U.S. patent application number 13/648597 was filed with the patent office on 2013-04-25 for herbal composition phy906 and its use in chemotherapy.
This patent application is currently assigned to YALE UNIVERSITY. The applicant listed for this patent is YALE UNIVERSITY. Invention is credited to Yung-Chi CHENG, Zaoli JIANG, Shwu-Huey LIU.
Application Number | 20130101688 13/648597 |
Document ID | / |
Family ID | 39690520 |
Filed Date | 2013-04-25 |
United States Patent
Application |
20130101688 |
Kind Code |
A1 |
LIU; Shwu-Huey ; et
al. |
April 25, 2013 |
HERBAL COMPOSITION PHY906 AND ITS USE IN CHEMOTHERAPY
Abstract
This invention provides herbal compositions useful for
increasing the therapeutic index of chemotherapeutic compounds.
This invention also provides methods useful for improving the
quality of life of an individual undergoing chemotherapy.
Furthermore, this invention improves the treatment of disease by
increasing the therapeutic index of chemotherapy drugs by
administering the herbal composition PHY906 to a mammal undergoing
such chemotherapy.
Inventors: |
LIU; Shwu-Huey; (Madison,
CT) ; JIANG; Zaoli; (Woodbridge, CT) ; CHENG;
Yung-Chi; (Woodbridge, CT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
YALE UNIVERSITY; |
New Haven |
CT |
US |
|
|
Assignee: |
YALE UNIVERSITY
New Haven
CT
|
Family ID: |
39690520 |
Appl. No.: |
13/648597 |
Filed: |
October 10, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12527302 |
Jan 29, 2010 |
8309141 |
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PCT/US08/53965 |
Feb 14, 2008 |
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13648597 |
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11100433 |
Apr 7, 2005 |
7534455 |
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12527302 |
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10220876 |
Dec 30, 2002 |
7025993 |
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PCT/US01/07353 |
Mar 8, 2001 |
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11100433 |
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09522055 |
Mar 9, 2000 |
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10220876 |
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60901310 |
Feb 15, 2007 |
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60625943 |
Nov 9, 2004 |
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Current U.S.
Class: |
424/725 |
Current CPC
Class: |
A61K 31/7068 20130101;
A61K 36/484 20130101; A61K 36/725 20130101; A61P 35/00 20180101;
A61K 36/539 20130101; A61K 31/4412 20130101; A61K 36/65 20130101;
A61K 36/484 20130101; A61K 2300/00 20130101; A61K 36/539 20130101;
A61K 2300/00 20130101; A61K 36/65 20130101; A61K 2300/00 20130101;
A61K 36/725 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/725 |
International
Class: |
A61K 36/65 20060101
A61K036/65 |
Claims
1-17. (canceled)
18. A method of treating cancer in a mammal comprising
co-administering a therapeutically effective amount of i) an herbal
preparation consisting essentially of Scutellaria baicalensis,
Glycyrrhiza uralensis, Ziziphus jujuba and Paeonia lactiflora; and
ii) a chemotherapeutic formulation comprising a chemotherapeutic
compound.
19. The method of claim 18, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via oral or parenteral route.
20. The method of claim 19, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via intravenous route.
21. The method of claim 18, wherein the herbal preparation consists
of Scutellaria baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba
and Paeonia lactiflora, and one or more pharmaceutically acceptable
carriers.
22. The method of claim 18, wherein the herbal preparation is
administered before the administration of the chemotherapeutic
formulation.
23. The method of claim 18, wherein the herbal preparation is
administered after the administration of the chemotherapeutic
formulation.
24. The method of claim 18, wherein the herbal preparation is
administered concurrently with the administration of the
chemotherapeutic formulation.
25. A method of relieving side effects of a chemotherapeutic
compound in a mammal comprising co-administering a therapeutically
effective amount of i) an herbal preparation consisting essentially
of Scutellaria baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba
and Paeonia lactiflora; and ii) a chemotherapeutic formulation
comprising a chemotherapeutic compound.
26. The method of claim 25, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via oral or parenteral route.
27. The method of claim 26, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via intravenous route.
28. The method of claim 25, wherein the herbal preparation consists
of Scutellaria baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba
and Paeonia lactiflora, and one or more pharmaceutically acceptable
carriers.
29. The method of claim 25, wherein the herbal preparation is
administered before the administration of the chemotherapeutic
formulation.
30. The method of claim 25, wherein the herbal preparation is
administered after the administration of the chemotherapeutic
formulation.
31. The method of claim 25, wherein the herbal preparation is
administered concurrently with the administration of the
chemotherapeutic formulation.
32. A method of enhancing anticancer activity of a chemotherapeutic
compound in a mammal comprising administering a therapeutically
effective amount of i) an herbal preparation consisting essentially
of Scutellaria baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba
and Paeonia lactiflora; and ii) a chemotherapeutic formulation
comprising a chemotherapeutic compound.
33. The method of claim 32, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via oral or parenteral route.
34. The method of claim 33, wherein the herbal preparation is
administered via oral route, and the chemotherapeutic formulation
is administered via intravenous route.
35. The method of claim 32, wherein the herbal preparation consists
of Scutellaria baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba
and Paeonia lactiflora, and one or more pharmaceutically acceptable
carriers.
36. The method of claim 32, wherein the herbal preparation is
administered before the administration of the chemotherapeutic
formulation.
37. The method of claim 32, wherein the herbal preparation is
administered after the administration of the chemotherapeutic
formulation.
38. The method of claim 32, wherein the herbal preparation is
administered concurrently with the administration of the
chemotherapeutic formulation.
39. A method of treating cancer in a mammal comprising separately
administering a therapeutically effective amount of i) an herbal
preparation consisting essentially of Scutellaria baicalensis,
Glycyrrhiza uralensis, Ziziphus jujuba and Paeonia lactiflora; and
ii) a chemotherapeutic formulation comprising a chemotherapeutic
compound.
40. A method of relieving side effects of a chemotherapeutic
compound in a mammal comprising separately administering a
therapeutically effective amount of i) an herbal preparation
consisting essentially of Scutellaria baicalensis, Glycyrrhiza
uralensis, Ziziphus jujuba and Paeonia lactiflora; and ii) a
chemotherapeutic formulation comprising a chemotherapeutic
compound.
41. A method of enhancing anticancer activity of a chemotherapeutic
compound in a mammal comprising separately administering a
therapeutically effective amount of i) an herbal preparation
consisting essentially of Scutellaria baicalensis, Glycyrrhiza
uralensis, Ziziphus jujuba and Paeonia lactiflora; and ii) a
chemotherapeutic formulation comprising a chemotherapeutic
compound.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of pending U.S. patent
application Ser. No. 12/527,302, which is the U.S. national phase
of International Patent Application No. PCT/US2008/053965, filed on
Feb. 14, 2008, which claims the benefit of U.S. Provisional Patent
Application Ser. No. 60/901,310, filed on Feb. 15, 2007, and which
also claims the benefit under 35 U.S.C. .sctn.120 as a
continuation-in-part of U.S. patent application Ser. No.
11/100,433, filed Apr. 7, 2005, now U.S. Pat. No. 7,534,455, which
claims the benefit of U.S. Provisional Patent Application Ser. No.
60/625,943, filed Nov. 9, 2004, and which in turn is a
continuation-in-part of U.S. patent application Ser. No.
10/220,876, now U.S. Pat. No. 7,025,993, which is the U.S. national
phase of International Patent Application No. PCT/US2001/007353,
filed Mar. 8, 2001, which in turn is a continuation-in-part of U.S.
patent application Ser. No. 09/522,055, filed Mar. 9, 2000 and now
abandoned. The contents of the aforementioned applications are
herein incorporated by reference in their entirety for all
purposes. The content of International Patent Application No.
PCT/US2005/040605, filed Nov. 9, 2005, is also herein incorporated
herein by reference in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to herbal compositions and the
use of them for enhancing the therapeutic effects of
chemotherapeutic compounds.
BACKGROUND OF THE INVENTION
[0003] Cancer remains one of the major causes of death around the
world. Specifically, cancer is the second overall cause of death in
the United States. Gastrointestinal cancers, including colorectal,
liver, and pancreatic cancers, are of particular concerns not only
because of their high incidence rates, but also because of their
high mortality rate, especially in pancreatic and liver cancer
patients (1-4). From years 1992-1999, studies revealed that the
five-year relative survival rate of colorectal cancer was 62.3%
while that of liver cancer was 6.9% and 4.4% for pancreatic cancer.
The median survival of liver cancer was 3.5 weeks to 6 months while
it was 4 to 6 months for pancreatic cancer (3). With only very poor
chemotherapeutic regimens available, pancreatic cancer has the
highest mortality rate among all cancers in the United States, with
a less than 5% survival rate 5 years from diagnosis (3). Although
several regimens are currently used in clinical trials for
hepatocellular carcinoma, there is no FDA-approved chemotherapeutic
agent available. The low survival rates for both pancreatic and
hepatocellular cancers are attributed to many factors, including
that diagnosis is difficult, the tumor growth is highly aggressive,
surgical removal of tumor is of low probability, and the tumor has
a high rate of chemotherapy resistance.
BRIEF DESCRIPTION OF THE DRAWINGS
[0004] FIG. 1 shows the effect of PHY906 (500 mg/kg, bid, D1-4 and
8-11) on tumor growth in Sorafenib (30 mg/kg, po, bid,
D1-14)-treated BDF-1 mouse bearing mouse colon 38 tumors. Sorafenib
(30 mg/kg) was given orally twice a day for a consecutive 14 days.
PHY906 (500 mg/kg) was given orally 30 min before sorafenib twice a
day on days 1-4 and days 8-11 (N=5 in each group).
[0005] FIG. 2 shows the effect of PHY906 (500 mg/kg, bid, D1-4,
8-11 and 15-18) on tumor growth in Sorafenib (30 mg/kg, po, bid,
D1-20)-treated nude mice bearing human HepG2 tumors. Sorafenib (30
mg/kg) was given orally twice a day for a consecutive 20 days.
PHY906 (500 mg/kg) was given orally 30 min before sorafenib twice a
day on days 1-4, 8-11 and 15-18 (N=5 in each group).
[0006] FIG. 3 shows the impact of PHY906 and Sorafenib on blood
vessels from the liver of NCr-nude mice bearing human HepG2
xenografts. Tissue sections were prepared from formalin-fixed,
paraffin-embedded liver cancer specimens. Immunohistochemical
staining was done using specific antibodies against CD31 (brown)
and nuclear DNA (blue).
[0007] FIG. 4 shows impact of PHY906 and Sorafenib on VEGF level
from the liver of NCr-nude mice bearing human HepG2 xenografts.
Tissue sections were prepared from formalin-fixed,
paraffin-embedded liver cancer specimens. Immunohistochemical
staining was done using specific antibodies against VEGF (brown)
and nuclear DNA (blue).
[0008] FIG. 5 shows the impact of PHY906 and Sorafenib on
HIF-1.alpha. level from the liver of NCr-nude mice bearing human
hepG2 xenografts. Tissue sections were prepared from
formalin-fixed, paraffin-embedded liver cancer specimens.
Immunohistochemical staining was done using specific antibodies
against HIF-1.alpha.(brown) and nuclear DNA (blue).
[0009] FIG. 6 shows the effect of PHY906 on tumor growth in
Capecitabine-treated NCr-nude mice bearing human Panc-1 tumor.
Capecitabine (720 mg/kg) was given orally twice a day on days 1-7,
15-21 and 29-32. PHY906 was given orally 30 min before capecitabine
twice a day on days 1-4, 8-11, 15-18, 22-25 and 29-32 at 500 mg/kg
(N=5 in each group).
[0010] FIG. 7. Effect of PHY906 on Body Weight in FU/LV Treated
BDF-1 Mice Bearing Colon 38 Tumor. Sequential administration of LV
(100 mg/kg) and FU (100 mg/kg) was given intraperitoneally during 1
hr period on day 0 only, as described in Materials and Methods.
PHY906 was given orally 30 min after initial dose of LV on day 0
and continued twice a day for 4-days at 500 mg/kg (N=5 in each
group).
[0011] FIG. 8. Effect of PHY906 on Tumor Growth in FU/LV Treated
BDF-1 Mice Bearing Colon 38 Tumor. Sequential administration of LV
(100 mg/kg) and FU (100 mg/kg) was given intraperitoneally during 1
hr period on day 0 only, as described in Materials and Methods.
PHY906 was given orally 30 min after initial dose of LV on day 0
and continued twice a day for 4 days at 500 mg/kg (N=5 in each
group).
[0012] FIG. 9. Effect of PHY906 on Hematological Change in FU/LV
Treated BDF-1 Mice Bearing Colon 38 Tumor. Sequential
administration of LV (100 mg/kg) and FU (100 mg/kg) was given
intraperitoneally during 1 hr period on day 0 only, as described in
Materials and Methods. PHY906 was given orally 30 min after initial
dose of LV on day 0 and continued twice a day for 4-days at 500
mg/kg (N=5 in each group).
[0013] FIG. 10 A-C. Pharmacokinetics of CPT-11/FU/LV in Plasma.
PHY906-6 is the clinical batch of PHY906. SN-38 is an active
metabolite of CPT-11, FUR+FUMP are nucleoside and nucleotide
metabolites of FU.
[0014] FIG. 11A-C. Pharmacokinetics of CPT-11/FU/LV in Liver.
PHY906-6 is the clinical batch of PHY906. SN-38 is an active
metabolite of CPT-11. FUR+FUMP are nucleoside and nucleotide
metabolites of FU.
[0015] FIG. 12 A-B. Pharmacokinetics of CPT-11/FU/LV in Tumor.
PHY906-6 is the clinical batch of PHY906. SN-38 is an active
metabolite of CPT-11.
[0016] FIG. 13. Effect on Different Dosage of CPT-11 in Non-Tumor
Bearing BDF-1 Mice. CPT-11 was given intraperitoneally (i.p.) on
day 0 only (N=5 in each group).
[0017] FIG. 14. Effect of PHY906 on Body Weight in CPT-11 Treated
BDF-1 Mice Bearing Colon 38 Tumor. CPT-11 (400 mg/kg) was given
intraperitoneally on day 0 only. PHY906 was given orally twice a
day for 8 days beginning on day 0 at the dose indicated (N=5 in
each group).
[0018] FIG. 15. Effect of PHY906 on Tumor Growth in CPT-11 Treated
BDF-1 Mice Bearing Colon 38 Tumor. CPT-11 (400 mg/kg) was given
intraperitoneally on day 0 only. PHY906 (500 mg/kg) was given
orally twice a day for 8 days beginning on day 0. The p values were
calculated using the Student's paired t-test. (N=5 in each
group)
[0019] FIG. 16. Effect of PHY906 on Hematological Change in CPT-11
Treated BDF-1 Mice Bearing Colon 38 Tumor (N=5 in each group).
CPT-11 (400 mg/kg) was given intraperitoneally on day 0 only.
PHY906 (500 mg/kg) was given orally twice a day for 4 days
beginning on day 0 (N=5 in each group).
[0020] FIG. 17. Effect of PHY906 on Tumor Growth in CPT-11/FU/LV
Treated BDF-1 Mice Bearing Colon 38 Tumor. Sequential
administration of LV (100 mg/kg), CPT-11 (200 mg/kg) and FU (100
mg/kg) was given intraperitoneally during 1 hr period on day 0
only, as described in Materials and Methods. PHY906 was given
orally 30 min after initial dose of LV on day 0 and continued twice
a day for 4 days at 500 mg/kg (N=5 in each group).
[0021] FIG. 18. Effect of PHY906 on Tumor Growth in CPT-11/FU/LV
Treated BDF-1 Mice Bearing Colon 38 Tumor. Sequential
administration of LV (100 mg/kg), CPT-11 (300 mg/kg) and FU (100
mg/kg) was given intraperitoneally during 1 hr period on day 0
only, as described in Materials and Methods. PHY906 was given
orally 30 min after initial dose of LV on day 0 and continued twice
a day for 4 days at 500 mg/kg (N=5 in each group).
[0022] FIG. 19. Effect of PHY906 on Body Weight Change in
CPT-11/FU/LV Treated BDF-1 Mice Bearing Colon 38 Tumor. Sequential
administration of LV (100 mg/kg), CPT-11 (300 mg/kg) and FU (100
mg/kg) was given intraperitoneally during 1 hr period on day 0
only, as described in Materials and Methods. PHY906 was given
orally 30 min after initial dose of LV on day 0 and continued twice
a day for 4 days at 500 mg/kg (N=5 in each group).
[0023] FIG. 20. Effect of PHY906 on the Tumor Growth in CPT-11
Treated NCr-Nude Mice Bearing Human HepG2 Tumor. CPT-11 (200 mg/kg)
was given intraperitoneally on day 0 only. PHY906 was given orally
30 min before CPT-11 on day 0 and continued twice a day for 8 days
at 500 mg/kg (N=5 in each group).
[0024] FIG. 21. Effect of PHY906 on the Body Weight in CPT-11
Treated NCr-Nude Mice Bearing Human HepG2 Tumor. CPT-11 (200 mg/kg)
was given intraperitoneally on day 0 only. PHY906 was given orally
30 min before CPT-11 on day 0 and continued twice a day for 8 days
at 500 mg/kg (N=5 in each group).
[0025] FIG. 22. Antitumor Effect of L-OddC with PHY906 on Colon 38
Bearing BDF-1 Mice. Female BDF-1 mice (8-10 weeks old, average
weight about 20 g) were injected subcutaneously with Colon 38 tumor
cells. L-OddC (beta-L-Dioxolane-cytidine 25 mg/kg, q.d.X5) was
injected intraperitoneally starting on day zero for 5 consecutive
days. PHY906 was administered orally (1 g/kg, b.i.d.) on day zero
and on a daily basis until the completion of the experiment (q.d.
is an abbreviation for "quaque die" which means once a day, q.d. X5
means each one of five mice received the dose once a day for 5
consecutive days; b.i.d. is an abbreviation for "bis in die", which
means twice a day). (N=5 in each group).
[0026] FIG. 23. Antitumor Effect of VP-16 with PHY906 on Colon 38
Bearing BDF-1 Mice. Female BDF-1 mice (8-10 weeks old, average
weight about 20 g) were injected subcutaneously with Colon 38 tumor
cells. VP-16 (etoposide 25 mg/kg, q.d.X5) was injected
intraperitoneally once a day on day 0 and for 5 consecutive days.
PHY906 was administered orally (1 g/kg, b.i.d.) on day 0 and on a
daily basis until the completion of the experiment. (N=5 in each
group).
[0027] FIG. 24. Antitumor Effect of 5-Fluorouracil (FU) with
PHY906. Five female BDF-1 mice (8-10 weeks old, average weight
about 20 g) were injected subcutaneously with Colon 38 tumor cells.
Only one dose of FU (250 mg/kg) was injected intraperitoneally on
day zero. PHY906 was administered orally (1 g/kg, b.i.d.) on day
zero and on a daily basis until the completion of the experiment.
(N=5 in each group).
[0028] FIG. 25. Antitumor Effect of 5-Fluorouracil (FU) with
PHY906. Female BDF-1 mice in each group (8-10 weeks old, average
weight about 20 g) were injected subcutaneously with Colon 38 tumor
cells. FU (30 mg/kg, q.d. X5) was injected intraperitoneally daily
for 5 consecutive days starting on day 0. PHY906 was administered
orally (1 g/kg, b.i.d.) on day 0 and on a daily basis until the
completion of the experiment. (N=5 in each group).
[0029] FIG. 26. Antitumor Effect of CPT-11 with PHY906 Versus
Loperamide on Colon 38 Bearing BDF-1 Mice. Female BDF-1 mice (8-10
weeks old, average weight about 20 g) were injected subcutaneously
with Colon 38 tumor cells. Mice either received no treatment,
PHY906 alone, CPT-11 alone, CPT-11 and PHY906, or loperamide alone.
The PHY906 and CPT-11 were administered as set forth in FIG. 15.
Only one dose of loperamide was given orally (2 mg/kg, p.o. (oral
administration), b.i.d.) on day zero. (N=5 in each group).
[0030] FIG. 27. Effect of PHY906 (500 mg/kg, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (150 mg/kg, qd, D1 and 8)-Treated
C57/BL6 Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (150 mg/kg)
was given intraperitoneally once a day on days 1 and 8. PHY906 (500
mg/kg) was given orally 30 min before gemcitabine twice a day on
days 1-4 and days 8-11 (N=5 in each group).
[0031] FIG. 28. Effect of PHY906 (500 mg/kg, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (300 mg/kg, qd, D1 and 8)-Treated
C57/BL6 Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (300 mg/kg)
was given intraperitoneally once a day on days 1 and 8. PHY906 (500
mg/kg) was given orally 30 min before gemcitabine twice a day on
days 1-4 and days 8-11 (N=5 in each group).
[0032] FIG. 29. Effect of PHY906 (500 mg/kg, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (600 mg/kg, qd, D1 and 8)-Treated
C57/BL6 Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (600 mg/kg)
was given intraperitoneally once a day on days 1 and 8. PHY906 (500
mg/kg) was given orally 30 min before gemcitabine twice a day on
days 1-4 and days 8-11 (N=5 in each group).
[0033] FIG. 30. Effect of PHY906 (500 mg/kg, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (300 mg/kg, qd, D1 and 8)-Treated BDF-1
Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (300 mg/kg) was given
intraperitoneally once a day on days 1 and 8. PHY906 (500 mg/kg)
was given orally 30 min before gemcitabine twice a day on days 1-4
and days 8-11 (N=5 in each group).
[0034] FIG. 31. Effect of PHY906 (500 mg/kg, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (600 mg/kg, qd, D1 and 8)-Treated BDF-1
Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (600 mg/kg) was given
intraperitoneally once a day on days 1 and 8. PHY906 (500 mg/kg)
was given orally 30 min before gemcitabine twice a day on days 1-4
and days 8-11 (N=5 in each group).
[0035] FIG. 32. Effect of PHY906 (500 mg/kg, bid, D1-4, 8-11, 26-29
and 33-36) on Tumor Growth in Gemcitabine (300 mg/kg, qd, D1, 8, 26
and 33)-Treated Human PANC-1 Tumor Xenografts. Gemcitabine (300
mg/kg) was given intraperitoneally once a day on days 1, 8, 26 and
33. PHY906 (500 mg/kg) was given orally 30 min before gemcitabine
twice a day on days 1-4, 8-11, 26-29 and 33-36 (N=5 in each
group).
[0036] FIG. 33. Effect of PHY906 (500 mg/kg, bid, D1-4, 8-11, 26-29
and 33-36) on Bodyweight Change in Gemcitabine (300 mg/kg, qd, D1,
8, 26 and 33)-Treated Human PANC-1 Tumor Xenografts. Gemcitabine
(300 mg/kg) was given intraperitoneally once a day on days 1, 8, 26
and 33. PHY906 (500 mg/kg) was given orally 30 min before
gemcitabine twice a day on days 1-4, 8-11, 26-29 and 33-36 (N=5 in
each group).
[0037] FIG. 34. Effect of 906MT (1 g/kg*, bid, D1-4 and 8-11) on
Tumor Growth in Gemcitabine (300 mg/kg, qd, D1 and 8)-Treated BDF-1
Mouse Bearing Mouse PAN02 Tumors. Gemcitabine (300 mg/kg) was given
intraperitoneally once a day on days 1 and 8. 906MT (1 g/kg*) was
given orally 30 min before gemcitabine twice a day on days 1-4 and
days 8-11 (N=5 in each group). 906MT is a traditional formulation
obtained in the market place. Thus, the amount used (1 g/kg*) is
approximate.
[0038] FIG. 35. Pharmacokinetics of (A) Gemcitabine (dFdC) and (B)
Its Metabolite (dFdU) in Plasma. One dose of Gemcitabine (600
mg/kg) or/and PHY906 (500 mg/kg) was given orally. PHY906 was given
30 min before gemcitabine. Each mouse was bled twice at different
time points, three mice at each time point.
[0039] FIG. 36. Antitumor Effect of PHY906 on
Gemcitabine+Oxaliplatin Treated NCr-Nude Mice Bearing Human PANC-1
Tumor. Oxaliplatin (10 mg/kg) was given intraperitoneally 30 min
prior to gemcitabine (300 mg/kg) intraperitoneally once a day on
day 1. PHY906 (500 mg/kg) was given orally 30 min before
oxaliplatin twice a day on days 1-4 (N=5 in each group).
[0040] FIG. 37. Effect of PHY906 on the Tumor Growth in CPT-11
Treated NCr-Nude Mice Bearing Human HepG2 Tumor. CPT-11 (200 mg/kg)
was given intraperitoneally on day 0 only. PHY906 was given orally
30 min before CPT-11 on day 0 and continued twice a day for 8 days
at 500 mg/kg (N=5 in each group).
[0041] FIG. 38. Effect of PHY906 on the Tumor Growth in
Capecitabine Treated NCr-Nude Mice Bearing Human HepG2 Tumor.
Capecitabine (360 mg/kg) was given orally twice a day starting on
day 0 for 14 consecutive days. PHY906 was given orally 30 min
before capecitabine twice a day intermittently on days 0-3 and days
7-10 at 500 mg/kg (N=5 in each group).
[0042] FIG. 39. Antitumor Effect of PHY906 on Doxorubicin Treated
NCr-Nude Mice Bearing Human HepG2 Tumor. Doxorubicin (7.5 mg/kg)
was given intraperitoneally once a day on days 1, 4 and 7. PHY906
(500 mg/kg) was given orally 30 min before doxorubicin twice a day
on days 1-4 and 8-11 (N=5 in each group).
[0043] FIG. 40. Antitumor Effect of PHY906 on Thalidomide Treated
NCr-Nude Mice Bearing Human HepG2 Tumor. Thalidomide (1 g/kg) was
given intraperitoneally once a day continuously on days 1-14.
PHY906 (500 mg/kg) was given orally 3 hours before thalidomide
twice a day on days 1-4 (N=5 in each group).
[0044] FIG. 41. Effect of PHY906 on Anti-Tumor Activity of
Oxaliplatin in Colon 38 Bearing BDF-1 Mice. Oxaliplatin (10 mg/kg,
D1) was given intraperitoneally on day 1. PHY906 was given orally
30 min before oxaliplatin twice a day on days 1-4 at 500 mg/kg (N=5
in each group).
