disease, and multiple sclerosis, such as described in U.S. Patent
Application Publ. No. 2014/0348754, which is incorporated by
reference herein for all purposes, but at least for the use of the
nanoparticles for this purpose.
[0058] The present disclosure in its full context, includes
sections labeled General Terms, General Embodiments of the Present
Disclosure, and Examples. These sections more fully describe the
following enumerated aspects of the present disclosure, as well as
embodiments not listed as follows. None of these sections, or
provisions that follow, are to be interpreted as limiting the
disclosure to any particular section, and should be considered in
total for the teachings of the present disclosure.
[0059] ASPECT 1: A method of treating a neurological disorder in a
patient, the method comprising systemically administering a first
small molecule therapeutic agent and/or a large molecule
therapeutic agent to the patient in need of such treatment,
wherein
[0060] (a) the first small molecule therapeutic agent and/or the
large molecule therapeutic agent is attached to a nanoparticle
comprising a nanoparticle core and a targeting agent, the targeting
agent comprising at least one targeting ligand attached to an
external surface of the nanoparticle core by a linker having a
cleavable linkage; [0061] (i) the external surface of the
nanoparticle comprising a neutral and/or negatively charged mucic
acid-containing polymer (MAP) (including where the nanoparticle
core and/or surface is substantially free of cationic mucic-acid
containing polymer (cMAP)); [0062] (ii) the at least one targeting
ligand having an affinity for binding to a receptor expressed by
endothelial cells of the blood-brain barrier; and [0063] (iii) the
cleavable linkage being cleavable when subject to conditions inside
an endothelial cell of the blood-brain barrier, wherein the
cleavage comprises hydrolysis, chemical reduction, or enzymatic
cleavage of the linker; and wherein one or both of [0064] (iv) the
small molecule therapeutic agent is optionally linked to the
nanoparticle core by way of an optional linker; and/or [0065] (v)
the large molecule therapeutic agent is linked to the nanoparticle
by way of an optional linker; and
[0066] (b) the administration of the first small molecule
therapeutic agent and/or the large molecule therapeutic agent
attached to the nanoparticle results in the delivery of the first
small molecule therapeutic agent and/or the large molecule
therapeutic agent past the blood brain barrier and into the
subject's brain parenchyma in an amount is greater than would be
delivered were the first small molecule therapeutic agent and/or
the large molecule therapeutic agent not attached to the
nanoparticle.
[0067] ASPECT 2. The method of ASPECT 1, wherein the method
comprises delivering the large molecule therapeutic agent attached
to the nanoparticle, such that the large molecule therapeutic agent
is delivered past the blood brain barrier and into the subject's
brain parenchyma in an amount that is greater than would be
delivered were the large molecule therapeutic agent not attached to
the nanoparticle.
[0068] ASPECT 3. The method of ASPECT 1 or 2, wherein the amount of
the large molecule therapeutic agent that passes the blood brain
barrier and into the subject's brain parenchyma is a
therapeutically effective amount for the neurological disorder.
[0069] ASPECT 4. The method of any one of ASPECTS 1 to 3, further
comprising systemically administering to the patient a second small
molecule therapeutic agent that itself able to pass the blood brain
barrier and be delivered into the subject's brain parenchyma in a
therapeutically effective amount, wherein the second small molecule
therapeutic agent is not attached to the nanoparticle.
[0070] ASPECT 5. The method of any one of ASPECTS 1 to 4, wherein
the first and second small molecule therapeutic agents are not the
same.
[0071] ASPECT 6. The method of any one of ASPECTS 1 to 5, wherein
the method comprises delivering both the first small molecule
therapeutic agent and the large molecule therapeutic agent, both
attached to the nanoparticle, such that both the first small
molecule therapeutic agent and the large molecule therapeutic agent
are delivered past the blood brain barrier and into the subject's
brain parenchyma in amounts that are individually greater than
would be delivered were the small molecule therapeutic agent and
the large molecule therapeutic agent large molecule therapeutic
agent not attached to the nanoparticle.
[0072] ASPECT 7. The method of any one of ASPECTS 1 to 6, wherein
the amount of first small molecule therapeutic agent and the large
molecule therapeutic agent that passes the blood brain barrier and
into the subject's brain parenchyma are individually a
therapeutically effective amount for the neurological disorder.
[0073] ASPECT 8. The method of any one of ASPECTS 1 to 7, wherein
the neurological disorder is brain cancer, including brain cancer
metastasized other systemic, extracranial cancers, including brain
cancer metastasized from HER2-positive breast cancer.
[0074] ASPECT 9. The method of any one of ASPECTS 1 to 8, wherein
the nanoparticle core comprises a polymer comprising units of the
formula:
##STR00008##
[0075] ASPECT 10. The method of any one of ASPECTS 1 to 9, wherein
the nanoparticle core comprises a polyol structure:
##STR00009##
where y is in a range of from 10 to 25;
[0076] where R is a functional group residue corresponding to that
of an amino acid containing a third functional group, for example
arginine (R is CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2.sup.+),
histidine (R is CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof, and/or where
R is coupled to one or more of the targeting agent, the first small
molecule therapeutic agent, and/or the large molecule therapeutic
agent.
[0077] ASPECT 11. The method of any one of ASPECTS 1 to 10, wherein
the nanoparticle core comprises a polymer derived from the coupling
of a compound of Formula A with a compound of Formula B; wherein
the compound of Formula A is:
##STR00010##
wherein n is a number from 1 to 20; and
[0078] R is a functional group residue (e.g., --COOH, --NH.sub.2,
--OH) corresponding to that of an amino acid containing a third
functional group, for example arginine (R is
CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2+), histidine (R is
CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof; and
[0079] the compound of Formula B is:
##STR00011##
but not
##STR00012##
in which
[0080] p is a number from 20 to 200; and
[0081] L is a leaving group.
[0082] ASPECT 12. The method of any one of ASPECTS 1 to 11, wherein
the nanoparticle core comprises a polymer comprises units of the
formula:
##STR00013##
wherein n is a number from 1 to 20 and p is a number from 20 to
200.
[0083] ASPECT 13. The method of any one of ASPECTS 1 to 13, wherein
the cleavable linkage comprises an acetal, a borate ester, a
carbonate, a carboxylic acid ester, a diamino ketal, a disulfide, a
hydrazone, an imine, a ketal, an orthoester, or a peptide
linkage.
[0084] ASPECT 14. The method of any one of ASPECTS 1 to 13, wherein
the at least one targeting agent comprises at least one borate
ester of a (nitro)phenyl boronic acid-containing polymer comprising
a structure:
##STR00014##
wherein
[0085] the nanoparticle core and the (nitro)phenyl boronic
acid-containing polymer are reversible connected to one another by
the borate condensation linkage between the (nitro)phenyl boronic
acid moieties of the (nitro)phenyl boronic acid-containing polymer
and at least one pair of vicinal diols of the nanoparticle core,
X.sub.5 being at the distal end of this connection;
[0086] R.sup.A is nitro;
[0087] n is 1;
[0088] s is a number in a range of from 2 to 2000;
[0089] L is a linking group between the phenyl ring and the
polyethylene oxide linkage, the linking group comprising an amide,
carbonate, ester, or disulfide group; and
[0090] X.sub.5 is a C.sub.1-6 alkyl, optionally substituted with
--OH, --COOH, --B(OH).sub.2--, --C(.dbd.O)O(alkyl),
--C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl), --N(alkyl).sub.2, or
--SH, wherein the at least one targeting agent is coupled
thereto.
[0091] ASPECT 15. The method of any one of ASPECTS 1 to 14, wherein
the first small molecule therapeutic agent is linked to the
nanoparticle core by a linker comprising an amino acid residue.
[0092] ASPECT 16. The method of any one of ASPECTS 1 to 15, wherein
the first small molecule therapeutic agent is a neurotransmitter or
a chemotherapeutic agent.
[0093] ASPECT 17. The method of any one of ASPECTS 1 to 16, wherein
the first small molecule therapeutic agent is dopamine, serotonin,
camptothecin, irinotecan, SN-38, or a metabolite, or prodrug
thereof.
[0094] ASPECT 18. The method of any one of ASPECTS 1 to 17, wherein
the nanoparticle comprises a unit structure:
##STR00015##
[0095] ASPECT 19. The method of any one of ASPECTS 1 to 18, wherein
the targeting ligand is transferrin.
[0096] ASPECT 20. The method of any one of ASPECTS 1 to 19, wherein
the large molecule therapeutic agent is trastuzumab
(Herceptin).
[0097] ASPECT 21. The method of any one of ASPECTS 1 to 20, wherein
the first small molecule therapeutic agent is camptothecin,
irinotecan, SN-38, or a metabolite, or prodrug thereof and the
large molecule therapeutic agent is trastuzumab (Herceptin).
General Terms
[0098] In the present disclosure the singular forms "a," "an," and
"the" include the plural reference, and reference to a particular
numerical value includes at least that particular value, unless the
context clearly indicates otherwise. Thus, for example, a reference
to "a material" is a reference to at least one of such materials
and equivalents thereof known to those skilled in the art.
[0099] When a value is expressed as an approximation by use of the
descriptor "about," it will be understood that the particular value
forms another embodiment. In general, use of the term "about"
indicates approximations that can vary depending on the desired
properties sought to be obtained by the disclosed subject matter
and is to be interpreted in the specific context in which it is
used, based on its function. The person skilled in the art will be
able to interpret this as a matter of routine. In some cases, the
number of significant figures used for a particular value may be
one non-limiting method of determining the extent of the word
"about." In other cases, the gradations used in a series of values
may be used to determine the intended range available to the term
"about" for each value. Where present, all ranges are inclusive and
combinable. That is, references to values stated in ranges include
every value within that range.
[0100] It is to be appreciated that certain features of the
invention which are, for clarity, described herein in the context
of separate embodiments, may also be provided in combination in a
single embodiment. That is, unless obviously incompatible or
specifically excluded, each individual embodiment is deemed to be
combinable with any other embodiment(s) and such a combination is
considered to be another embodiment. Conversely, various features
of the invention that are, for brevity, described in the context of
a single embodiment, aspects, Embodiment, or Aspect may also be
provided separately or in any sub-combination. Finally, while an
embodiment may be described as part of a series of steps or part of
a more general structure, each said step may also be considered an
independent embodiment in itself, combinable with others. Still
further, where certain features are described in terms of a one
category (e.g., in terms of a method or composition), it is to be
understood that that feature is equally applicable to all
categories (e.g., features described in the context of compositions
are also applicable to methods, and vice versa).
[0101] The transitional terms "comprising," "consisting essentially
of," and "consisting" are intended to connote their generally in
accepted meanings in the patent vernacular; that is, (i)
"comprising," which is synonymous with "including," "containing,"
or "characterized by," is inclusive or open-ended and does not
exclude additional, unrecited elements or method steps; (ii)
"consisting of" excludes any element, step, or ingredient not
specified in the claim; and (iii) "consisting essentially of"
limits the scope of a claim to the specified materials or steps and
those that do not materially affect the basic and novel
characteristic(s) of the claimed invention. Embodiments described
in terms of the phrase "comprising" (or its equivalents), also
provide, as embodiments, those which are independently described in
terms of "consisting of" and "consisting essentially of." For those
composition embodiments provided in terms of "consisting
essentially of," the basic and novel characteristic(s) is the
ability to provide the described effect associated with the
description as described herein or as explicitly specified. In
particular here, the basic and novel characteristic of the methods
and compositions are the ability, at least, to deliver cargo across
the blood brain barrier using systemic administrations at levels
that would not be achieved by the systemic administration of the
cargo by itself.
[0102] When a list is presented, unless stated otherwise, it is to
be understood that each individual element of that list, and every
combination of that list, is a separate embodiment. For example, a
list of embodiments presented as "A, B, or C" is to be interpreted
as including the embodiments, "A," "B," "C," "A or B," "A or C," "B
or C," or "A, B, or C." That is, where lists are provided within an
embodiment, additional embodiments include those lists that exclude
one or more elements of the first list, without the need for
specific exclusions. For example, embodiments described in terms of
the amino acid residues arginine, histidine, lysine, aspartic acid,
glutamic acid, serine, threonine, asparagine, glutamine, tyrosine,
tryptophan, or a salt thereof also include, as separate
embodiments, lists where one or more of these amino acids are
excluded
[0103] Throughout this specification, words are to be afforded
their normal meaning, as would be understood by those skilled in
the relevant art.
[0104] Where a composition is administered to a patient or subject,
the term "administration" means application of the composition to
the patient or subject. The terms "systemic administration,"
"administered systemically," "peripheral administration" and
"administered peripherally" as used herein mean the administration
of a nanoparticle or composition thereof other than directly into
the central nervous system, such that it enters the individual's
system and, thus, is subject to metabolism and other like
processes, for example, subcutaneous administration. The terms
"parenteral administration" and "administered parenterally" as used
herein means modes of administration other than enteral and topical
administration, usually by injection, and includes, without
limitation, intravenous, intramuscular, intra-arterial,
intrathecal, intracapsular, intraorbital, intracardiac,
intradermal, intraperitoneal, transtracheal, subcutaneous,
subcuticular, intraarticular, subcapsular, subarachnoid, and
intrastemal, injection and infusion.
[0105] Formulations suitable for parenteral administration usually
comprise a sterile aqueous preparation of the active compound,
which preferably is isotonic with the blood of the recipient (e.g.,
physiological saline solution). Such formulations may be presented
in unit-dose or multi-dose form.
[0106] Actual dosage levels of the active ingredient or agent in
the pharmaceutical compositions herein described may be varied to
obtain an amount of the active ingredient which is effective to
achieve the desired therapeutic response for a particular
individual, composition, and mode of administration, without being
toxic to the individual.
