U.S. patent application number 16/948537 was filed with the patent office on 2021-02-04 for method for treating cancer using hsp90 inhibitors.
The applicant listed for this patent is Al Therapeutics, Inc.. Invention is credited to Neil Beeharry, Marylens Hernandez, Sean Landrette, Henri Lichenstein, Jonathan M. Rothberg, Tian Xu.
Application Number | 20210030759 16/948537 |
Document ID | / |
Family ID | 1000005164068 |
Filed Date | 2021-02-04 |
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United States Patent
Application |
20210030759 |
Kind Code |
A1 |
Beeharry; Neil ; et
al. |
February 4, 2021 |
METHOD FOR TREATING CANCER USING HSP90 INHIBITORS
Abstract
The disclosure also provides compositions and methods related to
combination therapy with HSP90 inhibitors and BCL-2 pathway
inhibitors for treating cancer. The disclosure also provides
compositions and methods related to the use of `low dose` HSP90
inhibitors in the treatment of cancer, alone and in combination
with other therapeutic agents.
Inventors: |
Beeharry; Neil; (Guilford,
CT) ; Hernandez; Marylens; (Guilford, CT) ;
Landrette; Sean; (Meriden, CT) ; Xu; Tian;
(Shanghai, CN) ; Rothberg; Jonathan M.; (Guilford,
CT) ; Lichenstein; Henri; (Guilford, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Al Therapeutics, Inc. |
Guilford |
CT |
US |
|
|
Family ID: |
1000005164068 |
Appl. No.: |
16/948537 |
Filed: |
September 23, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15885975 |
Feb 1, 2018 |
10799508 |
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16948537 |
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62587886 |
Nov 17, 2017 |
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62563991 |
Sep 27, 2017 |
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62454113 |
Feb 3, 2017 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/02 20180101;
A61K 31/395 20130101; A61K 31/437 20130101; A61K 45/06 20130101;
A61P 35/00 20180101; A61K 31/444 20130101; A61K 2300/00 20130101;
A61K 31/496 20130101; A61K 31/46 20130101; A61K 31/416 20130101;
A61K 31/52 20130101; A61K 31/635 20130101 |
International
Class: |
A61K 31/52 20060101
A61K031/52; A61K 31/496 20060101 A61K031/496; A61K 31/437 20060101
A61K031/437; A61K 31/46 20060101 A61K031/46; A61P 35/02 20060101
A61P035/02; A61K 31/416 20060101 A61K031/416; A61P 35/00 20060101
A61P035/00; A61K 31/444 20060101 A61K031/444; A61K 31/635 20060101
A61K031/635; A61K 31/395 20060101 A61K031/395; A61K 45/06 20060101
A61K045/06 |
Claims
1. A pharmaceutical composition comprising an HSP90 inhibitor and a
BCL-2 inhibitor, wherein the BCL-2 inhibitor is venetoclax and the
HSP90 inhibitor is selected from MPC-3100, MPC-0767, AT-13387,
tanespimycin, TAS-116, SNX-5422, XL-888, and pharmaceutically
acceptable salts thereof, and a pharmaceutically acceptable carrier
or excipient.
2. The pharmaceutical composition of claim 1, wherein the HSP90
inhibitor is selected from WC-3100, MPC-0767, and pharmaceutically
acceptable salts thereof.
3. A method for treating a hematopoietic or lymphoid cancer
selected from a leukemia, a lymphoma and a multiple myeloma in a
subject in need thereof, the method comprising administering to the
subject the pharmaceutical composition of claim 1 or claim 2.
4. A method for treating a hematopoietic or lymphoid cancer
selected from a leukemia, a lymphoma and a multiple myeloma in a
subject in need thereof, the method comprising administering to the
subject an HSP90 inhibitor and a BCL-2 inhibitor, wherein the BCL-2
inhibitor is venetoclax and the HSP90 inhibitor is MPC-3100, or a
pharmaceutically acceptable salt thereof.
5. The method of claim 3, wherein the cancer is characterized as
positive for BCL-2 expression based on the expression of BCL-2 in a
biological sample of the cancer.
6. The method of claim 4, wherein the cancer is characterized as
positive for BCL-2 expression based on the expression of BCL-2 in a
biological sample of the cancer.
7. The method of claim 5, wherein the cancer characterized as
positive for BCL-2 expression is a cancer in which a biological
sample from the cancer expresses BCL-2 at a level that is at least
two-fold higher compared to the BCL-2 expression in a reference
sample of non-cancerous tissue.
8. The method of claim 6, wherein the cancer characterized as
positive for BCL-2 expression is a cancer in which a biological
sample from the cancer expresses BCL-2 at a level that is at least
two-fold higher compared to the BCL-2 expression in a reference
sample of non-cancerous tissue.
9. The method of claim 5, wherein the BCL-2 expression is protein
expression or gene expression.
10. The method of claim 6, wherein the BCL-2 expression is protein
expression or gene expression.
11. The method of claim 3, wherein the HSP90 inhibitor is
administered at a dose that is less than 90% of the recommended
phase 2 dose of the HSP90 inhibitor.
12. The method of claim 4, wherein the HSP90 inhibitor is
administered at a dose that is less than 90% of the recommended
phase 2 dose of the HSP90 inhibitor.
13. The method of claim 3, wherein the cancer is a leukemia
selected from acute lymphoblastic leukemia (ALL), acute myelogenous
leukemia (AML), chronic lymphocytic leukemia (CLL), chronic
myelogenous leukemia (CML), and acute monocytic leukemia.
14. The method of claim 4, wherein the cancer is a leukemia
selected from acute lymphoblastic leukemia (ALL), acute myelogenous
leukemia (AML), chronic lymphocytic leukemia (CLL), chronic
myelogenous leukemia (CML), and acute monocytic leukemia.
15. The method of claim 13, wherein the cancer is AML.
16. The method of claim 14, wherein the cancer is AML.
17. The method of claim 3, wherein the cancer is a lymphoma
selected from a Hodgkins and a Non-Hodgkin's lymphoma.
18. The method of claim 4, wherein the cancer is a lymphoma
selected from a Hodgkins and a Non-Hodgkin's lymphoma.
19. The method of claim 17, wherein the cancer is a Non-Hodgkin's B
cell lymphoma, preferably selected from a diffuse large B cell
lymphoma (DLBCL), Burkitt lymphoma, lymphoblastic lymphoma, and
mantle cell lymphoma, and most preferably selected from a diffuse
large B cell lymphoma (DLBCL) and a mantle cell lymphoma.
20. The method of claim 18, wherein the cancer is a Non-Hodgkin's B
cell lymphoma, preferably selected from a diffuse large B cell
lymphoma (DLBCL), Burkitt lymphoma, lymphoblastic lymphoma, and
mantle cell lymphoma, and most preferably selected from a diffuse
large B cell lymphoma (DLBCL) and a mantle cell lymphoma.
21. The method of claim 3, wherein the cancer is a multiple
myeloma.
22. The method of claim 4, wherein the cancer is a multiple
myeloma.
23. The method of claim 3, wherein the subject is human.
24. The method of claim 4, wherein the subject is human.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 15/885,975, filed on Feb. 1, 2018, which
claims priority to U.S. Provisional Patent Application No.
62/587,886 filed on Nov. 17, 2017, U.S. Provisional Patent
Application No. 62/563,991 filed on Sep. 27, 2017, and U.S.
Provisional Patent Application No. 62/454,113 filed on Feb. 3,
2017, the contents of which are hereby fully incorporated by
reference.
FIELD OF THE INVENTION
[0002] The invention relates to the use of HSP90 inhibitors to
modulate the immune system for the treatment of cancer and to their
use in combination with inhibitors of BCL-2 for treating
cancer.
BACKGROUND OF THE INVENTION
[0003] Approaches to harness the immune system to effectively
identify and subsequently eradicate tumor cells were discovered
more than a decade ago. It is only through the recent generation of
checkpoint antibodies, notably those that target the programmed
cell death protein (PD-1)/programmed death-ligand 1 (PD-L1), has
the potential been translated into clinical success. PD-1 is a cell
surface inhibitory molecule that is expressed on activated B cells,
T cells and myeloid cells. In 1999, B7-H1 (PD-L1) protein was
discovered (Dong et al., 1999) and was shown to inhibit T cell
responses in vitro (Dong et al., 2002). Subsequent studies showed
that PD-L1 is the ligand for PD-1 and the PD-1/PD-L1 interaction is
a critical mediator of suppressing T cell responses (Freeman et
al., 2000). Thus, the use of therapeutic antibodies to either PD-1
or PD-L1 restores T cell responses (Francisco et al., 2009) and
results in improved clinical outcomes (Brahmer et al., 2010).
[0004] Activated T cells secrete interferon-.gamma. as part of
their immuno-modulatory function, but tumor cells can often coopt
the interferon response by up-regulating PD-L1 and increasing cell
surface expression (Dong et al., 2002). Increased expression of
PD-L1 binds to PD-1 on T cells and suppresses their anti-tumor
(cytolytic) function. Notably, monoclonal antibodies that target
either PD-1 or PD-L1, which effectively disrupt the PD-1/PD-L1
interaction, have been approved by the Food and Drug Administration
(FDA) for a number of indications. However, no small molecule
inhibitors that also impinge on this interaction have been
approved.
[0005] Heat shock proteins (HSPs) are a class of chaperone proteins
that are involved in diverse cellular processes such as elevation
in temperature, external stresses, and nutrient deprivation. Their
basic role as chaperone proteins is to stabilize proteins under
such stresses but also to facilitate the correct folding of client
proteins. There are several members of proteins within HSPs
including HSP27, HSP70 and HSP90. HSP90 is one of the most abundant
family members within cells and is prominently involved in cancer
due to its client proteins which include various oncogenes
including BCR-ABL, BRAF, FLT3, JAK2 and others (Shrestha et al.,
2016). Indeed, HSP90 inhibitors have been used in many pre-clinical
studies to demonstrate how tumor cells are critically dependent on
aberrant pro-survival pathways and how their use results in
inhibition of tumor growth. In this pre-clinical setting, HSP90
inhibitors have shown activity in a variety of cancers including
breast, colorectal, gastro-intestinal, leukemia, lymphomas,
melanoma, multiple myeloma, ovarian, pancreatic, prostate and
renal.
[0006] HSP90 inhibitors have been tested in pre-clinical and early
clinical studies relating to various cancers including breast,
colorectal, gastro-intestinal, leukemia, lymphomas, melanoma,
multiple myeloma, ovarian, pancreatic, prostate and renal. At least
18 HSP90 inhibitors have been investigated in clinical trials,
including BIIB021, IPI-493, MPC-3100, Debio0932, DS-2248, HSP990,
XL888, SNX5422, TAS-116, BIIB028, IPI-504, KW-2478, alvespimycin,
tanespimycin, AT-13387, AUY922, PU-H71 and ganetespib. See reviews
by Bhat et al., J. Med. Chem 2014 57:8718-8728; Neckers and Workman
Clin. Cancer Res. 2012, 18, 64.
[0007] Due to the different chemical properties of each drug and
how they were formulated (oral or intravenous), there is no set
dosage or schedule for administration. However, the common paradigm
applied to other therapeutics such as chemotherapies, or targeted
agents, is that each drug is tested in phase 1 trials at increasing
doses (dose-escalation) to establish safety and eventually
determine the maximum tolerated dose, thus identifying the
recommended phase 2 dose (RP2D). Once reached, the drug is then
tested in phase 2 trials at the identified RP2D on a larger cohort
of patients to determine whether the drug is efficacious.
[0008] Whether HSP90 inhibitors can be effective at a lower dose,
relative to the established RP2D, has not been addressed. Despite
having been tried in various phase 1, 2 and 3 clinical trials, no
HSP90 inhibitor has been approved for clinical use. Given the
number of client proteins dependent on HSP90 and the number of
cellular process these proteins regulate, the question of toxicity
remains a possibility. Thus, the clinical doses used, while having
the propensity to directly affect tumor cells, may also negatively
impact the immune system. This in turn may mitigate any
immune-mediated anti-tumor activity. Efforts to reduce the dose of
HSP90 inhibitors that may negatively impact the immune system while
using sufficient dose to retain HSP90 anti-tumor activity should be
explored and forms the basis of this application.
[0009] Emerging evidence suggests that HSP90 may also affect tumor
immunity. Some non-clinical studies have suggested that high HSP90
inhibitor doses may inhibit various components of the immune system
that may be important for tumor clearance (Bae et al., J. Immunol.
2007 178: 7730; Bae et al., J. Immunol. 2013 190:1360; Tukaj et
al., J. Inflammation 2014 11:10). In addition, many tumor cells
express the checkpoint inhibitor protein death ligand 1 (PD-L1) on
their surface, which can suppress local cytotoxic T cell activity.
For example, PD-L1 expression is found on patient AML cells,
increases with disease progression and during relapse (Salih et
al., Exp. Hematol. 2006 34:888; Chen et al., Cancer Biol. Ther.
2008 7:622; Berthon et al., Cancer Immunol. Immunother 2010
59:1839) and is associated with poorer overall survival (Brodska et
al., Blood 2016 128:5229). PD-L1 cell surface expression on AML
tumor cells may be induced by IFN-.gamma. which is known to be
expressed in the immunologically active tumor microenvironment
(Berthon et al, Cancer Immunol. Immunother 2010 59:1839; Kronig et
al., Eur. J. Hematol. 2013 92:195).
[0010] There is a continuing need for improved treatments and drug
combinations for treating cancer, including the treatment of
cancers that are refractory to current therapies, or those that
have relapsed after treatment. The present invention addresses this
need with the use of HSP90 inhibitors, and in particular in their
combination with inhibitors of the BCL-2 pathway.
SUMMARY OF THE INVENTION
[0011] The disclosure provides compositions and methods related to
the use of an HSP90 inhibitor for treating cancer in a subject,
preferably a human subject, in need of such treatment. The methods
relate generally to the use of low dose HSP90 inhibitors in
pharmaceutical compositions for the treatment of cancer, either
alone, or in combination with other therapies and/or active agents.
The methods further relate generally to the use of HSP90 inhibitors
in combination with BCL-2 pathway inhibitors. As described in more
detail infra, the compositions and methods described here are
based, in part, on the discovery that HSP90 inhibitors are
effective inhibitors of the interferon-.gamma. (IFN-.gamma.) signal
transduction pathway that is activated in diverse types of cancer
cells as a mechanism to evade the immune response. This previously
unreported immunomodulatory activity of HSP90 occurs at relatively
low, non-cytotoxic doses. In addition, the compositions and methods
described here are based, in part, on the discovery of synergistic
activity between HSP90 inhibitors and BCL-2 pathway inhibitors in
cancer.
