U.S. patent application number 10/633114 was filed with the patent office on 2004-02-05 for cancer treatment with epothilones.
Invention is credited to Cohen, Pamela, Litchman, Manuel, O'Reilly, Terence, Wartmann, Markus.
Application Number | 20040024033 10/633114 |
Document ID | / |
Family ID | 27517450 |
Filed Date | 2004-02-05 |
United States Patent
Application |
20040024033 |
Kind Code |
A1 |
O'Reilly, Terence ; et
al. |
February 5, 2004 |
Cancer treatment with epothilones
Abstract
The invention relates to the treatment of a proliferative
disease, especially according to certain treatment regimens, with
an epothilone, especially with epothilone A and more preferably
epothilone B; as well as to the treatment of certain cancers with
such an epothilone.
Inventors: |
O'Reilly, Terence; (Basel,
CH) ; Wartmann, Markus; (Riehen, CH) ;
Litchman, Manuel; (Teaneck, NJ) ; Cohen, Pamela;
(Tenafly, NJ) |
Correspondence
Address: |
THOMAS HOXIE
NOVARTIS, CORPORATE INTELLECTUAL PROPERTY
ONE HEALTH PLAZA 430/2
EAST HANOVER
NJ
07936-1080
US
|
Family ID: |
27517450 |
Appl. No.: |
10/633114 |
Filed: |
August 1, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10633114 |
Aug 1, 2003 |
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09925119 |
Aug 9, 2001 |
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6635666 |
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09925119 |
Aug 9, 2001 |
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09233993 |
Jan 19, 1999 |
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6302838 |
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Current U.S.
Class: |
514/365 |
Current CPC
Class: |
A61P 35/00 20180101;
A61K 31/425 20130101 |
Class at
Publication: |
514/365 |
International
Class: |
A61K 031/427 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 25, 1998 |
GB |
9803907.6 |
Feb 25, 1998 |
GB |
9803905.0 |
Mar 19, 1998 |
GB |
9805936.3 |
Mar 19, 1998 |
GB |
9805937.1 |
Claims
What we claim is:
1. A method for treating a proliferative disease, said method
comprising the step of administering, in at least one treatment, a
therapeutically effective amount of an epothilone, together with a
pharmaceutically acceptable carrier, to a warm-blooded animal in
need of such treatment.
2. The method of claim 1 where the epothilone is epothilone B.
3. The method of claim 2 where epothilone B is administered in more
than one treatment with an interval of about one week to about six
weeks between treatments.
4. The method of claim 2 where epothilone B is administered in a
dose to humans that is calculated according to the formula (I)
single dose(mg/m.sup.2)=(0.1 to y).times.N (I) where N is the
number of weeks between treatments and y is 6.
5. The method of claim 2 where epothilone B is administered in a
dose to humans that is calculated according to the formula II
single dose(mg/m.sup.2)=(0.1 to 2.5).times.N (II).
6. The method of claim 4 where epothilone B is administered weekly
in a dose that is between about 0.1 and about 6 mg/m.sup.2.
7. The method of claim 4 where epothilone B is administered every 6
to 8 days in a dose that is between about 0.1 and about 5
mg/m.sup.2.
8. The method of claim 4 where epothilone B is administered every 6
to 8 days in a dose that is between about 0.1 and about 3
mg/m.sup.2.
9. The method of claim 4 where epothilone B is administered every
week in a dose between about 0.3 and about 1 mg/m.sup.2.
10. The method of claim 4 where epothilone B is administered every
third week in a dose that is between about 0.3 and about 18
mg/m.sup.2.
11. The method of claim 4 where epothilone B is administered once
every 18 to 24 days three weeks in a dose that is between about 0.3
and about 12 mg/m.sup.2.
12. The method of claim 4 where epothilone B is administered every
18 to 24 days in a dose that is between about 0.3 and about 7.5
mg/m.sup.2.
13. The method of claim 4 where epothilone B is administered every
third week in a dose that is between about 1.0 and about 3.0
mg/m.sup.2.
14. The method of claim 4 where epothilone B is administered by
intravenous infusion.
15. The method of claim 6 where the epothilone is administered by
intravenous infusion.
16. The method of claim 15 where each infusion takes place during
about 5 to about 30 min.
17. The method of claim 10 where epothilone B is administered by
intravenous infusion.
18. The method of claim 17 where the infusion takes place during
about 5 to about 30 min.
19. The method of claim 1 wherein the proliferative disease is
refractory to treatment with one or more chemotherapeutics other
than an epothilone, where epothilone B is administered to a human
in need of such treatment in a dose that is appropriate for the
treatment of said disease.
20. The method of claim 19 where the refractory tumor to be treated
is selected from the group consisting of lung, colorectal,
prostate, br ast or epidermoid head or neck tumors.
21. The method of claim 19, where the epothilone B is administered
in more than on treatment with an interval between treatments of
about one week to about six weeks after the preceding treatment,
respectively.
22. The method of claim 19 where epothilone B is administered by
intravenous infusion in a dose to humans that is calculated
according to the formula (I) single dose(mg/m.sup.2)=(0.1 to
y).times.N (I) where N is the number of weeks between treatments
which lies between about 1 and about 3 weeks so that N also is
between about 1 and about 3, and y is 6.
23. The method of claim 22 where epothilone B is administered
weekly in a dose that is between about 0.1 and about 6
mg/m.sup.2.
24. The method of claim 22 where epothilone B is administered every
6 to 8 days in a dose that is between about 0.1 and about 5
mg/m.sup.2.
25. The method of claim 22 where epothilone B is administered every
6 to 8 days in a dose that is between about 0.1 and about 3
mg/m.sup.2.
26. The method of claim 22 where epothilone B is administered
weekly in a dose between about 0.3 and about 1 mg/m.sup.2.
27. The method of claim 22 where epothilone B is administered every
third week in a dose that is between about 0.3 and about 18
mg/m.sup.2.
28. The method of claim 22 where epothilone B is administered every
18 to 24 days in a dose that is between about 0.3 and about 12
mg/m.sup.2.
29. The method of claim 22 where epothilone B is administered every
18 to 24 days in a dose that is between about 0.3 and about 7.5
mg/m.sup.2.
30. The method of claim 22 where epothilone B is administered v ry
third week in a dose that is between about 1.0 and about 3.0
mg/M.sup.2.
31. The method of claim 22 where the infusion takes place during
about 5 to about 30 min.
32. The method of claim 22 wherein the tumor to be treated is a
colorectal tumor that is refractory to at least one member of the
taxane class of anti-cancer agents.
33. The method according to claim 32 wherein the colorectal tumor
to be treated is in addition refractory to at least one other
standard chemotherapeutic.
34. The method of claim 33 where the tumor to be treated is a
colorectal tumor that is refractory to TAXOL and 5-fluorouracil
treatment.
35. The method of claim 22 where the tumor to be treated is a
prostate tumor, and/or any metastasis, refractory to hormone
treatment.
36. The method of claim 22 where the tumor to be treated is an
epidermoid head or neck tumor that is refractory to treatment with
at least one other chemotherapeutic.
37. The method of claim 36 where the epidermoid head or neck tumor
is refractory to treatment with TAXOL.
38. The method of claim 22, where the tumor to be treated is a lung
tumor that is refractory to treatment with at least one other
chemotherapeutic.
39. The method of claim 38 where the tumor to be treated is a
non-small cell lung cancer.
40. The method of claim 39 where the non-small cell lung cancer is
refractory to treatment with TAXOL.
41. The method of claim 22 where the tumor to be treated is a
breast tumor.
42. The method of claim 31 wherein the tumor to be treated is a
colorectal tumor that is refractory to at least one member of the
taxane class of anti-cancer agents.
43. The method of claim 42 wherein the tumor to be treated is a
colorectal tumor that is refractory to standard chemotherapy.
44. The method of claim 43 where the tumor to be treated is a
colorectal tumor that is refractory to TAXOL and 5-fluorouracil
treatment.
45. The method of claim 31 where the tumor to be treated is a
prostate tumor, and/or any metastasis thereof, wherein the tumor
and/or its metastasis is refractory to hormone treatment.
46. The method of claim 31 where the tumor to be treated is an
epidermoid head or neck tumor refractory to treatment with at least
one other chemotherapeutic due to multi-drug resistance.
47. The method of claim 46 where the epidermoid head or neck tumor
is refractory to treatment with TAXOL.
48. The method of claim 31, where the tumor to be treated is a lung
tumor that is refractory to treatment with at least one other
chemotherapeutic.
49. The method of claim 48 wh r the tumor to be treated is a
non-small cell lung cancer.
50. The method of claim 49 where the non-small cell lung cancer is
refractory to treatment with TAXOL.RTM..
51. The method of claim 31 where the tumor to be treated is a
breast tumor.
52. The method of claim 2 where the proliferative disease to be
treated is selected from the group consisting of a colorectal
tumor, a tumor of the genitourinary tract, an epidermoid tumor, a
lung tumor and a breast tumor.
53. The method of claim 52 where the proliferative disease to be
treated is a colorectal tumor that is refractory to at least one
member of the taxane class of anti/cancer agents and/or to standard
chemotherapy.
54. The method of claim 52 where the proliferative disease to be
treated is a prostate tumor.
55. The method of claim 54 where the prostate tumor is refractory
to hormone treatment.
56. The method of claim 52 where the proliferative disease is an
epidermoid head or neck tumor.
57. The method of claim 56 where the head or neck tumor is
multidrug-resistant.
58. The method according to claim 52 where the proliferative
disease is a non-small cell lung tumor.
59. The method of claim 58 where the non-small cell lung tumor is
refractory to treatment with a member of the taxane class of
anti-cancer agents.
60. The method of claim 52 where the proliferative disease is a
breast tumor.
61. The method of claim 60 where the breast tumor is refractory to
treatment with at least one member of the taxane class of
anti-cancer agents.
62. The method of claim 1 where the proliferative disease to be
treated is a multidrug resistant tumor.
63. The method of claim 1 where the proliferative disease to be
treated is selected from the group consisting of a melanoma,
ovarian cancer, pancreas cancer, neuroblastoma, head or neck
cancer, bladder cancer, renal cancer, brain cancer and gastric
cancer.
64. The method of claim 19, where epothilone B is administered in a
dose that is between about 1 and about 100% of the maximal
tolerated dose (MTD) to a human; and one or more further doses each
within 1% and 100% of the MTD are administered in at least one
additional treatment after an interval between the treatments of
one to three weeks.
65. The method of claim 64 where the dose to be administered is
between 25 and 100% of the MTD.
66. The method of claim 19, where epothilone B is administered
weekly to a human in a dose that is below 80% of the maximal
tolerable dose (MTD).
67. The method of claim 66, where the dose is below 50% of the
MTD.
68. The method of claim 1, further comprising the step of
administering (a) epothilone B in combination with (b) another
antitumor therapeutic, the combined treatment being so timed that
component (a) and component (b) are administered to a human in need
of such treatment in combination and in a quantity that is jointly
therapeutically effective against said proliferative disease.
69. The method of claim 2, where the proliferative disease is a
tumor that is refractory to the treatment with an anti-cancer agent
of the taxane class, said tumor being selected from the group
consisting of a colorectal, a prostatic, a pancreatic and a brain
tumor.
70. The method of claim 2 where the proliferative disease is a
multidrug resistant non-small cell lung carcinoma, a multidrug
resistant breast tumor, or a multidrug resistant epidermoid head
and neck tumor.
71. The use of an epothilone for the manufacture of a
pharmaceutical preparation for the treatment of a proliferative
disease, comprising admixing said epothilone with a
pharmaceutically acceptable carrier.
72. The use of claim 71, where the epothilone is epothilone B.
Description
SUMMARY OF THE INVENTION
[0001] The invention relates to the treatment of a proliferative
disease, especially according to certain treatment regimens using
an epothilone, especially epothilone B; preferably of a
gastrointestinal tumor, more preferably (1) a tumor of the colon
and/or the rectum (colorectal tumor), especially if it is
refractory to a (meaning at least one) representative of the taxane
class of anti-cancer agents, in particular TAXOL.RTM. (paclitaxel
in formulated form for clinical use), and/or at least one standard
treatment with an other chemotherapeutic, especially
5-fluorouracil; (2) a tumor of the genitourinary tract, more
preferably a tumor of the prostate, including primary and
metastatic tumors, especially if refractory to hormone treatment
("hormone refractory prostate cancer") and/or treatment with other
standard chemotherapeutics; (3) an epidermoid tumor, more
preferably an epidermoid head and neck tumor, most preferably a
mouth tumor; (4) a lung tumor, more preferably a non-small cell
lung tumor, especially any of these tumors that is refractory to
treatment with one or more other chemotherapeutics (especially due
to multidrug resistance), especially to treatment with a member of
the taxane class of anti-cancer agents, in particular TAXOL.RTM.;
or (5) a breast tumor, more preferably one that is multidrug
resistant, especially refractory to treatment with a member of the
taxane class of anti-cancer agents, in TAXOL.RTM.; relating
especially also to the treatment of a multidrug resistant lung
tumor (preferably a non-small cell lung tumor), a multidrug
resistant breast tumor, or a multidrug resistant epidermoid tumor,
or in a broader sense of the invention to a treatment schedule for
the treatment of an aforementioned or (in a broader sense of the
invention) any other tumor, especially if it is refractory to one
or more chemotherapeutics, especially multidrug resistant and/or
TAXOL.RTM. refractory), such as a melanoma, ovarian cancer,
pancreas cancer, neuroblastoma, head and neck cancer or bladder
cancer, or in a broader sense renal, brain or gastric cancer; by
administration of an epothilone as a cytotoxic agent, especially
epothilone B; the term "treatment" also encompassing (i) a method
of treatment for (=for treating of) said disease comprising
administration of said cytotoxic agent (preferably an epothilone,
especially epothilone B, in each case preferably together with a
pharmaceutically acceptable carrier) to a warm-blooded animal,
especially if in need of such treatment, in a therapeutically
effective amount, in at least one treatment; (ii) the use of said
cytotoxic agent, for the treatment of a proliferative disease;
(iii) the use of said cytotoxic agent for the manufacture of a
pharmaceutical preparation for the treatment of said proliferative
disease (comprising admixing said cytotoxic agent with a
pharmaceutically acceptable carrier); (iv) a pharmaceutical
preparation comprising a dose of said cytotoxic agent that is
appropriate for the treatment of said proliferative disease. The
invention is, in a preferred embodiment, directed to the treatment
of patients or patient groups where other treatments, especially
standard treatment with an other chemotherapeutic, especially
5-fluorouracil; or therapy with members of the taxane class of
anti-cancer agents, such as TAXOL.RTM., have failed.
BACKGROUND OF THE INVENTION
[0002] Cancer still represents a major unmet medical need. Initial
treatment of the disease is often surgery, radiation treatment or
the combination, but recurrent (metastatic) disease is common.
Chemotherapeutic treatments for most cancers are generally not
curative, but only delay disease progression. Commonly, tumors and
their metastases become refractory to chemotherapy, in an event
known as development of multidrug resistance. In many cases, tumors
are inherently resistant to some classes of chemotherapeutic agents
[see DeVita V. T., Principles of Cancer Management: Chemotherapy.
In: Cancer. Principles and Practice of Oncology. DeVita V. T. et al
(eds.), 5th edition, Lippincott-Raven, Philadelphia, N.Y. (1977),
pp. 333-347; or Cleton, F. J., Chemotherapy: general aspects. In:
Oxford Textbook of Oncology; Peckham, M., et al, Oxford University
Press, Oxford, N.Y., Tokyo (1995), Vol. 1, pp. 445-453]. This is,
for example, the case for lung tumors, especially non-small cell
lung carcinoma, or also for epidermoid tumors, like epidermoid head
and neck, especially mouth, tumors, or also for breast tumors.
Other mechanisms why tumors are not treatable (are refractory to
treatment) can be, for example, the presence of tubulin mutations
or glutathione mediated mechanisms.
[0003] Intestinal, especially colorectal, cancer defines a special
case of the unmet medical needs in cancer treatment. Initial
treatment of the disease is often surgery, radiation treatment or
the combination, but recurrent (metastatic) disease is common.
First-line chemotherapeutic treatments for recurrent colorectal
cancer include 5-fluorouracil. But this treatment provides at best
delay of disease progression as the tumors usually become
refractory to treatment. Chemotherapy of this refractory stage of
disease involves other classical cytotoxic agents, but are all
considered inadequate [see Cohen et al., Cancer of the colon. In:
Cancer. Principles and Practice of Oncology; DeVita et al. (eds.),
5th edition, Lippincott Raven. Philadelphia, N.Y. 1997, pp.
1144-1197; or Rowinsky, Ann. Rev. Med. 48, 353-74 (1997)]. Also for
cancer of the genitourinary tract, especially prostate cancer, a
further unmet medical need, initial treatment is as mentioned above
for colorectal cancer, showing similar problems. First-line
chemotherapeutic treatment for recurrent prostate cancer includes
anti-androgens, and the recurrence is frequently
androgen-dependent. But this treatment provides only delay of
disease progression as the tumors almost always become refractory
to anti-androgens within 6 months to 2 years (hormone-refractory
prostate tumors). Chemotherapy of this anti-androgen refractory
stage of diseases involves mitoxantrone or other classical
anticancer cytotoxic agents, but all are considered as inadequate
[see Oesterling et al., Cancer of the prostate. In: Cancer.
Principles and Practice of Oncology. DeVita, V. T., et al. (eds.),
5th edition, Lippincott-Raven, Philadelphia, N.Y. 1997, pp 1322-86;
Sternberg, Cancers of the genitourinary tract. In: Cavalli et al.
(eds.), Textbook of Medical Oncology; or Roth, B. J., Semin. Oncol.
23(6 Suppl. 14), 49-55 (1996)].
[0004] Among cytotoxic agents for the treatment of tumors,
TAXOL.RTM. (paclitaxel), a microtubule stabilizing agent, has
become a very important compound with a remarkable economic success
[see Rowinsky E. K., The development and clinical utility of the
taxane class of antimicrotubule chemotherapy agents; Ann. Rev. Med.
48, 353-374 (1997)].
[0005] However, TAXOL.RTM. has a number of disadvantages.
Especially its extremely low solubility in water represents a
severe problem. It has become necessary to administer TAXOL.RTM. in
a formulation with Cremophor EL.RTM. (polyoxyethylated castor oil;
BASF, Ludwigshafen, Germany) which has severe side effects, causing
inter alia allergic reactions that in one case even were reported
to have led to the death of a patient. More severely, certain tumor
types are known to be refractory to treatment with TAXOL.RTM. even
when the drug is administered as front-line therapy, or the tumors
develop resistance to TAXOL.RTM. after multiple cycles of
exposure.
[0006] Although the taxane class of antimicrotubule anti-cancer
agents has been hailed as the "perhaps most important addition to
the chemotherapeutic armamentarium against cancer over the past
several decades" [see Rowinsky E. K., Ann. Rev. Med. 48, 353-374
(1997)] and despite the commercial success of TAXOL.RTM., there
remain limitations to TAXOL.RTM.'s efficacy. TAXOL.RTM. treatment
is associated with a number of significant side effects and some
major classes of solid tumors, namely colon and prostate, are
poorly responsive to this compound (see Rowinsky E. K., loc. cit.).
Specifically, as a single agent, TAXOL.RTM. has been considered to
be poorly active clinically in colorectal, renal, prostatic,
pancreatic, gastric and brain cancers [see Rowinsky E. K., loc.
cit.; Bitton, R. J., et al., Drug Saf. 12, 196-208 (1995); or
Arbuck, S. G., et al., J. Natl. Cancer Inst. Monogr. 15, 11-24
(1993)]. For example, the effectiveness of TAXOL.RTM. can be
severely limited by acquired drug resistance mechanisms occurring
via various mechanisms, such as overexpression of
phosphoglycoproteins that function as drug efflux pumps.
[0007] Therefore, there exists an urgent need to find compounds and
appropriate dosing regimens with these compounds expand the
armamentarium of cancer treatment, especially in the majority of
cases where treatment with taxanes and other anticancer compounds
is not associated with long term survival.
[0008] The epothilones, especially epothilones A and B, represent a
new class of microtubule stabilizing cytotoxic agents (see Gerth,
K. et al., J. Antibiot. 49, 560-3 (1996); or Hoefle et al., DE 41
38 042), e.g. with the formulae: 1
[0009] wherein R is hydrogen (epothilone A) or methyl (epothilone
B).