[0107] In some embodiments, the patients or subjects are
administered at least one of the described nanoparticles or
populations of nanoparticles in an individual or daily dose in a
range of from about 0.01 .mu.g to 5000 mg per dose or per kg of the
weight of the subject. In additional or alternative embodiments,
the patient or subject is administered at least one of the
nanoparticles or populations of nanoparticles set forth herein at a
daily or individual dose in a range of from 0.01 .mu.g to 0.1
.mu.g, from 0.1 .mu.g to 1 .mu.g, from 1 .mu.g to 10 .mu.g, from 10
.mu.g to 100 .mu.g, from 100 .mu.g to 1000 .mu.g, from 1 mg to 5
mg, from 5 mg to 10 mg, from 10 mg to 50 mg, from 50 mg to 100 mg,
from 100 mg to 150 mg, from 150 mg to 200 mg, from 200 mg to 250
mg, from 250 mg to 500 mg, from 500 mg to 750 mg, from 750 mg to
1000 mg, from 1000 mg to 2000 mg, from 2000 mg to 3000 mg, from
3000 mg to 4000 mg, or from 4000 mg to 5000 mg per dose or per kg
of the weight of the subject, or the daily or individual dose dose
is defined by two or more of the foregoing ranges, for example,
from 0.1 .mu.g to 1 mg per kg of body weight or from 1 mg to 10 mg
per dose or from 10 mg to 1000 mg per day. In some embodiments, the
described methods may be carried out so the nanoparticles described
herein is administered to a subject weekly, bi-weekly, monthly,
bi-month, semiannually, or annually. Treatment may be initiated
with smaller dosages that are less than the optimum dose followed
by an increase in dosage over the course of the treatment until the
optimum effect under the circumstances is reached.
[0108] The term "agent" as used herein indicates a compound capable
of exhibiting a chemical or biological activity associated to the
target. The term "chemical activity" as used herein indicates the
ability of the molecule to perform a chemical reaction. The term
biological activity as used herein indicates the ability of the
molecule to affect a living matter. Exemplary chemical activities
of agents comprise formation of a covalent or electrostatic
interaction. Exemplary biological activities of agents comprise
production and secretion of endogenous molecules, absorption and
metabolization of endogenous or exogenous molecules and activation
or deactivation of genetic expression including transcription and
translation of a gene of interest.
[0109] The term "antibody" includes reference to an immunoglobulin
molecule that is reactive with a particular antigen. The term also
includes genetically engineered forms such as chimeric antibodies
(e.g., humanized murine antibodies), heteroconjugate antibodies
(e.g., bispecific antibodies) and recombinant single chain Fv
fragments (scFv), disulfide stabilized (dsFv) Fv fragments, or pFv
fragments. The term "antibody" also includes antigen binding forms
of antibodies (e.g., Fab', F(ab').sub.2, Fab, Fv and rIgG). In the
present disclosure, trastuzumab (Herceptin.RTM.) is a useful
antibody, and a preferred embodiment, for the treatment of breast
cancer, and for cancers that are HER2 positive.
[0110] An antibody immunologically reactive with, or "specific
for," a particular antigen is a relative term and means that the
antibody binds to that antigen with an affinity that is at least
10-old higher than would be observed for non-specific binding
exhibited by the antibody. Thus, an antibody said to be "specific
for" a given antigen may in fact selectively bind other antigens
with an affinity that is 10-fold-high than it exhibits in
nonspecific interactions.
[0111] The term "attach", "attached" or "attachment" as used
herein, refers to connecting or uniting by a bond, link, force or
tie in order to keep two or more components together, which
encompasses either direct or indirect attachment such that for
example where a first compound is directly bound to a second
compound, and the embodiments wherein one or more intermediate
compounds, and in particular molecules, are disposed between the
first compound and the second compound.
[0112] Where used herein, the term "cargo" refers to the
therapeutic or imaging agents being delivered (as if by "ferry") by
the nanoparticles described herein
[0113] In the nanoparticles herein described polyols polymers are
coupled to the boronic acid polymers. The term "coupled" or
"coupling" as used herein with reference to attachment between two
molecules indicates an interaction forming a reversible covalent
linkage. In the presence of a suitable medium, a boronic acid
presented on the boronic acid polymer interact with hydroxyl groups
of the polyols associated with the nanoparticle core via a rapid
and reversible pair-wise covalent interaction to form boronic
esters in a suitable medium. Typically, the boronic acid moieties
are suitable for forming borate esters with the hydroxy groups of
the nanoparticle surfaces, and for hydrolyzing under certain
triggering events (e.g., changes in pH). Suitable media include
water and several aqueous solutions and additional organic media
identifiable by a skilled person. Typically, when contacted in an
aqueous medium boronic acid polymers and polyols polymers react in
a condensation reaction producing water as a side product. The
boronic acid polyol interaction is generally more favorable in
aqueous solutions but is also known to proceed in organic media. In
addition, cyclic esters formed with 1,2 and 1,3 diols are generally
more stable than their acyclic ester counterparts.
[0114] The term "brain cancer" refers to a condition where the
cancer cells are present in the brain of the patient or subject,
typically arising from metastases from other cancers outside the
brain ("extracranial cancers"). When the brain cancer arises from
the metastates of other extracranial cancers (for example, breast
cancer), the methods and compositions disclosed herein allow for
the use of those therapeutic agents, suitable for treating these
extracranial cancers, but which are otherwise unable to pass the
BBB, to deliver therapeutic levels of the metastasized brain
cancers.
[0115] The term "conjugated" as used herein indicates that one
molecule has fobrmed a covalent bond with a second molecule. The
term also refers to linkages where atoms covalently bond with
alternating single and multiple (e.g. double) bonds (e.g.,
C.dbd.C--C.dbd.C--C) and influence each other to produce electron
delocalization. The person of skill in the art would be able to
understand and distinguish these meanings, depending on the
context.
[0116] The terms "deliver" and "delivery," as used herein, indicate
the activity of affecting the spatial location of a compound, and,
in particular, specifying the preferred location of a compound.
Accordingly, delivering a compound in the sense of the present
disclosure indicates the ability to affect positioning and movement
of the compound at a certain time under a certain set of
conditions, so that the compound's positioning and movement under
those conditions are altered with respect to the positioning and
movement that the compound would otherwise have.
[0117] In particular, delivery of a compound with respect to a
reference endpoint indicates the ability to control positioning and
movement of the compound so that the compound is eventually
positioned on the selected reference endpoint. In an in vitro
system, delivery of a compound is usually associated to a
corresponding modification of the chemical and/or biological
detectable properties and activities of the compound. In an in vivo
system, delivery of a compound is also typically associated with
modification of the pharmacokinetics and possibly pharmacodynamics
of the compound.
[0118] Pharmacokinetic of a compound indicates absorption,
distribution, metabolism and excretion of the compound from the
system, typically provided by the body of an individual. In
particular the term "absorption" indicates the process of a
substance entering the body, the term "distribution" indicates the
dispersion or dissemination of substances throughout the fluids and
tissues of the body, the term "metabolism" indicates the
irreversible transformation of parent compounds into daughter
metabolites and the term "excretion" indicates the elimination of
the substances from the body. If the compound is in a formulation,
pharmacokinetics also comprises liberation of the compound from the
formulation which indicates process of release of the compound,
typically a drug, from the formulation. The term "pharmacodynamic"
indicates physiological effects of a compound on the body or on
microorganisms or parasites within or on the body and the
mechanisms of drug action and the relationship between drug
concentration and effect. A skilled person will be able to identify
the techniques and procedures suitable to detect pharmacokinetics
and pharmacodynamic features and properties of a compound of
interest and in particular of an agent of interest such as a
drug.
[0119] The described method may be used to deliver therapeutic
agents to the brain of a subject, by loading the described
nanoparticles with a therapeutic agent of interest prior to
administration of the nanoparticle to the subject. Following
delivery of the loaded nanoparticle, the targeting agent will
facilitate delivery to a target cell of interest, such as a brain
endothelial cell. Following internalization by the target cell, the
nanoparticle will dissociate from the targeting agent. In the case
of brain endothelial cells, the internalized nanoparticle will then
be excreted from the cell into the interstitial space of the brain
where the particle will destabilize and secrete the loaded agent,
thereby delivering the agent to the brain or other target location.
In some embodiments the described methods may be carried out to
deliver a neurotransmitter such as serotonin or dopamine to the
brain, which may be used to treat a neurological disorder. Other
agents for use in treating neurological disorders may also be
delivered to the brain via the described methods. Imaging agents
that might not readily access the brain on their own may also be
delivered using the described methods. In some embodiments the
described methods may be used to deliver a nanoparticle carrying
the imaging agent Cu-64 to the brain of a subject to allow for
imaging. Further, the described methods may be used to deliver a
combination of one or more therapeutic agents, imaging agents, or
both therapeutic agents and imaging agent to the brain of a
subject.
[0120] The subject may be any animal, and preferably is a mammal
such as a mouse, rat, hamster, guinea pig, rabbit, cat, dog,
monkey, donkey, cow, horse, pig, and the like. Most preferably, the
mammal is a human. Importantly, the subject is one known to have a
condition that would benefit by the treatment being administered;
i.e., would benefit from delivery of the therapeutic agent to the
brain of the patient by systemic means (direct injection of the
therapeutic into the brain is one option, but the nanoparticle
delivery systems are intended to allow for the systemic delivery of
the therapeutics, and it is in this context that delivery is
envisioned). That is, such a patient has a disease or condition of
the brain where conventional treatments (i.e., without the use of
the nanoparticles described herein) are unable to deliver
sufficient quantities of the therapeutic agent systemically. This
pre-recognition of the disease or condition in the subject is
implicit in the definition of the subject or patient.
[0121] In some embodiments, subjects may be administered at least
one of the described nanoparticles in a daily dose range of 0.01
.mu.g to 500 mg per kg of the weight of the subject. The dose
administered to the subject may also be measured in terms of total
amount of at least one of the described nanoparticles administered
per day. In some embodiments, a subject is administered 5 to 5000
milligrams of at least one of the described nanoparticles per dose.
In some embodiments, a subject is administered up to 10 milligrams
of at least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 100 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 250 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 500 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 750 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 1000 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 1500 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 2000 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 2500 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 3000 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 3500 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 4000 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 4500 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, a subject is administered up to 5000 milligrams of at
least one of the described nanoparticles per dose. In some
embodiments, the described methods may be carried out so the
nanoparticles described herein is administered to a subject weekly,
bi-weekly, monthly, bi-month, semi-annually, or annually. Treatment
may be initiated with smaller dosages that are less than the
optimum dose followed by an increase in dosage over the course of
the treatment until the optimum effect under the circumstances is
reached.
[0122] When discussing the beneficial aspects of the ability of the
nanoparticles to deliver the therapeutic (or any other) agent
across the blood brain barrier (BBB), several criteria may be used.
Given the otherwise impenetrability of the BBB to therapeutics or
other agents, one such criterion is the ability of the
nanoparticle(s) to deliver the pharmaceutical agent in an amount to
be therapeutically useful, under conditions where the delivery of
the pharmaceutical agent by itself is unsuccessful in doing so. In
such embodiments, the methods may comprise the systemic
administration of a plurality of nanoparticles to a subject having
the neurological brain disorder (e.g., a disease such as cancer)
and in need of delivery of the therapeutic agent across a
blood-brain barrier to the subject's brain parenchyma, the
plurality of nanoparticles being administered at a dose rate
sufficient to enhance the delivery of the therapeutic agent to the
brain parenchyma in therapeutically useful quantities that are
higher than available using the free therapeutic agent alone. In
separate embodiments, depending on the efficiency of the brain
parenchyma in blocking the passage of the therapeutic agent alone,
this in increase in delivery of the therapeutic agent using the
nanoparticles can be at least 2 times, at least 5 time, at least 10
times, at least 100 times, or at least 1000 times higher than with
the therapeutic agent alone. In some cases, the therapeutic agent
may, by itself, be unable to cross the blood-brain barrier (or such
delivery is imperceptively small), such that any improvement in the
delivery of y such agent by means of the nanoparticle(s) disclosed
herein is considered to constitute delivery.
[0123] The nanoparticles described herein may be administered
orally in any acceptable dosage form such as capsules, tablets,
aqueous suspensions, solutions or the like. The nanoparticles may
also be administered parenterally including but not limited to:
subcutaneous, intravenous, intramuscular, intra-articular,
intra-synovial, and intrasternal injection or infusion techniques.
Alternatively, the nanoparticles will be administered intravenously
or intraperitoneally, for example, by injection. In independent
exemplary embodiments, the nanoparticles are administered by
intravenous and intracardiac injection.
[0124] The term "diluent" as used herein indicates a diluting agent
which is issued to dilute or carry an active ingredient of a
composition. Suitable diluents include any substance that can
decrease the viscosity of a medicinal preparation.
[0125] The term "excipient" as used herein indicates an inactive
substance used as a carrier for the active ingredients of a
medication. Suitable excipients for the pharmaceutical compositions
herein disclosed include any substance that enhances the ability of
the body of an individual to absorb the nanoparticle. Suitable
excipients also include any substance that can be used to bulk up
formulations with nanoparticles to allow for convenient and
accurate dosage. In addition to their use in the single-dosage
quantity, excipients can be used in the manufacturing process to
aid in the handling of nanoparticles. Depending on the route of
administration, and form of medication, different excipients may be
used. Exemplary excipients include but are not limited to
antiadherents, binders, coatings disintegrants, fillers, flavors
(such as sweeteners) and colors, glidants, lubricants,
preservatives, sorbents.
[0126] The term "functional group" as used herein indicates
specific groups of atoms within a molecular structure or portion
thereof that are responsible for the characteristic chemical
reactions of that structure or portion thereof. Exemplary
functional groups include hydrocarbons, groups containing halogen,
groups containing oxygen, groups containing nitrogen and groups
containing phosphorus and sulfur all identifiable by a skilled
person. Functional groups in the sense of the present disclosure
include a carboxylic acid, amine, triarylphosphine, azide,
acetylene, sulfonyl azide, thio acid and aldehyde. For example, a
functional group able to bind a corresponding functional group in a
targeting ligand can be selected to comprise the following binding
partners: carboxylic acid group and amine group, azide and
acetylene groups, azide and triarylphosphine group, sulfonyl azide
and thio acid, and aldehyde and primary amine. Additional
functional groups can be identified by a skilled person upon
reading of the present disclosure. As used herein, the term
"corresponding functional group" refers to a functional group that
can react to another functional group. Thus, functional groups that
can react with each other can be referred to as corresponding
functional groups.
