U.S. patent application number 14/217929 was filed with the patent office on 2014-07-17 for method for administration.
The applicant listed for this patent is LIN-CHI CHEN, BRIAN HIGGINS, GWEN NICHOLS. Invention is credited to LIN-CHI CHEN, BRIAN HIGGINS, GWEN NICHOLS.
Application Number | 20140200255 14/217929 |
Document ID | / |
Family ID | 51165610 |
Filed Date | 2014-07-17 |
United States Patent
Application |
20140200255 |
Kind Code |
A1 |
HIGGINS; BRIAN ; et
al. |
July 17, 2014 |
METHOD FOR ADMINISTRATION
Abstract
There is provided a new dosage regimen for Compound A which
maximizes anti-tumor activity while maintaining acceptable toxicity
levels.
Inventors: |
HIGGINS; BRIAN; (FRESH
MEADOWS, NY) ; NICHOLS; GWEN; (NEW YORK, NY) ;
CHEN; LIN-CHI; (NEW YORK, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
HIGGINS; BRIAN
NICHOLS; GWEN
CHEN; LIN-CHI |
FRESH MEADOWS
NEW YORK
NEW YORK |
NY
NY
NY |
US
US
US |
|
|
Family ID: |
51165610 |
Appl. No.: |
14/217929 |
Filed: |
March 18, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13759647 |
Feb 5, 2013 |
|
|
|
14217929 |
|
|
|
|
61612429 |
Mar 19, 2012 |
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Current U.S.
Class: |
514/423 |
Current CPC
Class: |
A61K 9/10 20130101; A61K
31/40 20130101; A61K 9/146 20130101; A61K 9/284 20130101 |
Class at
Publication: |
514/423 |
International
Class: |
A61K 31/40 20060101
A61K031/40 |
Claims
1. A method of treating a patient suffering from cancer, comprising
administering to said patient a pharmaceutical composition
containing as an active ingredient Compound A in an amount from
about 400 mg/day to about 1600 mg/day, daily, for up to about 7
days, followed by a rest period of about 21 days, said
administration starting on the first day of a 28 day treatment
cycle.
2. A method of treating a patient suffering from cancer, comprising
administering to said patient a pharmaceutical composition
containing as an active ingredient Compound A in an amount from
about 400 mg/day to about 1600 mg/day, daily, for up to about 5
days, followed by a rest period of about 23 days, said
administration starting on the first day of a 28 day treatment
cycle.
3. The method of claim 2 wherein Compound A is administered in an
amount from about 400 mg/day to about 1200 mg/day.
4. The method of claim 1 wherein the treatment cycle being repeated
every 28 days for up to about 12 cycles.
5. The method of claim 1 wherein Compound A is administered twice
daily in equal doses.
6. The method of claim 1 wherein the cancer is selected from the
group consisting of colon, breast, prostate, head, neck, lung,
kidney, leukemia, melanoma, soft tissue sarcoma and
osteosarcoma.
7. The method of claim 2, comprising administering to said patient
a pharmaceutical composition containing as an active ingredient
Compound A at about 500 mg/day total, daily for up to about 5 days,
followed by a rest period of about 23 days, said administration
starting on the first day of a 28 day treatment cycle.
8. The method of claim 7 wherein the cancer is selected from the
group consisting of colon, breast, prostate, head, neck, lung,
kidney, melanoma, soft tissue sarcoma and osteosarcoma.
9. A method of treating a patient suffering from leukemia,
comprising administering to said patient a pharmaceutical
composition containing as an active ingredient Compound A at about
1200 mg/day total daily, for up to about 5 days, followed by a rest
period of about 23 days, said administration starting on the first
day of a 28 day treatment cycle.
10. A method of treating a patient suffering from cancer,
comprising administering to said patient a pharmaceutical
composition containing as an active ingredient Compound A in an
amount of about 400-1000 mg total daily dose for up to 3 days
followed by a rest period of up to about 25 days, said
administration starting on the first day of a 28 day treatment
cycle.
11. The method of claim 10 wherein the cancer is selected from the
group consisting of colon, breast, prostate, head, neck, lung,
kidney, leukemia, melanoma, soft tissue sarcoma and osteosarcoma.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation-In-Part of application
Ser. No. 13/759,647, filed Feb. 5, 2013, which claims the benefit
of U.S. Provisional Application No. 61/612,429, filed Mar. 19,
2012. Each of these applications is incorporated by reference
herein in its entirety.