[0012] The disclosure provides methods for treating cancer in a
subject in need thereof, the methods comprising administering to
the subject an amount of an HSP90 inhibitor and a BCL-2 pathway
inhibitor, optionally wherein the amount of the HSP90 inhibitor is
a sub-therapeutic amount. The disclosure also provides related
pharmaceutical compositions for use in treating cancer, including a
pharmaceutical composition comprising an amount of an HSP90
inhibitor and a BCL-2 pathway inhibitor, and a pharmaceutically
acceptable carrier or excipient, for treating cancer in a subject
in need thereof, optionally wherein the amount of the HSP90
inhibitor is a sub-therapeutic amount; and a pharmaceutical
composition comprising an amount of an HSP90 inhibitor for use in
combination with a second pharmaceutical composition comprising a
BCL-2 pathway inhibitor, for treating cancer in a subject in need
thereof, optionally wherein the amount of the HSP90 inhibitor is a
sub-therapeutic amount.
[0013] In embodiments, the disclosure also provides methods for
treating cancer in a subject in need thereof, the method comprising
determining BCL-2 expression in a biological sample of the cancer
and administering an amount of an HSP90 inhibitor and a BCL-2
pathway inhibitor to the subject having a cancer characterized as
positive for BCL-2 expression based on the expression of BCL-2 in
the biological sample of the cancer, optionally wherein the HSP90
inhibitor and the BCL-2 pathway inhibitor are administered in the
same dosage form or in a different dosage form, optionally wherein
the amount of the HSP90 inhibitor is a sub-therapeutic amount.
[0014] In accordance with the foregoing methods and uses, the
cancer may be further characterized as positive for BCL-2
expression based on the expression of BCL-2 in a biological sample
of the cancer. In embodiments, the cancer characterized as positive
for BCL-2 expression is a cancer in which a biological sample from
the cancer expresses BCL-2 at a level that is at least two-fold
higher compared to the BCL-2 expression in a reference sample, for
example a sample of non-cancerous cells or tissue. In accordance
with the foregoing methods, the BCL-2 expression may be protein
expression or gene expression.
[0015] In accordance with embodiments of the foregoing methods and
uses, the sub-therapeutic amount of the HSP90 inhibitor is less
than 90%, less than 75%, less than 50%, or less than 25% of the
recommended phase 2 dose of the HSP90 inhibitor.
[0016] In accordance with embodiments of the foregoing methods and
uses, the cancer is a hematopoietic or lymphoid cancer selected
from a leukemia, a lymphoma, and a myeloma. In embodiments, the
cancer is a leukemia selected from acute lymphoblastic leukemia
(ALL), acute myelogenous leukemia (AML), chronic lymphocytic
leukemia (CLL), chronic myelogenous leukemia (CML), and acute
monocytic leukemia. In embodiments, the cancer is AML. In
embodiments, the cancer is a lymphoma selected from a Hodgkins and
a Non-Hodgkin's lymphoma. In embodiments, the cancer is a
Non-Hodgkin's B cell lymphoma, preferably selected from a diffuse
large B cell lymphoma (DLBCL), Burkitt lymphoma, lymphoblastic
lymphoma, and mantle cell lymphoma, and most preferably selected
from a diffuse large B cell lymphoma (DLBCL) and a mantle cell
lymphoma. In embodiments, the cancer is a myeloma.
[0017] In accordance with embodiments of the foregoing methods and
uses, the HSP90 inhibitor is selected from a purine-like inhibitor,
a resorcinol derivative, a geldanamycin derivative, a
pyrazolopyridine derivative, a dihydroindazolone derivative, and a
tropane derivative. In embodiments, the HSP90 inhibitor is selected
from WC-0767, AT-13387, tanespimycin, TAS-116, SNX-5422, and
XL-888, and pharmaceutically acceptable salts thereof. In
embodiments, the HSP90 inhibitor is WC-0767 or tanespimycin, and
pharmaceutically acceptable salts thereof. In embodiments, the
HSP90 inhibitor is selected from the group consisting of HSP-990,
CNF-2024, PF0498473, tanespimycin, STA-9090, MPC-3100, CUDC-305,
XL-888, TAS-116, and pharmaceutically acceptable salts thereof. In
embodiments, the HSP90 inhibitor is selected from the group
consisting of tanespimycin, alvespimycin, IPI-504, AUY922,
AT-13387, ganetespib, KW-2478, CNF2024, WC3100, BIIB028, SNX5422,
PU-H71, MPC-0767, and pharmaceutically acceptable salts
thereof.
[0018] In accordance with embodiments of the foregoing methods and
uses, the BCL-2 pathway inhibitor is selected from ABT-737, AT-101
(Gossypol), APG-1252, A1155463, A1210477, navitoclax, obatoclax,
sabutoclax, venetoclax, S 55746, WEHI-539, AMG-176, MIK665, and
S641315. In embodiments, the BCL-2 pathway inhibitor is an
inhibitor of BCL2, BCLXL, or MCL1. In embodiments, the BCL-2
pathway inhibitor is selected from AMG-176, MIK665 and S641315. In
embodiments, the BCL-2 pathway inhibitor is selected from ABT-737,
navitoclax, and venetoclax. In embodiments, the BCL-2 pathway
inhibitor is venetoclax.
[0019] The disclosure also provides methods for treating a BCL-2
expressing hematopoietic or lymphoid cancer in a subject in need
thereof, the methods comprising administering to the subject an
amount of an HSP90 inhibitor and a BCL-2 pathway inhibitor. In
embodiments, the BCL-2 pathway inhibitor is selected from ABT-737,
AT-101 (Gossypol), APG-1252, A1155463, A1210477, navitoclax,
obatoclax, sabutoclax, venetoclax, S 55746, WEHI-539, AMG-176,
MIK665, and S641315. In embodiments, the BCL-2 pathway inhibitor is
venetoclax. In embodiments, the HSP90 inhibitor is selected from a
purine-like inhibitor, a resorcinol derivative, a geldanamycin
derivative, a pyrazolopyridine derivative, a dihydroindazolone
derivative, and a tropane derivative. In embodiments, the HSP90
inhibitor is selected from MPC-0767, AT-13387, tanespimycin,
TAS-116, SNX-5422, and XL-888. In embodiments, the HSP90 inhibitor
is MPC-0767 or tanespimycin.
[0020] In further embodiments of the methods for treating a BCL-2
expressing hematopoietic or lymphoid cancer, the amount of the
HSP90 inhibitor is a sub-therapeutic amount.
[0021] In further embodiments of the methods for treating a BCL-2
expressing hematopoietic or lymphoid cancer, the cancer is a
hematopoietic or lymphoid cancer selected from a leukemia, a
lymphoma, and a myeloma. In embodiments, the cancer is a leukemia
selected from acute lymphoblastic leukemia (ALL), acute myelogenous
leukemia (AML), chronic lymphocytic leukemia (CLL), chronic
myelogenous leukemia (CML), and acute monocytic leukemia. In
embodiments, the cancer is AML. In embodiments, the cancer is a
lymphoma selected from a Hodgkins and a Non-Hodgkin's lymphoma. In
embodiments, the cancer is a Non-Hodgkin's B cell lymphoma,
preferably selected from a diffuse large B cell lymphoma (DLBCL),
Burkitt lymphoma, lymphoblastic lymphoma, and mantle cell lymphoma,
and most preferably selected from a diffuse large B cell lymphoma
(DLBCL) and a mantle cell lymphoma. In embodiments, the cancer is a
myeloma.
[0022] In further embodiments of the methods for treating a BCL-2
expressing hematopoietic or lymphoid cancer, the methods comprise
determining the amount of BCL-2 expressed in a biological sample of
the cancer.
[0023] In accordance with any of the foregoing methods and uses,
the subject may be human.
[0024] In further embodiments, the disclosure provides a
pharmaceutical composition comprising an HSP90 inhibitor, and a
pharmaceutically acceptable carrier or excipient, for use in
treating cancer in a subject in need thereof, wherein the
composition comprises an amount of the HSP90 inhibitor that is less
than 75% of the recommended phase 2 dose of the HSP90
inhibitor.
[0025] In embodiments, the disclosure provides a method for
treating cancer in a subject in need thereof, comprising
administering to the subject a pharmaceutical composition
comprising an amount of an HSP90 inhibitor, and a pharmaceutically
acceptable carrier or excipient, wherein the amount of the HSP90
inhibitor is less than 90% or less than 75% of the recommended
phase 2 dose of the HSP90 inhibitor. In embodiments, the
therapeutically effective amount of an HSP90 inhibitor is the
amount effective to inhibit IFN-.gamma. signaling in cancer cells
of the subject.
[0026] In embodiments of the compositions and methods described
here, the amount of the HSP90 inhibitor is less than 50% or less
than 25% of the recommended phase 2 dose of the HSP90 inhibitor. In
embodiments, the HSP90 inhibitor is selected from the group
consisting of HSP-990, CNF-2024, PF0498473, tanespimycin, STA-9090,
WC-3100, CUDC-305, XL-888, TAS-116, and pharmaceutically acceptable
salts thereof. In embodiments, the HSP90 inhibitor is selected from
the group consisting of tanespimycin, alvespimycin, IPI-504,
AUY922, AT13387, ganetespib, KW-2478, CNF2024, MPC3100, BIIB028,
SNX5422, PU-H71, MPC-0767, and pharmaceutically acceptable salts
thereof.
[0027] In embodiments of the compositions and methods described
here, the pharmaceutical composition comprises a second active
pharmaceutical ingredient (API). In embodiments, second API is
selected from an HDAC inhibitor, an ImiD, an anti-VEGFR antibody, a
DNA methylation inhibitor, a steroid hormone (ant)agonist, a
metabolic enzyme inhibitor, a proteasome inhibitor, an anti-CD20
antibody, an adenosine receptor 2A antagonist, a toll-receptor
(ant(agonist), and an immunostimulatory cytokine. In embodiments,
second API is selected from cisplatin, docetaxel, gemcitabine,
carboplatin, paclitaxel, pemetrexed, etoposide, epirubicin,
doxorubicin, cyclophosphamide, ddAC, everolimus, panobinostat,
exemestane, letrozole, decitabine, esartinib, abemacicib,
merestinib, gefitinib, mocetinostat, azacytidine, etinostat,
motolimod, ibrutinib, lenalidomide, idelalisib, enzalutamide,
olaparib, prednisone, dexamethasone, vinflunine, vorinostat,
galunisertib, bendamustine, oxaliplatin, leucovorin, guadecitabine,
dabrafenib, trametinib, vemurafenib, dacarbazine, apatinib,
pomalidomide, carfilzomib, sorafenib, 5-fluorouracil, CB-839,
CB-1158, GDC-0919, LXH254, AZD4635, AZD9150, PLX3397, LCL161,
PBF-509, bevacizumab, Sym004, ramucirumab, ipilimumab, trastuzumab,
tremelimumab, obinutuzumab, B-701, utomilumab, rituximab,
bevacizumab, interleukin 2, NKTR-214, denenicokin, PEGlnterferon
2A, R07009789, MEDI9447, MK-1248, LY2510924, ARRY-382, MEDI0562,
LAG525; NIS793, Lirilumab, varlilumab, GWN323; JTX-2011;
Galunisertib; TSR-022; BMS-986016, ramucirumab, urelumab,
BMS-986016, REGN3767.
[0028] In embodiments of the compositions and methods described
here, the second API in the composition is selected from the group
consisting of a protein kinase inhibitor, a PD-1/PDL-1 inhibitor, a
checkpoint inhibitor, a platinum based anti-neoplastic agent, a
topoisomerase inhibitor, a nucleoside metabolic inhibitor, an
alkylating agent, an intercalating agent, a tubulin binding agent,
an inhibitor of DNA repair, and combinations thereof. In
embodiments, the second API in the composition is a PD-1/PD-L1
inhibitor. In embodiments, the PD-1/PD-L1 inhibitor is selected
from the group consisting of nivolumab, pembrolizumab,
AMP-514/MEDI-0680, atezolizumab, durvalumab, avelumab, BMS936559,
AMP-224, BGB-A317, SHR-1210, and JTX-4014. In embodiments, the
amount the PD-1/PD-L1 inhibitor is less than 75% of the recommended
phase 2 dose of the PD-1/PD-L1 inhibitor.
[0029] In embodiments of the compositions and methods described
here, the second API in the composition is a CTLA-4 inhibitor. In
embodiments, the CTLA-4 inhibitor is selected from tremlimumab and
ipilimumab. In embodiments, the second API in the composition is a
checkpoint inhibitor. In embodiments, the checkpoint inhibitor is
selected from the group consisting of an anti-CD27 antibody, an
anti-B7-H3 antibody, an anti-KIR antibody, an anti-LAG-3 antibody,
an anti-TIM3 antibody, an anti-OX40 antibody, an anti-4-1BB/CD137
antibody, an anti-CD40 antibody, an anti-TRX518 antibody, an
anti-CD73 antibody, and an anti-GITR antibody. In embodiments, the
checkpoint inhibitor is selected from the group consisting of
varlilumab, MGA217, lirilumab, BMS-986016, urelumab, MEDI-0562,
SEA-CD40, TRX518, and MK-4166. In embodiments, the second API in
the composition is a DNA repair inhibitor selected from the group
consisting of olaparib, rucaparib, niraparib, talazoparib
veliparib, CEP-9722, and CEP-8983.
[0030] In embodiments of the compositions and methods described
here, the cancer is selected from the group consisting of brain
cancer, glioma, sarcoma, breast cancer, lung cancer, non-small-cell
lung cancer, mesothelioma, appendiceal cancer, genitourinary
cancers, renal cell carcinoma, prostate cancer, bladder cancer,
testicular cancer, penile cancer, cervical cancer, ovarian cancer,
head and neck cancer, gastrointestinal cancer, hepatocellular
carcinoma, gallbladder cancer, esophageal cancer, gastric cancer,
colorectal cancer, pancreatic cancer, neuroendocrine tumors,
thyroid tumor, pituitary tumor, adrenal tumor, a T-cell lymphoma,
non-Hodgkin's lymphoma, mantle cell lymphoma, multiple myeloma,
B-cell lymphoma, leukemia, and Hodgkin's lymphoma. In embodiments,
the cancer is selected from the group consisting of melanoma,
Hodgkin's lymphoma, non-small cell lung cancer, bladder cancer,
non-Hodgkin's lymphoma, leukemia, T-cell lymphoma, and renal cell
carcinoma.
[0031] In embodiments of the compositions and methods described
here, the cancer is selected from a cancer that has shown clinical
sensitivity to PD-1/PD-L1 therapies, or a cancer that is utilizing
the PD-1/PD-L1 pathway to evade the host immune system. In
embodiments, the cancer is selected from the group consisting of
melanoma, Hodgkin's lymphoma, non-small cell lung cancer, bladder
cancer, non-Hodgkin's lymphoma, leukemia, T-cell lymphoma, and
renal cell carcinoma.
[0032] In embodiments of the compositions and methods described
here, the pharmaceutical composition comprises an amount of an
HSP90 inhibitor effective to inhibit interferon-.gamma. signal
transduction in cancer cells of the subject.
[0033] In embodiments, the subject is human.
[0034] In embodiments, the pharmaceutical composition is adapted
for oral or buccal administration. In embodiments, the
pharmaceutical composition is adapted for parenteral
administration.
