U.S. patent application number 16/581024 was filed with the patent office on 2020-05-14 for inhibitors of short-chain dehydrogenase activity for modulating hematopoietic stem cells and hematopoiesis.
The applicant listed for this patent is BOARD OF REGENTS OF THE UNIVERSITY TEXAS SYSTEM. Invention is credited to Amar Desai, Sanford Markowitz, Joseph Ready.
Application Number | 20200147063 16/581024 |
Document ID | / |
Family ID | 59387849 |
Filed Date | 2020-05-14 |
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United States Patent
Application |
20200147063 |
Kind Code |
A1 |
Markowitz; Sanford ; et
al. |
May 14, 2020 |
INHIBITORS OF SHORT-CHAIN DEHYDROGENASE ACTIVITY FOR MODULATING
HEMATOPOIETIC STEM CELLS AND HEMATOPOIESIS
Abstract
A method of modulating hematopoietic stem cells and
hematopoiesis includes administering to a subject in need thereof a
15-PGDH inhibitor.
Inventors: |
Markowitz; Sanford; (Pepper
Pike, OH) ; Desai; Amar; (Cleveland, OH) ;
Ready; Joseph; (Carrollton, TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
BOARD OF REGENTS OF THE UNIVERSITY TEXAS SYSTEM |
Dallas |
TX |
US |
|
|
Family ID: |
59387849 |
Appl. No.: |
16/581024 |
Filed: |
September 24, 2019 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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15799307 |
Oct 31, 2017 |
10420752 |
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16581024 |
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15347587 |
Nov 9, 2016 |
9801863 |
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15799307 |
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14395021 |
Oct 16, 2014 |
9790233 |
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PCT/US2013/036790 |
Apr 16, 2013 |
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15347587 |
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15029943 |
Apr 15, 2016 |
9789116 |
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PCT/US2014/060761 |
Oct 15, 2014 |
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15347587 |
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PCT/US2016/021374 |
Mar 8, 2016 |
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15347587 |
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PCT/US2016/027549 |
Apr 14, 2016 |
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15347587 |
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61624670 |
Apr 16, 2012 |
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61891260 |
Oct 15, 2013 |
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61954202 |
Mar 17, 2014 |
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62019597 |
Jul 1, 2014 |
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62043694 |
Aug 29, 2014 |
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62129885 |
Mar 8, 2015 |
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62147305 |
Apr 14, 2015 |
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62252973 |
Nov 9, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00
20130101; A61K 2300/00 20130101; A61K 31/519 20130101; A61K 2300/00
20130101; A61K 31/496 20130101; A61K 31/4365 20130101; A61K 31/444
20130101; A61K 31/496 20130101; A61K 45/06 20130101; A61K 31/519
20130101; A61K 38/193 20130101; A61K 31/495 20130101; A61K 38/193
20130101; A61K 31/444 20130101; A61K 31/5377 20130101; A61K 31/4365
20130101; A61K 31/5377 20130101 |
International
Class: |
A61K 31/4365 20060101
A61K031/4365; A61K 31/495 20060101 A61K031/495; A61K 38/19 20060101
A61K038/19; A61K 31/496 20060101 A61K031/496; A61K 31/519 20060101
A61K031/519; A61K 45/06 20060101 A61K045/06; A61K 31/5377 20060101
A61K031/5377; A61K 31/444 20060101 A61K031/444 |
Goverment Interests
GOVERNMENT FUNDING
[0002] This invention was made with government support under Grant
No. R01CA127306, R01CA127306-03S1, 1P01CA95471-10, AND 5P50CA150964
awarded by The National Institutes of Health. The United States
government has certain rights in the invention.
Claims
1-24. (canceled)
25: A method of increasing neutrophils in a subject in need
thereof, the method comprising administering to the subject a
15-PGDH inhibitor.
26: The method of claim 25, wherein the subject has or is at risk
of at least one of neutropenia, thrombocytopenia, anemia, or
cytopenia.
27: The method of claim 25, wherein the subject has received a
hematopoietic stem cell transplant, bone marrow transplant,
chemotherapy, a myelosuppressive therapy, radiation therapy,
immunosuppressive therapy, and/or viral therapy, and/or has a bone
marrow disease, cancer, viral infection, aplastic anemia,
myelodysplasia, and/or myelofibrosis.
28: The method of claim 25, wherein the subject has undergone a
bone marrow transplant and/or hematopoietic stem cell
transplant.
29: The method of claim 25, wherein the subject has undergone
chemotherapy.
30: The method of claim 25, wherein the subject has aplastic
anemia.
31: The method of claim 25, wherein the 15-PGDH inhibitor is
administered in combination with an immunosuppressive therapy.
32: The method of claim 25, wherein the 15-PGDH inhibitor is
administered in combination with a reduced dose of an
immunosuppressive therapy.
33: The method of claim 25, wherein the 15-PGDH inhibitor has the
following formula (V): ##STR00079## wherein n is 0-2 X.sup.6 is
independently is N or CR.sup.c R.sup.1, R.sup.6, R.sup.7, and
R.sup.c are each independently selected from the group consisting
of hydrogen, substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl containing
from 5-6 ring atoms, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl,
acyloxy, C.sub.2-C.sub.24 alkoxycarbonyl, C.sub.6-C.sub.20
aryloxycarbonyl, C.sub.2-C.sub.24 alkylcarbonato, C.sub.6-C.sub.20
arylcarbonato, carboxy, carboxylato, carbamoyl, C.sub.1-C.sub.24
alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,
isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,
thioformyl, amino, C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20
aryl amino, C.sub.2-C.sub.24 alkylamido, C.sub.6-C.sub.20
arylamido, imino, alkylimino, arylimino, nitro, nitroso, sulfo,
sulfonato, C.sub.1-C.sub.24 alkylsulfanyl, arylsulfanyl,
C.sub.1-C.sub.24 alkylsulfinyl, C.sub.5-C.sub.20 arylsulfinyl,
C.sub.1-C.sub.24 alkylsulfonyl, C.sub.5-C.sub.20 arylsulfonyl,
sulfonamide, phosphono, phosphonato, phosphinato, phospho,
phosphino, polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; and
pharmaceutically acceptable salts thereof.
34: The method of claim 25, wherein the 15-PGDH inhibitor has the
following formula (VI): ##STR00080## wherein n=0-2; X.sup.6 is N or
CR.sup.c; R.sup.1 is selected from the group consisting of branched
or linear alkyl including --(CH.sub.2)n.sub.1CH.sub.3
(n.sub.1=0-7), ##STR00081## wherein n.sub.2=0-6 and X is any of the
following: CF.sub.yH.sub.z (y+z=3), CCl.sub.yH.sub.z (y+z=3), OH,
OAc, OMe, R.sup.71, OR.sup.72, CN, N(R.sup.73).sub.2, ##STR00082##
(n.sub.3=0-5, m=1-5), and ##STR00083## (n.sub.4=0-5). R.sup.5 is
selected from the group consisting of H, Cl, F, NH.sub.2, and
N(R.sup.76).sub.2; R.sup.6 and R.sup.7 can each independently be
one of the following: ##STR00084## ##STR00085## each R.sup.8,
R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14,
R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22, R.sup.23, R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28, R.sup.29, R.sup.30, R.sup.31, R.sup.32,
R.sup.33, R.sup.34, R.sup.35, R.sup.36, R.sup.37, R.sup.38,
R.sup.39, R.sup.40, R.sup.41, R.sup.42, R.sup.43, R.sup.44,
R.sup.45, R.sup.46, R.sup.47, R.sup.48, R.sup.49, R.sup.50,
R.sup.51, R.sup.52, R.sup.53, R.sup.54, R.sup.55, R.sup.56,
R.sup.57, R.sup.58, R.sup.59, R.sup.60, R.sup.61, R.sup.62,
R.sup.63, R.sup.64, R.sup.65, R.sup.66, R.sup.67, R.sup.68,
R.sup.69, R.sup.70, R.sup.71, R.sup.72, R.sup.73, R.sup.74,
R.sup.76, and R.sup.c are the same or different and are
independently selected from the group consisting of hydrogen,
substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl containing
from 5-6 ring atoms, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl,
acyloxy, C.sub.2-C.sub.24 alkoxycarbonyl, C.sub.6-C.sub.20
aryloxycarbonyl, C.sub.2-C.sub.24 alkylcarbonato, C.sub.6-C.sub.20
arylcarbonato, carboxy, carboxylato, carbamoyl, C.sub.1-C.sub.24
alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,
isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,
thioformyl, amino, C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20
aryl amino, C.sub.2-C.sub.24 alkylamido, C.sub.6-C.sub.20
arylamido, imino, alkylimino, arylimino, nitro, nitroso, sulfo,
sulfonato, C.sub.1-C.sub.24 alkylsulfanyl, arylsulfanyl,
C.sub.1-C.sub.24 alkylsulfinyl, C.sub.5-C.sub.20 arylsulfinyl,
C.sub.1-C.sub.24 alkylsulfonyl, C.sub.5-C.sub.20 arylsulfonyl,
sulfonamide, phosphono, phosphonato, phosphinato, phospho,
phosphino, polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, and combinations
thereof, and pharmaceutically acceptable salts thereof.
35: The method of claim 25, wherein the 15-PGDH inhibitor has the
following formula following formula: ##STR00086## or
pharmaceutically acceptable salts thereof.
36: A method of increasing neutrophils in a subject treated with an
immunosuppressive therapy, the method comprising administering to
the subject treated with the immunosuppressive therapy a 15-PGDH
inhibitor.
37: The method of claim 36, wherein the subject has or is at risk
of at least one of neutropenia, thrombocytopenia, anemia, or
cytopenia.
38: The method of claim 36, wherein the subject has received a
hematopoietic stem cell transplant, bone marrow transplant,
chemotherapy, a myelosuppressive therapy, radiation therapy, and/or
viral therapy, and/or has a bone marrow disease, cancer, viral
infection, aplastic anemia, myelodysplasia, and/or
myelofibrosis.
39: The method of claim 36, wherein the subject has undergone a
bone marrow transplant and/or hematopoietic stem cell
transplant.
40: The method of claim 36, wherein the subject has or is at risk
of at least one of neutropenia, thrombocytopenia, anemia, or
cytopenia.
41: The method of claim 36, wherein the 15-PGDH inhibitor has the
following formula (V): ##STR00087## wherein n is 0-2 X.sup.6 is
independently is N or CR.sup.c R.sup.1, R.sup.6, R.sup.7, and
R.sup.c are each independently selected from the group consisting
of hydrogen, substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl containing
from 5-6 ring atoms, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl,
acyloxy, C.sub.2-C.sub.24 alkoxycarbonyl, C.sub.6-C.sub.20
aryloxycarbonyl, C.sub.2-C.sub.24 alkylcarbonato, C.sub.6-C.sub.20
arylcarbonato, carboxy, carboxylato, carbamoyl, C.sub.1-C.sub.24
alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,
isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,
thioformyl, amino, C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20
aryl amino, C.sub.2-C.sub.24 alkylamido, C.sub.6-C.sub.20
arylamido, imino, alkylimino, arylimino, nitro, nitroso, sulfo,
sulfonato, C.sub.1-C.sub.24 alkylsulfanyl, arylsulfanyl,
C.sub.1-C.sub.24 alkylsulfinyl, C.sub.5-C.sub.20 arylsulfinyl,
C.sub.1-C.sub.24 alkylsulfonyl, C.sub.5-C.sub.20 arylsulfonyl,
sulfonamide, phosphono, phosphonato, phosphinato, phospho,
phosphino, polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; and
pharmaceutically acceptable salts thereof.
42: The method of claim 36, wherein the 15-PGDH inhibitor has the
following formula (VI): ##STR00088## wherein n=0-2; X.sup.6 is N or
CR.sup.c; R.sup.1 is selected from the group consisting of branched
or linear alkyl including --(CH.sub.2).sub.n1CH.sub.3
(n.sub.1=0-7), ##STR00089## wherein n.sub.2=0-6 and X is any of the
following: CF.sub.yH.sub.z (y+z=3), CCl.sub.yH.sub.z (y+z=3), OH,
OAc, OMe, R.sup.71, OR.sup.72, CN, N(R.sup.73).sub.2, ##STR00090##
(n.sub.3=0-5, m=1-5), and ##STR00091## (n.sub.4=0-5). R.sup.5 is
selected from the group consisting of H, Cl, F, NH.sub.2, and
N(R.sup.76).sub.2; R.sup.6 and R.sup.7 can each independently be
one of the following: ##STR00092## ##STR00093## each R.sup.8,
R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14,
R.sup.15, R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20,
R.sup.21, R.sup.22, R.sup.23, R.sup.24, R.sup.25, R.sup.26,
R.sup.27, R.sup.28, R.sup.29, R.sup.30, R.sup.31, R.sup.32,
R.sup.33, R.sup.34, R.sup.35, R.sup.36, R.sup.37, R.sup.38,
R.sup.39, R.sup.40, R.sup.41, R.sup.42, R.sup.43, R.sup.44,
R.sup.45, R.sup.46, R.sup.47, R.sup.48, R.sup.49, R.sup.50,
R.sup.51, R.sup.52, R.sup.53, R.sup.54, R.sup.55, R.sup.56,
R.sup.57, R.sup.58, R.sup.59, R.sup.60, R.sup.61, R.sup.62,
R.sup.63, R.sup.64, R.sup.65, R.sup.66, R.sup.67, R.sup.68,
R.sup.69, R.sup.70, R.sup.71, R.sup.72, R.sup.73, R.sup.74,
R.sup.76, and R.sup.c are the same or different and are
independently selected from the group consisting of hydrogen,
substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl containing
from 5-6 ring atoms, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl,
acyloxy, C.sub.2-C.sub.24 alkoxycarbonyl, C.sub.6-C.sub.20
aryloxycarbonyl, C.sub.2-C.sub.24 alkylcarbonato, C.sub.6-C.sub.20
arylcarbonato, carboxy, carboxylato, carbamoyl, C.sub.1-C.sub.24
alkyl-carbamoyl, arylcarbamoyl, thiocarbamoyl, carbamido, cyano,
isocyano, cyanato, isocyanato, isothiocyanato, azido, formyl,
thioformyl, amino, C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20
aryl amino, C.sub.2-C.sub.24 alkylamido, C.sub.6-C.sub.20
arylamido, imino, alkylimino, arylimino, nitro, nitroso, sulfo,
sulfonato, C.sub.1-C.sub.24 alkylsulfanyl, arylsulfanyl,
C.sub.1-C.sub.24 alkylsulfinyl, C.sub.5-C.sub.20 arylsulfinyl,
C.sub.1-C.sub.24 alkylsulfonyl, C.sub.5-C.sub.20 arylsulfonyl,
sulfonamide, phosphono, phosphonato, phosphinato, phospho,
phosphino, polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, and combinations
thereof, and pharmaceutically acceptable salts thereof.
