U.S. patent application number 13/030885 was filed with the patent office on 2012-02-23 for trp-p8 active compounds and therapeutic treatment methods.
This patent application is currently assigned to Genentech, Inc.. Invention is credited to Paul Polakis, Mark Reynolds.
Application Number | 20120045448 13/030885 |
Document ID | / |
Family ID | 33567725 |
Filed Date | 2012-02-23 |
United States Patent
Application |
20120045448 |
Kind Code |
A1 |
Reynolds; Mark ; et
al. |
February 23, 2012 |
TRP-P8 ACTIVE COMPOUNDS AND THERAPEUTIC TREATMENT METHODS
Abstract
Compounds of the disclosure provide compositions, which are
effective for prophylaxis and treatment of diseases or disorders,
such as cell-proliferation, angiogenesis, or apoptosis mediated
diseases. The disclosure encompasses compounds, analogs, prodrugs,
metabolites, and pharmaceutically acceptable salts thereof,
pharmaceutical compositions, and methods for prophylaxis and
treatment of diseases and other maladies or conditions involving
cancer, tumors, and like conditions. The disclosure also provides
therapeutic methods including the administration of an effective
amount of a compound of the disclosure.
Inventors: |
Reynolds; Mark; (Millbrae,
CA) ; Polakis; Paul; (Burlingame, CA) |
Assignee: |
Genentech, Inc.
South San Francisco
CA
|
Family ID: |
33567725 |
Appl. No.: |
13/030885 |
Filed: |
February 18, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10884379 |
Jul 2, 2004 |
7893072 |
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13030885 |
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60484526 |
Jul 2, 2003 |
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60491616 |
Jul 31, 2003 |
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Current U.S.
Class: |
424/158.1 ;
424/130.1; 435/375; 514/231.2; 514/617; 544/178; 564/123 |
Current CPC
Class: |
C07C 233/59 20130101;
A61K 31/513 20130101; A61K 31/00 20130101; A61P 35/00 20180101;
C07C 233/58 20130101; A61P 13/08 20180101; C07C 2602/08 20170501;
A61K 31/66 20130101; A61K 31/175 20130101; C07C 233/60 20130101;
C07D 295/096 20130101; C07D 239/22 20130101; A61P 9/14 20180101;
A61K 31/16 20130101; A61P 43/00 20180101; C07D 307/58 20130101;
C07D 295/135 20130101; C07C 2601/14 20170501 |
Class at
Publication: |
424/158.1 ;
514/231.2; 514/617; 544/178; 564/123; 424/130.1; 435/375 |
International
Class: |
A61K 39/395 20060101
A61K039/395; A61K 31/165 20060101 A61K031/165; C12N 5/02 20060101
C12N005/02; C07C 233/57 20060101 C07C233/57; A61P 35/00 20060101
A61P035/00; A61K 31/5375 20060101 A61K031/5375; C07D 265/30
20060101 C07D265/30 |
Claims
1. A compound of the formula IIa: ##STR00044## wherein R.sup.1 is
H, or (C.sub.1-C.sub.6)alkyl; R.sup.2 is phenyl or a substituted
phenyl of the formula (-PhR.sub.3R.sub.4R.sub.5R.sub.6R.sub.7)
##STR00045## where R.sub.3, R.sub.4, R.sub.6, and R.sub.7 are each
independently --H, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxyl, or halo; R.sub.5 is halo,
(C.sub.1-C.sub.6)alkyl, (C.sub.3-C.sub.12)cycloalkyl,
(C.sub.1-C.sub.6)alkoxyl, --C(.dbd.O)(C.sub.1-C.sub.6)alkyl or
(C.sub.1-C.sub.7)alkanoyl; or R.sub.5 is --NR.sub.8R.sub.9, where
R.sub.8 and R.sub.9 are each independently --H,
(C.sub.1-C.sub.6)alkyl, or R.sub.8 and R.sub.9 together with the
nitrogen to which they are attached form a morpholino, pyrrolidino,
piperidino, piperzino, indolino, benzimidazolino, azetidino,
aziridino, azepino, 1,4-oxazino, or thiomorpholino ring; or R.sub.4
and R.sub.5 together with the phenyl to which they are attached
form a ring having 4 to 7 atoms and the ring having from 1 to 3
unsaturations; and stereoisomeric forms, mixtures of stereoisomeric
forms; or a pharmaceutically acceptable salt thereof, provided that
when R.sub.3, R.sub.4, R.sub.6, and R.sub.7 of
-PhR.sub.3R.sub.4R.sub.5R.sub.6R.sub.7 are --H, R.sub.5 is other
than --CH.sub.3, --OCH.sub.3, --OH, --F, or --NO.sub.2; and
provided that R.sub.2 is other than 3-hydroxy-4-methyl-phenyl; and
further provided that R.sub.2 is other than 2-hydroxy-naphthyl, or
pyridyl.
2. The compound of claim 1, wherein the compound of the formula
(IIa) is: 2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-morpholin-4-yl-phenyl)-amide;
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(3-chloro-4-methoxy-phenyl)-amide;
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-sec-butyl-phenyl)-amide;
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-tert-butyl-phenyl)-amide;
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-propyl-phenyl)-amide; and
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-isopropyl-3-methyl-phenyl)-amide, or pharmaceutically acceptable
salts thereof.
3. A pharmaceutical composition comprising the compound of claim 1
and a pharmaceutically acceptable excipient.
4. The composition of claim 3, further comprising an effective
amount of at least one additional chemotherapeutic agent for
treating or preventing a disease or Trp-p8 receptor related
condition.
5. The composition of claim 4, wherein the chemotherapeutic agent
is an antibody that causes apoptosis, inhibits angiogenesis, or
both.
6. The composition of claim 5, wherein the chemotherapeutic agent
is an anti-VEGF antibody.
7. A method of treating cancer comprising contacting the cancer
cells comprising a Trp-p-8 receptor with an effective amount of a
compound of claim 1.
8. A method of treating cancer comprising contacting the cancer
cells comprising a Trp-p-8 receptor with an effective amount of a
pharmaceutical composition of claim 3.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of application Ser. No.
10/884,379, filed Jul. 2, 2004, which application claims the
benefit of provisional application Ser. No. 60/484,526, filed Jul.
2, 2003 and provisional application Ser. No. 60/491,616, filed Jul.
31, 2003, which applications are incorporated herein by reference
in their entirety.
BACKGROUND
[0002] Compounds which produce a physiological cool sensation when
applied to the skin are known, see for example, "New Compounds with
the Menthol Cooling Effect," H. R. Watson, et al., J. Soc. Cosmet.
Chem., 1978, 29, 185-200.
[0003] Wei, E. T., et al., J. Pharm. Pharmacol., 1983, 35(2),
110-112, describe a compound named "icilin" for its cool-sensation
producing properties, (also known as AG-3-5) or
3-(2-Hydroxy-phenyl)-6-(3-nitro-phenyl)-3,4-dihydro-1H-pyrimidin-2-one,
of the formula:
##STR00001##
[0004] Still other compounds having cooling action have been
recently reported, see H. Ottinger, et al., J. Agric. Food Chem.,
2001, 49, 5383-5390.
[0005] U.S. Pat. No. 4,150,052, discloses menthane carboxamide
compounds, for example, of the formula IIa3-1:
##STR00002##
that are reported to have a physiological cooling action on the
skin.
[0006] U.S. Pat. No. 4,070,496, discloses certain phosphine oxide
(R.sub.1R.sub.2R.sub.3P.dbd.O) compounds and compositions that are
reported to have a physiological cooling action on the skin.
[0007] U.S. Pat. No. 3,821,221, discloses certain
tetrahydropyrimidine-2-one compounds that are reported to have
central nervous system activity as depressants or stimulants.
[0008] Recently, certain TRP receptors have been shown to have a
role in thermosensation, see D. D. McKemy, et al., "Identification
of a Cold Receptor Reveals a General Role for TRP Channels in
Thermosensation," Nature, 2002, Mar. 7; 416(6876):52-8. For a
recent review of "The TRP Ion Channel Family," see D. E. Clapham,
et al., Nature Reviews, Neuroscience, 2001, 2, 387-396
<www.nature.com/reviews/neuro>. Okazawa et al Neuroscience
letters (2004 Apr. 8), 359(1-2):33-6; Nealen et al Journal of
neurophysiology (2003 July), 90(1):515-20; Thut et al.,
Neuroscience (2003), 119(4):1071-83.
[0009] The gene Trp-p8 was discovered by screening a
prostate-specific subtracted cDNA library. The predicted protein
has significant homology with the transient receptor potential
(Trp) family of Ca.sup.2+ channel proteins. Northern blot analysis
indicates Trp-p8 expression within normal human tissues is mostly
restricted to prostate epithelial cells. In situ hybridization
analysis shows that Trp-p8 mRNA expression was at moderate levels
in normal prostate tissue and appears to be elevated in prostate
cancer. Trp-p8 mRNA was also expressed in a number of non-prostatic
primary tumors of breast, colon, lung, and skin origin, whereas
transcripts encoding Trp-p8 were hardly detected or not detected in
the corresponding normal human tissues (Tsavaler, et al Cancer
Research (2001), 61(9):3760-3769).
[0010] Immunotherapy of prostate carcinoma (PCa) largely depends on
the identification of suitable target antigens that are present in
a high percentage of prostate tumors. The putative calcium channel
protein, Trp-p8, is associated with loss of Trp-p8 mRNA expression
and a significantly shorter time to PSA relapse-free survival. The
identification of Trp-p8 is associated with prostate cancer
outcome, and suggests an integral role for this receptor in
prostate carcinogensis. Immunogenic peptides derived from the
prostate-specific protein transient receptor potential-p8 (Trp-p8)
that is recognized by cytotoxic T lymphocytes from PCa patients
have been reported (Kiessling, et al (2003) Prostate 56(4):270-279;
Henshall, et al Cancer Research (2003), 63(14):4196-4203; Fuessel,
et al International Journal of Oncology (2003), 23(1):221-228; US
2003108963 A1). Identification of therapeutic agents effective in
the treatment of neoplastic, hyperplastic, and like diseases or
conditions continues to be the subject of significant research
efforts. Recent work indicates that certain therapeutic agents in
combination with certain antibody preparations can be effective in
treating angiogenic related disorders, and like diseases or
conditions, see for example, U.S. Pat. No. 6,582,959.
[0011] There is currently a need for therapeutic agents and methods
that are useful to treat diseases and conditions that are
associated with regulation of the Trp-p8 receptor. There is also a
need for therapeutic agents and treatment methods, which are
specific and selective toward cancerous cells and have low
cytotoxicity toward healthy cells.
[0012] There is also a need for therapeutic agents in combination
with additional chemotherapeutic agents, including, for example,
antibody preparations, and combination therapeutic treatment
methods thereof, that are useful to treat diseases and conditions
that are associated with regulation of the Trp-p8 receptor.
SUMMARY
[0013] It has now been discovered certain compounds, including some
known to produce a physiological cool sensation ("cool-genic"),
such as the above mentioned tetrahydropyrimidine-2-ones, phosphine
oxides, menthane carboxamide compounds, alkyl substituted alkyl
amide compounds, and alpha-keto enamines, exhibit useful biological
activity, such as anti-tumor activity.
[0014] Accordingly, the present disclosure provides, in exemplary
embodiments, compounds which activate the Trp-p8 receptor. In
embodiments, the present disclosure provides compounds and
treatment methods, which cause increased calcium ion flow into
cancerous cells (i.e., "flux activating" or "flux promoting"
compounds). In embodiments, the present disclosure provides
compounds, pharmaceutical compositions, and treatment methods,
which inhibit or kill cancerous cells, for example, by causing
apoptosis.
[0015] In embodiments, the present disclosure also provides:
[0016] a pharmaceutical composition comprising a compound of the
disclosure and a pharmaceutically acceptable excipient (the
composition preferably comprises a therapeutically effective amount
of the compound or salt);
[0017] a pharmaceutical composition for use in the treatment of
tumors, which comprises a compound of the disclosure, and
combinations thereof, or a pharmaceutically acceptable salt
thereof, and a pharmaceutically acceptable carrier.
