U.S. patent application number 14/687768 was filed with the patent office on 2015-11-12 for combination therapies.
The applicant listed for this patent is INFINITY PHARMACEUTICALS, INC.. Invention is credited to Jeffery L. Kutok, Howard M. Stern.
Application Number | 20150320755 14/687768 |
Document ID | / |
Family ID | 53008913 |
Filed Date | 2015-11-12 |
United States Patent
Application |
20150320755 |
Kind Code |
A1 |
Kutok; Jeffery L. ; et
al. |
November 12, 2015 |
COMBINATION THERAPIES
Abstract
Provided herein are pharmaceutical compositions comprising a
phosphatidylinositol 3-kinase inhibitor or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor or a
pharmaceutically acceptable form thereof. Also provided herein are
methods for treating cancer comprising administration the
compositions, and uses of the compositions, e.g., for the treatment
of cancer.
Inventors: |
Kutok; Jeffery L.; (Natick,
MA) ; Stern; Howard M.; (Waban, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
INFINITY PHARMACEUTICALS, INC. |
Cambridge |
MA |
US |
|
|
Family ID: |
53008913 |
Appl. No.: |
14/687768 |
Filed: |
April 15, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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62042691 |
Aug 27, 2014 |
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62042681 |
Aug 27, 2014 |
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61980549 |
Apr 16, 2014 |
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Current U.S.
Class: |
514/235.8 ;
514/253.04; 514/263.21; 514/263.22 |
Current CPC
Class: |
A61K 31/437 20130101;
A61K 31/496 20130101; A61P 35/00 20180101; A61K 31/5377 20130101;
A61K 45/06 20130101; A61K 31/52 20130101; A61K 31/496 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 31/437 20130101;
A61K 31/52 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/52 20060101
A61K031/52; A61K 31/5377 20060101 A61K031/5377; A61K 45/06 20060101
A61K045/06; A61K 31/496 20060101 A61K031/496 |
Claims
1. (canceled)
2. A method of treating a cancer in a subject comprising
administering to the subject a synergistic combination of a PI3K
inhibitor, or a pharmaceutically acceptable form thereof, wherein
the PI3K inhibitor is
(S)-3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)--
one or
(S)-2-(1-(9H-purin-6-ylamino)propyl)-5-fluoro-3-phenylquinazolin-4(-
3H)-one; and a Bcl-2 inhibitor, or a pharmaceutically acceptable
form thereof.
3. A composition comprising a synergistic combination of a PI3K
inhibitor, or a pharmaceutically acceptable form thereof, wherein
the PI3K inhibitor is
(S)-3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)--
one or
(S)-2-(1-(9H-purin-6-ylamino)propyl)-5-fluoro-3-phenylquinazolin-4(-
3H)-one; and a Bcl-2 inhibitor, or a pharmaceutically acceptable
form thereof.
4. The method of claim 2, wherein the combination is synergistic as
indicated by a combination index value that is less than 1, less
than 0.7, or less than 0.5 for the combination of the PI3K
inhibitor and the Bcl-2 inhibitor.
5-6. (canceled)
7. The method of claim 2, wherein the combination index value is
assessed at 50% or more inhibition.
8. The method of claim 2, wherein the combination index value is
assessed at 50% or more growth inhibition.
9-10. (canceled)
11. The method of claim 2, wherein PI3K inhibitor is at an amount
sufficient to reach maximum plasma concentration at steady state
(Cmaxss) at about 1000 ng/mL to about 5000 ng/mL; and the Bcl-2
inhibitor is administered at an amount to reach Cmaxss at about 0.1
.mu.g/mL to about 1000 .mu.g/mL.
12. The method of claim 2, wherein PI3K inhibitor is at an amount
sufficient to reach an area under the plasma concentration-time
curve at steady-state (AUCss) at about 5000 ng/mL*hr to about 10000
ng/mL*hr; and the Bcl-2 inhibitor is administered at an amount to
reach an AUCss at about 0.1 ng/mL*hr to about 10000 ng/mL*hr.
13. The method of claim 2, wherein the PI3K inhibitor is
##STR00010## or a pharmaceutically acceptable form thereof.
14. The method of claim 2, wherein the PI3K inhibitor is
##STR00011## or a pharmaceutically acceptable form thereof.
15. The method of claim 2, wherein the Bcl-2 inhibitor is ABT-199,
ABT-263, ABT-737, G3139 (genasense or oblimersen), GX15-070
(obatoclax mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101),
antimycin A, apogossypol, S44563, or a combination thereof.
16. The method of claim 15, wherein the Bcl-2 inhibitor is
ABT-199.
17. The method of claim 16, wherein the composition comprises 50 to
350 mg of ABT-199.
18. The method of claim 16, wherein the composition comprises about
400 mg, about 325 mg, about 150 mg, or about 75 mg of ABT-199.
19-21. (canceled)
22. The method of claim 2, wherein the PI3K inhibitor is Compound 1
or a pharmaceutically acceptable form thereof and the molar ratio
of Compound 1, or the pharmaceutically acceptable form thereof, to
the Bcl-2 inhibitor, or pharmaceutically acceptable form thereof,
is in the range of from about 50:1 to about 1:50, from about 10:1
to about 1:10, or from about 1:3 to about 1:7.
23. (canceled)
24. The method of claim 2, wherein the PI3K inhibitor is Compound
1, or a pharmaceutically acceptable form thereof, at an amount in
the range of from about 0.01 mg to about 75 mg and the Bcl-2
inhibitor, or a pharmaceutically acceptable form thereof, at an
amount of in the range of from about 0.01 mg to about 1100 mg.
25. (canceled)
26. The method of claim 2, wherein the PI3K inhibitor, or a
pharmaceutically acceptable form thereof, and the Bcl-2 inhibitor,
or a pharmaceutically acceptable form thereof, are in a single
dosage form or in separate dosage forms.
27-28. (canceled)
29. The method of claim 2, wherein the concentration of the PI3K
inhibitor that is required to achieve inhibition is at least 20%
lower when the PI3K inhibitor is administered in combination with
the Bcl-2 inhibitor than when the PI3K inhibitor is administered
alone.
30. The method of claim 2, wherein the concentration of the Bcl-2
inhibitor that is required to achieve inhibition is at least 20%
lower when the Bcl-2 inhibitor is administered in combination with
PI3K inhibitor than when the Bcl-2 inhibitor is administered
alone.
31.-36. (canceled)
37. The method of claim 2, wherein the anti-cancer effect provided
by the composition is greater than the anti-cancer effect provided
by a monotherapy with the same dose of the PI3K inhibitor or
pharmaceutically acceptable form thereof as is included in the
composition.
38. The method of claim 2, wherein the anti-cancer effect provided
by the composition is at least 2 fold greater, at least 3 fold
greater, at least 5 fold greater, or at least 10 fold greater than
the anti-cancer effect provided by the monotherapy with the PI3K
inhibitor or pharmaceutically acceptable form thereof.
39-41. (canceled)
42. The method of claim 3, wherein the PI3K inhibitor, or a
pharmaceutically acceptable form, is administered concurrently
with, subsequent to, or prior to the Bcl-2 inhibitor.
43-44. (canceled)
45. The method of claim 2, which delays resistance of the cancer to
the PI3K inhibitor or reduces the risk that the cancer becomes
resistant to the PI3K inhibitor.
46. (canceled)
47. The method of claim 45, wherein the cancer does not become
resistant to the PI3K inhibitor for at least 12, 18, or 24
months.
48. The method of claim 2, which prolongs remission in the
subject.
49. The method of claim 48, wherein the subject experiences
remission for at least 12 months.
50. (canceled)
51. The method of claim 2, wherein the subject experiences complete
remission.
52. The method of claim 2, which results in a reduction in the
level of minimal residual disease (MRD).
53. The method of claim 2, wherein the subject has substantially no
detectable MRD.
54-58. (canceled)
59. A method of delaying or decreasing resistance of a subject
having a cancer, comprising administering to the subject a
synergistic amount of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, thereby delaying or
decreasing resistance.
60-61. (canceled)
62. A method of reducing the level of minimal residual disease
(MRD) compared to a reference value in a subject having a cancer,
comprising administering to the subject a synergistic amount of a
PI3K inhibitor, or a pharmaceutically acceptable form thereof, and
a Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
thereby reducing the level of MRD in the subject.
63. A method of treating a cancer or tumor in a subject,
comprising: acquiring a value for one or more of: the presence,
absence, amount or level of an alteration or biomarker chosen from
one, two, three, four, five, six, seven, eight, nine, 10, 11, 12,
13, 14, 15, or all of: an STK11 copy number, TSC1 copy number, TSC2
copy number, TP53 copy number, PTEN copy number, CBFA2T3 copy
number, YWHAE copy number, PER1 copy number, GAS7 copy number,
FSTL3 copy number, USP6 copy number, MAP2K4 copy number, EGFR copy
number, a BCR pathway mutation a p53 pathway mutation, or a MAPK
pathway mutation, or any combination thereof, and responsive to
said value, administering to the subject a composition according to
claim 3.
64-67. (canceled)
68. The method of claim 63, wherein one, two, three, four, five,
six, seven, eight, nine, 10, 11, 12, 13, or all of the following is
indicative of decreased responsiveness of the cancer or tumor, or
the subject, to the treatment: (i) a copy number loss of STK11;
(ii) a copy number loss of TSC1 or TSC2, or both; (iii) a copy
number loss of TP53; (iv) a copy number loss of PTEN; (v) a copy
number loss of CBFAT2T3; (vi) a copy number loss of YWHAE; (vii) a
copy number loss of PER1; (viii) a copy number loss of GAS7; (ix) a
copy number loss of FSTL3; (x) a copy number loss of USP6; (xi) a
copy number loss of MAP2K4; (xii) a BCR pathway mutation (xiii) a
p53 pathway mutation; or (xiv) a MAPK pathway mutation.
69. The method of claim 2, wherein the cancer is of hematopoietic
origin.
70. The method of claim 69, wherein the cancer is a lymphoma or
leukemia.
71. The method of claim 70, wherein the cancer is acute
lymphoblastic leukemia (ALL), T-cell ALL (T-ALL), B-cell ALL
(B-ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia
(CLL), chronic myelogenous leukemia (CML), blast phase CML, small
lymphocytic lymphoma (SLL), CLL/SLL, blast phase CLL, Hodgkin
lymphoma (HL), non-Hodgkin lymphoma (NHL), B-cell NHL, T-cell NHL,
indolent NHL (iNHL), diffuse large B-cell lymphoma (DLBCL), mantle
cell lymphoma (MCL), aggressive B-cell NHL, B-cell lymphoma (BCL),
Richter's syndrome (RS), T-cell lymphoma (TCL), peripheral T-cell
lymphoma (PTCL), cutaneous T-cell lymphoma (CTCL), transformed
mycosis fungoides, Sezary syndrome, anaplastic large-cell lymphoma
(ALCL), follicular lymphoma (FL), Waldenstrom macroglobulinemia
(WM), lymphoplasmacytic lymphoma, Burkitt lymphoma, multiple
myeloma (MM), myeloproliferative disorder (MPD), myelofibrosis
(MF), chronic myelomonocytic leukemia (CMML), angioimmunoblastic
lymphoma, cutaneous lymphoma, myeloma, plasmacytoma, diffuse large
B-cell lymphoma activated B-cell like, or diffuse large B-cell
lymphoma germinal center B-cell-like.
72-87. (canceled)
Description
[0001] This application claims priority to U.S. Ser. No.
61/980,549, filed Apr. 16, 2014, U.S. Ser. No. 62/042,691 filed
Aug. 27, 2014, and U.S. Ser. No. 62/042,681 filed Aug. 27, 2014,
the contents of which are incorporated herein by reference in their
entireties.
BACKGROUND
[0002] The phosphoinositide 3-kinases (PI3Ks) signaling pathway is
one of the most highly mutated systems in human cancers. PI3Ks are
members of a unique and conserved family of intracellular lipid
kinases that phosphorylate the 3'-OH group on phosphatidylinositols
or phosphoinositides. The PI3K family comprises 15 kinases with
distinct substrate specificities, expression patterns, and modes of
regulation. The class I PI3Ks (p110.alpha., p110.beta.,
p110.delta., and p110.gamma.) are typically activated by tyrosine
kinases or G-protein coupled receptors to generate
phosphatidylinositol (3,4,5)-trisphosphate (PIP3), which engages
downstream effectors such as those in the AKT/PDK1 pathway, mTOR,
the Tec family kinases, and the Rho family GTPases. The class II
and III PI3Ks play a key role in intracellular trafficking through
the synthesis of phosphatidylinositol 3-bisphosphate (PI(3)P) and
phosphatidylinositol (3,4)-bisphosphate (PI(3,4)P2). The PI3Ks are
protein kinases that control cell growth (mTORC1) or monitor
genomic integrity (ATM, ATR, DNA-PK, and hSmg-1).
[0003] There are four mammalian isoforms of class I PI3Ks:
PI3K-.alpha., .beta., .delta. (class Ia PI3Ks) and PI3K-.gamma. (a
class Ib PI3K). These enzymes catalyze the production of PIP3,
leading to activation of downstream effector pathways important for
cellular survival, differentiation, and function. PI3K-.alpha. and
PI3K-.beta. are widely expressed and are important mediators of
signaling from cell surface receptors. PI3K-.alpha. is the isoform
most often found mutated in cancers and has a role in insulin
signaling and glucose homeostasis (Knight et al. Cell (2006)
125(4):733-47; Vanhaesebroeck et al. Current Topic Microbiol.
Immunol. (2010) 347:1-19). PI3K-.beta. is activated in cancers
where phosphatase and tensin homolog (PTEN) is deleted. Both
isoforms are targets of small molecule therapeutics in development
for cancer.
[0004] PI3K-.delta. and -.gamma. are preferentially expressed in
leukocytes and are important in leukocyte function. These isoforms
also contribute to the development and maintenance of hematologic
malignancies (Vanhaesebroeck et al. Current Topic Microbiol.
Immunol. (2010) 347:1-19; Clayton et al. J Exp Med. (2002)
196(6):753-63; Fung-Leung Cell Signal. (2011) 23(4):603-8;
Okkenhaug et al. Science (2002) 297(5583):1031-34). PI3K-.delta. is
activated by cellular receptors (e.g., receptor tyrosine kinases)
through interaction with the Sarc homology 2 (SH2) domains of the
PI3K regulatory subunit (p85), or through direct interaction with
RAS.
SUMMARY
[0005] The present invention provides, at least in part,
compositions and methods comprising a PI3K inhibitor in combination
with a Bcl-2 inhibitor. In one embodiment, it has been discovered a
combination of a PI3K inhibitor (e.g., Compound 1) and a Bcl-2
inhibitor (e.g., ABT-199 or ABT-263) has a synergistic effect in
treating a cancer (e.g., in reducing cancer cell growth or
viability, or both). In certain embodiments, the combination of a
PI3K inhibitor and a Bcl-2 inhibitor can allow the PI3K inhibitor,
the Bcl-2 inhibitor, or both, to be administered at a lower dosage,
e.g., a lower dosage than would be required to achieve the same
therapeutic effect if the PI3K inhibitor or Bcl-2 inhibitor were
administered as a monotherapy. In some embodiments, the combination
can allow the PI3K inhibitor, the Bcl-2 inhibitor, or both, to be
administered at a lower frequency than if the PI3K inhibitor or
Bcl-2 inhibitor were administered as a monotherapy. Such
combinations provide advantageous effects, e.g., in reducing (e.g.,
inhibiting, preventing, delaying, and/or decreasing the likelihood
of occurrence of) one or more of: a side effect, toxicity, or
resistance (e.g., acquired resistance) that would otherwise be
associated with administration of a higher dose of the agent(s),
e.g., when the agent is administered as a monotherapy.
[0006] In one aspect, provided herein are compositions, e.g.,
pharmaceutical compositions comprising a PI3K inhibitor (e.g., one
or more PI3K inhibitors), or a pharmaceutically acceptable form
thereof, in combination with a Bcl-2 inhibitor (e.g., one or more
Bcl-2 inhibitors), or a pharmaceutically acceptable form thereof.
By "in combination with" it is not intended to imply that the Bcl-2
inhibitor and the PI3K inhibitor must be administered at the same
time and/or formulated for delivery together, although these
methods of delivery are within the scope of this disclosure.
[0007] In some embodiments, the PI3K inhibitor and the Bcl-2
inhibitor are present in a single composition.
[0008] In some embodiments, the PI3K inhibitor and the Bcl-2
inhibitor are present in two or more different compositions (e.g.,
as separate dosage forms). In some embodiments, the PI3K inhibitor
and the Bcl-2 inhibitor are formulated for administration via the
same administration route. In some embodiments, the PI3K inhibitor
and the Bcl-2 inhibitor are formulated for administration via
different administration routes.
[0009] In some embodiments, the composition (e.g., one or more
compositions or dosage forms) comprising the combination of PI3K
inhibitor and the Bcl-2 inhibitor) is synergistic, e.g., the
combination has a synergistic effect, e.g., a synergistic effect in
treating a cancer (e.g., in reducing cancer cell growth or
viability, or both). In certain embodiments, the amount or dosage
of the PI3K inhibitor, the Bcl-2 inhibitor, or both, present in the
composition(s) does not exceed the level at which each agent is
used individually, e.g., as a monotherapy. In certain embodiments,
the amount or dosage of the PI3K inhibitor, the Bcl-2 inhibitor, or
both, present in the composition(s) is lower (e.g., at least 20%,
at least 30%, at least 40%, or at least 50%) than the amount or
dosage of each agent used individually, e.g., as a monotherapy. In
other embodiments, the amount or dosage of the PI3K inhibitor, the
Bcl-2 inhibitor, or both, present in the composition(s) that
results in a desired effect (e.g., treatment of cancer, achieve
inhibition e.g., 50% inhibition, achieve growth inhibition e.g.,
50% growth inhibition, achieve a therapeutic effect) is lower
(e.g., at least 20%, at least 30%, at least 40%, or at least 50%
lower) than the amount or dosage of each agent used individually,
e.g., as a monotherapy. In certain embodiments, the frequency of
administration of the PI3K inhibitor that achieves a therapeutic
effect is lower (e.g., at least 20%, 30%, 40%, or 50% lower), when
the PI3K inhibitor is administered in combination with the Bcl-2
inhibitor than when the PI3K inhibitor is administered alone. In
some embodiments, the frequency of administration of the Bcl-2
inhibitor that achieves a therapeutic effect is lower (e.g., at
least 20%, 30%, 40%, or 50% lower), when the Bcl-2 inhibitor is
administered in combination with PI3K inhibitor than when the Bcl-2
inhibitor is administered alone.
[0010] In another aspect, featured herein is a method of treating
(e.g., inhibiting, reducing, ameliorating, managing, or preventing)
a cancer in a subject. The method includes administering to the
subject a PI3K inhibitor (e.g., one or more PI3K inhibitors), or a
pharmaceutically acceptable form thereof, in combination with a
Bcl-2 inhibitor (e.g., one or more Bcl-2 inhibitors), or
pharmaceutically acceptable form thereof.
[0011] The combination of the PI3K inhibitor and the Bcl-2
inhibitor can be administered together in a single composition or
administered separately in two or more different compositions,
e.g., pharmaceutical compositions or dosage forms as described
herein. The administration of the PI3K inhibitor and the Bcl-2
inhibitor can be in any order. The PI3K inhibitor can be
administered concurrently with, prior to, or subsequent to, the
Bcl-2 inhibitor. In one embodiment, the Bcl-2 inhibitor is
administered to a subject at least 5 minutes, 15 minutes, 30
minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours,
or 24 hours before the PI3K inhibitor (e.g., Compound 1), or a
pharmaceutically acceptable form thereof, is administered. In
another embodiment, the Bcl-2 inhibitor is administered
concurrently with the PI3K inhibitor (e.g., Compound 1), or a
pharmaceutically acceptable form thereof, e.g., in a single dosage
form or separate dosage forms. In yet another embodiment, the Bcl-2
inhibitor is administered to the subject at least 5 minutes, 15
minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours,
12 hours, or 24 hours after the PI3K inhibitor (e.g., Compound 1),
or a pharmaceutically acceptable form thereof, is administered. In
some embodiments, the PI3K inhibitor and the Bcl-2 inhibitor are
administered with a timing that results in both inhibitors being
present at therapeutic levels at the same time in the patient.
[0012] In some embodiments, the combination of the PI3K inhibitor
and the Bcl-2 inhibitor is additive, e.g., the effect of the
combination is similar to their individual effects added
together.
[0013] In some embodiments, the combination of PI3K inhibitor and
the Bcl-2 inhibitor) is synergistic, e.g., has a synergistic
effect, e.g., a synergistic effect in treating a cancer (e.g., in
reducing cancer cell growth or viability, or both).
[0014] In some embodiments, the amount or dosage of the PI3K
inhibitor, the Bcl-2 inhibitor, or both, used in combination does
not exceed the level at which each agent is used individually,
e.g., as a monotherapy. In certain embodiments, the amount or
dosage of the PI3K inhibitor, the Bcl-2 inhibitor, or both, used in
combination is lower (e.g., at least 20%, at least 30%, at least
40%, or at least 50% lower) than the amount or dosage of each agent
used individually, e.g., as a monotherapy. In other embodiments,
the amount or dosage of the PI3K inhibitor, the Bcl-2 inhibitor, or
both, used in combination that results in treatment of cancer is
lower (e.g., at least 20%, at least 30%, at least 40%, or at least
50% lower) than the amount or dosage of each agent used
individually, e.g., as a monotherapy. In certain embodiments, the
frequency of administration of the PI3K inhibitor, the Bcl-2
inhibitor, or both, used in combination that results in treatment
of cancer is lower (e.g., at least 20%, 30%, 40%, or 50% lower),
than the frequency of administration of each agent used
individually, e.g., as a monotherapy.
[0015] The combination of PI3K inhibitor and the Bcl-2 inhibitor
can be administered during periods of active disorder, or during a
period of remission or less active disease. The combination can be
administered before a third treatment or procedure (e.g., radiation
or surgery), concurrently with the third treatment, post-treatment,
or during remission of the disorder.
[0016] In another aspect, featured herein is a method of inhibiting
the growth or the viability, or both, of a cancer cell. The method
includes contacting the cancer cell with a PI3K inhibitor (e.g.,
one or more PI3K inhibitors), or a pharmaceutically acceptable form
thereof, in combination with a Bcl-2 inhibitor (e.g., one or more
Bcl-2 inhibitors), or pharmaceutically acceptable form thereof. In
certain embodiments, the method is for use in vitro. In certain
embodiments, the method is for use or in vivo, e.g., in an animal
subject or as part of a therapeutic protocol.
[0017] The contacting of the cell with the PI3K inhibitor and the
Bcl-2 inhibitor can be in any order. In certain embodiments, the
cell is contacted with the PI3K inhibitor concurrently, prior to,
or subsequent to, the Bcl-2 inhibitor. In certain embodiments, the
combination of the PI3K inhibitor and the Bcl-2 inhibitor is
synergistic, e.g., has a synergistic effect in reducing cancer cell
growth or viability, or both. In some embodiments, the amount or
dosage of the PI3K inhibitor, the Bcl-2 inhibitor, or both, used in
combination does not exceed the level at which each agent is used
individually, e.g., as a monotherapy. In certain embodiments, the
amount or dosage of the PI3K inhibitor, the Bcl-2 inhibitor, or
both, used in combination is lower (e.g., at least 20%, at least
30%, at least 40%, or at least 50% lower) than the amount or dosage
of each agent used individually, e.g., as a monotherapy. In other
embodiments, the amount or dosage of the PI3K inhibitor, the Bcl-2
inhibitor, or both, used in combination that results in a reducing
cancer cell growth or viability, or both is lower (e.g., at least
20%, at least 30%, at least 40%, or at least 50% lower) than the
amount or dosage of each agent used individually, e.g., as a
monotherapy.
[0018] In another aspect, the present disclosure provides a
synergistic combination of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, for use in treating
cancer. In another aspect, the present disclosure provides a
synergistic combination of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, for use in a medicament.
In another aspect, the present disclosure provides a use of a
synergistic combination of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, for treating cancer. In
another aspect, the present disclosure provides a use of a
synergistic combination of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof for the manufacture of a
medicament for treating cancer.
[0019] Additional embodiments of the compositions and/or methods
described herein include one or more of the following:
[0020] The compositions described herein can be used, e.g., for
treatment of a cancer described herein and/or for inhibiting the
growth or viability of a cancer cell in vitro or in vivo.
[0021] In certain embodiments, the combination of the PI3K
inhibitor and the Bcl-2 inhibitor used in the compositions and
methods described herein is synergistic, e.g., as indicated by a
combination index value that is less than 1 for the combination of
the PI3K inhibitor and the Bcl-2 inhibitor. In certain embodiments,
the combination is synergistic as indicated by a combination index
value that is less than 0.7 for the combination of the PI3K
inhibitor and the Bcl-2 inhibitor. In some embodiments, the
combination is synergistic as indicated by a combination index
value that is less than 0.5 for the combination of the PI3K
inhibitor and the Bcl-2 inhibitor. In some embodiments, the
combination is synergistic as indicated by a combination index
value that is less than 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, or 0.1 for
the combination of the PI3K inhibitor and the Bcl-2 inhibitor. In
some embodiments, the combination of the PI3K inhibitor and the
Bcl-2 inhibitor used in the compositions and methods described
herein is additive, e.g., as indicated by a combination index value
that is equal to about 1 for the combination of the PI3K inhibitor
and the Bcl-2 inhibitor. In some embodiments, the combination index
value is assessed at 50% or more inhibition or growth inhibition.
In certain embodiments, the combination index value is assessed at
50% inhibition, e.g., as described herein in the Examples. In some
embodiments, the combination index value is assessed at 50%
inhibition or growth inhibition, e.g., as described herein in the
Examples. In some embodiments, the combination index value is
assessed at 85% inhibition or growth inhibition, e.g., as described
herein in the Examples. In some embodiments, the combination index
value is assessed at 110% growth inhibition, e.g., as described
herein in the Examples. In some embodiments, the combination index
value is assessed at 140% growth inhibition, e.g., as described
herein in the Examples. In some embodiments, the combination index
value is assessed at 10%, 20%, 30%, 40%, 50%, 60%, 60%, 70%, 80%,
90%, 100% inhibition. In some embodiments, the combination index
value is assessed at 10%, 20%, 30%, 40%, 50%, 60%, 60%, 70%, 80%,
90%, 100%, 110%, 120%, 130%, 140%, or 150% or more growth
inhibition. In some embodiments, the combination index value is
calculated as described herein.
[0022] In other embodiments, the combination of the PI3K inhibitor
and the Bcl-2 inhibitor used in the compositions and methods
described herein is synergistic, e.g., as indicated by a synergy
score value of greater than 1, 2, or 3. In certain embodiments, the
combination is synergistic as indicated by a synergy score value
that is 1 or more. In certain embodiments, the combination is
synergistic as indicated by a synergy score value that is 3 or
more. In certain embodiments, the combination is synergistic as
indicated by a synergy score value that is 10 or more. In certain
embodiments, the combination of the PI3K inhibitor and the second
agent used in the compositions and methods described herein is
additive, e.g., as indicated by a synergy score value of zero.
[0023] In one embodiment, the combination is synergistic as
indicated by a synergy score value that is two standard deviations
or three standard deviations of the value of the self-crosses,
which indicates synergy at a confidence level of 95% or 99%,
respectively. In certain embodiments, the synergy score is
calculated as described herein.
[0024] In some embodiments, the anti-cancer effect provided by the
combination of the PI3K inhibitor and the Bcl-2 inhibitor used in
the compositions and methods described herein is greater than the
anti-cancer effect provided by an agent (e.g., the PI3K inhibitor
or the Bcl-2 inhibitor) used individually, e.g., as a monotherapy.
In one embodiment, the anti-cancer effect provided by the
combination of the PI3K inhibitor and the Bcl-2 inhibitor is
greater than the anti-cancer effect provided monotherapy with the
same dose of the PI3K inhibitor. In certain embodiments, the
anti-cancer effect provided by the combination of the PI3K
inhibitor and the Bcl-2 inhibitor is at least 2 fold greater, at
least 3 fold greater, at least 5 fold greater, or at least 10 fold
greater than the anti-cancer effect provided by an agent used
individually, e.g., as a monotherapy (e.g., by a monotherapy with
the same dose of the PI3K inhibitor, or by a monotherapy with the
same dose of the Bcl-2 inhibitor).
[0025] As used herein, a "monotherapy" refers to the use of an
agent individually (also referred to herein as alone), e.g.,
without a second active ingredient to treat the same indication,
e.g., cancer. For example, in this context, the term monotherapy
includes the use of either the PI3K inhibitor or the Bcl-2
inhibitor individually or alone to treat the cancer.
[0026] In some embodiments, the anti-cancer effect provided by the
combination of the PI3K inhibitor and the Bcl-2 inhibitor used in
the compositions and methods described herein is greater than the
anti-cancer effect provided by a monotherapy with the same dose of
the PI3K inhibitor. In certain embodiments, the anti-cancer effect
provided by the combination is at least 2 fold greater, at least 3
fold greater, at least 5 fold greater, or at least 10 fold greater
than the anti-cancer effect provided by the monotherapy with the
same dose of the PI3K inhibitor.
[0027] In some embodiments, the anti-cancer effect of the
combination of the PI3K inhibitor and the Bcl-2 inhibitor used in
the compositions and methods described herein is greater than the
anti-cancer effect provided by a monotherapy with the same dose of
the Bcl-2 inhibitor. In certain embodiments, the anti-cancer effect
of the combination of the PI3K inhibitor and the Bcl-2 inhibitor is
at least 2 fold greater, at least 3 fold greater, at least 5 fold
greater, or at least 10 fold greater than the anti-cancer effect
provided by the monotherapy with the same dose of the Bcl-2
inhibitor.
[0028] In some embodiments, one or more side effects of the PI3K
inhibitor, the Bcl-2 inhibitor, or both, is reduced compared with
the side effects of an agent (e.g., the side effects of either the
PI3K inhibitor or the Bcl-2 inhibitor) when used individually,
e.g., as a monotherapy (e.g., a monotherapy with the PI3K
inhibitor, or by a monotherapy with the Bcl-2 inhibitor), e.g.,
when the monotherapy is administered at a dose that achieves the
same therapeutic effect as the combination.
[0029] In some embodiments, one or more side effects of the PI3K
inhibitor, the Bcl-2 inhibitor, or both is reduced compared with
the side effects of each agent when used individually, e.g., as a
monotherapy (e.g., a monotherapy with the PI3K inhibitor, or by a
monotherapy with the Bcl-2 inhibitor), e.g., when the monotherapy
is administered at a dose that achieves the same therapeutic effect
as the combination.
[0030] In some embodiments, one or more side effects of the
compositions or methods described herein is reduced compared with
the side effects of a monotherapy comprising either the Bcl-2
inhibitor (or pharmaceutically acceptable form thereof) or the PI3K
inhibitor (or pharmaceutically acceptable form thereof), e.g., a
monotherapy at a dose that achieves the same therapeutic effect as
does the combination.
[0031] In some embodiments, said one or more side effects includes
a liver enzyme level, e.g., a liver enzyme level indicative of
toxicity.
[0032] In some embodiments, said one or more side effects include
thrombocytopenia.
[0033] In some embodiments, said one or more side effects include
tumor lysis syndrome.
[0034] In some embodiments, the combination of the PI3K inhibitor
and the Bcl-2 inhibitor used in the compositions and methods
described herein results in a reduction in resistance (e.g., a
decrease in a measure of resistance or a decreased likelihood of
developing resistance), or a delay in the development of
resistance, to at least one of the agents, e.g., resistance (e.g.,
acquired resistance) to the PI3K inhibitor. In certain embodiments,
resistance is assessed at least in part by evaluating minimal
residual disease (MRD).
[0035] In some embodiments, the combination of the PI3K inhibitor
and the Bcl-2 inhibitor used in the compositions and methods
described herein results in a reduction in the level of minimal
residual disease (MRD) that can be detected. In certain
embodiments, the combination of a PI3K inhibitor (e.g. a PI3K
inhibitor described herein) and a Bcl-2 inhibitor (e.g., a Bcl-2
inhibitor described herein) is effective to reduce the level of MRD
in the subject, e.g., below a level previously measured in the
subject (e.g., the level measured before the combination was
administered). In certain embodiments, the combination of a PI3K
inhibitor and a Bcl-2 inhibitor is effective to reduce the level of
MRD in the subject below the level observed during or after
treatment with a monotherapy, e.g., a monotherapy comprising either
the PI3K inhibitor or the Bcl-2 inhibitor. In certain embodiments,
the level of MRD is decreased below the level observed during
treatment with a monotherapy comprising the PI3K inhibitor. In
certain embodiments, the level of MRD is decreased below the level
observed during treatment with a monotherapy comprising the Bcl-2
inhibitor. In certain embodiments, the combination is effective to
reduce the level of MRD below a preselected cutoff value (e.g., 1
malignant cell in 100 normal cells, 1 malignant cell in 1000 normal
cells, 1 malignant cell in 10,000 normal cells, or 1 malignant cell
in 100,000 normal cells). In certain embodiments, the preselected
cutoff value is 1 malignant cell in 1,000 or 10,000 normal cells.
In some embodiments, a subject exhibits MRD negativity (or is
MRD-negative) if the MRD is below a preselected cutoff value (e.g.,
a preselected cutoff value as described herein). In some
embodiments, the level of MRD is not detectable by standard
laboratory methodologies.
[0036] In another aspect, the invention features a method of
treating a cancer in a subject, or a method of decreasing the level
of MRD in a subject having a cancer. The method comprises:
[0037] (a) administering to the subject a PI3K inhibitor (e.g.,
Compound 1), or a pharmaceutically acceptable form thereof, in
combination with a Bcl-2 inhibitor (e.g., at least one Bcl-2
inhibitor) (also referred to as "a first treatment");
[0038] (b) monitoring the level of MRD in the subject, e.g., by one
or more of the methods described herein or known in the art (e.g.,
flow cytometry, sequencing, or PCR); and
[0039] (c) if the subject is MRD negative, e.g., for a time period
after therapy (e.g., at least 1, 2, 3, 6, 9, 12 months), alter the
combination treatment (e.g., reduce the dose or cease the first
treatment).
[0040] In some embodiments, the method further includes monitoring
the subject after altering the combination treatment (e.g., after
reducing the dose or ceasing the first treatment), e.g., for a
period of at least 6 months, 9 months or 12 months. If the MRD
levels increase, e.g., increase above a preselected cutoff value
(e.g., a preselected cutoff value as described herein), administer
a second treatment. In one embodiment the second treatment is a
PI3K inhibitor monotherapy. In another embodiment, the second
treatment is a PI3K inhibitor in combination with a Bcl-2 inhibitor
(e.g., at least one Bcl-2 inhibitor). In another embodiment, the
second treatment is a PI3K inhibitor in combination with a third
agent. In yet another embodiment, the second treatment is a PI3K
inhibitor in combination a Bcl-2 inhibitor and a third agent.
[0041] In another aspect, the invention features a method of
treating a cancer in a subject, or a method of decreasing the level
of MRD detected in a subject having a cancer. The method
comprises:
[0042] (a) administering to the subject a PI3K inhibitor (e.g.,
Compound 1), or a pharmaceutically acceptable form thereof, in
combination with a Bcl-2 inhibitor (e.g., at least one Bcl-2
inhibitor) (also referred to as "a first treatment");
[0043] (b) monitoring the MRD in the subject, e.g., by one or more
methods described herein or known in the art (e.g., flow cytometry,
sequencing, or PCR); and
[0044] (c) stop administering the first treatment (e.g., the
combination) if the level of MRD in the subject is decreases below
a preselected cutoff value (e.g., 1 malignant cell in 100 normal
cells, 1 malignant cell in 1000 normal cells, or 1 malignant cell
in 10,000 normal cells). In some embodiments, the method further
comprises (d) monitoring the level of MRD in the subject, e.g., by
one or more of the methods described herein or known in the art
(e.g., flow cytometry, sequencing, or PCR) and (e) administering a
second treatment (e.g., a monotherapy comprising a PI3K inhibitor,
or administering a further combination comprising the PI3K
inhibitor, or a pharmaceutically acceptable form thereof), if the
level of MRD increases, e.g., increase above a preselected cutoff
value (e.g., 1 malignant cell in 100 normal cells, 1 malignant cell
in 1000 normal cells, or 1 malignant cell in 10,000 normal cells).
Optionally, the method comprises repeating steps (b), (c), (d) and
(e). In one embodiment the second treatment is a PI3K inhibitor
monotherapy. In another embodiment, the second treatment is a PI3K
inhibitor in combination with a Bcl-2 inhibitor (e.g., at least one
Bcl-2 inhibitor). In another embodiment, the second treatment is a
PI3K inhibitor in combination with a third agent. In yet another
embodiment, the second treatment is a PI3K inhibitor in combination
a Bcl-2 inhibitor and a third agent.
[0045] The aforesaid compositions and methods can be used in
combination with a monotherapy (e.g., a monotherapeutic
administration or dose of the PI3K inhibitor, the Bcl-2 inhibitor
or a third agent). In one embodiment, the subject is administered a
monotherapy with a PI3K inhibitor, which can be followed with a
combination composition or method described herein. For example, if
the subject is developing, or is identified as developing, a
decreased responsiveness to a first monotherapy, (e.g., with a PI3K
inhibitor, Bcl-2 inhibitor, or third agent), any of the combination
compositions or methods described herein can be administered. In
certain embodiments, the combination compositions or methods
described herein improve responsiveness (e.g., as indicated by a
decrease in MRD, e.g., a decrease below the level of MRD observed
during treatment with the first monotherapy). Alternatively,
administration of any of the combination compositions or methods
described herein can be followed by administration of a
monotherapy, e.g., with a PI3K inhibitor, Bcl-2 inhibitor, or third
agent.
[0046] In other embodiments, the composition and methods described
herein can include further agents or therapies, including but not
limited to, chemotherapeutics, radiation or surgery.
[0047] In certain embodiments of the compositions and methods
described herein, the PI3K inhibitor is a PI3K delta inhibitor. In
one embodiment, the PI3K inhibitor is a PI3K delta/gamma dual
inhibitor.
[0048] In one embodiment, the PI3K inhibitor is chosen from one or
more of: Compound 1, AMG-319, GSK 2126458, GSK 1059615, GDC-0032,
GDC-0980, GDC-0941, XL147, XL499, XL765, BKM 120, GS1101, CAL 263,
SF1126, PX-866, BEZ235, CAL-120, BYL719, RP6503, RP6530, TGR1202,
INK1117, PX-886, BAY 80-6946, IC87114, Palomid 529, ZSTK474,
PWT33597, TG100-115, GNE-477, CUDC-907, AEZS-136, BGT-226,
PF-05212384, LY3023414, PI-103, LY294002, INCB-040093, CAL-130 or
wortmannin.
[0049] In one embodiment, the PI3K inhibitor is Compound 1, or a
pharmaceutically acceptable form thereof. Compound 1 has the
following structure:
##STR00001##
[0050] In other embodiments, the PI3K inhibitor is GS1101
(CAL-101), or a pharmaceutically acceptable form thereof.
[0051] In one embodiment, the pharmaceutical composition further
comprises one or more pharmaceutically acceptable excipients.
[0052] In some embodiments of the compositions and methods
described herein, the combination of the PI3K inhibitor and the
Bcl-2 inhibitor is therapeutically effective (e.g., synergistically
effective), in treating a cancer in the subject, e.g., for
treatment of a cancer described herein.
[0053] In one embodiment, the cancer to be treating using the
methods or compositions described herein is of hematopoietic
origin. In one embodiment, the cancer is lymphoma or leukemia. In
one embodiment, the cancer is B-cell lymphoma, mantle cell
lymphoma, non-Hodgkin's lymphoma (e.g., non-Hodgkin's B-cell
lymphoma), T-cell lymphoma (e.g., peripheral T-cell lymphoma (PTCL)
and cutaneous T-cell lymphoma (CTCL)), cutaneous lymphoma,
anaplastic large cell lymphoma, multiple myeloma, myeloma, or
plasmacytoma. In one embodiment, the cancer is a multiple
myeloma.
[0054] In one embodiment, the cancer is chronic lymphocytic
leukemia (CLL).
[0055] In other embodiments, the cancer has a high level of Bcl-2
expression. In one embodiment, the cancer is a diffuse large B-cell
lymphoma with a high level of Bcl-2 expression.
[0056] In one embodiment, the cancer is diffuse large B-cell
lymphoma, follicular lymphoma, T-cell lymphoma, B-cell lymphoma,
mantle cell lymphoma, non-Hodgkin B-cell lymphoma, non-Hodgkin
T-cell lymphoma, indolent non-Hodgkin lymphoma, cutaneous lymphoma,
anaplastic large cell lymphoma, multiple myeloma, myeloma, or
plasmacytoma.
[0057] In other embodiments, the cancer is a non-Hodgkin's
lymphoma. In certain embodiments, the cancer is a B cell
non-Hodgkin's lymphoma. In certain embodiments, the non-Hodgkin's
lymphoma is a diffuse large B-cell lymphoma. In certain
embodiments, the non-Hodgkin's lymphoma is a diffuse large B-cell
lymphoma activated B-cell like or a diffuse large B-cell lymphoma
germinal center B-cell-like. In certain embodiments, the cancer is
an indolent non-Hodgkin's lymphoma, e.g., a follicular lymphoma. In
certain embodiments, the cancer is a mantle cell lymphoma. In
certain embodiments, the cancer is a T-cell non-Hodgkin's
lymphoma.
[0058] In one embodiment, the subject is a mammal, e.g., a human.
In one embodiment, the subject is at risk or suffers from a cancer,
e.g., a cancer described herein.
[0059] In one embodiment, the method delays resistance of the
cancer, e.g., to a therapeutic agent, e.g., to the PI3K inhibitor
such as Compound 1, or to the Bcl-2 inhibitor. In one embodiment,
the method reduces the risk that the cancer becomes resistant,
e.g., to a therapeutic agent, e.g., to the PI3K inhibitor such as
Compound 1, or to the Bcl-2 inhibitor. In one embodiment, the
cancer does not become resistant (e.g., to the PI3K inhibitor) for
at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, or 36
months. In one embodiment, the method prolongs remission (e.g.,
complete remission or partial remission) in the subject. In one
embodiment, the subject experiences remission (e.g., complete
remission or partial remission) for at least 1, 2, 3, 4, 5, 6, 7,
8, 9, 10, 11, 12, 18, 24, 30, or 36 months. In one embodiment, the
method increases the likelihood that the subject experiences
complete remission. In one embodiment, the subject experiences
complete remission. In one embodiment, the method results in a
reduction in the level of minimal residual disease (MRD). In one
embodiment, the subject has substantially no detectable MRD. In
certain embodiments, the subject displays one or more of these
characteristics (e.g., remission) after treatment with the PI3K
inhibitor and the Bcl-2 inhibitor for a therapeutically effective
period of time, e.g., at least 1, 2, 3, or 4 weeks, or 1, 2, 4, 6,
9, or 12 months.
[0060] In one embodiment, the subject is resistant or refractive to
a treatment, e.g., a treatment with a PI3K inhibitor (e.g.,
Compound 1 or GS1101). In one embodiment, the subject is identified
as having a decreased susceptibility (e.g., resistance or acquired
resistance) to a monotherapy treatment with a PI3K inhibitor (e.g.,
Compound 1 or GS1101), or a pharmaceutically acceptable form
thereof.
[0061] In some embodiments of the compositions and methods
described herein, the PI3K inhibitor and the Bcl-2 inhibitor are
the only therapeutically active ingredients for treating a cancer.
Additional combinations of three or more agents are encompassed by
the methods and compositions described herein.
[0062] Additional combinations of three or more agents are
encompassed by the methods and compositions described herein.
[0063] In some embodiments of the compositions and methods
described herein, the PI3K inhibitor and the Bcl-2 inhibitor are in
a single dosage form. In other embodiments, the PI3K inhibitor and
the Bcl-2 inhibitor are in separate dosage forms.
[0064] In some embodiments of the compositions and methods
described herein, the combination of the PI3K inhibitor and the
Bcl-2 inhibitor is synergistic, e.g., in inhibiting tumor cell
growth, viability or both, or in treating a cancer.
[0065] In certain embodiments, the Bcl-2 inhibitor is ABT-199,
ABT-263, ABT-737, G3139 (genasense or oblimersen), GX15-070
(obatoclax mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101),
antimycin A, apogossypol, S44563, or a combination or mixture
thereof. In one embodiment, the Bcl-2 inhibitor is ABT-199. In one
embodiment, the Bcl-2 inhibitor is ABT-263.
[0066] In certain embodiments, provided herein is a composition
(e.g., a pharmaceutical composition) comprising a PI3K inhibitor
(e.g., Compound 1 or GS1101), or a pharmaceutically acceptable form
thereof, in combination with ABT-199, or a pharmaceutically
acceptable form thereof. The PI3K inhibitor and ABT-199 can be
present in a single composition or as two or more different
compositions. In some embodiments, the composition (e.g., one or
more compositions comprising the combination of PI3K inhibitor and
ABT-199) is synergistic, e.g., has a synergistic effect in treating
a cancer (e.g., in reducing cancer cell growth or viability, or
both, e.g., as described herein). In certain embodiments, the
amount or dosage of the PI3K inhibitor, ABT-199, or both, present
in the composition(s) is lower (e.g., at least 20%, at least 30%,
at least 40%, or at least 50% lower) than the amount or dosage of
each agent used individually, e.g., as a monotherapy.
[0067] In certain embodiments, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject a PI3K inhibitor,
e.g., one or more PI3K inhibitors (e.g., Compound 1 or GS1101, or
both) or a pharmaceutically acceptable form thereof, in combination
with ABT-199, or a pharmaceutically acceptable form thereof. In
certain embodiments, the combination of the PI3K inhibitor and
ABT-199 is synergistic, e.g., has a synergistic effect in treating
the cancer (e.g., in reducing cancer cell growth or viability, or
both). In some embodiments, the amount or dosage of the PI3K
inhibitor, ABT-199, or both, used in combination does not exceed
the level at which each agent is used individually, e.g., as a
monotherapy. In certain embodiments, the amount or dosage of the
PI3K inhibitor, ABT-199, or both, used in combination is lower
(e.g., at least 20%, at least 30%, at least 40%, or at least 50%
lower) than the amount or dosage of each agent used individually,
e.g., as a monotherapy. In other embodiments, the amount or dosage
of the PI3K inhibitor, ABT-199, or both, used in combination that
results in treatment of cancer is lower (e.g., at least 20%, at
least 30%, at least 40%, or at least 50% lower) than the amount or
dosage of each agent used individually, e.g., as a monotherapy.
[0068] In certain embodiments, provided herein is a composition
(e.g., a pharmaceutical composition) comprising a PI3K inhibitor
(e.g., Compound 1 or GS1101), or a pharmaceutically acceptable form
thereof, in combination with ABT-263, or a pharmaceutically
acceptable form thereof. The PI3K inhibitor and ABT-263 can be
present in a single composition or as two or more different
compositions. In some embodiments, the composition (e.g., one or
more compositions comprising the combination of PI3K inhibitor and
ABT-263) is synergistic, e.g., has a synergistic effect in treating
a cancer (e.g., in reducing cancer cell growth or viability, or
both, e.g., as described herein). In certain embodiments, the
amount or dosage of the PI3K inhibitor, ABT-263, or both, present
in the composition(s) is lower (e.g., at least 20%, at least 30%,
at least 40%, or at least 50% lower) than the amount or dosage of
each agent used individually, e.g., as a monotherapy.
[0069] In certain embodiments, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject a PI3K inhibitor,
e.g., one or more PI3K inhibitors or a pharmaceutically acceptable
form thereof, in combination with ABT-263, or a pharmaceutically
acceptable form thereof. In certain embodiments, the combination of
the PI3K inhibitor and ABT-263 is synergistic, e.g., has a
synergistic effect in treating the cancer (e.g., in reducing cancer
cell growth or viability, or both). In some embodiments, the amount
or dosage of the PI3K inhibitor, ABT-263, or both, used in
combination does not exceed the level at which each agent is used
individually, e.g., as a monotherapy. In certain embodiments, the
amount or dosage of the PI3K inhibitor, ABT-263, or both, used in
combination is lower (e.g., at least 20%, at least 30%, at least
40%, or at least 50% lower) than the amount or dosage of each agent
used individually, e.g., as a monotherapy. In other embodiments,
the amount or dosage of the PI3K inhibitor, ABT-263, or both, used
in combination that results in treatment of cancer is lower (e.g.,
at least 20%, at least 30%, at least 40%, or at least 50% lower)
than the amount or dosage of each agent used individually, e.g., as
a monotherapy.
[0070] In one embodiment, the PI3K inhibitor is chosen from one or
more of: Compound 1, AMG-319, GSK 2126458, GSK 1059615, GDC-0032,
GDC-0980, GDC-0941, XL147, XL499, XL765, BKM 120 GS1101, CAL 263,
SF1126, PX-866, BEZ235, CAL-120, BYL719, RP6503, RP6530, TGR1202,
INK1117, PX-886, BAY 80-6946, IC87114, Palomid 529, ZSTK474,
PWT33597, TG100-115, GNE-477, CUDC-907, AEZS-136, BGT-226,
PF-05212384, LY3023414, PI-103, LY294002, INCB-040093, CAL-130 or
wortmannin.
[0071] Embodiments relating to dosages of the agents included in
the compositions and methods described herein follow. In one
embodiment, the PI3K inhibitor, e.g., Compound 1, is administered
at a dosage of from about 0.01 mg to about 75 mg daily, and the
Bcl-2 inhibitor is administered at a dosage of from about 0.01 to
about 1100 mg daily.
[0072] In certain embodiments, the amount or dosage of the PI3K
inhibitor, the Bcl-2 inhibitor, or both, that is used in the method
or composition is lower (e.g., at least 20%, at least 30%, at least
40%, at least 50%, at least 60%, at least 70%, or at least 80%
lower) than the amount or dosage of each agent used individually,
e.g., as a monotherapy. In other embodiments, the amount or dosage
of the PI3K inhibitor, the Bcl-2 inhibitor, or both, present in the
composition(s) that results in a desired effect (e.g., treatment of
cancer) is lower (e.g., at least 20%, at least 30%, at least 40%,
at least 50%, at least 60%, at least 70%, or at least 80% lower)
than the amount or dosage of each agent used individually, e.g., as
a monotherapy.
[0073] In one embodiment, the composition comprises the PI3K
inhibitor, or a pharmaceutically acceptable form thereof, at an
amount of in the range of from about 0.01 mg to about 75 mg.
[0074] In one embodiment, the molar ratio of Compound 1, or a
pharmaceutically acceptable form thereof, to the Bcl-2 inhibitor,
or a pharmaceutically acceptable form thereof, is in the range of
from about 50:1 to about 1:50. In one embodiment, the molar ratio
of Compound 1, or a pharmaceutically acceptable form thereof, to
the Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
is in the range of from about 10:1 to about 1:10 or from about 1:3
to about 1:7.
[0075] In one embodiment, the composition comprises Compound 1, or
a pharmaceutically acceptable form thereof, at an amount in the
range of from about 0.01 mg to about 75 mg and the Bcl-2 inhibitor,
or a pharmaceutically acceptable form thereof, at an amount of in
the range of from about 0.01 mg to about 1100 mg. In one
embodiment, the composition comprises ABT-199 at an amount of about
400 mg, about 325 mg, about 150 mg, or about 75 mg.
[0076] In some embodiments, the PI3K inhibitor is Compound 1 at a
dosage of 25 mg (e.g., 25 mg BID). In certain embodiments, Compound
1 is effective as a monotherapy at a dosage of 25 mg (e.g., 25 mg
BID). In certain embodiments, the combination of Compound 1 and the
Bcl-2 inhibitor is effective, e.g., in treating a cancer and/or in
reducing cancer cell growth or viability, with Compound 1 at a
dosage lower than 25 mg (e.g., 25 mg BID). In certain embodiments,
the dosage of Compound 1 is 22.5 mg (e.g., 22.5 mg BID), 20 mg
(e.g., 20 mg BID), 17.5 mg (e.g., 17.5 mg BID), 15 mg (e.g., 15 mg
BID), 12.5 mg (e.g., 12.5 mg BID), 10 mg (e.g., 10 mg BID), 7.5 mg
(e.g., 7.5 mg BID), or 5 mg (e.g., 5 mg BID).
[0077] In some embodiments, the PI3K inhibitor, e.g., Compound 1,
is administered at a dose frequency of twice per day (BID), once
per day, once per two days, once per three days, once per four
days, once per five days, once per six days, or once per week. In
certain embodiments, the combination of the PI3K inhibitor (e.g.,
Compound 1) and the Bcl-2 inhibitor is effective, e.g., in treating
a cancer and/or in reducing cancer cell growth or viability, with
the PI3K inhibitor (e.g., Compound 1) administered at a dose
frequency of twice per day (BID), once per day, once per two days,
once per three days, once per four days, once per five days, once
per six days, or once per week.