[0127] An end-group indicates a constitutional unit that is an
extremity of a macromolecule or oligomer molecule. For example the
end-group of a PET polyester may be an alcohol group or a
carboxylic acid group. End groups can be used to determine molar
mass. Exemplary end groups comprise --OH, --COOH, NH.sub.2, and
OCH.sub.3.
[0128] The term "leaving group," as understood by persons skilled
in the art of organic synthesis, refers to a functional group
susceptible to displacement by a nucleophile. Leaving groups can be
anions or neutral moieties, but in either case it is crucial that
the leaving group be able to stabilize the additional electron
density that results from bond heterolysis. Exemplary leaving
groups include halides such as chloride, bromide, and iodide,
sulfonate esters such as mesylate and tosylate, water, and cationic
amines.
[0129] The term "ligand" or "targeting ligand" as used in the
present disclosure indicates any molecule that can be presented on
the surface of a nanoparticle for the purpose of engaging a
specific target, and in particular specific cellular recognition,
for example by enabling cell receptor attachment of the
nanoparticle. Examples of suitable ligands include, but are not
limited to, vitamins (e.g. folic acid), proteins (e.g. transferrin,
monoclonal antibodies), monosaccharides (e.g. galactose), peptides,
and polysaccharides. In particular targeting ligands can be
antibodies against certain surface cell receptors such as
transferrin receptor ("TfR").
[0130] The term "nanoparticle" as used herein indicates a composite
structure of nanoscale dimensions. In particular, nanoparticles are
typically particles of a size in the range of from about 1 to about
1000 nm and are usually spherical although different morphologies
are possible depending on the nanoparticle composition. The portion
of the nanoparticle contacting an environment external to the
nanoparticle is generally identified as the surface of the
nanoparticle. In nanoparticles herein described, the size
limitation can be restricted to two dimensions and so that
nanoparticles herein described include composite structure having a
diameter from about 1 to about 1000 nm, where the specific diameter
depends on the nanoparticle composition and on the intended use of
the nanoparticle according to the experimental design. For example,
nanoparticles to be used in several therapeutic applications
typically have a size of about 200 nm or below. In certain
embodiments where the nanoparticles are for delivery associated to
cancer treatment typically have a diameter from about 80 nm to
about 120 nm.
[0131] Additional desirable properties of the nanoparticle(s), such
as surface charges and steric stabilization, can also vary in view
of the specific application of interest. Properties of the
particles may be understood by a skilled person upon reading of the
present disclosure. Nanoparticle dimensions and properties can be
detected by techniques known in the art. Exemplary techniques to
detect particles dimensions include but are not limited to dynamic
light scattering (DLS) and nanoparticle tracking analysis (NTA) and
a variety of microscopies such at transmission electron microscopy
(TEM) and atomic force microscopy (AFM). Exemplary techniques to
detect particle morphology include but are not limited to TEM and
AFM. Exemplary techniques to detect surface charges of the
nanoparticle include but are not limited to zeta potential method.
Additional techniques suitable to detect other chemical properties
comprise by .sup.1H, .sup.11B, .sup.13C and .sup.19F NMR, UV/Vis
and infrared/Raman spectroscopies and fluorescence spectroscopy
(when nanoparticle is used in combination with fluorescent labels)
and additional techniques identifiable by a skilled person
[0132] "Optional" or "optionally" means that the subsequently
described circumstance may or may not occur, so that the
description includes those embodiments where the circumstance
occurs and instances where it does not. For example, in the phrase
"at least one small molecule therapeutic agent, optionally linked
to the nanoparticle core by way of an optional linker," the term
"optionally linked" is interpreted to mean that the therapeutic
agent may or may not be linked to the nanoparticle, and embraces
both embodiments in the alternative. Similarly, the term "optional
linker" embraces both embodiments, in the alternative, where the
linker is either present or absent (in this case, since the
optional linker refers to the linking of the therapeutic agent and
the nanoparticle core, where the linker is absent, the inkage is a
direct bond.
[0133] The term a "polymer" as used at least in the context of the
nanoparticle core, herein embraces its generally recognized meaning
and indicates a large molecule composed of repeating structural
units typically connected by covalent chemical bonds. A suitable
polymer may be a linear and/or branched and can take the form of a
homopolymer or a co-polymer. If a co-polymer is used, the
co-polymer may be a random copolymer or a branched co-polymer.
Exemplary polymers comprise water-dispersible and, in particular,
water-soluble polymers. For example, suitable polymers include, but
are not limited to polysaccharides, polyesters, polyamides,
polyethers, polycarbonates, polyacrylates, etc. For therapeutic
and/or pharmaceutical uses and applications, the polymer should
have a low toxicity profile and that are not toxic or cytotoxic.
Suitable polymers include polymers having a molecular weight of
about 500,000 or below. Suitable polymers can have a molecular
weight of about 100,000 and below.
[0134] The term "polymer containing a boronic acid" or a "linker
having a boronic acid" and the like as used herein indicate
containing at least one boronic acid group presented for binding to
a hydroxyl group of a polymer containing polyols. In particular,
polymers containing boronic acids of the nanoparticles herein
described include a polymer comprising in at least one structural
unit an alkyl or aryl substituted boronic acid containing a carbon
to boron chemical bond. Suitable boronic acid polymers comprise
polymers wherein boronic acid is in a terminal structural unit or
in any other suitable position to provide the resulting polymer
with hydrophilic properties. In this regard, the (nitro)phenyl
boronic acids, and especially nitrophenyl boronic acids, are
preferred. As used herein, in the context of "(nitro)phenylboronic
acid," the parenthetical "nitro" reflects separate embodiments
where the phenylboronic acid moiety has at least one nitro group
appended to the phenyl group (i.e., a nitrophenylboronic acid
moiety) and where the phenylboronic acid group does not contain a
nitro group (i.e., a phenylboronic acid moiety).
[0135] The term "polymer containing a polyol" or "polyol-containing
polymer" refers to a polymer refers to a polymer presenting
multiple hydroxyl functional groups. These multiple hydroxyl groups
can serve to improve the hydrophilicity of the core, making the it
better dispersible in aqueous media and/or may provide sites for
linking the various linking groups described herein. In certain
embodiments, the polymer containing a polyol suitable to form the
nanoparticles here described comprise polymers presenting at least
a portion of the hydroxyl functional groups for a coupling
interaction with at least one boronic acid of a polymer containing
a boronic acid.
[0136] In certain embodiments, the polyols comprise monomeric
polyols such as pentaerythritol, ethylene glycol, glycerin, and
various sugars, including mucic acid. Exemplary polymers containing
polyols comprise polyesters, polyethers and polysaccharides.
Exemplary suitable polyethers include but are not limited to diols,
such as polyethylene glycol, polypropylene glycol, and
poly(tetramethylene ether) glycol. Exemplary, suitable
polysaccharides include but are not limited to cyclodextrins,
starch, glycogen, cellulose, chitin and .beta.-Glucans. Exemplary,
suitable polyesters include but are not limited to polycarbonate,
polybutyrate and polyethylene terephthalate, all terminated with
hydroxyl end groups. Exemplary polymers containing polyols comprise
polymers of about 500,000 or less molecular weight and in
particular from about 300 to about 100,000.
[0137] The term "polypeptide" as used herein indicates an organic
linear, circular, or branched polymer composed of two or more amino
acid monomers and/or analogs thereof. The term "polypeptide"
includes amino acid polymers of any length including full length
proteins and peptides, as well as analogs and fragments thereof. A
polypeptide of three or more amino acids is also called a protein
oligomer, peptide or oligopeptide. In particular, the terms
"peptide" and "oligopeptide" usually indicate a polypeptide with
less than 50 amino acid monomers. As used herein the term "amino
acid", "amino acidic monomer", or "amino acid residue" refers to
any of the twenty naturally occurring amino acids, non-natural
amino acids, and artificial amino acids and includes both D an L
optical isomers. In particular, non-natural amino acids include
D-stereoisomers of naturally occurring amino acids (these including
useful ligand building blocks because they are not susceptible to
enzymatic degradation). The term "artificial amino acids" indicate
molecules that can be readily coupled together using standard amino
acid coupling chemistry, but with molecular structures that do not
resemble the naturally occurring amino acids. The term "amino acid
analog" refers to an amino acid in which one or more individual
atoms have been replaced, either with a different atom, isotope, or
with a different functional group but is otherwise identical to
original amino acid from which the analog is derived. All of these
amino acids can be synthetically incorporated into a peptide or
polypeptide using standard amino acid coupling chemistries. The
term "polypeptide" as used herein includes polymers comprising one
or more monomer or building blocks other than an amino acid
monomer. The terms monomer, subunit, or building blocks indicate
chemical compounds that under appropriate conditions can become
chemically bonded to another monomer of the same or different
chemical nature to form a polymer. The term "polypeptide" is
further intended to comprise a polymer wherein one or more of the
building blocks is covalently bound to another by a chemical bond
other than amide or peptide bond.
[0138] The term "protein" as used herein indicates a polypeptide
with a particular secondary and tertiary structure that can
participate in, but not limited to, interactions with other
biomolecules including other proteins, DNA, RNA, lipids,
metabolites, hormones, chemokines, and small molecules Exemplary
proteins herein described are antibodies. The terra protein
embraces fusion proteins.
[0139] The term "prevent" as used herein to describe the action of
inhibiting cell proliferation or the growth of tumors, or
ameliorating the symptoms, prolonging the survival of, or otherwise
mitigating the undesirable effects of the disease for which the
patient is being treated.
[0140] The distinction between the terms "small molecule
therapeutic" and "large molecule therapeutic" should be apparent to
a person of skill in the art, as based on molecular weight of the
molecule. In certain embodiments, the demarcation may be defined as
a molecular weight of about 600 Da, 800 Da, 1000 Da, or 1200 Da. In
the context of "small molecules capable of passing the blood brain
barrier," unless the molecule is specifically known to be incapable
of passing the blood brain barrier, this is normally defined by a
"Rule of Five," typically attributed to Lipinski, that describes
the physical parameters seen as necessary or essential for a small
molecule to pass. According to this Rule, a good absorption and
permeability is likely if:
[0141] Molecular weight is .ltoreq.600;
[0142] Oil/water distribution coefficient (Log P) is .ltoreq.5;
[0143] Hydrogen bond donors .ltoreq.3-5 (expressed as the sum of
OHs and NHs);
[0144] Hydrogen bond acceptor .ltoreq.7-10 (expressed as the sum of
Ns and Os); and
[0145] Number of rotatable bonds .ltoreq.5-10.
For the purposes of this disclosure, a small molecule therapeutic
will be considered capable of passing the blood brain barrier
without the disclosed nanoparticle systems if it is otherwise known
to pass or meets these criteria. Likewise, unless otherwise stated,
a small molecule will be deemed incapable of passing through the
blood brain barrier to deliver therapeutically effect amounts
without the nanoparticles set forth herein, if it does not satisfy
these criteria. Against this context, a review by H. Pajouhesh, et
al., "Medicinal Chemical Properties of Successful Central Nervous
System Drugs" NeuroRx, 2005 October; 2(4) 541-553, which is
incorporated by reference herein for its teaching in this regard,
based on literature surveys concluded that successful CNS drugs
had:
[0146] Molecular weight--preferably below a 400 to 600-Da
range;
[0147] Oil/water distribution coefficient (Log P)--preferably in
the range of 1.5-2.7;
[0148] Hydrogen bond donors and acceptors--collectively
(O+N)=4.32:2.12 hydrogen bond acceptors and 1.5 hydrogen bond
donors;
[0149] PSA (polar surface area) (defined as the surface area
(A.sup.2) occupied by nitrogen and oxygen atoms and the polar
hydrogens attached to them and is strongly reflective of hydrogen
bonding capacity and polarity--less than 60-70 .ANG..sup.2
[0150] Aqueous solubility >60 .mu.g/ml;
[0151] Number of rotatable bonds--less than 5.
The number of (OH+NH) bonds (.ltoreq.1.5 to 2), number of (O+N)
bonds (.ltoreq.4.3 to 6), hydrogen bond acceptors (.ltoreq.2.5 to
4), and rotatable bonds (.ltoreq.4.7 to 7) appeared to distinguish
CNS (74) and gastrointestinal-metabolic (38) drugs marketed between
1983 and 2002, suggesting these parameters are most important.
Generally, marketed CNS drugs tend to smaller with a more compact
and less flexible structure, the surface having fewer polar groups
able to function as hydrogen bond donors and acceptors and compared
to the total surface area, the PSA is reduced. These can give
further definition to the small molecules capable of passing the
blood brain barrier.
[0152] The term "substantially free" as in "where the nanoparticle
core and/or surface is substantially free of cationic mucic-acid
containing polymer (cMAP)" refers to a composition in which the
core or surface composition contains less than 3 mol % of the cMAP
polymer, preferably less than 1%, and more preferably no cMAP
polymer. Unless otherwise indicated, the term refers to the absence
of cMAP polymer.
[0153] The term "target" as used herein indicates a biological
system of interest including organs, tissues, or any portion
thereof and may include in vitro or in vivo biological systems or
any portion thereof. In certain embodiments, the target is a tumor
or cancer cells within the brain itself.