[0018] a method for treating a disease or condition in a mammal
(e.g., a human) wherein a Trp-p8 receptor is implicated and
modulation of receptor function is desired comprising administering
an effective modulatory amount of a compound of the disclosure;
[0019] a method for treating or preventing a disease or Trp-p8
receptor related condition (e.g., tumors) in a mammal comprising
administering a therapeutically effective amount of a compound of
the disclosure;
[0020] a compound of the disclosure for use in medical diagnosis or
therapy (e.g., the treatment or prevention of Trp-p8 receptor
related disease or condition such as tumors);
[0021] the use of a compound of the disclosure to prepare a
medicament useful for treating or preventing a disease or Trp-p8
receptor related condition (e.g., the treatment or prevention of
Trp-p8 receptor related disease or condition such as tumors);
[0022] a method of treating cancer, for example, tumors, comprising
administering to a mammal in need of such treatment, an effective
amount of a compound of the disclosure;
[0023] a method for modulating Trp-p8 receptor function comprising
administering an effective modulatory amount of a compound of the
disclosure; and
[0024] a method for modulating Trp-p8 receptor function comprising
contacting an Trp-p8 receptor with an effective modulatory amount
of a compound of the disclosure.
[0025] We have also discovered certain compounds of the disclosure,
for example, those compounds known to produce a physiological cool
sensation ("cool-genic") and structurally related compounds, such
as the above mentioned tetrahydropyrimidine-2-ones, phosphine
oxides, menthane carboxamide compounds, alkyl substituted alkyl
amide compounds, alpha-keto enamines, and like compounds, when used
in combination with additional chemotherapeutic agents, including,
for example, antibodies, are capable of useful biological activity,
such as anti-tumor activity.
[0026] Accordingly, the present disclosure provides in embodiments,
therapeutic combinations of a compound of the disclosure (which can
activate the Trp-p8 receptor), and additional chemotherapeutic
agents, including, for example, an antibody. In embodiments, the
present disclosure provides such therapeutic combinations which can
be effective in the treatment of cancerous cells. In embodiments,
the present disclosure provides such therapeutic combinations and
treatment methods thereof, which can be effective in inhibiting or
killing cancerous cells, for example, by causing apoptosis,
inhibiting angiogenesis, or both.
[0027] In embodiments, the present disclosure also provides:
[0028] a composition comprising a compound of the disclosure in
combination with an antibody and a pharmaceutically acceptable
carrier;
[0029] a pharmaceutical composition comprising therapeutic
combinations of a compound of the disclosure and an antibody, and a
pharmaceutically acceptable excipient (the composition preferably
comprises a therapeutically effective amount of the compound or
salt and a therapeutically effective amount of at least one
additional chemotherapeutic agent, for example, an anti-angiogenic
antibody);
[0030] a pharmaceutical composition for use in the treatment of
tumors, which comprises an effective amount of a compound of the
disclosure, or a pharmaceutically acceptable salt thereof, in
combination with an effective anti-tumor amount of at least one
additional chemotherapeutic agent, for example, an anti-angiogenic
antibody, and a pharmaceutically acceptable carrier;
[0031] a method for treating a disease or condition in a mammal
(e.g., a human) wherein a Trp-p8 receptor is implicated (e.g.,
wherein the disease or condition is characterized by
over-expression Trp-p8 receptors) and modulation of receptor
function is desired comprising administering an effective
modulatory amount of a compound of the disclosure in combination
with an effective anti-cancer amount of at least one additional
chemotherapeutic agent, for example, an anti-angiogenic
antibody;
[0032] a method for treating or preventing a disease or Trp-p8
receptor related condition (e.g., tumors) in a mammal comprising
administering a therapeutically effective amount of a combination
of compound of the disclosure and at least one additional
chemotherapeutic agent, for example, an anti-angiogenic
antibody;
[0033] a compound of the disclosure in combination with at least
one additional chemotherapeutic agent, for example, an effective
amount of an anti-angiogenic antibody for use in medical diagnosis
or therapy (e.g., the treatment or prevention of Trp-p8 receptor
related disease or condition such as tumors);
[0034] the use of a compound of the disclosure in combination with
an effective amount of at least one additional chemotherapeutic
agent, for example, an anti-angiogenic antibody to prepare a
medicament useful for treating or preventing a disease or Trp-p8
receptor related condition (e.g., the treatment or prevention of
Trp-p8 receptor related disease or condition such as tumors);
[0035] a method of treating cancer, for example, tumors, comprising
administering to a mammal in need of such treatment, an effective
amount of a compound of the disclosure in combination with an
effective amount of at least one additional chemotherapeutic agent,
for example, an anti-angiogenic antibody;
[0036] a method for modulating Trp-p8 receptor function comprising
administering an effective modulatory amount of a compound of the
disclosure in combination with an effective amount of at least one
additional chemotherapeutic agent, for example, an anti-angiogenic
antibody; and
[0037] a method for modulating Trp-p8 receptor function comprising
contacting an Trp-p8 receptor with an effective modulatory amount
of a compound of the disclosure in combination with an effective
amount of at least one additional chemotherapeutic agent, for
example, an anti-angiogenic antibody.
[0038] We have also discovered certain other compounds, such as
certain menthane carboxamide compounds described below,
characterized in that they also exhibit useful biological activity,
such as cell killing anti-tumor activity.
[0039] Accordingly, in embodiments, the present disclosure also
provides compounds of the formula IIa:
##STR00003##
[0040] wherein
[0041] R.sub.1 is H, or (C.sub.1-C.sub.6)alkyl;
[0042] R.sub.2 is phenyl or a substituted phenyl of the formula
(-PhR.sub.3R.sub.4R.sub.5R.sub.6R.sub.7)
##STR00004##
[0043] where
[0044] R.sub.3, R.sub.4, R.sub.6, and R.sub.7 are each
independently --H, (C.sub.1-C.sub.6)alkyl,
(C.sub.1-C.sub.6)alkoxyl, or halo;
[0045] R.sub.5 is halo, (C.sub.1-C.sub.6)alkyl,
(C.sub.3-C.sub.12)cycloalkyl, (C.sub.1-C.sub.6)alkoxyl,
--C(.dbd.O)(C.sub.1-C.sub.6)alkyl or (C.sub.1-C.sub.7)alkanoyl;
[0046] or R.sub.5 is --NR.sub.8R.sub.9, where R.sub.8 and R.sub.9
are each independently --H, (C.sub.1-C.sub.6)alkyl, or R.sub.8 and
R.sub.9 together with the nitrogen to which they are attached form
a morpholino, pyrrolidino, piperidino, piperzino, indolino,
benzimidazolino, azetidino, aziridino, azepino, 1,4-oxazino, or
thiomorpholino ring;
[0047] or R.sub.4 and R.sub.5 together with the phenyl to which
they are attached form a ring having 4 to 7 atoms and the ring
having from 1 to 3 unsaturations; and
[0048] stereoisomeric forms, mixtures of stereoisomeric forms;
[0049] or a pharmaceutically acceptable salt thereof,
[0050] provided that when R.sub.3, R.sub.4, R.sub.6, and R.sub.7 of
-PhR.sub.3R.sub.4R.sub.5R.sub.6R.sub.7 are --H, R.sub.5 is other
than --CH.sub.3, --OCH.sub.3, --OH, --F, or --NO.sub.2; and
[0051] provided that R.sub.2 is other than
3-hydroxy-4-methyl-phenyl; and further provided
[0052] that R.sub.2 is other than 2-hydroxy-naphthyl, or
pyridyl.
[0053] In embodiments, the present disclosure also provides a
compound of the above mentioned formula IIa, which is characterized
in that the compound is effective in killing cells expressing
TRP-p8 but where the calcium ion flux of the expressing cell need
not be substantially changed by the presence of the compound.
[0054] In embodiments, the present disclosure also provides:
[0055] a method for killing target cells which express Trp-p8 but
without substantially changing the calcium flux characteristics of
the target cells (e.g., tumors) in a mammal comprising
administering a therapeutically effective amount of a compound of
the disclosure of the formula IIa;
[0056] a method for treating a disease or condition in a mammal
(e.g., a human) comprising administering an effective amount of a
compound of the disclosure of the formula IIa, which compound is
cytotoxic with respect to the Trp-p8 receptor and increased calcium
ion flux;
[0057] a method for treating or preventing a disease or Trp-p8
receptor related condition (e.g., tumors) in a mammal comprising
administering a therapeutically effective amount of a compound of
the disclosure of the formula IIa, which compound is cytotoxic with
respect to the Trp-p8 receptor; and
[0058] any of the above methods for killing cells, for treating a
disease or condition, or treating or preventing a disease
comprising administering a therapeutically effective amount of a
compound of the disclosure of the formula IIa in combination with
another compound of the disclosure, an anti-angiogenic antibody, or
mixtures thereof.
[0059] These and other embodiments are illustrated herein.
BRIEF DESCRIPTION OF THE DRAWINGS
[0060] FIG. 1A-D illustrate the effectiveness of selected
cool-genic compounds of the present disclosure in killing human
cells expressing Trp-p8 compared to the relative insensitivity of
human cells not expressing Trp-p8.
[0061] FIG. 2 illustrates the relative growth rates for cloned
cancerous cell lines (PC3/Trp-p8) which express Trp-p8 compared to
a control cell line (PC3-Neo) which does not express Trp-p8.
DETAILED DESCRIPTION
[0062] The present disclosure provides the abovementioned
pharmaceutical compositions and methods of treatment. The present
disclosure also provides the abovementioned compounds of the
disclosure of the formula IIa, pharmaceutical compositions
including a compound of the disclosure of the formula IIa, and
methods of treatment therewith, and which compounds, and
pharmaceutical compositions, are cytotoxic to Trp-p8 expressing
cells.
[0063] Examples of cool-genic compounds are, for example, a
compound of the formulas (I-XIII):
##STR00005##
wherein R.sub.1 and R.sub.2 are each independently H, alkyl, Het,
or aryl, or as disclosed in U.S. Pat. No. 3,821,221;
##STR00006##
wherein R.sub.1 and R.sub.2 are each independently H, alkyl, or
aryl, or as disclosed, for example, in U.S. Pat. No. 4,150,052, and
J. Soc. Cosmet. Chem., 1978, 29, 185-200;
##STR00007##
wherein R.sub.1, R.sub.2, and R.sub.3 are each independently H,
alkyl, or aryl, or as disclosed, for example, in J. Soc. Cosmet.
Chem., 1978, 29, 185-200 (and references cited therein such as
reference 1);
##STR00008##
[0064] wherein R.sub.1, R.sub.2, and R.sub.3 are each independently
linear or branched alkyl or cycloalkyl, or as disclosed, for
example, in U.S. Pat. No. 4,070,496;
##STR00009##
wherein R.sub.3 is --OH, --S(O)R.sub.1, --P(.dbd.O)R.sub.1R.sub.2,
--CO.sub.2H, --C(.dbd.O)NH.sub.2, --OC(.dbd.O)--CH(OH)--CH.sub.3,
--C(.dbd.O)OC.sub.nH.sub.2m--OH, where n is 1-4,
--NR.sub.1--C(.dbd.O)NR.sub.1R.sub.2, --SO.sub.2R.sub.1,
--SO.sub.2NR.sub.1R.sub.2, --SONR.sub.1R.sub.2, and where R.sub.1
and R.sub.2 are each independently H, alkyl, or aryl, and R.sub.4
is H, or R.sub.3 and R.sub.4 taken together with the carbon atom to
which they are attached is a 5-member ketal ring optionally having
an hydroxymethyl substituent of the formula
--OCH.sub.2--CH(CH.sub.2--OH)--O--, or as disclosed, for example,
in J. Soc. Cosmet. Chem., 1978, 29, 185-200;
[0065] a core selected from the (--X) substituted cyclic or
branched hydrocarbons of group VI:
##STR00010##
wherein X is an N-alkyl carboxamide, --C(.dbd.O)NR.sub.1R.sub.2,
where R.sub.1 and R.sub.2 are each independently H, alkyl, or aryl,
or R.sub.1 and R.sub.2 taken together with the nitrogen atom to
which they are attached is a 5- or 6-membered saturated or
unsaturated heterocyclic (Het) ring which is optionally substituted
with an oxygen (--O--) ring heteroatom, or as disclosed, for
example, in J. Soc. Cosmet. Chem., 1978, 29, 185-200;
##STR00011##
wherein X is an N-alkyl carboxamide, --C(.dbd.O)NHR.sub.1, where
R.sub.1 is alkyl or substituted alkyl, such as in --C(.dbd.O)NHEt
or --C(.dbd.O)NHCH.sub.2CO.sub.2Et, or alkyl sulfone, such as in
--SO.sub.2Et, or as disclosed, for example, in J. Soc. Cosmet.