[0078] In some embodiments, the PI3K inhibitor is GS1101 at a
dosage of 150 mg (e.g., 150 mg BID). In certain embodiments, GS1101
is effective as a monotherapy at a dosage of 150 mg (e.g., 150 mg
BID). In certain embodiments, the combination of GS1101 and the
Bcl-2 inhibitor is effective, e.g., in treating a cancer and/or in
reducing cancer cell growth or viability, with GS1101 at a dosage
lower than 150 mg (e.g., 150 mg BID). In certain embodiments, the
dosage of GS1101 is 135 mg (e.g., 135 mg BID), 120 mg (e.g., 120 mg
BID), 105 mg (e.g., 105 mg BID), 90 mg (e.g., 90 mg BID), 75 mg
(e.g., 75 mg BID), 60 mg (e.g., 60 mg BID), 45 mg (e.g., 45 mg
BID), or 30 mg (e.g., 30 mg BID).
[0079] In some embodiments, the PI3K inhibitor is GS1101 and is
administered at a dose frequency of twice per day, once per day,
once per two days, once per three days, once per four days, once
per five days, once per six days, or once per week. In certain
embodiments, the combination of GS1101 and the Bcl-2 inhibitor is
effective, e.g., in treating a cancer and/or in reducing cancer
cell growth or viability, with GS1101 administered at a dose
frequency of twice per day (BID), once per day, once per two days,
once per three days, once per four days, once per five days, once
per six days, or once per week.
[0080] In some embodiments, the Bcl-2 inhibitor is ABT-199 at a
dosage of 400 mg (e.g., 400 mg QID). In certain embodiments,
ABT-199 is effective as a monotherapy at a dosage of 400 mg (e.g.,
400 mg QID). In certain embodiments, the combination of the PI3K
inhibitor and ABT-199 is effective, e.g., in treating a cancer
and/or in reducing cancer cell growth or viability, with ABT-199 at
a dosage lower than 400 mg (e.g., 400 mg QID). In certain
embodiments, the dosage of ABT-199 is 50 to 350 mg (e.g., 50 to 350
mg BID), e.g., 50 to 100 mg (e.g., 50 to 100 mg BID), 100 to 200 mg
(e.g., 100 to 200 mg BID), 200 to 300 mg (e.g., 200 to 300 mg BID)
or 250 to 350 mg (e.g., 250 to 350 mg BID). In certain embodiments,
the dosage of ABT-199 is about 350 mg (e.g., 350 mg QID), 300 mg
(e.g., 300 mg QID), 250 mg (e.g., 250 mg QID), 200 mg (e.g., 200 mg
QID), 150 mg (e.g., 150 mg QID), 100 mg (e.g., 100 mg QID), or 50
mg (e.g., 50 mg QID).
[0081] In some embodiments, the Bcl-2 inhibitor is a Bcl-2
inhibitor (e.g., ABT-199) administered at a dose frequency of four
times per day (QID), three times per day, twice per day (BID), once
per day, once per two days, once per three days, once per four
days, once per five days, once per six days, or once per week. In
certain embodiments, the combination of the PI3K inhibitor and
Bcl-2 inhibitor (e.g., ABT-199) is effective, e.g., in treating a
cancer and/or in reducing cancer cell growth or viability, when the
Bcl-2 inhibitor (e.g., ABT-199) administered at a dose frequency of
four times per day (QID), three times per day, twice per day (BID),
once per day, once per two days, once per three days, once per four
days, once per five days, once per six days, or once per week.
[0082] In some embodiments, the Bcl-2 inhibitor is ABT-263 at a
dosage of 325 mg. In certain embodiments, ABT-263 is effective as a
monotherapy at a dosage of 325 mg. In certain embodiments, the
combination of the PI3K inhibitor and ABT-263 is effective, e.g.,
in treating a cancer and/or in reducing cancer cell growth or
viability, with ABT-263 at a dosage lower than 325 mg. In certain
embodiments, the dosage of ABT-263 is 50 to 300 mg (e.g., 50 to 100
mg, 100 to 200 mg, or 200 to 300 mg). In certain embodiments, the
dosage of ABT-263 is about 300 mg, 275 mg, 250 mg, 225 mg, 200 mg,
175 mg, 150 mg, 125 mg, 100 mg, 75 mg, or 50 mg. In certain
embodiments, the dosage is for daily administration.
[0083] In some embodiments, the Bcl-2 inhibitor is ABT-263 and is
administered at a dose frequency of four times per day (QID), three
times per day, twice per day (BID), once per day, once per two
days, once per three days, once per four days, once per five days,
once per six days, or once per week. In certain embodiments, the
combination of the PI3K inhibitor and ABT-263 is effective, e.g.,
in treating a cancer and/or in reducing cancer cell growth or
viability, when ABT-263 administered at a dose frequency of four
times per day (QID), three times per day, twice per day (BID), once
per day, once per two days, once per three days, once per four
days, once per five days, once per six days, or once per week.
[0084] In one embodiment, the Bcl-2 inhibitor is administered to a
subject at least 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, 12 weeks, or 16 weeks before the PI3K inhibitor
(e.g., Compound 1), or a pharmaceutically acceptable form thereof,
is administered. In another embodiment, the Bcl-2 inhibitor is
administered concurrently with the PI3K inhibitor (e.g., Compound
1), or a pharmaceutically acceptable form thereof, e.g., in a
single dosage form or separate dosage forms. In yet another
embodiment, the Bcl-2 inhibitor is administered to the subject at
least 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, 12 weeks, or 16 weeks after the PI3K inhibitor (e.g.,
Compound 1), or a pharmaceutically acceptable form thereof, is
administered.
[0085] In one embodiment, Compound 1, or a pharmaceutically
acceptable form thereof, and the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, are the
only therapeutically active ingredients.
[0086] In one embodiment, Compound 1, or a pharmaceutically
acceptable form thereof, and the Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, are in a single dosage
form. In one embodiment, Compound 1, or a pharmaceutically
acceptable form thereof, and the Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, are in separate dosage
forms. In one embodiment, the molar ratio of Compound 1, or a
pharmaceutically acceptable form thereof, to the Bcl-2 inhibitor,
or a pharmaceutically acceptable form thereof, is in the range of
from about 50:1 to about 1:50. In one embodiment, the molar ratio
of Compound 1, or a pharmaceutically acceptable form thereof, to
the Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
is in the range of from about 10:1 to about 1:10 or from about 1:3
to about 1:7.
[0087] In one embodiment, Compound 1 is administered at an amount
to reach maximum plasma concentration at steady state (Cmaxss) at
about 1000 ng/mL to about 5000 ng/mL; and the Bcl-2 inhibitor is
administered at an amount to reach Cmaxss at about 0.1 .mu.g/mL to
about 1000 .mu.g/mL.
[0088] In one embodiment, Compound 1 is administered at an amount
to reach an area under the plasma concentration-time curve at
steady-state (AUCss) at about 5000 ng/mL*hr to about 10000
ng/mL*hr; and the Bcl-2 inhibitor is administered at an amount to
reach an AUCss at about 0.1 ng/mL*hr to about 10000 ng/mL*hr.
[0089] In some embodiments, the PI3K inhibitor is Compound 1 and is
effective (e.g., therapeutically effective) in the combination at
an amount that is decreased by about 1.5 fold to about 50 fold of
the amount that is required to achieve the same effect, e.g., the
same therapeutic effect, when Compound 1 is administered as a
monotherapy. In certain embodiments, the Bcl-2 inhibitor is
effective (e.g., therapeutically effective) in the combination at
an amount that is decreased by about 1.5 fold to about 50 fold of
the amount that is required to achieve the same effect, e.g., the
same therapeutic effect, when Compound 1 is administered as a
monotherapy.
[0090] In some embodiments, the PI3K inhibitor is Compound 1, the
Bcl-2 inhibitor is ABT-199 or ABT-263, and the cancer is a diffuse
large B-cell lymphoma. In some such embodiments, the molar ratio of
Compound 1, or a pharmaceutically acceptable form thereof, to the
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof, is
in the range of from about 50:1 to about 1:50.
[0091] In some embodiments, the PI3K inhibitor is Compound 1, the
Bcl-2 inhibitor is ABT-199 or ABT-263, and the cancer is a
follicular lymphoma. In some such embodiments, the molar ratio of
Compound 1, or a pharmaceutically acceptable form thereof, to the
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof, is
in the range of from about 50:1 to about 1:50.
[0092] In some embodiments, the PI3K inhibitor is Compound 1, the
Bcl-2 inhibitor is ABT-199 or ABT-263, and the cancer is a T-cell
lymphoma. In some such embodiments, the molar ratio of Compound 1,
or a pharmaceutically acceptable form thereof, to the Bcl-2
inhibitor, or a pharmaceutically acceptable form thereof, is in the
range of from about 50:1 to about 1:50.
[0093] In some embodiments, the PI3K inhibitor is Compound 1, the
Bcl-2 inhibitor is ABT-199 or ABT-263, and the cancer is mantle
cell lymphoma. In some such embodiments, the molar ratio of
Compound 1, or a pharmaceutically acceptable form thereof, to the
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof, is
in the range of from about 50:1 to about 1:50.
[0094] In one embodiment, provided herein is a method of reducing
or delaying resistance to a treatment with PI3K inhibitor (e.g., a
treatment with one or more PI3K inhibitors described herein) in a
subject, or a method of reducing the likelihood for a subject to
develop resistance to a treatment with a PI3K inhibitor (e.g., a
treatment with one or more PI3K inhibitors described herein), the
method comprising:
[0095] (a) administering to the subject a therapeutically effective
amount of a monotherapy comprising the PI3K inhibitor, or a
pharmaceutically acceptable form thereof, for a first period of
time;
[0096] (b) after the first period of time, administering to the
subject a therapeutically effective amount of a combination therapy
comprising the PI3K inhibitor in combination with a Bcl-2 inhibitor
(e.g., one or more Bcl-2 inhibitors as described herein), for a
second period of time; and
[0097] (c) optionally repeating steps (a) and (b) one or more
times.
[0098] In one embodiment, provided herein is a method of reducing
or delaying resistance to a treatment with PI3K inhibitor (e.g., a
treatment with one or more PI3K inhibitors described herein) in a
subject, or a method of reducing the likelihood for a subject to
develop resistance to a treatment with a PI3K inhibitor (e.g., a
treatment with one or more PI3K inhibitors described herein), the
method comprising:
[0099] (a) administering to the subject a therapeutically effective
amount of a monotherapy comprising the Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, for a first period of
time;
[0100] (b) after the first period of time, administering to the
subject a therapeutically effective amount of a combination therapy
comprising a PI3K inhibitor in combination with the Bcl-2 inhibitor
(e.g., one or more Bcl-2 inhibitors as described herein), for a
second period of time; and
[0101] (c) optionally repeating steps (a) and (b) one or more
times.
[0102] In certain embodiments, the subject is identified as
developing resistance (e.g., acquired resistance) to the
monotherapy. In certain embodiments, the subject is identified as
developing resistance based on an assessment of MRD. In certain
embodiments, the subject is identified as developing resistance if
the MRD is above a preselected cutoff value (e.g., 1 malignant cell
in 100 normal cells, 1 malignant cell in 1000 normal cells, or 1
malignant cell in 10,000 normal cells). In certain embodiments, the
preselected cutoff value is 1 malignant cell in 1000 or 10,000
normal cells. In certain embodiments, the method further comprises
assessing MRD, e.g., in step (a) and/or (b).
[0103] In certain aspects, this disclosure provides a method of
delaying resistance of a subject having a cancer, comprising
administering to the subject a synergistic amount of a PI3K
inhibitor, or a pharmaceutically acceptable form thereof, and a
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
thereby delaying resistance (e.g., resistance to the PI3K
inhibitor). The method may comprise administering the PI3K
inhibitor before the Bcl-2 inhibitor.
[0104] The disclosure also provides, in certain aspects, a method
of reducing the risk that a cancer becomes resistant to the PI3K
inhibitor, comprising administering to a subject having a cancer a
synergistic amount of a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, and a Bcl-2 inhibitor, or a
pharmaceutically acceptable form thereof, thereby reducing the risk
that the cancer becomes resistant to the PI3K inhibitor.
[0105] The disclosure also provides, in certain aspects, a method
of prolonging remission (e.g., complete remission or partial
remission) in a subject having a cancer, comprising administering
to the subject a synergistic amount of a PI3K inhibitor, or a
pharmaceutically acceptable form thereof, and a Bcl-2 inhibitor, or
a pharmaceutically acceptable form thereof, thereby prolonging
remission in the subject.
[0106] The disclosure also provides, in certain aspects, a method
of increasing the likelihood that a subject having a cancer
experiences complete remission, comprising administering to the
subject a synergistic amount of a PI3K inhibitor, or a
pharmaceutically acceptable form thereof, and a Bcl-2 inhibitor, or
a pharmaceutically acceptable form thereof, thereby increasing the
likelihood that the subject experiences complete remission.
[0107] The disclosure also provides, in certain aspects, a method
of reducing the level of minimal residual disease (MRD) compared to
a reference value (e.g., compared to a pre-treatment value or a
value obtained during treatment) in a subject having a cancer,
comprising administering to the subject a synergistic amount of a
PI3K inhibitor, or a pharmaceutically acceptable form thereof, and
a Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
thereby reducing the level of MRD in the subject.
[0108] This disclosure further provides methods of assessing a
subject's cells, and treating the patient with a combination of a
PI3K inhibitor and a Bcl-2 inhibitor.
[0109] For instance, in certain aspects, the invention features a
method of evaluating the responsiveness of a cancer or tumor, or a
subject having a cancer or tumor, to a treatment with a BCR pathway
inhibitor (e.g., a treatment with an inhibitor of PI3K, BTK or SYK,
alone or in combination). In one embodiment, responsiveness to a
PI3K inhibitor is evaluated. The method includes: acquiring a value
(e.g., determining one or more of: the presence, absence, amount or
level) of an alteration or biomarker chosen from one, two, three,
four or all of: an STK11 copy number, TSC1 copy number, TSC2 copy
number, a p53 pathway mutation (e.g., a mutation disclosed in Table
20), or MAPK pathway mutation (e.g., a mutation disclosed in Table
18), or any combination thereof (e.g., a dual MAPK/p53 pathway
mutation, e.g., a mutation disclosed in Table 18 and a mutation
disclosed in Table 20). The method further comprises administering
a PI3K inhibitor and a Bcl-2 inhibitor to the subject. In some
embodiments, the subject has, or is identified as having, a tumor
that is responsive to the PI3K inhibitor. In some embodiments, the
subject has, or is identified as having, a tumor that is not
responsive to the PI3K inhibitor. In some embodiments, PI3K
inhibitor and a Bcl-2 inhibitor are administered to a subject that
has, or is identified as having, a tumor that is less responsive,
e.g., not responsive to the PI3K inhibitor. In some embodiments,
PI3K inhibitor and a Bcl-2 inhibitor are administered to a subject
that has, or is identified as having, a tumor that is responsive to
the PI3K inhibitor. In some embodiments, PI3K inhibitor and a Bcl-2
inhibitor are administered to a subject that has, or is identified
as having, a tumor that is less responsive, e.g., not responsive to
the PI3K inhibitor. In some embodiments, co-administration of the
Bcl-2 inhibitor improves sensitivity of the tumor to the PI3K
inhibitor.
[0110] In another aspect, the invention features a method of
monitoring a treatment of a subject with a BCR pathway inhibitor
(e.g., a treatment with an inhibitor of PI3K, BTK or SYK, alone or
in combination). In one embodiment, treatment with a PI3K inhibitor
is monitored. The method includes: acquiring, at two or more time
intervals, a value (e.g., determining one or more of: the presence,
absence, amount or level) of an alteration or biomarker chosen from
one, two, three, four or all of: an STK11 copy number, TSC1 copy
number, TSC2 copy number, a p53 pathway mutation (e.g., a mutation
disclosed in Table 20), or MAPK pathway mutation (e.g., a mutation
disclosed in Table 18), or any combination thereof (e.g., a dual
MAPK/p53 mutation, e.g., a mutation disclosed in Table 18 and a
mutation disclosed in Table 20). The method further comprises
administering a PI3K inhibitor and a Bcl-2 inhibitor to the
subject.
[0111] In another aspect, the invention features a method of
treating (e.g., inhibiting, reducing, ameliorating, managing, or
preventing) a cancer or tumor in a subject. The method includes:
acquiring a value (e.g., determining one or more of: the presence,
absence, amount or level) of an alteration or biomarker chosen from
one, two, three, four or all of: an STK11 copy number, TSC1 copy
number, TSC2 copy number, a p53 pathway mutation (e.g., a mutation
disclosed in Table 20), or MAPK pathway mutation (e.g., a mutation
disclosed in Table 18), or any combination thereof (e.g., a dual
MAPK/p53 mutation, e.g., a mutation disclosed in Table 18 and a
mutation disclosed in Table 20), and responsive to said value,
administering to the subject a BCR pathway inhibitor, e.g., a PI3K
inhibitor (e.g., one or more PI3K inhibitors) and a Bcl-2
inhibitor.
[0112] In another aspect, the invention features a method of
treating a subject, comprising (i) administering a first treatment
comprising a first PI3K inhibitor to the subject (ii) acquiring
information regarding the presence or absence of an alteration in a
biomarker in one or more samples from the subject, wherein the
biomarker is selected from STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3,
YWHAE, PER1, GAS7, FSTL3, USP6, MAP2K4, or EGFR; and (iii)
continuing administration of the first treatment if the alteration
is absent, or administering a second treatment if the alteration is
present, wherein the second treatment includes administration of a
Bcl-2 inhibitor.
[0113] In certain embodiments, the alteration is an STK11, TSC1,
TSC2, TP53, PTEN, CBFA2T3, YWHAE, PER1, GAS7, FSTL3, USP6, or
MAP2K4 copy number loss (e.g., single copy loss). In some
embodiments, the STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3, YWHAE,
PER1, GAS7, FSTL3, USP6, or MAP2K4 copy number in a sample taken
from the subject after the first treatment is lower than a
corresponding STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3, YWHAE, PER1,
GAS7, FSTL3, USP6, MAP2K4 copy number in a sample taken from the
subject before the first treatment (e.g., there is an STK11 single
copy loss).
[0114] In another aspect, the present disclosure provides a method
of evaluating the responsiveness of a cancer or tumor, or a subject
having a cancer or tumor, to a treatment with a BCR pathway
inhibitor (e.g., a treatment with an inhibitor of PI3K, BTK or SYK,
alone or in combination). In one embodiment, responsiveness to a
PI3K inhibitor is evaluated. The method includes: acquiring a value
(e.g., determining one or more of: the presence, absence, amount or
level) of an anti-apoptotic factor such as Bcl-2.
[0115] In another aspect, the invention features a method of
monitoring a treatment of a subject with a BCR pathway inhibitor
(e.g., a treatment with an inhibitor of PI3K, BTK or SYK, alone or
in combination). In one embodiment, treatment with a PI3K inhibitor
is monitored. The method includes: acquiring, at two or more time
intervals, a value (e.g., determining one or more of: the presence,
absence, amount or level) of an anti-apoptotic factor such as
Bcl-2.
[0116] In another aspect, the invention features a method of
treating (e.g., inhibiting, reducing, ameliorating, managing, or
preventing) a cancer or tumor in a subject. The method includes:
acquiring a value (e.g., determining one or more of: the presence,
absence, amount or level) of an anti-apoptotic factor such as
Bcl-2.
[0117] In certain embodiments, the methods that include acquiring a
value of Bcl-2 also include acquiring a value (e.g., determining
one or more of: the presence, absence, amount or level) of a
pro-apoptotic factor or anti-apoptotic factor. The pro-apoptotic
factor can be, e.g., one or more of (e.g., 2, 3, 4, or all of) BMF,
BIK, BIM, NOXA, PUMA, and HRK. In some embodiments, an elevated
level of Bch 2 indicates that the cancer is resistant to a PI3K
inhibitor. In some embodiments, a normal or reduced level of Bcl-2
indicates that the cancer is responsive to a PI3K inhibitor. In
some embodiments, an elevated level of one or more pro-apoptotic
factors (e.g., BMF, BIK, BIM, NOXA, PUMA, and HRK) indicates that
the cancer is more responsive to a PI3K inhibitor (optionally in
combination with a Bcl-2 inhibitor) than a cancer with normal or
lowered levels of the pro-apoptotic factor. In some embodiments,
the methods involve administering a Bcl-2 inhibitor (e.g., in
combination with a PI3K inhibitor) to a subject having elevated
Bcl-2 levels. In some embodiments, the methods involve
administering a PI3K inhibitor as a monotherapy to a subject having
normal or low Bcl-2 levels. In some embodiments, the elevated,
normal, or reduced levels of a biomarker are determined with
reference to a non-cancerous control value.
[0118] The disclosure includes all combinations of any one or more
of the foregoing aspects and/or embodiments, as well as
combinations with any one or more of the embodiments set forth in
the detailed description and examples.
INCORPORATION BY REFERENCE
[0119] All publications, patents, and patent applications mentioned
in this specification are herein incorporated by reference in their
entirety and to the same extent as if each individual publication,
patent, or patent application is specifically and individually
indicated to be incorporated by reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0120] FIG. 1 shows an isobologram depicting the synergistic effect
of the combination of Compound 1 and ABT-199 in SU-DHL-4 cell
line.
[0121] FIG. 2 shows an isobologram depicting the synergistic effect
of the combination of Compound 1 and ABT-199 in Mino cell line.
[0122] FIG. 3 shows an isobologram depicting the synergistic effect
of the combination of Compound 1 and ABT-199 in WSU-NHL cell
line.
[0123] FIG. 4 shows an isobologram depicting the synergistic effect
of the combination of Compound 1 and ABT-199 in DoHH-2 cell
line.
[0124] FIGS. 5A, 5B, and 5C show the change in RNA levels of
selected differentially regulated genes in patients treated with
Compound 1.
[0125] FIG. 6A is a graphical representation of the change in RNA
levels of selected genes in patients treated with Compound 1 after
8 days.
[0126] FIG. 6B is a graphical representation of the change in RNA
levels of selected genes in patients treated with Compound 1 after
28 days.
[0127] FIG. 7 is a graph showing the fold change in Harakiri
expression level in partial remission and stable disease patients
after 7 days of Compound 1 treatment.
[0128] FIG. 8 is a graphical representation of the relationship
between mutations and responses to Compound 1. Each column
represents a patient. Each row represents a mutation. The diagnosis
is coded as 1: CLL/SLL (R/R), or 2: CLL/SLL (treatment-naive). R/R
refers to a patient that has relapsed or is refractory to
treatment. Tx naive refers to a patient that is treatment naive,
e.g., has not been previously administered Compound 1. The response
is coded as 3: CR/PR, 4: PRwL, 5: SD/PD, or 6: SD/PD (nodal
response). The ALC is coded as 7: high, 8: normal, or 9: low. PR
refers to partial remission, SD refers to stable disease, PD refers
to progressive disease, and CR refers to complete remission
[0129] FIG. 9 is a graphical representation of the relationship
between mutations and responses to Compound 1. Each column
represents a patient. Each row represents a mutation. The diagnosis
is coded as 1: CLL/SLL (R/R), or 2: CLL/SLL (treatment-naive). The
response is coded as IWCLL complete remission or partial remission
(CR/PR) or IWCLL stable disease or progressive disease (SD/PD).
Nodal responses are indicated with an asterisk (*).
[0130] FIG. 10 is a graphical representation of the relationship
between mutations and responses to Compound 1. The diagnosis and
response is coded as in FIG. 9.
[0131] FIG. 11 is a graphical representation of the relationship
between mutations and responses to Compound 1. The diagnosis and
response is coded as in FIG. 9. Nodal responses are indicated with
an asterisk (*). A non-assessable nodal response is indicated by a
(#).
[0132] FIG. 12 is a graphical representation of the relationship
between CLL common copy number variations (CNVs) and responses to
Compound 1. The diagnosis and response is coded as in FIG. 10.
[0133] FIG. 13A is a graph depicting relative expression of TP53
(RNA levels) in patients with no loss or with a loss in TP53 copy
number. FIG. 13B is a graph depicting relative expression of YWHAE
(RNA levels) in patients with no loss or with a loss in YWHAE copy
number. FIG. 13C is a graph depicting relative expression of STK11
(RNA levels) in patients with no loss or with a loss in STK11 copy
number.
[0134] FIG. 14 is a graphical representation of the relationship
between and responses to Compound 1 and alterations in various
pathways. "Dual" in this figure refers to dual p53 and MAPK
pathways. The diagnosis and response is coded as in FIG. 9.
[0135] FIG. 15 is a graph showing the IC50 for Compound 1
inhibition of several cell clones.
[0136] FIG. 16 is a graph showing the PTEN RNA expression level in
DMSO control treated cells or cells resistant to Compound 1. FPKM
refers to fragments per kilobase of exon per million fragments
mapped.
DETAILED DESCRIPTION
1. Definitions
[0137] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as is commonly understood by one
of skill in the art to which this specification pertains.
[0138] As used in the specification and claims, the singular form
"a", "an" and "the" includes plural references unless the context
clearly dictates otherwise.
[0139] As used herein, and unless otherwise indicated, the term
"about" or "approximately" means an acceptable error for a
particular value as determined by one of ordinary skill in the art,
which depends in part on how the value is measured or determined.
In certain embodiments, the term "about" or "approximately" means
within 1, 2, 3, or 4 standard deviations. In certain embodiments,
the term "about" or "approximately" means within 50%, 20%, 15%,
10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given
value or range.
[0140] The term "agonist" as used herein refers to a compound or
agent having the ability to initiate or enhance a biological
function of a target protein or polypeptide, such as increasing the
activity or expression of the target protein or polypeptide.
Accordingly, the term "agonist" is defined in the context of the
biological role of the target protein or polypeptide. While some
agonists herein specifically interact with (e.g., bind to) the
target, compounds and/or agents that initiate or enhance a
biological activity of the target protein or polypeptide by
interacting with other members of the signal transduction pathway
of which the target polypeptide is a member are also specifically
included within this definition.
[0141] The terms "antagonist" and "inhibitor" are used
interchangeably, and they refer to a compound or agent having the
ability to reduce or inhibit a biological function of a target
protein or polypeptide, such as by reducing or inhibiting the
activity or expression of the target protein or polypeptide.
Accordingly, the terms "antagonist" and "inhibitor" are defined in
the context of the biological role of the target protein or
polypeptide. An inhibitor need not completely abrogate the
biological function of a target protein or polypeptide, and in some
embodiments reduces the activity by at least 50%, 60%, 70%, 80%,
90%, 95%, or 99%. While some antagonists herein specifically
interact with (e.g., bind to) the target, compounds that inhibit a
biological activity of the target protein or polypeptide by
interacting with other members of the signal transduction pathway
of which the target protein or polypeptide are also specifically
included within this definition. Non-limiting examples of
biological activity inhibited by an antagonist include those
associated with the development, growth, or spread of a tumor, or
an undesired immune response as manifested in autoimmune
disease.
[0142] The term "effective amount" or "therapeutically effective
amount" refers to that amount of a compound or pharmaceutical
composition described herein that is sufficient to effect the
intended application including, but not limited to, disease
treatment, as illustrated below. The therapeutically effective
amount can vary depending upon the intended application (in vitro
or in vivo), or the subject and disease condition being treated,
e.g., the weight and age of the subject, the severity of the
disease condition, the manner of administration and the like, which
can readily be determined by one of ordinary skill in the art. The
term also applies to a dose that will induce a particular response
in target cells, e.g., reduction of platelet adhesion and/or cell
migration. The specific dose will vary depending on, for example,
the particular compounds chosen, the dosing regimen to be followed,
whether it is administered in combination with other agents, timing
of administration, the tissue to which it is administered, and the
physical delivery system in which it is carried.
[0143] As used herein, a daily dosage can be achieved by a single
administration of the targeted dosage amount or multiple
administrations of smaller dosage amount(s). For example, a 150 mg
daily dosage can be achieved by a single administration of 150 mg
of the therapeutic agent per day, two administrations of 75 mg of
the therapeutic agent per day, or three administrations of 50 mg of
the therapeutic agent per day, or the like.
[0144] As used herein, the terms "treatment", "treating",
"palliating" and "ameliorating" are used interchangeably herein.
These terms refer to an approach for obtaining beneficial or
desired results including, but not limited to, therapeutic benefit.
By therapeutic benefit is meant eradication or amelioration of the
underlying disorder being treated. Also, a therapeutic benefit is
achieved with the eradication or amelioration of one or more of the
physiological symptoms associated with the underlying disorder such
that an improvement is observed in the patient, notwithstanding
that the patient can still be afflicted with the underlying
disorder.
[0145] As used herein, the terms "prevention" and "preventing" are
used herein to refer to an approach for obtaining beneficial or
desired results including, but not limited, to prophylactic
benefit. For prophylactic benefit, the pharmaceutical compositions
may be administered to a patient at risk of developing a particular
disease, or to a patient reporting one or more of the physiological
symptoms of a disease, even though a diagnosis of this disease may
not have been made.
[0146] A "therapeutic effect," as that term is used herein,
encompasses a therapeutic benefit and/or a prophylactic benefit as
described above. A prophylactic effect includes delaying or
eliminating the appearance of a disease or condition, delaying or
eliminating the onset of symptoms of a disease or condition,
slowing, halting, or reversing the progression of a disease or
condition, or any combination thereof.
[0147] The phrase "a method of treating" or its equivalent, when
applied to, for example, cancer refers to a procedure or course of
action that is designed to reduce or eliminate the number of cancer
cells in an animal, or to alleviate the symptoms of a cancer. "A
method of treating" cancer or another proliferative disorder does
not necessarily mean that the cancer cells or other disorder will,
in fact, be eliminated, that the number of cells or disorder will,
in fact, be reduced, or that the symptoms of a cancer or other
disorder will, in fact, be alleviated. Often, a method of treating
cancer will be performed even with a low likelihood of success, but
which, given the medical history and estimated survival expectancy
of an animal, is nevertheless deemed an overall beneficial course
of action.
[0148] The term "therapeutically effective agent" means a
composition that will elicit the biological or medical response of
a tissue, system, animal or human that is being sought by the
researcher, veterinarian, medical doctor or other clinician.
[0149] As used herein, the "aggressiveness" of a tumor or cancer
refers to the rate at which the tumor is growing. Thus, a tumor is
more aggressive than another tumor or cancer if it is proliferating
at a higher rate. Other determinants can be used to measure the
level of aggressiveness of a tumor or cancer, for example, based on
the appearance of tumor or cancer cells under a microscope to
determine the extent to which tumors are differentiated. A
well-differentiated tumor tends to be more aggressive than a
poorly-differentiated tumor or cancer.
[0150] The term "selective inhibition" or "selectively inhibit" as
applied to a biologically active agent refers to the agent's
ability to selectively reduce the target signaling activity as
compared to off-target signaling activity, via direct or indirect
interaction with the target. For example, a compound that
selectively inhibits one isoform of PI3K over another isoform of
PI3K has an activity of at least greater than about 1.times.
against a first isoform relative to the compound's activity against
the second isoform (e.g., at least about 2.times., 3.times.,
5.times., 10.times., 20.times., 50.times., 100.times., 200.times.,
500.times., or 1000.times.). In certain embodiments, these terms
refer to (1) a compound described herein that selectively inhibits
the gamma isoform over the alpha, beta, or delta isoform; or (2) a
compound described herein that selectively inhibits the delta
isoform over the alpha or beta isoform. By way of non-limiting
example, the ratio of selectivity can be greater than a factor of
about 1, greater than a factor of about 2, greater than a factor of
about 3, greater than a factor of about 5, greater than a factor of
about 10, greater than a factor of about 50, greater than a factor
of about 100, greater than a factor of about 200, greater than a
factor of about 400, greater than a factor of about 600, greater
than a factor of about 800, greater than a factor of about 1000,
greater than a factor of about 1500, greater than a factor of about
2000, greater than a factor of about 5000, greater than a factor of
about 10,000, or greater than a factor of about 20,000, where
selectivity can be measured by IC.sub.50. In certain embodiments,
the IC.sub.50 can be measured by in vitro or in vivo assays. In
certain embodiments, the PI3K gamma isoform IC.sub.50 activity of a
compound provided herein can be less than about 1000 nM, less than
about 500 nM, less than about 400 nM, less than about 300 nM, less
than about 200 nM, less than about 100 nM, less than about 75 nM,
less than about 50 nM, less than about 25 nM, less than about 20
nM, less than about 15 nM, less than about 10 nM, less than about 5
nM, or less than about 1 nM. In certain embodiments, the PI3K delta
isoform IC.sub.50 activity of a compound provided herein can be
less than about 1000 nM, less than about 500 nM, less than about
400 nM, less than about 300 nM, less than about 200 nM, less than
about 100 nM, less than about 75 nM, less than about 50 nM, less
than about 25 nM, less than about 20 nM, less than about 15 nM,
less than about 10 nM, less than about 5 nM, or less than about 1
nM.
[0151] "Subject" or "patient" to which administration is
contemplated includes, but is not limited to, humans (e.g., a male
or female of any age group, e.g., a pediatric subject (e.g.,
infant, child, adolescent) or adult subject (e.g., young adult,
middle-aged adult or senior adult)) and/or other primates (e.g.,
cynomolgus monkeys, rhesus monkeys); mammals, including
commercially relevant mammals such as cattle, pigs, horses, sheep,
goats, cats, and/or dogs; and/or birds, including commercially
relevant birds such as chickens, ducks, geese, quail, and/or
turkeys.
[0152] The term "in vivo" refers to an event that takes place in a
subject's body.
[0153] The term "in vitro" refers to an event that takes places
outside of a subject's body. For example, an in vitro assay
encompasses any assay conducted outside of a subject. In vitro
assays encompass cell-based assays in which cells, alive or dead,
are employed. In vitro assays also encompass a cell-free assay in
which no intact cells are employed.
[0154] Combination therapy, or "in combination with" refer to the
use of more than one compound or agent to treat a particular
disorder or condition. For example, a PI3K inhibitor (e.g.,
Compound 1) may be administered in combination with at least one
additional therapeutic agent (e.g. a Bcl-2 inhibitor). By "in
combination with," it is not intended to imply that the PI3K
inhibitor and additional therapeutic agent (e.g., the BLC-2
inhibitor) must be administered at the same time and/or formulated
for delivery together, although these methods of delivery are
within the scope of this disclosure. The PI3K inhibitor, e.g.,
Compound 1, can be administered concurrently with, 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, 12
weeks, or 16 weeks before), 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, 12 weeks, or 16 weeks
after), one or more other additional agents. In certain
embodiments, each therapeutic agent will be administered at a dose
and/or on a time schedule determined for that particular agent. The
other therapeutic agent (e.g., the Bcl-2 inhibitor) can be
administered with the PI3K inhibitor (e.g., Compound 1) herein in a
single composition or separately in a different composition. Higher
combinations, e.g., triple therapy, are also contemplated
herein.
[0155] The terms "co-administration of" and "co-administering" and
their grammatical equivalents, as used herein, encompass
administration of two or more agents to subject so that both agents
and/or their metabolites are present in the subject at the same or
substantially the same time. In one embodiment, co-administration
of a PI3K inhibitor with an additional anti-cancer agent (both
components referred to hereinafter as the "two active agents")
refer to any administration of the two active agents, either
separately or together, where the two active agents are
administered as part of an appropriate dose regimen designed to
obtain the benefit of the combination therapy. Thus, the two active
agents can be administered either as part of the same
pharmaceutical composition or in separate pharmaceutical
compositions. The additional agent can be administered prior to, at
the same time as, or subsequent to administration of the PI3K
inhibitor, or in some combination thereof. Where the PI3K inhibitor
is administered to the patient at repeated intervals, e.g., during
a standard course of treatment, the additional agent can be
administered prior to, at the same time as, or subsequent to, each
administration of the PI3K inhibitor, or some combination thereof,
or at different intervals in relation to the PI3K inhibitor
treatment, or in a single dose prior to, at any time during, or
subsequent to the course of treatment with the PI3K inhibitor. In
certain embodiments, a first agent can be 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), essentially 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 a second
therapeutic agent.
[0156] As used herein, a "monotherapy" refers to the use of an
agent individually (e.g., as a single compound or agent), e.g.,
without a second active agent to treat the same indication, e.g.,
cancer. For example, in this context, the term monotherapy includes
the use of either the PI3K inhibitor or the second agent
individually to treat the cancer.
[0157] The term "synergy" or "synergistic" encompasses a more than
additive effect of a combination of two or more agents compared to
their individual effects. In certain embodiments, synergy or
synergistic effect refers to an advantageous effect of using two or
more agents in combination, e.g., in a pharmaceutical composition,
or in a method of treatment. In certain embodiments, one or more
advantageous effects is achieved by using a PI3K inhibitor in
combination with a second therapeutic agent (e.g., one or Bcl-2
inhibitors) as described herein.
[0158] In some embodiments, the synergistic effect is that a lower
dosage of one or both of the agents is needed to achieve an effect.
For example, the combination can provide a selected effect, e.g., a
therapeutic effect, when at least one of the agents is administered
at a lower dosage than the dose of that agent that would be
required to achieve the same therapeutic effect when the agent is
administered as a monotherapy. In certain embodiments, the
combination of a PI3K inhibitor (e.g., Compound 1) and a second
agent as described herein (e.g., a Bcl-2 inhibitor as described
herein) allows the PI3K inhibitor to be administered at a lower
dosage than would be required to achieve the same therapeutic
effect if the PI3K inhibitor were administered as a monotherapy. In
certain embodiments, the combination of a PI3K inhibitor (e.g.,
Compound 1) and the Bcl-2 inhibitor allows the Bcl-2 inhibitor to
be administered at a lower dosage than would be required to achieve
the same therapeutic effect if the Bcl-2 inhibitor were
administered as a monotherapy.
[0159] In some embodiments, the synergistic effect is a reduction,
prevention, delay, or decrease in the occurrence or the likelihood
of occurrence of one or more side effects, toxicity, resistance,
that would otherwise be associated with administration of at least
one of the agents.
[0160] In some embodiments, the synergistic effect is a reduction
in resistance (e.g., a decrease in a measure of resistance or a
decreased likelihood of developing resistance), or a delay in the
development of resistance, to at least one of the agents.
[0161] In some embodiments, the synergistic effect is a reduction
in MRD. In certain embodiments, the combination of a PI3K inhibitor
(e.g. a PI3K inhibitor described herein) and a second agent (e.g.,
a Bcl-2 inhibitor) is effective to reduce the MRD in the subject,
e.g., below a level previously measured in the subject (e.g., the
level measured before the combination was administered). In certain
embodiments, the combination of a PI3K inhibitor and a second agent
(e.g., a Bcl-2 inhibitor) is effective to reduce the MRD in the
subject below the level observed during or after treatment with a
monotherapy, e.g., a monotherapy comprising either the PI3K
inhibitor or the second agent (e.g., the Bcl-2 inhibitor). In
certain embodiments, the MRD is decreased below the level observed
during treatment with a monotherapy comprising the PI3K inhibitor.
In certain embodiments, the MRD is decreased below the level
observed during treatment with a monotherapy comprising the second
agent (e.g., the Bcl-2 inhibitor). In certain embodiments, the
combination is effective to reduce the MRD below a preselected
cutoff value (e.g., 1 malignant cell in 100 normal cells, 1
malignant cell in 1000 normal cells, or 1 malignant cell in 10,000
normal cells). In certain embodiments, the preselected cutoff value
is 1 malignant cell in 1000 or 10,000 normal cells.
[0162] In some embodiments, a synergistic effect refers to the
combination of a PI3K inhibitor (e.g., Compound 1, or a
pharmaceutically acceptable form thereof), and a second therapeutic
agent (e.g., one or more the Bcl-2 inhibitor(s), or a
pharmaceutically acceptable forms thereof, as described herein),
results in a therapeutic effect greater than the additive effect of
the PI3K inhibitor and the second agent.
[0163] In some embodiments, a synergistic effect means that
combination index value is less than a selected value, e.g., for a
given effect, e.g., at a selected percentage (e.g., 50%) inhibition
or growth inhibition, e.g., as described herein in the Examples. In
certain embodiments, the selected value is 1. In certain
embodiments, the selected value is 0.7. In certain embodiments, the
selected value is 0.5.
[0164] In some embodiments, a synergistic effect means that the
synergy score is 1 or more. In certain embodiments, the synergy
score is greater than 1. In certain embodiments, the synergy score
is 3 or more. In certain embodiments, the synergy score is 10 or
more.
[0165] Combination index (CI) is a score of the potency shifting.
Chou et al., Adv Enzyme Regul 1984; 22: 27-55 and in U.S. Patent
Publication No. 2013/0295102, the contents of which are
incorporated herein by reference. A CI value of greater than 1
indicates antagonistic effect; a CI value of 1.0 is indicative of
an additive effect; and a CI value of less than 1 is indicative of
a synergistic effect resulting from the combination. The CI value
can be determined at various percentages of inhibition or growth
inhibition.
[0166] The CI can be thought of as an estimate of the fraction of
the original (monotherapy) doses of each of two drugs would be
needed in combination relative to the single agent doses required
to achieve a chosen effect level. For example, when the combination
index has a value of 0.1, only about one tenth of the total
fractional amounts of the individual agents (expressed as a
fraction of the amount of that agent when administered as a
monotherapy to achieve a chosen effect) are needed for the
combination to reach the same chosen effect level. For example, if
a dose of 100 mg/kg of drug A individually or a dose of 200 mg/kg
of drug B individually is needed to achieve the chosen effect, and
the combination index is 0.1, then approximately 5 mg/kg of drug A
and 10 mg/kg of drug B would achieve the chosen effect (one
twentieth of the original doses of each of the single agents adds
up to a total of one tenth). The doses of the single agents need
not be reduced by the same fractional value so long as the sum of
their fractional values adds up to the combination index; thus, in
this example, a dose of approximately 8 mg/kg of drug A and 4 mg/kg
of drug B would also achieve the chosen effect (this is 0.08 times
the original dose of drug A and 0.02 times the original dose of
drug B; the sum of the fractional amounts (0.08+0.02) is equal to
the combination index of 0.1.)
[0167] According to one embodiment, synergy score is a measure of
the combination effects in excess of Loewe additivity. In one
example, synergy score is a scalar measure to characterize the
strength of synergistic interaction. The Synergy score can be
calculated as:
Synergy Score=log f.sub.X log
f.sub.Y.SIGMA.max(0,I.sub.data)(I.sub.data-I.sub.Loewe)
In this example, the fractional inhibition for each component agent
and combination point in the matrix is calculated relative to the
median of all vehicle-treated control wells. The example Synergy
Score equation integrates the experimentally-observed activity
volume at each point in the matrix in excess of a model surface
numerically derived from the activity of the component agents using
the Loewe model for additivity. Additional terms in the Synergy
Score equation (above) are used to normalize for various dilution
factors used for individual agents and to allow for comparison of
synergy scores across an entire experiment. The inclusion of
positive inhibition gating or an I.sub.data multiplier removes
noise near the zero effect level, and biases results for
synergistic interactions at that occur at high activity levels.
According to other embodiments, a synergy score can be calculated
based on a curve fitting approach where the curvature of the
synergy score is extrapolated by introducing a median value and
origin value (e.g., a dose zero value).
[0168] The synergy score measure can be used for the self-cross
analysis. Synergy scores of self-crosses are expected to be
additive by definition and, therefore, maintain a synergy score of
zero. However, while some self-cross synergy scores are near zero,
many are greater suggesting that experimental noise or non-optimal
curve fitting of the single agent dose responses are contributing
to the slight perturbations in the score. This strategy is cell
line-centric, focusing on self-cross behavior in each cell line
versus a global review of cell line panel activity. Combinations
where the synergy score is greater than the mean self-cross plus
two standard deviations or three standard deviations can be
considered candidate synergies at 95% and 99% confidence levels,
respectively. Additivity should maintain a synergy score of zero,
and synergy score of two or three standard deviations indicate
synergism at statistically significant levels of 95% and 99%.
[0169] Loewe Volume (Loewe Vol) is used to assess the overall
magnitude of the combination interaction in excess of the Loewe
additivity model. Loewe Volume is particularly useful when
distinguishing synergistic increases in a phenotypic activity
(positive Loewe Volume) versus synergistic antagonisms (negative
Loewe Volume). When antagonisms are observed, the Loewe Volume
should be assessed to examine if there is any correlation between
antagonism and a particular drug target-activity or cellular
genotype. This model defines additivity as a non-synergistic
combination interaction where the combination dose matrix surface
should be indistinguishable from either drug crossed with itself.
The calculation for Loewe additivity is:
I.sub.Loewe that satisfies(X/X.sub.I)+(Y/Y.sub.I)=1
where X.sub.I and Y.sub.I are the single agent effective
concentrations for the observed combination effect I. For example,
if 50% inhibition is achieved separately by 1 .mu.M of drug A or 1
.mu.M of drug B, a combination of 0.5 .mu.M of A and 0.5 .mu.M of B
should also inhibit by 50%.
[0170] As used herein, a daily dosage can be achieved by a single
administration of the targeted dosage amount or multiple
administrations of smaller dosage amount(s). For example, a 150 mg
daily dosage can be achieved by a single administration of 150 mg
of the therapeutic agent per day, two administrations of 75 mg of
the therapeutic agent per day, or three administrations of 50 mg of
the therapeutic agent per day, or the like.
[0171] The term "anti-cancer effect" refers to the effect a
therapeutic agent has on cancer, e.g., a decrease in growth,
viability, or both of a cancer cell. The IC.sub.50 of cancer cells
can be used as a measure the anti-cancer effect.
[0172] IC.sub.50 refers to a measure of the effectiveness of a
therapeutic agent in inhibiting cancer cells by 50%.
[0173] The term "tumor" refers to any neoplastic cell growth and
proliferation, whether malignant or benign, and any pre-cancerous
and cancerous cells and tissues. As used herein, the term
"neoplastic" refers to any form of dysregulated or unregulated cell
growth, whether malignant or benign, resulting in abnormal tissue
growth. Thus, "neoplastic cells" include malignant and benign cells
having dysregulated or unregulated cell growth.
[0174] The term "cancer" includes, but is not limited to, solid
tumors and blood born tumors. The term "cancer" refers to disease
of skin tissues, organs, blood, and vessels, including, but not
limited to, cancers of the bladder, bone or blood, brain, breast,
cervix, chest, colon, endometrium, esophagus, eye, head, kidney,
liver, lymph nodes, lung, mouth, neck, ovaries, pancreas, prostate,
rectum, stomach, testis, throat, and uterus.
[0175] Hematopoietic origin refers to involving cells generated
during hematopoiesis, a process by which cellular elements of
blood, such as lymphocytes, leukocytes, platelets, erythrocytes and
natural killer cells are generated. Cancers of hematopoietic origin
includes lymphoma and leukemia.
[0176] Resistant or refractive refers to when a cancer that has a
reduced responsiveness to a treatment, e.g., up to the point where
the cancer does not respond to treatment. The cancer can be
resistant at the beginning of treatment, or it may become resistant
during treatment. The cancer subject may have one or more mutations
that cause it to become resistant to the treatment, or the subject
may have developed such mutations during treatment. The term
"refractory" can refer to a cancer for which treatment (e.g.
chemotherapy drugs, biological agents, and/or radiation therapy)
has proven to be ineffective. A refractory cancer tumor may shrink,
but not to the point where the treatment is determined to be
effective. Typically however, the tumor stays the same size as it
was before treatment (stable disease), or it grows (progressive
disease).
[0177] "Acquire" or "acquiring" as the terms are used herein, refer
to obtaining possession of, determining, or evaluating, a value or
information (e.g., one or more of: the presence, absence, amount or
level) of an alteration or biomarker, by "directly acquiring" or
"indirectly acquiring" the same. "Directly acquiring" means
performing a process (e.g., performing a test) to obtain the value
or information of the alteration or biomarker. "Indirectly
acquiring" refers to receiving the value or information of the
alteration or biomarker from another party or source (e.g., a
diagnostic provider, a third party clinician or health
professional).
[0178] "Alteration" of a gene or gene product (e.g., a biomarker
gene or gene product) or an "altered gene" or "altered gene
product" as used herein, refers to the presence of a mutation
(e.g., one or more mutations) within a gene or gene product, which
affects the structure, amount or activity of the gene or gene
product, as compared to a reference gene or gene product, e.g., a
normal or wild-type gene or gene product, or a responder gene or
gene product (e.g., a gene or gene product in a responder subject
(e.g., a subject in complete or partial cancer remission)). The
alteration can be in amount, structure, and/or activity in a cancer
tissue or cancer cell, as compared to its amount, structure, and/or
activity, in a reference tissue or cell (e.g., a normal or healthy
tissue or cell, or a responder tissue or cell (e.g., a tissue or
cell from a subject in complete or partial cancer remission)). The
alteration can be associated with, or be indicative of, a disease
state, such as cancer (e.g., a hematologic malignancy as described
herein, e.g., CLL). For example, an alteration which is associated
with cancer, or is predictive of responsiveness or
non-responsiveness to an anti-cancer therapeutic (e.g., a PI3K
inhibitor disclosed herein), can have an altered nucleotide
sequence (e.g., a mutation), amino acid sequence, chromosomal
translocation, intra-chromosomal inversion, copy number, expression
level, protein level, protein activity, or methylation status, in a
cancer tissue or cancer cell, as compared to a reference tissue or
cell. Exemplary mutations include, but are not limited to, point
mutations (e.g., silent, missense, or nonsense), deletions,
insertions, inversions, linking mutations, duplications, copy
number changes, translocations, inter- and intra-chromosomal
rearrangements. Mutations can be present in the coding or
non-coding region of the gene (e.g., one or more exons, the 5'-
and/or 3'-UTR).
[0179] In certain embodiments, the alteration(s) are associated (or
not associated) with a phenotype, e.g., a cancerous phenotype
(e.g., one or more of cancer risk; cancer progression;
responsiveness to a cancer treatment (e.g., complete or partial
remission); or decreased responsiveness or non-responsiveness to a
cancer treatment (e.g., progressive or stable disease, or
resistance, e.g., acquired resistance) to a cancer treatment). In
one embodiment, the alteration is associated with, or is, a
prognosis-positive predictor or a prognosis-negative predictor
(also referred to herein as a "prognosis-positive alteration" or a
"prognosis-negative alteration"). In another embodiment, the
alteration is associated with, or is, a progression-positive
predictor or a progression-negative predictor (also referred to
herein as a "progression-positive alteration" or a
"progression-negative alteration").
[0180] As used herein, the term `prognosis-positive predictor`
refers to any alteration that indicates increased responsiveness
(e.g., increased sensitivity) to a PI3K inhibitor. The
prognosis-positive predictor can be evaluated relative to a
reference value, e.g., a normal or wild-type gene or gene product,
or a responder gene or gene product (e.g., a gene or gene product
in a responder subject (e.g., a subject in complete or partial
cancer remission)). Subjects in complete or partial cancer
remission (e.g., CR or PR subjects as described herein) can have
one or more prognosis-positive alterations.
[0181] The term `prognosis-negative predictor` refers to any
alteration that indicates decreased responsiveness (e.g.,
sensitivity) to a PI3K inhibitor. The prognosis-negative predictor
can be evaluated relative to a reference value, e.g., a reference
value disclosed herein. Subjects with progressive disease or stable
disease (e.g., PD or SD subjects as described herein) can have one
or more prognosis-negative alterations. This term can include a
subject who has resistance (e.g., has developed or acquired
resistance) to a PI3K inhibitor.
[0182] The term `progression-positive predictor` refers to any
alteration that indicates increased progression or increased
likelihood of cancer progression. The progression-positive
predictor can be evaluated relative to a reference value, e.g., a
reference value disclosed herein. Subjects with progressive disease
or stable disease (e.g., PD or SD subjects as described herein) can
have one or more progression-positive alterations. This term can
include a subject who has resistance (e.g., has developed or
acquired resistance) to a PI3K inhibitor.
[0183] The term `progression-negative predictor` refers to any
alteration that indicates decreased progression or decreased
likelihood of cancer progression. The progression-negative
predictor can be evaluated relative to a reference value, e.g., a
reference value disclosed herein. Subjects in complete or partial
cancer remission (e.g., CR or PR subjects as described herein) can
have one or more progression-negative alterations.
[0184] A "biomarker" or "marker" is a substance, e.g., a gene or
gene product (e.g., mRNA or protein) which can be altered (e.g.,
having an alteration described herein), wherein said alteration is
associated with, or is indicative of, a disease state, e.g., a
cancer (e.g., a hematological malignancy described herein, e.g.,
CLL). The alteration can be in amount, structure, and/or activity
of the substance (e.g., gene or gene product) in a cancer tissue or
cancer cell, as compared to its amount, structure, and/or activity,
in a reference sample, e.g., a normal or wild-type gene or gene
product, or a responder gene or gene product (e.g., a gene or gene
product in a responder subject (e.g., a subject in complete or
partial cancer remission). For example, a biomarker described
herein which is associated with cancer or predictive of
responsiveness to anti-cancer therapeutics can have an altered
nucleotide sequence, amino acid sequence, chromosomal
translocation, intra-chromosomal inversion, copy number, expression
level, protein level, protein activity, or methylation status, in a
cancer tissue or cancer cell as compared to a normal, healthy
tissue or cell. Furthermore, a "biomarker" includes a molecule
whose structure is altered, e.g., mutated (contains an mutation),
e.g., differs from the wild type sequence at the nucleotide or
amino acid level, e.g., by substitution, deletion, or insertion,
when present in a tissue or cell associated with a disease state,
such as cancer. In some embodiments, a biomarker can be evaluated
individually, or in combinations with one or more other
biomarkers.
