U.S. patent application number 17/695230 was filed with the patent office on 2022-07-07 for paclitaxel-albumin-binding agent compositions and methods for using and making the same.
The applicant listed for this patent is Mayo Foundation for Medical Education and Research. Invention is credited to Svetomir N. Markovic, Wendy K. Nevala.
Application Number | 20220211870 17/695230 |
Document ID | / |
Family ID | 1000006196517 |
Filed Date | 2022-07-07 |
United States Patent
Application |
20220211870 |
Kind Code |
A1 |
Markovic; Svetomir N. ; et
al. |
July 7, 2022 |
PACLITAXEL-ALBUMIN-BINDING AGENT COMPOSITIONS AND METHODS FOR USING
AND MAKING THE SAME
Abstract
Described herein are nanoparticle compositions comprising
binding agents, carrier proteins and an, amount of paclitaxel
derivative, and optionally a therapeutic agent. Also described
herein are nanoparticle compositions comprising carrier proteins
and an amount of paclitaxel, and optionally binding agents and/or a
therapeutic agent, wherein the paclitaxel is present in an amount
that is less than an amount that provides a therapeutic effect.
Also disclosed herein are nanoparticles which contain (a) carrier
protein, (b) a paclitaxel derivative, the paclitaxel derivative
haying reduced toxicity compared to paclitaxel, and optionally (c)
a binding agent and/or (d) a therapeutic agent. Also described are
methods of making and using the same, in particular, as a cancer
therapeutic.
Inventors: |
Markovic; Svetomir N.;
(Rochester, MN) ; Nevala; Wendy K.; (Rochester,
MN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Mayo Foundation for Medical Education and Research |
Rochester |
MN |
US |
|
|
Family ID: |
1000006196517 |
Appl. No.: |
17/695230 |
Filed: |
March 15, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16328146 |
Feb 25, 2019 |
11311631 |
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PCT/US2017/050355 |
Sep 6, 2017 |
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17695230 |
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62384119 |
Sep 6, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 47/6803 20170801; A61K 47/6851 20170801; A61K 47/643 20170801;
A61K 47/54 20170801; A61K 31/337 20130101; A61K 47/6929
20170801 |
International
Class: |
A61K 47/69 20060101
A61K047/69; A61K 47/64 20060101 A61K047/64; A61K 47/54 20060101
A61K047/54; A61K 47/68 20060101 A61K047/68; A61P 35/00 20060101
A61P035/00; A61K 31/337 20060101 A61K031/337 |
Claims
1-90. (canceled)
91. A nanoparticle complex comprising albumin and paclitaxel,
wherein the paclitaxel is present in an amount that is less than an
amount that provides a therapeutic effect.
92. The nanoparticle complex of claim 91, wherein the albumin and
the paclitaxel have a relative weight ratio of greater than about
10:1, or about 11:1, or about 12:1, or about 13:1, or about 14:1,
or about 15:1, or about 16:1, or about 17:1, or about 18:1, or
about 19:1, or about 20:1, or about 21:1, or about 22:1, or about
23:1, or about 24:1, or about 25:1, or about 26:1, or about 27:1,
or about 28:1, or about 29:1, about 30:1, about 31:1, about 32:1,
about 33:1, about 34:1, about 35:1 or about 40:1 in the
nanoparticle complex.
93. The nanoparticle complex of claim 91, wherein the amount of
paclitaxel present in the nanoparticle composition is less than
about 4.54 mg/mL, or about 4.16 mg/mL, or about 3.57 mg/mL, or
about 3.33 mg/mL, or about 3.12 mg/mL, or about 2.94 mg/mL, or
about 2.78 mg/mL, or about 2.63 mg/mL, or about 2.5 mg/mL, or about
2.38 mg/mL, or about 2.27 mg/mL, or about 2.17 mg/mL, or about 2.08
mg/mL, or about 2 mg/mL, or about 1.92 mg/mL, or about 1.85 mg/mL,
or about 1.78 mg/mL, or about 1.72 mg/mL, or about 1.67 mg/mL.
94. The nanoparticle complex of claim 91, further comprising
antibodies associated with the nanoparticle complex.
95. The nanoparticle complex of claim 94, wherein the antibodies
are selected from ado-trastuzumab emtansine, alemtuzumab,
atezolizumab, bevacizumab, cetuximab, denosumab, dinutuximab,
ipilimumab, nivolumab, obinutuzumab, ofatumumab, panitumumab,
pembrolizumab, pertuzumab, rituximab, avelumab, durvalumab,
pidilizumab, BMS 936559, OKT3, and trastuzumab.
96. The nanoparticle complex of claim 91, wherein the albumin is
human serum albumin.
97. The nanoparticle complex of claim 96, wherein the human serum
albumin is recombinant human serum albumin.
98. The nanoparticle complex of claim 91, wherein the paclitaxel is
present in an amount effective for providing stability to the
nanoparticles.
99. The nanoparticle complex of claim 91, wherein the nanoparticle
complex is held together by non-covalent bonds between the albumin
and the paclitaxel.
100. The nanoparticle complex of claim 91, further comprising a
therapeutic agent.
101. The nanoparticle complex of claim 100, wherein the therapeutic
agent is selected from abiraterone, bendamustine, bortezomib,
carboplatin, cabazitaxel, cisplatin, chlorambucil, dasatinib,
docetaxel, doxorubicin, epirubicin, erlotinib, etoposide,
everolimus, gefitinib, idarubicin, imatinib, hydroxyurea, imatinib,
lapatinib, leuprorelin, melphalan, methotrexate, mitoxantrone,
nedaplatin, nilotinib, oxaliplatin, paclitaxel, pazopanib,
pemetrexed, picoplatin, romidepsin, satraplatin, sorafenib,
vemurafenib, sunitinib, teniposide, triplatin, vinblastine,
vinorelbine, vincristine, or cyclophosphamide.
102. The nanoparticle complex of claim 100, wherein the paclitaxel
is present in an amount effective for providing affinity of the at
least one therapeutic agent to the albumin.
103. The nanoparticle complex of claim 100, wherein the paclitaxel
is present in an amount effective for facilitating complex
formation of the at least one therapeutic agent and the
albumin.
104. A nanoparticle composition comprising the nanoparticle complex
of claim 91.
105. The nanoparticle composition of claim 104 that is
lyophilized.
106. A method for treating cancer in a patient in need thereof, the
method comprising administering a nanoparticle complex of claim 91
or a nanoparticle composition of claim 104 to a patient having
cancer.
107. A method for forming an albumin-paclitaxel nanoparticle,
wherein the method comprises: homogenizing the albumin with
paclitaxel in a solution under high pressure, to generate the
albumin-paclitaxel nanoparticle, wherein the ratio of albumin to
paclitaxel is greater than 10:1.
108. The method of claim 107, further comprising contacting the
albumin-paclitaxel nanoparticle with an antibody.
Description
FIELD OF THE INVENTION
[0001] This application relates to novel compositions of binding
agents and carrier proteins and methods of making and using the
same, in particular, as a cancer therapeutic.
BACKGROUND
[0002] Chemotherapy remains a mainstay for systemic therapy for
many types of cancer, including melanoma. Most chemotherapeutic
agents are only slightly selective to tumor cells, and toxicity to
healthy proliferating cells can be high (Allen T M. (2002) Cancer
2: 750-763), often requiring dose reduction and even
discontinuation of treatment. In theory, one way to overcome
chemotherapy toxicity issues as well as improve drug efficacy is to
target the chemotherapy drug to the tumor using antibodies that are
specific for proteins selectively expressed (or overexpressed) by
cancer cells to attract targeted drugs to the tumor, thereby
altering the biodistribution of the chemotherapy and resulting in
more drug going to the tumor and less affecting healthy tissue.
Despite 30 years of research, however, specific targeting rarely
succeeds in the therapeutic context.
[0003] Conventional antibody dependent chemotherapy (ADC) is
designed with a toxic agent linked to a targeting antibody via a
synthetic protease-cleavable linker. The efficacy of such ADC
therapy is dependent on the ability of the target cell to bind to
the antibody, the linker to be cleaved, and the uptake of the toxic
agent into the target cell. Schrama, D. et ai, (2006) Nature
reviews. Drug discovery 5:147-159.
[0004] Antibody-targeted chemotherapy promised advantages over
conventional therapy because it provides combinations of targeting
ability., multiple cytotoxic agents, and improved therapeutic
capacity with potentially less toxicity. Despite extensive
research, clinically effective antibody-targeted chemotherapy
remains elusive: major hurdles include the instability of the
linkers between the antibody and chemotherapy drug, reduced tumor
toxicity of the chemotherapeutic agent when bound to the antibody,
and the inability of the conjugate to bind and enter tumor cells.
In addition, these therapies did not allow for control over the
size of the antibody-drug conjugates.
[0005] There remains a need in the art for antibody-based cancer
therapeutics that retain cytotoxic effect for targeted drug
delivery to provide reliable and improved anti-tumor efficacy over
prior therapeutics.
SUMMARY
[0006] According to the the present invention are disclosed
nanoparticles which contain (a) carrier protein, (b) optionally a
binding agent, and (c) paclitaxel, wherein the paclitaxel is
present in an amount that is less than an amount that provides a
therapeutic effect, and optionally (d) a therapeutic agent. Some
aspects of the current invention are predicated, in part, on the
idea that a reduced amount of paclitaxel, for example compared to
that in albumin-bound nanoparticles such as ABRAXANE.RTM.,
facilitates the formation of a complex of a carrier protein, such
as albumin, with a binding agent, such as an antibody, to provide a
stable nanoparticle. These nanoparticles may be referred to herein
as "reduced toxicity nanoparticles" (RTP) or RTP complexes.
[0007] Also disclosed herein are nanoparticles which contain (a)
carrier protein (b) a paclitaxel derivative, the paclitaxel
derivative having reduced toxicity compared to paclitaxel, and
optionally (c) a binding agent and/or (d) a therapeutic agent.
[0008] Further described herein are nanoparticle compositions, and
methods of making and using the nanoparticles.
[0009] In one aspect is provided a nanoparticle comprising albumin
and a paclitaxel derivative, wherein the paclitaxel derivative is
less toxic than paclitaxel. In one embodiment, the paclitaxel
derivative is a Meerwein Product of Paclitaxel. In a preferred
embodiment, the paclitaxel derivative is
20-acetoxy-4-deactyl-5-epi-20, O-secotaxol. In one embodiment, the
paclitaxel derivative is Baccatin III (2.beta., 5.alpha., 7.alpha.,
10.alpha.,
13.beta.)-4,10-Diacetoxy-1,7,13-trihydroxy-9-oxo-5,20-epoxytax-11-en-2-yl
benzoate). In one embodiment, the nanoparticle is held together by
non-covalent bonds between the albumin and the paclitaxel
derivative. In one embodiment, the albumin and the paclitaxel
derivative have a relative weight ratio of less than about 10:1. In
one embodiment, the nanoparticles do not comprise paclitaxel.
[0010] In one embodiment is provided a nanoparticle complex
comprising albumin and a paclitaxel derivative, and further
comprises binding agents (e.g., antibodies) having an
antigen-binding domain. In one embodiment, an amount of the binding
agents (e.g., antibodies) are arranged on an outside surface of the
nanoparticle complexes. In some embodiments, the binding agents are
a substantially single layer of binding agents on all or part of
the surface of the nanoparticle. In one embodiment, the binding
agents are bound to the carrier protein by non-covalent bonds.
Preferably, the binding agents are antibodies.
[0011] In one embodiment, the nanoparticle complex further
comprises a therapeutic agent. In one embodiment, die therapeutic
agent is abiraterone, bendamustine, bortezomib, carboplatin,
cabazitaxel, cisplatin, chlorambucil, dasatinib, docetaxel,
doxorubicin, epirubicin, erlotinib, etoposide, everolimus,
gefitinib, idarubicin, imatinib, hydroxyurea, irnatinib, lapatinib,
icuprorelin, melphalan, methotrexate, mitoxantrone, nedaplatin,
nilotinib, oxaliplatin, paclitaxel, pazopanib, pemetrexed,
picoplatin, romidepsin, satraplatin, sorafenib, vemurafenib,
sunitinib, teniposide, triplatin, vinblastine, vinorelbine,
vincristine, or cyclophosphamide. In one embodiment, the
therapeutic agent is an agent listed in Table 2.
[0012] In one aspect is provided a nanoparticle composition
comprising the nanoparticles or nanoparticle complexes as described
herein.
[0013] In one aspect is provided a nanoparticle comprising a
carrier protein (e.g., albumin) and paclitaxel, wherein the
paclitaxel is is present in an amount that is less than an amount
that provides a therapeutic effect. In one embodiment, the
nanoparticle is held together by non-covalent bonds between the
carrier protein and the paclitaxel. In one aspect, the nanoparticle
complex was made by combining the carrier protein (e.g., albumin)
and paclitaxel at a relative weight ratio of greater than about
10:1 carrier protein to paclitaxel.
[0014] In one embodiment, the amount of paclitaxel present in the
nanoparticles (nanoparticle complexes) or nanoparticle composition
is greater than or equal to a minimum amount capable of providing
stability to the nanoparticle complexes comprising a protein
carrier (e.g., albumin) and paclitaxel. In one embodiment, the
amount of paclitaxel present in the nanoparticles or nanoparticle
composition is greater than or equal to a minimum amount capable of
providing affinity of the paclitaxel with the protein carrier
(e.g., albumin). In one embodiment, the amount of paclitaxel
present in the nanoparticles or nanoparticle composition is greater
than or equal to a minimum amount capable of facilitating complex
formation of the paclitaxel and the protein carrier (e.g.,
albumin).
[0015] The ratio of albumin to paclitaxel in the nanoparticles or
nanoparticle composition preferably is less than that present in
ABRAXANE.RTM.. ABRAXANE.RTM. contains 100 mg paclitaxel for about
900 mg albumin. In one embodiment, the weight ratio of the carrier
protein (e.g., albumin) to paclitaxel in the nanoparticle
composition is greater than about 9:1. In one embodiment, the
weight ratio is greater than about 10:1, or 11:1, or 12:1, or 13:1,
or 14:1, or 15:1, or about 16:1, or about 17:1, or about 18:1, or
about 19:1, or about 20:1, or about 21:1, or about 22:1, or about
23:1, or about 24:1, or about 25:1, or about 26:1, or about 27:1,
or about 28:1, or about 29:1, or about 30:1, in one embodiment, the
weight ratio of the carrier protein to paclitaxel in the
nanoparticle complex is greater than about 9:1. In one embodiment,
the weight ratio is greater than about 10:1, or 11:1, or 12:1, or
13:1, or 14:1, or 15:1, or about 16:1, or about 17:1, or about
18:1, or about 19:1, or about 20:1, or about 21:1, or about 22:1,
or about 23:1, or about 24:1, or about 25:1, or about 26:1, or
about 27:1, or about 28:1, or about 29:1, or about 30:1.
