U.S. patent application number 13/230315 was filed with the patent office on 2012-09-13 for combination cancer therapy with bis(thiohydrazide) amide compounds.
This patent application is currently assigned to Synta Pharmaceuticals Corp.. Invention is credited to Thomas A. Dahl, Matthew McLeod.
Application Number | 20120232134 13/230315 |
Document ID | / |
Family ID | 36694110 |
Filed Date | 2012-09-13 |
United States Patent
Application |
20120232134 |
Kind Code |
A1 |
Dahl; Thomas A. ; et
al. |
September 13, 2012 |
COMBINATION CANCER THERAPY WITH BIS(THIOHYDRAZIDE) AMIDE
COMPOUNDS
Abstract
Disclosed is a method of treating a subject with cancer, the
method comprising the step of co-administering to the subject over
three to five weeks, a taxane in an amount of between about 243
.mu.mol/m.sup.2 to 315 .mu.mol/m.sup.2; and a bis(thiohydrazide
amide) compound in an amount between about 1473 .mu.mol/m.sup.2 and
about 1722 .mu.mol/m.sup.2.
Inventors: |
Dahl; Thomas A.; (Cambridge,
MA) ; McLeod; Matthew; (Boston, MA) |
Assignee: |
Synta Pharmaceuticals Corp.
Lexington
MA
|
Family ID: |
36694110 |
Appl. No.: |
13/230315 |
Filed: |
September 12, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11918357 |
Aug 25, 2008 |
8017654 |
|
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PCT/US06/14531 |
Apr 13, 2006 |
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13230315 |
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60672139 |
Apr 15, 2005 |
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Current U.S.
Class: |
514/449 |
Current CPC
Class: |
A61K 31/16 20130101;
A61K 31/337 20130101; A61K 45/06 20130101; A61P 35/00 20180101;
A61P 35/04 20180101; A61K 31/16 20130101; A61K 31/337 20130101;
A61K 31/165 20130101; A61K 31/165 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/449 |
International
Class: |
A61K 31/337 20060101
A61K031/337; A61P 35/00 20060101 A61P035/00 |
Claims
1. A method of treating a human with ovarian cancer comprising the
step of co-administering to the human over three to five weeks:
paclitaxel or a paclitaxel analogue in an amount of between about
243 .mu.mol/m.sup.2 to 315 .mu.mol/m.sup.2; and a bis(thiohydrazide
amide) in an amount between about 1473 .mu.mol/m.sup.2 and about
1722 .mu.mol/m.sup.2, wherein the bis(thiohydrazide amide) is
represented by the following Structural Formula: ##STR00022## or a
pharmaceutically acceptable salt thereof.
2. The method of claim 1, wherein the bis(thiohydrazide amide) is
administered in combination with paclitaxel or docetaxel.
3. The method of claim 2, wherein the bis(thiohydrazide amide) is
administered in combination with paclitaxel.
4. The method of claim 3, wherein the paclitaxel and the
bis(thiohydrazide amide) are each administered in three equal
weekly doses of three weeks of a four week period.
5. The method of claim 4, further comprising repeating the four
week administration period until the cancer is in remission.
6. The method of claim 5, wherein the paclitaxel is intravenously
administered in a weekly dose of about 94 .mu.mol/m.sup.2.
7. The method of claim 6, wherein the bis(thiohydrazide amide) is
intravenously administered in a weekly dose of between about 500
.mu.mol/m.sup.2 and about 562 .mu.mol/m.sup.2.
8. The method of claim 7, wherein the bis(thiohydrazide amide) is
intravenously administered in a weekly dose of 532
.mu.mol/m.sup.2.
9. The method of claim 1, wherein the bis(thiohydrazide amide) is
the disodium or dipotassium salt.
10. A method of treating a human with cancer, comprising
intravenously administering to the human in a four week period,
three equal weekly doses of the paclitaxel in an amount of about 94
.mu.mol/m.sup.2 and a bis(thiohydrazide amide) represented by the
following Structural Formula: ##STR00023## or a pharmaceutically
acceptable salt thereof in an amout of about 532 .mu.mol/m.sup.2,
wherein the cancer is ovarian cancer.
Description
RELATED APPLICATION
[0001] This application is a continuation of the U.S. application
Ser. No. 11/918,357, filed on Aug. 25, 2008, issuing, which is a
U.S. National Stage of International Application No.
PCT/US/2006/014531, filed Apr. 13, 2006, which claims the benefit
of U.S. Provisional Application No. 60/672,139, filed on Apr. 15,
2005. The entire teachings of the above applications are
incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] The taxanes are an important class of anticancer agents. In
particular, Taxol.TM. (paclitaxel) is an effective anticancer
agent, especially in the treatment of ovarian cancer, metastatic
breast cancer, non-small cell lung cancer (NSCLC) and AIDS-related
Kaposi's sarcoma. However, there is still a significant need in the
art for improvement in the efficacy of paclitaxel therapy, both in
terms of the proportion of patients who respond to therapy and the
survival benefit imparted. Moreover, administration of Taxol has
side effects, including reducing immune function by reducing
natural killer (NK) cell activity.
[0003] In an attempt to improve efficacy, paclitaxel is sometimes
used in combination with other anticancer agents. For example,
carboplatin in the treatment of NSCLC. Such combinations can have
an additive benefit or increased response rate, but can tend to
also combine the side effect profiles of each agent. Other agents
have been researched, for example, bis(thiohydrazide amides) have
been tested in animal models as described in U.S. Pat. Nos.
6,800,660, 6,762,204, U.S. patent application Ser. Nos. 10/345,885
filed Jan. 15, 2003, and 10/758,589, Jan. 15, 2004, the entire
teachings of which are incorporated herein by reference.
[0004] However, there is still an urgent need for particular
combination therapies that can enhance the antitumor effects of
paclitaxel without further increasing side effects suffered by
patients.
SUMMARY OF THE INVENTION
[0005] It is now found that certain bis(thiohydrazide) amide and
taxane combinations are surprisingly effective at treating subjects
with cancer without further increasing side effects. The particular
combination therapies disclosed herein demonstrate surprising
biological activity by raising Hsp70 levels (see Example 3), by
demonstrating significant anticancer effects (see Examples 4-5),
and by halting or reversing side effects (see Examples 4-5) such as
the reduction in natural killer (NK) cell activity typically
associated with Taxol.TM. administration.
[0006] A method of treating a subject with cancer includes the step
of co-administering to the subject over three to five weeks, a
taxane in an amount of between about 243 .mu.mol/m2 to 315
.mu.mol/m2 (e.g., equivalent to paclitaxel in about 210-270 mg/m2);
and a bis(thiohydrazide amide) in an amount between about 1473
.mu.mol/m2 and about 1722 .mu.mol/m2 (e.g., Compound (1) in about
590-690 mg/m2). The bis(thiohydrazide amide) is represented by
Structural Formula I:
##STR00001##
[0007] Y is a covalent bond or an optionally substituted straight
chained hydrocarbyl group, or, Y, taken together with both
>C.dbd.Z groups to which it is bonded, is an optionally
substituted aromatic group.
[0008] R.sub.1-R.sub.4 are independently --H, an optionally
substituted aliphatic group, an optionally substituted aryl group,
or R.sub.1 and R.sub.3 taken together with the carbon and nitrogen
atoms to which they are bonded, and/or R.sub.2 and R.sub.4 taken
together with the carbon and nitrogen atoms to which they are
bonded, form a non-aromatic heterocyclic ring optionally fused to
an aromatic ring.
[0009] R.sub.7-R.sub.8 are independently --H, an optionally
substituted aliphatic group, or an optionally substituted aryl
group.
[0010] Z is O or S.
[0011] In various embodiments, a method of treating a subject with
cancer includes administering to the subject effective amounts of
each of a platinum anticancer compound; a taxane or a
pharmaceutically acceptable salt or solvate thereof; and a
bis(thiohydrazide amide) represented by Structural Formula I or a
pharmaceutically acceptable salt or solvate thereof.
[0012] In various embodiments, a method of treating a subject with
cancer includes administering to the subject once every three
weeks, independently or together a taxane in an amount of about 205
.mu.mol/m2 (e.g., paclitaxel in about 175 mg/m2); and a
bis(thiohydrazide amide) represented by Structural Formula I or a
pharmaceutically acceptable salt or solvate thereof in an amount
between about 220 .mu.mol/m2 and about 1310 .mu.mol/m2 (e.g.,
Compound (1) in about 88-525 mg/m2).
[0013] In various embodiments, a pharmaceutical composition
includes a pharmaceutically acceptable carrier or diluent. In some
embodiments, the molar ratio of bis(thiohydrazide amide) to taxane
can be between about 5.5:1 and about 5.9:1, in certain embodiments,
between about 2.7:1 and about 2.9:1, and in particular embodiments,
between about 4.1:1 and about 4.5:1.
[0014] In various embodiments, the invention includes the use of a
bis(thiohydrazide amide) for the manufacture of medicament for
treating cancer in combination with a taxane in each of the molar
ratios described above. In some embodiments, the invention includes
the use of a bis(thiohydrazide amide) and taxane for the
manufacture of medicament for treating cancer in each of the molar
ratios described above.
[0015] The taxanes employed in the invention, e.g., paclitaxel, are
described in the Detailed Description section below.
[0016] In various embodiments, a pharmaceutically acceptable salt
or solvate of either the bis(thiohydrazide)amide or taxane
anticancer agents can be employed, optionally with a
pharmaceutically acceptable carrier or diluent. In certain
embodiments, a pharmaceutical composition includes the
bis(thiohydrazide) amide, the taxane, and a pharmaceutically
acceptable carrier or diluent.
[0017] The methods are particularly effective for treating the
claimed cancers as demonstrated in the Examples, and halting or
reversing side effects such as the reduction in natural killer (NK)
cell activity typically associated with Taxol.TM.
administration.
BRIEF DESCRIPTION OF THE DRAWINGS
[0018] FIGS. 1A, 1B, and 1C are bar graphs showing the percent
increase in Hsp70 plasma levels associated with administration of
the Compound (1)/paclitaxel combination therapy at 1 hour (FIG.
1A), 5 hours (FIG. 1B), and 8 hours (FIG. 1C) after
administration.
DETAILED DESCRIPTION OF THE INVENTION
[0019] A description of preferred embodiments of the invention
follows.