[0185] As used herein, the term `prognosis-positive biomarker`
refers to any biomarker that indicates increased responsiveness
(e.g., increased sensitivity) to a PI3K inhibitor. The
prognosis-positive biomarker can be evaluated relative to a
reference value, e.g., a normal or wild-type gene or gene product,
or a responder gene or gene product (e.g., a gene or gene product
in a responder subject (e.g., a subject in complete or partial
cancer remission)). Subjects in complete or partial cancer
remission (e.g., CR or PR subjects as described herein) can have
one or more prognosis-positive biomarkers.
[0186] The term `prognosis-negative biomarker` refers to any
biomarker that indicates decreased responsiveness (e.g.,
sensitivity) to a PI3K inhibitor. The prognosis-negative biomarker
can be evaluated relative to a reference value, e.g., a reference
value disclosed herein. Subjects with progressive disease or stable
disease (e.g., PD or SD subjects as described herein) can have one
or more prognosis-negative biomarkers. This term can include a
subject who has resistance (e.g., has developed or acquired
resistance) to a PI3K inhibitor.
[0187] The term `progression-positive biomarker` refers to any
biomarker that indicates increased progression or increased
likelihood of cancer progression. The progression-positive
biomarker can be evaluated relative to a reference value, e.g., a
reference value disclosed herein. Subjects with progressive disease
or stable disease (e.g., PD or SD subjects as described herein) can
have one or more progression-positive biomarker. This term can
include a subject who has resistance (e.g., has developed or
acquired resistance) to a PI3K inhibitor.
[0188] The term `progression-negative biomarker` refers to any
biomarker that indicates decreased progression or decreased
likelihood of cancer progression. The progression-negative
biomarker can be evaluated relative to a reference value, e.g., a
reference value disclosed herein. Subjects in complete or partial
cancer remission (e.g., CR or PR subjects as described herein) can
have one or more progression-negative biomarkers.
[0189] One skilled in the art can recognize that a prognostic
biomarker may be used as a diagnostic biomarker or a predictive
biomarker, and terms such as `prognosis-positive`,
`prognosis-negative`, `progression-positive` and
progression-negative' and the like may refer to biomarkers used in
methods involving prediction or diagnosis.
[0190] "Copy number loss" as used herein refers to the loss of one
or more copies of a DNA sequence from a genome. In some
embodiments, the DNA sequence comprises a gene. In some
embodiments, the DNA sequence comprises a portion of a gene, e.g.,
such that loss of the portion reduces or abrogates the gene
function. In some embodiments, copy number loss is a result of a
deletion, chromosome loss, or chromosome breakage event.
[0191] "Responsiveness," to "respond" to treatment, and other forms
of this term, as used herein, refer to the reaction of a subject to
treatment with a therapeutic, e.g., a PI3K inhibitor, alone or in
combination, e.g., monotherapy or combination therapy. In one
embodiment, a response to a PI3K inhibitor is determined.
Responsiveness to a therapy, e.g., treatment with a PI3K inhibitor
alone or in combination, can be evaluated by using any of the
alterations/biomarkers disclosed herein and/or comparing a
subject's response to the therapy using one or more clinical
criteria, such as IWCLL 2008 (for CLL) described in, e.g., Hallek,
M. et al. (2008) Blood 111 (12): 5446-5456; RECIST criteria for
solid tumors (Response Evaluation Criteria In Solid Tumors), and
the like. Additional classifications of responsiveness are provided
in Brown, J. R. (2014) Blood, 123(22):3390-3397 and Chesson, B. D.
et al. Journal of Clinical Oncology, 30(23):2820-2822.
[0192] These criteria provide a set of published rules that define
when cancer patients improve ("respond"), stay the same ("stable")
or worsen ("progression") during treatments.
[0193] In one embodiment, a subject having CLL can be determined to
be in complete remission (CR) or partial remission (PR). For
example, according to IWCLL 2008, a subject is considered to be in
CR if at least all of the following criteria as assessed after
completion of therapy are met: (i) Peripheral blood lymphocytes
(evaluated by blood and different count) below 4.times.10.sup.9/L
(4000 .mu.L); (ii) no hepatomegaly or splenomegaly by physical
examination; (iii) absence of constitutional symptoms; and (iv)
blood counts (e.g., neutrophils, platelets, hemoglobin) above the
values set forth in Hallek, M. et al. supra at page 5451). Partial
remission (PR) for CLL is defined according to IWCLL 2008 as
including one of: (i) a decrease in number of blood lymphocytes by
50% or more from the value before therapy; (ii) a reduction in
lymphadenopathy, as detected by CT scan or palpation; or (iii) a
reduction in pretreatment enlargement of spleen or liver by 50% or
more, as detected by CT scan or palpation; and blood counts (e.g.,
neutrophils, platelets, hemoglobin) according to the values set
forth in Hallek, M. et al. supra at page 5451).
[0194] In other embodiments, a subject having CLL is determined to
have progressive disease (PD) or stable disease (SD). For example,
according to IWCLL 2008, a subject is considered to be in PD during
therapy or after therapy if at least one of the following criteria
is met: (i) progression on lymphadenopathy; (ii) an increase in
pretreatment enlargement of spleen or liver by 50% or more, or de
novo appearance of hepatomegaly or splenomegaly; (iii) an increase
in the number of blood lymphocytes by 50% or more with at least
5000 B lymphocytes per microliter; (iv) transformation to a more
aggressive histology (e.g., Richter syndrome); or (v) occurrence of
cytopenia (neutropenia, anemia or thrombocytopenia) attributable to
CLL, as described in Hallek, M. et al. supra at page 5452. Stable
disease (SD) for CLL is defined according to IWCLL 2008 as a
patient who has not achieved CR or a PR, and who has not exhibited
progressive disease, see Hallek, M. et al. supra at page 5452.
[0195] In one embodiment, a subject with CLL responds to treatment
with a PI3K inhibitor if at least one of the criteria for disease
progression according to IWCLL is retarded or reduced, e.g., by
about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more. In
another example, a subject responds to treatment with a PI3K
inhibitor, if the subject experiences a life expectancy extension,
e.g., extended by about 5%, 10%, 20%, 30%, 40%, 50% or more beyond
the life expectancy predicted if no treatment is administered. In
another example, a subject responds to treatment with a PI3K
inhibitor, if the subject has one or more of: an increased
progression-free survival, overall survival or increased time to
progression (TTP), e.g., as described in Hallek, M. et al. supra at
page 5452.
[0196] In another embodiment in solid tumors, a subject responds to
treatment with a PI3K inhibitor if growth of a tumor in the subject
is retarded about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or
more. In another example, a subject responds to treatment with a
PI3K inhibitor, if a tumor in the subject shrinks by about 5%, 10%,
20%, 30%, 40%, 50% or more as determined by any appropriate
measure, e.g., by mass or volume. In another example, a subject
responds to treatment with a PI3K inhibitor, if the subject
experiences a life expectancy extended by about 5%, 10%, 20%, 30%,
40%, 50% or more beyond the life expectancy predicted if no
treatment is administered. In another example, a subject responds
to treatment with a PI3K inhibitor, if the subject has an increased
disease-free survival, overall survival or increased time to
progression. Several methods can be used to determine if a patient
responds to a treatment including the RECIST criteria, as set forth
above.
Chemical Definitions
[0197] As used herein, a "pharmaceutically acceptable form" of a
disclosed compound includes, but is not limited to,
pharmaceutically acceptable salts, hydrates, solvates, isomers,
prodrugs, and isotopically labeled derivatives of disclosed
compounds. In one embodiment, a "pharmaceutically acceptable form"
includes, but is not limited to, pharmaceutically acceptable salts,
isomers, prodrugs and isotopically labeled derivatives of disclosed
compounds.
[0198] In certain embodiments, the pharmaceutically acceptable form
is a pharmaceutically acceptable salt. As used herein, the term
"pharmaceutically acceptable salt" refers to those salts which are,
within the scope of sound medical judgment, suitable for use in
contact with the tissues of subjects without undue toxicity,
irritation, allergic response and the like, and are commensurate
with a reasonable benefit/risk ratio. Pharmaceutically acceptable
salts are well known in the art. For example, Berge et al.
describes pharmaceutically acceptable salts in detail in J.
Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable
salts of the compounds provided herein include those derived from
suitable inorganic and organic acids and bases. Examples of
pharmaceutically acceptable, nontoxic acid addition salts are salts
of an amino group formed with inorganic acids such as hydrochloric
acid, hydrobromic acid, phosphoric acid, sulfuric acid and
perchloric acid or with organic acids such as acetic acid, oxalic
acid, maleic acid, tartaric acid, citric acid, succinic acid or
malonic acid or by using other methods used in the art such as ion
exchange. Other pharmaceutically acceptable salts include adipate,
alginate, ascorbate, aspartate, benzenesulfonate, besylate,
benzoate, bisulfate, borate, butyrate, camphorate,
camphorsulfonate, citrate, cyclopentanepropionate, digluconate,
dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate,
glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate,
hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate,
laurate, lauryl sulfate, malate, maleate, malonate,
methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,
oleate, oxalate, palmitate, pamoate, pectinate, persulfate,
3-phenylpropionate, phosphate, picrate, pivalate, propionate,
stearate, succinate, sulfate, tartrate, thiocyanate,
p-toluenesulfonate, undecanoate, valerate salts, and the like. In
some embodiments, organic acids from which salts may be derived
include, for example, acetic acid, propionic acid, glycolic acid,
pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic
acid, fumaric acid, tartaric acid, citric acid, benzoic acid,
cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic
acid, p-toluenesulfonic acid, salicylic acid, and the like.
[0199] Pharmaceutically acceptable salts derived from appropriate
bases include alkali metal, alkaline earth metal, ammonium and
N.sup.+(C.sub.1-4alkyl).sub.4 salts. Representative alkali or
alkaline earth metal salts include sodium, lithium, potassium,
calcium, magnesium, iron, zinc, copper, manganese, aluminum, and
the like. Further pharmaceutically acceptable salts include, when
appropriate, nontoxic ammonium, quaternary ammonium, and amine
cations formed using counterions such as halide, hydroxide,
carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate,
and aryl sulfonate. Organic bases from which salts may be derived
include, for example, primary, secondary, and tertiary amines,
substituted amines including naturally occurring substituted
amines, cyclic amines, basic ion exchange resins, and the like,
such as isopropylamine, trimethylamine, diethylamine,
triethylamine, tripropylamine, and ethanolamine. In some
embodiments, the pharmaceutically acceptable base addition salt is
chosen from ammonium, potassium, sodium, calcium, and magnesium
salts.
[0200] In certain embodiments, the pharmaceutically acceptable form
is a solvate (e.g., a hydrate). As used herein, the term "solvate"
refers to compounds that further include a stoichiometric or
non-stoichiometric amount of solvent bound by non-covalent
intermolecular forces. The solvate may be of a disclosed compound
or a pharmaceutically acceptable salt thereof. Where the solvent is
water, the solvate is a "hydrate". Pharmaceutically acceptable
solvates and hydrates are complexes that, for example, can include
1 to about 100, or 1 to about 10, or one to about 2, about 3 or
about 4, solvent or water molecules. It will be understood that the
term "compound" as used herein encompasses the compound and
solvates of the compound, as well as mixtures thereof.
[0201] In certain embodiments, the pharmaceutically acceptable form
is a prodrug. As used herein, the term "prodrug" refers to
compounds that are transformed in vivo to yield a disclosed
compound or a pharmaceutically acceptable form of the compound. A
prodrug may be inactive when administered to a subject, but is
converted in vivo to an active compound, for example, by hydrolysis
(e.g., hydrolysis in blood). In certain cases, a prodrug has
improved physical and/or delivery properties over the parent
compound. Prodrugs are typically designed to enhance
pharmaceutically and/or pharmacokinetically based properties
associated with the parent compound. The prodrug compound often
offers advantages of solubility, tissue compatibility or delayed
release in a mammalian organism (see, e.g., Bundgard, H., Design of
Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam). A discussion
of prodrugs is provided in Higuchi, T., et al., "Pro-drugs as Novel
Delivery Systems," A.C.S. Symposium Series, Vol. 14, Chp 1, pp 1-12
and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche,
American Pharmaceutical Association and Pergamon Press, 1987, both
of which are incorporated in full by reference herein. Exemplary
advantages of a prodrug can include, but are not limited to, its
physical properties, such as enhanced water solubility for
parenteral administration at physiological pH compared to the
parent compound, or it enhances absorption from the digestive
tract, or it can enhance drug stability for long-term storage.
[0202] The term "prodrug" is also meant to include any covalently
bonded carriers, which release the active compound in vivo when
such prodrug is administered to a subject. Prodrugs of an active
compound, as described herein, may be prepared by modifying
functional groups present in the active compound in such a way that
the modifications are cleaved, either in routine manipulation or in
vivo, to the parent active compound. Prodrugs include compounds
wherein a hydroxy, amino or mercapto group is bonded to any group
that, when the prodrug of the active compound is administered to a
subject, cleaves to form a free hydroxy, free amino or free
mercapto group, respectively. Examples of prodrugs include, but are
not limited to, acetate, formate and benzoate derivatives of an
alcohol or acetamide, formamide and benzamide derivatives of an
amine functional group in the active compound and the like. Other
examples of prodrugs include compounds that comprise --NO,
--NO.sub.2, --ONO, or --ONO.sub.2 moieties. Prodrugs can typically
be prepared using well-known methods, such as those described in
Burger's Medicinal Chemistry and Drug Discovery, 172-178, 949-982
(Manfred E. Wolff ed., 5th ed., 1995), and Design of Prodrugs (H.
Bundgaard ed., Elsevier, New York, 1985).
[0203] For example, if a disclosed compound or a pharmaceutically
acceptable form of the compound contains a carboxylic acid
functional group, a prodrug can comprise a pharmaceutically
acceptable ester formed by the replacement of the hydrogen atom of
the acid group with a group such as (C.sub.1-C.sub.8)alkyl,
(C.sub.2-C.sub.12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having
from 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having
from 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having from 3 to
6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having from 4 to 7
carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5 to
8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3 to 9
carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4 to 10
carbon atoms, 3-phthalidyl, 4-crotonolactonyl,
gamma-butyrolacton-4-yl,
di-N,N--(C.sub.1-C.sub.2)alkylamino(C.sub.2-C.sub.3)alkyl (such as
3-dimethylaminoethyl), carbamoyl-(C.sub.1-C.sub.2)alkyl,
N,N-di(C.sub.1-C.sub.2)alkylcarbamoyl-(C.sub.1-C.sub.2)alkyl and
piperidino-, pyrrolidino- or morpholino(C.sub.2-C.sub.3)alkyl.
[0204] Similarly, if a disclosed compound or a pharmaceutically
acceptable form of the compound contains an alcohol functional
group, a prodrug may be formed by the replacement of the hydrogen
atom of the alcohol group with a group such as
(C.sub.1-C.sub.6)alkanoyloxymethyl,
1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl,
1-methyl-1-((C.sub.1-C.sub.6)alkanoyloxy)ethyl
(C.sub.1-C.sub.6)alkoxycarbonyloxymethyl,
N--(C.sub.1-C.sub.6)alkoxycarbonylaminomethyl, succinoyl,
(C.sub.1-C.sub.6)alkanoyl, .alpha.-amino(C.sub.1-C.sub.4)alkanoyl,
arylacyl and .alpha.-aminoacyl, or
.alpha.-aminoacyl-.alpha.-aminoacyl, where each .alpha.-aminoacyl
group is independently selected from naturally occurring L-amino
acids, P(O)(OH).sub.2, --P(O)(O(C.sub.1-C.sub.6)alkyl).sub.2, and
glycosyl (the radical resulting from the removal of a hydroxyl
group of the hemiacetal form of a carbohydrate).
[0205] If a disclosed compound or a pharmaceutically acceptable
form of the compound incorporates an amine functional group, a
prodrug may be formed by the replacement of a hydrogen atom in the
amine group with a group such as R-carbonyl, RO-carbonyl,
NRR'-carbonyl where R and R' are each independently
(C.sub.1-C.sub.10)alkyl, (C.sub.3-C.sub.7)cycloalkyl, benzyl, a
natural .alpha.-aminoacyl or natural .alpha.-aminoacyl-natural
.alpha.-aminoacyl, --C(OH)C(O)OY.sup.1 wherein Y.sup.1 is H,
(C.sub.1-C.sub.6)alkyl or benzyl, --C(OY.sup.2)Y.sup.3 wherein
Y.sup.2 is (C.sub.1-C.sub.4) alkyl and Y.sup.3 is
(C.sub.1-C.sub.6)alkyl, carboxy(C.sub.1-C.sub.6)alkyl,
amino(C.sub.1-C.sub.4)alkyl or mono-N- or
di-N,N--(C.sub.1-C.sub.6)alkylaminoalkyl, --C(Y.sup.4)Y.sup.5
wherein Y.sup.4 is H or methyl and Y.sup.5 is mono-N- or
di-N,N--(C.sub.1-C.sub.6)alkylamino, morpholino, piperidin-1-yl or
pyrrolidin-1-yl.
[0206] In certain embodiments, the pharmaceutically acceptable form
is an isomer. "Isomers" are different compounds that have the same
molecular formula. "Stereoisomers" are isomers that differ only in
the way the atoms are arranged in space. As used herein, the term
"isomer" includes any and all geometric isomers and stereoisomers.
For example, "isomers" include geometric double bond cis- and
trans-isomers, also termed E- and Z-isomers; R- and S-enantiomers;
diastereomers, (d)-isomers and (l)-isomers, racemic mixtures
thereof; and other mixtures thereof, as falling within the scope of
this disclosure.
[0207] "Enantiomers" are a pair of stereoisomers that are
non-superimposable mirror images of each other. A 1:1 mixture of a
pair of enantiomers is a "racemic" mixture. The term "(..+-..)" is
used to designate a racemic mixture where appropriate.
"Diastereoisomers" are stereoisomers that have at least two
asymmetric atoms, but which are not mirror-images of each other.
The absolute stereochemistry is specified according to the
Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer
the stereochemistry at each chiral carbon may be specified by
either R or S. Resolved compounds whose absolute configuration is
unknown may be designated (+) or (-) depending on the direction
(dextro- or levorotatory) which they rotate plane polarized light
at the wavelength of the sodium D line. Certain of the compounds
described herein contain one or more asymmetric centers and can
thus give rise to enantiomers, diastereomers, and other
stereoisomeric forms that may be defined, in terms of absolute
stereochemistry, as (R)- or (S)-. The present chemical entities,
pharmaceutical compositions and methods are meant to include all
such possible isomers, including racemic mixtures, optically pure
forms and intermediate mixtures. Optically active (R)- and
(S)-isomers may be prepared using chiral synthons or chiral
reagents, or resolved using conventional techniques. When the
compounds described herein contain olefinic double bonds or other
centers of geometric asymmetry, and unless specified otherwise, it
is intended that the compounds include both E and Z geometric
isomers.
[0208] "Enantiomeric purity" as used herein refers to the relative
amounts, expressed as a percentage, of the presence of a specific
enantiomer relative to the other enantiomer. For example, if a
compound, which can potentially have an (R)- or an (S)-isomeric
configuration, is present as a racemic mixture, the enantiomeric
purity is about 50% with respect to either the (R)- or (S)-isomer.
If that compound has one isomeric form predominant over the other,
for example, 80% (S)- and 20% (R)-, the enantiomeric purity of the
compound with respect to the (S)-isomeric form is 80%. The
enantiomeric purity of a compound may be determined in a number of
ways known in the art, including but not limited to chromatography
using a chiral support, polarimetric measurement of the rotation of
polarized light, nuclear magnetic resonance spectroscopy using
chiral shift reagents which include but are not limited to
lanthanide containing chiral complexes or the Pirkle alcohol, or
derivatization of a compounds using a chiral compound such as
Mosher's acid followed by chromatography or nuclear magnetic
resonance spectroscopy.
[0209] In certain embodiments, the pharmaceutically acceptable form
is a tautomer. As used herein, the term "tautomer" is a type of
isomer that includes two or more interconvertable compounds
resulting from at least one formal migration of a hydrogen atom and
at least one change in valency (e.g., a single bond to a double
bond, a triple bond to a double bond, or a triple bond to a single
bond, or vice versa). "Tautomerization" includes prototropic or
proton-shift tautomerization, which is considered a subset of
acid-base chemistry. "Prototropic tautomerization" or "proton-shift
tautomerization" involves the migration of a proton accompanied by
changes in bond order. The exact ratio of the tautomers depends on
several factors, including temperature, solvent, and pH. Where
tautomerization is possible (e.g., in solution), a chemical
equilibrium of tautomers may be reached. Tautomerizations (i.e.,
the reaction providing a tautomeric pair) may be catalyzed by acid
or base, or can occur without the action or presence of an external
agent. Exemplary tautomerizations include, but are not limited to,
keto-enol; amide-imide; lactam-lactim; enamine-imine; and
enamine-(a different) enamine tautomerizations. A specific example
of keto-enol tautomerization is the interconversion of
pentane-2,4-dione and 4-hydroxypent-3-en-2-one tautomers. Another
example of tautomerization is phenol-keto tautomerization. A
specific example of phenol-keto tautomerization is the
interconversion of pyridin-4-ol and pyridin-4(1H)-one
tautomers.
[0210] Unless otherwise stated, structures depicted herein are also
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structures except for the replacement or enrichment of
a hydrogen by deuterium or tritium at one or more atoms in the
molecule, or the replacement or enrichment of a carbon by .sup.13C
or .sup.14C at one or more atoms in the molecule, are within the
scope of this disclosure. In one embodiment, provided herein are
isotopically labeled compounds having one or more hydrogen atoms
replaced by or enriched by deuterium. In one embodiment, provided
herein are isotopically labeled compounds having one or more
hydrogen atoms replaced by or enriched by tritium. In one
embodiment, provided herein are isotopically labeled compounds
having one or more carbon atoms replaced or enriched by .sup.13C.
In one embodiment, provided herein are isotopically labeled
compounds having one or more carbon atoms replaced or enriched by
.sup.14C.
[0211] The disclosure also embraces isotopically labeled compounds
which are identical to those recited herein, except that one or
more atoms are replaced by an atom having an atomic mass or mass
number different from the atomic mass or mass number usually found
in nature. Examples of isotopes that may be incorporated into
disclosed compounds include isotopes of hydrogen, carbon, nitrogen,
oxygen, phosphorus, sulfur, fluorine, and chlorine, such as, e.g.,
.sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O,
.sup.31P, .sup.32P, .sup.35S, .sup.18F, and .sup.36Cl,
respectively. Certain isotopically-labeled disclosed compounds
(e.g., those labeled with .sup.3H and/or .sup.14C) are useful in
compound and/or substrate tissue distribution assays. Tritiated
(i.e., .sup.3H) and carbon-14 (i.e., .sup.14C) isotopes can allow
for ease of preparation and detectability. Further, substitution
with heavier isotopes such as deuterium (i.e., .sup.2H) can afford
certain therapeutic advantages resulting from greater metabolic
stability (e.g., increased in vivo half-life or reduced dosage
requirements). Isotopically labeled disclosed compounds can
generally be prepared by substituting an isotopically labeled
reagent for a non-isotopically labeled reagent. In some
embodiments, provided herein are compounds that can also contain
unnatural proportions of atomic isotopes at one or more of atoms
that constitute such compounds. All isotopic variations of the
compounds as disclosed herein, whether radioactive or not, are
encompassed within the scope of the present disclosure.
[0212] As used herein, and unless otherwise specified, "polymorph"
may be used herein to describe a crystalline material, e.g., a
crystalline form. In certain embodiments, "polymorph" as used
herein are also meant to include all crystalline and amorphous
forms of a compound or a salt thereof, including, for example,
crystalline forms, polymorphs, pseudopolymorphs, solvates,
hydrates, co-crystals, unsolvated polymorphs (including
anhydrates), conformational polymorphs, tautomeric forms,
disordered crystalline forms, and amorphous forms, as well as
mixtures thereof, unless a particular crystalline or amorphous form
is referred to. Compounds of the present disclosure include
crystalline and amorphous forms of those compounds, including, for
example, crystalline forms, polymorphs, pseudopolymorphs, solvates,
hydrates, co-crystals, unsolvated polymorphs (including
anhydrates), conformational polymorphs, tautomeric forms,
disordered crystalline forms, and amorphous forms of the compounds
or a salt thereof, as well as mixtures thereof.
[0213] It should be noted that if there is a discrepancy between a
depicted structure and a name given that structure, the depicted
structure is to be accorded more weight. In addition, if the
stereochemistry of a structure or a portion of a structure is not
indicated with, for example, bold or dashed lines, the structure or
portion of the structure is to be interpreted as encompassing all
stereoisomers of the structure.
[0214] "Pharmaceutically acceptable carrier" or "pharmaceutically
acceptable excipient" includes any and all solvents, dispersion
media, coatings, antibacterial and antifungal agents, isotonic and
absorption delaying agents and the like. The use of such media and
agents for pharmaceutically active substances is well known in the
art. Except insofar as any conventional media or agent is
incompatible with the active ingredient, its use in the therapeutic
compositions as disclosed herein is contemplated. Supplementary
active ingredients can also be incorporated into the pharmaceutical
compositions.
[0215] As used herein, a "phosphoinositide 3-kinase (PI3K)
inhibitor" or "PI3K inhibitor" refers to an inhibitor of any PI3K.
PI3Ks are members of a unique and conserved family of intracellular
lipid kinases that phosphorylate the 3'-OH group on
phosphatidylinositols or phosphoinositides. The PI3K family
includes kinases with distinct substrate specificities, expression
patterns, and modes of regulation (see, e.g., Katso et al., 2001,
Annu. Rev. Cell Dev. Biol. 17, 615-675; Foster, F. M. et al., 2003,
J Cell Sci 116, 3037-3040). The class I PI3Ks (e.g., p110.alpha.,
p110.beta., p110.gamma., and p110.delta.) are typically activated
by tyrosine kinases or G-protein coupled receptors to generate
PIPS, which engages downstream mediators such as those in the
Akt/PDK1 pathway, mTOR, the Tec family kinases, and the Rho family
GTPases. The class II PI3Ks (e.g., PI3K-C2.alpha., PI3K-C2.beta.,
PI3K-C2.gamma.) and III PI3Ks (e.g., Vps34) play a key role in
intracellular trafficking through the synthesis of PI(3)P and
PI(3,4)P2. Specific exemplary PI3K inhibitors are disclosed
herein.
[0216] The class I PI3Ks comprise a p110 catalytic subunit and a
regulatory adapter subunit. See, e.g., Cantrell, D. A. (2001)
Journal of Cell Science 114: 1439-1445. Four isoforms of the p110
subunit (including PI3K-.alpha. (alpha), PI3K-.beta. (beta),
PI3K-.gamma. (gamma), and PI3K-.delta. (delta) isoforms) have been
implicated in various biological functions. Class I PI3K.alpha. is
involved, for example, in insulin signaling, and has been found to
be mutated in solid tumors. Class I PI3K-.beta. is involved, for
example, in platelet activation and insulin signaling. Class I
PI3K-.gamma. plays a role in mast cell activation, innate immune
function, and immune cell trafficking (chemokines). Class I
PI3K-.delta. is involved, for example, in B-cell and T-cell
activation and function and in Fc receptor signaling in mast cells.
In some embodiments provided herein, the PI3K inhibitor is a class
I PI3K inhibitor. In some such embodiments, the PI3K inhibitor
inhibits a PI3K-.alpha. (alpha), PI3K-.beta. (beta), PI3K-.gamma.
(gamma), or PI3K-.delta. (delta) isoform, or a combination
thereof.
[0217] Downstream mediators of PI3K signal transduction include Akt
and mammalian target of rapamycin (mTOR). Akt possesses a
pleckstrin homology (PH) domain that binds PIPS, leading to Akt
kinase activation. Akt phosphorylates many substrates and is a
central downstream effector of PI3K for diverse cellular responses.
One important function of Akt is to augment the activity of mTOR,
through phosphorylation of TSC2 and other mechanisms. mTOR is a
serine-threonine kinase related to the lipid kinases of the PI3K
family.
2. Compositions and Methods
[0218] B-cell lymphoma 2 (Bcl-2) is a family of proteins that
regulates cell death, e.g., by controlling the regulation of
mitochondrial membrane and the release of cytochrome c from
mitochondria during apoptosis. Bcl-2 plays an important survival
role for many cancers, and the overexpression of the Bcl-2 proteins
has been associated with tumor initiation, progression, and
resistance to certain anticancer therapies.
[0219] Damage to the Bcl-2 gene has been identified as a cause of a
number of cancers, including melanoma, breast, prostate, chronic
lymphocytic leukemia, and lung cancer, and a possible cause of
schizophrenia and autoimmunity. It is also a cause of resistance to
cancer treatments. Over-expression of anti-apoptotic Bcl-2 members
such as Bcl-2, Bcl-xL, and Mcl-1, occurs frequently in cancers,
particularly hematological malignancies such as acute myelogenous
leukemia (AML), resulting in defective apoptosis leading to
enhanced cell survival and drug resistance. Letai A, et al., Cancer
Cell 2004; 6:241-9; Oltersdorf T, et al., Nature 2005; 435:677-81;
and Letai A., J Clin Invest 2005; 115:2648-55. Several agents have
been developed to target these proteins directly, e.g., ABT-737, a
BH3 mimetic that binds with high affinity to and antagonizes the
functions of Bcl-2 and Bcl-xL but not Mcl-1. Merino et al. Blood
2012 (119) 24; 5807-5816. Preclinical studies demonstrated that
ABT-737 induces apoptosis and potentiates the anti-tumor activity
of multiple agents in various cancers, including leukemia. Vo, T.
T., et al., Cell 151, 344-355 (2012); and Davids, M. S., et al.,
Blood (2012).
[0220] ABT-263, a clinical derivative of ABT-737 is currently
undergoing phase I and II clinical evaluation in various tumor
types including leukemia.
[0221] There is a need for an effective and safe combination
therapy of a PI3K inhibitor and a Bcl-2 inhibitor for treating
cancers.
[0222] Without being bound by theory, this disclosure provides
experiments indicating that treating a patient with a PI3K
inhibitor induces an apoptotic cascade characterized by
up-regulation of the pro-apoptotic factors BMF, BIK, Bcl2L11 (also
called BIM), PMAIP1 (also called NOXA), BBC3 (also called PUMA),
and HRK (see Example 2). These factors are upstream of, and
negatively regulate, the anti-apoptotic factor Bcl-2. This
disclosure also provides that Bcl-2 levels rise in some patients
treated with a PI3K inhibitor (see Example 2). Taken together,
these data indicate that elevated levels of Bcl-2 can prevent the
pro-apoptotic factors from successfully initiating apoptosis. These
results suggest that inhibiting Bcl-2 will unblock the apoptotic
cascade that was initiated by the PI3K inhibitor. Furthermore,
experiments in cell lines resistant to a PI3K inhibitor indicate
that pro-apoptotic genes promote responsiveness to a PI3K inhibitor
(see Example 3). These experiments suggest that a combination
therapy with a PI3K-inhibitor and a Bcl-2 inhibitor can produce
unexpectedly high efficacy by acting in concert with high levels of
pro-apoptotic signaling factors, to trigger apoptosis. Bcl
[0223] Bcl The experimental results suggest certain therapeutic
regimens. For example, according to the non-limiting theory herein,
a PI3K inhibitor can be administered to a patient (e.g., as a
monotherapy) until rising Bcl-2 levels reduce its efficacy. At that
point, a Bcl-2 inhibitor can be administered to counteract the
resistance mechanism. In other embodiments, a PI3K inhibitor and a
Bcl-2 inhibitor can be administered over the same period, so that
Bcl-2 activity can be kept low, thereby preventing resistance from
developing. As an additional example, the observation that PI3K
inhibitors and Bcl-2 inhibitor show synergistic efficacy indicates
that administering both drugs at normal (e.g., monotherapy) levels
produces greater efficacy than either agent alone. As still another
example, the observation that PI3K inhibitors and Bcl-2 inhibitor
show synergistic efficacy suggests that one can lower the dose,
frequency, or dose and frequency of one or both of the inhibitors,
and still maintain efficacy. Lowering the dose and/or frequency of
administration can lower side effects.
[0224] In certain embodiments, provided herein are pharmaceutical
compositions comprising a PI3K inhibitor, or a pharmaceutically
acceptable form thereof, in combination with a Bcl-2 inhibitor, or
a pharmaceutically acceptable form thereof. In certain embodiments,
the combination is in a therapeutically effective amount. In
certain embodiments, the combination is in a synergistically
therapeutically effective amount. In certain embodiments, the
combination is synergistic. In certain embodiments, the combination
has a synergistic effect. In certain embodiments, the combination
has a synergistic anti-cancer effect. In certain embodiments, the
combination has a synergistic therapeutic effect.
[0225] Also provided herein are methods of treating (e.g.,
inhibiting, managing, or preventing) a cancer in a subject
comprising administering to the subject a PI3K inhibitor, or a
pharmaceutically acceptable form thereof, in combination with a
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof. In
certain embodiments, the combination is synergistic. In certain
embodiments, the combination has a synergistic effect. In certain
embodiments, the combination has a synergistic anti-cancer effect.
In certain embodiments, the combination has a synergistic
therapeutic effect. In certain embodiments, the combination is
administered in a therapeutically effective amount. In certain
embodiments, the combination is administered in a synergistically
therapeutically effective amount.
[0226] In certain embodiments, the compositions and methods
provided herein are utilized where a monotherapy of one of the
therapeutic agents is becoming less effective due to drug
resistance or where the relatively high dosage of monotherapy lead
to undesirable side effects.
[0227] Without being limited by a particular theory, recent studies
indicate that PI3K inhibitors down-regulate Mcl-1, an event that
plays an important role in transformed cell lethality. Balakrishnan
et al. (ASH Annual Meeting). 2013; No. 4167. Furthermore, Mcl-1, as
well as Bim, which is also regulated by the PI3K pathway, play
important roles in determining ABT-737 sensitivity. Merino et al.
Blood 2012 (119) 24; 5807-5816. In addition, PI3K activity can lead
to inhibition of apoptosis and therefore antagonize the effects of
Bcl-2 inhibitors. Michalak et al., BBRC (2005) 331:786-798; Duronio
et al., Biochem. J. (2008) 415: 333-344; Franke et al., Oncogene
(2003) 22: 8983-8998; Wu et al., Oncogene (2001) 20: 240-251; and
Srinivasan et al Cell (2009) 139: 573-586.
[0228] These considerations, together with the role of Bcl-2 and
Bcl-xL dysregulation in leukemogenesis, raise the possibility that
interference with Bcl-2 and Bcl-xL function can be compatible with
inhibition of PI3 kinase activity. It has been surprisingly found
that the PI3K inhibitors and Bcl-2 inhibitors combinations provided
herein exhibit synergistic actions in treating various types of
cancers.
[0229] PI3K inhibitors that can be used in the compositions and
methods provided herein include, but are not limited to, those
described in, e.g., WO 09/088990, WO 09/088086, WO 2011/008302, WO
2010/036380, WO 2010/006086, WO 09/114870, WO 05/113556,
WO2014072937, WO2014071125; US 2009/0312310, and US 2011/0046165,
the entirety of each incorporated herein by reference. Additional
PI3K inhibitors that can be used in the compositions and methods
provided herein include, but are not limited to, AMG-319, GSK
2126458
(2,4-Difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyrid-
inyl}benzenesulfonamide), GSK 1059615
(5Z-[[4-(4-pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidinedione),
GDC-0032
(4-[5,6-dihydro-2-[3-methyl-1-(1-methylethyl)-1H-1,2,4-triazol-5-
-yl]imidazo[1,2-d][1,4]benzoxazepin-9-yl]-.alpha.,.alpha.-dimethyl-1H-Pyra-
zole-1-acetamide), GDC-0980
((S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]p-
yrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one),
GDC-0941
(2-(1H-indazol-4-yl)-6-((4-(methylsulfonyl)piperazin-1-yl)methyl)-4-morph-
olinothieno[3,2-d]pyrimidine), XL147
(N-(3-(benzo[c][1,2,5]thiadiazol-5-ylamino)quinoxalin-2-yl)-4-methylbenze-
nesulfonamide), XL499, XL765 (SAR245409,
N-[4-[[[3-[(3,5-dimethoxyphenyl)amino]-2-quinoxalinyl]amino]sulfonyl]phen-
yl]-3-methoxy-4-methyl-benzamide), PF-4691502
(2-amino-6-(6-methoxypyridin-3-yl)-4-methyl-8-[(1R,4R)-4-(2-hydroxyethoxy-
l)cyclohexyl]-7H,8H-pyrido[2,3-d]pyrimidin-7-one), BKM 120
(buparlisib,
5-(2,6-dimorpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine),
Idelalisib (CAL-101, GS1101,
(S)-2-(1-(9H-purin-6-ylamino)propyl)-5-fluoro-3-phenylquinazolin-4(3H)-on-
e), CAL 263, SF1126
(3-[[2-[[5-[[amino(azaniumyl)methylidene]amino]-2-[[4-oxo-4-[4-(4-oxo-8-p-
henylchromen-2-yl)morpholin-4-ium-4-yl]oxybutanoyl]amino]pentanoyl]amino]a-
cetyl]amino]-4-(1-carboxylatopropylamino)-4-oxobutanoate), PX-866
(sonolisib,
[(3aR,6E,9S,9aR,10R,11aS)-6-[[bis(prop-2-enyl)amino]methylidene]-5-hydrox-
y-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxo-2,3,3a,9,10,11-hexahydroi-
ndeno[4,5-h]isochromen-10-yl]acetate), BEZ235
(2-methyl-2-(4-(3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c-
]quinolin-1-yl)phenyl)propanenitrile), GS9820 (CAL-120,
(S)-2-(1-((9H-purin-6-yl)amino)ethyl)-6-fluoro-3-phenylquinazolin-4(3H)-o-
ne), BYL719 ((2S)-1,2-Pyrrolidinedicarboxamide,
N1-[4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethylethyl)-4-pyridinyl]-2-thia-
zolyl]), RP6503, RP6530, TGR1202
(((S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyr-
imidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one)),
INK1117 (MLN-1117), PX-866, BAY 80-6946
(2-amino-N-(7-methoxy-8-(3-morpholinopropoxy)-2,3-dihydroimidazo[1,2-c]qu-
inazolin-5-yl)pyrimidine-5-carboxamide), IC87114
(2-((6-amino-9H-purin-9-yl)methyl)-5-methyl-3-o-tolylquinazolin-4(3H)-one-
), Palomid 529
(3-(4-methoxybenzyloxy)-8-(1-hydroxyethyl)-2-methoxy-6H-benzo[c]chromen-6-
-one), ZSTK474
(2-(difluoromethyl)-1-(4,6-dimorpholino-1,3,5-triazin-2-yl)-1H-benzo[d]im-
idazole), PWT33597, TG100-115
(6,7-Bis(3-hydroxyphenyl)pteridine-2,4-diamine), GNE-477
(5-[7-methyl-4-(morpholin-4-yl)-6-[(4-methylsulfonylpiperazin-1-yl)methyl-
]thieno[3,2-d]pyrimidin-2-yl]pyrimidin-2-amine), CUDC-907
(N-hydroxy-2-(((2-(6-methoxypyridin-3-yl)-4-morpholinothieno[3,2-d]pyrimi-
din-6-yl)methyl)(methyl)amino)pyrimidine-5-carboxamide), AEZS-136,
BGT-226
(8-(6-methoxypyridin-3-yl)-3-methyl-1-(4-(piperazin-1-yl)-3-(trifluoromet-
hyl)phenyl)-1H-imidazo[4,5-c]quinolin-2(3H)-one maleic acid),
PF-05212384
(1-(4-(4-(dimethylamino)piperidine-1-carbonyl)phenyl)-3-(4-(4,6-dimorphol-
ino-1,3,5-triazin-2-yl)phenyl)urea), LY3023414, PI-103
(3-[4-(4-morpholinyl)pyrido[3',2':4,5]furo[3,2-d]pyrimidin-2-yl]-phenol),
INCB040093, CAL-130
((S)-2-(1-((2-amino-9H-purin-6-yl)amino)ethyl)-5-methyl-3-(o-tolyl)quinaz-
olin-4(3H)-one), LY294002 (2-Morpholin-4-yl-8-phenylchromen-4-one)
and wortmannin.
[0230] In one embodiment, the PI3K inhibitor is Idelalisib
(GS1101), CAL-130, BKM 120, GDC-0941, PX-866, GDC-0032, BAY
80-6946, BEZ235, BYL719, BGT-226, PF-4691502, GDC-0980, GSK
2126458, PF-05212384, XL765, or XL147.
[0231] In certain embodiments, a PI3K inhibitor is a compound that
inhibits one or more PI3K isoforms, e.g., alpha, beta, delta, or
gamma isoform. In one embodiment, a PI3K inhibitor is a compound
that inhibits one or more PI3K isoforms with an IC.sub.50 of less
than about 1000 nM, less than about 900 nM, less than about 800 nM,
less than about 700 nM, less than about 600 nM, less than about 500
nM, less than about 400 nM, less than about 300 nM, less than about
200 nM, less than about 100 nM, less than about 75 nM, less than
about 50 nM, less than about 25 nM, less than about 20 nM, less
than about 15 nM, less than about 10 nM, less than about 10 nM,
less than about 5 nM, or less than about 1 nM.
[0232] In one embodiment, the PI3K inhibitor is a compound that
inhibits alpha, beta, delta and gamma isoforms. In another
embodiment, the PI3K inhibitor is a compound that inhibits beta,
delta, and gamma isoforms. In another embodiment, the PI3K
inhibitor is a compound that inhibits the delta and gamma
isoforms.
[0233] In certain embodiments, the PI3K inhibitor is a PI3K isoform
selective inhibitor. In one embodiment, the PI3K inhibitor is a
PI3K alpha selective inhibitor. In another embodiment, the PI3K
inhibitor is a PI3K beta selective inhibitor.
[0234] In certain embodiments, the PI3K inhibitor is a PI3K delta
selective inhibitor. In one embodiment, the PI3K delta selective
inhibitor selectively inhibits PI3K delta isoform over PI3K gamma
isoform. In one embodiment, the PI3K delta selective inhibitor has
a gamma/delta selectivity ratio of greater than 1, greater than
about 5, greater than about 10, greater than about 50, greater than
about 100, greater than about 200, greater than about 400, greater
than about 600, greater than about 800, greater than about 1000,
greater than about 1500, greater than about 2000, greater than
about 5000, greater than about 10,000, or greater than about
20,000. In one embodiment, the PI3K delta selective inhibitor has a
gamma/delta selectivity ratio in the range of from greater than 1
to about 5, from about 5 to about 10, from about 10 to about 50,
from about 50 to about 850, or greater than about 850. In one
embodiment, the gamma/delta selectivity ratio is determined by
dividing the inhibitor's IC.sub.50 against PI3K gamma isoform by
the inhibitor's IC.sub.50 against PI3K delta isoform.
[0235] In certain embodiments, the PI3K inhibitor is a PI3K delta
selective inhibitor. In one embodiment, the PI3K delta selective
inhibitor selectively inhibits PI3K delta isoform over PI3K alpha
isoform. In one embodiment, the PI3K delta selective inhibitor has
an alpha/delta selectivity ratio of greater than 1, greater than
about 5, greater than about 10, greater than about 50, greater than
about 100, greater than about 200, greater than about 400, greater
than about 600, greater than about 800, greater than about 1000,
greater than about 1500, greater than about 2000, greater than
about 5000, greater than about 10,000, or greater than about
20,000. In one embodiment, the PI3K delta selective inhibitor has
an alpha/delta selectivity ratio in the range of from greater than
1 to about 5, from about 5 to about 10, from about 10 to about 50,
from about 50 to about 850, or greater than about 850. In one
embodiment, the alpha/delta selectivity ratio is determined by
dividing the inhibitor's IC.sub.50 against PI3K alpha isoform by
the inhibitor's IC.sub.50 against PI3K delta isoform.
[0236] In certain embodiments, the PI3K inhibitor is a PI3K delta
selective inhibitor. In one embodiment, the PI3K delta selective
inhibitor selectively inhibits PI3K delta isoform over PI3K beta
isoform. In one embodiment, the PI3K delta selective inhibitor has
a beta/delta selectivity ratio of greater than 1, greater than
about 5, greater than about 10, greater than about 50, greater than
about 100, greater than about 200, greater than about 400, greater
than about 600, greater than about 800, greater than about 1000,
greater than about 1500, greater than about 2000, greater than
about 5000, greater than about 10,000, or greater than about
20,000. In one embodiment, the PI3K delta selective inhibitor has a
beta/delta selectivity ratio in the range of from greater than 1 to
about 5, from about 5 to about 10, from about 10 to about 50, from
about 50 to about 850, or greater than about 850. In one
embodiment, the beta/delta selectivity ratio is determined by
dividing the inhibitor's IC.sub.50 against PI3K beta isoform by the
inhibitor's IC.sub.50 against PI3K delta isoform.
[0237] In certain embodiments, the PI3K inhibitor is selective for
both gamma and delta. In one embodiment, the PI3K gamma and delta
selective inhibitor selectively inhibits PI3K gamma and delta
isoforms over PI3K beta isoform. In one embodiment, the PI3K gamma
and delta selective inhibitor has a beta/delta selectivity ratio of
greater than 1, greater than about 5, greater than about 10,
greater than about 50, greater than about 100, greater than about
200, greater than about 400, greater than about 600, greater than
about 800, greater than about 1000, greater than about 1500,
greater than about 2000, greater than about 5000, greater than
about 10,000, or greater than about 20,000 and a beta/gamma
selectivity ratio of greater than 1, greater than about 5, greater
than about 10, greater than about 50, greater than about 100,
greater than about 200, greater than about 400, greater than about
600, greater than about 800, greater than about 1000, greater than
about 1500, greater than about 2000, greater than about 5000,
greater than about 10,000, or greater than about 20,000. In one
embodiment, the PI3K delta selective inhibitor has a beta/delta
selectivity ratio in the range of from greater than 1 to about 5,
from about 5 to about 10, from about 10 to about 50, from about 50
to about 850, or greater than about 850 and a beta/gamma
selectivity ratio in the range of from greater than 1 to about 5,
from about 5 to about 10, from about 10 to about 50, from about 50
to about 850, or greater than about 850. In one embodiment, the
beta/delta selectivity ratio is determined by dividing the
inhibitor's IC.sub.50 against PI3K beta isoform by the inhibitor's
IC.sub.50 against PI3K delta isoform and the beta/gamma selectivity
ratio is determined by dividing the inhibitor's IC.sub.50 against
PI3K beta isoform by the inhibitor's IC.sub.50 against PI3K gamma
isoform.
[0238] PI3K delta inhibitors that can be used in the compositions
and methods provided herein include, but are not limited to,
GSK-2269557
(2-(6-(1H-indol-4-yl)-1H-indazol-4-yl)-5-((4-isopropylpiperazin-1-yl)meth-
yl)oxazole), GS-9820, GS-1101
(5-fluoro-3-phenyl-2-([S)]-1-[9H-purin-6-ylamino]-propyl)-3H-quinazolin-4-
-one), AMG319, or TGR-1202
(((S)-2-(1-(4-amino-3-(3-fluoro-4-isopropoxyphenyl)-1H-pyrazolo[3,4-d]pyr-
imidin-1-yl)ethyl)-6-fluoro-3-(3-fluorophenyl)-4H-chromen-4-one)),
or a mixture thereof. In one embodiment, the PI3K delta inhibitor
is GS1101.
[0239] In one embodiment, the PI3K inhibitor is a PI3K inhibitor as
described in WO 2005/113556, the entirety of which is incorporated
herein by reference. In one embodiment, the PI3K inhibitor is
Compound Nos. 113 or 107 as described in WO2005/113556.
[0240] In one embodiment, the PI3K inhibitor is a PI3K inhibitor as
described in WO2014/006572, the entirety of which is incorporated
herein by reference. In one embodiment, the PI3K inhibitor is
Compound Nos. A1, A2, B, B1, or B2 as described in
WO2014/006572.
[0241] In certain embodiments, the PI3K inhibitor is a PI3K
delta/gamma dual inhibitor. In one embodiment, the PI3K delta/gamma
dual inhibitor has an IC.sub.50 value against PI3K alpha that is at
least 5.times., 10.times., 20.times., 50.times., 100.times.,
200.times., 500.times., or 1000.times. higher than its IC.sub.50
values against delta and gamma.
[0242] In certain embodiments, the PI3K inhibitor is Compound 1A of
the structure:
##STR00002##
or a pharmaceutically acceptable form thereof.
[0243] In certain embodiments, the PI3K inhibitor is Compound 1 of
the structure:
##STR00003##
or a pharmaceutically acceptable form thereof.
[0244] Compound 1A has a chemical name of
3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)-one.
Compound 1 has a chemical name of
(S)-3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)--
one. An exemplary method for synthesizing Compound 1A and Compound
1 has been previously described in U.S. Pat. No. 8,193,182, which
is incorporated by reference in its entirety. Compound 1 is a
PI3K-.delta., -.gamma. inhibitor and can be used to treat cancers.
See U.S. Pat. No. 8,193,182.
[0245] Compound 1 provided herein contains one chiral center, and
can exist as a mixture of enantiomers, e.g., a racemic mixture.
This application encompasses the use of stereomerically pure forms
of such a compound, as well as the use of mixtures of those forms.
For example, mixtures comprising equal or unequal amounts of the
enantiomers of Compound 1 provided herein may be used in methods
and compositions disclosed herein. These isomers may be
asymmetrically synthesized or resolved using standard techniques
such as chiral columns or chiral resolving agents. See, e.g.,
Jacques, J., et al., Enantiomers, Racemates and Resolutions
(Wiley-Interscience, New York, 1981); Wilen, S. H., et al.,
Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of Carbon
Compounds (McGraw-Hill, NY, 1962); and Wilen, S. H., Tables of
Resolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed.,
Univ. of Notre Dame Press, Notre Dame, Ind., 1972).
[0246] In one embodiment, the PI3K inhibitor provided herein is a
mixture of Compound 1 and its (R)-enantiomer. In one embodiment,
the PI3K inhibitor provided herein is a racemic mixture of Compound
1 and its (R)-enantiomer. In other embodiments, the compound
mixture has an (S)-enantiomeric purity of greater than about 55%,
about 60%, about 65%, about 70%, about 75%, about 80%, about 85%,
about 90%, about 95%, about 96%, about 97%, about 98%, about 99%,
about 99.5%, or more. In other embodiments, the compound mixture
has an (S)-enantiomeric purity of greater than about 55% to about
99.5%, greater than about 60% to about 99.5%, greater than about
65% to about 99.5%, greater than about 70% to about 99.5%, greater
than about 75% to about 99.5%, greater than about 80% to about
99.5%, greater than about 85% to about 99.5%, greater than about
90% to about 99.5%, greater than about 95% to about 99.5%, greater
than about 96% to about 99.5%, greater than about 97% to about
99.5%, greater than about 98% to greater than about 99.5%, greater
than about 99% to about 99.5%, or more.
[0247] In other embodiments, the compound mixture has an
(R)-enantiomeric purity of greater than about 55%, about 60%, about
65%, about 70%, about 75%, about 80%, about 85%, about 90%, about
95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, or
more. In other embodiments, the compound mixture has an
(R)-enantiomeric purity of greater than about 55% to about 99.5%,
greater than about 60% to about 99.5%, greater than about 65% to
about 99.5%, greater than about 70% to about 99.5%, greater than
about 75% to about 99.5%, greater than about 80% to about 99.5%,
greater than about 85% to about 99.5%, greater than about 90% to
about 99.5%, greater than about 95% to about 99.5%, greater than
about 96% to about 99.5%, greater than about 97% to about 99.5%,
greater than about 98% to greater than about 99.5%, greater than
about 99% to about 99.5%, or more.
[0248] As used herein, Compound 1 also refers to any crystal form
or polymorph of
(S)-3-(1-((9H-purin-6-yl)amino)ethyl)-8-chloro-2-phenylisoquinolin-1(2H)--
one. In some embodiments, a polymorph of Compound 1, or a
pharmaceutically form thereof, disclosed herein is used. Exemplary
polymorphs are disclosed in U.S. Patent Publication No.
2012/0184568, which is hereby incorporated by reference in its
entirety. In one embodiment, the compound is Form A of Compound 1.
In one embodiment, the compound is Form B of Compound 1. In one
embodiment, the compound is Form C of Compound 1. In one
embodiment, the compound is Form D of Compound 1. In one
embodiment, the compound is Form E of Compound 1. In one
embodiment, the compound is Form F of Compound 1. In one
embodiment, the compound is Form G of Compound 1. In one
embodiment, the compound is Form H of Compound 1. In one
embodiment, the compound is Form I of Compound 1. In one
embodiment, the compound is Form J of Compound 1. In one
embodiment, the compound is a mixture of solid forms (e.g.,
polymorphs and/or amorphous forms) of Compound 1 disclosed
herein.
[0249] Any of the compounds disclosed herein can be in the form of
pharmaceutically acceptable salts, hydrates, solvates, chelates,
non-covalent complexes, isomers, prodrugs, isotopically labeled
derivatives, or mixtures thereof.