[0045] FIG. 42. Effect of PHY906 on Anti-Tumor Activity of
Oxaliplatin/5-FU/LV in Colon 38 Bearing BDF-1 Mice. 5-FU and LV (50
mg/kg each) were given intraperitoneally on days 1, 2, 8 and 9.
Oxaliplatin (5 mg/kg) was given intraperitoneally on days 1 and 8.
PHY906 was given orally 30 min before 5-FU/LV twice a day on days
1-4 and days 8-11 at 500 mg/kg (N=5 in each group).
[0046] FIG. 43. Kaplan-Meier Survival Plot on Colon 38 Bearing
BDF-1 Mice Treated With (A) Vehicle; (B) Oxaliplatin/5-FU/LV; or
(C)PHY906 plus Oxaliplatin/5-FU/LV. Oxaliplatin (10 mg/kg, D1) and
5-FU/LV (50 mg/kg each, D1-2) were given intraperitoneally starting
on day 1. PHY906 was given orally 30 min before oxaliplatin/5-FU/LV
twice a day on days 1-4 at 500 mg/kg (N=5 in each group).
[0047] FIG. 44. Effect of PHY906 on Bodyweight of Colon 38 Bearing
BDF-1 Mice Treated with Oxaliplatin, 5-FU/LV and
Oxaliplatin/5-FU/LV. Oxaliplatin (5 mg/kg, D1) was given
intraperitoneally on day 1. 5-FU and LV (50 mg/kg each) were given
intraperitoneally on days 1, 2, 8 and 9. PHY906 was given orally 30
min before 5-FU/LV or oxaliplatin twice a day on days 1-4 and days
8-11 at 500 mg/kg (N=5 in each group).
SUMMARY OF THE INVENTION
[0048] The inventors of the present invention have unexpectedly
discovered that the herbal composition PHY906 can be used in
various methods for increasing the therapeutic index of one or more
chemotherapeutic compounds and for modulating hematopoietic
activity. The methods disclosed herein can be used to improve the
quality of life for chemotherapy patients by increasing the
efficacy of chemotherapeutic agents and decreasing the toxicity of
the agents when they are administered with PHY906.
[0049] In one aspect, the present invention provides a composition
comprising: i) a pharmaceutically acceptable carrier; ii) an herbal
preparation comprising Scutellaria, Glycyrrhiza, Ziziphus, and
Paeonia; and iii) one or more chemotherapeutic compounds.
[0050] This invention provides the herbal composition PHY906
combined with a pharmaceutically acceptable carrier and optionally
including one or more chemotherapeutic compounds or antiviral
agents. The four plant species which are chosen to make a
particular formulation of PHY906 are each selected from one of four
different groups of herbs: Scutellaria, Licorice, Peony Alba and
Ziziphi fruit. The herbs are chosen so as to obtain one or more of
the desirable attributes of PHY906, wherein such attributes
include, but are not limited to, increasing the therapeutic index
of one or more chemotherapeutic compounds, enhancing the antitumor
activity of one or more chemotherapeutic compounds or enhancing the
antiviral activity of one or more antiviral agents, modulating
hematopoietic activity, modulating hematological and immunological
activity, and improving the quality of life of a mammal undergoing
chemotherapy or antiviral therapy. In some embodiments, the present
invention provides compositions comprising: i) a pharmaceutically
acceptable carrier; ii) an herbal preparation comprising
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia; and iii) a
chemotherapeutic formulation. In some embodiments, the herbal
preparation consists of Scutellaria baicalensis, Glycyrrhiza
uralensis, Ziziphus jujuba, and Paeonia lactiflora. In some
embodiments, the compositions consist essentially of a
pharmaceutically acceptable carrier and material from a plant
species of each of the following genera of herbs: Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia.
[0051] Chemotherapeutic compounds or agents encompassed by this
invention include, but are not limited to, those useful for
treating cancer, parasitic infections, and microbial
infections.
[0052] The compositions and methods of the present invention are
useful for treating any mammal. More specifically, the methods of
the present invention are useful for treating humans.
[0053] This invention further provides compositions which include a
pharmaceutically acceptable carrier; material or chemical from a
plant species of each of the following genera of herbs:
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia; and one or more
chemotherapeutic compounds. Preferably, the composition comprises a
pharmaceutically acceptable carrier, an herbal preparation
comprising Scutellaria, Glycyrrhiza, Ziziphus and Paeonia, and a
chemotherapeutic formulation comprising one or more
chemotherapeutics or antiviral agents. More preferably, the herbal
preparation comprises material or chemical from Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia. Most preferably, this invention
provides such compositions which include Scutellaria baicalensis,
Glycyrrhiza uralensis, Ziziphus jujuba and Paeonia lactiflora.
[0054] The present invention provides methods of modulating
hematopoietic activity for the treatment of a disease by
administering to a mammal in need of such treatment a
therapeutically effective amount of a composition consisting
essentially of a pharmaceutically acceptable carrier and material
or chemical from or herbal preparation comprising a plant species
of each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia.
[0055] In another aspect, the present invention provides a method
of treating a disease in a mammal in need thereof comprising
administering a therapeutically effective amount of a composition
comprising: i) a pharmaceutically acceptable carrier; ii) an herbal
preparation comprising Scutellaria, Glycyrrhiza, Ziziphus, and
Paeonia; and iii) one or more chemotherapeutic compounds.
[0056] The present invention also provides methods of treating
diseases in mammals in need of such treatment which includes
administering a therapeutically effective amount of a composition
comprising a pharmaceutically acceptable carrier; material or
chemical from, or herbal preparation comprising a plant species of
each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia; and one or more chemotherapeutic
compounds.
[0057] The present invention further provides methods of treating
diseases in a mammal in need of such treatment which includes
administering a therapeutically effective amount of one or more
chemotherapeutic compounds or antiviral agents and a composition
which includes a pharmaceutically acceptable carrier; material or
chemical from, or herbal preparation comprising a plant species of
each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia. The present invention includes such methods
wherein the composition is administered before the administration
of the one or more chemotherapeutic compounds. The present
invention also includes such methods wherein the composition is
administered after the administration of the one or more
chemotherapeutic compounds
[0058] The present invention provides methods for increasing the
therapeutic index of cancer therapeutic compounds used in the
treatment of cancer. The present invention also provides methods
for increasing the therapeutic index of antiviral agents used in
the treatment of antiviral diseases. More specifically, the present
invention provides such methods which include administering one or
more anticancer or antiviral agent in combination with a
therapeutically effective amount of a composition comprising a
pharmaceutically acceptable carrier and material or chemical from,
or herbal preparation comprising, a plant species of each of the
following genera of herbs: Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia. The methods of the present invention provide the use of
material or chemical from, or herbal preparation comprising, such
herbs which is in the form of a granulated extract from an aqueous
solution that includes but is not limited to water and alcohol.
Such compositions can be in an ingestible form, such as, but not
limited to, powders, capsules, liquids and tablets. Alternatively,
the methods of the present invention use such compositions in the
form of a suppository.
[0059] In another aspect, the present invention provides a method
of increasing the therapeutic index of cancer therapeutic compounds
for the treatment of cancer by administering to a mammal in need
thereof a therapeutically effective amount of a composition
comprising a pharmaceutically acceptable carrier, and an herbal
preparation comprising Scutellaria, Glycyrrhiza, Ziziphus, and
Paeonia.
[0060] In yet another aspect, the present invention provides a
method of relieving side effects of a chemotherapeutic compound in
a mammal comprising administering a composition comprising: i) a
pharmaceutically acceptable carrier; ii) an herbal preparation
comprising Scutellaria, Glycyrrhiza, Ziziphus, and Paeonia; and
iii) one or more chemotherapeutic compounds.
[0061] In yet another aspect, the present invention provides a
method of improving the quality of life of a mammal undergoing
chemotherapy which comprises administering a therapeutically
effective amount of one or more chemotherapeutic compounds and a
composition comprising: i) a pharmaceutically acceptable carrier;
ii) an herbal preparation comprising Scutellaria, Glycyrrhiza,
Ziziphus, and Paeonia; and iii) one or more chemotherapeutic
compounds.
[0062] The present invention also provides methods of improving the
quality of life of a mammal undergoing chemotherapy or antiviral
therapy which comprises administering a therapeutically effective
amount of one or more chemotherapeutic compounds and a composition
comprising:
i) a pharmaceutically acceptable carrier; and ii) material or
chemical from a plant species of each of the following genera of
herbs: Scutellaria, Glycyrrhiza, Ziziphus and Paeonia.
[0063] The present invention contemplates administering a
chemotherapeutic formulation comprising one or more
chemotherapeutic agents in combination with a composition
comprising a pharmaceutically acceptable carrier and an herbal
preparation comprising a plant species of each of the following
genera of herbs: Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia.
[0064] The present invention uses the disclosed herbal compositions
for increasing the antitumor activity of chemotherapeutic agents,
increasing the antiviral activity of antiviral agents, decreasing
the toxicity of the chemotherapeutic or antiviral agent, modulating
the hematological and immunological activity of a mammal, and
improving the quality of life of a mammal undergoing chemotherapy
or antiviral therapy.
[0065] In one aspect, the present invention discloses a method of
treatment comprising a chemotherapeutic regimen comprising one or
more chemotherapeutic compounds and a composition comprising a
pharmaceutically acceptable carrier and an herbal preparation
comprising Scutellaria, Glycyrrhiza, Ziziphus and Paeonia. In
another aspect, the present invention discloses a method of
treatment comprising an antiviral regimen comprising one or more
antiviral agents and a composition comprising a pharmaceutically
acceptable carrier and an herbal preparation comprising
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia.
[0066] Further, the present invention provides a therapeutic
regimen comprising one or more chemotherapeutic or antiviral
compound and a composition comprising a pharmaceutically acceptable
carrier and an herbal preparation comprising Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia.
[0067] Additionally, the present invention discloses
chemotherapeutic regimens and compositions comprising three
chemotherapeutic compounds, preferably, CPT-11/FU/LV or
oxaliplatin/CPT-11/FU/LV and an herbal preparation comprising
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia or material or
chemical from a plant species of each of the following genera of
herbs: Scutellaria, Glycyrrhiza, Ziziphus and Paeonia. The present
invention contemplates antiviral therapies comprising one or more
antiviral agents.
DETAILED DESCRIPTION OF THE INVENTION
[0068] The present invention is based in part on the finding that
PHY906 reduced chemotherapy-induced toxicities including body
weight loss and mortality, and that PHY906 also enhanced the
antitumor efficacy of a broad-spectrum of anticancer agents, such
as CPT-11, 5-FU, VP-16, L-OddC, capecitabine, gemcitabine,
oxaliplatin, doxorubicin, thalidomide, CPT-11/5-FU/LV,
gemcitabine/oxaliplatin, and oxaliplatin/5-FU/LV in in vivo animal
models. PHY906 is a traditional Chinese botanical formulation
consisting of 4 different herbs, and it has been used for some 1800
years to treat gastrointestinal ailments, some of which are
commonly observed side effects in cancer patients undergoing
chemotherapy.
[0069] The present invention is also based in part on the finding
that PHY906 significantly enhanced the therapeutic index of
chemotherapeutic agents studied in both hepatocellular and
pancreatic cancer models. PHY906 was co-administered with either
the oral 5-FU prodrug capecitabine, CPT-11, doxorubicin, or
thalidomide in human hepatocellular xenografts mouse model, and
with gemcitabine, oxliplatin, gemcitabine/oxaliplatin in mouse
pancreatic cancer model. Co-administration of PHY906 with either
capecitabine or gemcitabine in animal models did not alter the
pharmacokinetic profile of capecitabine, gemcitabine, or their
respective metabolites.
[0070] The effect of PHY906 was also examined on the antitumor
activities of oxaliplatin, 5-FU/LV, and oxaliplatin/5-FU/LV in
colorectal cancer mouse models. The present invention is based in
part on the finding that PHY906 enhanced the antitumor activities
of oxaliplatin, and oxaliplatin/5-FU/LV as shown in FIGS. 41 and
42. PHY906 not only reduced the mortality to 60%, but also delayed
the onset of animal death caused by oxaliplatin/5-FU/LV, as shown
in FIG. 43.
[0071] These preclinical in vivo studies have provided the
rationale for developing PHY906 in the clinical setting. A phase
I/IIA double-blind, placebo-controlled, dose-escalation study of
PHY906 was initiated to evaluate the potential effect of PHY906 in
modulating CPT-11-induced diarrhea associated with the bolus,
weekly schedule of CPT-11/5-FU/LV in advanced colorectal cancer
patients. A second phase I/II open-label, dose-escalation clinical
trial has been opened to patient accrual to evaluate the role of
PHY906 in combination with capecitabine in the treatment of
hepatocellular carcinoma.
[0072] PHY906 has also been shown in mouse models to enhance the
therapeutic index of several anticancer agents such as irinotecan
(CPT-11), 5-fluorouracil (5-FU),
irinotecan/5-fluorouracil/leucovorin (LV), etoposide (VP-16) and
L-OddC by both potentiating the antitumor effects of the
therapeutic agents as well as reducing various host toxicities. To
investigate whether there is a need of all four botanicals to
achieve the full observed biological activity, five formulations
were studied: the complete formulation with all four botanicals and
each of four formulations missing one of the botanical ingredients
(A=Scutellaria, B=P aeonia, C=Ziziphus and D=Glycyrrhiza). In the
BDF-1 mice bearing Colon 38 tumor model, the potentiation of the
antitumor effect of CPT-11 was impaired when either botanical A or
botanical B was removed from the PHY906 formulation. In contrast,
botanicals C and D were not found to contribute to the potentiation
of the antitumor effect of CPT-11. However, when examining weight
loss as an undesired toxic side effect of CPT-11, all of the
botanicals except for B, contributed in protecting against body
weight loss. Thus, the biological activity of PHY906 observed
requires all four botanicals.
[0073] In summary, the present invention is based in part on the
following findings:
1. In different mouse models, PHY906 increased the therapeutic
index of various types of chemotherapeutic agents in human
pancreatic cancer, hepatocellular carcinoma, and colorectal cancer.
Specifically, PHY906 increased the therapeutic index of a broad
spectrum of chemotherapeutic agents by enhancing the antitumor
activity, decreasing the mortality, and protecting against the body
weight loss. 2. PHY906 did not alter the pharmacokinetics of these
chemotherapeutic agents in mouse models. 3. The mechanisms of drug
action of PHY906 in the potentiation of studied cancer
chemotherapeutic agents can be attributed but not limited to the
inhibition on NF-.kappa.B, MMPs, or to the destruction of the
integrity of hepatoma sinusoids. 4. All four herbs (A=Scutellaria,
B=P aeonia, C=Ziziphus and D=Glycyrrhiza) were required in PHY906
formula for the full range of observed efficacy: Herb A contributed
to both body weight loss protection and antitumor potentiatiation
(p<0.05) in the PHY906 formula. Herb B contributed to the
enhancement of antitumor activity of PHY906 (p<0.05), but not to
the protection of cancer chemotherapy drug-induced body weight
loss. Herbs C and D protected against the body weight loss induced
by cancer chemothreapy agents. 5. Different batches of PHY906
showed consistency in both human CYP450 inhibition and cell growth
inhibition: Herb A was a potent inhibitor for human CYP450 as well
as for cell growth. Herb D showed very low cytotoxicity in the cell
lines examined and is a very weak inhibitor for human CYP450. 6.
PHY906 showed strong inhibitory activity against human CYP450 in
vitro. However, animal data revealed that PHY906 did not affect the
CPT-11 metabolism or impair the anticancer activity of
chemotherapeutic agent.
[0074] Cancer chemotherapeutic agents often induce severe adverse
side effects that can affect patients' quality of life, as well as
interfere with the therapeutic regimen. The present invention is
based in part on the discovery that Chinese herbal medicines in
combination with standard anticancer agents are useful for reducing
the adverse side effects of cancer chemotherapeutic agents and for
improving the quality of life of patients undergoing chemotherapy.
PHY906, a botanical formulation composed of four distinct herbs,
has been used for centuries for the treatment of various
gastrointestinal ailments and other illnesses in China. The present
invention is based on the results of experiments performed in
animal models evaluating the potential efficacy of PHY906 in
relieving side effects induced by cancer chemotherapy agents in
colorectal cancer patients. Specifically, the present invention is
based in part on the finding that PHY906 reduces various host
toxicities induced by CPT-11, FU, FU/LV, CPT-11/FU/LV, L-OddC,
VP-16, thalidomide, doxorubicin, capecitabine, gemcitabine,
oxaliplatin, gemcitabine/oxliplatin or oxaliplatin/FU/LV treatment,
as well as maintaining and even potentiating the antitumor activity
of chemotherapeutic agents. More specifically, PHY906 enhances the
therapeutic index of CPT-11, 5-FU, capecitabine, oxaliplatin,
doxorubicin, thalidomide, VP-16, L-OddC, and the triple
combinations of CPT-11/FU/LV and oxaliplatin/FU/LV by both
potentiating antitumor effects of the therapeutic agents and
reducing various host toxicities. These findings are not limited to
one specific anticancer agent or one specific tumor model.
[0075] The present invention is also based in part on the discovery
of a regimen that can be used in conjunction with various
anticancer agents to lower the dose limiting toxicity and increase
efficacy of the agents. This discovery is an extremely important
addition to the armamentarium of treatment approaches for human
cancers.
DEFINITIONS
[0076] Unless defined otherwise, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. Although
any methods and materials similar or equivalent to those described
herein can be used in the practice or testing of the present
invention, the preferred methods and materials are described.
[0077] As used herein, "cancer chemotherapeutic" or "cancer
chemotherapeutic agent" refers to chemotherapeutic agent for the
treatment of neoplastic disease or cancer.
[0078] As used herein, "chemotherapy" refers to treatment of
disease by means of chemical substances or drugs.
[0079] As used herein, the term "chemotherapeutic formulation"
refers to a composition comprising a chemotherapeutic agent.
[0080] As used herein, the term "extract" refers to a concentrated
preparation of a vegetable or animal drug obtained by removing the
active constituents therefrom with a suitable menstruum (solvent),
evaporating all or nearly all the solvent and adjusting the
residual mass or powder to a prescribed standard. Extracts are
prepared in three forms, semiliquid or of syrupy consistency,
pilular or solid, and as dry powder (see
http://www.graylab.ac.uk/cgi-bin/omd?query=extract).
[0081] In one embodiment, extracts are concentrated forms of crude
drugs used in a variety of solid and semisolid dosage forms (in
Remington's Pharmaceutical Sciences 17th ed. (Gennaro, ed), Chapter
84, pp. 1516-1517, Mack Publishing Co, Easton, Pa. (1985)). For
example, pilular (i.e., plastic masses) extracts are of a
consistency where they are suitable for pill masses and are made
into pills (e.g., pure Glycyrrhiza extract USP). Further, pilular
masses are well suited for use in ointments and suppositories.
Powdered extracts are better suited for powdered formulations such
as capsules, powders and tablets. Further, semiliquid or extracts
of syrupy consistency can be used in the manufacture of
pharmaceutical preparations (Remington's Pharmaceutical Sciences,
1985).
[0082] In a related aspect, extracts can be considered solutions of
active ingredients obtained by soaking or steeping preparations of
vegetable or animal crude drugs in liquids (maceration) or by
passing such crude drugs through porous substances (percolation)
for use as a medicinal agent. Further, medicinal agents of this
type may be in the form of tinctures or fluidextracts [sic]
(Remington's Pharmaceutical Sciences, 1985).
[0083] In one embodiment, the extract is in tincture form. For
example, tinctures may include, but are not limited to, alcoholic
or hydroalcoholic solutions prepared from vegetable matter or from
chemical substances. Tinctures may be made by either percolation or
maceration and are traditionally assigned potency by the amount of
activity of a specified weight of the drug (in grams) per 100 ml of
tincture (Remington's Pharmaceutical Sciences, 1985). For example,
Sweet Orange Peel Tincture contains 50 g of sweet orange peel per
100 ml of tincture.
[0084] In another embodiment, the extract is in fluid extract [sic]
form. For example, fluid extracts include, but are not limited to,
liquid preparations of vegetable drugs comprising alcohol as the
solvent or as a preservative, or both, where traditionally each ml
contains the therapeutic constituents of 1 gram of the drug that it
represents. Fluid extracts can be made by percolation as a general
method (Remington's Pharmaceutical Sciences, 1985).
[0085] As used herein, the term "hematological activity" refers to
activity associated with blood and blood forming organs.
[0086] Technically speaking, the term "herb" refers to a small,
non-woody (i.e., fleshy stemmed), annual or perennial seed-bearing
plant in which all the aerial parts die back at the end of each
growing season. Herbs are valued for their medicinal, savory or
aromatic qualities. As the word is more generally used and as the
word is used herein, an "herb" refers to any plant or plant part
which has a food supplement, medicinal, drug, therapeutic, or
life-enhancing use. Thus, as used herein, an herb is not limited to
the botanical definition of an herb but rather to any botanical,
plant or plant part used for such purposes, including any plant or
plant part of any plant species or subspecies of the Metaphyta
kingdom, including herbs, shrubs, subshrubs, and trees. Plant parts
used in herbal compositions include, but are not limited to, seeds,
leaves, stems, twigs, branches, buds, flowers, bulbs, corns,
tubers, rhizomes, runners, roots, fruits, cones, berries, cambium
and bark.
[0087] As used herein, an "herbal composition or formulation"
refers to any composition or formulation which includes herbs,
herbal plants, herbal plant parts and/or herbal extracts. Thus, as
used herein, an herbal composition or formulation includes herbal
preparation comprising herbal food supplements, herbal medicines,
herbal drugs, and medical foods. Examples of herbal compositions
include, but are not limited to, the following components: a whole
plant or a plant part of a single plant species; whole plants or
plant parts of multiple plant species; multiple components derived
from a single plant species; multiple components derived from
multiple plant species; herbal extracts; or any combination of
these various components. Also contemplated are herbal compositions
comprising one or more chemicals derived from a single or multiple
plant species.
[0088] For a thorough review of various herbal compositions, see,
for example, Kee Chang Huang, The Pharmacology of Chinese Herbs,
CRC Press (1993), herein incorporated in its entirety.
[0089] As used herein, the term "immunological activity" refers to
activity associated with the immune system, immunity, induced
sensitivity, and allergy.
[0090] As used herein, the term "mortality rate" refers to the
proportion of deaths in a population or to a specific number of the
population, where mortality is defined as the death rate or ratio
of the total number of deaths to the total population. For example,
the 30 day mortality rate after ischemic stroke symptom onset can
vary from about 13.3% (e.g., after treatment with tissue type
plasminogen activator, see Albers et al., JAMA (2000)
283(9):1145-1150) to greater than about 65% (e.g., hemorrhage
stroke, see Mahaffey et al., Am Heart J (1999) 138(3 Pt
1):493-499).
[0091] As used herein, the term "Quality of life (QOL)" refers to
the general well-being of an animal, especially a mammal, even more
specifically a human. The QOL of an individual can be evaluated
based on any one parameter, a group of two or more parameters or on
a general overall evaluation or score. Examples of useful indices
for evaluating QOL include, but are not limited to, those
associated with sleeping patterns; eating patterns; drinking
patterns; agility; mobility; skin tone; vision; hair
retention/loss/growth; muscle tone; muscle mass; strength; weight;
sinus health; presence, absence or degree of inflammation; feelings
of discomfort; ability to accomplish specific tasks; anxiety
levels; response times; ability to concentrate; memory retention;
verbal ability; sound perception; presence, absence or degree of
headaches; muscle spasms; nerve damage; taste; touch; smell;
presence or absence of opportunistic diseases; and presence or
absence of parasites.
[0092] As used herein, the term "regimen" refers to a program of
treatment.
[0093] As used herein, the term "therapeutic index" refers to how
selective a drug is in producing the desired effects. Therapeutic
index is the ratio of LD.sub.50 to ED.sub.50. ED.sub.50, the median
effective dose, is the dose of a drug required to produce a
specified effect in 50% of the population. LD.sub.50 is the median
lethal dose as determined in experimental animals.
Specific Embodiments
[0094] Chemotherapy
[0095] In general, chemotherapy refers to the treatment of disease,
especially neoplasms, parasitic infections and microbial diseases,
with chemical agents that in some manner act on the infective
organisms or tumors.
[0096] Cancer Chemotherapy
[0097] Introduction: Chemotherapy continues to be one of the most
effective modalities for treating cancer in patients. Although
quite effective, chemotherapeutic agents are also well known to
adversely disrupt the quality of life of patients. Some commonly
observed side effects include myelosuppression and
immunosuppression, diarrhea, peripheral neuropathy, nausea and
vomiting, fever, liver dysfunction and cardiac toxicity, etc.
(Physicians Desk Reference (1999) Medical Economics Company). In
many instances, these adverse side effects prevent patients from
receiving escalating doses or additional courses of therapy,
thereby comprising the efficacy of these agents. Alleviation of
some or all of these side effects, without compromising the
anticancer activity of chemotherapeutic agents, would not only
improve the quality of life (QOL) of cancer patients, but also
allow for a more aggressive treatment protocol, resulting in
possibly improved clinical success. Currently, most supportive
therapies use single agents, such as anti-emetics, anti-mucositis
agents, and colony growth factors, that target individual side
effects, but do not address the broad spectrum of side effects
associated with cancer chemotherapy (Bleiberg H and Cvitkovic E.,
Eur J Cancer 32A(Suppl 3):518-S23 (1996); Wierda D. and Matamoros
M., Toxicol & Applied Pharmacol 75:25-34 (1984); Goldber R. M.
and Erlichman C., Oncology 12: 59-63 (1988)).
[0098] Drugs for treating cancer include the more conventional
natural products such as paclitaxel (TAXOL.RTM.), the
semisynthetics such as etoposide, and many newer, diverse agents
such as interleukin-2 and all-trans-retinoic acid. For a
comprehensive list of chemotherapeutic agents useful in treating
neoplastic diseases, see, for example, Table X-1 at pages 1227-1229
of Calabresi and Chabner (1996).
[0099] The major adverse effects associated with commonly
administered cancer chemotherapies are provided in Table 1.