[0154] As used herein, the terms "therapeutic agent" and
"chemotherapeutic agent" are intended to mean a compound that, when
present in a therapeutically effective amount, produces a desired
therapeutic effect on a patient; for example, utilized to treat,
combat, ameliorate, prevent or improve an unwanted condition or
disease of a patient or subject. In the case of cancer, this
includes stopping the progression of the cancer and/or a reduction
in the presence of corresponding tumors or cancer cells. Such
inhibition may occur for example, and without limitation, via a
direct interaction, and/or through a competitive interaction, or
via an allosteric interaction with a corresponding receptor. The
term "[chemo]therapeutic agent" is intended to embrace the separate
embodiments of the general term "therapeutic agent" and the more
specific "chemotherapeutic agent," the latter term generally
associated with cytotoxic agents for use in chemotherapy for
cancer. More generally, the use of brackets or parentheses, in this
way, is intended to refer to separate embodiments where the
bracketed or parenthetical text is both present and absent.
[0155] In a addition to the therapeutic agents disclosed elsewhere
herein, the term more generally encompasses any lipophilic or
hydrophilic, synthetic or naturally occurring biologically active
therapeutic agent including those known in the art. The Merck
Index, An Encyclopedia of Chemicals, Drugs, and Biologicals, 13th
Edition, 2001, Merck and Co., Inc., Whitehouse Station, N.J.
Examples of such therapeutic agents include, but are not limited
to, small molecule pharmaceuticals, antibiotics, steroids,
polynucleotides (e.g. genomic DNA, cDNA, mRNA, siRNA, shRNA, miRNA,
antisense oligonucleotides, viruses, and chimeric polynucleotides),
plasmids, peptides, peptide fragments, small molecules (e.g.
doxorubicin), chelating agents (e.g. deferoxamine,
ethylenediaminetetraacetic acid (EDTA)), natural products (e.g.
Taxol, Amphotericin), and other biologically active macromolecules
such as, for example, proteins and enzymes. See also U.S. Pat. No.
6,048,736 which lists active agents (therapeutic agents) that can
be used as therapeutic agent with nanoparticles herein described
Small molecule therapeutic agents may not only be the therapeutic
agent within the composite particle but, in an additional
embodiment, may be covalently bound to a polymer in the composite.
In several embodiments, the covalent bond is reversible (e.g.
through a prodrug form or biodegradable linkage such as a
disulfide) and provides another way of delivering the therapeutic
agent. In several embodiments therapeutic agent that can be
delivered with the nanoparticles herein described include
chemotherapeutics such as epothilones, camptothecin-based drugs,
taxol, or a nucleic acid such as a plasmid, siRNA, shRNA mRNA,
antisense oligonucleotides aptamers or their combination, and
additional drugs identifiable by a skilled person upon reading of
the present disclosure
[0156] A "therapeutically effective amount" or "effective amount"
of a composition is a predetermined amount calculated to achieve
the desired effect. The activity contemplated by the present
methods includes both medical therapeutic and/or prophylactic
treatment, as appropriate. As used herein, "therapeutically
effective amount" refers to the amount of active compound or
pharmaceutical agent that elicits a biological or medicinal
response in a tissue, system, animal, individual or human that is
being sought by a researcher, veterinarian, medical doctor or other
clinician, which includes one or more of the following as specified
in the particular methodology: (1) preventing the disease; for
example, preventing a disease, condition or disorder in an
individual that may be predisposed to the disease, condition or
disorder but does not yet experience or display the pathology or
symptomatology of the disease, (2) inhibiting the disease; for
example, inhibiting a disease, condition or disorder in an
individual that is experiencing or displaying the pathology or
symptomatology of the disease, condition or disorder (i.e.,
arresting further development of the pathology and/or
symptomatology), and (3) ameliorating the disease; for example,
ameliorating a disease, condition or disorder in an individual that
is experiencing or displaying the pathology or symptomatology of
the disease, condition or disorder (i.e., reducing the severity of
the pathology and/or symptomatology).
[0157] As used herein, the term "transferrin" (abbreviated "Tf") is
meant to encompass variants and isoforms of the protein, as well as
fragments of the protein capable of binding to the transferrin
receptor ("TfR"). For example, the term would include
holo-transferrin as well as transferrin itself.
[0158] The terms "treat," "treated," or "treating" as used herein
refers to both therapeutic treatment and prophylactic or
preventative measures, wherein the object is to prevent or slow
down (lessen) an undesired physiological condition, disorder or
disease, or to obtain beneficial or desired clinical results. For
the purposes of this invention, beneficial or desired clinical
results include, but are not limited to, diminishment of the extent
of the condition, disorder or disease; stabilization (i.e., not
worsening) of the state of the condition, disorder or disease;
delay in onset or slowing of the progression of the condition,
disorder or disease; amelioration of the condition, disorder or
disease state; and remission (whether partial or total), whether
detectable or undetectable, or enhancement or improvement of the
condition, disorder or disease. Treatment includes eliciting a
clinically significant response with or without excessive levels of
side effects.
[0159] The term "vehicle" as used herein indicates any of various
media acting usually as solvents, carriers, binders, excipients or
diluents for a nanoparticle comprised in the composition as an
active ingredient.
GENERAL EMBODIMENTS OF THE PRESENT DISCLOSURE
[0160] The following listing of embodiments is intended to
complement, rather than displace or supersede, the previous
descriptions.
Embodiment 1
[0161] A polymer, polymer conjugate, or nanoparticle composition
comprising:
[0162] a polymer or nanoparticle core to which is bonded
[0163] (a) at least one targeting agent, the targeting agent
comprising a ligand attached to an external surface of the
nanoparticle core by a linker; [0164] the ligand having an affinity
for binding to a receptor expressed by endothelial cells of the
blood-brain barrier; and [0165] the linker being cleavable when
subject to conditions inside an endothelial cell of the blood-brain
barrier, wherein the cleavage comprises hydrolysis, chemical
reduction, or enzymatic cleavage of the linker; and one or both
of
[0166] (b) at least one small molecule therapeutic agent,
optionally linked to the nanoparticle core by way of an optional
linker; and/or
[0167] (c) at least one large molecule therapeutic agent, linked to
the nanoparticle by way of an optional linker;
[0168] wherein the nanoparticle comprises polyol-containing
polymer, such as a mucic acid-containing polymer (MAP) or a
cationic mucic acid polymer (cMAP), a peptide, a
poly(lactic-co-glycolic acid) (PLGA) polymer, chitosan, a synthetic
polymer such as a polyamide, a polycarbonate, a polyethylene, a
polyethyleneimine, gold, and/or iron oxide, or any combination
thereof, and further wherein
[0169] the large molecule therapeutic agent, when present, and the
targeting agent comprise different chemical entities. Additionally,
or alternatively, in certain independent Aspects of this
Embodiment, the presence of a cationic mucic acid polymer (cMAP) is
specifically excluded.
[0170] Additionally, or alternatively, in certain independent
Aspects of this Embodiment, the composition is or can be
independently characterized as a polymer (i.e., without regard to
any specific second order structure) comprising the single polymer
or more polymers, for example a mucic acid polymer linked to a
small molecule therapeutic agent.
[0171] Additionally, or alternatively, in certain independent
Aspects of this Embodiment, the composition is or can be
characterized as a polymer chain (i.e., without regard to any
specific second order structure) comprising the coupling of two or
more polymers, for example a mucic acid polymer and a
(nitro-boronic acid polymer).
[0172] Additionally, or alternatively, in certain independent
Aspects of this Embodiment, the composition is or can be
characterized as a nanoparticle or population of nanoparticles.
[0173] Additionally, or alternatively, in certain independent
Aspects of this Embodiment, the core is independently characterized
simply as a polymer. In other independent Aspects of this
Embodiment, the core is independently characterized as a
nanoparticle.
[0174] Additionally, or alternatively, in certain independent
Aspects of this Embodiment, the polymer, polymer conjugate, or
nanoparticle composition comprises the polymer or nanoparticle core
to which is bonded at least one targeting agent and the at least
one small molecule therapeutic agent.
[0175] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the polymer, polymer conjugate, or nanoparticle
composition comprises the polymer or nanoparticle core to which is
bonded at least one targeting agent and the at least one large
molecule therapeutic agent.
[0176] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the polymer, polymer conjugate, or nanoparticle
composition comprises the polymer or nanoparticle core to which is
bonded at least one targeting agent, the at least one small
molecule therapeutic agent, and the at least one large molecule
therapeutic agent.
[0177] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the linker comprises a polyethylene glycol
(PEG) polymer moiety conjugated to the surface of the nanoparticle
core by a pH sensitive linkage selected from the group consisting
of an acetal, a borate ester, a carbonate, a carboxylic acid ester,
a diamino ketal, a disulfide, a hydrazone, an imine, a ketal, an
orthoester, or a peptide linkage.
[0178] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the linker comprises, when the pH sensitive
linkage is carboxylic acid ester linkage, the surface of the
nanoparticle core comprises poly(lactic-co-glycolic acid) (PLGA)
polymers, wherein the linker is dissociable when subject to pH
conditions inside an endothelial cell of the blood-brain
barrier.
[0179] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the linker comprises a polyethylene glycol
(PEG) polymer moiety conjugated to the surface of the nanoparticle
core by a peptide linkage, wherein the peptide linkage can be
enzymatically cleaved to cause dissociation of the ligand from the
nanoparticle when the nanoparticle is inside a brain endothelial
cell.
[0180] Additionally, or alternatively, in other independent Aspects
of this Embodiment, one or both of the therapeutic agents
characterized as small molecule therapeutic agent and the large
molecule therapeutic agent is a small molecule chemotherapeutic
agent and large molecule chemotherapeutic agent, respectively.
[0181] Additionally, or alternatively, in other independent Aspects
of this Embodiment, the polymer, polymer conjugate, or nanoparticle
composition comprises a structure as described in any one of the
structures described in U.S. Pat. Nos. 8,557,292; 8,746,999;
8,968,714; 9,186,327; 9,334,367; 9,610,355; or 9,913,911
(CIT-5200); PCT application PCT/US2009/053620 (CIT-5200); U.S. Pat.
No. 9,468,681; PCT application PCT/US2013/028663 (CIT-6456); U.S.
Pat. No. 9,132,097; PCT application PCT/US2013/028681 (CIT-6455);
U.S. patent application Ser. No. 14/120,309; PCT application
PCT/US2014/000099 (CIT-6566); U.S. patent application Ser. No.
15/180,201; or PCT application PCT/US2016/037166 (CIT-7222). These
references are incorporatede by reference herein for all purposes,
but at least for the descriptions of the nanoparticles and uses
thereof described therein.
Embodiment 2
[0182] The polymer, polymer conjugate, or nanoparticle composition
of Embodiment 1, wherein the polymer or nanoparticle core
comprises, consists essentially of, or consists of mucic acid
polymer (MAP), a cationic mucic acid polymer (cMAP), a
poly(lactic-co-glycolic acid) (PLGA), chitosan, polyethyleneimine,
polysaccharide, polyester, polyamide, polyether, polycarbonate,
polyacrylate, iron oxide, or gold. Each of these materials
constitute independent Aspects of this Embodiment.
[0183] Additionally, or alternatively, other water-dispersible and
in particular water-soluble polymers may be considered for this
purpose. For therapeutic and/or pharmaceutical uses and
applications, the polymer should have a low toxicity profile and in
particular that are not toxic or cytotoxic. Additionally, or
alternatively, such polymers include those having a molecular
weight of about 500,000 or below. In particular, suitable polymers
can have a molecular weight of about 100,000 and below.
[0184] Further, in additional or alternative Aspects of this
Embodiment, the polymer or nanoparticle core by itself (i.e.,
without the targeting ligand, or small or large molecule
therapeutic compounds), as further characterized herein, is
considered an independent aspect of this Embodiment.
Embodiment 3
[0185] The polymer, polymer conjugate, or nanoparticle composition
of Embodiment 1 or 2, wherein the polymer or nanoparticle core
comprises, consists essentially of, or consists of a
polyol-containing polymer, preferably a sugar-containing polymer,
for example, a polymer derived from glucose, fructose, mannitol,
mucic acid, sucrose, galactose, sorbitol. xylose or galactose, more
preferably from mucic acid. Additionally, or alternatively, in
certain Aspects of this Embodiment, the polyol-containing polymer
of the polymer, polymer conjugate, or nanoparticle composition
further comprises dihydroxy, hydroxy-amine, and/or diamine linkages
between the polyol moieties, wherein the dihydroxy, hydroxy-amine,
and/or diamine linkages independently comprise 1 to 6 carbon atoms
between the respective hydroxy and amine functional groups.
Embodiment 4
[0186] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 3, wherein the polymer or
nanoparticle core is charge neutral and/or negatively charged. In
certain Aspects of this Embodiment, the polymer or nanoparticle
core contains no cationic moieties (e.g., does not contain any
cationic mucic acid polymers (cMAP) or polymer portions.
Embodiment 5
[0187] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 3, wherein the polymer or
nanoparticle core comprises at least one cationic and/or at least
one anionic moiety. Additionally, or alternatively, in independent
Aspects of this Embodiment, the polymer or nanoparticle core
contains a net cationic charge, a net anionic charge, or is
charge-balanced neutral. In certain independent Aspects within this
Embodiment, a net cationic charge may be provided by the
incorporation of amidines, quaternary ammoniums, primary,
secondary, or tertiary amine groups (protonated below their pKa's),
and immidazolium groups. In certain independent Aspects within this
Embodiment, a net anionic charge may be provided by the
incorporation of functional groups including sulfonates, nitrates,
carboxylates, and phosphonates.
Embodiment 6
[0188] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 5, wherein the polymer or
nanoparticle core independently comprises units of the formula:
##STR00016##
Embodiment 7
[0189] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 6, wherein the polymer or
nanoparticle core comprises a polymer containing a polyol, wherein
the polymer containing the polyol is derived from the coupling of a
compound of Formula A with a compound of Formula B; wherein the
compound of Formula A is:
##STR00017##
wherein n is a number from 1 to 20; and
[0190] R is a functional group residue (e.g., --COOH, --NH.sub.2,
--OH) corresponding to that of an amino acid containing a third
functional group, for example arginine (R is
CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2.sup.+), histidine (R is
CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof; and
[0191] the compound of Formula B is:
##STR00018##
in which
[0192] q is a number from 1 to 20;
[0193] p is a number from 20 to 200; and
[0194] L is a leaving group.