[0250] Bcl-2 inhibitors that can be used in the compositions and
methods provided herein include, but are not limited to ABT-199,
ABT-263, ABT-737, G3139 (genasense or oblimersen), GX15-070
(obatoclax mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101),
antimycin A, and apogossypol. In one embodiment, Bcl-2 inhibitors
that can be used in the compositions and methods provided herein
include, but are not limited to, those described in, e.g.,
WO2006/009869, WO2007075387, WO2008024337, WO2008060569, or
WO2013110890, the entirety of each incorporated herein by
reference.
[0251] ABT-737 has the chemical name of
(4-[4-[(4'-chloro[1,1'-biphenyl]-2-yl)methyl]-1-piperazinyl]-N-[[4-[[(1R)-
-3-(dimethylamino)-1-[(phenylthio)methyl]propyl]amino]-3-nitrophenyl]sulfo-
nyl]-benzamide). Bruncko et al. (2007) J. Med. Chem. 50:641-662;
U.S. Patent Application Publication No. 2007/0072860.
[0252] G3139 (genasense or oblimersen) is an 18-mer
phosphorothioate oligodeoxyribonucleotide. R. J. Klasa, et al.
(2002) Antisense and Nucleic Acid Drug Development 12:193-213.
[0253] GX15-070 (obatoclax mesylate) has a chemical name of
(Z)-2-(5-((3,5-dimethyl-1H-pyrrol-2-yl)methylene)-4-methoxy-5H-pyrrol-2-y-
l)-1H-indole mesylate).
[0254] HA14-1 has a chemical name of ((R)-ethyl
2-amino-6-bromo-4-((R)-1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxy-
late.
[0255] TW-37 has the chemical name of
[4-[[2-(1,1-Dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2--
(1-methylethyl)phenyl]methyl]benzamide).
[0256] Sabutoclax has the chemical name of
((1R)-[2,2'-Binaphthalene]-5,5'-dicarboxamide,
1,1',6,6',7,7'-hexahydroxy-3,3'-dimethyl-N5,N5'-bis[(2R)-2-phenylpropyl])-
.
[0257] Gossypol (AT-101) has the chemical name of
2,2'-bis-(Formyl-1,6,7-trihydroxy-5-isopropyl-3-methylnaphthalene).
[0258] S44563 has the chemical name
(R)-3-((4'-chloro-[1,1'-biphenyl]-2-yl)methyl)-N-((4-(((R)-4-(dimethylami-
no)-1-(phenylthio)butan-2-yl)amino)-3-nitrophenyl)sulfonyl)-2,3,4,4a,5,6-h-
exahydro-1H-pyrazino[1,2-a]quinoline-8-carboxamide.
[0259] Antimycin A has the chemical name of
((2{R},3{S},6{S},7{R},8{R})-3-[(3-formamido-2-hydroxybenzoyl)amino]-8-hex-
yl-2,6-dimethyl-4,9-dioxo-1,5-dioxonan-7-yl 3-methylbutanoate).
[0260] In one embodiment, the Bcl-2 inhibitor is ABT-199. ABT-199
has a chemical name of
4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohex-1-en-1-yl]methyl]piperazin-
-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]amino]phenyl]sulfo-
nyl]-2-[(1H-pyrrolo[2,3-b]pyridin-5-yl)oxy]benzamide, and is of the
structure:
##STR00004##
as described in U.S. Pat. No. 8,546,399, which is incorporated by
reference in its entirety.
[0261] In one embodiment, the Bcl-2 inhibitor is ABT-263. ABT-263
has a chemical name of
(R)-4-(4-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl-
)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amin-
o)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide and is of
the structure:
##STR00005##
as described in U.S. Pat. No. 7,390,799, which is incorporated by
reference in its entirety.
[0262] In certain embodiments, provided herein is a pharmaceutical
composition comprising a therapeutically effective amount of a PI3K
delta selective inhibitor, or a pharmaceutically acceptable form
thereof, and a Bcl-2 inhibitor, or a pharmaceutically acceptable
form thereof. In one embodiment, the PI3K delta selective inhibitor
is GS1101 (CAL-101). In one embodiment, the Bcl-2 inhibitor is
ABT-199, ABT-263, ABT-737, G3139 (genasense or oblimersen),
GX15-070 (obatoclax mesylate), HA14-1, TW-37, sabutoclax, Gossypol
(AT-101), antimycin A, apogossypol, or a mixture thereof. In one
embodiment, the Bcl-2 inhibitor is ABT-199. In another embodiment,
the Bcl-2 inhibitor is ABT-263. In one embodiment, provided herein
is a pharmaceutical composition comprising a therapeutically
effective amount of GS1101, or a pharmaceutically acceptable form
thereof, and ABT-199, or a pharmaceutically acceptable form
thereof. In another embodiment, provided herein is a pharmaceutical
composition comprising a therapeutically effective amount of
GS1101, or a pharmaceutically acceptable form thereof, and ABT-263,
or a pharmaceutically acceptable form thereof.
[0263] In one embodiment of the compositions and methods described
herein, the molar ratio of the PI3K delta inhibitor (e.g., GS1101),
or a pharmaceutically acceptable form thereof, to a Bcl-2 inhibitor
(e.g., ABT-199 or ABT-263), or a pharmaceutically acceptable form
thereof, is in the range of from about 500:1 to about 1:500, from
about 400:1 to about 1:400, from about 300:1 to about 1:300, from
about 200:1 to about 1:200, from about 100:1 to about 1:100, from
about 75:1 to about 1:75, from about 50:1 to about 1:50, from about
40:1 to about 1:40, from about 30:1 to about 1:30, from about 20:1
to about 1:20, from about 10:1 to about 1:10, or from about 5:1 to
about 1:5. In an embodiment, the PI3K delta inhibitor is GS1101 and
the Bcl-2 inhibitor is ABT-199, and the molar ratio of the PI3K
delta inhibitor to the Bcl-2 inhibitor is from about 10:1 to about
1:500, from about 1:1 to about 1:50, from about 1:2 to about 1:10,
from about 1:3 to about 1:5, from about 1:3 to about 1:1, from
about 1:2 to about 1:1, from about 5:1 to about 1:1, from about 2:1
to about 1:1, about 2:1, or about 1.5:1.
[0264] In one embodiment, the PI3K delta inhibitor (e.g., GS1101)
is administered at an amount to reach maximum plasma concentration
at steady state (Cmaxss) at about 1000 ng/mL to about 5000 ng/mL,
about 1000 ng/mL to about 4000 ng/mL, about 1000 ng/mL to about
3000 ng/mL, about 1000 ng/mL to about 2500 ng/mL, about 1400 ng/mL
to about 2300 ng/mL, about 2000 ng/mL to about 2300 ng/mL, or about
2200 ng/mL; and
[0265] the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is
administered at an amount to reach Cmaxss at about 0.1 .mu.g/mL to
about 1000 .mu.g/mL, about 0.1 .mu.g/mL to about 500 .mu.g/mL,
about 0.1 .mu.g/mL to about 250 .mu.g/mL, about 1 .mu.g/mL to about
100 .mu.g/mL, about 1 .mu.g/mL to about 50 .mu.g/mL, about 1
.mu.g/mL to about 25 .mu.g/mL, about 1 .mu.g/mL to about 20
.mu.g/mL, about 1 .mu.g/mL to about 10 .mu.g/mL, about 4 .mu.g/mL,
or about 3 .mu.g/mL.
[0266] In one embodiment, the PI3K delta inhibitor (e.g., GS1101)
is administered at an amount to reach maximum plasma concentration
at steady state (Cmaxss) at less than about 5000 ng/mL, less than
about 4000 ng/mL, less than about 3000 ng/mL, less than about 2000
ng/mL, less than about 1500 ng/mL, less than about 1000 ng/mL, less
than about 500 ng/mL, less than about 100 ng/mL, less than about 50
ng/mL, less than about 25 ng/mL, less than about 10 ng/mL, or less
than about 1 ng/mL.
[0267] In one embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263) is administered at an amount to reach Cmaxss less than
about 1000 .mu.g/mL, less than about 500 .mu.g/mL, less than about
100 .mu.g/mL, less than about 50 .mu.g/mL, less than about 10
.mu.g/mL, less than about 5 .mu.g/mL, less than about 4 .mu.g/mL,
less than about 3 .mu.g/mL, less than about 2 .mu.g/mL, less than
about 1 .mu.g/mL, less than about 0.5 .mu.g/mL, or less than about
0.1 .mu.g/mL.
[0268] In one embodiment, the PI3K delta inhibitor (e.g., GS1101)
is administered at an amount to reach an area under the plasma
concentration-time curve at steady-state (AUCss) at about 5000
ng/mL*hr to about 10000 ng/mL*hr, about 5000 ng/mL*hr to about 9000
ng/mL*hr, about 6000 ng/mL*hr to about 9000 ng/mL*hr, about 6000
ng/mL*hr to about 8000 ng/mL*hr, about 6500 ng/mL*hr to about 7500
ng/mL*hr, or about 7000 ng/mL*hr; and
[0269] the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is
administered at an amount to reach an AUCss at about 0.1 ng/mL*hr
to about 10000 ng/mL*hr, about 1 ng/mL*hr to about 10000 ng/mL*hr,
or about 100 ng/mL*hr to about 5000 ng/mL*hr.
[0270] In one embodiment, the PI3K delta inhibitor (e.g., GS1101)
is administered at an amount to reach an area under the plasma
concentration-time curve at steady-state (AUCss) at less than about
10000 ng/mL*hr, less than about 9500 ng/mL*hr, less than about 9000
ng/mL*hr, less than about 8500 ng/mL*hr, less than about 8000
ng/mL*hr, less than about 7000 ng/mL*hr, less than about 6000
ng/mL*hr, less than about 5000 ng/mL*hr, less than about 4000
ng/mL*hr, less than about 3000 ng/mL*hr, less than about 2000
ng/mL*hr, less than about 1000 ng/mL*hr, less than about 500
ng/mL*hr, less than about 100 ng/mL*hr, less than about 10
ng/mL*hr, or less than about 1 ng/mL*hr.
[0271] In one embodiment, the composition comprises the PI3K delta
inhibitor (e.g., GS1101), or a pharmaceutically acceptable form
thereof, at an amount in the range of from about 0.1 mg to about
500 mg, from about 1 mg to about 500 mg, from about 10 mg to about
500 mg, from about 50 mg to about 500 mg, from about 100 mg to
about 400 mg, from about 200 mg to about 400 mg, from about 250 mg
to about 350 mg, or about 300 mg. In one embodiment, the
composition comprises the PI3K delta inhibitor (e.g., GS1101), or a
pharmaceutically acceptable form thereof, at an amount in the range
of from about 0.1 mg to about 75 mg, from about 1 mg to about 75
mg, from about 5 mg to about 75 mg, from about 5 mg to about 60 mg,
from about 5 mg to about 50 mg, from about 5 mg to about 30 mg,
from about 5 mg to about 25 mg, from about 10 mg to about 25 mg, or
from about 10 mg to about 20 mg.
[0272] In one embodiment, the composition comprises the PI3K delta
inhibitor (e.g., GS1101), or a pharmaceutically acceptable form
thereof, at an amount of less than about 500 mg, less than about
400 mg, less than about 350 mg, less than about 300 mg, less than
about 250 mg, less than about 200 mg, less than about 150 mg, less
than about 100 mg, less than about 75 mg, less than about 50 mg,
less than about 30 mg, less than about 25 mg, less than about 20
mg, less than about 19 mg, less than about 18 mg, less than about
17 mg, less than about 16 mg, less than about 16 mg, less than
about 15 mg, less than about 14 mg, less than about 13 mg, less
than about 12 mg, less than about 11 mg, or less than about 10
mg.
[0273] In certain embodiments, provided herein is a pharmaceutical
composition comprising a therapeutically effective amount of a PI3K
delta/gamma dual inhibitor, or a pharmaceutically acceptable form
thereof, and a Bcl-2 inhibitor, or a pharmaceutically acceptable
form thereof. In one embodiment, the Bcl-2 inhibitor is ABT-199,
ABT-263, ABT-737, G3139 (genasense or oblimersen), GX15-070
(obatoclax mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101),
antimycin A, apogossypol, or a mixture thereof. In one embodiment,
the Bcl-2 inhibitor is ABT-199. In another embodiment, the Bcl-2
inhibitor is ABT-263.
[0274] In one embodiment of the compositions and methods described
herein, the molar ratio of the PI3K delta/gamma dual inhibitor, or
a pharmaceutically acceptable form thereof, to a Bcl-2 inhibitor
(e.g., ABT-199 or ABT-263), or a pharmaceutically acceptable form
thereof, is in the range of from about 500:1 to about 1:500, from
about 400:1 to about 1:400, from about 300:1 to about 1:300, from
about 200:1 to about 1:200, from about 100:1 to about 1:100, from
about 75:1 to about 1:75, from about 50:1 to about 1:50, from about
40:1 to about 1:40, from about 30:1 to about 1:30, from about 20:1
to about 1:20, from about 10:1 to about 1:10, or from about 5:1 to
about 1:5. In an embodiment, the PI3K delta/gamma dual inhibitor is
Compound 1 and the Bcl-2 inhibitor is ABT-199, and the molar ratio
of the PI3K delta/gamma dual inhibitor to the Bcl-2 inhibitor is
from about 10:1 to about 1:100, from about 1:1 to about 1:20, from
about 1:2 to about 1:5, from about 1:3.5 to about 1:4.5, or about
1:4. In an embodiment, the PI3K delta/gamma dual inhibitor is
Compound 1 and the Bcl-2 inhibitor is ABT-263, and the molar ratio
of the PI3K delta/gamma dual inhibitor to the Bcl-2 inhibitor is
from about 10:1 to about 1:100, from about 1:1 to about 1:20, from
about 1:2 to about 1:5, from about 1:3.5 to about 1:4.5, about 1:3,
or about 1:4.
[0275] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount to reach maximum
plasma concentration at steady state (Cmaxss) at about 1000 ng/mL
to about 5000 ng/mL, about 1000 ng/mL to about 4000 ng/mL, about
1000 ng/mL to about 3000 ng/mL, about 1000 ng/mL to about 2500
ng/mL, about 1400 ng/mL to about 2000 ng/mL, about 1400 ng/mL to
about 1500 ng/mL, or about 1487 ng/mL; and
[0276] the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is
administered at an amount to reach Cmaxss at about 0.1 .mu.g/mL to
about 1000 .mu.g/mL, about 0.1 .mu.g/mL to about 500 .mu.g/mL,
about 0.1 .mu.g/mL to about 250 .mu.g/mL, about 1 .mu.g/mL to about
100 .mu.g/mL, about 1 .mu.g/mL to about 50 .mu.g/mL, about 1
.mu.g/mL to about 25 .mu.g/mL, about 1 .mu.g/mL to about 20
.mu.g/mL, about 1 .mu.g/mL to about 10 .mu.g/mL, about 4 .mu.g/mL,
or about 3 .mu.g/mL.
[0277] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount to reach maximum
plasma concentration at steady state (Cmaxss) at less than about
5000 ng/mL, less than about 4000 ng/mL, less than about 3000 ng/mL,
less than about 2000 ng/mL, less than about 1500 ng/mL, less than
about 1000 ng/mL, less than about 500 ng/mL, less than about 100
ng/mL, less than about 50 ng/mL, less than about 25 ng/mL, less
than about 10 ng/mL, or less than about 1 ng/mL.
[0278] In one embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263) is administered at an amount to reach Cmaxss less than
about 1000 .mu.g/mL, less than about 500 .mu.g/mL, less than about
100 .mu.g/mL, less than about 50 .mu.g/mL, less than about 10
.mu.g/mL, less than about 5 .mu.g/mL, less than about 4 .mu.g/mL,
less than about 3 .mu.g/mL, less than about 2 .mu.g/mL, less than
about 1 .mu.g/mL, less than about 0.5 .mu.g/mL, or less than about
0.1 .mu.g/mL.
[0279] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount to reach an area
under the plasma concentration-time curve at steady-state (AUCss)
at about 5000 ng/mL*hr to about 10000 ng/mL*hr, about 5000 ng/mL*hr
to about 9000 ng/mL*hr, about 6000 ng/mL*hr to about 9000 ng/mL*hr,
about 7000 ng/mL*hr to about 9000 ng/mL*hr, about 8000 ng/mL*hr to
about 9000 ng/mL*hr, or about 8787 ng/mL*hr; and
[0280] the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is
administered at an amount to reach an AUCss at about 0.1 ng/mL*hr
to about 10000 ng/mL*hr, about 1 ng/mL*hr to about 10000 ng/mL*hr,
or about 100 ng/mL*hr to about 5000 ng/mL*hr.
[0281] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount to reach an area
under the plasma concentration-time curve at steady-state (AUCss)
at less than about 10000 ng/mL*hr, less than about 9500 ng/mL*hr,
less than about 9000 ng/mL*hr, less than about 8500 ng/mL*hr, less
than about 8000 ng/mL*hr, less than about 7000 ng/mL*hr, less than
about 6000 ng/mL*hr, less than about 5000 ng/mL*hr, less than about
4000 ng/mL*hr, less than about 3000 ng/mL*hr, less than about 2000
ng/mL*hr, less than about 1000 ng/mL*hr, less than about 500
ng/mL*hr, less than about 100 ng/mL*hr, less than about 10
ng/mL*hr, or less than about 1 ng/mL*hr.
[0282] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount that is decreased
by about 1.5 fold to about 50 fold of the amount when administered
alone and the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is
administered at an amount that is decreased by about 1.1 fold to
about 50 fold of the amount when administered alone.
[0283] In one embodiment, the PI3K delta/gamma dual inhibitor
(e.g., Compound 1) is administered at an amount that is decreased
by about 1.5 fold to about 50 fold, about 1.5 fold to about 25
fold, about 1.5 fold to about 20 fold, about 1.5 fold to about 15
fold, about 1.5 fold to about 10 fold, about 2 fold to about 10
fold, about 2 fold to about 8 fold, about 4 fold to about 6 fold,
or about 5 fold of the amount when administered alone; and
the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) is administered at
an amount that is decreased by about 1.1 fold to about 50 fold,
about 1.1 fold to about 40 fold, about 1.1 fold to about 30 fold,
about 1.1 fold to about 25 fold, about 1.1 fold to about 20 fold,
about 1.1 fold to about 15 fold, about 1.1 fold to about 10 fold of
the amount when administered alone.
[0284] In one embodiment, the composition comprises the PI3K
delta/gamma dual inhibitor, or a pharmaceutically acceptable form
thereof, at an amount in the range of from about 0.1 mg to about 75
mg, from about 1 mg to about 75 mg, from about 5 mg to about 75 mg,
from about 5 mg to about 60 mg, from about 5 mg to about 50 mg,
from about 5 mg to about 30 mg, from about 5 mg to about 25 mg,
from about 10 mg to about 25 mg, or from about 10 mg to about 20
mg.
[0285] In one embodiment, the composition comprises the PI3K
delta/gamma dual inhibitor, or a pharmaceutically acceptable form
thereof, at an amount of less than about 25 mg, less than about 20
mg, less than about 19 mg, less than about 18 mg, less than about
17 mg, less than about 16 mg, less than about 16 mg, less than
about 15 mg, less than about 14 mg, less than about 13 mg, less
than about 12 mg, less than about 11 mg, or less than about 10
mg.
[0286] In certain embodiments, provided herein is a pharmaceutical
composition comprising a Compound 1:
##STR00006##
or a pharmaceutically acceptable form thereof, in combination with
a Bcl-2 inhibitor, or a pharmaceutically acceptable form
thereof.
[0287] In some embodiments, the combination is synergistic. In
certain embodiments, the combination has a synergistic effect. In
certain embodiments, the combination has a synergistic anti-cancer
effect. In certain embodiments, the combination has a synergistic
therapeutic effect. In certain embodiments, the combination is in a
therapeutically effective amount. In certain embodiments, the
combination is in a synergistically therapeutically effective
amount.
[0288] In one embodiment, the Bcl-2 inhibitor is ABT-199, ABT-263,
ABT-737, G3139 (genasense or oblimersen), GX15-070 (obatoclax
mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101), antimycin
A, apogossypol, S44563, or a mixture thereof. In one embodiment,
the Bcl-2 inhibitor is ABT-199. In another embodiment, the Bcl-2
inhibitor is ABT-263.
[0289] In certain embodiments, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject Compound 1:
##STR00007##
or a pharmaceutically acceptable form thereof, in combination with
a Bcl-2 inhibitor, or a pharmaceutically acceptable form
thereof.
[0290] In some embodiments, the combination is synergistic. In
certain embodiments, the combination has a synergistic effect. In
certain embodiments, the combination has a synergistic anti-cancer
effect. In certain embodiments, the combination has a synergistic
therapeutic effect. In certain embodiments, the combination is in a
therapeutically effective amount. In certain embodiments, the
combination is in a synergistically therapeutically effective
amount.
[0291] In one embodiment, the Bcl-2 inhibitor is ABT-199, ABT-263,
ABT-737, G3139 (genasense or oblimersen), GX15-070 (obatoclax
mesylate), HA14-1, TW-37, sabutoclax, Gossypol (AT-101), antimycin
A, apogossypol, S44563, or a mixture thereof. In one embodiment,
the Bcl-2 inhibitor is ABT-199. In another embodiment, the Bcl-2
inhibitor is ABT-263.
[0292] In some embodiments of the compositions and methods
described herein, Compound 1, or a pharmaceutically acceptable form
thereof, is used in combination with a Bcl-2 inhibitor (e.g.,
ABT-199 or ABT-263), or a pharmaceutically acceptable form thereof,
at certain molar ratios. In one embodiment, provided herein is a
pharmaceutical composition comprising Compound 1:
##STR00008##
or a pharmaceutically acceptable form thereof, and a Bcl-2
inhibitor, or a pharmaceutically acceptable form thereof, wherein
the molar ratio of Compound 1, or a pharmaceutically acceptable
form thereof, to a Bcl-2 inhibitor (e.g., ABT-199 or ABT-263), or a
pharmaceutically acceptable form thereof, is in the range of from
about 1000:1 to about 1:1000.
[0293] In one embodiment of the compositions and methods described
herein, the molar ratio of Compound 1, or a pharmaceutically
acceptable form thereof, to a Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is in the
range of from about 500:1 to about 1:500, from about 400:1 to about
1:400, from about 300:1 to about 1:300, from about 200:1 to about
1:200, from about 100:1 to about 1:100, from about 75:1 to about
1:75, from about 50:1 to about 1:50, from about 40:1 to about 1:40,
from about 30:1 to about 1:30, from about 20:1 to about 1:20, from
about 10:1 to about 1:10, or from about 5:1 to about 1:5. In some
embodiment, the molar ratio of Compound 1, or a pharmaceutically
acceptable form thereof, to a Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is in the
range of from about 1:3 to about 1:7 or from about 1:5 to about
1:6. In some embodiments, the molar ratio of Compound 1, or a
pharmaceutically acceptable form thereof, to a Bcl-2 inhibitor
(e.g., ABT-199 or ABT-263), or a pharmaceutically acceptable form
thereof, is from about 10:1 to about 1:100, from about 1:1 to about
1:20, from about 1:2 to about 1:5, from about 1:3.5 to about 1:4.5,
about 1:3, or about 1:4. In some embodiment, the molar ratio of
Compound 1, or a pharmaceutically acceptable form thereof, to a
Bcl-2 inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, is about 1:3, about 1:4, about 1:5, about
1:6, or about 1:7.
[0294] In one embodiment of the compositions and methods described
herein, the weight ratio of Compound 1, or a pharmaceutically
acceptable form thereof, to ABT-199, or a pharmaceutically
acceptable form thereof, is in the range of from about 7.5-37.5 of
Compound 1 to from 40-200 of ABT-199. In one embodiment, the weight
ratio is in the range of from about 1:1.1 to about 1:27. In one
embodiment, the weight ratio is in the range of from about 1:2.2 to
about 1:13.5. In one embodiment, the weight ratio is in the range
of from about 1:3.3 to about 1:9.
[0295] In one embodiment of the compositions and methods described
herein, the weight ratio of Compound 1, or a pharmaceutically
acceptable form thereof, to ABT-263, or a pharmaceutically
acceptable form thereof, is in the range of from about 7.5-37.5 of
Compound 1 to from 32.5-162.5 of ABT-263. In one embodiment, the
weight ratio is in the range of from about 1.2:1 to about 1:21. In
one embodiment, the weight ratio is in the range of from about
1:1.7 to about 1:10.5. In one embodiment, the weight ratio is in
the range of from about 1:2.5 to about 1:7.
[0296] In one embodiment, Compound 1 is administered at an amount
to reach maximum plasma concentration at steady state (Cmaxss) at
about 1000 ng/mL to about 5000 ng/mL, about 1000 ng/mL to about
4000 ng/mL, about 1000 ng/mL to about 3000 ng/mL, about 1000 ng/mL
to about 2500 ng/mL, about 1400 ng/mL to about 2000 ng/mL, about
1400 ng/mL to about 1500 ng/mL, or about 1487 ng/mL; and
[0297] ABT-199 or ABT-263 is administered at an amount to reach
Cmaxss at about 0.1 .mu.g/mL to about 1000 .mu.g/mL, about 0.1
.mu.g/mL to about 500 .mu.g/mL, about 0.1 .mu.g/mL to about 250
.mu.g/mL, about 1 .mu.g/mL to about 100 .mu.g/mL, about 1 .mu.g/mL
to about 50 .mu.g/mL, about 1 .mu.g/mL to about 25 .mu.g/mL, about
1 .mu.g/mL to about 20 .mu.g/mL, about 1 .mu.g/mL to about 10
.mu.g/mL, about 4 .mu.g/mL, or about 3 .mu.g/mL.
[0298] In one embodiment, Compound 1 is administered at an amount
to reach an area under the plasma concentration-time curve at
steady-state (AUCss) at about 5000 ng/mL*hr to about 10000
ng/mL*hr, about 5000 ng/mL*hr to about 9000 ng/mL*hr, about 6000
ng/mL*hr to about 9000 ng/mL*hr, about 7000 ng/mL*hr to about 9000
ng/mL*hr, about 8000 ng/mL*hr to about 9000 ng/mL*hr, or about 8787
ng/mL*hr; and
[0299] ABT-199 or ABT-263 is administered at an amount to reach an
AUCss at about 0.1 ng/mL*hr to about 10000 ng/mL*hr, about 1
ng/mL*hr to about 10000 ng/mL*hr, or about 100 ng/mL*hr to about
5000 ng/mL*hr.
[0300] In some embodiments, the composition comprises Compound 1,
or a pharmaceutically acceptable form thereof, and a Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, at certain amounts. In one embodiment, the
composition comprises Compound 1, or a pharmaceutically acceptable
form thereof, at an amount in the range of from about 0.01 mg to
about 75 mg and the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263), or
a pharmaceutically acceptable form thereof, at an amount of in the
range of from about 0.01 mg to about 1100 mg.
[0301] In one embodiment, the composition comprises Compound 1, or
a pharmaceutically acceptable form thereof, at an amount in the
range of from about 0.1 mg to about 75 mg, from about 1 mg to about
75 mg, from about 5 mg to about 75 mg, from about 5 mg to about 60
mg, from about 5 mg to about 50 mg, from about 5 mg to about 30 mg,
from about 5 mg to about 25 mg, from about 10 mg to about 25 mg, or
from about 10 mg to about 20 mg.
[0302] In one embodiment, the composition comprises Compound 1, or
a pharmaceutically acceptable form thereof, at an amount of less
than about 25 mg, less than about 20 mg, less than about 19 mg,
less than about 18 mg, less than about 17 mg, less than about 16
mg, less than about 16 mg, less than about 15 mg, less than about
14 mg, less than about 13 mg, less than about 12 mg, less than
about 11 mg, or less than about 10 mg.
[0303] In one embodiment, the composition comprises the Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, at an amount in the range of from about
0.1 mg to about 800 mg, from about 0.1 mg to about 750 mg, from
about 0.1 mg to about 600 mg, from about 1 mg to about 500 mg, from
about 1 mg to about 400 mg, from about 10 mg to about 300 mg, from
about 50 mg to about 250 mg, from about 50 mg to about 200 mg, from
about 50 mg to about 150 mg, from about 50 mg to about 100 mg, from
about 50 mg to about 90 mg, or from about 50 mg to about 80 mg.
[0304] In one embodiment, the composition comprises the Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, at an amount of less than about 1000 mg,
less than about 800 mg, less than about 750 mg, less than about 500
mg, less than about 400 mg, less than about 350 mg, less than about
300 mg, less than about 250 mg, less than about 200 mg, less than
about 150 mg, less than about 100 mg, less than about 75 mg, less
than about 50 mg, or less than about 25 mg.
[0305] In one embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of less than
about 1000 mg, less than about 800 mg, less than about 750 mg, less
than about 500 mg, less than about 400 mg, less than about 350 mg,
less than about 300 mg, less than about 250 mg, less than about 200
mg, less than about 150 mg, less than about 100 mg, less than about
75 mg, less than about 50 mg, or less than about 25 mg. In one
embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of about 400
mg. In one embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of about 325
mg. In one embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of about 250
mg. In one embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of about 150
mg. In one embodiment, the composition comprises ABT-199, or a
pharmaceutically acceptable form thereof, at an amount of about 100
mg, about 90 mg, about 85 mg, about 80 mg, about 75 mg, about 70
mg, about 65 mg, about 60 mg, about 55 mg, about 50 mg, about 45
mg, about 40 mg, about 35 mg, about 30 mg, about 25 mg, about 20
mg, about 15 mg, about 10 mg, or about 5 mg. In one embodiment, the
composition comprises ABT-199, or a pharmaceutically acceptable
form thereof, at an amount of about 85 mg. In one embodiment, the
composition comprises ABT-199, or a pharmaceutically acceptable
form thereof, at an amount of about 75 mg. In one embodiment, the
composition comprises ABT-199, or a pharmaceutically acceptable
form thereof, at an amount of about 50 mg.
[0306] In one embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of less than
about 1000 mg, less than about 800 mg, less than about 750 mg, less
than about 500 mg, less than about 400 mg, less than about 350 mg,
less than about 300 mg, less than about 250 mg, less than about 200
mg, less than about 150 mg, less than about 100 mg, less than about
75 mg, less than about 50 mg, or less than about 25 mg. In one
embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of about 325
mg. In one embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of about 250
mg. In one embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of about 150
mg. In one embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of about 100
mg, about 90 mg, about 85 mg, about 80 mg, about 75 mg, about 70
mg, about 65 mg, about 60 mg, about 55 mg, about 50 mg, about 45
mg, about 40 mg, about 35 mg, about 30 mg, about 25 mg, about 20
mg, about 15 mg, about 10 mg, or about 5 mg. In one embodiment, the
composition comprises ABT-263, or a pharmaceutically acceptable
form thereof, at an amount of about 85 mg. In one embodiment, the
composition comprises ABT-263, or a pharmaceutically acceptable
form thereof, at an amount of about 75 mg. In one embodiment, the
composition comprises ABT-263, or a pharmaceutically acceptable
form thereof, at an amount of about 50 mg.
[0307] In certain embodiments, the PI3K inhibitor (e.g., Compound 1
or GS1101), or pharmaceutically acceptable form thereof, and the
Bcl-2 inhibitor, or pharmaceutically acceptable form thereof, are
in a single dosage form. In other embodiments, the PI3K inhibitor,
or pharmaceutically acceptable form thereof, and the Bcl-2
inhibitor, or pharmaceutically acceptable form thereof, are in
separate dosage forms.
[0308] In certain embodiments, the PI3K inhibitor (e.g., Compound
1), or pharmaceutically acceptable form thereof, and the Bcl-2
inhibitor, or pharmaceutically acceptable form thereof, are the
only therapeutically active ingredients of the compositions and
methods provided herein. In other embodiments, the compositions and
methods provided herein comprise or use at least one more
therapeutically active ingredient, e.g., a third agent, e.g., an
anti-CD20 antibody.
[0309] In one embodiment, a method described herein further
comprises administration of an anti-CD20 antibody. In one
embodiment, a pharmaceutical composition described herein further
comprises an anti-CD20 antibody. In some such embodiments, the
anti-CD20 antibody is included in the same dosage form as the PI3K
inhibitor and/or second agent. In some such embodiments, the
anti-CD20 antibody is in a separate dosage form as the PI3K
inhibitor and/or second agent. The anti-CD20 antibody can be
administered before, after, or concurrent with the PI3K inhibitor
and/or second agent. In certain embodiments, the anti-CD20 antibody
is selected from one or more of rituximab, ofatumumab and
obinotuzumab.
Cancers
[0310] Subjects that can be treated with a pharmaceutical
composition as provided herein, according to the methods as
provided herein include, but are not limited to, patients that have
been diagnosed as having breast cancer such as a ductal carcinoma,
lobular carcinoma, medullary carcinomas, colloid carcinomas,
tubular carcinomas, and inflammatory breast cancer; ovarian cancer,
including epithelial ovarian tumors such as adenocarcinoma in the
ovary and an adenocarcinoma that has migrated from the ovary into
the abdominal cavity; uterine cancer; cervical cancer such as
adenocarcinoma in the cervix epithelial including squamous cell
carcinoma and adenocarcinomas; prostate cancer, such as a prostate
cancer selected from the following: an adenocarcinoma or an
adenocarcinoma that has migrated to the bone; pancreatic cancer
such as epitheliod carcinoma in the pancreatic duct tissue and an
adenocarcinoma in a pancreatic duct; bladder cancer such as a
transitional cell carcinoma in urinary bladder, urothelial
carcinomas (transitional cell carcinomas), tumors in the urothelial
cells that line the bladder, squamous cell carcinomas,
adenocarcinomas, and small cell cancers; leukemia such as acute
myeloid leukemia (AML), acute lymphocytic leukemia, chronic
lymphocytic leukemia, chronic myeloid leukemia, hairy cell
leukemia, myelodysplasia, myeloproliferative disorders, NK cell
leukemia (e.g., blastic plasmacytoid dendritic cell neoplasm),
acute myelogenous leukemia (AML), chronic myelogenous leukemia
(CML), mastocytosis, chronic lymphocytic leukemia (CLL), multiple
myeloma (MM), and myelodysplastic syndrome (MDS); bone cancer; lung
cancer such as non-small cell lung cancer (NSCLC), which is divided
into squamous cell carcinomas, adenocarcinomas, and large cell
undifferentiated carcinomas, and small cell lung cancer; skin
cancer such as basal cell carcinoma, melanoma, squamous cell
carcinoma and actinic keratosis, which is a skin condition that
sometimes develops into squamous cell carcinoma; eye
retinoblastoma; cutaneous or intraocular (eye) melanoma; primary
liver cancer; kidney cancer; thyroid cancer such as papillary,
follicular, medullary and anaplastic; lymphoma such as diffuse
large B-cell lymphoma, B-cell immunoblastic lymphoma, NK cell
lymphoma (e.g., blastic plasmacytoid dendritic cell neoplasm), and
Burkitt lymphoma; Kaposi's Sarcoma; viral-induced cancers including
hepatitis B virus (HBV), hepatitis C virus (HCV), and
hepatocellular carcinoma; human lymphotropic virus-type 1 (HTLV-1)
and adult T-cell leukemia/lymphoma; and human papilloma virus (HPV)
and cervical cancer; central nervous system cancers (CNS) such as
primary brain tumor, which includes gliomas (astrocytoma,
anaplastic astrocytoma, or glioblastoma multiforme),
oligodendroglioma, ependymoma, meningioma, lymphoma, schwannoma,
and medulloblastoma; peripheral nervous system (PNS) cancers such
as acoustic neuromas and malignant peripheral nerve sheath tumor
(MPNST) including neurofibromas and schwannomas, malignant
fibrocytoma, malignant fibrous histiocytoma, malignant meningioma,
malignant mesothelioma, and malignant mixed MUllerian tumor; oral
cavity and oropharyngeal cancers such as, hypopharyngeal cancer,
laryngeal cancer, nasopharyngeal cancer, and oropharyngeal cancer;
stomach cancers such as lymphomas, gastric stromal tumors, and
carcinoid tumors; testicular cancers such as germ cell tumors
(GCTs), which include seminomas and nonseminomas, and gonadal
stromal tumors, which include Leydig cell tumors and Sertoli cell
tumors; thymus cancer such as to thymomas, thymic carcinomas,
Hodgkin lymphoma, non-Hodgkin lymphomas carcinoids or carcinoid
tumors; rectal cancer; and colon cancer.
[0311] In one embodiment, the cancer or disease that may be treated
(e.g., inhibited or prevented) by methods, compositions, or kits
provided herein includes a blood disorder or a hematologic
malignancy. In one embodiment, the cancer or disease that may be
treated is an acoustic neuroma, adenocarcinoma, adrenal gland
cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma,
lymphangioendotheliosarcoma, hemangiosarcoma), benign monoclonal
gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder
cancer, breast cancer (e.g., adenocarcinoma of the breast,
papillary carcinoma of the breast, mammary cancer, medullary
carcinoma of the breast), brain cancer (e.g., meningioma; glioma,
e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchus
cancer, cervical cancer (e.g., cervical adenocarcinoma),
choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer
(e.g., colon cancer, rectal cancer, colorectal adenocarcinoma),
epithelial carcinoma, ependymoma, endotheliosarcoma (e.g., Kaposi's
sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial
cancer, esophageal cancer (e.g., adenocarcinoma of the esophagus,
Barrett's adenocarinoma), Ewing sarcoma, familiar
hypereosinophilia, gastric cancer (e.g., stomach adenocarcinoma),
gastrointestinal stromal tumor (GIST), head and neck cancer (e.g.,
head and neck squamous cell carcinoma, oral cancer (e.g., oral
squamous cell carcinoma (OSCC)), heavy chain disease (e.g., alpha
chain disease, gamma chain disease, mu chain disease),
hemangioblastoma, inflammatory myofibroblastic tumors, immunocytic
amyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms'
tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular
cancer (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic
carcinoma, small cell lung cancer (SCLC), non-small cell lung
cancer (NSCLC), adenocarcinoma of the lung), leukemia (e.g., acute
lymphocytic leukemia (ALL), which includes B-lineage ALL and
T-lineage ALL, chronic lymphocytic leukemia (CLL), prolymphocytic
leukemia (PLL), hairy cell leukemia (HCL) and Waldenstrom's
macroglobulinemia (WM); peripheral T cell lymphomas (PTCL), adult T
cell leukemia/lymphoma (ATL), cutaneous T-cell lymphoma (CTCL),
large granular lymphocytic leukemia (LGF), Hodgkin's disease and
Reed-Stemberg disease; acute myelocytic leukemia (AML), chronic
myelocytic leukemia (CML), chronic lymphocytic leukemia (CLL)),
lymphoma (e.g., Hodgkin lymphoma (HL), non-Hodgkin lymphoma (NHL),
follicular lymphoma, diffuse large B-cell lymphoma (DLBCL), mantle
cell lymphoma (MCL)), leiomyosarcoma (LMS), mastocytosis (e.g.,
systemic mastocytosis), multiple myeloma (MM), myelodysplastic
syndrome (MDS), mesothelioma, myeloproliferative disorder (MPD)
(e.g., polycythemia Vera (PV), essential thrombocytosis (ET),
agnogenic myeloid metaplasia (AMM) a.k.a. myelofibrosis (MF),
chronic idiopathic myelofibrosis, chronic myelocytic leukemia
(CML), chronic neutrophilic leukemia (CNL), hypereosinophilic
syndrome (HES)), neuroblastoma, neurofibroma (e.g.,
neurofibromatosis (NF) type 1 or type 2, schwannomatosis),
neuroendocrine cancer (e.g., gastroenteropancreatic neuroendoctrine
tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer
(e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian
adenocarcinoma), Paget's disease of the vulva, Paget's disease of
the penis, papillary adenocarcinoma, pancreatic cancer (e.g.,
pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm
(IPMN)), pinealoma, primitive neuroectodermal tumor (PNT), prostate
cancer (e.g., prostate adenocarcinoma), rhabdomyosarcoma,
retinoblastoma, salivary gland cancer, skin cancer (e.g., squamous
cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell
carcinoma (BCC)), small bowel cancer (e.g., appendix cancer), soft
tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH),
liposarcoma, malignant peripheral nerve sheath tumor (MPNST),
chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland
carcinoma, sweat gland carcinoma, synovioma, testicular cancer
(e.g., seminoma, testicular embryonal carcinoma), thyroid cancer
(e.g., papillary carcinoma of the thyroid, papillary thyroid
carcinoma (PTC), medullary thyroid cancer), and Waldenstrom's
macroglobulinemia.
[0312] In one embodiment, the cancer or disease being treated or
prevented, such as a blood disorder or hematologic malignancy, has
a high expression level of one or more PI3K isoform(s) (e.g.,
PI3K-.alpha., PI3K-.beta., PI3K-.delta., or PI3K-.gamma., or a
combination thereof).
[0313] In one embodiment, the cancer or disease that may be treated
or prevented by methods, compositions, or kits provided herein
includes a blood disorder or a hematologic malignancy, including,
but not limited to, myeloid disorder, lymphoid disorder, leukemia,
lymphoma, myelodysplastic syndrome (MDS), myeloproliferative
disease (MPD), mast cell disorder, and myeloma (e.g., multiple
myeloma), among others.
[0314] In one embodiment, the blood disorder or the hematologic
malignancy includes, but is not limited to, acute lymphoblastic
leukemia (ALL), T-cell ALL (T-ALL), B-cell ALL (B-ALL), acute
myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic
myelogenous leukemia (CML), blast phase CML, small lymphocytic
lymphoma (SLL), CLL/SLL, blast phase CLL, Hodgkin lymphoma (HL),
non-Hodgkin lymphoma (NHL), B-cell NHL, T-cell NHL, indolent NHL
(iNHL), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma
(MCL), aggressive B-cell NHL, B-cell lymphoma (BCL), Richter's
syndrome (RS), T-cell lymphoma (TCL), peripheral T-cell lymphoma
(PTCL), cutaneous T-cell lymphoma (CTCL), transformed mycosis
fungoides, Sezary syndrome, anaplastic large-cell lymphoma (ALCL),
follicular lymphoma (FL), Waldenstrom macroglobulinemia (WM),
lymphoplasmacytic lymphoma, Burkitt lymphoma, multiple myeloma
(MM), amyloidosis, MPD, essential thrombocytosis (ET),
myelofibrosis (MF), polycythemia vera (PV), chronic myelomonocytic
leukemia (CMML), myelodysplastic syndrome (MDS), angioimmunoblastic
lymphoma, high-risk MDS, and low-risk MDS.
[0315] In one embodiment, the cancer is a T-cell lymphoma (e.g.,
peripheral T-cell lymphoma (PTCL) and cutaneous T-cell lymphoma
(CTCL)).
[0316] In one embodiment, the cancer is chronic lymphocytic
leukemia (CLL).
[0317] In other embodiments, the cancer has a high level of Bcl-2
expression. In one embodiment, the cancer is a large cell lymphoma
with a high level of Bcl-2 expression.
[0318] In one embodiment, the hematologic malignancy is relapsed.
In one embodiment, the hematologic malignancy is refractory.
[0319] In one embodiment, the cancer or disease is in a pediatric
patient (including an infantile patient). In one embodiment, the
cancer or disease is in an adult patient. Additional embodiments of
a cancer or disease being treated or prevented by methods,
compositions, or kits provided herein are described herein
elsewhere.
[0320] In exemplary embodiments, the cancer or hematologic
malignancy is CLL. In exemplary embodiments, the cancer or
hematologic malignancy is CLL/SLL. In exemplary embodiments, the
cancer or hematologic malignancy is blast phase CLL. In exemplary
embodiments, the cancer or hematologic malignancy is SLL.
[0321] In exemplary embodiments, the cancer or hematologic
malignancy is iNHL. In exemplary embodiments, the cancer or
hematologic malignancy is DLBCL. In exemplary embodiments, the
cancer or hematologic malignancy is B-cell NHL (e.g., aggressive
B-cell NHL). In exemplary embodiments, the cancer or hematologic
malignancy is MCL. In exemplary embodiments, the cancer or
hematologic malignancy is RS. In exemplary embodiments, the cancer
or hematologic malignancy is AML. In exemplary embodiments, the
cancer or hematologic malignancy is MM. In exemplary embodiments,
the cancer or hematologic malignancy is ALL. In exemplary
embodiments, the cancer or hematologic malignancy is T-ALL. In
exemplary embodiments, the cancer or hematologic malignancy is
B-ALL. In exemplary embodiments, the cancer or hematologic
malignancy is TCL. In exemplary embodiments, the cancer or
hematologic malignancy is ALCL. In exemplary embodiments, the
cancer or hematologic malignancy is leukemia. In exemplary
embodiments, the cancer or hematologic malignancy is lymphoma. In
exemplary embodiments, the cancer or hematologic malignancy is
T-cell lymphoma. In exemplary embodiments, the cancer or
hematologic malignancy is MDS (e.g., low grade MDS). In exemplary
embodiments, the cancer or hematologic malignancy is MPD. In
exemplary embodiments, the cancer or hematologic malignancy is a
mast cell disorder. In exemplary embodiments, the cancer or
hematologic malignancy is Hodgkin lymphoma (HL). In exemplary
embodiments, the cancer or hematologic malignancy is non-Hodgkin
lymphoma. In exemplary embodiments, the cancer or hematologic
malignancy is PTCL. In exemplary embodiments, the cancer or
hematologic malignancy is CTCL (e.g., mycosis fungoides or Sezary
syndrome). In exemplary embodiments, the cancer or hematologic
malignancy is WM. In exemplary embodiments, the cancer or
hematologic malignancy is CML. In exemplary embodiments, the cancer
or hematologic malignancy is FL. In exemplary embodiments, the
cancer or hematologic malignancy is transformed mycosis fungoides.
In exemplary embodiments, the cancer or hematologic malignancy is
Sezary syndrome. In exemplary embodiments, the cancer or
hematologic malignancy is acute T-cell leukemia. In exemplary
embodiments, the cancer or hematologic malignancy is acute B-cell
leukemia. In exemplary embodiments, the cancer or hematologic
malignancy is Burkitt lymphoma. In exemplary embodiments, the
cancer or hematologic malignancy is myeloproliferative neoplasms.
In exemplary embodiments, the cancer or hematologic malignancy is
splenic marginal zone. In exemplary embodiments, the cancer or
hematologic malignancy is nodal marginal zone. In exemplary
embodiments, the cancer or hematologic malignancy is extranodal
marginal zone.
[0322] In one embodiment, the cancer or hematologic malignancy is a
B cell lymphoma. In a specific embodiment, provided herein is a
method of treating or managing a B cell lymphoma comprising
administering to a patient a therapeutically effective amount of a
compound provided herein, or a pharmaceutically acceptable
derivative (e.g., salt or solvate) thereof. Also provided herein is
a method of treating or lessening one or more of the symptoms
associated with a B cell lymphoma comprising administering to a
patient a therapeutically effective amount of a compound provided
herein, or a pharmaceutically acceptable derivative (e.g., salt or
solvate) thereof. In one embodiment, the B cell lymphoma is iNHL.
In another embodiment, the B cell lymphoma is follicular lymphoma.
In another embodiment, the B cell lymphoma is Waldenstrom
macroglobulinemia (lymphoplasmacytic lymphoma). In another
embodiment, the B cell lymphoma is marginal zone lymphoma (MZL) In
another embodiment, the B cell lymphoma is MCL. In another
embodiment, the B cell lymphoma is HL. In another embodiment, the B
cell lymphoma is aNHL. In another embodiment, the B cell lymphoma
is DLBCL. In another embodiment, the B cell lymphoma is Richters
lymphoma.
[0323] In one embodiment, the cancer or hematologic malignancy is a
T cell lymphoma. In a specific embodiment, provided herein is a
method of treating (e.g., inhibiting, preventing or managing) a T
cell lymphoma comprising administering to a patient a composition
described herein. Also provided herein is a method of treating
(e.g., lessening one or more of the symptoms associated with) a T
cell lymphoma comprising administering to a patient a composition
described herein. In one embodiment, the T cell lymphoma is
peripheral T cell lymphoma (PTCL). In another embodiment, the T
cell lymphoma is cutaneous T cell lymphoma (CTCL).
[0324] In one embodiment, the cancer or hematologic malignancy is
Sezary syndrome. In a specific embodiment, provided herein is a
method of treating (e.g., inhibiting, preventing or managing)
Sezary syndrome comprising administering to a patient a composition
described herein. Also provided herein is a method of treating
(e.g., lessening one or more of the symptoms associated with)
Sezary syndrome comprising administering to a patient a
therapeutically effective amount of a composition described herein.
The symptoms associated with Sezary syndrome include, but are not
limited to, epidermotropism by neoplastic CD4+ lymphocytes,
Pautrier's microabscesses, erythroderma, lymphadenopathy, atypical
T cells in the peripheral blood, and hepatosplenomegaly. In one
embodiment of the compositions and methods described herein, the
dosage (e.g., the therapeutically effective dosage) of Compound 1
included in the combination is about 25 mg to 75 mg of Compound 1,
administered twice daily. In other embodiments, the dosage is from
about 50 mg to about 75 mg, from about 30 mg to about 65 mg, from
about 45 mg to about 60 mg, from about 30 mg to about 50 mg, or
from about 55 mg to about 65 mg. In certain embodiments, this
dosage is administered twice daily. In one embodiment, the dosage
is about 60 mg, administered twice daily.
Methods of Assessing Cancers
[0325] Accordingly, the present invention also provides a method of
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, comprising: assessing the level of a
prognosis-positive biomarker expressed by a tumor cell; and
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, wherein high expression levels of tumor cell
prognosis-positive biomarkers correlate with high sensitivity to
inhibition by a PI3K inhibitor, or wherein low expression levels of
said tumor cell prognosis-positive biomarker correlate with low
sensitivity to inhibition by PI3K inhibitors, and treating the
tumor with a PI3K inhibitor and a Bcl-2 inhibitor (e.g., if the
tumor is predicted to have a low sensitivity to a PI3K inhibitor
alone).
[0326] The present invention also provides a method of predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, comprising: assessing the level of a prognosis-negative
biomarker expressed by a tumor cell; and predicting the sensitivity
of tumor cell growth to inhibition by a PI3K inhibitor, wherein
high expression levels of tumor cell prognosis-negative biomarkers
correlate with low sensitivity to inhibition by PI3K inhibitors, or
wherein low expression levels of said tumor cell prognosis-negative
biomarker correlates with high sensitivity to inhibition by a PI3K
inhibitor, and treating the tumor with a PI3K inhibitor and a Bcl-2
inhibitor (e.g., if the tumor is predicted to have a low
sensitivity to a PI3K inhibitor alone).
[0327] The present invention further provides a method for treating
a tumor in a patient, comprising the step of administering to the
patient a PI3K inhibitor and a Bcl-2 inhibitor, wherein the patient
possesses a tumor that has been determined as having high
sensitivity to tumor cell growth inhibition by a PI3K inhibitor by
assessing the level of at least one prognosis-positive biomarker
expressed by a tumor cell from said tumor; and predicting the
sensitivity of tumor cell growth to inhibition by a PI3K inhibitor,
wherein high expression levels of said tumor cell
prognosis-positive biomarker correlate with high sensitivity to
inhibition by a PI3K inhibitor; or
[0328] assessing the level of at least one prognosis-negative
biomarker expressed by a tumor cell from said tumor; and predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, wherein low expression levels of said tumor cell
prognosis-negative biomarker correlate with high sensitivity to
inhibition by a PI3K inhibitor.
[0329] For any given prognosis-positive or prognosis-negative
biomarker, the range of expression level between tumor cells that
are relatively insensitive to PI3K inhibitors and those that are
sensitive, can readily be assessed by one of skill in the art, for
example by testing on a panel of tumor cells as described herein,
or by testing in tumor biopsies from patients whose tumors display
a range of sensitivities to a PI3K inhibitor.
[0330] One of skill in the medical arts, particularly pertaining to
the application of prognostic tests and treatment with
therapeutics, will recognize that biological systems are somewhat
variable and not always entirely predictable, and thus many good
diagnostic tests or therapeutics are occasionally ineffective.
Thus, it is ultimately up to the judgment of the attending
physician to determine the most appropriate course of treatment for
an individual patient, based upon test results, patient condition
and history, and his own experience. There may even be occasions,
for example, when a physician will choose to treat a patient with a
PI3K inhibitor even when a tumor is not predicted to be
particularly sensitive to PI3K inhibitors, based on data from
diagnostic tests or from other criteria, particularly if all or
most of the other obvious treatment options have failed, or if some
synergy is anticipated when given with another treatment. The fact
that the PI3K inhibitors as a class of compounds are relatively
well tolerated compared to many other anti-cancer compounds, such
as more traditional chemotherapy or cytotoxic agents used in the
treatment of cancer, makes this a more viable option.
[0331] Furthermore, this invention also provides additional methods
wherein simultaneous assessment of the expression level in tumor
cells of more than one biomarker level is utilized.