[0016] In one embodiment, the amount of paclitaxel is greater than
or equal to a minimum amount capable of providing stability to the
nanoparticle complexes comprising a protein carrier (e.g., albumin)
and paclitaxel, and optionally at least one therapeutic agent. In
one embodiment, the amount of paclitaxel is greater than or equal
to a minimum amount capable of providing affinity of the at least
one therapeutic agent to the protein carrier. In one embodiment,
the amount of paclitaxel is greater than or equal to a minimum
amount capable of facilitating complex formation of the at least
one therapeutic agent and the protein carrier.
[0017] In any of the embodiments, the amount of paclitaxel can be
less than a therapeutic amount for paclitaxel. In other words, the
amount can be less than what is provided or contemplated for
providing a therapeutic benefit, such as for example, a
chemotherapeutic amount to effectively treat a cancer.
[0018] In one embodiment, the amount of paclitaxel present in the
nanoparticle composition is less than about 5 mg/mL. In one
embodiment, the amount of paclitaxel present in the nanoparticle
composition is less than about 4.54 mg/mL, or about 4.16 mg/mL, or
about 3.57 mg/mL, or about 3.33 mg/mL, or about 3.12 mg/mL, or
about 2.94 mg/mL, or about 2.78 mg/mL, or about 2.63 mg/mL, or
about 2.5 mg/mL, or about 2.38 mg/mL, or about 2.27 mg/mL, or about
2.17 mg/mL, or about 2.08 mg/mL, or about 2 mg/mL or about 1.92
mg/mL, or about 1.85 mg/mL, or about 1.78 mg/mL, or about 1.72
mg/mL, or about 1.67
[0019] Without being bound by theory, it is contemplated that
binding to the carrier protein, e.g., complexation of the binding
agent to the carrier protein, occurs through an albumin-binding
motif on the binding agents and/or an antibody-binding motif on the
carrier protein, In one embodiment, the binding agent comprises an
albumin-binding motif. In one embodiment, the carrier protein
comprises an antibody-binding motif. Non-limiting examples of
antibody-binding motifs can be found in PCT Application No,
PCT/US2017/045643, filed Aug. 4, 2017, which is incorporated herein
by reference in its entirety. In some embodiments, the binding
agent is a non-therapeutic and non-endogenous human antibody, a
fusion protein, e.g., fusion of an antibody Fc domain to a peptide
that binds a target antigen, or an aptamer.
[0020] In one embodiment, the binding agent comprises an
antigen-binding domain. In one embodiment, the antigen is CD3,
CD19, CD20, CD38, CD30, CD33, CD52, PD-1, PD-L1, PD-L2, CTLA-4,
RANK-L, GD-2, Ly6E, HER3, EGFR, DAF, ERBB-3 receptor, CSF-1R, HER2,
STEAP1, CD3, CEA, CD40, OX40, Ang2-VEGF, or VEGF.
[0021] Tin one embodiment, the binding agent is an antibody
selected from ado-trastuzumab emtansine, alemtuzumab, atezolizumab,
bevacizuniab, cetuximab, denosumab, dinutuximab, ipilimumab,
nivolumab, obinutuzumab, ofatumumab, panitumurnab, pembrolizuniab,
pertuzumab, rituximab, Ramucirumab, avelumab or durvaiumab,
pidilizumab, BMS 936559, OKT3, and trastuzumab.
[0022] The invention further includes lyophilized nanoparticles and
nanoparticle compositions, and lyophilized nanoparticles and
compositions that do not materially differ from, or are the same
as, the properties of freshly-prepared nanoparticles. In
particular, the lypholized composition, upon resuspending in
aqueous solution, is similar or identical to the fresh composition
in terms of particle size, particle size distribution, toxicity for
cancer cells, binding agent affinity, and binding agent
specificity, Surprisingly, lyophilized nanoparticles retain the
properties of freshly-made nanoparticles after resuspension,
notwithstanding the presence of two different protein components in
these particles. In one embodinlent, the lyophilized composition is
stable at room temperature for at least about 3 months, 4 months, 5
months, 6 months, 7 months, 8 months, 9 months, 10 months, 11
months, 12 months, or longer. in one embodiment, the lyophilized
composition is stable at room temperature for at least 3 months. In
one embodiment, the reconstituted nanoparticles retain the activity
of the therapeutic agent and are capable of binding to the target
in vivo.
[0023] In some embodiments, the at least one therapeutic agent is
located inside the nanoparticle. In other embodiments, the at least
one therapeutic agent is located on the outside surface of the
nanoparticle. In yet other embodiments, the at least one
therapeutic agent is located inside the nanoparticle and on the
outside surface of the nanoparticle.
[0024] In some embodiments, the nanoparticle contains more than one
type of therapeutic agent. For example, a taxane and a platinum
drug, e.g. paclitaxel and cisplatin.
[0025] In some embodiments, the nanoparticle further comprises at
least one additional therapeutic agent that is not paclitaxel. In
some embodiments, the at least one therapeutic agent is
abiraterone, bendaniustine, bortezomib, carboplatin, cabazitaxel,
cisplatin, chlorambucil, dasatinib, docetaxel, doxorubicin,
epirubicin, erlotinib, etoposide, eyerolimus, gefitinib,
idarubicin, imatinib, hydroxyurea, imatinib, lapatinib,
leuprorelin, melphalan, methotrexate, mitoxantrone, nedaplatin,
nilotinib, oxaliplatin, pazopanib, pemetrexed, picoplatin,
romidepsin, satraplatin, sorafenib, yernuralenib, sunitinib,
teniposide, triplatin, vinblastine, vinorelbine, vincristine, or
cyclophosphamide.
[0026] In some embodiments, the binding agents, carrier protein
and/or, when present, therapeutic agent, are bound through
non-covalent bonds.
[0027] In some embodiments, the carrier protein is selected from
the group consisting of gelatin, elastin gliadin, legumin, zein, a
soy protein, a milk protein, and a whey protein. In preferred
embodiments, the carrier protein is albumin, for example, human
serum albumin.
[0028] In some embodiments, the composition is formulated for
intravenous delivery. In other embodiments, the composition is
formulated for direct injection or perfusion into a tumor.
[0029] In some embodiments, the nanoparticles have a dissociation
constant between about 1.times.10.sup.-11 M and about
1.times.10.sup.-9M.
[0030] In one embodiment, provided herein are methods of making the
nanoparticle compositions, wherein said method comprises contacting
the carrier protein and the paclitaxel and the at least one
therapeutic agent under conditions and ratios of components that
will allow for formation of the desired nanoparticles.
[0031] In some aspects, provided herein are methods for forming an
albumin-paclitaxel derivative or albumin-paclitaxel nanoparticle,
wherein method comprises: homogenizing the albumin with paclitaxel
derivative or paclitaxel in a solution under high pressure, to
generate an albumin-paclitaxel derivative or albumin-paclitaxel
nanoparticle.
[0032] In some aspects, provided herein are methods of making
nanoparticle compositions, wherein said methods comprise contacting
the carrier protein, the paclitaxel, and/or the therapeutic agent
with the binding agents in a solution having a pH of between 5.0
and 7.5 and a temperature between about 5.degree. C. and about
60.degree. C., between about 23.degree. C. and about 60.degree. C.,
or between about 55.degree. C. and about 60.degree. C. under
conditions and ratios of components that will allow for formation
of the desired nanoparticles. In one embodiment, the nanoparticle
is made between 55.degree. C. and 60.degree. C. and pH 7.0. In
another aspect, provided herein are methods of making the
nanoparticle compositions, wherein said method comprises (a)
contacting the carrier protein, the paclitaxel and the therapeutic
agent to form a core and (b) optionally contacting the core with
the antibodies in a solution having a pH of about 5.0 to about 7.5
at a temperature between about 5.degree. C. and about 60.degree.
C., between about 23.degree. C. arid about 60.degree. C., or
between about 55.degree. C. and about 60.degree. C. under
conditions and ratios of components that will allow for formation
of the desired nanoparticles.
[0033] In some aspects, an amount of a therapeutic agent (e.g., a
therapeutic agent which is not paclitaxel) can also be added to the
carrier protein.
[0034] In further embodiments, the nanoparticles are made as above,
and then lyophilized.
[0035] In another aspect, provided herein are methods for treating
a cancer cell, the method comprising contacting the cell with an
effective amount of a nanoparticle composition disclosed herein to
treat the cancer cell.
[0036] In one embodiment is provided a method for killing cancer
cells in a population of cancer cells, the method comprising
contacting the cells with an effective amount of a nanoparticle
composition, wherein said composition is maintained in contact with
said cells for a sufficient period of time to kill cancer cells,
wherein said nanoparticle composition comprises nanoparticle
complexes, each of the nanoparticles comprising albumin and
paditaxel, wherein the paclitaxel is present in an amount that is
less than an amount that provides a therapeutic effect.
[0037] In one embodiment is provided a method for treating cancer
in patient in need thereof, the method comprising administering to
the patient a nanoparticle composition comprising nanoparticle
complexes, each of the nanoparticles comprising albumin and
paclitaxekwherein the paclitaxel is present in an amount that is
less than an amount that provides a therapeutic effect.
[0038] In another aspect, provided herein are methods for treating
a tumor in a patient in need thereof, the method comprising
contacting the cell with an effective amount of a nanoparticle
composition disclosed herein to treat the tumor. In some
embodiments, the size of the tumor is reduced. In other
embodiments, the nanoparticle composition is administered
intravenously. In yet other embodiments, the nanoparticle
composition is administered by direct injection or perfusion into
the tumor.
[0039] In some embodiments, the methods provided herein include the
steps of: a) administering the nanoparticle composition once a week
for three weeks; b) ceasing administration of the nanoparticle
composition for one week; and c) repeating steps a) and b) as
necessary to treat the tumor.
[0040] In some embodiments, the therapeutically effective amount
comprises about 75 mg/m.sup.2 to about 175 mg/m.sup.2 of the
carrier protein (i.e., milligrams carrier protein per m.sup.2 of
the patient). In other embodiments, the paclitaxel is of an amount
that is less than about 75 mg/m.sup.2, such as between 5 mg/m.sup.2
and 75 mg/m.sup.2. In other embodiments, the paclitaxel is of an
amount that is less than a therapeutically effective amount. In
some embodiments, the therapeutic agent is of a therapeutically
effective amount. In some embodiments, the therapeutically
effective amount comprises about 30 mg/m.sup.2 to about 70
mg/m.sup.2of the binding. agent. In yet other embodiments, the
therapeutically effective amount comprises about 30 mg/m.sup.2 to
about 70 mg/m.sup.2bevacizumab.
[0041] An embodiment of the invention includes a method for
increasing the duration of tumor uptake of a chemotherapeutic agent
by administering the chemotherapeutic agent in a nanoparticle
comprising a carrier protein and the chemotherapeutic agent having
surface complexation with an antibody, e,g., an antibody that
specifically binds to an antigen on or shed by the tumor.
BRIEF DESCRIPTION OF TFIE DRAWINGS
[0042] The following figures are representative only of the
invention and are not intended as a limitation. For the sake of
consistency, the nanoparticles of this invention using
ABRAXANE.RTM. and bevacizumab employ the acronym "AR" and the
number after AB such as AB160 is meant to confer the average
particle size of these nanoparticles (in nanometers). Likewise,
when the binding agent is rituximab, the acronym is "AR" while the
number thereafter remains the same.
[0043] FIG. 1 shows the reaction of paclitaxel withi Meetwein's
Reagent to form the Meerwein's Product of Paclitaxel
(20-Acetoxy-4-deactyl-5-epi-20, O-secotaxol).
[0044] FIGS. 2A-2E show the nanoparticle diameter of non-toxic
nanoparticles (NTP) alone (FIG. 2A) or incubated with 4 mg/mL (FIG.
2B), 6 mg/mL (FIG. 2C), 8 mg/mL (FIG. 2D) or 10 mg/mL (FIG. 2E)
bevacizumab for 30 min. Diameter was measured at a 1:300 dilution
using Malvern Nanosight technology.
[0045] FIGS. 3A and 3B show the binding affinity (Kd) of
bevacizumab (FIG. 3A) or rituximab (FIG. 3B) to NTP. Kd was
measured using Bio Layer Interferometry Technology.
[0046] FIG. 4A shows the stability of nanoparticle complexes in
PBS. FIG. 4B shows the stability of nanoparticle complexes in
serum.
[0047] FIGS. 5A-5E show blocking of CD20-positive Daudi cells with
isotype control (FIG. 5A), anti-CD20 antibody (FIG. 5B),
ABPAXANE.RTM. (FIG. 5C), ARI60 (FIG. 5D), NTP (FIG. 5E),
rituxitmab-bound NTP (NTRit; FIG. 5F), or rituximab alone (FIG.
5G).
DETAILED DESCRIPTION
[0048] After reading this description it will become apparent to
one skilled in the art how to implement the invention in various
alternative embodiments and alternative applications. However, all
the various embodiments of the present invention will not be
described herein. It will be understood that the embodiments
presented here are presented by way of an example only, and not
limitation. As such, this detailed description of various
alternative embodiments should not be construed to limit the scope
or breadth of the present invention as set forth below.
[0049] Before the present invention is disclosed and described, it
is to be understood that the aspects described below are not
limited to specific compositions, methods of preparing such
compositions, or uses thereof as such may, of course, vary. It is
also to be understood that the terminology used herein is for the
purpose of describing particular aspects only and is not intended
to be limiting.
[0050] The detailed description of the invention is divided into
various sections only for the reader's convenience and disclosure
found in any section may be combined with that in another section.
Titles or subtitles may be used in the specification for the
convenience of a reader, which are not intended to influence the
scope of the present invention.
Definitions
[0051] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. In this
specification and in the claims that follow, reference will be made
to a number of terms that shall be defined to have the following
meanings:
[0052] The terminology used herein is for the purpose of describing
particular embodiments only and is not intended to be limiting of
the invention. As used herein, the singular forms "a", "an" and
"the" are intended to include the plural forms as well, unless the
context clearly indicates otherwise.
[0053] "Optional" or "optionally" means that the subsequently
described event or circumstance can or cannot occur, and that the
description includes instances where the event or circumstance
occurs and instances where it does not.