[0195] Additionally, or alternatively, in independent Aspects of
this Embodiment, the compound of Formula B is:
##STR00019##
Additionally, or alternatively, in some Aspects of this Embodiment,
the compound of Formula B specifically excludes:
##STR00020##
[0196] Additionally, or alternatively, in independent Aspects of
this Embodiment, the compound of Formula B is:
##STR00021##
Embodiment 8
[0197] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 7, wherein the polymer or
nanoparticle core independently comprises units of the formula:
##STR00022##
Embodiment 9
[0198] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 8, wherein the polymer or
nanoparticle core independently comprises units of the formula:
##STR00023##
[0199] Additionally, or alternatively, in independent Aspects of
any one of Embodiments 7 to 9, q is a number from 1 to 20 and p is
a number from 20 to 200. Additionally, or alternatively, in other
independent Aspects any one of Embodiments 7 to 9, n is 1 and q is
1.
Embodiment 10
[0200] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 9, wherein the polymer or
nanoparticle core independently comprises units derived from the
coupling of:
##STR00024##
where n is a number from 1 to 20 with a trifunctional amino acid,
for example including arginine, histidine, lysine, aspartic acid,
glutamic acid, serine, threonine, asparagine, or glutamine, or a
moiety derived therefrom, preferably from aspartic acid, or a
moiety derived therefrom.
Embodiment 11
[0201] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 10, wherein the polymer or
nanoparticle core polymer comprises a polyol and a polyethylene
oxide linkage:
##STR00025##
where x is in a range of from 15 to 100, preferably 20 to 85.
Embodiment 12
[0202] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 11, wherein the polymer or
nanoparticle core polymer comprises a polyol structure:
##STR00026##
where y is in a range of from 10 to 25, preferably 15 to 25, more
preferably about 20, or more broadly:
##STR00027##
where R is a functional group residue (containing, e.g., a --COOH,
--NH.sub.2, --OH residue) corresponding to that of an amino acid
containing a third functional group, for example arginine (R is
CH.sub.2CH.sub.2CH.sub.2NHC(NH.sub.2).sub.2+), histidine (R is
CH.sub.2-imidazolyl), lysine (R is
CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--NH.sub.2), aspartic acid (R
is CH.sub.2--COOH), glutamic acid (R is CH.sub.2--CH.sub.2--COOH),
serine (R is CH.sub.2--OH), threonine (R is CH(OH)(CH.sub.3))
asparagine (R is CH.sub.2--C(O)NH.sub.2), glutamine (R is
CH.sub.2--CH.sub.2--C(O)NH.sub.2), tyrosine (R is CH.sub.2-Ph-OH),
tryptophan (R is CH.sub.2-indolyl), or a salt thereof.
[0203] Additionally, or alternatively, in certain Aspects of this
Embodiment, R is independently linked to one or more of the
targeting agent, the small molecule [chemo]therapeutic agent,
and/or the large molecule [chemo]therapeutic agent.
Embodiment 13
[0204] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 12, wherein the polymer or
nanoparticle core copolymer comprises residues of lactic acid,
glycolic acid, or a combination thereof. In certain additional or
alternative Aspects of this Embodiment, the residues of the lactic
acid, glycolic acid, or a combination thereof are arranged as
polymers or co-polymers of these materials. Additionally, or
alternatively, the polymer or nanoparticle core copolymer consists
of these polymers or co-polymers. Additionally, or alternatively,
the polymer or nanoparticle core copolymer comprise one or more
sugar polyol (including mucic acid), lactic acid, and/or glycolic
acid residues. That is, in some Aspects of this Embodiment,
poly(lactic-co-glycolic acid) (PLGA) core is further modified to
incorporate sugars having suitable hydroxyl groups to allow the
particle to be conjugated to a boronic acid-containing targeting
agent.
Embodiment 14
[0205] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 13, wherein the polymer or
nanoparticle core polymer comprises alternating charged and
uncharged segments comprising one or more of the following
structural units of Formula (I) or Formula (II) or Formula
(III):
##STR00028##
wherein
[0206] A is an uncharged segment comprising polyalkylene
glycol;
[0207] B is a cationically charged segment comprising at least one
polyhydroxy linkage comprising at least one pair of vicinal diols.
In certain subset embodiments, A and B independently have number
average molecular weights in a range 500 Da to about 5000 Da,
greater than 5000 da to about 10 kDa, greater than 10 kDa to about
20 kDa, greater than 20 kDa to about 30 kDa, greater than 30 kDa to
about 40 kDa, greater than greater than 40 kDa to about 50 kDa, or
any combination thereof. In other subsets, either A or B, or both A
and B have a number average molecular weight in a range of greater
than 5000 Da to about 50,000 Da.
Embodiment 15
[0208] The polymer, polymer conjugate, or nanoparticle composition
of Embodiment 14, wherein A is or comprises polyethylene glycol and
an appropriate linking group.
Embodiment 16
[0209] The polymer, polymer conjugate, or nanoparticle composition
of Embodiment 15 or 15, wherein the polyalkylene glycol has a
nominal number averaged molecular weight in a range of from about
500 Daltons to about 50,000 Daltons. Additionally, or
alternatively, in certain Aspects of this Embodiment, the
polyalkylene glycol has a nominal number averaged molecular weight
in a range of from about 500 Da to about 1 kDa, greater than 1 kDa
to about 5 kDa, greater than 5 kDa to about 10 kDa, greater than 10
kDa to about 15 kDa, greater than 15 kDa to about 20 kDa, greater
than 20 kDa to about 30 kDa, greater than 30 kDa to about 40 kDa,
greater than 40 kDa to about 50 kDa, or any combination of two or
more of these ranges.
Embodiment 17
[0210] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 16, where B is a cationically
charged segment comprising at least one polyhydroxy sugar linkage
comprising at least one pair of vicinal diols.
Embodiment 18
[0211] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 17, wherein B comprises at least
one repeating subunit comprising a structure:
##STR00029##
Note that the structure designated as
##STR00030##
is functionally equivalent to that presented immediately preceding
and this representation is intended to refer to both.
Embodiment 19
[0212] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 18, wherein B further comprises at
least one repeating subunit comprising a structure:
##STR00031##
wherein m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, preferably 4-6 or
5.
Embodiment 20
[0213] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 19, wherein B comprises at least
one repeating subunit comprising cMAP, whose subunit structure is
represented as:
##STR00032##
wherein
[0214] m is independently at each occurrence 1, 2, 3, 4, 5, 6, 7,
8, 9, or 10, preferably 4-6 or 5; and
[0215] n is independently at each occurrence 1, 2, 3, 4, or 5. In
other related embodiments, m and n can be larger, for example to
about 10.
Embodiment 21
[0216] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 20, described by a structure:
##STR00033##
wherein
[0217] Chain A is
##STR00034##
[0218] Chain B is
##STR00035##
[0219] cMAP is
##STR00036##
[0220] p and q are sufficient to provide a number average molecular
weight for the subunits comprising cMAP and PEG, respectively,
independently in a range of from about 500 Da to about 50,000
Da;
[0221] m is independently at each occurrence 1, 2, 3, 4, 5, 6, 7,
8, 9, or 10, preferably 4-6 or 5;
[0222] n and r are independently at each occurrence 0, 1, 2, 3, 4,
or 5; and
[0223] X.sub.1 and X.sub.2 are independently at each occurrence
C.sub.1-6 alkyl, optionally substituted with --OH, --COOH,
--C(.dbd.O)O(alkyl), --C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl),
--N(alkyl).sub.2, or a salt or protected analog thereof.
[0224] Note that, to meet the molecular weight restrictions, the
numeric values for p corresponds to a range of from about 1 to
about 100, preferably from about 10 to about 100, and the numeric
values for q corresponds to as range from about 12 to about 1200.
In subsets of these embodiments, q can also be in a range of from
about 100 to about 500. In certain subsets of this Embodiment, p
and q are sufficient to provide a number average molecular weight
for the subunits comprising cMAP and PEG, respectively,
independently in a range of from about 500 Da to about 1000 Da,
greater than 1000 Da to about 5000 Da, greater than 5000 Da to
about 10,000 Da, greater than 10,000 to about 25,000 Da, greater
than 25,000 Da to about 50,000 Da, or any combination of two or
more of these ranges, as well as the corresponding numeric values
for these MW.sub.n ranges. In other subsets of this Embodiment,
X.sub.1 and X.sub.2 are independently --(CH.sub.2).sub.1-4--COOH
and --(CH.sub.2).sub.1-4--NH.sub.2.
Embodiment 22
[0225] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 21, described by a structure:
##STR00037##
wherein
[0226] cMAP is
##STR00038##
[0227] Chain B is
##STR00039##
[0228] Chain C is
##STR00040##
[0229] End Group D is:
##STR00041##
[0230] p and q are sufficient to provide a number average molecular
weight for the subunits comprising cMAP and PEG, respectively,
independently in a range of from about 500 Da to about 50,000
Da;
[0231] m is independently at each occurrence 1, 2, 3, 4, 5, 6, 7,
8, 9, or 10, preferably 4-6 or 5;
[0232] n and r are independently at each occurrence 0, 1, 2, 3, 4,
or 5;
[0233] z is equal to or greater than 1 (up to about 2, 4, 6, 8, or
10); and
[0234] X.sub.2 is independently at each occurrence C.sub.1-6 alkyl,
optionally substituted with --OH, --COOH, --C(.dbd.O)O(alkyl),
--C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl), --N(alkyl).sub.2, or a
salt or protected analog thereof, and.
[0235] X.sub.3 is --OH, --COOH, --C(.dbd.O)O(alkyl),
--C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl), --N(alkyl).sub.2, or a
salt or protected analog thereof.
[0236] Again, as in Embodiment 21, to meet the molecular weight
restrictions, the numeric values for p corresponds to a range of
from about 1 to about 100, preferably from about 10 to about 100,
and the numeric values for q corresponds to as range from about 12
to about 1200. In subsets of these embodiments, q can also be in a
range of from about 100 to about 500. In certain subsets of this
Embodiment, p and q are sufficient to provide a number average
molecular weight for the subunits comprising cMAP and PEG,
respectively, independently in a range of from about 500 Da to
about 1000 Da, greater than 1000 Da to about 5000 Da, greater than
5000 Da to about 10,000 Da, greater than 10,000 to about 25,000 Da,
greater than 25,000 Da to about 50,000 Da, or any combination of
two or more of these ranges, as well as the corresponding numeric
values for these MW.sub.n ranges. In other subsets of this
Embodiment, X.sub.1 and X.sub.2 are independently
--(CH.sub.2).sub.1-4--COOH and --(CH.sub.2).sub.1-4--NH.sub.2
Embodiment 23
[0237] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 13 to 21, described by a structure:
##STR00042##
wherein
[0238] End Group D is:
##STR00043##
cMAP is
##STR00044##
[0239] Chain C is
##STR00045##
[0240] p and q are sufficient to provide a number average molecular
weight for the subunits comprising cMAP and PEG, respectively,
independently in a range of from about 500 Da to about 50,000 Da,
preferably from about 1000 Da to about 5000 Da;
[0241] m is independently at each occurrence 1, 2, 3, 4, 5, 6, 7,
8, 9, or 10, preferably 4-6 or 5;
[0242] n and r are independently at each occurrence 0, 1, 2, 3, 4,
or 5;
[0243] z is equal to or greater than 1 (up to about 2, 4, 6, 8, or
10); and
[0244] X.sub.3 and X.sub.4 are independently at each occurrence
--OH, --COOH, --C(.dbd.O)O(alkyl), --C(.dbd.O)O(aryl), --NH.sub.2,
--NH(alkyl), --N(alkyl).sub.2, or a salt or protected analog
thereof.
[0245] Again, as in Embodiments 21 and 22, to meet the molecular
weight restrictions, the numeric values for p corresponds to a
range of from about 1 to about 100, preferably from about 10 to
about 100, and the numeric values for q corresponds to as range
from about 12 to about 1200. In subsets of these embodiments, q can
also be in a range of from about 100 to about 500. In certain
subsets of this Embodiment, p and q are sufficient to provide a
number average molecular weight for the subunits comprising cMAP
and PEG, respectively, independently in a range of from about 500
Da to about 1000 Da, greater than 1000 Da to about 5000 Da, greater
than 5000 Da to about 10,000 Da, greater than 10,000 to about
25,000 Da, greater than 25,000 Da to about 50,000 Da, or any
combination of two or more of these ranges, as well as the
corresponding numeric values for these MW.sub.n ranges. In other
subsets of this Embodiment, X.sub.1 and X.sub.2 are independently
--(CH.sub.2).sub.1-4--COOH and --(CH.sub.2).sub.1-4--NH.sub.2
[0246] Additionally, or alternatively, in certain Aspects of
Embodiments 19 to 22, m is 4, 5, or 6, preferably 5.
[0247] Additionally, or alternatively, in certain Aspects of
Embodiments 19 to 22, wherein n is 1.
[0248] Additionally, or alternatively, in certain Aspects of
Embodiments 19 to 22, wherein r is 2, 3, or 4, preferably 3.
Embodiment 24
[0249] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 14 to 23, wherein p is sufficient to
provide a number average molecular weight of the subunit comprising
cMAP in a range of from about or greater than 5 kDa to about 15
kDa, from about or greater than 6 kDa to about 14 kDa, from about
or greater than 7 kDa to about 13 kDa, from about or greater than 8
kDa to about 12 kDa, from about or greater than 9 kDa to about 11
kDa, or about 10 kDa. In some subset Embodiments, for example where
the cMAP fragment has MW.sub.n of about 420 Da, this corresponds to
p having numeric values in ranges of about 12 to about 36, from
about 14 to about 33, from about 17 to about 31, from about 19 to
about 29, from about 22 to about 26, or about 24.