[0332] Accordingly, the present invention provides a method of
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, comprising: assessing the level of at least one (or
a panel of) prognosis-positive biomarkers expressed by a tumor
cell; and predicting the sensitivity of tumor cell growth to
inhibition by a PI3K inhibitor, wherein simultaneous high
expression levels of all of the assessed tumor cell
prognosis-positive biomarkers correlates with high sensitivity to
inhibition by a PI3K inhibitor, and treating the tumor with a PI3K
inhibitor and a Bcl-2 inhibitor (e.g., if the tumor is predicted to
have a low sensitivity to a PI3K inhibitor alone).
[0333] The present invention also provides a method of predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, comprising: assessing the level of one or more (or a
panel of) prognosis-negative biomarkers expressed by a tumor cell;
and predicting the sensitivity of tumor cell growth to inhibition
by a PI3K inhibitor, wherein simultaneous low or undetectable
expression levels of all of the assessed tumor cell
prognosis-negative biomarkers correlates with high sensitivity to
inhibition by a PI3K inhibitor, and treating the tumor with a PI3K
inhibitor and a Bcl-2 inhibitor (e.g., if the tumor is predicted to
have a low sensitivity to a PI3K inhibitor alone).
[0334] The present invention also provides a method of predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, comprising: assessing the level of one or more
prognosis-positive biomarker expressed by a tumor cell; assessing
the level of one or more prognosis-negative biomarker expressed by
a tumor cell; and predicting the sensitivity of tumor cell growth
to inhibition by a PI3K inhibitor, wherein a high ratio of
prognosis-positive to prognosis-negative biomarker expression
levels correlates with high sensitivity to inhibition by a PI3K
inhibitor and treating the tumor with a PI3K inhibitor and a Bcl-2
inhibitor (e.g., if the tumor is predicted to have a low
sensitivity to a PI3K inhibitor alone). As used herein, a high
ratio of prognosis-positive to prognosis-negative biomarker
expression levels means greater than 1:1, preferably greater than
1.1:1, preferably greater than 1.5:1, more preferably greater than
2:1, more preferably greater than 5:1, more preferably greater than
10:1, even more preferably greater than 100:1, or greater than
1,000:1.
[0335] In methods of this invention, biomarker expression level can
be assessed relative to the biomarker level in non-tumor cells of
the same tissue, or another cell or tissue source used as an assay
reference. The expression level of a biomarker is considered high
if expression level relative to a suitable reference is greater
than 1:1, preferably greater than 1.1:1, preferably greater than
1.5:1, more preferably greater than 2:1, more preferably greater
than 5:1, more preferably greater than 10:1, even more preferably
greater than 100:1, even more preferably greater than 1,000:1, even
more preferably greater than 10,000:1, even more preferably greater
than 1,000,000:1. The expression level of a biomarker is considered
low if expression level relative to a suitable reference is less
than 1:1, preferably less than 1:1.1, preferably less than 1:1.5,
more preferably less than 1:2, more preferably less than 1:5, more
preferably less than 1:10, even more preferably less than 1:100,
even more preferably less than 1:1,000, even more preferably less
than 1:10,000, even more preferably less than 1:1,000,000.
[0336] The present invention further provides a method of
predicting the likelihood that a tumor will progress to a more
aggressive tumor wherein the tumor is treatable with a PI3K
inhibitor, comprising: assessing the level of at least one
progression-positive biomarker expressed by a tumor cell from said
tumor; and predicting the likelihood that the tumor cell will
progress to a more aggressive tumor, wherein high expression levels
of said tumor cell progression-positive biomarker correlate with
high likelihood that the tumor cell will progress to a more
aggressive tumor or wherein low expression levels of said tumor
cell progression-positive biomarker correlate with low likelihood
that the tumor cell will progress to a more aggressive tumor and
treating the tumor with a PI3K inhibitor and a Bcl-2 inhibitor
(e.g., if it is determined that there is a high likelihood that the
tumor cell will progress to a more aggressive tumor if treated with
a PI3K inhibitor alone).
[0337] In one embodiment, the PI3K inhibitor is selected from
Compound 1, GS1101, BKM 120, GDC-0941, PX-866, GDC-0032, BAY
80-6946, BEZ235, BYL719, BGT-226, PF-4691502, GDC-0980, GSK
2126458, PF-05212384, XL765, or XL147. In some embodiments, the
PI3K inhibitor is selected from Compound 1 and GS1101. In some
embodiments, the PI3K inhibitor is Compound 1. In one embodiment
the tumor or tumor cell is selected from chronic lymphocytic
leukemia, non-Hodgkin lymphoma, diffuse large B-cell lymphoma,
mantle cell lymphoma, and adult T-cell lymphoma. In certain
embodiments, the tumor is selected from chronic lymphocytic
leukemia, non-Hodgkin lymphoma and diffuse large B-cell lymphoma.
In one embodiment, the PI3K inhibitor is Compound 1 and the tumor
or tumor cell is indolent non-Hodgkin lymphoma. In one embodiment
the progression-positive biomarker is a genomic alteration in one
or more gene in the 6q deletion region. In one embodiment, the
progression-positive biomarker is a genomic alteration in an
NF-.kappa.B pathway gene. In one embodiment, the
progression-positive biomarker is a del(6q13-16) or a del(6q23-24).
In one embodiment the progression-positive biomarker is a TNFAIP3
mutation or copy number loss. In one embodiment the
progression-positive biomarker is an EPHA7 mutation or copy number
loss.
[0338] The present invention also provides a method of predicting
the likelihood that a tumor cell from a tumor will progress to a
more aggressive tumor wherein the tumor is treatable with a PI3K
inhibitor, comprising: assessing the level of at least one
progression-negative biomarker expressed by a tumor cell; and
predicting the likelihood that the tumor cell will progress to a
more aggressive tumor, wherein high expression levels of said tumor
cell progression-negative biomarker correlate with low likelihood
that the tumor cell will progress to a more aggressive tumor, or
wherein low expression levels of said tumor cell
progression-negative biomarker correlates with high sensitivity to
inhibition by a PI3K inhibitor and treating the tumor with a PI3K
inhibitor and a Bcl-2 inhibitor (e.g., if it is determined that
there is a high likelihood that the tumor cell will progress to a
more aggressive tumor if treated with a PI3K inhibitor alone). In
one embodiment, the PI3K inhibitor is selected from Compound 1,
GS1101, BKM 120, GDC-0941, PX-866, GDC-0032, BAY 80-6946, BEZ235,
BYL719, BGT-226, PF-4691502, GDC-0980, GSK 2126458, PF-05212384,
XL765, or XL147. In some embodiments, the PI3K inhibitor is
selected from Compound 1 and GS1101. In certain embodiments, the
PI3K inhibitor is Compound 1. In one embodiment the tumor or tumor
cell is selected from chronic lymphocytic leukemia, non-Hodgkin
lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, and
adult T-cell lymphoma. In some embodiments, the tumor is selected
from chronic lymphocytic leukemia, non-Hodgkin lymphoma and diffuse
large B-cell lymphoma. In one embodiment, the PI3K inhibitor is
Compound 1 and the tumor or tumor cell is indolent non-Hodgkin
lymphoma. In one embodiment the progression-positive biomarker is a
genomic alteration in one or more gene in the 6q deletion region.
In one embodiment, the progression-positive biomarker is a genomic
alteration in an NF-.kappa.B pathway gene. In one embodiment, the
progression-positive biomarker is a del(6q13-16) or a del(6q23-24).
In one embodiment the progression-positive biomarker is a TNFAIP3
mutation or copy number loss. In one embodiment the
progression-positive biomarker is an EPHA7 mutation or copy number
loss.
[0339] In a further aspect, the present invention provides a method
for treating a tumor in a patient, comprising the step of
administering to the patient a PI3K inhibitor and a Bcl-2
inhibitor, wherein there is a high likelihood that the patient will
develop a more aggressive tumor and wherein said likelihood has
been determined by:
[0340] assessing the level of at least one progression-positive
biomarker expressed by a tumor cell from said tumor; and predicting
the likelihood that the tumor cell will progress to a more
aggressive tumor, wherein high expression levels of said tumor cell
progression-positive biomarker correlate with high likelihood that
the tumor cell will progress to a more aggressive tumor; or
[0341] assessing the level of at least one progression-negative
biomarker expressed by a tumor cell from said tumor; and predicting
the likelihood that the tumor cell will progress to a more
aggressive tumor, wherein low expression levels of said tumor cell
progression-negative biomarker correlate with high likelihood that
the tumor cell will progress to a more aggressive tumor.
[0342] In one embodiment, the PI3K inhibitor is selected from
Compound 1, GS1101, BKM 120, GDC-0941, PX-866, GDC-0032, BAY
80-6946, BEZ235, BYL719, BGT-226, PF-4691502, GDC-0980, GSK
2126458, PF-05212384, XL765, or XL147. In some embodiments, the
PI3K inhibitor is selected from Compound 1 and GS1101. In certain
embodiments, the PI3K inhibitor is Compound 1. In one embodiment
the tumor or tumor cell is selected from chronic lymphocytic
leukemia, non-Hodgkin lymphoma, diffuse large B-cell lymphoma,
mantle cell lymphoma, and adult T-cell lymphoma. In some
embodiments, the tumor is selected from chronic lymphocytic
leukemia, non-Hodgkin lymphoma and diffuse large B-cell lymphoma.
In one embodiment, the PI3K inhibitor is Compound 1 and the tumor
or tumor cell is indolent non-Hodgkin lymphoma. In one embodiment
the progression-positive biomarker is a genomic alteration in one
or more gene in the 6q deletion region. In one embodiment, the
progression-positive biomarker is a genomic alteration in an
NF-.kappa.B pathway gene. In one embodiment, the
progression-positive biomarker is a del(6q13-16) or a del(6q23-24).
In one embodiment the progression-positive biomarker is a TNFAIP3
mutation or copy number loss. In one embodiment the
progression-positive biomarker is an EPHA7 mutation or copy number
loss.
[0343] In the methods of this invention, the level of a
prognosis-positive or prognosis-negative biomarker expressed by a
tumor cell can be assessed by using any of the standard bioassay
procedures known in the art for determination of the level of
expression of a gene, including for example ELISA, RIA,
immunoprecipitation, immunoblotting, immunofluorescence microscopy,
RT-PCR, in situ hybridization, cDNA microarray, or the like, as
described in more detail below.
[0344] In the methods of this invention, the expression level of a
tumor cell prognosis-positive biomarker or prognosis-negative
biomarker is preferably assessed by assaying a tumor biopsy.
However, in an alternative embodiment, expression level of the
tumor cell biomarker can be assessed in bodily fluids or excretions
containing detectable levels of biomarkers originating from the
tumor or tumor cells. Bodily fluids or excretions useful in the
present invention include blood, urine, saliva, stool, pleural
fluid, lymphatic fluid, sputum, ascites, prostatic fluid,
cerebrospinal fluid (CSF), or any other bodily secretion or
derivative thereof. By blood it is meant to include whole blood,
plasma, serum or any derivative of blood. Assessment of tumor
prognosis-positive or prognosis-negative biomarkers in such bodily
fluids or excretions can sometimes be preferred in circumstances
where an invasive sampling method is inappropriate or
inconvenient.
[0345] In any of the above methods referring to a patient sample,
an example of such a sample can be a tumor biopsy.
[0346] In one embodiment, the biomarkers provided herein include,
but are not limited to, a target biomarker, a signaling pathway
biomarker, a protein mutation biomarker, a protein expression
biomarker, a gene mutation biomarker, a copy number alteration
(CNA) biomarker, a gene expression biomarker, a cytokine biomarker,
a chemokine biomarker, a matrix metalloproteinase biomarker, or a
biomarker for particular cancer cells. In one embodiment, the
biomarker can be used to evaluate the prognosis, and/or sensitivity
to a treatment agent, of a particular type of cancer or disease, or
of a particular patient or group of patients.
[0347] In one embodiment, the prognosis-positive or
prognosis-negative biomarker is a genomic alteration. In one
embodiment, the genomic alteration is a gene mutation or a copy
number alteration. In one embodiment, the gene mutation is a
non-dbSNP mutation. In another embodiment, the gene mutation is a
single nucleotide polymorphism (SNP) mutation. In one embodiment,
the prognosis-negative biomarker is associated with a mutation in
one or more of the following genes: ALK, SF3B1, TP53, NOTCH1,
MYD88, ATM, XPO1, POT1, NRAS, BCOR, KRAS, MED12, DDX3X, FBXW7, BTK
and PLCG2. In one embodiment, the prognosis-negative biomarker is
associated with a mutation in one or more of the following genes:
SF3B1, TP53, NOTCH1, MYD88, ATM, XPO1, MED12, and FBXW7. In one
embodiment, the prognosis-negative biomarker is associated with a
chromosome deletion.
[0348] In one embodiment, the prognosis-negative biomarker is
associated with one or more genomic alterations selected from the
group consisting of del(11q21), del(13q14), trisomy 12,
del(11q22-23), del(17p13), del(8p), TP53 mutation, TP53 pathway
mutation, MAPK pathway mutation, TP53 copy number loss, STK11 copy
number loss, TSC1 copy number loss, and TSC2 copy number loss. In
one embodiment, the prognosis-negative biomarker is copy number
loss in one or more of STK11, TSC1, and TSC2. In one embodiment,
the prognosis-negative biomarker is copy number loss in STK11. In
one embodiment, the prognosis-negative biomarker is copy number
loss in TSC1. In one embodiment, the prognosis-negative biomarker
is copy number loss in TSC2. In one embodiment, the
prognosis-negative biomarker is copy number loss in STK11 and TSC1.
In one embodiment, the prognosis-negative biomarker is copy number
loss in STK11 and TSC2. In one embodiment, the prognosis-negative
biomarker is TP53 pathway mutation or MAPK pathway mutation or
both. In one embodiment, the prognosis-negative biomarker is TP53
pathway and MAPK pathway dual mutation. In one embodiment, the
prognosis-negative biomarker is TP53 C141Y mutation. In another
embodiment, the prognosis-negative biomarker is ALK E1028D
mutation.
[0349] In one embodiment, the prognosis-negative biomarker is
associated with one or more (e.g., 2, 3, 4, 5, or all) genomic
alterations selected from the group consisting of del(11q21),
del(13q14), trisomy 12, del(11q22-23), del(17p13), and del(8p).
[0350] In an embodiment, the prognosis-negative biomarker is one or
more genomic alterations selected from the group consisting of BRAF
copy number gain, CTNNB1 copy number gain, FHIT copy number gain,
IRF4 copy number gain, MITF copy number gain, MN1 copy number gain,
NF2 copy number gain, NF2 copy number loss, RET copy number loss,
STK11 copy number loss, TSC2 copy number loss, RB1 loss of
heterozygosity.
[0351] In an embodiment, the prognosis-positive biomarker is one or
more of RANBP17 copy number gain, FGFR3 loss of heterozygosity,
GMPS loss of heterozygosity, and WHSC1 loss of heterozygosity.
[0352] In one embodiment, the progression-positive or
progression-negative biomarker is a genomic alteration. In one
embodiment, the genomic alteration is a gene mutation or a copy
number alteration. In one embodiment, the gene mutation is a
non-dbSNP mutation. In another embodiment, the gene mutation is a
single nucleotide polymorphism (SNP) mutation. In one embodiment,
the progression-positive biomarker is a genomic alteration in one
or more gene in the 6q deletion region. In an embodiment of the
invention the progression-positive biomarker is a genomic
alteration in an NF-.kappa.B pathway gene. In an embodiment, the
progression-positive biomarker is a del(6q13-16) or a del(6q23-24).
In one embodiment the progression-positive biomarker is a TNFAIP3
mutation or copy number loss. In one embodiment the
progression-positive biomarker is an EPHA7 mutation or copy number
loss.
[0353] In certain aspects provided herein is a method of predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, comprising: assessing the level of at least one
prognosis-positive biomarker expressed by a tumor cell; and
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, wherein high levels of a prognosis-positive
biomarker expression by the tumor cells correlates with high
sensitivity to inhibition by a PI3K inhibitor, or wherein low
expression levels of said tumor cell prognosis-positive biomarker
correlate with low sensitivity to inhibition by PI3K inhibitors,
and treating the tumor with a PI3K inhibitor and a Bcl-2
inhibitor.
[0354] In certain aspects, provided herein is a method of
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, comprising: assessing the level of at least one
prognosis-negative biomarker expressed by a tumor cell; and
predicting the sensitivity of tumor cell growth to inhibition by a
PI3K inhibitor, wherein high levels of prognosis-negative biomarker
expression by the tumor cell correlates with low sensitivity to
inhibition by a PI3K inhibitor, or wherein low expression levels of
said tumor cell prognosis-negative biomarker correlates with high
sensitivity to inhibition by a PI3K inhibitor, and treating the
tumor with a PI3K inhibitor and a Bcl-2 inhibitor.
[0355] In certain aspects provided herein is a method for treating
a tumor in a patient comprising the step of administering to the
patient a PI3K inhibitor, wherein the patient possesses a tumor
that has been determined as having high sensitivity to tumor cell
growth inhibition by a PI3K inhibitor by (a) assessing the level of
at least one prognosis-positive biomarker expressed by a tumor cell
from said tumor; and predicting the sensitivity of tumor cell
growth to inhibition by a PI3K inhibitor, wherein high expression
levels of said tumor cell prognosis-positive biomarker correlate
with high sensitivity to inhibition by a PI3K inhibitor; or (b)
assessing the level of at least one prognosis-negative biomarker
expressed by a tumor cell from said tumor; and predicting the
sensitivity of tumor cell growth to inhibition by a PI3K inhibitor,
wherein low expression levels of said tumor cell prognosis-negative
biomarker correlate with high sensitivity to inhibition by a PI3K
inhibitor, and treating the tumor with a PI3K inhibitor and a Bcl-2
inhibitor.
[0356] In certain aspects, provided herein is a method for treating
a tumor in a patient comprising the step of administering to the
patient a PI3K inhibitor as a first-line therapy, wherein the
patient possesses a tumor that has been determined as having high
sensitivity to tumor cell growth inhibition by a PI3K inhibitor by
(a) assessing the level of at least one prognosis-positive
biomarker expressed by a tumor cell from said tumor; and predicting
the sensitivity of tumor cell growth to inhibition by a PI3K
inhibitor, wherein high expression levels of said tumor cell
prognosis-positive biomarker correlate with high sensitivity to
inhibition by a PI3K inhibitor; or (b) assessing the level of at
least one prognosis-negative biomarker expressed by a tumor cell
from said tumor; and predicting the sensitivity of tumor cell
growth to inhibition by a PI3K inhibitor, wherein low expression
levels of said tumor cell prognosis-negative biomarker correlate
with high sensitivity to inhibition by a PI3K inhibitor, and
treating the tumor with a PI3K inhibitor and a Bcl-2 inhibitor.
[0357] In some embodiments, the PI3K inhibitor can be selected from
Compound 1, GS1101, BKM 120, GDC-0941, PX-866, GDC-0032, BAY
80-6946, BEZ235, BYL719, BGT-226, PF-4691502, GDC-0980, GSK
2126458, PF-05212384, XL765, or XL147.
[0358] In some embodiments, the PI3K inhibitor is selected from
Compound 1 and GS1101.
[0359] In some embodiments, the tumor is an acoustic neuroma,
adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma,
benign monoclonal gammopathy, biliary cancer bladder cancer, breast
cancer, brain cancer, bronchus cancer, cervical cancer,
choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer,
epithelial carcinoma, ependymoma, endotheliosarcoma, endometrial
cancer, esophageal cancer, Ewing sarcoma, familiar
hypereosinophilia, gastric cancer, gastrointestinal stromal tumor
(GIST), head and neck cancer, oral cancer, heavy chain disease,
hemangioblastoma, inflammatory myofibroblastic tumors, immunocytic
amyloidosis, kidney cancer, liver cancer, malignant hepatoma, lung
cancer, leiomyosarcoma (LMS), mastocytosis, multiple myeloma (MM),
myelodysplastic syndrome (MDS), mesothelioma, neuroblastoma,
neurofibroma neuroendocrine cancer, osteosarcoma, ovarian cancer,
Paget's disease of the vulva, Paget's disease of the penis,
papillary adenocarcinoma, pancreatic cancer, pinealoma, primitive
neuroectodermal tumor (PNT), prostate cancer, rhabdomyosarcoma,
retinoblastoma, salivary gland cancer, skin cancer, small bowel
cancer, soft tissue sarcoma, sebaceous gland carcinoma, sweat gland
carcinoma, synovioma, testicular cancer, thyroid cancer, and
Waldenstrom's macroglobulinemia.
[0360] In some embodiments, the tumor is a myeloid disorder,
lymphoid disorder, leukemia, lymphoma, myelodysplastic syndrome
(MDS), myeloproliferative disease (MPD), mast cell disorder, or a
myeloma.
[0361] In some embodiments, the tumor is selected from acute
lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, B-cell
acute lymphoblastic leukemia, acute myeloid leukemia, chronic
lymphocytic leukemia (CLL), chronic myelogenous leukemia, blast
phase chronic myelogenous leukemia, small lymphocytic lymphoma
(SLL), CLL/SLL, blast phase CLL, Hodgkin lymphoma, non-Hodgkin
lymphoma (NHL), B-cell NHL, T-cell NHL, indolent NHL, diffuse large
B-cell lymphoma, mantle cell lymphoma, aggressive B-cell NHL,
B-cell lymphoma, Richter's syndrome, T-cell lymphoma, peripheral
T-cell lymphoma, cutaneous T-cell lymphoma, transformed mycosis
fungoides, Sezary syndrome, anaplastic large-cell lymphoma,
follicular lymphoma, Waldenstrom macroglobulinemia,
lymphoplasmacytic lymphoma, Burkitt lymphoma, multiple myeloma,
amyloidosis, MPD, essential thrombocytosis, myelofibrosis,
polycythemia vera, chronic myelomonocytic leukemia, myelodysplastic
syndrome, angioimmunoblastic lymphoma, high-risk MDS, and low-risk
MDS.
[0362] In some embodiments, the tumor is selected from chronic
lymphocytic leukemia, non-Hodgkin lymphoma (e.g., indolent
Non-Hodgkin lymphoma), diffuse large B-cell lymphoma, mantle cell
lymphoma, and adult T-cell lymphoma.
[0363] In some embodiments, the prognosis-positive or
prognosis-negative biomarker is a genomic alteration.
[0364] In some embodiments, the prognosis-positive or
prognosis-negative biomarker is selected from a gene mutation, a
copy number alteration, a non-dbSNP mutation or an single
nucleotide polymorphism (SNP) mutation.
[0365] In some embodiments, the prognosis-positive biomarker is
associated with a mutation in a gene selected from RANBP17 copy
number gain, FGFR3 loss of heterozygosity, GMPS loss of
heterozygosity and WHSC1 loss of heterozygosity.
[0366] In some embodiments, the prognosis-negative biomarker is
associated with a genomic alteration selected from the group
consisting of del(11q21), del(13q14), del(8p), trisomy 12,
del(11q22-23), del(17p13), TP53 mutation, TP53 pathway mutation,
MAPK pathway mutation, TP53 copy number loss, STK11 copy number
loss, TSC1 copy number loss, and TSC2 copy number loss.
[0367] In some embodiments, the prognosis-negative biomarker is
associated with a mutation in a gene selected from SF3B1, TP53,
NOTCH1, MYD88, ATM, XPO1, POT1, NRAS, BCOR, KRAS, MED12, DDX3X,
FBXW7, BTK and PLCG2.
[0368] In some embodiments, the prognosis-negative biomarker is
associated with an STK11 copy number loss, a TSC1 or a TSC2 copy
number loss.
[0369] In certain aspects, provided herein is a method of
predicting the likelihood that a tumor will progress to a more
aggressive tumor wherein the tumor is treatable with a PI3K
inhibitor and a Bcl-2 inhibitor, said method comprising the steps
of: assessing the level of at least one progression-positive
biomarker expressed by a tumor cell from said tumor; and predicting
the likelihood that the tumor cell will progress to a more
aggressive tumor, wherein high expression levels of said tumor cell
progression-positive biomarker correlate with high likelihood that
the tumor cell will progress to a more aggressive tumor or wherein
low expression levels of said tumor cell progression-positive
biomarker correlate with low likelihood that the tumor cell will
progress to a more aggressive tumor.
[0370] In certain aspects, provided herein is a method of
predicting the likelihood that a tumor will progress to a more
aggressive tumor wherein the tumor is treatable with a PI3K
inhibitor and a Bcl-2 inhibitor, said method comprising the steps
of: assessing the level of at least one progression-negative
biomarker expressed by a tumor cell from said tumor; and predicting
the likelihood that the tumor cell will progress to a more
aggressive tumor, wherein high expression levels of said tumor cell
progression-negative biomarker correlate with low likelihood that
the tumor cell will progress to a more aggressive tumor or wherein
low expression levels of said tumor cell progression-positive
biomarker correlate with low likelihood that the tumor cell will
progress to a more aggressive tumor.
[0371] In certain aspects, provided herein is a method of treating
a tumor in a patient, comprising the step of administering to the
patient a PI3K inhibitor and a Bcl-2 inhibitor, wherein there is a
high likelihood that the patient will develop a more aggressive
tumor and wherein said likelihood has been determined by: (a)
assessing the level of at least one progression-positive biomarker
expressed by a tumor cell from said tumor; and predicting the
likelihood that the tumor cell will progress to a more aggressive
tumor, wherein high expression levels of said tumor cell
progression-positive biomarker correlate with high likelihood that
the tumor cell will progress to a more aggressive tumor; or (b)
assessing the level of at least one progression-negative biomarker
expressed by a tumor cell from said tumor; and predicting the
likelihood that the tumor cell will progress to a more aggressive
tumor, wherein low expression levels of said tumor cell
progression-negative biomarker correlate with high likelihood that
the tumor cell will progress to a more aggressive tumor.
[0372] In some embodiments, the PI3K inhibitor is selected from
Compound 1, GS1101, BKM 120, GDC-0941, PX-866, GDC-0032, BAY
80-6946, BEZ235, BYL719, BGT-226, PF-4691502, GDC-0980, GSK
2126458, PF-05212384, XL765, or XL147.
[0373] In any of the aforesaid aspects and embodiments, the tumor
is an acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal
cancer, angiosarcoma, benign monoclonal gammopathy, biliary cancer
bladder cancer, breast cancer, brain cancer, bronchus cancer,
cervical cancer, choriocarcinoma, chordoma, craniopharyngioma,
colorectal cancer, epithelial carcinoma, ependymoma,
endotheliosarcoma, endometrial cancer, esophageal cancer, Ewing
sarcoma, familiar hypereosinophilia, gastric cancer,
gastrointestinal stromal tumor (GIST), head and neck cancer, oral
cancer, heavy chain disease, hemangioblastoma, inflammatory
myofibroblastic tumors, immunocytic amyloidosis, kidney cancer,
liver cancer, malignant hepatoma, lung cancer, leiomyosarcoma
(LMS), mastocytosis, multiple myeloma (MM), myelodysplastic
syndrome (MDS), mesothelioma, neuroblastoma, neurofibroma
neuroendocrine cancer, osteosarcoma, ovarian cancer, Paget's
disease of the vulva, Paget's disease of the penis, papillary
adenocarcinoma, pancreatic cancer, pinealoma, primitive
neuroectodermal tumor (PNT), prostate cancer, rhabdomyosarcoma,
retinoblastoma, salivary gland cancer, skin cancer, small bowel
cancer, soft tissue sarcoma, sebaceous gland carcinoma, sweat gland
carcinoma, synovioma, testicular cancer, thyroid cancer, and
Waldenstrom's macroglobulinemia. In some embodiments, the tumor is
a myeloid disorder, lymphoid disorder, leukemia, lymphoma,
myelodysplastic syndrome (MDS), myeloproliferative disease (MPD),
mast cell disorder, or a myeloma. In some embodiments, the tumor is
indolent. In some embodiments, the tumor is selected from acute
lymphoblastic leukemia, T-cell acute lymphoblastic leukemia, B-cell
acute lymphoblastic leukemia, acute myeloid leukemia, chronic
lymphocytic leukemia (CLL), chronic myelogenous leukemia, blast
phase chronic myelogenous leukemia, small lymphocytic lymphoma
(SLL), CLL/SLL, blast phase CLL, Hodgkin lymphoma, non-Hodgkin
lymphoma (NHL), B-cell NHL, T-cell NHL, indolent NHL, diffuse large
B-cell lymphoma, mantle cell lymphoma, aggressive B-cell NHL,
B-cell lymphoma, Richter's syndrome, T-cell lymphoma, peripheral
T-cell lymphoma, cutaneous T-cell lymphoma, transformed mycosis
fungoides, Sezary syndrome, anaplastic large-cell lymphoma,
follicular lymphoma, Waldenstrom macroglobulinemia,
lymphoplasmacytic lymphoma, Burkitt lymphoma, multiple myeloma,
amyloidosis, MPD, essential thrombocytosis, myelofibrosis,
polycythemia vera, chronic myelomonocytic leukemia, myelodysplastic
syndrome, angioimmunoblastic lymphoma, high-risk MDS, and low-risk
MDS.
[0374] In some embodiments, the tumor is selected from chronic
lymphocytic leukemia, non-Hodgkin lymphoma (e.g., indolent
non-Hodgkin lymphoma), diffuse large B-cell lymphoma, mantle cell
lymphoma, and adult T-cell lymphoma.
[0375] In some embodiments, the progression-positive or
progression-negative biomarker is a genomic alteration.
[0376] In some embodiments, the progression-positive or
progression-negative biomarker is selected from a gene mutation, a
copy number alteration, a non-dbSNP mutation or an single
nucleotide polymorphism (SNP) mutation.
[0377] In some embodiments, the progression-positive biomarker is
associated with a mutation in a gene in the 6q deletion region.
[0378] In some embodiments, the progression-positive biomarker is a
genomic alteration in an NF-.kappa.B pathway gene.
[0379] In some embodiments, the progression-positive biomarker is a
del(6q13-16) or a del(6q23-24).
[0380] In some embodiments, the progression-positive biomarker is a
TNFAIP3 mutation or copy number loss.
[0381] In some embodiments, the progression-positive biomarker is
an EPHA7 mutation or copy number loss.
[0382] In some aspects, the disclosure provides a method of
treating a patient, comprising (i) administering a first treatment
comprising a first PI3K inhibitor to the subject (ii) acquiring
information regarding an alteration in a biomarker by comparing an
assessment of the biomarker in a first sample taken from the
subject before the first treatment is administered with an
assessment of the biomarker in a second sample taken from the
subject after the first treatment is administered, wherein the
biomarker is selected from STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3,
YWHAE, PER1, GAS7, FSTL3, USP6, MAP2K4, or EGFR, and (iii)
continuing administration of the first treatment if the alteration
is absent, or administering a second treatment if the alteration is
present, wherein the second treatment includes administration of a
Bcl-2 inhibitor.
[0383] In some aspects, the present disclosure provides a method of
determining the further course of treatment for a subject who has
undergone a first treatment with a first PI3K inhibitor, the method
comprising: (i) acquiring information regarding the presence or
absence of an alteration in one or more of STK11, TSC1, TSC2, TP53,
PTEN, CBFA2T3, YWHAE, PER1, GAS7, FSTL3, USP6, MAP2K4, or EGFR in
one or more samples from the subject; and (ii) selecting the
subject for continuation of the first treatment with the first PI3K
inhibitor if the alteration is absent and selecting the subject for
a second treatment if the alteration is present, wherein the second
treatment includes administration of a Bcl-2 inhibitor.
[0384] In some aspects, the disclosure provides a method of
determining decreased responsiveness, or resistance, of a subject
to a first treatment comprising a first PI3K inhibitor, the method
comprising (i) acquiring information regarding the presence or
absence of an alteration in one or more of STK11, TSC1, TSC2, TP53,
PTEN, CBFA2T3, YWHAE, PER1, GAS7, FSTL3, USP6, MAP2K4, or EGFR in
one or more samples from the subject; and (ii) determining that the
subject shows decreased responsiveness or resistance to the first
treatment if the alteration is present, wherein the second
treatment includes administration of a Bcl-2 inhibitor.
[0385] In any of the above aspects or embodiments, the PI3K
inhibitor can be selected from: Compound 1, AMG-319, GSK 2126458,
GSK 1059615, GDC-0032, GDC-0980, GDC-0941, XL147, XL499, XL765, BKM
120 GS1101, CAL 263, SF1126, PX-866, BEZ235, CAL-120, BYL719,
RP6503, RP6530, TGR1202, INK1117, PX-886, BAY 80-6946, IC87114,
Palomid 529, ZSTK474, PWT33597, TG100-115, GNE-477, CUDC-907,
AEZS-136, BGT-226, PF-05212384, LY3023414, PI-103, LY294002,
INCB-040093, CAL-130 and wortmannin.
[0386] The present invention also provides, at least in part,
methods (e.g., diagnostic and prognostic methods) for evaluating,
e.g., predicting, the responsiveness to a treatment of a cancer
with a B-cell receptor (BCR) pathway inhibitor (e.g., a PI3K
inhibitor). In one embodiment, it is shown herein that STK11 copy
number loss (with or without copy number loss of TSC1, TSC2, or
both) is associated with, or is predictive of, decreased
responsiveness (e.g., acquired resistance) of a cancer (e.g.,
chronic lymphocytic leukemia (CLL)) to a PI3K inhibitor (e.g.,
Compound 1). In other embodiments, it has been discovered that an
alteration in the MAP kinase and p53 (MAPK/p53) pathway is
associated with, or is predictive of, decreased responsiveness
(e.g., acquired resistance) of a cancer (e.g., CLL) to a PI3K
inhibitor (e.g., Compound 1). Thus, compositions, methods, and kits
for evaluating responsiveness (e.g., acquisition of resistance) to,
or monitor, therapy involving PI3K inhibition (including
combination therapies); stratify patient populations; identify
subjects likely to benefit from such agents, predict a time course
of disease or a probability of a significant event in the disease
for such subjects, and/or more effectively monitor, treat or
prevent a cancer are disclosed. The methods further comprise
treating the subject with a PI3K inhibitor and a Bcl-2
inhibitor.
[0387] Aspects of the invention disclosed herein are based, at
least in part, on the following findings. Additional details are
described herein in the Examples.
[0388] In experiments described herein, it was found that STK11
copy number loss is associated with or predictive of
nonresponsiveness or resistance (e.g., acquired resistance) of a
cancer (e.g., a CLL) to a PI3K inhibitor (e.g., Compound 1).
Furthermore, in experiments described herein, it was found that a
dual alteration in the MAPK/P53 pathway is associated with or
predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor (e.g.,
Compound 1).
[0389] In accordance with certain analyses described in the
Examples, it was found that copy number loss of STK11 combined with
copy number loss of TSC1, TSC2, or both is associated with or
predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor (e.g.,
Compound 1).
[0390] Also, in certain analyses described in the Examples, the
following relationships were revealed. TSC2 copy number loss was
associated with or predictive of nonresponsiveness or resistance
(e.g., acquired resistance) of a cancer (e.g., a CLL) to a PI3K
inhibitor (e.g., Compound 1). Copy number gain in each of BRAF,
CTNNB1, FHIT, IRF4, MITF, MN1, and NF2 was associated with or
predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor (e.g.,
Compound 1). Copy number loss in each of NF2 and RET was associated
with or predictive of nonresponsiveness or resistance (e.g.,
acquired resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor
(e.g., Compound 1). Loss of heterozygosity in RB1 was associated
with or predictive of nonresponsiveness or resistance (e.g.,
acquired resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor
(e.g., Compound 1). Copy number gain in RANBP17 was associated with
responsiveness or lack of resistance (e.g., acquired resistance) of
a cancer (e.g., a CLL) to a PI3K inhibitor (e.g., Compound 1). Loss
of heterozygosity in each of FGFR3, GMPS, and WHSC1 is associated
with or predictive of responsiveness or lack of resistance (e.g.,
acquired resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor
(e.g., Compound 1).
[0391] Improved methods for treating a cancer patient with a PI3K
inhibitor that incorporate the methods described herein are also
provided, whereby patients with high sensitivity to cancer or tumor
cell growth inhibition by a PI3K inhibitor are determined by the
methods of the present invention. Thus, the present invention
further provides a method for treating cancer in a subject, e.g., a
patient, comprising the step of administering to the subject a PI3K
inhibitor and a Bcl-2 inhibitor, wherein the subject possesses a
cancer that has been determined as having high sensitivity to
cancer or tumor cell growth inhibition by a PI3K inhibitor by
assessing the level of at least one prognosis-positive biomarker in
a cancer or tumor cell from said cancer or tumor; and predicting
the sensitivity of cancer or tumor cell growth to inhibition by a
PI3K inhibitor, wherein detection or an elevated level of said
prognosis-positive biomarker correlate with high sensitivity to
inhibition by a PI3K inhibitor; or assessing the level of at least
one prognosis-negative biomarker in a cancer or tumor cell from
said cancer or tumor; and predicting the sensitivity of cancer or
tumor cell growth to inhibition by a PI3K inhibitor, wherein the
presence or level of the alteration said prognosis-negative
biomarker correlate with high sensitivity to inhibition by a PI3K
inhibitor.
[0392] A further aspect of the invention is a method of treating a
cancer or tumor or a metastasis thereof in a subject, comprising
the step of administering to the subject a PI3K inhibitor, e.g., as
a first-line therapy, wherein the subject possesses a cancer or
tumor that has been determined as having high sensitivity to cancer
or tumor cell growth inhibition by a PI3K inhibitor by assessing
the level of at least one prognosis-positive biomarker by one of
the following: assessing the level of at least one
prognosis-positive biomarker expressed by a cancer cell from said
cancer or tumor; and predicting the sensitivity of cancer or tumor
cell growth to inhibition by a PI3K inhibitor, wherein detection or
an elevated level of said prognosis-positive biomarker correlate
with high sensitivity to inhibition by a PI3K inhibitor; or
assessing the presence or an alteration at least one
prognosis-negative biomarker in a cancer or tumor cell from said
cancer or tumor; and predicting the sensitivity of cancer or tumor
cell growth to inhibition by a PI3K inhibitor, wherein low levels
of said prognosis-negative biomarker correlate with high
sensitivity to inhibition by a PI3K inhibitor; and, if the cancer
or tumor cell growth is predicted to have low sensitivity to
inhibition by a PI3K inhibitor as a monotherapy, administering to
the patient a combination of a PI3K inhibitor and a Bcl-2
inhibitor.
[0393] In a further aspect, the present invention provides a method
for treating a cancer or tumor in a subject, e.g., a patient,
comprising administering to the subject a PI3K inhibitor and a
Bcl-2 inhibitor, wherein there is a high likelihood that the
patient will develop a more aggressive tumor and wherein said
likelihood has been determined by assessing the level of at least
one progression-positive biomarker expressed by a tumor cell from
said tumor; and predicting the likelihood that the tumor cell will
progress to a more aggressive tumor, wherein high expression levels
of said tumor cell progression-positive biomarker correlate with
high likelihood that the tumor cell will progress to a more
aggressive tumor; or assessing the level of at least one
progression-negative biomarker expressed by a tumor cell from said
tumor; and predicting the likelihood that the tumor cell will
progress to a more aggressive tumor, wherein low expression levels
of said tumor cell progression-negative biomarker correlate with
high likelihood that the tumor cell will progress to a more
aggressive tumor.
[0394] In certain aspects, the invention features a method of
evaluating the responsiveness of a cancer or tumor, or a subject
having a cancer or tumor, to a treatment with a BCR pathway
inhibitor (e.g., a treatment with an inhibitor of PI3K, BTK or SYK,
alone or in combination). In one embodiment, responsiveness to a
PI3K inhibitor is evaluated. The method includes: acquiring a value
(e.g., determining one or more of: the presence, absence, amount or
level) of an alteration or biomarker chosen from one, two, three,
four or all of: an STK11 copy number, TSC1 copy number, TSC2 copy
number, a p53 pathway mutation (e.g., a mutation disclosed in Table
20), or MAPK pathway mutation (e.g., a mutation disclosed in Table
18), or any combination thereof (e.g., a dual MAPK/p53 pathway
mutation, e.g., a mutation disclosed in Table 18 and a mutation
disclosed in Table 20). The method further comprises administering
a PI3K inhibitor and a Bcl-2 inhibitor to the subject.
[0395] In another aspect, the invention features a method of
monitoring a treatment of a subject with a BCR pathway inhibitor
(e.g., a treatment with an inhibitor of PI3K, BTK or SYK, alone or
in combination). In one embodiment, treatment with a PI3K inhibitor
is monitored. The method includes: acquiring, at two or more time
intervals, a value (e.g., determining one or more of: the presence,
absence, amount or level) of an alteration or biomarker chosen from
one, two, three, four or all of: an STK11 copy number, TSC1 copy
number, TSC2 copy number, a p53 pathway mutation (e.g., a mutation
disclosed in Table 20), or MAPK pathway mutation (e.g., a mutation
disclosed in Table 18), or any combination thereof (e.g., a dual
MAPK/p53 mutation, e.g., a mutation disclosed in Table 18 and a
mutation disclosed in Table 20). The method further comprises
administering a PI3K inhibitor and a Bcl-2 inhibitor to the
subject.
[0396] In another aspect, the invention features a method of
treating (e.g., inhibiting, reducing, ameliorating, managing, or
preventing) a cancer or tumor in a subject. The method includes:
acquiring a value (e.g., determining one or more of: the presence,
absence, amount or level) of an alteration or biomarker chosen from
one, two, three, four or all of: an STK11 copy number, TSC1 copy
number, TSC2 copy number, a p53 pathway mutation (e.g., a mutation
disclosed in Table 20), or MAPK pathway mutation (e.g., a mutation
disclosed in Table 18), or any combination thereof (e.g., a dual
MAPK/p53 mutation, e.g., a mutation disclosed in Table 18 and a
mutation disclosed in Table 20), and responsive to said value,
administering to the subject a BCR pathway inhibitor, e.g., a PI3K
inhibitor (e.g., one or more PI3K inhibitors) and a Bcl-2
inhibitor.
[0397] In another aspect, the invention features a method of
treating a subject, comprising (i) administering a first treatment
comprising a first PI3K inhibitor to the subject (ii) acquiring
information regarding the presence or absence of an alteration in a
biomarker in one or more samples from the subject, wherein the
biomarker is selected from STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3,
YWHAE, PER1, GAS7, FSTL3, USP6, MAP2K4, or EGFR; and (iii)
continuing administration of the first treatment if the alteration
is absent, or administering a second treatment if the alteration is
present, wherein the second treatment includes administration of a
Bcl-2 inhibitor.
[0398] In certain embodiments, the alteration is an STK11, TSC1,
TSC2, TP53, PTEN, CBFA2T3, YWHAE, PER1, GAS7, FSTL3, USP6, or
MAP2K4 copy number loss (e.g., single copy loss). In some
embodiments, the STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3, YWHAE,
PER1, GAS7, FSTL3, USP6, or MAP2K4 copy number in a sample taken
from the subject after the first treatment is lower than a
corresponding STK11, TSC1, TSC2, TP53, PTEN, CBFA2T3, YWHAE, PER1,
GAS7, FSTL3, USP6, MAP2K4 copy number in a sample taken from the
subject before the first treatment (e.g., there is an STK11 single
copy loss).
[0399] In another aspect, the present disclosure provides a method
of evaluating the responsiveness of a cancer or tumor, or a subject
having a cancer or tumor, to a treatment with a BCR pathway
inhibitor (e.g., a treatment with an inhibitor of PI3K, BTK or SYK,
alone or in combination). In one embodiment, responsiveness to a
PI3K inhibitor is evaluated. The method includes: acquiring a value
(e.g., determining one or more of: the presence, absence, amount or
level) of an anti-apoptotic factor such as Bcl-2.
[0400] In another aspect, the invention features a method of
monitoring a treatment of a subject with a BCR pathway inhibitor
(e.g., a treatment with an inhibitor of PI3K, BTK or SYK, alone or
in combination). In one embodiment, treatment with a PI3K inhibitor
is monitored. The method includes: acquiring, at two or more time
intervals, a value (e.g., determining one or more of: the presence,
absence, amount or level) of an anti-apoptotic factor such as
Bcl-2.
[0401] In another aspect, the invention features a method of
treating (e.g., inhibiting, reducing, ameliorating, managing, or
preventing) a cancer or tumor in a subject. The method includes:
acquiring a value (e.g., determining one or more of: the presence,
absence, amount or level) of an anti-apoptotic factor such as
Bcl-2.
[0402] In certain embodiments, the methods that include acquiring a
value of Bcl-2 also include acquiring a value (e.g., determining
one or more of: the presence, absence, amount or level) of a
pro-apoptotic factor or anti-apoptotic factor. The pro-apoptotic
factor can be, e.g., one or more of (e.g., 2, 3, 4, or all of) BMF,
BIK, BIM, NOXA, PUMA, and HRK. In some embodiments, an elevated
level of Bch 2 indicates that the cancer is resistant to a PI3K
inhibitor. In some embodiments, a normal or reduced level of Bcl-2
indicates that the cancer is responsive to a PI3K inhibitor. In
some embodiments, an elevated level of one or more pro-apoptotic
factors (e.g., BMF, BIK, BIM, NOXA, PUMA, and HRK) indicates that
the cancer is more responsive to a PI3K inhibitor (optionally in
combination with a Bcl-2 inhibitor) than a cancer with normal or
lowered levels of the pro-apoptotic factor. In some embodiments,
the methods involve administering a Bcl-2 inhibitor (e.g., in
combination with a PI3K inhibitor) to a subject having elevated
Bcl-2 levels. In some embodiments, the methods involve
administering a PI3K inhibitor as a monotherapy to a subject having
normal or low Bcl-2 levels. In some embodiments, the elevated,
normal, or reduced levels of a biomarker are determined with
reference to a non-cancerous control value.
[0403] In some embodiments, acquiring a value comprises acquiring
information regarding the presence or absence of an alteration
described herein.
[0404] In some embodiments, the methods herein comprise comparing
an assessment of a biomarker in a first sample taken from the
subject before the first treatment is administered with an
assessment of the biomarker in a second sample taken from the
subject after the first treatment is administered. In an
embodiment, the method comprises determining the further course of
treatment for the subject. In an embodiment, the method comprises a
method of determining decreased responsiveness, or resistance, of
the subject to the first treatment.
[0405] In some embodiments, the methods herein comprise
administering a first treatment comprising a first PI3K inhibitor
to the subject and continuing administration of the first treatment
if an alteration is absent, or administering a second treatment
including a Bcl-2 inhibitor, if the alteration is present. In some
embodiments, the methods herein comprise determining the further
course of treatment for a subject, e.g., selecting the subject for
continuation of the first treatment with the first PI3K inhibitor
if the alteration is absent and selecting the subject for a second
treatment if the alteration is present, wherein the second
treatment includes administration of a Bcl-2 inhibitor. In some
embodiments, the methods comprise determining decreased
responsiveness, or resistance, of a subject to a first treatment
comprising a first PI3K inhibitor. In some embodiments, the methods
comprise determining that the subject shows decreased
responsiveness or resistance to the first treatment if the
alteration is present, wherein the second treatment includes
administration of a Bcl-2 inhibitor.
[0406] In certain embodiments of the methods above, the alteration
is an STK11 copy number loss (e.g., single copy loss). In some
embodiments, the STK11 copy number in a sample taken from the
subject after the first treatment is lower than the STK11 copy
number in a sample taken from the subject before the first
treatment (e.g., there is an STK11 single copy loss). In some
embodiments, the first treatment with the first PI3K inhibitor
(e.g., Compound 1) is a monotherapy in which the first PI3K
inhibitor is the only component of the first treatment known to
have a substantial therapeutic activity.
[0407] In some embodiments of the methods herein, a
prognosis-negative biomarker is chosen from one, two, three or all
of the following:
[0408] (i) a copy number loss of STK11;
[0409] (ii) a copy number loss of TSC1 or TSC2, or both;
[0410] (iii) a p53 pathway mutation, e.g., TP53 C141Y; or
[0411] (iv) a MAPK pathway mutation.
[0412] In some embodiments of the methods herein, a
prognosis-negative biomarker is a copy number loss of STK11. In one
embodiment, detection of copy number loss of STK11 is indicative of
decreased responsiveness of the cancer or tumor, or the subject, to
the treatment.
[0413] In some embodiments of the methods herein, a
prognosis-negative biomarker is a dual MAPK/p53 mutation. In one
embodiment, detection of the dual MAPK/p53 mutation is indicative
of decreased responsiveness of the cancer or tumor, or the subject,
to the treatment.
[0414] In some embodiments of the methods herein, a
prognosis-negative biomarker is a copy number loss of STK11 in
combination with a copy number loss of TSC1, TSC2, or both. In one
embodiment, detection of copy number loss of STK11 in combination
with a copy number loss of TSC1 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In another embodiment, detection of copy number loss of
STK11 in combination with a copy number loss of TSC2 is indicative
of decreased responsiveness of the cancer or tumor, or the subject,
to the treatment. In yet another embodiment, detection of copy
number loss of STK11 in combination with a copy number loss of TSC1
and TSC2 is indicative of decreased responsiveness of the cancer or
tumor, or the subject, to the treatment.
[0415] In some embodiments of the methods herein, the alteration is
a prognosis-negative biomarker or a progression-positive biomarker,
or both. In one embodiment, detection of a prognosis-negative
biomarker or a progression-positive biomarker, or both, is
indicative of decreased responsiveness of the cancer or tumor, or
the subject, to the treatment.
[0416] In some embodiments of any of the above aspects, the BCR
pathway mutation is a mutation disclosed in Table 19. In an
embodiment, the p53 pathway mutation is a mutation disclosed in
Table 20. In an embodiment, the MAPK pathway mutation is a mutation
disclosed in Table 18. In an embodiment, the combination thereof is
a dual MAPK/p53 mutation of which a mutation is disclosed in Table
18 and a mutation is disclosed in Table 20.
[0417] In some embodiments of any of the above aspects, one, two,
three, four, five, six, seven, eight, nine, 10, 11, 12, 13, or all
of the following is indicative of decreased responsiveness of the
cancer, or the subject, to the treatment:
[0418] (i) a copy number loss (e.g., a single copy loss) of
STK11;
[0419] (ii) a copy number loss of TSC1 or TSC2, or both;
[0420] (iii) a copy number loss of TP53;
[0421] (iv) a copy number loss of PTEN;
[0422] (v) a copy number loss of CBFAT2T3;
[0423] (vi) a copy number loss of YWHAE;
[0424] (vii) a copy number loss of PER1;
[0425] (viii) a copy number loss of GAS7;
[0426] (ix) a copy number loss of FSTL3;
[0427] (x) a copy number loss of USP6;
[0428] (xi) a copy number loss of MAP2K4;
[0429] (xii) a BCR pathway mutation;
[0430] (xiii) a p53 pathway mutation, e.g., a mutation listed in
Table 20 (e.g., TP53 C141Y); or
[0431] (xiv) a MAPK pathway mutation, e.g., a mutation listed in
Table 18.
[0432] In some embodiments of any of the methods herein, the
alteration or biomarker is a copy number loss (e.g., a single copy
loss) of STK11. In one embodiment, detection of copy number loss of
STK11 is indicative of decreased responsiveness of the cancer or
tumor, or the subject, to the treatment. In some embodiments of any
of the above aspects, the alteration or biomarker is a dual
MAPK/p53 pathway mutation. In one embodiment the dual mutation
includes a mutation listed in Table 18 and/or Table 20. In one
embodiment, detection of the dual MAPK/p53 pathway mutation is
indicative of decreased responsiveness of the cancer or tumor, or
the subject, to the treatment. In some embodiments of any of the
above aspects, the alteration or biomarker is a BCR pathway
mutation. In one embodiment, detection of a BCR pathway mutation is
indicative of decreased responsiveness of the cancer or tumor, or
the subject, to the treatment. In some embodiments of any of the
above aspects, detection of copy number loss of TP53 is indicative
of decreased responsiveness of the cancer or tumor, or the subject,
to the treatment. In some embodiments of any of the above aspects,
detection of copy number loss of PTEN is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of CBFAT2T3 is indicative of
decreased responsiveness of the cancer or tumor, or the subject, to
the treatment. In some embodiments of any of the above aspects,
detection of copy number loss of YWHAE is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of PER1 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of GAS7 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of FSTL3 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of USP6 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of MAP2K4 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In some embodiments of any of the above aspects,
detection of copy number loss of EGFR is indicative of increased
responsiveness of the cancer or tumor, or the subject, to the
treatment; or wherein detection of copy number gain of EGFR is
indicative of decreased responsiveness of the cancer or tumor, or
the subject, to the treatment, or both. In some embodiments of any
of the above aspects, detection of copy number loss of EGFR is
indicative of increased responsiveness of the cancer or tumor, or
the subject, to the treatment, and wherein increased responsiveness
is determined using nodal criteria.
[0433] In some embodiments of any of the above aspects, the
alteration or biomarker is a copy number loss of STK11 in
combination with a copy number loss of TSC1, TSC2, or both. In one
embodiment, detection of copy number loss of STK11 in combination
with a copy number loss of TSC1 is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment. In another embodiment, detection of copy number loss of
STK11 in combination with a copy number loss of TSC2 is indicative
of decreased responsiveness of the cancer or tumor, or the subject,
to the treatment. In yet another embodiment, detection of copy
number loss of STK11 in combination with a copy number loss of TSC1
and TSC2 is indicative of decreased responsiveness of the cancer or
tumor, or the subject, to the treatment.