[0054] The term "about" when used before a numerical designation,
e.g., temperature, time, amount, concentration, and such other,
including a range, indicates approximations which may vary by (+)
or (-) 10%, 5%,1%, or any subrange or subvalue there between.
Preferably, the term "about" when used with regard to a dose amount
means that the dose may vary by +/-10%. For example, "about 400 to
about 800 binding agents" indicates that an outside surface of a
nanoparticles contain an amount of binding agent between 360 and
880 particles.
[0055] "Comprising" or "comprises" is intended to mean that the
compositions and methods include the recited elements, but not
excluding others. "Consisting essentially of" when used to define
compositions and methods., shall mean excluding other elements of
any essential significance, to the combination for the stated
purpose. Thus, a composition consisting essentially of the elements
as defined herein would not exclude other materials or steps that
do not materially affect the basic and novel characteristic(s) of
the claimed invention. "Consisting of" shall mean excluding more
than trace elements of other ingredients and substantial method
steps. Embodiments defined by each of these transition terms are
within the scope of this invention.
[0056] The term "nanoparticle" as used herein refers to particles
having at least one dimension which is less than 5 microns, In
preferred embodiments, such as for intravenous administration, the
nanoparticle is less than 1 micron. For direct administration, the
nanoparticle is larger. Even larger particles are expressly
contemplated by the invention.
[0057] In a population of particles, the sizes of individual
particles are distributed about a mean. Particle sizes for the
population can therefore be represented by an average, and also by
percentiles. D50 is the particle size below which 50% of the
particles fall. 10% of particles are smaller than the D10 value and
90% of particles are smaller than D90. Where unclear, the "average"
size is equivalent to D50. So, for example., AB160 and AR.160 refer
to nanoparticles having an average size of 160 nanometers.
[0058] The term "nanoparticle" may also encompass discrete
multimers of smaller unit nanoparticles. For 160 nm nanoparticles,
multimers would therefore be approximately 320 nm, 480 nm, 640 nm,
800 nm, 960 nm, 1120 nm, and so on.
[0059] The term "carrier protein" as used herein refers to proteins
that function to transport binding agents and/or therapeutic
agents. The binding agents of the present disclosure can reversibly
bind to the carrier proteins. Examples of carrier proteins are
discussed in more detail below.
[0060] The term "core" as used herein refers to a central or inner
portion of the nanoparticle which may be comprised of a carrier
protein, a carrier protein and a therapeutic agent, or other agents
or combination of agents. In some embodiments, an albumin-binding
motif of the binding agent may be associated with the core.
[0061] The term "therapeutic agent" as used herein means an agent
which is therapeutically useful, e.g., an agent for the treatment,
remission or attenuation of a disease state, physiological
condition, symptoms, or etiological factors, or for the evaluation
or diagnosis thereof. A therapeutic agent may be a chemotherapeutic
agent, for example, mitotic inhibitors, topoisomerase inhibitors,
steroids, anti-tumor antibiotics, antimetabolites, alkylating
agents, enzymes, proteasome inhibitors, or any combination
thereof.
[0062] As used herein, the term, "binding agent", "binding agent
specific for", or "binding agent that specifically binds" refers to
an agent that binds to a target antigen and does not significantly
bind to unrelated compounds. Examples of binding agents that can be
effectively employed in the disclosed methods include, but are not
limited to, lectins, proteins, and antibodies, such as monoclonal
antibodies, e,g. humanized monoclonal antibodies, chimeric
antibodies, or polyclonal antibodies, or antigen-binding fragments
thereof, as well as aptamers fusion proteins, and aptamers having
or fused to an albumin-binding motif. In an embodiment the binding
agent is an exogenous antibody. An exogenous antibody is an
antibody not naturally produced in a mammal, e.g. in a human, by
the mammalian immune system.
[0063] The term "antibody" or "antibodies" as used herein refers to
immunoglobulin molecules and immunologically active portions of
immunoglobulin molecules (i.e., molecules that contain an antigen
binding site that immuno-specifically bind an antigen). The term
also refers to antibodies comprised of two immunoglobulin heavy
chains and two immunoglobulin light chains as well as a variety of
forms including full length antibodies and portions thereof;
including, for example, an immunoglobulin molecule, a monoclonal
antibody, a chimeric antibody, a CDR-grafted antibody, a humanized
antibody, a Fab, a Fab', a F(ab')2, Fv, a disulfide linked Fv, a
scFv, a single domain antibody (dAb), a diabody, a multispecific
antibody, a dual specific antibody, an anti-idiotypic antibody, a
bispecific antibody, a functionally active epitope-binding fragment
thereof, bifunctional hybrid antibodies (e.g., Lanzavecchia et al.,
Eur. J Immunol. 17, 105 (1987)) and single chains (e.g., Huston et
al., Proc. Nalt. Acad. Sci. US.A., 85, 5879-5883 (1988) and Bird et
al., Science 242, 423-426 (1988), which are incorporated herein by
reference). (See, generally, Hood et al., Immunology, Benjamin,
N.Y., 2ND ed. (1984); Harlow and Lane, Antibodies. A Laboratory
Manual, Cold Spring Harbor Laboratory (1988); Hunkapiller and Hood,
Nature, 323, 15-16 (1986), which are incorporated herein by
reference). The antibody may be of any type (e.g., IgG, IgA, IgE or
IgD). Preferably, the antibody is IgG. An antibody may be non-human
(e.g., from mouse, goat, or any other animal), fully human,
humanized, or chimeric. Antibody or antibodies include any
biosimilar(s) of the antibodies disclosed herein. Biosimilars, as
used herein, refers to a biopharmaceutical which is deemed to be
comparable in quality, safety, and efficacy to a reference product
marketed by an innovator company (Section 351(i) of the Public
Health Service Act (42 U.S.C. 262(i)).
[0064] The term "dissociation constant," also referred to as
"K.sub.d," refers to a quantity expressing the extent to which a
particular substance separates into individual components (e.g.,
the protein carrier, antibody, and a therapeutic agent).
[0065] The terms "lyophilized," "lyophilization" and the like as
used herein refer to a process by which the material (e.g.,
nanoparticles) to be dried is first frozen and then the ice or
frozen solvent is removed by sublimation in a vacuum environment.
An excipient is optionally included in pre-lyophilized formulations
to enhance stability of the lyophilized product upon storage. In
some embodiments, the nanoparticles can be formed from lyophilized
components (carrier protein, paclitaxel, and optionally antibody
and/or a therapeutic agent) prior to use as a therapeutic. In other
embodiments, the carrier protein and paclitaxel, and optionally a
binding agent, e.g., antibody, and/or a therapeutic agent are first
combined to facilitate the formation of stable nanoparticles and
then lyophilized. The lyophilized sample may further contain
additional excipients.
[0066] The term "bulking agents" comprise agents that provide the
structure of the freeze-dried product. Common examples used for
bulking agents include mannitol, glycine, lactose and sucrose. In
addition to providing a pharmaceutically elegant cake, bulking
agents may also impart useful qualities in regard to modifying the
collapse temperature, providing freeze-thaw protection, and
enhancing the protein stability over long-term storage. These
agents can also serve as tonicity modifiers. In some embodiments,
the lyophilized compositions described herein comprise bulking
agents. In some embodiments, the lyophilized compositions described
herein do not comprise bulking agents.
[0067] The term "buffer" encompasses those agents which maintain
the solution in an acceptable range prior to lyophilization and may
include succinate (sodium or potassium), histidine, phosphate
(sodium or potassium), Tris(tris(hydroxymethyl)aminomethane),
diethanolamine, citrate (sodium) and the like. The buffer of this
invention may have a pH in an the range from about 5.5 to about
6.5; and preferably has a pH of about 6.0. Examples of buffers that
will control the pH in this range include succinate (such as sodium
succinate), gluconate, histidine, citrate and other organic acid
buffers.
[0068] The term "cryoprotectants" generally includes agents which
provide stability to the protein against freezing-induced stresses,
presumably by being preferentially excluded from the protein
surface. They may also offer protection during primary and
secondary drying, and long-term product storage. Examples are
polymers such as dextran and polyethylene glycol; sugars such as
sucrose, glucose, trehalose, and lactose; surfactants such as
polysorbates; and amino acids such as glycine, arginine, and
serine.
[0069] The term "lyoprotectant" includes agents that provide
stability to the protein during the drying or "dehydration" process
(primary and secondary drying cycles), presumably by providing an
amorphous glassy matrix and by binding with the protein through
hydrogen bonding, replacing the water molecules that are removed
during the drying process. This helps to maintain the protein
conformation, minimize protein degradation during the
lyophilization cycle and improve the long-term products. Examples
include polyols or sugars such as sucrose and trehalose.
[0070] The term "pharmaceutical formulation" refers to preparations
which are in such form as to permit the active ingredients to be
effective, and which contains no additional components that are
toxic to the subjects to which the formulation would be
administered.
[0071] "Pharmaceutically acceptable" excipients (vehicles,
additives) are those which can reasonably be administered to a
subject mammal to provide an effective dose of the active
ingredient employed.
[0072] "Reconstitution time" is the time that is required to
rehydrate a lyophilized formulation into a solution.
[0073] A "stable" formulation is one in which the protein therein
essentially retains its physical stability and/or chemical
stability and/or biological activity upon storage. For example,
various analytical techniques for measuring protein stability are
available in the art and are reviewed in Peptide and Protein Drug
Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York,
N.Y., Pubs. (1991) and ,tones, A. Adv. Drug Delivery Rev. 10:29-90
(1993). Stability can be measured at a selected temperature for a
selected time period.
[0074] The term "epitope" as used herein refers to the portion of
an antigen which is recognized by a binding agent, e.g., an
antibody. Epitopes include, but are not limited to, a short amino
acid sequence or peptide (optionally glycosylated or otherwise
modified) enabling a specific interaction with a protein (e.g., an
antibody) or ligand. For example, an epitope may be a part of a
molecule to which the antigen-binding site of a binding agent
attaches.
[0075] The term "treating" or "treatment" covers the treatment of a
disease or disorder (e.g., cancer), in a subject, such as a human,
and includes: (i) inhibiting a disease or disorder, i.e., arresting
its development; (ii) relieving a disease or disorder, i.e.,
causing regression of the disease or disorder; (iii) slowing
progression of the disease or disorder; and/or (iv) inhibiting,
relieving, or slowing progression of one or more symptoms of the
disease or disorder. In some embodiments "treating" or "treatment"
refers to the killing of cancer cells.
[0076] The term "kill" or "killing" with respect to a cancer
treatment includes any type of manipulation that will directly or
indirectly lead to the death of that cancer cell or at least of
portion of a population of cancer cells.
[0077] The term "aptamer" refers to a nucleic acid molecule that is
capable of binding to a target molecule, such as a polypeptide. For
example, an aptamer of the invention can specifically bind to e.g.,
CD20, CD38, CD52, PD-1, PD-L1, PD-L2, Ly6E, HER2, HER3/EGFR DAF,
ERBB -3 receptor, CSF-1R, STEAP1, CD3, CEA, CD40, OX40, Ang2-VEGF
and VEGF. The generation of antibodies with a particular binding
specificity and the therapeutic use of aptamers are well
established in the art. See, e.g., U.S. Pat. Nos. 5,475,096,
5,270,163, 5,582,981, 5,840,867, 6,011,020, 6,051,698, 6,147,204,
6,180,348 and 6,699,843, and the therapeutic efficacy of
Macugen.RTM. (Eyetech, New York) for treating age-related macular
degeneration, each of which is incorporated herein by reference in
its entirety.
[0078] The term "oligomer" or "oligomeric" or "oligomerized" as
used herein refers to oligomers composed of two or more
monomers.
[0079] Fusion proteins are bioengineered polypeptides that join a
protein or fragment thereof (e.g., the crystallizable fragment (Fc)
domain of an antibody) with another biologically active agent
(e.g., a protein domain, peptide, or nucleic acid or peptide
aptamer) to generate a molecule with desired structure-function
properties and significant therapeutic potential. The gamma
immunoglobulin (IgG) isotype is often used as the basis for
generating Fc-fusion proteins because of favorable characteristics
such as recruitment of effector function and increased plasma
half-life. Given the range of aptamers, both peptide and nucleic
acids, that can be used as fusion partners, fusion proteins ave
numerous biological and pharmaceutical applications.
[0080] The term "non-toxic nanoparticles" (NTPs) or "NTP complexes"
refers to nanoparticles comprising a carrier protein (e.g.,
albumin) and a paclitaxel derivative that is less toxic than
paclitaxel. Optionally, the NTPs or NTP complexes comprise binding
agents (e.g., antibodies) and/or therapeutic agents (e.g.,
chemotherapeutic agents).
[0081] The phrase "less toxic than paclitaxel" refers to paclitaxel
derivatives that exhibit reduced toxicity to (e.g., reduced killing
of) cells, including cancer cells and normal cells, as compared to
paclitaxel. For example, the Meerwein Product of Paclitaxel
(20-Acetoxy-4-deactyl-5-epi-20, O-secotaxol) has significantly
reduced toxicity, likely due to the breaking of the C-4,C-5 oxetane
ring of paclitaxel.
[0082] The term "reduced toxicity nanoparticles" (RTPs) or "RTP
complexes" refers to nanoparticles comprising a carrier protein
(e.g., albumin) and a reduced amount of paclitaxel compared to
ABRAXANE.RTM. (.g., an albumin:paclitaxel ratio of about 9:1).
Optionally, the RTPs or RIP complexes comprise binding agents
(e.g., antibodies) and/or therapeutic agents (e.g.,
chemotherapeutic agents).
[0083] Additionally, some terms used in this specification are more
specifically defined below.
Overview
[0084] ABRAXANE.RTM. for Injectable Suspension (paclitaxel
protein-bound particles for injectable suspension) is an
albumin-bound form of paclitaxel with a mean particle size of
approximately 130 nanometers. Paclitaxel can exist in the particles
in a non-crystalline, amorphous state. ABRAXANE.RTM. can be
supplied as, for example, a lyophilized powder for reconstitution
with 20 mL of 0.9% Sodium Chloride Injection, USP prior to
intravenous infusion. A single-use vial contains 100 mg of
paclitaxel and approximately 900 mg of human albumin. Each
milliliter (mL) of reconstituted suspension contains 5 mg
paclitaxel.
[0085] ABRAXANE.RTM. nanoparticles are stabilized albumin-bound
paclitaxel. The association of the paclitaxel and albumin can be
via, for example, hydrophobic interactions. In some examples, the
nanoparticle are stable at an average size of 130 nm. It is also
known that other hydrophobic drugs such as, for example, docetaxel,
rapamycin, can similarly form stabilized nanoparticles by binding
to albumin.