43: The method of claim 36, wherein the 15-PGDH inhibitor has the
following formula following formula: ##STR00094## or
pharmaceutically acceptable salts thereof.
Description
RELATED APPLICATION
[0001] This application is a Continuation of U.S. patent
application Ser. No. 15/347,587, filed Nov. 9, 2016 (now U.S. Pat.
No. 9,801,863), which is a Continuation-in-part of U.S. patent
application Ser. No. 14/395,021, filed Oct. 16, 2014, which is a
National Phase Filing of PCT/US2013/036790, filed Apr. 16, 2013,
which claims priority to U.S. Provisional Application Ser. No.
61/624,670, filed Apr. 16, 2012, this application is also a
Continuation-in-Part of U.S. Ser. No. 15/029,943, filed Apr. 15,
2016, which is a National Phase Filing of PCT/US2014/060761, filed
Oct. 15, 2014, which claims priority to U.S. Provisional
Application Nos. 61/891,260, filed Oct. 15, 2013, 61/954,202, filed
Mar. 17, 2014, 62/019,597, filed Jul. 1, 2014, and 62/043,694,
filed Aug. 29, 2014, this application also claims priority to
PCT/US2016/021374, filed Mar. 8, 2016, which claims priority to
U.S. Provisional Application No. 62/129,885, filed Mar. 8, 2015,
and also claims prirority to PCT/US2016/027549, filed Apr. 14,
2016, which claims priority to U.S. Provisional Application No.
62/147,305, filed Apr. 14, 2015, and this application also claims
priority from U.S. Provisional Application No. 62/252,973, filed
Nov. 9, 2015, the subject matter of which is incorporated herein by
reference in its entirety.
BACKGROUND
[0003] Short-chain dehydrogenases (SCDs) are a family of
dehydrogenases that share only 15% to 30% sequence identity, with
similarity predominantly in the coenzyme binding domain and the
substrate binding domain. In addition to their role in
detoxification of ethanol, SCDs are involved in synthesis and
degradation of fatty acids, steroids, and some prostaglandins, and
are therefore implicated in a variety of disorders, such as lipid
storage disease, myopathy, SCD deficiency, and certain genetic
disorders.
[0004] The SCD, 15-hydroxy-prostaglandin dehydrogenase (15-PGDH),
(hydroxyprostaglandin dehydrogenase 15-(nicotinamide
adeninedinucleotide); 15-PGDH; Enzyme Commission number 1.1.1.141;
encoded by the HPGD gene), represents the key enzyme in the
inactivation of a number of active prostaglandins, leukotrienes and
hydroxyeicosatetraenoic acids (HETEs) (e.g., by catalyzing
oxidation of PGE.sub.2 to 15-keto-prostaglandin E2, 15k-PGE). The
human enzyme is encoded by the HPGD gene and consists of a
homodimer with subunits of a size of 29 kDa. The enzyme belongs to
the evolutionarily conserved superfamily of short-chain
dehydrogenase/reductase enzymes (SDRs), and according to the
recently approved nomenclature for human enzymes, it is named
SDR36C1. Thus far, two forms of 15-PGDH enzyme activity have been
identified, NAD+-dependent type I 15-PGDH that is encoded by the
HPGD gene, and the type II NADP-dependent 15-PGDH, also known as
carbonyl reductase 1 (CBR1, SDR21C1). However, the preference of
CBR1 for NADP and the high Km values of CBR1 for most prostaglandin
suggest that the majority of the in vivo activity can be attributed
to type I 15-PGDH encoded by the HPGD gene, that hereafter, and
throughout all following text, simply denoted as 15-PGDH.
SUMMARY
[0005] Embodiments described herein relate to compositions and
methods for modulating hematopoietic stem cells and hematopoiesis.
As described in the Examples below, it was found that that
inhibitors of short-chain dehydrogenase activity, such as 15-PGDH
inhibitors, can be administered to a subject in need thereof, cells
of the subject, and/or tissue of the subject alone or in
combination with a cytokine to increase and/or mobilize
hematopoietic stem cells and/or neutrophils in the blood, marrow,
and/or tissue of the subject.
[0006] In some embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing neutrophils in the subject. For
example, the hematopoietic cytokine or cell mobilization agent can
include at least one of G-CSF or Plerixafor.
[0007] In other embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing numbers of and/or of mobilizing
peripheral blood hematopoietic stem cells in the subject. For
example, the hematopoietic cytokine or cell mobilization agent can
include at least one of G-CSF or Plerixafor.
[0008] In still other embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing numbers of hematopoietic stem
cells in blood or bone marrow of the subject. For example, the
hematopoietic cytokine or cell mobilization agent can include at
least one of G-CSF or Plerixafor.
[0009] In other embodiments, the 15-PGDH inhibitor can be
administered to a subject and/or tissue of the subject to increase
tissue stem cells. For example, the 15-PGDH inhibitor can be
administered to bone marrow of a subject to increase stem cells in
the subject.
[0010] In still other embodiments, the 15-PGDH inhibitor can be
administered to a tissue graft donor, bone marrow graft donor,
and/or a hematopoietic stem cell donor, and/or a tissue graft,
and/or a bone marrow graft, and/or a hematopoietic stem cell graft,
to increase the fitness of a donor tissue graft, a donor bone
marrow graft, and/or a donor hematopoietic stem cell graft. For
example, the 15-PGDH inhibitor can be administered to a subject,
and/or bone marrow of a subject to increase the fitness of the
marrow as a donor graft, and/or to a preparation of hematopoietic
stem cells of a subject to increase the fitness of the stem cell
preparation as a donor graft, and/or to a preparation of peripheral
blood hematopoietic stem cells of a subject to increase the fitness
of the stem cell preparation as a donor graft, and/or to a
preparation of umbilical cord blood stem cells to increase the
fitness of the stem cell preparation as a donor graft, and/or to a
preparation of umbilical cord blood stem cells to decrease the
number of units of umbilical cord blood required for
transplantation.
[0011] In other embodiments, the 15-PGDH inhibitor can be
administered to a recipient of a tissue graft transplant, bone
marrow transplant, hematopoietic stem cell transplant, and/or of an
umbilical cord stem cell transplant, in order to decrease the
administration of other treatments or growth factors.
[0012] In still other embodiments, the 15-PGDH inhibitor can be
administered to a subject to increase neutrophil counts following a
hematopoietic cell transplant with bone marrow, hematopoietic stem
cells, or umbilical cord blood, to increase neutrophil counts in a
subject with neutropenia following chemotherapy administration or
radiation therapy, to increase neutrophil counts in a subject with
aplastic anemia, myelodysplasia, myelofibrosis, neutropenia due to
other bone marrow diseases, drug induced neutropenia, autoimmune
neutropenia, idiopathic neutropenia, or neutropenia following viral
infections, to increase neutrophil counts in a subject with
neutropenia, to increase platelet counts following a hematopoietic
cell transplant with bone marrow, hematopoietic stem cells, or
umbilical cord blood, to increase platelet counts in a subject with
thrombocytopenia following chemotherapy administration or radiation
therapy, to increase platelet counts in a subject with aplastic
anemia, myelodysplasia, myelofibrosis, thrombocytopenia due to
other bone marrow diseases, drug induced thrombocytopenia,
autoimmune thrombocytopenia, idiopathic thrombocytopenic purpura,
idiopathic thrombocytopenia, or thrombocytopenia following viral
infections, to increase platelet counts in a subject with
thrombocytopenia, to increase red blood cell counts, or hematocrit,
or hemoglobin level, following a hematopoietic cell transplant with
bone marrow, hematopoietic stem cells, or umbilical cord blood, to
increase red blood cell counts, or hematocrit, or hemoglobin level
in a subject with anemia following chemotherapy administration or
radiation therapy, to increase red blood cell counts, or
hematocrit, or hemoglobin level counts in a subject with aplastic
anemia, myelodysplasia, myelofibrosis, anemia due to other disorder
of bone marrow, drug induced anemia, immune mediated anemias,
anemia of chronic disease, anemia following viral infections, or
anemia of unknown cause, to increase red blood cell counts, or
hematocrit, or hemoglobin level in a subject with anemia, to
increase bone marrow stem cells, following a hematopoietic cell
transplant with bone marrow, hematopoietic stem cells, or umbilical
cord blood, to increase bone marrow stem cells in a subject
following chemotherapy administration or radiation therapy, and/or
to increase bone marrow stem cells in a subject with aplastic
anemia, myelodysplasia, myelofibrosis, other disorder of bone
marrow, drug induced cytopenias, immune cytopenias, cytopenias
following viral infections, or cytopenias.
[0013] In other embodiments, the 15-PGDH inhibitor can be
administered to a subject having or at risk of presence of
cytopenia, neutropenia, thrombocytopenia, lymphocytopenia and
anemia to increase responsiveness and/or potentiate cytokines, such
as hematopoietic cytokines. The cytokines can include, for example,
G-CSF, GM-CSF, EPO, IL-3, IL-6, TPO, TPO-RA (thrombopoietin
receptor agonist), and SCF.
[0014] In some embodiments, the subject has received a
hematopoietic stem cell transplant, bone marrow transplant,
chemotherapy, a myelosuppressive therapy, radiation therapy, or
viral therapy and/or has a bone marrow disease, cancer, viral
infection, aplastic anemia, myelodysplasia, and/or
myelofibrosis.
[0015] Other embodiments relate to a method of treating at least
one of neutropenia, thrombocytopenia, anemia, or cytopenia in a
subject in need thereof by administering to the subject a
therapeutically effective amount of a15-PGDH inhibitor. The 15-PGDH
inhibitor can be administered alone or in combination with a
cytokine, hematopoietic cytokine, and/or cell mobilization agent to
the subject, cells of the subject, and/or tissue of the
subject.
[0016] In some embodiments, the 15-PGDH inhibitor can be
administered to a blood, bone marrow, and/or tissue of a subject at
an amount effective to increase prostaglandin levels in the
subject. The 15-PGDH inhibitor can include a compound having
formula (I):
##STR00001## [0017] wherein n is 0-2; [0018] Y.sup.1, Y.sup.2, and
R.sup.1 are the same or different and are each selected from the
group consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O) (C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N), formyl
(--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), phosphono (--P(O)(OH).sub.2), phosphonato
(--P(O)(O.sup.-).sub.2), phosphinato (--P(O)(O.sup.-)), phospho
(--PO.sub.2), phosphino (--PH.sub.2), combinations thereof, and
wherein Y.sup.1 and Y.sup.2 may be linked to form a cyclic or
polycyclic ring, wherein the ring is a substituted or unsubstituted
aryl, a substituted or unsubstituted heteroaryl, a substituted or
unsubstituted cycloalkyl, and a substituted or unsubstituted
heterocyclyl; [0019] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R.sup.1, (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein
R' is H or a lower alkyl group), and wherein R.sup.1 and R.sup.2
may be linked to form a cyclic or polycyclic ring, wherein R.sup.3
and R.sup.4 are same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0020]
X.sup.1 and X.sup.2 are independently N or C, and wherein when
X.sup.1 and/or X.sup.2 are N, Y.sup.1 and/or Y.sup.2, respectively,
are absent; [0021] Z.sup.1 is O, S, CR.sup.aR.sup.b or NR.sup.a,
wherein R.sup.a and R.sup.b are independently H or a C.sub.1-8
alkyl, which is linear, branched, or cyclic, and which is
unsubstituted or substituted; [0022] and pharmaceutically
acceptable salts thereof.
[0023] In other embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (V):
##STR00002## [0024] wherein n is 0-2 [0025] X.sup.6 is
independently is N or CR.sup.c [0026] R.sup.1, R.sup.6, R.sup.7,
and R.sup.c are each independently selected from the group
consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O)(C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH2, --SO.sub.2NY.sub.2
(wherein Y is independently H, aryl or alkyl), phosphono
(--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; [0027] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0028]
and pharmaceutically acceptable salts thereof.
[0029] In some embodiments, R.sup.1 is selected from the group
consisting of branched or linear alkyl including
--(CH.sub.2).sub.n1CH.sub.3 (n.sub.1=0-7),
##STR00003##
wherein n.sub.2=0-6 and X is any of the following: CF.sub.yH.sub.z
(y+z=3), CCl.sub.yH.sub.z (y+z=3), OH, OAc, OMe, R.sup.71,
OR.sup.72, CN, N(R.sup.73).sub.2,
##STR00004##
(n.sub.3=0-5, m=1-5), and
##STR00005##
(n.sub.4=0-5).