TABLE-US-00001 TABLE 1 Major Adverse Effects of Cancer Chemotherapy
Major Adverse Health Effects Antineoplastic Agent Pancreatitis
VP-16, ara C Alopecia VP-16, Doxorubicin, Taxol, FU, araC
Cardiotoxicity Taxol, Doxorubicin Cutaneous Doxorubicin Diarrhea
CPT-11 Dyspnea ara C Flush Tamoxifen Fever/Chills VP-16,
Doxorubicin Hepatotoxicity VP, Taxol, ara C, Methotrexate
Nephrotoxicity Cisplatin Ototoxicity Cisplatin Bone Marrow
Hypoplasia Almost all anticancer drugs
[0100] The fluoropyrimidine analog, 5-fluorouracil (5-FU or FU),
exhibits a broad spectrum of clinical activity. It remains one of
the most active agents in the treatment of colorectal cancer both
in the adjuvant and advanced disease setting, and in other G1
malignancies as well (Pinedo and Peters, 1988). In addition, this
agent is active against cancers of the breast, and head and
neck.
[0101] Recent advances in the therapy of colorectal cancer have
used biochemical modulation to selectively activate specific
pyrimidine metabolic pathways. The reduced folate, leucovorin (LV),
is an effective biochemical modulator and has been used in clinical
treatments in combination with FU (Peters and Van Groeningen, 1991;
Joulia, et al., 1999). It has been shown that the addition of
exogenous folate in the form of LV enhances responses to FU in
clinical trials (Calabresi and Chabner, Page 1250, 1996). The
purported mechanism of interaction of LV is enhanced thymidylate
synthase inhibition.
[0102] The response rate to FU in patients with advanced disease is
improved from 10%-12% (FU treatment alone) to 20%-30% (FU/LV
treatment).
[0103] Gemcitabine is the only clinically approved chemotherapeutic
agent for pancreatic cancer; however, the response rate in patients
to gemcitabine is only 6-11% and the overall survival time is
generally 4-6 months. Gemcitabine is a nucleoside analog with two
mechanisms of action, including the inhibition of ribonucleotide
reductase, an enzyme that converts nucleotide diphosphate to
deoxynucleotide triphosphate and that is required for DNA synthesis
and that competes with deoxycytidine triphosphate as a fraudulent
base in DNA synthesis (3, 5-10). With the low response and survival
rates of gemcitabine monotherapy, several gemcitabine-combination
drug regimens have been tested clinically for improving therapeutic
efficacy. These trials include gemcitabine with other commonly used
and FDA-approved anti-cancer drugs including CPT-11, capecitabine,
and oxaliplatin (11-14). Unfortunately, no satisfactory combination
drug regimens have been discovered and an effective regimen for
pancreatic cancer is urgently needed.
[0104] Gemcitabine: Gemcitabine (GEMZAR.RTM.) was approved by the
FDA in 1996 for the treatment of locally advanced or metastatic
pancreatic cancer, either as initial therapy or following treatment
with the chemotherapy agent fluorouracil. It also has been approved
for the treatment of locally advanced or metastatic non-small cell
lung cancer, in combination with the chemotherapy agent cisplatin
(PLATINOL.RTM.). Gemcitabine is administered intravenously (i.v.)
and the dose depends on several factors, including the condition
being treated, the size of the patient, the particular treatment
regimen being used, and the overall health of the patient.
[0105] Gemcitabine is the generic name assigned to
2'-deoxy-2',2'-difluoro-cytidine. It is also known chemically as
1-(4-amino-2-oxo-1H-pyrimidin-1-yl)-2-desoxy-2,2-difluororibose.
Gemcitabine is disclosed in U.S. Pat. Nos. 4,808,614 and 5,464,826,
which are incorporated herein by reference for their teaching of
how to synthesize, formulate, and use gemcitabine for treating
susceptible neoplasms. Gemcitabine is commercially available as the
monohydrochloride salt, and as the .beta.-isomer.
[0106] Gemcitabine belongs to a group of drugs called
antimetabolites. Generally, antimetabolites produce their
anti-cancer effects by inhibiting the ability of a cell to produce
or repair DNA, thereby making the cell unable to replicate or
repair itself and ultimately causing cellular death. Gemcitabine
exhibits cell phase specificity, primarily killing cells undergoing
DNA synthesis (S-phase) and also blocking the progression of cells
through the G1/S-phase boundary. Gemcitabine is metabolized
intracellularly by nucleoside kinases to the active diphosphate
(dFdCDP) and triphosphate (dFdCTP) nucleosides. The cytotoxic
effect of gemcitabine is attributed to a combination of two actions
of the diphosphate and the triphosphate nucleosides, which leads to
inhibition of DNA synthesis. First, gemcitabine diphosphate
inhibits ribonucleotide reductase, which is responsible for
catalyzing the reactions that generate the deoxynucleoside
triphosphates for DNA synthesis. Inhibition of this enzyme by the
diphosphate nucleoside causes a reduction in the concentrations of
deoxynucleotides, including dCTP. Second, gemcitabine triphosphate
competes with dCTP for incorporation into the DNA. The reduction in
the intracellular concentration of dCTP (by the action of the
diphosphate) enhances the incorporation of gemcitabine triphosphate
into the DNA (self-potentiation). After the gemcitabine nucleotide
is incorporated into the DNA, only one additional nucleotide is
incorporated into the DNA. After this addition, there is inhibition
of further DNA synthesis.
[0107] Like other chemotherapeutic agents, gemcitabine causes
unwanted side effects. The more common side effects of using
gemcitabine include but not limited to decreased white blood cell
count with increased risk of infection, decreased platelet count
with increased risk of bleeding, nausea, vomiting, increased liver
function blood tests, and fatigue. The less common side effects are
diarrhea; sores in mouth or on lips; flu-like symptoms; skin rash;
swelling of hands, ankles, and face; hair thinning; and
itching.
[0108] Oxaliplatin: Oxaliplatin (ELOXATIN.RTM.) was approved in
August 2002 by the FDA for second-line treatment of patients with
metastatic carcinoma of the colon and rectum. In February 2005, it
was approved for first-line treatment of advanced colorectal
cancer. This recent approval recommends use of oxaliplatin in
combination with 5-fluorouracil and leucovorin (5-FU/LV).
Oxaliplatin is administered intravenously, and the dose depends on
several factors, including the condition being treated, the size of
the patient, the particular regimen being used and the overall
health of the patient.
[0109] Oxaliplatin is a diaminocyclohexane compound that is known
to cause DNA damage of the same sites of adduct formation as does
cisplatin. It is also known as L-OHP and is a third generation
platinum complex. The chemical name of oxaliplatin as
cis-oxalato(trans-1-1,2-cyclohexane-diamine)platinum (II).
Oxaliplatin is described in U.S. Pat. No. 4,169,846 and in related
U.S. Pat. No. 5,290,961; U.S. Pat. No. 5,298,642; U.S. Pat. No.
5,338,874; U.S. Pat. No. 5,420,319 and International Patent
Application No. PCT/IB/00614, which are herein incorporated by
reference in their entirety.
[0110] The term "oxaliplatin" as used herein includes
cis-oxalato(trans-1-1,2-diaminocyclohexane)platinum(II), its optic
enantiomer cis-oxalato(trans-d-1,2-diaminocyclohexane)platinum(II)
and any racemic mixture thereof. The term "oxaliplatin" also
includes cis-oxalato (trans-1-1,2-diaminocyclohexane) platinum (II)
having high optical purity, namely an optical purity equal to or
higher than about 99.5%, for example a cis-oxalato
(trans-1-1,2-diaminocyclohexane) platinum(II), wherein the melting
point is between about 198.degree. C. and about 292.degree. C.,
obtained following the procedure described in U.S. Pat. No.
5,338,874 (which is incorporated by reference in its entirety) and,
especially, a
cis-oxalato(trans-1-1,2-cyclohexanediamine)platinum(II), which
possesses optical purity of about 99.94% or more and a melting
point between about 198.3.degree. C. and about 199.7.degree. C.,
obtained following the procedure disclosed in U.S. Pat. No.
5,420,319, which is incorporated by reference in its entirety.
Derivatives of oxaliplatin include, but are not limited to,
carboplatin and cisplatin.
[0111] Oxaliplatin belongs to a new class of platinum agent. It
contains a platinum atom complexed with oxalate and
diaminocyclohexane (DACH). The bulky DACH is thought to contribute
greater cytotoxicity than cisplatin and carboplatin (Wiseman et
al., Drugs Aging 1999; 14(6):459-75). The exact mechanism of action
of oxaliplatin is not known. Oxaliplatin forms reactive platinum
complexes which are believed to inhibit DNA synthesis by forming
interstrand and intrastrand cross-linking of DNA molecules.
Oxaliplatin is not generally cross-resistant to cisplatin or
carboplatin, possibly due to the DACH group and resistance to DNA
mismatch repair (Wiseman et al., Drugs Aging 1999, 14(6):459-75;
Misset et al., Critical Reviews in Oncology-Hematology 2000,
35(2):75-93). Preclinical studies have shown oxaliplatin to be
synergistic with fluorouracil and SN-38, the active metabolite of
irinotecan (Cvitkovic et al., Semin Oncol 1999, 26(6):647-62).
Oxaliplatin is a radiation-sensitizing agent (Freyer et al., Proc
Am Soc Clin Oncol 2000, 19:260a; Carraro et al., Proc Am Soc Clin
Oncol 2000, 19:291 a). It is cell-cycle-phase nonspecific
(Sanofi-Synthelabo. ELOXATIN.RTM.: Summary of product
characteristics (Europe): France; 1 Oct. 1999).
[0112] Like other chemotherapeutic agents, there are side effects
with the use of oxaliplatin. The more common side effects include
nausea, vomiting, numbness and tingling in hands and/or feet due to
nerve irritation, numbness of lips, diarrhea, abdominal pain, mouth
sores, difficulty breathing, and fatigue. The less common side
effects include difficulty walking, decreased white blood cell
count with increased risk of infection, decreased platelet count
with increased risk of bleeding, difficulty swallowing or
breathing, poor tolerance to cold temperatures, and allergic
reaction with rash, itching, swelling lips or tongue, or sudden
cough.
[0113] Thalidomide: Thalidomide, a derivative of glutamic acid, was
first prescribed as an antiemetic drug in 1950s. Unfortunately, it
was found that thalidomide had teratogenic effects in developing
fetuses. Consequently, thalidomide was hauled from the market. In
recent years, thalidomide has been investigated in other areas of
medicine. It was found that thalidomide significantly decreased the
formation of new vessels and inhibited vascular endothelial growth
factor (VEGF)-induced angiogenesis, an important progression of
haematological malignancies. Several clinical trials implied that
thalidomide might also possess antineoplastic properties.
[0114] Thalidomide exists in two isomeric forms which are
designated as right-handed and left-handed. These two isomers
spontaneously interchange forming a racemic mixture. Thalidomide
derived analogs include the Immunomodulatory Drugs (IMiDs) and
SelCIDs (Selective Cytokine Inhibitory Drugs). Examples of IMiDs
include but are not limited to CC-4047 and CC-5013. Examples of
SelCIDs include but are not limited to CC-7034 and CC-9088. Other
derivatives of thalidomide include but are not limited to
lenalidomide, .alpha.-EM12, .beta.-EM12, and
phthalimidophthalimide.
[0115] In recent years, thalidomide has been found to have some
effect in the treatment of various diseases such as but not limited
to AIDS, leprosy, rheumatoid arthritis, and cancer. Specifically,
in HIV patients, thalidomide has been helpful in reducing the
effects of cachexia (progressive loss of body and muscle mass) and
in treating aphothous ulcers (growth of sores in mouth and throat).
Thalidomide is also effective in stopping HIV replication.
Thalidomide has also been shown to have anti-inflammatory and
immunosuppressant qualities which makes it potentially effective in
the treatment of Behcet's disease, sarcoidosis, scleroderma, and
Crohn's disease.
[0116] Moreover, thalidomide has been established as an effective
therapy for the treatment of radvanced relapsed and refractory
multiple myeloma (MM) (Singhal et al. 1999 N. Engl. J. Med.
341:1565-1571). Studies have demonstrated that both thalidomide and
IMiDs not only act to inhibit angiogenesis, but also act directly
to induce both apoptosis and growth arrest in resistant myeloma
cells. They also block both the adhesion of myeloma cells to bone
marrow stromal cells and the related protection against apoptosis,
and block the increased secretion of myeloma cell growth, survival,
and migratory factors such as interleukin-6, tumor necrosis factor
alpha (TNF-.alpha.), and vascular endothelial growth factor,
triggered by the binding of myeloma cells to bone marrow stromal
cells. In addition, they expand natural killer cell and T-cell
numbers, and improve function against human myeloma cells and
enhance their susceptibility to antibody-dependent cell-mediated
cytotoxicity in vivo. (See for example, Anderson et al. 2001 Semin
Hematol 38:6-10; Hideshima et al. 2000 Blood 96:2943-2950; Gupta et
al 2001 Leukemia 15:1950-1961; Davies et al. 2001 Blood 98:210-216;
Treon et al. 2001 J Immunother 24:236-271; Lentzsch et al. 2003
Leukemia 17:41-44). However, the efficacy of thalidomide has been
significantly limited by adverse effects, which include sedation,
neuropathy, constipation, and deep vein thrombosis).
[0117] Doxorubicin: Doxorubicin (Adriamycin, DOX, ADM) is an
anthracycline antibiotic produced by the fungus Streptomyces
peucetius and has been approved for the therapy of various cancers.
Doxorubicin is an inhibitor of DNA topoisomerase II; it damages DNA
by the intercalation of the anthracycline portion, metal ion
chelation, or by the generation of free radicals. Although there is
no FDA-approved chemotherapeutic agent available for the treatment
of hepatocellular carcinoma, doxorubicin has been used a current
"standard" chemotherapy. A response rate of 16% (range 0-35%) was
reported in eight trials that included 475 evaluable patients
(Engstrom P F et al., (1993) in Cancer Medicine Holland J F, Frei E
(eds) pp. 1430-1441. London:Lea and Febiger).
[0118] Capecitabine (XELODA.RTM.), an oral fluoropyrimidine, is a
rationally designed oral prodrug efficiently absorbed from the
gastrointestinal tract and converted to 5-FU, preferentially in
neoplastic tissues. It has been approved by the FDA as a first-line
chemotherapy for the treatment of colorectal and breast cancers
with reduced toxicities (15-17). Capecitabine has also shown
promising antitumor activity as a single agent in pancreatic cancer
(18) and liver cancer (19).
[0119] Capecitabine is an oral medication used for the treatment of
cancer. Capecitabine was approved by the FDA in 1998 for the
treatment of metastatic breast cancers that are resistant to other
medicines such as paclitaxel (TAXOL.RTM.), doxorubicin, and
adriamycin. In 2001, it was approved by the FDA for the treatment
of metastatic colorectal cancer, which is frequently treated with
5-FU. It generally is administered for 14 days, followed by a 7-day
rest period during each 21-day cycle.
[0120] Capecitabine is a fluoropyrimidine carbamate with
antineoplastic activity. The chemical name for capecitabine is
5'-deoxy-5-fluoro-N-[(pentyloxy)carbonyl]-cytidine. Capecitabine is
described in U.S. Pat. No. 5,472,949, which is herein incorporated
by reference in its entirety.
[0121] Capecitabine belongs to a group of chemotherapeutic agents
known as antimetabolites. It is a prodrug of
5'-deoxy-5-fluorouridine (5'-DFUR) with an unique mechanism of
activation. Capecitabine is converted to 5'-DFUR by either
carboxyestelase or cytidine deaminase, which are localized in
liver. 5'DFUR is converted to the active form of 5-FU by thymidine
phosphorylase (dThdPase) in tumors. Capecitabine exploits the high
concentrations of dThdPase in tumor tissue as compared to healthy
tissue, leading to tumor-selective generation of 5-FU. 5-FU is
further metabolized to two active metabolites,
5-fluoro-2-deoxyuridine monophosphate (FdUMP) and 5-fluorouridine
triphosphate (FUTP), within normal and tumor cells. FdUMP inhibits
DNA synthesis by reducing normal thymidine production, while FUTP
inhibits RNA and protein synthesis by competing with uridine
triphosphate. The active moiety of capecitabine, 5-FU, is cell
cycle phase-specific (S-phase).
[0122] Studies have shown that capecitabine exhibits superior
response as compared to 5-FU/LV which is administered intravenously
(i.v.). In one study, patients treated with capecitabine achieved a
significantly (P=0.0014) superior response rate of 21% compared
with 11% with 5-FU/LV. The median times to disease progression with
capecitabine and 5-FU/LV were 128 and 131 days, respectively
(nonsignificant (NS)), and the median survival times were 380 and
407 days, respectively (NS) (Hoff et al. J Clin Oncol.
19:2282-2292, 2001). In another study, patients treated with
capecitabine also achieved a significantly (P=0.027) superior
response rate of 21% compared with 14% with 5-FU/LV. The median
times to disease progression were 137 and 131 days, respectively
(NS), and the median survival times were 404 and 369 days,
respectively (NS) (Van Cutsem et al. J Clin Oncol. 19:4097-4106,
2001). Capecitabine also demonstrated an improved safety profile
compared with i.v. 5-FU/LV.
[0123] Combination therapy using capecitabine and 5-FU/LV has shown
survival benefit as compared to 5-FU/LV alone for the treatment of
colorectal cancer. Other potential combination therapies with
capecitabine for the treatment of colorectal cancer include but are
not limited to capecitabine/oxaliplatin, capecitabine/irinotecan,
capecitabine/oxliplatin/gefitimib,
capecitabine/oxaliplatin/5-FU/LV, and capecitabine/radiation
therapy. For breast cancer treatment, combination therapies include
but are not limited to capecitabine/irinotecan,
capecitabine/docetaxel (TAXOTERE.RTM.), capecitabine/paclitaxel,
and capecitabine/5'-DFUR.
[0124] Like other chemotherapeutics, there are side effects
associated with taking capecitabine. The most common side effects
with capecitabine include diarrhea, nausea, vomiting, painful
swelling of the mouth, fatigue, painful rash and swelling of the
hands or feet, low white blood cell count (which can lead to
infections), low blood platelet counts (which can lead to
bleeding), and anemia. About one of every three patients who
receives capecitabine has serious side effects, but these side
effects usually are reversible when the drug is stopped or when the
dose is lowered.
[0125] For a detailed description of the therapeutic uses of the
fluoropyrimidine analogs, including 5-FU and capecitabine see, for
example, Chabner et al., 1996.
[0126] CPT-11: Irinotecan (CPT-11) is a semi-synthetic camptothecin
analogue that inhibits topoisomerase I in the replicating cell. It
exhibits anti-tumor activity in cancer patients who fail first-line
treatment with FU/LV (Bleiberg, 1999; Stucky-Marshall, 1999).
[0127] While CPT-11 is FDA-approved as a second-line therapy for
patients with advanced colorectal cancer, the observed response
rates are on the order of only 10%-15%.
[0128] The main side effects associated with this agent include
leukopenia, anemia, nausea/vomiting, anorexia, and diarrhea. It is,
therefore, desirable to develop a modulator agent that can either
enhance the efficacy of the anti-tumor activity of CPT-11 and/or
alleviate some of the toxic side effects associated with CPT-11
treatment so that the overall quality of life and performance
status of the cancer patient is improved.
[0129] CPT-11/FU and CPT-11/FU/LV Combination: Colorectal cancer
has been reported to be the second-leading cause of death from
cancer in North America. The two drugs that are currently approved
by the FDA for the treatment of colorectal cancer are irinotecan
(CPT-11, CAMPTOSAR.RTM.) and 5-fluorouracil (FU). FU is an
antimetabolite drug, which inhibits thymidylate synthase, an enzyme
required for the synthesis of DNA. FU is commonly administered with
LV, a reduced folate that increases the affinity of FU for
thymidylate synthase. This therapy is currently used as first-line
treatment for metastatic colon cancer (Murakami K, Sakukawa, R,
Sano, M, et al., Clin Cancer Res. 5:2304-2310 (1999); van der Wilt
C. L, van Groeningen, C. J, Pinedo H. M, et al., J. Cancer Res.
Clin. Oncol. 123:595-601 (1997)). CPT-11 is a potent inhibitor of
topoisomerase I, a nuclear enzyme involved in the unwinding of DNA
during replication. CPT-11 has demonstrated antitumor activity
against metastatic colorectal cancer as second-line treatment after
the failure of FU (Kase, Y, Hayakawa, T, Togashi, Y, et al., Jpn J
Pharmacol, 75:399-405 (1997); Araki E, Ishikawa M, Iigo M, et al.,
Jpn J Cancer Res 84:697-702 (1993); Bissery M C, Vrignaud P,
Lavelle F, et al., Anti-Cancer Drugs 7:437-460 (1996); Saltz L. B,
Cox J. V, Blanke C, et al., New. Eng. J. Med. 343:905-914 (2000)).
Recently, FDA approved the triple combination use of CPT-11/FU/LV
as the firstline treatment for advanced colorectal cancer.
Unfortunately, severe diarrhea has been identified as one of the
dose-limiting toxicities among patients treated with this
combination therapy (Saltz L. B, Cox J. V, Blanke C, et al., New.
Eng. J. Med. 343:905-914 (2000); Murakami K, Sakukawa, R, Sano, M,
et al., Clin Cancer Res. 5:2304-2310 (1999); van der Wilt C. L, van
Groeningen, C. J, Pinedo H. M, et al., J. Cancer Res. Clin. Oncol.,
123:595-601 (1997).
[0130] VP-16 (etoposide): VP-16, also known as etoposide, shows
significant clinical activity against small-cell lung cancer,
testicular cancer, lymphoma and leukemia (O'Dwyer, P., et al.,
Etoposide (VP-16-213), Current Status of an Active Anti-cancer
Drug, New Engl. J. Med. 312:692-700 (1985)) and include neoplasms
seen in Hodgkin's disease, Papillomavirus and diffuse histiocytic
lymphoma.
[0131] It is believed that etoposide blocks the catalytic activity
of DNA topoisomerase II by stabilizing an enzyme-DNA complex in
which the DNA is cleaved and covalently linked to the enzyme. See
Chen, G. L., Yang, L., Rowe T. C., Halligan, B. D., Tewey, K., and
Liu, L., J. Biol. Chem., 259:13560 (1984); Ross, W., Rowe, T.,
Glisson, B., Yalowich, J., and Liu, L., Cancer Res., 44:5857
(1984); Rowe, T., Kuppfer, G., and Ross, W., Biochem. Pharmacol.,
34:2483 (1985), which are all herein specifically incorporated by
reference.
[0132] By way of background, topoisomerases are enzymes which
control the topological state of DNA. Type II topoisomerases
catalyze DNA strand passage through transient double strand breaks
in the DNA. The resulting change in the linking number of DNA
allows these enzymes to mediate DNA interconversions, such as
supercoiling and relaxation of supercoiling, catenation and
decatenation, knotting, and unknotting. See Wang, J. C., Annu Rev.
Biochem., 54:665 (1985) and Maxwell, A., and Gellert, M., Adv.
Protein Chem., 38:69 (1986), which are herein specifically
incorporated by reference.
[0133] Type II DNA topoisomerase enzymes have been shown to be
involved in a number of vital cellular processes, including DNA
replication and transcription, and chromosomal segregation. These
enzymes, therefore, are a critical target for the action of a wide
variety of anticancer drugs, including etoposide and teniposide.
The key step leading to cell death may be the capability of these
drugs to block the catalytic activity of DNA topoisomerase II, as
noted above.
[0134] Beta-L-Dioxolane-Cytidine (L-OddC):
Beta-L-dioxolane-cytidine [L-OddC] is the first nucleoside analogue
with the unnatural L configuration shown to have anticancer
activity (Grove et al., Cancer Res (1996) 56(18):4187-4191). This
compound has been shown to have a potent antitumor activity in
human prostate and hepatocellular xenograft tumor models (Grove et
al., Cancer Res (1995) 55:3008-3011). Further, L-OddC has been
shown to be effective against hyperproliferative activity in human
keratinocytes in vitro (Schwartz et al., Skin Pharmacol Appl Skin
Physiol (1998) 11(4-5):207-213).
[0135] This compound works by rapid translocation into cells by
both equilibrative-sensitive and -insensitive nucleoside transport
systems where it is incorporated into DNA of cells. DNA
incorporation leads to degradation of DNA into large fragments
without generation of internucleosomal laddering.
[0136] Quality of Life (QOL): Standard evaluation measures for the
success of cancer treatments include, but are not limited to,
changes in tumor mass and type as well as the rate and amount of
tumor spreading (both locally to and distant to the tumor(s) being
evaluated). One skilled in the art of chemotherapy evaluations can
also determine whether a particular treatment appears to enhance a
patient's life expectancy and quality of life (even for those
patients not responding to the usual treatments). For example,
effective treatment of gastrointestinal diseases may be determined
by several criteria, including, but not limited to, an enteritis
score (based upon a composite score of clinical symptoms such as
abdominal pain, cramping, stool guaiac and diarrhea), as well as
related endpoints such as percent chemotherapy dose delivered, days
of hospitalization, transfusions, intravenous fluid therapy,
antimotility agents, and ability to eat.
[0137] With respect to a treatment effect, the subjective symptoms
of the patient do not always coincide with the result of the test
conducted by the doctor. For example, even in the case where an
unfavorable test result is obtained, when the occurrence of urinary
incontinence and voiding are reduced, the patient believes the
treatment has worked, with the result that the quality of life
(QOL) is improved. During chemotherapy, the negative side effects
in the life of the patients, such as hair loss, reduction in
weight, loss of appetite, fatigue, diarrhea, nausea, vomiting, etc.
can be persistent and result in chronic torment, night and day,
that can be unbearable to the patients, both physically and
mentally. Thus, therapeutic effectiveness of methods of the present
invention is meant to refer not only to partial or entire relief
from the pain or reduction in tumor growth or cancer regression,
but relief as a consequence of reduced or eliminated side-effects
traditionally associated with treatment, with the overall result
being an enhanced quality of life.
[0138] Baseline evaluations can be entered as part of the treatment
protocol whereby various criteria are measured and correlated with
QOL. Further, patients can report on a patient diary events such as
feeling "fair" or experiencing "moderate" pain. These measures are
then used during and after treatment to evaluate whether the
patient feels that the quality of life has improved.