FIELD OF THE INVENTION
[0002] The present invention is related to improved methods of
administration of
4-{[(2R,3S,4R,5S)-4-(4-Chloro-2-fluoro-phenyl)-3-(3-chloro-2-fluoro-pheny-
l)-4-cyano-5-(2,2-dimethyl-propyl)-pyrrolidine-2-carbonyl]-amino}-3-methox-
y-benzoic acid (referred to herein as Compound A) in the treatment
of cancer in humans. In particular, the invention relates to
improved methods of administration of Compound A that provide
desirable antineoplastic effects with a tolerable level of
toxicity. The methods of the invention are characterized by
administering specific doses of Compound A under a specific
protocol which elicits effective and best tolerated biological
activity in human patients. This protocol is expected to be safer
and at least as effective as, possibly more effective than,
administering more frequent doses at lower concentrations or larger
doses at intermittent periods.
BACKGROUND OF THE INVENTION
[0003] P53 is a tumor suppressor that regulates cell cycle,
apoptosis, DNA repair and cell death. In human cancer, the p53
transcription factor is typically inactive leading to uncontrolled
cell growth. Inhibition of p53 activity occurs through binding of
MDM2 to p53. Selective inhibition of the MDM2-p53 interaction
activates the p53 pathway and induces cell cycle arrest and
apoptosis. Compound A is an orally administered pyrrolidine that
inhibits the binding of MDM2 to p53 and is thus useful in the
treatment of cancer. It has the following chemical structure:
##STR00001##
[0004] Compound A recently entered into phase I clinical trials for
the treatment of solid tumors. See ClinicalTrials.gov, identifier
NCT01462175. This compound is disclosed in US Pub 2010/0152190 A1.
To the extent necessary, this patent publication is herein
incorporated by reference.
[0005] Applicants have discovered that Compound A is especially
effective, and best tolerated, in human cancer therapy when
administered in the specific doses and pursuant to the specific
protocols herein described.
SUMMARY OF THE INVENTION
[0006] The present invention relates to a method of treating a
patient suffering with cancer, in particular colon, breast,
prostate, head and neck, lung or kidney cancer, leukemia, melanoma,
soft tissue sarcoma or osteosarcoma, comprising administering to
the patient Compound A in an amount of from about 800 to about 3000
mg/day, or from about 1000 to about 2500 mg/day, or from about 1250
to about 1800 mg/day, for an administration period of up to about 7
days, preferably up to about 5 days, on days 1-7, or preferably
days 1-5, of a 28 day treatment cycle, followed by a rest period of
from about 21 to about 23 days, preferably up to about 23 days.
[0007] The present invention further relates to a method of
treating a patient suffering from cancer, in particular colon,
breast, prostate, head and neck, lung or kidney cancer, leukemia,
melanoma, or soft tissue sarcoma, osteosarcoma comprising
administering to the patient a low concentration of Compound A in
an amount of from about 400 to about 1600 mg/day for an
administration period of up to about 7 days, preferably up to about
5 days, on days 1-7, or preferably days 1-5, of a 28 day treatment
cycle, followed by a rest period of from about 21 to about 23 days,
preferably about 23 days.
[0008] The present invention further relates to a method of
treating a patient suffering from cancer comprising administering
to the patient Compound A in an amount from about 400 mg/day to
about 1200 mg/day for up to about 7 days preferably 5 days,
followed by a rest period of about 21-23 days, of a 28 day
treatment cycle.
[0009] The present invention further relates to a method of
treating a patient suffering from cancer comprising administering
to the patient Compound A in an amount of from about 400-1000
mg/day for about 3 days followed by a rest period of about 25 days
of a 28 day treatment cycle.
[0010] The present invention further relates to a method of
treating a patient suffering from cancer comprising administering
to the patient Compound A in an amount of from 1600-2400 mg/once
weekly.times.3 weeks followed by about 13 days of rest, said
administration starting on the first day of a 28 day treatment
cycle.
[0011] The present invention further relates to a method of
treating a patient suffering from cancer, comprising administering
to said patient a pharmaceutical composition containing as an
active ingredient Compound A at about 500 mg/day total, daily for
up to about 5 days, followed by a rest period of about 23 days,
said administration starting on the first day of a 28 day treatment
cycle.
[0012] The present invention further relates to a method of
treating a patient suffering from leukemia, comprising
administering to said patient a pharmaceutical composition
containing as an active ingredient Compound A at about 1200 mg
total daily, for up to about 5 days, followed by a rest period of
about 23 days, said administration starting on the first day of a
28 day treatment cycle.
BRIEF DESCRIPTION OF THE DRAWINGS
[0013] FIG. 1 illustrates the antitumor activity, as demonstrated
by the change in mean tumor volume over time, of Compound A
monotherapy for a number of different dosing schedules, including a
continuous 5 day dosing schedule.
[0014] FIG. 2 shows the increased lifespan of mice treated with
Compound A for the different dosing schedules also reflected in
FIG. 1.
[0015] FIG. 3 illustrates the efficacy of Compound A for treating
leukemia using the 5 day dosing schedule.