[0020] In various embodiments, a method of treating a subject with
cancer includes the step of co-administering to the subject over
three to five weeks, a taxane in an amount of between about 243
.mu.mol/m2 to 315 .mu.mol/m2 (e.g., equivalent to paclitaxel in
about 210-270 mg/m2); and a bis(thiohydrazide amide) (e.g., as
represented by Structural Formula I) in an amount between about
1473 .mu.mol/m2 and about 1722 .mu.mol/m2 (e.g., Compound (1) in
about 590-690 mg/m2).
[0021] A subject, e.g., typically a human subject, can be treated
for any cancer described herein. Typically, the cancer can be a
soft tissue sarcoma (e.g., typically soft tissue sarcomas other
than GIST) or metastatic melanoma. In some embodiments, the cancer
is metastatic melanoma.
[0022] In some embodiments, the taxane and the bis(thio-hydrazide)
amide can each be administered in three equal weekly doses for
three weeks of a four week period. In preferred embodiments, the
four week administration period can be repeated until the cancer is
in remission.
[0023] The taxane can be any taxane defined herein. In particular
embodiments, the taxane is paclitaxel intravenously administered in
a weekly dose of about 94 .mu.mol/m2 (80 mg/m2).
[0024] In various embodiments, the bis(thiohydrazide amide) can be
intravenously administered in a weekly dose of between about 500
.mu.mol/m2 and about 562 .mu.mol/m2, or more typically in a weekly
dose of about 532 .mu.mol/m2. (e.g., Compound (1) in about 590-690
mg/m2).
[0025] In some embodiments, the subject is treated for metastatic
melanoma. In certain embodiments, the subject is treated for soft
tissue sarcomas other than GIST.
[0026] In preferred embodiments, a method of treating a human
subject with cancer includes intravenously administering to the
subject in a four week period, three equal weekly doses of
paclitaxel in an amount of about 94 .mu.mol/m2; and a
bis(thiohydrazide amide) represented by the following Structural
Formula:
##STR00002##
or a pharmaceutically acceptable salt or solvate thereof in an
amount of about 532 .mu.mol/m2. Typically, the cancer is a soft
tissue sarcomas (e.g., typically soft tissue sarcomas other than
GIST) or metastatic melanoma.
[0027] In various embodiments, the subject can be intravenously
administered between about 220 .mu.mol/m2 and about 1310 .mu.mol/m2
(e.g., Compound (1) in about 88-525 mg/m2) of the bis(thiohydrazide
amide) once every 3 weeks, generally between about 220 .mu.mol/m2
and about 1093 .mu.mol/m2 (e.g., Compound (1) in about 88-438
mg/m2) once every 3 weeks, typically between about 624 .mu.mol/m2
and about 1124 .mu.mol/m2 m2 (e.g., Compound (1) in about 250-450
mg/m2), more typically between about 811 .mu.mol/m2 and about 936
.mu.mol/m2 m2 (e.g., Compound (1) in about 325-375 mg/m2), or in
particular embodiments, about 874 .mu.mol/m2 ((e.g., Compound (1)
in about 350 mg/m2). In particular embodiments, the subject can be
intravenously administered between about 582 .mu.mol/m2 and about
664 .mu.mol/m2 (e.g., Compound (1) in about 233-266 mg/m2) of the
bis(thiohydrazide amide) once every 3 weeks. In certain
embodiments, the bis(thiohydrazide amide) is in an amount of about
664 .mu.mol/m2 (e.g., Compound (1) in about 266 mg/m2).
[0028] In various embodiments, the subject can be intravenously
administered between about 200 .mu.mol/m2 to about 263 .mu.mol/m2
of the taxane as paclitaxel once every 3 weeks (e.g., paclitaxel in
about 175-225 mg/m2). In some embodiments, the subject can be
intravenously administered between about 200 .mu.mol/m2 to about
234 .mu.mol/m2 of the taxane as paclitaxel once every 3 weeks
(e.g., paclitaxel in about 175-200 mg/m2). In certain embodiments,
the paclitaxel is administered in an amount of about 234 .mu.mol/m2
(200 mg/m2). In certain embodiments, the paclitaxel is administered
in an amount of about 205 .mu.mol/m2 (175 mg/m2)
[0029] In various embodiments, the taxane, e.g., paclitaxel, and
the bis(thiohydrazide amide), e.g., Compound (1), can be
administered together in a single pharmaceutical composition.
[0030] In various embodiments, a method of treating a subject with
cancer includes administering to the subject once every three
weeks, independently or together a taxane in an amount of about 205
.mu.mol/m2 (e.g., paclitaxel in about 175 mg/m2); and a
bis(thiohydrazide amide) represented by Structural Formula I or a
pharmaceutically acceptable salt or solvate thereof in an amount
between about 220 .mu.mol/m2 and about 1310 .mu.mol/m2 (e.g.,
Compound (1) in about 88-525 mg/m2). Typically, the taxane is
paclitaxel intravenously administered in an amount of about 205
.mu.mol/m2. The bis(thiohydrazide amide) can typically be
intravenously administered between about 220 .mu.mol/m2 and about
1093 .mu.mol/m2 (e.g., Compound (1) in about 88-438 mg/m2), more
typically between about 749 .mu.mol/m2 and about 999 .mu.mol/m2
(e.g., compound (1) in about 300-400 mg/m2), in some embodiments
between about 811 .mu.mol/m2 and about 936 .mu.mol/m2 (e.g.,
Compound (1) in about 325-375 mg/m2). In certain embodiments, the
bis(thiohydrazide amide) can be Compound (1) intravenously
administered between about 874 .mu.mol/m2 (about 350 mg/m2).
[0031] In a particular embodiment, a method of treating a subject
with cancer includes intravenously administering to the subject in
a single dose per three week period: paclitaxel in an amount of
about 205 .mu.mol/m2 (175 mg/m2); and Compound (1) or a
pharmaceutically acceptable salt or solvate thereof in an amount of
about 874 .mu.mol/m2 (350 mg/m2), wherein the cancer is a soft
tissue sarcomas other than GIST or metastatic melanoma.
[0032] In various embodiments, a pharmaceutical composition
includes a pharmaceutically acceptable carrier or diluent; and a
molar ratio of a bis(thiohydrazide amide) to a taxane between about
5.5:1 and about 5.9:1, wherein the bis(thiohydrazide amide)
represented by Structural Formula I or a pharmaceutically
acceptable salt or solvate thereof. In some embodiments, the molar
ratio of the bis(thiohydrazide amide) to the taxane is between
about 5.6:1 and about 5.8:1, or more typically, about 5.7:1. In
certain embodiments, the taxane is paclitaxel or a pharmaceutically
acceptable salt or solvate thereof. In particular embodiments, the
bis(thiohydrazide amide) is Compound (1).
[0033] In various embodiments, a pharmaceutical composition
includes a pharmaceutically acceptable carrier or diluent; and a
molar ratio of a bis(thiohydrazide amide) to a taxane between about
2.6:1 and about 3.0:1, wherein the bis(thiohydrazide amide)
represented by Structural Formula I or a pharmaceutically
acceptable salt or solvate thereof. In some embodiments, the molar
ratio of the bis(thiohydrazide amide) to the taxane is between
about 2.7:1 and about 2.9:1, or more typically, about 2.8:1. In
certain embodiments, the taxane is paclitaxel or a pharmaceutically
acceptable salt or solvate thereof. In particular embodiments, the
bis(thiohydrazide amide) is Compound (1).
[0034] In various embodiments, a pharmaceutical composition
includes a pharmaceutically acceptable carrier or diluent; and a
molar ratio of a bis(thiohydrazide amide) to a taxane between about
4.1:1 and about 4.5:1, wherein the bis(thiohydrazide amide)
represented by Structural Formula I or a pharmaceutically
acceptable salt or solvate thereof. In some embodiments, the molar
ratio of the bis(thiohydrazide amide) to the taxane is between
about 4.2:1 and about 4.4:1, or more typically, about 4.3:1. In
certain embodiments, the taxane is paclitaxel or a pharmaceutically
acceptable salt or solvate thereof. In particular embodiments, the
bis(thiohydrazide amide) is Compound (1).
[0035] In various embodiments, the invention includes the use of a
bis(thiohydrazide amide) for the manufacture of medicament for
treating cancer in combination with a taxane in a molar ratio of
bis(thiohydrazide amide) to taxane between about 5.5:1 and about
5.9:1, typically between about 5.6:1 and about 5.8:1, more
typically about 5.7:1, wherein the bis(thiohydrazide amide) is
represented by Structural Formula I. In some embodiments, the molar
ratio of bis(thiohydrazide amide) to taxane can be between about
2.6:1 and about 3.0:1, typically between about 2.7:1 and about
2.9:1, more typically about 2.8:1. In some embodiments, the molar
ratio of bis(thiohydrazide amide) to taxane can be between about
4.1:1 and about 4.5:1, typically between about 4.2:1 and about
4.4:1, more typically about 4.3:1.
[0036] In various embodiments, the invention includes the use of a
bis(thiohydrazide amide) and taxane for the manufacture of
medicament for treating cancer in a molar ratio of
bis(thiohydrazide amide) to taxane between about 5.5:1 and about
5.9:1, typically between about 5.6:1 and about 5.8:1, more
typically about 5.7:1, wherein the bis(thiohydrazide amide) is
represented by Structural Formula I. In some embodiments, the molar
ratio of bis(thiohydrazide amide) to taxane can be between about
2.6:1 and about 3.0:1, typically between about 2.7:1 and about
2.9:1, more typically about 2.8:1. In some embodiments, the molar
ratio of bis(thiohydrazide amide) to taxane can be between about
4.1:1 and about 4.5:1, typically between about 4.2:1 and about
4.4:1, more typically about 4.3:1.
[0037] The bis(thiohydrazide amides) employed in the disclosed
invention are represented by Structural Formula I, or a
pharmaceutically acceptable salt or solvate thereof.
[0038] In one embodiment, Y in Structural Formula I is a covalent
bond, --C(R.sub.5R.sub.6)--, --(CH.sub.2CH.sub.2)--,
trans-(CH.dbd.CH)--, cis-(CH.dbd.CH)-- or --(C.ident.C)-- group,
preferably --C(R.sub.5R.sub.6)--. R.sub.1-R.sub.4 are as described
above for Structural Formula I. R.sub.5 and R.sub.6 are each
independently --H, an aliphatic or substituted aliphatic group, or
R.sub.5 is --H and R.sub.6 is an optionally substituted aryl group,
or, R.sub.5 and R.sub.6, taken together, are an optionally
substituted C2-C6 alkylene group. The pharmaceutically acceptable
cation is as described in detail below.