[0086] For conventional ADCs to be effective, it is critical that
the linker be stable enough not to dissociate in the systemic
circulation but allow for sufficient drug release at the tumor
site. Alley, S. C., et al. (2008) Bioconjug Chem 19:759-765. This
has proven to be a major hurdle in developing effective drug
conjugate (Julien, D.C., et al. (2011) MAbs 3:467-478; Alley, S.
C., et al. (2008) Bioconjug Chem 19:759-765); therefore, an
attractive feature of the nano-immune conjugate is that a
biochemical linker is not required.
[0087] Another shortcoming of current ADCs is that higher drug
penetration into the tumor has not been substantively proven in
human tumors. Early testing of ADCs in mouse models suggested that
tumor targeting with antibodies would result in a higher
concentration of the active agent in the tumor (Deguchi, T. et al.
(1986) Cancer Res 46: 3751-3755); however, this has not correlated
in the treatment of human disease, likely because human tumors are
much more heterogeneous in permeability than mouse tumors, Jain, R.
K. et al. (2010) Nat Rev Clin Oncol 7:653-664. Also, the size of
the nanoparticle is critical for extravasation from the vasculature
into the tumor. In a mouse study using a human colon adenocarcinoma
xenotransplant model, the vascular pores were permeable to
liposomes up to 400 nm. Yuan, F., et al. (1995) Cancer Res 55:
3752-3756. Another study of tumor pore size and permeability
demonstrated that both characteristics were dependent on tumor
location and growth status, with regressing tumors and cranial
tumors permeable to particles less than 200 nm. Hobbs, S. K., et
al., (1998) Proc Natl Acad. Sci USA 95:4607-4612. The nano-immune
conjugate described herein overcomes this issue by the fact that
the large complex, which is less than 200 nm intact is partially
dissociated in systemic circulation into smaller functional units
that are easily able to permeate tumor tissue. Furthermore, once
the conjugate arrives to the tumor site, the smaller toxic payload
can be released and only the toxic portion needs to be taken up by
tumor cells, not the entire conjugate.
[0088] The advent of antibody- (i.e. AVASTIN.RTM.) coated albumin
nanoparticles containing a therapeutic agent (i.e., ABRAXANE.RTM.)
has led to a new paradigm of directional delivery of two or more
therapeutic agents to a predetermined site in vivo. See PCT Patent
Publication Nos. WO 2012/154861 and WO 2014/055415, each of which
is incorporated herein by reference in its entirety.
[0089] However, it is contemplated that some cancers will not be
treated by the paclitaxel in the ABRAXANE.RTM.-antibody complexes,
and/or that other therapeutic agents (e.g., anti-cancer
chemotherapeutic agents) will be more effective in treating certain
cancers. Herein are disclosed reduced toxicity nanoparticles (RIP)
comprising a carrier protein (e.g., albumin) and a reduced amount
of paclitaxel (e.g., compared to ABRAXANE.RTM.), or non-toxic
nanoparticles (NTP) comprising a carrier protein (e.g., albumin)
and a paclitaxel derivative that is less toxic than paclitaxel. The
nanoparticles may further include a therapeutic agent and/or
binding agents (e.g., antibodies).
[0090] Nanoparticles and Nanoparticle Compositions
[0091] As will be apparent to the skilled artisan upon reading this
disclosure, the present disclosure relates to nanoparticles and
compositions of nanoparticles containing a carrier protein (e.g,
albumin) and a paclitaxel derivative (e.g., a derivative that is
less toxic than paclitaxel) or a reduced amount of paclitaxel
(e.g., compared to a therapeutic amount, and/or compared to an
amount in ABRAXANE.RTM.). In some embodiments, the nanoparticles
also contain binding agents and/or a therapeutic agent. In some
embodiments, the nanoparticles or nanoparticle composition is
lyophilized.
[0092] The present invention is further predicated, in part, on the
formation of nanoparticles comprising a carrier protein and
paclitaxel in a lower amount than is present in ABRAXANE.RTM. or a
paclitaxel derivative that is less toxic (e.g., to cells) than
paclitaxel, optionally with a binding agent and/or a therapeutic
agent. Without being hound by theory, it is believed that such
nanoparticles provide targeted therapy to a tumor while minimizing
toxicity to the patient, and can be used to treat tumors that
traditionally are not susceptible to paclitaxel and/or that respond
better to a different therapeutic agent.
[0093] In some embodiments, the cation protein can be albumin,
gelatin, elastin (including topoelastin) or elastin-derived
polypeptides (e.g., .alpha.-elastin and elastin-like polypeptides
(ELPs)), gliadin, legumin, zein, soy protein (e.g., soy protein
isolate (SPI)), milk protein (e.g., .beta.-lactoglobulin (BLG) and
casein), or whey protein (e.g, whey protein concentrates (WPC) and
whey protein isolates (WPI)). In preferred embodiments, the carrier
protein is albumin. In preferred embodiments, the albumin is egg
white (ovalbumin), bovine serum albumin (BSA), or the like. In even
more preferred embodiments, the carrier protein is human serum
albumin (HSA). In some embodiments, the carrier protein is a
recombinant protein (e,g., recombinant HSA). In some embodiments,
the carrier protein is a generally regarded as safe (GRAS)
excipient approved by the United States Food and Drug
Administration (FDA).
[0094] In some embodiments, the binding agents are antibodies
selected from adotrastuzumab emtansine, alemtuzumab, atezolizumab,
beyacizumab, cetuximab, denosumab, dinutuximab, nivolumab,
obinutuzumab, ofatumumab, panitumuinab, petnbrolizumab, pertuzumab,
rituximab, avelumab tar durvalumab, pidilizumab, BMS 936559, and
trastuzumab. In some embodiments, the antibodies are selected from
the antibodies listed in Table 1. In some embodiments, the
antibodies are a substantially single layer of antibodies on all or
part of the surface of the nanoparticle.
[0095] Table 1 depicts a list of non-limiting list of
antibodies.
TABLE-US-00001 TABLE 1 Antibodies Biologic Treatment(s)/Target(s)
Rituximab (Rituxan .RTM.) Non-Hodgkin lymphoma AlemtuzumaB (Campath
.RTM.) Chronic lymphocytic leukemia (CLL) Ipilimumab (Yervoy .RTM.)
Metastatic melanoma Bevacizumab (Avastin .RTM.) Colon cancer, lung
cancer, renal cancer, ovanan cancer, glioblastoma multiforme
Cetuximab (Erbitux .RTM.) Colorectal cancer, non-small cell lung
cancer, head and neck cancer, cervical cancer, glioblastoma,
ovarian epithelia, fallopian tube or primary peritoneal cancer,
renal cell cancer Panitumumab (Vectibix .RTM.) Colorectal cancer
Trastuzumab (Herceptin .RTM.) Breast cancer, Adenocarcinoma
.sup.90Y-ibritumomab Non-Hodgkin lymphoma tiuxetan (Zevalin .RTM.)
Brentuximab vedotin Hodgkin lymphoma, Anaplastic large cell
lymphoma (Adcetris .RTM.) Blinatumomab (Blincyto) Acute lymphocytic
leukemia (ALL) Pembrolizumab (Keytruda .RTM.) PD-1 (melanoma,
non-small cell lung cancer) Nivolumab (Opdivo .RTM.) PD-1
(melanoma, non-small cell lung cancer) Ofatumumab (Arzerra .RTM.)
Chronic lymphocytic leukemia (CLL) Pertuzumab (Perieta .RTM.)
Breast cancer Obinutuzumab (Gazyva .RTM.) Lymphoma, diffuse large
B-cell lymphoma (DLBCL), indolent NEIL (1st-line) Din u tux imab
(Unitux jn .RTM.) Neuroblastoma Denosurnab (Prolia .RTM.) Bone
metastases, multiple myeloma, giant cell tumor of bone RG6016 (LSD1
inhibitor) mAB Acute myelogenous leukemia (AML) Small molecule
according to BioCentury BCIQ RG7882 (antibody drug Pancreatic
cancer, ovarian cancer conjugate) Alternative Names: D-4064A; DMUC
4064A; RG7882 Lifastuzumab vedotin (antibody Platinum-resistant
ovarian cancer, NSCLC drug conjugate) Polatuzumab vedotin (antibody
DLBCL, NHL drug conjugate) RG7446 (anti-PD-L1 mAb) bladder cancer,
NSCLC, melanoma, breast, renal cell carcinoma, lymphoma
Atezolizumab (Tecentriq .RTM., Bladder cancer, metastatic NSCLC
anti-PD-L1) DLYE-5953A (anti-Ly6E mAB Refractory solid tumors
cytotoxic drug conjugate) Duligotuzumab Solid tumors with mutant
KRAS (anti-HER3/EGFR DAF mAb) RG7117 (ERBB-3 receptor Metastatic
breast cancer antagonist) RG7155 (CSF-1R antagonist) Solid tumors
RG-7450 (anti-STEAP1 antibody Prostate cancer drug conjugate)
RG7802 (CD3/CEA bispecific Solid tumors antibody) RG7813 (CEA
inhibitor) Solid tumors RG7841 (antibody drug Solid tumors
conjugate) RG7876 (CD40 antigen Solid tumors stimulant) RG7888
(anti-OX40 mAb) Solid tumors RG7221 (Ang2-VEGF mAb) Metastatic
colorectal cancer RG7686 (glypican-3 mAb) Hepatocellular carcinoma
Perjeta .RTM. pertuzumab HER3-positive breast cancer, gastric
cancer Avelumab (anti-PD-L1 mAb) Solid tumor, gastric cancer,
Merkel cell carcinoma, non-small cell lung cancer Durvalumab
(anti-PD-L1 mAb) NSCLC, head and neck, bladder, gastric,
pancreatic, HCC and blood cancers Pidilizumab/CT-011 Lymphoma,
myeloma (anti-PD-1 mAb) BMS 936559/MDX-1105 melanoma, non-small
cell lung cancer (anti-PD-L1 mAb)
[0096] In some embodiments, the at least one therapeutic agent is
selected from ahiraterone, bendamustine, bortezomib, carboplatin,
cabazitaxel, cisplatin, chlorambucil, dasatinib, docetaxel,
doxorubicin, epirubicin, erlotinib, etoposide, everolimus,
gefitinib, idarubicin, imatinib, hydroxyurea, imatinib, lapatinib,
leuprorelin, melphalan, methotrexate, mitoxantrone, nedaplatin,
nilotinib, oxaliplatin, pazopanib, pemetrexed, picoplatin,
romidepsin, satraplatin, sorafenib, vernurafenib, sunitinib,
teniposide, triplatin, vinblastine, vinorelbine, vincristine, and
cyclophosphamide.
[0097] Table 2 depicts a list of non-limiting list of cancer
therapeutic agents. In one embodiment, the therapeutic agent is
selected from the agents recited in Table 2.
TABLE-US-00002 TABLE 2 Cancer therapeutic agents Cancer Drugs Drug
Target(s) Abitrexate (Methotrexate) Acute lymphoblastic leukemia;
breast cancer; gestational trophoblastic disease, head and neck
cancer; lung cancer; mycosis fungoides non-Hodgkin lymphoma;
osteosarcoma Ado-Trastuzumab Emtansine Breast cancer Adriamycin
(Doxorubicin Hydrochloride) Acute lymphoblastic leukemia; acute
myeloid leukemia; breast cancer, gastric (stomach) cancer; Hodgkin
lymphoma neuroblastoma; non-Hodgkin lymphoma; ovarian cancer; small
cell lung cancer; soft tissue and bone sarcomas; thyroid cancer;
transitional cell bladder cancer; Wilms tumor Adrucil, Efudex,
Fluoroplex (Fluoromacil) Basal cell carcinoma; breast cancer;
colorectal cancer; gastric (stomach) adenocarcinoma; pancreatic
cancer; squamous cell carcinoma of the head and neck Afinitor
(Everolimus) Breast cancer, pancreatic cancer; renal cell
carcinoma; subependymal giant cell astrocytoma Alimta (Pemetrexed
Disodium) Malignant pleural mesothelioma; non-small cell lung
cancer Ambochlorin, Leukeran, or Linfolizin Chronic lymphocytic
leukemia; Hodgkin (Chlorambucil) lymphoma; non-Hodgkin lymphoma
Aredia (Pamidronate Disodium) Breast cancer; multiple myclotna.