DETAILED DESCRIPTION OF THE INVENTION
[0016] "Tumor control" means that the perpendicular diameters of
measurable lesions have not increased by 25% or more from the last
measurement. See, e.g., World Health Organization ("WHO") Handbook
for Reporting Results of Cancer Treatment, Geneva (1979). The
determination of tumor control or shrinkage (also referred to as
"regression") is made by known methods. For example, by evaluation
of patient symptoms, physical examination, X-ray, MRI or CAT scan
or other commonly accepted evaluation modalities.
[0017] The present invention relates to a method of treating a
patient suffering with cancer, in particular colon, breast,
prostate, head and neck, lung or kidney cancer, leukemia, melanoma,
soft tissue sarcoma or osteosarcoma, comprising administering to
the patient Compound A in an amount of from about 400 to about 1600
mg/day, or from about 400 to about 1200 mg/day, for an
administration period of up to about 7 days, preferably up to about
5 days, on days 1-7, or preferably days 1-5, of a 28 day treatment
cycle, followed by a rest period of from about 21 to about 23 days,
preferably up to about 23 days. The course of a preferred cycle is
about 28 days, though cycles anywhere between about 14 and about 28
days are contemplated. This treatment cycle is repeated for as long
as the tumor remains under control and the regimen is clinically
tolerated.
[0018] Dosages of Compound A can be applied either as a body
surface area ("BSA") adapted dose (mg/m.sup.2/day) or following
flat dosing (mg/day). Compound A may be administered as a single
dose daily or divided into multiple daily doses.
[0019] A patient's body measurement in square meters ("m.sup.2")
typically ranges from about 1.4 m.sup.2 to about 2.2 m.sup.2. Thus,
the total amount of Compound A to be delivered in a treatment cycle
(mg) using a BSA adapted dose would be calculated as follows: [Dose
intensity (mg/m.sup.2/week)].times.[BSA(m.sup.2)].times.[number of
weeks in treatment cycle].
[0020] In an embodiment, Compound A is administered daily for about
5 days, on days 1-5 of a treatment cycle, followed by a rest period
of 23 days ("5+/23-"). The 5+/23-treatment schedule is expected to
be superior to interim schedules or to longer schedules as
currently on-going Phase I studies indicate that in solid tumors,
maximal apoptosis occurs only after about 48 hours of continuous
exposure and longer schedules seem to present occurrence of delayed
thrombocytopenia ("TCP"). Thus, a 3-5 daily treatment schedule is
expected to provide the best benefit ratio taking into
consideration efficacy and toxicity.
[0021] In the present invention, the patient is administered a
lower dose of Compound A in an amount from about 400 to about 1600
mg/day for an administration period of up to about 7 days
preferably about 5 days, on days 1-7, or preferably days 1-5, of a
28 day treatment cycle. It has been unexpectedly found that it is
at least as effective and safer to treat humans with lower and less
frequent doses of Compound A than administering frequent doses at
lower concentrations or larger doses at intermittent periods. It
has been found that the dosing regimens are effective in eliciting
biological response from patients and better tolerated in terms of
safety.
[0022] Compound A is administered daily, either once or twice (bid)
daily. The compound is administered to the patient in an oral unit
dosage form, most preferably in tablet form.
[0023] Preferably, the 5 day treatment schedule is repeated every
twenty-eight days, or as soon as permitted by recovery from
toxicity, for so long as the tumor is under control or regressing
and the patient tolerates the regimen. Preferably, these treatment
cycles are repeated for up to about 12 cycles.
[0024] In an embodiment, Compound A is administered daily in an
amount from about 400 to about 1600 mg/day for up to about 5 days
on days 1-5 of a 28 day cycle.
[0025] In another embodiment, Compound A is administered in an
amount from about 400 mg/day to about 1200 mg/day, daily, for about
5 days, followed by a rest period of up to about 23 days, said
administration starting on the first day of a 28 day treatment
cycle.
[0026] In another embodiment, Compound A is administered daily in
an amount from about 400-1000 mg/day for up to about 3 days
followed by 25 days of rest of a 28 day cycle.
[0027] In another embodiment, Compound A is administered once
weekly in an amount from 1600-2400 mg/week for 3 weeks followed by
about 13 days of rest of a 28 day schedule.
[0028] In another embodiment Compound A is administered to treat
advance solid tumors including colon, breast, prostate, head, neck,
lung, kidney, melanoma, soft tissue sarcoma or osteosarcoma at
about 500 mg/day total, daily for up to about 5 days, followed by a
rest period of about 23 days, said administration starting on the
first day of a 28 day treatment cycle.
[0029] In another embodiment Compound A is administered to treat
leukemia at about 600 mg/day two times a day, for up to about 5
days, followed by a rest period of about 23 days, said
administration starting on the first day of a 28 day treatment
cycle.
[0030] The present invention may be exemplified by controlled
preclinical animal studies as shown in the Examples below, which
illustrates the invention without limitation.