[0139] PHY906
[0140] Introduction: PHY906 is a traditional Chinese botanical
formulation composed of four herbs each of which is selected from
one of four herb groups. The four herb groups are commonly known as
Scutellaria, sometimes known as Scute, Licorice, Peony Alba and
Ziziphi Fruit (Table 2). Thus, one plant species is chosen from
each one of the four plant groups provided in Table 2 in order to
produce the desired herbal compositions of the present invention.
While particular combinations of the listed plant species are
provided as examples of preferred PHY906 formulations, the
compositions and methods of this invention encompass any
combination of four plant species wherein a plant species is
selected from each one of the four groups in Table 2. This
invention encompasses any such combination of such herbs which have
at least one of the biological activities or desired effects
ascribed to PHY906 as described herein.
TABLE-US-00002 TABLE 2 Examples of Particular Species of Four
Genera Which Can be Used to Make PHY906. Common English Name of TCM
Herbal Group Scutellaria Licorice Peony Alba Ziziphi Fruit Anemone
rivularis Buch.- Abrus mollis Paeonia delavayi Franch. Ziziphus
jujuba Ham. ex DC. Hance var. lutea (Delavay ex Mill. Franch.)
Finet et Gagnep. Thalictrum omelense Glycyrrhiza Paeonia lactiflora
Pall. Ziziphus jujuba W. T. Wang et aspera Pall. Mill. var. S. H.
Wang inermis Mahonia bealei (Fort.) Glycyrrhiza Paeonia mairei
Levi. Carr. eurycarpa P. C. Li Nandina domestica Glycyrrhiza
Paeonia obovata Maxim. Thunb. glabra L. var. willmottiae (Stapi)
Stern Scutellaria amoena C. H. Glycyrrhiza Daphne papyracea Wall.
Wright inflata Bat. ex Steud. Scutellaria amoena C. H. Glycyrrhiza
Cynanchum otophyllum Wright var. cinerea squamulosa Schneid.
Hand.-Mazz. Franch. Scutellaria baicalensis Glycyrrhiza Codonopsis
lanceolara Georgi uralensis Fisch. Sieb. et Zucc. Trautv.
Scutellaria baicalensis Phlomis Georgi var. albiflora betonicoides
K. Onuma Diels Scutellaria chungtienensis C. Y. Wu Scutellaria
hypericifolia Levl Scutellaria likiangensia Diels Scutellaria
obtusifolia Hemsl. var. trinervata (Vant.) C. Y. Wu et H. W. Li
Scutellaria regeliana Nakai Scutellaria regeliana Nakai var.
ikonnikovii (Juz.) C. Y. Wu et H. W. Li Scutellaria rehderiana
Diels
[0141] This herbal formula has been long used in Asia to treat a
variety of ailments such as cardiac distention, abdominal spasms,
fever, headache, vomiting, retching, thirst and mucous-like stool
(Hani Oka and Taki No, 1998). A preferred formulation of PHY906 is
provided in Table 3.
TABLE-US-00003 TABLE 3 Herbal Ingredients of TCM Formula PHY906.
Scientific Common Name Percentage Name Traditional Use Scutellaria
33.3 Scute Baical Used to reduce capillary perme- baicalensis
Skullcap ability; to reduce inflammation: Root to treat enteritis
and dysentery; increase the secretion of bile to treat jaundice; to
relieve muscle spasms to treat coughing; to expel parasites.
Glycyrrhiza 22.2 Licorice Used to moisten the lungs and uralensis
Root stop coughs; to relax spasm and stop pain; to moderate the
action of herbs: to reduce fire and release toxins. Ziziphus 22.2
Date Has diuretic and strengthening jujuba effects Paeonia 22.2
White Peony Used to suppress and soothe lactiflora Root pain: to
soothe ligaments and purify the blood
[0142] An alternative formulation of PHY906 has the herbs
Scutellaria, Glycyrrhiza, Ziziphus, and Paeonia in the following
relative proportions: 4/14:3/14:4/14:3/14, respectively.
[0143] While specific ratios of the herbs of PHY906 are provided as
examples, the compositions and methods of this invention encompass
any ratios of the four herbal components which have the desired
biological activity as described herein.
[0144] Currently, both gelatin capsules and granule pouches of
PHY906 are produced by Sun Ten Laboratories, Inc., in Irvine,
Calif. (a sister company of Sun Ten Pharmaceutical Co. Ltd. in
Taiwan) using the formulation provided in Table 3. This formulation
of PHY906 has been distributed and sold as a dietary supplement
since 1983 by Brion Herbs Corporation (12020 B Centralia Road,
Hawaiian Garden, Calif., 90716).
[0145] Production: A brief review of a process which can be used
for producing PHY906 is provided. First, the proper ratios of the
ingredients of the herbal raw materials are placed in a jacketed
reactor and extracted with water at an elevated constant
temperature with mixing. The ratios are set forth in the
Manufacturing Instruction reproduced from Master Formula Record.
The solid materials are then separated from the liquid with a
120-mesh screen. The filtrate is collected and then concentrated by
evaporating the water under reduced pressure. The concentrated
liquor is spray dried at an elevated temperature to yield dry
powder which is then processed to yield granulated powder. This
bulk substance is then formulated into the desired dosage form.
[0146] Process controls are utilized to ensure the uniformity and
integrity of the product. Such process controls include, but are
not limited to, checking the volume of the process liquor, HPLC
determinations to establish Chemical Fingerprintings to verify
identity of the raw materials, and inspections and tests of
intermediate and final products. Accepted Quality Level (AQL)
Limits are established for each conducted analysis and for each
step of the manufacturing and control of production.
[0147] All of the components used in the production process are
assigned a specific lot number in the Production Instruction
Record. Quality control records are reviewed before a batch is
released.
[0148] Purified marker substances are used for identification and
quality control of the raw materials as well as the herbal
substances. Table 4 lists the marker substances of each raw
material used in the preparation of PHY906 herbal substance.
TABLE-US-00004 TABLE 4 Marker Substances for Herbal Ingredients of
PHY906. Origin of Herb Herb Producing Place Marker Substance
Scutellaria baicalensis Shang Xi Province, China Baicalin Georgi.
Glycyrrhiza uralensis Inner Mongolia, China Glycyrrhizin Fisch.
Ziziphus jujuba Mill. Hebei/Shangtong Chelidonic Acid Province,
China Paeonia lactiflora Pall. An Hwei Province, China
Paeoniflorin
[0149] Hepatocellular carcinoma (HCC) is currently treated by
surgical procedures and chemotherapy. Surgical removal and
postoperative therapies may improve the outlook for some patients.
Unfortunately, the vast majority of patients with hepatocellular
carcinoma will have unresectable cancers. In late 2007, sorafenib
became the first FDA-approved chemotherapeutic agent for HCC.
Published clinical studies indicate significant anti-tumor effects
(20, 21). Oral multikinase inhibitor sorafenib (BAY 43-9006) has a
dual-action on Raf kinase and vascular endothelial growth factor.
Sorafenib prevents tumor growth by combining inhibition in tumor
cell proliferation and tumor angiogenesis. Preclinical studies
suggest that sorafenib may offer therapeutic benefits in HCC by
blocking the Raf-1 signal transduction pathway.
[0150] Colorectal cancer has been reported to be the third most
common cause of death from cancer in the United States (22).
Recently, the FDA approved the triple combination use of
Oxaliplatin/5-FU/LV as the first-line treatment for patients with
advanced colorectal cancer. oxaliplatin is a synthesized
diaminocyclohexane platinum compound, which like cisplatin, causes
platinum-DNA adduct formation and destroys the integrity of DNA
(23). Other types of chemotherapeutic agents such as 5-FU and
CPT-11 are common chemotherapeutic agents used in the treatment of
colorectal cancer. Unfortunately, severe diarrhea has been
identified as one of the dose-limiting toxicities among patients
treated with chemotherapy.
[0151] Our studies showed that PHY906, an herbal composition, not
only reduced chemotherapy-induced toxicities, including body weight
loss and mortality, but it also enhanced the antitumor efficacy of
a broad-spectrum of anticancer agents, including, but not limited
to, CPT-11, 5-FU, CPT-11/5-FU/LV, VP-16, L-OddC and
oxaliplatin/5-FU/LV in colorectal cancer; sorafenib, capecitabine,
thalidomide, and CPT-11 in liver cancer; and capecitabine,
oxaliplatin, gemcitabine and gemcitabine/oxaliplatin in pancreatic
cancer in vivo animal models. The positive results from these
preclinical studies demonstrate that PHY906 can be used as an
adjuvant for a broad-spectrum of different types of
chemotherapeutic agents in anti-cancer therapy. These
chemotherapeutic agents include, but are not limit to, capecitabine
and sorafenib. The cancers include, but are not limited to,
colorectal. liver, and pancreatic cancers. The methods of the
present invention can be used to improve the quality of life of
patients including mammals undergoing chemotherapy. Specifically,
this invention relates to the dosing and scheduling of PHY906 in
potentiating the therapeutic index of a broad-spectrum of cancer
chemotherapeutic agents by the herbal composition PHY906.
[0152] The herbal compositions of the present invention are
particularly useful with cancer chemotherapies, such as, but not
limited to, treatment with irinotecan (CPT-11, CAMPTOSAR.RTM.),
5-fluorouracil (FU or 5-FU), leucovorin (LV), VP-16,
beta-L-Dioxolane-cytidine (L-OddC), capecitabine, gemicitabine,
oxaliplatin, doxorubicin, thalidomide, and combinations thereof,
such as but not limited to FU/LV, CPT-11/FU/LV, oxaliplatin/FU/LV,
and gemcitabine/oxaliplatin. The present invention provides
compositions comprising PHY906 and one or more chemotherapeutic
agents such as but not limited to CPT-11, 5-FU, VP-16, L-OddC,
capecitabine, gemicitabine, oxaliplatin, and combinations thereof
such as, but not limited to, FU/LV, CPT-11/FU/LV, and
oxaliplatin/FU/LV. The present invention also provides compositions
comprising PHY906 and one or more analogs and derivatives of
chemotherapeutic agents such as CPT-11, 5-FU, VP-16, L-OddC, LV,
capecitabine, gemicitabine, doxorubicin, thalidomide, and
oxaliplatin.
[0153] In one embodiment, the present invention provides a
composition comprising a pharmaceutically acceptable carrier,
materials or chemicals from a plant species of each of the
following genera of herbs: Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia, and one or more chemotherapeutic compounds. In another
embodiment, the materials or chemicals from a plant species is in a
form of a herbal composition comprising Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia. In yet another embodiment, the herbal
composition consists essentially of Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia.
[0154] In one embodiment, the plant species comprise Scutellaria
baicalensis, Glycyrrhiza uralensis, Ziziphus jujuba, and Paeonia
lactiflora. In another embodiment of the invention one or more
chemotherapeutic compounds are cancer chemotherapeutics. In one
embodiment of the invention, the cancer chemotherapeutics are
selected from the group consisting of capecitabine, sorafenib, and
a combination thereof.
[0155] In one embodiment of the invention, a therapeutically
effective amount of a composition comprising a pharmaceutically
acceptable carrier, materials or chemicals from a plant species of
each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia, and one or more chemotherapeutic compounds is
used to treat a disease in a mammal in need thereof. In another
embodiment, the materials or chemicals from a plant species is in a
form of an herbal composition comprising Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia. In yet another embodiment, the herbal
composition consists essentially of Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia.
[0156] In one embodiment, the present invention provides a method
of treating a disease in a mammal. The method comprises
administering to the mammal in need thereof a therapeutically
effective amount of a composition comprising a pharmaceutically
acceptable carrier, materials or chemicals from a plant species of
each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia, and one or more chemotherapeutic compounds.
In another embodiment, the materials or chemicals from a plant
species is in a form of a herbal composition comprising
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia. In yet another
embodiment, the herbal composition consists essentially of
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia.
[0157] In one embodiment, the present invention provides a method
of relieving the side effects of a chemotherapeutic compound in a
mammal. The method comprises administering to the mammal in need
thereof a composition comprising a pharmaceutically acceptable
carrier, materials or chemicals from a plant species of each of the
following genera of herbs: Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia, and one or more chemotherapeutic compounds.
[0158] In one embodiment of the invention, a composition comprising
a pharmaceutically acceptable carrier, materials or chemicals from
a plant species of each of the following genera of herbs:
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia, and one or more
chemotherapeutic compounds is administered to a mammal to enhance
the therapeutic effectiveness of the chemotherapeutic compound. In
another embodiment, the materials or chemicals from a plant species
is in a form of an herbal composition comprising Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia. In yet another embodiment, the
herbal composition consists essentially of Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia.
[0159] In one embodiment of the invention, a composition comprising
a pharmaceutically acceptable carrier, materials or chemicals from
a plant species of each of the following genera of herbs:
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia, and one or more
chemotherapeutic compounds is administered to a mammal to enhance
the antitumor activity of a chemotherapeutic compound. In another
embodiment, the materials or chemicals from a plant species is in a
form of a herbal composition comprising Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia. In yet another embodiment the herbal
composition consists essentially of Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia.
[0160] In one embodiment of the invention, a therapeutically
effective amount of a composition comprising a pharmaceutically
acceptable carrier, materials or chemicals from a plant species of
each of the following genera of herbs: Scutellaria, Glycyrrhiza,
Ziziphus and Paeonia, and one or more chemotherapeutic compounds is
administered to a mammal to treat tumors. In another embodiment,
the materials or chemicals from a plant species is in a form of an
herbal composition comprising Scutellaria, Glycyrrhiza, Ziziphus
and Paeonia. In yet another embodiment, the herbal composition
consists essentially of Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia.
[0161] In one embodiment of the invention, a composition comprising
a pharmaceutically acceptable carrier, materials or chemicals from
a plant species of each of the following genera of herbs:
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia, and one or more
chemotherapeutic compounds is administered to a mammal to inhibit
the growth of tumors in mammals. In another embodiment, the
materials or chemicals from a plant species is in a form of an
herbal composition comprising Scutellaria, Glycyrrhiza, Ziziphus
and Paeonia. In yet another embodiment, the herbal composition
consists essentially of Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia.
[0162] In one embodiment of the invention, a composition comprising
a pharmaceutically acceptable carrier, materials or chemicals from
a plant species of each of the following genera of herbs:
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia, and one or more
chemotherapeutic compounds is used to inhibit the growth of tumors.
In another embodiment, the materials or chemicals from a plant
species is in a form of an herbal composition comprising
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia. In yet another
embodiment, the herbal composition consists essentially of
Scutellaria, Glycyrrhiza, Ziziphus and Paeonia. In one embodiment,
the tumors are present in a mammal or in the form of cells in
vitro.
[0163] In one embodiment, the present invention provides a method
of improving the quality of life of a mammal undergoing
chemotherapy. The method comprises administering a therapeutically
effective amount of one or more chemotherapeutic compounds and a
composition comprising: i) a pharmaceutically acceptable carrier;
ii) materials or chemicals from a plant species of each of the
following genera of herbs: Scutellaria, Glycyrrhiza, Ziziphus and
Paeonia; and iii) one or more chemotherapeutic compounds. In
another embodiment, the materials or chemicals from a plant species
is in a form of an herbal composition comprising Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia. In yet another embodiment, the
herbal composition consists essentially of Scutellaria,
Glycyrrhiza, Ziziphus and Paeonia. Preferably, the mammal as
referenced herein is a human.
[0164] The above-referenced chemotherapeutic agents or compounds,
genera of herbs, and other terms and phrases have been described
and defined with details in the following patent applications and
patent: U.S. patent application Ser. No. 09/522,055, filed Mar. 9,
2000; International Patent Application No. PCT/US2001/007353, filed
Mar. 8, 2001; U.S. patent application Ser. No. 10/220,876, filed
Dec. 30, 2002, and issued as U.S. Pat. No. 7,025,993 on Apr. 11,
2006; U.S. Provisional Patent Application Ser. No. 60/625,943,
filed Nov. 9, 2004; U.S. patent application Ser. No. 11/100,433,
filed Apr. 7, 2005, and issued as U.S. Pat. No. 7,534,455 on May
19, 2009; and International Patent Application No.
PCT/US2005/040605, filed Nov. 9, 2005, the content of which are
herein incorporated by reference in their entirety for all
purposes.
[0165] Pharmaceutical Formulations of PHY906
[0166] The compositions of the present invention can be
administered via parenteral, subcutaneous, intravenous,
intramuscular, intraperitoneal, transdermal, or buccal routes.
Alternatively, or concurrently, administration may be by the oral
route. The dosage administered will be dependent upon the age,
health, and weight of the recipient, the type of concurrent
treatment, if any, the frequency of treatment, and the nature of
the effect desired.
[0167] The pharmaceutical formulation for systemic administration
according to the invention may be formulated for enteral,
parenteral, or topical administration. Indeed, all three types of
formulations may be used simultaneously to achieve systemic
administration of the active ingredient.
[0168] The present invention further provides compositions
containing one or more agents which treat various types of cancer
and/or modulate hematopoietic activity, such as the immunodulation
of tuberculosis (T.B.), natural killer cells (NK), monocytes, and
dendritic cells.
[0169] While individual needs vary, determination of optimal ranges
of effective amounts of each component is within the skill of the
art.
[0170] In addition to the pharmacologically active agent, the
compositions of the present invention may contain suitable
pharmaceutically acceptable carriers comprising excipients and
auxiliaries\which facilitate processing of the active compounds
into preparations which can be used pharmaceutically for delivery
to the site of action.
[0171] PHY906 can be used in the form of a medicinal preparation,
for example, in solid, semi-solid or liquid form which contains
PHY906, as an active ingredient, in admixture with an organic or
inorganic carrier or excipient suitable for external, enteral, or
parenteral applications. The active ingredient may be compounded,
for example, with the usual non-toxic pharmaceutically acceptable
carriers for tablets, pellets, capsules, suppositories, solutions,
emulsions, suspensions, and any other form suitable for use.
Formulations of the present invention encompass those which include
talc, water, glucose, lactose, gum acacia, gelatin, mannitol,
starch paste, magnesium trisilicate, com starch, keratin, colloidal
silica, potato starch, urea, and other carriers suitable for use in
manufacturing preparations, in solid, semisolid or liquid form and
in addition auxiliary, stabilizing, thickening and coloring agents,
and perfumes may be used.
[0172] For preparing solid compositions such as tablets or
capsules, PHY906 is mixed with a pharmaceutical carrier (e.g.,
conventional tableting ingredients such as corn starch, lactose,
sucrose, sorbitol, talc, stearic acid, magnesium stearate,
dicalcium phosphate or gums) and other pharmaceutical diluents
(e.g., water) to form a solid preformulation composition containing
a substantially homogeneous mixture of PHY906, or a non-toxic
pharmaceutically acceptable salt thereof. When referring to the
preformulation compositions as substantially homogenous, it is
meant that the active ingredients are dispersed evenly throughout
the composition so that the composition may he readily subdivided
into equally effective unit dosage forms such as tablets, pills and
capsules. This solid preformulation composition is then subdivided
into unit dosage forms of the type described above containing an
effective amount of the composition of the present invention,
preferably in capsules.
[0173] The tablets or pills containing PHY906 can be coated or
otherwise compounded to provide a dosage form affording the
advantage of prolonged action. For example, the tablet or pill can
comprise an inner dosage an outer dosage component, the latter
being in the form of an envelope over the former. The two
components can he separated by an enteric layer which serves to
resist disintegration in the stomach and permits the inner
component to pass intact into the duodenum or to be delayed in
release. A variety of materials can he used for such enteric layers
or coatings such materials including a number of polymeric acids
and mixtures of polymeric acids with such materials as shellac,
cetyl alcohol, and cellulose acetate.
[0174] The liquid forms, in which PHY906 may be incorporated for
administration orally or by injection, include aqueous solution,
suitably flavored syrups, aqueous or oil suspensions, and flavored
emulsions with edible oils such as cottonseed oil, sesame oil,
coconut oil, or peanut oil as well as elixirs and similar
pharmaceutical vehicles. Suitable dispersing or suspending agents
for aqueous suspensions include synthetic natural gums, such as
tragacanth, acacia, alginate, dextran, sodium carboxymethyl
cellulose, methylcellulose, polyvinylpyrrolidone or gelatin.
[0175] Liquid preparations for oral administration may take the
form of, for example, solutions, syrups or suspensions, or they may
be presented as a dry product for reconstitution with water or
other suitable vehicles before use. Such liquid preparations may he
prepared by conventional means with pharmaceutically acceptable
additives such as suspending agents (e.g., sorbitol syrup, methyl
cellulose or hydrogenated edible fats); emulsifying agents (e.g.,
lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily
esters or ethyl alcohol); preservatives (e.g., methyl or propyl
p-hydroxybenzoates or sorbic acid); and artificial or natural
colors and/or sweeteners.
[0176] For buccal administration, the compositions of the present
invention may take the form of tablets or lozenges formulated in
conventional manners.
[0177] PHY906 may also be formulated for parenteral administration
by injection, which includes using conventional catheterization
techniques or infusion. Formulations for injection may be presented
in unit dosage form, e.g., in ampules, or in multi-dose containers,
with an added preservative. The compositions may take such forms as
suspensions, solutions or emulsions in oily or aqueous vehicles,
and may contain formulating agents such as suspending, stabilizing,
and/or dispersing agents. Alternatively, the active ingredients may
be in powder form for reconstitution with a suitable vehicle, e.g.,
sterile pyrogen-free water, before use.
[0178] Suitable formulations for parenteral administration include
aqueous solutions of the active compounds in water-soluble form,
for example, water-soluble salts. In addition, suspensions of the
active compounds as appropriate oily injection suspensions may be
administered. Suitable lipophilic solvents or vehicles include
fatty oils, for example, sesame oil, or synthetic fatty acid
esters, for example, ethyl oleate or triglycerides. Aqueous
injection suspensions may contain substances which increase the
viscosity of the suspension include, for example, sodium
carboxymethyl cellulose, sorbitol, and/or dextran. Optionally, the
suspension may also contain stabilizers. Liposomes can also be used
to encapsulate the agent for delivery into the cell.
[0179] In practicing the methods of this invention, PHY906 may be
used alone or in combination, or in combination with other
therapeutic or diagnostic agents. In certain preferred embodiments,
the compounds of this invention may be coadministered along with
other compounds typically prescribed for cancer chemotherapy
according to generally accepted medical practice. The compounds of
this invention can be utilized in vivo, ordinarily in mammals, such
as humans, sheep, horses, cattle, pigs, dogs, cats, rats and mice,
or in vitro.
[0180] Actual methods for preparing administrable compositions and
adjustments necessary for administration to subjects will be known
or apparent to those skilled in the art and are described in more
detail in, for example, Remington's Pharmaceutical Science, 17th
Ed., Mack Publishing Company, Easton, Pa. (1985), which is
incorporated herein by reference.
[0181] Methods of Using PHY906
[0182] The present invention provides methods of using PHY906 in
combination with therapeutics for the treatment of various
diseases, conditions, or disorders.
[0183] Specifically, the present invention provides methods of
using PHY906 in combination with chemotherapeutic agents for the
treatment of diseases, conditions, or disorders. Preferably, the
present invention provides a method of treating cancer comprising
administering one or more cancer chemotherapeutic agents in
combination with PHY906 to a patient. The chemotherapeutic agents
include but are not limited to CPT-11, 5-FU/LV, VP-16, L-OddC,
capecitabine, gemcitabine, oxaliplatin, thalidomide, doxorubicin,
and combinations thereof. More preferably, the present invention
provides a method of treating colorectal cancer, pancreatic cancer,
or hepatocellular carcinoma comprising administering CPT-11/FU/LV,
oxaliplatin/FU/LV, or oxaliplatin/gemcitabine in combination with
PHY906.
[0184] In one aspect of the invention, PHY906 is administered to
cell lines, for example cancer or carcinoma cell lines and HIV cell
lines, to evaluate the toxicity of PHY906 on different cell lines.
Preferably, the cancer or carcinoma cell lines include, but are not
limited to Jurkat, KB, HepG2, HepG1.6, T-cell lymphoma (CEM), Colon
26, Colon 38, HCT116, PANC 01, PANC 02, HPAC, and the HIV cell
lines include, but are not limited to, H9 cells and MT-2 cells.
[0185] In another aspect of the invention, PHY906 in combination
with one or more chemotherapeutic or antiviral agents is
administered to an animal to determine whether PHY906 is effective
in increasing the therapeutic index of the agent and the quality of
life of the animal undergoing chemotherapeutic or antiviral
therapy. Preferably, the animal is a mammal. More preferably, the
mammal is a human.
[0186] The animal could be an animal model for a specific cancer or
viral disease. Also, the animal could have a deficient immune
system. Such animal models are well-known in the art.
Naturally-occurring immunodeficient mice have been used to study
the immune system, cancer, and infectious diseases, including
acquired immune deficiency syndrome or AIDS. For example, the nude
(NU) mouse is athymic, so T cell differentiation and maturation
cannot occur. Nude mice have served for many years as hosts for
xenografts, especially human tumors and the testing of anti-cancer
drugs. The severe combined immunodeficiency syndrome (SCID) mouse
appears to defectively rearrange both TCR (T cell receptor) and
immunoglobulin genes and displays a severe immunodeficiency. The
beige (BG) mouse carries a defect in functional natural killer
cells, whereas the X-linked immunodeficient (XID) mouse has a
defect in the production of B cells. In addition, crosses have been
made among various strains to generate lines with more
comprehensive immunodeficient phenotypes (e.g., BG/NU and
BG/NU/XID).
[0187] Other laboratory animals which possess little or no immune
system of their own, or which have been treated with drugs or
radiation, or produced through traditional genetic development or
genetic engineering to have either a suppressed immune system, a
weakened immune system or a modified immune system, or no immune
system at all, such as, e.g. SCID horses and other SCID animals and
potentially even AIDS infected animals in which AIDS has been
arrested after destruction or inactivation of the animal's immune
system may be considered as laboratory animal candidates for use in
the present invention (Perryman L. E., McGuire, T. C., Torbeck, R.
L., and Magnuson, N. So, Clin. Immunol. Immunopath., 23(1):1-9,
1982).
[0188] In a preferred embodiment, the animals treated with one or
more chemotherapeutic or antiviral agents in combination with
PHY906 are evaluated for weight loss and survival rate and compared
to control animals which are only administered the one or more
chemotherapeutic or antiviral agents. The effect of PHY906 on the
antitumor or antiviral activity could also be evaluated to
determine the efficacy of PHY906.