[0039] In specific embodiments, Y taken together with both
>C.dbd.Z groups to which it is bonded, is an optionally
substituted aromatic group. In this instance, certain
bis(thiohydrazide amides) are represented by Structural Formula
II:
##STR00003##
wherein Ring A is substituted or unsubstituted and V is --CH-- or
--N--. The other variables in Structural Formula II are as
described herein for Structural Formula I or III.
[0040] In particular embodiments, the bis(thiohydrazide amides) are
represented by Structural Formula III:
##STR00004##
[0041] R.sub.1-R.sub.8 and the pharmaceutically acceptable cation
are as described above for Structural Formula I.
[0042] In Structural Formulas I-III, R.sub.1 and R.sub.2 are the
same or different and/or R.sub.3 and R.sub.4 are the same or
different; preferably, R.sub.1 and R.sub.2 are the same and R.sub.3
and R.sub.4 are the same. In Structural Formulas I and III, Z is
preferably O. Typically in Structural Formulas I and III, Z is O;
R.sub.1 and R.sub.2 are the same; and R.sub.3 and R.sub.4 are the
same. More preferably, Z is O; R.sub.1 and R.sub.2 are the same;
R.sub.3 and R.sub.4 are the same, and R.sub.7 and R.sub.8 are the
same.
[0043] In other embodiments, the bis(thiohydrazide amides) are
represented by Structural Formula III: R.sub.1 and R.sub.2 are each
an optionally substituted aryl group, preferably an optionally
substituted phenyl group; R.sub.3 and R.sub.4 are each an
optionally substituted aliphatic group, preferably an alkyl group,
more preferably, methyl or ethyl; and R.sub.5 and R.sub.6 are as
described above, but R.sub.5 is preferably --H and R.sub.6 is
preferably --H, an aliphatic or substituted aliphatic group.
[0044] Alternatively, R.sub.1 and R.sub.2 are each an optionally
substituted aryl group; R.sub.3 and R.sub.4 are each an optionally
substituted aliphatic group; R.sub.5 is --H; and R.sub.6 is --H, an
aliphatic or substituted aliphatic group. Preferably, R.sub.1 and
R.sub.2 are each an optionally substituted aryl group; R.sub.3 and
R.sub.4 are each an alkyl group; and R.sub.5 is --H and R.sub.6 is
--H or methyl. Even more preferably, R.sub.1 and R.sub.2 are each
an optionally substituted phenyl group; R.sub.3 and R.sub.4 are
each methyl or ethyl; and R.sub.5 is --H and R.sub.6 is --H or
methyl. Suitable substituents for an aryl group represented by
R.sub.1 and R.sub.2 and an aliphatic group represented by R.sub.3,
R.sub.4 and R.sub.6 are as described below for aryl and aliphatic
groups.
[0045] In another embodiment, the bis(thiohydrazide amides) are
represented by Structural Formula III: R.sub.1 and R.sub.2 are each
an optionally substituted aliphatic group, preferably a C3-C8
cycloalkyl group optionally substituted with at least one alkyl
group, more preferably cyclopropyl or 1-methylcyclopropyl; R.sub.3
and R.sub.4 are as described above for Structural Formula I,
preferably both an optionally substituted alkyl group; and R.sub.5
and R.sub.6 are as described above, but R.sub.5 is preferably --H
and R.sub.6 is preferably --H, an aliphatic or substituted
aliphatic group, more preferably --H or methyl.
[0046] Alternatively, the bis(thiohydrazide amides) are represented
by Structural Formula III: R.sub.1 and R.sub.2 are each an
optionally substituted aliphatic group; R.sub.3 and R.sub.4 are as
described above for Structural Formula I, preferably both an
optionally substituted alkyl group; and R.sub.5 is --H and R.sub.6
is --H or an optionally substituted aliphatic group. Preferably,
R.sub.1 and R.sub.2 are both a C3-C8 cycloalkyl group optionally
substituted with at least one alkyl group; R.sub.3 and R.sub.4 are
both as described above for Structural Formula I, preferably an
alkyl group; and R.sub.5 is --H and R.sub.6 is --H or an aliphatic
or substituted aliphatic group. More preferably, R.sub.1 and
R.sub.2 are both a C3-C8 cycloalkyl group optionally substituted
with at least one alkyl group; R.sub.3 and R.sub.4 are both an
alkyl group; and R.sub.5 is --H and R.sub.6 is --H or methyl. Even
more preferably, R.sub.1 and R.sub.2 are both cyclopropyl or
1-methylcyclopropyl; R.sub.3 and R.sub.4 are both an alkyl group,
preferably methyl or ethyl; and R.sub.5 is --H and R.sub.6 is --H
or methyl.
[0047] In specific embodiments, the bis(thiohydrazide amides) are
represented by Structural Formula IV:
##STR00005##
[0048] wherein: R.sub.1 and R.sub.2 are both phenyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both phenyl, R.sub.3 and R.sub.4 are both
ethyl, and R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2
are both 4-cyanophenyl, R.sub.3 and R.sub.4 are both methyl,
R.sub.5 is methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
4-methoxyphenyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5
and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both phenyl,
R.sub.3 and R.sub.4 are both methyl, R.sub.5 is methyl, and R.sub.6
is --H; R.sub.1 and R.sub.2 are both phenyl, R.sub.3 and R.sub.4
are both ethyl, R.sub.5 is methyl, and R.sub.6 is --H; R.sub.1 and
R.sub.2 are both 4-cyanophenyl, R.sub.3 and R.sub.4 are both
methyl, and R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2
are both 2,5-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl,
and R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,5-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, R.sub.5
is methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
.sub.3-cyanophenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
.sub.3-fluorophenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
4-chlorophenyl, R.sub.3 and R.sub.4 are both methyl, R.sub.5 is
methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
2-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5
and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
3-methoxyphenyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5
and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,3-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,3-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, R.sub.5
is methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
2,5-difluorophenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,5-difluorophenyl, R.sub.3 and R.sub.4 are both methyl, R.sub.5 is
methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
2,5-dichlorophenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,5-dimethylphenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,5-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
phenyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5 and
R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
2,5-dimethoxyphenyl, R.sub.3 and R.sub.4 are both methyl, R.sub.5
is methyl, and R.sub.6 is --H; R.sub.1 and R.sub.2 are both
cyclopropyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5 and
R.sub.6 are both --H; R.sub.1 and R.sub.2 are both cyclopropyl,
R.sub.3 and R.sub.4 are both ethyl, and R.sub.5 and R.sub.6 are
both --H; R.sub.1 and R.sub.2 are both cyclopropyl, R.sub.3 and
R.sub.4 are both methyl, R.sub.5 is methyl, and R.sub.6 is --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, R.sub.5 is methyl and R.sub.6 is --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, R.sub.5 is ethyl, and R.sub.6 is --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, R.sub.5 is n-propyl, and R.sub.6 is --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both methyl;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 and
R.sub.4 are both ethyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both 1-methylcyclopropyl, R.sub.3 is
methyl, R.sub.4 is ethyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both 2-methylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both 2-phenylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both 1-phenylcyclopropyl, R.sub.3 and
R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are both cyclobutyl, R.sub.3 and R.sub.4 are
both methyl, and R.sub.5 and R.sub.6 are both --H; R.sub.1 and
R.sub.2 are both cyclopentyl, R.sub.3 and R.sub.4 are both methyl,
and R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
cyclohexyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5 and
R.sub.6 are both --H; R.sub.1 and R.sub.2 are both cyclohexyl,
R.sub.3 and R.sub.4 are both phenyl, and R.sub.5 and R.sub.6 are
both --H; R.sub.1 and R.sub.2 are both methyl, R.sub.3 and R.sub.4
are both methyl, and R.sub.5 and R.sub.6 are both --H; R.sub.1 and
R.sub.2 are both methyl, R.sub.3 and R.sub.4 are both t-butyl, and
R.sub.5 and R.sub.6 are both --H; R.sub.1 and R.sub.2 are both
methyl, R.sub.3 and R.sub.4 are both phenyl, and R.sub.5 and
R.sub.6 are both --H; R.sub.1 and R.sub.2 are both t-butyl, R.sub.3
and R.sub.4 are both methyl, and R.sub.5 and R.sub.6 are both --H;
R.sub.1 and R.sub.2 are ethyl, R.sub.3 and R.sub.4 are both methyl,
and R.sub.5 and R.sub.6 are both --H; or R.sub.1 and R.sub.2 are
both n-propyl, R.sub.3 and R.sub.4 are both methyl, and R.sub.5 and
R.sub.6 are both --H.