[0031] The present invention is further exemplified by clinical
studies in humans as shown in the examples below which illustrate
the invention without limitation.
EXAMPLES
[0032] The superiority of the 5 day, 3 day and weekly dosing
regimen of the present invention on solid tumors is demonstrated by
the following experiments.
[0033] Abbreviations used herein are as follows:
TABLE-US-00001 x times po orally bid twice daily wk week qd once
daily qdx5 once daily for five days qweekly or 1x/wk once a week
BWL body weight loss SD standard deviation
Toxicity
[0034] In the examples below, weight loss was graphically
represented as percent change in mean group body weight, using the
formula: ((W-W.sub.0)/W.sub.0).times.100, where `W` represents mean
body weight of the treated group at a particular day, and `W.sub.0`
represents mean body weight of the same treated group at initiation
of treatment. Maximum weight loss was also represented using the
above formula, and indicated the maximum percent body weight loss
that was observed at any time during the entire experiment for a
particular group. Toxicity is defined as .gtoreq.20% of mice in a
given group demonstrating .gtoreq.20% body weight loss and/or
death.
Tumor Growth Inhibition (TGI) and Assessment of Survival/Increase
in Life Span (ILS)
[0035] Efficacy data was graphically represented as the mean tumor
volume.+-.standard error of the mean (SEM). In addition, tumor
volumes of treated groups were presented as percentages of tumor
volumes of the control groups (% T/C), using the formula:
100.times.((T-T.sub.0)/(C-C.sub.0)), where T represented mean tumor
volume of a treated group on a specific day during the experiment,
T.sub.0 represented mean tumor volume of the same treated group on
the first day of treatment; C represented mean tumor volume of a
control group on the specific day during the experiment, and
C.sub.0 represented mean tumor volume of the same treated group on
the first day of treatment.
[0036] Tumor volume (in cubic millimeters) was calculated using the
ellipsoid formula: (D.times.(d.sup.2))/2, where "D" represents the
large diameter of the tumor and "d" represents the small diameter.
In some cases, tumor regression and/or percent change in tumor
volume was calculated using the formula:
((T-T.sub.0)/T.sub.0).times.100, where `T` represents mean tumor
volume of the treated group at a particular day, and `T.sub.0`
represents mean tumor volume of the same treated group at
initiation of treatment.
[0037] Statistical analysis was determined by the rank sum test and
One Way Anova and a post-hoc Bonferroni t-test (SigmaStat, version
2.0, Jandel Scientific, San Francisco, Calif., USA). Differences
between groups were considered to be significant when the
probability value (p) was .ltoreq.0.05.
[0038] For survival assessment, the percent of increased life space
(ILS) was calculated as: 100.times.[(median survival day of treated
group-median survival day of control group)/median survival day of
control group]. Median survival was determined utilizing Kaplan.
Meier survival analysis. Survival in treated groups was
statistically compared with the vehicle group and survival
comparisons were done between groups using the log-rank test (Graph
Pad Prism, La Jolla, Calif., USA). Differences between groups were
considered significant when the probability value (p) was
.ltoreq.0.05.
Example 1
[0039] The antitumor activity of Compound A in the human
osteosarcoma cancer xenograft model SJSA1 in immunocompromized mice
using a variety of different schedules was assessed.
A. Test Compound A
[0040] Compound A was formulated as an amorphous solid dispersion
micro-bulk precipitate (MBP) powder containing 30% drug substance
and 70% HPMC-AS polymer was reconstituted immediately before
administration as a suspension in Klucel/Tween, and remaining
suspension was discarded after dosing. All dose levels are reported
as the actual dosage of Compound A rather than including drug plus
polymer.
B. In Vivo Assays
Animals
[0041] Female athymic Crl:NU-Foxnlnu mice (10/group), obtained from
Charles River Laboratories (Wilmington, Del.) were utilized when
they were approximately 10-12 weeks of age and weighed 23-25 g. The
health of the mice was assessed daily by gross observation and
analyses of blood samples taken from sentinel animals housed on
shared shelf racks. All animals were allowed to acclimate and
recover from any shipping-related stress for a minimum of 72 hours
prior to experimental use. Autoclaved water and irradiated food
(5058-ms Pico Lab mouse chow, Purina Mills, Richmond, Ind.) were
provided ad libitum, and the animals were maintained on a 12 hour
light and dark cycle. Cages, bedding and water bottles were
autoclaved before use and changed weekly. All animal experiments
were conducted in accordance with the Guide for the Care and Use of
Laboratory Animals, local regulations, and protocols approved by
the Roche Animal Care and Use Committee in an AAALAC accredited
facility.
Tumors
[0042] SJSA cells (ATCC) were maintained in RPMI 1640+10% (v/v)
heat-inactivated FBS+1% (v/v) 200 nM L-glutamine. Each mouse
received 5.times.10.sup.6 cells in a 1:1 mixture of phosphate
buffered saline and Matrigel in a total volume of 0.2 ml. Cells
were implanted subcutaneously in the right flank using a 1 cc
syringe and a 26 gauge needle.