[0010] These compounds have the following advantages:
[0011] (i) they show better water solubility than TAXOL.RTM. and
are thus more appropriate for formulation; and
[0012] (ii) they have, in cell culture experiments, been reported
to be active also against the proliferation of cells that, due to
the activity of the P-glycoprotein efflux pump which renders them
multidrug resistant, show resistance to treatment with other
chemotherapeutics, e.g. TAXOL [see Bollag, D. M., et al.,
"Epothilones, a new class of microtubule-stabilizing agents with a
Taxol-like mechanism of action", Cancer Research 55, 2325-33
(1995); and Bollag D. M., Exp. Opin. Invest. Drugs 6, 867-73
(1997)]; and
[0013] (iii) despite apparently sharing the same, or a sterically
proximal binding site on the microtubule, the epothilones have been
shown to be active against a TAXOL.RTM.-resistant ovarian carcinoma
cell line with an altered P-tubulin [see Kowalski, R. J., et al.,
J. Biol. Chem. 272(4), 2534-2541 (1997)1.
[0014] On the other hand, they are highly toxic and therefore their
usefulness in the treatment of cancer in vivo was considered
practically impossible [see, for example, PNAS 95, 9642-7 (1998)].
Therefore, the present invention shows in an unexpected way that
indeed dosage regimens may be found that allow, on the one hand, to
treat tumors with epothilones, especially epothilone B; and on the
other hand allow for the treatment of certain patient groups that
are unresponsive to other kinds of treatment, be it by multi-drug
resistance, as with taxane, e.g. TAXOL.RTM., refractoriness due to
multidrug resistance, and/or any other mechanism.
[0015] The present invention has the goal to present for the first
time in vivo regimens for a useful treatment with epothilones,
preferably epothilone A or especially epothilone B, that allow for
the treatment of a tumor disease, e.g. a melanoma, ovarian cancer,
pancreas cancer, neuroblastoma, head and neck cancer, bladder
cancer, renal, brain, gastric or preferably a colorectal, prostate,
breast, lung (especially non-small cell lung) or epidermoid, e.g.
epidermoid head and neck, especially mouth, cancer.
[0016] While the general treatment schedule allows for the
treatment of various tumor types already in front-line treatment,
the invention preferably relates to the treatment of tumors that
can be expected or have shown to be refractory to treatment with
other chemotherapeutics, e.g. standard treatment with one or more
other chemotherapeutics, especially with 5-fluorouracil and/or
taxane, e.g. TAXOL.RTM. treatment.
[0017] Surprisingly, it has now been found that even the
proliferation of tumor cells and tumors that are refractory to
standard treatment with other chemotherapeutics, e.g.
5-fluorouracil; and/or to treatment with a member of the taxane
class of compounds, most especially TAXOL.RTM., especially of a
colorectal tumor, especially one that is also refractory to
standard treatment, e.g. with 5-fluorouracil; or of a lung tumor,
especially a non-small cell lung cancer; an epidermoid, more
preferably epidermoid head and neck, such as mouth, tumor; or a
breast tumor; and/or metastasis thereof can be diminished or
stopped and that even regression or tumor disappearance is
possible.
DETAILED DESCRIPTION OF THE PREFERRED ASPECTS OF THE INVENTION
[0018] The present invention deals preferably with the following
subject matter as part of the invention:
[0019] Whenever within this whole specification "treatment of a
proliferative disease" or of a tumor, cancer or the like is
mentioned, there is meant
[0020] a) a method of treatment (=for treating) of a proliferative
disease, said method comprising the step of administering (for at
least one treatment) an epothilone, especially epothilone A and/or
B, especially B, (preferably in a pharmaceutically acceptable
carrier material) to a warm-blooded animal, especially a human, in
need of such treatment, in a dose that allows for the treatment of
said disease (=a therapeutically effective amount), preferably in a
dose (amount) as specified to be preferred hereinabove and
hereinbelow;
[0021] b) the use of an epothilone, preferably epothilone A and/or
B, especially epothilone B, for the treatment of a proliferative
disease; or an epothilone, especially epothilone B, for use in said
treatment (especially in a human);
[0022] c) the use of an epothilone, especially epothilone A and/or
B, especially epothilone B, for the manufacture of a pharmaceutical
preparation for the treatment of a proliferative disease;
and/or
[0023] d) a pharmaceutical preparation comprising a dose of an
epothilone, especially epothilone A and/or B, most especially
epothilone B, that is appropriate for the treatment of a
proliferative disease; or any combination of a), b), c) and d), in
accordance with the subject matter allowable for patenting in a
country where this application is filed;
[0024] e) a method of using an epothilone for the manufacture of a
pharmaceutical preparation for the treatment of a proliferative
disease, comprising admixing said epothilone with a
pharmaceutically acceptable carrier. In cases where a tumor disease
or a specific tumor (e.g. colon tumor, colon carcinoma or colon
cancer; or prostate tumor, prostate carcinoma or prostate cancer)
are mentioned instead of "proliferative disease", categories a) to
e) are also encompassed, meaning that the respective tumor disease
can be filled in under a) to e) above instead of "proliferative
disease", in accordance with the patentable subject matter.
[0025] In a first aspect, the present invention relates to the
treatment of a proliferative disease that is refractory to
treatment with one or more other chemotherapeutics, where an
epothilone, especially epothilone A and/or B, especially epothilone
B, is administered to a warm-blooded animal, especially a human,
preferably a human in need of such treatment, especially in a
therapeutically effective amount.
[0026] In a second aspect, the present invention relates to an in
vivo regimen for the treatment of a proliferative disease,
especially a cancer that is refractory to treatment with one or
more other chemotherapeutics, especially of the taxane class, like
TAXOL.RTM., and/or 5-fluorouracil, where an epothilone, especially
epothilone A and/or B, especially epothilone B, is administered in
a dose that is between about 1 and about 100%, preferably between
about 25 and 100%, of the (single administration) maximal tolerated
dose (MTD) to a warm-blooded animal, especially a human; and one or
more (preferably two to seven) further doses preferably each within
the dose range mentioned just above are administered in one or
preferably more than one further treatment cycle(s), especially
with an interval between the treatment cycles of one week or more
than one week after the preceding treatment, more preferably after
about one to about 6 weeks, most preferably about one to about
three weeks after the preceding treatment, respectively. Generally,
this treatment regimen where a high dose is administered in two or
more treatment cycles with periods of time between one to six,
preferably one to three weeks of time between administrations is
preferred over more frequent treatments with lower doses,
especially as it should reduce the frequency and duration of
hospitalization and as it shows superior antitumor effects and less
toxicity than more frequent treatments and more antitumor efficacy
than less frequent treatment.
[0027] Preferably, for epothilone B the dose in humans to be used
is calculated according to the formula (I)
single dose(mg/m.sup.2)=(0.1 to y).times.N (I)
[0028] where N (a whole (like 1, 2 or 3) or fractional number (like
1.5 or 2.3) wherever mentioned hereinbefore and hereinafter)) is
the number of weeks between treatments (preferably a number in the
range of about 1 to about 6 (corresponding to an interval of about
1 to about 6 weeks), especially about 1 to about 3 (corresponding
to a preferred interval of about 1 to about 3 weeks) and y is 6 or
preferably 5, more preferably 4.
[0029] More preferably, the treatment dose is calculated according
to the formula II
single dose(mg/m.sup.2)=(0.1 to 2.5).times.N; (II);
[0030] even more preferably according to the formula III,
single dose(mg/m.sup.2)=(0.1 to 1.7).times.N; (III);
[0031] or most preferably according to the formula IV
single dose(mg/m.sup.2)=(0.1 to 1).times.N (IV)
[0032] where, in each of formulae II to IV, N has the meaning given
under formula I. For the dosages calculated according to any of the
formulae I to IV, the following proviso must be met: The dose, even
if calculated higher, shall not exceed about 18 mg/m.sup.2 for a
single administration.
[0033] Preferably, for weekly treatment the dose is between about
0.1 and about 6, preferably about 0.1 and about 5 mg/M.sup.2, more
preferably about 0.1 and about 3 mg/M.sup.2, even more preferably
0.1 and 1.7 mg/M.sup.2, most preferably about 0.3 and about 1
mg/M.sup.2; for three-weekly treatment (treatment every three weeks
or every third week) the dose is between about 0.3 and about 18
mg/M.sup.2, preferably about 0.3 and about 15 mg/m.sup.2, more
preferably about 0.3 and about 12 mg/M.sup.2, even more preferably
about 0.3 and about 7.5 mg/m.sup.2, still more preferably about 0.3
and about 5 mg/m.sup.2, most preferably about 1.0 and about 3.0
mg/m.sup.2. This dose is preferably administered to the human by
intravenous (i.v.) administration during 2 to 180 min, preferably 2
to 120 min, more preferably during about 5 to about 30 min, most
preferably during about 10 to about 30 min, e.g. during about 30
min.
[0034] Preferably, especially in the case of weekly treatment, rest
periods of more than one week, more preferably of two to ten weeks,
more preferably three to six weeks after the preceding treatment
may be necessary after for example 3, 4, 6, 8, or more treatment
cycles, depending on patient condition, to allow for sufficient
recovery from the prec ding treatment.
[0035] In a third aspect of the invention, the present invention
relates to an in vivo regimen for the treatment of a proliferative
disease, especially one that is refractory to the treatment with
one or more other chemotherapeutics, where an epothilone,
preferably epothilone A and/or B, especially epothilone B, is
administered weekly to a warm-blooded animal, especially a human,
in a dose that is below 80%, more preferably below 50% of the
maximal tolerable dose (MTD).
[0036] In a fourth aspect, the invention relates to the in vivo
treatment of a proliferative disease that is refractory to the
treatment with one or more other chemotherapeutics, especially
5-fluorouracil or a microtubule stabilizing agent of the taxane
class, especially TAXOL.RTM., for example a multidrug resistant
tumor, where an epothilone, especially epothilone B, is
administered to a warm-blooded animal, especially a human.
[0037] In a fifth aspect, the invention relates to the in vivo
treatment of a proliferative disease, especially one that is
refractory to the treatment with one or more other
chemotherapeutics, by combined administration (a) of an epothilone,
preferably epothilone A and/or epothilone B, especially epothilone
B, in combination with (b) another antitumor chemotherapeutic,
preferably the combined treatment being timed so that component (a)
and (b) are administered to a warm-blooded animal, especially a
human (especially in need of such treatment), in combination in a
quantity which is jointly therapeutically effective against a
proliferative disease that preferably can be treated by
administration of an epothilone, more preferably epothilone A
and/or epothilone B, especially epothilone B; said administration
preferably taking place to a human that suffers from a tumor that
is refractory to other chemotherapeutic treatment, e.g. treatment
especially with 5-fluorouracil or especially with a member of the
taxane class of anti-cancer agents, like TAXOL.RTM..
[0038] In this regard, the invention also relates to a combination
preparation comprising components (a) and (b) as defined in the
last paragraph.
[0039] The invention also relates to a product which comprises
component (a) and component (b) as defined in the second paragraph
above, in the presence or absence of one or more pharmaceutically
acceptable carrier materials, as a combination preparation for
simultaneous or chronologically staggered administration to a
warm-blooded animal, especially a human, within a period of time
which is small enough for the active compounds both of component
(a) and of component (b) to mutually enhance antiproliferative
activity (especially against proliferating cells) in said
warm-blooded animal, for treating a proliferative disease.
[0040] The general terms used hereinbefore and hereinafter
preferably have the following meanings, if not defined
otherwise:
[0041] A proliferative disease is mainly a tumor disease (or
cancer) (and/or any metastases), wherever the tumor or the
metastasis are located), more especially a tumor selected from the
group comprising breast cancer, genitourinary cancer, lung cancer,
gastrointestinal cancer, epidermoid cancer, melanoma, ovarian
cancer, pancreas cancer, neuroblastoma, head and neck cancer (this
term, wherever it is used, meaning a head and/or neck cancer,
meaning that not only a cancer of head and neck, but also of head
or neck is envisaged) or bladder cancer, or in a broader sense
renal, brain or gastric cancer; more preferably (i) a tumor
selected from a breast tumor; an epidermoid tumor, especially and
epidermoid head and neck, preferably mouth, tumor; and a lung
tumor, especially a non-small cell lung tumor; or from a
gastrointestinal tumor, especially a colorectal tumor; and a
genitourinary tumor, especially a prostate tumor (especially a
hormone-refractory prostate tumor); or (ii) (more preferably) a
proliferative disease that is refractory to the treatment with
other chemotherapeutics, especially a corresponding tumor (and/or
any metastasis), more especially a tumor selected from the group
comprising tumors that are refractory to a standard treatment with
(an)other chemotherapeutic(s), especially with 5-fluorouracil
and/or (preferably) a microtubule stabilizing agent of the taxane
class, most especially TAXOL.RTM., still more preferably a tumor
selected from gastrointestinal, e.g. colorectal (especially
refractory to standard, e.g. 5-fluorouracil, and/or TAXOL.RTM.
treatment); and genitourinary, e.g. prostatic tumors (and/or a
metastasis thereof, especially a metastasis thereof); most
preferably a gastrointestinal tumor, especially a colorectal
cancer; or (iii) a tumor that is refractory to treatment with other
chemotherapeutics due to multidrug resistance, especially
refractory to a member of the taxane class of microtubule
stabilizing agents, preferably TAXOL.RTM., most especially a
multidrug, especially TAXOL.RTM., resistant lung tumor (especially
a non-small cell lung tumor), a multidrug resistant breast tumor,
or a multidrug resistant epidermoid, preferably epidermoid head and
neck, most preferably mouth, tumor.
[0042] In a broader sense of the invention, a proliferative disease
may furthermore be selected from hyperproliferative conditions such
as hyperplasias, fibrosis (especially pulmonary, but also other
types of fibrosis, such as renal fibrosis), angiogenesis,
psoriasis, atherosclerosis and smooth muscle proliferation in the
blood vessels, such as stenosis or restenosis following
angioplasty.
[0043] Where hereinbefore and subsequently a tumor, a tumor
disease, a carcinoma or a cancer are mentioned, also metastasis in
the original organ or tissue and/or in any other location are
implied alternatively or in addition, whatever the location of the
tumor and/or metastasis is.
[0044] The word "refractory" means that the respective
proliferative disease (especially a tumor and/or any metastasis
thereof, upon treatment with a (meaning at least one)
chemotherapeutic other than an epothilone, shows no or only weak
antiproliferative response (no or only weak inhibition of tumor
growth) after the treatment with such an agent, that is, a tumor
that cannot be treated at all or only with unsatisfying results
with other (preferably standard) chemotherapeutics (preferably as
defined above, especially 5-fluorouracil (especially in the case of
colorectal, like colon, cancer), antiandrogens or preferably
mitoxantrone (especially in the case of prostate cancer), or
antiestrogens, like letrozole (especially in the case of breast
cancer); or especially a member of the taxane class of
chemotherapeutics, e.g. TAXOTERE.RTM. or TAXOL.RTM., in a
warm-blooded animal, especially a human; for example the tumor
growth is not stopped, only retarded slightly or no regression is
found. The present invention, where treatment of refractory tumors
and the like is mentioned, is to be understood to encompass not
only (a) tumor(s) where one or more chemotherapeutics have already
failed during treatment of a patient, but also (a) tumor(s) that
can be shown to be refractory by other means, e.g. biopsy and
culture in the presence of chemotherapeutics. Where a term like
,,refractory against TAXOL.RTM." is used hereinbefore and
hereinafter, this term, in addition to the finished product, is
also intended to mean paclitaxel, the active substance of
TAXOL.RTM.. ,,Refractory to hormone treatment" or ,,hormone
refractory", in the case of a tumor of the genitourinary tract,
especially a prostate tumor, means refractory to treatment with an
antiandrogen.
[0045] TAXOL preferably stands for the finished product that
comprises paclitaxel, but, in a broader sense, is also meant to
encompass paclitaxel itself of any other paclitaxel formulation
with one or more carrier material(s).
[0046] Preferably, the term refractory means that with standard
dose a reduction of tumor growth by less than 50% (that is a T/C%
value of equal to or more than 50%) is obtained when compared with
a control without chemotherapeutic, e.g. by in vivo or in vitro
measurements.
[0047] Multidrug resistant tumor disease is one where resistance to
one or more chemotherapeutics, including those of the taxane class,
especially TAXOL.RTM., or the anthracycline class, especially
ADRIAMYCIN.RTM., is found. The basis for this resistance is the
export via an energy (especially ATP)-dependent pump located on the
surface of cells of the respective tumor, especially of the
P-glycoprotein family, especially P-glycoprotein (P-gp) itself. In
the present invention, alternatively or in addition other
mechanisms may cause a tumor to be refractory to treatment with
chemotherapeutics other than an epothilone. For example,
alterations in the drug target (especially microtubules in the
present case), changes in the intracellular metabolism that may
inactivate the compound, or changes in the physiology of the cell
that would facilitate by-passing or overriding of the mechanism of
drug action may lead to such resistance.
[0048] By the term "other chemotherapeutic" or ,,standard
chemotherapeutic", there is meant especially any chemotherapeutic
other than an epothilone; preferably one as defined in the
introduction, especially 5-fluorouracil (especially in the case of
colorectal, like colon, cancer), an anti-androgen or mitoxantrone
(especially in the case of prostate cancer), or an antiestrogen,
like letrozole (especially in the case of breast cancer);
especially, the term refers to 5-fluorouracil or (more preferably)
to members of the taxane class of microtubule stabilizing agents,
such as preferably Taxotere.RTM. or more preferably TAXOL.RTM..
,,Standard treatment with other chemotherapeutics", ,,other
chemotherapeutic treatment" or ,,standard chemotherapy" is
referring to treatment with at least one such ,,other" or
,,standard therapeutic".
[0049] By the term epothilone, any epothilone or epothilone
derivative is meant. Preferably, the term ,,epothilone" means
epothilone A, epothilone B, any epothilone derivative disclosed in
WO 98/25929 (which is incorporated by reference), or any mixture
thereof; more preferably, it means epothilone A and/or epothilone
B, and most preferably it relates to epothilone B.
[0050] The administration in all cases mentioned above and below
may take place orally but, in view of better and better defined
bioavailability, more preferably is made parenterally, especially
intravenously, e.g. by infusion or injection. Where subsequently
"infusion" is used, this means preferably intravenous infusion,
which is the most preferred mode of administration. Subsequently,
the data for adults are the basis for illustration. However, it
goes without saying that the present invention also relates to the
treatment of proliferative diseases in pediatrics. The doses must
then be corrected in accordance with standard methods and the age,
condition and other characteristics of the patient.
[0051] The Maximal Tolerated Dose (MTD) is determined according to
standard procedures; preferably, in warm-blooded animals the MTD in
case of oral or intravenous administration is determined as the
Dose of a single administration where no death occurs and a loss of
body weight of less than 40, preferably less than 25, percent (%)
is found in the treated warm-blooded animal individual (this term
here mainly referring to an animal; for humans see below).
[0052] The MTD may vary depending on the population of the patients
which may be defined by tumor type, age range, gender, tumor stage
and the like. While in animals, the most preferable way of
determining the MTD can be analogous to that shown in the Examples
presented below, in humans the MTD may generally be determined by
starting with one single administration of a very low dose, e.g.
{fraction (1/10)}th of the LD.sub.10 (i.e., the dose that is lethal
to 10% of animals) in the most sensitive animal species in which
toxicology studies have been performed, e.g. for epothilones
(especially epothilone B) in the range from 0.1 to 25 mg/M.sup.2,
especially for epothilone B in the range from 0.1 to 2.5
mg/M.sup.2, most especially in the range of 0.1 to 0.33 mg/M.sup.2.