Chem., 1978, 29, 185-200;
[0066] alkyl substituted urea compounds, for example, of the
formula VIII:
##STR00012##
or as disclosed, for example, in J. Soc. Cosmet. Chem., 1978, 29,
185-200;
##STR00013##
wherein R.sub.1 and R.sub.2 are each independently alkyl, or
R.sub.1 and R.sub.2 taken together with the nitrogen atom to which
they are attached is a 5- or 6-membered saturated or unsaturated
heterocyclic (Het) ring which is optionally substituted with an
oxygen (--O--) ring heteroatom, such as a morpholino-ring, and the
ring can be optionally substituted with an alpha-CO.sub.2H or an
alpha-CO.sub.2CH.sub.3 substituent, and R.sub.3, R.sub.4, and
R.sub.5 are each independently H or alkyl, or as disclosed, for
example, in J. Agric. Food Chem., 2001, 49, 5383-5390;
##STR00014##
wherein R.sub.1 and R.sub.2 are each independently alkyl, or
R.sub.1 and R.sub.2 taken together with the nitrogen atom to which
they are attached is a 5- or 6-membered saturated or unsaturated
heterocyclic (Het) ring, and R.sub.3, R.sub.4, R.sub.5, and R.sub.6
are each independently H or alkyl, or as disclosed, for example, in
J. Agric. Food Chem., 2001, 49, 5383-5390;
##STR00015##
wherein R.sub.1 and R.sub.2 are each independently alkyl, or
R.sub.1 and R.sub.2 taken together with the nitrogen atom to which
they are attached is a 5- or 6-membered saturated or unsaturated
heterocyclic (Het) ring, and R.sub.3 and R.sub.4 are each
independently H or alkyl, or as disclosed, for example, in J.
Agric. Food Chem., 2001, 49, 5383-5390;
##STR00016##
wherein R.sub.1 and R.sub.2 are each independently alkyl, or
R.sub.1 and R.sub.2 taken together form a 5- or 6-membered
saturated or unsaturated ring, and R.sub.3 and R.sub.4 are each
independently H or alkyl, or as disclosed, for example, in J.
Agric. Food Chem., 2001, 49, 5383-5390; or
##STR00017##
wherein R.sub.1 to R.sub.5 are each independently H, alkyl, or
aryl, R.sub.6 is --OH, --S(O)R.sub.8, --P(.dbd.O)R.sub.8R.sub.9,
--CO.sub.2H, --C(.dbd.O)NH.sub.2, --OC(.dbd.O)--CH(OH)--CH.sub.3,
--C(.dbd.O)OC--H.sub.2--OH, where n is 1-4,
--NR.sub.8--C(.dbd.O)NR.sub.8R.sub.9, --SO.sub.2R.sub.8,
--SO.sub.2NR.sub.8R.sub.9, or --SONR.sub.8R.sub.9, and where the
R.sub.8 and R.sub.9 of R.sub.6 are each independently H, alkyl, or
aryl, and R.sub.7 is H, or R.sub.6 and R.sub.7 taken together with
the carbon atom to which they are attached is a 5-member ketal
ring, having an hydroxymethyl substituent, of the formula
--OCH.sub.2--CH(CH.sub.2--OH)--O--; or a pharmaceutically
acceptable salt thereof.
[0067] The following definitions are used, unless otherwise
described: halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy,
etc., denote both straight and branched groups; but reference to an
individual radical such as "propyl" embraces only the straight
chain radical, a branched chain isomer such as "isopropyl" being
specifically referred to.
[0068] "Alkyl" is C.sub.1-C.sub.18 hydrocarbon containing normal,
secondary, tertiary or cyclic carbon atoms. Examples are methyl
(Me, --CH.sub.3), ethyl (Et, --CH.sub.2CH.sub.3), 1-propyl (n-Pr,
n-propyl, --CH.sub.2CH.sub.2CH.sub.3), 2-propyl (i-Pr, i-propyl,
--CH(CH.sub.3).sub.2), 1-butyl (n-Bu, n-butyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-methyl-1-propyl (1-Bu,
i-butyl, --CH.sub.2CH(CH.sub.3).sub.2), 2-butyl (s-Bu, s-butyl,
--CH(CH.sub.3)CH.sub.2CH.sub.3), 2-methyl-2-propyl (t-Bu, t-butyl,
--C(CH.sub.3).sub.3), 1-pentyl(n-pentyl,
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-pentyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.3), 3-pentyl
(--CH(CH.sub.2CH.sub.3).sub.2), 2-methyl-2-butyl
(--C(CH.sub.3).sub.2CH.sub.2CH.sub.3), 3-methyl-2-butyl
(--CH(CH.sub.3)CH(CH.sub.3).sub.2), 3-methyl-1-butyl
(--CH.sub.2CH.sub.2CH(CH.sub.3).sub.2), 2-methyl-1-butyl
(--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.3), 1-hexyl
(--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3), 2-hexyl
(--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2CH.sub.3).sub., 3-hexyl
(--CH(CH.sub.2CH.sub.3)(CH.sub.2CH.sub.2CH.sub.3)),
2-methyl-2-pentyl (--C(CH.sub.3).sub.2CH.sub.2CH.sub.2CH.sub.3),
3-methyl-2-pentyl (--CH(CH.sub.3)CH(CH.sub.3)CH.sub.2CH.sub.3),
4-methyl-2-pentyl (--CH(CH.sub.3)CH.sub.2CH(CH.sub.3).sub.2),
3-methyl-3-pentyl (--C(CH.sub.3)(CH.sub.2CH.sub.3).sub.2),
2-methyl-3-pentyl (--CH(CH.sub.2CH.sub.3)CH(CH.sub.3).sub.2),
2,3-dimethyl-2-butyl (--C(CH.sub.3).sub.2CH(CH.sub.3).sub.2),
3,3-dimethyl-2-butyl (--CH(CH.sub.3)C(CH.sub.3).sub.3.
[0069] "Aryl" denotes a phenyl radical or an ortho-fused bicyclic
carbocyclic radical having about nine to twenty ring atoms in which
at least one ring is aromatic. Aryl (Ar) can include substituted
aryls, such as a phenyl radical having from 1 to 5 substituents,
for example, alkyl, alkoxy, and like substituents, and which
substituents are consistent with the compounds and inclusive of the
substituents disclosed in the above mentioned patents or
publications for compounds of the formulas (I-XIII).
[0070] The term "antibody" herein is used in the broadest sense and
specifically covers intact monoclonal antibodies, polyclonal
antibodies, multi-specific antibodies (e.g., bispecific antibodies)
formed from at least two intact antibodies, and antibody fragments,
so long as they exhibit the desired biological activity. An
antibody is a protein generated by the immune system that is
capable of recognizing and binding to a specific antigen. Described
in terms of its structure, an antibody is a Y-shaped protein
consisting of four amino acid chains, two heavy and two light. In a
simplified model sufficient for this appeal, each antibody has
primarily two regions: a variable region and a constant region. The
variable region, located on the ends of the arms of the Y, binds to
and interacts with the target antigen. This variable region
includes a complementary determining region (CDR) that recognizes
and binds to a specific binding site on a particular antigen. The
constant region, located on the tail of the Y, is recognized by and
interacts with the immune system (Janeway, C., Travers, P.,
Walport, M., Shlomchik (2001) Immuno Biology, 5th Ed., Garland
Publishing, New York). A target antigen generally has numerous
binding sites, also called epitopes, recognized by CDRs on multiple
antibodies. Each antibody that specifically binds to a different
epitope has a different structure. Thus, one antigen may have more
than one corresponding antibody.
[0071] The term "monoclonal antibody" as used herein refers to an
antibody obtained from a population of substantially homogeneous
antibodies, i.e., the individual antibodies comprising the
population are identical except for possible naturally occurring
mutations that may be present in minor amounts. Monoclonal
antibodies are highly specific, being directed against a single
antigenic site. Furthermore, in contrast to polyclonal antibody
preparations which include different antibodies directed against
different determinants (epitopes), each monoclonal antibody is
directed against a single determinant on the antigen. In addition
to their specificity, the monoclonal antibodies are advantageous in
that they may be synthesized uncontaminated by other antibodies.
The modifier "monoclonal" indicates the character of the antibody
as being obtained from a substantially homogeneous population of
antibodies, and is not to be construed as requiring production of
the antibody by any particular method. For example, the monoclonal
antibodies to be used in accordance with the present invention may
be made by the hybridoma method first described by Kohler et al
(1975) Nature 256:495, or may be made by recombinant DNA methods
(see, U.S. Pat. No. 4,816,567). The "monoclonal antibodies" may
also be isolated from phage antibody libraries using the techniques
described in Clackson et al (1991) Nature, 352:624-628 and Marks et
al (1991) J. Mol. Biol., 222:581-597, for example.
[0072] The monoclonal antibodies herein specifically include
"chimeric" antibodies in which a portion of the heavy and/or light
chain is identical with or homologous to corresponding sequences in
antibodies derived from a particular species or belonging to a
particular antibody class or subclass, while the remainder of the
chain(s) is identical with or homologous to corresponding sequences
in antibodies derived from another species or belonging to another
antibody class or subclass, as well as fragments of such
antibodies, so long as they exhibit the desired biological activity
(U.S. Pat. No. 4,816,567; and Morrison et al (1984) Proc. Natl.
Acad. Sci. USA, 81:6851-6855). Chimeric antibodies of interest
herein include "primatized" antibodies comprising variable domain
antigen-binding sequences derived from a non-human primate (e.g.,
Old World Monkey, Ape etc) and human constant region sequences.
[0073] A "chemotherapeutic agent" is a chemical compound useful in
the treatment of cancer. Examples of chemotherapeutic agents
include alkylating agents such as thiotepa and CYTOXAN.RTM.
cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan
and piposulfan; aziridines such as benzodopa, carboquone,
meturedopa, and uredopa; ethylenimines and methylamelamines
including altretamine, triethylenemelamine,
triethylenephosphoramide, triethylenethiophosphoramide and
trimethylolomelamine; acetogenins (especially bullatacin and
bullatacinone); delta-9-tetrahydrocannabinol (dronabinol,
MARINOL.RTM.); beta-lapachone; lapachol; colchicines; betulinic
acid; a camptothecin (including the synthetic analogue topotecan
(HYCAMTIN.RTM.), CPT-11 (irinotecan, CAMPTOSAR.RTM.),
acetylcamptothecin, scopolectin, and 9-aminocamptothecin);
bryostatin; callystatin; CC-1065 (including its adozelesin,
carzelesin and bizelesin synthetic analogues); podophyllotoxin;
podophyllinic acid; teniposide; cryptophycins (particularly
cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin
(including the synthetic analogues, KW-2189 and CB1-TM1);
eleutherobin; pancratistatin; a sarcodictyin; spongistatin;
nitrogen mustards such as chlorambucil, chlornaphazine,
cholophosphamide, estramustine, ifosfamide, mechlorethamine,
mechlorethamine oxide hydrochloride, melphalan, novembichin,
phenesterine, prednimustine, trofosfamide, uracil mustard;
nitrosureas such as carmustine, chlorozotocin, fotemustine,
lomustine, nimustine, and ranimnustine; antibiotics such as the
enediyne antibiotics (e.g., calicheamicin, especially calicheamicin
gamma1I and calicheamicin omegaI1 (see, e.g., Agnew, Chem. Intl.
Ed. Engl., 33: 183-186 (1994)); dynemicin, including dynemicin A;
an esperamicin; as well as neocarzinostatin chromophore and related
chromoprotein enediyne antiobiotic chromophores), aclacinomysins,
actinomycin, authramycin, azaserine, bleomycins, cactinomycin,
carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin,
daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine,
ADRIAMYCIN.RTM. doxorubicin (including morpholino-doxorubicin,
cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and
deoxydoxorubicin), epirubicin, esorubicin, idarubicin,
marcellomycin, mitomycins such as mitomycin C, mycophenolic acid,
nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin,
quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,
ubenimex, zinostatin, zorubicin; anti-metabolites such as
methotrexate and 5-fluorouracil (5-FU); folic acid analogues such
as denopterin, methotrexate, pteropterin, trimetrexate; purine
analogs such as fludarabine, 6-mercaptopurine, thiamiprine,
thioguanine; pyrimidine analogs such as ancitabine, azacitidine,
6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine,
enocitabine, floxuridine; androgens such as calusterone,
dromostanolone propionate, epitiostanol, mepitiostane,
testolactone; anti-adrenals such as aminoglutethimide, mitotane,
trilostane; folic acid replenisher such as frolinic acid;
aceglatone; aldophosphamide glycoside; aminolevulinic acid;
eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate;
defofamine; demecolcine; diaziquone; elformithine; elliptinium
acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea;
lentinan; lonidainine; maytansinoids such as maytansine and
ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine;
pentostatin; phenamet; pirarubicin; losoxantrone; 2-ethylhydrazide;
procarbazine; PSK.RTM. polysaccharide complex (JHS Natural
Products, Eugene, Oreg.); razoxane; rhizoxin; sizofuran;
spirogermanium; tenuazonic acid; triaziquone;
2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2
toxin, verracurin A, roridin A and anguidine); urethan; vindesine
(ELDISINE.RTM., FILDESIN.RTM.); dacarbazine; mannomustine;
mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside
("Ara-C"); thiotepa; taxoids, e.g., TAXOL.RTM. paclitaxel
(Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE.TM.