[0030] In other embodiments, R.sup.6 and R.sup.7 can each
independently be one of the following:
##STR00006## ##STR00007## [0031] each R.sup.8, R.sup.9, R.sup.10,
R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16,
R.sup.17, R.sup.18, R.sup.19, R.sup.20, R.sup.21, R.sup.22,
R.sup.23, R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28,
R.sup.29, R.sup.30, R.sup.31, R.sup.32, R.sup.33, R.sup.34,
R.sup.35, R.sup.36, R.sup.37, R.sup.38, R.sup.39, R.sup.40,
R.sup.41, R.sup.42, R.sup.43, R.sup.44, R.sup.45, R.sup.46,
R.sup.47, R.sup.48, R.sup.49, R.sup.50, R.sup.51, R.sup.52,
R.sup.53, R.sup.54, R.sup.55, R.sup.56, R.sup.57, R.sup.58,
R.sup.59, R.sup.60, R.sup.61, R.sup.62, R.sup.63, R.sup.64,
R.sup.65, R.sup.66, R.sup.67, R.sup.68, R.sup.69, R.sup.70,
R.sup.71, R.sup.72, R.sup.73, and R.sup.74, are the same or
different and are independently selected from the group consisting
of hydrogen, substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl, C.sub.3-C.sub.2
aryl, heterocycloalkenyl containing from 5-6 ring atoms, (wherein
from 1-3 of the ring atoms is independently selected from N, NH,
N(C.sub.1-C.sub.6 alkyl), NC(O)(C.sub.1-C.sub.6 alkyl), O, and S),
heteroaryl or heterocyclyl containing from 5-14 ring atoms,
(wherein from 1-6 of the ring atoms is independently selected from
N, NH, N(C.sub.1-C.sub.3 alkyl), O, and S), C.sub.6-C.sub.24
alkaryl, C.sub.6-C.sub.24 aralkyl, halo, silyl, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl
(including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), sulfanamido (--SO.sub.2N(R).sub.2
where R is independently H, alkyl, aryl or heteroaryl), imino
(--CR.dbd.NH where R is hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl),
where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkyl ethers (--[(CH.sub.2).sub.nO].sub.m),
phosphates, phosphate esters [--OP(O)(OR).sub.2 where R.dbd.H,
methyl or other alkyl], groups incorporating amino acids or other
moieties expected to bear positive or negative charge at
physiological pH, and combinations thereof, and pharmaceutically
acceptable salts thereof.
[0032] In some embodiments, the 15-PGDH inhibitor can inhibit the
enzymatic activity of recombinant 15-PGDH at an IC.sub.50 of less
than 1 .mu.M, or preferably at an IC.sub.50 of less than 250 nM, or
more preferably at an IC.sub.50 of less than 50 nM, or more
preferably at an IC.sub.50 of less than 10 nM, or more preferably
at an IC.sub.50 of less than 5 nM at a recombinant 15-PGDH
concentration of about 5 nM to about 10 nM.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033] FIG. 1 illustrates a graph showing CFU counts in wild type
bone marrow treated with SW033291 and PGE-2.
[0034] FIGS. 2(A-F) illustrate graphs showing: (A) bone marrow
cellularity of mice treated with SW033291; (B) white blood cell
(WBC) counts in mice treated SW033291; (C) red blood cell (RBC)
counts in mice treated SW033291; (D) platelet counts in mice
treated SW033291; (E) SKL % in whole bone marrow of mice treated
with SW033291; and (F) CFU counts in mice treated SW033291.
[0035] FIGS. 3(A-B) illustrate: (A) a schematic diagram following
CD45.2 antigen marked cells in lethally irradiated C57BL/6J mice
rescued with a bone marrow transplant from donor mice treated with
SW033291 or with vehicle; and (B) graphs showing chimerism, of
donor B-Cells, myeloid cells, and T-Cells after such treatment.
[0036] FIG. 4 illustrates a schematic diagram showing schema of a
study in which C57BL/61 mice are irradiated with 11GY on day 0 and
followed by treatment with SW033291.
[0037] FIGS. 5(A-D) illustrate: (A) a schematic illustration
showing the design of a study of enhanced survival in mice
receiving a bone marrow transplant and also administered the
15-PGDH inhibitor SW033291; (B) graphical survival curves for mice
transplanted with 100,000 donor cells; (C) graphical survival
curves for mice transplanted with 200,000 donor cells; and (D)
graphical survival curves for mice transplanted with 500,000 donor
cells.
[0038] FIGS. 6(A-C) illustrate: (A) a schematic illustration
showing measurements on blood and bone marrow on day 5 after
transplant; (B) a graph showing that SW033291 treated mice have
significantly higher total white count; (C) a graph showing that
SW033291 treated mice have significantly higher total platelet
count. The star symbol denotes P<0.05.
[0039] FIGS. 7(A-B) illustrate: (A) a schematic illustration
showing measurements on blood and bone marrow on day 8 after
transplant; and (B) a graph showing that SW033291 treated mice have
significantly higher platelet count than control, with drug treated
mice having 77,000 platelets compared to control mice having 39,500
platelets. The star symbol denotes P<0.05.
[0040] FIGS. 8(A-D) illustrate: (A) a schematic illustration
showing measurements on blood and bone marrow on day 12 after
transplant; (B) a graph showing that SW033291 treated mice have
significantly higher neutrophil counts, with drug treated mice
having 332 neutrophils compared to control mice having 125
neutrophils; and (C) a graph showing that on day 12 after
transplant, SW033291 treated mice have significantly higher
hemoglobin count than controls, with drug treated mice having
hemoglobin level of 11.58 and control mice having hemoglobin level
of 8.3; and D) a graph showing that SW033291 treated mice have
significantly higher total white counts compared to control mice.
The star symbol denotes P<0.05.
[0041] FIGS. 9(A-G) illustrate: (A) a schematic illustration
showing measurements on blood and bone marrow on day 18 after
transplant; (B-D) graphs showing SW033291 treated mice have
significantly higher total white count (FIG. 85B), lymphocyte count
(FIG. 85C), and neutrophil count (FIG. 85D), with drug treated mice
having 835 neutrophils and control mice having 365 neutrophils
(FIG. 85D); (E) a graph showing that on day 18 drug treated mice
have significantly higher platelet counts than control mice; and
(F-G) graphs showing drug treated mice have nearly 4-fold increased
percentage (FIG. 85F) and total numbers (FIG. 85G) of SKL marked
bone marrow stem cells than do control mice. The star symbol
denotes P<0.05.
[0042] FIGS. 10(A-B) illustrate graphs showing (A) measurement of
PGE2 (pg of PGE2/mg tissue protein) in 4 different mouse tissues
(colon, bone marrow, liver, lung) across time following IP
injection of SW033291 at 10 mg/kg; and (B) time course of PGE2 in
control mice injected with vehicle only.
[0043] FIG. 11 is a schematic illustration showing an experiment in
which mice are lethally irradiated (IR) and 12 hours later receive
a transplant (BMT) with CFSE dye labeled bone marrow cells (BM),
and the number of transplanted cells that home and survive in the
bone marrow of the recipient mice are then determined by FACS at 16
hours post-transplant.
[0044] FIG. 12 illustrates a graph showing the percent of CFSE dye
labeled cells that have homed to the bone marrow of mice treated as
illustrated in FIG. 11.
[0045] FIG. 13 is a schematic illustration showing an experiment in
which mice are lethally irradiated (IR) and 12 hours later receive
a transplant (BMT) with CFSE dye labeled bone marrow cells (BM),
and numbers of transplanted cells that home and survive in the bone
marrow of the recipient mice are then determined by FACS at 16
hours post-transplant.
[0046] FIG. 14 illustrates a graph showing the percent of CFSE dye
labeled cells that have homed to the bone marrow of mice treated as
illustrated in FIG. 13.
[0047] FIGS. 15(A-B) illustrate graphs showing induction of gene
expression in (A) bone marrow SKL cells and (B) bone marrow stromal
cells of mice injected with SW033291 twice daily IP at 10 mg/kg for
3 days.
[0048] FIG. 16 is a schematic illustration showing an experiment in
which immune deficient NSG mice are lethally irradiated (IR) and 12
hours later receive a transplant with CFSE dye labeled buffy coat
cells from human umbilical cord blood (UCB), and number of
transplanted cells that home and survive in the bone marrow of the
recipient mice are then determined by FACS at 16 hours
post-transplant.
[0049] FIG. 17 illustrates a graph showing the percent of CFSE dye
labeled human umbilical cord buffy coat cells that have homed to
the bone marrow of mice treated as per the schema above.
[0050] FIG. 18 illustrates a schematic diagram showing the design
of a study in which 4 groups of mice (4 mice per group) were
administered either A) vehicle control; B) 2.5 mg/kg of 15-PGDH
inhibitor (+) SW209415 twice daily IP for 7 doses; c) recombinant
human G-CSF 250 .mu.g/kg subcutaneously once daily for 4 doses or
d) the combination of 2.5 mg/kg of 15-PGDH inhibitor (+) SW209415
twice daily IP for 7 doses plus recombinant human G-CSF 250
.mu.g/kg subcutaneously once daily for 4 doses (with the daily dose
of G-CSF being administered coincident with a dose of (+)
SW209415).
[0051] FIG. 19 illustrates graphs showing total counts of white
blood cells, neutrophils, and lymphocytes of mice administered
15-PGDH inhibitor (+) SW209415 and/or G-CSF as described in FIG.
18.
[0052] FIG. 20 illustrates graphs showing total counts of red blood
cells, hematocrit, hemoglobin, and platelets of mice administered
15-PGDH inhibitor (+) SW209415 and/or G-CSF as described in FIG.
18.
[0053] FIG. 21 illustrates graphs showing the percent and number of
circulating SKL (Sca-1+; C-kit+; Lin-) marked cells in peripheral
blood of mice administered 15-PGDH inhibitor (+) SW209415 and/or
G-CSF as described in FIG. 18.
[0054] FIG. 22 illustrates graphs showing the percent and number of
SKL (Sca-1+; C-kit+; Lin-) marked cells in bone marrow of mice
administered 15-PGDH inhibitor (+) SW209415 and/or G-CSF as
described in FIG. 18.
[0055] FIGS. 23(A-G) illustrate graphs showing (A) WBC counts, (B)
neutrophil counts, (C) lymphocyte counts, (D) RBC counts, (E)
hemoglobin counts, (F) hematocrit counts, and (G) platelet counts
in peripheral blood of recipient mice lethally irradiated followed
by bone marrow transplantation (BMT) treated with vehicle, G-CSF,
SW209415, and SW209415 in combination with G-CSF at day 8 following
BMT.
[0056] FIGS. 24(A-G) illustrate graphs showing (A) WBC counts, (B)
neutrophil counts, (C) lymphocyte counts, (D) RBC counts, (E)
hemoglobin counts, (F) hematocrit counts, and (G) platelet counts
in peripheral blood of recipient mice lethally irradiated followed
by bone marrow transplantation (BMT) treated with vehicle, G-CSF,
SW209415, and SW209415 in combination with G-CSF at day 12
following BMT.
[0057] FIGS. 25(A-G) illustrate graphs showing (A) WBC counts, (B)
neutrophil counts, (C) lymphocyte counts, (D) RBC counts, (E)
hemoglobin counts, (F) hematocrit counts, and (G) platelet counts
in peripheral blood of recipient mice lethally irradiated followed
by bone marrow transplantation (BMT) treated with vehicle, G-CSF,
SW209415, and SW209415 in combination with G-CSF at day 18
following BMT.
[0058] FIGS. 26(A-C) illustrate graphs showing (A) BM cellularity
18 days post 500 k cell does BMY, (B) SKL levels 18 days post 500 k
cell dose BMT, and (C) SKL/mouse 18 days post 500 k cell dose BMT
treated with vehicle, G-CSF, SW209415, and SW209415 in combination
with G-CSF at day 18 following BMT.
DETAILED DESCRIPTION
[0059] For convenience, certain terms employed in the
specification, examples, and appended claims are collected here.
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 application belongs.
[0060] The articles "a" and "an" are used herein to refer to one or
to more than one (i.e., to at least one) of the grammatical object
of the article. By way of example, "an element" means one element
or more than one element.
[0061] The terms "comprise," "comprising," "include," "including,"
"have," and "having" are used in the inclusive, open sense, meaning
that additional elements may be included. The terms "such as",
"e.g.", as used herein are non-limiting and are for illustrative
purposes only. "Including" and "including but not limited to" are
used interchangeably.
[0062] The term "or" as used herein should be understood to mean
"and/or", unless the context clearly indicates otherwise.
[0063] As used herein, the term "about" or "approximately" refers
to a quantity, level, value, number, frequency, percentage,
dimension, size, amount, weight or length that varies by as much as
15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% to a reference
quantity, level, value, number, frequency, percentage, dimension,
size, amount, weight or length. In one embodiment, the term "about"
or "approximately" refers a range of quantity, level, value,
number, frequency, percentage, dimension, size, amount, weight or
length .+-.15%, .+-.10%, .+-.9%, .+-.8%, .+-.7%, .+-.6%, .+-.5%,
.+-.4%, .+-.3%, .+-.2%, or .+-.1% about a reference quantity,
level, value, number, frequency, percentage, dimension, size,
amount, weight or length.
[0064] It will be noted that the structure of some of the compounds
of the application include asymmetric (chiral) carbon or sulfur
atoms. It is to be understood accordingly that the isomers arising
from such asymmetry are included herein, unless indicated
otherwise. Such isomers can be obtained in substantially pure form
by classical separation techniques and by stereochemically
controlled synthesis. The compounds of this application may exist
in stereoisomeric form, therefore can be produced as individual
stereoisomers or as mixtures.
[0065] The term "isomerism" means compounds that have identical
molecular formulae but that differ in the nature or the sequence of
bonding of their atoms or in the arrangement of their atoms in
space. Isomers that differ in the arrangement of their atoms in
space are termed "stereoisomers". Stereoisomers that are not mirror
images of one another are termed "diastereoisomers", and
stereoisomers that are non-superimposable mirror images are termed
"enantiomers", or sometimes optical isomers. A carbon atom bonded
to four nonidentical substituents is termed a "chiral center"
whereas a sulfur bound to three or four different substitutents,
e.g. sulfoxides or sulfinimides, is likewise termed a "chiral
center".
[0066] The term "chiral isomer" means a compound with at least one
chiral center. It has two enantiomeric forms of opposite chirality
and may exist either as an individual enantiomer or as a mixture of
enantiomers. A mixture containing equal amounts of individual
enantiomeric forms of opposite chirality is termed a "racemic
mixture". A compound that has more than one chiral center has 2n-1
enantiomeric pairs, where n is the number of chiral centers.
Compounds with more than one chiral center may exist as either an
individual diastereomer or as a mixture of diastereomers, termed a
"diastereomeric mixture". When one chiral center is present, a
stereoisomer may be characterized by the absolute configuration (R
or S) of that chiral center. Alternatively, when one or more chiral
centers are present, a stereoisomer may be characterized as (+) or
(-). Absolute configuration refers to the arrangement in space of
the substituents attached to the chiral center. The substituents
attached to the chiral center under consideration are ranked in
accordance with the Sequence Rule of Cahn, Ingold and Prelog. (Cahn
et al, Angew. Chem. Inter. Edit. 1966, 5, 385; errata 511; Cahn et
al., Angew. Chem. 1966, 78, 413; Cahn and Ingold, J Chem. Soc. 1951
(London), 612; Cahn et al., Experientia 1956, 12, 81; Cahn, J.,
Chem. Educ. 1964, 41, 116).