[0189] Formulations for Combination Therapy
[0190] The present invention provides combination therapy
comprising a composition containing one or more compounds and
PHY906 for the treatment of cancer, specifically colorectal cancer,
pancreatic cancer, and hepatocellular carcinoma. Preferably, the
compounds are chemotherapeutic agents such as CPT-11, 5-FU, LV,
VP-16, L-OddC, capecitabine, gemcitabine, thalidomide, doxorubicin,
and oxaliplatin. The compounds could also be antiviral agents such
as AZT, DDI, 3TC, and D4T. The combination therapy may administer
the compounds together with PHY906 as a composition or administer
the compounds separately from the administration of PHY906. Therapy
may be performed with the composition of the present invention
alone or in conjunction with another therapy (e.g., surgery,
radiation, biologic therapy).
[0191] The administration dosage and frequency of each component of
the composition can be controlled independently. For example, one
component may be administered orally three times per day, while the
second component may be administered intramuscularly once per day.
The compounds and PHY906 may also be formulated together such that
one administration delivers both components. Formulations and
dosages are described below.
[0192] Formulation of Pharmaceutical Compositions: The
administration of each chemotherapeutic agent and PHY906 of the
composition may be by any suitable means that results in a
concentration of the compound that, combined with the other
component, is specifically anti-neoplastic upon reaching the target
region. The chemotherapeutic agent may be contained in any
appropriate amount in any suitable carrier substance, and is
generally present in an amount of 1-95% by weight of the total
weight of the composition. The composition may be provided in a
dosage form that is suitable for the oral, parenteral (e.g.,
intravenously, intramuscularly), rectal, cutaneous, nasal, vaginal,
inhalent, skin (patch), or ocular administration route. Thus, the
composition may be in the form of, e.g., tablets, capsules, pills,
powders, granulates, suspensions, emulsions, solutions, gels
(including hydrogels), pastes, ointments, creams, plasters,
drenches, delivery devices, suppositories, enemas, injectables,
implants, sprays, or aerosols. The pharmaceutical compositions may
be formulated according to conventional pharmaceutical practice
(see, e.g., Remington: The Science and Practice of Pharmacy. (19th
ed.) ed. A. R. Gennaro, 1995. Mack Publishing Company, Easton, Pa.
and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick
and J. C. Boylan, 1988-1999, Marcel Dekker, New York.
[0193] Pharmaceutical compositions according to the invention may
be formulated to release the active compound substantially
immediately upon administration or at any predetermined time or
time period after administration. The latter types of compositions
are generally known as controlled release formulations, which
include (i) formulations that create a substantially constant
concentration of the drug within the body over an extended period
of time; (ii) formulations that after a predetermined lag time
create a substantially constant concentration of the drug within
the body over an extended period of time; (iii) formulations that
sustain drug action during a predetermined time period by
maintaining a relatively, constant, effective drug level in the
body with concomitant minimization of undesirable side effects
associated with fluctuations in the plasma level of the active drug
substance (sawtooth kinetic pattern); (iv) formulations that
localize drug action by, e.g., spatial placement of a controlled
release composition adjacent to or in the diseased tissue or organ;
and (v) formulations that target drug action by using carriers or
chemical derivatives to deliver the drug to a particular target
cell type.
[0194] Administration of compounds in the form of a controlled
release formulation is especially preferred in cases in which the
compound, either alone or in combination with PHY906, has (i) a
narrow therapeutic index (i.e., the difference between the plasma
concentration leading to harmful side effects or toxic reactions
and the plasma concentration leading to a therapeutic effect is
small; in general, the therapeutic index, TI, is defined as the
ratio of median lethal dose (LD.sub.50) to median effective dose
(ED.sub.50)); (ii) a narrow absorption window in the
gastro-intestinal tract; or (iii) a very short biological half-life
so that frequent dosing during a day is required in order to
sustain the plasma level at a therapeutic level.
[0195] Any of a number of strategies can be applied in order to
obtain a controlled release formulation in which the rate of
release outweighs the rate of metabolism of the compound in
question. In one example, controlled release is obtained by
appropriate selection of various formulation parameters and
ingredients, including, e.g., various types of controlled release
compositions and coatings. Thus, the drug substance is formulated
with appropriate excipients into a pharmaceutical composition that,
upon administration to the organism, releases the active substance
in a controlled manner. Examples include single or multiple unit
tablet or capsule compositions, oil solutions, suspensions,
emulsions, microcapsules, microspheres, nanoparticles, patches, and
liposomes.
[0196] Solid Dosage Forms for Oral Use: Formulations of the
composition comprising PHY906 and one or more compounds for oral
use include tablets containing the active ingredient(s) in a
mixture with non-toxic pharmaceutically acceptable excipients.
These excipients may be, for example, inert diluents or fillers
(e.g., cellulose, sorbitol, sugar, mannitol, microcrystalline
cellulose, starches including potato starch, calcium carbonate,
sodium chloride, lactose, calcium phosphate, calcium sulfate, or
sodium phosphate); granulating and disintegrating agents (e.g.,
cellulose derivatives including microcrystalline cellulose,
starches including potato starch, croscamlellose sodium, alginates,
or alginic acid); binding agents (e.g., sucrose, glucose, sorbitol,
acacia, alginic acid, sodium alginate, gelatin, starch,
pregelatinized starch, microcrystalline cellulose, magnesium
aluminum silicate, carboxymethylcellulose sodium, methylcellulose,
hydroxypropyl methylcellulose, ethylcellulose,
polyvinylpyrrolidone, or polyethylene glycol); and lubricating
agents, glidants, and antiadhesives (e.g., magnesium stearate, zinc
stearate, stearic acid, silicas, hydrogenated vegetable oils, or
talc). Other pharmaceutically acceptable excipients can be
colorants, flavoring agents, plasticizers, humectants, buffering
agents, and the like. Alternatively, the compounds may be
formulated for oral use separately from PHY906.
[0197] The tablets may be uncoated or they may be coated by known
techniques, optionally to delay disintegration and absorption in
the gastrointestinal tract and thereby providing a sustained action
over a longer period. The coating may be adapted to release the
active drug substance in a predetermined pattern (e.g., in order to
achieve a controlled release formulation) or it may be adapted not
to release the active drug substance until after passage of the
stomach (enteric coating). The coating may be a sugar coating, a
film coating (e.g., based on hydroxypropyl methylcellulose,
methylcellulose, methyl hydroxyethylcellulose,
hydroxypropylcellulose, carboxymethylcellulose, acrylate
copolymers, polyethylene glycols and/or polyvinylpyrrolidone), or
an enteric coating (e.g., based on methacrylic acid copolymer,
cellulose acetate phthalate, hydroxypropyl methylcellulose
phthalate, hydroxypropyl methylcellulose acetate succinate,
polyvinyl acetate phthalate, shellac, and/or ethylcellulose).
Furthermore, a time delay material such as, e.g., glyceryl
monostearate or glyceryl distearate may be employed.
[0198] The solid tablet compositions may include a coating adapted
to protect the composition from unwanted chemical changes (e.g.,
chemical degradation prior to the release of the active drug
substance). The coating may be applied on the solid dosage form in
a similar manner as that described in Encyclopedia of
Pharmaceutical Technology, supra.
[0199] The compounds and PHY906 may be mixed together in the
tablet, or may be partitioned. In one example, the first drug is
contained on the inside of the tablet, and the second drug is on
the outside, such that a substantial portion of the second drug is
released prior to the release of the first drug.
[0200] Formulations for oral use may also be presented as chewing
tablets, or as hard gelatin capsules wherein the active ingredient
is mixed with an inert solid diluent (e.g., potato starch, lactose,
microcrystalline cellulose, calcium carbonate, calcium phosphate or
kaolin) or as soft gelatin capsules wherein the active ingredient
is mixed with water or an oil medium, for example, peanut oil,
liquid paraffin, or olive oil. Powders and granulates may be
prepared using the ingredients mentioned above under tablets and
capsules in a conventional manner using, e.g., a mixer, a fluid bed
apparatus or a spray drying equipment.
[0201] Controlled Release Oral Dosage Forms: Controlled release
compositions containing the compounds alone or in combination with
PHY906 for oral use may, e.g., be constructed to release the active
drug substance by controlling the dissolution and/or the diffusion
of the active drug substance.
[0202] Dissolution or diffusion controlled release can be achieved
by appropriate coating of a tablet, capsule, pellet, or granulate
formulation of compounds, or by incorporating the compound in
question into an appropriate matrix. A controlled release coating
may include one or more of the coating substances mentioned above
and/or, e.g., shellac, beeswax, glycowax, castor wax, carnauba wax,
stearyl alcohol, glyceryl monostearate, glyceryl distearate,
glycerol palmitostearate, ethylcellulose, acrylic resins,
dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride,
polyvinyl acetate, vinyl pyrrolidone, polyethylene,
polymethacrylate, methylmethacrylate, 2-hydroxymethacrylate,
methacrylate hydrogels, 1,3-butylene glycol, ethylene glycol
methacrylate, and/or polyethylene glycols. In a controlled release
matrix formulation, the matrix material may also include, e.g.,
hydrated methylcellulose, carnauba wax and stearyl alcohol,
carbopol 934, silicone, glyceryl tristearate, methyl
acrylate-methyl methacrylate, polyvinyl chloride, polyethylene,
and/or halogenated fluorocarbon.
[0203] A controlled release composition containing one or more of
the compounds of the claimed combinations may also be in the form
of a buoyant tablet or capsule (i.e., a tablet or capsule that,
upon oral administration, floats on top of the gastric content for
a certain period of time). A buoyant tablet formulation of the
compound(s) can be prepared by granulating a mixture of the drug(s)
with excipients and 20-75% w/w of hydrocolloids, such as
hydroxyethylcellulose, hydroxypropylcellulose and
hydroxypropylmethylcellulose. The obtained granules can then be
compressed into tablets. On contact with the gastric juice, the
tablet forms a substantially water impermeable gel barrier around
its surface. This gel barrier takes part in maintaining a density
of less than one, thereby allowing the tablet to remain buoyant in
the gastric juice.
[0204] Liquids for Oral Administration: The compounds alone or in
combination with PHY906 may be formulated as liquids for oral
administration. Powders, dispersible powders, or granules suitable
for preparation of an aqueous suspension by addition of water are
convenient dosage forms for oral administration. Formulation as a
suspension provides the active ingredient in a mixture with a
dispersing or wetting agent, suspending agent, and one or more
preservatives. Suitable dispersing or wetting agents are, for
example, naturally-occurring phosphatides (e.g., lecithin or
condensation products of ethylene oxide with a fatty acid, a long
chain aliphatic alcohol, or a partial ester derived from fatty
acids) and a hexitol or a hexitol anhydride (e.g., polyoxyethylene
stearate, polyoxyethylene sorbitol monooleate, polyoxyethylene
sorbitan monooleate, and the like). Suitable suspending agents are,
for example, sodium carboxymethylcellulose, methylcellulose, sodium
alginate, and the like.
[0205] Parenteral Compositions: The pharmaceutical composition of
the present application comprising the compounds alone or in
combination with PHY906 may also be administered parenterally by
injection, infusion or implantation (intravenous, intramuscular,
subcutaneous, or the like) in dosage forms, formulations, or via
suitable delivery devices or implants containing conventional,
non-toxic pharmaceutically acceptable carriers and adjuvants. The
formulation and preparation of such compositions is well-known to
those skilled in the art of pharmaceutical formulation. Specific
formulations can be found in Remington: The Science and Practice of
Pharmacy, supra.
[0206] Compositions for parenteral use may be presented in unit
dosage forms (e.g., in single-dose ampoules), or in vials
containing Several doses and in which a suitable preservative may
be added (see below). The composition may be in form of a solution,
a suspension, an emulsion, an infusion device, or a delivery device
for implantation, or it may be presented as a dry powder to be
reconstituted with water or another suitable vehicle before use.
Apart from the active drug(s), the composition may include suitable
parenterally acceptable carriers and/or excipients. The compounds
may be incorporated into microspheres, microcapsules,
nanoparticles, liposomes, or the like for controlled release.
Furthermore, the composition may include suspending, solubilizing,
stabilizing, pH-adjusting agents, and/or dispersing agents.
[0207] As indicated above, the pharmaceutical compositions
according to the invention may be in the form suitable for sterile
injection. To prepare such a composition, the suitable compounds
are dissolved or suspended in a parenterally acceptable liquid
vehicle. Among acceptable vehicles and solvents that may be
employed are water, water adjusted to a suitable pH by addition of
an appropriate amount of hydrochloric acid, sodium hydroxide or a
suitable buffer, 1,3-butanediol, Ringer's solution, and isotonic
sodium chloride solution. The aqueous formulation may also contain
one or more preservatives (e.g., methyl, ethyl or n-propyl
p-hydroxybenzoate). In cases where one of the compounds is only
sparingly or slightly soluble in water, a dissolution enhancing or
solubilizing agent can be added, or the solvent may include 60% w/w
of propylene glycol or the like.
[0208] Controlled Release Parenteral Compositions: Controlled
release parenteral compositions may be in form of aqueous
suspensions, microspheres, microcapsules, magnetic microspheres,
oil solutions, oil suspensions, or emulsions. Alternatively, the
compounds may be incorporated in biocompatible carriers, liposomes,
nanoparticles, implants, or infusion devices.
[0209] Materials for use in the preparation of microspheres and/or
microcapsules are, e.g., biodegradable/bioerodible polymers such as
polyglactin, poly-(isobutyl cyanoacrylate),
poly(2-hydroxyethyl-L-glutamine) and, poly(lactic acid).
Biocompatible carriers that may be used when formulating a
controlled release parenteral formulation are carbohydrates (e.g.,
dextrans), proteins (e.g., albumin), lipoproteins, or
antibodies.
[0210] Materials for use in implants can be non-biodegradable
(e.g., polydimethyl siloxane) or biodegradable (e.g.,
poly(caprolactone), poly(lactic acid), poly(glycolic acid) or
poly(ortho esters)).
[0211] Rectal Compositions: The compounds alone or in combination
with PHY906 may be formulated for rectal administration. For rectal
application, suitable dosage forms for a composition include
suppositories (emulsion or suspension type) and rectal gelatin
capsules (solutions or suspensions). In a typical suppository
formulation, the active drug(s) are combined with an appropriate
pharmaceutically acceptable suppository base such as cocoa butter,
esterified fatty acids, glycerinated gelatin, and various
water-soluble or dispersible bases like polyethylene glycols and
polyoxyethyl ene sorbitan fatty acid esters. Various additives,
enhancers, or surfactants may be incorporated.
[0212] Compositions for Inhalation: The compounds alone or in
combination with PHY906 may be formulated for inhalation. For
administration by inhalation, typical dosage forms include nasal
sprays and aerosols. In a typically nasal formulation, the active
ingredient(s) are dissolved or dispersed in a suitable vehicle. The
pharmaceutically acceptable vehicles and excipients (as well as
other pharmaceutically acceptable materials present in the
composition such as diluents, enhancers, flavoring agents, and
preservatives) are selected in accordance with conventional
pharmaceutical practice in a manner understood by the persons
skilled in the art of formulating pharmaceuticals.
[0213] Percutaneous and Topical Compositions: The compounds alone
or in combination with PHY906 may be formulated for percutaneous
and topical administration. The pharmaceutical compositions may
also be administered topically on the skin for percutaneous
absorption in dosage forms or formulations containing
conventionally non-toxic pharmaceutical acceptable carriers and
excipients including microspheres and liposomes. The formulations
include creams, ointments, lotions, liniments, gels, hydrogels,
solutions, suspensions, sticks, sprays, pastes, plasters, and other
kinds of transdermal drug delivery systems. The pharmaceutically
acceptable carriers or excipients may include emulsifying agents,
antioxidants, buffering agents, preservatives, humectants,
penetration enhancers, chelating agents, gelforming agents,
ointment bases, perfumes, and skin protective agents.
[0214] Examples of emulsifying agents are naturally occurring gums
(e.g., gum acacia or gum tragacanth) and naturally occurring
phosphatides (e.g., soybean lecithin and sorbitan monooleate
derivatives). Examples of antioxidants are butylated hydroxy
anisole (BHA), ascorbic acid and derivatives thereof, tocopherol
and derivatives thereof: butylated hydroxy anisole, and cysteine.
Examples of preservatives are parabens, such as methyl or propyl
p-hydroxybenzoate, and benzalkonium chloride. Examples of
humectants are glycerin, propylene glycol, sorbitol, and urea.
Examples of penetration enhancers are propylene glycol, DMSO,
triethanolamine. N,N-dimethylacetamide, N,N-dimethylformamide,
2-pyrrolidone and derivatives thereof, tetrahydrofurfuryl alcohol,
and AZONE.RTM.. Examples of chelating agents are sodium EDTA,
citric acid, and phosphoric acid. Examples of gel forming agents
are CARBOPOL.RTM., cellulose derivatives, bentonite, alginates,
gelatin and polyvinylpyrrolidone. Examples of ointment bases are
beeswax, paraffin, cetyl palmitate, vegetable oils, sorbitan esters
of fatty acids (Span), polyethylene glycols, and condensation
products between sorbitan esters of fatty acids and ethylene oxide
(e.g., polyoxyethylene sorbitan monooleate (Tween)).
[0215] The pharmaceutical compositions described above for topical
administration on the skin may also be used in connection with
topical administration onto or close to the part of the body that
is to be treated. The compositions may be adapted for direct
application or for introduction into relevant orifice(s) of the
body (e.g., rectal, urethral, vaginal or oral orifices). The
composition may be applied by means of special drug delivery
devices such as dressings or alternatively plasters, pads, sponges,
strips, or other forms of suitable flexible material.
[0216] Controlled Release Percutaneous and Topical Compositions:
The compounds alone or in combination with PHY906 may be formulated
for controlled release percutaneous and topical administration.
There are several approaches for providing rate control over the
release and transdermal permeation of a drug, including:
membrane-moderated systems, adhesive diffusion-controlled systems,
matrix dispersion-type systems, and microeservoir systems. A
controlled release percutaneous and/or topical composition may be
obtained by using a suitable mixture of the above-mentioned
approaches.
[0217] In a membrane-moderated system, the active drug is present
in a reservoir which is totally encapsulated in a shallow
compartment molded from a drug-impermeable laminate, such as a
metallic plastic laminate, and a rate-controlling polymeric
membrane such as a microporous or a non-porous polymeric membrane
(e.g., ethylene-vinyl acetate copolymer). The active compound is
only permitted to be released through the rate-controlling
polymeric membrane. In the drug reservoir, the active drug
substance may either be dispersed in a solid polymer matrix or
suspended in a viscous liquid medium such as silicone fluid. On the
external surface of the polymeric membrane, a thin layer of an
adhesive polymer is applied to achieve an intimate contact of the
transdermal system with the skin surface. The adhesive polymer is
preferably a hypoallergenic polymer that is compatible with the
active drug.
[0218] In an adhesive diffusion-controlled system, a reservoir of
the active drug is formed by directly dispersing the active drug in
an adhesive polymer and then spreading the adhesive containing the
active drug onto a flat sheet of substantially drug-impermeable
metallic plastic backing to form a thin drug reservoir layer. A
matrix dispersion-type system is characterized in that a reservoir
of the active drug substance is formed by substantially
homogeneously dispersing the active drug substance in a hydrophilic
or lipophilic polymer matrix and then molding the drug-containing
polymer into a disc with a substantially well-defined surface area
and thickness. The adhesive polymer is spread along the
circumference to form a strip of adhesive around the disc.
[0219] In a microreservoir system, the reservoir of the active
substance is formed by first suspending the drug solids in an
aqueous solution of water-soluble polymer, and then dispersing the
drug suspension in a lipophilic polymer to form a plurality of
microscopic spheres of drug reservoirs.
[0220] Dosages: The dosage of each compound depends on several
factors, including: the administration method, the disease to be
treated, the severity of the disease, whether the disease is to be
treated or prevented, and the age, weight, and health of the person
to be treated.
[0221] The compounds are preferably administered in an amount of
about 0.1-30 mg/kg body weight per day, and more preferably in an
amount of about 0.5-15 mg/kg body weight per day. As described
above, the compound in question may he administered orally in the
form of tablets, capsules, elixirs or syrups, or rectally in the
form of suppositories. Parenteral administration of a compound is
suitably performed in the form of saline solutions or with the
compound incorporated into liposomes. In cases where the compound
in itself is not sufficiently soluble to be dissolved, a
solubilizer such as ethanol can be applied.
[0222] In light of the foregoing general discussion, the specific
examples presented below are illustrative only and are not intended
to limit the scope of the invention. Other generic and specific
configurations will be apparent to those persons skilled in the
art.
EXAMPLES
Example 1
Materials and Methods
[0223] Drug: Sorafenib (NEXAVAR.RTM.) was purchased from Bayer
HealthCare (Leverkusen, Germany). Capecitabine (XELODA.RTM.) was
purchased from Roche Laboratories Inc. (Nutley, N.J.). The clinical
drug substance of PHY906 (PHY906-6, FDA 165542) with 10% excipient
was prepared by Sun Ten Pharmaceutical, Inc. (Taipei, Taiwan), The
PHY906 formula is composed of four herbs: Scutellaria baicalensis
Georgi, Paeonia lactiflora Pall., Ziziphus jujuba Mill and
Glycyrrhiza uralensis Fisch., with a relative weight ratio of
3:2:2:2.
[0224] Mice: Female BDF-1 mice with body weights between 16 and 20
g (4-6 weeks old) were purchased from Charles River Laboratories
(Wilmington, Mass.). Male NCr athymic nude mice with body weights
between 16 and 20 g (4-6 weeks old) were purchased from Taconic
Farms (Garmantown, N.Y.).
[0225] Preparation of Sorafenib solution: Sorafenib (200 mg/tablet)
was dissolved in 5% gum arabic as the vehicle. The final solution
contains 30 mg/ml of sorafenib.
[0226] Preparation of capecitabine solution from capecitabine
tablet: Capecitabine (150 mg/tablet) was dissolved in 40 mM citrate
buffer (pH 6.0) containing 5% gum arabic as the vehicle. The final
solution contains 36 mg/ml of capecitabine,
[0227] Preparation of herbal extract from dry powder: The
preparation of the herbal extract followed SOP#HERB-00I-PHY906.
Briefly, one gram of PHY906 dry powder, containing starch
excipient, was added to 10 ml of 80.degree. C. H.sub.2O and
incubated at 80.degree. C. for 30 minutes. The supernatant was
separated from the debris by centrifugation (12000 rpm, 10 min) at
room temperature. The concentration of PHY906 supernatant was
calculated as 90 mg/ml of PHY906 (1 g/10 ml.times.0.9), based on
the dry weight of the dry powder. The herbal extract was stored at
room temperature and used within 24 hours. Any residual precipitant
that occurred upon standing was vortexed into a suspension and used
to treat the animals.
[0228] Tumor cells: The human hepatocellular carcinoma HepG2, human
PANC-1 pancreatic cancer, and mouse Colon 38 colorectal cancer cell
lines were purchased from the American Type Culture Collection
(Rockville, Md.). The HepG2 and Colon 38 cell lines were routinely
grown in MEME media while the PANC-1 cell line was grown in DMEM
media, supplemented with 10% fetal bovine serum (FBS). The cells
were implanted into the left flank of mice. Tumor transplantation
from mice to mice was performed when the tumor reached 1500-2000
mm.sup.3.
[0229] Mouse tumor model: Tumor cells (5.times.10.sup.6 cells in
0.1 ml PBS) were transplanted subcutaneously into the left flank of
mice. After 14 days, tumor ranging in size from 300-500 mm.sup.3
was selected for drug studies. The length and width of the each
tumor was measured with sliding calipers. The tumor size was
estimated according to the following formula:
Tumor size(mm.sup.3)=length(mm).times.width(mm).sup.2/2.
[0230] The studies were conducted and the animals were maintained
at the Yale Animal Facility.
[0231] Antitumor activity of chemotherapeutic agents in the
presence or absence of PHY906: A total of 20 tumor-bearing mice
were divided into 4 groups (N=5 mice/group):
1. Vehicle
2. PHY906
[0232] 3. Chemotherapeutic agent 4. PHY906+Chemotherapeutic
agent
[0233] The first day of drug treatment was defined as day 1. PHY906
(500 mg/kg, bid) was administrated orally to the mice 30 min before
chemotherapeutic agents at the days indicated. Chemotherapeutic
agents were given either intraperitoneally or orally at the dose
and schedule indicated. The tumor size, body weight, and mortality
of the mice were monitored daily. Mice were sacrificed when the
tumor size reached 10% of body weight.
[0234] Immunohistochemistry: Formalin-fixed paraffin-embedded liver
tissue was freshly cut into slices of 4 mm. The sections were
mounted on Superfrost slides, dewaxed with xylene, and gradually
hydrated. Antigen retrieval was achieved by 0.05% citraconic
anhydride buffer (pH 7.4) at 94.degree. C. for 1 h. The primary
HIF-1.alpha., CD31 or VEGF antibodies were diluted 1:75 using
Tris-HCl buffer containing 1% BSA and 0.5% Tween-20. The primary
antibody was incubated at room temperature for 1 hour. As a
negative control, two slides were processed without primary
antibody. Detection took place by the conventional labeled
streptavidin-biotin method with alkaline phosphatase as the
reporting enzyme according to the manufacturer's instructions. DAB
(3,3'-diaminobenzidine tetrahydrochloride, purchased from
Sigma-Aldrich, St Louis, Mo.) served as chromogen. Afterwards, the
slides were briefly counterstained with hematoxylin and aqueously
mounted.
[0235] Statistical analysis and statistical power of the study
(Diggle et al. Analysis of Longitudinal Data, 2nd ed. Oxford:
Oxford Science Publications (1994): A random effects model was
employed to analyze data from similar dosing animal trials. The
PROC MIXED procedure in SAS was used to take into account the
correlation among observations collected from the same mouse.