Cremophor-free, albumin-engineered nanoparticle formulation of
paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.),
and TAXOTERE.RTM. doxetaxel (Rhone-Poulenc Rorer, Antony, France);
chloranbucil; gemcitabine (GEMZAR.RTM.); 6-thioguanine;
mercaptopurine; methotrexate; platinum analogs such as cisplatin
and carboplatin; vinblastine (VELBAN.RTM.); platinum; etoposide
(VP-16); ifosfamide; mitoxantrone; vincristine (ONCOVIN.RTM.);
oxaliplatin; leucovovin; vinorelbine (NAVELBINE.RTM.); novantrone;
edatrexate; daunomycin; aminopterin; ibandronate; topoisomerase
inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoids such
as retinoic acid; capecitabine (XELODA.RTM.); pharmaceutically
acceptable salts, acids or derivatives of any of the above; as well
as combinations of two or more of the above such as CHOP, an
abbreviation for a combined therapy of cyclophosphamide,
doxorubicin, vincristine, and prednisolone, and FOLFOX, an
abbreviation for a treatment regimen with oxaliplatin
(ELOXATIN.TM.) combined with 5-FU and leucovovin.
[0074] Also included in this definition are anti-hormonal agents
that act to regulate, reduce, block, or inhibit the effects of
hormones that can promote the growth of cancer, and are often in
the form of systemic or whole-body treatment. They may be hormones
themselves. Examples include anti-estrogens and selective estrogen
receptor modulators (SERMs), including, for example, tamoxifen
(including NOLVADEX.RTM. tamoxifen), EVISTA.RTM. raloxifene,
droloxifene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018,
onapristone, and FARESTON.RTM. toremifene; anti-progesterones;
estrogen receptor down-regulators (ERDs); agents that function to
suppress or shut down the ovaries, for example, leutinizing
hormone-releasing hormone (LHRH) agonists such as LUPRON.RTM. and
ELIGARD.RTM. leuprolide acetate, goserelin acetate, buserelin
acetate and tripterelin; other anti-androgens such as flutamide,
nilutamide and bicalutamide; and aromatase inhibitors that inhibit
the enzyme aromatase, which regulates estrogen production in the
adrenal glands, such as, for example, 4(5)-imidazoles,
aminoglutethimide, MEGASE.RTM. megestrol acetate, AROMASIN.RTM.
exemestane, formestanie, fadrozole, RIVISOR.RTM. vorozole,
FEMARA.RTM. letrozole, and ARIMIDEX.RTM. anastrozole. In addition,
such definition of chemotherapeutic agents includes bisphosphonates
such as clodronate (for example, BONEFOS.RTM. or OSTAC.RTM.),
DIDROCAL.RTM. etidronate, NE-58095, ZOMETA.RTM. zoledronic
acid/zoledronate, FOSAMAX.RTM. alendronate, AREDIA.RTM.
pamidronate, SKELID.RTM. tiludronate, or ACTONEL.RTM. risedronate;
as well as troxacitabine (a 1,3-dioxolane nucleoside cytosine
analog); antisense oligonucleotides, particularly those that
inhibit expression of genes in signaling pathways implicated in
abherant cell proliferation, such as, for example, PKC-alpha, Raf,
H-Ras, and epidermal growth factor receptor (EGF-R); vaccines such
as THERATOPE.RTM. vaccine and gene therapy vaccines, for example,
ALLOVECTIN.RTM. vaccine, LEUVECTIN.RTM. vaccine, and VAXID.RTM.
vaccine; LURTOTECAN.RTM. topoisomerase 1 inhibitor; ABARELIX.RTM.
rmRH; lapatinib ditosylate (an ErbB-2 and EGFR dual tyrosine kinase
small-molecule inhibitor also known as GW572016); antibodies that
have an anti-cancer effect, particularly those that bind to VEGF-1,
VEGF-2, VEGF-3, EGF-R, HER-2, CD20, and the like, and
pharmaceutically acceptable salts, acids or derivatives of any of
the above.
[0075] "Het" is a four- (4), five- (5), six- (6), or seven- (7)
membered saturated or unsaturated heterocyclic ring having 1, 2, 3,
or 4 heteroatoms selected from the group consisting of oxy, thio,
sulfinyl, sulfonyl, and nitrogen, which ring is optionally fused to
a benzene ring. Het includes "heteroaryl," which encompasses a
radical attached via a ring carbon of a monocyclic aromatic ring
containing five or six ring atoms consisting of carbon and 1, 2, 3,
or 4 heteroatoms each selected from the group consisting of
non-peroxide oxy, thio, and N(X) wherein X is absent or is H, O,
C.sub.1-4alkyl, phenyl or benzyl, as well as a radical of an
ortho-fused bicyclic heterocycle of about eight to ten ring atoms
derived therefrom, particularly a benz-derivative or one derived by
fusing a propylene, trimethylene, or tetramethylene diradical
thereto.
[0076] "Treat," "treatment," "treating," and like terms refer, in
embodiments, to lessen, to eliminate, to inhibit, to improve, to
alter, or to prevent a disease or condition, for example by
administration of an effective amount of a compound of the present
disclosure, or by administration of an effective amount of a
compound of the disclosure in combination with an effective amount
of an additional chemotherapeutic agent an anti-angiogenic
antibody, and can refer to curative therapy, prophylactic therapy,
and preventative therapy.
[0077] "Modulate," "modulation," "modulating," "modulatory," and
like terms refer to, in embodiments, the ability of an effective
amount of a compound of the present disclosure to, selectively and
at relatively low dose levels, adjust or alter the multi-valent ion
(such as calcium ions) permeability of cancerous cells, such as
those cells expressing Trp-p8, and in contrast to the relative
insensitivity of other cells, for example, healthy or non-cancerous
cells and cells not expressing Trp-p8.
[0078] "Cancer treatment," "treating cancer," and like terms, for
purposes of this disclosure refer, in embodiments, to a method of
treating cancer which includes contacting cancer cells with a
compound of the disclosure in order to achieve an inhibition of
cancer cell growth, a killing of cancer cells, increased patient
survival time, or a combination thereof, or contacting cancer cells
with a compound of the disclosure in combination with an
anti-angiogenic antibody in order to achieve an inhibition of
cancer cell growth, a killing of cancer cells, increased patient
survival time, an anti-angiogenic effect, or a combination thereof.
Treatment of cancer, by the method of the disclosure, also includes
contacting the cells with a compound of the disclosure to activate
Trp-p8 receptors in cancerous cells, such as tumors, to cause
elevated flux levels of multi-valent ions into the cells to cause
cell stasis or cell death. Cancer can include diseases in which
abnormal cells divide without control. Cancer cells can also invade
nearby tissue and can spread through the bloodstream and lymphatic
system to other parts of the body. The major categories of cancers
are carcinomas, sarcomas, leukemias, and lymphomas. Within these
major categories are numerous subgroups that generally describe the
organ in which the cancer originates, such as adenocarcinoma of the
stomach or oat cell carcinoma of the lung.
[0079] "Tumor" refers to, for example, an abnormal benign or
malignant mass of tissue that may not be inflammatory, arises from
cells of pre-existent tissue, and may possesses no physiological
function. Benign tumors, include for example, cysts, warts, moles,
and polyps, and generally do not spread to other parts of the body.
Malignant tumors are typically composed of cells that grow rapidly,
have other abnormal properties that distinguish them from normal
cells, and often invade other normal tissues.
[0080] "Vascular endothelial cell growth factor," or "VEGF," refers
to a mammalian growth factor as defined, for example, in U.S. Pat.
No. 5,332,671. The biological activity of native VEGF is shared by
any analog or variant thereof that promotes selective growth of
vascular endothelial cells but not of bovine corneal endothelial
cells, lens epithelial cells, adrenal cortex cells, BHK-21
fibroblasts, or keratinocytes.
[0081] "Angiogenic disorder" or "angiogenic defect" refers to an
abnormal condition that requires treatment with an agent that
inhibits angiogenesis, e.g., an angiostatic compound or composition
such as a combination of a compound of the disclosure and an
anti-angiogenic antibody. Such disorders include, for example,
types of cancer such as vascular tumors, e.g., hemangioma
(capillary and cavernous), glomus tumors, telangiectasia, bacillary
angiomatosis, hemangioendothelioma, angiosarcoma,
haemangiopericytoma, Kaposi's sarcoma, lymphangioma, and
lymphangiosarcoma, and tumor angiogenesis.
[0082] Administration "in combination with" one or more further
therapeutic agents includes simultaneous (concurrent) and
consecutive administration in any order.
[0083] The cool-genic compounds are suitable for use in mammals. As
used herein, "mammals" means any class of higher vertebrates that
nourish their young with milk secreted by mammary glands,
including, for example, humans, horses, cows, pigs, sheep, dogs,
rabbits, and monkeys.
[0084] "Apoptotic cell death," "programmed cell death," "apoptosis"
and like terms refer to any cell death that may result from the
complex cascade of cellular events that occur at specific stages of
cellular differentiation and in response to specific stimuli.
Apoptotic cell death can be characterized by condensation of the
cytoplasm and nucleus of dying cells. Apoptosis is an active
process requiring new protein synthesis. Typically, the process
requires ATP, involves new RNA and protein synthesis, and
culminates in the activation of endogenous endonucleases that
degrade the DNA of the cell, thereby destroying the genetic
template required for cellular homeostasis. Apoptosis is observed
in controlled deletion of cells during metamorphosis,
differentiation, and general cell turnover, and appears normally to
be regulated by receptor-coupled events. For these reasons,
apoptosis has been called "programmed cell death" or "cell
suicide." While every cell likely has a genetic program to commit
suicide, it is usually suppressed. Under normal circumstances, only
those cells no longer required by the organism activate this
self-destruction program.
[0085] "Therapeutically effective amount" means, in embodiments, a
dose of a compound of the disclosure, or a dose of a combination of
a compound of the disclosure and another chemotherapeutic agent,
with or without an excipient, that inhibits, reduces or eliminates
tumor growth, in vivo, in vitro, or both, by for example,
activating Trp-p8, stimulating apoptosis, or both. The exact dose
will depend on the purpose of the treatment, and will be
ascertainable by one skilled in the art using known techniques.
[0086] In one embodiment of the present disclosure, the compounds
of the disclosure are used in combination therapy, for example,
with other antibody therapeutic agents. In an embodiment, compounds
of the present disclosure are used in combination with known cancer
treating antibodies. See generally, for example: PCT/US02/19592;
PCT/US01/20118; PCT/US01/25464; PCT/US01/26626; PCT/US02/28859;
PCT/US02/41798; PCT/US02/12206; PCT/US03/11148; PCT/US02/12619; and
PCT/US02/33050. In another embodiment, compounds of the disclosure
are used in combination with an anti-VEGF antibody and like
antibodies including human, non-human, murine, hybrid, and chimeric
forms. See for example U.S. Pat. No. 6,582,959 (VEGF) and U.S.
Patent Application No. 2002/0122797 A1 (human VEGF).
[0087] In embodiments of the present disclosure, compounds of the
disclosure can be used in combination with other therapeutic
agents, such as the antibodies noted above, to treat immunological
diseases or conditions, for example, involving immune responsive
cells such as B-cells (B lymphocytes), T-cells (T lymphocytes),
accessory cells (macrophages and other antigen-presenting cells),
killer cells (NK and K cells), mast cells, and like cells.
[0088] In a preferred embodiment, compounds useful in the present
disclosure include a non-radio labeled compound for treatment of
non-central nervous system cells or diseases. In embodiments,
compounds of the present disclosure do not contain a radio-label
and are not radio-active. The unlabeled compounds of the present
disclosure can be used to kill cancer cells as illustrated herein,
for example, cells expressing Trp-p8, such as prostate cancer cells
and liver cancer cells.