Embodiment 25
[0250] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 20 to 24, wherein q is sufficient to
provide a number average molecular weight of the subunit comprising
PEG in a range of from about or greater than 500 Da to about 50
kDa, from about or greater than 1 kDa to about 40 kDa, from about
or greater than 5 kDa to about 30 kDa, or from about or greater
than 5 kDa to about 20 kDa. In some of these Embodiments, for
example assuming ethylene glycol fragment has MWn of about 44 Da,
this corresponds to q having numeric values in ranges of about 11
to about 1200, from about 23 to about 910, from about 110 to about
680, or from about 110 to about 450.
Embodiment 26
[0251] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 25, wherein the linker attaching the
targeting agent to the polymer or nanoparticle core is cleavable
when subject to conditions inside an endothelial cell of the
blood-brain barrier, wherein the cleavage comprises hydrolysis,
chemical reduction, or enzymatic cleavage of the linker.
[0252] Additionally, or alternatively, the cleavable linkage
comprises one or more of an acetal, borate ester, a carboxylic
ester, a diamino, a disulfide, a ketal, a hydrazone, an imine, a
ketal, an orthoester, or a peptide linkage. Additionally, or
alternatively, the cleavable linkage further comprises a
polyethylene glycol (PEG) polymer moiety conjugated to the surface
of the nanoparticle core by a pH sensitive linkage selected from
the group consisting of nitrophenylboronic acid ester, carboxylic
acid ester, diamino ketal, orthoester, acetal, ketal, imine, and
hydrazone linkage. Additionally, or alternatively, where the
surface of the nanoparticle core is or comprises a
poly(lactic-co-glycolic acid) (PLGA) polymeror moiety, the pH
sensitive linkage may be a carboxylic acid ester linkage, wherein
the linker is dissociable when subject to pH conditions inside an
endothelial cell of the blood-brain barrier. Additionally, or
alternatively, the linker may comprise a polyethylene glycol (PEG)
polymer moiety conjugated to the surface of the nanoparticle core
by a peptide linkage, wherein the peptide linkage can be
enzymatically cleaved to cause dissociation of the ligand from the
nanoparticle when the nanoparticle is inside a brain endothelial
cell).
[0253] A variety of linkers may be used to attach the targeting
ligand to the polymer or nanoparticle core, and in some instances
may be used with different nanoparticles. In some Aspects of this
Embodiment, the cleavable linker may include a polypeptide or
chemical bond that can be chemically or enzymatically cleaved to
cause disassociation of the ligand from the nanoparticle when the
nanoparticle is inside a brain endothelial cell. For example, the
linker can incorporate an enzyme target sequence just before the
attached ligand to facilitate cleavage of the ligand following
entry into a cellular endosome, thereby separating the ligand from
the nanoparticle. In one embodiment the linker may include a
cathepsin cleavage site to promote disassociation of the ligand
from the nanoparticle. Those skilled in the art will understand
that other sequences targeted by enzymes could be employed in a
similar manner to cause disassociation of the nanoparticle from its
ligand, which will allow the nanoparticle to move into the
parenchyma of the CNS following excretion by the cell.
Alternatively, a proteasome degradation tag could also be
incorporated into the linker to cause the ligand to be degraded,
but leaving the nanoparticle itself intact, as this would
effectively dissociate the ligand and the nanoparticle following
cellular uptake. The use of linkers with particular chemical bonds
that can be chemically cleaved, such as orthoesters, acetals,
ketals, imines, and hydrazones, should also be understood to be
within the scope of this disclosure, as those skilled in the art
will appreciate that such bonds could be used to facilitate
disassociation of a ligand from a conjugated nanoparticle.
[0254] In some Aspects of this Embodiment, the cleavable linker
includes a disulfide bond that can be reduced to cause
disassociation of the ligand from the nanoparticle when the
nanoparticle is inside a brain endothelial cell. In one embodiment
the disulfide bond may be placed between two component polymers
bridging the nanoparticle and the ligand, such that upon reduction
of the disulfide bond the nanoparticle and the ligand would be
separated. In one Aspect of this Embodiment the linker is composed
of two PEG polymers that are joined by a disulfide bond where one
of the PEG polymers is conjugated to the nanoparticle core and the
other is conjugated to a ligand that mediates targeting when linked
to the nanoparticle core. After such a particle is endocytosed by a
cell, the ligand can become disassociated from the nanoparticle,
which will promote egress of the nanoparticle into the parenchyma
of the CNS.
[0255] In some Aspects of this Embodiment, the linker may include a
hydrolyzable chemical bond that can be disrupted at low pH to cause
disassociation of the ligand from the nanoparticle when the
nanoparticle is inside a brain endothelial cell. In one embodiment
the hydrolyzable bond may be placed between two component polymers
bridging the nanoparticle and the ligand, such that upon hydrolysis
of the bond the nanoparticle and the ligand would be separated. In
one embodiment the linker is composed of a PEG polymer conjugated
to the nanoparticle core at one end and linked via a diamino ketal
(DAK) to a ligand that mediates targeting. After such a particle is
endocytosed by a cell, the ligand will become disassociated from
the nanoparticle when it encounters a low pH environment, which
will promote egress of the nanoparticle into the parenchyma of the
CNS. In one Aspect, the targeting agent having a hydrolyzable
chemical bond that can be disrupted at low pH to cause
disassociation of the ligand from the nanoparticle could be formed
by having a ligand attached to a DAK linker that is attached to
PEG-orthopyridyl disulfide (OPSS), where the ligand and OPSS are at
opposite ends of the targeting agent.
[0256] In some Aspects of this Embodiment, the linker may include a
chemical bond having a pKa that can be disrupted at low pH to cause
disassociation of the ligand from the nanoparticle when the
nanoparticle is inside a brain endothelial cell. In one embodiment
the hydrolyzable bond may be placed at one end of the targeting
agent to mediate conjugation to the nanoparticle core. In this
configuration, a shift in the pH that favors hydrolysis of the bond
between the nanoparticle core and the targeting ligand would cause
the core to be separated from the ligand on the targeting ligand.
In one Aspect of this Embodiment, the targeting ligand linker
comprises a borate ester linkage, that permit covalent bonding
between the nanoparticle core and the targeting ligand, but once
the particle is endocytosed by a cell, the ligand will become
disassociated from the nanoparticle when it encounters a low pH
environment sufficient to hydrolyze the borate ester, this
hydrolysis promoting the egress of the nanoparticle into the
parenchyma of the CNS
Embodiment 27
[0257] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 26, comprising at least one borate
ester of a (nitro)phenyl boronic acid-containing polymer comprising
a structure:
##STR00046##
wherein
[0258] the polymer or nanoparticle core and the (nitro)phenyl
boronic acid-containing polymer are reversible connected to one
another by the borate condensation linkage between the
(nitro)phenyl boronic acid moieties of the (nitro)phenyl boronic
acid-containing polymer and at least one pair of vicinal diols of
the polymer or nanoparticle core, X.sub.5 being at the distal end
of this connection;
[0259] R.sup.A is nitro (or other electron withdrawing group);
[0260] n is 0, 1, 2, 3, or 4, preferably 1;
[0261] s is a number in a range of from 2 to 2000;
[0262] L is a linking group between the phenyl ring and the
polyethylene oxide linkage, the linking group comprising an amide,
carbonate, ester, or disulfide group; and
[0263] X.sub.5 is a C.sub.1-6 alkyl, optionally substituted with
--OH, --COOH, --B(OH).sub.2--, --C(.dbd.O)O(alkyl),
--C(.dbd.O)O(aryl), --NH.sub.2, --NH(alkyl), --N(alkyl).sub.2, or
--SH, and/or the at least one targeting agent coupled thereto.
[0264] In some Aspects of this Embodiment, s is any whole number
between 20 and 1200. In certain embodiments, s may be any whole
number between about 120 and about 180. In some embodiments, n may
be any whole number between about 140 and 160. In some embodiments,
s may be any one of 110, 111, 112, 113, 114, 115, 116, 117, 118,
119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131,
132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144,
145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157,
158, 159, or 160. Alternatively, the PEG segment used with any of
the linkers described herein may be about 2 kDa, about 5 kDa, about
6, kDa, about 7 kDa, about 8 kDa, about 9 kDa, about 10 kDa, about
11 kDa, about 12 kDa, about 13 kDa, about 14 kDa, or about 15
kDa.
[0265] Exemplary structures for the (nitro)phenyl boronic acid
moiety include:
##STR00047##
though it should be apparent that the borate --B(OH).sub.2
functional group can be ortho or para to the L group, as well as
meta, and nitro group can occupy any position not otherwise
occupied by the L or --B(OH).sub.2 groups.
[0266] Additionally, or alternatively, in certain Aspects of this
Embodiment, at least one of these (nitro)phenyl boronic
acid-containing polymer links or couples a single targeting agent
to the polymer or nanoparticle core. That is, while each polymer or
nanoparticle core may have a plurality of these (nitro)phenyl
boronic acid-containing polymers attached thereto, in some
independent Aspects, only one of these (nitro)phenyl boronic
acid-containing polymers (on average) links the targeting agent to
the polymer or nanoparticle core.
[0267] Additionally, or alternatively, in other Aspects of this
Embodiment, two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, or more)
of these (nitro)phenyl boronic acid-containing polymers each link
or couple independent targeting agents to a single polymer or
nanoparticle core, where the independent targeting agents may be
the same or different.
[0268] Additionally, or alternatively, in other Aspects of this
Embodiment, L is
--(C.sub.0-2alkylene-)NH--C(.dbd.O)--(C.sub.0-2alkylene)-,
--(C.sub.0-2alkylene)-C(.dbd.O)--NH--(C.sub.0-2alkylene)-,
--(C.sub.0-2alkylene)-O--C(.dbd.O)-- (C.sub.0-2alkylene)- or
--(C.sub.0-2alkylene)-C(.dbd.O)--O--(C.sub.0-2alkylene)-.
[0269] Additionally, or alternatively, in other Aspects of this
Embodiment, L is --NH--C(.dbd.O)--, --C(.dbd.O)--NH--,
--O--C(.dbd.O)--, or --C(.dbd.O)--O--.
Embodiment 28
[0270] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 27, wherein the at least one
targeting ligand independently comprises or consists of a
nucleotide, polynucleotide, aptamer, peptide, oligopeptide,
polypeptide, protein, polysaccharide, antibody or antibody
fragment.
[0271] Additionally, or alternatively, and as described elsewhere
herein, the at least one of the targeting ligands is one known to
specifically bind to receptor or surface protein expressed by a
brain endothelial cell that undergoes transcytosis. Targeting
cellular proteins that undergo transcytosis may increase the
likelihood of success of the provided methods because these
cellular proteins are known to transport proteins and other
molecules from one side of a cell to the other, often in a
coordinated manner. Additionally, or alternatively, the targeting
ligand that may be used in the compositions and with the described
methods independently comprises or consists of, but are not limited
to, transferrin, an antibody specific for the transferrin receptor,
a polypeptide that specifically binds to the transferrin receptor,
insulin, an antibody specific for the insulin receptor, a
polypeptide that specifically binds to the insulin receptor,
insulin-like growth factor 1, an antibody specific for the
insulin-like growth factor receptor 1, a polypeptide that
specifically binds to the insulin-like growth factor receptor 1,
apolipoprotein E, angiopep-2, an antibody specific for low density
lipoprotein receptor or lipoprotein receptor-related protein, a
polypeptide that specifically binds to low density lipoprotein
receptor or lipoprotein receptor-related protein; an antibody
specific for diphtheria toxin receptor, or a polypeptide that
specifically binds to diphtheria toxin receptor. Other cellular
proteins capable of facilitating transcytosis that are known in the
art may also be targeted by a ligand for carrying out the methods
disclosed herein.
[0272] Additionally, or alternatively, the targeting ligand is or
comprises any one of the foregoing targeting ligands that are known
to associate with hematopoietic differentiation antigens
(glycoproteins usually associated with CD groupings and include
CD20, CD30, CD33, and CD52). Additionally, or alternatively, the
targeting ligand is or comprises any one of the foregoing targeting
ligands that are known to associate with growth factors such as
epidermal growth factor receptor (EGFR; also known as ErbB 1),
ErbB2 (also known as HER2), ErbB3, MET, insulin-like growth factor
1 receptor (IGF 1R), ephrin receptor A3 (EphA3), TNF receptor
apoptosis-inducing ligand receptor 1 (TRAIL-R1), TRAIL-R2, and
receptor activator of nuclear factor xK3 ligand (RANKL).
Additionally, or alternatively, the targeting ligand is or
comprises any one of the foregoing targeting ligands that are known
to associate with proteins or growth factors that support the
formation of new microvasculature, including vascular endothelial
growth factor (VEGF), VEGF receptor (VEGFR), and integrins aV(33
and a5(31. Additionally, or alternatively, the targeting ligand is
or comprises any one of the foregoing targeting ligands that are
known to associate with Stromal and extracellular matrix antigens
that are therapeutic targets include fibroblast activation protein
(FAP) and tenascin.
Embodiment 29
[0273] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 28, wherein the at least one
targeting ligand is present in a range of from one to 1000,
conjugated to the polymer or nanoparticle core by the linking
groups. Additional or alternative Aspects of this Embodiment
provide independently for the presence of 1, 2, 3, 4, or 5
targeting ligands per polymer or nanoparticle core. Additional or
alternative Aspects of this Embodiment provide independently for
the presence of an average of 1, 2, 3, 4, or 5 targeting ligands
per polymer or nanoparticle core in a population of polymer or
nanoparticle core. Additionally, or alternatively, each polymer or
nanoparticle core may have attached thereto a number of targeting
ligands in a range of from 1 to 5, from 5 to 10, from 10 to 15,
from 15 to 20, from 20 to 50, from 50 to 100, from 100 to 200, from
200 to 300, from 300 to 400, from 400 to 500, or from 500 to 1000
per polymer or nanoparticle core, or a range of two or more of the
foregoing ranges, for example from 1 to 10 or from 5 to 50
targeting ligands per polymer or nanoparticle core, either on an
individual basis or on average for a population of the polymer or
nanoparticle cores.