Arimidex (Anastrozole) Breast cancer Aromasin (Exemestane) Advanced
breast cancer; early-stage breast cancer and estrogen receptor
positive Arranon (Nelarabine) T-cell acute lymphoblastic leukemia;
T-cell lymphoblastic lymphoma BEACOPP Hodgkin lymphoma Becenum,
BiCNU (Carmustine) Brain tumors; Hodgkin lymphoma; multiple
myeloma; non-Hodgkin lymphoma Beleodaq (Belinostat) Peripheral
T-cell lymphoma BEP Ovarian germ cell tumors; testicular germ cell
tumors Bleomycin Hodgkin lymphoma; non-Hodgkin lymphoma; penile
cancer; squamous cell carcinoma of the cervix; squamous cell
carcinoma of the head and neck; squamous cell carcinoma of the
vulva; testicular cancer Bosulif (Bosutinib) Chronic myelogenous
leukemia Busulfex or Myleran (Busulfan) Chronic myelogenous
leukemia CAF Breast cancer Camptosar (Irinotecan Hydrochloride)
Colorectal cancer CAPOX Colorectal cancer Casodex (Bicalutamide)
Prostate cancer CccNU (Lomustine) Brain tumors; Hodgkin lymphoma
Ceritinib Non-small cell lung cancer CHOP Non-Hodgkin lymphoma
Clofarex (Clofarabine) Acute lymphoblastic leukemia CMF Breast
cancer Cometriq (Cabozantinib-S-Malate) Medullary thyroid cancer
COPP Hodgkin lymphoma; non-Hodgkin lymphoma COPP-ABV Hodgkin
lymphoma Cosmegen (Dactinomycin) Ewing sarcoma; gestational
trophoblastic disease; rhabdomyosarcoma; solid tumors; testicular
cancer; Wilms tumor CVP Non-Hodgkin lymphoma; chronic lymphocytic
leukemia Cyfos (Ifosfamide) Testicular germ cell tumors Cytoxan or
Neosar (Cyclophosphamide) Acute lymphoblastic leukemia; acute
myeloid leukemia; breast cancer; chronic lymphocytic leukemia;
chronic myelogenous leukemia; Hodgkin lymphoma; multiple myeloma;
mycosis fungoides; neuroblastoma; non- Hodgkin lymphoma; ovarian
cancer; retinoblastoma Dacarbazine Hodgkin lymphoma; melanoma
Dacogen (Decitabine) Myelodysplastic syndromes Degarelix Prostate
cancer Denileukin Diftitox Cutaneous T-cell lymphoma Denosumab
Giant cell tumor of the bone; breast cancer, prostate cancer
DepoCyt ar DepoFoam (Liposomal Cytarabine) Lymphomatous meningitis
DTIC-Dome (Dacarbazine) Hodgkin lymphoma; melanoma Ellence
(Epirubicin Hydrochloride) Breast cancer Eloxatin (Oxaliplatin)
Colorectal cancer; stage III colon cancer Emend (Aprepitant) Nausea
and vomiting caused by chemotherapy and nausea and vomiting after
surgery EPOCH Non-Hodgkin lymphoma Erbitux (Cetuximab) Colorectal
cancer; squamous cell carcinoma of the head and neck Eribulin
Mesylate Breast cancer Erivedge (Vismodegib) Basal cell carcinoma
Erlotinib Hydrochloride Non-small cell lung cancer; pancreatic
cancer Erwinaze (Asparaginase Acute lymphoblastic leukemia Erwinia
chrysanthemi) Etopophos (Etoposide Phosphate) Small cell lung
cancer; testicular cancer Evacet or LipoDox or Doxil AIDS-related
Kaposi sarcoma; multiple (Doxorubicin Hydrochloride Liposome)
myeloma; ovarian cancer Evista or Keoxifene (Raloxifene
Hydrochloride) Breast cancer Fareston (Toremifene) Breast cancer
Farydak (Panobinostat) Multiple myeloma Faslodex (Fulvestrant)
Breast cancer FEC Breast cancer Femara (Letrozole) Breast cancer
Filgrastim Neutropenia Fludara (Fludambine Phosphate) Chronic
lymphocytic leukemia FOLFIRI Colorectal cancer FOLFIRI-BEVACIZUMAB
Colorectal cancer FOLFIRI-CETUXIMAB Colorectal cancer FOLFIRINOX
Pancreatic cancer FOLFOX Colorectal cancer Folotyn (Pralatrexate)
Peripheral T-cell lymphoma FU-LV Colorectal cancer; esophageal
cancer; gastric cancer GEMCITABINE-CISPLATIN Biliary tract cancer;
bladder cancer; cervical cancer; malignant mesothelioma; non-small
cell lung cancer; ovarian cancer; pancreatic cancer
GEMCITABINE-OXALIPLATIN Pancreatic cancer Gemzar (Gemcitabine
Hydrochloride) Breast cancer; non-small cell lung cancer; ovarian
cancer; pancreatic cancer Gilotrif (Afatinib Dimaleate) Non-small
cell lung cancer Gleevcc (Imatinib Mesylate) Acute lymphoblastic
leukemia; chronic eosinophilic leukemia or hypereosinophilic
syndrome; chronic myelogenous leukemia; dermatofibrosarcoma
protuberans; gastrointestinal stromal tumor;
myelodysplastic/myeloproliferative neoplasms; systemic
mastocytosis. Gliadel (Carmustine Implant) Glioblastoma multiforme;
malignant glioma Halaven (Eribulin Mesylate) Breast cancer Hycamtin
(Topotecan Hydrochloride) Cervical cancer; ovarian cancer; small
cell lung cancer Hyper-CVAD Acute lymphoblastic leukemia;
non-Hodgkin lymphoma Ibrance (Palbociclib) Breast cancer ICE
Hodgkin lymphoma: non-Hodgkin lymphoma Iclusig (Ponatinib
Hydrochloride) Acute lymphoblastic leukemia, Chronic myelogenous
leukemia Idamycin (Idarubicin Hydrochloride) Acute myeloid leukemia
Imbruvica (Ibruitinib) Chronic lymphocytic, leukemia; mantle cell
lymphoma; Waldenstr6m macroglobulinemia Inlyta (Axitinib) Renal
cell carcinoma Iressa (Gefitinib) Non-small cell lung cancer
Istodax (Romidepsin) Cutaneous T-cell lymphoma Ixempra
(Ixabepilone) Breast cancer Jevtana (Cabazitaxel) Prostate cancer
Kyprolis (Carfilzomib) Multiple myeloma Lenvima (Lenvatinib
Mesylate) Thyroid cancer Leuprolide Acetate Prostate cancer Lupron
(Leuprolide Acetate) Prostate cancer Lynparza (Olaparib) Ovarian
cancer Marqibo (Vincristine Sulfate Liposome) Acute lymphoblastic
leukemia Matulane (Procarbazine Hydrochloride) Hodgkin lymphoma
Megace (Megestrol Acetate) Breast cancer; endometrial cancer
Mekinist (Trametinib) Melanoma Mesnex (Mesna) Hemorrhagic cystitis
Mitoxantrone Hydrochloride Acute myeloid leukemia; prostate cancer
Mitozytrex (Mitomycin C) Gastric (stomach) and pancreatic
adenocarcinoma MOPP Hodgkin lymphoma Mozobil (Plerixafor) Multiple
myeloma; non-Hodgkin lymphoma Mustargen Bronchogenic carcinoma,
chronic lymphocytic (Mechlorethamine Hydrochloride) leukemia;
chronic myelogenous leukemia; Hodgkin lymphoma; malignant pleural
effusion, malignant pericardial effusion, and malignant peritoneal
effusion; mycosis fungoides; non-Hodgkin lymphoma Mylotarg
(Gemtuzumab Ozogamicin) Acute myeloid leukemia Navelbine
(Vinorelbine Tartrate) Non-small cell lung cancer Nexavar
(Sorafenib Tosylate) Hepatocellular carcinoma; Renal cell
carcinoma; Thyroid cancer Nilotinib Chronic myelogenous leukemia
Nolvadex (Tamoxifen Citrate) Breast cancer Odomzo (Sonidegib) Basal
cell carcinoma OEPA Hodgkin lymphoma OFF Pancreatic cancer Oncaspar
(Pegaspargase) Acute lymphoblastic leukemia OPPA Hodgkin lymphoma
Paclitaxel AIDS-related Kaposi sarcoma; Breast cancer; Non-small
cell lung cancer; Ovarian cancer PAD Multiple myeloma Paraplat
(Carboplatin) Non-small cell lung cancer; Ovarian cancer Paraplatin
(Carboplatin) Non-small cell lung cancer; Ovarian cancer Platinol
(Cisplatin) Bladder cancer; Cervical cancer; Malignant
mesothelioma; Non-small cell lung cancer; Ovarian cancer; Squamous
cell carcinoma of the head and neck; Testicular cancer Pomalyst
(Pomalidomide) Multiple myeloma Pontinib Hydrochloride Acute
lymphoblastic leukemia; Chronic myelogenous leukemia Prednisone
Acute lymphoblastic leukemia; Chronic lymphocytic leukemia; Hodgkin
lymphoma; Multiple myeloma; Non-Hodgkin lymphoma; Prostate cancer;
Thymoma and thymic carcmoma Provenge (Sipuleucel-T) Prostate cancer
Purinethol (Mercaptopurine) Acute lymphoblastic leukemia Radium 223
Dichloride Prostate cancer R-CHOP Non-Hodgkin lymphoma R-CVP
Non-Hodgkin lymphoma R-EPOCH B-cell on-Hodgkin lymphoma Revlimid
(Lenalidomide) Mantle cell lymphoma; Multiple myeloma; Anemia
Rubidomycin (Daunorubicin Hydrochloride) Acute lymphoblastic
leukemia; Acute myeloid leukemia Sipuleucel-T Prostate cancer
Somatuline Depot (Lanreotide Acetate) Gastroenteropancreatic
neuroendocrine tumors Sprycel (Dasatinib) Acute lymphoblastic
leukemia; Chronic myelogenous leukemia. STANFORD V Hodgkin lymphoma
Stivarga (Regorafenib) Colorectal cancer; Gastrointestinal stromal
tumor Sutent (Sunitinib Malate) Gastronintestinal stromal tumor;
Pancreatic cancer; Renal cell carcinoma Synovir (Thalidomide)
Multiple myeloma Synribo (Omacetaxine Mepesuccinate) Chronic
myelogenous leukemia TAC Breast cancer Tafinlar (Dabrafenib)
Melanoma Tarabine PFS (Cytarabine) Acute lymphoblastic leukemia;
Acute myeloid leukemia; Chronic myelogenous leukemia; meningeal
leukemia Tarceva (Erlotinib Hydrochloride) Non-small cell lung
cancer; Pancreatic cancer Targretin (Bexarotene) Skin problems
caused by cutaneous T-cell lymphoma Tasigna (Niltinib) Chronic
myelogenous leukemia Taxol (Paclitaxel) AIDS-related Kaposi
sarcoma; Breast cancer; Non-small cell lung cancer; Ovarian cancer
Taxotere (Docetaxel) Breast cancer; Adenocarcinoma; Non-small cell
lung cancer; Prostate cancer; Squamous cell carcinoma of the head
and neck; adenocarcinoma of the stomach or gastroesophageal
junction; Temodar or Methazolastone Anaplastic astrocytoma;
(Temozolomide) Glioblastoma multiforme Thiotepa Bladder cancer;
Breast cancer; Malignant pleural effusion, malignant pericardial
effusion, and malignant peritoneal effusion; Ovarian cancer Toposar
or VePesid (Etoposide) Small cell lung cancer; Testicular cancer
Torisel (Temsirolimus) Renal cell carcinoma TPF Squamous cell
carcinoma of the head and neck; Gastric (stomach) cancer Treanda
(Bendamustine Hydrochloride) B-cell non-Hodgkin lymphoma; Chronic
lymphocytic leukemia
Trisenox (Arsenic Trioxide) Acute promyelocytic leukemia Tykerb
(Lapatinib Ditosylate) Breast cancer Vandetabib Medullary thyroid
cancer VAMP Hodgkin lymphoma VeIP Ovarian germ cell; Testicular
cancer Velcade (Bortezomib) Mulitple myeloma; Mantle cell lymphoma
Velsar or Velban (Vinblastine Sulfate) Breast cancer;
Choriocarcinoma; Hodgkin lymphoma; Kaposi sarcoma; Mycosis
fungoides; Non-Hodgkin lymphoina; Testicular cancer Viadur
Leuprolide Acetate) Prostate cancer Vidaza (Azacitidine)
Myelodysplastic syndromes Vincasar PFS (Vincristine Sulfate) Acute
leukemia; Hodgkin lymphoma; Neuroblastoma; Non-Hodgkin lymphoma;
Rhabdomyosarcoma; Wilms tumor VIP Testicular cancer Visbodegib
Basal cell carcinoma Votrient (Pazopanib Hydrochloride) Renal cell
carcinoma; Soft tissue sarcoma Wellcovorin (Leucovorin Calcium)
Colorectal cancer; Anemia Xalkori (Crizotinib) Non-small cell lung
cancer Xeloda (Capecitabine) Breast cancer; Colorectal cancer
XELIRI Colorectal cancer; Esophageal cancer; Gastric (stomach)
cancer XELOX Colorectal cancer Xofigo (Radium 223 Dichloride)
Prostate cancer Xtandi (Enzalutamide) Prostate cancer Zaltrap
(Ziv-Afibercept) Colorectal cancer Zelboraf (Vemurafenib) Melanoma
Zoladex (Goserelin Acetate) Breast cancer; Prostate cancer Zolinza
(Vorinostat) Cutaneous T-cell lymphoma Zometa (Zoledronic Acid)
Multiple myeloma Zydelig (Idelalisib) Chronic lymphocytic leukemia;
Non-Hodgkin lymphoma (Follicula B-cell non Hodgkin lymphoma and
Small lymphocytic lymphoma) Zykadia (Certinib) Non-small cell lung
cancer Zytiga (Abiraterone Acetate) Prostate cancer
[0098] It is to be understood that the therapeutic agent may be
located inside the nanoparticle, on the outside surface of the
nanoparticle, or both. The nanoparticle may contain more than one
therapeutic agent, for example, two therapeutic agents, three
therapeutic agents, four therapeutic agents, five therapeutic
agents, or more. Furthermore, a nanoparticle may contain the same
or different therapeutic agents inside and outside the
nanoparticle.
[0099] In one aspect, the nanoparticle comprises at least 100
binding agents non-covalently bound to the surface of the
nanoparticle. In one aspect, the nanoparticle comprises at least
200, 300, 400, 500, 600, 700 or 800 binding agents non-covalently
bound to the surface of the nanoparticle.
[0100] In one aspect, the nanoparticle comprises between about 100
and about 1000 binding agents non-covalently bound to the surface
of the nanoparticle. In one aspect, the nanoparticle comprises
between about 200 and about 1000, between about 300 and about 1000,
between about 400 and about 1000, between about 500 and about 1000,
between about 600 and about 1000, between about 200 and about 800,
between about 300 and about 800, or between about 400 and about 800
binding agents non-covalently bound to the surface of the
nanoparticle. Contemplated values include any value or subrange
within any of the recited ranges, including endpoints.
[0101] In one aspect, the average particle size in the nanoparticle
composition is less than about 1 .mu.m. In one aspect, the average
particle size in the nanoparticle composition is between about 50
nm and about 1 nm. In one aspect, the average particle size in the
nanoparticle composition is between about 60 nm and about 900 nm.
In one aspect, the average particle size in the nanoparticle
composition is between about 60 nm and about 800 nm. In one aspect,
the average particle size in the nanoparticle composition is
between about 60 nm and about 700 nm. In one aspect, the average
particle size in the nanoparticle composition is between about 60
nm and about 600 nm. In one aspect, the average particle size in
the nanoparticle composition is between about 60 nm and about 500
nm. In one aspect, the average particle size in the nanoparticle
composition is between about 60 nm and about 400 nm. In one aspect,
the average particle size in the nanoparticle composition is
between about 60 nm and about 300 nm. In one aspect, the average
particle size in the nanoparticle composition is between about 60
nm and about 200 nm. In one aspect, the average particle size in
the nanoparticle composition is between about 80 nm and about 900
nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 mn, 300 nm, or 200 nm. In
one aspect, the average particle size in the nanoparticle
composition is between about 100 nm and about 900 nm, 800 nm, 700
nm, 600 nm, 500 nm, 400 nm, 300 nm, or 200 nm. In one aspect, the
average particle size in the nanoparticle composition is between
about 120 nm and about 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400
nm, 300 nm, or 200 nm. Contemplated values include any value,
subrange, or range within any of the recited ranges, including
endpoints.