[0089] A "subject" for the purposes of the present disclosure
includes both humans and other animals, particularly mammals. Thus,
the methods are applicable to both human therapy and veterinary
applications. In a preferred embodiment the subject is a mammal,
and in the most preferred embodiment the subject is human.
[0090] "Improved therapeutic outcome" or "decrease in the number of
tumor cells" or "decreased tumor size" means a 50% decrease,
preferably an 80% decrease, more preferably a 90% decrease, and
even more preferably a 100% decrease in the tumor size or volume, a
decrease in the number of detectable circulating cancer cells in
the blood, affected tissue, or organ as determined by examination
of a patient, samples taken from a patient prior to and following
treatment, or both.
[0091] "Compound," "molecule," "polypeptide," and like terms
include synthetically prepared compounds, genetically engineered
compounds (e.g., recombinant DNA expressed proteins), naturally
occurring compounds, and those produced in vivo after
administration of a different compound. The in vivo effects of
administered compounds described herein may not be exerted by those
compounds as such, but by one or more degradation products, such as
a metabolite, conjugate, clathrate, ion complex, chelate, hydrate,
solvate, or like biological transformations or combinations, of the
administered compound(s) or molecule(s). "Pharmaceutically
acceptable salts" can also include, in addition to those
illustrated herein, a subclass of salts present or formed in
vivo.
[0092] The present disclosure provides compounds that bind to
certain receptors in the TRP (transient receptor potential) ion
channel family. More particularly the present disclosure provides
compounds that specifically bind to the subgroup of long TRP (or
TRPM) channels, and most particularly to compounds that
specifically bind to the TRP channel called "Trp-p8" (or TRP-M8).
The Trp-p8 receptors are typically present at elevated levels in
cancers, such as prostate cancer. The compounds of the disclosure
are endowed with Trp-p8 receptor activating activity. Activation of
the Trp-p8 receptor causes increased calcium ion flow into
cancerous cells and ultimately cell death. Compounds of the present
disclosure are useful for, but not limited to, the treatment of
cell proliferation diseases or disorders, and stimulating
apoptosis. It is also well known in the art and as illustrated
herein how to determine Trp-p8 receptor activity, for example,
using the standard tests described herein, or using other similar
tests.
[0093] It will be appreciated by those skilled in the art that
compounds of the disclosure having a chiral center may exist in and
be isolated in optically active and racemic forms. Some compounds
may exhibit polymorphism. It is to be understood that the present
disclosure encompasses any racemic, optically-active, polymorphic,
tautomeric, or stereoisomeric form, or mixture thereof, of a
compound of the disclosure, which possesses the useful properties
described herein, it being well known in the art how to prepare
optically active forms (for example, by resolution of the racemic
form by recrystallization techniques, by synthesis from
optically-active starting materials, by chiral synthesis, or by
chromatographic separation using a chiral stationary phase). In
particular, it is understood that compounds of the disclosure, such
as of formulas (I-XIII), can contain chiral centers, for example,
in the R.sub.1 substituents of formula (I), the R.sub.3
substituents of formula (V), and in the R.sub.1 to R.sub.6
substituents of formula (XIII). It is also understood that
compounds of the disclosure, such as formula (I), can exist in the
"enol" form or the corresponding tautomeric "keto" form, and that
all such tautomers are included as compounds within the scope of
the present disclosure.
[0094] The carbon atom content of various hydrocarbon-containing
moieties is indicated by a prefix designating a lower and upper
number of carbon atoms in the moiety, i.e., the prefix C.sub.i-j
indicates a moiety of the integer "i" to the integer "j" carbon
atoms, inclusive. Thus, for example, C.sub.1-6alkyl or
(C.sub.1-C.sub.6)alkyl refers to alkyl of one to six carbon atoms,
inclusive.
[0095] The compounds of the present disclosure are generally named
according to the IUPAC nomenclature system. Abbreviations, which
are well known to one of ordinary skill in the art, may be used
(e.g., "Ph" for phenyl, "Me" for methyl, "Et" for ethyl, "h" for
hour or hours and "rt" for room temperature).
[0096] Specific and preferred values listed below for radicals,
substituents, and ranges, are for illustration only; they do not
exclude other defined values or other values within defined ranges
for the radicals and substituents. The compounds of the disclosure
include compounds of formula (I-XIII) having any combination of the
values, specific values, more specific values, and preferred values
described herein.
[0097] Specifically, aryl can be phenyl, naphthyl, anthracenyl,
phenanthrenyl, fluorenyl, tetrahydronaphthyl, or indanyl.
[0098] Specifically, alkyl such as (C.sub.1-6)alkyl can be methyl,
ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, tert-butyl,
pentyl, 3-pentyl, hexyl, or heptyl; (C.sub.2-C.sub.6)alkyl, can be
ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, tert-butyl,
pentyl, 3-pentyl, or hexyl; (C.sub.3-12)cycloalkyl can be
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,
cyclooctyl, bicyclic, or multi-cyclic substituents, such as of the
formulas
##STR00018##
[0099] C.sub.1-6alkoxy can be methoxy, ethoxy, propoxy, isopropoxy,
butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy;
--C(.dbd.O)alkyl or (C.sub.2--C.sub.7)alkanoyl can be acetyl,
propanoyl, butanoyl, pentanoyl, 4-methylpentanoyl, hexanoyl, or
heptanoyl; aryl can be phenyl, indenyl, or naphthyl; Het can be
pyrrolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, or
heteroaryl; and heteroaryl can be furyl, imidazolyl, triazolyl,
triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl,
pyrrolyl, pyrazinyl, tetrazolyl, pyridyl (or its N-oxide), thienyl,
pyrimidinyl (or its N-oxide), indolyl, isoquinolyl (or its N-oxide)
or quinolyl (or its N-oxide).
[0100] A specific value for Het is a five- (5), six- (6), or seven-
(7) membered saturated or unsaturated ring containing 1, 2, 3, or 4
heteroatoms, for example, non-peroxide oxy, thio, sulfinyl,
sulfonyl, and nitrogen; as well as a radical of an ortho-fused
bicyclic heterocycle of about eight to twelve ring atoms derived
therefrom, particularly a benz-derivative or one derived by fusing
a propylene, trimethylene, tetramethylene or another monocyclic Het
diradical thereto.
[0101] A specific compound of formula (I) is a compound of the
Formula (A)
##STR00019##
[0102] A specific compound of formula (II) is a compound of the
Formula (B)
##STR00020##
[0103] Another specific compound of formula (II) is a compound of
the Formula (C) showing a preferred stereochemistry:
##STR00021##
[0104] Another specific compound of formula (II) is a compound of
the Formula (D):
##STR00022##
[0105] A specific compound of formula (III) is a compound of the
Formula (E):
##STR00023##
[0106] A specific compound of formula (IV) is a compound of the
Formula (F):
##STR00024##
[0107] A specific compound of formula (V) is a compound of the
Formula (G):
##STR00025##
[0108] A specific compound of formula (VI) is a compound of the
Formula (H):
##STR00026##
[0109] A specific compound of formula (VII) is a compound of the
Formula (I'):
##STR00027##
[0110] A specific compound of formula (IX) is a compound of the
Formula (J):
##STR00028##
[0111] A specific compound of formula (X) is a compound of the
Formula (K):
##STR00029##
[0112] A specific compound of formula (XI) is a compound of the
Formula (L):
##STR00030##
[0113] A specific compound of formula (XII) is a compound of the
Formula (M):
##STR00031##
[0114] A specific compound of formula (XIII) is a compound of the
Formula (N):
##STR00032##
[0115] It is understood that the abovementioned specific compounds
of the Formulas (A-N), and all other compounds of the disclosure,
can be or include a pharmaceutically acceptable salt thereof.
[0116] A specific compound is
3-(2-Hydroxy-phenyl)-6-(3-nitro-phenyl)-3,4-dihydro-1H-pyrimidin-2-one;
or a pharmaceutically acceptable salt thereof.
[0117] Another specific compound is
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-methoxy-phenyl)-amide; or a pharmaceutically acceptable salt
thereof.
[0118] Another specific compound is 2-Isopropyl-2,3,
--N-trimethyl-butyramide; or a pharmaceutically acceptable salt
thereof.
[0119] Another specific compound is
1-(sec-Butyl-isobutyl-phosphinoyl)-heptane; or a pharmaceutically
acceptable salt thereof.
[0120] Another specific compound is
2-Isopropyl-5-methyl-cyclohexanol; or a pharmaceutically acceptable
salt thereof
[0121] A specific compound of formula IIa is a compound of the
formula Ha1:
##STR00033##
or a pharmaceutically acceptable salt thereof; wherein R.sub.1,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, and R.sub.7 are as defined
herein.
[0122] Other specific compounds of formula IIa are individual
compounds of the formulas IIa2-IIa12, or a pharmaceutically
acceptable salt thereof; wherein the R substituents are as defined
herein:
##STR00034## ##STR00035##
[0123] Specific compounds of formula IIa include: [0124]
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-morpholin-4-yl-phenyl)-amide, or a pharmaceutically acceptable
salt thereof; [0125] 2-Isopropyl-5-methyl-cyclohexanecarboxylic
acid (3-chloro-4-methoxy-phenyl)-amide, or a pharmaceutically
acceptable salt thereof; [0126]
2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-sec-butyl-phenyl)-amide, [0127] or a pharmaceutically acceptable
salt thereof; [0128] 2-Isopropyl-5-methyl-cyclohexanecarboxylic
acid indan-5-ylamide, or a pharmaceutically acceptable salt
thereof; [0129] 2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-tert-butyl-phenyl)-amide, or a pharmaceutically acceptable salt
thereof; [0130] 2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-propyl-phenyl)-amide, or a pharmaceutically acceptable salt
thereof and 2-Isopropyl-5-methyl-cyclohexanecarboxylic acid
(4-isopropyl-3-methyl-phenyl)-amide,
[0131] or a pharmaceutically acceptable salt thereof.
[0132] Preparative procedures, characterization, cool-genic
properties, odor properties, structure-coolgenic activity
relationships, design rules, and like information for compounds of
the formulas (I-XIII) and for the above mentioned specific
compounds of formulas A-N are reported in the corresponding above
mentioned publications or patents.
[0133] Compounds of the disclosure, such as compounds of formulas
B, C, D or N can be prepared as illustrated, for example, in the
scheme below, by procedures analogous thereto, by procedures in the
above mentioned publications or patents, or by procedures which are
known or would be readily evident to one of ordinary skill in the
art. All of the variables used in the schemes are as defined below
or elsewhere herein. Scheme 1 illustrates the preparation of
representative compounds of the disclosure, such as amide compound
3 (IIa-4-1). Chloro compound 1 was carboxylated via a Grignard
intermediate to afford carboxylic acid 2 and the acid 2 was
converted to the amide 3 and as described in Example 1 herein.
##STR00036##
[0134] An alternative preparative procedure for preparing compound
3 (IIa-4-1) and related amide compounds uses the corresponding acid
chloride of the above mentioned carboxylic acid compound 2. The
acid chloride of carboxylic acid compound 2 (readily prepared by
reaction with SOCl.sub.2 and like reagents) can be reacted with a
variety of primary or secondary amine compounds to form the
corresponding amides analogous to amide 3. Other compounds related
to amide 3 or formula II, were similarly prepared and as
illustrated and described herein.
[0135] For a synthetic procedure for making menthanecarboxylic acid
compound 2 (a starting material for IIa-4-1), see D. G. Rowsell,
Wilkinson Sword Ltd., U.K. Patent (1975) in the listing mentioned
below. For another synthetic procedure for making the
menthanecarboxylic acid, see D. Cunningham, et. al., J. Chem. Soc.,
Perkin Trans. I, 2002, 2692-2698.
[0136] For additional preparative details for making cool-genic
compounds mentioned in J. Soc. Cosmet. Chem., 1978, 29, 185-200,
see on page 199, reference 1, listing of 17 U.K. Patents. Other
conditions suitable for formation of the compounds of the
disclosure from a variety of intermediates as illustrated herein
are well known to the art. For example, see Feiser and Feiser,
"Reagents for Organic Synthesis," Vol. 1, 1967; March, J. "Advanced
Organic Chemistry," 4.sup.th ed., John Wiley & Sons, 1992;
House, H. O., "Modern Synthetic Reactions", 2.sup.nd ed., W. A.
Benjamin, New York, 1972; and Larock, R. C., "Comprehensive Organic
Transformations," 2.sup.nd ed., Wiley-VCH Publishers, New York,
1999.