BRIEF DESCRIPTION OF THE FIGURES
[0035] FIG. 1A-D. Validation dose response curves of HSP90
inhibitors tested in the original screen. A) CNF2024; B)
PF04928473; C) tanespimycin; D) HSP-990. SK-MEL-28 cells treated
with IFN-.gamma. and with the indicated HSP90 inhibitor at
different concentrations. Cells were assayed for cell viability
(square, % viability on left axis) and PD-L1 expression (circle,
Z-score right axis) at 48 h after treatment of IFN-.gamma. and/or
HSP90 inhibitors. Quadruplicate wells were used for determination
of cell viability and PD-L1 expression. Average values were
plotted. The inset shows western blot data from SK-MEL-28 cells
treated with vehicle, vehicle+IFN-.gamma., indicated HSP90
inhibitor alone, or IFN-.gamma.+indicated HSP90 inhibitor. Lysates
were probed with anti PD-L1 antibody.
[0036] FIG. 2A-E. Dose response curves of 5 additional HSP90
inhibitors. A) STA-9090, B) CUDC-305, C) MPC-3100, D) XL-888 and E)
TAS-116. SK-MEL-28 cells treated with IFN-.gamma. (50 ng/ml) and
with the indicated HSP90 inhibitor at different concentrations.
Cells were assayed for cell viability (square, % viability on left
axis) and PD-L1 expression (circle, Z-score right axis) at 48 h
after treatment of IFN-.gamma. and/or HSP inhibitors. Quadruplicate
wells were used for determination of cell viability and PD-L1
expression. Average values were plotted.
[0037] FIG. 3. Effect of the HSP90 inhibitor MPC-3100 on the
IFN-.gamma.-induced PD-L1 protein expression. Human HCC-38 cells
treated with IFN-.gamma. alone or co-treated with ruxolitinib (5000
nM) or MPC-3100 (10-5000 nM) for 48 h were analyzed by Western blot
analysis to determine protein expression levels of PD-L1.
Ruxolitinib served as a positive control for inhibiting JAK2.
.beta.-Actin was used as a loading control.
[0038] FIG. 4A-B. Effect of the HSP90 inhibitor MPC-3100 on the
IFN-.gamma.-induced PD-L1 surface signal. A) SK-MEL-28 and HCC-38
(both human lines) were treated with IFN-.gamma. (50 ng/ml),
MPC-3100 (500 nM or 1000 nM) or the combination for 48 h. B) EMT-6
and B16-F10 (both murine cell lines) were also treated with
IFN-.gamma., MPC-3100 or the combination for 24 h. PD-L1 cell
surface signals were analyzed by flow cytometry. Experiments were
performed in duplicate and at least 2 independent times.
[0039] FIG. 5. HCC-38 cells treated with IFN-.gamma. alone or
co-treated with ruxolitinib (5000 nM) or MPC-3100 (10-5000 nM) were
analyzed by Western blot to determine protein expression levels of
HSP70 and pSTAT1. .beta.-Actin was used as a loading control.
[0040] FIG. 6. Effect of HSP90 inhibitor MPC-3100 on
IFN-.gamma.-induced DO protein levels. Human HCC-38 cells treated
with IFN-.gamma. alone or co-treated with ruxolitinib (5000 nM) or
MPC-3100 (100, 1000 or 5000 nM) for 48 h. Lysates were analyzed by
Western blot to determine protein expression levels of DO.
[0041] FIG. 7. Effect of HSP90-inhibitor MPC-3100 on PD-L1 protein
levels. U87 cells were treated with MPC-3100 (500 and 1000 nM) for
24 h and analyzed by Western blotting with the indicated
antibodies. .beta.-Actin was used as a loading control.
[0042] FIG. 8. Stable Disease rate per dose cohort across HSP90
inhibitor Phase I trials.
[0043] FIG. 9. Response rate per dose cohort across HSP90 inhibitor
Phase I trials.
[0044] FIG. 10. Patients with stable disease or clinical response
treated at different % RP2D across all HSP90 drugs that were
analyzed in Phase I trials.
[0045] FIG. 11A-F. Normalized isobolograms at the EC75 of cell
lines representing 4 different indications: Acute Myeloid Leukemia
(MV-4-11), Diffuse Large B Cell Lymphoma (OCI-LY-19), Mantle Cell
Lymphoma (Z138), and Multiple Myeloma (KMS-28) treated with the
combination of venetoclax with each of the following HSP90
inhibitors: A) MPC-0767 B) AT13387 C) Tanespimycin D) SNX5422 E)
XL-888 F) TAS-116, for 72 hours. Cell viability was assessed using
CellTiter-Glo. Each data point is the average of 2 independent
experiments for each cell line. Dashed line denotes `line of
additivity` whereby drug combinations below the line represent
synergistic interactions.
[0046] FIG. 12: Cell lines representing acute myeloid leukemia,
multiple myeloma, diffuse large B cell lymphoma and mantle cell
lymphoma were assayed for basal abundance of BCL-2 (Y axis) and a
high/low threshold is shown by dashed line. Cell line sensitivity
to venetoclax after 72 hour treatment as assessed by CellTiter-Glo
(X axis) is denoted by EC50 values shown in parentheses. Following
drug combination treatment of MPC-0767 and venetoclax,
determination of synergy is shown by bar graphs in black (synergy
observed) or grey (no synergy observed).
[0047] FIG. 13A-D: MPC-0767 blocks IFN-.gamma.-induced PD-L1
expression in AML cells. A) PL-21, B) TUR, C) MOLM-14, D) MV-4-11
cells treated with MPC-0767 (2 .mu.M), IFN-.gamma. (50 ng/ml) or
IFN-.gamma.+MPC-0767 for 24 h before cells were harvested for
assessment of PD-L1 cell surface expression.
DETAILED DESCRIPTION
[0048] Leveraging the immune system has proven to have clinical
efficacy in a number of cancers. By disengaging the inhibitory
interaction of PD-1/PD-L1 between T cells and tumor cells,
checkpoint antibody therapies have changed the paradigm for how a
number of cancers are treated. In embodiments, the disclosure
provides compositions and methods for treating or preventing cancer
in a subject in need thereof, where the cancer is utilizing the
PD-1/PD-L1 pathway to evade the host immune system. The methods
comprise administering to the subject an effective amount of a
HSP90 inhibitor, either alone or in combination with a second
active pharmaceutical agent (API), such as checkpoint inhibitor.
The compositions and methods described here are based, in part, on
the discovery that HSP90 inhibitors are effective to block the
interferon-.gamma. (IFN-.gamma.) signal transduction response of
tumor cells at low, non-cytotoxic doses. Tumor cells utilize
IFN-.gamma. signaling to induce PD-L1, a mechanism for evasion of
the host immune response. Accordingly, in embodiments, the
compositions and methods described here exploit this property of
HSP90 inhibitors to provide new treatments and treatment regimens
for cancer therapy. In embodiments, provided are compositions and
methods related to the use of an HSP90 inhibitor for treating
cancer in a subject, preferably a human subject, in need of such
treatment. The present disclosure generally relates to the use of
HSP90 inhibitors to treat cancers at lower doses than those
previously expected to be effective. For example, at doses that are
less than 90% of the recommended phase 2 dose ("RP2D") of the HSP90
inhibitor, or less than 75%, or less than 50%, or less than 25% of
the recommended phase 2 dose of the HSP90 inhibitor. In the context
of the present disclosure, such doses may also be referred to as
`sub-therapeutic` doses of the HSP90 inhibitor because they are
below the dose expected from prior phase 2 studies to be
therapeutically effective when the HSP90 inhibitor is administered
as monotherapy in the treatment of cancer. In embodiments, a
`sub-therapeutic dose` of an HSP90 inhibitor refers to a dose that
is in the range of 0-25%, 25-50%, 50-75% or 75-90% of the RP2D for
that inhibitor.
[0049] In addition, the disclosure provides compositions and
methods related to the synergistic anti-cancer activity of HSP90
inhibitors generally with inhibitors of the BCL-2 signaling
pathway. As described in more detail below, six different
structural classes of HSP90 inhibitors (defined according to
molecular scaffold) acted synergistically with the BCL-2 inhibitor
venetoclax to inhibit cell viability in haematopoietic and lymphoid
cancers. In addition, the level of BCL-2 expression in the cancer
cells correlated with the anti-cancer activity of the combination.
Accordingly, the disclosure also provides compositions and methods
related to combination therapy with HSP90 inhibitors and BCL-2
pathway inhibitors based on the synergistic anti-cancer activity of
these agents, including methods of identifying cancers likely to
benefit from this combination therapy by assessing the BCL-2
expression of the cancer cells.
HSP90 Inhibitors
[0050] In accordance with the methods and compositions described
here, the HSP90 inhibitor may be any inhibitor of HSP90, for
example, any inhibitor that abrogates the ATPase activity of HSP90.
In embodiments, the HSP90 inhibitor is selected from purine-like
inhibitors, such as MPC-0767, resorcinol derivatives, such as
AT-13387, geldanamycin derivatives, such as tanespimycin,
pyrazolopyridine derivatives, such as TAS-116, dihydroindazolone
derivatives, such as SNX-5422, and tropane derivatives such as
XL-888. Table 1 below shows the chemical structure of a
representative of each of these structural classes of HSP90
inhibitors.
[0051] Accordingly, in embodiments, the HSP90 inhibitor may be
selected from AT-13387, AUY922, B1113028, CNF-2024, CUDC-305,
ganetespib, HSP-990, IPI-504, KW-2478, MPC-0767, WC-3100,
PF0498473, PU-H71, STA-9090, SNX-5422, TAS-116, XL-888,
tanespimycin, alvespimycin, and pharmaceutically acceptable salts
thereof.
TABLE-US-00001 TABLE 1 Representative HSP90 Inhibitors for Each of
Six Chemical Classes HSP90 Inhibitor Class Structure MPC-0767
purine-like ##STR00001## AT-13387 resorcinol derivatives
##STR00002## tanespimycin geldanamycin derivatives ##STR00003##
TAS-116 pyrazolopyridine derivatives ##STR00004## SNX-5422
dihydroindazolone derivatives ##STR00005## XL-888 tropane
derivatives ##STR00006## ##STR00007##
Methods
[0052] The present disclosure provides unique therapeutic
approaches to cancer treatment based upon combination therapy
utilizing an HSP90 inhibitor and a BCL-2 pathway inhibitor. The
compositions and methods relating to combination therapy with HSP90
inhibitors and BCL-2 pathway inhibitors described here exploit the
synergistic anti-cancer activity of these two classes of
therapeutic agents, as described herein. Related methods of
identifying a patient for targeted therapy with a combination of an
HSP90 inhibitor and a BCL-2 pathway inhibitor are also provided. In
embodiments, the methods comprise measuring, determining, or
assaying the expression of BCL-2 in a cancer biopsy or biological
sample or the cancer from the patient. The BCL-2 expression may be
protein expression or gene expression and may, measured according
to routine methods known in the art. For example, gene expression
may be measured by methods comprising a quantitative reverse
transcription polymerase chain reaction (RT-PCR). BCL-2 protein
expression may be measured by methods including, for example,
antibody-based detection methods such as those comprising
immunohistochemistry, immunocytochemistry, enzyme linked
immunosorbent assay (ELISA), and flow cytometry.
[0053] The disclosure also provides methods of treating cancer with
a "low dose" or "sub-therapeutic dose" of an HSP90 inhibitor based
upon the inventors' discovery that such amounts of HSP90 inhibitors
are effective to inhibit the INF.gamma. signaling pathway utilized
by diverse types of cancers to evade the host immune response.
Accordingly, in embodiments, a cancer treated by the "low dose" or
"sub-therapeutic dose" of an HSP90 inhibitor according to the
methods described here is characterized by its utilization of the
PD-1/PD-L1 pathway to evade the host immune system. Non-limiting
examples of such cancers include melanoma, Hodgkin's lymphoma,
non-small cell lung cancer, bladder cancer, non-Hodgkin's lymphoma,
leukemia, T-cell lymphoma, and renal cell carcinoma.
[0054] In addition, the present disclosure provides unique
therapeutic approaches to cancer treatment based upon combination
therapy utilizing a "low dose" or "sub-therapeutic dose" of an
HSP90 inhibitor and at least one additional therapeutic agent. In
embodiments, the combination therapies described herein exploit the
unique immunomodulatory activity of HSP90 inhibitors described
here, which is achieved at a sub-therapeutic dose of the HSP90
inhibitor, and which is expected to provide a synergistic effect
when combined with other therapeutic agents in the treatment of
cancer.
[0055] Both monotherapy and combination therapy methods of treating
cancer with HSP90 inhibitors are contemplated by the present
disclosure. Combination therapies are discussed infra. In the
context of monotherapy, in embodiments the HSP90 inhibitor is
administered at a "low dose" or "sub-therapeutic dose" which is
characterized as an amount of the HSP90 inhibitor that is less than
about 75% of the recommended phase 2 dose of the HSP90 inhibitor.
The terms "low dose" and "sub-therapeutic dose" are used
interchangeably herein in reference to the amount of an HSP90
inhibitor for use in the methods described here.
[0056] In embodiments of the methods described here, the subject in
need of treatment may be one having a cancer that expresses BCL-2,
preferably a cancer that expresses BCL-2 at least two-fold higher
than a reference sample comprising non-cancer cells or tissues.
[0057] In embodiments of the methods described here, the subject in
need of treatment may be one having a cancer that utilizes the
PD-1/PD-L1 pathway to evade the host immune system.
[0058] In embodiments of the methods described here, the subject in
need of treatment may be one having a cancer that is non-responsive
or refractory to, or has relapsed after, treatment with a `standard
of care` or first-line therapeutic agent. In this context, the
terms "non-responsive" and "refractory" are used interchangeably
and refer to the subject's response to therapy as not clinically
adequate, for example to stabilize or reduce the size of one or
more solid tumors, to slow tumor progression, to prevent, reduce or
decrease the incidence of new tumor metastases, or to relieve one
or more symptoms associated with the cancer. A cancer that is
refractory to a particular drug therapy may also be described as a
drug resistant cancer. In a standard therapy for the cancer,
refractory cancer includes disease that in progressing despite
active treatment while "relapsed" cancer includes cancer that
progresses in the absence of any current therapy, but following
successful initial therapy. Accordingly, in embodiments, the
subject is one who has undergone one or more previous regimens of
therapy with one or more `standard of care` therapeutic agents. In
such cases, the subject's cancer may be considered refractory or
relapsed.
[0059] In accordance with the methods described herein, a "subject"
includes a mammal. The mammal can be e.g., any mammal, e.g., a
human, primate, mouse, rat, dog, cat, cow, horse, goat, camel,
sheep or a pig. Preferably, the subject is a human. The term
"patient" refers to a human subject.