Dose escalation for the next dose level is 100%, unless grade 2
toxicity is seen according to the US National Cancer Institute
Revised Common Toxicity Criteria, in which case dose escalation
will be 67%. Dose escalation for subsequent dose levels is in the
range of 25% to 67%. For example, three patients are usually
treated at one dose level and observed for acute toxicity for one
course of treatment before any more patients are entered. If none
of the three patients experience DLT (dose-limiting toxicity), then
the next cohort of three patients is treated with the next higher
dose. If two or more of the three patients experience DLT, then
three more patients are treated at the next lower dose unless six
patients have already been treated at that dose. If one of three
patients treated at a dose experiences DLT, then three more
patients are treated at the same level. If the incidence of DLT
among those patients is one in six, then the next cohort is treated
at the next higher dose. In general, if two or more of the six
patients treated at a dose level experience DLT, then the MTD is
considered to have been exceeded, and three more patients are
treated at the next lower dose as described above. The MTD is
defined as the highest dose studied for which the incidence of DLT
was less than 33%. Usually dose escalation for subsequent courses
in the same patient--i.e. intrapatient dose escalation--is not
permitted. Alternatively, dose steps may be defined by a modified
Fibonacci series in which the increments of dose for succeeding
levels beyond the starting dose are 100%, 67%, 50% and 40%,
followed by 33% for all subsequent levels. Finally, the MTD may be
found by methods described in Simon, R., et al., J. Nat. Cancer
Inst. 89(15), 1997, p. 1138-1147.
[0053] The DLT generally includes (but is not limited to) any
drug-related death and most drug-related grade 3 and 4 toxicities,
including febrile neutropenia (see also US National Cancer
Institute Revised Common Toxicity Criteria). See especially the
examples. For a human, the preferred treatment doses are defined by
formula I, more preferably formula II, most preferably formula III
mentioned above (with the proviso that no single dose is higher
than 18 mg/m.sup.2. Preferably, for weekly treatment the dose is
between about 0.1 and about 6, preferably about 0.1 and about 5
mg/M.sup.2, more preferably about 0.1 and about 3 mg/m.sup.2, even
more preferably about 0.1 and about 1.7 mg/m.sup.2, most preferably
about 0.3 and about 1 mg/m.sup.2; for three-weekly treatment
(treatment every third week) the dose is between about 0.3 and
about 18 mg/m.sup.2, preferably about 0.3 and about 15 mg/M.sup.2,
more preferably about 0.3 and about 12 mg/m.sup.2, even more
preferably about 0.3 and about 7.5 mg/m.sup.2, still more
preferably about 0.3 and about 5 mg/M.sup.2, most preferably about
1.0 and about 3.0 mg/m.sup.2. This dose is preferably administered
to the human by intravenous (i.v.) administration during 2 to 180
min, preferably 2 to 120 min, preferably during about 5 to about 30
min, most preferably during about 10 to about 30 min, e.g. about 30
min.
[0054] From the use of animal data, the applicable doses in (adult)
humans may be roughly calculated as follows:
[0055] A dose of 1 mg/kg in mouse corresponds to a dose of 3
mg/m.sup.2 in a human.
[0056] By sufficient recovery from the preceding treatment, in
warm-blooded animals there is meant especially the regain of body
weight of the treated individual to the starting level found before
the first dosing, preferably to at least 95% of said weight. In a
human, the recovery from each preceding dose administration is
preferably defined as recovery from any grade 3 or 4 toxicity,
including e.g. achievement of a platelet count of at least
100,000/mm.sup.3 and a neutrophil count of at least 1,500
cells/mm.sup.3 whole blood.
[0057] Treatment can be repeated if no response is achieved after a
first treatment, until tumor progression is found or until other
reasons (e.g. the condition of the patient) require termination of
treatment. In a human, the treatment with about 25 to about 100% of
the MTD is preferably repeated every 1 to ten, especially 2 to ten
weeks; preferably every 1 to 10 weeks, or every 3 to 6 weeks, until
disease progression, unacceptable toxicity, 1 or preferably 2
cycles beyond determination of a complete response, or patient
withdrawal of consent for any reason is encountered.
[0058] Preferably, in the case of weekly treatment of a human with
epothilone, the dose is in the range of about 5 to about 60%,
preferably about 10 to about 60%, e.g. about 5 to about 35% of the
MTD, especially in the range of about 30 to about 35% of the MTD.
Preferably, for epothilone B the dose is in the range of about 5 to
about 60%, more preferably about 10 to about 60%, especially in the
range of about 10 to about 45%, most especially in the range of
about 30 to about 45% of the three-weekly MTD.
[0059] More preferably, treatment is stopped after the third to
eighth, especially after the third to fifth weekly administration
followed by a rest period of two to five, e.g. two weeks before
further treatment is resumed, either by single or again weekly or
twice weekly administration. Especially, in the case of weekly
epothilone B treatment is stopped after the third to eighth
administration followed by a rest period of two to four, e.g. two
weeks before treatment is resumed by weekly administration.
[0060] Administration of component (a), that is epothilones A
and/or B, especially B, takes place preferably as described above,
especially using one of the special treatment regimens mentioned
above.
[0061] Administration of component (b) preferably takes place
according to treatment schedules that are known to the person
skilled in the art.
[0062] In one preferred embodiment, component (b) is administered
before component (a), preferably in a treatment comprising one or
more administrations of component (b) before starting the treatment
with component (a), preferably such that treatment with component
(b) ends at least two, preferably 5 to 10, e.g. about 5, days prior
to treatment with component (a) that is administered one or more
times thereafter, preferably one to five, especially one or two
times.
[0063] In a more preferred embodiment, component (a) is
administered on a 3-weekly schedule before component (b),
preferably in a treatment comprising one administration of
component (a) before starting the treatment with component (b),
more preferably such that treatment with component (a) ends
immediately prior to treatment with component (b) that is
administered thereafter.
[0064] In a second more preferred embodiment, component (a) is
administered on a weekly schedule. Component (b), on the other
hand, is administered on a 3-weekly schedule, with each
administration proceeding immediately upon completion of every
third administration of component (a).
[0065] In a third more preferred embodiment, component (a) is
administered on a weekly schedule before component (b), preferably
in a treatment comprising one administration of component (a)
before starting treatment with component (b), more preferably such
that treatment with component (a) ends immediately prior to
treatment with component (b) that is administered thereafter.
[0066] By the term "other chemotherapeutic agent" there is meant
especially any chemotherapeutic agent that is or can be used in the
treatment of tumor diseases, such as chemotherapeutics derived from
the following classes:
[0067] (A) Alkylating agents, preferably cross-linking
chemotherapeutics, preferably bis-alkylating agents,
[0068] (B) antitumor antibiotics, preferably doxorubicin
(ADRIAMYCIN.RTM., RUBEX.RTM.);
[0069] (C) antimetabolites;
[0070] (D) plant alkaloids;
[0071] (E) hormonal agents and antagonists,
[0072] (F) biological response modifiers, preferably lymphokines or
interferons
[0073] (G) inhibitors of protein tyrosine kinases and/or
serine/threonine kinases;
[0074] (H) antisense oligonucleotides or oligonucleotide
derivatives; or
[0075] (I) miscellaneous agents or agents with other or unknown
mechanism of action.
[0076] By the term "jointly therapeutically effective against a
proliferative disease that can be treated by administration of
epothilone A and/or epothilone B, especially epothilone B", there
is preferably meant a proliferative disease as mentioned above,
especially a tumor disease, the response preferably manifesting
itself in a diminished proliferation, e.g. diminished tumor growth
or even (more preferably) tumor regression or (most preferably)
tumor disappearance ("complete response").
[0077] Preferably, the term "quantity which is jointly
therapeutically effective against a proliferative disease that can
be treated by administration of epothilone A and/or epothilone B,
especially epothilone B" means any quantity of the components (a)
and (b) of the combinations that, in the combination, is
diminishing proliferation of cells responsible for any of the
mentioned proliferative diseases, especially tumor (including
metastatic) cells (especially diminished tumor growth) or,
preferably, even causing regression, more preferably even the
partial or complete disappearance, of such cells (especially tumor
regression, preferably complete response meaning disappearance of
the tumor(s)). This term not only comprises combinations of any
component (a) and (b) where (a) and (b) are dosed in such a manner
as to be antiproliferatively effective already without combination,
but also doses of any such component which alone would show no or
only marginal effect but which in combination leads to clearly
antiproliferative effects, that is to diminished proliferation or
preferably even to regression of the proliferating cells or even to
cure from the proliferative disease. In addition, here the term
"combination" is not only used to describe fixed combinations of
the components, but also any combination of components (a) and (b)
for simultaneous or chronologically staggered use within a period
of time which is small enough for the active compounds both of
component (a) and of component (b) to mutually enhance
antiproliferative activity, e.g. in a patient.
[0078] By the term "combination, preparation comprising component
(a) and (b)" there is meant any combination, be it as kit of parts
or as a single combined combination, of component (a) and (b) in
the form of a pharmaceutical product, that is preferably where a
pharmaceutically acceptable carrier material is present. For the
preferred carrier materials, see below under "Pharmaceutical
Preparations".
[0079] By the term "a product which comprises component (a) and
component (b)", there is preferably meant a product that
comprises
[0080] (a) at least one compound selected from epothilone A and
(preferably) epothilone B and
[0081] (b) at least one other chemotherapeutic agent in the
presence or absence of one or more pharmaceutically acceptable
carrier materials, as a combination preparation, for simultaneous
or chronologically staggered use, preferably within a period of
time which is small enough for the active compounds both of
component
[0082] (a) and of component (b) to mutually enhance
antiproliferative activity against proliferating cells, especially
in a patient, for treating a proliferative disease which responds
to such active compounds", especially a "kit of parts" in the sense
that the effective components (a) and (b) of the combination can be
dosed independently or by use of different fixed combinations with
distinguished amounts of any components (a) and (b) at different
time points. The "parts" of the kit of parts can then be
administered simultaneously or chronologically staggered, that is
at different time points and with equal or different time intervals
for any part of the kit of parts, preferably with the condition
that the time intervals are chosen such that the effect on the
proliferative disease in the combined use of the parts is larger
than the effect which would be obtained by use of only any one of
component (a) and (b) alone or by use of both in a way that the
compounds act independently (e.g. with long enough periods to avoid
effects of each of the components on the others), that is, stronger
inhibition of proliferation or, preferably, stronger regression or
even cure of the proliferative disease is found than when the same
dose of only one of components (a) and (b) is administered alone in
the same dose or after sufficiently long time intervals that mutual
effects of the components (a) and (b) are excluded. That is meant
by the term "to mutually enhance antiproliferative activity against
proliferating cells, especially in a patient"; preferably, there is
meant a mutual enhancement of the effect of the components (a) and
(b), especially a synergism and/or the causing of regression of the
proliferating cells, up to and including their complete
destruction, and especially a strong synergism between components
(a) and (b).
[0083] By the term "proliferating cells", especially pathologically
or abnormally proliferating cells are meant, such as tumor and/or
tumor metastasis cells, especially of tumors as defined herein as
being preferred.
[0084] Preferred are combinations which show enhanced
antiproliferative activity when compared with the single components
alone, especially combinations that show synergism (synergistic
combinations) or combinations that lead to regression of
proliferative tissues and/or cure from proliferative diseases.
[0085] The term "synergism" is standing for an effect that is
stronger than additive, that is, a stronger effect of the
combination of any component (a) with any component (b) than could
be reached by the factor of diminuation of proliferation obtained
from mere multiplication of the factor of diminuation of
proliferation for any component (a) alone or any component (b)
alone when compared to a control without treatment when each (a)
and (b) as such, whether alone or in combination, is administered
in the same dose as in the single treatment without combination
(which does not mean that the dose of (a) must be identical to that
of (b), although this may also be the case). As theoretical example
for mere illustration, if a component (a) alone gives a growth of
tumor cells that is diminished by a factor of 2 in comparison to a
control without any treatment and a component (b) alone gives a
diminuation of growth by a factor of 1.5, then an additive effect
would be one where, by combined use of component (a) and component
(b), a 3-fold diminuation of growth would be found (multiplication
of 2 with 1.5). A synergistic effect would for example be present
if a more than 3-fold diminuation of proliferation is found. The
presence of synergism can be shown by this fractional product
method [Webb, in: ,,Enzymes and Metabolic Inhibitors", Vol. 1,
66-73 and 488-512, Academic Press, New York] or alternatively by
the isobologram method [see references in: Berenbaum Pharmacol.
Rev. 41, 99-141 (1984)], and/or the combination index (CI)
calculation method [Chou et al., Trends Pharmacol. Sci. 4, 450-454
(1983); or Chou et al., New Avenues in Developmental Cancer
Chemotherapy; Bristol-Myers Symposium Series, K. R. Harrap and I.
A. Connors (eds.), 37-64, New York, Academic Press (1987)].
[0086] The term "pharmaceutically acceptable carrier materials" is
explained below in the definition of pharmaceutical
preparations.
[0087] Provided that in the respective molecule salt-forming groups
are present, component (b) (other chemotherapeutic(s)) may also be
present in the form of salts wherever it is mentioned above or
below.
[0088] Termination of treatment preferably takes place when either
of the following occurs: Disease progression, for example under the
Southwest Oncology Group (SWOG) response criteria; unacceptable
toxicity (e.g. to the patient, the investigator, or both);
treatment 2 cycles beyond determination of a complete response, for
example under the Southwest Oncology Group (SWOG) response
criteria; or patient withdrawal of consent.
[0089] Salts of components are especially acid addition salts,
salts with bases or, when several salt-forming groups are present,
optionally also mixed salts or internal salts. Salts are especially
the pharmaceutically acceptable, e.g. substantially non-toxic,
salts.
[0090] Such salts are formed, for example, from chemotherapeutics
having an acidic group, for example a carboxy, phosphodiester or
phosphorothioate group, and are, for example, their salts with
suitable bases, such as non-toxic metal salts derived from metals
of groups Ia, Ib, IIa and IIb of the Periodic Table of Elements,
especially suitable alkali metal salts, for example lithium, sodium
or potassium salts, or ammonium salts, also those salts that are
formed with organic amines, such as unsubstituted or
hydroxy-substituted mono-, di- or tri-alkylamines, especially
mono-, di- or tri-lower alkylamines, or with quaternary ammonium
compounds, for example with N-methyl-N-ethylamine, diethylamine,
triethylamine, mono-, bis- or tris-(2-hydroxy-lower alkyl)amines,
such as mono-, bis- or tris-(2-hydroxyethyl)amine,
2-hydroxy-tert-butylamine or tris(hydroxymethyl)methylamine,
N,N-di-lower alkyl-N-(hydroxy-lower alkyl)-amines, such as
N,N-dimethyl-N-(2-hydroxyet- hyl)-amine or
tri-(2-hydroxyethyl)amine, or N-methyl-D-glucamine, or quaternary
ammonium salts, such as tetrabutylammonium salts. The
chemotherapeutics having a basic group, for example an amino or
imino group, can form acid addition salts, for example with
inorganic acids, for example a hydrohalic acid, such as
hydrochloric acid, sulfuric acid or phosphoric acid, or with
organic carboxylic, sulfonic, sulfo or phospho acids or
N-substituted sulfamic acids, such as, for example, acetic acid,
propionic acid, glycolic acid, succinic acid, maleic acid,
hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid,
tartaric acid, gluconic acid, citric acid, or benzoic acid, also
with amino acids, for example, .alpha.-amino acids, and also with
methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic
acid, ethane-1,2-disulfonic acid, benzenesulfonic acid,
4-methylbenzenesulfonic acid, naphthalene-2-sulfonic acid,
N-cyclohexylsulfamic acid (with formation of the cyclamates) or
with other acidic organic compounds, such as ascorbic acid.
Compounds having acidic and basic groups can also form internal
salts. If more than one salt-forming group is present, it is also
possible that mixed salts are present.
[0091] Where hereinbefore and hereinafter numerical terms are used,
they are meant to include the numbers representing the upper and
lower limits. For example, ,,between 1 and 3" stands for a range
,,from and including 1 up to and including 3", and ,,in the range
from 1 to 3" would stand for ,,from and including 1 up to and
including 3". The same is true where instead of numbers (e.g. 3)
words denoting numbers are used (e.g. ,,three").
[0092] Where ,,comprising" is used, this can preferably be replaced
by ,,consisting essentially of", more preferably by ,,consisting
of".
[0093] Where ,,about" is used in connection with a number, this
preferably means the number .+-.15%, more preferably the number
plus 5%, most preferably the number itself without ,,about". For
example, ,,about 100" would stand for ,,from and including 85 to
and including 115". Where ,,about" is used in connection with
numeric ranges, for example ,,about 1 to about 3", or ,,between
about one and about three", preferably the definition of about"
given for a number in the last sentence is applied to each number
defining the start and the end of a range separately. Preferably,
where ,,about" is used in connection with any numerical values, the
,,about" can be deleted.
[0094] ,,Weekly" stands for ,,about once a week" (meaning that more
than one treatment is made with an interval of about one week
between treatments), the about here preferably meaning.+-.1 day
(that is, translating into ,,every 6 to 8 days"); most preferably,
"weekly" stands for ,,once every 7 days".
[0095] ,,3-weekly" or ,,three-weekly" stands for ,,about once every
three weeks" (meaning that more than one treatment is made with an
interval of about three weeks between treatments), the about here
preferably meaning.+-.3 days (that is, translating into every 18 to
24 days); most preferably, ,,weekly" stands for ,,once every 21
days" (=every third week).
PREFERRED EMBODIMENTS OF THE INVENTION
[0096] In the following preferred embodiments of the invention, the
general definitions may be replaced by the more specific
definitions given hereinbefore and hereinafter, as appropriate.
[0097] (1) The present invention relates especially to the
treatment of a proliferative disease, especially a cancer,
especially a cancer that is refractory to treatment with other
chemotherapeutics and/or a member of the taxane class of
anti-cancer agents, especially TAXOL.RTM., more especially one of
the preferred diseases as defined above or below, characterized in
that an epothilone, especially epothilone A or most especially
epothilone B, is administered more than once with a weekly up to
three-weekly interval to a human in a dose that is calculated
according to the formula (I)
single dose(mg/m.sup.2)=(0.1 to y).times.N (I)
[0098] where N (a whole or fractional number) is the number of
weeks between treatments (about one to about three weeks), that is
N is about 1 to about 3; more preferably, the treatment dose is
calculated according to the formula II
single dose(mg/m.sup.2)=(0.1 to 2.5).times.N; (II);
[0099] even more preferably according to the formula III,
single dose(mg/m.sup.2)=(0.1 to 1.7).times.N; (III);
[0100] or still more preferably according to the formula IV
single dose(mg/m.sup.2)=(0.1 to 1).times.N (IV)
[0101] where, in each of formulae II to IV, N is about 1 to about 3
(corresponding to intervals of about 1 to about 3 weeks between
treatments);
[0102] the epothilone, especially epothilone B, administration
preferably taking place
[0103] (a) weekly in a human in a dose that lies between about 0.1
and about 6, preferably about 0.1 and about 5 mg/M.sup.2, more
preferably about 0.1 and about 3 mg/M.sup.2, even more preferably
about 0.1 and about 1.7 mg/M.sup.2, most preferably about 0.3 and
about 1 mg/m.sup.2; or
[0104] (b) three-weekly in a human in a dose that lies dose is
between about 0.3 and about 18 mg/m.sup.2, preferably about 0.3 and
about 15 mg/M.sup.2, more preferably about 0.3 and about 12 mg/M2,
even more preferably about 0.3 and about 7.5 mg/M2, still more
preferably about 0.3 and about 5 mg/M2, most preferably about 1.0
and about 3.0 mg/M.sup.2;
[0105] the administration preferably taking place by i.v. infusion
during 2 to 120 min, more preferably during about 5 to about 30
min, most preferably during about 10 to about 30 min, e.g. during
about 30 min.
[0106] (2) The present invention preferably relates also to the
treatment of a tumor disease that is refractory to the treatment
with an other chemotherapeutic, especially selected from
5-fluorouracil and preferably a microtubule stabilizing agent of
the taxane class, most especially TAXOL.RTM., said tumor being
selected from a gastrointestinal, e.g. colorectal; a renal; a
genitourinary, e.g. prostatic; a pancreatic; and a brain tumor
(and/or any metastasis thereof), most preferably a gastrointestinal
tumor, especially a colorectal cancer, more especially a
gastrointestinal cancer, especially a colorectal cancer, that is
refractory to treatment with a member of the taxane class of
anti-cancer agents, especially TAXOL.RTM., or very especially such
tumor that is refractory to a standard chemotherapy, such as
treatment a standard chemotherapeutic, especially with
5-fluorouracil; or a tumor of the genitourinary tract, especially a
prostate cancer, most especially a hormone-refractory prostate
cancer; where epothilone A and/or B, especially epothilone B, is
administered to a warm-blooded animal, especially a human.