[0067] The term "geometric Isomers" means the diastereomers that
owe their existence to hindered rotation about double bonds. These
configurations are differentiated in their names by the prefixes
cis and trans, or Z and E, which indicate that the groups are on
the same or opposite side of the double bond in the molecule
according to the Cahn-Ingold-Prelog rules. Further, the structures
and other compounds discussed in this application include all
atropic isomers thereof.
[0068] The term "atropic isomers" are a type of stereoisomer in
which the atoms of two isomers are arranged differently in space.
Atropic isomers owe their existence to a restricted rotation caused
by hindrance of rotation of large groups about a central bond. Such
atropic isomers typically exist as a mixture, however as a result
of recent advances in chromatography techniques, it has been
possible to separate mixtures of two atropic isomers in select
cases.
[0069] The terms "crystal polymorphs" or "polymorphs" or "crystal
forms" means crystal structures in which a compound (or salt or
solvate thereof) can crystallize in different crystal packing
arrangements, all of which have the same elemental composition.
Different crystal forms usually have different X-ray diffraction
patterns, infrared spectral, melting points, density hardness,
crystal shape, optical and electrical properties, stability and
solubility. Recrystallization solvent, rate of crystallization,
storage temperature, and other factors may cause one crystal form
to dominate. Crystal polymorphs of the compounds can be prepared by
crystallization under different conditions.
[0070] The term "derivative" refers to compounds that have a common
core structure, and are substituted with various groups as
described herein.
[0071] The term "bioisostere" refers to a compound resulting from
the exchange of an atom or of a group of atoms with another,
broadly similar, atom or group of atoms. The objective of a
bioisosteric replacement is to create a new compound with similar
biological properties to the parent compound. The bioisosteric
replacement may be physicochemically or topologically based.
Examples of carboxylic acid bioisosteres include acyl sulfonimides,
tetrazoles, sulfonates, and phosphonates. See, e.g., Patani and
LaVoie, Chem. Rev. 96, 3147-3176 (1996).
[0072] The phrases "parenteral administration" and "administered
parenterally" are art-recognized terms, and include modes of
administration other than enteral and topical administration, such
as injections, and include, without limitation, intravenous,
intramuscular, intrapleural, intravascular, intrapericardial,
intraarterial, intrathecal, intracapsular, intraorbital,
intracardiac, intradermal, intraperitoneal, transtracheal,
subcutaneous, subcuticular, intra-articular, subcapsular,
subarachnoid, intraspinal and intrastemal injection and
infusion.
[0073] The term "treating" is art-recognized and includes
inhibiting a disease, disorder or condition in a subject, e.g.,
impeding its progress; and relieving the disease, disorder or
condition, e.g., causing regression of the disease, disorder and/or
condition. Treating the disease or condition includes ameliorating
at least one symptom of the particular disease or condition, even
if the underlying pathophysiology is not affected.
[0074] The term "preventing" is art-recognized and includes
stopping a disease, disorder or condition from occurring in a
subject, which may be predisposed to the disease, disorder and/or
condition but has not yet been diagnosed as having it. Preventing a
condition related to a disease includes stopping the condition from
occurring after the disease has been diagnosed but before the
condition has been diagnosed.
[0075] The term "pharmaceutical composition" refers to a
formulation containing the disclosed compounds in a form suitable
for administration to a subject. In a preferred embodiment, the
pharmaceutical composition is in bulk or in unit dosage form. The
unit dosage form is any of a variety of forms, including, for
example, a capsule, an IV bag, a tablet, a single pump on an
aerosol inhaler, or a vial. The quantity of active ingredient
(e.g., a formulation of the disclosed compound or salts thereof) in
a unit dose of composition is an effective amount and is varied
according to the particular treatment involved. One skilled in the
art will appreciate that it is sometimes necessary to make routine
variations to the dosage depending on the age and condition of the
patient. The dosage will also depend on the route of
administration. A variety of routes are contemplated, including
oral, pulmonary, rectal, parenteral, transdermal, subcutaneous,
intravenous, intramuscular, intraperitoneal, intranasal,
inhalational, and the like. Dosage forms for the topical or
transdermal administration of a compound described herein includes
powders, sprays, ointments, pastes, creams, lotions, gels,
solutions, patches, nebulized compounds, and inhalants. In a
preferred embodiment, the active compound is mixed under sterile
conditions with a pharmaceutically acceptable carrier, and with any
preservatives, buffers, or propellants that are required.
[0076] The term "flash dose" refers to compound formulations that
are rapidly dispersing dosage forms.
[0077] The term "immediate release" is defined as a release of
compound from a dosage form in a relatively brief period of time,
generally up to about 60 minutes. The term "modified release" is
defined to include delayed release, extended release, and pulsed
release. The term "pulsed release" is defined as a series of
releases of drug from a dosage form. The term "sustained release"
or "extended release" is defined as continuous release of a
compound from a dosage form over a prolonged period.
[0078] The phrase "pharmaceutically acceptable" is art-recognized.
In certain embodiments, the term includes compositions, polymers
and other materials and/or dosage forms which are, within the scope
of sound medical judgment, suitable for use in contact with the
tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other problem or complication,
commensurate with a reasonable benefit/risk ratio.
[0079] The phrase "pharmaceutically acceptable carrier" is
art-recognized, and includes, for example, pharmaceutically
acceptable materials, compositions or vehicles, such as a liquid or
solid filler, diluent, excipient, solvent or encapsulating
material, involved in carrying or transporting any subject
composition from one organ, or portion of the body, to another
organ, or portion of the body. Each carrier must be "acceptable" in
the sense of being compatible with the other ingredients of a
subject composition and not injurious to the patient. In certain
embodiments, a pharmaceutically acceptable carrier is
non-pyrogenic. Some examples of materials which may serve as
pharmaceutically acceptable carriers include: (1) sugars, such as
lactose, glucose and sucrose; (2) starches, such as corn starch and
potato starch; (3) cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)
powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8)
excipients, such as cocoa butter and suppository waxes; (9) oils,
such as peanut oil, cottonseed oil, sunflower oil, sesame oil,
olive oil, corn oil and soybean oil; (10) glycols, such as
propylene glycol; (11) polyols, such as glycerin, sorbitol,
mannitol and polyethylene glycol; (12) esters, such as ethyl oleate
and ethyl laurate; (13) agar; (14) buffering agents, such as
magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16)
pyrogen-free water; (17) isotonic saline; (18) Ringer's solution;
(19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other
non-toxic compatible substances employed in pharmaceutical
formulations.
[0080] The compounds of the application are capable of further
forming salts. All of these forms are also contemplated herein.
[0081] "Pharmaceutically acceptable salt" of a compound means a
salt that is pharmaceutically acceptable and that possesses the
desired pharmacological activity of the parent compound. For
example, the salt can be an acid addition salt. One embodiment of
an acid addition salt is a hydrochloride salt. The pharmaceutically
acceptable salts can be synthesized from a parent compound that
contains a basic or acidic moiety by conventional chemical methods.
Generally, such salts can be prepared by reacting the free acid or
base forms of these compounds with a stoichiometric amount of the
appropriate base or acid in water or in an organic solvent, or in a
mixture of the two; generally, non-aqueous media like ether, ethyl
acetate, ethanol, isopropanol, or acetonitrile being preferred.
Lists of salts are found in Remington's Pharmaceutical Sciences,
18th ed. (Mack Publishing Company, 1990).
[0082] The compounds described herein can also be prepared as
esters, for example pharmaceutically acceptable esters. For
example, a carboxylic acid function group in a compound can be
converted to its corresponding ester, e.g., a methyl, ethyl, or
other ester. Also, an alcohol group in a compound can be converted
to its corresponding ester, e.g., an acetate, propionate, or other
ester.
[0083] The compounds described herein can also be prepared as
prodrugs, for example pharmaceutically acceptable prodrugs. The
terms "pro-drug" and "prodrug" are used interchangeably herein and
refer to any compound, which releases an active parent drug in
vivo. Since prodrugs are known to enhance numerous desirable
qualities of pharmaceuticals (e.g., solubility, bioavailability,
manufacturing, etc.) the compounds can be delivered in prodrug
form. Thus, the compounds described herein are intended to cover
prodrugs of the presently claimed compounds, methods of delivering
the same and compositions containing the same. "Prodrugs" are
intended to include any covalently bonded carriers that release an
active parent drug in vivo when such prodrug is administered to a
subject. Prodrugs are prepared by modifying functional groups
present in the compound in such a way that the modifications are
cleaved, either in routine manipulation or in vivo, to the parent
compound. Prodrugs include compounds wherein a hydroxy, amino,
sulfhydryl, carboxy, or carbonyl group is bonded to any group that
may be cleaved in vivo to form a free hydroxyl, free amino, free
sulfhydryl, free carboxy or free carbonyl group, respectively.
Prodrugs can also include a precursor (forerunner) of a compound
described herein that undergoes chemical conversion by metabolic
processes before becoming an active or more active pharmacological
agent or active compound described herein.
[0084] Examples of prodrugs include, but are not limited to, esters
(e.g., acetate, dialkylaminoacetates, formates, phosphates,
sulfates, and benzoate derivatives) and carbamates (e.g.,
N,N-dimethylaminocarbonyl) of hydroxy functional groups, ester
groups (e.g., ethyl esters, morpholinoethanol esters) of carboxyl
functional groups, N-acyl derivatives (e.g., N-acetyl)N-Mannich
bases, Schiff bases and enaminones of amino functional groups,
oximes, acetals, ketals and enol esters of ketone and aldehyde
functional groups in compounds, and the like, as well as sulfides
that are oxidized to form sulfoxides or sulfones.
[0085] The term "protecting group" refers to a grouping of atoms
that when attached to a reactive group in a molecule masks, reduces
or prevents that reactivity. Examples of protecting groups can be
found in Green and Wuts, Protective Groups in Organic Chemistry,
(Wiley, 2.sup.nd ed. 1991); Harrison and Harrison et al.,
Compendium of Synthetic Organic Methods, Vols. 1-8 (John Wiley and
Sons, 1971-1996); and Kocienski, Protecting Groups, (Verlag,
3.sup.rd ed. 2003).
[0086] The term "amine protecting group" is intended to mean a
functional group that converts an amine, amide, or other
nitrogen-containing moiety into a different chemical group that is
substantially inert to the conditions of a particular chemical
reaction. Amine protecting groups are preferably removed easily and
selectively in good yield under conditions that do not affect other
functional groups of the molecule. Examples of amine protecting
groups include, but are not limited to, formyl, acetyl, benzyl,
t-butyldimethylsilyl, t-butyldiphenylsilyl, t-butyloxycarbonyl
(Boc), p-methoxybenzyl, methoxymethyl, tosyl, trifluoroacetyl,
trimethylsilyl (TMS), fluorenyl-methyloxycarbonyl,
2-trimethylsilyl-ethyoxycarbonyl, 1-methyl-1-(4-biphenylyl)
ethoxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl (CBZ),
2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted
trityl groups, 9-fluorenylmethyloxycarbonyl (FMOC),
nitro-veratryloxycarbonyl (NVOC), and the like. Those of skill in
the art can identify other suitable amine protecting groups.
[0087] Representative hydroxy protecting groups include those where
the hydroxy group is either acylated or alkylated such as benzyl,
and trityl ethers as well as alkyl ethers, tetrahydropyranyl
ethers, trialkylsilyl ethers and allyl ethers.
[0088] Additionally, the salts of the compounds described herein,
can exist in either hydrated or unhydrated (the anhydrous) form or
as solvates with other solvent molecules. Non-limiting examples of
hydrates include monohydrates, dihydrates, etc. Nonlimiting
examples of solvates include ethanol solvates, acetone solvates,
etc.
[0089] The term "solvates" means solvent addition forms that
contain either stoichiometric or non-stoichiometric amounts of
solvent. Some compounds have a tendency to trap a fixed molar ratio
of solvent molecules in the crystalline solid state, thus forming a
solvate. If the solvent is water the solvate formed is a hydrate,
when the solvent is alcohol, the solvate formed is an alcoholate.
Hydrates are formed by the combination of one or more molecules of
water with one of the substances in which the water retains its
molecular state as H.sub.2O, such combination being able to form
one or more hydrate.
[0090] The compounds, salts and prodrugs described herein can exist
in several tautomeric forms, including the enol and imine form, and
the keto and enamine form and geometric isomers and mixtures
thereof. Tautomers exist as mixtures of a tautomeric set in
solution. In solid form, usually one tautomer predominates. Even
though one tautomer may be described, the present application
includes all tautomers of the present compounds. A tautomer is one
of two or more structural isomers that exist in equilibrium and are
readily converted from one isomeric form to another. This reaction
results in the formal migration of a hydrogen atom accompanied by a
switch of adjacent conjugated double bonds. In solutions where
tautomerization is possible, a chemical equilibrium of the
tautomers will be reached. The exact ratio of the tautomers depends
on several factors, including temperature, solvent, and pH. The
concept of tautomers that are interconvertable by tautomerizations
is called tautomerism.
[0091] Of the various types of tautomerism that are possible, two
are commonly observed. In keto-enol tautomerism a simultaneous
shift of electrons and a hydrogen atom occurs.
[0092] Tautomerizations can be catalyzed by: Base: 1.
deprotonation; 2. formation of a delocalized anion (e.g., an
enolate); 3. protonation at a different position of the anion;
Acid: 1. protonation; 2. formation of a delocalized cation; 3.
deprotonation at a different position adjacent to the cation.
[0093] The term "analogue" refers to a chemical compound that is
structurally similar to another but differs slightly in composition
(as in the replacement of one atom by an atom of a different
element or in the presence of a particular functional group, or the
replacement of one functional group by another functional group).
Thus, an analogue is a compound that is similar or comparable in
function and appearance, but not in structure or origin to the
reference compound.
[0094] A "patient," "subject," or "host" to be treated by the
subject method may mean either a human or non-human animal, such as
a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the
subject of the herein disclosed methods can be a human, non-human
primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig
or rodent. The term does not denote a particular age or sex. Thus,
adult and newborn subjects, as well as fetuses, whether male or
female, are intended to be covered. In one aspect, the subject is a
mammal. A patient refers to a subject afflicted with a disease or
disorder.