[0434] In some embodiments of any of the above aspects, no
detectable copy number loss of STK11, TSC1, TSC2, TP53, PTEN,
CBFA2T3, YWHAE, PER1, GAS7, FSTL3, USP6, or MAP2K4, or no
detectable dual MAPK/p53 pathway mutation, or no detectable BCR
pathway mutation, is indicative of continued responsiveness to the
treatment.
[0435] In some embodiments of any of the above aspects, the
alteration is a prognosis-negative biomarker or a
progression-positive biomarker, or both. In one embodiment,
detection of a prognosis-negative biomarker or a
progression-positive biomarker, or both, is indicative of decreased
responsiveness of the cancer or tumor, or the subject, to the
treatment.
[0436] In some embodiments of any of the above aspects, no
detectable copy number loss of STK11, or no detectable dual
MAPK/p53 pathway mutation, is indicative of continued
responsiveness to the treatment.
[0437] In some embodiments of any of the above aspects, the subject
is evaluated prior to undergoing, while undergoing, or after
undergoing, treatment with the BCR pathway inhibitor, e.g., the
PI3K inhibitor. In another embodiment, the subject is evaluated at
at least two time intervals, e.g., prior to undergoing and while
undergoing the treatment. In yet another embodiment, the subject is
evaluated at at least three time points, e.g., prior to undergoing,
while undergoing the treatment, and after undergoing the
treatment.
[0438] In some embodiments of any of the above aspects, decreased
responsiveness of the cancer or tumor, or the subject to the
treatment, e.g., over a timecourse of the treatment, is indicative
of increased resistance (e.g., acquired resistance) to the
treatment, e.g., the PI3K inhibitor. In an embodiment, if the
subject is identified as being responsive to the treatment, the
treatment is continued. In an embodiment, if the subject is
identified as not being responsive to the treatment, the treatment
is altered or discontinued, thereby having a first and second
treatment.
[0439] In some embodiments of the aforesaid methods, responsive to
a determination of the value of the alteration or biomarker, the
method further includes one, two, three, four, five, six, seven,
eight, nine or all of the following:
[0440] (i) identifying the subject as being in need of a treatment,
e.g., treatment with a PI3K inhibitor (e.g., a first treatment or a
second (alternative) treatment);
[0441] (ii) identifying the subject as having an increased or a
decreased responsiveness to the treatment, e.g., the treatment with
the PI3K inhibitor (e.g., a monotherapy with Compound 1);
[0442] (iii) identifying the subject as being a responder to the
treatment, e.g., identifying the subjects as being in complete
remission (CR) or partial cancer remission (PR) (e.g., CR or PR
subjects as described herein);
[0443] (iv) identifying the subject as being a non-responder to the
treatment, e.g., identifying the subjects as having a progressive
disease (PD) or stable disease (SD) (e.g., PD or SD subjects as
described herein);
[0444] (v) identifying the subject as having developed resistance
(e.g., partial or complete, acquired resistance) to the treatment,
e.g., the PI3K inhibitor (e.g., Compound 1);
[0445] (vi) diagnosing and/or prognosing the subject;
[0446] (vii) determining a time course of disease progression in
the subject;
[0447] (viii) determining the time course of acquisition of
resistance to the treatment;
[0448] (ix) determining a treatment, e.g., selecting or altering
the course of, a treatment (e.g., a first treatment), a dose, a
treatment schedule or time course, and/or the use of an
alternative, second treatment); and/or
[0449] (x) administering the treatment (e.g., the first treatment
or a second (alternative) treatment) to the subject.
[0450] In one embodiment of the aforesaid methods, the subject is
identified as having decreased responsiveness to the treatment by
having at least one progression-positive biomarker. In one
embodiment, the progression-positive biomarker is a genomic
alteration in an NF-.kappa.B pathway gene. In an embodiment, the
progression-positive biomarker is a 6q deletion region, e.g., a
del(6q13-16) or a del(6q23-24). In one embodiment, the
progression-positive biomarker is a TNFAIP3 mutation or copy number
loss. In one embodiment, the progression-positive biomarker is an
EPHA7 mutation or copy number loss.
[0451] In one embodiment, the subject is a mammal, e.g., a human.
In one embodiment, the subject is at risk or suffers from a cancer
or tumor, e.g., a cancer or tumor described herein.
[0452] In one embodiment, the subject shows decreased
responsiveness to a PI3K inhibitor (e.g., is resistant or
refractive to treatment with a PI3K inhibitor, e.g., Compound
1).
[0453] In certain embodiments, the subject is identified as
developing resistance (e.g., acquired resistance) to the
monotherapy.
[0454] In one embodiment, the subject is identified as having a
decreased responsiveness (e.g., being resistance or having acquired
resistance) to a monotherapy treatment with a PI3K inhibitor (e.g.,
Compound 1 or GS1101) (referred herein to a "first PI3K inhibitor
treatment"). In one embodiment, the subject is identified as having
a decreased responsiveness (e.g., being resistance or having
acquired resistance) to a monotherapy treatment of a PI3K inhibitor
(e.g., Compound 1).
[0455] In one embodiment, the subject is identified as having an
increased responsiveness to a second treatment, e.g., a treatment
comprising a reduced dose of the PI3K inhibitor, or a treatment
comprising a combination of the PI3K inhibitor and a Bcl-2
inhibitor. In one embodiment, the dose of the PI3K inhibitor, the
Bcl-2 inhibitor, or both, is reduced, e.g., at least 20%, at least
30%, at least 40%, or at least 50%, than the amount or dosage of
each agent used individually, e.g., as a monotherapy.
[0456] In some embodiments of the methods described herein, the
method further includes altering a treatment (e.g., a first
treatment), a dose, a treatment schedule or time course, and/or the
use of an alternative, second treatment.
[0457] In other embodiments of the methods described herein, the
method further includes administering the treatment (e.g., the
first treatment or a second (alternative) treatment) to the
subject.
[0458] In other embodiments of the methods described herein, the
method further includes administering a combination of the PI3K
inhibitor and a Bcl-2 inhibitor in an amount sufficient to treat
the cancer, in the subject, e.g., for treatment of a cancer
described herein.
Detection of Alterations
[0459] The genomic alteration biomarkers provided herein can be
detected by the methods known in the art to detect genomic
alterations. In one embodiment, the gene mutations or copy number
alterations are detected by methods such as CytoScan Microarray
(pre- and post-treatment), targeted exome sequencing (pre- and
post-treatment), and Sanger sequencing. In one embodiment, the
mutation or copy number alteration of STK11 is detected by STK11
FISH Probe or qPCR.
[0460] In one embodiment, the biomarkers provided herein can be
used to identify, diagnose, predict efficacy, predict long term
clinical outcome, predict prognosis, and/or select patients for a
treatment described herein. In one embodiment, the biomarkers
provided herein can be used for subsets of patients with different
prognostic factors.
[0461] In the methods of the invention, one can detect expression
of biomarker proteins having at least one portion which is
displayed on the surface of tumor cells which express it. It is a
simple matter for the skilled artisan to determine whether a marker
protein, or a portion thereof, is exposed on the cell surface. For
example, immunological methods may be used to detect such proteins
on whole cells, or well known computer-based sequence analysis
methods may be used to predict the presence of at least one
extracellular domain (i.e. including both secreted proteins and
proteins having at least one cell-surface domain). Expression of a
marker protein having at least one portion which is displayed on
the surface of a cell which expresses it may be detected without
necessarily lysing the tumor cell (e.g. using a labeled antibody
which binds specifically with a cell-surface domain of the
protein).
[0462] Expression of a biomarkers described in this invention may
be assessed by any of a wide variety of well known methods for
detecting expression of a transcribed nucleic acid or protein.
Non-limiting examples of such methods include immunological methods
for detection of secreted, cell-surface, cytoplasmic, or nuclear
proteins, protein purification methods, protein function or
activity assays, nucleic acid hybridization methods, nucleic acid
reverse transcription methods, and nucleic acid amplification
methods.
[0463] In one embodiment, expression of a biomarker is assessed
using an antibody (e.g. a radio-labeled, chromophore-labeled,
fluorophore-labeled, or enzyme-labeled antibody), an antibody
derivative (e.g. an antibody conjugated with a substrate or with
the protein or ligand of a protein-ligand pair {e.g.
biotin-streptavidin}), or an antibody fragment (e.g. a single-chain
antibody, an isolated antibody hypervariable domain, etc.) which
binds specifically with a biomarker protein or fragment thereof,
including a biomarker protein which has undergone either all or a
portion of post-translational modifications to which it is normally
subjected in the tumor cell (e.g. glycosylation, phosphorylation,
methylation etc.).
[0464] In another embodiment, expression of a biomarker is assessed
by preparing mRNA/cDNA (i.e. a transcribed polynucleotide) from
cells in a patient sample, and by hybridizing the mRNA/cDNA with a
reference polynucleotide which is a complement of a biomarker
nucleic acid, or a fragment thereof. cDNA can, optionally, be
amplified using any of a variety of polymerase chain reaction
methods prior to hybridization with the reference polynucleotide.
Expression of one or more biomarkers can likewise be detected using
quantitative PCR to assess the level of expression of the
biomarker(s).
[0465] In all embodiments of the invention, the expression level of
a biomarker can be determined with reference to the effect on
biomarker expression caused by a mutation or variant in a gene
associated with said biomarker. Accordingly, for example, the
consequences of a genomic alteration on the expression level of
biomarkers referred to in the methods of the invention may be
inferred directly from identification of the genomic alteration in
the genome of a patient.
[0466] As used herein, the mutation can be a point mutation, e.g.
SNP, an insertion, a deletion, an amplification, a deletion, a
chromosomal translocation, an interstitial deletion, a chromosomal
inversion or a loss of heterozygosity.
[0467] In a related embodiment, a mixture of transcribed
polynucleotides obtained from the sample is contacted with a
substrate having fixed thereto a polynucleotide complementary to or
homologous with at least a portion (e.g. at least 7, 10, 15, 20,
25, 30, 40, 50, 100, 500, or more nucleotide residues) of a
biomarker nucleic acid. If polynucleotides complementary to or
homologous with are differentially detectable on the substrate
(e.g. detectable using different chromophores or fluorophores, or
fixed to different selected positions), then the levels of
expression of a plurality of biomarkers can be assessed
simultaneously using a single substrate (e.g. a "gene chip"
microarray of polynucleotides fixed at selected positions). When a
method of assessing biomarker expression is used which involves
hybridization of one nucleic acid with another, it is preferred
that the hybridization be performed under stringent hybridization
conditions.
[0468] When a plurality of biomarkers of the invention are used in
the methods of the invention, the level of expression of each
biomarker in a patient sample can be compared with the normal level
of expression of each of the plurality of biomarkers in
non-cancerous samples of the same type, either in a single reaction
mixture (i.e. using reagents, such as different fluorescent probes,
for each biomarker) or in individual reaction mixtures
corresponding to one or more of the biomarkers.
[0469] The level of expression of a biomarker in normal (i.e.
non-cancerous) human tissue can be assessed in a variety of ways.
In one embodiment, this normal level of expression is assessed by
assessing the level of expression of the biomarker in a portion of
cells which appears to be non-cancerous, and then comparing this
normal level of expression with the level of expression in a
portion of the tumor cells. Alternately, and particularly as
further information becomes available as a result of routine
performance of the methods described herein, population-average
values for normal expression of the biomarkers of the invention may
be used. In other embodiments, the `normal` level of expression of
a biomarker may be determined by assessing expression of the
biomarker in a patient sample obtained from a non-cancer-afflicted
patient, from a patient sample obtained from a patient before the
suspected onset of cancer in the patient, from archived patient
samples, and the like.
[0470] An exemplary method for detecting the presence or absence of
a biomarker protein or nucleic acid in a biological sample involves
obtaining a biological sample (e.g. a tumor-associated body fluid)
from a test patient and contacting the biological sample with a
compound or an agent capable of detecting the polypeptide or
nucleic acid (e.g., mRNA, genomic DNA, or cDNA). The detection
methods of the invention can thus be used to detect mRNA, protein,
cDNA, or genomic DNA, for example, in a biological sample in vitro
as well as in vivo. For example, in vitro techniques for detection
of mRNA include Northern hybridizations and in situ hybridizations.
In vitro techniques for detection of a biomarker protein include
enzyme linked immunosorbent assays (ELISAs), Western blots,
immunoprecipitations and immunofluorescence. In vitro techniques
for detection of genomic DNA include Southern hybridizations. In
vivo techniques for detection of mRNA include polymerase chain
reaction (PCR), Northern hybridizations and in situ hybridizations.
Furthermore, in vivo techniques for detection of a biomarker
protein include introducing into a patient a labeled antibody
directed against the protein or fragment thereof. For example, the
antibody can be labeled with a radioactive marker whose presence
and location in a patient can be detected by standard imaging
techniques.
[0471] A general principle of such diagnostic and prognostic assays
involves preparing a sample or reaction mixture that may contain a
biomarker, and a probe, under appropriate conditions and for a time
sufficient to allow the biomarker and probe to interact and bind,
thus forming a complex that can be removed and/or detected in the
reaction mixture. These assays can be conducted in a variety of
ways.
[0472] For example, one method to conduct such an assay would
involve anchoring the biomarker or probe onto a solid phase
support, also referred to as a substrate, and detecting target
biomarker/probe complexes anchored on the solid phase at the end of
the reaction. In one embodiment of such a method, a sample from a
patient, which is to be assayed for presence and/or concentration
of biomarker, can be anchored onto a carrier or solid phase
support. In another embodiment, the reverse situation is possible,
in which the probe can be anchored to a solid phase and a sample
from a patient can be allowed to react as an unanchored component
of the assay.
[0473] There are many established methods for anchoring assay
components to a solid phase. These include, without limitation,
biomarker or probe molecules which are immobilized through
conjugation of biotin and streptavidin. Such biotinylated assay
components can be prepared from biotin-NHS (N-hydroxy-succinimide)
using techniques known in the art (e.g., biotinylation kit, Pierce
Chemicals, Rockford, Ill.), and immobilized in the wells of
streptavidin-coated 96 well plates (Pierce Chemical). In certain
embodiments, the surfaces with immobilized assay components can be
prepared in advance and stored.
[0474] Other suitable carriers or solid phase supports for such
assays include any material capable of binding the class of
molecule to which the biomarker or probe belongs. Well-known
supports or carriers include, but are not limited to, glass,
polystyrene, nylon, polypropylene, nylon, polyethylene, dextran,
amylases, natural and modified celluloses, polyacrylamides,
gabbros, and magnetite.
[0475] In order to conduct assays with the above mentioned
approaches, the non-immobilized component is added to the solid
phase upon which the second component is anchored. After the
reaction is complete, uncomplexed components may be removed (e.g.,
by washing) under conditions such that any complexes formed will
remain immobilized upon the solid phase. The detection of
biomarker/probe complexes anchored to the solid phase can be
accomplished in a number of methods outlined herein.
[0476] In one embodiment, the probe, when it is the unanchored
assay component, can be labeled for the purpose of detection and
readout of the assay, either directly or indirectly, with
detectable labels discussed herein and which are well-known to one
skilled in the art.
[0477] It is also possible to directly detect biomarker/probe
complex formation without further manipulation or labeling of
either component (biomarker or probe), for example by utilizing the
technique of fluorescence energy transfer (i.e. FET, see for
example, Lakowicz et al., U.S. Pat. No. 5,631,169; Stavrianopoulos,
et al., U.S. Pat. No. 4,868,103). A fluorophore label on the first,
`donor` molecule is selected such that, upon excitation with
incident light of appropriate wavelength, its emitted fluorescent
energy will be absorbed by a fluorescent label on a second
`acceptor` molecule, which in turn is able to fluoresce due to the
absorbed energy. Alternately, the `donor` protein molecule may
simply utilize the natural fluorescent energy of tryptophan
residues. Labels are chosen that emit different wavelengths of
light, such that the `acceptor` molecule label may be
differentiated from that of the `donor`. Since the efficiency of
energy transfer between the labels is related to the distance
separating the molecules, spatial relationships between the
molecules can be assessed. In a situation in which binding occurs
between the molecules, the fluorescent emission of the `acceptor`
molecule label in the assay should be maximal. An FET binding event
can be conveniently measured through standard fluorometric
detection means well known in the art (e.g., using a
fluorimeter).
[0478] In another embodiment, determination of the ability of a
probe to recognize a biomarker can be accomplished without labeling
either assay component (probe or biomarker) by utilizing a
technology such as real-time Biomolecular Interaction Analysis
(BIA) (see, e.g., Sjolander, S. and Urbaniczky, C., 1991, Anal.
Chem. 63:2338-2345 and Szabo et al., 1995, Curr. Opin. Struct.
Biol. 5:699-705). As used herein, "BIA" or "surface plasmon
resonance" is a technology for studying biospecific interactions in
real time, without labeling any of the interactants (e.g.,
BIAcore). Changes in the mass at the binding surface (indicative of
a binding event) result in alterations of the refractive index of
light near the surface (the optical phenomenon of surface plasmon
resonance (SPR)), resulting in a detectable signal which can be
used as an indication of real-time reactions between biological
molecules.
[0479] Alternatively, in another embodiment, analogous diagnostic
and prognostic assays can be conducted with biomarker and probe as
solutes in a liquid phase. In such an assay, the complexed
biomarker and probe are separated from uncomplexed components by
any of a number of standard techniques, including but not limited
to: differential centrifugation, chromatography, electrophoresis
and immunoprecipitation. In differential centrifugation,
biomarker/probe complexes may be separated from uncomplexed assay
components through a series of centrifugal steps, due to the
different sedimentation equilibria of complexes based on their
different sizes and densities (see, for example, Rivas, G., and
Minton, A. P., 1993, Trends Biochem Sci. 18(8):284-7). Standard
chromatographic techniques may also be utilized to separate
complexed molecules from uncomplexed ones. For example, gel
filtration chromatography separates molecules based on size, and
through the utilization of an appropriate gel filtration resin in a
column format, for example, the relatively larger complex may be
separated from the relatively smaller uncomplexed components.
Similarly, the relatively different charge properties of the
biomarker/probe complex as compared to the uncomplexed components
may be exploited to differentiate the complex from uncomplexed
components, for example through the utilization of ion-exchange
chromatography resins. Such resins and chromatographic techniques
are well known to one skilled in the art (see, e.g., Heegaard, N.
H., 1998, J. Mol. Recognit. Winter 11(1-6):141-8; Hage, D. S., and
Tweed, S. A. J. Chromatogr B Biomed Sci Appl 1997 Oct. 10;
699(1-2):499-525). Gel electrophoresis may also be employed to
separate complexed assay components from unbound components (see,
e.g., Ausubel et al., ed., Current Protocols in Molecular Biology,
John Wiley & Sons, New York, 1987-1999). In this technique,
protein or nucleic acid complexes are separated based on size or
charge, for example. In order to maintain the binding interaction
during the electrophoretic process, non-denaturing gel matrix
materials and conditions in the absence of reducing agent are
typically preferred. Appropriate conditions to the particular assay
and components thereof will be well known to one skilled in the
art.
[0480] In a particular embodiment, the level of biomarker mRNA can
be determined both by in situ and by in vitro formats in a
biological sample using methods known in the art. The term
"biological sample" is intended to include tissues, cells,
biological fluids and isolates thereof, isolated from a patient, as
well as tissues, cells and fluids present within a patient. Many
expression detection methods use isolated RNA. For in vitro
methods, any RNA isolation technique that does not select against
the isolation of mRNA can be utilized for the purification of RNA
from tumor cells (see, e.g., Ausubel et al., ed., Current Protocols
in Molecular Biology, John Wiley & Sons, New York 1987-1999).
Additionally, large numbers of tissue samples can readily be
processed using techniques well known to those of skill in the art,
such as, for example, the single-step RNA isolation process of
Chomczynski (1989, U.S. Pat. No. 4,843,155).
[0481] The isolated mRNA can be used in hybridization or
amplification assays that include, but are not limited to, Southern
or Northern analyses, polymerase chain reaction analyses and probe
arrays. One preferred diagnostic method for the detection of mRNA
levels involves contacting the isolated mRNA with a nucleic acid
molecule (probe) that can hybridize to the mRNA encoded by the gene
being detected. The nucleic acid probe can be, for example, a
full-length cDNA, or a portion thereof, such as an oligonucleotide
of at least 7, 15, 30, 50, 100, 250 or 500 nucleotides in length
and sufficient to specifically hybridize under stringent conditions
to a mRNA or genomic DNA encoding a biomarker of the present
invention. Other suitable probes for use in the diagnostic assays
of the invention are described herein. Hybridization of an mRNA
with the probe indicates that the biomarker in question is being
expressed.
[0482] In one format, the mRNA is immobilized on a solid surface
and contacted with a probe, for example by running the isolated
mRNA on an agarose gel and transferring the mRNA from the gel to a
membrane, such as nitrocellulose. In an alternative format, the
probe(s) are immobilized on a solid surface and the mRNA is
contacted with the probe(s), for example, in an Affymetrix gene
chip array. A skilled artisan can readily adapt known mRNA
detection methods for use in detecting the level of mRNA encoded by
the biomarkers of the present invention.
[0483] An alternative method for determining the level of mRNA
biomarker in a sample involves the process of nucleic acid
amplification, e.g., by RT-PCR (the experimental embodiment set
forth in Mullis, 1987, U.S. Pat. No. 4,683,202), ligase chain
reaction (Barany, 1991, Proc. Natl. Acad. Sci. USA, 88:189-193),
self sustained sequence replication (Guatelli et al., 1990, Proc.
Natl. Acad. Sci. USA 87:1874-1878), transcriptional amplification
system (Kwoh et al., 1989, Proc. Natl. Acad. Sci. USA
86:1173-1177), Q-Beta Replicase (Lizardi et al., 1988,
Bio/Technology 6:1197), rolling circle replication (Lizardi et al.,
U.S. Pat. No. 5,854,033) or any other nucleic acid amplification
method, followed by the detection of the amplified molecules using
techniques well known to those of skill in the art. These detection
schemes are especially useful for the detection of nucleic acid
molecules if such molecules are present in very low numbers. As
used herein, amplification primers are defined as being a pair of
nucleic acid molecules that can anneal to 5' or 3' regions of a
gene (plus and minus strands, respectively, or vice-versa) and
contain a short region in between. In general, amplification
primers are from about 10 to 30 nucleotides in length and flank a
region from about 50 to 200 nucleotides in length. Under
appropriate conditions and with appropriate reagents, such primers
permit the amplification of a nucleic acid molecule comprising the
nucleotide sequence flanked by the primers.
[0484] For in situ methods, mRNA does not need to be isolated from
the tumor cells prior to detection. In such methods, a cell or
tissue sample is prepared/processed using known histological
methods. The sample is then immobilized on a support, typically a
glass slide, and then contacted with a probe that can hybridize to
mRNA that encodes the biomarker.
[0485] An alternative method for determining the level of mRNA
biomarker in a sample involves deep sequencing of cDNA generated
from RNA. In some embodiments, mRNA is isolated from tumor cells,
fragmented, and converted into cDNA libraries, and quantified using
next generation sequencing.
[0486] As an alternative to making determinations based on the
absolute expression level of the biomarker, determinations may be
based on the normalized expression level of the biomarker.
Expression levels are normalized by correcting the absolute
expression level of a biomarker by comparing its expression to the
expression of a gene that is not a biomarker, e.g., a housekeeping
gene that is constitutively expressed. Suitable genes for
normalization include housekeeping genes such as the actin gene, or
prognosis-positive cell-specific genes. This normalization allows
the comparison of the expression level in one sample, e.g., a
patient sample, to another sample, e.g., a non-tumor sample, or
between samples from different sources.
[0487] Alternatively, the expression level can be provided as a
relative expression level. To determine a relative expression level
of a biomarker (e.g. a prognosis-negative biomarker), the level of
expression of the biomarker is determined for 10 or more samples of
normal versus cancer cell isolates, preferably 50 or more samples,
prior to the determination of the expression level for the sample
in question. The mean expression level of each of the genes assayed
in the larger number of samples is determined and this is used as a
baseline expression level for the biomarker. The expression level
of the biomarker determined for the test sample (absolute level of
expression) is then divided by the mean expression value obtained
for that biomarker. This provides a relative expression level.
[0488] In another embodiment of the present invention, a biomarker
protein is detected. One agent for detecting biomarker protein of
the invention is an antibody capable of binding to such a protein
or a fragment thereof, preferably an antibody with a detectable
label. Antibodies can be polyclonal, or more preferably,
monoclonal. An intact antibody, or a fragment or derivative thereof
(e.g., Fab or F(ab')2 can be used. The term "labeled", with regard
to the probe or antibody, is intended to encompass direct labeling
of the probe or antibody by coupling (e.g., physically linking) a
detectable substance to the probe or antibody, as well as indirect
labeling of the probe or antibody by reactivity with another
reagent that is directly labeled. Examples of indirect labeling
include detection of a primary antibody using a fluorescently
labeled secondary antibody and end-labeling of a DNA probe with
biotin such that it can be detected with fluorescently labeled
streptavidin.
[0489] Proteins from tumor cells can be isolated using techniques
that are well known to those of skill in the art. The protein
isolation methods employed can, for example, be such as those
described in Harlow and Lane (Harlow and Lane, 1988, Antibodies: A
Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring
Harbor, N.Y.).
[0490] A variety of formats can be employed to determine whether a
sample contains a protein that binds to a given antibody. Examples
of such formats include, but are not limited to, enzyme immunoassay
(EIA), radioimmunoassay (RIA), Western blot analysis and enzyme
linked immunoabsorbant assay (ELISA). A skilled artisan can readily
adapt known protein/antibody detection methods for use in
determining whether tumor cells express a biomarker of the present
invention.
[0491] In one format, antibodies, or antibody fragments or
derivatives, can be used in methods such as Western blots or
immunofluorescence techniques to detect the expressed proteins. In
such uses, it is generally preferable to immobilize either the
antibody or proteins on a solid support. Suitable solid phase
supports or carriers include any support capable of binding an
antigen or an antibody. Well-known supports or carriers include
glass, polystyrene, polypropylene, polyethylene, dextran, nylon,
amylases, natural and modified celluloses, polyacrylamides,
gabbros, and magnetite.
[0492] One skilled in the art will know many other suitable
carriers for binding antibody or antigen, and will be able to adapt
such support for use with the present invention. For example,
protein isolated from tumor cells can be run on a polyacrylamide
gel electrophoresis and immobilized onto a solid phase support such
as nitrocellulose. The support can then be washed with suitable
buffers followed by treatment with the detectably labeled antibody.
The solid phase support can then be washed with the buffer a second
time to remove unbound antibody. The amount of bound label on the
solid support can then be detected by conventional means.
[0493] For ELISA assays, specific binding pairs can be of the
immune or non-immune type. Immune specific binding pairs are
exemplified by antigen-antibody systems or hapten/anti-hapten
systems. There can be mentioned fluorescein/anti-fluorescein,
dinitrophenyl/anti-dinitrophenyl, biotin/anti-biotin,
peptide/anti-peptide and the like. The antibody member of the
specific binding pair can be produced by customary methods familiar
to those skilled in the art. Such methods involve immunizing an
animal with the antigen member of the specific binding pair. If the
antigen member of the specific binding pair is not immunogenic,
e.g., a hapten, it can be covalently coupled to a carrier protein
to render it immunogenic. Non-immune binding pairs include systems
wherein the two components share a natural affinity for each other
but are not antibodies. Exemplary non-immune pairs are
biotin-streptavidin, intrinsic factor-vitamin B12, folic
acid-folate binding protein and the like.
[0494] A variety of methods are available to covalently label
antibodies with members of specific binding pairs. Methods are
selected based upon the nature of the member of the specific
binding pair, the type of linkage desired, and the tolerance of the
antibody to various conjugation chemistries. Biotin can be
covalently coupled to antibodies by utilizing commercially
available active derivatives. Some of these are
biotin-N-hydroxy-succinimide which binds to amine groups on
proteins; biotin hydrazide which binds to carbohydrate moieties,
aldehydes and carboxyl groups via a carbodiimide coupling; and
biotin maleimide and iodoacetyl biotin which bind to sulfhydryl
groups. Fluorescein can be coupled to protein amine groups using
fluorescein isothiocyanate. Dinitrophenyl groups can be coupled to
protein amine groups using 2,4-dinitrobenzene sulfate or
2,4-dinitrofluorobenzene. Other standard methods of conjugation can
be employed to couple monoclonal antibodies to a member of a
specific binding pair including dialdehyde, carbodiimide coupling,
homofunctional crosslinking, and heterobifunctional crosslinking
Carbodiimide coupling is an effective method of coupling carboxyl
groups on one substance to amine groups on another. Carbodiimide
coupling is facilitated by using the commercially available reagent
1-ethyl-3-(dimethyl-aminopropyl)-carbodiimide (EDAC).
[0495] Homobifunctional crosslinkers, including the bifunctional
imidoesters and bifunctional N-hydroxysuccinimide esters, are
commercially available and are employed for coupling amine groups
on one substance to amine groups on another. Heterobifunctional
crosslinkers are reagents which possess different functional
groups. The most common commercially available heterobifunctional
crosslinkers have an amine reactive N-hydroxysuccinimide ester as
one functional group, and a sulfhydryl reactive group as the second
functional group. The most common sulfhydryl reactive groups are
maleimides, pyridyl disulfides and active halogens. One of the
functional groups can be a photoactive aryl nitrene, which upon
irradiation reacts with a variety of groups.
[0496] The detectably-labeled antibody or detectably-labeled member
of the specific binding pair is prepared by coupling to a reporter,
which can be a radioactive isotope, enzyme, fluorogenic,
chemiluminescent or electrochemical materials. Two commonly used
radioactive isotopes are 125I and 3H. Standard radioactive isotopic
labeling procedures include the chloramine T, lactoperoxidase and
Bolton-Hunter methods for 125I and reductive methylation for 3H.
The term "detectably-labeled" refers to a molecule labeled in such
a way that it can be readily detected by the intrinsic enzymatic
activity of the label or by the binding to the label of another
component, which can itself be readily detected.
[0497] Enzymes suitable for use in this invention include, but are
not limited to, horseradish peroxidase, alkaline phosphatase,
.beta.-galactosidase, glucose oxidase, luciferases, including
firefly and renilla, .beta.-lactamase, urease, green fluorescent
protein (GFP) and lysozyme. Enzyme labeling is facilitated by using
dialdehyde, carbodiimide coupling, homobifunctional crosslinkers
and heterobifunctional crosslinkers as described above for coupling
an antibody with a member of a specific binding pair.
[0498] The labeling method chosen depends on the functional groups
available on the enzyme and the material to be labeled, and the
tolerance of both to the conjugation conditions. The labeling
method used in the present invention can be one of, but not limited
to, any conventional methods currently employed including those
described by Engvall and Pearlmann, Immunochemistry 8, 871 (1971),
Avrameas and Ternynck, Immunochemistry 8, 1175 (1975), Ishikawa et
al., J. Immunoassay 4(3):209-327 (1983) and Jablonski, Anal.
Biochem. 148:199 (1985).
[0499] Labeling can be accomplished by indirect methods such as
using spacers or other members of specific binding pairs. An
example of this is the detection of a biotinylated antibody with
unlabeled streptavidin and biotinylated enzyme, with streptavidin
and biotinylated enzyme being added either sequentially or
simultaneously. Thus, according to the present invention, the
antibody used to detect can be detectably-labeled directly with a
reporter or indirectly with a first member of a specific binding
pair. When the antibody is coupled to a first member of a specific
binding pair, then detection is effected by reacting the
antibody-first member of a specific binding complex with the second
member of the binding pair that is labeled or unlabeled as
mentioned above.
[0500] Moreover, the unlabeled detector antibody can be detected by
reacting the unlabeled antibody with a labeled antibody specific
for the unlabeled antibody. In this instance "detectably-labeled"
as used above is taken to mean containing an epitope by which an
antibody specific for the unlabeled antibody can bind. Such an
anti-antibody can be labeled directly or indirectly using any of
the approaches discussed above. For example, the anti-antibody can
be coupled to biotin which is detected by reacting with the
streptavidin-horseradish peroxidase system discussed above.
[0501] In one embodiment of this invention biotin is utilized. The
biotinylated antibody is in turn reacted with
streptavidin-horseradish peroxidase complex. Orthophenylenediamine,
4-chloro-naphthol, tetramethylbenzidine (TMB), ABTS, BTS or ASA can
be used to effect chromogenic detection.
[0502] In one immunoassay format for practicing this invention, a
forward sandwich assay is used in which the capture reagent has
been immobilized, using conventional techniques, on the surface of
a support. Suitable supports used in assays include synthetic
polymer supports, such as polypropylene, polystyrene, substituted
polystyrene, e.g. aminated or carboxylated polystyrene,
polyacrylamides, polyamides, polyvinylchloride, glass beads,
agarose, or nitrocellulose.
Kits
[0503] The invention also encompasses kits for detecting the
presence of a biomarker protein or nucleic acid in a biological
sample. Such kits can be used to determine if a patient is
suffering from or is at increased risk of developing a tumor that
is less susceptible to inhibition by PI3K inhibitors. For example,
the kit can comprise a labeled compound or agent capable of
detecting a biomarker protein or nucleic acid in a biological
sample and means for determining the amount of the protein or mRNA
in the sample (e.g., an antibody which binds the protein or a
fragment thereof, or an oligonucleotide probe which binds to DNA or
mRNA encoding the protein). Kits can also include instructions for
interpreting the results obtained using the kit.
[0504] For antibody-based kits, the kit can comprise, for example:
(1) a first antibody (e.g., attached to a solid support) which
binds to a biomarker protein; and, optionally, (2) a second,
different antibody which binds to either the protein or the first
antibody and is conjugated to a detectable label.
[0505] For oligonucleotide-based kits, the kit can comprise, for
example: (1) an oligonucleotide, e.g., a detectably labeled
oligonucleotide, which hybridizes to a nucleic acid sequence
encoding a biomarker protein or (2) a pair of primers useful for
amplifying a biomarker nucleic acid molecule. The kit can also
comprise, e.g., a buffering agent, a preservative, or a protein
stabilizing agent. The kit can further comprise components
necessary for detecting the detectable label (e.g., an enzyme or a
substrate). The kit can also contain a control sample or a series
of control samples which can be assayed and compared to the test
sample. Each component of the kit can be enclosed within an
individual container and all of the various containers can be
within a single package, along with instructions for interpreting
the results of the assays performed using the kit.
Dosing
[0506] In one embodiment, ABT-199, or a pharmaceutically acceptable
form thereof, is administered at a dosage of at an amount of less
than about 1000 mg, less than about 800 mg, less than about 750 mg,
less than about 500 mg, less than about 400 mg, less than about 350
mg, less than about 300 mg, less than about 250 mg, less than about
200 mg, less than about 150 mg, less than about 100 mg, less than
about 75 mg, less than about 50 mg, or less than about 25 mg daily.
In one embodiment, ABT-199, or a pharmaceutically acceptable form
thereof, is administered at a dosage of about 325 mg daily. In one
embodiment, ABT-199, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 250 mg daily. In one
embodiment, ABT-199, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 150 mg daily. In one
embodiment, ABT-199, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 100 mg, about 90 mg, about 85
mg, about 80 mg, about 75 mg, about 70 mg, about 65 mg, about 60
mg, about 55 mg, about 50 mg, about 45 mg, about 40 mg, about 35
mg, about 30 mg, about 25 mg, about 20 mg, about 15 mg, about 10
mg, or about 5 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 85 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 75 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 50 mg daily.
[0507] In one embodiment, ABT-263, or a pharmaceutically acceptable
form thereof, is administered at a dosage of at an amount of less
than about 1000 mg, less than about 800 mg, less than about 750 mg,
less than about 500 mg, less than about 400 mg, less than about 350
mg, less than about 300 mg, less than about 250 mg, less than about
200 mg, less than about 150 mg, less than about 100 mg, less than
about 75 mg, less than about 50 mg, or less than about 25 mg daily.
In one embodiment, ABT-263, or a pharmaceutically acceptable form
thereof, is administered at a dosage of about 325 mg daily. In one
embodiment, ABT-263, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 250 mg daily. In one
embodiment, ABT-263, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 150 mg daily. In one
embodiment, ABT-263, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 100 mg, about 90 mg, about 85
mg, about 80 mg, about 75 mg, about 70 mg, about 65 mg, about 60
mg, about 55 mg, about 50 mg, about 45 mg, about 40 mg, about 35
mg, about 30 mg, about 25 mg, about 20 mg, about 15 mg, about 10
mg, or about 5 mg daily. In one embodiment, ABT-263, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 85 mg daily. In one embodiment, ABT-263, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 75 mg daily. In one embodiment, ABT-263, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 50 mg daily.
[0508] In certain embodiments, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject a therapeutically
effective amount of a PI3K delta selective inhibitor (e.g.,
GS1101), or a pharmaceutically acceptable form thereof, and a Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof. In certain embodiments, the cancer is
diffuse large B-cell lymphoma (activated B-cell-like), diffuse
large B-cell lymphoma (germinal center B-cell-like), follicular
lymphoma, T-cell lymphoma, mantle cell lymphoma, multiple
myeloma.
[0509] In some embodiments of the methods described herein, the
PI3K delta inhibitor (e.g., GS1101), or a pharmaceutically
acceptable form thereof, and the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, are
administered at certain dosages. In one embodiment, provided herein
is a method of treating, managing, or preventing a cancer in a
subject comprising administering to the subject a therapeutically
effective amount of a PI3K delta inhibitor (e.g., GS1101), or a
pharmaceutically acceptable form thereof, and a Bcl-2 inhibitor, or
a pharmaceutically acceptable form thereof, wherein the PI3K delta
inhibitor (e.g., GS1101), or a pharmaceutically acceptable form
thereof, is administered at a dosage of in the range of from about
0.01 mg to about 75 mg daily and the Bcl-2 inhibitor (e.g., ABT-199
or ABT-263), or a pharmaceutically acceptable form thereof, is
administered at a dosage of in the range of from about 0.01 mg to
about 1100 mg daily.
[0510] In one embodiment, the PI3K delta inhibitor (e.g., GS1101),
or a pharmaceutically acceptable form thereof, is administered at a
dosage of in the range of from about 0.1 mg to about 500 mg, from
about 1 mg to about 500 mg, from about 10 mg to about 500 mg, from
about 50 mg to about 500 mg, from about 100 mg to about 400 mg,
from about 200 mg to about 400 mg, from about 250 mg to about 350
mg, or about 300 mg. In one embodiment, the composition comprises
the PI3K delta inhibitor (e.g., GS1101), or a pharmaceutically
acceptable form thereof, at an amount in the range of from about
0.1 mg to about 75 mg, from about 1 mg to about 75 mg, from about 5
mg to about 75 mg, from about 5 mg to about 60 mg, from about 5 mg
to about 50 mg, from about 5 mg to about 30 mg, from about 5 mg to
about 25 mg, from about 10 mg to about 25 mg, or from about 10 mg
to about 20 mg daily.
[0511] In one embodiment, the PI3K delta inhibitor (e.g., GS1101),
or a pharmaceutically acceptable form thereof, is administered at a
dosage of less than about 500 mg, less than about 400 mg, less than
about 350 mg, less than about 300 mg, less than about 250 mg, less
than about 200 mg, less than about 150 mg, less than about 100 mg,
less than about 75 mg, less than about 50 mg, less than about 30
mg, less than, less than about 25 mg, less than about 20 mg, less
than about 19 mg, less than about 18 mg, less than about 17 mg,
less than about 16 mg, less than about 16 mg, less than about 15
mg, less than about 14 mg, less than about 13 mg, less than about
12 mg, less than about 11 mg, or less than about 10 mg daily.
[0512] In certain embodiments, provided herein is a method of
treating, managing, or preventing a cancer in a subject comprising
administering to the subject a PI3K delta/gamma dual inhibitor, or
a pharmaceutically acceptable form thereof, in combination with a
Bcl-2 inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, wherein the cancer is diffuse large B-cell
lymphoma (activated B-cell-like), diffuse large B-cell lymphoma
(germinal center B-cell-like), follicular lymphoma, T-cell
lymphoma, mantle cell lymphoma, or multiple myeloma. In certain
embodiments,
[0513] In some embodiments of the methods described herein, the
PI3K delta/gamma dual inhibitor, or a pharmaceutically acceptable
form thereof, and the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263),
or a pharmaceutically acceptable form thereof, are administered at
certain dosages. In one embodiment, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject a PI3K delta/gamma
dual inhibitor, or a pharmaceutically acceptable form thereof, and
a Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof,
wherein the PI3K delta/gamma dual inhibitor, or a pharmaceutically
acceptable form thereof, is administered at a dosage of in the
range of from about 0.01 mg to about 75 mg daily and the Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, is administered at a dosage of in the
range of from about 0.01 mg to about 1100 mg daily.
[0514] In one embodiment, the PI3K delta/gamma dual inhibitor, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of in the range of from about 0.1 mg to about 75 mg, from
about 1 mg to about 75 mg, from about 5 mg to about 75 mg, from
about 5 mg to about 60 mg, from about 5 mg to about 50 mg, from
about 5 mg to about 30 mg, from about 5 mg to about 25 mg, from
about 10 mg to about 25 mg, or from about 10 mg to about 20 mg
daily.
[0515] In one embodiment, the PI3K delta/gamma dual inhibitor, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of less than about 25 mg, less than about 20 mg, less than
about 19 mg, less than about 18 mg, less than about 17 mg, less
than about 16 mg, less than about 16 mg, less than about 15 mg,
less than about 14 mg, less than about 13 mg, less than about 12
mg, less than about 11 mg, or less than about 10 mg daily.
[0516] In certain embodiments, provided herein is a method of
treating (e.g., inhibiting, managing, or preventing) a cancer in a
subject comprising administering to the subject a therapeutically
effective amount of Compound 1, or a pharmaceutically acceptable
form thereof, and a Bcl-2 inhibitor, or a pharmaceutically
acceptable form thereof, wherein the cancer is Diffuse large B-cell
lymphoma (activated B-cell-like), Diffuse large B-cell lymphoma
(germinal center B-cell-like), Follicular lymphoma, T-cell
lymphoma, Mantle cell lymphoma, Multiple myeloma. In one
embodiment, the Bcl-2 inhibitor is ABT-199. In another embodiment,
the Bcl-2 inhibitor is ABT-263.
[0517] In some embodiments of the methods described herein,
Compound 1, or a pharmaceutically acceptable form thereof, and the
Bcl-2 inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, are administered at certain dosages. In
one embodiment, provided herein is a method of treating, managing,
or preventing a cancer in a subject comprising administering to the
subject a therapeutically effective amount of Compound 1:
##STR00009##
or a pharmaceutically acceptable form thereof, and a Bcl-2
inhibitor, or a pharmaceutically acceptable form thereof, wherein
Compound 1, or a pharmaceutically acceptable form thereof, is
administered at a dosage of in the range of from about 0.01 mg to
about 75 mg daily and the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is
administered at a dosage of in the range of from about 0.01 mg to
about 1100 mg daily.
[0518] In one embodiment, Compound 1, or a pharmaceutically
acceptable form thereof, is administered at a dosage of in the
range of from about 0.1 mg to about 75 mg, from about 1 mg to about
75 mg, from about 5 mg to about 75 mg, from about 5 mg to about 60
mg, from about 5 mg to about 50 mg, from about 5 mg to about 30 mg,
from about 5 mg to about 25 mg, from about 10 mg to about 25 mg, or
from about 10 mg to about 20 mg daily.
[0519] In one embodiment, Compound 1, or a pharmaceutically
acceptable form thereof, is administered at a dosage of less than
about 25 mg, less than about 20 mg, less than about 19 mg, less
than about 18 mg, less than about 17 mg, less than about 16 mg,
less than about 16 mg, less than about 15 mg, less than about 14
mg, less than about 13 mg, less than about 12 mg, less than about
11 mg, or less than about 10 mg daily.
[0520] In one embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is
administered at a dosage of in the range of from about 0.1 mg to
about 800 mg, from about 0.1 mg to about 750 mg, from about 0.1 mg
to about 600 mg, from about 1 mg to about 500 mg, from about 1 mg
to about 400 mg, from about 10 mg to about 300 mg, or from about 50
mg to about 250 mg daily.
[0521] In one embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is
administered at a dosage of less than about 1000 mg, less than
about 800 mg, less than about 750 mg, less than about 500 mg, less
than about 400 mg, less than about 350 mg, less than about 300 mg,
less than about 250 mg, less than about 200 mg, less than about 150
mg, less than about 100 mg, less than about 75 mg, less than about
50 mg, or less than about 25 mg daily.
[0522] In one embodiment of the methods provided herein, ABT-199,
or a pharmaceutically acceptable form thereof, is administered at a
dosage of less than about 1000 mg, less than about 800 mg, less
than about 750 mg, less than about 500 mg, less than about 400 mg,
less than about 350 mg, less than about 300 mg, less than about 250
mg, less than about 200 mg, less than about 150 mg, less than about
100 mg, less than about 75 mg, less than about 50 mg, or less than
about 25 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 325 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 400 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 250 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 150 mg daily. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 100 mg daily, about 90 mg daily, about 85 mg daily,
about 80 mg daily, about 75 mg daily, about 70 mg daily, about 65
mg, about 60 mg daily, about 55 mg daily, about 50 mg daily, about
45 mg daily, about 40 mg daily, about 35 mg daily, about 30 mg
daily, about 25 mg daily, about 20 mg daily, about 15 mg daily,
about 10 mg daily, or about 5 mg daily. In one embodiment, ABT-199,
or a pharmaceutically acceptable form thereof, is administered at a
dosage of about 75 mg daily. In one embodiment, the composition
comprises ABT-199, or a pharmaceutically acceptable form thereof,
at an amount of about 55 mg. In one embodiment, ABT-199, or a
pharmaceutically acceptable form thereof, is administered at a
dosage of about 50 mg daily.
[0523] In one embodiment of the methods provided herein, ABT-263,
or a pharmaceutically acceptable form thereof, is administered at a
dosage of less than about 1000 mg, less than about 800 mg, less
than about 750 mg, less than about 500 mg, less than about 400 mg,
less than about 350 mg, less than about 325 mg, less than about 300
mg, less than about 250 mg, less than about 200 mg, less than about
150 mg, less than about 100 mg, less than about 75 mg, less than
about 50 mg, or less than about 25 mg daily. In one embodiment,
ABT-263, or a pharmaceutically acceptable form thereof, is
administered at a dosage of about 325 mg daily. In one embodiment,
ABT-263, or a pharmaceutically acceptable form thereof, is
administered at a dosage of about 250 mg daily. In one embodiment,
ABT-263, or a pharmaceutically acceptable form thereof, is
administered at a dosage of about 150 mg daily. In one embodiment,
ABT-263, or a pharmaceutically acceptable form thereof, is
administered at a dosage of about 100 mg daily, about 90 mg daily,
about 85 mg daily, about 80 mg daily, about 75 mg daily, about 70
mg daily, about 65 mg, about 60 mg daily, about 55 mg daily, about
50 mg daily, about 45 mg daily, about 40 mg daily, about 35 mg
daily, about 30 mg daily, about 25 mg daily, about 20 mg daily,
about 15 mg daily, about 10 mg daily, or about 5 mg daily. In one
embodiment, ABT-263, or a pharmaceutically acceptable form thereof,
is administered at a dosage of about 75 mg daily. In one
embodiment, the composition comprises ABT-263, or a
pharmaceutically acceptable form thereof, at an amount of about 55
mg. In one embodiment, ABT-263, or a pharmaceutically acceptable
form thereof, is administered at a dosage of about 50 mg daily.
[0524] As used herein, a daily dosage can be achieved by a single
administration of the targeted dosage amount or multiple
administrations of smaller dosage amount(s). For example, a 150 mg
daily dosage can be achieved by a single administration of 150 mg
of the therapeutic agent per day, two administrations of 75 mg of
the therapeutic agent per day, or three administrations of 50 mg of
the therapeutic agent per day.
[0525] In one embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), or a pharmaceutically acceptable form thereof, is
administered to the subject at least 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, 12 weeks, or 16 weeks before the
PI3K inhibitor (e.g., Compound 1), or a pharmaceutically acceptable
form thereof, is administered. In another embodiment, the Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), or a pharmaceutically
acceptable form thereof, is administered concurrently with the PI3K
inhibitor (e.g., Compound 1), or a pharmaceutically acceptable form
thereof, in a single dosage form or separate dosage forms. In yet
another embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263),
or a pharmaceutically acceptable form thereof, is administered to
the subject at least 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, 12 weeks, or 16 weeks after the PI3K inhibitor
(e.g., Compound 1), or a pharmaceutically acceptable form thereof,
is administered. In one embodiment, the Bcl-2 inhibitor is ABT-199.
In another embodiment, the Bcl-2 inhibitor is ABT-263.
[0526] In certain embodiments, the PI3K inhibitor (e.g., Compound
1), or a pharmaceutically acceptable form thereof, and the Bcl-2
inhibitor (e.g., ABT-199 or ABT-263), are administered via a same
route, e.g., both are administered orally. In other embodiments,
the PI3K inhibitor (e.g., Compound 1), or a pharmaceutically
acceptable form thereof, and the Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263), are administered via different routes, e.g., one is
administered orally and the other is administered intravenously. In
one embodiment, the PI3K inhibitor (e.g., Compound 1) is
administered orally once per day and ABT-199 is administered orally
once per day. In one embodiment, the PI3K inhibitor (e.g., Compound
1) is administered orally once per day and ABT-263 is administered
orally once per day.
[0527] Methods for monitoring minimal residual disease negativity
(MRD) are known in the art. See, e.g., Zhou, J. et al., Blood,
2007, 110: 1607-1611 (Prepublished online May 7, 2007. doi:
10.1182/blood-2006-09-045369). Such methods include DNA based tests
or RNA based tests. In certain embodiments, MRD is monitored using
flow cytometry, sequencing, or PCR.
[0528] In some embodiments, the compositions and methods described
herein are effective to reduce MRD.
[0529] In some embodiments, the methods described herein include
selecting a subject for treatment with the combination of a PI3K
inhibitor and a Bcl-2 inhibitor. In certain embodiments, the
subject (e.g., a patient with a cancer, e.g., a cancer described
herein) is selected for treatment with the combination based on the
MRD in the subject. In certain embodiments, the selection is based
on the presence of an MRD above a preselected level (e.g., 1
malignant cell in 100 normal cells, 1 malignant cell in 1000 normal
cells, or 1 malignant cell in 10,000 normal cells).
[0530] In some embodiments, the methods described herein further
comprise monitoring the MRD in a subject, e.g., evaluating MRD at
at least one, two, three, four, five, six, nine months after
initiating, continuing or ceasing treatment (e.g., PI3K inhibitor
monotherapy or Bcl-2 inhibitor monotherapy, or a combination
therapy disclosed herein).
[0531] In some embodiments, the combination of a PI3K inhibitor
(e.g. a PI3K inhibitor described herein) and a Bcl-2 inhibitor
(e.g., a Bcl-2 inhibitor described herein) is effective to reduce
the MRD in the subject, e.g., below a level previously measured in
the subject (e.g., the level measured before the combination
treatment). In certain embodiments, the combination of a PI3K
inhibitor and a Bcl-2 inhibitor is effective to reduce the MRD in
the subject below the level observed during or after treatment with
a monotherapy, e.g., a monotherapy comprising either the PI3K
inhibitor or the Bcl-2 inhibitor. In certain embodiments, the MRD
is decreased below the level observed during treatment with a
monotherapy comprising the PI3K inhibitor.
[0532] In certain embodiments, the combination is effective to
reduce the MRD below a preselected cutoff value (e.g., 1 malignant
cell in 100 normal cells, 1 malignant cell in 1000 normal cells, or
1 malignant cell in 10,000 normal cells). In certain embodiments,
the preselected cutoff value is 1 malignant cell in 1000 normal
cells. In those embodiments where the MRD is below a preselected
cutoff value (e.g., preselected cutoff value as described herein),
the treatment (e.g., PI3K inhibitor monotherapy or Bcl-2 inhibitor
monotherapy, or a combination therapy disclosed herein) can be
altered or discontinued. If upon monitoring the MRD (at at least
one, two, three, four, five, six, nine months after altering or
discontinuing the therapy), the MRD levels are increased above a
preselected cutoff (e.g., a preselected cutoff as described
herein), a second treatment can be initiated (e.g., PI3K inhibitor
monotherapy or Bcl-2 inhibitor monotherapy, a combination therapy
disclosed herein, or a combination with a third agent).
[0533] In some embodiments provided herein is a method of treating
cancer in a subject, the method comprising (i) administering to the
subject a monotherapy (e.g., a monotherapy comprising a PI3K
inhibitor or a second therapeutic agent as described herein) and
monitoring the MRD in the subject, and (ii) if the MRD increases
above a preselected cutoff value (e.g., 1 malignant cell in 100
normal cells, 1 malignant cell in 1000 normal cells, or 1 malignant
cell in 10,000 normal cells), administering to the subject a PI3K
inhibitor in combination with a Bcl-2 inhibitor. In certain
embodiments, the combination is effective to reduce the MRD, e.g.
to reduce the MRD below the cutoff value. In certain embodiments,
the preselected cutoff value is 1 malignant cell in 1000 or 10,000
normal cells.