[0060] In embodiments, the cancer treated according to the methods
described here is selected from the group consisting of brain
cancer, glioma, sarcoma, breast cancer, lung cancer, non-small-cell
lung cancer, mesothelioma, appendiceal cancer, genitourinary
cancers, renal cell carcinoma, prostate cancer, bladder cancer,
testicular cancer, penile cancer, cervical cancer, ovarian cancer,
head and neck cancer, gastrointestinal cancer, hepatocellular
carcinoma, gallbladder cancer, esophageal cancer, gastric cancer,
colorectal cancer, pancreatic cancer, neuroendocrine tumors,
thyroid tumor, pituitary tumor, adrenal tumor, a T-cell lymphoma,
non-Hodgkin's lymphoma, mantle cell lymphoma, multiple myeloma,
B-cell lymphoma, leukemia, and Hodgkin's lymphoma.
[0061] In embodiments, the cancer is selected from a cancer that
expresses BCL-2, preferably one the expresses BCL-2 at least
two-fold higher compared to a reference non-cancerous tissue.
[0062] In embodiments, the cancer is a hematopoietic or lymphoid
cancer selected from a leukemia, a lymphoma, and a myeloma. In
embodiments, the cancer is a leukemia selected from acute
lymphoblastic leukemia (ALL), acute myelogenous leukemia (AML),
chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia
(CML), and acute monocytic leukemia. In embodiments, the cancer is
AML. In embodiments, the cancer is a lymphoma selected from a
Hodgkins and a Non-Hodgkin's lymphoma. In embodiments, the cancer
is a Non-Hodgkin's B cell lymphoma, preferably selected from a
diffuse large B cell lymphoma (DLBCL), Burkitt lymphoma,
lymphoblastic lymphoma, and mantle cell lymphoma, and most
preferably selected from a diffuse large B cell lymphoma (DLBCL)
and a mantle cell lymphoma. In embodiments, the cancer is a
myeloma.
[0063] As used herein, "combination therapy" or "co-therapy"
includes the administration of a therapeutically effective amount
of an HSP90 inhibitor as part of a treatment regimen intended to
provide the beneficial effect from the co-action of the an HSP90
inhibitor and at least one additional "active pharmaceutical
ingredient" ("API"). "Combination therapy" is not intended to
encompass the administration of two or more therapeutic compounds
as part of separate monotherapy regimens that incidentally and
arbitrarily result in a beneficial effect that was not intended or
predicted.
[0064] Preferably, the administration of a composition comprising
an HSP90 inhibitor in combination with one or more additional APIs,
such as a BCL-2 pathway inhibitor, as discussed herein provides a
synergistic response in the subject being treated. In this context,
the term "synergistic" refers to the efficacy of the combination
being more effective than the additive effects of either single
therapy alone. The synergistic effect of a combination therapy
according to the disclosure can permit the use of lower dosages,
for example a low dose of the HSP90 inhibitor as described here,
and/or less frequent administration of at least one agent in the
combination compared to its dose and/or frequency outside of the
combination. Additional beneficial effects of the combination can
be manifested in the avoidance or reduction of adverse or unwanted
side effects associated with the use of either therapy in the
combination alone (also referred to as monotherapy).
[0065] In the context of combination therapy, administration of the
HSP90 inhibitor composition may be simultaneous with or sequential
to the administration of the one or more additional active agents,
such as the BCL-2 pathway inhibitor. In another embodiment,
administration of the different components of a combination therapy
may be at different frequencies.
[0066] The additional API(s) can be formulated for
co-administration with the HSP90 inhibitor composition in a single
dosage form. The additional API(s) can also be administered
separately from the dosage form that comprises the HSP90 inhibitor.
When the additional active agent is administered separately from
HSP90 inhibitor, it can be by the same or a different route of
administration, and/or at the same or different time.
[0067] In embodiments, the at least one additional API may be a
BCL-2 pathway inhibitor, a protein kinase inhibitor, a PD-1/PD-L1
inhibitor, a checkpoint inhibitor, a platinum based anti-neoplastic
agent, a topoisomerase inhibitor, a nucleoside metabolic inhibitor,
an alkylating agent, an intercalating agent, a tubulin binding
agent, an inhibitor of DNA repair, and combinations thereof. In
embodiments, the at least one additional API is a BCL-2 pathway
inhibitor or a PD-1/PD-L1 inhibitor. In embodiments, a
sub-therapeutic amount of the HSP90 inhibitor is administered with
the BCL-2 pathway inhibitor or the PD-1/PD-L1 inhibitor.
[0068] In embodiments, the at least one additional API is a
PD-1/PD-L1 inhibitor. In embodiments, the PD-1/PD-L1 inhibitor is
selected from AMP-224, AMP-514/MEDI-0680, atezolizumab (MPDL3280A),
avelumab (MSB0010718C), BGB-A317, BMS936559, cemiplimab (REGN2810),
durvalumab (MEDI-4736), JTX-4014, nivolumab (BMS-936558),
pembrolizumab (Keytruda, MK-3475), GLS-010, and SHR-1210.
[0069] In embodiments, the at least one additional API is a BCL-2
pathway inhibitor. In embodiments, the BCL-2 pathway inhibitor is
selected from ABT-737, AT-101 (Gossypol), APG-1252, A1155463,
A1210477, navitoclax, obatoclax, sabutoclax, venetoclax, S 55746,
and WEHI-539. In embodiments, the BCL-2 pathway inhibitor is an
inhibitor of BCL2, BCLXL, or MCL1. In embodiments, the BCL-2
pathway inhibitor is selected from AMG-176, MIK665 and S641315. In
embodiments, the BCL-2 pathway inhibitor is selected from ABT-737,
navitoclax, and venetoclax. In embodiments, the BCL-2 pathway
inhibitor is venetoclax.
[0070] In embodiments, the at least one additional API is a CTLA-4
inhibitor. In embodiments, the CTLA-4 inhibitor is selected from
tremlimumab and ipilimumab.
[0071] In embodiments, the at least one additional API is a check
point inhibitor. Treatment with these compounds works by targeting
molecules that serve as checks and balances on immune responses. By
blocking these inhibitory molecules or, alternatively, activating
stimulatory molecules, these treatments are designed to unleash or
enhance pre-existing anti-cancer immune responses. In embodiments,
the check point inhibitor may be selected from an antibody such as
an anti-CD27 antibody, an anti-B7-H3 antibody, an anti-KIR
antibody, an anti-LAG-3 antibody, an anti-4-1BB/CD137 antibody, an
anti-GITR antibody (e.g., TRX518, MK-4166), pembrolizumab
(Keytruda.TM., a PD-1 antibody), MPDL3280A (a PD-L1 antibody),
varlilumab (CDX-1127, an anti-CD27 antibody), MGA217 (an antibody
that targets B7-H3), lirilumab (a KIR antibody), BMS-986016 (a
LAG-3 antibody), urelumab (a 4-1BB/CD137 antibody), an anti-TIM3
antibody, MEDI-0562 (a OX40 antibody), SEA-CD40 (an anti-CD40
antibody), tremelimumab (anti-CTLA4 antibody), an anti-OX40
antibody, and an anti-CD73 antibody. In embodiments, the checkpoint
inhibitor is selected from a small molecule inhibitor of CD73 (as
described, for example, in Cancer Immunol Res 2016; 4 (11
Suppl):Abstract nr PR10). In embodiments, the checkpoint inhibitor
is selected from varlilumab, MGA217, lirilumab, BMS-986016,
urelumab, MEDI-0562, SEA-CD40, TRX518, or MK-4166. In embodiments,
the at least one additional API is a DNA repair inhibitor. In
embodiments, the DNA repair inhibitor is selected from the group
consisting of olaparib, rucaparib, niraparib, talazoparib
veliparib, CEP-9722, and CEP-8983.
[0072] In embodiments, the at least one additional API is a DNA
repair inhibitor selected from olaparib, rucaparib, niraparib,
talazoparib veliparib, CEP-9722, and CEP-8983.
[0073] In embodiments, the at least one additional API is a VEGF
inhibitor. In embodiments, the VEGF inhibitor is selected from
sunitinib, pazopanib, bevacizumab, sorafenib, cabozantinb, and
axitinib.
[0074] In embodiments, at least one additional API is selected from
ddAC, panobinostat, exemestane, letrozole, esartinib, merestinib,
mocetinostat, etinostat, motolimod, ibrutinib, lenalidomide,
idelalisib, enzalutamide, prednisone, dexamethasone, vinflunine,
vorinostat, galunisertib, bendamustine, oxaliplatin, leucovorin,
guadecitabine, trametinib, vemurafenib, dacarbazine, apatinib,
pomalidomide, carfilzomib, sorafenib, 5-fluorouracil, CB-839,
CB-1158, GDC-0919, LXH254, AZD4635, AZD9150, PLX3397, LCL161,
PBF-509, Sym004, trastuzumab, obinutuzumab, B-701, utomilumab,
rituximab, NKTR-214, PEGlnterferon 2A, R07009789, MEDI9447,
MK-1248, LY2510924, ARRY-382, MEDI0562, LAG525, NIS793, GWN323,
JTX-2011, TSR-022, and REGN3767.
[0075] In embodiments, the at least one additional API is directed
towards targeted therapy, wherein the treatment targets the
cancer's specific genes, proteins, or the tissue environment that
contributes to cancer growth and survival. This type of treatment
blocks the growth and spread of cancer cells while limiting damage
to healthy cells. In embodiments, the at least one additional API
is directed towards anti-angiogenesis therapy, wherein the
treatment focuses on stopping angiogenesis, which is the process of
making new blood vessels. Because a tumor needs the nutrients
delivered by blood vessels to grow and spread, the goal of
anti-angiogenesis therapies is to "starve" the tumor. One
anti-angiogenic drug, bevacizumab (Avastin), has been shown to slow
tumor growth for people with metastatic renal carcinoma.
Bevacizumab combined with interferon slows tumor growth and
spread.
[0076] In embodiments, the at least one additional API is directed
towards immunotherapy, also called biologic therapy, which is
designed to boost the body's natural defenses to fight cancer. It
uses materials made either by the body or in a laboratory to
improve, target, or restore immune system function. For example,
interleukin-2 (IL-2) is a drug that has been used to treat kidney
cancer as well as AM0010, and interleukin-15. They are cellular
hormones called cytokines produced by white blood cells and are
important in immune system function, including the destruction of
tumor cells. Alpha-interferon is another type of immunotherapy used
to treat kidney cancer that has spread. Interferon appears to
change the proteins on the surface of cancer cells and slow their
growth. Many combination therapies of IL-2 and alpha-interferon for
patients with advanced kidney cancer combined with chemotherapy are
more effective than IL-2 or interferon alone.
[0077] In embodiments, the at least one additional API is a cancer
vaccine, designed to elicit an immune response against
tumor-specific or tumor-associated antigens, encouraging the immune
system to attack cancer cells bearing these antigens. In
embodiments, the cancer vaccine is AGS-003, DCVax, NY-ESO-1 or a
personalized vaccine derived from patient's cancer cells.
[0078] In embodiments, the at least one additional API is an
immunostimulant, such as a recombinant protein, used to activate
the immune system to attack cancer cells. In embodiments, the
immunostimulant is denenicokin (recombinant IL-21).
[0079] In embodiments, the at least one additional API is a small
molecule that modulates the immune system to encourage the
elimination of cancer cells. In embodiments, the small molecule is
epacadostat or navoximod (both IDO inhibitors), or PLX3397 (an
inhibitor of CSF-1R).
[0080] In embodiments, the at least one additional API is selected
from taxol, vincristine, doxorubicin, idarubicin, temsirolimus,
carboplatin, ofatumumab, rituximab, and combinations thereof.
[0081] In embodiments, the at least one additional API is selected
from chlorambucil, ifosphamide, doxorubicin, mesalazine,
thalidomide, lenalidomide, temsirolimus, everolimus, fludarabine,
cytarabine, mitoxantrone, fostamatinib, paclitaxel, docetaxel,
ofatumumab, rituximab, dexamethasone, prednisone, CAL-101,
ibritumomab, tositumomab, bortezomib, pentostatin, endostatin, or a
combination thereof.
[0082] In embodiments, the at least one additional API may be the
patient's own immune cells which have been removed from a patient,
genetically modified or treated with chemicals to enhance their
activity, and then re-introduced into the patient with the goal of
improving the immune system's anti-cancer response.
[0083] "Combination therapy" also embraces the administration of
HSP90 inhibitors as described herein in further combination with
non-drug therapies (e.g., surgery or radiation treatment). Where
the combination therapy further comprises a non-drug treatment, the
non-drug treatment may be conducted at any suitable time so long as
a beneficial effect from the co-action of the combination of the
therapeutic compounds and non-drug treatment is achieved. For
example, in appropriate cases, the beneficial effect is still
achieved when the non-drug treatment is temporally removed from the
administration of the therapeutic compounds, perhaps by days or
even weeks.
[0084] The non-drug treatment can be selected from chemotherapy,
radiation therapy, hormonal therapy, anti-estrogen therapy, gene
therapy, surgery (e.g. radical nephrectomy, partial nephrectomy,
laparoscopic and robotic surgery), radiofrequency ablation, and
cryoablation. For example, a non-drug therapy is the removal of an
ovary (e.g., to reduce the level of estrogen in the body),
thoracentesis (e.g., to remove fluid from the chest), paracentesis
(e.g., to remove fluid from the abdomen), surgery to remove or
shrink angiomyolipomas, lung transplantation (and optionally with
an antibiotic to prevent infection due to transplantation), or
oxygen therapy (e.g., through a nasal cannula containing two small
plastic tubes or prongs that are placed in both nostrils, through a
face mask that fits over the nose and mouth, or through a small
tube inserted into the windpipe through the front of the neck, also
called transtracheal oxygen therapy).
[0085] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is selected from a non-small
cell lung cancer (NSCLC) and the at least one additional API is
selected from cisplatin/docetaxel, bevacizumab, gemcitabine,
carboplatin, (nab-) paclitaxel, pemetrexed, etoposide, Sym004
(anti-EGFR), gefitinib, mocetinostat, PLX3397, etinostat, AZD4635
(A2aR antagonist), tremelimumab, ipilimumab, and PBF-509 (A2AR
antagonist).
[0086] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is selected from a solid
tumor and the at least one additional API is selected from
ramucirumab, abemacicib, merestinib, R07009789 (anti-CD40),
MEDI9447 (anti-CD73), MK-1248 (anti-GITR), olaparib, cediranib,
SFU, leucovorin, oxaliplatin, ibrutinib, LY2510924 (CXCR4
antagonist), ARRY-382 (CSFR1i), MEDI0562 (anti-OX40), LAG525
(anti-LAG3), NIS793, lirilumab (anti-KIR), NKTR-214 (selective
IL-2); varlilumab (anti-CD27), IL-21 (denenicokin); GWN323
(anti-GITR); JTX-2011 (anti-ICOS), galunisertib; TSR-022
(anti-TIM3); BMS-986016 (anti-LAG3), REGN3767 (anti-LAG3); GDC-0919
(IDO inh), CB-1158 (Arginase inh), and AZD4635 (A2aR
antagonist).
[0087] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is selected from
mesothelioma, colorectal cancer, urothelial cancer, gastric cancer,
and liver cancer and the at least one additional API is selected
from tremelimumab and ipilimumab.