[0095] The terms "prophylactic" or "therapeutic" treatment is
art-recognized and includes administration to the host of one or
more of the subject compositions. If it is administered prior to
clinical manifestation of the unwanted condition (e.g., disease or
other unwanted state of the host animal) then the treatment is
prophylactic, i.e., it protects the host against developing the
unwanted condition, whereas if it is administered after
manifestation of the unwanted condition, the treatment is
therapeutic (i.e., it is intended to diminish, ameliorate, or
stabilize the existing unwanted condition or side effects
thereof).
[0096] The terms "therapeutic agent", "drug", "medicament" and
"bioactive substance" are art-recognized and include molecules and
other agents that are biologically, physiologically, or
pharmacologically active substances that act locally or
systemically in a patient or subject to treat a disease or
condition. The terms include without limitation pharmaceutically
acceptable salts thereof and prodrugs. Such agents may be acidic,
basic, or salts; they may be neutral molecules, polar molecules, or
molecular complexes capable of hydrogen bonding; they may be
prodrugs in the form of ethers, esters, amides and the like that
are biologically activated when administered into a patient or
subject.
[0097] The phrase "therapeutically effective amount" or
"pharmaceutically effective amount" is an art-recognized term. In
certain embodiments, the term refers to an amount of a therapeutic
agent that produces some desired effect at a reasonable
benefit/risk ratio applicable to any medical treatment. In certain
embodiments, the term refers to that amount necessary or sufficient
to eliminate, reduce or maintain a target of a particular
therapeutic regimen. The effective amount may vary depending on
such factors as the disease or condition being treated, the
particular targeted constructs being administered, the size of the
subject or the severity of the disease or condition. One of
ordinary skill in the art may empirically determine the effective
amount of a particular compound without necessitating undue
experimentation. In certain embodiments, a therapeutically
effective amount of a therapeutic agent for in vivo use will likely
depend on a number of factors, including: the rate of release of an
agent from a polymer matrix, which will depend in part on the
chemical and physical characteristics of the polymer; the identity
of the agent; the mode and method of administration; and any other
materials incorporated in the polymer matrix in addition to the
agent.
[0098] The term "ED50" is art-recognized. In certain embodiments,
ED50 means the dose of a drug, which produces 50% of its maximum
response or effect, or alternatively, the dose, which produces a
pre-determined response in 50% of test subjects or preparations.
The term "LD50" is art-recognized. In certain embodiments, LD50
means the dose of a drug, which is lethal in 50% of test subjects.
The term "therapeutic index" is an art-recognized term, which
refers to the therapeutic index of a drug, defined as
LD50/ED50.
[0099] The terms "IC.sub.50," or "half maximal inhibitory
concentration" is intended to refer to the concentration of a
substance (e.g., a compound or a drug) that is required for 50%
inhibition of a biological process, or component of a process,
including a protein, subunit, organelle, ribonucleoprotein,
etc.
[0100] With respect to any chemical compounds, the present
application is intended to include all isotopes of atoms occurring
in the present compounds. Isotopes include those atoms having the
same atomic number but different mass numbers. By way of general
example and without limitation, isotopes of hydrogen include
tritium and deuterium, and isotopes of carbon include C-13 and
C-14.
[0101] When a bond to a substituent is shown to cross a bond
connecting two atoms in a ring, then such substituent can be bonded
to any atom in the ring. When a substituent is listed without
indicating the atom via which such substituent is bonded to the
rest of the compound of a given formula, then such substituent can
be bonded via any atom in such substituent. Combinations of
substituents and/or variables are permissible, but only if such
combinations result in stable compounds.
[0102] When an atom or a chemical moiety is followed by a
subscripted numeric range (e.g., C.sub.1-6), it is meant to
encompass each number within the range as well as all intermediate
ranges. For example, "C.sub.1-6 alkyl" is meant to include alkyl
groups with 1, 2, 3, 4, 5, 6, 1-6, 1-5, 1-4, 1-3, 1-2, 2-6, 2-5,
2-4, 2-3, 3-6, 3-5, 3-4, 4-6, 4-5, and 5-6 carbons.
[0103] The term "alkyl" is intended to include both branched (e.g.,
isopropyl, tert-butyl, isobutyl), straight-chain e.g., methyl,
ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl),
and cycloalkyl (e.g., alicyclic) groups (e.g., cyclopropyl,
cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), alkyl
substituted cycloalkyl groups, and cycloalkyl substituted alkyl
groups. Such aliphatic hydrocarbon groups have a specified number
of carbon atoms. For example, C.sub.1-6 alkyl is intended to
include C.sub.1, C.sub.2, C.sub.3, C.sub.4, C.sub.5, and C.sub.6
alkyl groups. As used herein, "lower alkyl" refers to alkyl groups
having from 1 to 6 carbon atoms in the backbone of the carbon
chain. "Alkyl" further includes alkyl groups that have oxygen,
nitrogen, sulfur or phosphorous atoms replacing one or more
hydrocarbon backbone carbon atoms. In certain embodiments, a
straight chain or branched chain alkyl has six or fewer carbon
atoms in its backbone (e.g., C.sub.1-C.sub.6 for straight chain,
C.sub.3-C.sub.6 for branched chain), for example four or fewer.
Likewise, certain cycloalkyls have from three to eight carbon atoms
in their ring structure, such as five or six carbons in the ring
structure.
[0104] The term "substituted alkyls" refers to alkyl moieties
having substituents replacing a hydrogen on one or more carbons of
the hydrocarbon backbone. Such substituents can include, for
example, alkyl, alkenyl, alkynyl, halogen, hydroxyl,
alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,
alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl,
dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate,
phosphonato, phosphinato, cyano, amino (including alkylamino,
dialkylamino, arylamino, diarylamino, and alkylarylamino),
acylamino (including alkylcarbonylamino, arylcarbonylamino,
carbamoyl and ureido), amidino, imino, sulfhydryl, alkylthio,
arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,
sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,
heterocyclyl, alkylaryl, or an aromatic or heteroaromatic moiety.
Cycloalkyls can be further substituted, e.g., with the substituents
described above. An "alkylaryl" or an "aralkyl" moiety is an alkyl
substituted with an aryl (e.g., phenylmethyl (benzyl)). If not
otherwise indicated, the terms "alkyl" and "lower alkyl" include
linear, branched, cyclic, unsubstituted, substituted, and/or
heteroatom-containing alkyl or lower alkyl, respectively.
[0105] The term "alkenyl" refers to a linear, branched or cyclic
hydrocarbon group of 2 to about 24 carbon atoms containing at least
one double bond, such as ethenyl, n-propenyl, isopropenyl,
n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl,
eicosenyl, tetracosenyl, cyclopentenyl, cyclohexenyl, cyclooctenyl,
and the like. Generally, although again not necessarily, alkenyl
groups can contain 2 to about 18 carbon atoms, and more
particularly 2 to 12 carbon atoms. The term "lower alkenyl" refers
to an alkenyl group of 2 to 6 carbon atoms, and the specific term
"cycloalkenyl" intends a cyclic alkenyl group, preferably having 5
to 8 carbon atoms. The term "substituted alkenyl" refers to alkenyl
substituted with one or more substituent groups, and the terms
"heteroatom-containing alkenyl" and "heteroalkenyl" refer to
alkenyl or heterocycloalkenyl (e.g., heterocylcohexenyl) in which
at least one carbon atom is replaced with a heteroatom. If not
otherwise indicated, the terms "alkenyl" and "lower alkenyl"
include linear, branched, cyclic, unsubstituted, substituted,
and/or heteroatom-containing alkenyl and lower alkenyl,
respectively.
[0106] The term "alkynyl" refers to a linear or branched
hydrocarbon group of 2 to 24 carbon atoms containing at least one
triple bond, such as ethynyl, n-propynyl, and the like. Generally,
although again not necessarily, alkynyl groups can contain 2 to
about 18 carbon atoms, and more particularly can contain 2 to 12
carbon atoms. The term "lower alkynyl" intends an alkynyl group of
2 to 6 carbon atoms. The term "substituted alkynyl" refers to
alkynyl substituted with one or more substituent groups, and the
terms "heteroatom-containing alkynyl" and "heteroalkynyl" refer to
alkynyl in which at least one carbon atom is replaced with a
heteroatom. If not otherwise indicated, the terms "alkynyl" and
"lower alkynyl" include linear, branched, unsubstituted,
substituted, and/or heteroatom-containing alkynyl and lower
alkynyl, respectively.
[0107] The terms "alkyl", "alkenyl", and "alkynyl" are intended to
include moieties which are diradicals, i.e., having two points of
attachment. A nonlimiting example of such an alkyl moiety that is a
diradical is --CH.sub.2CH.sub.2--, i.e., a C.sub.2 alkyl group that
is covalently bonded via each terminal carbon atom to the remainder
of the molecule.
[0108] The term "alkoxy" refers to an alkyl group bound through a
single, terminal ether linkage; that is, an "alkoxy" group may be
represented as --O-alkyl where alkyl is as defined above. A "lower
alkoxy" group intends an alkoxy group containing 1 to 6 carbon
atoms, and includes, for example, methoxy, ethoxy, n-propoxy,
isopropoxy, t-butyloxy, etc. Preferred substituents identified as
"C.sub.1-C.sub.6 alkoxy" or "lower alkoxy" herein contain 1 to 3
carbon atoms, and particularly preferred such substituents contain
1 or 2 carbon atoms (i.e., methoxy and ethoxy).
[0109] The term "aryl" refers to an aromatic substituent containing
a single aromatic ring or multiple aromatic rings that are fused
together, directly linked, or indirectly linked (such that the
different aromatic rings are bound to a common group such as a
methylene or ethylene moiety). Aryl groups can contain 5 to 20
carbon atoms, and particularly preferred aryl groups can contain 5
to 14 carbon atoms. Examples of aryl groups include benzene,
phenyl, pyrrole, furan, thiophene, thiazole, isothiazole,
imidazole, triazole, tetrazole, pyrazole, oxazole, isooxazole,
pyridine, pyrazine, pyridazine, and pyrimidine, and the like.
Furthermore, the term "aryl" includes multicyclic aryl groups,
e.g., tricyclic, bicyclic, e.g., naphthalene, benzoxazole,
benzodioxazole, benzothiazole, benzoimidazole, benzothiophene,
methylenedioxyphenyl, quinoline, isoquinoline, napthridine, indole,
benzofuran, purine, benzofuran, deazapurine, or indolizine. Those
aryl groups having heteroatoms in the ring structure may also be
referred to as "aryl heterocycles", "heterocycles," "heteroaryls"
or "heteroaromatics". The aromatic ring can be substituted at one
or more ring positions with such substituents as described above,
as for example, halogen, hydroxyl, alkoxy, alkylcarbonyloxy,
arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy,
carboxylate, alkylcarbonyl, alkylaminocarbonyl,
aralkylaminocarbonyl, alkenylaminocarbonyl, alkylcarbonyl,
arylcarbonyl, aralkylcarbonyl, alkenylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkylamino, dialkylamino,
arylamino, diaryl amino, and al kylaryl amino), acylamino
(including alkylcarbonylamino, arylcarbonylamino, carbamoyl and
ureido), amidino, imino, sulfhydryl, alkylthio, arylthio,
thiocarboxylate, sulfates, alkylsulfinyl, sulfonato, sulfamoyl,
sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocyclyl,
alkylaryl, or an aromatic or heteroaromatic moiety. Aryl groups can
also be fused or bridged with alicyclic or heterocyclic rings,
which are not aromatic so as to form a multicyclic system (e.g.,
tetralin, methylenedioxyphenyl). If not otherwise indicated, the
term "aryl" includes unsubstituted, substituted, and/or
heteroatom-containing aromatic substituents.
[0110] The term "alkaryl" refers to an aryl group with an alkyl
substituent, and the term "aralkyl" refers to an alkyl group with
an aryl substituent, wherein "aryl" and "alkyl" are as defined
above. Exemplary aralkyl groups contain 6 to 24 carbon atoms, and
particularly preferred aralkyl groups contain 6 to 16 carbon atoms.
Examples of aralkyl groups include, without limitation, benzyl,
2-phenyl-ethyl, 3-phenyl-propyl, 4-phenyl-butyl, 5-phenyl-pentyl,
4-phenylcyclohexyl, 4-benzylcyclohexyl, 4-phenylcyclohexylmethyl,
4-benzylcyclohexylmethyl, and the like. Alkaryl groups include, for
example, p-methylphenyl, 2,4-dimethylphenyl, p-cyclohexylphenyl,
2,7-dimethylnaphthyl, 7-cyclooctylnaphthyl,
3-ethyl-cyclopenta-1,4-diene, and the like.
[0111] The terms "heterocyclyl" or "heterocyclic group" include
closed ring structures, e.g., 3- to 10-, or 4- to 7-membered rings,
which include one or more heteroatoms. "Heteroatom" includes atoms
of any element other than carbon or hydrogen. Examples of
heteroatoms include nitrogen, oxygen, sulfur and phosphorus.
[0112] Heterocyclyl groups can be saturated or unsaturated and
include pyrrolidine, oxolane, thiolane, piperidine, piperazine,
morpholine, lactones, lactams, such as azetidinones and
pyrrolidinones, sultams, and sultones. Heterocyclic groups such as
pyrrole and furan can have aromatic character. They include fused
ring structures, such as quinoline and isoquinoline. Other examples
of heterocyclic groups include pyridine and purine. The
heterocyclic ring can be substituted at one or more positions with
such substituents as described above, as for example, halogen,
hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy,
aryloxycarbonyloxy, carboxylate, alkylcarbonyl, alkoxycarbonyl,
aminocarbonyl, alkylthiocarbonyl, alkoxyl, phosphate, phosphonato,
phosphinato, cyano, amino (including alkyl amino, dialkylamino,
arylamino, diarylamino, and alkylarylamino), acylamino (including
alkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido),
amidino, imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,
sulfates, sulfonato, sulfamoyl, sulfonamido, nitro,
trifluoromethyl, cyano, azido, heterocyclyl, or an aromatic or
heteroaromatic moiety. Heterocyclic groups can also be substituted
at one or more constituent atoms with, for example, a lower alkyl,
a lower alkenyl, a lower alkoxy, a lower alkylthio, a lower
alkylamino, a lower alkylcarboxyl, a nitro, a hydroxyl, --CF.sub.3,
or --CN, or the like.