[0049] In specific embodiments, the bis(thiohydrazide amides) are
represented by Structural Formula V:
##STR00006##
wherein: R.sub.1 and R.sub.2 are both phenyl, and R.sub.3 and
R.sub.4 are both o-CH.sub.3-phenyl; R.sub.1 and R.sub.2 are both
o-CH.sub.3C(O)O-phenyl, and R.sub.3 and R.sub.4 are phenyl; R.sub.1
and R.sub.2 are both phenyl, and R.sub.3 and R.sub.4 are both
methyl; R.sub.1 and R.sub.2 are both phenyl, and R.sub.3 and
R.sub.4 are both ethyl; R.sub.1 and R.sub.2 are both phenyl, and
R.sub.3 and R.sub.4 are both n-propyl; R.sub.1 and R.sub.2 are both
p-cyanophenyl, and R.sub.3 and R.sub.4 are both methyl; R.sub.1 and
R.sub.2 are both p-nitro phenyl, and R.sub.3 and R.sub.4 are both
methyl; R.sub.1 and R.sub.2 are both 2,5-dimethoxyphenyl, and
R.sub.3 and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both
phenyl, and R.sub.3 and R.sub.4 are both n-butyl; R.sub.1 and
R.sub.2 are both p-chlorophenyl, and R.sub.3 and R.sub.4 are both
methyl; R.sub.1 and R.sub.2 are both 3-nitrophenyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both
3-cyanophenyl, and R.sub.3 and R.sub.4 are both methyl; R.sub.1 and
R.sub.2 are both 3-fluorophenyl, and R.sub.3 and R.sub.4 are both
methyl; R.sub.1 and R.sub.2 are both 2-furanyl, and R.sub.3 and
R.sub.4 are both phenyl; R.sub.1 and R.sub.2 are both
2-methoxyphenyl, and R.sub.3 and R.sub.4 are both methyl; R.sub.1
and R.sub.2 are both 3-methoxyphenyl, and R.sub.3 and R.sub.4 are
both methyl; R.sub.1 and R.sub.2 are both 2,3-dimethoxyphenyl, and
R.sub.3 and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both
2-methoxy-5-chlorophenyl, and R.sub.3 and R.sub.4 are both ethyl;
R.sub.1 and R.sub.2 are both 2,5-difluorophenyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both
2,5-dichlorophenyl, and R.sub.3 and R.sub.4 are both methyl;
R.sub.1 and R.sub.2 are both 2,5-dimethylphenyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both
2-methoxy-5-chlorophenyl, and R.sub.3 and R.sub.4 are both methyl;
R.sub.1 and R.sub.2 are both 3,6-dimethoxyphenyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both phenyl, and
R.sub.3 and R.sub.4 are both 2-ethylphenyl; R.sub.1 and R.sub.2 are
both 2-methyl-5-pyridyl, and R.sub.3 and R.sub.4 are both methyl;
or R.sub.1 is phenyl; R.sub.2 is 2,5-dimethoxyphenyl, and R.sub.3
and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both methyl,
and R.sub.3 and R.sub.4 are both p-CF.sub.3-phenyl; R.sub.1 and
R.sub.2 are both methyl, and R.sub.3 and R.sub.4 are both
o-CH.sub.3-phenyl; R.sub.1 and R.sub.2 are both
--(CH.sub.2).sub.3COOH; and R.sub.3 and R.sub.4 are both phenyl;
R.sub.1 and R.sub.2 are both represented by the following
structural formula
##STR00007##
and R.sub.3 and R.sub.4 are both phenyl; R.sub.1 and R.sub.2 are
both n-butyl, and R.sub.3 and R.sub.4 are both phenyl; R.sub.1 and
R.sub.2 are both n-pentyl, R.sub.3 and R.sub.4 are both phenyl;
R.sub.1 and R.sub.2 are both methyl, and R.sub.3 and R.sub.4 are
both 2-pyridyl; R.sub.1 and R.sub.2 are both cyclohexyl, and
R.sub.3 and R.sub.4 are both phenyl; R.sub.1 and R.sub.2 are both
methyl, and R.sub.3 and R.sub.4 are both 2-ethylphenyl; R.sub.1 and
R.sub.2 are both methyl, and R.sub.3 and R.sub.4 are both
2,6-dichlorophenyl; R.sub.1-R.sub.4 are all methyl; R.sub.1 and
R.sub.2 are both methyl, and R.sub.3 and R.sub.4 are both t-butyl;
R.sub.1 and R.sub.2 are both ethyl, and R.sub.3 and R.sub.4 are
both methyl; R.sub.1 and R.sub.2 are both t-butyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both cyclopropyl,
and R.sub.3 and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are
both cyclopropyl, and R.sub.3 and R.sub.4 are both ethyl; R.sub.1
and R.sub.2 are both 1-methylcyclopropyl, and R.sub.3 and R.sub.4
are both methyl; R.sub.1 and R.sub.2 are both 2-methylcyclopropyl,
and R.sub.3 and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are
both 1-phenylcyclopropyl, and R.sub.3 and R.sub.4 are both methyl;
R.sub.1 and R.sub.2 are both 2-phenylcyclopropyl, and R.sub.3 and
R.sub.4 are both methyl; R.sub.1 and R.sub.2 are both cyclobutyl,
and R.sub.3 and R.sub.4 are both methyl; R.sub.1 and R.sub.2 are
both cyclopentyl, and R.sub.3 and R.sub.4 are both methyl; R.sub.1
is cyclopropyl, R.sub.2 is phenyl, and R.sub.3 and R.sub.4 are both
methyl.
[0050] Preferred examples of bis(thiohydrazide amides) include
Compounds (1)-(18) and pharmaceutically acceptable salts and
solvates thereof:
##STR00008## ##STR00009## ##STR00010##
[0051] Particular examples of bis(thiohydrazide amides) include
Compounds (1), (17), and (18) and pharmaceutically acceptable salts
and solvates thereof.
[0052] The taxanes employed in the disclosed invention include
Taxol.TM. and Taxol.TM. analogs. Taxol.TM. or "paclitaxel" is a
well-known anti-cancer drug which can act by enhancing and
stabilizing microtubule formation. Thus, the term "Taxol.TM.
analog" is defined herein to mean a compound which has the basic
Taxol.TM. skeleton and which stabilizes microtubule formation. Many
analogs of Taxol.TM. are known, including Taxotere.TM., also
referred to as "docetaxol". Taxol.TM. and Taxotere.TM. have the
respective structural formulas:
##STR00011##
[0053] The taxanes employed in the disclosed invention have the
basic taxane skeleton as a common structure feature shown below in
Structural Formula VI:
##STR00012##
Double bonds have been omitted from the cyclohexane rings in the
taxane skeleton represented by Structural Formula VI. It is to be
understood that the basic taxane skeleton can include zero or one
double bond in one or both cyclohexane rings, as indicated in the
Taxol.TM. analogs and Structural Formulas VII and VIII below. A
number of atoms have also been omitted from Structural Formula VI
to indicate sites in which structural variation commonly occurs
among Taxol.TM. analogs.
[0054] A wide variety of substituents can decorate the taxane
skeleton without adversely affecting biological activity. Also,
zero, one or both of the cyclohexane rings of a Taxol.TM. analog
can have a double bond at the indicated positions. For example,
substitution on the taxane skeleton with simply an oxygen atom
indicates that hydroxyl, acyl, alkoxy or other oxygen-bearing
substituent is commonly found at the site. It is to be understood
that these and other substitutions on the taxane skeleton can be
made without losing the ability to enhance and stabilize
microtubule formation. Thus, the term "Taxol.TM. analog" is defined
herein to mean a compound which has the basic Taxol.TM. skeleton
and which stabilizes microtubule formation. The term taxane is
defined herein to include compounds such as Taxol.TM. and the
"Taxol.TM. analogs" described herein, or a pharmaceutically
acceptable salt or solvate thereof.
[0055] Typically, the taxanes employed in the disclosed invention
are represented by Structural Formula VII or VIII:
##STR00013##
[0056] R.sub.10 is an optionally substituted lower alkyl group, an
optionally substituted phenyl group, --SR.sub.19, --NHR.sub.19 or
--OR.sub.19.
[0057] R.sub.11 is an optionally substituted lower alkyl group, an
optionally substituted aryl group.
[0058] R.sub.12 is --H, --OH, lower alkyl, substituted lower alkyl,
lower alkoxy, substituted lower alkoxy, --O--C(O)-(lower alkyl),
--O--C(O)-(substituted lower alkyl), --O--CH.sub.2--O-(lower
alkyl)-S--CH.sub.2--O-(lower alkyl).
[0059] R.sub.13 is --H, --CH.sub.3, or, taken together with
R.sub.14, --CH.sub.2--.
[0060] R.sub.14 is --H, --OH, lower alkoxy, --O--C(O)-(lower
alkyl), substituted lower alkoxy, --O--C(O)-(substituted lower
alkyl), --O--CH.sub.2--O--P(O)(OH).sub.2, --O--CH.sub.2--O-(lower
alkyl), --O--CH.sub.2--S-(lower alkyl) or, taken together with
R.sub.20, a double bond.
[0061] R.sub.15--H, lower acyl, lower alkyl, substituted lower
alkyl, alkoxymethyl, alkthiomethyl, --OC(O)--O(lower alkyl),
--OC(O)--O(substituted lower alkyl), --OC(O)--NH(lower alkyl) or
--OC(O)--NH(substituted lower alkyl).
[0062] R.sub.16 is phenyl or substituted phenyl.
[0063] R.sub.17 is --H, lower acyl, substituted lower acyl, lower
alkyl, substituted, lower alkyl, (lower alkoxy)methyl or (lower
alkyl)thiomethyl.
[0064] R.sub.18--H, --CH.sub.3 or, taken together with R.sub.17 and
the carbon atoms to which R.sub.17 and
[0065] R.sub.18 are bonded, a five or six membered a non-aromatic
heterocyclic ring.
[0066] R.sub.19 is an optionally substituted lower alkyl group, an
optionally substituted phenyl group.
[0067] R.sub.20 is --H or a halogen.
[0068] R.sub.21 is --H, lower alkyl, substituted lower alkyl, lower
acyl or substituted lower acyl.
[0069] Preferably, the variables in Structural Formulas VII and
VIII are defined as follows: R.sub.10 is phenyl, tert-butoxy,
--S--CH.sub.2--CH--(CH.sub.3).sub.2, --S--CH(CH.sub.3).sub.3,
--S--(CH.sub.2).sub.3CH.sub.3, --O--CH(CH.sub.3).sub.3,
--NH--CH(CH.sub.3).sub.3, --CH.dbd.C(CH.sub.3).sub.2 or
para-chlorophenyl; R.sub.u is phenyl, (CH.sub.3).sub.2CHCH.sub.2--,
-2-furanyl, cyclopropyl or para-toluoyl; R.sub.12 is --H, --OH,
CH.sub.3CO-- or --(CH.sub.2).sub.2--N-morpholino; R.sub.13 is
methyl, or, R.sub.13 and R.sub.14, taken together, are
--CH.sub.2--;
[0070] R.sub.14 is --H, --CH.sub.2SCH.sub.3 or
--CH.sub.2--O--P(O)(OH).sub.2; R.sub.15 is CH.sub.3CO--;
[0071] R.sub.16 is phenyl; R.sub.17--H, or, R.sub.17 and R.sub.18,
taken together, are --O--CO--O--;
[0072] R.sub.18 is --H; R.sub.20 is --H or --F; and R.sub.21 is
--H, --C(O)--CHBr--(CH.sub.2).sub.13--CH.sub.3 or
--C(O)--(CH.sub.2).sub.14--CH.sub.3;
--C(O)--CH.sub.2--CH(OH)--COOH,
--C(O)--CH.sub.2--O--C(O)--CH.sub.2CH(NH.sub.2)--CONH.sub.2,
--C(O)--CH.sub.2--O--CH.sub.2CH.sub.2OCH.sub.3 or
--C(O)--O--C(O)--CH.sub.2CH.sub.3.