Study Design:
[0043] The doses selected for Compound A and schedules utilized in
this study are shown in Table 1 below.
TABLE-US-00002 TABLE 1 Study Design Tumor Model Treatment Groups
SJSA 1. Vehicle qd po 2. Compound A 7.5 mg/kg qd po 3. Compound A
15 mg/kg qd po 4. Compound A 30 mg/kg qd po 5. Compound A 20 mg/kg
20 days qd po, 8 days off 6. Compound A 50 mg/kg 1x/week po 7.
Compound A 100 mg/kg 1x/week po 8. Compound A 200 mg/kg (given as
two 100 mg/kg doses 8 hours apart (bid)), 1x/week po 9. Compound A
50 mg/kg 4 days qd po, 10 days off x 2 cycles 10. Compound A 50
mg/kg 2 days qd po, 5 days off x 4 cycles 11. Compound A 100 mg/kg
2 days qd po, 5 days off x 4 cycles 12. Compound A 80 mg/kg 5 days
qd po, 23 days off 13. Compound A 100 mg/kg 2 days qd po, 12 days
off x 2 cycles
Treatment
[0044] Compound A was administered orally (po) using a 1 cc syringe
and 18-gauge gavage needle (0.2 ml/animal). Treatment duration was
2-4 weeks. Dates of tumor implant, treatment initiation (study
start date), and termination of treatment (study end date) can be
found in Table 6 below. The starting tumor volume for this study
was about 220 mm.sup.3. Tumor volumes and animal body weights were
measured three times per week and animals were monitored for
clinical signs daily.
[0045] The results of this experiment are summarized Tables 1-3
below and FIGS. 1 and 2. As can be seen, the 5 day treatment
schedule yielded the greatest percent increase in life span (% ILS)
as well as high percent tumor growth inhibition (% TGI) with
reasonable toxicity. FIG. 1 also shows good growth inhibitory
activity of the 5 day on/23 day off treatment schedule.
TABLE-US-00003 TABLE 2 Toxicity Summary % Change in Body Weight at
end of Maximum Maximum # of animals Group Frequency Study Day 29 %
Weight loss % Weight gain .gtoreq.20% BWL Mortality Vehicle QD 13.0
-1.2 13.0 0 0 Compound A 1x/wk 9.1 4.2 9.1 0 0 100 mg/kg Compound A
1x/wk 6.3 1.9 6.3 0 0 200 mg/kg (Two 100 mg/kg doses, 8 hr apart
Compound A 2 on/5 off x 4, QD 7.1 -0.8 7.1 0 0 50 mg/kg Compound A
5 on/23 off, QD 8.0 0.3 8.0 0 0 80 mg/kg Compound A 20 on/8 off, QD
1.2 -3.9 1.2 0 0 20 mg/kg Compound A 2 on/12 off x 2, QD 0.9 -0.6
1.8 0 0 100 mg/kg Compound A 4 on/10 off x 2, QD 1.2 -1.1 1.2 0 0
50 mg/kg Compound A QD 5.9 -2.2 5.9 0 0 15 mg/kg Compound A 2 on/5
off x 4, QD 1.3 -2.8 1.3 0 0 100 mg/kg Compound A QD 1.3 -0.2 1.3 0
0 30 mg/kg Compound A 1 x/wk 6.6 -0.3 6.6 0 0 50 mg/kg Compound A
QD 9.0 -0.3 9.0 0 0 7.5 mg/kg
TABLE-US-00004 TABLE 3 Efficacy Summary (left side) Mean Tumor Mean
Tumor Group Vehicle (mm3) Start Volume (mm3) or Compound A
Frequency Study Day: 11 SEM SD End Study Day: 32 SD SEM Vehicle QD
215.03 .+-.19.00 .+-.60.08 4696.49 .+-.785.28 .+-.296.91 50 mg/kg
1x/week 275.41 .+-.22.66 .+-.71.65 22.66 .+-.1103.00 .+-.348.80 7.5
mg/kg QD 240.88 .+-.18.01 .+-.56.95 18.01 .+-.956.45 .+-.302.46 100
mg/kg 1 x/week 193.61 .+-.9.67 .+-.30.57 474.73 .+-.273.78
.+-.86.58 15 mg/kg QD 232.37 .+-.16.42 .+-.51.93 16.42 .+-.872.83
.+-.276.01 50 mg/kg 2 on/5 off x 4, QD 203.43 .+-.18.78 .+-.59.39
257.29 .+-.102.12 .+-.32.29 80 mg/kg 5 on/23 off, QD 197.38
.+-.12.80 .+-.40.48 128.05 .+-.84.89 .+-.26.84 20 mg/kg 20 on/8
off, QD 207.20 .+-.16.97 .+-.53.67 315.19 .+-.277.51 .+-.87.76 100
mg/kg 2 on/12 off x 2, QD 201.40 .+-.9.86 .+-.31.18 179.88
.+-.154.02 .+-.48.71 50 mg/kg 4 on/10 off x 2, QD 213.61 .+-.12.09