[0137] The starting materials employed in the synthetic methods
described herein are commercially available, have been reported in
the scientific literature, or can be prepared from readily
available starting materials using procedures known in the field.
It may be desirable to optionally use a protecting group during all
or portions of the above described or alternative synthetic
procedures. Such protecting groups and methods for their
introduction and removal are well known in the art. See Greene, T.
W.; Wutz, P. G. M. "Protecting Groups In Organic Synthesis"
2.sup.nd ed., New York, John Wiley & Sons, Inc., 1991.
[0138] In cases where compounds are sufficiently basic or acidic to
form stable nontoxic acid or base salts, administration of the
compound of the disclosure as a salt may be appropriate. Examples
of pharmaceutically acceptable salts are organic acid addition
salts formed with acids, which form a physiological acceptable
anion, for example, tosylate, methanesulfonate, acetate, citrate,
malonate, tartarate, succinate, benzoate, ascorbate,
.alpha.-ketoglutarate, and .alpha.-glycerophosphate. Suitable
inorganic salts may also be formed, including hydrochloride,
hydrobromide, sulfate, nitrate, bicarbonate, and carbonate
salts.
[0139] Pharmaceutically acceptable salts may be obtained using
standard procedures well known in the art, for example, by reacting
a sufficiently basic compound such as an amine with a suitable acid
affording a physiologically acceptable anion. Alkali metals, for
example, sodium, potassium or lithium, or alkaline earth metal
salts, for example, calcium, of carboxylic acids can also be
made.
[0140] Compounds of the present disclosure can conveniently be
administered in a pharmaceutical composition containing the
compound in combination with a suitable excipient, the composition
being useful in treating tumors. Pharmaceutical compositions
containing a compound appropriate for anti-tumor use are prepared
by methods and contain excipients well known in the art. A
generally recognized compendium of such methods and ingredients is
Remington's Pharmaceutical Sciences by E. W. Martin (Mark Publ.
Co., 15.sup.th ed., 1975). The compounds and compositions of the
present disclosure can be administered parenterally, for example,
by intravenous, intraperitoneal or intramuscular injection, orally,
or rectally, depending on, for example, the disposition or
dissemination of the tumor cells.
[0141] In embodiments, the antibodies included within the scope of
the disclosure include hybrid and recombinant antibodies (e.g.,
"humanized" and "human" antibodies) regardless of species of origin
or immunoglobulin class or subclass designation, as well as
antibody fragments (for example, Fab, F(ab').sub.2, and F.sub.v),
See U.S. Pat. No. 4,816,567; Mage and Lamoyi, in Monoclonal
Antibody Production Techniques and Applications, 79-97, Marcel
Dekker, Inc., New York, (1987).
[0142] Thus, the modifier "monoclonal" indicates the character of
the antibody as being obtained from such a substantially
homogeneous population of antibodies, and is not to be construed as
requiring production of the antibody by any particular method. For
example, the monoclonal antibodies of the disclosure may be made
using the hybridoma method first described by Kohler &
Milstein, Nature, 256:495 (1975), or may be made by recombinant DNA
methods. See U.S. Pat. No. 4,816,567. Other known methods of
antibody production are described, for example, in Goding,
Monoclonal Antibodies: Principles and Practice, 59-103, Academic
Press (1986); Kozbor, J. Immunol., 133:3001 (1984). Brodeur, et
al., Monoclonal Antibody Production Techniques and Applications,
51-63, Marcel Dekker, Inc., New York (1987).
[0143] Various methods have been employed to produce monoclonal
antibodies (MAbs). Hybridoma technology, which refers to a cloned
cell line that produces a single type of antibody, uses the cells
of various species, including mice (murine), hamsters, rats, and
humans. Another method to prepare MAbs uses genetic engineering
including recombinant DNA techniques. Monoclonal antibodies made
from these techniques include, among others, chimeric antibodies
and humanized antibodies. A chimeric antibody combines DNA encoding
regions from more than one type of species. For example, a chimeric
antibody may derive the variable region from a mouse and the
constant region from a human. A humanized antibody comes
predominantly from a human, even though it contains nonhuman
portions. Like a chimeric antibody, a humanized antibody may
contain a completely human constant region. But unlike a chimeric
antibody, the variable region may be partially derived from a
human. The nonhuman, synthetic portions of a humanized antibody
often come from CDRs in murine antibodies. In any event, these
regions are crucial to allow the antibody to recognize and bind to
a specific antigen.
[0144] As noted, murine antibodies play an important role in these
technologies. While useful for diagnostics and short-term
therapies, murine antibodies cannot be administered to people
long-term without increasing the risk of a deleterious immunogenic
response. This response, called Human Anti-Mouse Antibody (HAMA),
occurs when a human immune system recognizes the murine antibody as
foreign and attacks it. A HAMA response can cause toxic shock or
even death.
[0145] Chimeric and humanized antibodies reduce the likelihood of a
HAMA response by minimizing the nonhuman portions of administered
antibodies. Furthermore, chimeric and humanized antibodies have the
additional benefit of activating secondary human immune responses,
such as antibody dependent cellular cytotoxicity.
[0146] "Antibody fragments" comprise a portion of an intact
antibody, preferably comprising the antigen-binding or variable
region thereof. Examples of antibody fragments include Fab, Fab',
F(ab')2, and Fv fragments; diabodies; linear antibodies;
single-chain antibody molecules; and multi-specific antibodies
formed from antibody fragment(s).
[0147] An "intact" antibody is one which comprises an
antigen-binding variable region as well as a light chain constant
domain (CL) and heavy chain constant domains, CH.sub.1, CH.sub.2
and CH.sub.3. The constant domains may be native sequence constant
domains (e.g., human native sequence constant domains) or amino
acid sequence variant thereof.
[0148] The intact antibody may have one or more "effector
functions" which refer to those biological activities attributable
to the Fc region (a native sequence Fc region or amino acid
sequence variant Fc region) of an antibody. Examples of antibody
effector functions include Clq binding; complement dependent
cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated
cytotoxicity (ADCC); phagocytosis; down regulation of cell surface
receptors (e.g., B cell receptor; BCR), etc. Depending on the amino
acid sequence of the constant domain of their heavy chains, intact
antibodies can be assigned to different "classes." There are five
major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM,
and several of these may be further divided into "subclasses"
(isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The
heavy-chain constant domains that correspond to the different
classes of antibodies are called .alpha., .delta., .epsilon.,
.gamma., and .mu., respectively. The subunit structures and
three-dimensional configurations of different classes of
immunoglobulins are well known.
[0149] When used in vivo for combination therapy, antibodies can be
administered to the patient in therapeutically effective amounts
(i.e. amounts that eliminate or reduce the patient's tumor burden).
The combination of a compound of the disclosure and antibodies can
be administered at the same time or sequentially. They will
normally be administered parenterally, when possible, at the target
cell site, or intravenously. The dose and dosage regimen will
depend upon, for example, the nature of the cancer (primary or
metastatic), its population, the site to which the antibodies are
to be directed, the characteristics of the particular immunotoxin
(when used), for example, its therapeutic index, the patient, and
the patient's history. The amount of antibody administered will
typically be in the range of about 0.1 to about 10 mg/kg of patient
weight. The amount of a compound of the disclosure administered in
combination with an antibody can typically be, for example, in the
range of about 5 to 1,000 mg, or about 0.1 to 300 mg/kg of patient
body weight, and can depend on many of the abovementioned factors
and considerations.
[0150] The present disclosure also contemplates using combinations
of a compound of the disclosure with an anti-TRP-P8 antibody in
diagnostic applications. For diagnostic applications, the
antibodies of the disclosure typically will be labeled with a
detectable moiety. The detectable moiety can be any one capable of
producing, either directly or indirectly, a detectable signal. For
example, the detectable moiety may be a radioisotope, such as
.sup.3H, .sup.14C, .sup.32P, .sup.35S, or .sup.125I, a fluorescent
or chemiluminescent compound, such as fluorescein isothiocyanate,
rhodamine, or luciferin; or an enzyme, such as alkaline
phosphatase, beta-galactosidase or horseradish peroxidase.
[0151] Any method known in the art for separately conjugating the
antibody to the detectable moiety may be employed, including those
methods described by Hunter, et al., Nature, 144:945 (1962); David,
et al., Biochemistry, 13:1014 (1974); Pain, et al., J. Immunol.
Meth., 40:219 (1981); and Nygren, J. Histochem. and Cytochem,
30:407 (1982).
[0152] For therapeutic applications, antibodies may be administered
to a mammal, preferably a human, in a pharmaceutically acceptable
dosage form, including those that may be administered to a human
intravenously as a bolus or by continuous infusion over a period of
time, by intramuscular, subcutaneous, intra-articular, or
inhalation routes. An antibody is also suitably administered by
intra-tumoral, peritumoral, intra-lesional, or peri-lesional
routes, to exert local as well as systemic therapeutic effects.
[0153] Such dosage forms encompass known pharmaceutically
acceptable carriers or vehicles that are inherently nontoxic and
non-therapeutic. An antibody will typically be formulated in such
vehicles at a concentration of about 0.1 mg/ml to 100 mg/ml.
[0154] For the prevention or treatment of disease, the appropriate
dosage of a combination of a compound of the disclosure will depend
on the type of disease to be treated, as defined above, the
severity and course of the disease, whether the compound is
administered for preventive or therapeutic purposes, previous
therapy, the patient's clinical history and response to the
compound, and the discretion of the attending physician. The
compound is suitably administered to the patient at one time or
over a series of treatments.
[0155] Depending on the type and severity of the disease, about
0.015 to 15 mg/kg of the compound is an initial candidate dosage
for administration to the patient, whether, for example, by one or
more separate administrations, or by continuous infusion. For
repeated administrations over several days or longer, depending on
the condition, the treatment is repeated until a desired
suppression of disease symptoms occurs. However, other dosage
regimens may be useful.
[0156] For oral therapeutic administration, the active compound of
the disclosure may be combined with one or more excipients and used
in the form of ingestible tablets, buccal tablets, troches,
capsules, elixirs, suspensions, syrups, wafers, and the like. Such
compositions and preparations typically contain at least about 0.1%
of active compound. The percentage of the compositions and
preparations may, of course, be varied and may conveniently be
between about 2 to about 60% of the weight of a given unit dosage
form. The amount of active compound in such therapeutically useful
compositions is such that an effective dosage level will be
obtained.
[0157] The tablets, troches, pills, capsules, and the like may also
contain the following: binders such as gum tragacanth, acacia, corn
starch or gelatin; excipients such as dicalcium phosphate; a
disintegrating agent such as corn starch, potato starch, alginic
acid and the like; a lubricant such as magnesium stearate; and a
sweetening agent such as sucrose, fructose, lactose or aspartame or
a flavoring agent such as peppermint, oil of wintergreen, or cherry
flavoring may be added. When the unit dosage form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier, such as a vegetable oil or a polyethylene glycol. Various
other materials may be present as coatings or to otherwise modify
the physical form of the solid unit dosage form. For instance,
tablets, pills, or capsules may be coated with gelatin, wax,
shellac, or sugar, and the like. A syrup or elixir may contain the
active compound, sucrose or fructose as a sweetening agent, methyl
and propylparabens as preservatives, a dye and flavoring such as
cherry or orange flavor. Of course, any material used in preparing
any unit dosage form should be pharmaceutically acceptable and
substantially non-toxic in the amounts employed. In addition, the
active compound may be incorporated into sustained-release
preparations and devices.
[0158] The compounds or compositions of the disclosure can also be
administered intravenously or intraperitoneally by infusion or
injection. Solutions of the active compound or its salts can be
prepared in water, optionally mixed with a nontoxic surfactant.
Dispersions can also be prepared in glycerol, liquid polyethylene
glycols, triacetin, and mixtures thereof and in oils. Under
ordinary conditions of storage and use, these preparations contain
a preservative to prevent the growth of microorganisms.