[0073] Specific examples of Taxol.TM. analogs include the following
compounds:
##STR00014## ##STR00015## ##STR00016## ##STR00017## ##STR00018##
##STR00019##
[0074] A Taxol.TM. analog can also be bonded to or be pendent from
a pharmaceutically acceptable polymer, such as a polyacrylamide.
One example of a polymer of this type is Taxol.TM. analog 22,
below, which has the structure of a polymer comprising a taxol
analog group pendent from the polymer backbone. The polymer is a
terpolymer of the three monomer units shown. The term "Taxol.TM.
analog", as it is used herein, includes such polymers.
##STR00020##
[0075] A "straight chained hydrocarbyl group" is an alkylene group,
i.e., --(CH.sub.2).sub.y--, with one, or more (preferably one)
internal methylene groups optionally replaced with a linkage group.
y is a positive integer (e.g., between 1 and 10), preferably
between 1 and 6 and more preferably 1 or 2. A "linkage group"
refers to a functional group which replaces a methylene in a
straight chained hydrocarbyl. Examples of suitable linkage groups
include a ketone (--C(O)--), alkene, alkyne, phenylene, ether
(--O--), thioether (--S--), or amine (--N(R.sup.a)--), wherein
R.sup.a is defined below. A preferred linkage group is
--C(R.sub.5R.sub.6)--, wherein R.sub.5 and R.sub.6 are defined
above. Suitable substituents for an alkylene group and a
hydrocarbyl group are those which do not substantially interfere
with the anti-cancer activity of the bis(thiohydrazide) amides and
taxanes. R.sub.5 and R.sub.6 are preferred substituents for an
alkylene or hydrocarbyl group represented by Y.
[0076] An aliphatic group is a straight chained, branched or cyclic
non-aromatic hydrocarbon which is completely saturated or which
contains one or more units of unsaturation. Typically, a straight
chained or branched aliphatic group has from 1 to about 20 carbon
atoms, preferably from 1 to about 10, and a cyclic aliphatic group
has from 3 to about 10 carbon atoms, preferably from 3 to about 8.
An aliphatic group is preferably a straight chained or branched
alkyl group, e.g., methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, tert-butyl, pentyl, hexyl, pentyl or octyl, or a
cycloalkyl group with 3 to about 8 carbon atoms. A C1-C20 straight
chained or branched alkyl group or a C3-C8 cyclic alkyl group is
also referred to as a "lower alkyl" group.
[0077] The term "aromatic group" may be used interchangeably with
"aryl," "aryl ring," "aromatic ring," "aryl group" and "aromatic
group." Aromatic groups include carbocyclic aromatic groups such as
phenyl, naphthyl, and anthracyl, and heteroaryl groups such as
imidazolyl, thienyl, furanyl, pyridyl, pyrimidyl, pyranyl,
pyrazolyl, pyrroyl, pyrazinyl, thiazole, oxazolyl, and tetrazole.
The term "heteroaryl group" may be used interchangeably with
"heteroaryl," "heteroaryl ring," "heteroaromatic ring" and
"heteroaromatic group." The term "heteroaryl," as used herein,
means a mono- or multi-cyclic aromatic heterocycle which comprise
at least one heteroatom such as nitrogen, sulfur and oxygen, but
may include 1, 2, 3 or 4 heteroatoms per ring. Aromatic groups also
include fused polycyclic aromatic ring systems in which a
carbocyclic aromatic ring or heteroaryl ring is fused to one or
more other heteroaryl rings. Examples include benzothienyl,
benzofuranyl, indolyl, quinolinyl, benzothiazole, benzooxazole,
benzimidazole, quinolinyl, isoquinolinyl and isoindolyl.
[0078] The term "arylene" refers to an aryl group which is
connected to the remainder of the molecule by two other bonds. By
way of example, the structure of a 1,4-phenylene group is shown
below:
##STR00021##
[0079] Substituents for an arylene group are as described below for
an aryl group.
[0080] Non-aromatic heterocyclic rings are non-aromatic rings which
include one or more heteroatoms such as nitrogen, oxygen or sulfur
in the ring. The ring can be five, six, seven or eight-membered.
Examples include tetrahydrofuranyl, tetrahydrothiophenyl,
morpholino, thiomorpholino, pyrrolidinyl, piperazinyl, piperidinyl,
and thiazolidinyl.
[0081] Suitable substituents on an aliphatic group (including an
alkylene group), non-aromatic heterocyclic group, benzylic or aryl
group (carbocyclic and heteroaryl) are those which do not
substantially interfere with the anti-cancer activity of the
bis(thiohydrazide) amides and taxanes. A substituent substantially
interferes with anti-cancer activity when the anti-cancer activity
is reduced by more than about 50% in a compound with the
substituent compared with a compound without the substituent.
Examples of suitable substituents include --R.sup.a, --OH, --Br,
--Cl, --I, --F, --OR.sup.a, --O--COR.sup.a, --COR.sup.a, --CN,
--NO.sub.2, --COOH, --SO.sub.3H, --NH.sub.2, --NHR.sup.a,
--N(R.sup.aR.sup.b), --COOR.sup.a, --CHO, --CONH.sub.2,
--CONHR.sup.a, --CON(R.sup.aR.sup.b), --NHCOR.sup.a,
--NR.sup.cCOR.sup.a, --NHCONH.sub.2, --NHCONR.sup.aH,
--NHCON(R.sup.aR.sup.b), --NR.sup.cCONH.sub.2,
--NR.sup.cCONR.sup.aH, --NR.sup.cCON(R.sup.aR.sup.b),
--C(.dbd.NH)--NH.sub.2, --C(.dbd.NH)--NHR.sup.a,
--C(.dbd.NH)--N(R.sup.aR.sup.b), --C(.dbd.NR.sup.c)--NH.sub.2,
--C(.dbd.NR.sup.c)--NHR.sup.a,
--C(.dbd.NR.sup.c)--N(R.sup.aR.sup.b), --NH--C(.dbd.NH)--NH.sub.2,
--NH--C(.dbd.NH)--NHR.sup.a, --NH--C(.dbd.NH)--N(R.sup.aR.sup.b),
--NH--C(.dbd.NR.sup.c)--NH.sub.2,
--NH--C(.dbd.NR.sup.c)--NHR.sup.a,
--NH--C(.dbd.NR.sup.c)--N(R.sup.aR.sup.b),
--NR.sup.dH--C(.dbd.NH)--NH.sub.2,
--NR.sup.d--C(.dbd.NH)--NHR.sup.a,
--NR.sup.d--C(.dbd.NH)--N(R.sup.aR.sup.b),
--NR.sup.d--C(.dbd.NR.sup.c)--NH.sub.2,
--NR.sup.d--C(.dbd.NR.sup.c)--NHR.sup.a,
--NR.sup.d--C(.dbd.NR.sup.c)--N(R.sup.aR.sup.b), --NHNH.sub.2,
--NHNHR.sup.a, --NHR.sup.aR.sup.b, --SO.sub.2NH.sub.2,
--SO.sub.2NHR.sup.a, --SO.sub.2NR.sup.aR.sup.b, --CH.dbd.CHR.sup.a,
--CH.dbd.CR.sup.aR.sup.b, --CR.sup.c.dbd.CR.sup.aR.sup.b,
--CR.sup.c.dbd.CHR.sup.a, --CR.sup.c.dbd.CR.sup.aR.sup.b,
--CCR.sup.a, --SH, --SR.sup.a, --S(O)R.sup.a, --S(O).sub.2R.sup.a.
R.sup.a--R.sup.d are each independently an alkyl group, aromatic
group, non-aromatic heterocyclic group or --N(R.sup.aR.sup.b),
taken together, form an optionally substituted non-aromatic
heterocyclic group. The alkyl, aromatic and non-aromatic
heterocyclic group represented by R.sup.a--R.sup.d and the
non-aromatic heterocyclic group represented by --N(R.sup.aR.sup.b)
are each optionally and independently substituted with one or more
groups represented by R.sup.#.
[0082] R.sup.# is R.sup.+, --OR.sup.+, --O(haloalkyl), --SR.sup.+,
--NO.sub.2, --CN, --NCS, --N(R.sup.+).sub.2, --NHCO.sub.2R.sup.+,
--NHC(O)R.sup.+, --NHNHC(O)R.sup.+, --NHC(O)N(R.sup.+).sub.2,
--NHNHC(O)N(R.sup.+).sub.2, --NHNHCO.sub.2R.sup.+,
--C(O)C(O)R.sup.+, --C(O)CH.sub.2C(O)R.sup.+, --CO.sub.2R.sup.+,
--C(O)R.sup.+, --C(O)N(R.sup.+).sub.2, --OC(O)R.sup.+,
--OC(O)N(R.sup.+).sub.2, --S(O).sub.2R.sup.+,
--SO.sub.2N(R.sup.+).sub.2, --S(O)R.sup.+,
--NHSO.sub.2N(R.sup.+).sub.2, --NHSO.sub.2R.sup.+,
--C(.dbd.S)N(R.sup.+).sub.2, or --C(.dbd.NH)--N(R.sup.+).sub.2.
[0083] R.sup.+ is --H, a C1-C4 alkyl group, a monocyclic heteroaryl
group, a non-aromatic heterocyclic group or a phenyl group
optionally substituted with alkyl, haloalkyl, alkoxy, haloalkoxy,
halo, --CN, --NO.sub.2, amine, alkylamine or dialkylamine.
Optionally, the group --N(R.sup.+).sub.2 is a non-aromatic
heterocyclic group, provided that non-aromatic heterocyclic groups
represented by R.sup.+ and --N(R.sup.+).sub.2 that comprise a
secondary ring amine are optionally acylated or alkylated.
[0084] Preferred substituents for a phenyl group, including phenyl
groups represented by R.sub.1-R.sub.4, include C1-C4 alkyl, C1-C4
alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, phenyl, benzyl, pyridyl,
--OH, --NH.sub.2, --F, --Cl, --Br, --I, --NO.sub.2 or --CN.
[0085] Preferred substituents for an aliphatic group, including
aliphatic groups represented by R.sub.1-R.sub.4, include C1-C4
alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, phenyl,
benzyl, pyridyl, --OH, --NH.sub.2, --F, --Cl, --Br, --I, --NO.sub.2
or --CN.