[0107] (3) The present invention preferably also relates to the
treatment of a tumor disease, especially a lung tumor, especially a
non-small cell lung carcinoma, especially such lung cancer that is
refractory to treatment with a member of the taxane class of
anti-cancer agents, especially TAXOL.RTM.; a breast tumor,
especially one that is multidrug resistant; or an epidermoid tumor,
preferably an epidermoid head and neck, especially mouth, tumor,
especially if the latter is multidrug resistant and/or resistant to
treatment with a member of the taxane class of anti-cancer agents,
in particular TAXOL.RTM.; where epothilone A and/or B, especially
epothilone B, is administered to a warm-blooded animal, especially
a human.
[0108] (4) The present invention also preferably relates to an in
vivo regimen for the treatment of a tumor disease, especially (i)
of a tumor of the gastrointestinal tract, most especially a tumor
of the colon and/or rectum (colorectal tumor); and/or (ii) a tumor
of the genitourinary tract, especially a prostate tumor (preferably
a hormone-refractory prostate tumor); especially where such tumor
is refractory to treatment with an other chemotherapeutic,
especially one of the taxane class, most especially TAXOL.RTM.;
where epothilone A and/or B, especially epothilone B, is
administered once in a dose that is between about 20 and about 100%
of the MTD, to a human; and, if required, one or more (preferably
two to seven) further doses each within the dose range mentioned
above for the first dose are administered in further treatment
cycles, preferably each dose after a period of time that allows for
sufficient recovery of the treated individual from each preceding
dose administration, especially more than two weeks after the
preceding treatment, more especially two to 10 weeks, most
especially three to six weeks after the preceding treatment,
especially three weeks after that treatment.
[0109] More preferably, under (1) to (4) epothilone B is
administered weekly to a human in a dose that lies between about
0.1 and about 6, preferably about 0.1 and about 5 mg/M.sup.2, more
preferably about 0.1 and about 3 mg/M.sup.2, even more preferably
about 0.1 and about 1.7 mg/m.sup.2, most preferably about 0.3 and
about 1 mg/m.sup.2; or epothilone B is administered three-weekly
(every 3 weeks) in a dose that is between about 0.3 and about 18
mg/m.sup.2, preferably about 0.3 and about 15 mg/M.sup.2, more
preferably about 0.3 and about 12 mg/m.sup.2, even more preferably
about 0.3 and about 7.5 mg/M.sup.2, still more preferably about 0.3
and about 5 mg/m.sup.2, most preferably about 1.0 and about 3.0
mg/M.sup.2. This dose is preferably administered to the human by
intravenous (i.v.) administration during 2 to 120 min, more
preferably during about 5 to about 30 min, most preferably during
about 10 to about 30 min, e.g. during about 30 min.
[0110] More preferably, said treatment is repeated every about 1 to
about 3 weeks, until disease progression, unacceptable toxicity, 1
or preferably 2 cycles beyond determination of a complete response,
or patient withdrawal of consent for any reason is encountered.
[0111] (5) The present invention preferably also relates to an in
vivo regimen for the treatment of a tumor disease, especially (i)
of a tumor of the gastrointestinal tract, most especially a tumor
of the colon and/or rectum (colorectal tumor); and/or (ii) a tumor
of the genitourinary tract, especially a prostate tumor; especially
where such tumor is refractory to treatment with an other
chemotherapeutic, especially one of the taxane class, most
especially TAXOL.RTM. (preferably a hormone-refractory prostate
tumor); where epothilone A and/or B, especially epothilone B, is
administered weekly to a warm-blooded animal in a dose that is
below 80%, more preferably below 50% of the maximal tolerable dose
(MTD).
[0112] Preferably, in the case of weekly treatment of a human with
said epothilone(s), the dose is in the range of about 1 to about
60%, preferably about 10 to about 60%, e.g. about 5 to about 35% of
the MTD, for example in the range of about 30 to about 35% of the
MTD. Preferably, for epothilone B the dose is in the range of about
5 to about 60%, preferably about 10 to about 60%, especially in the
range of about 10 to about 45%, most especially in the range of
about 30 to about 45% of the three-weekly MTD. of the three-weekly
MTD. In a special case, the dose can be between about 2 and about
18 mg/m.sup.2 for epothilone B.
[0113] More preferably, treatment is stopped after the third to
eighth, especially after the third to fifth weekly administration
followed by a rest period of two to five, e.g. two weeks before
further treatment is resumed. Preferably and if required, in the
case of weekly epothilone B administration treatment is stopped
after the third to eighth administration followed by a rest period
of two to four, e.g. two weeks before treatment is resumed by
weekly administration.
[0114] (6) The invention preferably also relates to the in vivo
treatment of a tumor disease by combined administration (a) of
epothilone A and/or epothilone B, especially epothilone B, in
combination with (b) an other chemotherapeutic agent selected from
the group consisting of (A) alkylating agents, preferably
cross-linking chemotherapeutics, preferably bis-alkylating
agents,
[0115] (B) antitumor antibiotics, preferably doxorubicin
(ADRIAMYCIN.RTM., RUBEX.RTM.);
[0116] (C) antimetabolites;
[0117] (D) plant alkaloids;
[0118] (E) hormonal agents and antagonists,
[0119] (F) biological response modifiers, preferably lymphokines or
interferons
[0120] (G) inhibitors of protein tyrosine kinases and/or
serine/threonine kinases;
[0121] (H) antisense oligonucleotides or oligonucleotide
derivatives; or
[0122] (I) miscellaneous agents or agents with other or unknown
mechanism of action;
[0123] the combined treatment being timed so that component (a) and
(b) are combined for simultaneous or chronologically staggered use
within a period of time which is small enough for the active
compounds both of component (a) and of component (b) to mutually
enhance antiproliferative activity, e.g. in a patient.
[0124] (7) The invention also relates to a product which comprises
component (a) and component (b) as defined under (6) above, in the
presence or absence of one or more pharmaceutically acceptable
carrier materials, as a combination preparation for simultaneous or
chronologically staggered administration to a human within a period
of time which is small enough for the active compounds both of
component (a) and of component (b) to mutually enhance activity
against a tumor disease, especially (i) a tumor of the
gastrointestinal tract, most especially a tumor of the colon and/or
rectum (colorectal tumor); and/or (ii) a tumor of the genitourinary
tract, especially a prostate tumor; especially where such tumor is
refractory to treatment with an other chemotherapeutic, especially
one of the taxane class, most especially TAXOL.RTM.; for treating
said tumor disease.
[0125] Under (1) to (7) or the subsequent embodiments of the
invention, administration of the epothilone, especially epothilone
B, preferably takes place by infusion, especially by intravenous
infusion.
[0126] The following are some especially preferred embodiments of
the invention:
[0127] A1. The use of an epothilone, especially epothilone A and/or
epothilone B, for the treatment of a proliferative disease that is
refractory to treatment with other chemotherapeutics; or the use of
said epothilone, especially epothilone B, for the manufacture of a
pharmaceutical preparation for the treatment of a proliferative
disease that is refractory to the treatment with other
chemotherapeutics.
[0128] A2. The use according to A1 where the proliferative disease
is a tumor disease that is refractory to a microtubule stabilizing
agent of the taxane class, especially TAXOL.RTM..
[0129] A3. The use according to any one of A1 to A2 where the
proliferative disease is a colorectal tumor, and/or a metastasis
thereof.
[0130] A4. The use according to any one of A1 to A2 where the
proliferative disease is a prostatic tumor, and/or a metastasis
thereof; especially a hormone-refractory prostate tumor.
[0131] B1. The use of an epothilone for the manufacture of a
pharmaceutical preparation that is appropriate for administration
of said epothilone once in a dose that is between about 1 and about
100% of the maximal tolerated dose (MTD) in a warm-blooded animal,
to said warm-blooded animal for the treatment of a proliferative
disease that is refractory to the treatment with other
chemotherapeutics.
[0132] B2. The use according to B1 where the epothilone is
epothilone A and/or epothilone B, preferably epothilone B.
[0133] B3. The use according to any one of B1 and B2 where the dose
is between 25 and 100% of the maximal tolerated dose and the
warm-blooded animal is a human.
[0134] B4. The use according to any one of B1 to B3 where the dose
unit for an adult human is in the range of between about 0.3 and
about 18, preferably about 0.3 and about 12, more preferably about
0.3 and about 7.5, most preferably about 1.0 and about 3 mg/m.sup.2
of epothilone B with three-weekly treatment; or between about 0.1
and about 6, preferably about 0.1 to about 5, more preferably about
0.1 and about 3, most preferably about 0.3 and about 1 mg/m.sup.2
for weekly treatment.
[0135] B5. The use according to any one of B1 to B4 where the dose
is chosen so that after a period of time that allows for sufficient
recovery of the treated individual from each preceding dose a
further dose can be administered.
[0136] B6. The use according to any one of B1 to B5 where the
proliferative disease is a tumor.
[0137] B7. The use according to any one of B1 to B6 where the
proliferative disease is a colorectal tumor, and/or a metastasis
thereof.
[0138] B8. The use according to any one of B1 to B6 where the
proliferative disease is a prostate tumor, and/or a metastasis
thereof.
[0139] B9. The use according to any one of B1 to B8 where the tumor
is one that is refractory to treatment with a microtubule
stabilizing agent of the taxane, microtubule stabilizing class of
agents, especially TAXOL.RTM..
[0140] C1. The use of an epothilone, preferably epothilone A and/or
epothilone B, especially epothilone B, for the manufacture of a
pharmaceutical preparation that is appropriate for administration
of said epothilone once weekly, where the dose is 80% or less,
preferably 50% or less, of the MTD.
[0141] D1. The use of an epothilone, especially epothilone A and/or
epothilone B, for the manufacture of a pharmaceutical preparation
that is appropriate for the combined administration (a) of an
epothilone, preferably epothilone A and/or epothilone B, in
combination with (b) another antitumor chemotherapeutic to a
warm-blooded animal that suffers from a proliferative disease that
is refractory to the treatment with other chemotherapeutics,
especially a colorectal or prostate tumor and/or a metastasis
thereof.
[0142] E1. A pharmaceutical preparation for the treatment of a
proliferative disease, especially a tumor disease, especially one
of those characterized as being preferred above or below, in a
human, said preparation comprising an epothilone, especially
epothilone B, in a dose ranging from 1 to 100%, preferably from 25
to 100% of the maximal tolerable dose (MTD), and a pharmaceutically
acceptable carrier.
[0143] F1. A combination preparation comprising (a) epothilone A or
preferably epothilone B and (b) one or more other antitumor
chemotherapeutics, and a pharmaceutically acceptable carrier.
[0144] G1. A product which comprises as component (a) epothilone A
and/or B, preferably epothilone B, and as component (b) any other
antitumor chemotherapeutic, in the presence or absence of one or
more pharmaceutically acceptable carrier materials, as a
combination preparation for simultaneous or chronologically
staggered administration to a warm-blooded animal, especially a
human, within a period of time which is small enough for the active
compounds both of component (a) and of component (b) to mutually
enhance antitumor activity in said warm-blooded animal, for
treating a proliferative disease.
[0145] The invention relates most especially to the treatment of
following tumor/cancer types with epothilone B:
[0146] (i) a gastrointestinal, especially a colorectal tumor that
is refractory to a representative of the taxane class of
anti-cancer agents, in particular TAXOL.RTM.; or more especially to
the treatment with standard chemotherapy, especially with
5-fluorouracil, and/or TAXOL.RTM..
[0147] (ii) a tumor of the genitourinary tract, especially a
prostate tumor, including primary and especially metastatic tumors;
more especially if refractory to hormone treatment;
[0148] (iii) an epidermoid, more especially epidermoid head and
neck, most especially epidermoid mouth tumor, especially one of
these that is refractory to treatment with an other
chemotherapeutic, especially due to multi-drug resistance,
especially to treatment with a member of the taxane class of
anti-cancer agents, especially TAXOL.RTM.;
[0149] (iv) a lung tumor, especially a non-small cell lung cancer,
that is refractory to treatment with an other chemotherapeutic,
especially due to (mainly) multi-drug resistance, especially to
treatment with a member of the taxane class of anti-cancer agents,
especially TAXOL.RTM.; and/or
[0150] (v) a breast tumor, especially a breast tumor that is
multidrug resistant, more especially one that is refractory to
treatment with a member of the taxane class of anti-cancer agents,
especially TAXOL.RTM..
[0151] Preferably, the invention relates to the treatment of any
one of the above-mentioned tumor types (i) to (v), most preferably
to that of (i), (ii), (iv) and (v).
[0152] More preferably, the invention relates to the treatment of
any of the tumor types mentioned above under (i) to (v), especially
to any one of them, by treatment with an intravenous infusion of
epothilone B over 2 to 120 min, preferably during about 5 to about
30 min, more preferably during about 10 to about 30 min, most
preferably during about 30 min;
[0153] said administration being repeated every one to three weeks,
preferably every one week (weekly) or every three weeks;
[0154] where the epothilone B dose preferably is defined by formula
I,
single dose(mg/m.sup.2)=(0.1 to y).times.N (I)
[0155] where N (a whole or fractional number) is the number of
weeks between treatments that lies between about one and about
three weeks, that is, N is about 1 to about 3; and y is 6,
preferably 5, more preferably 4.
[0156] More preferably, the treatment dose is calculated according
to the formula II
single dose(mg/m.sup.2)=(0.1 to 2.5).times.N; (II);
[0157] even more preferably according to the formula III,
single dose(mg/m.sup.2)(0.1 to 1.7).times.N; (III);
[0158] or most preferably according to the formula IV
single dose(mg/m.sup.2)=(0.1 to 1).times.N (IV)
[0159] where, in each of formulae 11 to IV, N corresponds to about
1 to about 3 (standing preferably for weekly up to three-weekly
treatment).
[0160] More preferably, for weekly treatment the dose is between
about 0.1 and about 6, preferably about 0.1 to about 5 mg/m.sup.2,
more preferably about 0.1 and about 3 mg/m.sup.2, even more
preferably about 0.1 and about 1.7 mg/m.sup.2, most preferably
about 0.3 and about 1 mg/m.sup.2; or for three-weekly treatment
between about 0.3 and about 18 mg/m.sup.2, preferably about 0.3 and
about 15 mg/m.sup.2, more preferably about 0.3 and about 12
mg/m.sup.2, even more preferably about 0.3 and about 7.5
mg/M.sup.2, still more preferably about 0.3 and about 5 mg/M2, most
preferably about 1.0 and about 3.0 mg/M.sup.2.
[0161] More preferably, rest periods of more than one week, more
preferably of two to ten weeks, more preferably three to six weeks
after the preceding treatment may be necessary (especially in the
case of weekly treatment) after for example 3, 4, 6, 8, or more
cycles, depending on patient condition, to allow for sufficient
recovery from the preceding treatment.
[0162] Especially preferred are also treatment conditions and
formulations in analogy to those mentioned in the Examples.
[0163] Pharmaceutical Formulations
[0164] The present invention also relates to the use of epothilone
A and/or B, especially epothilone A or preferably epothilone B, for
the manufacture of a pharmaceutical formulation for use against a
tumor disease as defined above; or to a pharmaceutical formulation
for the treatment of said tumor disease comprising epothilone A
and/or B, especially epothilone A or preferably epothilone B, and a
pharmaceutically acceptable carrier.
[0165] The invention relates also to pharmaceutical compositions
comprising epothilone A and/or epothilone B, especially epothilone
B, for the treatment of a proliferative disease, especially a tumor
disease defined as being preferred above, and to the preparation of
pharmaceutical preparations for said treatment.
[0166] Epothilone A and/or B may be used, for example, for the
preparation of pharmaceutical compositions that comprise an
effective amount of the active ingredient together or in admixture
with a significant amount of inorganic or organic, solid or liquid,
pharmaceutically acceptable carriers.
[0167] The invention relates also to a pharmaceutical composition
that is suitable for administration to a warm-blooded animal,
especially a human, for the treatment of a proliferative disease as
defined hereinbefore, comprising an amount of epothilone A and/or
B, especially epothilone B, which is effective for the treatment of
said proliferative disease, together with at least one
pharmaceutically acceptable carrier.
[0168] The pharmaceutical compositions according to the invention
are those for enteral, such as nasal, rectal or oral, or preferably
parenteral, such as intramuscular or intravenous, administration to
a warm-blooded animal (human or animal), that comprise an effective
dose of the pharmacologically active ingredient, alone or together
with a significant amount of a pharmaceutically acceptable carrier.
The dose of the active ingredient depends on the species of
warm-blooded animal, the body weight, the age and the individual
condition, individual pharmacokinetic data, the disease to be
treated and the mode of administration; preferably, the dose is one
of the preferred doses as defined above, being accomodated
appropriately where pediatric treatment is intended.
[0169] The pharmaceutical compositions comprise from about 0.00002
to about 95%, especially (e.g. in the case of infusion dilutions
that are ready for use) of 0.0001 to 0.02%, or (for example in case
of infusion concentrates) from about 0.1% to about 95%, preferably
from about 20% to about 90%, active ingredient (weight by weight,
in each case). Pharmaceutical compositions according to the
invention may be, for example, in unit dose form, such as in the
form of ampoules, vials, suppositories, dragees, tablets or
capsules.
[0170] Preferably, the dose is chosen so as to allow for the
treatment regimen based on the MTD mentioned above for single or
rare treatment of a tumor disease.
[0171] The pharmaceutical compositions of the present invention are
prepared in a manner known per se, for example by means of
conventional dissolving, lyophilizing, mixing, granulating or
confectioning processes.
[0172] Solutions of the active ingredient, and also suspensions,
and especially isotonic aqueous solutions or suspensions, are
preferably used, it being possible, for example in the case of
lyophilized compositions that comprise the active ingredient alone
or together with a pharmaceutically acceptable carrier, for example
mannitol, for such solutions or suspensions to be produced prior to
use. The pharmaceutical compositions may be sterilized and/or may
comprise excipients, for example preservatives, stabilizers,
wetting and/or emulsifying agents, solubilizers, salts for
regulating the osmotic pressure and/or buffers, and are prepared in
a manner known per se, for example by means of conventional
dissolving or lyophilizing processes. The said solutions or
suspensions may comprise viscosity-increasing substances, such as
sodium carboxymethylcellulose, carboxymethylcellulose, dextran,
polyvinylpyrrolidone or gelatin.
[0173] Suspensions in oil comprise as the oil component the
vegetable, synthetic or semi-synthetic oils customary for injection
purposes. There may be mentioned as such especially liquid fatty
acid esters that contain as the acid component a long-chained fatty
acid having from 8 to 22, especially from 12 to 22, carbon atoms,
for example lauric acid, tridecylic acid, myristic acid,
pentadecylic acid, palmitic acid, margaric acid, stearic acid,
arachidic acid, behenic acid or corresponding unsaturated acids,
for example oleic acid, elaidic acid, erucic acid, brasidic acid or
linoleic acid, if desired with the addition of antioxidants, for
example vitamin E, .beta.-carotene or
3,5-di-tert-butyl-4-hydroxytoluene. The alcohol component of those
fatty acid esters has a maximum of 6 carbon atoms and is a mono- or
poly-hydroxy, for example a mono-, di- or tri-hydroxy, alcohol, for
example methanol, ethanol, propanol, butanol or pentanol or the
isomers thereof, but especially glycol and glycerol. The following
examples of fatty acid esters are therefore to be mentioned: ethyl
oleate, isopropyl myristate, isopropyl palmitate, "Labrafil M 2375"
(polyoxyethylene glycerol trioleate, Gattefoss, Paris), "Miglyol
812" (triglyceride of saturated fatty acids with a chain length of
C.sub.8 to C.sub.12, Huls AG, Germany), but especially vegetable
oils, such as cottonseed oil, almond oil, olive oil, castor oil,
sesame oil, soybean oil and more especially groundnut oil.
[0174] The injection or infusion compositions are prepared in
customary manner under sterile conditions; the same applies also to
introducing the compositions into ampoules or vials and sealing the
containers.
[0175] Preferred is an infusion formulation comprising epothilone A
and/or epothilone B, especially epothilone B, and a
pharmaceutically acceptable organic solvent.
[0176] The formulation does not require the use of a surfactant.