[0274] In each case, the targeting ligands may comprise the same or
different chemical moieties suitable for this purpose.
[0275] In alternative, or additional Aspects of this Embodiment,
the polymer or nanoparticle core(s) may contain additional
cleavable linking groups that are not conjugated to the targeting
ligands (i.e., the number of cleavable linking groups attached to
the polymer or nanoparticle core is greater than the number of
conjugated targeting ligands). For example, a given polymer or
nanoparticle core may have 100 cleavable linking groups attached to
the core, only 1, 2, 3, 4, or 5 of which are conjugated to a
targeting ligand. The remaining cleavable linking groups may be
free appendages (i.e., contain a functional group capable of
binding to another chemical group, such as described elsewhere
herein) and/or may serve as a binding site for one or more of the
small or large molecule therapeutic agents or imaging agents
described herein. In such Aspects of this Embodiment, the polymer
or nanoparticle core may comprise 1, 2, 3, 4, or 5 cleavable
linking group per polymer or nanoparticle core, wherein all or only
a portion of which are conjugated to the at least one targeting
ligand. Additional or alternative Aspects of this Embodiment
provide independently for the presence of an average of 1, 2, 3, 4,
or 5 cleavable linking groups per polymer or nanoparticle core in a
population of polymer or nanoparticle core, wherein all or only a
portion of which are conjugated to the at least one targeting
ligand as set forth above. Additionally, or alternatively, each
polymer or nanoparticle core may have attached thereto a number of
cleavable linking groups in a range of from 1 to 5, from 5 to 10,
from 10 to 15, from 15 to 20, from 20 to 50, from 50 to 100, from
100 to 200, from 200 to 300, from 300 to 400, from 400 to 500, or
from 500 to 1000 per polymer or nanoparticle core, or a range of
two or more of the foregoing ranges, for example from 1 to 10 or
from 5 to 50 targeting ligands per polymer or nanoparticle core,
either on an individual basis or on average for a population of the
polymer or nanoparticle cores, wherein all or only a portion of
which are conjugated to the at least one targeting ligand as set
forth above.
[0276] In alternative, or additional Aspects of this Embodiment,
the ratio of targeting agents to cleavable linking groups attached
to the polymer or nanoparticle core may be about 100:1, about 50:1,
about 20:1, about 10:1, about 7:1, about 5:1, about 4:1, about 3:1,
about 2:1, or about 1:1. Alternatively, or alternatively, the
relative distribution of cleavable linking groups and the targeting
agents may be described in term of the percentage of total attached
conjugates that are targeting agents, relative to the total
cleavable linking groups. In some independent Aspects, the
targeting ligands are conjugated to about 1%, 2%, 3%, 4%, 5%, 6%,
7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,
95%, 98%, or substantially 100% of the cleavable linking groups
attached to the polymer or nanoparticle core. The ratios and
percentages described herein may account for mixed populations of
conjugates, such as embodiments where more than one targeting agent
and/or spacer molecule is attached to a nanoparticle core.
[0277] Additionally, the nanoparticle core and targeting ligand can
be conjugated by a linker that can facilitate disassociation of the
ligand from the nanoparticle when inside a brain endothelial cell.
In some of the described embodiments the linker may include a
disulfide bond that can be reduced to cause disassociation of the
ligand from the nanoparticle when the nanoparticle is inside a
brain endothelial cell. As also disclosed elsewhere herein, the
targeting ligand linker may include a polypeptide that can be
enzymatically cleaved to cause disassociation of the ligand from
the nanoparticle when the nanoparticle is inside a brain
endothelial cell. In some of the described embodiments the linker
may include a hydrolyzable chemical bond that can be disrupted at
low pH to cause disassociation of the ligand from the nanoparticle
when the nanoparticle is inside a brain endothelial cell. In some
of the described embodiments the linker may include a chemical bond
having a pKa that can be disrupted at low pH to cause
disassociation of the ligand from the nanoparticle when the
nanoparticle is inside a brain endothelial cell. In some of these
Aspects, the linker comprises a nitrophenyl boronic acid when
unbound to the nanoparticle that forms a nitrophenyl boronic ester
to bind to the nanoparticle core, where decoupling of the linker
and the nanoparticle core will be favored at acidic pH (e.g., about
6.8 to about 2.0). In other independent Aspects, the targeting
agent may include a diamino ketal (DAK) linkage to facilitate
disassociation of the nanoparticle and the ligand once inside a
brain endothelial cell, where decoupling of the linker and the
nanoparticle core will be favored at acidic pH. Additionally, the
described methods may be carried out using a targeting agent with a
linker having a disulfide bond that can facilitate disassociation
of an attached ligand from the nanoparticle under reducing
conditions encountered in a brain endothelial cell.
[0278] Additionally, or alternatively, while at least one targeting
ligand is linked to the polymer or nanoparticle core by a cleavable
linking group, not all targeting ligands need to be so linked. In
other Aspects of this Embodiment the polymer or nanoparticle core
may comprise a first targeting ligand and a second targeting
ligand, wherein the first targeting ligand is linked by such a
cleavable linkage, whereas the second targeting ligand is not, and
the second targeting ligand is linked by a linker that is not
amenable to disassociation from the nanoparticle core when inside
of a brain endothelial cell, and that second targeting ligand
targets the nanoparticle to a specific cell in the brain.
Embodiment 30
[0279] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 29, wherein the small molecule is a
pharmaceutical compound useful in the treatment of Alzheimer's
disease, Huntington's disease, Parkinson's disease, multiple
sclerosis, or cancer, including brain cancer, including brain
cancer metastasized from HER2-positive breast cancer.
[0280] In certain Aspects of this Embodiment, the small molecule is
a neurotransmitter. Additionally, or alternatively, the small
molecule is dopamine or serotonin.
[0281] In certain Aspects of this Embodiment, the small molecule is
a chemotherapeutic. Additionally, or alternatively, the small
molecule is one or more of Camptothecin, Erlotinib (Tarceva.RTM.),
Gefitinib (Iressa.RTM.), Imatinib (Gleevec.RTM.), Irinotecan,
Lapatinib, SN-38, or a derivative, metabolite, or prodrug
thereof.
[0282] Additionally, or alternatively, the small molecule is or
comprises abraxane, actinomycin, alitrerinoin, azacitidine,
azathioprine, bexarotene, bleomycin, bortezomib, camptothecin,
capecitabone, carboplatin, cisplatin, capecitabine, chlorambucil,
cyclophosphamide, cytarabine, dacarbazine, docetaxel,
doxifluiridine, daunorubcin, doxorubicin, epirubicin, epothilone,
erlotinib, etoposide, 5-fluorouracil, folinic acid, gefitinib,
gemcitabine, idarubicin, imatinib, irinotecan, mechlorethamine,
melphalan, mercaptopurine, methotrexate, mitoxanrone, mustine,
oxaliplatin, paclitaxel, prednisolone, procarbazine, romidepsin,
tafluposide, taxotere, teniposide, thioguainine topotecan,
tretinoin, valrubicin, vemurafenib, vinblastine, vincristine,
vindesine, vinorelbine, vismodegib, vorinosat, or a derivative,
metabolite, or prodrug thereof.
[0283] Additionally, or alternatively, the small molecule may
include molecules more broadly described in U.S. Pat. Nos.
5,747,498; 6,900,221; 7,087,613; RE41065 (corresponding to
Erlotinib); U.S. Pat. Nos. 5,457,105; 5,616,582; 5,770,599
(corresponding to Gefitinib); U.S. Pat. Nos. 6,391,874; 6,713,485;
6,727,256; 6,828,320; and 7,157,466 (corresponding to Lapatinib),
each of which is incorporated by reference herein in its entirety.
In still other Aspects of this Embodiment, additionally, or
alternatively, the small molecule may include any one of the fused
heterocyclic compounds disclosed in WO97/13771, WO98/02437,
WO00/44728, U.S. Pat. No. 6,596,878, US 2005/0148607, and US
2008/0214584; any one of the quinazoline derivatives disclosed in
WO02/02552, WO01/98277, WO003/049740, WO003/050108, and U.S. Pat.
No. 6,596,878; any one of the thienopyrimidine derivatives
disclosed in WO003/053446 and U.S. Pat. No. 7,300,935; any one of
the thienyl derivatives disclosed in U.S. Pat. No. 5,710,173; any
one of the aromatic azole derivatives disclosed in WO01/77107,
WO003/031442, U.S. Pat. Nos. 6,716,863 and 6,984,653; any one of
the bicyclic or heterocyclic aryl compounds disclosed in WO
92/20642; any one of the vinylene-azaindole derivatives disclosed
in WO94/14808; any one of the azaindoles disclosed in WO003/000688
and WO96/000226; any one of the 1-cyclopropyl-4-pyridyl-quinolones
disclosed in U.S. Pat. No. 5,330,992; any one of the styryl
compounds disclosed in U.S. Pat. Nos. 5,217,999 and 5,596,878; any
one of the styryl-substituted pyridyl compounds disclosed in U.S.
Pat. No. 5,302,606; any one of the tyrphostin-like compounds
disclosed in U.S. Pat. No. 6,225,346; any one of the seleoindoles
or selenides disclosed in WO94/03427; any one of the
1H-pyrrolo[2,3-b]pyridines disclosed in WO01/098299; any one of the
tricyclic polyhydroxylic compounds disclosed in WO92/21660; any one
of the 2-pyrazolin-5-ones disclosed in WO 01/01921), and
benzylphosphonic acid compounds disclosed in WO91/15495) Each of
these compounds have been described as compounds for use as
tyrosine kinase inhibitors for use in the treatment of cancer. Each
of these references is incorporated by reference herein in its
entirety for all purposes, or at least for the compounds disclosed
as useful in the treatment of cancer.
[0284] Alternatively, or additionally, the small molecule can also
be "tagged" with radio-isotopes for molecular imaging of the tumor
marker in vivo. Radio-labeled compounds are required for Positron
Emission Tomography (PET) and Single Photon Emission Computed
Tomography (SPECT). Such imaging enhancing agent can include at
least one of tritium, boron-10, carbon-11, gallium-68, nitrogen-13,
sulfur-35, iodine-131, or fluorine-18 at a level above the natural
abundance of each element.
Embodiment 31
[0285] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 30, wherein the small molecule
therapeutic compound is linked to the nanoparticle core by way of
an optional linker. As set forth above, the small molecule
therapeutic molecule may be chemically or electrostatically bonded
to the polymer or nanoparticle core or may be encasulated within
the structure of the nanoparticle core without being chemically or
electrostatically bonded. For example, the nanoparticle may present
a hydrophilic exterior surface in aqueous media, while having a
hydrophobic pocket within its structure, allowing for the
encapulation of a hydrophobic small molecule therapeutic to be
encapsulated within that hydrophobic pocket.
[0286] The term "optional linker" in the context of this Embodiment
refers to independent Aspects of this Embodiment where the
attachment of the small molecule therapeutic compound to the
nanoparticle core is either by a direct chemical bond or via a
linking group. The linking group may be either chemically stable
(i.e., able to maintain its structure in its presented or intended
environment) or may be cleavable by any of the mechanisms
associated with the term "cleavable" as set forth elsewhere herein.
Additionally, or alternatively, this linking group may comprise one
or more amino acid residues, such as a residue of arginine,
histidine, lysine, aspartic acid, glutamic acid, serine, threonine,
asparagine, glutamine, tyrosine, tryptophan, or a salt thereof. In
but one non-limiting example of this concept, the small molecule
therapeutic compound, camptothecin, is linked to a MAP polymer
nanopolymer surface according to:
##STR00048##
Embodiment 32
[0287] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 31, wherein the large molecule
therapeutic agent is a nucleotide, polynucleotide, aptamer,
peptide, oligopeptide, polypeptide, protein, polysaccharide,
antibody or antibody fragment useful in the treatment of
Alzheimer's disease, Huntington's disease, Parkinson's disease,
multiple sclerosi, and cancer. In certain Aspects of this
Embodiment, the large molecule is an antibody.
[0288] Additionally, or alternatively, the large molecule
therapeutic agent is an antibody and the antibody is a monoclonal
antibody. Additionally, or alternatively, the large molecule
therapeutic agent is abciximab (Reopro), adalimnumab, alemtuzumab
(Campath), basiliximab (Simulect), belirnumab (Benlysta),
bevacizumab (Avastin), bezlotoxumab (Zinplava), canakinumab
(Ilaris), certolizumab pegol (Cimzia), cetuximab (Erbitux),
daclizumab (Zenapax, Zinbryta), denosurnab (Prolia, Xgeva),
efalizurnab (Raptiva), golimumrnab (Simponi, Simrnponi Aria),
inflectra (Remicade), ipilimumab (Yervoy), ixekizumab (Taltz),
natalizumab (Tysabri), necitumumab (Portrazza), nivolumab (Opdivo),
obinutuzumab (Gazyva), ocrelizumab (Ocrevus), ofatumumab (Arzerra),
olaratumab (Lartruvo), omalizumab (Xolair), palivizumab (Synagis),
panitumumab (Vectibix), pembrolizumab (Keytruda), pertuzumab
(Perjeta), ramucirumab (Cyramza), rituximab (Rituxan), secukinumab
(Cosentyx), tocilizumab (Actemra), trastuzumab (Herceptin.RTM.),
and/or ustekinumab (Stelara). Additionally, or alternatively, the
large molecule therapeutic agent is a conjugated monoclonal
antibody. Additionally, or alternatively, the large molecule
therapeutic agent is a ibritumomab tiuxetan (Zevalin), Brentuximab
vedotin (Adcetris), or Ado-trastuzumab emtansine (Kadcyla, also
called TDM-1), denileukin diftitox (Ontak) Additionally, or
alternatively, the large molecule therapeutic agent is one or more
of ipilimumab, natalizumab, necitumumab, nivolumab, obinutuzumab,
ocrelizumab, pembrolizumab, pertuzumab, or pamucirurnab.