[0088] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is selected from breast
cancer and the at least one additional API is selected from
nab-paclitaxel, epirubicin, doxorubicin, cyclophosphamide, ddAC,
everolimus, panobinostat, LCL161 (IAP inh), anti-estrogen therapy
(exemestane), letrozole, decitabine, and trastuzumab.
[0089] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is an ALK positive cancer,
such as an ALK positive lung cancer, including some forms of NSCLC,
and the at least one additional API is selected from esartinib.
[0090] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is a urothelial cancer and
the at least one additional API is selected from
gemcitabine/carboplatin, docetaxel, paclitaxel, vinflunine, B-701
(anti-FGFR3) and vorinostat.
[0091] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is an ovarian cancer and the
at least one additional API is selected from a multi-epitope
anti-folate vaccine, motolimod, carboplatin, and paclitaxel.
[0092] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is a renal cell cancer (RCC)
and the at least one additional API is selected from etinostat,
bevacizumab, IL-2, vorinostat, and CB-839 (glutaminase inh).
[0093] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is pancreatic cancer and the
at least one additional API is galunisertib.
[0094] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is gastric cancer and the at
least one additional API is selected from trastuzumab,
capecitabine, cisplatin, margetuximab, and apatinib.
[0095] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is liver cancer and the at
least one additional API is selected from apatinib, and
sorafenib.
[0096] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is myelodysplastic syndrome
(MDS) and the at least one additional API is selected from
azacytidine, tremelimumab, and etinostat.
[0097] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is chronic lymphocytic
leukemia (CLL) and the at least one additional API is selected from
obinutuzumab, ibrutinib, lenalidomide, rituximab, and
bendamustine.
[0098] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is metastatic colorectal or
prostate cancer and the at least one additional API is selected
from sipuleucel-T, enzalutamide, olaparib, docetaxel, prednisone,
and dexamethasone.
[0099] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is diffuse large B cell
lymphoma (DLBCL) and the at least one additional API is selected
from KTE-19, AZD9150 (STAT3 inh), utomilumab, rituximab,
azacytidine, bendamustine, gemcitabine, oxaliplatin, and
R-CHOP.
[0100] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is a glioblastoma and the at
least one additional API is selected from urelumab (anti-4-1BB) and
BMS 986016 (Anti-LAG-3).
[0101] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is multiple myeloma (MM) and
the at least one additional API is selected from lenalidomide,
dexamethasone, carfilzomib, daratumumab (anti-CD38), and
pomalidomide.
[0102] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is a gastrointestinal or
thoracic cancer and the at least one additional API is ramucirumab
(anti-VEGFR2).
[0103] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is a head and neck cancer and
the at least one additional API is selected from
cisplatin/carboplatin, SFU, cetuximab, and SD-101 (anti-TLR9).
[0104] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount, the cancer is acute myeloid leukemia (AML)
and the at least one additional API is selected from azacytidine,
crenolanib, cytarabine, daunorubicin, etoposide, gilteritinib,
guadecitabine, idarubicin, midostaurin, mitoxantrone, quizartinib,
sorafenib, tandutinib, and venetoclax. In embodiments, the at least
one additional API is selected from crenolanib, cytarabine,
daunorubicin, gilteritinib, sorafenib, and venetoclax. In
embodiments, the at least one additional API is venetoclax. In
embodiments, the at least one additional API is selected from an
anthracycline, such as daunorubicin, doxorubicin, epirubicin,
mitoxantrone, and idarubicin; cytarabine; a tyrosine kinase
inhibitor (TKI) such as midostaurin, sorefenib, crenolanib,
quizartinib, tandutinib, gilteritinib, lestaurtinib, dovitinib,
pacritinib, and XL999; etoposide, fludarabine, G-CSF, azacytidine,
decitabine, venetoclax, ABT-737, navitoclax, obatoclax, sabutoclax,
S 55746, AT-101 (Gossypol), and APG-1252, and combinations of any
of the foregoing. In embodiments, the at least one additional API
is selected from arsenic trioxide (trisenox), cerubidine
(Daunorubicin Hydrochloride), clafen (Cyclophosphamide),
cyclophosphamide, cytarabine (tarabine PFS), cytosar-U
(Cytarabine), cytoxan (Cyclophosphamide), daunorubicin
hydrochloride (rubidomycin), doxorubicin hydrochloride, enasidenib
mesylate, idamycin (idarubicin hydrochloride), idarubicin
hydrochloride idhifa (Enasidenib Mesylate), midostaurin (Rydapt),
mitoxantrone hydrochloride, neosar (Cyclophosphamide), thioguanine
(Tabloid), vincristine sulfate (vincasar PFS), azacytidine, and
decitabine, and combinations of any of the foregoing. In
embodiments, the at least one additional API is a PD-1/PD-L1
inhibitor selected from the group consisting of AMP-224,
AMP-514/MEDI-0680, atezolizumab (MPDL3280A), avelumab
(MSB0010718C), BGB-A317, BMS936559, cemiplimab (REGN2810),
durvalumab (MEDI-4736), JTX-4014, nivolumab (BMS-936558),
pembrolizumab (Keytruda, MK-3475), GLS-010 and SHR-1210. In
embodiments, the at least one additional API is a BCL-2 pathway
inhibitor selected from the group consisting of ABT-737, AT-101
(Gossypol), APG-1252, A1155463, A1210477, navitoclax, obatoclax,
sabutoclax, venetoclax, S 55746, and WEHI-539. In embodiments, the
BCL-2 pathway inhibitor is an inhibitor of BCL2, BCLXL, or MCL1. In
embodiments, the BCL-2 pathway inhibitor is selected from AMG-176,
MIK665 and S641315. In embodiments, the BCL-2 pathway inhibitor is
selected from ABT-737, navitoclax, and venetoclax.
[0105] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is melanoma and the at least
one additional API is selected from dabrafenib, trametinib, PLX3397
(CSF-R1 inh), vemurafenib, IFN.alpha.2B, dacarbazine, carboplatin,
paclitaxel, and SD-101 (anti-TLR9).
[0106] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount and the cancer is non-Hodgkins lymphoma and
the at least one additional API is selected from JCAR014.
[0107] In embodiments, the amount of the HSP90 inhibitor is a
sub-therapeutic amount, the cancer is a renal cancer, and the
anti-cancer agent may be selected from a VEGF inhibitor such as
sunitinib, pazopanib, bevacizumab, sorafenib, cabozantinb and
axitinib or an mTOR inhibitor such as everolimus or
temsirolimus.
[0108] Generally, where two different APIs are administered, one is
considered the primary agent and the other is an adjunct to the
primary agent. In embodiments of the methods described here, the
HSP90 inhibitor may be either the primary or adjunct agent in the
therapeutic regimen. In embodiments, the adjunct agent is
administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45
minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48
hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5
weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with,
or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45
minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48
hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5
weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of
the primary agent in the therapeutic regimen.
[0109] In the context of the methods described herein, the amount
of an HSP90 inhibitor administered to the subject is a
therapeutically effective amount. The term "therapeutically
effective amount" refers to an amount sufficient to treat,
ameliorate a symptom of, reduce the severity of, or reduce the
duration of the disease or disorder being treated or enhance or
improve the therapeutic effect of another therapy, or sufficient to
exhibit a detectable therapeutic effect in the subject. In
embodiments, the therapeutically effective amount of an HSP90
inhibitor in the methods described here is a dose that would be
considered `sub-therapeutic` or below the dose expected to be
effective based on phase 2 clinical studies of the HSP90 inhibitor.
In embodiments, a sub-therapeutic dose of an HSP90 inhibitor is a
dose that is less than 75% of the recommended phase 2 dose of the
HSP90 inhibitor, or less than 50%, or less than 25% of the
recommended phase 2 dose of the HSP90 inhibitor. In one embodiment,
the therapeutically effective amount of an HSP90 inhibitor is the
amount effective to inhibit IFN-.gamma. signaling in cancer cells
of the subject. In embodiments, the methods comprise administering
a sub-therapeutic dose of an HSP90 inhibitor, alone or in
combination with one or more additional APIs.
[0110] As used herein, "treatment", "treating" or "treat" describes
the management and care of a patient for the purpose of combating a
disease, condition, or disorder and includes the administration of
an HSP90 inhibitor, alone as monotherapy (for example, utilizing
low dose HSP90) or in combination with at least one additional API
as described here, to alleviate the symptoms or complications of a
disease, condition or disorder, or to eliminate the disease,
condition or disorder.
[0111] In embodiments, including both monotherapy with an HSP90
inhibitor and combination therapies with one or more additional
APIs, the administration of an HSP90 inhibitor leads to the
elimination of a symptom or complication of the cancer being
treated, however elimination of the cancer is not required. In one
embodiment, the severity of the symptom is decreased. In the
context of cancer, such symptoms may include clinical markers of
severity or progression including the degree to which a tumor
secretes growth factors, degrades the extracellular matrix, becomes
vascularized, loses adhesion to juxtaposed tissues, or
metastasizes, as well as the number of metastases and reduction in
tumor size and/or volume.
[0112] Treating cancer according to the methods described herein
can result in a reduction in size of a tumor. A reduction in size
of a tumor may also be referred to as "tumor regression".
Preferably, after treatment, tumor size is reduced by 5% or greater
relative to its size prior to treatment; more preferably, tumor
size is reduced by 10% or greater; more preferably, reduced by 20%
or greater; more preferably, reduced by 30% or greater; more
preferably, reduced by 40% or greater; even more preferably,
reduced by 50% or greater; and most preferably, reduced by greater
than 75% or greater. Size of a tumor may be measured by any
reproducible means of measurement. The size of a tumor may be
measured as a diameter of the tumor.
[0113] Treating cancer according to the methods described herein
can result in a reduction in tumor volume. Preferably, after
treatment, tumor volume is reduced by 5% or greater relative to its
size prior to treatment; more preferably, tumor volume is reduced
by 10% or greater; more preferably, reduced by 20% or greater; more
preferably, reduced by 30% or greater; more preferably, reduced by
40% or greater; even more preferably, reduced by 50% or greater;
and most preferably, reduced by greater than 75% or greater. Tumor
volume may be measured by any reproducible means of
measurement.
[0114] Treating cancer according to the methods described herein
can result in a decrease in number of tumors. Preferably, after
treatment, tumor number is reduced by 5% or greater relative to
number prior to treatment; more preferably, tumor number is reduced
by 10% or greater; more preferably, reduced by 20% or greater; more
preferably, reduced by 30% or greater; more preferably, reduced by
40% or greater; even more preferably, reduced by 50% or greater;
and most preferably, reduced by greater than 75%. Number of tumors
may be measured by any reproducible means of measurement. The
number of tumors may be measured by counting tumors visible to the
naked eye or at a specified magnification. Preferably, the
specified magnification is 2.times., 3.times., 4.times., 5.times.,
10.times., or 50.times.. For hematologic cancers, the count may be
the number of cells related to the cancer (e.g., lymphoma or
leukemia cells) in a sample of blood.
[0115] Treating cancer according to the methods described herein
can result in a decrease in the number of metastatic lesions in
other tissues or organs distant from the primary tumor site.
Preferably, after treatment, the number of metastatic lesions is
reduced by 5% or greater relative to the number prior to treatment;
more preferably, the number of metastatic lesions is reduced by 10%
or greater; more preferably, reduced by 20% or greater; more
preferably, reduced by 30% or greater; more preferably, reduced by
40% or greater; even more preferably, reduced by 50% or greater;
and most preferably, reduced by greater than 75%. The number of
metastatic lesions may be measured by any reproducible means of
measurement. The number of metastatic lesions may be measured by
counting metastatic lesions visible to the naked eye or at a
specified magnification. Preferably, the specified magnification is
2.times., 3.times., 4.times., 5.times., 10.times., or
50.times..
[0116] Treating cancer according to the methods described herein
can result in an increase in average survival time of a population
of treated subjects in comparison to a population receiving carrier
alone. Preferably, the average survival time is increased by more
than 30 days; more preferably, by more than 60 days; more
preferably, by more than 90 days; and most preferably, by more than
120 days. An increase in average survival time of a population may
be measured by any reproducible means. An increase in average
survival time of a population may be measured, for example, by
calculating for a population the average length of survival
following initiation of treatment. An increase in average survival
time of a population may also be measured, for example, by
calculating for a population the average length of survival
following completion of a first round of treatment.
[0117] Treating cancer according to the methods described herein
can result in an increase in average survival time of a population
of treated subjects in comparison to a population of untreated
subjects. Preferably, the average survival time is increased by
more than 30 days; more preferably, by more than 60 days; more
preferably, by more than 90 days; and most preferably, by more than
120 days. An increase in average survival time of a population may
be measured by any reproducible means. An increase in average
survival time of a population may be measured, for example, by
calculating for a population the average length of survival
following initiation of treatment. An increase in average survival
time of a population may also be measured, for example, by
calculating for a population the average length of survival
following completion of a first round of treatment.
[0118] Treating cancer according to the methods described herein
can result in an increase in average survival time of a population
of treated subjects in comparison to a population receiving
monotherapy with a drug that is not an HSP90 inhibitor. Preferably,
the average survival time is increased by more than 30 days; more
preferably, by more than 60 days; more preferably, by more than 90
days; and most preferably, by more than 120 days. An increase in
average survival time of a population may be measured by any
reproducible means. An increase in average survival time of a
population may be measured, for example, by calculating for a
population the average length of survival following initiation of
treatment. An increase in average survival time of a population may
also be measured, for example, by calculating for a population the
average length of survival following completion of a first round of
treatment.
[0119] Treating cancer according to the methods described herein
can result in a decrease in the mortality rate of a population of
treated subjects in comparison to a population receiving carrier
alone. Treating a disorder, disease or condition according to the
methods described herein can result in a decrease in the mortality
rate of a population of treated subjects in comparison to an
untreated population. Treating a disorder, disease or condition
according to the methods described herein can result in a decrease
in the mortality rate of a population of treated subjects in
comparison to a population receiving monotherapy with a drug that
is not an HSP90 inhibitor. Preferably, the mortality rate is
decreased by more than 2%; more preferably, by more than 5%; more
preferably, by more than 10%; and most preferably, by more than
25%. A decrease in the mortality rate of a population of treated
subjects may be measured by any reproducible means. A decrease in
the mortality rate of a population may be measured, for example, by
calculating for a population the average number of disease-related
deaths per unit time following initiation of treatment. A decrease
in the mortality rate of a population may also be measured, for
example, by calculating for a population the average number of
disease-related deaths per unit time following completion of a
first round of treatment.
[0120] Treating cancer according to the methods described herein
can result in a decrease in tumor growth rate. Preferably, after
treatment, tumor growth rate is reduced by at least 5% relative to
number prior to treatment; more preferably, tumor growth rate is
reduced by at least 10%; more preferably, reduced by at least 20%;
more preferably, reduced by at least 30%; more preferably, reduced
by at least 40%; more preferably, reduced by at least 50%; even
more preferably, reduced by at least 50%; and most preferably,
reduced by at least 75%. Tumor growth rate may be measured by any
reproducible means of measurement. Tumor growth rate can be
measured according to a change in tumor diameter per unit time. In
one embodiment, after treatment the tumor growth rate may be about
zero and is determined to maintain the same size, e.g., the tumor
has stopped growing.