[0113] The term "halo" or "halogen" refers to fluoro, chloro,
bromo, and iodo. "Counterion" is used to represent a small,
negatively charged species such as fluoride, chloride, bromide,
iodide, hydroxide, acetate, and sulfate. The term sulfoxide refers
to a sulfur attached to 2 different carbon atoms and one oxygen and
the S--O bond can be graphically represented with a double bond
(S.dbd.O), a single bond without charges (S--O) or a single bond
with charges [S(+)--O(-)].
[0114] The terms "substituted" as in "substituted alkyl,"
"substituted aryl," and the like, as alluded to in some of the
aforementioned definitions, is meant that in the alkyl, aryl, or
other moiety, at least one hydrogen atom bound to a carbon (or
other) atom is replaced with one or more non-hydrogen substituents.
Examples of such substituents include, without limitation:
functional groups such as halo, hydroxyl, silyl, sulfhydryl,
C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl
(including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), mono-(C.sub.1-C.sub.24
alkyl)-substituted carbamoyl (--(CO)--NH(C.sub.1-C.sub.24 alkyl)),
di-(C.sub.1-C.sub.4 alkyl)-substituted carbamoyl
(--(CO)--N(C.sub.1-C.sub.24 alkyl).sub.2), mono-substituted
arylcarbamoyl (--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2),
carbamido (--NH--(CO)--NH.sub.2), cyano(-CN), isocyano
(--N.sup.+C.sup.-), cyanato (--O--CN), isocyanato
(--ON.sup.+C.sup.-), isothiocyanato (--S--CN), azido
(--N.dbd.N.sup.+.dbd.N.sup.-), formyl (--(CO)--H), thioformyl
(--(CS)--H), amino (--NH.sub.2), mono- and di-(C.sub.1-C.sub.24
alkyl)-substituted amino, mono- and di-(C.sub.5-C.sub.20
aryl)-substituted amino, C.sub.2-C.sub.24 alkylamido
(--NH--(CO)-alkyl), C.sub.6-C.sub.20 arylamido (--NH--(CO)-aryl),
imino (--CR.dbd.NH where R=hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), phosphono (--P(O)(OH).sub.2), phosphonato
(--P(O)(O.sup.-).sub.2), phosphinato (--P(O)(O.sup.-)), phospho
(--PO.sub.2), and phosphino (--PH.sub.2); and the hydrocarbyl
moieties C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl,
C.sub.2-C.sub.24 alkynyl, C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24
alkaryl, and C.sub.6-C.sub.24 aralkyl.
[0115] In addition, the aforementioned functional groups may, if a
particular group permits, be further substituted with one or more
additional functional groups or with one or more hydrocarbyl
moieties such as those specifically enumerated above. Analogously,
the above-mentioned hydrocarbyl moieties may be further substituted
with one or more functional groups or additional hydrocarbyl
moieties such as those specifically enumerated.
[0116] When the term "substituted" appears prior to a list of
possible substituted groups, it is intended that the term apply to
every member of that group. For example, the phrase "substituted
alkyl, alkenyl, and aryl" is to be interpreted as "substituted
alkyl, substituted alkenyl, and substituted aryl." Analogously,
when the term "heteroatom-containing" appears prior to a list of
possible heteroatom-containing groups, it is intended that the term
apply to every member of that group. For example, the phrase
"heteroatom-containing alkyl, alkenyl, and aryl" is to be
interpreted as "heteroatom-containing alkyl, substituted alkenyl,
and substituted aryl.
[0117] "Optional" or "optionally" means that the subsequently
described circumstance may or may not occur, so that the
description includes instances where the circumstance occurs and
instances where it does not. For example, the phrase "optionally
substituted" means that a non-hydrogen substituent may or may not
be present on a given atom, and, thus, the description includes
structures wherein a non-hydrogen substituent is present and
structures wherein a non-hydrogen substituent is not present.
[0118] The terms "stable compound" and "stable structure" are meant
to indicate a compound that is sufficiently robust to survive
isolation, and as appropriate, purification from a reaction
mixture, and formulation into an efficacious therapeutic agent.
[0119] The terms "free compound" is used herein to describe a
compound in the unbound state.
[0120] Throughout the description, where compositions are described
as having, including, or comprising, specific components, it is
contemplated that compositions also consist essentially of, or
consist of, the recited components. Similarly, where methods or
processes are described as having, including, or comprising
specific process steps, the processes also consist essentially of,
or consist of, the recited processing steps. Further, it should be
understood that the order of steps or order for performing certain
actions is immaterial so long as the compositions and methods
described herein remains operable. Moreover, two or more steps or
actions can be conducted simultaneously.
[0121] The term "small molecule" is an art-recognized term. In
certain embodiments, this term refers to a molecule, which has a
molecular weight of less than about 2000 amu, or less than about
1000 amu, and even less than about 500 amu.
[0122] All percentages and ratios used herein, unless otherwise
indicated, are by weight.
[0123] The terms "gene expression" or "protein expression" includes
any information pertaining to the amount of gene transcript or
protein present in a sample, as well as information about the rate
at which genes or proteins are produced or are accumulating or
being degraded (e.g., reporter gene data, data from nuclear runoff
experiments, pulse-chase data etc.). Certain kinds of data might be
viewed as relating to both gene and protein expression. For
example, protein levels in a cell are reflective of the level of
protein as well as the level of transcription, and such data is
intended to be included by the phrase "gene or protein expression
information". Such information may be given in the form of amounts
per cell, amounts relative to a control gene or protein, in
unitless measures, etc.; the term "information" is not to be
limited to any particular means of representation and is intended
to mean any representation that provides relevant information. The
term "expression levels" refers to a quantity reflected in or
derivable from the gene or protein expression data, whether the
data is directed to gene transcript accumulation or protein
accumulation or protein synthesis rates, etc.
[0124] The terms "healthy" and "normal" are used interchangeably
herein to refer to a subject or particular cell or tissue that is
devoid (at least to the limit of detection) of a disease
condition.
[0125] The term "nucleic acid" refers to polynucleotides such as
deoxyribonucleic acid (DNA), and, where appropriate, ribonucleic
acid (RNA). The term should also be understood to include analogues
of either RNA or DNA made from nucleotide analogues, and, as
applicable to the embodiment being described, single-stranded (such
as sense or antisense) and double-stranded polynucleotides. In some
embodiments, "nucleic acid" refers to inhibitory nucleic acids.
Some categories of inhibitory nucleic acid compounds include
antisense nucleic acids, RNAi constructs, and catalytic nucleic
acid constructs. Such categories of nucleic acids are well-known in
the art.
[0126] Embodiments described herein relate to compositions and
methods for modulating hematopoietic stem cells and hematopoiesis.
As described in the Examples below, it was found that that
inhibitors of short-chain dehydrogenase activity, such as 15-PGDH
inhibitors, can be administered to a subject in need thereof, cells
of the subject, and/or tissue of the subject alone or in
combination with a cytokine to increase and/or mobilize
hematopoietic stem cells and/or neutrophils in the blood, marrow,
and/or tissue of the subject.
[0127] In some embodiments, the 15-PGDH inhibitor can be
administered to the subject or a preparation of hematopoietic stem
cells, such as peripheral blood hematopoietic stem cells or
umbilical cord stem cells of the subject, to increase the fitness
of the stem cell preparation as a donor graft or to decrease the
number of units of umbilical cord blood required for
transplantation.
[0128] Hematopoietic stem cells are multipotent stem cells that
give rise to all the blood cell types of an organism, including
myeloid (e.g., monocytes and macrophages, neutrophils, basophils,
eosinophils, erythrocytes, megakaryocytes/platelets, dendritic
cells), and lymphoid lineages (e.g., T-cells, B-cells, NK-cells),
and others known in the art (See Fei, R., et al, U.S. Pat. No.
5,635,387; McGlave, et al, U.S. Pat. No. 5,460,964; Simmons, P., et
al, U.S. Pat. No. 5,677,136; Tsukamoto, et al, U.S. Pat. No.
5,750,397; Schwartz, et al, U.S. Pat. No. 5,759,793; DiGuisto, et
al, U.S. Pat. No. 5,681,599; Tsukamoto, et al, U.S. Pat. No.
5,716,827). Hematopoietic stem cells (HSCs) give rise to committed
hematopoietic progenitor cells (HPCs) that are capable of
generating the entire repertoire of mature blood cells over the
lifetime of an organism.
[0129] Hematopoietic stem cells and hematopoietic progenitor cells
are described herein generally as hematopoietic stem cells unless
noted otherwise and can refer to cells or populations identified by
the presence of the antigenic marker CD34 (CD34+). In some
embodiments, the hematopoietic stem cells can be identified by the
presence of the antigenic marker CD34 and the absence of lineage
(lin) markers and are therefore characterized as
CD34.sup.+/lin.sup.- cells.
[0130] The hematopoietic stem cells used in the methods described
herein may be obtained from any suitable source of hematopoietic
stem and progenitor cells and can be provided as a high purified
population of hematopoietic stem cells or as composition that
includes about 0.01% to about 100% of hematopoietic stem cells. For
example, hematopoietic stem cells may be provided in compositions,
such as unfractionated bone marrow (where the hematopoietic stem
cells comprise less than about 1% of the bone marrow cell
population), umbilical cord blood, placental blood, placenta, fetal
blood, fetal liver, fetal spleen, Wharton's jelly, or mobilized
peripheral blood.
[0131] Suitable sources of hematopoietic stem cells can be isolated
or obtained from an organ of the body containing cells of
hematopoietic origin. The isolated cells can include cells that are
removed from their original environment. For example, a cell is
isolated if it is separated from some or all of the components that
normally accompany it in its native state. For example, an
"isolated population of cells," an "isolated source of cells," or
"isolated hematopoietic stem cells" and the like, as used herein,
refer to in vitro or ex vivo separation of one or more cells from
their natural cellular environment, and from association with other
components of the tissue or organ, i.e., it is not significantly
associated with in vivo substances.
[0132] Hematopoietic stem cells can be obtained or isolated from
bone marrow of adults, which includes femurs, hip, ribs, sternum,
and other bones. Bone marrow aspirates containing hematopoietic
stem cells can be obtained or isolated directly from the hip using
a needle and syringe. Other sources of hematopoietic stem cells
include umbilical cord blood, placental blood, mobilized peripheral
blood, Wharton's jelly, placenta, fetal blood, fetal liver, or
fetal spleen. In particular embodiments, harvesting a sufficient
quantity of hematopoietic stem cells for use in therapeutic
applications may require mobilizing the stem and progenitor cells
in the donor.
[0133] "Hematopoietic stem cell mobilization" refers to the release
of stem cells from the bone marrow into the peripheral blood
circulation for the purpose of leukapheresis, prior to stem cell
transplantation. By increasing the number of stem cells harvested
from the donor, the number of stem cells available for therapeutic
applications can be significantly improved. Hematopoietic growth
factors, e.g., granulocyte colony stimulating factor (G-CSF) or
chemotherapeutic agents often are used to stimulate the
mobilization. Commercial cell mobilization agents exist and can be
used in combination with G-CSF to mobilize sufficient quantities of
hematopoietic stem and progenitor cells for transplantation into a
subject. For example, G-CSF and MOZOBIL (Plerixafor, Genzyme
Corporation) can be administered to a donor in order to harvest a
sufficient number of hematopoietic cells for transplantation. Other
methods of mobilizing hematopoietic stem cells would be apparent to
one having skill in the art.
[0134] In some embodiments, hematopoietic stem cells are obtained
from umbilical cord blood. Cord blood can be harvested according to
techniques known in the art (see, e.g., U.S. Pat. Nos. 7,147,626
and 7,131,958, herein incorporated by reference for such
methodologies).
[0135] In one embodiment, hematopoietic stem cells can be obtained
from pluripotent stem cell sources, e.g., induced pluripotent stem
cells (iPSCs) and embryonic stem cells (ESCs). As used herein, the
term "induced pluripotent stem cell" or "iPSC" refers to a
non-pluripotent cell that has been reprogrammed to a pluripotent
state. Once the cells of a subject have been reprogrammed to a
pluripotent state, the cells can then be programmed to a desired
cell type, such as a hematopoietic stem or progenitor cell. As used
herein, the term "reprogramming" refers to a method of increasing
the potency of a cell to a less differentiated state. As used
herein, the term "programming" refers to a method of decreasing the
potency of a cell or differentiating the cell to a more
differentiated state.
[0136] In some embodiments, the 15-PGDH inhibitor can be
administered to the subject or to hematopoietic stem cells, which
can be administered or contacted ex vivo with one or more 15-PGDH
inhibitors described herein to provide a therapeutic composition.
In one embodiment, the therapeutic compositions or preparations can
include the 15-PGDH inhibitor or a population of hematopoietic stem
cells treated ex vivo with a one or more 15-PGDH inhibitor. In
certain embodiments, the therapeutic composition including the
enhanced hematopoietic stem cells can include whole bone marrow,
umbilical cord blood, or mobilized peripheral blood.
[0137] In particular embodiments, the therapeutic composition
includes a population of cells, wherein the population of cells is
about 95% to about 100% hematopoietic stem cells. The use of
therapeutic compositions of highly purified hematopoietic stem
cells, e.g., a composition comprising a population of cells wherein
the cells comprise about 95% hematopoietic stem cells, may improve
the efficiency of stem cell therapies. Currently practiced methods
of transplantations typically use unfractionated mixtures of cells
where hematopoietic stem cells comprise less than 1% of the total
cell population.
[0138] In some embodiments, the therapeutic composition comprises a
population of cells, wherein the population of cells comprises less
than about 0.1%, 0.5%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, or 30%
hematopoietic stem cells. The population of cells in other
embodiments comprises more than about 0.1%, 0.5%, 1%, 2%, 5%, 10%,
15%, 20%, 25%, or 30% hematopoietic stem cells. In still other
embodiments, the population of cells is about 0.1% to about 1%,
about 1% to about 3%, about 3% to about 5%, about 10%-15%, about
15%-20%, about 20%-25%, about 25%-30%, about 30%-35%, about
35%-40%, about 40%-45%, about 45%-50%, about 60%-70%, about
70%-80%, about 80%-90%, about 90%-95%, or about 95% to about 100%
hematopoietic stem cells.