[0534] In certain embodiments, provided herein is a method of
increasing the depth of response resulting in minimal residual
disease (MRD) negativity in a subject diagnosed with a cancer
comprising: (a) administering to the subject a therapeutically
effective amount of a PI3K inhibitor (e.g., Compound 1), and a
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof; (b)
monitoring the amount of MRD negativity in the subject, e.g., by
methods such as flow cytometry, sequencing, or PCR, and
administering a monotherapy comprising the PI3K inhibitor, or a
pharmaceutically acceptable form thereof, to the subject if the
amount of MRD negativity in the subject represents greater than 1
malignant cell in 1000 normal cells; and (c) monitoring the amount
of MRD negativity, e.g., by methods such as flow cytometry,
sequencing, or PCR in the subject receiving the monotherapy, and
administering a second combination therapy comprising the PI3K
inhibitor, or a pharmaceutically acceptable form thereof, and a
Bcl-2 inhibitor, or a pharmaceutically acceptable form thereof. In
one embodiment, the Bcl-2 inhibitor is ABT-199, ABT-263, ABT-737,
G3139 (genasense or oblimersen), GX15-070 (obatoclax mesylate),
HA14-1, TW-37, sabutoclax, Gossypol (AT-101), antimycin A,
apogossypol to the subject if the amount of MRD negativity in the
subject is at least 0.001%.
[0535] In certain embodiments, the combination therapy provided
herein leads to the reduction or diminishment of one or more
undesirable side effects associated with the monotherapy of either
the PI3K inhibitor or the Bcl-2 inhibitor. For example, monotherapy
of a Bcl-2 inhibitor (e.g., ABT-199 at a dose such as 1200 mg) can
cause tumor lysis syndrome. As such, a dose reduction of ABT-199
can diminish the undesirable side effects associated with
monotherapy of a Bcl-2 inhibitor.
[0536] A PI3K inhibitor and a Bcl-2 inhibitor can have synergistic
effects. The synergistic effect can be measured by timing, e.g., by
a delay in the development of resistance, to at least one of the
agents, or an increased in the period of remission. The synergistic
effect can also be measured in the quality of remission, e.g., the
synergistic effect can be a reduction in resistance (e.g., a
decrease in a measure of resistance or a decreased likelihood of
developing resistance), an increased likelihood of experiencing
complete remission, or a reduction in the levels of MRD (e.g., to
below detectable levels).
[0537] More specifically, provided herein (among other things) are
methods of delaying resistance of a subject, or prolonging
remission (e.g., complete remission or partial remission) of a
subject, to a PI3K inhibitor, e.g., Compound 1 or CAL-101, to a
Bcl-2 inhibitor, e.g., ABT-199 or ABT-263, or both of a PI3K
inhibitor and a Bcl-2 inhibitor. In some embodiments, the method
comprises administering a combination of a PI3K inhibitor (e.g.,
Compound 1 or CAL-101) and a Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263). Both agents may be administered to the subject before the
subject develops resistance to one or both of the PI3K inhibitor or
the Bcl-2 inhibitor, or after the subject develops resistance to
the PI3K inhibitor or the Bcl-2 inhibitor. In some embodiments,
once the subject becomes resistant to a first agent (e.g., the PI3K
inhibitor or Bcl-2 inhibitor), the first agent is withdrawn. In
other embodiments, once the subject becomes resistant to a first
agent (e.g., the PI3K inhibitor of Bcl-2 inhibitor), the first
agent is continued.
[0538] In some embodiments, the patient has elevated Bcl-2 levels,
e.g., in the cancer cell. In some embodiments, the patient has
elevated levels of a Bcl-2 family members such as NOXA, BMF, BIM,
PUMA, and HRK, e.g., in the cancer cell. Increased levels of Bcl-2
or its family member may be measured, e.g., by measuring DNA copy
number, RNA levels or protein levels. In some embodiments, the
level of Bcl-2 or its family member are increased by at least 25%,
50%, 75%, 2-fold, 3-fold, 5-fold, or 10-fold over levels before
treatment began (e.g., treatment with the PI3K inhibitor).
[0539] In certain embodiments, the subject is not resistant to a
PI3K inhibitor (e.g., Compound 1 or CAL-101). In some embodiments,
the subject is not resistant to a Bcl-2 inhibitor (e.g., ABT-199 or
ABT-263). In certain embodiments, the subject has previously been
administered a PI3K inhibitor (e.g., Compound 1 or CAL-101) as a
monotherapy or in combination with an agent other than a Bcl-2
inhibitor. In some embodiments, the subject has previously been
administered a Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) as a
monotherapy or in combination with an agent other than a PI3K
inhibitor. In some embodiments, the subject has a cancer, e.g., a
cancer described herein. In certain embodiments, in accordance with
the method, resistance is delayed compared to the time in which
resistance generally develops when the subject is treated with any
of the agents or inhibitors alone as monotherapy. In some
embodiments, the resistance is delayed by at least 2 weeks, e.g.,
at least 2 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5
months, 6 months, 8 months, 10 months, 12 months, 1 year, 2 years,
4 years, 6 years, 8 years, or more. In some embodiments, in
accordance with the method, remission (e.g., complete remission or
partial remission) is prolonged compared to the time in which
remission generally lasts when the subject is treated with any of
the agents or inhibitors alone as monotherapy. In certain
embodiments, remission (e.g., complete remission or partial
remission) is prolonged by at least 2 weeks, e.g., at least 2
weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6
months, 8 months, 10 months, 12 months, 1 year, 2 years, 4 years, 6
years, 8 years, or more.
[0540] Provided herein is also a method of reducing, e.g.,
overcoming resistance of a subject to a PI3K inhibitor (e.g.,
Compound 1 or CAL-101), comprising administering the PI3K inhibitor
as a monotherapy to the subject until development of resistance in
the subject to the PI3K inhibitor, and subsequently administering a
Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) to the subject. In some
cases, the method comprises continuing administration of the PI3K
inhibitor (e.g., at the same dosage, lower dosage, or higher
dosage) to the subject in combination with the Bcl-2 inhibitor. In
other cases, the method comprises discontinuing administration of
the PI3K inhibitor upon commencing administration of the Bcl-2
inhibitor. For example the administration of the PI3K inhibitor is
stopped before administration of the Bcl-2 inhibitor commences. In
other examples, the dosage of the PI3K inhibitor is decreased,
e.g., gradually, upon commencing administration of the Bcl-2
inhibitor.
4. Formulations
[0541] The formulations or compositions described herein can
include a PI3K inhibitor (e.g., one or more PI3K inhibitors as
described herein) and/or one or more additional agents (e.g., a
Bcl-2 inhibitor, e.g., one or more Bcl-2 inhibitors) as described
herein. In certain embodiments, the PI3K inhibitor (e.g., one or
more PI3K inhibitors as described herein) and the additional
agent(s) are included in the same dosage form. In certain
embodiments, the PI3K inhibitor (e.g., one or more PI3K inhibitors
as described herein) and the second agent (e.g., a Bcl-2 inhibitor,
e.g., one or more Bcl-2 inhibitors) are included in separate dosage
forms.
[0542] Pharmaceutical compositions may be specially formulated for
administration in solid or liquid form, including those adapted for
the following: oral administration, for example, drenches (aqueous
or non-aqueous solutions or suspensions), tablets (e.g., those
targeted for buccal, sublingual, and systemic absorption),
capsules, boluses, powders, granules, pastes for application to the
tongue, and intraduodenal routes; parenteral administration,
including intravenous, intraarterial, subcutaneous, intramuscular,
intravascular, intraperitoneal or infusion as, for example, a
sterile solution or suspension, or sustained-release formulation;
topical application, for example, as a cream, ointment, or a
controlled-release patch or spray applied to the skin;
intravaginally or intrarectally, for example, as a pessary, cream,
stent or foam; sublingually; ocularly; pulmonarily; local delivery
by catheter or stent; intrathecally, or nasally.
[0543] Examples of suitable aqueous and nonaqueous carriers which
may be employed in pharmaceutical compositions include water,
ethanol, polyols (such as glycerol, propylene glycol, polyethylene
glycol, and the like), and suitable mixtures thereof, vegetable
oils, such as olive oil, and injectable organic esters, such as
ethyl oleate. Proper fluidity may be maintained, for example, by
the use of coating materials, such as lecithin, by the maintenance
of the required particle size in the case of dispersions, and by
the use of surfactants.
[0544] These compositions can also contain adjuvants such as
preservatives, wetting agents, emulsifying agents, dispersing
agents, lubricants, and/or antioxidants. Prevention of the action
of microorganisms upon the compounds described herein may be
ensured by the inclusion of various antibacterial and antifungal
agents, for example, paraben, chlorobutanol, phenol sorbic acid,
and the like. It can also be desirable to include isotonic agents,
such as sugars, sodium chloride, and the like into the
compositions. In addition, prolonged absorption of the injectable
pharmaceutical form may be brought about by the inclusion of agents
which delay absorption such as aluminum monostearate and
gelatin.
[0545] Methods of preparing these formulations or compositions
include the step of bringing into association a compound described
herein and/or the chemotherapeutic with the carrier and,
optionally, one or more accessory ingredients. In general, the
formulations are prepared by uniformly and intimately bringing into
association a compound as disclosed herein with liquid carriers, or
finely divided solid carriers, or both, and then, if necessary,
shaping the product.
[0546] Preparations for such pharmaceutical compositions are
well-known in the art. See, e.g., Anderson, Philip O.; Knoben,
James E.; Troutman, William G, eds., Handbook of Clinical Drug
Data, Tenth Edition, McGraw-Hill, 2002; Pratt and Taylor, eds.,
Principles of Drug Action, Third Edition, Churchill Livingston,
N.Y., 1990; Katzung, ed., Basic and Clinical Pharmacology, Twelfth
Edition, McGraw Hill, 2011; Goodman and Gilman, eds., The
Pharmacological Basis of Therapeutics, Tenth Edition, McGraw Hill,
2001; Remingtons Pharmaceutical Sciences, 20th Ed., Lippincott
Williams & Wilkins, 2000; Martindale, The Extra Pharmacopoeia,
Thirty-Second Edition (The Pharmaceutical Press, London, 1999); all
of which are incorporated by reference herein in their entirety.
Except insofar as any conventional excipient medium is incompatible
with the compounds provided herein, such as by producing any
undesirable biological effect or otherwise interacting in a
deleterious manner with any other component(s) of the
pharmaceutically acceptable composition, the excipient's use is
contemplated to be within the scope of this disclosure.
[0547] In some embodiments, the concentration of the PI3K inhibitor
(e.g., one or more PI3K inhibitors, e.g., Compound 1 and/or GS1101)
or another agent (e.g., the Bcl 2 inhibitor, e.g., one or more Bcl
2 inhibitors as described herein) provided a pharmaceutical
composition disclosed herein or administered in a method disclosed
herein is less than about 100%, about 90%, about 80%, about 70%,
about 60%, about 50%, about 40%, about 30%, about 20%, about 19%,
about 18%, about 17%, about 16%, about 15%, about 14%, about 13%,
about 12%, about 11%, about 10%, about 9%, about 8%, about 7%,
about 6%, about 5%, about 4%, about 3%, about 2%, about 1%, about
0.5%, about 0.4%, about 0.3%, about 0.2%, about 0.1%, about 0.09%,
about 0.08%, about 0.07%, about 0.06%, about 0.05%, about 0.04%,
about 0.03%, about 0.02%, about 0.01%, about 0.009%, about 0.008%,
about 0.007%, about 0.006%, about 0.005%, about 0.004%, about
0.003%, about 0.002%, about 0.001%, about 0.0009%, about 0.0008%,
about 0.0007%, about 0.0006%, about 0.0005%, about 0.0004%, about
0.0003%, about 0.0002%, or about 0.0001%, w/w, w/v or v/v.
[0548] In some embodiments, the concentration of the PI3K inhibitor
(e.g., one or more PI3K inhibitors, e.g., Compound 1 and/or GS1101)
or another agent (e.g., the Bcl 2 inhibitor, e.g., one or more Bcl
2 inhibitors as described herein) provided a pharmaceutical
composition disclosed herein or administered in a method disclosed
herein is greater than about 90%, about 80%, about 70%, about 60%,
about 50%, about 40%, about 30%, about 20%, about 19.75%, about
19.50%, about 19.25%, about 19%, about 18.75%, about 18.50%, about
18.25%, about 18%, about 17.75%, about 17.50%, about 17.25%, about
17%, about 16.75%, about 16.50%, about 16.25%, about 16%, about
15.75%, about 15.50%, about 15.25%, about 15%, about 14.75%, about
14.50%, about 14.25%, about 14%, about 13.75%, about 13.50%, about
13.25%, about 13%, about 12.75%, about 12.50%, about 12.25%, about
12%, about 11.75%, about 11.50%, about 11.25%, about 11%, about
10.75%, about 10.50%, about 10.25%, about 10%, about 9.75%, about
9.50%, about 9.25%, about 9%, about 8.75%, about 8.50%, about
8.25%, about 8%, about 7.75%, about 7.50%, about 7.25%, about 7%,
about 6.75%, about 6.50%, about 6.25%, about 6%, about 5.75%, about
5.50%, about 5.25%, about 5%, about 4.75%, about 4.50%, about
4.25%, about 4%, about 3.75%, about 3.50%, about 3.25%, about 3%,
about 2.75%, about 2.50%, about 2.25%, about 2%, about 1.75%, about
1.50%, about 1.25%, about 1%, about 0.5%, about 0.4%, about 0.3%,
about 0.2%, about 0.1%, about 0.09%, about 0.08%, about 0.07%,
about 0.06%, about 0.05%, about 0.04%, about 0.03%, about 0.02%,
about 0.01%, about 0.009%, about 0.008%, about 0.007%, about
0.006%, about 0.005%, about 0.004%, about 0.003%, about 0.002%,
about 0.001%, about 0.0009%, about 0.0008%, about 0.0007%, about
0.0006%, about 0.0005%, about 0.0004%, about 0.0003%, about
0.0002%, or about 0.0001%, w/w, w/v, or v/v.
[0549] In some embodiments, the concentration of the PI3K inhibitor
(e.g., one or more PI3K inhibitors, e.g., Compound 1 and/or GS1101)
or another agent (e.g., the Bcl 2 inhibitor, e.g., one or more Bcl
2 inhibitors as described herein) provided a pharmaceutical
composition disclosed herein or administered in a method disclosed
herein is in the range from approximately 0.0001% to approximately
50%, approximately 0.001% to approximately 40%, approximately 0.01%
to approximately 30%, approximately 0.02% to approximately 29%,
approximately 0.03% to approximately 28%, approximately 0.04% to
approximately 27%, approximately 0.05% to approximately 26%,
approximately 0.06% to approximately 25%, approximately 0.07% to
approximately 24%, approximately 0.08% to approximately 23%,
approximately 0.09% to approximately 22%, approximately 0.1% to
approximately 21%, approximately 0.2% to approximately 20%,
approximately 0.3% to approximately 19%, approximately 0.4% to
approximately 18%, approximately 0.5% to approximately 17%,
approximately 0.6% to approximately 16%, approximately 0.7% to
approximately 15%, approximately 0.8% to approximately 14%,
approximately 0.9% to approximately 12%, or approximately 1% to
approximately 10%, w/w, w/v or v/v.
[0550] In some embodiments, the concentration of the PI3K inhibitor
(e.g., one or more PI3K inhibitors, e.g., Compound 1 and/or GS1101)
or another agent (e.g., the Bcl 2 inhibitor, e.g., one or more Bcl
2 inhibitors as described herein) provided a pharmaceutical
composition disclosed herein or administered in a method disclosed
herein is in the range from approximately 0.001% to approximately
10%, approximately 0.01% to approximately 5%, approximately 0.02%
to approximately 4.5%, approximately 0.03% to approximately 4%,
approximately 0.04% to approximately 3.5%, approximately 0.05% to
approximately 3%, approximately 0.06% to approximately 2.5%,
approximately 0.07% to approximately 2%, approximately 0.08% to
approximately 1.5%, approximately 0.09% to approximately 1%, or
approximately 0.1% to approximately 0.9%, w/w, w/v or v/v.
[0551] In some embodiments, the amount of Compound 1 or one or more
of the therapeutic agent disclosed herein is equal to or less than
about 10 g, about 9.5 g, about 9.0 g, about 8.5 g, about 8.0 g,
about 7.5 g, about 7.0 g, about 6.5 g, about 6.0 g, about 5.5 g,
about 5.0 g, about 4.5 g, about 4.0 g, about 3.5 g, about 3.0 g,
about 2.5 g, about 2.0 g, about 1.5 g, about 1.0 g, about 0.95 g,
about 0.9 g, about 0.85 g, about 0.8 g, about 0.75 g, about 0.7 g,
about 0.65 g, about 0.6 g, about 0.55 g, about 0.5 g, about 0.45 g,
about 0.4 g, about 0.35 g, about 0.3 g, about 0.25 g, about 0.2 g,
about 0.15 g, about 0.1 g, about 0.09 g, about 0.08 g, about 0.07
g, about 0.06 g, about 0.05 g, about 0.04 g, about 0.03 g, about
0.02 g, about 0.01 g, about 0.009 g, about 0.008 g, about 0.007 g,
about 0.006 g, about 0.005 g, about 0.004 g, about 0.003 g, about
0.002 g, about 0.001 g, about 0.0009 g, about 0.0008 g, about
0.0007 g, about 0.0006 g, about 0.0005 g, about 0.0004 g, about
0.0003 g, about 0.0002 g, or about 0.0001 g.
[0552] In some embodiments, the concentration of the PI3K inhibitor
(e.g., one or more PI3K inhibitors, e.g., Compound 1 and/or GS1101)
or another agent (e.g., the Bcl 2 inhibitor, e.g., one or more Bcl
2 inhibitors as described herein) provided a pharmaceutical
composition disclosed herein or administered in a method disclosed
herein is more than about 0.0001 g, about 0.0002 g, about 0.0003 g,
about 0.0004 g, about 0.0005 g, about 0.0006 g, about 0.0007 g,
about 0.0008 g, about 0.0009 g, about 0.001 g, about 0.0015 g,
about 0.002 g, about 0.0025 g, about 0.003 g, about 0.0035 g, about
0.004 g, about 0.0045 g, about 0.005 g, about 0.0055 g, about 0.006
g, about 0.0065 g, about 0.007 g, about 0.0075 g, about 0.008 g,
about 0.0085 g, about 0.009 g, about 0.0095 g, about 0.01 g, about
0.015 g, about 0.02 g, about 0.025 g, about 0.03 g, about 0.035 g,
about 0.04 g, about 0.045 g, about 0.05 g, about 0.055 g, about
0.06 g, about 0.065 g, about 0.07 g, about 0.075 g, about 0.08 g,
about 0.085 g, about 0.09 g, about 0.095 g, about 0.1 g, about 0.15
g, about 0.2 g, about 0.25 g, about 0.3 g, about 0.35 g, about 0.4
g, about 0.45 g, about 0.5 g, about 0.55 g, about 0.6 g, about 0.65
g, about 0.7 g, about 0.75 g, about 0.8 g, about 0.85 g, about 0.9
g, about 0.95 g, about 1 g, about 1.5 g, about 2 g, about 2.5 g,
about 3 g, about 3.5 g, about 4 g, about 4.5 g, about 5 g, about
5.5 g, about 6 g, about 6.5 g, about 7 g, about 7.5 g, about 8 g,
about 8.5 g, about 9 g, about 9.5 g, or about 10 g.
[0553] In some embodiments, the amount of Compound 1 or one or more
of the therapeutic agent disclosed herein is in the range of about
0.0001 to about 10 g, about 0.0005 to about 9 g, about 0.001 to
about 8 g, about 0.005 to about 7 g, about 0.01 to about 6 g, about
0.05 to about 5 g, about 0.1 to about 4 g, about 0.5 to about 4 g,
or about 1 to about 3 g.
Formulations for Oral Administration
[0554] In some embodiments of the methods described herein, PI3K
inhibitor (e.g., one or more PI3K inhibitors) and/or another agent
(e.g., the Bcl-2 inhibitor, e.g., one or more Bcl-2 inhibitors as
described herein) is administered orally. In certain embodiments of
the compositions described herein, PI3K inhibitor (e.g., Compound
1) and/or another agent (e.g., the Bcl-2 inhibitor, e.g., one or
more Bcl-2 inhibitors as described herein) is formulated for oral
administration. Some embodiments pertaining to such methods and
compositions include the following.
[0555] In some embodiments, provided herein are pharmaceutical
compositions for oral administration containing a compound as
disclosed herein, and a pharmaceutical excipient suitable for oral
administration. In some embodiments, provided herein are
pharmaceutical compositions for oral administration containing: (i)
an effective amount of a disclosed compound; optionally (ii) an
effective amount of one or more second agents; and (iii) one or
more pharmaceutical excipients suitable for oral administration. In
some embodiments, the pharmaceutical composition further contains:
(iv) an effective amount of a third agent.
[0556] In some embodiments, the pharmaceutical composition can be a
liquid pharmaceutical composition suitable for oral consumption.
Pharmaceutical compositions suitable for oral administration can be
presented as discrete dosage forms, such as capsules, cachets, or
tablets, or liquids or aerosol sprays each containing a
predetermined amount of an active ingredient as a powder or in
granules, a solution, or a suspension in an aqueous or non-aqueous
liquid, an oil-in-water emulsion, or a water-in-oil liquid
emulsion. Such dosage forms can be prepared by any of the methods
of pharmacy, but all methods include the step of bringing the
active ingredient into association with the carrier, which
constitutes one or more ingredients. In general, the pharmaceutical
compositions are prepared by uniformly and intimately admixing the
active ingredient with liquid carriers or finely divided solid
carriers or both, and then, if necessary, shaping the product into
the desired presentation. For example, a tablet can be prepared by
compression or molding, optionally with one or more accessory
ingredients. Compressed tablets can be prepared by compressing in a
suitable machine the active ingredient in a free-flowing form such
as powder or granules, optionally mixed with an excipient such as,
but not limited to, a binder, a lubricant, an inert diluent, and/or
a surface active or dispersing agent. Molded tablets can be made by
molding in a suitable machine a mixture of the powdered compound
moistened with an inert liquid diluent.
[0557] The present disclosure further encompasses anhydrous
pharmaceutical compositions and dosage forms comprising an active
ingredient, since water can facilitate the degradation of some
compounds. For example, water can be added (e.g., about 5%) in the
pharmaceutical arts as a means of simulating long-term storage in
order to determine characteristics such as shelf-life or the
stability of formulations over time. Anhydrous pharmaceutical
compositions and dosage forms can be prepared using anhydrous or
low moisture containing ingredients and low moisture or low
humidity conditions. For example, pharmaceutical compositions and
dosage forms which contain lactose can be made anhydrous if
substantial contact with moisture and/or humidity during
manufacturing, packaging, and/or storage is expected. An anhydrous
pharmaceutical composition can be prepared and stored such that its
anhydrous nature is maintained. Accordingly, anhydrous
pharmaceutical compositions can be packaged using materials known
to prevent exposure to water such that they can be included in
suitable formulary kits. Examples of suitable packaging include,
but are not limited to, hermetically sealed foils, plastic or the
like, unit dose containers, blister packs, and strip packs.
[0558] An active ingredient can be combined in an intimate
admixture with a pharmaceutical carrier according to conventional
pharmaceutical compounding techniques. The carrier can take a wide
variety of forms depending on the form of preparation desired for
administration. In preparing the pharmaceutical compositions for an
oral dosage form, any of the usual pharmaceutical media can be
employed as carriers, such as, for example, water, glycols, oils,
alcohols, flavoring agents, preservatives, coloring agents, and the
like in the case of oral liquid preparations (such as suspensions,
solutions, and elixirs) or aerosols; or carriers such as starches,
sugars, micro-crystalline cellulose, diluents, granulating agents,
lubricants, binders, and disintegrating agents can be used in the
case of oral solid preparations, in some embodiments without
employing the use of lactose. For example, suitable carriers
include powders, capsules, and tablets, with the solid oral
preparations. In some embodiments, tablets can be coated by
standard aqueous or nonaqueous techniques.
[0559] Binders suitable for use in pharmaceutical compositions and
dosage forms include, but are not limited to, corn starch, potato
starch, or other starches, gelatin, natural and synthetic gums such
as acacia, sodium alginate, alginic acid, other alginates, powdered
tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl
cellulose, cellulose acetate, carboxymethyl cellulose calcium,
sodium carboxymethyl cellulose), polyvinyl pyrrolidone, methyl
cellulose, pre-gelatinized starch, hydroxypropyl methyl cellulose,
microcrystalline cellulose, and mixtures thereof.
[0560] Examples of suitable fillers for use in the pharmaceutical
compositions and dosage forms disclosed herein include, but are not
limited to, talc, calcium carbonate (e.g., granules or powder),
microcrystalline cellulose, powdered cellulose, dextrates, kaolin,
mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch,
and mixtures thereof.
[0561] Disintegrants can be used in the pharmaceutical compositions
as provided herein to provide tablets that disintegrate when
exposed to an aqueous environment. Too much of a disintegrant can
produce tablets which can disintegrate in the bottle. Too little
can be insufficient for disintegration to occur and can thus alter
the rate and extent of release of the active ingredient(s) from the
dosage form. Thus, a sufficient amount of disintegrant that is
neither too little nor too much to detrimentally alter the release
of the active ingredient(s) can be used to form the dosage forms of
the compounds disclosed herein. The amount of disintegrant used can
vary based upon the type of formulation and mode of administration,
and can be readily discernible to those of ordinary skill in the
art. About 0.5 to about 15 weight percent of disintegrant, or about
1 to about 5 weight percent of disintegrant, can be used in the
pharmaceutical composition. Disintegrants that can be used to form
pharmaceutical compositions and dosage forms include, but are not
limited to, agar-agar, alginic acid, calcium carbonate,
microcrystalline cellulose, croscarmellose sodium, crospovidone,
polacrilin potassium, sodium starch glycolate, potato or tapioca
starch, other starches, pre-gelatinized starch, other starches,
clays, other algins, other celluloses, gums or mixtures
thereof.
[0562] Lubricants which can be used to form pharmaceutical
compositions and dosage forms include, but are not limited to,
calcium stearate, magnesium stearate, mineral oil, light mineral
oil, glycerin, sorbitol, mannitol, polyethylene glycol, other
glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated
vegetable oil (e.g., peanut oil, cottonseed oil, sunflower oil,
sesame oil, olive oil, corn oil, and soybean oil), zinc stearate,
ethyl oleate, ethylaureate, agar, or mixtures thereof. Additional
lubricants include, for example, a syloid silica gel, a coagulated
aerosol of synthetic silica, or mixtures thereof. A lubricant can
optionally be added, in an amount of less than about 1 weight
percent of the pharmaceutical composition.
[0563] When aqueous suspensions and/or elixirs are desired for oral
administration, the active ingredient therein can be combined with
various sweetening or flavoring agents, coloring matter or dyes
and, for example, emulsifying and/or suspending agents, together
with such diluents as water, ethanol, propylene glycol, glycerin
and various combinations thereof.
[0564] The tablets can be uncoated or coated by known techniques to
delay disintegration and absorption in the gastrointestinal tract
and thereby provide a sustained action over a longer period. For
example, a time delay material such as glyceryl monostearate or
glyceryl distearate can be employed. Formulations for oral use can
also be presented as hard gelatin capsules wherein the active
ingredient is mixed with an inert solid diluent, for example,
calcium carbonate, calcium phosphate or kaolin, or as soft gelatin
capsules wherein the active ingredient is mixed with water or an
oil medium, for example, peanut oil, liquid paraffin or olive
oil.
[0565] Surfactant which can be used to form pharmaceutical
compositions and dosage forms include, but are not limited to,
hydrophilic surfactants, lipophilic surfactants, and mixtures
thereof. That is, a mixture of hydrophilic surfactants can be
employed, a mixture of lipophilic surfactants can be employed, or a
mixture of at least one hydrophilic surfactant and at least one
lipophilic surfactant can be employed.
[0566] A suitable hydrophilic surfactant can generally have an HLB
value of at least about 10, while suitable lipophilic surfactants
can generally have an HLB value of or less than about 10. An
empirical parameter used to characterize the relative
hydrophilicity and hydrophobicity of non-ionic amphiphilic
compounds is the hydrophilic-lipophilic balance ("HLB" value).
Surfactants with lower HLB values are more lipophilic or
hydrophobic, and have greater solubility in oils, while surfactants
with higher HLB values are more hydrophilic, and have greater
solubility in aqueous solutions. Hydrophilic surfactants are
generally considered to be those compounds having an HLB value
greater than about 10, as well as anionic, cationic, or
zwitterionic compounds for which the HLB scale is not generally
applicable. Similarly, lipophilic (i.e., hydrophobic) surfactants
are compounds having an HLB value equal to or less than about 10.
However, HLB value of a surfactant is merely a rough guide
generally used to enable formulation of industrial, pharmaceutical
and cosmetic emulsions.
[0567] Hydrophilic surfactants can be either ionic or non-ionic.
Suitable ionic surfactants include, but are not limited to,
alkylammonium salts; fusidic acid salts; fatty acid derivatives of
amino acids, oligopeptides, and polypeptides; glyceride derivatives
of amino acids, oligopeptides, and polypeptides; lecithins and
hydrogenated lecithins; lysolecithins and hydrogenated
lysolecithins; phospholipids and derivatives thereof;
lysophospholipids and derivatives thereof; carnitine fatty acid
ester salts; salts of alkylsulfates; fatty acid salts; sodium
docusate; acylactylates; mono- and di-acetylated tartaric acid
esters of mono- and di-glycerides; succinylated mono- and
di-glycerides; citric acid esters of mono- and di-glycerides; and
mixtures thereof.
[0568] Within the aforementioned group, ionic surfactants include,
by way of example: lecithins, lysolecithin, phospholipids,
lysophospholipids and derivatives thereof; carnitine fatty acid
ester salts; salts of alkylsulfates; fatty acid salts; sodium
docusate; acylactylates; mono- and di-acetylated tartaric acid
esters of mono- and di-glycerides; succinylated mono- and
di-glycerides; citric acid esters of mono- and di-glycerides; and
mixtures thereof.
[0569] Ionic surfactants can be the ionized forms of lecithin,
lysolecithin, phosphatidylcholine, phosphatidylethanolamine,
phosphatidylglycerol, phosphatidic acid, phosphatidylserine,
lysophosphatidylcholine, lysophosphatidylethanolamine,
lysophosphatidylglycerol, lysophosphatidic acid,
lysophosphatidylserine, PEG-phosphatidylethanolamine,
PVP-phosphatidylethanolamine, lactylic esters of fatty acids,
stearoyl-2-lactylate, stearoyl lactylate, succinylated
monoglycerides, mono/diacetylated tartaric acid esters of
mono/diglycerides, citric acid esters of mono/diglycerides,
cholylsarcosine, caproate, caprylate, caprate, laurate, myristate,
palmitate, oleate, ricinoleate, linoleate, linolenate, stearate,
lauryl sulfate, teracecyl sulfate, docusate, lauroyl carnitines,
palmitoyl carnitines, myristoyl carnitines, and salts and mixtures
thereof.
[0570] Hydrophilic non-ionic surfactants can include, but are not
limited to, alkylglucosides; alkylmaltosides; alkylthioglucosides;
lauryl macrogolglycerides; polyoxyalkylene alkyl ethers such as
polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols such
as polyethylene glycol alkyl phenols; polyoxyalkylene alkyl phenol
fatty acid esters such as polyethylene glycol fatty acids
monoesters and polyethylene glycol fatty acids diesters;
polyethylene glycol glycerol fatty acid esters; polyglycerol fatty
acid esters; polyoxyalkylene sorbitan fatty acid esters such as
polyethylene glycol sorbitan fatty acid esters; hydrophilic
transesterification products of a polyol with at least one member
of glycerides, vegetable oils, hydrogenated vegetable oils, fatty
acids, and sterols; polyoxyethylene sterols, derivatives, and
analogues thereof; polyoxyethylated vitamins and derivatives
thereof; polyoxyethylene-polyoxypropylene block copolymers; and
mixtures thereof; polyethylene glycol sorbitan fatty acid esters
and hydrophilic transesterification products of a polyol with at
least one member of triglycerides, vegetable oils, and hydrogenated
vegetable oils. The polyol can be glycerol, ethylene glycol,
polyethylene glycol, sorbitol, propylene glycol, pentaerythritol,
or a saccharide.
[0571] Other hydrophilic-non-ionic surfactants include, without
limitation, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32
laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate, PEG-20
oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400
oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate,
PEG-100 stearate, PEG-20 dilaurate, PEG-25 glyceryl trioleate,
PEG-32 dioleate, PEG-20 glyceryl laurate, PEG-30 glyceryl laurate,
PEG-20 glyceryl stearate, PEG-20 glyceryl oleate, PEG-30 glyceryl
oleate, PEG-30 glyceryl laurate, PEG-40 glyceryl laurate, PEG-40
palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castor oil,
PEG-35 castor oil, PEG-60 castor oil, PEG-40 hydrogenated castor
oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6
caprate/caprylate glycerides, PEG-8 caprate/caprylate glycerides,
polyglyceryl-10 laurate, PEG-30 cholesterol, PEG-25 phyto sterol,
PEG-30 soya sterol, PEG-20 trioleate, PEG-40 sorbitan oleate,
PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9
lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl
ether, POE-20 stearyl ether, tocopheryl PEG-100 succinate, PEG-24
cholesterol, polyglyceryl-10 oleate, Tween 40, Tween 60, sucrose
monostearate, sucrose monolaurate, sucrose monopalmitate, PEG
10-100 nonyl phenol series, PEG 15-100 octyl phenol series, and
poloxamers.
[0572] Suitable lipophilic surfactants include, by way of example
only: fatty alcohols; glycerol fatty acid esters; acetylated
glycerol fatty acid esters; lower alcohol fatty acids esters;
propylene glycol fatty acid esters; sorbitan fatty acid esters;
polyethylene glycol sorbitan fatty acid esters; sterols and sterol
derivatives; polyoxyethylated sterols and sterol derivatives;
polyethylene glycol alkyl ethers; sugar esters; sugar ethers;
lactic acid derivatives of mono- and di-glycerides; hydrophobic
transesterification products of a polyol with at least one member
of glycerides, vegetable oils, hydrogenated vegetable oils, fatty
acids and sterols; oil-soluble vitamins/vitamin derivatives; and
mixtures thereof. Within this group, non-limiting examples of
lipophilic surfactants include glycerol fatty acid esters,
propylene glycol fatty acid esters, and mixtures thereof, or are
hydrophobic transesterification products of a polyol with at least
one member of vegetable oils, hydrogenated vegetable oils, and
triglycerides.
[0573] In one embodiment, the pharmaceutical composition can
include a solubilizer to ensure good solubilization and/or
dissolution of a compound as provided herein and to minimize
precipitation of the compound. This can be especially important for
pharmaceutical compositions for non-oral use, e.g., pharmaceutical
compositions for injection. A solubilizer can also be added to
increase the solubility of the hydrophilic drug and/or other
components, such as surfactants, or to maintain the pharmaceutical
composition as a stable or homogeneous solution or dispersion.
[0574] Examples of suitable solubilizers include, but are not
limited to, the following: alcohols and polyols, such as ethanol,
isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene
glycol, butanediols and isomers thereof, glycerol, pentaerythritol,
sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene
glycol, polypropylene glycol, polyvinylalcohol, hydroxypropyl
methylcellulose and other cellulose derivatives, cyclodextrins and
cyclodextrin derivatives; ethers of polyethylene glycols having an
average molecular weight of about 200 to about 6000, such as
tetrahydrofurfuryl alcohol PEG ether (glycofurol) or methoxy PEG;
amides and other nitrogen-containing compounds such as
2-pyrrolidone, 2-piperidone, .epsilon.-caprolactam,
N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone,
N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrolidone;
esters such as ethyl propionate, tributylcitrate, acetyl
triethylcitrate, acetyl tributyl citrate, triethylcitrate, ethyl
oleate, ethyl caprylate, ethyl butyrate, triacetin, propylene
glycol monoacetate, propylene glycol diacetate,
.epsilon.-caprolactone and isomers thereof, .delta.-valerolactone
and isomers thereof, .beta.-butyrolactone and isomers thereof; and
other solubilizers known in the art, such as dimethyl acetamide,
dimethyl isosorbide, N-methyl pyrrolidones, monooctanoin,
diethylene glycol monoethyl ether, and water.
[0575] Mixtures of solubilizers can also be used. Examples include,
but not limited to, triacetin, triethylcitrate, ethyl oleate, ethyl
caprylate, dimethylacetamide, N-methylpyrrolidone,
N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropyl
methylcellulose, hydroxypropyl cyclodextrins, ethanol, polyethylene
glycol 200-100, glycofurol, transcutol, propylene glycol, and
dimethyl isosorbide. In some embodiments, solubilizers include
sorbitol, glycerol, triacetin, ethyl alcohol, PEG-400, glycofurol
and propylene glycol.
[0576] The amount of solubilizer that can be included is not
particularly limited. The amount of a given solubilizer can be
limited to a bioacceptable amount, which can be readily determined
by one of skill in the art. In some circumstances, it can be
advantageous to include amounts of solubilizers far in excess of
bioacceptable amounts, for example to maximize the concentration of
the drug, with excess solubilizer removed prior to providing the
pharmaceutical composition to a subject using conventional
techniques, such as distillation or evaporation. Thus, if present,
the solubilizer can be in a weight ratio of about 10%, 25%, 50%,
100%, or up to about 200% by weight, based on the combined weight
of the drug, and other excipients. If desired, very small amounts
of solubilizer can also be used, such as about 5%, 2%, 1% or even
less. Typically, the solubilizer can be present in an amount of
about 1% to about 100%, more typically about 5% to about 25% by
weight.
[0577] The pharmaceutical composition can further include one or
more pharmaceutically acceptable additives and excipients. Such
additives and excipients include, without limitation, detackifiers,
anti-foaming agents, buffering agents, polymers, antioxidants,
preservatives, chelating agents, viscomodulators, tonicifiers,
flavorants, colorants, oils, odorants, opacifiers, suspending
agents, binders, fillers, plasticizers, lubricants, and mixtures
thereof.
[0578] Exemplary preservatives can include antioxidants, chelating
agents, antimicrobial preservatives, antifungal preservatives,
alcohol preservatives, acidic preservatives, and other
preservatives. Exemplary antioxidants include, but are not limited
to, alpha tocopherol, ascorbic acid, acorbyl palmitate, butylated
hydroxyanisole, butylated hydroxytoluene, monothioglycerol,
potassium metabisulfite, propionic acid, propyl gallate, sodium
ascorbate, sodium bisulfite, sodium metabisulfite, and sodium
sulfite. Exemplary chelating agents include
ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate,
disodium edetate, dipotassium edetate, edetic acid, fumaric acid,
malic acid, phosphoric acid, sodium edetate, tartaric acid, and
trisodium edetate. Exemplary antimicrobial preservatives include,
but are not limited to, benzalkonium chloride, benzethonium
chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium
chloride, chlorhexidine, chlorobutanol, chlorocresol,
chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine,
imidurea, phenol, phenoxyethanol, phenylethyl alcohol,
phenylmercuric nitrate, propylene glycol, and thimerosal. Exemplary
antifungal preservatives include, but are not limited to, butyl
paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic
acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate,
sodium benzoate, sodium propionate, and sorbic acid. Exemplary
alcohol preservatives include, but are not limited to, ethanol,
polyethylene glycol, phenol, phenolic compounds, bisphenol,
chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. Exemplary
acidic preservatives include, but are not limited to, vitamin A,
vitamin C, vitamin E, beta-carotene, citric acid, acetic acid,
dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.
Other preservatives include, but are not limited to, tocopherol,
tocopherol acetate, deteroxime mesylate, cetrimide, butylated
hydroxyanisol (BHA), butylated hydroxytoluened (BHT),
ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether
sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium
sulfite, potassium metabisulfite, Glydant Plus, Phenonip,
methylparaben, Germall 115, Germaben II, Neolone, Kathon, and
Euxyl. In certain embodiments, the preservative is an anti-oxidant.
In other embodiments, the preservative is a chelating agent.
[0579] Exemplary oils include, but are not limited to, almond,
apricot kernel, avocado, babassu, bergamot, black current seed,
borage, cade, camomile, canola, caraway, carnauba, castor,
cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton
seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol,
gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba,
kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut,
mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,
orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,
pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,
sasquana, savoury, sea buckthorn, sesame, shea butter, silicone,
soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut,
and wheat germ oils. Exemplary oils include, but are not limited
to, butyl stearate, caprylic triglyceride, capric triglyceride,
cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl
myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone
oil, and combinations thereof.
[0580] In addition, an acid or a base can be incorporated into the
pharmaceutical composition to facilitate processing, to enhance
stability, or for other reasons. Examples of pharmaceutically
acceptable bases include amino acids, amino acid esters, ammonium
hydroxide, potassium hydroxide, sodium hydroxide, sodium hydrogen
carbonate, aluminum hydroxide, calcium carbonate, magnesium
hydroxide, magnesium aluminum silicate, synthetic aluminum
silicate, synthetic hydrocalcite, magnesium aluminum hydroxide,
diisopropylethylamine, ethanolamine, ethylenediamine,
triethanolamine, triethylamine, triisopropanolamine,
trimethylamine, tris(hydroxymethyl)aminomethane (TRIS) and the
like. Also suitable are bases that are salts of a pharmaceutically
acceptable acid, such as acetic acid, acrylic acid, adipic acid,
alginic acid, alkanesulfonic acid, amino acids, ascorbic acid,
benzoic acid, boric acid, butyric acid, carbonic acid, citric acid,
fatty acids, formic acid, fumaric acid, gluconic acid,
hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic
acid, oxalic acid, para-bromophenylsulfonic acid, propionic acid,
p-toluenesulfonic acid, salicylic acid, stearic acid, succinic
acid, tannic acid, tartaric acid, thioglycolic acid,
toluenesulfonic acid, uric acid, and the like. Salts of polyprotic
acids, such as sodium phosphate, disodium hydrogen phosphate, and
sodium dihydrogen phosphate can also be used. When the base is a
salt, the cation can be any convenient and pharmaceutically
acceptable cation, such as ammonium, alkali metals, alkaline earth
metals, and the like. Examples can include, but not limited to,
sodium, potassium, lithium, magnesium, calcium and ammonium.
[0581] Suitable acids are pharmaceutically acceptable organic or
inorganic acids. Examples of suitable inorganic acids include
hydrochloric acid, hydrobromic acid, hydriodic acid, sulfuric acid,
nitric acid, boric acid, phosphoric acid, and the like. Examples of
suitable organic acids include acetic acid, acrylic acid, adipic
acid, alginic acid, alkanesulfonic acids, amino acids, ascorbic
acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric
acid, fatty acids, formic acid, fumaric acid, gluconic acid,
hydroquinosulfonic acid, isoascorbic acid, lactic acid, maleic
acid, methanesulfonic acid, oxalic acid, para-bromophenylsulfonic
acid, propionic acid, p-toluenesulfonic acid, salicylic acid,
stearic acid, succinic acid, tannic acid, tartaric acid,
thioglycolic acid, toluenesulfonic acid, uric acid and the
like.
[0582] In one embodiment, the therapeutic agent (e.g., Compound 1
or Bcl-2 inhibitor) can be formulated for oral dosing in phosal,
propylene glycol, polyethylene glycol, and ethanol. In one
embodiment, the Bcl-2 inhibitor (e.g., ABT-199 or ABT-263) can be
formulated for oral dosing in phosal, propylene glycol,
polyethylene glycol, and ethanol. For example, the formulation can
comprise of 60% phosal 50 propylene glycol (PG), 30% polyethylene
glycol (PEG) 400 and 10% ethanol.
5. Dosage
[0583] The PI3K inhibitor (e.g., Compound 1 or GS1101) or another
agent (e.g., a Bcl-2 inhibitor) disclosed herein may be delivered
in the form of pharmaceutically acceptable compositions which
comprise the PI3K inhibitor (e.g., (e.g., Compound 1 or GS1101)
described herein and/or one or more additional therapeutic agents,
formulated together with one or more pharmaceutically acceptable
excipients. In some instances, the PI3K inhibitor (e.g., Compound 1
or GS1101) or one or more of the other agents are administered in
separate pharmaceutical compositions and may (e.g., because of
different physical and/or chemical characteristics) be administered
by different routes (e.g., one therapeutic is administered orally,
while the other is administered intravenously). In other instances,
the PI3K inhibitor (e.g., Compound 1 or GS1101) or one or more of
the other agents disclosed herein may be administered separately,
but via the same route (e.g., both orally or both intravenously).
In still other instances, the PI3K inhibitor (e.g., Compound 1 or
GS1101) or one or more of the other agents disclosed herein may be
administered in the same pharmaceutical composition.
[0584] The selected dosage level will depend upon a variety of
factors including, for example, the activity of the particular
compound employed, the route of administration, the time of
administration, the rate of excretion or metabolism of the
particular compound being employed, the rate and extent of
absorption, the duration of the treatment, other drugs, compounds
and/or materials used in combination with the particular compound
employed, the age, sex, weight, condition, general health and prior
medical history of the patient being treated, and like factors well
known in the medical arts.
[0585] In general, a suitable daily dose of Compound 1 described
herein and/or a therapeutic agent will be that amount of the
compound which, in some embodiments, may be the lowest dose
effective to produce a therapeutic effect. Such an effective dose
will generally depend upon the factors described herein. Generally,
doses of Compound 1 or the therapeutic agent described herein for a
patient, when used for the indicated effects, will range from about
0.0001 mg to about 100 mg per day, or about 0.001 mg to about 100
mg per day, or about 0.01 mg to about 100 mg per day, or about 0.1
mg to about 100 mg per day, or about 0.0001 mg to about 500 mg per
day, or about 0.001 mg to about 500 mg per day, or about 0.01 mg to
1000 mg, or about 0.01 mg to about 500 mg per day, or about 0.1 mg
to about 500 mg per day, or about 1 mg to 50 mg per day, or about 5
mg to 40 mg per day. An exemplary dosage is about 10 to 30 mg per
day. In some embodiments, for a 70 kg human, a suitable dose would
be about 0.05 to about 7 g/day, such as about 0.05 to about 2.5
g/day. Actual dosage levels of the active ingredients in the
pharmaceutical compositions described herein may be varied so as to
obtain an amount of the active ingredient which is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient. In some instances, dosage levels below the lower limit of
the aforesaid range may be more than adequate, while in other cases
still larger doses may be employed without causing any harmful side
effect, e.g., by dividing such larger doses into several small
doses for administration throughout the day.
[0586] In some embodiments, the compounds may be administered
daily, every other day, three times a week, twice a week, weekly,
or bi-weekly. The dosing schedule can include a "drug holiday,"
e.g., the drug may be administered for two weeks on, one week off,
or three weeks on, one week off, or four weeks on, one week off,
etc., or continuously, without a drug holiday. The compounds may be
administered orally, intravenously, intraperitoneally, topically,
transdermally, intramuscularly, subcutaneously, intranasally,
sublingually, or by any other route.
[0587] In some embodiments, Compound 1 or the therapeutic agent
described herein may be administered in multiple doses. Dosing may
be about once, twice, three times, four times, five times, six
times, or more than six times per day. Dosing may be about once a
month, about once every two weeks, about once a week, or about once
every other day. In another embodiment, Compound 1 as disclosed
herein and another therapeutic agent are administered together from
about once per day to about 6 times per day. In another embodiment,
the administration of Compound 1 as provided herein and a
therapeutic agent continues for less than about 7 days. In yet
another embodiment, the administration continues for more than
about 6 days, about 10 days, about 14 days, about 28 days, about
two months, about six months, or about one year. In some cases,
continuous dosing is achieved and maintained as long as
necessary.
[0588] Administration of the pharmaceutical compositions as
disclosed herein may continue as long as necessary. In some
embodiments, an agent as disclosed herein is administered for more
than about 1, about 2, about 3, about 4, about 5, about 6, about 7,
about 14, or about 28 days. In some embodiments, an agent as
disclosed herein is administered for less than about 28, about 14,
about 7, about 6, about 5, about 4, about 3, about 2, or about 1
day. In some embodiments, a therapeutic agent as disclosed herein
is administered chronically on an ongoing basis, e.g., for the
treatment of chronic effects.
[0589] Since Compound 1 described herein may be administered in
combination with one or more therapeutic agent, the doses of each
agent or therapy may be lower than the corresponding dose for
single-agent therapy. The dose for single-agent therapy can range
from, for example, about 0.0001 to about 200 mg, or about 0.001 to
about 100 mg, or about 0.01 to about 100 mg, or about 0.1 to about
100 mg, or about 1 to about 50 mg per kilogram of body weight per
day.
[0590] When Compound 1 provided herein, is administered in a
pharmaceutical composition that comprises one or more therapeutic
agents, and the agent has a shorter half-life than Compound 1, unit
dose forms of the agent and Compound 1 may be adjusted
accordingly.
6. Kits
[0591] In some embodiments, provided herein are kits. The kits may
include a pharmaceutical composition as described herein, in
suitable packaging, and written material that can include
instructions for use, discussion of clinical studies, listing of
side effects, and the like. Such kits may also include information,
such as scientific literature references, package insert materials,
clinical trial results, and/or summaries of these and the like,
which indicate or establish the activities and/or advantages of the
pharmaceutical composition, and/or which describe dosing,
administration, side effects, drug interactions, or other
information useful to the health care provider. Such information
may be based on the results of various studies, for example,
studies using experimental animals involving in vivo models and
studies based on human clinical trials.
[0592] In some embodiments, a memory aid is provided with the kit,
e.g., in the form of numbers next to the tablets or capsules
whereby the numbers correspond with the days of the regimen which
the tablets or capsules so specified should be ingested. Another
example of such a memory aid is a calendar printed on the card,
e.g., as follows "First Week, Monday, Tuesday, . . . etc. . . .
Second Week, Monday, Tuesday, . . . " etc. Other variations of
memory aids will be readily apparent. A "daily dose" may be a
single tablet or capsule or several tablets or capsules to be taken
on a given day.
[0593] The kit may contain a PI3K inhibitor (e.g., one or more PI3K
inhibitors, e.g., Compound 1 or GS1101) in combination with another
agent (e.g., a Bcl-1 inhibitor, e.g., one or more Bcl-1 inhibitors
as described herein). In some embodiments, the PI3K inhibitor and
the other agent are provided as separate pharmaceutical
compositions in separate containers within the kit. In some
embodiments, the PI3K inhibitor as disclosed herein and the other
agent are provided as a single pharmaceutical composition within a
container in the kit. Suitable packaging and additional articles
for use (e.g., measuring cup for liquid preparations, foil wrapping
to minimize exposure to air, and the like) are known in the art and
may be included in the kit. In other embodiments, kits may further
comprise devices that are used to administer the active agents.
Examples of such devices include, but are not limited to, syringes,
drip bags, patches, and inhalers. Kits described herein may be
provided, marketed and/or promoted to health providers, including
physicians, nurses, pharmacists, formulary officials, and the like.
Kits can also, in some embodiments, be marketed directly to the
consumer.
[0594] An example of such a kit is a so-called blister pack.
Blister packs are well known in the packaging industry and are
being widely used for the packaging of pharmaceutical unit dosage
forms (tablets, capsules, and the like). Blister packs generally
consist of a sheet of relatively stiff material covered with a foil
of a preferably transparent plastic material. During the packaging
process, recesses are formed in the plastic foil. The recesses have
the size and shape of the tablets or capsules to be packed. Next,
the tablets or capsules are placed in the recesses and the sheet of
relatively stiff material is sealed against the plastic foil at the
face of the foil which is opposite from the direction in which the
recesses were formed. As a result, the tablets or capsules are
sealed in the recesses between the plastic foil and the sheet. The
strength of the sheet is such that the tablets or capsules may be
removed from the blister pack by manually applying pressure on the
recesses whereby an opening is formed in the sheet at the place of
the recess. The tablet or capsule can then be removed via said
opening.
[0595] Kits may further comprise pharmaceutically acceptable
vehicles that may be used to administer one or more active agents.
For example, if an active agent is provided in a solid form that
must be reconstituted for parenteral administration, the kit can
comprise a sealed container of a suitable vehicle in which the
active agent may be dissolved to form a particulate-free sterile
solution that is suitable for parenteral administration. Examples
of pharmaceutically acceptable vehicles include, but are not
limited to: Water for Injection USP; aqueous vehicles such as, but
not limited to, Sodium Chloride Injection, Ringer's Injection,
Dextrose Injection, Dextrose and Sodium Chloride Injection, and
Lactated Ringer's Injection; water-miscible vehicles such as, but
not limited to, ethyl alcohol, polyethylene glycol, and
polypropylene glycol; and non-aqueous vehicles such as, but not
limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl
oleate, isopropyl myristate, and benzyl benzoate.