Surfactants such as Cremophor may cause allergic reactions and they
also can leach plasticizers from standard PVC containers, tubing
and the like. Consequently, when they are employed one is required
to use special infusion apparatus, e.g. nitro-glycerine tubing and
non-plastizised containers, such as glass, tubing and the like.
[0177] The pharmaceutically acceptable organic solvent used in a
formulation according to the invention may be chosen from any such
organic solvent known in the art. Preferably the solvent is
selected from alcohol, e.g. absolute ethanol or ethanol/water
mixtures, more preferably 70% ethanol, polyethylene glycol 300,
polyethylene glycol 400, polypropylene glycol or
N-methylpyrrolidone, most preferably polypropylene glycol or 70%
ethanol or especially polyethylene glycol 300.
[0178] The Epothilones may preferably be present in the formulation
in a concentration of about 0.1 to about 100 mg/ml, more preferably
about 1 to about 100 mg/ml, still more preferably about 1 to about
10 mg/ml (especially in infusion concentrates).
[0179] Epothilone A and Epothilone B may be used as pure substances
or as a mixture of Epothilone A and B. Given the greater
anti-tumour activity of Epothilone B it may be employed in a lower
concentration than Epothilone A in the formulation. When used in
its pure form it is preferable to employ a concentration of
Epothilone A of 5 to 100 mg/ml, preferably 10 to 50 mg/ml, whereas
when Epothilone B is used in its pure form it is preferably
employed in a concentration of 0.1 to 10, more preferably 1 to 10,
still more preferably 1 to 2 mg/ml (this number makes reference
especially to an infusion concentrate that, before treatment, is
diluted accordingly, see below).
[0180] Such formulations are conveniently stored in vials or
ampoules. Typically the vials or ampoules are made from glass, e.g.
borosilicate or soda-lime glass. The vials or ampoules may be of
any volume conventional in the art, preferably they are of a size
sufficient to accommodate 0.5 to 5 ml of formulation. The
formulation is stable for periods of storage of up to 12 to 24
months at temperatures of at least 2 to 8.degree. C.
[0181] Formulations must be diluted in an aqueous medium suitable
for intravenous administration before the epothilone can be
administered to a patient.
[0182] The infusion solution preferably must have the same or
essentially the same osmotic pressure as body fluid. Accordingly,
the aqueous medium preferably contains an isotonic agent which has
the effect of rendering the osmotic pressure of the infusion
solution the same or essentially the same as body fluid.
[0183] The isotonic agent may be selected from any of those known
in the art, e.g. mannitol, dextrose, glucose and sodium chloride.
Preferably the isotonic agent is glucose or sodium chloride. The
isotonic agents may be used in amounts which impart to the infusion
solution the same or essentially the same osmotic pressure as body
fluid. The precise quantities needed can be determined by routine
experimentation and will depend upon the composition of the
infusion solution and the nature of the isotonic agent. Selection
of a particular isotonic agent is made having regard to the
properties of the active agent.
[0184] The concentration of isotonic agent in the aqueous medium
will depend upon the nature of the particular isotonic agent used.
When glucose is used it is preferably used in a concentration of
from 1 to 5% w/v, more particularly 5% w/v. When the isotonic agent
is sodium chloride it is preferably employed in amounts of up to 1%
w/v, in particular 0.9% w/v.
[0185] The infusion formulation may be diluted with the aqueous
medium. The amount of aqueous medium employed as a diluent is
chosen according to the desired concentration of Epothilone in the
infusion solution. Preferably the infusion solution is made by
mixing a vial or ampoule of infusion concentrate afore-mentioned
with an aqueous medium, making the volume up to between 20 ml and
200 ml, preferably between about 50 and about 100 ml, with the
aqueous medium.
[0186] Infusion solutions may contain other excipients commonly
employed in formulations to be administered intravenously.
Excipients include antioxidants.
[0187] Antioxidants may be employed to protect the epothilone
against oxidative degradation. Antioxidants may be chosen from any
of those antioxidants known in the art and suitable for intravenous
formulations. The amount of antioxidant may be determined by
routine experimentation. As an alternative to the addition of an
antioxidant, or in addition thereto, the antioxidant effect may be
achieved by displacing oxygen (air) from contact with the infusion
solution. This may be conveniently carried out by purging the
container holding said infusion solution with an inert gas, e.g.
nitrogen.
[0188] Infusion solutions may be prepared by mixing an ampoule or
vial of the formulation with the aqueous medium, e.g. a 5% w/v
glucose solution in WFI or especially 0.9% sodium chloride solution
in a suitable container, e.g. an infusion bag or bottle.
[0189] The infusion solution, once formed, is preferably used
immediately or within a short time of being formed, e.g. within 6
hours.
[0190] Containers for holding the infusion solutions may be chosen
from any conventional container which is non-reactive with the
infusion solution. Glass containers made from those glass types
afore-mentioned are suitable although it may be preferred to use
plastics containers, e.g. plastics infusion bags.
[0191] Plastics containers may be principally those composed of
thermoplastic polymers. Plastics materials may additionally
comprise additives, e.g. plastizisers, fillers, antioxidants,
antistatics and other additives conventional in the art.
[0192] Plastics suitable for the present invention should be
resistant to elevated temperatures required for thermal
sterilisation. Preferred plastics infusion bags are those made from
PVC plastics materials known in the art.
[0193] A wide range of container sizes may be employed. When
selecting a container size, consideration may be paid to the
solubility of the epothilone in the aqueous medium and the ease of
handling and, if appropriate, storage of the container.
[0194] It is preferred to use containers which can accommodate
between about 250 to 1000 ml of infusion solution, but preferably
about 50 to about 120 ml.
[0195] Infusion solutions act in a similar fashion to infusion
solutions of the microtubule interacting agent paclitaxel, and are
beneficial in treating conditions for which paclitaxel might be
used. For certain tumors epothilones offer enhanced beneficial
effects compared with paclitaxel.
[0196] Dosage forms may be conveniently administered intravenously
in a dosage of up to 100 mg/m.sup.2 Epothilone A and up to about 18
mg/m.sup.2 of Epothilone B. The exact dosage required and the
duration of administration will depend upon the seriousness of the
condition and the rate of administration, and it is preferably as
defined above. As the dose may be delivered intravenously, the dose
received and the blood concentration can be determined accurately
on the basis of known in vivo and in vitro techniques.
[0197] Pharmaceutical compositions for oral administration can be
obtained by combining the active ingredient with solid carriers, if
desired granulating a resulting mixture, and processing the
mixture, if desired or necessary, after the addition of appropriate
excipients, into tablets, dragee cores or capsules. It is also
possible for them to be incorporated into plastics carriers that
allow the active ingredients to diffuse or be released in measured
amounts.
[0198] Suitable pharmaceutically acceptable carriers are especially
fillers, such as sugars, for example lactose, saccharose, mannitol
or sorbitol, cellulose preparations and/or calcium phosphates, for
example tricalcium phosphate or calcium hydrogen phosphate, and
binders, such as starch pastes using for example corn, wheat, rice
or potato starch, gelatin, tragacanth, methylcellulose,
hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or
polyvinylpyrrolidone, and/or, if desired, disintegrators, such as
the above-mentioned starches, also carboxymethyl starch,
crosslinked polyvinylpyrrolidone, agar, alginic acid or a salt
thereof, such as sodium alginate. Excipients are especially flow
conditioners and lubricants, for example silicic acid, talc,
stearic acid or salts thereof, such as magnesium or calcium
stearate, and/or polyethylene glycol. Drage cores are provided with
suitable, optionally enteric, coatings, there being used, inter
alia, concentrated sugar solutions which may comprise gum arabic,
talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium
dioxide, or coating solutions in suitable organic solvents, or, for
the preparation of enteric coatings, solutions of suitable
cellulose preparations, such as ethylcellulose phthalate or
hydroxypropylmethylcellulose phthalate. Capsules are dry-filled
capsules made of gelatin and soft sealed capsules made of gelatin
and a plastiziser, such as glycerol or sorbitol. The dry-filled
capsules may comprise the active ingredient in the form of
granules, for example with fillers, such as lactose, binders, such
as starches, and/or glidants, such as talc or magnesium stearate,
and if desired with stabilizers. In soft capsules the active
ingredient is preferably dissolved or suspended in suitable oily
excipients, such as fatty oils, paraffin oil or liquid polyethylene
glycols, and also stabilizers and/or antibacterial agents may be
added. Dyes or pigments may be added to the tablets or dragee
coatings or the capsule casings, for example for identification
purposes or to indicate different doses of active ingredient.
[0199] In the case of combinations with an other chemotherapeutic,
a fixed combination of two or more components (a) and (b) as
defined above or two or more independent formulations (e.g. in a
kit of part) are prepared as described above, or the other
chemotherapeutic(s) is/are used in standard formulations that are
marketed and known to the person of skill in the art.
EXAMPLES
[0200] The following Examples are intended to illustrate the
present invention but are not intended to limit the scope thereof.
Especially the cell lines mentioned therein are not thought to
limit the scope of the invention as they are merely representatives
and may be replaced with different cell lines and tumor cells for
which they are representatives.
[0201] Preparation of compound solutions
[0202] A stock solution of epothilone B at 10 mg/ml in DMSO is
prepared and stored at -20.degree. C. Aliquots are diluted in
aqueous solutions to a final concentration of 5% v/v DMSO, 0.05%
v/v Tween 80 (polyoxyethylen-sorbitan-monooleate; ICI Americas,
Inc.), and 95% v/v physiological saline (0.9% w/v NaCl).
[0203] Cells and Cell Culture Conditions
[0204] Human colorectal adenocarcinoma cell line HCT-15 (ATCC CCL
225) is from the American Type Culture Collection (Rockville, Md.,
USA), and the cells are cultivated in vitro as recommended by the
supplier. HCT-15 is an epithelial-like cell line (Cancer Res. 39:
1020-25 [1979]) that is multi-drug resistant by virtue of
over-expression of P-glycoprotein (P-gp, gp170, MDR-1; Anticancer
Res. 11: 1309-12 [1991]; J. Biol. Chem. 264: 18031-40 [1989]; Int.
J. Cancer 1991; 49: 696-703 [1991]) and glutathione-dependent
resistance mechanisms (Int. J. Cancer 1991; 49: 688-95.[1991]).
[0205] The Colo 205 cell line is also a human colon carcinoma cell
line (ATCC No. CCL 222; see also Cancer Res. 38, 1345-55 [1978]
which was isolated from ascitic fluid of a patient, displays
epithelial-like morphology and is generally considered to be
drug-sensitive.
[0206] A human androgen-independent prostate cancer cell line is
used to establish subcutaneous and orthotopic models in mice. The
human metastatic prostate carcinoma PC-3M is obtained from Dr. I.
J. Fidler (MD Anderson Cancer Center, Houston, Tex., USA) and is
cultured in Ham's F12K media supplemented with 7% v/v FBS. The
PC-3M cell line is the result of isolation from liver metastasis
produced in nude mice subsequent to intrasplenic injection of PC-3
cells [ATCC CRL 1435; American Type Culture Collection (Rockville,
Md., USA)], and they can grow in Eagle's MEM supplemented with 10%
fetal bovine serum, sodium pyruvate, non-essential amino acids,
L-glutamine, a two-fold vitamin solution (Gibco Laboratories, Long
Island, N.Y.) and penicillin-streptomycin (Flow Laboratories,
Rockville, Md.). The PC-3M cell line is hormone-insensitive (that
is, it grows in the absence of androgens). The PC-3 cell line is
androgen receptor negative, as is presumably the derived PC-3M cell
line. PC-3 is a cell line available from ATCC (ATCC CRL 1435) and
corresponds to a grade IV prostatic adenocarcinoma isolated from a
62-year-old Caucasian male; the cells exhibit low acid phosphatase
and testosterone-5-.alpha.-reductase activity. The cells are
near-triploid with a modal number of 62 chromosomes. No normal Y
chromosomes can be detected by Q-band analysis.
[0207] Human lung adenocarcinoma A549 (ATCC CCL 185; isolated as
explant culture from lung carcinoma tissue from a 58-year-old
Caucasian male); shows epithelial morphology and can synthesize
lecithin with a high percentage of desaturated fatty acids
utilizing the cytidine diphosphocholine pathway; a subtelocentric
marker chromosome involving chromosome 6 and the long arm of
chromosome 1 is found in all metaphases. The human breast carcinoma
ZR-75-1 (ATCC CRL 1500; isolated from a malignant ascitic effusion
of a 63-year-old Caucasian female with infiltrating ductal
carcinoma); is of mammary epithelial origin; the cells possess
receptors for estrogen and other steroid hormones and have a
hypertriploid chromosome number.
[0208] The human epidermal (mouth) carcinoma cell line KB-8511 (a
P-gp over-expressing cell line derived from the epidermoid (mouth)
KB-31 carcinoma cell line) is obtained from Dr. R. M. Baker,
Roswell Park Memorial Institute (Buffalo, N.Y., USA) (for
description see Akiyama et al., Somat. Cell. Mol. Genetics 11,
117-126 (1985) and Fojo A., et al., Cancer Res. 45, 3002-3007
(1985)) and is cultured as previously described (Meyer, T., et al.,
Int. J. Cancer 43, 851-856 (1989)): KB-8511 cells, like KB-31, are
derived from the KB cell line (ATCC) and they are human epidermal
carcinoma cells; KB-31 cells can be grown in mono-layer using
Dulbecco's modified Eagle's medium (D-MEM) with 10% fetal calf
serum (M.A. Bioproducts), L-glutamine (Flow), penicillin (50
units/ml) and streptomycin (50 .mu.g/ml (Flow); they then grow with
a doubling time of 22 h, and their relative plating efficiency is
approximately 60%. KB-8511 is a cell line derived from the KB-31
cell line by use of colchicine treatment cycles; it shows about a
40-fold relative resistance against colchicine when compared with
the KB-31 cells; it can be grown under the same conditions as
KB-31.
[0209] For more details on the characteristics of the cell lines,
see the ATCC catalogue and references cited therein, or the other
references cited above.
[0210] The following cell lines mentioned above have been deposited
under the Budapest Treaty on Feb. 20, 1998, at the Deutsche
Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ,
Mascheroder Weg 1 b, D-38124 Braunschweig, Germany) under the
following accession numbers, respectively: PC-3M: DSM ACC2338;
A-549: DSM ACC2337; KB-8511: DSM ACC2342.
[0211] In addition, the following cell lines mentioned above have
been deposited under the Budapest Treaty on Dec. 1, 1998, at the
Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ,
Mascheroder Weg 1b, D-38124 Braunschweig, Germany) under the
following accession numbers, respectively: ZR-75-1: DSM ACC2376;
HCT-15: ACC2377.
[0212] In the following, general methods are described for the
tests made. Where specific conditions are mentioned, these prevail
over the general descriptions presented in the next paragraphs:
[0213] Antiproliferative Assays:
[0214] Antiproliferative assays are performed as previously
described (Int. J. Cancer 43, 851-6 (1989)). Briefly, cells are
seeded at 1.5.times.10.sup.3 cells/well into 96-well microtiter
plates and incubated overnight. Compounds are added in serial
dilutions on day 1. The plates are then incubated for an additional
2 to 5 days, allowing for at least one cell doubling (cell line
dependent), after which the cells are fixed with 3.3% v/v
glutaraldehyde, washed with water and stained with 0.05% w/v
methylene blue. After washing, the dye is eluted with 3% V/V HCl
and the optical density measured at 665 nm with a SpectraMax 340
(Bucherer, Basel, Switzerland). IC50 values are determined by a
computerized system (SoftPro, Bucherer, Basel, Switzerland) using
the formula (OD test-OD start)/(OD control-OD start).times.100.
IC50 is defined as the drug concentration which leads to 50% of
cells per well compared to control cultures (100%) at the end of
incubation period.
[0215] In Vivo Antitumor Activity Against s.c. Transplanted
Tumors
[0216] Female or male BALB/c nu/nu (nude) mice are kept under
sterile conditions (10 to 12 mice per Type III cage) with free
access to food and water. Mice weigh between 20 and 25 grams at the
time of tumor implantation. Tumors are established by subcutaneous
injection of cells (minimum 2.times.10.sup.6 cells in 100 .mu.l PBS
or medium) in carrier mice (4-8 mice per cell line). The resulting
tumors are serially passaged for a minimum of three consecutive
transplantations prior to start of treatment. Tumor fragments
(approx. 25 mg) are implanted s.c. into the left flank of animals
with a 13-gauge trocar needle while the mice are exposed to Forene
(Abbott, Switzerland) anesthesia.
[0217] Tumor growth and body weights are monitored once or twice
weekly. All treatments are administered intravenously (i.v.) and
are initiated when a mean tumor volume of approximately 100 to 250
mm.sup.3 is attained, depending upon the tumor type. Tumor volumes
are determined using the formula (L.times.D.times..pi.)/6 (see
Cancer Chemother. Pharmacol. 24:148-154, [1989]). Treatments with
epothilone B vary the dose and the frequency of administration.
Comparator agents are administered according to previously
determined optimal treatment regimens. In addition to presenting
changes in tumor volumes over the course of treatment, antitumor
activity is expressed as T/C% (mean increase of tumor volumes of
treated animals divided by the mean increase of tumor volumes of
control animals multiplied by 100). Tumor regression (%) represents
the smallest mean tumor volume compared to the mean tumor volume at
the start of treatment, according to the formula Regression
(%)=(1-V.sub.end/V.sub.start).times.100 (V.sub.end=final mean tumor
volume, V.sub.start=mean tumor volume at the start of treatment.
Any animals in which the tumor has reached a size exceeding
approximately 1.5 to 2.5 cm.sup.3 are sacrificed. Details can be
found below.
[0218] In Vivo Antitumor Activity Against Orthotopically Injected
Cells
[0219] Human prostate carcinoma PC-3M cells (1.times.10.sup.6 cells
per 20 .mu.l phosphate buffered saline) are injected into the left
ventricle of the prostate of each animal (n=6-9/group) according
the method previously described (see Stephenson et al., J. Natl.
Cancer Inst. 84, 951-7 (1992)). Treatment is started on day 14
after cell injection. At this time point a mean tumor weight of
.about.20 mg is established. Epothilone B is administered either
once or once weekly. Mice are sacrificed 42 days post tumor
inoculation, and the prostates are carefully removed, dissected
free of adhering tissue and weighed. Mesengeal lymph nodes are
examined for the presence of metastases, dissected free of adhering
tissue and weighed. Mesenteric lymph nodes are examined for the
presence of metastases, dissected free of adhering tissue and
weighed.
[0220] Tumor weight and body weights are monitored once or twice
weekly. All treatments are administered intravenously (i.v.) and
are initiated 14 days post cell injection. Treatment with
epothilone B varies the dose and frequency of administration.
Antitumor activity is expressed as T/C% (mean tumor weight of
treated animals divided by the mean tumor weight of control animals
multiplied by 100).
[0221] Statistical Analyses
[0222] The basic strategy for statistical analyses is to use tests
for multiple comparisons to judge the statistical significance of
differences between treatment groups, and differences within a
group (i.e. comparing to the start of treatment) to determine if
treatment induces stable disease or tumor regressions. As
subcutaneous tumor volumes are not normally distributed,
differences in the subcutaneous tumor volumes between treatment
groups are determined using the non-parametric Kruskal-Wallis one
way ANOVA test on ranked data (Rank sum test), and the statistical
significance of differences between treatment groups as compared to
control groups determined using the Dunnett test. Pair-wise
comparisons between all groups is performed using the
Student-Newman-Keuls (SNK) method. Organ weights are not always
normally distributed and are analyzed either using non-parametric
tests (as above) or are transformed to normality (Log [organ
weight]) and are analyzed by One-way ANOVA followed by the Dunnett
test (comparisons to controls) or using Tukey (between group
comparisons). Statistical analysis on A tumor volumes uses the
Kruskal-Wallis one way ANOVA test on ranked data, comparing
treatment groups with vehicle controls by Dunnett's test.
Differences in the body weights between treatment groups with
vehicle controls are analyzed by paired t-tests. Fisher's Exact
Test is used to determine the significance of differences in
mortalities between the groups. For all tests the level of
significance is set at p<0.05, but note that for these small
sample sizes, the desired power level of 0.8 is never obtained. For
the orthotopic model, Fisher's exact test is used to determine
whether the ratios of mice bearing metastases are different between
treatment groups and controls. Any statistical calculations are
performed using SigmaStat 2.0 (Jandel Scientific).