[0139] Hematopoietic stem cells in the therapeutic compositions
described herein can be autologous/autogeneic ("self) or
non-autologous ("non-self," e.g., allogeneic, syngeneic or
xenogeneic) relative to a subject to which the therapeutic
composition is to be administered. "Autologous," as used herein,
refers to cells from the same subject. "Allogeneic," as used
herein, refers to cells of the same species that differ genetically
to the cell in comparison. "Syngeneic," as used herein, refers to
cells of a different subject that are genetically identical to the
cell in comparison. "Xenogeneic," as used herein, refers to cells
of a different species to the cell in comparison.
[0140] Hematopoietic stem cells for use in the methods described
herein may be depleted of mature hematopoietic cells, such as T
cells, B cells, NK cells, dendritic cells, monocytes, granulocytes,
erythroid cells, and their committed precursors from bone marrow
aspirate, umbilical cord blood, or mobilized peripheral blood
(mobilized leukapheresis product). Mature, lineage committed cells
are depleted by immunodepletion, for example, by labeling solid
substrates with antibodies that bind to a panel of so-called
"lineage" antigens: CD2, CD3, CD11b, CD14, CD15, CD16, CD79, CD56,
CD123, and CD235a. A subsequent step can be performed to further
purify the population of cells, in which a substrate labeled with
antibodies that bind to the CD34.sup.+ antigen are used to isolate
primitive hematopoietic stem cells. Kits are commercially available
for purifying stem and progenitor cells from various cell sources
and in particular embodiments, these kits are suitable for use with
the methods described herein.
[0141] In one embodiment, the amount of hematopoietic stem cells in
the therapeutic composition is at least 0.1.times.10.sup.5 cells,
at least 0.5.times.10.sup.5 cells, at least 1.times.10.sup.5 cells,
at least 5.times.10.sup.5 cells, at least 10.times.10.sup.5 cells,
at least 0.5.times.10.sup.6 cells, at least 0.75.times.10.sup.6
cells, at least 1.times.10.sup.6 cells, at least
1.25.times.10.sup.6 cells, at least 1.5.times.10.sup.6 cells, at
least 1.75.times.10.sup.6 cells, at least 2.times.10.sup.6 cells,
at least 2.5.times.10.sup.6 cells, at least 3.times.10.sup.6 cells,
at least 4.times.10.sup.6 cells, at least 5.times.10.sup.6 cells,
at least 10.times.10.sup.6 cells, at least 15.times.10.sup.6 cells,
at least 20.times.10.sup.6 cells, at least 25.times.10.sup.6 cells,
or at least 30.times.10.sup.6 cells.
[0142] In one embodiment, the amount of hematopoietic stem cells in
the therapeutic composition is the amount of hematopoietic stem
cells in a partial or single cord of blood, or is at least
0.1.times.10.sup.5 cells/kg of bodyweight, at least
0.5.times.10.sup.5 cells/kg of bodyweight, at least
1.times.10.sup.5 cells/kg of bodyweight, at least 5.times.10.sup.5
cells/kg of bodyweight, at least 10.times.10.sup.5 cells/kg of
bodyweight, at least 0.5.times.10.sup.6 cells/kg of bodyweight, at
least 0.75.times.10.sup.6 cells/kg of bodyweight, at least
1.times.10.sup.6 cells/kg of bodyweight, at least
1.25.times.10.sup.6 cells/kg of bodyweight, at least
1.5.times.10.sup.6 cells/kg of bodyweight, at least
1.75.times.10.sup.6 cells/kg of bodyweight, at least
2.times.10.sup.6 cells/kg of bodyweight, at least
2.5.times.10.sup.6 cells/kg of bodyweight, at least
3.times.10.sup.6 cells/kg of bodyweight, at least 4.times.10.sup.6
cells/kg of bodyweight, at least 5.times.10.sup.6 cells/kg of
bodyweight, at least 10.times.10.sup.6 cells/kg of bodyweight, at
least 15.times.10.sup.6 cells/kg of bodyweight, at least
20.times.10.sup.6 cells/kg of bodyweight, at least
25.times.10.sup.6 cells/kg of bodyweight, or at least
30.times.10.sup.6 cells/kg of bodyweight.
[0143] Preparations of the 15-PGDH inhibitor as well as
hematopoietic stem cells administered one or more 15-PGDH
inhibitors and/or therapeutic compositions that include
hematopoietic stem cells and one or more 15-PGDH inhibitor can be
used for improving hematopoietic stem cell transplants and in
treating ischemia or ischemia-damaged tissue, and in reducing
further damage to ischemic tissue and/or repairing damage to
ischemic tissue through cell recruitment, improving vascularization
in ischemic tissue, improving tissue regeneration at sites of
ischemia, decreasing ischemic tissue necrosis or apoptosis, and/or
increasing cell survival at sites of ischemia. In particular
embodiments, the preparations of 15-PGDH inhibitor, 15-PGDH
inhibitor treated hematopoietic stem cells, and/or therapeutic
compositions of 15-PGDH inhibitors and hematopoietic stem cells are
useful to subjects in need of hematopoietic reconstitution, such as
subjects that have undergone or are scheduled to undergo
myeloablative therapy.
[0144] Subjects, who can be treated with the preparations of
15-PGDH inhibitor, 15-PGDH inhibitor treated hematopoietic stem
cells and/or therapeutic compositions of 15-PGDH inhibitors and
hematopoietic stem cells, can include subjects that have or that
have been diagnosed with various types of leukemias, anemias,
lymphomas, myelomas, immune deficiency disorders, and solid tumors.
A subject also includes a human who is a candidate for stem cell
transplant or bone marrow transplantation, such as during the
course of treatment for a malignant disease or a component of gene
therapy. Subjects may also include individuals or animals that
donate stem cells or bone marrow for allogeneic transplantation. In
certain embodiments, a subject may have undergone myeloablative
irradiation therapy or chemotherapy, or may have experienced an
acute radiation or chemical insult resulting in myeloablation. In
certain embodiments, a subject may have undergone irradiation
therapy or chemotherapy, such as during various cancer treatments.
Typical subjects include animals that exhibit aberrant amounts
(lower or higher amounts than a "normal" or "healthy" subject) of
one or more physiological activities that can be modulated by an
agent or a stem cell or marrow transplant.
[0145] Subjects, which can be treated with the preparations of
15-PGDH inhibitors, 15-PGDH inhibitor treated hematopoietic stem
cells and/or therapeutic compositions of 15-PGDH inhibitors and
hematopoietic stem cells, can also include subjects undergoing
chemotherapy or radiation therapy for cancer, as well as subjects
suffering from (e.g., afflicted with) non malignant blood
disorders, particularly immunodeficiencies (e.g. SCID, Fanconi's
anemia, severe aplastic anemia, or congenital hemoglobinopathies,
or metabolic storage diseases, such as Hurler's disease, Hunter's
disease, mannosidosis, among others) or cancer, particularly
hematological malignancies, such as acute leukemia, chronic
leukemia (myeloid or lymphoid), lymphoma (Hodgkin's or
non-Hodgkin's), multiple myeloma, myelodysplastic syndrome, or
non-hematological cancers, such as solid tumors (including breast
cancer, ovarian cancer, brain cancer, prostate cancer, lung cancer,
colon cancer, skin cancer, liver cancer, or pancreatic cancer).
[0146] Subjects may also include subjects suffering from aplastic
anemia, an immune disorder (severe combined immune deficiency
syndrome or lupus), myelodysplasia, thalassemaia, sickle-cell
disease or Wiskott-Aldrich syndrome. In some embodiments, the
subject suffers from a disorder that is the result of an undesired
side effect or complication of another primary treatment, such as
radiation therapy, chemotherapy, or treatment with a bone marrow
suppressive drug, such as zidovadine, chloramphenical or
gangciclovir. Such disorders include neutropenias, anemias,
thrombocytopenia, and immune dysfunction. Other subjects may have
disorders caused by an infection (e.g., viral infection, bacterial
infection or fungal infection) which causes damage to stem or
progenitor cells of the bone marrow.
[0147] In addition, subjects suffering from the following
conditions can also benefit from treatment using the preparations
of 15-PGDH inhibitor, 15-PGDH inhibitor treated hematopoietic stem
cells and/or therapeutic compositions of 15-PGDH inhibitors and
hematopoietic stem cells: lymphocytopenia, lymphorrhea,
lymphostasis, erythrocytopenia, erthrodegenerative disorders,
erythroblastopenia, leukoerythroblastosis; erythroclasis,
thalassemia, myelodysplasia, myelofibrosis, thrombocytopenia,
disseminated intravascular coagulation (DIC), immune (autoimmune)
thrombocytopenic purpura (ITP), HIV inducted ITP, myelodysplasia;
thrombocytotic disease, thrombocytosis, congenital neutropenias
(such as Kostmann's syndrome and Schwachman-Diamond syndrome),
neoplastic associated neutropenias, childhood and adult cyclic
neutropaenia; post-infective neutropaenia; myelodysplastic
syndrome; neutropaenia associated with chemotherapy and
radiotherapy; chronic granulomatous disease; mucopolysaccharidoses;
Diamond Blackfan Anemia; Sickle cell disease; or Beta thalassemia
major.
[0148] In other embodiments, the preparations of 15-PGDH inhibitor,
15-PGDH inhibitor treated hematopoietic stem cells, and/or
therapeutic compositions or 15-PGDH inhibitors and hematopoietic
stem cells can be used in cell-based therapy for treating ischemic
tissue or treating or ameliorating one or more symptoms associated
with tissue ischemia, including, but not limited to, impaired, or
loss of, organ function (including without limitation impairments
or loss of brain, kidney, or heart function), cramping,
claudication, numbness, tingling, weakness, pain, reduced wound
healing, inflammation, skin discoloration, and gangrene.
[0149] In one embodiment, the subject exhibits at least one symptom
of an ischemic tissue or tissue damaged by ischemia. In particular
embodiments, the subject is a human who is has or who is at risk of
having an ischemic tissue or tissue damaged by ischemia, e.g., a
subject that has diabetes, peripheral vascular disease,
thromboangiitis obliterans, vasculitis, cardiovascular disease,
coronary artery disease or heart failure, or cerebrovascular
disease, cardiovascular disease, or cerebrovascular disease.
[0150] Illustrative examples of genetic disorders, syndromic
conditions, traumatic injuries, chronic conditions, medical
interventions, or other conditions that cause or are associated
with ischemia, or increase the risk of ischemia in a subject, or
cause a subject to exhibit more or more symptoms of ischemia, and
thus, suitable for treatment or amelioration using the methods
described herein, include, but are not limited to, acute coronary
syndrome, acute lung injury (ALI), acute myocardial infarction
(AMI), acute respiratory distress syndrome (ARDS), arterial
occlusive disease, arteriosclerosis, articular cartilage defect,
aseptic systemic inflammation, atherosclerotic cardiovascular
disease, autoimmune disease, bone fracture, bone fracture, brain
edema, brain hypoperfusion, Buerger's disease, bums, cancer,
cardiovascular disease, cartilage damage, cerebral infarct,
cerebral ischemia, cerebral stroke, cerebrovascular disease,
chemotherapy-induced neuropathy, chronic infection, chronic
mesenteric ischemia, claudication, congestive heart failure,
connective tissue damage, contusion, coronary artery disease (CAD),
critical limb ischemia (CLI), Crohn's disease, deep vein
thrombosis, deep wound, delayed ulcer healing, delayed
wound-healing, diabetes (type I and type II), diabetic neuropathy,
diabetes induced ischemia, disseminated intravascular coagulation
(DIC), embolic brain ischemia, graft-versus-host disease,
frostbite, hereditary hemorrhagic telengiectasiaischemic vascular
disease, hyperoxic injury, hypoxia, inflammation, inflammatory
bowel disease, inflammatory disease, injured tendons, intermittent
claudication, intestinal ischemia, ischemia, ischemic brain
disease, ischemic heart disease, ischemic peripheral vascular
disease, ischemic placenta, ischemic renal disease, ischemic
vascular disease, ischemic-reperfusion injury, laceration, left
main coronary artery disease, limb ischemia, lower extremity
ischemia, myocardial infarction, myocardial ischemia, organ
ischemia, osteoarthritis, osteoporosis, osteosarcoma, Parkinson's
disease, peripheral arterial disease (PAD), peripheral artery
disease, peripheral ischemia, peripheral neuropathy, peripheral
vascular disease, pre-cancer, pulmonary edema, pulmonary embolism,
remodeling disorder, renal ischemia, retinal ischemia, retinopathy,
sepsis, skin ulcers, solid organ transplantation, spinal cord
injury, stroke, subchondral-bone cyst, thrombosis, thrombotic brain
ischemia, tissue ischemia, transient ischemic attack (TIA),
traumatic brain injury, ulcerative colitis, vascular disease of the
kidney, vascular inflammatory conditions, von Hippel-Lindau
syndrome, and wounds to tissues or organs.
[0151] Other illustrative examples of genetic disorders, syndromic
conditions, traumatic injuries, chronic conditions, medical
interventions, or other conditions that cause or are associated
with ischemia, or increase the risk of ischemia in a subject, or
cause a subject to exhibit more or more symptoms of ischemia
suitable for treatment or amelioration using the methods of the
present invention, include, ischemia resulting from surgery,
chemotherapy, radiation therapy, or cell, tissue, or organ
transplant or graft.
[0152] In various embodiments, the methods are suitable for
treating cerebrovascular ischemia, myocardial ischemia, limb
ischemia (CLI), myocardial ischemia (especially chronic myocardial
ischemia), ischemic cardiomyopathy, cerebrovascular ischemia, renal
ischemia, pulmonary ischemia, intestinal ischemia, and the
like.
[0153] In various embodiments, the preparations or therapeutic cell
compositions disclosed herein can be used to treat an ischemic
tissue in which it is desirable to increase the blood flow, oxygen
supply, glucose supply, or supply of nutrients to the tissue.