[0596] The present disclosure further encompasses anhydrous
pharmaceutical compositions and dosage forms comprising an active
ingredient, since water can facilitate the degradation of some
compounds. For example, water may be added (e.g., about 5%) in the
pharmaceutical arts as a means of simulating long-term storage in
order to determine characteristics such as shelf-life or the
stability of formulations over time. Anhydrous pharmaceutical
compositions and dosage forms may be prepared using anhydrous or
low moisture containing ingredients and low moisture or low
humidity conditions. For example, pharmaceutical compositions and
dosage forms which contain lactose may be made anhydrous if
substantial contact with moisture and/or humidity during
manufacturing, packaging, and/or storage is expected. An anhydrous
pharmaceutical composition may be prepared and stored such that its
anhydrous nature is maintained. Accordingly, anhydrous
pharmaceutical compositions may be packaged using materials known
to prevent exposure to water such that they may be included in
suitable formulary kits. Examples of suitable packaging include,
but are not limited to, hermetically sealed foils, plastic or the
like, unit dose containers, blister packs, and strip packs.
EXAMPLES
Example 1
Combination Studies
[0597] The effect of the combination of Compound 1 and various
Bcl-2 inhibitors are studied using methods known in the art.
Provided below is an example where Compound 1 in combination with
one or more Bcl-2 inhibitor was examined in 20 cell lines.
[0598] The method is described as follows. Cells are thawed from a
liquid nitrogen preserved state. Once cells have been expanded and
divide at their expected doubling times, screening begins. Cells
are seeded in growth media in either black 1536-well or 384-well
tissue culture treated plates. Cells are then equilibrated in assay
plates via centrifugation and placed in incubators attached to the
Dosing Modules at 37.degree. C. for 24 hours before treatment. At
the time of treatment, a set of assay plates (which do not receive
treatment) are collected and ATP levels are measured by adding
ATPLite (Perkin Elmer). These Tzero (T.sub.0) plates are read using
ultra-sensitive luminescence on Envision plate readers (Perkin
Elmer). Treated assay plates are incubated with compound for 72
hours. After 72 hours, plates are developed for endpoint analysis
using ATPLite. All data points are collected via automated
processes, quality controlled and analyzed using Zalicus software.
Assay plates are accepted if they pass the following quality
control standards: relative luciferase values are consistent
throughout the entire experiment, Z-factor scores are greater than
0.6, untreated/vehicle controls behave consistently on the
plate.
[0599] Inhibition (I) is defined as
I=(1-T/V)*100%
where T is treated cell count and V is untreated (vehicle) cell
count (at 72 hours). I ranges from 0% (when T=V) to 100% (when
T=0). The IC.sub.50 value is defined as the drug concentration
needed to inhibit 50% of the cell growth compared to growth of the
vehicle treated cells (the drug concentration which gives I=50%).
The measure of effect in the experiment can be the inhibition of
cellular response relative to the untreated level (vehicle alone).
For untreated vehicle and treated levels V and T, a fractional
inhibition I=1-T/V is calculated. The inhibition ranges from 0% at
the untreated level to 100% when T=0. Inhibition levels are
negative for agents that actually increase levels. Other effect
measures, such as an activity ratio r=TN may be more appropriate
for some assays. When activity ratios (e.g., fold increase over
stimulated control) are being used, the effect can be measured
using an induction I=ln(T/V). With this definition, all effect
expressions are the same as for inhibition.
[0600] Growth Inhibition (GI) is used as a measure of cell
viability. The cell viability of vehicle is measured at the time of
dosing (T0) and after 72 hours (T72). A GI reading of 0% represents
no growth inhibition--T72 compound-treated and T72 vehicle signals
are matched. A GI reading of 100% represents complete growth
inhibition--T72 compound-treated and T0 vehicle signals are
matched. Cell numbers have not increased during the treatment
period in wells with GI 100% and may suggest a cytostatic effect
for compounds reaching a plateau at this effect level. A GI reading
of 200% represents complete death of all cells in the culture well.
Compounds reaching an activity plateau of GI 200% are considered
cytotoxic. GI is calculated by applying the following test and
equation:
If T < V 0 : 100 * ( 1 - T - V 0 V 0 ) ##EQU00001## If T
.gtoreq. V 0 : 100 * ( 1 - T - V 0 V - V 0 ) ##EQU00001.2##
where T is the signal measure for a test article, V is the
vehicle-treated control measure, and V.sub.0 is the vehicle control
measure at time zero. This formula is derived from the Growth
Inhibition calculation used in the National Cancer Institute's
NCI-60 high-throughput screen.
[0601] Combination analysis data were collected in a 6.times.6 dose
matrix. Synergy is calculated by comparing a combination's response
to those of its single compound, against the drug-with-itself
dose-additive reference model. Deviations from dose additivity may
be assessed visually on an isobologram or numerically with a
Combination Index (CI). See Tables below for CI at 50% inhibition
and CI at 50% growth inhibition. Additive effect is CI=1.0.
Synergistic effect is CI<1. Antagonistic effect is
CI>1.0.
[0602] Potency shifting was evaluated using an isobologram, which
demonstrates how much less drug is required in combination to
achieve a desired effect level, when compared to the single agent
doses needed to reach that effect. The isobologram was drawn by
identifying the locus of concentrations that correspond to crossing
the indicated inhibition level. This is done by finding the
crossing point for each single agent concentration in a dose matrix
across the concentrations of the other single agent. Practically,
each vertical concentration C.sub.Y is held fixed while a bisection
algorithm is used to identify the horizontal concentration C.sub.X
in combination with that vertical dose that gives the chosen effect
level in the response surface Z(C.sub.X,C.sub.Y). These
concentrations are then connected by linear interpolation to
generate the isobologram display. For synergistic interactions, the
isobologram contour fall below the additivity threshold and
approaches the origin, and an antagonistic interaction would lie
above the additivity threshold. The error bars represent the
uncertainty arising from the individual data points used to
generate the isobologram. The uncertainty for each crossing point
is estimated from the response errors using bisection to find the
concentrations where Z-.sigma..sub.Z(C.sub.X,C.sub.Y) and
Z+.sigma..sub.Z(C.sub.X,C.sub.Y) cross I.sub.cut, where
.sigma..sub.Z is the standard deviation of the residual error on
the effect scale.
[0603] To measure combination effects in excess of Loewe
additivity, a scalar measure to characterize the strength of
synergistic interaction termed the Synergy Score is devised. The
Synergy Score is calculated as:
Synergy Score=log f.sub.X log
f.sub.Y.SIGMA.max(0,I.sub.data)(I.sub.data-I.sub.Loewe)
The fractional inhibition for each component agent and combination
point in the matrix is calculated relative to the median of all
vehicle-treated control wells. The Synergy Score equation
integrates the experimentally-observed activity volume at each
point in the matrix in excess of a model surface numerically
derived from the activity of the component agents using the Loewe
model for additivity. Additional terms in the Synergy Score
equation (above) are used to normalize for various dilution factors
used for individual agents and to allow for comparison of synergy
scores across an entire experiment. The inclusion of positive
inhibition gating or an I.sub.data multiplier removes noise near
the zero effect level, and biases results for synergistic
interactions at that occur at high activity levels. Combinations
where the synergy score is greater than the mean self-cross plus
two standard deviations or three standard deviations can be
considered candidate synergies at 95% and 99% confidence levels,
respectively. Additivity should maintain a synergy score of zero,
and synergy score of two or three standard deviations indicate that
the combination is synergistic at statistically significant levels
of 95% and 99%.
[0604] The Synergy Score measure was used for the self-cross
analysis. Synergy Scores of self-crosses are expected to be
additive by definition and, therefore, maintain a synergy score of
zero. However, while some self-cross synergy scores are near zero,
many are greater suggesting that experimental noise or non-optimal
curve fitting of the single agent dose responses are contributing
to the slight perturbations in the score. This strategy was cell
line-centric, focusing on self-cross behavior in each cell line
versus a global review of cell line panel activity.
[0605] Loewe Volume (Loewe Vol) is used to assess the overall
magnitude of the combination interaction in excess of the Loewe
additivity model. Loewe Volume is particularly useful when
distinguishing synergistic increases in a phenotypic activity
(positive Loewe Volume) versus synergistic antagonisms (negative
Loewe Volume). When antagonisms are observed, as in the current
dataset, the Loewe Volume should be assessed to examine if there is
any correlation between antagonism and a particular drug
target-activity or cellular genotype. This model defines additivity
as a non-synergistic combination interaction where the combination
dose matrix surface should be indistinguishable from either drug
crossed with itself. The calculation for Loewe additivity is:
I.sub.Loewe that satisfies(X/X.sub.I)+(Y/Y.sub.I)=1
where XI and YI are the single agent effective concentrations for
the observed combination effect I. For example, if 50% inhibition
is achieved separately by 1 .mu.M of drug A or 1 .mu.M of drug B, a
combination of 0.5 .mu.M of A and 0.5 .mu.M of B should also
inhibit by 50%.
Results
[0606] The CI.sub.50 values for growth inhibition and inhibition in
Tables 1-6 are categorized as follows: S=0.1 to <0.5, T=0.5 to
<0.7, U=0.7 to <1, and W=.gtoreq.1. The synergy score values
for growth inhibition and inhibition are categorized as follows:
A1=0.0001 to <1, A2=1 to <3, A3=3 to <10, and
A4=.gtoreq.10.
[0607] The types of cell lines tested are diffuse large B-cell
lymphoma (DLBCL) activated B-cell-like (ABC), DLBCL germinal center
B-cell-like (GCB), follicular lymphoma, mantle cell lymphoma,
multiple myeloma, and T-cell lymphoma. These cell lines may have
different genomic profiles and thus, a combination of Compound 1
and a therapeutic agent can have different synergistic effects on
these cell lines. Data show that a combination of a gamma-selective
and a delta-selective compound provides a synergistic effect in
various types of cell lines.
Diffuse Large B-Cell Lymphoma (Activated B-Cell-Like)
[0608] Cell lines related to diffuse large B-cell lymphoma (DLBCL)
activated B-cell-like (ABC) were exposed to a combination of
Compound 1 and a Bcl-2 inhibitor. These cell lines include HBL-1,
OCI-Ly3, TMD8, and U2832. The results are shown in Table 1
below.
TABLE-US-00001 TABLE 1 Synergy score CI.sub.50 Synergy therapeutic
growth growth score CI.sub.50 agent Cell Line inhibition inhibition
inhibition inhibition ABT-199 HBL-1 A3 T A2 U ABT-199 OCI-Ly3 A3 S
A2 T ABT-199 U-2932 A3 S A2 S ABT-199 TMD8 A3 S A3 S ABT-263
OCI-Ly3 A3 U A1 W ABT-263 HBL-1 A3 S A2 U ABT-263 U-2932 A3 T A2 T
ABT-263 TMD8 A3 S A2 T
Diffuse Large B-Cell Lymphoma (Germinal Center B-Cell-Like)
[0609] Cell lines related to DLBCL germinal center B-cell-like
(GCB) were exposed to a combination of Compound 1 and a Bcl-2
inhibitor. These cell lines include DOHH-2, Farage, OCI-Ly7,
SU-DHL-10-epst, and SU-DHL-4-epst. For example, FIG. 1 (at 85%
growth inhibition) and FIG. 4 (at 180% growth inhibition) show
isobolograms depicting the synergistic effect of the combination of
Compound 1 and ABT-199 in SU-DHL-4 and DoHH-2 cell lines,
respectively. The results are shown in Table 2 below.
TABLE-US-00002 TABLE 2 Synergy score CI.sub.50 Synergy therapeutic
growth growth score CI.sub.50 agent Cell Line inhibition inhibition
inhibition inhibition ABT-199 OCI-Ly7 A1 A1 ABT-199 SU-DHL- A1 W A1
W 10-epst ABT-199 Farage A2 W A1 ABT-199 DOHH-2 A4 S A3 S ABT-199
SU-DHL- A4 S A3 S 4-epst ABT-263 OCI-Ly7 A1 W A1 ABT-263 Farage A3
T A2 T ABT-263 SU-DHL- A3 U A2 U 10-epst ABT-263 SU-DHL- A4 T A3 S
4-epst ABT-263 DOHH-2 A4 S A3 S
Follicular Lymphoma
[0610] Cell lines related to follicular lymphoma were exposed to a
combination of Compound 1 and a Bcl-2 inhibitor. These cell lines
include Karpas-422, RL, and WSU-NHL. For example, FIG. 3 (at 110%
growth inhibition) shows an isobologram depicting the synergistic
effect of the combination of Compound 1 and ABT-199 in WSU-NHL cell
line. The results are shown in Table 3 below.
TABLE-US-00003 TABLE 3 Synergy score CI.sub.50 Synergy therapeutic
growth growth score CI.sub.50 agent Cell Line inhibition inhibition
inhibition inhibition ABT-199 RL A3 U A1 W ABT-199 KARPAS- A3 T A2
U 422 ABT-199 WSU- A3 T A2 S NHL ABT-263 RL A3 U A1 U ABT-263 WSU-
A4 T A3 S NHL ABT-263 KARPAS- A4 S A3 T 422
T-Cell Lymphoma
[0611] Cell lines related to T-cell lymphoma were exposed to a
combination of Compound 1 and a Bcl-2 inhibitor. The cell line
includes HH and Karpas-299. The results are shown in Table 4
below.
TABLE-US-00004 TABLE 4 Synergy score CI.sub.50 Synergy therapeutic
Cell growth growth score CI.sub.50 agent Line inhibition inhibition
inhibition inhibition ABT-199 KARPAS-299 A1 A1 ABT-199 HH A2 S A1 T
ABT-263 KARPAS-299 A1 A1 ABT-263 HH A3 U A2 S
Mantle Cell Lymphoma
[0612] Cell lines related to mantle cell lymphoma were exposed to a
combination of Compound 1 and a Bcl-2 inhibitor. These cell lines
include GRANTA-519, Jeko-1 and Mino. For example, FIG. 2 (at 140%
growth inhibition) shows an isobologram depicting the synergistic
effect of the combination of Compound 1 and ABT-199 in Mino cell
line. The results are shown in Table 5 below.
TABLE-US-00005 TABLE 5 Synergy score CI.sub.50 Synergy therapeutic
Cell growth growth score CI.sub.50 agent Line inhibition inhibition
inhibition inhibition ABT-199 GRANTA-519 A2 T A1 W ABT-199 Jeko-1
A2 S A2 S ABT-199 Mino A4 S A3 S ABT-263 GRANTA-519 A3 S A1 W
ABT-263 Mino A3 S A2 T ABT-263 Jeko-1 A3 S A3 S
Multiple Myeloma
[0613] Cell lines related to multiple myeloma were exposed to a
combination of Compound 1 and a Bcl-2 inhibitor. These cell lines
include NCI-H929, OMP-2, and RPMI-8226. The results are shown in
Table 6 below.
TABLE-US-00006 TABLE 6 Synergy score CI.sub.50 Synergy therapeutic
Cell growth growth score CI.sub.50 agent Line inhibition inhibition
inhibition inhibition ABT-199 NCI-H929 A2 U A1 U ABT-199 RPMI-8226
A2 W A1 ABT-199 OPM-2 A2 U A1 ABT-263 NCI-H929 A2 U A2 T ABT-263
OPM-2 A3 T A2 U ABT-263 RPMI-8226 A3 T A2 U
Example 2
Gene Expression in Compound 1-Treated RR/CLL Subjects
[0614] Objectives of this experiment were to characterize the
mechanism of action of Compound 1 in CLL patients and support
rational combinations of Compound 1 with a second therapeutic
agent.
[0615] RNAseq technology was used to measure gene expression
changes in peripheral blood in patients treated with Compound 1.
The patients were treated as part of a clinical trial (identifier
NCT01476657) which is a phase 1 study in patients with advanced
hematologic malignancies. Briefly, RNA was extracted from 200 ul of
blood using TRI reagent, followed by RNA purification using a
Zymo-Spin column. Timepoints were taken at baseline and cycle 1 day
8 (C1D8), which is 7 days of Compound 1 treatment. Expression was
quantified using both FPKM and VOOM methods. Filters were used to
prioritize genes identified during this analysis.
[0616] The experiment indicated that about 500 genes exhibit
consistent changes across the population. The most notable pattern
was increased expression of multiple pro-apoptotic BH3 only genes
as well as increased expression of the anti-apoptotic gene Bcl-2.
Several Bcl-2 family genes were significantly changed. There were
also notable changes of gene expression in several B-cell receptor
pathway genes. Bcl These results show that BIK is downregulated,
while Bcl2, BMF, Bcl2L11 (also called BIM), PMAIP1 (also called
NOXA), BBC3 (also called PUMA), and HRK are up-regulated in
Compound 1-treated patients. Expression levels of selected genes
from this study are shown in FIGS. 5A, 5B, and 5C. FIG. 6 uses a
graph format to visualize several apoptotic genes examined in the
study, and illustrates increased levels of at least NOXA, BMF, BIM,
Bcl-22, PUMA, and HRK after 8 and 28 days after treatment with
Compound 1.
[0617] Levels of one of these Bcl-2 family members, HRK, are
explored in more detail in FIG. 7. In many of the patients having
stable disease (SD) or partial remission (PR), the HRK expression
level increased at least 1.5 fold after 7 days of Compound 1
treatment. These results indicate that increased HRK expression is
characteristic of patients that are treated with Compound 1. These
effects are seen broadly, in patients that respond to Compound 1
and patients that do not respond. These data suggests that cancer
cells respond to Compound 1 treatment by upregulating harakiri,
which pushes the balance of cell signaling towards apoptosis.
[0618] While not wishing to be bound by theory, this experiment
provides a rationale for a combination treatment with a PI3K
inhibitor such as Compound 1 with a Bcl-2 inhibitor. First,
increased expression of the pro-survival protein Bcl-2 can
contribute to resistance to PI3K inhibitor treatment. Accordingly,
administering a Bcl-2 inhibitor can overcome this resistance
mechanism by allowing the cell to enter the apoptosis pathway.
[0619] These data also indicates that cancer cells respond to
Compound 1 treatment by upregulating at least five pro-apoptotic
genes, e.g., inhibitors of Bcl-2, e.g., BIM, BMF, HRK, NOXA, and
PUMA. Without wishing to be bound by theory, this up-regulating of
pro-apoptotic genes can tip the balance of cell signaling towards
apoptosis. However, elevated levels of Bcl-2 can prevent these
pro-apoptotic factors from successfully initiating apoptosis. Thus,
by treating this cancer with a Bcl-2 inhibitor, one can unblock the
apoptosis pathway, leading to death of the cancer cell.
Furthermore, the elevated levels of pro-apoptotic factors suggests
that Compound 1-treated cells would be more sensitive than
untreated cells to a pro-apoptotic therapeutic such as a Bcl-2
inhibitor. Consequently, a combination therapy with a
PI3K-inhibitor such as Compound 1 and a Bcl-2 inhibitor can produce
unexpectedly high efficacy by acting in concert with high levels of
pro-apoptotic signaling factors, to trigger apoptosis.
[0620] In some embodiments, the methods of treatment described
herein comprise administering a combination of a PI3K inhibitor and
a modulator of, e.g., inhibitor of, the apoptotic signaling
pathway. Without being bound by theory, Compound 1 can enhance the
expression of key pro-apoptotic factors in CLL cells, but apoptosis
can be blocked. through a concomitant up-regulation of the
anti-apoptotic protein, Bcl-2. Inhibition of Bcl-2 by ABT-199 in
this setting can tip the balance toward apoptosis promoting tumor
cell death. The translational and ex-vivo experiments provided
herein support that a combination of Compound 1 and ABT-199 can
provide synergistic effects in subjects.
Example 3
Gene Expression in SU-DHL-4 DLBCL Cell Line
[0621] While Example 2 described the effects of Compound 1 on a
patient population including responders and non-responders, this
Example focuses on cells that are resistant to compound 1.
[0622] Experiments were performed to examine the pathway and gene
expression alterations in a cell line resistant to PI3K inhibitors.
SU-DHL-4 is a DLBCL cell line. SU-DHL-4 cells resistant to Compound
1 were generated by culturing the cells in the presence of Compound
1 or DMSO as a control for 8 weeks. Cells were subcloned under
selective pressure from the drug, seeding at densities of 3 cells
per well, 1 cell per well, or 0.3 cell per well. Parental,
DMSO-treated, and Compound 1-resistant clones were selected for
expansion. Five clones from each group were expanded. Cells were
harvested for various assays, including CellTiter-Glo.RTM. assays,
PD, RNA analysis, DNA analysis, and short tandem repeat (STR)
fingerprinting. A CTG assay was performed to confirm that cells
were resistant to Compound 1 at the time of sample collection,
whereas control cells (DMSO treated) were still resistant to
Compound 1.
[0623] Specific resistance was successfully created, as shown in
Table 7 below.
TABLE-US-00007 TABLE 7 AVG Cpnd 1 AVG IBR Clones IC50 (nM) IC50
(nM) Control 241 .+-. 17 447 .+-. 49 Cpnd 1 resistant 5420 .+-.
1079 697 .+-. 115 IBR resistant 805 .+-. 339 3531 .+-. 568
[0624] The gene expression pattern of Compound 1 resistant cell
clones was determined by RNASeq analysis, e.g., as described in
Wong et al. Nature Reviews Genetics 10.1(2009):57-63, incorporated
herein by reference. Briefly, FKPM values (Fragments Per Kilobase
Of Exon Per Million Fragments Mapped) were generated and filtered
by processing the raw values, normalizing, and filtering the
normalized values. This analysis revealed differential gene
expression unique and common to Compound 1-resistant and
ibrutinib-resistant clones. Specifically, 280 genes were
differentially regulated in the Compound 1-resistant cells, 190
genes were differentially regulated in the ibrutinib-resistant
cells, and 98 genes were differentially regulated in both the
Compound 1-resistant cells and the ibrutinib-resistant cells.
[0625] Next, the genes identified in this experiment were grouped
into pathways. 51 apoptotic pro-apoptotic genes (GeneGo Pathway ID
GO:0006915) were differentially regulated in the Compound
1-resistant cells. This observation support the idea that promoting
apoptosis (for instance, using a Bcl-2 inhibitor) can improve
cell-killing of Compound 1-resistant cells.
[0626] In this experiment, Bcl-2 was not strongly upregulated or
downregulated in the Compound 1-resistant cells. This result
indicates that Bcl-2 remains a viable drug target even in Compound
1-resistant cells. Without wishing to be bound by theory, this
finding supports the methods described herein involving treating a
cancer, such as a PI3K-resistant cancer, with a Bcl-2
inhibitor.
[0627] It was also observed that harakiri (HRK) mRNA levels
decreased in the Compound 1-resistant cells (as well as in
ibrutinib-resistant cells). While not wishing to be bound by
theory, this finding suggests that low levels of the pro-apoptotic
protein HRK contribute to Compound 1 resistance. This finding is in
agreement with the data in Example 2 showing that HRK expression
rises in Compound 1-treated patients, including patients who
respond to the treatment. Thus, high HRK expression is
characteristic of cancers that respond to Compound 1 treatment, and
low HRK expression is characteristic of at least a subset of
cancers that do not respond to Compound 1 treatment. This result
supports the rational combination of a PI3K inhibitor and a Bcl-2
inhibitor, for instance in patients that have a response (e.g., a
partial or complete response) to the PI3K inhibitor.
Example 4
STK11 Copy Number Loss in Patient with CLL
[0628] A patient diagnosed with CLL was treated by a monotherapy of
Compound 1 (25 mg bid) in a clinical trial. Serum samples of the
patient were collected at various points in the treatment. The copy
number of STK11 in the serum samples was determined by CytoScan
(Affymetrix). The results are described below:
[0629] At C1D1 (cycle 1, day 1), Absolute lymphocyte count
(ALC)=257, wild-type STK11 was detected;
[0630] At C3D1 (cycle 3, day 1), patient achieved partial
response;
[0631] At C5D1 (cycle 5, day 1), ALC=134, STK11 copy loss was
detected;
[0632] After C7 (cycle 7), patient progressed.
[0633] The result indicates that STK11 copy number loss can be
acquired and can be a contributing factor in acquired resistance to
the treatment of Compound 1.
Example 5
Genomic Profiling Protocol
[0634] Genomic DNA can be profiled by one or more of CytoScan
microarray analysis, targeted NexGen Sequencing and Sanger
Sequencing. The protocols for these methods are described herein.
CytoScan microarray analysis on genomic DNA can be used to
determine copy number alterations (CNAs), such as copy number loss
or gain. NexGen Sequencing on genomic DNA can be used to determine
gene mutations. Sanger sequencing on genomic DNA can be used to
determine IgHV mutation status. Results from genomic DNA profiling
were used to assess whether genomic alterations in individuals
treated with Compound 1 predict responsiveness or resistance to
treatment with Compound 1 and whether genomic alterations occur
with acquired resistance.
[0635] Preparation of DNA Sample
[0636] Peripheral whole blood samples were collected from CLL
patients being treated with Compound 1. Genomic DNA was extracted
from Cycle 1 Day 1 blood samples of 43 CLL patients, using QIAamp
DNA Blood Midi kit (Qiagen, cat #51185) according to the
manufacturer's protocol.
[0637] CytoScan Array Data Analysis
[0638] CytoScan array analysis allows for genome-wide
identification of copy number changes. The CytoScan HD array has
750,000 SNP probes and 1.9 million non-polymorphic probes,
providing even copy number coverage across the genome. The CytoScan
HD array also has intragenic coverage of 36,000 RefSeq genes.
[0639] Genomic DNA samples were applied for hybridization to
Affymetrix CytoScan HD arrays according to the manufacturer's
manual. CEL files were analyzed using Affymetrix software for
initial quality control, followed by the use of Nexus 7.5 software
(BioDiscovery, Inc.) for copy number and allelic analysis.
Following the profiling of copy number variations (CNVs) in each
sample, Nexus 7.5 software was used to identify the CNVs that are
significantly different between patients who responded to treatment
with Compound 1 and patients who did not (differential
frequency>25%; p<0.05). Copy number variances were initially
assessed with Nexus default setting (500 kb minimum LOH) for the
first set of 43 samples. In order to efficiently utilize allele
information, the segmentation window was changed to minimum LOH at
2 kb. Furthermore, gains that are not covered by an allelic event
were filtered out. The cancer-related genes were annotated based on
the Cancer Gene Census database. Association between CNVs and
clinical features were assessed by Fisher's exact test.
[0640] Targeted NexGen Sequencing and Data Analysis
[0641] Protocols for NexGen sequencing and hybrid capture are
described in Gnirke et al. (Nat Biotechnol. 27(2): 182-189, 2009).
In these experiments, hybrid capture approach was used with the
OncoGxOne leukemia/lymphoma panel (GeneWiz) containing 374 genes,
including all 4 PI3K isoforms, BTK, and PLC.gamma.. Illumina HiSeq
sequencing was used.
[0642] Agilent SureSelect solutions were used for the targeted DNA
capture of a panel of genes. According to the manufacturer's
protocol, DNA-Seq libraries were constructed and sequenced on
Illumina HiSeq 2500 using 100 bp paired-end reads. FASTQ files were
aligned by the OSA algorithm in Omicsoft Array Studio to generate
BAM files with default parameter setting. Non-synonymous mutations
including single-nucleotide variations (SNVs), insertions/deletions
(InDels) and stop codon gain/loss were detected by Array Studio's
mutation calling algorithm with the mutational allelic frequency
(MAF) threshold set to be above 0.1. Detected SNVs were annotated
with RefSeq gene model along with the Single Nucleotide
Polymorphism Database (dbSNP), Catalogue Of Somatic Mutations In
Cancer (COSMIC), and ClinVar databases to highlight the known
germline polymorphisms and the clinically relevant somatic
mutations. The putative somatic mutations were determined by
eliminating the SNVs that are known human single-nucleotide
polymorphisms (SNPs) archived in dbSNP and ClinVar and that were
detected in normal control samples. KEGG and MetoCore Pathway
Database was used to define the signaling pathways that are
significantly enriched with the genes that have somatic mutations
as detected in the CLL patients of this study (p<0.05).
Association between mutations and clinical features were assessed
by Fisher's exact test.
Example 6
Baseline Mutation Frequency in CLL
[0643] Using the targeted NexGen sequencing method described
previously, the baseline mutation frequency of CLL patients in the
Compound 1-treated patient population was determined, prior to
treatment of the patients with Compound 1 (Table 8). Many genes
that were previously described in the literature as being commonly
mutated in CLL were found to be mutated in the Compound 1-treated
population, suggesting that the Compound 1-treated population is
similar to what has been described for CLL (Landau et al. Cell 152,
714, 2013). The TP53 mutation rate was twice what has been
previously reported. This suggests that the Compound 1-treated
population has more aggressive disease than previously published
cohorts.
TABLE-US-00008 TABLE 8 Comparison of Compound 1-treated baseline
mutation frequency with literature. Compound 1-treated Landau et
al. (%) (%) Gene N = 160 N = 55 SF3B1 14 9 (5/55) TP53 13 24
(13/55) NOTCH1 10 20 (11/55) MYD88 8 5 (3/55) ATM 8 11 (6/55) XPO1
4 9 (5/55) POT1 3 0 NRAS 3 0 BCOR 3 0 KRAS 2 0 MED12 2 5 (3/55)
DDX3X 2 0 FBXW7 3 2 (1/55)
[0644] In addition, it was found that the average number of
baseline mutations per patient was relatively similar among
patients who show a complete or partial response to Compound 1
treatment, compared to non-responders (e.g., patients with stable
disease or progressive disease). The average number of baseline
mutations per patient was also relatively similar among R/R and
Tx-naive patients. Thus, the difference between a mutation profile
predictive of response and a mutation profile predictive of
non-response seems not to be the total number of mutations, but the
identity of the mutations.
Example 7
Baseline Copy Number Changes in CLL
[0645] Using the CytoScan array analysis, a genome-wide scan for
baseline copy number changes in the Compound 1-treated patient
population was performed, prior to treatment of the patients with
Compound 1. Copy number losses were in association with del(11q),
del(13q), and del(17p). In particular, genetic changes observed at
baseline included del(13q14), and del(11q22-23), del(17p13).
Del(8p) was also observed in the R/R population (6.5%) but not the
Tx-naive population. Also, copy number gain was observed in
association with trisomy 12. In summary, a copy gain at chromosome
12, trisomy 12; a copy loss at chromosome 11q22-23, del(11q22-23);
a copy loss at chromosome 13q14, del(13q14); and a copy loss at
chromosome 17p, del(17p) were observed.
Example 8
Copy Number Alterations in CLL
[0646] Using the CytoScan microarray analysis for genome wide as
described above, copy number alterations and losses of
heterozygosity were compared between responders and non-responders
to treatment with Compound 1. This analysis was performed in the
same CLL patient population as was assessed at baseline in Examples
6 and 7. Tumor response to drug is defined by SD/PD (Stable
Disease/Progressive Disease, i.e., non-responders) and CR/PR
(Complete Remission/Partial Remission, i.e., responders). Also
included in the responder group were PR patients with
lymphocytosis. See Brown, J. R. (2014) Blood, 123(22):3390-3397 and
Chesson, B. D. et al. Journal of Clinical Oncology,
30(23):2820-2822 for additional information regarding
classifications of patient responsiveness.
[0647] The genes for which differences between groups were
significant included BRAF, CTNNB1, FHIT, IRF4, MITF, MN1, NF2, RET,
STK11, TSC2, RB1, RANBP17, FGFR3, GMPS, and WHSC1. Summaries of
genetic alterations (500 kb minimum LOH) that were high in the
SD/PD group or low in the SD/PD group are provided in Tables 9 and
10 respectively.
TABLE-US-00009 TABLE 9 Summary of Changes Highin SD/PD group High
in Count of SD/PD Region Allelic 2 Imbalance CN Gain 30 CN Loss 37
LOH 66 Total 135
TABLE-US-00010 TABLE 10 Summary of Changes Lowin SD/PD groups Low
in Count of SD/PD Region CN Gain 6 CN Loss 6 LOH 56 Total 68
[0648] Table 11 shows copy number alterations for cancer genes with
a higher frequency in SD/PD (i.e., non-responder) patients compared
with CR/PR (i.e., responder) patients. BRAF, CTNNB1, FHIT, IRF4,
MITF, MN1, and NF2 had increased frequency of copy number gain in
SD/PD patients relative to CR/PR patients. NF2, RET, STK11, and
TSC2 had increased frequency of copy number loss in SD/PD patients
relative to CR/PR patients. RB1 showed a higher frequency of loss
of heterozygosity in SD/PD patients relative to CR/PR patients.
[0649] The results presented in Table 10 indicate that copy number
gain in each of BRAF, CTNNB1, FHIT, IRF4, MITF, MN1, and NF2 is
associated with or predictive of nonresponsiveness or resistance
(e.g., acquired resistance) of a cancer (e.g., a CLL) to a PI3K
inhibitor (e.g., Compound 1). The results presented in Table 10
also indicate that copy number loss in each of NF2, RET, STK11, and
TSC2 is associated with or predictive of nonresponsiveness or
resistance (e.g., acquired resistance) of a cancer (e.g., a CLL) to
a PI3K inhibitor (e.g., Compound 1). The results presented in Table
10 further suggest loss of heterozygosity in RB1 is associated with
or predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor (e.g.,
Compound 1).
TABLE-US-00011 TABLE 11 Cancer genes with higher frequency in SD/PD
CN gain CN loss LOH BRAF NF2 RB1 CTNNB1 RET FHIT STK11 IRF4 TSC2
MITF MN1 NF2
[0650] Table 12 shows copy number alterations for cancer genes with
a lower frequency in SD/PD patients compared with CR/PR patients.
Copy number gain in RANBP17 had a lower frequency in SD/PD (i.e.,
non-responder) patients compared with CR/PR (i.e., responder)
patients. Also, loss of heterozygosity in FGFR3, GMPS, and WHSC1
had a lower frequency in SD/PD (i.e., non-responder) patients
compared with CR/PR (i.e., responder) patients.
[0651] These results presented in Table 12 indicate that copy
number gain in RANBP17 is associated with responsiveness or lack of
resistance (e.g., acquired resistance) of a cancer (e.g., a CLL) to
a PI3K inhibitor (e.g., Compound 1). These results presented in
Table 12 also indicate that loss of heterozygosity in each of
FGFR3, GMPS, and WHSC1 is associated with or predictive of
responsiveness or lack of resistance (e.g., acquired resistance) of
a cancer (e.g., a CLL) to a PI3K inhibitor (e.g., Compound 1).
TABLE-US-00012 TABLE 12 Cancer genes w/Lower frequency in SD/PD CN
gain CN loss LOH RANBP17 FGFR3 GMPS WHSC1
[0652] In order to get more specific LOH calls and increase
confidence of copy number calling, the CNV data was analyzed with a
different segmentation window (minimum LOH is 2 kb).
[0653] Table 13 shows copy number alterations for cancer genes with
a higher frequency of loss in SD/PD patients compared with CR/PR
patients (>25% frequency difference, p<0.05). Loss of
CBFA2T3, YWHAE, TP53, PER1 and GAS7 are accompanied with an allelic
event (allele imbalance or loss of heterozygosity); while only copy
number loss was found in STK11, FSTL3 and USP6. Among all patients,
loss of YWHAE, STK11, TP53, FSTL3 and USP6 are significantly more
frequent in SD/PD patients compared with CR/PR patients. Within the
refractory/relapsed cohort (R/R), loss of STK11, TP53, PER1, GAS7
and FSTL3 occur more significantly in SD/PD patients compared to
CR/PR patients.
[0654] The results presented in Table 13 indicate loss of YWHAE,
STK11, TP53, FSTL3 and USP6 are associated with or predictive of
nonresponsiveness or resistance (e.g., acquired resistance) of a
cancer (e.g., a CLL) for all patients to a PI3K inhibitor (e.g.,
Compound 1). The results presented in Table 13 further suggest loss
of STK11, TP53, PER1, GAS7 and FSTL3 is associated with or
predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) among refractory or relapsed
patients to a PI3K inhibitor (e.g., Compound 1).
[0655] Table 14 shows copy number alterations for cancer genes with
a differential frequency of loss in nodal responders compared to
nodal nonresponders (>25% frequency difference, p<0.05). For
these three cancer genes, copy number loss was identified without
coverage of an allelic event. TSC1 and NF2 are more frequently loss
in nodal nonresponders compared to nodal responders, whereas EGFR
loss is found significantly frequently lost in nodal
responders.
[0656] The results presented in Table 14 indicate that loss of EGFR
is associated with or predictive of responsiveness or lack of
resistance (e.g., acquired resistance) of a cancer (e.g., a CLL)
for all patients to a PI3K inhibitor (e.g., Compound 1).
TABLE-US-00013 TABLE 13 Cancer genes w/higher frequency in SD/PD
Fisher's Fisher's exact exact Allelic (all patients (R/R only Loss
Chr Event n = 56) n = 46) CBFA2T3 16q24 Yes 0.174 0.1378 YWHAE
17p13 Yes 0.0459* 0.0626 STK11 19p13 0.0459* 0.0042** TP53 17p13
Yes 0.0371* 0.0274* PERI 17p13 Yes 0.0696 0.0274* GAS7 17p13 Yes
0.0696 0.0274* FSTL3 19p13 0.006** 0.0022** USP6 17p13 0.0459*
0.0626 MAP2K4 17p12 0.0696 0.0274*
TABLE-US-00014 TABLE 14 Cancer genes w/differential frequency
between nodal responders and nodal nonresponders Loss Higher
Frequency Fisher's exact TSC1 Non-responder 0.09 NF2 Non-responder
0.057 EGFR Responder 0.035*
[0657] Validation of copy number losses of several genes was
performed by RNAseq, e.g., as described in Wong et al. Nature
Reviews Genetics 10.1(2009):57-63, incorporated herein by
reference. The relative expression levels of TP53, YWHAE, and STK11
are reduced in patients having a loss in copy number, compared to
patients with no loss in copy number, as shown in FIGS. 13A, 13B,
and 13C.
Example 10
Relationship Between Mutational and Copy Number Variation
Frequencies and Responses
[0658] The relationship between certain genetic alterations (e.g.,
exonic deletions) and patient responsiveness to Compound 1 was
analyzed.
[0659] The results are shown in FIG. 8. The genes that belonged to
the MAPK pathway and the p53 pathway were determined based on
pathway identities from KEGG.
[0660] The results indicate that STK11 copy number loss is
associated with or predictive of nonresponsiveness or resistance
(e.g., acquired resistance) of a cancer (e.g., a CLL) to a PI3K
inhibitor (e.g., Compound 1).
[0661] In addition, the results indicate that a dual pathway
alteration (a mutation in both MAPK and p53 pathways) is associated
with or predictive of nonresponsiveness or resistance (e.g.,
acquired resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor
(e.g., Compound 1). Genes in the MAPK and p53 pathways that were
frequently mutated are indicated in Tables 18 and 20 below.
[0662] Furthermore, the results indicate that copy number loss of
STK11 combined with copy number loss of TSC1, TSC2, or both (shown
as "STK11/TSC loss" in FIG. 8) is associated with or predictive of
nonresponsiveness or resistance (e.g., acquired resistance) of a
cancer (e.g., a CLL) to a PI3K inhibitor (e.g., Compound 1).
[0663] Mutations in TP53 were further characterized, by determining
the frequency of TP35 mutations in responders versus
non-responders. Specifically, Table 15 below shows that TP53
alterations, including loss of TP53 and TP53 mutations, were more
common in non-responders than responders. Thus, loss of TP53
correlated with a poorer prognosis.
TABLE-US-00015 TABLE 15 CR/PR SD/PD P value Genetic alterations (n
= 32) (n = 23) (Fisher's exact) Loss of TP53 6 11 0.0368* TP53
mutation 6 7 0.34 Both 3 5 0.2573 Any TP53 alterations 9 13
0.0511
[0664] FIGS. 9 and 10 show the results of a re-analysis of the same
data that were used in the analysis presented in Example 9, except
that PR patients with lymphocytosis were classified as
non-responders, whereas such patients were classified as responders
in Example 9.
[0665] The results of the re-analysis confirmed that STK11 copy
number loss is associated with or predictive of nonresponsiveness
or resistance (e.g., acquired resistance) of a cancer (e.g., a CLL)
to a PI3K inhibitor (e.g., Compound 1). Furthermore, the results
confirmed that a dual pathway alteration (a mutation in both MAPK
and p53 pathways) and mutation of BCR pathway is associated with or
predictive of nonresponsiveness or resistance (e.g., acquired
resistance) of a cancer (e.g., a CLL) to a PI3K inhibitor (e.g.,
Compound 1).
[0666] FIG. 11 shows additional results of an analysis of
relationships between mutations and copy number variations and
responses. Correlations between CLL common CNVs and response to
Compound 1 are shown in FIG. 12.
Example 11
Additional Data Regarding CNVs and Mutations in CLL Patients
[0667] Using the methods described in the Examples above, CNVs that
are more frequently present in non-responders versus responders to
Compound 1 were determined. The results are shown in Table 16.
TABLE-US-00016 TABLE 16 CNVs that are more frequently present in
Compound 1 non-responders. Chromosome Region location Gene Event
chr19: 1,205,798-1,228,434 19p13.3 STK11 Copy number loss chr9:
135,766,735-135,820,020 9q34.13 TSC1 Copy number loss chr16:
2,097,990-2,138,713 16p13.3 TSC2 Copy number loss
[0668] In total 140 genes were detected with baseline mutations in
Compound 1-treated CLL patients (Table 17).
TABLE-US-00017 TABLE 17 List of genes that have mutations detected
in Compound 1-treated CLL patients GeneName Refseq ID ABCA13
NM_152701 ABCA7 NM_019112 ADAMTSL3 NM_207517 AKAP8 NM_005858 ALK
NM_004304 ARID1A NM_006015 ARID1B NM_020732 ASXL1 NM_015338 ATM
NM_000051 ATR NM_001184 ATRX NM_000489 Bcl11A NM_022893 Bcl2
NM_000633 BCR NM_004327 BIRC3 NM_001165 BRAF NM_004333 BTG1
NM_001731 BTK NM_001287344 CARD11 NM_032415 CBFA2T3 NM_005187 CBL
NM_005188 CCND3 NM_001287427 CCT6B NM_006584 CD36 NM_001001548
CDC73 NM_024529 CDH1 NM_004360 CDH11 NM_001797 CIC NM_015125 CIITA
NM_001286402 COL4A2 NM_001846 CREBBP NM_004380 CSMD1 NM_033225
CSMD3 NM_198123 DAXX NM_001141969 DCHS1 NM_003737 DEK NM_003472
DIS3 NM_014953 DNM2 NM_001005361 DNMT1 NM_001130823 DPYD NM_000110
DST NM_001144769 EP300 NM_001429 EPHB1 NM_004441 EPHB2 NM_004442
ERBB4 NM_005235 ETV6 NM_001987 FAT2 NM_001447 FAT4 NM_024582 FBXO11
NM_001190274 FBXW7 NM_033632 FGFR1 NM_001174064 FGFR2 NM_022970
FGFR4 NM_213647 FLT3 NM_004119 FOXO1 NM_002015 FTCD NM_006657 FUBP1
NM_003902 FUS NM_004960 GNAQ NM_002072 GNAS NM_001077490 GRM8
NM_001127323 H3F3A NM_002107 HLF NM_002126 HNF1A NM_000545 HOXC13
NM_017410 HRAS NM_176795 IDH1 NM_001282387 JAK3 NM_000215 KIT
NM_000222 LPHN3 NM_015236 LRP1B NM_018557 LRRK2 NM_198578 MAF
NM_001031804 MAGI1 NM_015520 MALT1 NM_006785 MAP2K1 NM_002755
MAP3K1 NM_005921 MDM2 NM_002392 MED12 NM_005120 MEF2B NM_001145785
MKL1 NM_001282662 MSH2 NM_000251 MSH6 NM_000179 MTOR NM_004958 MYC
NM_002467 MYD88 NM_001172567 NCOA2 NM_006540 NCOR1 NM_006311 NF1
NM_001042492 NIN NM_020921 NKX2-1 NM_003317 NOTCH1 NM_017617 NOTCH2
NM_024408 NSD1 NM_022455 NTRK1 NM_002529 NTRK3 NM_001007156 NUMA1
NM_001286561 NUP214 NM_005085 NUP98 NM_016320 OGT NM_181672 PCDH15
NM_001142771 POLO NM_033026 PCM1 NM_006197 PCSK7 NM_004716 PDE4DIP
NM_014644 PDGFRA NM_006206 PDGFRB NM_002609 PERI NM_002616 PKHD1
NM_138694 PLCG2 NM_002661 PML NM_033238 PMS2 NM_000535 PRDM16
NM_022114 PRKDC NM_006904 PTCH1 NM_001083602 PTPRD NM_002839 PTPRT
NM_133170 RALGDS NM_006266 RB1 NM_000321 RELN NM_005045 RNF213
NM_001256071 ROBO2 NM_001290040 RYR1 NM_000540 SETD2 NM_014159
SF3B1 NM_012433 SH2B3 NM_005475 SMARCA4 NM_001128844 STAT6
NM_001178078 SUZ12 NM_015355 SYNE1 NM_182961 TALI NM_003189 TCF3
NM_003200 TET1 NM_030625 TET2 NM_001127208 TLL2 NM_012465 TNFAIP3
NM_001270508 TP53 NM_001276696 TRIP11 NM_004239 XPO1 NM_003400
ZRSR2 NM_005089
[0669] Frequently altered signaling pathways in Compound 1
non-responders and the involved genes and mutation sites are shown
in Table 18, Table 29 and Table 20.
[0670] In the MAPK and ERBB signaling pathways, 16 genes were
frequently mutated: BIRC3, BRAF, CBL, ERBB4, FGFR1, FGFR2, FGFR4,
FLT3, HRAS, MAP2K1, MAP3K1, MTOR, MYC, NF1, NTRK1, PDGFRA and
PDGFRB. See Table 18.
[0671] In the BCR pathway, 7 genes were frequently mutated: Bcl2,
BTK, CARD11, MALT1, MTOR, MYD88 AND PLCG2. See Table 19.
[0672] In the p53 signaling and cell cycle pathways 12 genes were
frequently mutated: ATM, ATR, CCND3, MYC, CREBBP, EP300, FBXW7,
MDM2, PRKDC, RB1, TP53 and XPO1. See Table 20.
[0673] In addition, mutations in the JAK/STAT, NF-kB, and apoptosis
pathways are enriched in IWCLL non-responders.
[0674] FIG. 14 shows relationships between response and alterations
in genes of various pathways, including the MAPK pathway p53
pathway, dual p53 and MAPK pathways, and BCR pathway.
TABLE-US-00018 TABLE 18 The frequently mutated MAPK pathway genes
and mutation sites in Compound 1 non-responders. Reference Mutation
Pathway GeneName Refseq ID Chromosome Position Allele Allele
AAMutation MAPK BIRC3 NM_001165 11 102201966 G G-AATC E440DEL BRAF
NM_004333 7 140534536 G C S126C CBL NM_005188 11 119155775 C G
P510A ERBB4 NM_005235 2 212295820 C A M831I ERBB4 NM_005235 2
212989562 C T R50H ERBB4 NM_005235 2 213403221 T A S12C FGFR1
NM_001174064 8 38272320 C T D642N FGFR2 NM_022970 10 123310807 G C
Y207* FGFR4 NM_213647 5 176520277 A C H399P FGFR4 NM_213647 5
176517445 T G L49R FLT3 NM_004119 13 28623641 T G N306H HRAS
NM_176795 11 533509 C T D132N MAP2K1 NM_002755 15 66727455 G T K57N
MAP2K1 NM_002755 15 66782068 C G N345K MAP3K1 NM_005921 5 56168815
G T A557S MTOR NM_004958 1 11204742 C T R1612Q MTOR NM_004958 1
11301623 C T A510T MYC NM_002467 8 128750680 A C T73P NF1
NM_001042492 17 29557906 A C N1054H NTRK1 NM_002529 1 156836766 G A
E142K PDGFRA NM_006206 4 55139810 G A A491T PDGFRA NM_006206 4
5512932 G A E156K PDGFRB NM_002609 5 149510109 G A L454F
TABLE-US-00019 TABLE 19 The frequently mutated BCR pathway genes
and mutation sites in Compound 1 non-responders. Reference Mutation
Pathway GeneName Refseq ID Chromosome Position Allele Allele
AAMutation BCR Bel2 NM_000633 18 60985508 G T A131D BTK
NM_001287344 X 100611164 C A C481F BTK NM-001287344 X 100611164 C G
C481S CARD11 NM_032415 7 2959106 G A R804C MALT1 NM_006785 18
56411677 A G K621E MALT1 NM_006785 18 56414750 G A M717I MTOR
NM_004958 1 11204742 C T R1612Q MTOR NM_004958 1 11301623 C T A510T
MYC NM_002467 8 128750680 A C T73P MYD88 NM_001172567 3 38182025 G
T V217F MYD88 NM_001172567 3 38182337 C T P266L PLCG2 NM_02661 16
81973605 T G M1141R PLCG2 NM_002661 16 81953154 C T S707F
TABLE-US-00020 TABLE 20 The frequently mutated p53/cell cycle
pathway genes and mutation sites in Compound 1 non-responders.
Reference Mutation Pathway Gene Name Refseq ID Chromosome Position
Allele Allele AAMutation p53/Cell ATM NM_000051 11 108196836 G A
G2287R cycle ATM NM_000051 11 108129788 A A-TTTGTAAAAG I818DEL ATM
NM_000051 11 108200967 T A L2445Q ATM NM_000051 11 108164152 G T
R1575L ATM NM_000051 11 108186596 T C L2018S ATM NM_000051 111
108115724 A G H291R ATR NM_001184 3 142274725 T A K779* ATR
NM_001184 3 142274853 C A G736V CCND3 NM_001287427 6 41904413 G A
P149S CCND3 NM_001287427 6 41903707 G A P234S CREBBP NM_004380 16
3795324 T A M1290L EP300 NM_001429 22 41574510 T T-CAG L2265DEL
EP300 NM_001429 22 41513811 C G P239A FBXW7 NM_033632 4 153253763 T
A K324* MDM2 NM_002392 12 69233526 T G L464R MDM2 NM_002392 12
69233160 A G K342R MDM2 NM_002392 12 69233130 G A R332H MDM2
NM_002392 12 69233252 G A V373M PRKDC NM_006904 8 48855869 T C
N289S PRKDC NM_006904 8 48761821 C G V2391L PRKDC NM_006904 8
48691647 T C R3832G PRKDC NM_006904 8 48767904 G A R2213* RB1
NM_000321 13 48878084 C C-GCCGCCGCT T12DEL RB1 NM_000321 13
49039396 G C S794T TP53 NM_001276696 17 7578199 A C V178G TP53
NM_001276696 17 7578196 A T V179E TP53 NM_001276696 17 7578211 C A
R174L TP53 NM_001276696 17 7578437 G A Q126* TP53 NM_001276696 17
7578508 C T C102Y TP53 NM_001276696 17 7577114 C T C236Y TP53
NM_001276696 17 7578221 T T-TC R209DEL TP53 NM_001276696 17 7578394
G C H140R TP53 NM_001276696 17 7578272 G A H154Y TP53 NM_001276696
17 7578554 A C Y87D TP53 NM_001276696 17 7578394 T C H140R TP53
NM_001276696 17 7578484 G G-A S110DEL TP53 NM_001276696 17 7578263
G C R157G TP53 NM_001276696 17 7577144 A G L226P TP53 NM_001276696
17 7577123 A T V233E XPO1 NM_003400 2 61719472 C T E571K
Example 12
PTEN is a Biomarker for Compound 1 Resistance
[0675] Experiments were performed to assess the expression of PTEN
in cells that were resistant to Compound 1. Compound 1 resistant
cells were generated by culturing cells in the presence of Compound
1 or DMSO as a control for 8 weeks. Cells were subcloned under
selective pressure from the drug, seeding at densities of 3 cells
per well, 1 cell per well, or 0.3 cell per well. Parental,
DMSO-treated, and Compound 1-resistant (also referred to as
Compound 1-R) clones were selected for expansion. Five clones from
each group were expanded. Cells were harvested for various assays,
including CTG (CellTiter-Glo, Promega, an assay that measures ATP
levels as a surrogate for cell number in order to observe cell
viability and changes in proliferation rate), PD (pharmacodynamic),
RNA analysis, DNA analysis, and short tandem repeat (STR)
fingerprinting. A CTG assay was performed to confirm that cells
were resistant to Compound 1 at the time of sample collection. As
shown in FIG. 15 and Table 21, the average 1050 for Compound 1
inhibition of the resistant cells was higher than the control
cells. RNA-seq experiments were also performed on the samples from
DMSO control and Compound 1 resistant cells. Five clones of
each--DMSO-treated control cells that are not resistant to Compound
1, and Compound 1 resistant cells--were tested. As shown in FIG.
16, there was a substantial downregulation in PTEN expression in
Compound 1 resistant cell clones, but not in the DMSO
control-treated cell clones. This downregulation in PTEN expression
was seen at the RNA level as well as the protein level. See, e.g.,
FIG. 16. These results show that PTEN is a biomarker for Compound 1
resistance, where low PTEN levels correlate with resistance.
TABLE-US-00021 TABLE 21 AVG Compound 1 Clones IC50 (nM) Control 241
.+-. 17 Compound 1 resistant 5420 .+-. 1079
EQUIVALENTS
[0676] While this invention has been disclosed with reference to
specific aspects, it is apparent that other aspects and variations
of this invention can be devised by others skilled in the art
without departing from the true spirit and scope of the invention.
The appended claims are intended to be construed to include all
such aspects and equivalent variations.
Sequence CWU 1
1
2111DNAHomo sapiens 1atttgtaaaa g 11210DNAHomo sapiens 2cgccgccgct
10
* * * * *