[0159] Pharmaceutical dosage forms suitable for injection or
infusion can include sterile aqueous solutions or dispersions or
sterile powders comprising the active ingredient, which are adapted
for the extemporaneous preparation of sterile injectable or
infusible solutions or dispersions, optionally encapsulated in
liposomes or implantable seeds or pellets. In all cases, the
ultimate dosage form should be sterile, fluid and stable under the
conditions of manufacture and storage. The liquid carrier or
vehicle can be a solvent or liquid dispersion medium comprising,
for example, water, ethanol, a polyol (for example, glycerol,
propylene glycol, liquid polyethylene glycols, and the like),
vegetable oils, nontoxic glyceryl esters, and suitable mixtures
thereof. The proper fluidity can be maintained, for example, by the
formation of liposomes, by the maintenance of the required particle
size in the case of dispersions or by the use of surfactants. The
prevention of the action of microorganisms can be brought about by
various antibacterial and antifungal agents, for example, parabens,
chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In
many cases, it will be preferable to include isotonic agents, for
example, sugars, buffers or sodium chloride. Prolonged absorption
of the injectable compositions can be brought about by the use in
the compositions of agents delaying absorption, for example,
aluminum monostearate and gelatin.
[0160] Sterile injectable solutions can be prepared by
incorporating the active compound in the required amount in the
appropriate solvent with various of the other ingredients
enumerated above, as required, followed by filter sterilization. In
the case of sterile powders for the preparation of sterile
injectable solutions, the preferred methods of preparation are
vacuum-drying and freeze-drying techniques, which yield a powder of
the active ingredient plus any additional desired ingredient
present in the previously sterile-filtered solutions.
[0161] Useful solid carriers include finely divided solids such as
talc, clay, microcrystalline cellulose, silica, alumina and the
like. Useful liquid carriers include water, alcohols or glycols or
water-alcohol/glycol blends, in which the present compounds can be
dissolved or dispersed at effective levels, optionally with the aid
of non-toxic surfactants. Adjuvants such as fragrances and
additional antimicrobial agents can be added to optimize the
properties for a given use. Useful dosages of the compounds of the
disclosure can be determined by comparing their in vitro activity,
and in vivo activity in animal models. Methods for the
extrapolation of effective dosages in mice, and other animals, to
humans are known to the art; for example, see U.S. Pat. No.
4,938,949.
[0162] The compound is conveniently administered in unit dosage
form; for example, containing 5 to 1,000 mg, conveniently 10 to 750
mg, most conveniently, 50 to 500 mg of active ingredient per unit
dosage form. The desired dose may conveniently be presented in a
single dose or as divided doses administered at appropriate
intervals, for example, as two, three, four or more sub-doses per
day. The sub-dose itself may be further divided, e.g., into a
number of discrete loosely spaced administrations; such as multiple
inhalations from an insufflator.
[0163] For internal administration, the compositions can be
administered orally or parenterally at dose levels, calculated as
the free base, of about 0.1 to 300 mg/kg, preferably 1.0 to 30
mg/kg of mammal body weight, and can be used in man in a unit
dosage form, administered one to four times daily in the amount of
1 to 1,000 mg per unit dose.
[0164] For parenteral administration or for administration as
drops, as for eye treatments, the compounds are presented in
aqueous solution in a concentration of from about 0.1 to about 10%,
more preferably about 0.1 to about 7%. The solution may contain
other ingredients, such as emulsifiers, antioxidants or
buffers.
[0165] Generally, the concentration of the compound(s) of formula
(I-XIII) in a liquid composition, such as an IV (intravenous), will
be from about 0.1 to about 25, preferably from about 0.5 to about
10, weight percent. The concentration in a semi-solid or solid
composition such as a gel or a powder will be about 0.1 to about 5
weight percent, preferably about 0.5 to about 2.5 weight
percent.
[0166] The exact regimen for administration of the compounds and
compositions disclosed herein will necessarily be dependent upon
the needs of the individual subject being treated, the type of
treatment and, of course, the judgment of the attending
practitioner.
[0167] The binding activity and binding selectivity of the
compounds of the present disclosure are excellent predictors of the
anti-tumor activity of compounds of the disclosure. The binding
activity and binding selectivity can be determined using
pharmacological models well known to the art, or using the assays
described below. Exemplary results of biological testing are
summarized in Table 1 below.
Evaluation of Biological Activity
[0168] General methods and materials disclosed in R. Skyryma, et
al., J. Physiology, 2000, 527.1, 71-83, for assessing and measuring
calcium ion flux or cell uptake, and induction of apoptosis were
adapted for use in the present disclosure and as illustrated
herein. Other test methods and procedures such as described below,
including cell line culturing, transfection, and in vivo and in
vitro tumor growth inhibition, are readily apparent to one of
ordinary skill in the art upon comprehending the disclosure.
Assays for Activity
[0169] For cancer, a variety of well-known animal models can be
used to further understand the role of the genes in the development
and pathogenesis of tumors, and to test the efficacy of candidate
therapeutic agents, including combinations of compounds of the
disclosure and anti-angiogenic antibodies. The in vivo nature of
such models makes them particularly predictive of responses in
human patients. Animal models of tumors and cancers (e.g., breast
cancer, colon cancer, prostate cancer, lung cancer, etc.) include
both non-recombinant and recombinant (transgenic) animals.
Non-recombinant animal models include, for example, rodent, e.g.,
murine models. Such models can be generated by introducing tumor
cells into syngeneic mice using standard techniques, e.g.,
subcutaneous injection, tail vein injection, spleen implantation,
intraperitoneal implantation, implantation under the renal capsule,
or orthopin implantation, e.g., colon cancer cells implanted in
colonic tissue. See for example, PCT publication No. WO 97/33551,
published Sep. 18, 1997.
[0170] Probably the most often used animal species in oncological
studies are immunodeficient mice and, in particular, nude mice. The
observation that the nude mouse with thymic hypo/aplasia could
successfully act as a host for human tumor xenografts has lead to
its widespread use for this purpose. The autosomal recessive nu
gene has been introduced into a very large number of distinct
congenic strains of nude mouse, including, for example, ASW, A/He,
AKR, BALB/c, B10.LP, C17, C3H, C57BL, C57, CBA, DBA, DDD, 1/st, NC,
NFR, NFS, NFS/N, NZB, NZC, NZW, P, RIII, and SJL. In addition, a
wide variety of other animals with inherited immunological defects
other than the nude mouse have been bred and used as recipients of
tumor xenografts. For further details see for example, The Nude
Mouse in Oncology Research, E. Boven and B. Winograd, eds. (CRC
Press, Inc., 1991).
[0171] The cells introduced into such animals can be derived from
known tumor/cancer cell lines, such as any of the above-listed
tumor cell lines, and, for example, the B104-1-1 cell line (stable
NIH-3T3 cell line transfected with the neu protooncogene);
ras-transfected NIH-3T3 cells; Caco-2 (ATCC HTB-37); or a
moderately well-differentiated grade II human colon adenocarcinoma
cell line, HT-29 (ATCC HTB-38); or from tumors and cancers. Samples
of tumor or cancer cells can be obtained from patients undergoing
surgery, using standard conditions involving freezing and storing
in liquid nitrogen. Karmali et al., Br. J. Cancer, 48: 689-696
(1983). Tumor cells can be introduced into animals such as nude
mice by a variety of procedures. The subcutaneous (s.c.) space in
mice is very suitable for tumor implantation. Tumors can be
transplanted s.c. as solid blocks, as needle biopsies by use of a
trochar, or as cell suspensions. For solid-block or trochar
implantation, tumor tissue fragments of suitable size are
introduced into the s.c. space. Cell suspensions are freshly
prepared from primary tumors or stable tumor cell lines, and
injected subcutaneously. Tumor cells can also be injected as
subdermal implants. In this location, the inoculum is deposited
between the lower part of the dermal connective tissue and the s.c.
tissue.
[0172] Animal models of breast cancer can be generated, for
example, by implanting rat neuroblastoma cells (from which the neu
oncogene was initially isolated), or neu-transformed NIH-3T3 cells
into nude mice, essentially as described by Drebin, et al., Proc.
Nat. Acad. Sci. USA, 83: 9129-9133 (1986).
[0173] Similarly, animal models of colon cancer can be generated by
passaging colon cancer cells in animals, e.g., nude mice, leading
to the appearance of tumors in these animals. An orthotopic
transplant model of human colon cancer in nude mice has been
described, for example, by Wang, et al., Cancer Research, 54:
4726-4728 (1994) and Too, et al., Cancer Research, 55: 681-684
(1995). This model is based on the so-called "METAMOUSE".TM. sold
by AntiCancer, Inc. (San Diego, Calif.).
[0174] Tumors that arise in animals can be removed and cultured in
vitro. Cells from the in vitro cultures can then be passaged to
animals. Such tumors can serve as targets for further testing or
drug screening. Alternatively, the tumors resulting from the
passage can be isolated and RNA from pre-passage cells and cells
isolated after one or more rounds of passage analyzed for
differential expression of genes of interest. Such passaging
techniques can be performed with any known tumor or cancer cell
lines. The following examples serve to more fully describe the
manner of using the above-described disclosure, as well as to set
forth the best modes contemplated for carrying out various aspects
of the disclosure. It is understood that these examples in no way
serve to limit the true scope of this disclosure, but rather are
presented for illustrative purposes.
Example 1
[0175] Preparation of Compound IIa-4-1 from (-)Menthyl chloride.
1.39 grams of Mg metal (57 mmol) was placed in a 100 mL flask and 4
mL of dry THF was added to cover the Mg metal. A crystal of iodine
was added to the metal-THF mixture and stirred for several minutes
followed by the addition of about 1/3 portion of 10 grams (57 mmol)
of (-) menthyl chloride (Aldrich). The mixture was heated to induce
Grignard formation and the remaining menthyl chloride (2/3 portion)
was added in 50 mL dry THF and stirred until the reaction was
complete. The Grignard solution was then canulated under nitrogen
into a vessel containing excess dry ice. This dry ice quenched
mixture was swirled and poured into 300 mL ice containing 2 mL
concentrated HCl. Diethyl ether was added and the mixture
separated. The separated organic layers were combined and washed
with water then extracted with an aqueous NaOH solution. The
aqueous solution (and oil) was acidified with HCl until a solid
formed, diethyl ether was added, and the acid was extracted into
the organic phase, washed with brine, dried over sodium sulfate,
and concentrated to give 4.18 grams (40%) as white needles.
[0176] The acid (0.57 grams, 3.1 mmol) was dissolved in 1.5 mL
dimethylacetamide (DMA). The PyBOP (2.1 grams, 4.0 mmol),
p-anisidine (0.58 grams, 4.7 mmol, Aldrich) and DiPEA (2.7 mL, 15.5
mmol) were added. Another 0.25 grams p-anisidine was added after 2
hours and again after 5 hours. The reaction was diluted with ethyl
acetate (EtOAc) after 7 hours, washed twice with 1 N HCl, twice
with aqueous sodium bicarbonate, and then with brine. The separated
organic layers were combined and dried over sodium sulfate and
concentrated to a brown solid. This solid was put through a plug of
silica gel with 30% EtOAc/hexanes to remove the color. The product
began to crystallize upon concentration, so the crystallization was
allowed to occur, the solvent was removed by pipette, and the white
needles were washed with cold EtOAc and dried under vacuum to give
0.365 grams (1.26 mmol, 40% yield) of a first crop.
Recrystallization of the mother liquor yielded an additional 0.346
grams (1.2 mmol, 39% yield) as white needles.
[0177] LCMS showed the product to have a molecular weight of 289,
corresponding to the desired molecular weight, and the NMR spectra
showed it to have the desired structure.
Example 2
[0178] Demonstration of Ion Channel Activity in Response to
Selected Cool-Genic Compounds--Calcium Ion Uptake Evaluation.
Calcium ion uptake experiments were conducted as follows. Calcium
uptake was measured for cells expressing Trp-p8 tumor antigen and
for cells not expressing Trp-p8 tumor antigen after each cell line
was contacted with cool-genic compounds of the present disclosure.
The results showed that cells expressing Trp-p8 tumor antigen had
progressively increased calcium uptake as the concentration of
cool-genic compound was increased incrementally over about five
concentration decades from about 0.0001 to about 10 microM.
Compound (IIa-4-1) had unexpectedly particularly high calcium
uptake at all cool-genic compound concentrations. Cells not
expressing tumor antigen had essentially no observable calcium
uptake over all of the cool-genic compound concentrations. Dimethyl
sulfoxide (DMSO), a non-coolgenic compound, was used as a control
compound in both expressing and non-expressing cell lines and which
DMSO exposed cells showed no appreciable calcium uptake at any
concentration. Cell lines included non-expressing lines: 293, PC3,
and PC3/neo; and tumor antigen expressing lines: 293, PC3, and
PC3/neo.
Example 3
[0179] Human Cell Killing in vitro by Compounds that Activate
Trp-p8. Referring to the Figures, in FIG. 1 the effectiveness of
selected cool-genic compounds in human cell killing was evaluated.