.+-.38.23 244.70 .+-.240.07 .+-.75.92 100 mg/kg 2 on/5 off x 4, QD
190.78 .+-.25.68 .+-.81.22 25.68 .+-.15.82 .+-.5.00 30 mg/kg QD
250.86 .+-.19.35 .+-.61.19 19.35 .+-.159.01 .+-.50.28 100 mg/kg 200
mg/kg 224.88 .+-.12.02 .+-.38.02 158.95 .+-.68.86 .+-.21.78 (Two
100 mg/kg doses, 8 hr apart) x 1x Efficacy Summary Continued (right
side) % T/C End % Inhibition P value Average % Animals % Increased
p Value of Study end of study end of study Regression Partial Full
per Life versus Day: 32 Day: 32 Day: 32 per Group Regression
Regression Group Span Vehicle -- -- -- -- 0 0 7 -- -- 43 57
<0.001 -- 0 0 10 23 0.0036 34 66 <0.001 -- 0 0 10 23 0.0012 6
94 <0.001 -- 1 0 10 77 <0.0001 21 79 <0.001 -- 0 0 10 62
<0.0001 1 99 <0.001 -- 3 0 10 119 <0.0001 -2 regression
<0.001 35 6 2 10 127 <0.0001 2 98 <0.001 -- 5 0 10 77
<0.0001 0 regression <0.001 11 7 0 10 119 <0.0001 1 99
<0.001 -- 6 0 10 112 <0.0001 -2 regression <0.001 47 9 0
10 188 <0.0001 -1 regression <0.001 13 7 0 10 127 <0.0001
-1 regression <0.001 29 7 0 10 162 <0.0001
TABLE-US-00005 TABLE 4 Survival Summary 50% 50% Treatment Vehicle %
p Group Days days ILS value Vehicle QD -- -- -- -- Compound A 1
x/wk 46 26 77 <0.0001 100 mg/kg Compound A Two 100 mg/kg 68 26
162 <0.0001 200 mg/kg doses, 8 hr apart 1 x/wk Compound A 2 on/5
off x 4, 57 26 119 <0.0001 50 mg/kg QD Compound A 5 on/23 off,
59 26 127 <0.0001 80 mg/kg QD Compound A 20 on/8 off, 46 26 77
<0.0001 20 mg/kg QD Compound A 2 on/12 off x 2, 57 26 119
<0.0001 100 mg/kg QD Compound A 4 on/10 off x 2, 55 26 112
<0.0001 50 mg/kg QD Compound A QD 42 26 62 <0.0001 15 mg/kg
Compound A 2 on/5 off x 4, 75 26 188 <0.0001 100 mg/kg QD
Compound A QD 59 26 127 <0.0001 30 mg/kg Compound A 1 x/wk 32 26
23 0.0036 50 mg/kg Compound A QD 32 26 23 0.0012 7.5 mg/kg
Example 2
[0046] The antitumor activity of Compound A in the human acute
myeloid leukemia xenograft model MOLM-13 in immunocompromized mice
using an oral qdx5 schedule was assessed.
A. Test Compound A
[0047] Compound A was formulated as an amorphous solid dispersion
micro-bulk precipitate (MBP) powder containing 30% drug substance
and 70% HPMC-AS polymer was reconstituted immediately before
administration as a suspension in Klucel/Tween, and remaining
suspension was discarded after dosing. All dose levels are reported
as the actual dosage of Compound A rather than including drug plus
polymer.
B: In Vivo Assays
Animals
[0048] Female SCID:beige mice (10/group), obtained from Charles
River Laboratories (Wilmington, Del.) were utilized when they were
approximately 8-10 weeks of age and weighed 21-23 g. The health of
the mice was assessed daily by gross observation and analyses of
blood samples taken from sentinel animals housed on shared shelf
racks. All animals were allowed to acclimate and recover from any
shipping-related stress for a minimum of 72 hours prior to
experimental use. Autoclaved water and irradiated food (5058-ms
Pico Lab mouse chow, Purina Mills, Richmond, Ind.) were provided ad
libitum, and the animals were maintained on a 12 hour light and
dark cycle. Cages, bedding and water bottles were autoclaved before
use and changed weekly. All animal experiments were conducted in
accordance with the Guide for the Care and Use of Laboratory
Animals, local regulations, and protocols approved by the Roche
Animal Care and Use Committee in an AAALAC accredited facility.