[0236] The following model was used to analyze the longitudinal
data:
y.sub.ijk=.mu.+.alpha.t.sub.k+.beta.(I.sub.Dt.sub.k)+.gamma.(I.sub.pt.su-
b.k)+.delta.(I.sub.DI.sub.pt.sub.k)+e.sub.ijk,
where y.sub.ijk is the relative tumor size of the jth individual
with the ith group (no treatment, drug alone, PHY906 alone, and
drug+PHY906) at the kth time point, t.sub.k is the kth time point,
.alpha. is the baseline time effect (no treatment group), I.sub.D
and I.sub.p are indicator variables for having the drug treatment
and the PHY906 treatment, .beta. is the drug-specific linear time
effect, .gamma. is the PHY906-specific linear time effect, 6 is the
drug-PHY906 synergistic linear time effect, and e.sub.ijk, is the
residual (error) term. We assumed that the errors from different
individuals are independent, and errors from the same individual at
different time points follow the autoregressive model, AR(1), to
take into account the fact the observations from the same
individual within the same treatment group are more correlated, and
the responses from closer time points are more correlated within
the same individual. The PROC MIXED in SAS 8.01 was used to perform
the statistical analysis.
[0237] CPT-11 (irinotecan, CAMPTOSAR.RTM.) was purchased from
Pharmacia & Upjohn Co (Kalamazoo, Mich.). 5-Fluorouracil (5-FU
or FU), folinic acid (leucovorin, LV), loperamide, E. coli
beta-glucuronidase, methylene blue, and phenolphthalein
glucuronidate were purchased from Sigma, Co. (St. Louis, Mo.). MEME
medium was prepared at the Yale University Cancer Center, according
to the standard procedures. RPMI 1640 medium was purchased from JRH
Co. Kanamycin, pancreatin, and trypan blue were purchased from
Gibco Co. (Grand Island, N.Y.). PHY906, PHY-915, PHY-14ST, and
PHY-15ST botanical preparations were provided by Sun Ten
Pharmaceutical Inc. (Taipei, Taiwan). PHY-14ST, previously called
TJ-14ST, consists of seven herbs: Pinelliae ternata Breit.,
Scutellaria baicalensis Georgi, Coptis chinensis Franch,
Glycyrrhiza uralensis Fisch, Fructus ziziphi, Panax ginseng C. A.
Mey., and Zingiber officinale Rosc. PHY-15ST, previously called
TJ-15ST, consists of Pueraria lobata Ohwi, Coptis chinensis Franch,
Scutellaria baicalensis Georgi, and Glycyrrhiza uralensis Fisch.
PHY-915 consists of five herbs: Panax ginseng C.A. Mey., Zingiber
officinale Rosc., Atractylodes macrocephaly Koidz, Saposhnikovia
divaricata Schischk., and Citrus reticulata Blanco.
[0238] Preparation of herbal extract from dry powder: One gram of
(A) PHY906 dry powder, containing either 50% (research batch) or
10% (clinical batch) starch excipient; or (B) PHY-915, PHY-14ST, or
PHY-15ST herbal formulations, containing unknown amounts of
excipient, was added to 10 ml of 80.degree. C. H.sub.2O and
incubated at 80.degree. C. for 30 minutes. The supernatant was
separated from the debris by centrifugation (2060 g, 15 min) and
used immediately. The concentration of PHY906 supernatant is
calculated as either 50 mg/ml (from research batch) or 90 mg/ml
(from clinical batch), based on the dry weight of aqueous extract
of raw herbs. The concentrations of other herbal formulations were
considered as 100 mg/ml, based on dry weight of the powder. The
supernatant was sterilized using a 0.45 .mu.m sterile Acrodisc
filter (Gelman Sciences) for growth inhibition studies in tissue
culture.
[0239] Anti-Tumor Studies: Murine Colon 38 (1-2.times.10.sup.6
cells in 0.1 ml PBS) or human HepG2 cells (1-2.times.10.sup.6 cells
in 0.1 ml PBS) were transplanted subcutaneously into BDF1 or NCr
athymic nude mice, respectively. The length and width of the tumors
were measured daily with sliding calipers. The tumor weight was
estimated according to the following formula (Pizzorno G, Wiegand
R, Lentz S, et al., Cancer Res. 52:1660-1665 (1992)):
Tumor size(mm.sup.3)=length(mm).times.width(mm).sup.2/2.
[0240] After 10 to 14 days, mice (five animals/group) with tumor
weights ranging from 150-200 mg were selected for drug studies (Guo
X, Lerner-Tung M, Chen H X, et al., Biochem Pharmacol 49:1111-1116
(1995)). Mice were sacrificed when the tumor size reached 10% of
body weight. PHY906 was administered orally either alone or with
anti-cancer chemotherapeutic agents. The effect of PHY906 on
antitumor efficacy and the reduction of toxicity by the agents were
evaluated. CPT-11 was given intraperitoneally (i.p.) 30 min after
PHY906 administration with the selected dose. The regimen of FU/LV
combination therapy was given as follows:
(A) treatment with FU/LV alone: first dose of LV (50 mg/kg, i.p.),
one hour later the second dose of LV (50 mg/kg; i.p.), then
immediately given FU (100 mg/kg, i.p.) (B) treatment with FU/LV
plus PHY906: first dose of LV (50 mg/kg, i.p.), followed 30 minutes
later by PHY906 (500 mg/kg, orally), followed 30 min later by a
second dose of LV (50 mg/kg, i.p.), then immediately by the FU dose
(100 mg/kg, i.p.)
[0241] The CPT-11/FU/LV triple drug combination was given as
follows: Group (A) CPT-11/FU/LV only: mice were given the first
dose of LV (50 mg/kg, i.p.) one hour before administration of
CPT-11, then immediately followed by LV (50 mg/kg, i.p.) and FU
(100 mg/kg, i.p.)
[0242] Group (B) CPT-11/FU/LV plus PHY906: mice were given the
first dose of LV (50 mg/kg, i.p.) 30 min before PHY906 (500 mg/kg,
orally). Then 30 minutes after PHY906 administration, mice were
treated with CPT-11, immediately followed by LV (50 mg/kg, i.p.)
and FU (100 mg/kg, i.p.) on day 0.
[0243] The first day of treatment was defined as day 0. PHY906 was
given orally twice a day (10 am and 3 pm) for either 4 or 8 days
beginning on day 0. For the control group, mice were administrated
a vehicle, either PBS for i.p. or H.sub.2O for p.o. (oral
administration). Animals were monitored for mortality, weight loss,
and tumor size daily.
[0244] Blood Cell Counts: Blood (20 .mu.l) was taken from mice on
days 0, 3, 6, 9 and 12 with micro-capillary tubes. Blood was then
be diluted to 200 .mu.l with normal saline (0.85% Sodium Chloride).
WBC, RBC and platelets were counted by a BAKER SYSTEM 9100.RTM.
HEMATOLOGY ANALYZER (Biochem ImmunoSystems Inc., Allentown, Pa.
18103-9562).
[0245] Cell Lines and Culture Conditions: The human HepG2
(hepatocellular carcinoma), HCT 116 (colon cancer), CEM (leukemia),
and KB (oral epidermoid carcinoma) cell lines, and murine Colon 38
cell line were purchased from the American Type Culture Collection
(Rockville, Md.). The HepG2 cell line was routinely grown in MEME
media, supplemented with 10% fetal bovine serum (FBS) and 100
.mu.g/ml kanamycin. Colon 38, HCT116, KB, and CEM cell lines were
grown in RPMI 1640 media with 10% FBS and 100 .mu.g/ml kanamycin.
Cells were incubated at 37.degree. C. in a humidified atmosphere of
5% CO.sub.2: 95% air.
[0246] Cytotoxicity in human or mouse carcinoma cell lines: Cell
growth inhibition was measured using the methylene blue uptake
assay. Cancer cells (1.times.10.sup.4) were seeded into a 24-well
plate in either 1 ml of MEME medium or RPM11640 with 10% FBS and
100 .mu.g/ml kanamycin on day 0. The freshly prepared and
sterilized PHY906 extract was added to cells on day 1, at various
concentrations, and incubated at 37.degree. C. for 3 days. The
medium was then removed, and the cell layer was stained for 30 min
with 0.3 ml of 0.5% (w/v) methylene blue solution (in 50% ethanol).
The plates were washed 3 times with tap water, dried, and the cell
layer was lysed with 1 ml of 1% Sarkosyl solution (in PBS). The
lysate solution was read on an E1x800 kinetic microplate reader
(Bio-Tek Instruments, Inc.) at 595 nm.
[0247] Results
[0248] (1) Sorafenib
[0249] Effect of PHY906 in Antitumor Activity of Sorafenib in
Murine Colon 38 Bearing BDF-1 Mice
[0250] To determine whether the combinational use of PHY906 and
sorafenib in order to improve anti-tumor activity of sorafenib.
Sorafenib at dose of 30 mg/kg (BID, D1-14), in combination with a
fixed dose of PHY906 at 500 mg/kg (BID, D1-4 and 8-11), were
studied in BDF-1 mice bearing Colon 38 murine colorectal cancer. As
shown in FIG. 1, PHY906 significantly enhanced the antitumor
activity of sorafenib in Colon 38 bearing mice. Indeed, the tumor
growth was suppressed when mice received the combination of PHY906
and sorafenib.
[0251] Effect of PHY906 in (a) Antitumor Activity, (b) Blood
Vessels, (c) VEGF Level and (d) HIF-1a of Sorafenib in Human HepG2
Xenografts
[0252] PHY906 (500 mg/kg, BID, D1-4, 8-11 and 15-18) was tested on
the antitumor activity of sorafenib (30 mg/kg, BID, D1-20) in human
HepG2 bearing nude mice. As shown in FIG. 2, the combination of
sorafenib and PHY906 shrank the tumor size approximately 60% after
the first week of combination drug treatment while mice treated
with sorafenib alone did not have the shrinkage in tumor.
[0253] The immunohistochemical stainings on mouse liver indicate
that the integrity of tumor blood vessels are destroyed with the
combination treatment of PHY906 and sorafenib, as shown in FIG. 3.
The expressions of VEGF and HIF-1.alpha. are suppressed by the
combination treatment of PHY906 and sorafenib, as shown in FIGS. 4
and 5, respectively. The data also suggest that the combination
treatment of PHY906 and sorafenib affects the Fos/Juk
transcription.
[0254] (2) Capecitabine
[0255] Effect of PHY906 on the Antitumor Activity of Capecitabine
in Human Panc-1 Tumor-Bearing Nude Mice
[0256] PHY906 was previously found to potentiate the antitumor
activity of capecitabine in human HepG2 xenografts. An experiment
\vas therefore conducted to study whether PHY906 could enhance the
antitumor activity of capecitabine in human Panc-1 xenografts. A
total of 20 NCr nude mice transplanted with Panc-1 human pancreatic
carcinoma cells were divided into 4 groups (N=5 mice/group); Group
(A) vehicle control; Group (B) treated with PHY906 (500 mg/kg, bid,
day 1-4, 8-11, 15-18, 22-25 and 29-32); Group (C) treated with
capecitabine (720 mg/kg, bid, day 1-7, 15-21, and 29-32); and Group
(D) treated with PHY906 (500 mg/kg, bid, days 1-4, 8-11, 15-18,
22-25 and 29-32) plus capecitabine (720 mg/kg, bid, day 1-7, 15-21,
and 29-32). PHY906 was found to enhance the antitumor activity of
capecitabine, as shown in FIG. 6. A similar observation was found
with lower doses of capecitabine (data not shown).
Example 2
Effect of PHY906 on the Antitumor Activity of FU/LV in BDF-1 Mice
Bearing Colon 38 Tumors
[0257] FU/LV in combination shows potent antitumor activity and is
used as the firstline treatment of colorectal cancer in patients
(Goldber R. M. and Erlichman C., Oncology 12: 59-63 (1988); Saltz
L. B., Cox J. V., Blanke C, et al., New. Eng. J. Med. 343:905-914
(2000)). Therefore, experiments similar to that described above for
CPT-11 treatment were carried out with FU/LV in animals. Colon 38
tumor bearing mice were divided into four groups: Group (A);
treatment with vehicle; Group (B); treatment with PHY906 alone;
Group (C); treatment with FU/LV alone; and Group (D); treatment
with FU/LV plus PHY906. The sequence of each regimen appears in
Materials and Methods. In this set of experiments, FU/LV was given
to mice only once on day 0, whereas PHY906 was administered twice
daily for 4 consecutive days.
[0258] Changes in body weight and tumor size were monitored daily,
as shown in FIGS. 7 and 8, respectively. As depicted in FIG. 7,
little change in body weight occurred in the four groups. This
observation is in contrast to that obtained with CPT-11 treatment.
Since dose-response studies of FU/LV on body weight loss were not
performed, it is possible that the FU/LV dose administered in this
experiment was not high enough to induce toxicity and associated
body weight loss. Although body weight loss was insufficient to
demonstrate a protective effect of PHY906 on FU/LV, FIG. 8
indicates that concomitant treatment of PHY906 did not impair the
antitumor activity of FU/LV in BDF-1 mice bearing Colon 38 tumors.
The tumor growth profile of animals in Group D is slower than that
in Group C, suggesting that PHY906 may enhance the antitumor
activity of FU/LV in this animal model. In addition, the
hematological toxicity of FU/LV in treated mice concomitantly
administered PHY906 was monitored on days 4, 8, and 12. Leucopenia
or thrombocytopenia, well known side effects induced by FU/LV (van
der Wilt C. L., van Groeningen, C. J., Pinedo H. M., et al., J.
Cancer Res. Clin. Oncol. 123:595-601 (1997)), was not reversed by
PHY906 (FIG. 9).
Example 3
Pharmacokinetics of CPT-11/FU/LV in BDF-1 Mice Bearing Colon 38
Tumor in the Presence and Absence of PHY906
[0259] The pharmacokinetic data of CPT-11/FU/LV in BDF-1 mice
bearing Colon 38 tumor in the presence and absence of PHY906 are
shown in FIGS. 10-12. PHY906-6 is a clinical batch of PHY906,
containing 10% excipient (starch).
[0260] The area under the curve (AUC) of CPT-11 in plasma increases
after co-administration of PHY906 with the triple combination of
CPT-11/FU/LV. There is no significant change of CPT-11 in either
tumor or liver tissues after PHY906 co-administration.
[0261] SN-38, an active metabolite of CPT-11, remains unchanged in
plasma, liver, or tumor.
[0262] The AUCs of FU and its nucleoside/nucleotide metabolites
(FUJ+FUR+FUMP) in plasma or liver change after PHY906
co-administration with the triple combination of CPT-11/FU/LV.
Example 4
Evaluation of Toxicity of PHY906 on Different Cell Lines
[0263] Briefly, one gram of each batch of PHY906 was added with 10
ml of water (1 mg/ml). See Table 5 for the batch properties.
TABLE-US-00005 TABLE 5 Batch Properties of PHY906 Property Batch A
Batch B Origin Taiwan, Sun-Ten Taiwan, Sun-Ten Preparation method
Standard Standard
[0264] The supernatant was collected after centrifugation and
filtered through a 0.22 .mu.m filter. Five cell types were used to
test for biological effects of each batch of PHY906: a) KB cells
(ATCC cat. #CCL-17); b) HepG2 cells (ATCC cat #HB-8065); c) T-cell
lymphoma cell line (CEM cells); d) Colon 38 and e) HCT116 (ATCC cat
#CCL-247).
[0265] The carcinoma cells (1.times.10.sup.4) were seeded into a
24-well plate in either 1 ml of MEME medium or RPMI-1640 with 10%
FBS and 100 .mu.g Kanamycin on day 0. After 24 hours, the PHY906
extract was added to the cells at varying concentrations and
incubated at 37.degree. C. for 3 days. The medium was then removed
and the cells stained with 0.3 ml of 0.5% (w/v) methylene blue
solution (in 50% EtOH) for 30 min. The plates were washed 3 times
with tap water, dried, and the cell layer was lysed with 1 ml of 1%
Sarkosyl solution (in PBS). The lysate solution was read on a
E1x800 kinetic microplate reader (Bio-Tec Instruments, Inc.) at 595
nm.
[0266] Cytotoxicity studies were performed with human T-cell
lymphoma cell line (CEM). CEM cells (5.times.10.sup.4) were grown
in 1 ml RPMI 1640 medium with 20% displayed fetal bovine serum. The
PHY906 extract was added at day 0. The growth of cells was assessed
3 days post addition of PHY906. The number of cells were estimated
using a hemacytometer.
[0267] The results of the assays using the two (2) batches are
displayed in Table 6. Based on these data, PHY906 sources A and B
have relatively little toxicity for KB, CEM and HCT116 cells, while
having significantly greater cytotoxic effects against Colon 38 and
HepG2 cells (see Table 6). Similar results are shown in Example 12,
Table 9.
TABLE-US-00006 TABLE 6 Cytotoxicity of Traditional Herbal
Formulations in Different Cell Lines IC.sub.50 (mg/ml).sup.a Herbal
Colon Formulation.sup.b KB HepG2 CEM 38 HCT116 PHY906A 1.35 .+-.
0.28 .+-. 0.17 1.45 .+-. 0.45 0.08 1.3 0.52 PHY906B 1.80 .+-. 0.17
.+-. 0.12 1.28 .+-. 0.02 0.08 1.2 0.99 .sup.aBased on the dry
weight of herbal formulation. .sup.bDifferent research batch of
PHY906 containing 50% excipient.
Example 5
Determination of CPT-11 Dose on BDF-1 and Nude Mice
[0268] Animal weight loss was monitored as an indication of
toxicity caused by anticancer chemotherapy agents. The effect of
CPT-11 on weight loss in non-tumor bearing BDF-1 mice was studied
using six different dosages: 100, 200, 300, 400, 600, or 800 mg/kg
body weight to determine the maximum tolerable dose in mice. A
single bolus dose of CPT-11 was administered intraperitoneally
(i.p.) at the beginning of the study, and weight loss was monitored
daily for 12 days.
[0269] One dose of CPT-11 was administered i.p. to each mouse at
the beginning of the study, and the weight loss of the animal was
then monitored daily for 12 days.
[0270] Dosages lower than 200 mg/kg had little effect on body
weight (comparison made with control mice receiving no CPT-11
treatments). In contrast, drug doses greater than or equal to 600
mg/kg resulted in animal death on the second day after CPT-11
administration. In general, mice were able to tolerate doses up to
400 mg/kg.
[0271] The profiles of the weight loss of the surviving mice are
shown in FIG. 13. The average body weights of the mice treated with
300 mg/kg CPT-11 were significantly less than those of the mice
that received no treatment with CPT-11 until 5 days after treatment
(FIG. 13). The average body weights of these two groups of mice
were not significantly different from 5 days after treatment until
the end of the trial. Both the duration and extent of the weight
loss were sensitive to the dose of CPT-11 administered to the
animal.
[0272] Weight loss was observed immediately after CPT-11 treatment
and continued for six days in mice injected with 400 mg/kg of
CPT-11. These animals gradually recovered their original body
weight on day 12. Based on these results, either 400 mg/kg or 300
mg/kg CPT-11 was used in the BDF-1 mice model. However, nude mice
inoculated with human tumor cells were significantly more sensitive
to CPT-11 treatment than normal BDF-1 mice. The maximum tolerable
dose of CPT-11 in nude mice bearing the human HepG2 xenografts was
200 mg/kg (data not shown).
Example 6
Effect of PHY906 on CPT-11 Induced Body Weight Loss in Tumor
Bearing BDF-1 Mice
[0273] PHY906 was evaluated as a modulator of CPT-11 therapy for
toxic side effects in mice inoculated with Colon 38 tumor cells.
Based on the previous findings (Example 5), a single bolus dose of
400 mg/kg CPT-11 was selected to study the effect of PHY906 on
weight-loss associated with toxicity of CPT-11. To evaluate whether
PHY906 impairs the antitumor efficacy of CPT-11, mice were
implanted subcutaneously with Colon 38 tumor cells. Ten to 14 days
after inoculation, mice were treated with CPT-11 (400 mg/kg, i.p.)
in the absence or presence of PHY906, which was given orally twice
a day at varying doses (125 mg/kg, 250 mg/kg, and 500 mg/kg).
PHY906 treatment was continued at the dose indicated for 8
consecutive days.
[0274] FIG. 14 shows that the effect of PHY906 on weight loss in
CPT-11 treated mice is dose dependent. CPT-11 treated animals
receiving supplemental treatment with 500 mg/kg/b.i.d. of PHY906
exhibited significant improvement in maintaining body weight and
recovered their original body weight more rapidly (p<0.01).
Table 7 summarizes the statistical results. However, mice receiving
250 mg/kg/b.i.d. of PHY906 showed no difference in body weight
loss.
TABLE-US-00007 TABLE 7 Statistical Analysis of PHY906 on Weight
Loss in Tumor Bearing Mice Treated With CPT-11. CPT-11/ CPT-11/
CPT-11/ No PHY906 PHY906 PHY906 N = 5 Treatment CPT-11 500 mg/kg
250 mg/kg 125 mg/kg No -- P < 0.01 P < 0.05 P < 0.01 P
< 0.01 Treatment CPT-11 P < 0.01 -- P < 0.01 P > 0.1 P
> 0.1 CPT- P < 0.05 P < 0.01 -- P < 0.01 P < 0.01
11/PHY906 500 mg/kg CPT- P < 0.01 P > 0.1 P < 0.01 -- P
> 0.1 11/PHY906 250 mg/kg CPT- P < 0.01 P > 0.1 P <
0.01 P > 0.1 -- 11/PHY906 125 mg/kg
Example 7
Tumor Weight of Colon 38 Inoculated Mice Treated with CPT-11 and
PHY906
[0275] Mice were treated as set forth in Example 6 and evaluated
for tumor weights over a nine day period. The results demonstrate
that treatment with PHY906 neither impedes nor impairs the
antitumor efficacy of the CPT-11 (FIG. 15). In fact, the data
suggest that this herbal medicine may actually enhance CPT-11
anti-tumor activity.
[0276] These preliminary results suggest that the herbal
composition PHY906 can be used as a modulator for CPT-11
chemotherapy to significantly improve and alleviate the toxic side
effects of CPT-11 without compromising the anti-tumor efficacy of
the CPT-11.
Example 8
Effect of PHY906 on Antitumor Activity and Hematological Toxicity
of CPT-11 in BDF-1 Mice Bearing Colon 38 Tumors
[0277] Based on the results obtained in the above studies, 500
mg/kg/b.i.d. PHY906 offers the best protection for host toxicity
induced by the maximum tolerable dose of 400 mg/kg CPT-11. This set
of dosages was used for the next studies. Colon 38-bearing BDF-1
mice treated with one dose of CPT-11 (400 mg/kg, i.p.) were given
500 mg/kg/b.i.d. PHY906 orally for either 4 or 8 days. Five mice
were used in each group, and the experiment was repeated eight
times. FIG. 15 represents a typical result of all of experiments.
The antitumor activity of CPT-11, as measured by tumor size, was
not compromised by the concomitant PHY906 therapy in the animal
model. In fact, a slight reduction in tumor size occurred,
suggesting that PHY906 may potentiate the antitumor activity of
CPT-11.
[0278] Myelo-suppression is a common side effect among patients
treated with CPT-11 (Bleiberg H and Cvitkovic E., Eur J Cancer
32A(Suppl 3):S18-S23 (1996)). To evaluate whether PHY906
beneficially reverses myelo-toxicity induced by CPT-11,
hematological activity was examined in BDF-1 mice bearing Colon 38
tumors. As shown in FIG. 16, PHY906 was found to have no impact on
red blood cell count or platelet count among all regimens, and
demonstrated no protection on myelo-suppression induced by CPT-11.
With respect to the antitumor activity and hematological activity,
there was no significant difference between a 4-day or 8-day
co-treatment of PHY906 with CPT-11 (data not shown).
Example 9
Effect of PHY906 on Mortality of CPT-11 on BDF-1 Mice Bearing Colon
38 Tumors
[0279] Mice were divided into four groups with different treatment
regimens: Group (A) treatment with vehicle; Group (B) treatment
with a single dose of 400 mg/kg CPT-11 by i.p. injection; Group (C)
treatment with PHY906 (500 mg/kg/b.i.d.) alone; or Group (D)
treatment with a single dose of 400 mg/kg CPT-11 plus 500
mg/kg/b.i.d. PHY906 for 4 or 8 days. The sequence of each treatment
regimen appears in Materials and Methods. As depicted in Table 8,
of 35 tumor-bearing mice (100%) from Group A and 15 of 15 mice
(100%) from Group C survived treatment with either vehicle or
PHY906 alone for 4 or 8 days, indicating no or very low toxicity
for PHY906. In contrast, treatment with CPT-11 alone (Group B)
resulted in only 33 of 40 tumor-bearing mice (82.5%) surviving
after 8 days. However, this survival rate dramatically improved to
either 95% (19 of 20 mice) or 100% (24 of 24 mice) after receiving
4 or 8 days of PHY906 treatment in combination with CPT-11 (Group
D). This suggests that PHY906 treatment can protect mice against
mortality induced by a single dose of 400 mg/kg CPT-11.
TABLE-US-00008 TABLE 8 Effect of PHY906 on Survival of CPT-11
Treated BDF-1 Mice Bearing Colon 38 Tumor. Treatment CPT-11 PHY906
Dose Dose Total No. of Mice Medication (mg/ (mg/ Treated Survived
Survival .sup.a Regimen kg) Days kg) Days N N % A 0 0 0 0 35 35 100
B.sup.b 400 1 0 0 40 33 82.5 C 0 0 500 4 35 35 100 0 0 500 8 15 15
100 D.sup.c 400 1 500 4 20 19 95 400 1 500 8 24 24 100 .sup.a All
the animals were observed for 14 days. .sup.b7 of 40 mice with
CPT-11 treatment died on day 5 (N = 1), 6 (N = 3), 7 (N = 1) and 8
(N = 2). .sup.cWith the combination treatment of CPT-11 and PHY906
(4 days) treatment, one mouse died on day 6.
Example 10
Effect of PHY906 on Antitumor Activity of CPT-11/FU/LV in BDF-1
Mice Bearing Colon 38 Tumors
[0280] The FDA recently approved the new triple combination therapy
of CPT-11 plus FU/LV as a firstline treatment for advanced
colorectal cancer (Goldber R. M. and Erlichman C., Oncology 12:
59-63 (1988); Saltz L. B, Cox J. V, Blanke C, et al., New. Eng. J.