[0102] In one aspect, the nanoparticle composition is formulated
for intravenous injection. In order to avoid an ischemic event, the
nanoparticle composition formulated for intravenous injection
should comprise nanoparticles with an average particle size of less
than about 1 .mu.m.
[0103] In one aspect, the average particle size in the nanoparticle
composition is greater than about 1 .mu.m. In one aspect, the
average particle size in the nanoparticle composition is between
about 1 .mu.m and about 5 .mu.m, between about 1 .mu.m and about 4
.mu.m, between about 1 pm and about 3 .mu.m, between about 1 .mu.m
and about 2 nm, or between about 1 .mu.m and about 1.5 .mu.m.
Contemplated values include any value, subrange, or range within
any of the recited ranges, including endpoints.
[0104] In one aspect, the nanoparticle composition is formulated
for direct injection into a tumor. Direct injection includes
injection into or proximal to a tumor site, perfusion into a tumor,
and the like. When formulated for direct injection into a tumor,
the nanoparticle may comprise any average particle size. Without
being bound by theory, it is believed that larger particles (e.g.,
greater than 500 nm, greater than 1 .mu.m, and the like) are more
likely to be immobilized within the tumor, thereby providing a
beneficial effect. Larger particles can accumulate in the tumor or
specific organs. See, e.g., 20-60 micron glass particle that is
used to inject into the hepatic artery feeding a tumor of the liver
called "THERASPHERE.RTM." (in clinical use for liver cancer).
Therefore, for intravenous administration, particles under 1 .mu.m
are typically used. Particles over 1 .mu.m are, more typically,
administered directly into a tumor ("direct injection") or into an
artery feeding into the site of the tumor.
[0105] In one aspect, less than about 0.01% of the nanoparticles
within the composition have a particle size greater than 200 nm,
greater than 300 nm, greater than 400 nm, greater than 500 nm,
greater than 600 nm, greater than 700 nm, or greater than 800 nm.
In one aspect, less than about 0.001% of the nanoparticles within
the composition have a particle size greater than 200 nm, greater
than 300 nm, greater than 400 nm, greater than 500 nm, greater than
600 nm, greater than 700 nm, or greater than 800 nm. In a preferred
embodiment, less than about 0.01% of the nanoparticles within the
composition have a particle size greater than 800 nm. In a more
preferred embodiment, less than about 0.001% of the nanoparticles
within the composition have a particle size greater than 800
nm.
[0106] In a preferred aspect, the sizes and size ranges recited
herein relate to particle sizes of the reconstituted lyophilized
nanoparticle composition. That is, after the lyophilized
nanoparticles are resuspended in an aqueous solution (e.g., water,
other pharmaceutically acceptable excipient, buffer, etc.), the
particle size or average particle size is within the range recited
herein.
[0107] In one aspect, at least about 50%, 60%, 70%, 80%, 90%, 95%,
96%. 97%., 98%, 99%, 99.5%, or 99.9% of the, nanoparticles are
present in the reconstituted composition as single nanoparticles.
That is, fewer than about 50%, 40%, 30%, etc. of the nanoparticles
are dimerized or multimerized (otigomerized).
[0108] In some embodiments, the nanoparticles in the composition
have less than 20% by number dimerization, less than 10% by number
dimerization and preferably less than 5% dimerization.
[0109] In some embodiments, the size of the nanoparticle can be
controlled by the adjusting the amount (e.g., ratio) of carrier
protein to paclitaxel or paclitaxel derivative, and/or ratio of
carrier protein-paclitaxel (or paclitaxel derivative) nanoparticles
to binding agent. The size of the nanoparticles, and the size
distribution, is also important. The nanoparticles of the invention
may behave differently according to their size. At large sizes, an
agglomeration may block blood vessels. Therefore, agglomeration of
nanoparticles can affect the performance and safety of the
composition. On the other hand, larger particles may be more
therapeutic under certain conditions (e.g., when not administered
intravenously).
[0110] Further challenges are imposed because the nanoparticles are
used in therapy.
[0111] While rearrangement of the components in the nanoparticle
may be mitigated through covalent bonds between the components,
such covalent bonds pose challenges for the therapeutic use of
nanoparticles in cancer treatment. The binding agent, carrier
protein, and therapeutic agent typically act at different locations
in a tumor and through different mechanisms. Non-covalent bonds
permit the components of the nanoparticle to dissociate at the
tumor, Thus, while a covalent bond may be advantageous for
lyophilization, it may he disadvantageous for therapeutic use.
[0112] The size of nanoparticles, and the distribution of the size,
s also important. Nanoparticles may behave differently according to
their size. At large sizes, nanoparticles or the agglomeration of
the particles may block blood vessels either of which can affect
the performance and safety of the composition.
[0113] Paclitaxel Derivatives
[0114] Paclitaxel derivatives used in the present invention
preferably are less toxic than paclitaxel. That is, the paclitaxel
derivative is less toxic to cells (e.g., does not kill cancer
and/or normal cells as well as, or does not inhibit cell
proliferation as well as) compared to paclitaxel. Paclitaxel
derivatives may include any derivative or precursor of paclitaxel.
Examples include, but are not limited to,
20-Acetoxy-4-deactyl-5-epi-20, O-secotaxol (the Meerwein Product of
Paclitaxel), or Baccatin III ((2.beta., 5.alpha., 7.alpha.,
10.alpha.,
13.beta.)-4,10-Diacetoxy-1,7,13-trihydroxy-9-oxo-5,20-epoxytax-11-en-2-yl
benzoate).
[0115] For example, most of the C-13 simplified paclitaxel
derivatives and derivatives of different stereochemistry
demonstrated reduced activity in comparison to paclitaxel. The
2'-hydroxyl and the 3'-benzamido group are not essential for
bioactivity, but are important for strong microtubule binding and
cytotoxicity. Formation of ethers at 2'-hydroxyl group, such as
alkyl ether (e.g., methyl ether) and alkyl silyl ether (e.g.,
ter-butyldimethylsilyl ether), reduced cytotoxicity. Acylation of
the 2'-hydroxyl group (e.g., 2'-acetyltaxol) also leads to loss of
activity. Replacement of 2'-hydroxyl group by halogen (e.g,
fluorine) was also found to significantly reduce the cytotoxicity.
Lead tetracetate oxidation of 6.alpha.-hydroxy-7-epi-paclitaxel
leads to C-nor-paclitaxel and C-seco-paclitaxel derivatives.
Tetrapropylammonium perruthenate (TPAP) oxidation of a
6.alpha.-hydroxy-7-epi-paclitaxel derivative leads to a
6-formyl-C-nor-paclitaxcel derivative. Reaction of a
6.alpha.-O-trifluoromethanesulfonyl-7-epi-paclitaxel derivative
with DMAP yields a
20-O-acetyl-4-deacetyl-5,6-dehydro-6-formyl-C-nor-paclitaxel
derivative. C-nor-paclitaxel analogs are less active than
paclitaxel. Therefore, the paclitaxel derivative may include
modifications at any one or more of these sites or produced by one
or more of these methods. See, e.g., Liang et al. Tetrahedron,
53(10):3441-3456 (1997); U.S. Pat. Nos. 5,756,301; and 5,981,777;
each of which is incorporated by reference herein in its
entirety.
Methods of Making Nahoparticles and Nanoparticle Complexes
[0116] In some aspects, the current invention relates to methods of
making nanoparticle compositions as described herein. In some
embodiments, this disclosure relates to methods of making the
nanoparticle compositions, wherein said method comprises contacting
the carrier protein and the paclitaxel or paclitaxel derivative
under conditions and ratios of components that will allow for
formation of the desired nanoparticles. In some embodiments, the
methods further relate to contacting a nanoparticle with binding
agents and/or therapeutic agent under conditions to form
nanoparticle complexes.
[0117] In some embodiments, the nanoparticles are made by combining
the carrier protein and paclitaxel or paclitaxel derivative, and
homogenizing under high pressure to form stable nanoparticles.
Non-limiting methods for homogenizing albumin and taxane
paclitaxel) can be found, for example, in Example 1, as well as
U.S. Pat. Nos. 5,916,596; 6,506,405; and 6,537,579, each of which
is incorporated herein by reference in its entirety.
[0118] In one embodiment, binding agents and/or therapeutic agents
are complexed to the nanoparticles as described, for example, in
U.S. Pat. Nos. 9,757,453, and 9,446,148.
[0119] In some aspects, provided herein are methods of making
nanoparticle complexes, wherein said methods comprise contacting
the carrier protein-paclitaxel (or paclitaxel derivative)
nanoparticle with the binding agents and/or therapeutic agent in a
solution haing a pH of between 5.0 and 7.5 and a temperature
between about 5.degree. C. and about 60.degree. C., between about
23.degree. C. and about 60.degree. C., or between about 55.degree.
C. and about 60.degree. C. under conditions and ratios of
components that will allow for formation of the desired
nanoparticles. In one embodiment, the nanoparticle is made at
55-60.degree. C. and pH 7.0. In another aspect provided herein are
methods of making the nanoparticle complexes, wherein said method
comprises (a) homogenizing the carrier protein and paclitaxel (or
paclitaxel derivative) to form a nanoparticle core; and (b)
contacting the core with the antibodies in a solution having a pH
of about 5.0 to about 7.5 at a temperature between about 5.degree.
C. and about 60.degree. C., between about 23.degree. C. and about
60.degree. C., or between about 55.degree. C. and about 60.degree.
C. under conditions and ratios of components that will allow for
formation of the desired nanoparticle complexes.
[0120] Another embodiment of the invention includes a method for
making a nanoparticle composition by facilitating complex formation
of a protein carrier and a therapeutic agent, the method comprising
contacting the carrier protein, an amount of paclitaxel as
described herein, and a therapeutic agent in a solution having a pH
of 5.0 or greater and a temperature between about 5.degree. C. and
about 60.degree. C., to generate a nanoparticle. In yet another
embodiment, the method further comprises contacting the
protein-carrier complex with a binding agent.
[0121] The amount of components (e.g., carrier protein, antibodies,
paclitaxel or paclitaxel derivative, therapeutic agents,
combinations thereof) is controlled in order to provide for
formation of the desired nanoparticles. A composition wherein the
amount of components is too dilute will not form the nanoparticles
as described herein. An overly concentrated solution will result in
unstructured aggregates.
[0122] In a preferred embodiment, weight ratio of carrier protein
to binding agent is 10:4. In some embodiments, the amount of carder
protein is between about 1 mg/mL and about 100 mg/mL. In some
embodiments, the amount of binding agent is between about 1 mg/mL
and about 30 mg/mL. For example, in some embodiments, the ratio of
carrier protein: binding agent: solution is approximately 9 mg of
carrier protein (e.g., albumin) to 4 mg of binding agent (e.g.,
BEV) in 1 mL of solution (e.g., saline).
[0123] In one embodiment, the amount of solution or other liquid
medium employed to form the nanoparticles is particularly
important. In some embodiments, the amount of solution (e.g.,
sterile water, saline, phosphate buffered saline) employed is
between about 0.5 mL of solution to about 20 mL of solution.
[0124] In one aspect, the nanoparticle complexes of the
nanoparticle composition are formed by contacting the carrier
protein-paclitaxel (opr paclitaxel derivative) nanoparticles and at
least one therapeutic agent with the binding agent at a ratio of
about 10:1 to about 10:30 carrier protein particle or carrier
protein-therapeutic agent particle to binding agent. In one
embodiment, the ratio is about 10:2 to about 10:25. In one
embodiment, the ratio is about 10:2 to about 1:1. In a preferred
embodiment, the ratio is about 10:2 to about 10:6. In an especially
preferred embodiment, the ratio is about 10:4. Contemplated ratios
include any value, subrange, or range within any of the recited
ranges, including endpoints.
[0125] In one aspect, the nanoparticles of the nanoparticle
composition are formed by contacting the carrier protein-paclitaxel
(or paclitaxel derivative) nanoparticles and at least one
therapeutic agent. In one embodiment, the ratio is about 10:2 to
about 10:25. In one embodiment, the ratio is about 10:2 to about
1:1. In a preferred embodiment, the ratio is about 10:2 to about
10:6. In an especially preferred embodiment, the ratio is about
10:4. Contemplated ratios include any value, subrange, or range
within any of the recited ranges, including endpoints.
[0126] In one embodiment, the carrier protein-paclitaxel (or
paclitaxel derivative) nanoparticles are contacted with the binding
agent in a solution having a pH between about 4 and about 8. In one
embodiment, the carrier protein-paclitaxel (or paclitaxel
derivative) nanoparticles are contacted with the binding agent in a
solution having a pH of about 4. In one embodiment, the carrier
protein-paclitaxel (or paclitaxel derivative) nanoparticles are
contacted with the binding agent in a solution having a pH of about
5. In one embodiment, the carrier protein-paclitaxel (or paclitaxel
derivative) nanoparticles are contacted with the binding agent in a
solution having a pH of about 6. In one embodiment, the carrier
protein-paclitaxel (or paclitaxel derivative) nanoparticles are
contacted with the binding agent in a solution having a pH of about
7. In one embodiment, the carrie carrier protein-paclitaxel (or
paclitaxel derivative) nanoparticles are contacted with the binding
agent in a solution having a pH of about 8. In a preferred
embodiment, the carrier protein-pad itaxel (or paclitaxel
derivative) nanoparticles are contacted with the binding agent in a
solution having a pH between about 5 and about 7.
[0127] In one embodiment, the carrier protein-paclitaxel (or
paclitaxel derivative) nanoparticles are incubated with the binding
agent at a temperature of about 5.degree. C. to about 60.degree.
C., or any range, subrange, or value within that range including
endpoints. In a preferred embodiment, the carrier
protein-paclitaxel (or paclitaxel derivative) nanoparticles are
incubated with the binding agent at a temperature of about
23.degree. C. to about 60.degree. C.
[0128] Without being bound by theory, it is believed that the
stability of the nanoparticle complexes is, at least in part,
dependent upon the temperature and/or pH at which the nanoparticles
are formed, as well as the concentration of the components carrier
protein, binding agent, paclitaxel or paclitaxel derivative, and a
therapeutic agent) in the solution. In one embodiment, the K.sub.d
of the nanoparticles is between about 1.times.10.sup.-11 M and
about 2.times.10.sup.-5 M. In one embodiment, the K.sub.d of the
nanoparticles is between about 1.times.10.sup.-11 M and about
2.times.10.sup.-8 M. In one embodiment, the K.sub.d of the
nanoparticles is between about 1.times.10.sup.-11 M and about
7.times.10.sup.-9 M. In a preferred embodiment, the K.sub.d of the
nanoparticles is between about 1.times.10.sup.-11 M and about
3.times.10.sup.-8 M. Contemplated values include any value,
subrange, or range within any of the recited ranges, including
endpoints.