Tumors
[0049] MOLM-13 cells (ATCC) were maintained in RPMI 1640 with
L-glutamine (2 mM) media (GIBCO/Invitrogen, Carlsbad, Calif.)
supplemented with 10% heat-inactivated Fetal Bovine Serum (HI-FBS;
GIBCO/Invitrogen, Carlsbad, Calif.), and 1% 100 mM sodium pyruvate.
Each mouse received 1.times.10.sup.6 cells suspended in Phosphate
Buffered Saline (PBS) intravenously inoculated via the caudal tail
vein in a volume of 0.2 ml using a 1 cc syringe and a 26 gauge
needle.
Study Design:
[0050] The doses selected for Compound A and schedules utilized in
this study are shown in Table 5 below.
TABLE-US-00006 TABLE 5 Study Design Tumor Model Treatment Groups
MOLM- 1. Vehicle qdx5 po 13 2. Compound A 80 mg/kg 5 days qd po
(qdx5), 23 days off
Treatment
[0051] Compound A was administered orally (po) using a 1 cc syringe
and 18-gauge gavage needle (0.2 ml/animal). Treatment duration was
initiated on day 3 and was for 2 weeks Animal body weights were
measured three times per week and animals were monitored for
clinical signs daily.
[0052] The results of this experiment are summarized Tables 6-7
below and FIG. 3. The 5 day treatment schedule yielded a 37%
percent increase in life span (% ILS) in the absence of treatment
related toxicity.
TABLE-US-00007 TABLE 6 Toxicity Summary % Change in Body Weight at
end of Maximum Maximum # of animals Mortality Group Frequency Study
Day 17 % Weight loss % Weight gain .gtoreq.20% BWL (reason) Vehicle
QD -4.9 -6.0 -2.5 0 0 Compound A qdx5 -3.2 -5.5 -3.2 0 1 (dosing 80
mg/kg injury)
TABLE-US-00008 TABLE 7 Efficacy/Survival Summary 50% 50% Treatment
Vehicle Group Days days % ILS p value Vehicle qdx5 -- -- -- --
Compound A qdx5 28 21 37 <0.05 80 mg/kg
Example 3
A. Compound a Composition and Administration
[0053] Film-coated tablets containing Compound A at dosage
strengths of 10 mg, 50 mg, 200 mg, and 400 mg were developed. The
10 mg tablets contain hypromellose acetate succinate, mannitol,
microcrystalline cellulose, croscarmellose sodium, colloidal
silicon dioxide, and magnesium stearate. The film-coat consists of
hypromellose, titanium dioxide, talc, triacetin, iron oxide yellow,
and iron oxide red. The 50 mg, 200 mg, and 400 mg film coated
tablets contain the excipients hypromellose acetate succinate,
croscarmellose sodium, colloidal silicon dioxide, and magnesium
stearate. The film-coat consists of polyvinyl alcohol, titanium
dioxide, polyethylene glycol, talc, iron oxide yellow, iron oxide
red, and iron oxide black.
[0054] The dosage amount can vary and will, of course, have to be
adjusted to the individual requirements in each particular case. In
the case of oral administration, the dosage can vary from about 100
mg to about 3000 mg per day of Compound A. The daily dosage can be
administered as single dose or in divided doses.
B. Clinical Design
[0055] By way of nonlimiting examples, patients with advanced solid
tumors including colorectal cancer, prostate cancer, lung cancer,
kidney cancer and breast cancer were administered Compound A
formulated as described above under the following schedules and
dosage ranges: (i) 1600-2400 mg total weekly dose (once
weekly.times.3 followed by about 13 days of rest), (ii) 1000 mg
total daily dose (daily.times.3 followed by 25 days of rest), and
(iii) 500-800 mg total daily dose (daily.times.5 followed by 23
days of rest). Tumor biopsies were collected at baseline and cycle
1, day 5.+-.2. The pharmacological effects of Compound A was
assessed by measuring biomarker changes between baseline and post
treatment.
C. Biological Assays
[0056] Biological assay results of the administration of Compound A
on patients according to the clinical design schedule and dosage
ranges described above are set forth in Tables 8-11. Activity from
biomarker assays shows direct p53 activation (Table 5), and
anti-proliferative effects Ki67 (Table 6), TUNEL (Table 7), and
FLT-PET (Table 8).