Med. 343:905-914 (2000)). This regimen has been proven to slow the
progress of tumor growth as well as the mortality rate. However,
severe, late-onset diarrhea is often observed in patients receiving
this triple treatment regimen. Previous experiments demonstrated
that PHY906 could increase the therapeutic index of CPT-11 in BDF-1
mice bearing Colon 38 tumors. Therefore, PHY906 was evaluated using
a similar protocol as in Example 2 for its efficacy on alleviating
the dose-limiting toxicity of triple chemotherapy. BDF-1 mice
bearing Colon 38 tumors were divided into two groups: Group (A)
treated with CPT-11/FU/LV only; Group (B) treated with CPT-11/FU/LV
plus PHY906. The doses of FU and LV used in both groups were 100
mg/kg each, because very low toxicity was observed in previous
studies at these doses. A dose-dependent study of CPT-11 was not
performed in this triple chemotherapy regimen, but either 200 mg/kg
or 300 mg/kg CPT-11 was used. The sequence of each regimen appears
in Materials and Methods. PHY906 was administered twice daily for 4
days post chemotherapy.
[0281] The results indicate that PHY906 does not impair the
antitumor efficacy of triple therapy at both 200 mg/kg and 300
mg/kg CPT-11, as shown in FIGS. 17 and 18, respectively. As
depicted in FIG. 17, using 200 mg/kg CPT-11 in triple combination
therapy, PHY906 slightly enhances tumor suppression at day 14
(p=0.045). At 300 mg/kg CPT-11, the enhancement of PHY906 in tumor
suppression was not significant at day 14 (p=0.05), but was
significant at day 21 (p=0.014), compared to groups receiving no
PHY906 treatment. This result suggests that a longer time period
may be needed to observe the enhancement of PHY906 on CPT-11/FU/LV
tumor suppression. At the dose studied, PHY906 showed a similar
beneficial effect on antitumor activity in triple combination
therapy and CPT-11 treatment.
[0282] At the doses used in the triple combination treatment,
animals showed body weight loss. However, PHY906 did not slow
weight loss during therapy, as shown in FIG. 19. In addition,
PHY906 did not affect the recovery of body weight loss.
Example 11
Effect of PHY906 on Antitumor Activity and Toxicity of CPT-11 in
Human HepG2 Tumor-Bearing Nude Mice
[0283] Results from the above Examples, specifically Examples 2, 8,
and 10, indicate that PHY906 in combination with chemotherapeutic
agents may potentiate the antitumor effects of chemotherapeutic
agents and further retard tumor growth. Based upon the known
pharmacological profiles of herbs contained in PHY906, it is
speculated that the enhancing effects may act through immunological
and/or hematological systems in normal mice. Therefore, experiments
were designed to test the hypothesis in nude mice, which are
deficient in immunological and hematological systems.
[0284] Human HepG2 tumor cells were implanted into NCr-nude mice to
test the effect of PHY906 on the antitumor activity of CPT-11.
Previous experiments showed that the maximum tolerable dose of
CPT-11 in nude mice was 200 mg/kg, which was used in this study.
CPT-11 (200 mg/kg, i.p.) was given on day 0. PHY906 was given twice
daily at 500 mg/kg starting on day 0. As shown in FIG. 20, PHY906
did not compromise the antitumor effect of CPT-11 on human HepG2
xenografts in nude mice. However, unlike the observation in BDF-1
mice, PHY906 showed no beneficial effect on preventing body weight
loss (FIG. 21) or animal death (data not shown) caused by CPT-11.
The fact that PHY906 does not protect nude mice from weight loss as
it does normal mice suggests that PHY906 exerts its effects through
hematological and immunological systems, which nude mice lack.
Example 12
Effects of Different Chinese Herbal Formulations on Antitumor
Effect of CPT-11, Body Weight Loss, and Survival in Mice
[0285] Diarrhea is one of the dose-limiting toxicities among
patients treated with cancer chemotherapeutic agents. In addition
to PHY906, other anti-diarrhea medicines were examined. These
included Chinese medicines, such as PHY-14ST, PHY-15ST, and
PHY-915, as well as loperamide, currently recommended as the
anti-diarrhea drug for CPT-11-induced late-onset diarrhea.
[0286] In addition to tumor growth inhibition and loss of body
weight, we examined survival rates in mice receiving different
herbal formulations in combination with a single bolus
administration of CPT-11. Of the several formulations examined,
PHY906 was the only one observed to enhance CPT-11 antitumor
activity (Table 8), even though certain herbs contained in PHY906
are also present in the other herbal formulations. In the mortality
study, PHY906 showed no statistical effect on animal death rates
(P=0.044). Other anti-diarrhea medicines tested, such as PHY-14ST,
PHY-15ST and loperamide, were observed to be completely ineffective
in protecting against body weight loss or enhancing the antitumor
effects of CPT-11. Surprisingly, PHY-915, was observed to decrease
CPT-11 antitumor activity (Table 9).
TABLE-US-00009 TABLE 9 Effect of Different Herbal Formulations on
CPT-11 Treated BDF-1 Mice Bearing Colon 38 Tumor. Herbal Protection
Animal Formulation from Death.sup.a (1 g.sup.c/kg, Body Weight P
Antitumor P (Death/ bid, 8 days) Loss value.sup.b,d Effect
value.sup.b,e Total) None 7/40 PHY906 Significant 0.0004
Enhancement 0.0027 0/24 PHY14ST No Change 0.1072 No Change 0.2742
1.sup.f/10 PHY15ST No Change 0.3259 No Change 0.6535 0/3 PHY915
Significant 0.0306 Decrease 0.0885 0/5 Loperamide No Change 0.9706
No Change 0.1595 3.sup.g/10.sup. .sup.aAll of the animals were
observed for 14 days. .sup.bThe p values were calculated using the
Student's paired t-test. .sup.cbased on the dry weight of
formulations which contain excipient. .sup.dCalculated on the day
that CPT-11-treated mice reached maximum body weight loss.
.sup.eCalculated on the tumor size at day 6 after initial drug
treatment. .sup.fOne mouse died on day 6. .sup.gMice died on day 3
(N = 1), 4 (N = 1) and 5 (N = 1).
Example 13
Cytotoxicity of PHY906 in Different Cell Lines
[0287] To evaluate in vitro cell models as measures of quality
assurance, the effects of two different preparations of PHY906
(PHY906A and PHY906B) on the growth inhibition of different human
tumor cell lines and mouse Colon 38 tumor cell lines were studied.
As shown in Table 10, PHY906A and PHY906B showed no significant
difference in growth inhibitory activities among the cell lines. Of
note, HepG2 cell lines were found to be more sensitive to PHY906
than other human cell lines.
TABLE-US-00010 TABLE 10 Cytotoxicity of PHY906 in Different Cell
Lines. IC.sub.50 (mg/ml).sup.a Herbal Formulation.sup.b KB HepG2
CEM HCT116 Colon 38 PHY906A 0.67 .+-. 0.14 .+-. 0.73 .+-. 0.17 0.65
0.08 .+-. 0.02 0.26 0.08 PHY906B 0.90 .+-. 0.09 .+-. 0.64 .+-. 0.01
0.6 0.07 .+-. 0.02 0.5 0.06 .sup.aBased on the dry weight of
aqueous extract of raw herbs. .sup.bDifferent research batch of
PHY906.
[0288] The results of the above experiments suggest that PHY906
reduces some host toxicities induced by treatments of CPT-11,
FU/LV, or the triple combination therapy with CPT-11/FU/LV. The
botanical drug PHY906 not only maintains, but also potentiates, the
antitumor activity of the chemotherapeutic agents tested. Indeed,
PHY906 enhances the therapeutic index of CPT-11, FU/LV and
CPT-11/FU/LV by increasing the overall antitumor activity in both
Colon 38 tumor-bearing mice and human Hep G2 xenografts in nude
mice. These observations were tested with several anticancer agents
in two different tumor models (FIGS. 8, 15, 17, 18, and 20).
Example 14
Effect of PHY906 on Tumor Growth in Colon 38 Inoculated Mice
Treated with L-OddC
[0289] PHY906 was evaluated as a modulator of L-OddC
(beta-L-Dioxolane-cytidine) therapy for tumor growth in mice
inoculated with Colon 38 tumor cells. Mice were subjected to a
subcutaneous injection of murine Colon 38 tumor cells. Seven days
after inoculation of the cancer cells, mice were treated with
L-OddC (25 mg/kg) intraperitoneally and oral administration of
PHY906 (500 mg/kg, b.i.d.). The animals were then administered only
with the same dose of PHY906 continuously for the rest of the
experiment.
[0290] As shown in FIG. 22, treatment with L-OddC demonstrates that
PHY906 neither impedes nor impairs the antitumor efficacy of the
L-OddC. In fact, the data suggest that this herbal medicine may
actually enhance L-OddC anti-tumor activity.
[0291] Thus, these results suggest that the herbal PHY906 can be
used as a modulator for L-OddC chemotherapy to significantly
improve and alleviate the toxic side effects without compromising
the anti-tumor efficacy of the L-OddC.
Example 15
Effect of PHY906 on Tumor Growth in Colon 38 Inoculated Mice
Treated with VP-16
[0292] PHY906 was evaluated as a modulator of VP-16 (etoposide, a
topoisomerase II inhibitor) therapy for tumor growth in mice
inoculated with Colon 38 tumor cells. Mice were subjected to a
subcutaneous injection of murine Colon 38 tumor cells. Seven days
after inoculation with the cancer cells, mice were treated with
VP-16 (25 mg/kg) intraperitoneally and oral administration of
PHY906 (500 mg/kg, b.i.d.). The animals were then administered only
with the same dose of PHY906 continuously for the rest of the
experiment.
[0293] As shown in FIG. 23, treatment with VP-16 demonstrates that
PHY906 neither impedes nor impairs the antitumor efficacy of the
VP-16. In fact, the data suggest that this herbal medicine may
actually enhance VP-16 anti-tumor activity.
[0294] Thus, these results suggest that the herbal PHY906 can be
used as a modulator for VP-16 chemotherapy to significantly improve
and alleviate the toxic side effects without compromising the
anti-tumor efficacy of the VP-16.
Example 16
Effect of PHY906 on Tumor Growth in Colon 38 Inoculated Mice
Treated with 5-Fluorouracil
[0295] PHY906 was evaluated as a modulator of 5-fluorouracil (FU)
therapy for tumor growth in mice inoculated with Colon 38 tumor
cells. Mice were subjected to a subcutaneous injection of murine
Colon 38 tumor cells. Seven days after inoculation of the cancer
cells, mice were treated with 5-fluorouracil at two doses (250
mg/kg, one dose on day 0, or 30 mg/kg daily dose given from day 0
to day 4) intraperitoneally, and with PHY906 (500 mg/kg, b.i.d.) by
oral administration. The animals were then administered only with
the same dose of PHY906 continuously for the rest of the
experiment.
[0296] As shown in FIGS. 24 and 25, treatment with 5-fluorouracil
demonstrates that PHY906 neither impedes nor impairs the antitumor
efficacy of the 5-fluorouracil. In fact, the data suggest that this
herbal medicine may actually enhance 5-fluorouracil anti-tumor
activity.
[0297] Thus, these results suggest that the herbal PHY906 can be
used as a modulator for 5-fluorouracil chemotherapy to
significantly improve and alleviate the toxic side effects without
compromising the anti-tumor efficacy of the 5-fluorouracil.
Example 17
Effect of PHY906 on Tumor Growth in Colon 38 Inoculated Mice
Treated with CPT-11 and Loperamide
[0298] PHY906 was evaluated as a modulator of CPT-11 therapy for
tumor growth in mice inoculated with Colon 38 tumor cells in the
presence of anti-diarrhea medication Loperamide. Mice were
subjected to a subcutaneous injection of murine Colon 38 tumor
cells. Seven days after inoculation of the cancer cells, mice were
treated with CPT-11 (400 mg/kg, i.p.), alone, in the presence of
orally administered of PHY906 (500 mg/kg, b.i.d.), or in the
presence of Loperamide (2 mg/kg, p.o., b.i.d.).
[0299] FIG. 26 compares the antitumor effect of PHY906 and
Loperamide. As shown in FIG. 26, CPT-11 in the presence of PHY906
is more effective at reducing tumor growth (as determined as a
percentage of initial tumor weight) than Loperamide.
[0300] These preliminary results suggest that the herbal PHY906 is
more effective than standard administration of Loperamide for
delayed CPT-11 induced diarrhea.
Example 18
To Determine the Minimal Effective Dose (MED) and the Optimal
Duration of PHY906 Administration When Given in Combination with
Irinotecan
[0301] Introduction: Several studies indicate that Kampo medicine,
which consists of seven herbs, is effective in preventing the
occurrence of CPT-11-induced diarrhea in animals and in reducing
the severity of CPT-11-induced diarrhea in vivo (Mori, 1998).
[0302] PHY906 has also been evaluated in an in vivo animal model
and has been shown to reduce the severity of irinotecan-induced
toxicity. Accordingly, based on a long historical experience (1500
years) demonstrating safety in humans, the promising preclinical
activity of this compound in an animal model, and the potential
activity noted for a related herbal compound in this setting, a
study can be conducted to evaluate the effect of PHY906 on the
severity of chemotherapy-induced toxicities, such as weight loss,
diarrhea, overall performance status, and quality of life, and on
the anti-tumor activity of irinotecan or other drugs in patients
with refractory advanced colorectal cancer.
[0303] This study includes patients with histologically confirmed,
FU-refractory, advanced colorectal cancer. Measurable or evaluable
disease is not required. Patients with central nervous system (CNS)
metastases are eligible provided the CNS disease has remained
stable for at least 4 weeks following completion of surgery,
chemotherapy, and/or radiation therapy.
[0304] Participants in the study will be .gtoreq.18 years of age
and will have no significant underlying medical diseases. All
patients will have a performance status of ECOG 0-2, a life
expectancy of at least 3 months, and have given informed consent.
(ECOG is an abbreviation for "Eastern Cooperative Oncology Group."
ECOG 0=patient performing normal activity; ECOG 1=patient having
minimal symptoms; ECOG 2=to patient spents <50% of time in bed;
ECOG 3=patient spents >50% time spent in bed; ECOG 4=patient is
bed bound.) Patients must have fully recovered from the effects of
any prior surgery and have not received wide-field radiation or any
chemotherapy within 4 weeks (6 weeks for nitrosoureas or mitomycin
C) of entry onto this study. An ANC (absolute neutrophil
count).gtoreq.1500 .mu.l, platelet count .gtoreq.40 ml/min, and a
total bilirubin .ltoreq.2.0 mg/dl is required for entry onto
study.
[0305] Pretreatment Evaluation: Prior to the start of treatment,
all patients will have a complete history, physical examination,
and a determination of their performance status. The laboratory
studies will include a complete blood count (CBC) with
differential, a serum albumin, electrolytes, glucose, blood urea
nitrogen (BUN), creatinine, serum calcium and magnesium, liver
function tests, prothrombin and partial thromboplastin time, and a
urinalysis.
[0306] Treatment: Irinotecan will be reconstituted from a
lyophilized powder into 2 ml of sterile water, diluted in 100 ml of
D5W (5% Dextrose in water), and administered over 90 min at a dose
of 125 mg/m.sup.2. Irinotecan chemotherapy will be administered on
a weekly schedule for 4 weeks with a 2 week rest period in the
outpatient clinic at each of the participating hospitals.
[0307] PHY906 will be taken on an empty stomach 30 min prior to
meals. On chemotherapy treatment days, the first dose will be taken
before the administration of irinotecan.
[0308] PHY906 will be administered orally three times a day before
each meal starting at an initial dose of 0.60 g (total daily dose,
1.80 gm/day). The dose of PHY906 that is presently being used by
patients in the Orient is 7.2 gm/day, and to date no adverse events
have been observed. Thus, the dose that is proposed to start out
this trial is 1/4th the usual dose of the herbal medicine. PHY906
will be given for an entire 4-week course of chemotherapy along
with irinotecan with a 2-week rest. A minimum of three patients
will be treated at this initial dose level of PHY906. Once the 3
patients have completed a full 6-week cycle, if 0 of 3 patients
experience dose-limiting toxicity (DLT), then the next higher dose
will be used for the subsequent group of 3 patients. In all
patients, pharmacokinetic studies will be performed 24 hr after the
start of the first cycle of chemotherapy.
[0309] If 1 of 3 patients experience DLT, then 3 more patients will
be treated at the same dose level. If .ltoreq.1 of the next 3
experience DLT (1 or 2 of 6 total patients), the dose will be
escalated to the next dose level except when those events occur
during the doubling scheme, when the next escalation will be to
level n+1 on the modified Fibonacci scheme (Table 11).
TABLE-US-00011 TABLE 11 Dose Escalation Schedule of PHY906. Total
Dose Dose Level Escalation (gm/day) 1 Starting 1.8 2 2 .times.
level 1 3.6 3 2 .times. level 2 7.2 Once the 7.2 gm/day dose level
is reached and no .gtoreq. grade 2 toxicity is observed at this
level "n" a modified Fibonacci escalation as shown below will be
performed. N + 1 1.5 .times. level n N + 2 1.33 .times. level n + 1
N + 3 1.25 .times. level n + 2 All subsequent levels: 25%
increments until the maximum tolerated dose is reached.
[0310] Dose Escalation Schedule of PHY906: If clinically indicated
and considered necessary by the Principal Investigators, a lower
dose level, rather than the level specified above, may be
utilized.
[0311] The rate of subject entry and escalation to the next dose
regimen will depend upon assessment of the safety profile of
patients entered at each dose level. Toxicity will be evaluated and
graded according to the NCI Clinical Trial Guidelines (CTG)
Expanded Common Toxicity Criteria.
[0312] The antiemetic schedule for this protocol will consist of
1-2 mg of granisetron admixed in 50 ml normal saline and
administered via 1/2 hr prior to chemotherapy on each treatment
day. The antiemetics (administered intravenously or orally) will be
repeated every 8 hr as needed to control nausea and/or vomiting.
Treatment will be repeated every week for 4 consecutive weeks
followed by a 2-week rest. This will constitute one cycle of
therapy.
[0313] Diarrhea that occurs during or shortly after irinotecan
infusion will be treated with atropine (0.5-1 mg) intravenously.
For diarrhea occurring .gtoreq.12 hr after irinotecan
administration, patients will be treated with loperamide 4 mg
orally at the first sign of diarrhea followed by 2 mg orally every
2 hr (4 mg orally every 4 hr at night) until there is complete
resolution of the diarrhea for at least 12 hr. If the diarrhea is
bloody, associated with fevers .gtoreq.101.6.degree. F., and
continues unabated for .gtoreq.12 hr, the patient will be admitted
to the hospital for further evaluation and treatment.
[0314] Dose Modification of Irinotecan: There will be no dose
escalation of irinotecan in this study. Dose modification for
toxicity will be made as recommended in the package insert provided
by the manufacture (Table 12).
TABLE-US-00012 TABLE 12 Recommended Dose Modifications for the
Weekly and Once-Every 3-Week Schedule. A new course of therapy
should not begin until the granulocyte count has recovered to
.gtoreq.1500/mm.sup.3, and the platelet count has recovered to
.gtoreq.100,000/mm.sup.3, and treatment-related diarrhea is fully
resolved. Treatment should be delayed 1 to 2 weeks to allow for
recovery from treatment-related toxicities. If the patient has not
recovered after a 2 week delay, consideration Should be given to
discontinuing CAMPTOSAR .RTM.. (irinotecan, CPT-11). At the Start
of the Next Courses of Therapy (After Adequate Recovery), Compared
with the Weekly Toxicity Starting Dose in the Previous Course.sup.a
NCI Grade.sup.b During a Course of Therapy Once every 3 Value
Weekly Weekly weeks No toxicity Maintain dose level .uparw. 25
mg/m.sup.2 up 1 to a maximum dose Maintain dose of 150 mg/m.sup.2
level Neutropenia 1 (1500 to Maintain dose level Maintain dose
level Maintain dose 1999/mm.sup.3) level 2 (1000 to .dwnarw. 25
mg/m.sup.2 Maintain dose level Maintain dose 1499/mm.sup.3) level 3
(550 to Omit dose, then .dwnarw. 25 mg/m.sup.2 when resolved to
.ltoreq. .dwnarw. 25 mg/m.sup.2 .dwnarw. 25 mg/m.sup.2
999/mm.sup.3) grade 2 4 (<500/mm.sup.3 Omit dose, then .dwnarw.
50 mg/m.sup.2 when resolved to .ltoreq. grade 2 .dwnarw. 50
mg/m.sup.2 .dwnarw. 50 mg/m.sup.2 Neutropenic Omit dose, then
.dwnarw. 50 mg/m.sup.2 when resolved .dwnarw. 50 mg/m.sup.2
.dwnarw. 50 mg/m.sup.2 fever (grade 4 neutropenia &.gtoreq.
grade 2 fever) Other Dose modifications for leukopenia,
thrombocytopenia, and also based on NCI toxicity criteria
hematologic and are the same at the start of subsequent courses of
therapy are recommended for neuropenia Toxicities above. Diarrhea 1
(2-3 stools/day > Maintain dose level Maintain dose level
Maintain dose pretx.sup.c) level 2 (4-6 stools/day > .dwnarw. 25
mg/m.sup.2 Maintain dose level Maintain dose pretx.sup.c) level 3
(7-9 stools/day > Omit dose, then .dwnarw. 25 mg/m.sup.2 when
resolved to .ltoreq. .dwnarw. 25 mg/m.sup.2 .dwnarw. 50 mg/m.sup.2
pretx.sup.c) grade 2 4 (=10 stools/day > Omit dose, then
.dwnarw. 50 mg/m.sup.2 when resolved to .ltoreq. .dwnarw. 50
mg/m.sup.2 .dwnarw. 50 mg/m.sup.2 pretx.sup.c) grade 2 Other
nonhemalogic Toxicities 1 Maintain dose level Maintain dose level 2
Maintain dose level Maintain dose level 3 .dwnarw. 25 mg/m.sup.2
.dwnarw. 25 mg/m.sup.2 4 .dwnarw. 50 mg/m.sup.2 .sup.aAll dose
modifications should be based on the worst preceding toxicity
.sup.bNational Cancer Institute Common Toxicity Criteria
.sup.cPretreatment
[0315] Response and Toxicity Assessment: Toxicity will be assessed
by weekly physical examination and blood counts and graded
according to National Cancer Institute Common Toxicity Criteria.
These evaluations and a complete chemistry profile will be repeated
before each treatment.
[0316] Patients will also keep a daily record of their bowel habit
and their use of anti-motility agents. This diary will include the
time of ingestion of PHY906, a recording of the frequency and
consistency of their bowel movements (formed, loose, or watery),
and the anti-motility treatment which was used by the patient to
manage this symptom.
[0317] A research nurse will contact each patient at least once a
week between visits during the first cycle to reinforce
instructions on the management of diarrhea and the completion of
the diary. Overall quality of life including asthenia, nausea,
vomiting, and loss of appetite will also be evaluated using
established FAST methodology.
[0318] A pill count will be made by a pharmacist to each clinical
visit for treatment to assess compliance with PHY906. An evaluation
of disease response will be made after every two treatment cycles.
Response will be defined according to ECOG criteria and will be
assessed in all patients with measurable or evaluable disease but
will not constitute an endpoint in this study.
[0319] Pharmacokinetics of Irinotecan: In selected patients,
pharmacokinetic studies will be performed to assess whether PHY906
affects the metabolism and elimination of irinotecan. In these
patients, the first dose of irinotecan will be given alone (cycle
1/day 1) and the PHY906 will begin on day 2.
[0320] Blood samples will be collected in heparinized tubes
immediately before irinotecan administration, 30, 60, 90 min during
the infusion of irinotecan and 0.5, 1.5, 3.5 and 6 h after the end
of the infusion on cycle 1, day 1, and on cycle 1 day 8. Samples
will be immediately processed with 2.50 .mu.l of plasma added to
500 .mu.l of internal standard solution in polystyrene tubes. The
internal standard solution will consist of camptothecin 50 .mu.g/ml
in acetonitrile acidified with glacial acetic acid, 4.0 ml in 100
ml. The samples will be vortexed for 30 sec, placed into a
40.degree. C. water bath for 15 min, cooled at room temperature and
then mixed with 900 .mu.l of a 25 mM triethylamine buffer (pH 4.2).
The supernatant will be transferred to 1.5 ml Eppendorf tubes,
centrifuged for 4 min at 13,000.times.g in a microcentrifuge, and
an aliquot of the clear supernatant is analyzed by high performance
liquid chromatography (HPLC).
[0321] Chromatographic analysis will be conducted on a Microsorh
C18 (4.5.times.250 mm, 5 .mu.m particle size) reverse phase HPLC
column eluted with 72:28 (v/v) 25 mM TEA/acetonitrile buffer at 1
ml/min utilizing a fluorescence detector with XEX 372 nm and XEm
535 nm (Pharmacia & Upjohn SOP #UPJ-120-5). Maximum plasma
concentration, terminal half-life, and AUC (area under the curve)
will be determined by non-compartmental analysis of the data
utilizing PC-NONLIN software (Scientific Consulting Lexington, Ky.)
and standard pharmacokinetic equations. The Pharmacokinetic studies
will be performed on, cycle 1/day 1 and cycle 1/day 8, to determine
whether prolonged exposure to PHY906 produces a cumulative effect
on the plasma clearance of irinotecan.
Example 19
Effect of PHY906 on Pacreatic Cancer, Hepatocellular Carcinoma, and
Colorectal Cancer
[0322] A. Materials and Methods
[0323] Drugs: Gemcitabine HCl (GEMZAR.RTM.) was purchased from Eli
Lilly and company (Indianapolis, Ind.). .sup.3H-Gemcitabine (14
Ci/mmol, 1 mCi/ml in 50% EtOH) was purchased from Moravek
Biochemicals, Inc. (Brea, Calif.). Tetrahydrouridine (THU) was
purchased from Calbiochem (San Diego, Calif.).
3'-Azido-3'-deoxythymidine (AZT) and 5-fluorouracil (5-FU) were
purchased from Sigma Chemical Co. (St. Louis, Mo.). Capecitabine
(XELODA.RTM., CAP) was purchased from Roche Laboratories Inc.
(Nutley, N.J.). The clinical batch of PHY906 (PHY906-6, FDA 165542)
with 10% excipient was prepared by Sun Ten Pharmaceutical, Inc.