[0086] Preferred substituents for a cycloalkyl group, including
cycloalkyl groups represented by R.sub.1 and R.sub.2, are alkyl
groups, such as a methyl or ethyl groups.
[0087] Also included in the present invention are pharmaceutically
acceptable salts of the bis(thiohydrazide) amides and taxanes
employed herein. These compounds can have one or more sufficiently
acidic protons that can react with a suitable organic or inorganic
base to form a base addition salt. Base addition salts include
those derived from inorganic bases, such as ammonium or alkali or
alkaline earth metal hydroxides, carbonates, bicarbonates, and the
like, and organic bases such as alkoxides, alkyl amides, alkyl and
aryl amines, and the like. Such bases useful in preparing the salts
of this invention thus include sodium hydroxide, potassium
hydroxide, ammonium hydroxide, potassium carbonate, and the
like.
[0088] For example, pharmaceutically acceptable salts of
bis(thiohydrazide) amides and taxanes employed herein (e.g., those
represented by Structural Formulas I-VI, Compounds 1-18, and
Taxol.TM. analogs 1-22) are those formed by the reaction of the
compound with one equivalent of a suitable base to form a
monovalent salt (i.e., the compound has single negative charge that
is balanced by a pharmaceutically acceptable counter cation, e.g.,
a monovalent cation) or with two equivalents of a suitable base to
form a divalent salt (e.g., the compound has a two-electron
negative charge that is balanced by two pharmaceutically acceptable
counter cations, e.g., two pharmaceutically acceptable monovalent
cations or a single pharmaceutically acceptable divalent cation).
Divalent salts of the bis(thiohydrazide amides) are preferred.
"Pharmaceutically acceptable" means that the cation is suitable for
administration to a subject. Examples include Li.sup.+, Na.sup.+,
K.sup.+, Mg.sup.2+, Ca.sup.2+ and NR.sub.4.sup.+, wherein each R is
independently hydrogen, an optionally substituted aliphatic group
(e.g., a hydroxyalkyl group, aminoalkyl group or ammoniumalkyl
group) or optionally substituted aryl group, or two R groups, taken
together, form an optionally substituted non-aromatic heterocyclic
ring optionally fused to an aromatic ring. Generally, the
pharmaceutically acceptable cation is Li.sup.+, Na.sup.+, K.sup.+,
NH.sub.3(C.sub.2H.sub.5OH).sup.+ or
N(CH.sub.3).sub.3(C.sub.2H.sub.5OH).sup.+, and more typically, the
salt is a disodium or dipotassium salt, preferably the disodium
salt.
[0089] Bis(thiohydrazide) amides and taxanes employed herein having
a sufficiently basic group, such as an amine can react with an
organic or inorganic acid to form an acid addition salt. Acids
commonly employed to form acid addition salts from compounds with
basic groups are inorganic acids such as hydrochloric acid,
hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid,
and the like, and organic acids such as p-toluenesulfonic acid,
methanesulfonic acid, oxalic acid, p-bromophenyl-sulfonic acid,
carbonic acid, succinic acid, citric acid, benzoic acid, acetic
acid, and the like. Examples of such salts include the sulfate,
pyrosulfate, bisulfate, sulfite, bisulfite, phosphate,
monohydrogenphosphate, dihydrogenphosphate, metaphosphate,
pyrophosphate, chloride, bromide, iodide, acetate, propionate,
decanoate, caprylate, acrylate, formate, isobutyrate, caproate,
heptanoate, propiolate, oxalate, malonate, succinate, suberate,
sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate,
benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate,
hydroxybenzoate, methoxybenzoate, phthalate, sulfonate,
xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate,
citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate,
methanesulfonate, propanesulfonate, naphthalene-1-sulfonate,
naphthalene-2-sulfonate, mandelate, and the like.
[0090] Particular salts of the bis(thiohydrazide amide) compounds
described herein can be prepared according to methods described in
copending, co-owned Patent Application Ser. No. 60/582,596, filed
Jun. 23, 2004.
[0091] The neutral bis(thiohydrazide) amides can be prepared
according to methods described in U.S. Pat. Nos. 6,800,660, and
6,762,204, both entitled "Synthesis of Taxol Enhancers" and also
according to methods described in the co-pending and co-owned U.S.
patent application Ser. Nos. 10/345,885 filed Jan. 15, 2003, and
10/758,589, Jan. 15, 2004. The entire teachings of each document
referred to in this application is expressly incorporated herein by
reference.
[0092] It will also be understood that certain compounds employed
in the invention may be obtained as different stereoisomers (e.g.,
diastereomers and enantiomers) and that the invention includes all
isomeric forms and racemic mixtures of the disclosed compounds and
methods of treating a subject with both pure isomers and mixtures
thereof, including racemic mixtures. Stereoisomers can be separated
and isolated using any suitable method, such as chromatography.
[0093] As used herein, a "subject" is a mammal, preferably a human,
but can also be an animal in need of veterinary treatment, e.g.,
companion animals (e.g., dogs, cats, and the like), farm animals
(e.g., cows, sheep, pigs, horses, and the like) and laboratory
animals (e.g., rats, mice, guinea pigs, and the like).
[0094] The bis(thiohydrazide) amides and taxanes employed herein
can be administered to a subject by any conventional method of drug
administration for treatment of cancerous disorders, for example,
orally in capsules, suspensions or tablets or by parenteral
administration. Parenteral administration can include, for example,
systemic administration, such as by intramuscular, intravenous,
subcutaneous, or intraperitoneal injection. In specific
embodiments, oral, parenteral, or local administration are
preferred modes of administration for treatment of cancer.
Preferably, the mode of administration is intravenous.
[0095] An effective amount of a bis(thio-hydrazide) amide or a
taxane anticancer compound is a quantity in which anti-cancer
effects are normally achieved. With respect to a particular
compound in the method (e.g., the bis(thio-hydrazide) amide or the
taxane anticancer compound), an "effective amount" is the quantity
in which a greater anti-cancer effect is achieved when the
particular compound is co-administered with the other compounds in
the method compared with when the particular compound is
administered alone. The compounds of the method can be
co-administered to the subject as part of the same pharmaceutical
composition or, alternatively, as separate pharmaceutical
compositions. When administered as separate pharmaceutical
compositions, the compounds of the method can be administered
simultaneously or at different times, provided that the enhancing
effect of the compounds in combination is retained.
[0096] As used herein, "treating a subject with a cancer," or
similar terms, includes achieving, partially or substantially, one
or more of the following: arresting the growth or spread of a
cancer, reducing the extent of a cancer (e.g., reducing size of a
tumor or reducing the number of affected sites), inhibiting the
growth rate of a cancer, and ameliorating or improving a clinical
symptom or indicator associated with a cancer (such as tissue or
serum components).
[0097] In various embodiments, cancer can include human sarcomas
and carcinomas, e.g., fibrosarcoma, myxo sarcoma, liposarcoma,
chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,
endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,
synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,
rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast
cancer, ovarian cancer, prostate cancer, squamous cell carcinoma,
basal cell carcinoma, adenocarcinoma, sweat gland carcinoma,
sebaceous gland carcinoma, papillary carcinoma, papillary
adenocarcinomas, cystadenocarcinoma, medullary carcinoma,
bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct
carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms'
tumor, cervical cancer, testicular tumor, lung carcinoma, small
cell lung carcinoma, bladder carcinoma, epithelial carcinoma,
glioma, astrocytoma, medulloblastoma, craniopharyngioma,
ependymoma, pinealoma, hemangioblastoma, acoustic neuroma,
oligodendroglioma, meningioma, melanoma, neuroblastoma,
retinoblastoma; leukemias, e.g., acute lymphocytic leukemia and
acute myelocytic leukemia (myeloblastic, promyelocytic,
myelomonocytic, monocytic and erythroleukemia); chronic leukemia
(chronic myelocytic (granulocytic) leukemia and chronic lymphocytic
leukemia); and polycythemia vera, lymphoma (Hodgkin's disease and
non-Hodgkin's disease), multiple myeloma, Waldenstrobm's
macroglobulinemia, and heavy chain disease.
[0098] In some embodiments, cancer can include leukemias e.g.,
acute and/or chronic leukemias, e.g., lymphocytic leukemia (e.g.,
as exemplified by the p388 (murine) cell line), large granular
lymphocytic leukemia, and lymphoblastic leukemia; T-cell leukemias,
e.g., T-cell leukemia (e.g., as exemplified by the CEM, Jurkat, and
HSB-2 (acute), YAC-1 (murine) cell lines), T-lymphocytic leukemia,
and T-lymphoblastic leukemia; B cell leukemia (e.g., as exemplified
by the SB (acute) cell line), and B-lymphocytic leukemia; mixed
cell leukemias, e.g., B and T cell leukemia and B and T lymphocytic
leukemia; myeloid leukemias, e.g., granulocytic leukemia,
myelocytic leukemia (e.g., as exemplified by the HL-60
(promyelocyte) cell line), and myelogenous leukemia (e.g., as
exemplified by the K562(chronic)cell line); neutrophilic leukemia;
eosinophilic leukemia; monocytic leukemia (e.g., as exemplified by
the THP-1 (acute) cell line); myelomonocytic leukemia; Naegeli-type
myeloid leukemia; and nonlymphocytic leukemia. Other examples of
leukemias are described in Chapter 60 of The Chemotherapy
Sourcebook, Michael C. Perry Ed., Williams & Williams (1992)
and Section 36 of Holland Frie Cancer Medicine 5th Ed., Bast et al.
Eds., B.C. Decker Inc. (2000). The entire teachings of the
preceding references are incorporated herein by reference.
[0099] In certain embodiments, cancer can include non-solid tumors
such as multiple myeloma, T-leukemia (e.g., as exemplified by
Jurkat and CEM cell lines); B-leukemia (e.g., as exemplified by the
SB cell line); promyelocytes (e.g., as exemplified by the HL-60
cell line); uterine sarcoma (e.g., as exemplified by the MES-SA
cell line); monocytic leukemia (e.g., as exemplified by the THP-1
(acute) cell line); and lymphoma (e.g., as exemplified by the U937
cell line).
[0100] In some embodiments, cancer can include colon cancer,
pancreatic cancer, melanoma, renal cancer, sarcoma, breast cancer,
ovarian cancer, lung cancer, stomach cancer, bladder cancer and
cervical cancer.