Making Reduced-Toxicity Nanoparticles
[0129] The relative weight ratio of the carrier protein to the
paclitaxel is greater than about 9:1. In one embodiment, the
earlier protein and the paclitaxel have a relative weight ratios of
greater than about 10:1, or about 11:1, or about 12:1, or about
13:1, or about 14:1, or about 15:1, or about 16:1, or about 17:1,
or about 18:1, or about 19:1, or about 20:1, or about 21:1, or
about 22:1, or about 23:1, or about 24:1, or about 25:1, or about
26:1, or about 27:1, or about 28:1, or about 29:1, or about 30:1,
in one embodiment, the weight ratio is between 10:1 and 100:1,
between 10:1 and 50:1, between 10:1 and 40:1, between 10:1 and
30:1, between 10:1 and 20:1, or between 10:1 and 15:1. In one
embodiment, the weight ratio is between 15:1 and 50:1. 15:1 and
50:1, between 15:1 and 40:1, between 15:1 and 30:1, or between 15:1
and 20:1. The weight ratio may be any value or subrange within the
recited ranges, including endpoints.
[0130] In one embodiment, the amount of paclitaxel is equal to a
minimum amount capable of providing stability to the nanoparticles.
In one embodiment, the amount of paclitaxel is greater than or
equal to a minimum amount capable of providing affinity of the at
least one therapeutic agent to the protein carrier. In one
embodiment, the amount of paclitaxel is greater than or equal to a
minimum amount capable of facilitating complex formation of the at
least one therapeutic agent and the protein carrier. For example,
less than about I mg of taxol can be added 9 mg of carrier protein
(10 mg carrier protein-therapeutic) and 4 mg of binding agent,
e.g., antibody, Fc fusion molecule, or aptamer, in 1 ml of
solution.
[0131] It is to be noted that, when using a typical i.v, bag, for
example, with the solution of approximately 1 liter one would need
to use 1000.times. the amount of carrier protein/carrier protein
therapeutic agent and antibodies compared to that used in 1 mL.
Thus, one cannot form the present nanoparticles in a standard i.v.
bag. Furthermore, when the components are added to a standard i.v.
bag in the therapeutic amounts of the present invention, the
components do not self-assemble to form nanoparticles.
[0132] In one embodiment, the amount of paclitaxel present in the
nanoparticle composition is less than about 5 mg/mL. In one
embodiment, the amount of paclitaxel present in the nanoparticle
composition is less than about 4.54 mg/mL, or about 4.16 mg/mL, or
about 3.57 mg/mL, or about 3.33 mg/mL, or about 3.12 mg/mL, or
about 2.94 mg/mL, or about 2.78 mg/mL, or about 2.63 mg/mL, or
about 2.5 mg/mL, or about 2.38 mg/mL, or about 2.27 mg/mL, or about
2.17 mg/mL, or about 2.08 mg/mL, or about 2 mg mL, or about 1.92
mg/mL, or about 1.85 mg/mL, or about 1.78 mg/mL, or about 1.72
mg/mL, or about 1.67 mg/mL. In one embodiment, the amount. of
paclitaxel present in the nanoparticle composition is greater than
or equal to a minimum amount capable of providing stability to the
nanoparticles. In one embodiment, the amount of paclitaxel present
in the nanoparticle composition is greater than or equal to a
minimum amount capable of providing affinity of the at least one
therapeutic agent to the protein carrier. In one embodiment, the
amount of paclitaxel present in the nanoparticle composition is
greater than or equal to a minimum amount capable of facilitating
complex formation of the at least one therapeutic agent and the
protein carrier.
[0133] In one embodiment, the nanoparticle compositions described
herein includes no binding agents.
Lyophilization
[0134] While protein compositions comprising a single source
protein are commonly stored in lyophilized form where they exhibit
significant shelf-life, such lyophilized compositions do not
contain a self-assembled nanoparticle of two different proteins
integrated together by hydrophobic-hydrophobic interactions.
Moreover, the nanoparticle configuration wherein a majority of the
binding portions of the binding agent are exposed on the surface of
the nanoparticles lends itself to being susceptible to dislodgement
or reconfiguration by conditions which otherwise would be
considered benign. For example, during lyophilization, ionic
charges on the proteins are dehydrated thereby exposing the
underlying charges. Exposed charges allow for charge-charge
interactions between the two proteins which can alter the binding
affinity of each protein to the other. In addition, the
concentration of the nanoparticles increases significantly as the
solvent (e.g., water) is removed. Such increased concentrations of
nanoparticles could lead to irreversible oligomerization.
Oligomerization is a known property of proteins that reduces the
biological properties of the oligomer as compared to the monomeric
form and increases the size of the particle sometimes beyond 1
micron.
[0135] On the other hand, a stable form of a nanoparticle
composition is required for clinical and/or commercial use where a
shelf-life of at least 3 months is required and shelf-lives of
greater than 6 months or 9 months are preferred. Such a stable
composition must be readily available for intravenous injection,
must retain its self-assembled form upon intravenous injection so
as to direct the nanoparticle to the predetermined site in vivo,
must have a maximum size of less than 1 micron so as to avoid any
ischemic event when delivered into the blood stream, and finally
must be compatible with the aqueous composition used for
injection.
[0136] Lyophilization, or freeze-drying, removes water from a
composition. In the process, the material to be dried is first
frozen and then the ice or frozen solvent is removed by sublimation
in a vacuum environment. An excipient may be included in
pre-lyophilized formulations to enhance stability during the
freeze-drying process and/or to improve stability of the
lyophilized product upon storage. Pikal, M. Biopharm. 3(9)26-30
(1990) and Arakawa et al., Pharm. Res. 8(3): 285-291 (1991).
[0137] While proteins may be lyophilized, the process of
lyophilization and reconstitution may affect the properties of the
protein. Because proteins are larger and more complex than
traditional organic and inorganic drugs (i.e. possessing multiple
functional groups in addition to complex three-dimensional
structures), the formulation of such proteins poses special
problems. For a protein to remain biologically active, a
formulation must preserve intact the conformational integrity of at
least a core sequence of the protein's amino acids while at the
same time protecting the protein's multiple functional groups from
degradation. Degradation pathways for proteins can involve chemical
instability (i.e. any process which involves modification of the
protein by bond formation or cleavage resulting in a new chemical
entity) or physical instability (i.e. changes in the higher order
structure of the protein). Chemical instability can result from
deamidation, racemization, hydrolysis, oxidation, beta elimination
or disulfide exchange. Physical instability can result from
denaturation, aggregation, precipitation or adsorption, for
example. The three most common protein degradation pathways are
protein aggregation, deamidation and oxidation. Cleland, et al.,
Critical Reviews in Therapeutic Drug Carrier Systems 10(4): 307-377
(1993).
[0138] Finally, cryoprotectants and agents that assist in the
lyophilization process must be safe and tolerated for therapeutic
use.
[0139] The lyophilized compositions of this invention are prepared
by standard lyophilization techniques with or without the presence
of stabilizers, buffers, etc. Surprisingly, these conditions do not
alter the relatively fragile structure of the nanoparticles.
Moreover, at best, these nanoparticles retain their size
distribution upon lyophilization and, more importantly, can be
reconstituted for in vivo administration (e.g., intravenous
delivery) in substantially the same form and ratios as if freshly
made.
[0140] In some embodiments, the nanoparticles RTP or NTP) are
lyophilized. In some embodiments, the nanoparticle complexes (e.g.,
RTP or NTP with associated therapeutic agent and/or binding agents)
are lyophilized. In some embodiments, upon reconstitution with an
aqueous solution, an amount of the binding agents are arranged on a
surface of the nanoparticles. In some einbodimetns, the
nanoparticle complexes are capable of binding to an antigen. In
some embodiments, the antibodies associated with the nanoparticle
complexes remain capable of binding to the antigen.
Formulations
[0141] In one aspect, the nanoparticle composition is formulated
for systemic delivery, e.g., intravenous administration.
[0142] In one aspect, the nanoparticle composition is formulated
for direct injection into a tumor. Direct injection includes
injection into or proximal to a tumor site, perfusion into a tumor,
and the like. Because the nanoparticle composition is not
administered systemically, a nanoparticle composition is formulated
for direct injection into a tumor may comprise any average particle
size. Without being bound by theory, it is believed that larger
particles (e.g., greater than 500 nm, greater than 1 .mu.m, and the
like) are more likely to be immobilized within the tumor, thereby
providing what is believed to be a better beneficial effect.
[0143] In another aspect, provided herein is a composition
comprising a compound provided herein, and at least one
pharmaceutically acceptable excipient.
[0144] In general, the compounds provided herein can be formulated
for administration to a patient by any of the accepted modes of
administration. Various formulations and drug delivery systems are
available in the art. See, e.g., Gennaro, A. R., ed. (1995)
Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing
Co.
[0145] In general, compounds provided herein will be administered
as pharmaceutical compositions by any one of the following routes:
oral, systemic (e.g., transdermal, intranasal or by suppository),
or parenteral (e.g., intramuscular, intravenous or subcutaneous)
administration.
[0146] The compositions are comprised of, in general, a. compound
of the present invention in combination with at least one
pharmaceutically acceptable excipient. Acceptable excipients are
non-toxic, aid administration, and do riot adversely affect the
therapeutic benefit of the claimed compounds. Such excipient may be
any solid, liquid, semi-solid or, in the case of an aerosol
composition, gaseous excipient that is generally available to one
of skill in the art.
[0147] Solid pharmaceutical excipients include starch, cellulose,
talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk,
silica gel, magnesium stearate, sodium stearate, glycerol
monostearate, sodium chloride, dried skim milk and the like. Liquid
and semisolid excipients may be selected from glycerol, propylene
glycol, water, ethanol and various oils, including those of
petroleum, animal, vegetable or synthetic origin, e.g., peanut oil,
soybean oil, mineral oil, sesame oil, etc. Preferred liquid
carriers, particularly for injectable solutions, include water,
saline, aqueous dextrose, and glycols. Other suitable
pharmaceutical excipients and their formulations are described in
Remington.'s Pharmaceutical Sciences, edited by E. W. Martin (Mack
Publishing Company, 18th ed., 1990).
[0148] The present compositions may, if desired, be presented in a
pack or dispenser device containing one or more unit dosage forms
containing the active ingredient. Such a pack or device may, for
example, comprise metal or plastic foil, such as a blister pack, or
glass, and rubber stoppers such as in vials. The pack or dispenser
device may be accompanied by instructions for administration.
Compositions comprising a compound of the invention formulated in a
compatible pharmaceutical carrier may also be prepared, placed in
an appropriate container, and labeled for treatment of an indicated
condition.
Treatment Methods
[0149] The nanoparticle compositions as described herein are useful
in treating cancer cells and/ or tumors in a mammal. In a preferred
embodiment, the mammal is a human a human patient). Preferably, a
lyophilized nanoparticle composition is reconstituted (suspended in
an aqueous excipient) prior to administration.
[0150] In one aspect is provided a method for treating a cancer
cell, the method comprising contacting the cell with an effective
amount of nanoparticle composition as described herein to treat the
cancer cell. Treatment of a cancer cell includes, without
limitation, reduction in proliferation, killing the cell,
preventing metastasis of the cell, and the like.
[0151] In one aspect is provided a method for treating a tumor in a
patient in need thereof, the method comprising administering to the
patient a therapeutically effective amount of a nanoparticle
composition as described herein to treat the tumor. In one
embodiment, where the nanoparticles include a binding agent
comprising an antigen-binding domain, the method comprises
selecting a patient having a cancer which expresses the antigen. In
one embodiment, the size of the tumor is reduced. In one
embodiment, the tumor size does not increase (i.e. progress) for at
least a period of time during and/or after treatment.
[0152] In one embodiment, the nanoparticle composition is
administered intravenously. In one embodiment, the n.anoparticle
composition is administered directly to the tumor. In one
embodiment, the nanoparticle composition is administered by direct
injection or perfusion into the tumor.
[0153] In one embodiment, the method comprises: [0154] a)
administering the nanoparticle composition once a week for three
weeks; [0155] b) ceasing administration of the nanoparticle
composition for one week; and [0156] c) optionally repeating steps
a) and b) as necessary to treat the tumor.
[0157] In one embodiment, the therapeutically effective amount of
the nanoparticles described herein comprises about 1 mg/m.sup.2 to
about 200 mg/m.sup.2 antibody, about 2 mg/m.sup.2 to about 150
mg/m.sup.2, about 5 mg/m.sup.2 to about 100 mg/m.sup.2, about 10
mg/m.sup.2 to about 85 mg/m.sup.2, about 15 mg/m.sup.2 to about 75
mg/m.sup.2, about 20 mg/m.sup.2 to about 65 mg/m.sup.2, about 25
mg/m.sup.2 to about 55 mg/m.sup.2, about 30 mg/m.sup.2 to about 45
mg/m.sup.2, or about 35 mg/m.sup.2 to about 40 mg/m.sup.2 antibody.
In other embodiments, the therapeutically effective amount
comprises about 20 mg/m.sup.2 to about 90 mg/m.sup.2 antibody. In
one embodiment, the therapeutically effective amount comprises 30
mg/m.sup.2 to about 70 mg/m.sup.2 antibody.
[0158] In one embodiment, the therapeutically effective amount of
the nanoparticles described herein comprises about 50 mg/m.sup.2 to
about 200 mg/mn.sup.2 carrier protein or carrier protein and
therapeutic agent. In one embodiment, the therapeutically effective
amount of the nanoparticle compositions described herein comprise
no binding agents. In one embodiment, the therapeutically effective
amount of the nanoparticle compositions described herein comprise
less than about a therapeutically effective amount of paclitaxel.
In a preferred embodiment, the therapeutically effective amount
comprises about less than 75 mg/m.sup.2 of paclitaxel. Contemplated
values include any value, subrange, or range within any of the
recited ranges, including endpoints.
[0159] In one embodiment, the therapeutically effective amount
comprises about 20 mg/m.sup.2 to about 90 mg/m.sup.2 binding agent,
e.g., antibody, aptamer or fusion protein. In a preferred
embodiment, the therapeutically effective amount comprises 30
mg/m.sup.2 to about 70 mg/m.sup.2 binding agent, e.g., antibody,
aptamer or fusion protein. Contemplated values include any value,
subrange, or range within any of the recited ranges, including
endpoints.