(Taipei, Taiwan). The PHY906 formula is composed of four herbs:
Scutellaria baicalensis Georgi, Paeonia lactiflora Pall., Ziziphus
jujuba Mill and Glycyrrhiza uralensis Fisch., with a relative
weight ratio of 3:2:2:2. The non-clinical batch 906MT with unkown
amounts of excipient was purchased from Min Tong Pharmaceutical
Co., Ltd. (Taichung, Taiwan). Mice: Female BDF-1 and C57BL/6 mice
with body weights between 16 and 20 g (4-6 weeks old) were
purchased from Charles River Laboratories (Wilmington, Mass.). Male
NCr athymic nude mice with body weight between 16 and 20 g (4-6
weeks old) were purchased from Taconic Farms (Garmantown, N.Y.).
The mouse pancreas tumor PAN02 was purchased from the NCI-Frederick
Cancer Research Facility (Frederick, Md.).
[0324] Preparation of Gemcitabine solution: Reconstituted 200 mg of
Gemcitabine HCl powder with 6.66 ml of PBS (phosphate buffered
saline) to final concentration of 30 mg/ml.
[0325] Preparation of .sup.3H-Gemcitabine solution: Ten microliters
(10 .mu.Ci) of .sup.3H-Gemcitabine was dried under a Speedvac to
remove ethanol, then mixed with 250 .mu.l of 30 mg/ml gemcitabine
to get a final solution with radiospecificity of 0.4 mCi/mmol. The
final concentration of this .sup.3H-Gemcitabine solution was 30
mg/ml.
[0326] Preparation of capecitabine solution from capecitabine
tablet: Capecitabine (150 mg/tablet) was dissolved in 40 mM citrate
buffer (pH 6.0) containing 5% gum arabic as the vehicle. The final
solution contains 36 mg/ml of capecitabine.
[0327] Preparation of Herbal Extract From Dry Powder: The
preparation of the herbal extract followed SOP#HERB-001-PHY906.
Briefly, one gram of different batches of PHY906 dry powder,
containing 10% (clinical batch of PHY906) starch excipient, was
added to 10 ml of 80.degree. C. H.sub.2O and incubated at
80.degree. C. for 30 minutes. The supernatant was separated from
the debris by centrifugation (2060 g, 15 min) at room temperature.
The concentration of PHY906 supernatant was calculated as 90 mg/ml
for clinical batch of PHY906 (1 g/10 ml.times.0.9), based on the
dry weight of the dry powder. Since the excipient amount of 906MT
was unknown, the concentration of this preparation was considered
as 100 mg/ml. The herbal extract was diluted to 50 mg/ml with
water, stored at room temperature, and used within 24 hours. Any
residual precipitant that occurred upon standing was vortexed into
a suspension and used to treat the animals.
[0328] Tumor Cells: The human hepatocellular carcinoma HepG2, human
PANC-1 pancreatic cancer and mouse Colon 38 colorectal cancer cell
lines were purchased from the American Type Culture Collection
(Rockville, Md.). The HepG2 and Colon 38 cell lines were routinely
grown in MEME media while PANC-1 cell line was grown in DMEM media,
supplemented with 10% fetal bovine serum (FBS). A small chunk of
mouse PAN02 tumor from NCI was suspended in PBS solution as a total
volume of 0.8 ml. The cells were implanted into the left flank of
mice. Tumor transplantation from mice to mice was performed when
the tumor reached 1500-2000 mm.
[0329] Mouse Tumor Model: Tumor cells (5.times.10.sup.6 cells in
0.1 ml PBS) were transplanted subcutaneously into the left flank of
mice. After 14 days, tumor ranging in size from 300-500 mm.sup.3
was selected for drug studies. The length and width of the each
tumor was measured with sliding calipers. The tumor size was
estimated according to the following formula:
Tumor size(mm.sup.3)=length(mm).times.width(mm).sup.2/2.
[0330] The studies were conducted and the animals were maintained
at the Yale Animal Facility.
[0331] Antitumor Activity of Chemotherapeutic Agents in the
Presence or Absence of PHY906: A total of 20 tumor-bearing mice
were divided into 4 groups (N=5 mice/group):
1) Vehicle
2) PHY906
[0332] 3) Chemotherapeutic agent 4) PHY906+Chemotherapeutic
agent.
[0333] The first day of drug treatment was defined as day 1. PHY906
(500 mg/kg, bid) was administrated orally to the mice 30 min before
chemotherapeutic agents at the days indicated. Chemotherapeutic
agents were given either intraperitoneally or orally at the dose
and schedule indicated. The tumor size, body weight, and mortality
of the mice were monitored daily. Mice were sacrificed when the
tumor size reached 10% of body weight.
[0334] Plasma Handling: The mouse plasma was separated from the
blood by centrifugation at 10,000 g at 4.degree. C. for 10 min.
[0335] a) Capecitabine-containing sample: The plasma (50 .mu.l) was
extracted with 200 .mu.l of acetonitrile in the presence of 10
.mu.l of 5-iodouracil (IU) (50 .mu.g/ml) as an internal standard.
After centrifugation, the supernatant from capecitabine-containing
sample was dried by a speedvac, reconstituted with 50 .mu.l of 0.1
M sodium acetate buffer (pH 4.8), and extracted with 1.3 ml of
water-saturated ethyl acetate. The organic layer was separated
after centrifugation (10,000 g at 4.degree. C. for 10 min), dried
by a speedvac and reconstituted with 100 .mu.l of water.
[0336] b) Gemcitabine-containing plasma: The plasma (50 .mu.l) was
extracted with 200 .mu.l of acetonitrile (ACN) in the presence of 5
.mu.l of 0.1 mM AZT as an internal standard. After centrifugation,
the supernatant was dried by a Speedvac, reconstituted with 100
.mu.l of solution containing 2.5% ACN and 97.5% of 15 mM ammonium
acetate (pH 5.0).
[0337] The resultant analyte was then analyzed by HPLC with UV
absorption and/or radioactivity.
[0338] B. Results
[0339] 1. Pancreatic Cancer
[0340] a) Effect of PHY906 in Antitumor Activity of Different Doses
of Gemcitabine in Murine PAN02 Bearing C57/BL Mice
[0341] To determine the best dosing regimen in the combinational
use of PHY906 and gemcitabine in order to improved anti-tumor
activity of gemcitabine, three different doses of gemcitabine, 150,
300 and 600 mg/kg, in combination with a fixed dose of PHY906 at
500 mg/kg, were studied in C57/BL6 mice bearing PAN02 murine
pancreatic carcinoma (Table 13). As shown in FIGS. 27-29, PHY906
significantly enhanced the antitumor activity of gemcitabine in
PAN02 bearing mice (p<0.001 in all three doses of gemcitabine).
Apparently, mice treated with gemcitabine at 600 mg/kg showed a
better antitumor effect than at the lower gemcitabine doses.
Gemcitabine at dose of 150 mg/kg did not have antitumor activity in
this mouse model so did PHY906 alone.
TABLE-US-00013 TABLE 13 Summary of Treatment Conditions of PHY906
and Gemcitabine in PAN02 Murine Pancreatic Carcinoma Model. Dose of
Starting Dose of PHY906.sup.a Size of Gemcitabine (mg/kg)
Experiment Strain of Tumor (mg/kg) Batch of (po, D1-4 No. Set Mouse
(mm.sup.3) (ip, D1 & 8) PHY906 & 8-11) 1 2 C57/BL6 300-500
150 906-6 500 2 2 C57/BL6 300-500 300 906-6 500 3 2 C57/BL6 300-500
600 906-6 500 4 3 BDF-1 300-500 300 906-6 500 5 3 BDF-1 300-500 600
906-6 500 6 3 BDF-1 300-500 300 906MT 1000 .sup.b .sup.aThe
concentration of PHY906 was calculated based on the exact excipient
amount (10%). .sup.bThe amount of excipient in 906MT was unknown
and the concentration was calculated based on the dry weight of dry
powder.
[0342] b) Effect of PHY906 in Antitumor Activity of Different Doses
of Gemcitabine in Murine PAN02 Bearing BDF-1 Mice
[0343] In addition to the C57/BL6 mice, PHY906 was also tested on
the antitumor activity of gemcitabine in different mouse species.
PAN02 pancreas carcinoma was therefore implanted into BDF-1 mice.
As shown in FIGS. 30-31, an increase of gemcitabine dosage from 300
mg/kg to 600 mg/kg significantly increased the disparity of tumor
growth rates between gemcitabine alone and concomitant use of both
PHY906 and gemcitabine treatments. The results indicated that the
potentiation of PHY906 in antitumor activity of gemcitabine is
obvious, regardless of mouse models. Additionally, a dose of
gemcitabine at 600 mg/kg was selected for the subsequent animal
experiments.
[0344] c) Effect of PHY906 in Antitumor Activity of Gemcitabine in
Human PANC-1 Xenografts
[0345] PHY906 was tested on the antitumor activity of gemcitabine
in human PANC-1 bearing nude mice. As shown in FIG. 32, the
combination of gemcitabine and PHY906 shrank the tumor size
approximately 60% after the first cycle of drug treatment. A dosing
cycle is defined as one week whereas four-day PHY906 treatment plus
three-day rest. The tumor size maintained stable with 60% tumor
shrinkage after the second cycle of combination drug treatment. The
mice experienced up to 20% bodyweight loss upon the starting the
second dosing cycle, as shown in FIG. 33. The third cycle of
combination drug treatment didn't start until day 26 when the tumor
sizes bounced back to the original ones at day 21.
[0346] d) Effect of Non-Clinical Batches of PHY906 on Gemcitabine
in Antitumor Activity in PAN02 Bearing BDF-1 Mice
[0347] Several non-clinical batches of PHY906 from different
manufactures demonstrated different chemical and biological
responses. The questions is whether these in vitro differences
result in different in vivo responses. 906MT, which had very
different chemical and biological activities from the clinical
batch of PHY906, was compared with PHY906 in the antitumor activity
of gemcitabine in PAN02 bearing BDF-1 mice. Since the amount of
excipient in 906MT was unknown, the dose of 906MT was used at 1
g/kg*, based on the dry weight of powder. The dose of gemcitabine
was 300 mg/kg bid on Days 1 and 8. The dosing schedule of 906MT was
the same as for clinical batch PHY906.
[0348] As shown in FIG. 30, PHY906 appeared to have an effect in
the antitumor activity of gemcitabine, but 906MT did not
demonstrate a significant enhancement in the antitumor activity of
gemcitabine, as shown in FIG. 34. These in vivo results are
consistent with other in vitro pharmacological results (data not
shown). The results indicated that any formulation that contains
different ratios or different species of Scutellaria, Glycyrrhiza,
Ziziphus, and Paeonia might not have the same therapeutic effect as
PHY906 in chemotherapy.
[0349] e) Effect of PHY906 on the Pharmacokinetics of Gemcitabine
(dFdC) and Its Metabolite dFdU
[0350] To determine, for a future clinical trial, if PHY906 would
change the metabolism of gemcitabine, a pharmacokinetics analysis
in mice bearing pancreatic cancer was performed. Mice were divided
into three sets of 15: gemcitabine only, gemcitabine plus PHY906,
and vehicle control. Each mouse in the set was bled twice at two
different time points. Plasma was processed as described in
Materials and Methods and analyzed by HPLC with radioactivity
confirmation of peak identity and quantitation. The studies
revealed that there was not a dramatic difference in the levels of
gemcitabine or its metabolite in circulation (dFdU) in the presence
or absence of PHY906, as shown in FIG. 35. Computation of AUCs
(area under concentration curve), C. (maximum drug concentration),
T. (time of maximum drug concentration) and T.sub.1/2 (half life)
of gemcitabine and dFdU in plasma did not indicate a significant
difference of gemcitabine administered alone or in combination with
PHY906, as summarized in Table 14.
TABLE-US-00014 TABLE 14 Effect of PHY906 on the Pharmacokinetic
Parameters of Gemcitabine (dFdC) and Its Metabolite (dFdU) in the
Plasma of PAN02 Bearing BDF-1 Mice (HPLC-UV/VIS Detection Data) AUC
Cmax Tmax T1/2 mg/ml.min) mg/ml) (min) (min) Detection dFdC dFdC +
dFdC dFdC + dFdC dFdC + dFdC dFdC + Compounds Method alone PHY906
alone PHY906 alone PHY906 alone PHY906 dFdC UV 10.8 15.4 0.23 0.25
15 15 27.0 31.0 dFdC Radioactivity 15.6 25.0 0.36 0.40 15 15 26.5
34.2 dFdU UV 3.1 4.4 0.014 0.022 60 120 98.3 69.8 dFdU
Radioactivity 5.8 8.5 0.031 0.048 60 120 93.0 49.2
[0351] f) Effect of PHY906 on Gemcitabine+Oxaliplatin Treated
NCr-Nude Mice Bearing Human PANC-1 Tumor.
[0352] Due to the poor response of monotherapy of gemcitabine in
pancreatic cancer, several combination therapies with gemcitabine
have been conducted in various phases of clinical trials. Among
them is the combination therapy of oxaliplatin and gemcitabine.
[0353] PHY906 was found not only to slow tumor progress with the
combination therapy of oxaliplatin and gemcitabine, but it also
reduced the tumor size to 50% at day 7 of initial drug treatment in
human PANC-1 xenografts mouse model, as shown in FIG. 36.
[0354] 2. Hepatocellular Carcinoma
[0355] a) Effect of PHY906 on the Antitumor Activity of CPT-11 on
Human HepG2 Tumor-Bearing Nude Mice
[0356] Previous studies indicated that PHY906, in combination with
various chemotherapeutic agents, potentiated the antitumor effects
of chemotherapeutic agents in colorectal and pancreatic cancer in
vivo models. Based upon the known pharmacological profiles of herbs
contained in PHY906, it was speculated that the enhancing effects
might act through immunological and/or hematological systems in
normal mice. Therefore, experiments were designed to test the
hypothesis in nude mice, which are deficient in immunological and
hematological systems. Human hepatocellular carcinoma HepG2 cells
were implanted into NCr-nude mice to test the effect of PHY906 on
the antitumor activity of CPT-11. Previous experiments showed that
the maximum tolerable dose of CPT-11 in nude mice was 200 mg/kg,
which was used in this study. CPT-11 (200 mg/kg, i.p.) was given on
day 0. PHY906 was given twice daily at 500 mg/kg starting on day 0
and continued for 8 days. As shown in FIG. 37, PHY906 enhanced the
antitumor effect of CPT-11 on human HepG2 xenografts in nude mice.
However, unlike the observation in BDF-1 mice, PHY906 showed no
beneficial effect on preventing body weight loss or animal death
caused by CPT-11 (data not shown). The fact that PHY906 does not
protect nude mice from weight loss as it does in normal mice
suggests that PHY906 may exert its effects through hematological
and/or immunological systems.
[0357] b) Effect of PHY906 on the Antitumor Activity of
Capecitabine on Human HepG2 Tumor-Bearing Nude Mice
[0358] Capecitabine was originally developed as an orally available
fluoropyrimidine analog that was capable of delivering high
concentrations of the active cytotoxic agent, 5-FU, to tumor
tissues. After oral administration, capecitabine is metabolized to
5-deoxy-5-fluorocytidine (5'-DFCR) by carboxylesterase in the
liver, and then converted to 5'-deoxy-5-fluorouridine (5'-DFUR) by
cytidine deaminase in the liver or tumor. Furthermore, 5'-DFUR is
catalytically activated to 5'-fluorouracil (5-FU) by pyrimidine
phosphorylase, which is rich in the tumor or liver (Berg et al.,
Semin. Oncol., 25: 53-59, 1998; Schwetz et al., JAMA, 286: 2085,
2001). Capecitabine was studied by Lozano et al to treat 55
patients with nonresectable hepatobiliary carcinoma; however, only
a 13% response rate was observed (Aguayo et al., Seminars In
Oncology, 28: 503-513, 2001; Leung et al., Seminars In Oncology,
28: 514-520, 2001; Lozano et al, Oral Capecitabine (XELODA.RTM.)
for the Treatment of Hepatobillary Cancers (Hepatocellular
Carcinoma, Cholangiocarcinoma, and Gallbladder Cancer). 19. 2000.
Proceedings of ASCO.
[0359] Based on the enhancement in antitumor activities of 5-FU and
CPT-11/5-FU/LV by PHY906 in the animal studies, an experiment was
conducted to study whether PHY906 could enhance the antitumor
activity of capecitabine, an oral prodrug of 5-FU. A total of 20
NCr nude mice transplanted with HepG2 human hepatoma cells were
divided into 4 groups (N=5 mice/group): Group (A) vehicle control;
Group (B) treated with PHY906 (500 mg/kg, bid, day 0-3 and 7-10);
Group (C) treated with capecitabine (360 mg/kg, bid.times.14D); and
Group (D) treated with PHY906 (500 mg/kg, bid, days 0-3, 7-10) plus
capecitabine (360 mg/kg, bid.times.14D). The schedule of PHY906
used in this experiment was the same as that in the previous
clinical phase I/IIa trial for the alleviation of CPT-11/5-FU/LV
induced cytotoxicity. PHY906 was found to enhance the antitumor
activity of capecitabine, as shown in FIG. 38. A similar
observation was found with lower doses of capecitabine (data not
shown).
[0360] c) Effect of PHY906 on the Antitumor Activity of Doxorubicin
on Human HepG2 Tumor-Bearing Nude Mice
[0361] To examine whether PHY906 enhances the antitumor activity of
doxorubicin, PHY906 (500 mg/kg, po, bid) was given to in
conjunction with doxorubicin (7.5 mg/kg, qd, D1, 4, 7) to mice with
HepG2 xenografts on days 1-4 and 8-11. As shown in FIG. 39, PHY906
showed enhancement of the antitumor activity of doxorubicin.
[0362] d) Effect of PHY906 on the Antitumor Activity of Thalidomide
on Human HepG2 Tumor-Bearing Nude Mice
[0363] Mice bearing human HepG2 tumor were divided into four
groups: A) control group; B) PHY906 treatment group: mice were
administrated with PHY906 (500 mg/kg) orally twice a day on days
1-4; C) thalidomide group: mice were administrated with thalidomide
(1 g/kg) intraperitoneally once a day for 14 consecutive days; and
D) PHY906 and thalidomide group: PHY906 was given 30 min before
thalidomide. It was found that PHY906 enhanced the antitumor
activity of thalidomide in human HepG2 xenografts mouse model, as
shown in FIG. 40.
[0364] 3. Colorectal Cancer
[0365] U.S. patent application Ser. No. 10/220,876 and PCT
Application PCT/US01/07353 (which are incorporated by reference in
their entirety) indicated that PHY906 enhanced the antitumor
activity of CPT-11, 5-FU, CPT-11/5-FU/LV, L-OddC, and VP-16 in mice
bearing Colon 38 tumor. Other regimens, such as oxaliplatin and
oxaliplatin/5-FU/LV, in colorectal cancer were also investigated.
Oxaliplatin, a synthetic diaminocyclohexane platinum compound and a
third generation drug of cisplatin, causes platinum-DNA adduct
formation and destroys the integrity of the DNA. Oxaliplatin is the
first platinum compound to show efficacy in colorectal cancer
(Raymond et al., Annals of Oncology, 9: 1053-1071, 2004; Mathe et
al., Biomedicine & Pharmacotherapy, 43: 237-250, 2004). The
combination therapy of oxaliplatin/5-FU/LV has recently received
FDA approval as firstline treatment in colorectal cancer patients.
The response rate of oxaliplatin/5-FU/LV (.about.50%) is superior
than that of 5-FU/LV in colorectal cancer patients.
[0366] The effect of PHY906 was examined on the antitumor
activities of oxaliplatin, 5-FU/LV, and oxaliplatin/5-FU/LV in
Colon 38 tumor bearing BDF-1 mice. Mice were divided into eight
groups (N=5 in each group): (A) control vehicle; group (B) treated
with oxaliplatin; group (C) treated with 5-FU/LV; group (D) treated
with oxaliplatin/5-FU/LV; group (E) treated with PHY906; group (F)
treated with PHY906 plus oxaliplatin; group (G) treated with PHY906
plus 5-FU/LV; and group (H) treated with PHY906 plus oxaliplatin
plus 5-FU/LV. The results indicated that PHY906 enhanced the
antitumor activities of oxaliplatin, and oxaliplatin/5-FU/LV, as
shown in FIGS. 41 and 42, respectively. The dose used in the
combination therapy of oxaliplatin/5-FU/LV resulted in 100% of
mortality in animals. However, the data indicated that PHY906 not
only reduced the mortality to 60%, but also delayed the onset of
animal death caused by oxaliplatin/5-FU/LV, as shown in FIG. 43.
None of animals among the drug treated groups, with exception of
that treated with oxaliplatin/5-FU/LV, experienced bodyweight loss
during the treatment. Due to the mortality, it could not be found
out whether PHY906 had a protection against the bodyweight loss
caused by oxaliplatin/5-FU/LV, as shown in FIG. 44.
[0367] C. Discussion
[0368] The effect of PHY906 was examined on the antitumor
activities of oxaliplatin, 5-FU/LV, oxaliplatin/5-FU/LV,
gemcitabine/oxaliplatin, doxorubicin, and thalidomide in mice. The
results indicate that PHY906 enhanced the antitumor activities of
oxaliplatin, oxaliplatin/5-FU/LV, gemcitabine/oxaliplatin,
doxorubicin, and thalidomide as shown in FIGS. 36, 39, 40, 41, and
42. The results also indicate that PHY906 not only reduced the
mortality to 60%, but also delayed the onset of animal death caused
by oxaliplatin/5-FU/LV, as shown in FIG. 43.
[0369] As discussed previously, PHY906 is a traditional Chinese
botanical formulation comprised of four different herbs, and it has
been used for over 1800 years to treat gastrointestinal ailments,
some of which are commonly observed side-effects in cancer patients
undergoing chemotherapy. PHY906 was found to reduce
chemotherapy-induced toxicities, including body weight loss and
mortality, in addition to enhancing the antitumor efficacy of a
broad-spectrum of anticancer agents, such as CPT-11, 5-FU,
CPT-11/5-FU/LV, VP-16, L-OddC, and oxaliplatin in mouse colorectal
cancer models.
[0370] PHY906 was co-administered with either the oral 5-FU prodrug
capecitabine, doxorubicin, thalidomide or CPT-11 in human
hepatocellular xenografts mouse models and with gemcitabine,
oxaliplatin, gemcitabine/oxaliplatin in mouse pancreatic cancer
models. The studies show that PHY906 significantly enhanced the
therapeutic index of the chemotherapeutic agents. The studies also
revealed that co-administration of PHY906 and CPT-11/5-FU/LV and
PHY906 and capecitabine and PHY906 and gemcitabine in animal models
did not alter the pharmacokinetic profile of CPT-11, 5-FU,
capecitabine, gemcitabine, or their respective metabolites.
[0371] The biochemical studies revealed that the PHY906 formulation
possesses a wide range of pharmacological activities. The potential
mechanism(s) of action of PHY906 include (1) enhancement of
cellular uptake of chemotherapeutic agents via inhibition of MDR;
(2) inhibition of NF-.kappa.B activity; (3) inhibition of MMP
activity; and (4) inhibition of angiogenesis.
[0372] These preclinical in vivo studies have provided rationale
for developing PHY906 in the clinical setting. Currently a phase
I/II open-label dose escalation clinical trial has been opened to
patient enrollment to evaluate the role of PHY906 in combination
with capecitabine in the treatment of hepatocellular carcinoma.
[0373] D. Summary
[0374] PHY906, a Chinese medicinal formula consisting of 4
different herbs, has been in use for some 1800 years to treat
gastrointestinal (G1) ailments including diarrhea, nausea and
vomiting. These side effects are quite common in patients
undergoing cancer chemotherapy, thereby raising the possibility of
using PHY906 to alleviate such symptoms in cancer patients and
improve their quality of life. In a tumor-bearing mouse model,
PHY906 significantly decreased host toxicity induced by both
CPT-11-based and oxaliplatin-based chemotherapies by reducing
mortality and loss in body weight. Of note, PHY906 enhanced the
antitumor activity of various chemotherapeutic agents including
CPT-11, CPT-11/5-FU/LV, thalidomide, capecitabine, doxorubicin,
gemcitabine, oxaliplatin, and gemcitabine/oxaliplatin in
colorectal, pancreatic, and liver tumor mouse models. PHY906 did
not significantly change the pharmacokinetics or tissue
distribution of CPT-11-, CPT-11/5-FU/LV-, or capecitabine-based or
gemcitabine treatments. The mechanism of action of PHY906 is
multi-factorial, including modulation of various cytokines,
enhancing cell permeability and transport of active chemical(s),
inhibition of MMP activity, and regulating NF-.kappa.B expression.
A comprehensive panel of chemical and biological fingerprints has
been developed to ensure product consistency of PHY906. This herbal
preparation was studied as a potential cytotoxicity protective
agent in a phase I/IIa clinical trial in colorectal cancer patients
treated with CPT-11/5-FU/LV (Saltz regimen) in firstline or CPT-11
alone in secondline treatment of patients with advanced colorectal
cancer. A second clinical trial is under way to determine whether
PHY906 enhances the antitumor activity of capecitabine in patients
with hepatocellular cancer.
[0375] It should be understood that the foregoing discussion and
examples merely present a detailed description of certain preferred
embodiments. It therefore should be apparent to those of ordinary
skill in the art that various modifications and equivalents can be
made without departing from the spirit and scope of the invention.
All journal articles, other references, patents, and patent
applications that are identified in this patent application are
incorporated by reference in their entirety.
[0376] The foregoing detailed description has been given for
clearness of understanding only and no unnecessary limitations
should be understood therefrom as modifications will be obvious to
those skilled in the art.
[0377] While the invention has been described in connection with
specific embodiments thereof, it will be understood that it is
capable of further modifications and this application is intended
to cover any variations, uses, or adaptations of the invention
following, in general, the principles of the invention and
including such departures from the present disclosure as come
within known or customary practice within the art to which the
invention pertains and as may be applied to the essential features
hereinbefore set forth and as follows in the scope of the appended
claims.
[0378] All applications, patent, and publications referenced herein
are incorporated by reference to the same extent as if each
individual application, patent, and publication was specifically
and individually indicated to be incorporated by reference.
Specifically, the disclosures of WO 01/66123, WO 06/053049, U.S.
Pat. No. 7,025,993, US 2005/0196473, and US 2003/0211180 are
incorporated herein by reference in their entirety for all
purposes. Furthermore, the following references and their contents
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