[0101] In some embodiments, the disclosed methods can be
particularly effective at treating subjects whose cancer has become
"multi-drug resistant". A cancer which initially responded to an
anti-cancer drug becomes resistant to the anti-cancer drug when the
anti-cancer drug is no longer effective in treating the subject
with the cancer. For example, many tumors can initially respond to
treatment with an anti-cancer drug by decreasing in size or even
going into remission, only to develop resistance to the drug. Drug
resistant tumors are characterized by a resumption of their growth
and/or reappearance after having seemingly gone into remission,
despite the administration of increased dosages of the anti-cancer
drug. Cancers that have developed resistance to two or more
anti-cancer drugs are said to be "multi-drug resistant". For
example, it is common for cancers to become resistant to three or
more anti-cancer agents, often five or more anti-cancer agents and
at times ten or more anti-cancer agents.
[0102] The bis(thiohydrazide) amides and taxanes employed herein
can be administered to the subject in conjunction with an
acceptable pharmaceutical carrier or diluent as part of a
pharmaceutical composition for treatment cancer therapy.
Formulation of the compound to be administered will vary according
to the route of administration selected (e.g., solution, emulsion,
capsule, and the like). Suitable pharmaceutically acceptable
carriers may contain inert ingredients which do not unduly inhibit
the biological activity of the compounds. The pharmaceutically
acceptable carriers should be biocompatible, i.e., non-toxic,
non-inflammatory, non-immunogenic and devoid of other undesired
reactions upon the administration to a subject. Standard
pharmaceutical formulation techniques can be employed, such as
those described in Remington's Pharmaceutical Sciences, ibid.
Suitable pharmaceutical carriers for parenteral administration
include, for example, sterile water, physiological saline,
bacteriostatic saline (saline containing about 0.9% mg/ml benzyl
alcohol), phosphate-buffered saline, Hank's solution,
Ringer's-lactate and the like. Methods for encapsulating
compositions (such as in a coating of hard gelatin or cyclodextran)
are known in the art (Baker, et al., "Controlled Release of
Biological Active Agents", John Wiley and Sons, 1986).
[0103] In certain embodiments, one or more compounds of the
invention and one or more other the therapies (e.g., prophylactic
or therapeutic agents) are cyclically administered. Cycling therapy
involves the administration of a first therapy (e.g., a first
prophylactic or therapeutic agents) for a period of time, followed
by the administration of a second therapy (e.g., a second
prophylactic or therapeutic agents) for a period of time, followed
by the administration of a third therapy (e.g., a third
prophylactic or therapeutic agents) for a period of time and so
forth, and repeating this sequential administration, i.e., the
cycle in order to reduce the development of resistance to one of
the agents, to avoid or reduce the side effects of one of the
agents, and/or to improve the efficacy of the treatment.
[0104] In various embodiments, the methods herein can include
administration prior to or concurrently with the bis(thiohydrazide)
amide/taxane combination, agents that can reduce acute irritation
or allergic reaction to administration, e.g., an anti-inflammatory
such as Decadron.RTM. (dexamethasone, e.g., 10 mg intravenously),
an antihistamine such as Benadryl.RTM. (diphenhydramine, e.g., 50
mg intravenously), an antacid such as Zantac.RTM. (ranitidine
hydrochloride, e.g., 50 mg intravenously), and the like.
EXEMPLIFICATION
Example 1
Measurement of Heat Shock Protein 70 (Hsp70)
[0105] Plasma Hsp70 was measured by a sandwich ELISA kit (Stressgen
Bioreagents Victoria, British Columbia, CANADA) according to a
modified protocol in house. In brief, Hsp70 in plasma specimens and
serial concentrations of Hsp70 standard were captured onto 96-well
plate on which anti-Hsp70 antibody was coated. Then captured Hsp70
was detected with a biotinylated anti-Hsp70 antibody followed by
incubation with europium-conjugated streptavidin. After each
incubation unbound materials were removed by washing. Finally,
antibody-Hsp70 complex was measured by time resolved fluorometry of
europium. Concentration of Hsp70 was calculated from a standard
curve.
Example 2
Measurement of Natural Killer Cell Cytotoxic Activity
[0106] The following procedure can be employed to assay NK cell
activity in a subject. The procedure is adapted from Kantakamalakul
W, Jaroenpool J, Pattanapanyasat K. A novel enhanced green
fluorescent protein (EGFP)-K562 flow cytometric method for
measuring natural killer (NK) cell cytotoxic activity. J Immunol
Methods. 2003 Jan. 15; 272:189-197, the entire teachings of which
are incorporated herein by reference.
[0107] Materials and methods: Human erythroleukaemic cell line,
K562, was obtained from American Type Culture Collection (CCL-243,
American Type Culture Collection, Manassas, Va.), and cultured in
RPMI-1640 medium (Cat#11875-093 Gibco Invitrogen Corp, Carlsbad,
Calif.) supplemented with 10% heat inactivated fetal calf serum
(Gibco), 2 mM L-glutamin, 100 .mu.g/ml streptomycin and 100 IU/ml
penicillin at 37.degree. C. with 5% CO.sub.2. K562 cells were
transduced with retroviral vector which encode green fluorescent
protein (eGFP). Stable cell line was selected with antibiotic,
G418. About 99.6% G418 resistant cells were eGFP positive after
section.
[0108] The subject's peripheral blood mononuclear cells (PBMCs)
were prepared by clinical study sites and received in BD Vacutainer
Cell Preparation Tube with sodium heparin (Product Number: 362753,
Becton Dickinson, Franklin Lakes, N.J.).
[0109] Two-fold serial dilution of 800 .mu.l effector cells
(patient's PBMC) starting at concentration of 1.times.10.sup.6
cells/mL were put into four individual polystyrene 12.times.75-mm
tubes. Log phase growing target cells (K562/eGFP) were adjusted
with growth medium (RPMI-1640) to a concentration of
1.times.10.sup.5 cells/mL and 100 .mu.L targets then added into the
tubes to provide effector/target (E/T) ratios of 80:1, 40:1, 20:1,
10:1. Effector cells alone and target cells alone were used as
controls. All tubes were incubated at 37.degree. C. with 5%
CO.sub.2 for about 3.5 hr. Ten microliters of propidium iodide (PI)
at a concentration of 1 mg/mL was added to each tube including
effector and target control tubes and then incubated at room
temperature for 15 min.
[0110] Cytotoxic activity was analyzed with a FACSCalibur flow
cytometer (Becton Dickinson). Linear amplification of the forward
and side scatter (FSC/SSC) signals, as well as logarithmic
amplification of eGFP and PI emission in green and red fluorescence
were obtained. Ten thousand events per sample tube with no gating
for acquisition were collected for analysis. Data analysis for
two-parameter dot plots for eGFP versus PI was performed using
CELLQuest (Becton Dickinson Biosciences) software to enumerate live
and dead target cells. Debris and dead cells were excluded by
setting a threshold of forward light scatter.
Example 3
The Disclosed Combination Therapy Induces Hsp70
[0111] A Phase I trial was conducted for combined administration of
a bis(thio-hydrazide) amide (Compound (1)) and a taxane
(paclitaxel) to human subjects with various advanced solid tumors.
Compound (1) and paclitaxel were co-administered intravenously over
3 hours every 3 weeks. Starting doses were 44
milligrams/meter.sup.2 (mg/m2, or 110 micromoles/meter.sup.2
(.mu.mol/m2)) Compound (1) and 135 mg/m2 (158 .mu.mol/m2)
paclitaxel. Paclitaxel was then increased to 175 mg/m2 (205
.mu.mol/m2), followed by escalation of Compound (1) to establish
the maximum tolerated dose based on first cycle toxicity in 3 to 6
patients at each dose level. Pharmacokinetic (PK) studies were
performed during cycle 1 using liquid chromatography/mass
spectrometry (LC/MS) to measure both compounds in plasma. Heat
shock protein 70 (Hsp70) was measured in plasma before and after
treatment. 35 patients were evaluated at 8 dose levels, including
paclitaxel at 135 mg/m2 (158 .mu.mol/m2) and Compound (1) at 44
mg/m2, and paclitaxel at 175 mg/m2 (205 .mu.mol/m2) and Compound
(1) at a doses ranging among 44-525 mg/m2 (110-1311 .mu.mol/m2).
Table 1 shows the eight different doses #1-#8 in mg/m.sup.2 and
.mu.mol/m.sup.2.
TABLE-US-00001 TABLE 1 #1 #2 #3 #4 #5 #6 #7 #8 Compound (1), 44 44
88 175 263 350 438 525 mg/m.sup.2 Compound (1), 110 110 220 437 657
874 1094 1311 .mu.mol/m.sup.2 Paclitaxel, 135 175 175 175 175 175
175 175 mg/m.sup.2 Paclitaxel, 158 205 205 205 205 205 205 205
.mu.mol/m.sup.2
[0112] No serious effects specifically attributable to Compound (1)
were observed. Paclitaxel dose limiting toxicities occurred in a
single patient in each of the top three dose levels (neutropenia,
arthralgia, and febrile neutropenia with mucositis) resulting in
cohort expansion. Compound (1) exhibited linear PK that was
unaffected by paclitaxel dose, and was rapidly eliminated from
plasma with terminal-phase half life of 0.94.+-.0.23 hours (h) and
total body clearance of 28.+-.8 Liters/hour/meter.sup.2
(L/h/m.sup.2). Its apparent volume of distribution was comparable
to total body water (V.sub.SS23.+-.16 L/m.sup.2). Paclitaxel PK
appeared to be moderately dependent on the Compound (1) dose, as
indicated by a significant trend toward decreasing clearance, and
increase in peak plasma concentration and V.sub.SS, but without
affecting the terminal phase half-life. These observations are
consistent with competitive inhibition of paclitaxel hepatic
metabolism. Increased toxicity at higher dose levels was consistent
with a moderate increase in systemic exposure to paclitaxel.
Induction of Hsp70 protein in plasma was dose dependent, peaking
between about 8 hours to about 24 hours after dosing.
[0113] FIGS. 1A, 1B, and 1C are bar graphs showing the percent
increase in Hsp70 plasma levels associated with administration of
the Compound (1)/paclitaxel combination therapy at 1 hour (FIG.
1A), 5 hours (FIG. 1B), and 8 hours (FIG. 1C) after administration.