TABLE-US-00009 TABLE 8 MDM2 mRNA Assay MDM2 mRNA concentrations
from human tumor tissue were assessed by RT PCR. Treatment with
Compound A under the doses below increased the levels of p53
leading to an increase in the levels of MDM2 mRNA expression in
patient's tumors. This elevation in the levels of MDM2 mRNA is
consistent with activation of the p53 pathway by Compound A. MDM2
mRNA Median Average Cohort and Schedule Patient FCBL FCBL FCBL
BM-A1 800 mg bid 1501 1.0 1.1 1.2 Qwk x 3 1701 1.4 1702 1.1 BM-A2
1200 mg bid 1602 1.3 1.3 1.4 Qwk x 3 1703 1.0 1603 1.0 1201 1.8
1503 2.1 BM-B1 400 mg bid 2701 5.4 8.7 8.5 x5 d 2502 3.7 2702 12.1
2404 12.9 BM-B2 500 mg bid 2405 0.2 1.3 5.1 x 3 d 2504 0.7 2203 1.3
2204 3.8 2505 19.6 BM-B3 500 mg qd 2703 1.5 2.0 2.4 x 5 d 2506 2.5
2406 4.2 2705 1.2
TABLE-US-00010 TABLE 9 Ki-67 Assay The Ki-67 protein is a cellular
marker for proliferation. It is strictly associated with cell
proliferation and can be measured in tissue sections via
immunohistochemistry to determine degree of proliferation. Ki-67
protein is present during all active phases of the cell cycle (G1,
S, G2, and mitosis), but is absent from resting cells (G0).
Decrease levels of tumor proliferation in patients were observed
via Ki67 after treatment with Compound A under the doses and
schedules below. This decrease in proliferation is in response to
the p53 activation induced by Compound A. Ki-67 IHC Median Average
Cohort and Schedule Patient % CBL % CBL % CBL BM-A1 800 mg bid 1501
14 -8.5 -8.5 Qwk x 3 1701 -31 BM-A2 1200 mg bid 1602 -33 -33.0
-27.8 Qwk x 3 1703 -57 1603 30 1201 -5 1503 -75 BM-B1 400 mg bid
2701 -97 -13.0 -17.7 x5 d 2502 -13 2404 57 BM-B2 500 mg bid 2503 3
-69.8 -53.9 x 3 d 2504 -70 2204 -70 2505 -79 BM-B3 500 mg qd 2703
-67 -66.5 -57.1 x 5 d 2506 -96 2406 -8
TABLE-US-00011 TABLE 10 TUNEL Assay Tumor biopsy cells were
analyzed via TUNEL (terminal deoxynucleotidyl transferase dUTP nick
end labeling) to determine cell apoptosis. TUNEL is a common method
for detecting DNA fragmentation that results from apoptotic
signaling cascades. The assay relies on the presence of nicks in
the DNA which can be identified by terminal deoxynucleotidyl
transferase or TdT, an enzyme that will catalyze the addition of
dUTPs that are secondarily labeled with a marker. Treatment with
Compound A lead to an increase in apoptosis and this is in response
to the p53 activation induced by Comound A. TUNEL + cells (mm2)
Median Average Cohort and Schedule Patient FCBL FCBL FCBL BM-A1 800
mg bid 1501 0.8 0.8 1.2 Qwk x 3 1701 0.8 1601 3,2 1702 0.1 BM-A2
1200 mg bid 1602 5.3 2.3 2.7 Qwk x 3 1703 2.3 1603 0.9 1201 1.6
1503 3.3 BM-B1 400 mg bid 2701 0.9 1.8 1.7 x5 d 2601 1.8 2502 1.8
2702 0.9 2404 3.2 BM-B2 500 mg bid 2405 0.5 0.9 1.7 x 3 d 2503 0.4
2504 0.6 2203 5.3 2204 1.2 2505 1.9 BM-B3 500 mg qd 2703 0.6 1.2
1.2 x 5 d 2506 1.7 2406 1.2
TABLE-US-00012 TABLE 11 FLT PET Assay FLT PET imaging scan was used
to analyze tumor proliferation in patients undergoing treatment
with Compound A under the doses and schedules set forth below. A
negative percent change from baseline is indicative of an
antiproliferative effect. This is in alignment with the Ki67 and
TUNEL results. By FLT PET, an antiproliferative effects was seen in
all doses tested in particular the qd x 3 days and qd x 5 day
schedules. FLT PET Median Average Cohort and Schedule Patient % CBL
Cycle 1 % CBL % CBL BM-A1 800 mg bid 1407 -19.0 -20.5 -17.0 Qwk x 3
1501 -11.7 1701 -28.5 1408 -21.9 1601 -29.5 1702 8.9 BM-A2 1200 mg
bid 1602 -5.0 3.2 -0.9 Qwk x 3 1703 21.1 1603 -31.1 1201 11.3 1503
-73.1 BM-B1 400 mg bid 2701 -82.5 -32.2 -43.4 x5 d 2501 -49.7 2502
-29.3 2702 -23.4 2404 -32.2 BM-B2 500 mg bid 2405 -34.9 -34.9 -39.2
x 3 d 2504 -62.4 2203 -24.7 2204 -19.4 2505 -54.5 BM-B3 500 mg qd
2703 -56.4 -31.0 -29.04 x 5 d 2704 -34.3 2205 -21.7 2506 -31 2406
-1.8
* * * * *