[0223] Material
[0224] Epothilone B is purified from cultures of the myxobacterium
Sorangium cellulosum by Biopharmaceuticals Production and
Development Department, Novartis Pharma, Basel, Switzerland.
TAXOL.RTM. (paclitaxel formulated for clinical use) is purchased
from Bristol Myers Squibb (USA), paclitaxel from Calbiochem (USA),
and 5-fluorouracil (Fluoro-uracil) is from Roche (Switzerland).
Cell culture materials are from Integra BioSciences (Wallisellen,
Switzerland). Liquid media, fetal bovine serum (FBS) and media
additives are from Gibco/BRL (Basel, Switzerland). BALB/c nu/nu
(nude) mice are from Bomholtgaard (Copenhagen, Denmark) or obtained
from the Novartis Animal Farm (Sisseln, Switzerland). Normal BALB/c
mice are from Iffa Credo (France) or obtained from Novartis Animal
Farm (Sisseln, Switzerland).
[0225] Determination of the Maximal Tolerated Dose (MTD)
[0226] For the MTD determination, female BALB/c nude mice or BALB/c
mice (obtained from Novartis Animal Farm, Sisseln, Switzerland) are
treated once intravenously with epothilone B (n=3 per dose group).
The dosage is increased (2, 4, 6, 8, 10 and 12 mg/kg) and the mice
are observed for the presentation of overt toxic effects for 10
days after drug treatment.
Example 1
Determination of the Maximal Tolerated Dose (MTD)
[0227] The results from the maximal tolerated dose (MTD) study are
presented in Table 1 and Table 2.
1TABLE 1 Determination of the singl i.v. dose MTD for pothilone B
in normal female BALB/c mice. Body Weight (mean g .+-. SD) Dose
.DELTA.(p value) % Change Mortalities Experiment 1 12 -5.4 .+-. 2.9
-23.6 .+-. 11.9 3/3 (p = 0.085) 10 -5.6 .+-. 0.6 -24.9 .+-. 1.6 3/3
(p = 0.003) 8 -5.1 .+-. 3.2 -22.1 .+-. 13.1 3/3 (p = 0.110) 6 -6.1
.+-. 0.9 -28.6 .+-. 3.5 1/3 (p = 0.007) Experiment 2 8 -5.0 .+-.
1.0 -25.3 .+-. 3.9 3/3 (p = 0.013) 6 -5.3 .+-. 0.6 -27.5 .+-. 2.1
2/3 (p = 0.004) 4 1.3 .+-. 0.6 6.5 .+-. 2.6 0/3 (p = 0.057) 2 2.3
.+-. 0.6 12.3 .+-. 3.0 0/3 (p = 0.02)
[0228] Epothilone B is given as a single i.v. bolus dose at 2, 4,
6, 8, 10 or 12 mg/kg. Survival and body weights of the mice are
monitored daily. Changes (.DELTA.) in body weights are determined
comparing the last measured body weight to that before
treatment.
2TABLE 2 Determination of the single i.v. dose MTD for epothilone B
in female BALB/c nude mice. Body Weight (mean g .+-. SD) Dose
.DELTA.(p value) % Change Mortalities 12 -5.1 .+-. 0.6 -22.9 .+-.
3.1 3/3 (p = 0.004) 10 -5.6 .+-. 3.3 -23.7 .+-. 13.6 1/3 (p =
0.336) 8 -3.8 .+-. 2.7 -16.8 .+-. 12.2 1/3 (p = 0.250) 6 -1.0 .+-.
0.5 -4.2 .+-. 2.1 0/3 (p = 0.077) 4 -0.4 .+-. 0.6 -1.7 .+-. 2.9 0/3
(p = 0.427) 2 1.0 .+-. 0.5 4.2 .+-. 2.1 0/3 (p = 0.071) 0 0.6 .+-.
0.6 3.0 .+-. 2.9 0/3 (p = 0.151)
[0229] Epothilone B is given as a single i.v. bolus dose at 2, 4,
6, 8, 10 or 12 mg/kg. Survival and body weights of the mice are
monitored daily. Changes (A) in body weights are determined
comparing the last measured body weight to that before
treatment.
[0230] If follows from these experiments that in normal mice the
MTD is around 4 mg/kg, while in nude mice the MTD is around 6
mg/kg.
Example 2
Toxicity of Epothilone B (Two-Week Intravenous Comparative Toxicity
Study in Mice)
[0231] In order to assess the sub-chronic intravenous toxicity of
epothilone B, a non-GLP two-week i.v. toxicity study in non-tumor
bearing, normal female BALB/c mice is conducted, involving analysis
of mortality, clinical signs, body weight, food consumption,
hematology, clinical biochemistry, urinalysis, and organ weights as
well as macro and microscopic examinations. The study is based on
two different dosing regimens of epothilone B of 3 mg/kg and 10
mg/kg, respectively, administered on days 1 and 8 (8 animals per
group). Half of the animals are killed on day 15 (main group) and
necropsy is performed. For the other half (recovery group) a
recovery period of 5 weeks is allowed after administration of the
second dose before sacrifice and subsequent necropsy on day 43.
However, for the 10 mg/kg dose all animals of the recovery group
have to be sacrificed prematurely on day 19, due to poor general
condition.
[0232] No mortalities occur throughout the treatment period at
either dose level (days 1-14) and for the 3 mg/kg dose group all
animals of the recovery group survive until the end of the study.
Body weight loss is observed for all animals in the 10 m/kg dose
group during the first and second week, whereas body weight loss at
the 3 mg/kg dose is only apparent in the second week. Body weight
development during the recovery period is similar for treated and
control animals.
[0233] Both dose levels of epothilone B induce a reduction in the
number of leukocytes, especially of neutrophils and lymphocytes, in
all treated animals (day 15), but the effect is more pronounced at
the 10 mg/kg dose. In addition, slight increases in basophil counts
as well as decreased levels of monocytes are observed in some
individual animals at both dose levels. Slightly lower values for
red blood cell parameters with increases in reticulocytes and
platelets are recorded only for the 10 mg/kg dose. At the end of
the recovery period (day 43) hematological parameters are normal
for two out of the four animals of the 3 mg/kg dose recovery group,
whereas the other two still suffer from reduced white blood cell
counts.
[0234] Only minor changes are observed in the clinical chemistry
profile of treated animals, which cannot be clearly related to
treatment with epothilone B.
[0235] Treatment with epothilone B at both dose levels leads to
pronounced changes in thymus, spleen, and uterus weights (day 15).
In addition, slight reductions in liver weight are observed. (Organ
weight is determined for adrenal glands, liver, thymus, spleen,
brain, ovaries, kidneys, uterus, and heart). For the 3 mg/kg dose,
organ weights at the end of the recovery period (day 43) are
comparable to those of control animals, indicating full recovery.
(No organ weights are taken for the sacrificed animals of the 10
mg/kg dose recovery group).
[0236] Microscopic investigation of histologically processed
selected tissues from animals sacrificed on day 15 reveals moderate
to marked atrophy of the thymus for the 3 mg/kg and the 10 mg/kg
dose, respectively. In addition, minimal lymphoid atrophy in the
spleen, minimal to slight myeloid hypoplasia in the sternal bone
marrow, and minimally increased hemopoiesis in the spleen are
observed at 10 mg/kg dose. At 3 mg/kg the sternal bone marrow shows
minimal to slight erythroid and myeloid atrophy. Minimal single
cell necrosis is noted in the intestinal mucosa (small and large
intestine) at both dose levels, but with a higher incidence at the
10 mg/kg dose.
[0237] Animals from the recovery groups at both dose levels show
slight myeloid hyperplasia and/or athropy in the bone marrow. At 10
mg/kg slight lymphoid atrophy is also seen in the spleen and in
addition slight to moderate hemosiderosis is present. There is no
thymic tissue available for microscopic examination at 10 mg/kg,
indicating that thymus atrophy might also have been present in
these mice. No histological alterations in the thymus are found on
day 43 for the animals from the 3 mg/kg dose recovery group.
[0238] In conclusion, at a dose level of 3 mg/kg (dosing on days 1
and 8) epothilone B is tolerated without mortality for the total
observation period of 43 days, while animals dosed with 10 mg/kg
have to be sacrificed on day 19, due to poor health condition. Body
weight loss occurs at both dose levels during the treatment period.
Hematology reveals lower values for leukocytes, neutrophils and
lymphocytes with higher basophil and lower monocyte counts for some
individual mice in both dose groups. In addition, evidence of
anemia is seen at both dose levels. No effects on clinical
chemistry profile are observed. Moderate to marked atrophy of the
thymus are observed at 3 and 10 mg/kg after the treatment period
only (day 15), together with minimal lymphoid atrophy in the spleen
at the 10 mg/kg dose. In addition, myeloid hypoplasia in bone
marrow and increased hemopoiesis in the spleen are detectable at 10
mg/kg. Slight erythroid and myeloid atrophy of bone marrow is seen
at 3 mg/kg. Single cell necrosis is detected in the intestinal
mucosa at 3 and 10 mg/kg at the end of the treatment period
only.
[0239] Conclusions
[0240] The most important conclusions emerging from the data
summarized in Examples 1 and 2 for the toxicological findings with
epothilone B can be summarized as follows:
[0241] The MTD for single dose i.v. administration of epothilone B
to normal and nude mice from a BALB/c background corresponds to 4
mg/kg and 6 mg/kg, respectively. Nude mice thus are less sensitive
to the toxic effects of the compound than normal mice.
[0242] In normal mice, two doses of 3 mg/kg given one week apart
are reasonably well tolerated and do not cause mortality up to day
43 after the initial dose. The same dosing regimen at a 10 mg/kg
dose level results in death (or sacrifice) of all treated
animals.
Example 3
In Vitro Activity of Epothilones Against Cell Lines
[0243] The potent anti-proliferative activity of epothilone B is
confirmed for some human cancer cell lines; the results of these
experiments are summarized in Table 3. Epothilone B generally
exhibits higher potency than paclitaxel, particularly against
cancer cells with a multidrug resistant (MDR) phenotype (e.g.
KB-8511, HCT-15).
3TABLE 3 In vitro activity of the epothilones against human
carcinoma cell lines. IC50-values [nM] for growth inhibition of
human carcinoma cell lines by epothilone B in comparison to
paclitaxel (5 d exposure, mean .+-. SD, n = 3). Values in
parenthesis indicate relative resistance, i.e., IC50 (resistant
line)/IC50 (parental line). Cell Line epothilone B paclitaxel A549
(Lung) 0.19 .+-. 0.12.sup.a 3.75 .+-. 0.92 ZR-75-1 (Breast) 0.64
.+-. 0.42.sup. 3.60 .+-. 1.87 HCT-15 (Colon) 0.41 .+-. 0.15.sup.
106 .+-. 54 KB-8511 0.89 .+-. 0.47.sup.b .sup. 994 .+-. 281.sup.b
(Epidermoid, MDR) (1.25) (343).sup.a PC-3M (Prostate) 3.82 .+-.
0.47.sup.c .sup. 6.74 .+-. 0.72.sup.c .sup.aMean .+-. SD, n = 2.
.sup.b2 d Exposure; .sup.c3 d exposure MDR = Multidrug
Resistant
Example 4
Antitumor Activity of the Epothilones Against Human Colorectal
Adenocarcinoma HCT-15 Tumors
[0244] Tumor volumes are used as the primary indicator of activity
of antitumor agents used alone or in combination, and changes in
body weights are measured as an indicator of treatment
tolerability.
[0245] As can be deduced from Table 4, a single 4 mg/kg dose of
epothilone B is able to produce tumor regressions (p<0.05 vs.
vehicle controls; Dunnett's) in drug-resistant, P-gp
over-expressing, HCT-15 colon tumors (FIG. 1 and Table 1). This
activity is clearly superior to five 20 mg/kg administrations of
TAXOL.RTM. or two 75 mg/kg 5-fluorouracil administrations
(p<0.05 vs. epothilone B; SNK test). HCT-15 tumors are resistant
to both TAXOL.RTM. and 5-fluorouracil, in that final TIC values of
50% and 88%, respectively, are obtained (both p>0.05 vs.
controls; Dunnett's). Epothilone B treatment is well tolerated in
that body weight is stable under treatment; vehicle-treated mice
gain weight. No mortalities due to treatment are observed with
epothilone B. In contrast, some mortalities are observed with
TAXOL.RTM. treatment ({fraction (1/8 )} [12.5%] deaths) and a
greater extent of lethality occurs with 5-fluorouracil ({fraction
(4/8)} [50%] deaths); however, due to the small size of the
treatment groups, this does not reach statistical significance
(p>0.05 vs. controls; Fisher's Exact Test). Mice surviving
either treatment demonstrate stable body weights.
[0246] This result indicates epothilone B produces a pronounced
anti-tumor effect against HCT-15 tumors resistant to TAXOL.RTM. and
5-fluorouracil and is well-tolerated at this 4 mg/kg dose.
4TABLE 4 Antitumor ffect of epothilone B in compari on with TAXOL
.RTM. or 5-fluorouracil against subcutaneously transplanted human
HCT- 15 colon carcinoma in female BALB/c nude mice. Host Response
Tumor Response Survival Dose, D Tumor D Body % Body (No. Com-
Route, Regres- Volume Weight Weight alive/ pound Schedule T/C sion
(mm.sup.3) (g) Change total) Vehicle 25 ml/kg, 100% none 1939 .+-.
2.1 .+-. 0.5 10 .+-. 2 8/8 con-trols iv. 333 (p = days 14, 0.003)
16, 18, 20 and 22 epothi 4 mg/kg, Regres- -61% -97 .+-. 25 0.4 .+-.
0.3 2 .+-. 2 8/8 lone B i.v. sion (p < (ns) once on 0.05) day 14
5-fluoro- 75 mg/kg, 88% none 1654 .+-. 2.0 .+-. 0.7 9 .+-. 4 4/8
uracil i.v., on 824 (ns) days 14 (ns) and 21 TAXOL .RTM. 20 mg/kg,
50% none 963 .+-. 0.7 .+-. 0.6 3 .+-. 3 7/8 i.v. 298 (ns) days 14,
(ns) 16, 18, 20 and 22
[0247] Tumor fragments of approximately 25 mg are implanted into
the left flank of each female nude mouse (n=8 per group).
Treatments are started on day 14 after tumor transplantation.
Epothilone B is administered once at 4 mg/kg, i.v. on day
14.5-Fluorouracil is administered at 75 mg/kg, i.v., on days 14 and
21. TAXOL.RTM. is administered i.v. at 20 mg/kg/day, every second
day for 5 treatments (days 14, 16, 18, 20 and 22). Antitumor
activity is expressed as T/C% (mean increase of tumor volumes of
treated animals divided by the mean increase of tumor volumes of
control animals multiplied by 100). Tumor regression (%) represents
the final mean tumor volume compares to the mean tumor volume at
the start of treatment. .DELTA. Tumor volumes represent the tumor
volume on the last treatment day minus the tumor volume on the
first treatment day. Statistical analyses on .DELTA. tumor volumes
uses Dunnett's test to compare treatment groups to controls.
Statistical analyses on body weight changes uses paired t-tests
comparing body weights before treatment to those at the end of
treatment; mice weigh .about.20-25 g at the start of treatment. Not
significant is indicated by the abbreviation ,,ns". Data presented
are means.+-.SEM from animals surviving to the end of the
experiment.
Example 5
Antitumor Effect of Epothilone B in Comparison with TAXOL.RTM.
Against Subcutaneously Transplanted Human KB-8511 Epidermoid
Carcinoma in Female BALB/c Nude Mice:
[0248] As can be deduced from Table 5, various regimens of
epothilone B are able to inhibit the growth of TAXOL.RTM.-resistant
KB-8511 tumors in nude mice. A single administration of 4 mg/kg
epothilone B produces a transient regression (-51% on day 25 post
transplantation; p<0.05 vs. vehicle controls, Dunnett), but the
tumors re-grows by day 40 post-treatment to result in a final T/C
of 24% (p<0.05 vs. vehicle controls, Dunnett). This single
epothilone B administration is well tolerated producing stable body
weights, and no mortalities occur.
[0249] Once weekly intravenous administration of epothilone B
results in dose-dependent inhibition of tumor growth: 4 mg/kg
produces 98% regressions (p<0.05 vs. vehicle controls; Dunnett);
2 mg/kg, produces transient 44% regressions and a final T/C of 14%
(both p<0.05 vs. vehicle controls; Dunnett); 1 mg/kg produces a
final T/C 81% (p>0.05 vs. vehicle controls; Dunnett). TAXOL.RTM.
is inactive against KB-8511 tumors (T/C 132%, p>0.05 vs. vehicle
controls; Dunnett). At the end of the experiment, {fraction (5/8 )}
mice treated with 4 mg/kg/week, and {fraction (1/8 )} mice treated
with 2 mg/kg/week epothilone B have undetectable tumors. Although 4
mg/kg once per week tend to reduce body weights (-5.+-.7%), this
does not reach statistical significance. Vehicle controls, 2 and 1
mg/kg/week epothilone B, and TAXOL.RTM. groups all display
increasing body weights, and there are no mortalities, suggesting
well-tolerated treatments.
[0250] These results indicate that epothilone B is effective
against experimental epidermoid tumors that are
TAXOL.RTM.-resistant.
5TABLE 5 Antitumor effect of epothilone B in comparison with TAXOL
.RTM. against subcutaneously transplanted human KB-8511 epidermoid
carcinoma in female BALB/c nud mic. Host Response Tumor Response
Survival Dose, D Tumor D Body % Body (No. Com- Route, Regres-
Volume Weight Weight alive/ pound Schedule T/C sion (mm.sup.3) (g)
Change total) Vehicle 25 100% none 2001 .+-. 2.6 .+-. 0.3 12 .+-. 2
8/8 con-trols ml/kg, 405 (p < i.v. 0.001) once per week epothi-
4 mg/kg, 24% -51% 484 .+-. 0.6 .+-. 0.6 3 .+-. 2 8/8 lone B i.v.
(trans- 103 (ns) once on ient) (p < day 13 0.05) epothi- 4
mg/kg, Regres- -98% -107 .+-. -1.1 .+-. 0.4 -5 .+-. 3 8/8 lone B
i.v. sion 14 (ns) once per (p < week 0.05) epothi- 2 mg/kg, 14%
-44% 289 .+-. 1.1 .+-. 0.4 5 .+-. 2 8/8 lone B i.v. (trans- 204 (p
= once per ient) (p < 0.031) week 0.05) epothi- 1 mg/kg, 81%
none 1620 .+-. 2.6 .+-. 0.3 12 .+-. 1 8/8 lone B i.v. 290 (p = once
per (ns) 0.008) week TAXOL .RTM. 20 132% none 2662 .+-. 3.3 .+-.
0.6 15 .+-. 3 8/8 mg/kg, 509 (p < i.v. (ns) 0.001) days 13, 15,
17, 19 and 21
[0251] Tumor fragments of approximately 25 mg are implanted into
the left flank of each female nude mouse (n=8 per group).
Treatments are started on day 13 after tumor transplantation.
epothilone B is administered once at 4 mg/kg, i.v. on day 13, or
once per week at 4, 2 or 1 mg/kg, i.v., (on days 13, 21 and 27).
TAXOL.RTM. is administered i.v. at 20 mg/kg/day, every second day
for 5 treatments (days 13, 15, 17, 19 and 21). Antitumor activity
is expressed as T/C% (mean increase of tumor volumes of treated
animals divided by the mean increase of tumor volumes of control
animals multiplied by 100). Tumor regression (%) represents the
final mean tumor volume compared to the mean tumor volume at the
start of treatment. Changes (.DELTA.) in Tumor volumes represent
the tumor volume on the last treatment day minus the tumor volume
on the first treatment day. Statistical analyses on .DELTA. tumor
volumes uses Dunnett's test to compare treatment groups to
controls. Statistical analyses on body weight changes use paired
t-tests comparing body weights before treatment to those at the end
of treatment; mice weigh .about.20-25 g at the start of treatment.
Not significant is indicated by the abbreviation ,,ns". Data
presented are means.+-.SEM from animals surviving to the end of the
experiment.
[0252] It follows from the experiment that, while TAXOL.RTM. is not
effective, epothilone B treatment shows effective antitumor
activity; even regression can be found at the 4 mg/kg dose.