[0121] Treating cancer according to the methods described herein
can result in a decrease in tumor regrowth. Preferably, after
treatment, tumor regrowth is less than 5%; more preferably, tumor
regrowth is less than 10%; more preferably, less than 20%; more
preferably, less than 30%; more preferably, less than 40%; more
preferably, less than 50%; even more preferably, less than 50%; and
most preferably, less than 75%. Tumor regrowth may be measured by
any reproducible means of measurement. Tumor regrowth is measured,
for example, by measuring an increase in the diameter of a tumor
after a prior tumor shrinkage that followed treatment. A decrease
in tumor regrowth is indicated by failure of tumors to reoccur
after treatment has stopped.
[0122] Pharmaceutical Compositions and Formulations
[0123] The present disclosure provides pharmaceutical compositions
comprising an amount of an HSP90 inhibitor, either alone or in
combination with at least one additional API, such as a BCL-2
pathway inhibitor. In embodiments, the amount of the HSP90
inhibitor is less than 75% of the recommended phase 2 dose of the
HSP90 inhibitor. In embodiments, the amount of the HSP90 inhibitor
is less than 75% of the recommended phase 2 dose of the HSP90
inhibitor. In accordance with any of the embodiments described
here, the pharmaceutical composition may be adapted for oral,
buccal, or parenteral administration. In embodiments, the
pharmaceutical composition may be adapted for pulmonary
administration, for example by inhalation. In embodiments, the
pharmaceutical composition is adapted for oral administration. In
embodiments, the pharmaceutical composition is adapted for
parenteral administration.
[0124] In embodiments, the HSP90 inhibitor is combined with at
least one additional API in a single dosage form. In embodiments,
the at least one additional API is selected from an agent described
supra in connection with methods of treatment using combination
therapy.
[0125] A "pharmaceutical composition" is a formulation containing
the compounds described herein in a pharmaceutically acceptable
form suitable for administration to a subject. As used herein, the
phrase "pharmaceutically acceptable" refers to those compounds,
materials, compositions, carriers, and/or dosage forms which are,
within the scope of sound medical judgment, suitable for use in
contact with the tissues of human beings and animals without
excessive toxicity, irritation, allergic response, or other problem
or complication, commensurate with a reasonable benefit/risk
ratio.
[0126] "Pharmaceutically acceptable excipient" means an excipient
that is useful in preparing a pharmaceutical composition that is
generally safe, non-toxic and neither biologically nor otherwise
undesirable, and includes excipient that is acceptable for
veterinary use as well as human pharmaceutical use. Examples of
pharmaceutically acceptable excipients include, without limitation,
sterile liquids, water, buffered saline, ethanol, polyol (for
example, glycerol, propylene glycol, liquid polyethylene glycol and
the like), oils, detergents, suspending agents, carbohydrates
(e.g., glucose, lactose, sucrose or dextran), antioxidants (e.g.,
ascorbic acid or glutathione), chelating agents, low molecular
weight proteins, or suitable mixtures thereof.
[0127] A pharmaceutical composition can be provided in bulk or in
dosage unit form. It is especially advantageous to formulate
pharmaceutical compositions in dosage unit form for ease of
administration and uniformity of dosage. The term "dosage unit
form" as used herein refers to physically discrete units suited as
unitary dosages for the subject to be treated; each unit containing
a predetermined quantity of active compound calculated to produce
the desired therapeutic effect in association with the required
pharmaceutical carrier. The specification for the dosage unit forms
of the disclosure are dictated by and directly dependent on the
unique characteristics of the active compound and the particular
therapeutic effect to be achieved. A dosage unit form can be an
ampoule, a vial, a suppository, a dragee, a tablet, a capsule, an
IV bag, or a single pump on an aerosol inhaler.
[0128] In therapeutic applications, the dosages vary depending on
the agent, the age, weight, and clinical condition of the recipient
patient, and the experience and judgment of the clinician or
practitioner administering the therapy, among other factors
affecting the selected dosage. Generally, the dose should be a
therapeutically effective amount. Dosages can be provided in
mg/kg/day units of measurement (which dose may be adjusted for the
patient's weight in kg, body surface area in m.sup.2, and age in
years). An effective amount of a pharmaceutical composition is that
which provides an objectively identifiable improvement as noted by
the clinician or other qualified observer. For example, alleviating
a symptom of a disorder, disease or condition. As used herein, the
term "dosage effective manner" refers to amount of a pharmaceutical
composition to produce the desired biological effect in a subject
or cell.
[0129] For example, the dosage unit form can comprise 1 nanogram to
2 milligrams, or 0.1 milligrams to 2 grams; or from 10 milligrams
to 1 gram, or from 50 milligrams to 500 milligrams or from 1
microgram to 20 milligrams; or from 1 microgram to 10 milligrams;
or from 0.1 milligrams to 2 milligrams.
[0130] The pharmaceutical compositions can take any suitable form
(e.g, liquids, aerosols, solutions, inhalants, mists, sprays; or
solids, powders, ointments, pastes, creams, lotions, gels, patches
and the like) for administration by any desired route (e.g,
pulmonary, inhalation, intranasal, oral, buccal, sublingual,
parenteral, subcutaneous, intravenous, intramuscular,
intraperitoneal, intrapleural, intrathecal, transdermal,
transmucosal, rectal, and the like). For example, a pharmaceutical
composition of the disclosure may be in the form of an aqueous
solution or powder for aerosol administration by inhalation or
insufflation (either through the mouth or the nose), in the form of
a tablet or capsule for oral administration; in the form of a
sterile aqueous solution or dispersion suitable for administration
by either direct injection or by addition to sterile infusion
fluids for intravenous infusion; or in the form of a lotion, cream,
foam, patch, suspension, solution, or suppository for transdermal
or transmucosal administration.
[0131] A pharmaceutical composition can be in the form of an orally
acceptable dosage form including, but not limited to, capsules,
tablets, buccal forms, troches, lozenges, and oral liquids in the
form of emulsions, aqueous suspensions, dispersions or solutions.
Capsules may contain mixtures of a compound of the present
disclosure with inert fillers and/or diluents such as the
pharmaceutically acceptable starches (e.g., corn, potato or tapioca
starch), sugars, artificial sweetening agents, powdered celluloses,
such as crystalline and microcrystalline celluloses, flours,
gelatins, gums, etc. In the case of tablets for oral use, carriers
which are commonly used include lactose and corn starch.
Lubricating agents, such as magnesium stearate, can also be added.
For oral administration in a capsule form, useful diluents include
lactose and dried corn starch. When aqueous suspensions and/or
emulsions are administered orally, the compound of the present
disclosure may be suspended or dissolved in an oily phase is
combined with emulsifying and/or suspending agents. If desired,
certain sweetening and/or flavoring and/or coloring agents may be
added.
[0132] A pharmaceutical composition can be in the form of a tablet.
The tablet can comprise a unit dosage of a compound of the present
disclosure together with an inert diluent or carrier such as a
sugar or sugar alcohol, for example lactose, sucrose, sorbitol or
mannitol. The tablet can further comprise a non-sugar derived
diluent such as sodium carbonate, calcium phosphate, calcium
carbonate, or a cellulose or derivative thereof such as methyl
cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and
starches such as corn starch. The tablet can further comprise
binding and granulating agents such as polyvinylpyrrolidone,
disintegrants (e.g. swellable crosslinked polymers such as
crosslinked carboxymethylcellulose), lubricating agents (e.g.
stearates), preservatives (e.g. parabens), antioxidants (e.g. BHT),
buffering agents (for example phosphate or citrate buffers), and
effervescent agents such as citrate/bicarbonate mixtures.
[0133] The tablet can be a coated tablet. The coating can be a
protective film coating (e.g. a wax or varnish) or a coating
designed to control the release of the active agent, for example a
delayed release (release of the active after a predetermined lag
time following ingestion) or release at a particular location in
the gastrointestinal tract. The latter can be achieved, for
example, using enteric film coatings such as those sold under the
brand name Eudragit.RTM..
[0134] Tablet formulations may be made by conventional compression,
wet granulation or dry granulation methods and utilize
pharmaceutically acceptable diluents, binding agents, lubricants,
disintegrants, surface modifying agents (including surfactants),
suspending or stabilizing agents, including, but not limited to,
magnesium stearate, stearic acid, talc, sodium lauryl sulfate,
microcrystalline cellulose, carboxymethylcellulose calcium,
polyvinylpyrrolidone, gelatin, alginic acid, acacia gum, xanthan
gum, sodium citrate, complex silicates, calcium carbonate, glycine,
dextrin, sucrose, sorbitol, dicalcium phosphate, calcium sulfate,
lactose, kaolin, mannitol, sodium chloride, talc, dry starches and
powdered sugar. Preferred surface modifying agents include nonionic
and anionic surface modifying agents. Representative examples of
surface modifying agents include, but are not limited to, poloxamer
188, benzalkonium chloride, calcium stearate, cetostearyl alcohol,
cetomacrogol emulsifying wax, sorbitan esters, colloidal silicon
dioxide, phosphates, sodium dodecylsulfate, magnesium aluminum
silicate, and triethanolamine.
[0135] A pharmaceutical composition can be in the form of a hard or
soft gelatin capsule. In accordance with this formulation, the
compound of the present disclosure may be in a solid, semi-solid,
or liquid form.
[0136] A pharmaceutical composition can be in the form of a sterile
aqueous solution or dispersion suitable for parenteral
administration. The term parenteral as used herein includes
subcutaneous, intracutaneous, intravenous, intramuscular,
intra-articular, intraarterial, intrasynovial, intrasternal,
intrathecal, intralesional and intracranial injection or infusion
techniques.
[0137] A pharmaceutical composition can be in the form of a sterile
aqueous solution or dispersion suitable for administration by
either direct injection or by addition to sterile infusion fluids
for intravenous infusion, and comprises a solvent or dispersion
medium containing, water, ethanol, a polyol (e.g., glycerol,
propylene glycol and liquid polyethylene glycol), suitable mixtures
thereof, or one or more vegetable oils. Solutions or suspensions of
the compound of the present disclosure as a free base or
pharmacologically acceptable salt can be prepared in water suitably
mixed with a surfactant. Examples of suitable surfactants are given
below. Dispersions can also be prepared, for example, in glycerol,
liquid polyethylene glycols and mixtures of the same in oils.
[0138] The pharmaceutical compositions for use in the methods of
the present disclosure can further comprise one or more additives
in addition to any carrier or diluent (such as lactose or mannitol)
that is present in the formulation. The one or more additives can
comprise or consist of one or more surfactants. Surfactants
typically have one or more long aliphatic chains such as fatty
acids which enables them to insert directly into the lipid
structures of cells to enhance drug penetration and absorption. An
empirical parameter commonly used to characterize the relative
hydrophilicity and hydrophobicity of surfactants is the
hydrophilic-lipophilic balance ("HLB" value). Surfactants with
lower HLB values are more hydrophobic, and have greater solubility
in oils, while surfactants with higher HLB values are more
hydrophilic, and have greater solubility in aqueous solutions.
Thus, hydrophilic surfactants are generally considered to be those
compounds having an HLB value greater than about 10, and
hydrophobic surfactants are generally those having an HLB value
less than about 10. However, these HLB values are merely a guide
since for many surfactants, the HLB values can differ by as much as
about 8 HLB units, depending upon the empirical method chosen to
determine the HLB value.
[0139] Among the surfactants for use in the compositions of the
disclosure are polyethylene glycol (PEG)-fatty acids and PEG-fatty
acid mono and diesters, PEG glycerol esters, alcohol-oil
transesterification products, polyglyceryl fatty acids, propylene
glycol fatty acid esters, sterol and sterol derivatives,
polyethylene glycol sorbitan fatty acid esters, polyethylene glycol
alkyl ethers, sugar and its derivatives, polyethylene glycol alkyl
phenols, polyoxyethylene-polyoxypropylene (POE-POP) block
copolymers, sorbitan fatty acid esters, ionic surfactants,
fat-soluble vitamins and their salts, water-soluble vitamins and
their amphiphilic derivatives, amino acids and their salts, and
organic acids and their esters and anhydrides.
[0140] The present disclosure also provides packaging and kits
comprising pharmaceutical compositions for use in the methods of
the present disclosure. The kit can comprise one or more containers
selected from the group consisting of a bottle, a vial, an ampoule,
a blister pack, and a syringe. The kit can further include one or
more of instructions for use in treating and/or preventing a
disease, condition or disorder of the present disclosure, one or
more syringes, one or more applicators, or a sterile solution
suitable for reconstituting a pharmaceutical composition of the
present disclosure.
[0141] All percentages and ratios used herein, unless otherwise
indicated, are by weight. Other features and advantages of the
present disclosure are apparent from the different examples. The
provided examples illustrate different components and methodology
useful in practicing the present disclosure. The examples do not
limit the claimed disclosure. Based on the present disclosure the
skilled artisan can identify and employ other components and
methodology useful for practicing the present disclosure.
[0142] In embodiments, the disclosure provides the following.
[0143] 1. A pharmaceutical composition comprising an HSP90
inhibitor, and a pharmaceutically acceptable carrier or excipient,
for use in treating cancer in a subject in need thereof, wherein
the composition comprises an amount of the HSP90 inhibitor that is
less than 75% of the recommended phase 2 dose of the HSP90
inhibitor.
[0144] 2. A method for treating cancer in a subject in need
thereof, comprising administering to the subject a pharmaceutical
composition comprising an amount of an HSP90 inhibitor, and a
pharmaceutically acceptable carrier or excipient, wherein the
amount of the HSP90 inhibitor is less than 75% of the recommended
phase 2 dose of the HSP90 inhibitor.
[0145] 3. The method of claim 1 or 2, wherein the amount of the
HSP90 inhibitor is less than 50% or less than 25% of the
recommended phase 2 dose of the HSP90 inhibitor.
[0146] 4. The method of any one of claims 1-3, wherein the HSP90
inhibitor is selected from the group consisting of HSP-990,
CNF-2024, PF0498473, tanespimycin, STA-9090, MPC-3100, CUDC-305,
XL-888, TAS-116, and pharmaceutically acceptable salts thereof.
[0147] 5. The method of any one of claims 1-3, wherein the HSP90
inhibitor is selected from the group consisting of tanespimycin,
alvespimycin, IPI-504, AUY922, AT13387, ganetespib, KW-2478,
CNF2024, MPC3100, BIIB028, SNX5422, PU-H71, MPC-0767, and
pharmaceutically acceptable salts thereof.
[0148] 6. The method of any one of claims 1-5, wherein the
pharmaceutical composition comprises a second active pharmaceutical
ingredient (API).
[0149] 7. The method of claim 6, wherein the second API is selected
from an HDAC inhibitor, an ImiD, an anti-VEGFR antibody, a DNA
methylation inhibitor, a steroid hormone (ant)agonist, a metabolic
enzyme inhibitor, a proteasome inhibitor, an anti-CD20 antibody, an
adenosine receptor 2A antagonist, a toll-receptor (ant(agonist), an
immunostimulatory cytokine, and combinations thereof.