The order and potency of cool-genic compounds were compared in two
related human cell lines, one was 293 cells expressing Trp-p8 and a
second was matched 293 cells lacking (not expressing) Trp-p8 used
as a control. There were about 50,000 cells per well at plating.
The cool-genic compounds were added at plating. The cells were
treated by exposing the cells to the individual compounds at
varying concentrations over about 0.1 to about 1,000 microM for 72
hours. The x-axis shows concentration increments and the y-axis
shows the number (in arbitrary units) of viable cells remaining at
the end of 72 hours. FIGS. 1A and 1C show the response of cells
expressing Trp-p8 and treated with the indicated numbered or named
coolgenic compounds. FIGS. 1B and 1D show the response of cells
without (not expressing) Trp-p8 and also treated with the indicated
coolgenic compounds. The results in FIGS. 1A and 1C show that the
cells with Trp-p8 had a range of kill responses or potencies with
respect to the coolgenic compounds. Compound IIa-4-1 had the best
kill (IC.sub.50<1 microM) for cells expressing Trp-p8. The
comparative results in FIGS. 1B and 1D show that the cells without
Trp-p8 had a lower or essentially no kill response to the same
coolgenic compounds, particularly at the lower concentrations of
the coolgenic compounds, e.g., below 100 microMolar.
Example 4
[0180] Procedure for inhibition of tumor growth in vivo by
compounds that activate Trp-p8. Methods for preparing transfected
cancerous cells, such as those expressing Trp-p8, is readily
apparent to one of ordinary skill in the art, see for example J. M.
Schallhorn, et al., Nucleic Acids Res., 1996, Feb. 15;
24(4):596-601. Referring to the FIG. 2, there is illustrated the
growth of PC3 clone cells transfected to express tumor antigen,
(PC3/Trp-p8.c8 and .c9), and PC3 clone cells not expressing tumor
antigen (PC3-Neo) in athymic nude mice (5 million cells/mouse). The
non-Trp-p8 expressing tumor forming human prostate cancer cell
line, PC3-Neo 200 (- -), shows expected exponential tumor cell
growth rates. In contrast, the Trp-p8 expressing (i.e.
"over-expressing") tumor forming human prostate cancer cell lines,
PC3/Trp-p8: .c8 (clone 8) 220 (-.box-solid.-); and .c9 (clone 9)
230 (-.tangle-solidup.-), show substantially slower growth and
static growth rates, respectively.
[0181] Athymic mice (8 per group) are inoculated in the flank with
about 5.times.10.sup.6 of PC3 clone cells stablely expressing
Trp-p8 (c1.8) or a vector control cell line (PC3-Neo). Tumors are
established to a size of approximately 200 mg each at which time
coolgenic compounds are administered once or twice per day I.V. or
P.O. at doses ranging from about 1-30 mg/kg body weight. Tumor
volumes are measured by caliper every third day and average volumes
are calculated.
[0182] Contacting, in vivo, the Trp-p8 expressing cancerous cells,
such as those illustrated above, with certain coolgenic compounds
of moderate to high potency of the present disclosure, and as
illustrated herein, is expected to result in high cell kill for
Trp-p8 expressing cancerous cells and no cell kill or low cell kill
for healthy or non-Trp-p8 expressing cells. This expectation is
consistent with the abovementioned in vitro human cell killing
results presented in EXAMPLE 3.
[0183] Table 1 summarizes exemplary ion channel activity and Cell
Kill response to selected cool-genic compounds, mentioned above or
illustrated below, and provides a comparative or relative activity
ranking for those compounds.
TABLE-US-00001 TABLE 1 Ion Channel Activity and Cell Killing in
Response to Selected Cool-Genic Compounds. Activity Compound
293/Trp-p8 PC3/Trp-p8 Ranking ID# Peak (FLIPR).sup.1 Cell Killing
Cell Killing 1 IIa4-1 31,000 Yes Yes 2 IV-1 31,000 Yes 3 XIII-1
29,000 Yes Yes 4 IV-2 29,000 5 IIb-2 25,100 Yes Yes 6 IIb-3 25,100
Yes Yes 7 IIb-4 25,100 Yes 8 IIb-1 25,000 9 XIII-2 25,000 10 IIb-5
23,300 11 IV-3 22,000 12 Icilin 20,000 13 V-4 17,500 14 V-2 17,000
15 V-1 9,600 16 III-1 5,000 17 DMSO 1,400 control .sup.1Peak
(FLIPR) is a measure of maximum calcium ion flux at 10 microM.
FLIPR refers to Fluorometric Imaging Plate Reader; commercially
available from, for example, Molecular Devices Corp.
##STR00037## ##STR00038## ##STR00039##
Example 5
[0184] Preparation of Other Selected Compounds of Formula IIa.
Example I was generally repeated for each of the following
preparative compound examples with the exception that the amine
(p-anisidine) co-reactant of Example I was substituted with the
corresponding amine to produce compound having the structure
indicated as the major product upon purification. The major product
for each of the following examples was characterized by, for
example, mass spectra to have a parent peak at about the molecular
weight indicated.
##STR00040## ##STR00041## ##STR00042## ##STR00043##
[0185] Table 2 summarizes IC.sub.50 results for the indicated
menthane carboxamide compounds on 293/Trp-p8.clone 18 and
293/Trp-p8.clone 10.
TABLE-US-00002 TABLE 2 IC.sub.50 Data for Menthane Carboxamides on
293/Trp-p8.clone 18 and 293/Trp-p8.clone 10. Compound IIa2 where
R.sub.1 = H and R.sub.2 = Compound
--PhR.sub.3R.sub.4R.sub.5R.sub.6R.sub.7 is ID# 293/Trp-p8.c18
293/Trp-p8.c10 4-MeO--Ph-- IIa4-1 0.58/0.45 0.42 Ph-- IIa4-2 3.38
3.21 4-HO--Ph-- IIa4-3 1.05 1.23 3-MeO--Ph-- IIa10-1 3.69 3.80
4-iPr--Ph-- IIa4-4 0.98 0.76 2-MeO--Ph-- IIa8-1 17.57 16.74
4-Cl--Ph-- IIa4-5 17.57 1.99 4-F--Ph-- IIa4-6 3.78 3.58 3-F,
4-MeO--Ph-- IIa6-1 0.60 0.79 4-Me--Ph-- IIa4-7 1.14 1.21 4-Et--Ph--
IIa4-8 0.68 0.73 2-Me, 4-MeO--Ph-- IIa12-1 0.93 1.14 3-F--Ph--
IIa10-2 2.91 6.31 4-morpholino-Ph-- IIa4-9 1.45 1.28 3-Cl,
4-Me--Ph-- IIa6-2 2.57 3.10 4-secBu--Ph-- IIa4-10 1.08 3.59
3,4-propyleneyl-Ph-- IIa6-3 1.71 1.35 (i.e., indanyl) 4-tBuPh--
IIa4-11 1.41 1.23 4-nPrPh-- IIa4-12 4.93 5.65 3-Me, 4-iPrPh--
IIa6-4 4.37 5.05 Icilin (reference) -- -- 79.24
Example 6
[0186] Table 3 summarizes IC.sub.50 duplicate results for the
indicated menthane carboxamide compounds on 293/Trp-p8.clone
21.
TABLE-US-00003 TABLE 3 IC.sub.50 Data for Menthane Carboxamides on
PC3/Trp-p8.clone 21 (in duplicate). Compound ID# IC.sub.50 Compound
IIa4 where R5- is 4-MeO--Ph-- IIa4-1 1.642 4-HO--Ph-- IIa4-3 4.2696
4-iPr--Ph-- IIa4-4 3.2082 4-secBu--Ph-- IIa4-10 8.4937 4-nPr--Ph--
IIa4-12 12.857 Compound IIa4 where R5 is 4-MeO--Ph-- IIa4-1 1.5938
4-HO--Ph-- IIa4-3 3.7606 4-iPr--Ph-- IIa4-4 3.1039 4-secBu--Ph--
IIa4-10 48.613 4-nPr--Ph-- IIa4-12 16.473
Example 7
[0187] Carrier articles, such as commercially available or custom
manufactured implantable seeds or pellets, can be formulated and
impregnated with one or more compound of the present disclosure,
alone or in combination with another chemotherapeutic agent or
other medicaments or excipients. A preferred formulation can have,
for example, selectable timed-release characteristics for the
coolgenic compound or other ingredients, for example, for use in
prostate cancer treatment. For an example of non-radioactive
sustained release implants in therapeutic cancer treatment, such as
prostate cancer, see U.S. Pat. No. 5,633,274, which disclosure is
incorporated herein by reference in its entirety.
Example 8
[0188] The following illustrate representative pharmaceutical
dosage forms, containing a compound of the disclosure (`Compound
x`), such as a compound of formula I or II, for therapeutic or
prophylactic use in humans.
TABLE-US-00004 (i) Tablet 1 mg/tablet `Compound x` 100.0 Lactose
77.5 Povidone 15.0 Croscarmellose sodium 12.0 Microcrystalline
cellulose 92.5 Magnesium stearate 3.0 300.0
TABLE-US-00005 (ii) Tablet 2 mg/tablet `Compound x` 20.0
Microcrystalline cellulose 410.0 Starch 50.0 Sodium starch
glycolate 15.0 Magnesium stearate 5.0 500.0
TABLE-US-00006 (iii) Capsule mg/capsule `Compound x` 10.0 Colloidal
silicon dioxide 1.5 Lactose 465.5 Pregelatinized starch 120.0
Magnesium stearate 3.0 600.0
TABLE-US-00007 (iv) Injection 1 (1 mg/ml) mg/ml `Compound x` 1.0
Dibasic sodium phosphate 12.0 Monobasic sodium phosphate 0.7 Sodium
chloride 4.5 1.0N Sodium hydroxide solution q.s. (pH adjustment to
7.0-7.5) Water for injection q.s. ad 1 mL
TABLE-US-00008 (v) Injection 2 (10 mg/ml) mg/ml `Compound x` 10.0
Monobasic sodium phosphate 0.3 Dibasic sodium phosphate 1.1
Polyethylene glycol 400 200.0 01N Sodium hydroxide solution q.s.
(pH adjustment to 7.0-7.5) Water for injection q.s. ad 1 mL
TABLE-US-00009 (vi) Aerosol mg/can `Compound x` 20.0 Oleic acid
10.0 Trichloromonofluoromethane 5,000.0 Dichlorodifluoromethane
10,000.0 Dichlorotetrafluoroethane 5,000.0
[0189] The above formulations may be obtained by conventional
procedures well known in the pharmaceutical art.
Example 9
[0190] Combination Therapy-Coadministration. The following
illustrates representative pharmaceutical dosage forms, containing
a compound of the disclosure in direct combination (admixture) with
an antibody (collectively `Composition y`), for therapeutic or
prophylactic use in humans. Thus, for example, a compound of the
disclosure, such as a compound of formula I or II, is combined with
an antibody, such as an anti-VEGF antibody. The resulting
combination, `Composition y`, is substituted in place of `Compound
x` in one or more of the above-mentioned injection formulations of
EXAMPLE 8. The above formulations may be obtained by conventional
procedures well known in the pharmaceutical arts.
Example 10
[0191] Combination Therapy--Serial Administration. The following
illustrates representative pharmaceutical dosage forms, containing
a compound of the disclosure (`Compound x`) in serial combination
with an antibody (`antibody z`), for therapeutic or prophylactic
use in humans. Thus, a compound of the disclosure, such as a
compound of formula I or II, is serially administered with an
antibody, such as an anti-VEGF antibody. For example, the serially
administered combination can include first administration of a
compound of the disclosure (`Compound x`) is any of the above
mentioned formulations of EXAMPLE 7 or 8 followed by a second
injection administration of an antibody (`antibody z`).
Alternatively, `antibody z` is administered first followed by the
administration of `Compound x`. The above formulations may be
obtained by conventional procedures well known in the
pharmaceutical arts.
Example 11
[0192] Methods, examples, and additional literature references for
the preparation of antibodies, and characterization of antibodies,
including antigen specificity, epitope mapping, isotyping, binding
affinity, are disclosed in the aforementioned U.S. Pat. No.
6,582,959.
[0193] All publications, patents, and patent documents are
incorporated by reference herein in their entirety, as though
individually incorporated by reference. The disclosure has been
described with reference to various specific and preferred
embodiments and techniques. However, it should be understood that
many variations and modifications can be made while remaining
within the spirit and scope of the disclosure.
* * * * *