Additionally, or alternatively, the large molecule therapeutic
agent is one or more of adalimumab (Humira, Amjevita), Bevacizumab
(Avastin), cetuximab (Erbitux), ipilirnurnab (Yervoy), natalizumnab
(Tysabri), necitumumab (Portrazza), nivolumab (Opdivo),
obinutuzumab (Gazyva), ocrelizumrab (Ocrevus), panitumumab
(Vectibix), pembrolizumab (Keytruda), pertuzumab (Perjeta),
ramucirumab (Cyramza), rituximab (Rituxan), trastuzumab
(Herceptin). Additionally, or alternatively, the large molecule
therapeutic agent is arcitumomab, ibritumomab, capromab pendetide,
and/or tositumomab.
[0289] Additionally, or alternatively, the antibody includes one
known to associate with hematopoietic differentiation antigens
(glycoproteins usually associated with CD groupings and include
CD20, CD30, CD33, and CD52). Additionally, or alternatively, the
antibody includes one known to associate with growth factors such
as epidermal growth factor receptor (EGFR; also known as ErbB 1),
ErbB2 (also known as HER2), ErbB3, MET, insulin-like growth factor
1 receptor (IGF 1R), ephrin receptor A3 (EphA3), TNF receptor
apoptosis-inducing ligand receptor 1 (TRAIL-R1), TRAIL-R2, and
receptor activator of nuclear factor .kappa..beta. ligand (RANKL).
Additionally, or alternatively, the antibody includes one known to
associate with proteins or growth factors that support the
formation of new microvasculature, including vascular endothelial
growth factor (VEGF), VEGF receptor (VEGFR), and integrins
.alpha.V.beta.3 and .alpha.5.beta.1. Additionally, or
alternatively, the antibody includes one known to associate with
Stromal and extracellular matrix antigens that are therapeutic
targets include fibroblast activation protein (FAP) and
tenascin.
[0290] Additionally, or alternatively, the large molecule
therapeutic agent is a polynucleotide. Within this Aspect, the
polynucleotide is or comprises a genomic DNA, cDNA, mRNA, siRNA,
shRNA, miRNA, antisense oligonucleotide, virus, or a chimeric
polynucleotide. or a small molecule therapeutic agent.
Additionally, or alternatively, the polynucleotide that is an RNA
molecule, preferably an siRNA molecule.
[0291] Additionally, or alternatively, the large molecule
therapeutic agent is a fusion protein. Additionally, or
alternatively, the large molecule therapeutic agent is Abatacept,
Alefacept (Amevive)Aflibercept, or Etanercept
[0292] Additionally, or alternatively, in the context of this
Embodiment, the large molecule therapeutic compound may be attached
to the polymer or nanoparticle core either by a direct chemical
bond or via a linking group. The linking group may be either
chemically stable (i.e., able to maintain its structure in its
presented or intended environment) or may be cleavable by any of
the mechanisms associated with the term "cleavable" as set forth
elsewhere herein. Additionally, or alternatively, this linking
group may comprise one or more amino acid residues, such as for
example arginine, histidine, lysine, aspartic acid, glutamic acid,
serine, threonine, asparagine, glutamine, tyrosine, tryptophan, or
a salt thereof.
Embodiment 33
[0293] The polymer, polymer conjugate, or nanoparticle composition
of any one of Embodiments 1 to 32, wherein the composition is a
nanoparticle,
Embodiment 34
[0294] The nanoparticle of any one of claims 1 to 33, said
nanoparticle being substantially spherical and having a
cross-section dimension in a range of from about 20 nm to about 300
nm. Additional, or alternative Aspects of this Embodiment include
those where the cross-section dimension of the nanoparticles are in
a range of from 20 nm to 30 nm, from 30 nm to 40 nm, from 40 nm to
50 nm, from 50 nm to 60 nm, from 60 nm to 70 nm, from 70 nm to 80
nm, from 80 nm to 90 nm, from 90 nm to 100 nm, from 100 nm to 120
nm, from 120 nm to 140 nm, from 140 nm to 160 nm, from 160 nm to
180 nm, from 180 nm to 200 nm, from 200 nm to 220 nm, from 220 nm
to 240 nm, from 240 nm to 260 nm, from 260 nm to 280 nm, from 280
nm to 300 nm, or any combination of two or more of these foregoing
ranges, for example from 80 nm to 100 nm. In each of these cases,
the dimensions refer to measurements obtained by dynamic light
scattering, though any other method as known in the art may be
used.
Embodiment 35
[0295] The nanoparticle of any one of claims 1 to 34, the
nanoparticle having an average zeta potential of from about 0 mV to
about -15.0 mV as measured by phase analysis light scattering. In
certain Aspects of this Embodiment, the charge of the targeted
nanoparticle is near neutral. Alternatively, or additionally, the
zeta potential of the nanoparticles for use with the methods
described herein can be characterized as having a range of from
about 0 mV to about 0.5 mV, from about -0.5 mV to about -1 mV, from
about -1 mV to about -2 mV, from about -2 mV to about -3 mV, from
about -3 mV to about -4 mV, from about -4 mV to about -5 mV, from
about -5 mV to about -6 mV, from about -6 mV to about -7 mV, from
about -7 mV to about -8 mV, from about -8 mV to about -9 mV, from
about -9 mV to about -10 mV, from about -10 mV to about -11 mV,
from about -11 to about -12 mV, from about -12 mV to about -13 mV,
from about -13 mV to about -14 mV, from about -14 mV to about -15
mV, or a combination of two or more of these ranges, for example,
in a range of from about -5 mV to about -7 mV. In some Aspects of
this Embodiment the described nanoparticle will have a zeta
potential of -5.0, -5.1, -5.2, -5.3, -5.4, -5.5, -5.6, -5.7, -5.8,
-5.9, -6.0, -6.1, -6.2, -6.3, -6.4, -6.5, -6.6, -6.7, -6.8, -6.9,
-7.0, -7.1, -7.2, -7.3, -7.4, -7.5, -7.6, -7.7, -7.8, -7.9, or -8.0
mV.
Embodiment 36
[0296] The nanoparticle of any one of Embodiments 1 to 35, the
nanoparticle further comprising an imaging agent. In certain
Aspects of this Embodiment, the imaging agent is present in
addition to the small molecule or large molecule therapeutic. In
other independent Aspects of this Embodiment, the imaging agent is
present in lieu of one or both of the small molecule or large
molecule therapeutic. In certain Aspects of this Embodiment, the
imaging agent is Cu-64.
Embodiment 37
[0297] A plurality of nanoparticles, each individual nanoparticle
comprising the composition of any one of Embodiments 1 to 32 or
described in terms of any one of Embodiments 33 to 36.
Embodiment 38
[0298] The plurality of nanoparticles of Embodiment 36, each
individual nanoparticle being described by the composition of any
one of Embodiments 1 to 32 or the characteristics of any one of
Embodiments 33 to 36, the plurality of nanoparticles being
substantially monodispersed, exhibiting a standard deviation in
cross-sectional dimension (i.e., diameter) among the nanoparticles
of less than 20%, 30%, 40%, 50%, or 60%, as measured by
cryo-transmission electron microscopy (cryo-TEM).
Embodiment 39
[0299] A pharmaceutical composition comprising the polymer, polymer
conjugate, or nanoparticle composition of any one of Embodiments 1
to 38, the composition comprising a biologically active agent and
pharmaceutically acceptable carrier or excipient.
[0300] In certain Aspects of this Embodiment, the polymers,
polymer-conjugates, and/or nanoparticles may exist as dispersions
in aqueous media, said aqueous media optionally also containing
buffers, surfactants, or other modifiers. The present disclosure
also contemplates pharmaceutical compositions comprising one or
more biologically active agents and any of the polymers or polymer
conjugates or nanoparticles or plurality of nanoparticles described
herein and a pharmaceutically acceptable vehicle, carrier or
excipient.
Embodiment 40
[0301] A method of delivering enhanced levels of a therapeutic
agent or imaging agent to a brain parenchyma of a subject having a
neurological brain disorder, the method comprising systemically
administering the nanoparticles of any one of Embodiments 1 to 38
or a pharmaceutical composition of Embodiment 39 to a patient in
need of such enhanced delivery. Implicit in certain aspects of this
description is that at least one of the therapeutic agents or
imaging agents being ferried by the nanoparticle composition may
itself be unable to make the passage across the blood brain barrier
to achieve therapeutically effective amounts in the parenchyma
without the use of the nanoparticles disclosed herein (i.e., the
cargo falls outside the definition of small molecule capable of
passing the blood brain barrier set forth elsewhere herein). In
this regard, the bioavailability of the therapeutic or imaging
agent is improved using the nanoparticle, relative to an
administration of the therapeutic or imaging agent by itself. The
description of enhanced delivery or enhanced levels is intended to
reflect the ability of the disclosed nanoparticles to pass through
the BBB, by transcytosis or other means, to deliver the cargo
(therapeutic and/or imaging agents) at levels and/or rates higher
than the cargo can be delivered themselves (without being ferried
by the nanoparticles).
[0302] Additionally, or alternatively, further Aspects of this
Embodiment comprise method of treating a neurological disorder in a
patient, the methods comprising systemically administering a first
small molecule therapeutic agent and/or a large molecule
therapeutic agent to the patient in need of such treatment,
wherein
[0303] (a) the first small molecule therapeutic agent and/or the
large molecule therapeutic agent is attached to a nanoparticle
comprising a nanoparticle core and a targeting agent, the targeting
agent comprising at least one targeting ligand attached to an
external surface of the nanoparticle core by a linker having a
cleavable linkage; [0304] (i) the external surface of the
nanoparticle comprising a neutral and/or negatively charged mucic
acid-containing polymer (MAP) (including substantially free of
cationic mucic-acid containing polymer (cMAP)); [0305] (ii) the at
least one targeting ligand having an affinity for binding to a
receptor expressed by endothelial cells of the blood-brain barrier;
and [0306] (iii) the cleavable linkage being cleavable when subject
to conditions inside an endothelial cell of the blood-brain
barrier, wherein the cleavage comprises hydrolysis, chemical
reduction, or enzymatic cleavage of the linker; and wherein one or
both of [0307] (iv) the small molecule therapeutic agent is
optionally linked to the nanoparticle core by way of an optional
linker; and/or [0308] (v) the large molecule therapeutic agent is
linked to the nanoparticle by way of an optional linker; and
[0309] (b) the administration of the first small molecule
therapeutic agent and/or the large molecule therapeutic agent
attached to the nanoparticle results in the delivery of the first
small molecule therapeutic agent and/or the large molecule
therapeutic agent past the blood brain barrier and into the
subject's brain parenchyma in an amount is greater than would be
delivered were the first small molecule therapeutic agent and/or
the large molecule therapeutic agent not attached to the
nanoparticle.
[0310] Additionally, or alternatively, in some further Aspects of
this Embodiment, a useful figure of merit for measuring the utility
of these methods is an enhancement ratio of the steady state amount
of small or large molecule therapeutic or imaging agent present in
the brain parenchyma, having passed through the blood brain barrier
after systemic delivery through use of these nanoparticles,
relative to the amount of small or large molecule therapeutic or
imaging agent present in the brain parenchyma, having passed
through the blood brain barrier after systemic delivery of the
small or large molecule therapeutic or imaging agent by itself,
under otherwise comparable conditions. In certain Aspects of this
Embodiment, this enhancement ratio can range from 2 to 3 times,
from 3 to 5 times, from 5 to 10 times, from 10 to 25 times, from 25
to 50 times, from 50 to 100 times, from 100 time to 500 times, or a
combination of two or more of these foregoing ranges, depending on
the nature of the therapeutic or imaging agent.
[0311] Additionally, or alternatively, the enhanced level of the
therapeutic agent delivered by the nanoparticles to the brain
parenchyma can be defined as an amount that is greater than is
delivered using otherwise equivalent nanoparticles that do not
contain the cleavable linker under the same conditions.
Additionally, or alternatively, the method may be characterized as
a method of delivering enhanced levels of a therapeutic agent to a
brain parenchyma of a subject having a neurological brain disorder,
said therapeutic agent being unable to achieve therapeutically
effective levels in the brain parenchyma through systemic
administration of the agent itself, the method comprising
systemically administering a plurality of one or more of the
nanoparticles described herein to the subject having the
neurological brain disorder (including any of the diseases set
forth elsewhere herein) and in need of delivery of an enhanced
level of the therapeutic agent across a blood-brain barrier to the
subject's brain parenchyma, the plurality of nanoparticles being
administered at a dose rate sufficient to enhance the delivery of
the chemotherapeutic agent to the brain parenchyma in
therapeutically useful quantities that are higher than available by
systematically administering the free therapeutic agent alone.
[0312] Additionally, or alternatively, the methods of treating a
patient/subject having a neurological condition may include use of
the disclosed nanoparticles in combination with conventional drug
or other therapy. For example, while the nanoparticle compositions
described herein have been described as especially useful for
ferrying therapeutics and imaging agents (that are otherwise
blocked to any practical extent), across the blood brain barrier,
these disclosed nanoparticle compositions may also be used to
complement conventional treatments involving the use of
therapeutics that can, by themselves, pass the blood brain barrier.
While the number of small molecule therapeutics capable of passing
through the blood brain barrier is relatively small (e.g., less
than 2% of small molecule therapeutics and essentially 100% of
large-molecule therapeutics), some can make this passage, as
described elsewhere herein.
[0313] Accordingly, independent Aspects of this Embodiment include
those methods of treating a patient known or believed to have a
neurological disorder, the method comprising systemically
administering a composition described in any one of Embodiments 1
to 39 to a patient or subject in need of such treatment, wherein in
the composition comprises at least one small and/or large molecule
therapeutic agent and/or imaging agent that by itself is incapable
of passing through the blood brain barrier in sufficient amounts to
yield therapeutically effective amounts in the brain pa