[0150] 8. The method of claim 6, wherein the second API is selected
from cisplatin, docotaxel, gemcitabine, carboplatin, paclitaxel,
pemetrexed, etoposide, epirubicin, doxorubicin, cyclophosphamide,
ddAC, everolimus, panobinostat, exemestane, letrozole, decitabine,
esartinib, abemacicib, merestinib, gefitinib, mocetinostat,
azacytidine, etinostat, motolimod, ibrutinib, lenalidomide,
idelalisib, enzalutamide, olaparib, prednisone, dexamethasone,
vinflunine, vorinostat, galunisertib, bendamustine, oxaliplatin,
leucovorin, guadecitabine, dabrafenib, trametinib, vemurafenib,
dacarbazine, apatinib, pomalidomide, carfilzomib, sorafenib,
5-fluorouracil, CB-839, CB-1158, GDC-0919, LXH254, AZD4635,
AZD9150, PLX3397, LCL161, PBF-509, bevacizumab, Sym004,
ramucirumab, ipilimumab, trastuzumab, tremelimumab, obinutuzumab,
B-701, utomilumab, rituximab, bevacizumab, interleukin 2, NKTR-214,
denenicokin, PEGInterferon 2A, R07009789, MEDI9447, MK-1248,
LY2510924, ARRY-382, MEDI0562, LAG525; NIS793, Lirilumab,
varlilumab, GWN323; JTX-2011; Galunisertib; TSR-022; BMS-986016,
ramucirumab, urelumab, BMS-986016, and REGN3767.
[0151] 9. The method of claim 6, wherein the second API in the
composition is selected from the group consisting of a protein
kinase inhibitor, a PD-1/PD-L1 inhibitor, a checkpoint inhibitor, a
platinum based anti-neoplastic agent, a topoisomerase inhibitor, a
nucleoside metabolic inhibitor, an alkylating agent, an
intercalating agent, a tubulin binding agent, an inhibitor of DNA
repair, and combinations thereof.
[0152] 10. The method of claim 9, wherein the second API in the
composition is a PD-1/PD-L1 inhibitor.
[0153] 11. The method of claim 10, wherein the PD-1/PD-L1 inhibitor
is selected from the group consisting of nivolumab, pembrolizumab,
AMP-514/MEDI-0680, atezolizumab, durvalumab, avelumab, BMS936559,
AMP-224, BGB-A317, SHR-1210, and JTX-4014.
[0154] 12. The method of claim 10 or 11, wherein the amount the
PD-1/PD-L1 inhibitor is less than 75% of the recommended phase 2
dose of the PD-1/PD-L1 inhibitor.
[0155] 13. The method of claim 6, wherein the second API in the
composition is a CTLA-4 inhibitor.
[0156] 14. The method of claim 13, wherein the CTLA-4 inhibitor is
selected from tremlimumab and ipilimumab.
[0157] 15. The method of claim 9, wherein the second API in the
composition is a checkpoint inhibitor.
[0158] 16. The method of claim 15, wherein the checkpoint inhibitor
is selected from the group consisting of an anti-CD27 antibody, an
anti-B7-H3 antibody, an anti-KIR antibody, an anti-LAG-3 antibody,
an anti-TIM3 antibody, an anti-OX40 antibody, an anti-4-1BB/CD137
antibody, an anti-CD40 antibody, an anti-TRX518 antibody, an
anti-CD73 antibody, and an anti-GITR antibody.
[0159] 17. The method of claim 15, wherein the checkpoint inhibitor
is selected from the group consisting of varlilumab, MGA217,
lirilumab, BMS-986016, urelumab, MEDI-0562, SEA-CD40, TRX518, and
MK-4166.
[0160] 18. The method of claim 9, wherein the second API in the
composition is a DNA repair inhibitor selected from the group
consisting of olaparib, rucaparib, niraparib, talazoparib
veliparib, CEP-9722, and CEP-8983.
[0161] 19. The method of any one of claims 1-18, wherein the cancer
is selected from the group consisting of brain cancer, glioma,
sarcoma, breast cancer, lung cancer, non-small-cell lung cancer,
mesothelioma, appendiceal cancer, genitourinary cancers, renal cell
carcinoma, prostate cancer, bladder cancer, testicular cancer,
penile cancer, cervical cancer, ovarian cancer, head and neck
cancer, gastrointestinal cancer, hepatocellular carcinoma,
gallbladder cancer, esophageal cancer, gastric cancer, colorectal
cancer, pancreatic cancer, neuroendocrine tumors, thyroid tumor,
pituitary tumor, adrenal tumor, a T-cell lymphoma, non-Hodgkin's
lymphoma, mantle cell lymphoma, multiple myeloma, B-cell lymphoma,
leukemia, and Hodgkin's lymphoma.
[0162] 20. The method of claim 19, wherein the cancer is selected
from the group consisting of melanoma, Hodgkin's lymphoma,
non-small cell lung cancer, bladder cancer, non-Hodgkin's lymphoma,
leukemia, T-cell lymphoma, and renal cell carcinoma.
[0163] 21. The method of claim 10, 11, or 12, wherein the cancer is
selected from the group consisting of melanoma, Hodgkin's lymphoma,
non-small cell lung cancer, bladder cancer, non-Hodgkin's lymphoma,
leukemia, T-cell lymphoma, and renal cell carcinoma.
[0164] 22. The method of any one of claims 1-21, wherein the
pharmaceutical composition comprises an amount of an HSP90
inhibitor effective to inhibit interferon-.gamma. signal
transduction in cancer cells of the subject.
[0165] 23. The method of any one of claims 1-22, wherein the
subject is human.
[0166] 24. The method of any one of claims 1-23, wherein the
pharmaceutical composition is adapted for oral or buccal
administration.
[0167] 25. The method of any one of claims 1-23, wherein the
pharmaceutical composition is adapted for parenteral
administration.
EXAMPLES
Example 1--a Screen to Identify Small Molecule Inhibitors of PD-L1
Cell Surface Expression
[0168] SK-MEL-28 cells (melanoma cell line; ATCC.RTM. HTB-72.TM.)
were used to conduct a high-content screen to identify small
molecules that reduce interferon-gamma (IFN-.gamma.) (Shenandoah
Biotechnology) induced PD-L1 expression.
[0169] Cells were expanded in MEM (Corning) containing 10% FBS
(Sigma) and 2 mM L-Glutamine. Frozen stocks of cells were prepared
for direct use in the high-throughput screening (HTS) assay. Cells
were harvested, pelleted and then resuspended in 95% FBS & 5%
DMSO at a concentration 1.times.10.sup.7 cells/ml. One ml aliquots
were rate frozen to -80.degree. C. at a rate of 1 degree per
minute. These stocks were then transferred to vapor phase liquid
nitrogen for long term storage.
[0170] For screening, frozen cell vials were quickly thawed with
continuous agitation in a at 37.degree. C. water bath then
re-suspended in the assay media at room temperature and centrifuged
at 1,000 rpm for 5 minutes. The resulting pellet was re-suspended
in appropriate volume and counted using an automated cell counter
and diluted accordingly.
[0171] Using a manually curated library of approximately 2500
individual drugs (2000 approved drugs, 500 unapproved drugs), drugs
from the source plate were transferred to the destination plates
(384-well assay plates--Corning #3712) using an ECHO 550 liquid
handler (Labcyte). SK-MEL-28 cells (1750 cells per well in 304, of
media), treated with IFN-g (50 ng/ml final) were added to these
pre-formatted plates using a Multidrop.TM. Combi reagent dispenser
(Thermofisher). The final concentration of the drugs was 40, 480
and 5000 nM. The outer 2 columns on the left of the plate and the
right of the plate (columns 1, 2, 23, 24) served as positive and
negative controls and were treated with and without IFN-.gamma.,
respectively. Plates were centrifuged at 800 rpm for one minute
before being incubated in a 5% CO.sub.2 humidified incubator at
37.degree. C.
[0172] 44 h later, cell viability was determined using
alamarBlue.TM. cell viability reagent (Thermofisher), according to
the manufacturers protocol. Briefly, 3 .mu.L of alamarBlue.TM.
reagent was added to each well and plates were incubated for a
further 4 h. The plates were read on a Victor .sup.3V plate reader
(Perkin Elmer). Viability for each drug was expressed as a
percentage by comparing drug-treated cells and untreated control
cells (Set to 100%).
[0173] After assessment for viability, plates were then prepared
for immunostaining. Briefly, 16% paraformaldehyde (Electron
Microscopy Sciences) was added to each well to a final
concentration of 4%, for 10 minutes at room temperature. Following
this time, the fix was aspirated and cells were washed 3 times with
PBS. Cells were blocked using blocking solution (1% bovine serum
albumin in PBS) for 30 minutes. After this time, the blocking
reagent was aspirated and immunostaining reagent added. The
immunostaining reagent consisted of a biotin conjugated-PD-L1
antibody (eBioscience) diluted in blocking buffer. Plates were
sealed and kept at 4C overnight. The following day, plates were
washed 3 times with PBS followed by addition of a
streptavidin-luciferase fusion protein (Invivogen) diluted in
blocking buffer, for 1 h at room temperature. After this step, the
plates were washed 3 times with PBS. After the final wash, PBS was
aspirated and 30 uL of QUANTI-Luc.TM. (Invivogen) was added per
well. Plates were immediately read on a plate reader (Victor 3V
plate reader, 0.1 s) and luciferase activity measured.
[0174] The raw PD-L1 intensity (the luciferase readout)
corresponding to drug screen plates was normalized to blank plates
containing only IFN-.gamma.-treated wells, and the resulting ratio
was used for the calculation of Z-scores.
[0175] The Z-scores were calculated for each drug taking into
account the blank-corrected PD-L1 intensity and the control
+IFN-.gamma.-treated wells in each drug plate, according to the
following equation:
Z score = I Drug - mean ( I + IFN ) sd ( I + IFN ) ##EQU00001##
where I.sup.Drug is the blank-corrected PD-L1 intensity for each
drug. Therefore, Z score values will be negative for those drugs
that decrease PD-L1 levels with respect to the IFN-.gamma.-treated
wells.
[0176] In order to compare the PD-L1 effect of different drug doses
tested across different plates, we defined a relative Z score, that
takes into account the drug Z score and the distribution of Z
scores corresponding to the IFN-.gamma. untreated control
wells.
relative Z score = Z score Drug mean ( Z score - IFN )
##EQU00002##
[0177] The relative Z score is a measure of drug PD-L1 level effect
with respect to the baseline PD-L1 signal.
[0178] Once we computed the relative Z scores for all drugs and all
concentrations tested in the screen, we applied three filtering
criteria to narrow down drug list and select high-confidence hits,
as follows: [0179] #1 Select drugs with a relative Z
score<=-1+sd (Z score.sup.-IFN-.gamma.) and a viability percent
>75% for the three concentrations tested. This rendered 10 drugs
at 40 nM, 27 drugs at 480 nM and 196 drugs at 5000 nM (out of 4126
total drugs). [0180] #2 Select drugs at 5000 nM that showed a hit
percent >50%. This resulted in 110 drugs at 5000 nM (out of 196
obtained in #1). [0181] #3 Select drugs that showed a
dose-responsive relative Z score. This resulted in 47 drugs.
[0182] To summarize the results from the primary screen, 47 drugs
reduced IFN-.gamma.-induced PD-L1 expression. Notably, these
included small molecule inhibitors of JAK2, which have previously
been shown to abrogate PD-L1 expression (Green et al., 2010).
[0183] Of the 47 drug identified in the primary screen, we focused
our efforts on the HSP90 inhibitor class of drugs since there were
4 individual small molecule inhibitors targeting HSP90 in our
library of -2500 drugs (HSP-990, CNF-2024, PF0498473 and
tanespimycin), and all 4 were identified as hits in the primary
screen.
[0184] These 4 HSP90 inhibitors were validated using the same
experimental protocol employed above (using the SK-MEL-28
cells+/-IFN-.gamma. in 384 well plates), with the only exception to
the original assay design was that each drug was now tested in a
10-point dose response assay where the concentration of drug tested
was 9.8-5000 nM (2 fold serial dilution). Cells were seeded and
treated in quadruplicate. Viability was assessed after 48 h of
treatment of four HSP 90 inhibitors and prior to harvesting the
cells for testing PD-L1 signal, as described above.
[0185] In this secondary assay all 4 HSP90 drugs (HSP-990,
CNF-2024, PF0498473 and tanespimycin) were validated by confirming
that IFN-.gamma.-induced PD-L1 signal was blocked (FIG. 1A-D).
[0186] Thus, these findings serve to demonstrate that we have
uncovered a mechanistic link between inhibition of HSP90 and
abrogating IFN-.gamma.-induced PD-L1 expression.
[0187] Previous studies noted that inhibition of HSP90 reduced JAK2
levels and suggested JAK2 could be a client protein of HSP90
(Marubayashi et al., 2010)(Proia et al., 2011). However, to date,
no direct link of HSP90 inhibition and PD-L1 expression has been
made. The results reported here suggest that HSP90 inhibitors can
have a therapeutic effect by modulating the immune response toward
instances where the PD-1/PD-L1 axis is usurped to evade immune
surveillance. This mechanism can be exploited, as described herein,
to provide a new approach to cancer treatment using low dose HSP90
inhibitors, alone or in combination with other active agents.
[0188] When comparing the concentrations at which the HSP90
inhibitors exerted their immuno-modulatory effects on PD-L1 versus
their anti-proliferative effects, it was evident that the effects
of HSP90 inhibitors on inhibition of IFN-.gamma.-induced PD-L1
expression occurred at concentrations much lower than those
required for anti-proliferative activity (see FIG. 1A-D).
[0189] We therefore sought to extend the preliminary findings from
our HTS screen by testing additional HSP90 inhibitors not present
in our library of -2500 drugs. STA-9090, MPC-3100, CUDC-305, XL-888
and TAS-116, were tested for their ability to inhibit
IFN-.gamma.-induced PD-L1 expression. As shown in FIG. 2A-E, the 5
additionally tested HSP90 inhibitors showed the same trend--the
concentration required to inhibit IFN-.gamma.-induced PD-L1
expression (red line) was far lower than the concentration required
to affect cell proliferation since viability remained higher than
80% at the highest concentration tested (1000 nM).
[0190] To confirm that our findings are applicable to other cell
types, an additional human breast cancer cell line HCC-38 was used.
HCC-38 cells were treated with IFN-.gamma. (50 ng/ml) and
co-treated with MPC-3100 at 10, 100, 1000 and 5000 nM for 48 h. As
shown in FIG. 3, IFN-.gamma.-induced PD-L1 expression was reduced
by MPC-3100 at 100 nM and was completely abrogated at 1000 nM.
[0191] Considering the maximum concentration (Cmax, geometric mean)
of MPC-3100 in the patients' plasma dosed at the recommended phase
2 dose is -13 .mu.M, the concentrations at which MPC-3100 elicited
immuno-modulatory activity toward PD-L1 is at least 10 fold
lower.