Significant rises in Hsp70 levels occurred for at least one patient
at the 88 mg/m2 (220 .mu.mol/m2) Compound (1) dose, where Hsp70
levels nearly doubled in a percent increase of about 90%. At the
175 mg/m2 (437 .mu.mol/m2) Compound (1) dose, Hsp70 concentrations
more than doubled in two patients; at the 263 mg/m2 (657
.mu.mol/m2) Compound (1) dose, Hsp70 concentrations roughly doubled
in two patients and increased by more than 250% in a third patient;
at the 350 mg/m2 (874 .mu.mol/m2) Compound (1) dose, Hsp70
concentrations increased more than 200% in all patients and
increased by as much as 500% in two patients; at the 438 mg/m2
(1094 .mu.mol/m2) Compound (1) dose, Hsp70 concentrations roughly
doubled in two patients, increased by over 2005 in one patient, and
increased by as much as 500% in another patient.
[0114] Time to progression will be measured as the time from
patient randomization to the time the patient is first recorded as
having tumor progression according to the RECIST (Response
Evaluation Criteria in Solid Tumors Group) criteria; see Therasse
P, Arbuck S G, Eisenhauer E A, Wanders J, Kaplan R S, Rubinstein L,
et al. New guidelines to evaluate the response to treatment in
solid tumors. J Natl Cancer Inst 2000; 92:205-16, the entire
teachings of which are incorporated by reference. Death from any
cause will be considered as progressed.
[0115] Time to progression can be performed on the randomized
sample as well as the efficacy sample. Treatment groups can be
compared using the log-rank test and Kaplan-Meier curves of time to
progression can be presented.
[0116] Thus, the combination of a bi(thio-hydrazide) amide and
taxane dramatically increased plasma Hsp70 levels in patients,
giving significant increases for patients at a combined paclitaxel
dose of 175 mg/m2 (205 .mu.mol/m2) and Compound (1) doses ranging
from 88 through 438 mg/m2 (220-1094 .mu.mol/m2). Moreover, the
combination was well-tolerated, with adverse events consistent with
those expected for paclitaxel alone.
Example 4
A 2 Stage Phase 2 Study Shows the Disclosed Combination Therapy is
Effective for Treating Advanced Metastatic Melanoma
[0117] The following study of Compound (1) and paclitaxel in
patients with advanced metastatic melanoma was initiated based on
the biological activity shown by the results of the above Phase I
study, where the combined administration Compound (1) and
paclitaxel led to dose-related Hsp70 induction.
[0118] The study included a Stage 1 initial safety assessment of
the weekly dose schedule, where Compound (1) 106 mg/m2 (265
.mu.mol/m2) and paclitaxel at 80 mg/m2 (94 .mu.mol/m2) were
administered weekly for 3 weeks out a 4 week period. The dose of
Compound (1) was then escalated to 213 mg/m2 (532 .mu.mol/m2) in
combination with the paclitaxel at 80 mg/m2 (94 .mu.mol/m2). The
higher tolerated dose level was expanded to a total of 20 patients
(Stage 1).
[0119] A total of 7 patients were treated in the initial safety
assessment, 3 at the lower dose level and 4 at the higher. In the
absence of dose-limiting toxicities in either group, the higher
dose level was chosen as the dose of interest and additional
patients were enrolled to complete stage 1. Adverse events seen
were as expected for paclitaxel chemotherapy administration. Of 20
evaluable patients, 11 were stable at 3 months for 55% NPR.
[0120] The study will continue to Stage 2 if 7 or more patients
have a response of stable disease or better, or at least 2 patients
have a partial response or better. A safety assessment was
performed with the first 6 patients enrolled a s the weekly dose
schedule had not previously been studied in humans. The primary
endpoint is non-progression rate (NPR) at 3 months and response
rate. Pharmacodynamic parameters include pre and post-dose NK cell
activity in blood and when possible, tumor biopsies.
[0121] Table 2 shows the significant preliminary results of
anticancer efficacy and NK cell activity results when assayed 7
days after the second dose for different subjects. The
Effector/Target data shows the ratio of the subjects PBMC cells to
the NK assay target cells. The pre and post dose column values show
the percent of tumor cells lysed before dosing with Paclitaxel and
Compound (1). Best Response indicates an evaluation of the
patient's tumor: SD indicates less than 20% of an increase and less
than 30% of a decrease in the sum of the longest diameters as
compared to baseline; and PD=at least a 20% increase in the sum of
the longest diameters as compared to baseline. NK Activity
indicates the change in NK activity before and after dosing.
[0122] Table 2 shows that for patients completing the study
(#12-#20, #22), three patients had less than 20% of an increase and
less than 30% of a decrease in the sum of the longest diameters as
compared to baseline, while seven patients had at least a 20%
increase in the sum of the longest diameters as compared to
baseline. For NK cell activity, four of the original patients
showed a statistically significant increase between pre- and
post-dose treatment.
TABLE-US-00002 TABLE 2 % tumor dosing information Effec- cell lysis
Cmpnd Best Response Sub- tor/ pre- post- Paclitaxel, (1) cycle 2 NK
ject Target dose dose mg/M.sup.2 mg/M.sup.2 week 4 activity 12 80:1
2.32 7.74 80 106 SD increase 13 80:1 6.13 2.43 80 106 PD decrease
14 80:1 3.83 10.77 80 213 SD increase 15 (40:1) 3.5 10.01 80 213 PD
(increase) 16 80:1 19.71 19.78 80 213 SD no change 17 80:1 41.61
26.52 80 213 PD decrease 18 80:1 8.6 8.64 80 213 PD no change 19
80:1 24.76 18.77 80 213 PD decrease 20 80:1 16.49 5.2 80 213 PD
decrease 21 80:1 15.4 26.31 80 213 NA increase 22 80:1 10.81 7.2 80
213 PD decrease
[0123] The combination therapy was well-tolerated on the weekly
schedule. Enrollment in the randomized portion will assess the
activity of Compound (1) in combination with paclitaxel versus
paclitaxel alone.
[0124] Stage 2 is planned to be a randomized 2-arm study comparing
the drug combination to paclitaxel alone. The criterion for
continuation to Stage 2 is >=50% non-progression rate (NPR) at
two months. A total of 78 patients are to be randomized 2:1
(combination:control). The primary endpoint is time to progression;
secondary endpoints are response rate, survival, and quality of
life. Pharmacodynamic parameters will include pre- and post-dose
measurements of NK cell activity in blood and, when possible, tumor
biopsies.
Example 5
A Phase 2 Study Shows the Disclosed Combination Therapy is
Effective for Treating Soft Tissue Sarcomas
[0125] The following study of Compound (1) and paclitaxel in
patients with soft tissue sarcomas was initiated based on the
biological activity shown by the results of the above Phase I
study, where the combined administration Compound (1) and
paclitaxel led to dose-related Hsp70 induction.
[0126] The study is a 2 stage design, enrolling 30 patients in the
first stage and adding 50 patients to total 80 if certain
continuation criteria are met. Major inclusion criteria are
refractory or recurrent soft tissue sarcomas other than
gastrointestinal stromal tumor (GIST), with evidence of recent
progression. Patients are treated weekly, 3 weeks out of every 4
week cycle with 213 mg/m2 Compound (1) and 80 mg/m2 paclitaxel. For
example, the compounds were administered together 3 weeks out of 4
on Days 1, 8, and 15 of a 28 day cycle as a 1 hour IV infusion. 30
Patients have been enrolled to completed accrual of Stage 1.
[0127] As used herein, "soft-tissue sarcomas" (STS) are cancers
that begin in the soft tissues that support, connect, and surround
various parts of the body for example, soft tissues such as
muscles, fat, tendons, nerves, and blood vessels, lymph nodes, or
the like. Such STSs can occur anywhere in the body, though
typically about one half occur in the limbs. In various
embodiments, STSs can include one or more cancers selected from
liposarcoma, fibrosarcoma, malignant fibrous histiocytoma
leiomyosarcoma, neurofibrosarcoma, rhabdomyosarcoma, synovial
sarcoma, or the like.
[0128] Table 3 shows the significant preliminary results of
anticancer efficacy and NK cell activity results when assayed 7
days after the second dose for different subjects. The
Effector/Target data shows the ratio of the subjects PBMC cells to
the NK assay target cells. The pre and post dose column values show
the percent of tumor cells lysed before dosing with Paclitaxel and
Compound (1). Best Response indicates an evaluation of the
patient's tumor: PR=at least a 30% decrease in the sum of the
longest diameters as compared to baseline; SD indicates less than
20% of an increase and less than 30% of a decrease in the sum of
the longest diameters as compared to baseline; and PD=at least a
20% increase in the sum of the longest diameters as compared to
baseline. NK Activity indicates the change in NK activity before
and after dosing.
[0129] Table 3 shows that for patients completing the study
(#23-#29, #31-33), five patients had less than 20% of an increase
and less than 30% of a decrease in the sum of the longest diameters
as compared to baseline, while five patients had at least a 20%
increase in the sum of the longest diameters as compared to
baseline. For NK cell activity, seven of the original patients
showed a statistically significant increase or no change between
pre- and post-dose treatment, while only four of the original
patients showed a decrease statistically significant increase
between pre- and post-dose treatment.
TABLE-US-00003 TABLE 3 % tumor cell dosing information Effec- lysis
Cmpnd Best Response Sub- tor/ pre- post- Paclitaxel, (1) NK ject
Target dose dose mg/M.sup.2 mg/M.sup.2 cycle 2 activity 23 80:1
4.28 30.48 80 213 PD increase 24 80:1 20.74 20.04 80 213 SD no
change 25 80:1 34.28 11.86 80 213 PD decrease 26 80:1 22.33 14.74
80 213 SD decrease 27 80:1 10.6 22.9 80 213 SD increase 28 80:1
17.93 28.13 80 213 SD increase 29 80:1 6.58 17.18 80 213 PD
increase 30 (40:1) 9.88 9.91 80 213 NA no change 31 80:1 2.62 5.46
80 213 SD increase 32 80:1 13.03 7.41 80 213 PD decrease 33 80:1
15.77 7.84 80 213 PD decrease
[0130] Patients are currently being evaluated through 3 months.
Adverse events seen were typical for paclitaxel administration on a
similar schedule. Assessment of NK activity is ongoing. The
addition of Compound (1) to the weekly paclitaxel schedule was
well-tolerated. Stage 1 accrual has completed, and patients are
currently being evaluated for the study continuation decision.
[0131] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims.
* * * * *