[0160] Cancers or tumors that can be treated by the compositions
and methods described herein include, but are not limited to the
cancers referred to in the above tables, as well as: biliary tract
cancer, brain cancer, including glioblastomas and medulloblastomas;
breast cancer; cervical cancer; choriocarcinoma; colon cancer;
endometrial cancer; esophageal cancer, gastric cancer;
hematological neoplasms, including acute lymphocytic and
myelogenous leukemia; multiple myeloma; AIDS associated leukemias
and adult T-cell leukemia lymphoma; intraepithelial neoplasms,
including Bowen's disease and Paget's disease; liver cancer
(hepatocarcinoma); lung cancer; lymphoma.s., including Hodgkin's
disease and lymphocytic lymphomas; neuroblastomas; oral cancer,
including squamous cell carcinoma; ovarian cancer, including those
arising from epithelial cells, stromal cells, germ cells and
mesenchymal cells; pancreas cancer; prostate cancer; rectal cancer;
sarcomas, including leiomyosarcoma, rhabdomyosarcoma, liposarcoma,
fibrosarcoma. and osteosarcoma; skin cancer, including melanoma,
Kaposi's sarcoma, basocellular cancer and squamous cell cancer;
testicular cancer, including germinal tumors (serninorna,
non-seminomatteratomas, choriocarcinomas), stromal tumors and germ
cell tumors; thyroid cancer, including thyroid adenocarcinoma and
medullar carcinoma; and renal cancer including adenocarcinoma and
Wilms tumor. In important embodiments, cancers or tumors include
breast cancer, lymphoma, multiple myelorna, and melanoma.
[0161] In general, the compounds of this invention will be
administered in a therapeutically effective amount by any of the
accepted modes of administration for agents that serve similar
utilities. The actual amount of the compound of this invention,
i.e., the nanoparticles, depend upon numerous factors such as the
severity of the disease to be treated, the age and relative health
of the subject, the potency of the compound used, the route and
form of administration, and other factors well known to the skilled
artisan.
[0162] An effective amount of such agents can readily be determined
by routine experimentation, as can the most effective and
convenient route of administration, and the most appropriate
formulation. Various formulations and drug delivery systems are
available in the art. See, e.g., Gennaro, A. R., ed. (1995)
Remington's Pharmaeutical Sciences, 18th ed., Mack Publishing
Co.
[0163] An effective amount or a therapeutically effective amount or
dose of an agent, e.g., a compound of the invention, refers to that
amount of the agent or compound that results in amelioration of
symptoms or a prolongation of survival in a subject. Toxicity and
therapeutic efficacy of such molecules can be determined by
standard pharmaceutical procedures in cell cultures or experimental
animals, e.g., by determining the LD50 (the dose lethal to 50% of
the population) and the ED50 (the dose therapeutically effective in
50% of the population). The dose ratio of toxic to therapeutic
effects is the therapeutic index, which can be expressed as the
ratio LD50/ED50. Agents that exhibit high therapeutic indices are
preferred.
[0164] The effective amount or therapeutically effective amount is
the amount of the compound or pharmaceutical composition that will
elicit the biological or medical response of a tissue, system,
animal or human that is being sought by the researcher,
veterinarian, medical doctor or other clinician. Dosages may vary
within this range depending upon the dosage form employed and/or
the route of administration utilized. The exact formulation, route
of administration, dosage, and dosage interval should be chosen
according to methods known in the art, in view of the specifics of
a subject's condition.
[0165] Dosage amount and interval may be adjusted individually to
provide plasma levels of the active moiety that are sufficient to
achieve the desired effects; i.e., the minimal effective
concentration (MEC). The MEC will vary for each compound but can be
estimated from, for example, in vitro data and animal experiments.
Dosages necessary to achieve the MEC will depend on individual
characteristics and route of administration. In cases of local
administration or selective uptake, the effective local
concentration of the drug may not be related to plasma
concentration.
[0166] In related embodiments, the treatment comprises
administration of the targeting binding agent prior to
administration of the nanoparticles. In one embodiment, the
targeting binding agent is administered between about 6 and 48, or
12 and 48 hours prior to administration of the nanoparticles. In
another embodiment, the targeting binding agent is administered
between 6 and 12 hours prior to administration of the
nanopatticles. In yet another embodiment, the targeting binding
agent is administered between 2 and 8 hours prior to administration
of the nanoparticles. In still other embodiments, the targeting
binding agent is administered a week prior to administration of the
nanoparticles. For example, administration of a dose of BEV 24
hours prior to administration of nanopa.rticle complexes comprising
bevacizumab or other VEGF-binding antibody. In another example,
prior administration of rituximab prior to administering
rituximab-containing nanoparticle complexes. The binding agent
administered prior to the nanoparticle may be administered as a
dose that is subtherapeutic, such as 1/2, 1/10th or 1/20 the amount
normally considered therapeutic. Thus, in humans, pretreatment with
BEV may comprise administration of ling/kg BEV which is 1/10 th the
usual dose, followed by administration of the nanoparticles or
nanoparticle complexes.
EXAMPLES
[0167] The present disclosure is illustrated using nanoparticles
composed of albumin-bound paclitaxel or paclitaxel derivative as a
core, and bevacizumab (i.e., Avastin.RTM.) or Rituximab (i.e.,
Rituxang) as antibodies.
[0168] One skilled in the art would understand that making and
using the nanoparticles of the Examples are for the sole purpose of
illustration, and that the present disclosure is not limited by
this illustration.
[0169] Any abbreviation used herein, has normal scientific meaning.
All temperatures are .degree. C. unless otherwise stated. Herein,
the following terms have the following meanings unless otherwise
defined:
TABLE-US-00003 ABX = ABRAXANE .RTM./(albumin-bound paclitaxel ADC =
antibody dependent chemotherapy BEV = bevacizumab BSA = bovine
serum albumin nM = nanomolar FITC = Fluorescein Kd = dissociation
constant kg = kilogram M = molar rug = milligram ml or mL =
milliliter m.sup.2 = square meters mm.sup.3 = cubic millimeter
.mu.g = microgram .mu.l = microliter .mu.m = micrometer/micron PBS
= Phosphate buffered saline RT = room temperate rpm = rotations per
minute
Example 1: Nanoparticle Preparation
[0170] Paclitaxel was reacted with Meerwein's Reagent breaking
according to die reaction scheme laid out by Samaranayake et al. in
"Modified Taxols. Reaction of Taxol with Electrophilic Reagents and
Preperation of a Rearranged Taxol Derivative with Tubulin Assembly
Activity" (J. Org. Chem, 1991, 56, 5114-5119), which is
incorporated herein by reference in its entirety. This reaction
breaks the C-4,C-5 oxetane ring severely inhibiting toxicity. The
reaction and purification were performed by Organix Inc. (Woburn,
Mass.). The reaction is shown in FIG. 1.
[0171] The Meerwein's Product of Paclitaxel
(20-Acetoxy-4-deactyl-5-epi-20, O-secotaxol) was then homogenized
under high pressure with albumin to form stable non-toxic
nanoparticles (NTP). Methods for homogenizing albumin and taxane
(e.g., paclitaxel) can be found, for example, in U.S. Pat. Nos.
5,916,596; 6,506,405; and 6,537,579, each of which is incorporated
herein by reference in its entirety.
[0172] Briefly, human serum albumin (HSA; Sigma Aldrich) was
dissolved in sterile water at 1-1.5% w/v. In a separate container,
20-Acetoxy-4-deactyl-5-epi-20, O-secotaxol (10% w/w to HSA) was
dissolved in chloroform to 10 mg/ml. Chloroform solution was added
to the albumin solution and mixed with a hand-held homogenizer for
5 minutes at low RPM. The solution was run through
micro-homogenizing pump at 20,000 PSI for 15 cycles. Pump coiled
pathway and collection tube were placed in an ice bath to prevent
solution from reaching dangerously high temps (i.e. over 50.degree.
C.). The solution was transferred to a rotary evaporator with a
cold trap and run at reduced pressure for 45 minutes to evaporate
off chloroform. The solution was filtered through 200 nm cellulose
acetate filter.
[0173] For lyophilized nanoparticles, the solution was split into
freeze drying flasks at appropriate volumes to allow full
lyophilization of product. Flask with solution were placed. at
-80.degree. C. for 2-4 hours. The frozen solution in the flasks
were lyophilized using a bench top freeze dryer with condenser of
-85.degree. C. at <20 mtorr.
Example 2: Binding of NTP and Bevacizumab In Vitro
[0174] To determine whether NTP and bevacizumba (BEV) interact, the
size of the NTP formed in Example 1 was analyzed by Malvern
Nanosight technology, alone and after incubatluon with 4, 6, 8, and
10 miligrams per milliliter (mg/mL) of bevacizumab for 30
minutes.
[0175] Increasing nanoparticle diameter with increased
concentration of BEV (FIGS. 2A-2E) suggests increased non-covalent
coating of NTP with BEV. Diameter was measured at a 1:300 dilution
using Malvern Nanosight technology.
Example 3: Affinity of Bevacizumab and Rituximab Binding to
Non-Toxic Nanoparticles (NTP)
[0176] Binding affinity (K.sub.d) of both bevacizumab (FIG. 3A) and
rituximab (FIG. 3B) to NTP was measured using Bio Layer
Interferometry Technology. Both antibodies were biotinylated using
amine coupling and then bound individually to a BLITZ Streptavidin
probe at 100 .mu.g/mL. NTP at various concentrations was assayed
for binding against the probe with association and dissociation
periods of 120 seconds. Kinetic analysis was performed by BLITZ
Evaluation software. Nano-picomolar affinities indicate strong
binding between rituximab and NTP, and bevaciuzmab and NTP.
Example 4: Stability of Non-Toxic Nanoparticles (NTP) in PBS and
Serum Compared to ABRAXANE.RTM. (ABX)
[0177] Antibody free NTP and ABRAXANTA (ABX) solutions were made to
30 ng/nil. of Meerwein's Product of Paclitaxel and paclitaxel,
respectively, in PBS, A baseline count of nanoparticles per mL was
obtained using Malvern Nanosight technology. Then a range of
dilutions was made for both solutions, and particle concentration
was measured.
[0178] FIG. 4A shows the stability of ABX and NTP in saline.
Abraxane nanoparticles began to fall apart around 1:1.33 dilution
of initial solution until it reached the baseline PBS levels at
.about.1:2 dilution. NTP decreased linearly with dilution factor,
suggesting no particle fall apart.
[0179] FIG. 4B shows the stability of ABX and NTP in serum. NTP and
ABX solutions were made to 250 .mu.g/mL of Meerwein's Product of
Paclitaxel and paclitaxel, respectively, in serum. Initial
concentrations of nanoparticles were measured using Malvern
Nanosight Technology, subsequent measurements are presented as a
percentage of initial concentration. Concentrations of each
solution were measured after 15, 30, 45, and 60 minutes in
smut.
Example 5: Daudi Cell CD20 Blocking by NTP-Ritoximab Complexes
[0180] ABX, AR160, Non-toxic nanoparticle (NTP) and rituximab-bound
NTP were prepared as described above or previously (see, e.g., PCT
Pub. No. WO2014/055415, which is incorporated herein by reference
in its entirety). The nanoparticles and unlabeled rituximab were
co-incubated with CD20+ Daudi lymphoma cells for 30 minutes at room
temperature. The cells were washed and subsequently stained with PE
anti-human CD20. Isotype (FIG. 5A) and anti-CD20 (FIG. 5B) stained
cells served as negative and positive controls, respectively.
[0181] The results indicate the AR160 (FIG. 5D), rituximab loaded
NT particles (NTRit; FIG. 5F), and rituximab (FIG. 5G) block
subsequent binding of PE anti-human CD20, while ABX (FIG. 5C) and
the NT nanoparticle (FIG. 5E) alone do not. This shows that the the
rituximab in the context of AR160 and rituximab-loaded NT particles
retains its ligand binding properties.
Example 6: Albumin-Paclitaxel Nanoparticles with Reduced
Paclitaxel
[0182] Albumin-paclitaxel nanoparticles having reduced amounts of
paclitaxel compared to ABRAXANE.RTM. are formed.
[0183] For example, less than 30 mg (e.g., 1 mg to 25 mg)
paclitaxel is dissolved in a solvent (e.g., methylene chloride; or
chloroform and ethanol). The solution is added to 27.0 mL of human
serum abumin solution (1% w/v). The mixture is homogenized for 5
minutes at low RPM in order to form a crude emulsion, and then
transferred into a high pressure homogenizer. The emulsification is
performed at 9000-40,000 psi while recycling the emulsion for at
least 5 cycles. The resulting system is transferred into a rotary
evaporator, and methylene chloride is rapidly removed at 40.degree.
C., at reduced pressure (30 mm Hg), for 20-30 minutes. Optionally,
the dispersion is filtered through a 0.22 micron filter.
[0184] The filtered nanoparticles may be lyophilized and
stored.
Example 7: Albumin-Paclitaxel-Therapeutic Agent Nanoparticles with
Reduced Paclitaxel
[0185] Albumin-paclitaxel-therapeutic agent nanoparticles having
reduced amounts of paclitaxel compared to ABRAXANE.RTM. are
formed.
[0186] For example, albumin-paclitaxel nanoparticles are formed as
described in Example 6. A therapeutic agent (e.g., cisplatin) is
incubated with the nanoparticles at a ratio of about 10:1
(nanoparticles:agent) at room temperature for 30 min. Free
therapeutic agent (e.g., cisplatin) is measured by HPLC in the
supernatant after nanoparticle particulate is removed.
Example 8: Albumin-Paclitaxel-Antibody Nanoparticles with Reduced
Paclitaxel
[0187] Albumin-paclitaxel-antbody nanoparticles having reduced
amounts of paclitaxel compared to ABRAXANE.RTM. are formed.
[0188] For example, albumin-paclitaxel nanoparticles are formed as
described in Example 6. Nanoparticles (10 mg) are combined with an
antibody (e.g., bevacizumab) (4 mg), and 840 .mu.l of 0.9% saline
is added to give a final concentration of 10 mg/ml and 2 mg/ml of
nanoparticles and antibody, respectively. The mixture is incubated
for 30 minutes at room temperature to allow particle formation.
[0189] An antibody-albumin-paclitaxel-therapeutic agent
nanoparticle complex may be formed. Optionally, antibody is
incubated with albumin-paclitaxel-therapeutic agent nanoparticles
(from Example 7) to form antibody-albumin-paclitaxel-therapeutic
agent nanoparticle complexes. Alternatively, the
antibody-albumin-paclitaxel nanoparticles my beincubated with the
therapeutic agent to form antibody-albumin-paclitaxel-therapeutic
agent nanoparticle complexes.
* * * * *