[0192] To verify that the activity of MPC-3100 on abrogating
IFN-.gamma.-induced PD-L1 expression as detected by western
blotting, which measures total intracellular protein, resulted in
decreased cell surface expression, flow cytometry was performed.
Two human cell lines were used (SK-MEL-28 and HCC-38 cells) and two
murine cell lines were used (B16-F10, a melanoma cell line and
EMT-6, a breast cancer cell line). All cells lines were treated
with IFN-.gamma. (50 ng/nl) and co-treated with MPC-3100 at the
indicated concentrations for 48 h. As shown in FIG. 4A-B, all cell
lines tested showed that MPC-3100 effectively inhibited
IFN-.gamma.-induced cell surface expression of PD-L1.
[0193] Since IFN-.gamma. stimulates the expression of its target
genes, such as PD-L1, which is mediated via JAK/STAT-1 signaling,
the human HCC-38 cell line was used to probe the mechanism for
IFN-.gamma.-induced PD-L1 expression. As shown in FIG. 5, MPC-3100
blocks IFN-induced p-STAT1, suggesting this is the mechanism for
how MPC-3100 blocks IFN-.gamma.-induced PD-L1 expression. Again,
the concentrations at which this occurred was at least 10 fold
lower than the Cmax at the recommended Phase 2 dose of the
drug.
[0194] Consistent with the findings above, the drug ruxolitinb, (a
JAK2 inhibitor) also blocked IFN-induced PD-L1 expression through
its inhibitory activity toward JAK2, as measured by inhibition of
p-STAT.
[0195] Numerous studies have shown that exposing cells to HSP90
inhibitors results in a paradoxical increase in HSP70 expression,
both in vitro and in vivo. As shown in FIG. 5, HCC-38 cells treated
with MPC-3100 also show an increase in HSP70 expression.
[0196] Based on these findings, we tested whether indoleamine
2,3-dioxygenase (IDO), another IFN-.gamma. induced protein which is
implicated in immune suppression (Curti et al., 2009), can be
blocked by a HSP90 inhibitor. As expected, HCC-38 cells treated
with IFN-.gamma. alone induced the expression of IDO. However, when
HCC-38 cells were co-treated with IFN-.gamma. and MPC-3100 (1000 nM
or 5000 nM) IDO expression was inhibited (FIG. 6).
[0197] Collectively, these findings establish that HSP90
inhibitors, through blocking STAT-1 signaling, can effectively
block IFN-.gamma.-induced PD-L1 and IDO expression. Although HSP90
inhibitors display anti-proliferative activity, the effects on
abrogating IFN-.gamma.-induced PD-L1 expression occurs at
concentrations that are not cytotoxic. This finding was
demonstrated in different cell types as well as in different
species, suggesting this phenomenon should be broadly translatable.
Moreover, these findings show that MPC-3100 has biological activity
in blocking IFN-.gamma.-mediated signaling at concentrations that
do not reduce cell viability and are also at sub-cytotoxic
concentrations.
[0198] In addition to PD-L1 expression being regulated by
IFN-.gamma. in an inducible manner, several studies have shown that
genetic perturbations can lead to constitutive expression of PD-L1
(Parsa et al., 2007). Indeed, glioma lines with mutations in PTEN
demonstrate hyperactivation of Akt, resulting in constitutive PD-L1
expression. The levels of PD-L1 are significantly higher as
compared with glioma cell lines with wild-type PTEN (Parsa et al.,
2007).
[0199] To test whether HSP90 inhibitors are effective in reducing
constitutive expression of PD-L1, U87 cells (human glioma) were
treated with MPC-3100 at 500 or 1000 nM for 24 h. Lysates were then
prepared and queried for protein expression using western blot
analysis. As shown in FIG. 7, U87 cells do not have basal
activation of JAK-STAT-1 signaling as evidenced by lack of
detectable p-STAT. However, since these cells harbor mutant PTEN,
there is hyperactivation of AKT signaling as evidenced by pAkt.
Moreover, as expected from the Parsa et al. study, there is high
basal expression of PD-L1. However, in U87 cells treated with
WC-3100, there is a reduction in pAkt and attenuation of PD-L1.
These data demonstrate that under conditions where an oncogene
induces expression of PD-L1, MPC-3100 is able to block inhibit
PD-L1 expression. Furthermore, inhibition of PD-L1 expression
occurs at concentrations .about.10 fold lower than the therapeutic
dose.
[0200] To look at whether low doses of HSP90 inhibitors exhibited
higher or similar clinical efficacy than the RP2D, we performed a
meta-analysis on Phase I clinical trials that included 12 different
inhibitors across 20 different trials (Saif et al., 2014; Bauer et
al., 2013; Siegal et al., 2011; Pacey et al., 2011; Yong et al.,
2016; Reddy et al., 2013; Isambert et al., 2015; Doi et al., 2014;
Padmanabhan et al., 2010; Mahadevan et al., 2012; Maddocks et al.,
2016; Lancet et al., 2010; Hong et al., 2013; Solit et al., 2007;
Nowakowski et al., 2006; Goldman et al., 2013; Kummar et al., 2010;
Cho et al., 2011; Wagner et al., 2013; LAM Therapeutics,
confidential). Only patients having stable disease for at least 4
treatment cycles or a clinical response were analyzed. The doses
tested in the different trials were normalized with respect to the
recommended Phase 2 dose per drug and indication and the percentage
of dose (% RP2D) was calculated for each dose cohort. Patients with
stable disease or clinical response across all the trials were then
grouped according to the % RP2D at which they were treated, in
order to calculate the percent of stable disease (% Stable Disease)
and percent of response (% Response) per dose cohort across all the
trials.
[0201] The results show that more occurrences of stable disease
(FIG. 8) and clinical response (FIG. 9) are achieved at lower doses
of HSP90 inhibitor than the recommended Phase 2 dose. This is when
the HSP90 inhibitor is 0-25% of the RP2D, when the HSP90 inhibitor
is 25-50% of the RP2D and when the HSP90 inhibitor is 50-75% of the
RP2D. This observation reaches statistical significance (Fisher
Test P-value=0.005) when all doses lower that 75% of the RP2D are
combined. A similar trend is observed for either solid or
hematopoietic and lymphoid tumors (FIG. 10), since patients with
stable disease or clinical response are observed at lower doses
regardless of their tumor type. Moreover our analysis suggests that
the observed "low dose clinical efficacy" is a common feature of
the HSP90 drug class, since all HSP90 inhibitors that were analyzed
show efficacy at doses lower than the RP2D (FIG. 10). This suggests
that the premise of this application should hold true for any of
the HSP90 inhibitors used in the clinic.
[0202] These results support the hypothesis that HSP90 inhibitors
have higher clinical efficacy when administered at lower doses thus
supporting the immuno-oncology role for this drug class in cancer
treatment.
[0203] Thus, collectively, the data presented for HSP90 inhibitors
showing clinical activity at doses of 0-25%, 25-50%, 50-75% or
75-90% of the RP2D, could be translated into the clinic either as
single agent or could reduce the dose required for combined
therapies with checkpoint antibodies targeting the PD-1/PD-L1 axis.
This includes antibodies that target PD-1 such as Nivolumab,
Pembrolizumab, AMP-224, BGB-317 (China), SHR-1210, JTX-4014,
AMP-514/MEDI-0680, GLS-010, or antibodies that target PD-L1 such as
Atezolizumab, Durvalumab, Avelumab and BMS936559 (MDX-1105). Given
the synergy between CTLA-4 and PD-1/PD-L1 immuno-therapies, lower
dose HSP90 inhibitors maybe similarly efficacious when combined
with CTLA-4 immuno-therapies such as Tremlimumab and
Ipilimumab.
[0204] Cancers that have been clinically validated to show
sensitivity to PD-1/PD-L1 inhibitors such as melanoma, Hodgkin
lymphoma, NSCLC, bladder cancer, renal cell carcinoma may show
equal sensitivity to lower dose HSP90 inhibitors.
Example 2: HSP90 Inhibitors from Different Chemical Classes Each
Synergize with the Selective BCL-2 Inhibitor, Venetoclax, in their
Anti-Cancer Activity
[0205] The ability of HSP90 inhibitors to act synergistically with
the selective BCL-2 inhibitor, venetoclax, was examined using HSP90
inhibitors representing six different chemical scaffolds. These
included the purine-like inhibitor, MPC-0767, a resorcinol
derivative, AT-13387, a geldanamycin derivative, tanespimycin, a
pyrazolopyridine derivative, TAS-116, a dihydroindazolone
derivative, SNX-5422, and a tropane derivative, XL888. Selecting
inhibitors with such diverse chemical scaffolds increases the
likelihood that any observations made are due to on-target
activity, i.e. inhibition of HSP90, because any off-target effects
would be expected to vary between different inhibitor molecules,
and especially between different inhibitor molecules based on
different chemical scaffolds.
[0206] We also sought to determine whether the HSP90 inhibitors act
synergistically with the BCL-2 inhibitor, venetoclax, in different
cancer types. To do this we used four different cancer cell lines
representing each of the three classes of haematopoietic and
lymphoid cancers, leukemias, lymphomas, and myelomas. For
leukemias, we used an acute myeloid leukemia (AML) cell line,
MV-4-11; for lymphomas, we used a diffuse large B cell lymphoma
(DLBCL) cell line, OCI-LY-19, and a mantle cell lymphoma cell line,
Z138; and for myelomas we used the multiple myeloma cell line,
KMS-28.
[0207] Cells were treated with venetoclax combined with each of the
six HSP90 inhibitors at the concentrations shown in Table 2.
TABLE-US-00002 TABLE 2 Cell lines, HSP90 inhibitors, and drug
concentrations (nM) tested. MV-4-11 OCI-LY-19 Z138 KMS-28
Venetoclax 0.78-100 0.78-100 0.78-100 3.9-500 MPC-0767 78-10000
78-10000 78-10000 78-10000 Tanespimycin 7.8-1000 78-10000 7.8-1000
78-10000 TAS-116 7.8-1000 7.8-1000 7.8-1000 7.8-1000 AT-13387
7.8-1000 7.8-1000 7.8-1000 7.8-1000 SNX-5422 7.8-1000 7.8-1000
7.8-1000 7.8-1000 XL-888 7.8-1000 7.8-1000 7.8-1000 7.8-1000
[0208] After 72 hour treatment with the indicated drug
combinations, cell viability was assessed using CellTiter-Glo.
Isobologram analysis was performed to determine synergistic
interactions. Briefly, normalized isobolograms were used to depict
the drug interactions observed across different cell lines at a
dose effect of 75% (EC75). The absolute EC75 for each single agent
and drug combination was calculated using the R package DRC (Ritz
et al., Dose-Response Analysis Using R. PLoS One. 2015.
10(12):e0146021; R Core Team, A language and environment for
statistical computing. R Foundation for Statistical Computing,
Vienna, Austria. 2017). Next, we normalized the EC75 of the drug
combination with respect to corresponding single agent EC75 values.
In cases when single agent treatments did not reach EC75, then the
relative EC75 was used based on the projected value of the fitted
drug response curve. When the relative EC75 was higher than the
maximum concentration tested, we used the maximum concentration
tested as the default value, to allow analysis across all drugs and
conditions.
[0209] As shown in FIG. 11A-F, each of the HSP90 inhibitors
demonstrated synergistic activity with venetoclax in all of the
cell lines. This is evidenced in the figure by the location of each
data point being below the line of additivity, which is shown in
each plot as a dashed line.
[0210] These findings indicate that HSP90 inhibitors generally are
able to act synergistically with venetoclax to inhibit cell
viability in haematopoietic and lymphoid cancers. These data also
indicate that the synergistic anti-cancer activity is due to
on-target inhibition of HSP90, as opposed to off-target
effects.
Example 3: BCL-2 Abundance is Predictive of Synergy Between
MPC-0767 and Venetoclax in Multiple Cancers
[0211] Venetoclax sensitivity has been shown to correlate with the
abundance of its molecular target, BCL-2 protein (Pan et al.,
Selective BCL-2 inhibition by ABT-199 causes on-target cell death
in acute myeloid leukemia. Cancer Discov. 2014; 4(3):362-75).
[0212] To test whether BCL-2 abundance is predictive of the synergy
between HSP90 inhibitors and venetoclax observed here, we analyzed
BCL-2 abundance in cell lines representing acute myeloid leukemia
(MV-4-11, MOLM-16, TUR and U937), multiple myeloma (KMS-28),
diffuse large B cell lymphoma (OCI-LY-19) and mantle cell lymphoma
(Z138) under basal conditions using flow cytometry.
[0213] In parallel, the same cell lines representing acute myeloid
leukemia, multiple myeloma, diffuse large B cell lymphoma and
mantle cell lymphoma were treated with venetoclax for 72 hours and
cell viability determined by CellTiter-Glo. EC50 values were
determined as described above.
[0214] These data showed a trend between basal levels of BCL-2 and
venetoclax sensitivity (FIG. 12). We next determined whether the
observed HSP90 inhibitor and venetoclax synergy depended on basal
BCL-2 levels. Each cell line was treated with a combination of
MPC-0767 and venetoclax and viability assayed 72 hours later using
CellTiter-Glo. Synergy was determined as described above. As shown
in FIG. 12, synergy with MPC-0767 and venetoclax was observed in 4
out of 4 cell lines with high BCL-2 levels. In contrast, no synergy
was observed in the three cell lines with low BCL-2 levels. These
findings suggest that cancers with a reliance on BCL-2 may be
especially susceptible to treatment with a combination of an HSP90
inhibitor and a BCL-2 pathway inhibitor.
Example 4: MPC-0767 Blocks Interferon-Induced PD-L1 Expression
[0215] The release of interferon-gamma (IFN-.gamma.) from T-cells
plays a key role in the host immune response to infections. Yet
this released IFN-.gamma. also provides a mechanism by which tumor
cells can evade the immune system through induction of programmed
death-ligand-1 (PD-L1).
[0216] To ascertain whether MPC-0767 can block IFN-.gamma.-induced
PD-L1 expression in AML cells, four AML cell lines harboring WT
FLT3 (n=2) or FLT3-ITD (n=2) were treated with human IFN-.gamma.
(50 ng/ml) alone, MPC-0767 (2 .mu.M) alone or the combination for
24 hours. Cells were harvested to determine PD-L1 cell surface
expression by flow cytometry. Cells were also stained with a
viability stain to gate on viable cells and exclude any dead cells.
As shown in FIG. 13A-D, all cell lines induced PD-L1 cell surface
expression in response to IFN-.gamma. treatment (6-25 fold). While
MPC-0767 alone did not significantly reduce basal PD-L1 cell
surface expression, in combination with IFN-.gamma., MPC-0767
reduced the IFN-.gamma.-induced PD-L1 cell surface expression in
all cell lines (range: 43-83% reduction).
[0217] These data demonstrate that MPC-0767 displays
immuno-modulatory activity in AML cells by blocking
IFN-.gamma.-induced PD-L1 expression.
* * * * *