Example 6
Antitumor Activity of the Epothilones Against Orthotopically
Injected PC-3M Prostate cells.
[0253] The results determining the activity of epothilone B against
PC-3M tumors initially growing in prostate and then forming
metastases in mesengeal lymph nodes are presented in Table 6.
[0254] In this experimental prostate cancer model, PC-3M cells
initially grow in the prostate and then form metastases in
mesenteric lymph nodes. Organ weights are used to assess antitumor
activity of the treatments.
[0255] Table 6 represents the progress of the experimental cancer
from 14 days post cell injection (start of treatment), where no
lymph node involvement is observed, to 42 days post cell injection,
where prostate and mesenteric lymph nodes dramatically increase in
weight. All administration regimens of epothilone B are highly
effective (all p<0.05 versus controls; Dunnett's test on log
transformed data) in reducing prostate weights and preventing
metastases of the tumor to the mesenteric lymph nodes. All of the
active treatments are equivalent in antitumor activity (p>0.05;
Dunn's). In each of the epothilone B treatment groups, only one
animal has detectable metastases, as compared to all animals in the
vehicle-treated controls (p<0.05; Fisher's Exact test),
indicating that treatment with epothilone B significantly impairs
the formation of detectable metastases.
[0256] Epothilone B treatments are not well-tolerated at the higher
doses. Whereas a single administration of 6 mg/kg, or two
administrations of 4 mg/kg, only tend to reduce body weights,
administration of 8 mg/kg once, or 5 mg/kg once weekly, produces
significant losses in body weight (Table 6). Treatment appears to
promote survival of tumor-bearing mice; however, likely owing to
the small numbers per treatment group, this only reaches
statistical significance with the 6 mg/kg (once) regimen (p=0.029,
Fisher's Exact test on final survival numbers).
[0257] Epothilone B demonstrates excellent antitumor activity in
this model, both in terms of reduction of primary tumors and
prevention of metastases. However, epothilone B, in some regimens,
is poorly tolerated.
[0258] The results of this study indicate that epothilone B is
active against human prostate carcinomas both in vitro and in vivo
(see Example 3). Epothilone B is able to reduce the growth of the
primary tumor and potently inhibit the formation of detectable
metastases in an orthotopic model of prostate cancer. Furthermore,
it may also promote survival of these tumor-bearing mice, although
this needs to be examined in additional experiments. In parallel
with its potent antitumor activity, epothilone B treatment produces
significant body weight losses with the tested dosage regimens.
However, the reasons for this poor tolerability are unknown.
[0259] The activity of epothilone B in the orthotopic PC-3M model
is especially noteworthy. Orthotopic models are designed to implant
the tumor within the tissues where the primary tumor is located in
humans, and unlike most subcutaneous tumor implantation models,
metastases frequently arise. Therefore, the repression of primary
tumor growth in the prostate by epothilone B, and the inhibition of
the formation and/or growth of mesenteric lymph node metastases
represents a significant activity of epothilone B.
[0260] In summary, owing to potent antitumor activity in
experimental prostate cancer models, which are considered
relatively resistant to chemotherapy (Br. J. Cancer 75, 1593-600
(1997)), epothilone B appears to be a promising agent for the
treatment of prostate cancer.
6TABLE 6 Antitumor effect of epothilone B against orthotopically
injected human PC-3M prostate carcinoma cells in male BALB/c nude
mice. Tumor Response Host Response Dose, (T/C %) .DELTA. Body %
Body Survival Com- Route Lymph Weight Weight (No. pound Schedule
Prostate Nodes (g) Change alive/total) Vehicle 25 100% 100% -1.6
.+-. -5 .+-. 9 4/9 con- ml/kg, 1.8 trols i.v. (ns) once Epothi- 8 1
4 -5.2 .+-. -13 .+-. 15 7/9 lone B mg/kg, 4.3 i.v. (p = once 0.009)
6 5 4 -1.4 .+-. -6 .+-. 12 9/9 mg/kg, 4.1 i.v. (ns) once 5 1 4 -6.8
.+-. -25 .+-. 5 7/9 mg/kg, 1.5 i.v. (p < once 0.001) per week 4
2 4 -1.2 .+-. -1 .+-. 5 8/9 mg/kg, 2.3 i.v. (ns) once per week
[0261] 1.times.10.sup.6 PC-3M cells in 20 .mu.L of PBS are injected
into the left ventricle of the prostate of each male nude mouse
(n=9 per group). Treatments are started on day 14 after tumor cell
injection. epothilone B is administered i.v., once at 6 or 8 mg/kg,
or once per week at 4 or 5 mg/kg. Antitumor activity is expressed
as T/C% (mean tumor weight of treated animals divided by the mean
of tumor weight of control animals multiplied by 100). Differences
in body weights consider only animals surviving to the end of the
experiment (day 42). Statistical analyses on A body weight uses
paired t-tests comparing body weights before treatment to the end
of treatment; mice weigh.about.20-25 g at the start of treatment.
Not significant is indicated by the abbreviation ,,ns".
Example 7
Effect of Epothilone B Against Human Non-Small Cell Lung Carcinoma
A549
[0262] 3-10 million cells are implanted s.c. into the right
axillary (lateral) region of outbred athymic (nu/nu) mice, and are
allowed to grow until a tumor volume of approximately 100 mm.sup.3
is established. Epothilone B is formulated in 1% DMSO in 5% glucose
in distilled water (D5W), and administered i.v. either once only,
once per week, 3 times per week, or 5 times per week. Positive
controls are carried out with clinical formulations of TAXOL.RTM.
diluted 6 fold with D5W and administered i.v. 3.times./week.
[0263] Antitumor activity is expressed as % T/C (comparing .DELTA.
tumor volumes for treatment group to vehicle control group) at the
end of the experiment. Regressions are calculated using the
formula: -(T/T.sub.0-1).times.100% , where T is the tumor volume
for the treatment group at the end of the experiment, and T.sub.0
is the tumor volume at the beginning of the experiment. Statistical
significance is evaluated using a one-tailed Student's t-test.
[0264] Results: Table 7 summarizes results for A549 tumors.
Epothilone B induces significant tumor inhibition (T/C=41%), with
no detectable toxicity, when administered once at 6 mg/kg. A dose
of 4 mg/kg administered 1.times./week (4 mg/week) induces tumor
stasis (T/C=7%), but also produces a 13% body weight loss. By
comparison, at the dose of 1.5 mg/kg administered 3.times./week
(4.5 mg/week) all animals have to be euthanatized in the first week
of the experiment due to toxicity. Dosing with 0.5 mg/kg,
5.times./week (2.5 mg/week) induces tumor inhibition identical to
that of the once only regimen (T/C=41%), but apparent cumulative
toxicity results in a 23% body weight loss. TAXOL.RTM.,
administered 3.times./week at a dose of 20 mg/kg, does not inhibit
tumor growth, and induces a 16% body weight loss, with lethality in
1 of 8 mice.
7TABLE 7 Antitumor activity of Ep thilon B, compared to TAXOL
.RTM., on A549 non small c ll lung tumors in nude mice. Delta Mean
Delta % Dose Tumor Vol. % Regres- Body Dead/ Compound Regimen
(mg/kg) (mm.sup.3) % T/C sion Weight Total 1% DMSO/ 3x/week -- 439
-- none +8.9 0/7 D5W epothi- 5x/week 0.5 178 41** none -23.1 0/8
lone B epothi- 3x/week 1.5 NE NE NE NE 8/8 lone B epothi- 1x/weeks
4 32 7** none -13.2 0/8 lone B epothi- Once 6 178 41** none +8.5
0/8 lone B TAXOL .RTM. 3x/week 20 459 105 none -16.0 1/8 Cremo- --
-- 207 phor .RTM./etha- nol/D5W
[0265] Treatments are started on day 16 post implantation (10
million cells/animal). Epothilone B is administered i.v. once at 6
mg/kg (day 16), once weekly at 4 mg/kg (days 16, 23, and 30), three
times per week at 1.5 mg/kg (days 16, 18, 21, 23, 25, 28, 30, 32,
and 35), or five times per week at 0.5 mg/kg (days 16-18, 21-25,
28-32, and 35-36). TAXOL.RTM. is administered i.v. three times per
week at 20 mg/kg (days 16, 18, 21, 23, 25, 28, 30, 32, and 35) as
split doses of 10 mg/kg given one hour apart. Vehicle control (1%
DMSO/D5W) is administered i.v. three times per week (days 16, 18,
21, 23, 25, 28, 30, 32, and 35). All final data are recorded on day
37. A single asterisk (*) indicates p<0.05, and a double
asterisk (**) indicates p<0.01, using a one-tailed Student's
t-Test. "NE": not evaluable--animals euthanatized due to compound
toxicity.
Example 8
Antitumor Activity of Epothilone B Compared to TAXOL.RTM. on A549
Non-Small Cell Lung Tumors in Nude Mice
[0266] Materials and methods pertaining to the human tumor
xenograft model are as previously described. 10 or 1 million cells
(A549) are implanted s.c. into the right axillary (lateral) region
of outbred athymic (nu/nu) mice, and are allowed to grow until a
mass of approximately 100 mm.sup.3 is established. Epothilone B is
formulated in 1% DMSO in 5% glucose in distilled water (D5W), and
administered i.v. once weekly for 3 weeks. Positive controls are
carried out with clinical formulations of TAXOL.RTM. diluted 4 fold
with D5W and administered i.v. 3.times./week, for 3 weeks, in split
doses (2.times.10 mg/kg) given 1 hour apart.
[0267] Antitumor activity is expressed as % T/C (comparing A tumor
volumes for treatment group to vehicle control group) at the end of
the experiment. Regressions are calculated using the formula:
(T/T.sub.0-1).times.100%, where T is the tumor volume for the
treatment group at the end of the experiment, and T.sub.0 is the
tumor volume at the beginning of the experiment. Measurements are
taken for an additional 2 weeks after completion of the regular 3
weeks experiments, to evaluate reversibility of drug-induced body
weight loss, and sustainability of antitumor effects. Statistical
significance is evaluated using a one-tailed Student's t-test, and
Dunnett's, or Dunn's tests.
[0268] Results Table 8 summarizes results for A549 tumors, for the
standard 3 weeks experiment. Once weekly administration of
epothilone B produces statistically significant, dose dependent
inhibition of tumor growth, approaching tumor stasis at highest
concentrations of the drug. Epothilone B produces marked inhibition
of tumor growth at 3.5, and 3 mg/kg (T/C=15%, and 23%,
respectively). Both doses cause comparable, but reversible (see
Table 8) body weight loss of approximately 15%. The 2, and 1 mg/kg
doses produced 43%, and 74% T/C, that are statistically
significant, with no body weight gain at 2 mg/kg, and normal body
weight gain at 1 mg/kg. TAXOL.RTM., administered 3.times./week at a
split dose of 2.times.10 mg/kg, does not inhibit tumor growth, but
produces a 19% body weight loss.
8TABLE 8 Antitumor activity of epothilone B, compared to TAXOL
.RTM., n A549 non-small cell lung tumors in nude mice. Delta Mean
Delta % Dose Tumor Vol. Body Compound (mg/kg) (mm.sup.3 .+-. SEM) %
T/C Weight Dead/Total 1% DMSO/ -- 262 .+-. 26 -- +7.7 0/8 D5W
Epothilone B 1 195 .+-. 21 74*,** +10.8 0/8 Epothilone B 2 113 .+-.
21 43*,** +0.9 0/8 Epothilone B 3 60 .+-. 22 23*,** -16.4 0/8
Epothilone B 3.5 40 .+-. 15 15*,** -14.6 0/8 Cremophor .RTM./ --
207 .+-. 26 -- +8.3 0/8 ethanol/D5W TAXOL .RTM. 20 293 .+-. 56 142
-19.1 0/8
[0269] Treatments are started on day 13 post implantation (10
million cells/animal). Epothilone B is administered i.v. once
weekly for 3 weeks (days 13, 20, and 27) at doses of 1, 2, 3, and
3.5 mg/kg. TAXOL.RTM. is administered i.v. three times per week for
3 weeks at 20 mg/kg (days 14, 17, 19, 21, 24, 26, 28, 31, and 33)
as split doses of 10 mg/kg given one hour apart. Final data are
recorded on day 34. A single asterisk (*) indicates p<0.05 using
a one-tailed Student's t-Test, and a double asterisk (**) indicates
p<0.05 using a Dunnett's or Dunn's test.
[0270] Measurements are taken for an additional 2 weeks after
completion of the regular 3 weeks experiments, and the final data
for week 5 are summarized in Table 9. The antitumor effect remains
unchanged, while animal body weights have recovered. T/C values for
the 3.5, 3, 2, and 1 mg/kg dose levels of epothilone B are 12%,
16%, 49% and 72%, respectively, and all groups have normal weight
gain. TAXOL.RTM. remains ineffective, and animals show only a 2%
weight gain.
9TABLE 9 Antitumor activity of pothil ne B, compar d t TAXOL .RTM.,
on A549 non small cell lung tumors in nude mice (ext nded
bservation). Delta Mean Delta % Dose Tumor Vol. Body Compound
(mg/kg) (mm.sup.3 .+-. SEM) % T/C Weight Dead/Total 1% DMSO/ -- 472
.+-. 81 -- +11.8 0/8 D5W Epothilone B 1 339 .+-. 24 72 +12.6 0/8
Epothilone B 2 232 .+-. 39 49* +13.5 0/8 Epothilone B 3 75 .+-. 25
16*,** +13.2 0/8 Epothilone B 3.5 58 .+-. 25 12*,** +9.7 0/8
Cremophor .RTM./ -- 355 .+-. 80 -- +13.0 0/8 ethanol/D5W TAXOL
.RTM. 20 509 .+-. 123 144 +1.7 0/8
[0271] Measurements for the experiment described in Table 1 are
extended by additional two weeks. Final data are recorded on day
48. A single asterisk (*) indicates p<0.05 using a one-tailed
Student's t-Test, and a double asterisk (**) indicates p<0.05
using a Dunneff's or Dunn's test.
Example 9
Antitumor Effect of Epothilone B on ZR-75-1 Breast Tumors
[0272] Table 10 shows the results of an experiment where the effect
of TAXOL.RTM. and epothilone B on the breast cancer cell line
ZR-75-1 are compared. The methods that are used for this tumor
model have been described above.
[0273] It follows from the data that (judged on antitumor
efficiency) the best dosing schedule is one where 4 mg/kg are
administered weekly. However, mortality is observed at all dosages,
suggesting that the ZR-75-1 tumor may affect the overall health of
the mice in contrast to other tumor types.
10TABLE 10 Antitum r effect of epothilone B against subcutaneou ly
transplanted human e trogen-dependent ZR-75-1 br ast tumor in femal
BALB/c nude mice. Tumor Response Host Response Dose, .DELTA. Tumor
.DELTA. Body % Body Survival Com- Route, T/C Regres- Volume Weight
Weight (No. pound Schedule (%) sion (mm.sup.3) (g) Change
alive/total) Vehicle 25 ml/kg, 100 none 444 .+-. 58 0.9 4 .+-. 2
6/6 controls i.v. .+-. 0.4 every 7 days Epothi- 4 mg/kg, 46 none
208 .+-. 86 -1.6 -7 .+-. 5 4/6 lone B i.v. .+-. 1.1 every 14 days
Epothi- 4 mg/kg, 1 -8% 29 .+-. 16 -1.2 -5 .+-. 3 3/6 lone B iv.
(trans- .+-. 0.6 every 7 ient) days Epothi- 2 mg/kg, 40 none 227
.+-. 76 -3.5 -14 .+-. 6 5/6 lone B i.v. .+-. 1.6 every 7 days
Epothi- 1 mg/kg, 86 none 393 0.3 2 .+-. 5 4/6 lone B i.v. .+-. 135
.+-. 1.2 every 7 days
[0274] Tumor fragments of approximately 25 mg are implanted into
the left flank of each female nude mouse (n=6 per group); a
subcutaneous estrogen pellet is placed in the opposite flank.
Treatments are started on day 19 after tumor transplantation.
epothilone B is administered at 1, 2 or 4 mg/kg, i.v., either once
per week (days 19, 26 and 33) or every second week (days 19 and
33). Data presented are from animals surviving to day 47, the last
day of controls. Antitumor activity is expressed as T/C% (mean
increase of tumor volumes of treated animals divided by the mean
increase of tumor volumes of control animals multiplied by 100).
Tumor regression (%) represents the final mean tumor volume
compared to the mean tumor volume at the start of treatment.
Changes (A) in tumor volume represent the tumor volume on the last
treatment day minus the tumor volume on the first treatment
day.
Example 10
Antitumor Effect of Epothilone B in Comparison with 5-Fluorouracil
Against Subcuntaneously Transplanted Colo 205 Colon Tumors
[0275] Table 11 shows the ffect of epothilon B against
subcutaneously transplanted Colo 205 tumors, as well as the effect
of 5-fluorouracil. In contrast to the treatment of the HCT-15 cell
line tumors, where standard treatment with 5-fluorouracil or
treatment with TAXOL.RTM. is not effective, here the treatment with
5-fluorouracil is still effective, though much less so than with
epothilone B.
[0276] Together with the data from example 4 where the HCT-15 cells
do not respond to (are refractory to) both TAXOL.RTM. and the
standard colon cancer treatment with 5-fluorouracil, this shows
that epothilone B is indeed appropriate to treat tumors that are
refractory to known standard treatments. On the other hand, it is
also more effective where standard treatments work. A preferred
treatment schedule can be deduced which is 4 mg/kg every 2 weeks
(tumor regression, no dead animals). This treatment is even better
than that with 5-fluorouracil where no regression is found, but
only 4 out of 7 animals survive.
11TABLE 11 Antitumor effect of pothilone B in comparison with
5-fluor - uracil against subcutan ously transplant d human COLO 205
c lon tumors in f male BALB/c nude mic (day 32, four days post last
tr atm nt). Tumor Response Host Response Dose, .DELTA. Tumor % Body
Survival Com- Route, Regres- Volume .DELTA. Body Weight (No. pound
Schedule T/C sion (mm.sup.3) Weight (g) Change alive/total) Vehicle
25 ml/kg, 100% none 380 .+-. 96 2.7 .+-. 0.3 11 .+-. 2 7/7 controls
i.v. every 7 days Epothi- 4 mg/kg, Regres- -69% -62 .+-. 7 -1.1
.+-. 1.0 -4 .+-. 4 7/7 lone B i.v. sions every 14 days Epothi- 4
mg/kg, Regres- -87% -83 .+-. 7 -4.0 .+-. 0.2 -18 .+-. 1 5/7 lone B
i.v. sions every 7 days Epothi- 4 mg/kg, Regres- -66% -58 .+-. 11
2.0 .+-. 0.1 9 .+-. 1 5/7 lone B i.v. sions once 5-Fluo- 75 18 none
62 .+-. 12 2.2 .+-. 0.4 9 .+-. 2 4/7 rouracil mg/kg, i.v. every 7
days
[0277] Tumor fragments of approximately 25 mg are implanted into
the left flank of each female nude mouse (n=7 per group); a
subcutaneous estrogen pellet is placed on the opposite flank.
Treatments are started on day 14 after tumor transplantation.
epothilone B is administered at 4 mg/kg, i.v., either once or once
per week (days 14, 21, 28) or every second week (days 14 and 28).
5-fluorouracil is administered i.v. at 75 mg/kg on days 14, 21, 28.
Data presented are from animals surviving to day 32, four days
after the last treatments. Antitumor activity is expressed as T/C%
(mean increase of tumor volumes of treated animals divided by the
mean increase of tumor volumes of control animals multiplied by
100). Tumor regression (%) represents the final mean tumor volume
compared to the mean tumor volume at the start of treatment.
Changes (A) in tumor volumes represent the tumor volume on the last
treatment day minus the tumor volume on the first treatment
day.
Example 11
A phase 1, Dose-Finding Study of Single Agent Epothilone B
Administered Once Every Week to Adult Patients with Advanced Solid
Tumors
[0278] Number of centers 2
[0279] Objectives
[0280] Primary: To characterize the safety profile, including both
acute and cumulative toxicities, and determine the maximum
tolerated dose of single agent epothilone B administered by
intravenous infusion once every week to adult patients with
advanced solid tumors who have failed standard systemic therapy or
for whom standard systemic therapy does not exist.
[0281] Secondary: 1. To characterize the pharmacokinetics of single
agent epothilone B administered by intravenous infusion once every
week to this population of patients; data obtained are used in
concert with pharmacodynamic data (e.g. hematologic parameters), to
make pharmacokinetic/pharmacodynamic (PK/PD) correlations that help
predict safety and efficacy.
[0282] 2. To obtain preliminary evidence of antitumor activity of
single agent epothilone B administered by intravenous infusion once
every week to this population of patients.
[0283] 3. To correlate intratumor drug levels between adult
patients with advanced solid tumors receiving single agent
epothilone B by intravenous infusion once every week, to those
associated with efficacy in preclinical models.
[0284] 4. To gather pharmacogenetic information on tumors on tumor
biopsy samples where available and accessible pre- and post-therapy
in order to identify genes that correlate with efficacy and
response; this is performed either by genetic analysis of
individual gene expression (e.g. p53, Map4, and mdr1 expression
status) or by gene chip technology.
[0285] Design This is an open-label, dose-escalation study to
assess the safety, pharmacokinetics, and pharmacpdynamics of
epothilone B administered by intravenous infusion once every week
to adult patients with advanced solid tumors who have failed
standard systemic therapy or for whom standard systemic therapy
does not exist.
[0286] The treatment period consists of up to 24 weekly doses.
Patients experiencing unacceptable toxicity or disease progression
are discontinued prematurely. Patients achieving a complete or
partial response, or patients with stable disease at th end of 24
doses continue further treatment according to an extension protocol
at the discretion of the investigator and after approval by the
sponsor. Eligible patients receive additional cycles until disease
progression or unacceptable toxicity.
[0287] The standard Phase 1 protocol design of enrolling 3-6
patients per cohort to establish the MTD is employed. Dose
escalation proceed according to a modified Fibonacci scheme and is
based on toxicities from the first 4 weekly doses for each cohort
of patients. The starting dose is 0.1 mg/m.sup.2, with subsequent
doses as follows: 0.2, 0.3, 0.5, 0.7, and 0.9 mg/m.sup.2.
[0288] The provisional MTD are defined as the dose level
immediately below that at which dose limiting toxicity (DLT) is
observed in at least two out of 3-6 patients. The cohort defined as
the provisional MTD then enrolls additional patients to a total of
12 to confirm the MTD through further evaluation of the safety,
pharmacokinetic, and pharmacodynamic profiles of epothilone B.
[0289] All toxicities are defined according to the revised US
National Cancer Institute Common Toxicity Criteria. DLTs are
defined in the protocol; in general, however, the nature of a DLT
is such that it is considered unacceptable even in the setting of
an incurable solid tumor.
[0290] Patients
[0291] Inclusion Criteria
[0292] The following criteria are to be met for inclusion into the
study:
[0293] 1. Male or female patients .gtoreq.8 years of age.
[0294] 2. Histologically documented advanced solid tumor, who have
failed standard systemic therapy and up to 1 additional systemic
therapy, or for whom standard systemic therapy does not exist.
[0295] 3. At least one measurable, evaluable, or non-evaluable site
of disease as defined by Southwestern Oncology Group (SWOG) Solid
Tumor Response Criteria including tumor marker value that is above
the institutional upper limit of normal.
[0296] 4. Women of childbearing potential must have a negative
serum .beta.-HCG pregnancy test prior to the initiation of study
drug. Male and female patients of reproductive potential must agree
to employ an effective method of birth control throughout the study
and for up to 3 months following discontinuation of study drug.
[0297] 5. World Health Organization (WHO) Performance Status Score
of <2.
[0298] 6. Life expectancy of at last 3 months.
[0299] 7. Written informed consent obtained prior to any screening
procedures.
[0300] Exclusion Criteria
[0301] Exclusion from the study is required if any of the following
apply:
[0302] 1. Female patients who are pregnant or breast-feeding.
Postmenopausal women must be amenorrheic for at least 12 months to
be considered of non-childbearing potential.
[0303] 2. Patient has a severe and/or uncontrolled medical disease
(i.e., uncontrolled diabetes, congestive heart failure, myocardial
infarction within 6 months of study, chronic renal disease, or
active uncontrolled infection).
[0304] 3. Patient has a known brain metastasis.
[0305] 4. Patient has an acute or known chronic liver disease
(i.e., chronic active hepatitis, cirrhosis).
[0306] 5. Patient has a known diagnosis of human immunodeficiency
virus (HIV) infection.
[0307] 6. Patient has received any investigational agent within 30
days prior to study entry.
[0308] 7. Patient received chemotherapy within 4 weeks (6 weeks for
nitrosoureas or mitomycin C) prior to study entry.
[0309] 8. Patient received prior radiation therapy within 4 weeks
prior to study entry.
[0310] 9. Patient previously received radiotherapy to .gtoreq.25%
of the bone marrow.
[0311] 10. Patient had a major surgery within 2 weeks prior to
study entry.
[0312] 11. Patient has a history of non-compliance to medical
regimens.
[0313] 12. Patient has impairment of hepatic, renal or hematologic
function as defined by the following laboratory parameters:
[0314] Platelet count <100.times.10.sup.9/L
[0315] Absolute neutrophil count (ANC)<1.5.times.10.sup.9/L
[0316] Serum ALT (SGPT)>2.5.times.institutional upper limit of
normal (IULN)
[0317] Serum total bilirubin>1.5.times.IULN
[0318] Serum creatinine>1.5.times.IULN
[0319] 14. Patient is <5 years free of another primary
malignancy or, in the case of non-melanomatous skin cancer and
cervical carcinoma in situ, has active disease.
[0320] Sample Size This study requires about 40 patients.
[0321] Treatments Epothilone B is supplied in individual 2 ml glass
vials formulated as 1 mg/l ml of the clear, colorless intravenous
concentrate. The substance is formulated in polyethylene glycol 300
(PEG 300) and diluted with 50 to 100 ml 0.9% Sodium Chloride
Injection, USP, to achieve the desired final concentration of the
drug for infusion. It is administered as a single 30-minute
intravenous infusion every 7 days.
[0322] The starting dose level is 0.1 mg/M.sup.2. This dose is
calculated as one-third of the toxic dose low (TDL) in the most
sensitive species studied which, for epothilone B, is the dog. As
described above, dose escalation proceeds according to a modified
Fibonacci scheme. The study defines treatment delays, dose
reductions, or withdrawal from treatment for individuals
experiencing hematologic or other toxicities known to result from
epothilone B. Treatment continues to a maximum of 24 weekly doses
unless the patient experiences disease progression or unacceptable
toxicity. At the end of 24 doses, patients who have achieved a
complete or partial response and patients who have had stable
disease may continue further treatment according to an extension
protocol at the discretion of the investigator and after approval
by the sponsor.
[0323] Safety
[0324] Variables The safety of epothilone B is assessed by physical
examination and evaluation of vital signs, clinical laboratory
results, adverse events, and use of concomitant medications.
Adverse events are both elicited and volunteered and are graded
using the revised US National Cancer Institute Expanded Common
Toxicity Criteria.
[0325] Efficacy
[0326] Variables Although this phase 1 study is not designed to
detect efficacy, activity is demonstrated as a function of the rate
of objective tumor response and length of progression-free and
overall survival. Baseline tumor evaluations include optimal
assessment of all measurable, evaluable, and nonevaluable disease.
Evaluations include physical examination and chest roentgenogram
and, as appropriate, computerized tomogram of the thorax, abdomen
and pelvis; sonogram of the abdomen and pelvis; bone scintigram,
with bone roentgenogram of all known osseous lesions; and
determination of tumor marker values. Follow-up studies are
obtained every 6 weeks and after cessation of treatment.
[0327] Objective status is clinically evaluated using the Novartis
guidelines, which are based on the SWOG response criteria. All
complete and partial responses must be confirmed by a second
assessment at least four weeks later. Best tumor response are
calculated for each patient using the SWOG respons criteria.
[0328] Pharmacokinetics The following pharmacokinetic parameters
are calculated and analyzed for cycles 1 and 2: t.sub.max,
C.sub.max, .lambda..sub.Z, t.sub.1/2, AUC, and RA. RA=the ratio of
AUC.sub..tau.cycle2/AUC.sub..tau.cycle1 is evaluated as an index of
accumulation. Preliminary assessment of dose proportionality is
based on AUC from the last dose among different dose groups. PK/PD
correlations with observed toxicities (e.g., hematopoietic) are
performed as a predictor of safety.
[0329] Pharmacodynamics Tumor biopsy samples are obtained where
feasible and accessible, pre-therapy and after the first cycle of
therapy. These biopsy samples are prepared for analysis of their
gene expression using gene chip technology, then separately
analyzed for p53 status, MAP4 RNA expression, and mdr1 RNA
expression.
[0330] Statistical
[0331] methods Patients with treatment-emergent clinical adverse
events (especially those with dose-limiting toxicity) or with
laboratory, vital sign, or physical examination abnormalities
(newly occurring or worsening from baseline) are identified and the
values are flagged. The rate of abnormalities is tabulated by
cohort. Objective response rates (including both complete and
partial responses) are presented by cohort. Descriptive statistics
are used to summarize the basic pharmacokinetic parameters by
cohort.
Example 12
A Phase 1, Dose-Finding Study of Single Agent EPO906 (Epothilone B)
Administered Once Every Three Weeks to Adult Patients with Advanced
Solid Tumors
[0332] No. of centers: 2
[0333] Locations Glasgow, UK, & Newcastle, UK
[0334] Objectives
[0335] Primary: To characterize the safety profile, including both
acute and cumulative toxicities, and determine the maximum
tolerated dose of single agent epothilone B administered by
intravenous infusion once every three weeks to adult patients with
advanced solid tumors who have failed standard systemic therapy or
for whom standard systemic therapy does not exist
[0336] Secondary: 1. To characterize the pharmacokinetics of single
agent epothilone B administered by intravenous infusion once every
three weeks to this population of patients; data obtained are used
in concert with pharmacodynamic data to make
pharmacokinetic/pharmacodynamic (PK/PD) correlations that help
predict safety and efficacy
[0337] 2. To obtain preliminary evidence of antitumor activity of
single agent epothilone B administered by intravenous infusion once
every three weeks to this population of patients
[0338] 3. To correlate intratumor drug levels between adult
patients with advanced solid tumors receiving single agent
epothilone B by intravenous infusion once every three weeks to
those associated with efficacy in preclinical models
[0339] 4. To gather information on tumors from tumor biopsy samples
where available and accessible pre- and post-therapy in order to
identify biological factors that correlate with efficacy and
response
[0340] Design This is an open-label, dose-escalation study to
assess the safety, pharmacokinetics, and pharmacodynamics of
epothilone B administered by intravenous infusion once every three
weeks to adult patients with advanced solid tumors who have failed
standard systemic therapy or for whom standard systemic therapy
does not exist.
[0341] The treatment period consists of up to six 21-day cycles.
Patients experiencing unacceptable toxicity or disease progression
are discontinued prematurely. Patients achieving a complete or
partial response, or patients with stable disease at the end of six
cycles continue further treatment according to an extension
protocol at the discretion of the investigator and after approval
by the sponsor. Eligible patients receive additional cycles until
disease progression or unacceptable toxicity are observed.
[0342] In the absence of dose-limiting toxicity (DLT), dose
escalation proceeds as follows:
[0343] 1. First dos escalation: 100% dos increase (unless grade 2
toxicity is identified in first cohort, in which case dose
escalation is 25%-67%)
[0344] 2. Dose escalations following 100% dose increase from first
to second cohort: 67% dose increases until grade 2 toxicity is
identified
[0345] 3. Final dose escalations following identification of grade
2 toxicity: 25%-67% dose increases, based on consensus reached
among the investigators and the sponsor
[0346] Dose escalation is based on toxicities from the first cycle
for each cohort of patients. The provisional maximum tolerated dose
(MTD) is defined as the dose level immediately below that at which
DLT is observed in at least two out of 3-6 patients. The cohort
defined as the provisional MTD then enrolls additional patients to
a total of 12 to confirm the MTD through further evaluation of the
safety, pharmacokinetic, and pharmacodynamic profiles of epothilone
B.
[0347] Intrapatient dose escalation will not be permitted.
[0348] All toxicities are defined according to the revised US
National Cancer Institute Common Toxicity Criteria. DLTs are
defined in the protocol; in general, however, the nature of a DLT
is such that it is considered unacceptable even in the setting of
an incurable solid tumor.
[0349] Patients
[0350] Inclusion Criteria
[0351] The following criteria must be met for inclusion into the
study:
[0352] 1. Male or female patients .gtoreq.18 years of age.
[0353] 2. Histologically documented advanced solid tumor, who have
failed standard systemic therapy and up to 1 additional systemic
therapy, or for whom standard systemic therapy does not exist.
[0354] 3. At least one measurable, evaluable, or non-evaluable site
of disease as defined by Southwestern Oncology Group (SWOG) Solid
Tumor Response Criteria including tumor marker value that is above
the institutional upper limit of normal.
[0355] 4. Women of childbearing potential must have a negative
serum .beta.-HCG pr gnancy test prior to th initiation of study
drug. Male and female patients of reproductive potential must agree
to employ an effective method of birth control throughout the study
and for up to 3 months following discontinuation of study drug.
[0356] 5. World Health Organization (WHO) Performance Status Score
of .ltoreq.2.
[0357] 6. Life expectancy of at least 3 months.
[0358] 7. Written informed consent is obtained prior to any
screening procedures.
[0359] Exclusion Criteria
[0360] Exclusion from the study is required if any of the following
apply:
[0361] 1. Female patients who are pregnant or breast-feeding.
Postmenopausal women must be amenorrheic for at least 12 months to
be considered of non-childbearing potential.
[0362] 2. Patient has a severe and/or uncontrolled medical disease
(i.e., uncontrolled diabetes, congestive heart failure, myocardial
infarction within 6 months of study, chronic renal disease, or
active uncontrolled infection).
[0363] 3. Patient has a known brain metastasis.
[0364] 4. Patient has an acute or known chronic liver disease
(i.e., chronic active hepatitis, cirrhosis).
[0365] 5. Patient has a known diagnosis of human immunodeficiency
virus (HIV) infection.
[0366] 6. Patient has received any investigational agent within 30
days prior to study entry.
[0367] 7. Patient received chemotherapy within 4 weeks (6 weeks for
nitrosoureas or mitomycin C) prior to study entry.
[0368] 8. Patient received prior radiation therapy within 4 weeks
prior to study entry.
[0369] 9. Patient previously received radiotherapy to .gtoreq.25%
of the bone marrow.
[0370] 10. Patient had a major surgery within 2 weeks prior to
study entry.
[0371] 11. Patient has a history of non-compliance to medical
regimens.
[0372] 12. Patient has impairment of hepatic, renal or hematologic
function as defined by the following laboratory parameters:
[0373] Platelet count <100.times.10.sup.9/L
[0374] Absolute neutrophil count (ANC)<1.5.times.10.sup.9/L
[0375] Serum ALT (SGPT) or AST (SGOT)>2.5.times.institutional
upper limit of
[0376] normal (IULN) (>5.times.IULN for patients with hepatic
metastases)
[0377] Serum total bilirubin>1.5.times.IULN
[0378] Serum creatinine>1.5.times.IULN
[0379] 13. Patient is <5 years free of another primary
malignancy; however, nonmelanomatous skin cancer and cervical
carcinoma in situ are excluded only if the patient has active
disease.
[0380] Sample size This study requires about 40 patients.
[0381] Treatments epothilone B is supplied in individual 2 ml glass
vials formulated as 1 mg/l ml of the clear, colorless intravenous
concentrate. The substance is formulated in polyethylene glycol 300
(PEG 300) and diluted with 50 or 100 ml 0.9% Sodium Chloride
Injection, USP, to achieve the desired final concentration of the
drug for infusion. It is administered as a single 30-minute
intravenous infusion every 21 days for six cycles.
[0382] The starting dose level is 0.3 mg/m.sup.2. This dose is
calculated as one-third of the toxic dose low (TDL) in the most
sensitive species studied which, for epothilone B, is the dog.
Since there are no mortalities at the lower of the 2 doses
administered to dogs in the GLP toxicology study--0.1 mg/kg,
repeated once 3 weeks later--the TDL is estimated to be in the
range of 0.5 mg/kg. Using a factor of 20 to convert mg/kg in the
dog to mg/m.sup.2 in humans, this starting dose is calculated
as:
{fraction (1/3 )}.times.0.05 mg/kg.times.20 kg/m.sup.2=0.3
mg/m.sup.2
[0383] Dose escalation proceeds according to the scheme outlined
above.
[0384] The study defines treatment delays, dose reductions, or
withdrawal from treatment for individuals experiencing hematologic
or other toxicities known to result from epothilone B. Treatment
continues to a maximum of 6 cycles unless the patient exp riences
disease progr ssion or unacceptable toxicity.
[0385] At the end of 6 cycles, patients who have achieved a
complete or partial response and patients who have had stable
disease may continue further treatment according to an extension
protocol at the discr tion of the investigator and after approval
by the sponsor.
[0386] Safety
[0387] variables The safety of epothilone Bis assessed by physical
examination and evaluation of vital signs, clinical laboratory
results, adverse events, and use of concomitant medications.
Adverse events are both elicited and volunteered and are graded
using the revised US National Cancer Institute Common Toxicity
Criteria.
[0388] Efficacy
[0389] variables Although this phase 1 study is not designed to
detect efficacy, activity is demonstrated as a function of the rate
of objective tumor response and length of progression-free and
overall survival. Baseline tumor evaluations include optimal
assessment of all measurable, evaluable, and nonevaluable disease.
Evaluations include physical examination and chest roentgenogram
and, as appropriate, computerized tomogram of the thorax, abdomen
and pelvis; sonogram of the abdomen and pelvis; bone scintigram,
with bone roentgenogram of all known osseous lesions; and
determination of tumor marker values. Follow-up studies are
obtained every two cycles and after cessation of treatment.
[0390] Objective status is clinically evaluated using the Novartis
guidelines, which are based on the SWOG response criteria. All
complete and partial responses must be confirmed by a second
assessment at least four weeks later.
[0391] Best tumor response are calculated for each patient using
the SWOG response criteria.
[0392] Pharmacokinetics The following pharmacokinetic parameters
are calculated and analyzed for cycles 1 and 2: t.sub.max,
C.sub.max, .lambda..sub.z, t.sub.1/2, AUC, and R.sub.A. R.sub.A=the
ratio of AUC.sub..tau.cycle2/AUC.sub..tau.cycle1 is evaluated as an
index of accumulation. Preliminary assessment of dose
proportionality is based on AUC from the last dose among different
dose groups.
[0393] PK/PD correlations with observed toxicities (e.g.,
hematopoietic) are performed as a predictor of safety.
[0394] Pharmacodynamics Tumor biopsy samples are obtained where
feasible and accessible pretherapy and after the first cycle of
therapy in order to identify biological factors that correlate with
efficacy and response.
[0395] Statistical
[0396] methods Patients with treatment-emergent clinical adverse
events (especially those with dose-limiting toxicity) or with
laboratory, vital sign, or physical examination abnormalities
(newly occurring or worsening from baseline) are identified and the
values are flagged. The rate of abnormalities is tabulated by
cohort. Objective response rates (including both complete and
partial responses) are presented by cohort. Descriptive statistics
are used to summarize the basic pharmacokinetic parameters by
cohort.
[0397] Discussion: Taken together, the examples provide evidence
that treatment with epothilone B is effective
[0398] a) also against a tumor where standard treatment fails, e.g.
in colon tumor where 5-fluorouracil treatment fails, or where
TAXOL.RTM. treatment fails;
[0399] b) also against a tumor where TAXOL.RTM. treatment fails,
e.g. lung, especially non-small cell lung cancer, and/or
epidermoid, especially cervical, tumors;
[0400] c) also against orthotopic tumors and the formation of
metastases, e.g. in prostate tumors;
[0401] d) also against breast cancer where in in vitro assays
(example 3) epothilone B shows higher activity than TAXOL.RTM..
[0402] The preferred dosage regimens center around an area of
weekly treatment with about {fraction (1/3 )} to {fraction (2/3 )}
of the MTD up to treatment once with a dose up to the MTD, with a
kind of best treatment area lying at the weekly up to three-weekly
administration.
* * * * *