[0154] In some embodiments, the 15-PGDH inhibitor can be
administered to a preparation of tissue stem cells, such as neural
stem stems, mesenchymal stem cells, or stem cells that can generate
other tissues, and/or a preparation of pluripotent stem cells.
[0155] In other embodiments, the 15-PGDH inhibitor can be
administered to a bone marrow graft donor or a hematopoietic stem
cell donor to increase the fitness of a donor bone marrow graft or
a donor hematopoietic stem cell graft.
[0156] In other embodiments, the 15-PGDH inhibitor can also be
administered to bone marrow of a subject to increase stem cells in
the subject or to increase the fitness of the marrow as a donor
graft.
[0157] In yet other embodiments, the 15-PGDH inhibitor can be
administered to a subject to mitigate bone marrow graft rejection,
to enhance bone marrow graft engraftment, to enhance engraftment of
a hematopoietic stem cell graft, or an umbilical cord blood stem
cell graft, to enhance engraftment of a hematopoietic stem cell
graft, or an umbilical cord stem cell graft, and/or to decrease the
number of units of umbilical cord blood required for
transplantation into the subject. The administration can be, for
example, following treatment of the subject or the marrow of the
subject with radiation therapy, chemotherapy, or immunosuppressive
therapy.
[0158] In other embodiments, the 15-PGDH inhibitor can be
administered to a recipient of a bone marrow transplant, of a
hematopoietic stem cell transplant, or of an umbilical cord blood
stem cell transplant, in order to decrease the administration of
other treatments or growth factors.
[0159] In some embodiments, the 15-PGDH inhibitor can be
administered to a subject to enhance recovery of neutrophils
following bone marrow transplantation, following umbilical cord
blood transplantation, following transplantation with hematopoietic
stem cells, following conventional chemotherapy, following
radiation treatment, and in individuals with neutropenias from
diseases that include but are not limited to aplastic anemia,
myelodysplasia, myelofibrosis, neutropenias from other bone marrow
diseases, drug induced neutropenia, immune neutropenias, idiopathic
neutropenia, and following infections with viruses that include,
but are not limited to, HIV, CMV, and parvovirus.
[0160] In other embodiments, the 15-PGDH inhibitor can be
administered to a subject to enhance recovery of platelets
following bone marrow transplantation, following umbilical cord
blood transplantation, following transplantation with hematopoietic
stem cells, following conventional chemotherapy, following
radiation treatment, and in individuals with neutropenias from
diseases that include but are not limited to aplastic anemia,
myelodysplasia, myelofibrosis, thrombocytopenias from other bone
marrow diseases, drug induced thrombocytopenia, immune
thrombocytopenia, idiopathic thrombocytopenic purpura, idiopathic
thrombocytopenia, and following infections with viruses that
include, but are not limited to, HIV, CMV, and parvovirus.
[0161] In still other embodiments, the 15-PGDH inhibitor can be
administered to a subject to enhance recovery of hemoglobin
following bone marrow transplantation, following umbilical cord
blood transplantation, following transplantation with hematopoietic
stem cells, following conventional chemotherapy, following
radiation treatment, and in individuals with anemias from diseases
that include but are not limited to aplastic anemia,
myelodysplasia, myelofibrosis, anemia from other bone marrow
diseases, drug induced anemia, immune mediated anemias, anemia of
chronic disease, idiopathic anemia, and following infections with
viruses that include, but are not limited to, HIV, CMV, and
parvovirus.
[0162] In some embodiments, the 15-PGDH inhibitor can be
administered to a subject to enhance numbers of bone marrow stem
cell numbers following bone marrow transplantation, following
umbilical cord blood transplantation, following transplantation
with hematopoietic stem cells, following conventional chemotherapy,
following radiation treatment, in individuals with other bone
marrow diseases, in individuals with cytopenias following viral
infections, and in individuals with cytopenias.
[0163] In other embodiments, the 15-PGDH inhibitor can be
administered to a subject to enhance response to cytokines and/or
cell mobilization agents administered to individuals with
cytopenias that include but are not limited to neutropenia,
thrombocytopenia, lymphocytopenia, and anemia. Cytokines whose
responses may be enhanced by 15-PGDH inhibitors include, but are
not limited to: G-CSF, GM-CSF, EPO, IL-3, IL-6, TPO, SCF, and
TPO-RA (thrombopoietin receptor agonist).
[0164] In further embodiments, the 15-PGDH inhibitor can be
administered to a subject or to a tissue graft of a subject to
mitigate graft rejection, to enhance graft engraftment, to enhance
graft engraftment following treatment of the subject or the marrow
of the subject with radiation therapy, chemotherapy, or
immunosuppressive therapy, to confer resistance to toxic or lethal
effects of exposure to radiation, confer resistance to the toxic
effect of Cytoxan, the toxic effect of fludarabine, the toxic
effect of chemotherapy, or the toxic effect of immunosuppressive
therapy, to decrease infection, and/or to decrease pulmonary
toxicity from radiation.
[0165] In other embodiments, the 15-PGDH inhibitor can be
administered to a recipient of a tissue stem cell transplant,
including but not limited to a transplant with hematopoietic stem
cells, neural stem stems, mesenchymal stem cells, or stem cells for
other tissues, so as to accelerate tissue regeneration and repair
following the transplant.
[0166] In some embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing neutrophils in the subject. For
example, the hematopoietic cytokine or cell mobilization agent can
include at least one of G-CSF or Plerixafor.
[0167] In other embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing numbers of and/or of mobilizing
peripheral blood hematopoietic stem cells in the subject. For
example, the hematopoietic cytokine or cell mobilization agent can
include at least one of G-CSF or Plerixafor.
[0168] In still other embodiments, the administration of a 15-PGDH
inhibitor alone or in combination with a cytokine, hematopoietic
cytokine, and/or cell mobilization agent to a subject in need
thereof, cells of the subject, and/or tissue of the subject can be
used for the purpose of increasing numbers of hematopoietic stem
cells in blood or bone marrow of the subject. For example, the
hematopoietic cytokine or cell mobilization agent can include at
least one of G-CSF or Plerixafor.
[0169] 15-PGDH inhibitors can be identified using assays in which
putative inhibitor compounds are applied to cells expressing
15-PGDH and then the functional effects on 15-PGDH activity are
determined. Samples or assays comprising 15-PGDH that are treated
with a potential inhibitor are compared to control samples without
the inhibitor to examine the extent of effect. Control samples
(untreated with modulators) are assigned a relative 15-PGDH
activity value of 100%. Inhibition of 15-PGDH is achieved when the
15-PGDH activity value relative to the control is about 80%,
optionally 50% or 25%, 10%, 5% or 1%.
[0170] Agents tested as inhibitors of SCD (e.g., 15-PGDH) can be
any small chemical molecule or compound. Typically, test compounds
will be small chemical molecules, natural products, or peptides.
The assays are designed to screen large chemical libraries by
automating the assay steps and providing compounds from any
convenient source to assays, which are typically run in parallel
(e.g., in microtiter formats on microtiter plates in robotic
assays).
[0171] In some embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (I):
##STR00008## [0172] wherein n is 0-2; [0173] Y.sup.1, Y.sup.2, and
R.sup.1 are the same or different and are each selected from the
group consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O) (C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH2, --SO.sub.2NY.sub.2
(wherein Y is independently H, arlyl or alkyl), phosphono
(--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein Y.sup.1 and Y.sup.2 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; [0174] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0175]
X.sup.1 and X.sup.2 are independently N or C, and wherein when
X.sup.1 and/or X.sup.2 are N, Y and/or Y.sup.2, respectively, are
absent; [0176] Z.sup.1 is O, S, CR.sup.aR.sup.b or NR.sup.a,
wherein R.sup.a and R.sup.b are independently H or a C.sub.1-8
alkyl, which is linear, branched, or cyclic, and which is
unsubstituted or substituted; [0177] and pharmaceutically
acceptable salts thereof.
[0178] Examples of 15-PGDH inhibitors having formulas (I) include
the following compounds:
##STR00009##
and pharmaceutically acceptable salts thereof.
[0179] In other embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (II):
##STR00010## [0180] wherein n is 0-2 [0181] X.sup.4, X.sup.5,
X.sup.6, and X.sup.7 are independently N or CR.sup.c; [0182]
R.sup.1, R.sup.6, R.sup.7, and R.sup.c are independently selected
from the group consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O)(C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH2, --SO.sub.2NY.sub.2
(wherein Y is independently H, aryl or alkyl), phosphono
(--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; [0183] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0184]
Z.sup.1 is O, S, CR.sup.aR.sup.b or NR.sup.a, wherein R.sup.a and
R.sup.b are independently H or a C.sub.1-8 alkyl, which is linear,
branched, or cyclic, and which is unsubstituted or substituted;
[0185] and pharmaceutically acceptable salts thereof.
[0186] Examples of 15-PGDH inhibitors having formulas (II) include
the following compounds:
##STR00011##
and pharmaceutically acceptable salts thereof.
[0187] In yet other embodiments, the 15-PGDH inhibitor can include
a compound having the following formula (III) or (IV):
##STR00012## [0188] wherein n is 0-2 [0189] X.sup.6 is
independently is N or CR.sup.c; [0190] R.sup.1, R.sup.6, R.sup.7,
and R.sup.c are independently selected from the group consisting of
hydrogen, substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl containing
from 5-6 ring atoms (wherein from 1-3 of the ring atoms is
independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O)(C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato (--SO.sub.2--O),
C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl; also termed
"alkylthio"), arylsulfanyl (--S-aryl; also termed "arylthio"),
C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl), C.sub.5-C.sub.20
arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24 alkylsulfonyl
(--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; [0191] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0192]
Z.sup.1 is O, S, CR.sup.aR.sup.b or NR.sup.a, wherein R.sup.a and
R.sup.b are independently H or a C.sub.1-8 alkyl, which is linear,
branched, or cyclic, and which is unsubstituted or substituted;
[0193] and pharmaceutically acceptable salts thereof.
[0194] In some embodiments, R.sup.1 is selected from the group
consisting of branched or linear alkyl including
--(CH.sub.2).sub.n1CH.sub.3 (n.sub.1=0-7),
##STR00013##
wherein n.sub.2=0-6 and X is any of the following: CF.sub.yHz
(y+z=3), CCl.sub.yHz (y+z=3), OH, OAc, OMe, R.sup.71, OR.sup.72,
CN, N(R.sup.73).sub.2,
##STR00014##
(n.sub.3=0-5, m=1-5), and
##STR00015##
(n.sub.4=0-5).
[0195] In other embodiments, R.sup.6 and R.sup.7 can each
independently be one of the following:
##STR00016## ##STR00017## [0196] each R.sup.8, R.sup.9, R.sup.10,
R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16,
R.sup.17, R.sup.18, R.sup.19, R.sup.20, R.sup.21, R.sup.22,
R.sup.23, R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28,
R.sup.29, R.sup.30, R.sup.31, R.sup.32, R.sup.33, R.sup.34,
R.sup.35, R.sup.36, R.sup.37, R.sup.38, R.sup.39, R.sup.40,
R.sup.41, R.sup.42, R.sup.43, R.sup.44, R.sup.45, R.sup.46,
R.sup.47, R.sup.48, R.sup.49, R.sup.50, R.sup.51, R.sup.52,
R.sup.53, R.sup.54, R.sup.55, R.sup.56, R.sup.57, R.sup.58,
R.sup.59, R.sup.60, R.sup.61, R.sup.62, R.sup.63, R.sup.64,
R.sup.65, R.sup.66, R.sup.67, R.sup.68, R.sup.69, R.sup.70,
R.sup.71, R.sup.72, R.sup.73 and R.sup.74 are the same or different
and are independently selected from the group consisting of
hydrogen, substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heterocycloalkenyl containing from 5-6 ring
atoms, (wherein from 1-3 of the ring atoms is independently
selected from N, NH, N(C.sub.1-C.sub.6 alkyl), NC(O)
(C.sub.1-C.sub.6 alkyl), O, and S), heteroaryl or heterocyclyl
containing from 5-14 ring atoms, (wherein from 1-6 of the ring
atoms is independently selected from N, NH, N(C.sub.1-C.sub.3
alkyl), O, and S), C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, silyl, hydroxyl, sulfhydryl, C.sub.1-C.sub.24
alkoxy, C.sub.2-C.sub.24 alkenyloxy, C.sub.2-C.sub.24 alkynyloxy,
C.sub.5-C.sub.20 aryloxy, acyl (including C.sub.2-C.sub.24
alkylcarbonyl (--CO-alkyl) and C.sub.6-C.sub.20 arylcarbonyl
(--CO-aryl)), acyloxy (--O-acyl), C.sub.2-C.sub.24 alkoxycarbonyl
(--(CO)--O-alkyl), C.sub.6-C.sub.20 aryloxycarbonyl
(--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), sulfanamido (--SO.sub.2N(R).sub.2
where R is independently H, alkyl, aryl or heteroaryl), imino
(--CR.dbd.NH where R is hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl),
where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkyl ethers (--[(CH.sub.2).sub.nO].sub.m),
phosphates, phosphate esters [--OP(O)(OR).sub.2 where R.dbd.H,
methyl or other alkyl], groups incorporating amino acids or other
moieties expected to bear positive or negative charge at
physiological pH, and combinations thereof, and pharmaceutically
acceptable salts thereof.
[0197] In still other embodiments, R.sup.6 and R.sup.7 can
independently be a group that improves aqueous solubility, for
example, a phosphate ester (--OPO.sub.3H.sub.2), a phenyl ring
linked to a phosphate ester (--OPO.sub.3H.sub.2), a phenyl ring
substituted with one or more methoxyethoxy groups, or a morpholine,
or an aryl or heteroaryl ring substituted with such a group.
[0198] Examples of 15-PGDH inhibitors having formulas (III) or (IV)
include the following compounds:
##STR00018## ##STR00019## ##STR00020##
and pharmaceutically acceptable salts thereof.
[0199] In other embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (V):
##STR00021## [0200] wherein n is 0-2 [0201] X.sup.6 is
independently is N or CR.sup.c [0202] R.sup.1, R.sup.6, R.sup.7,
and R.sup.c are each independently selected from the group
consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O)(C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH2, --SO.sub.2NY.sub.2
(wherein Y is independently H, aryl or alkyl), phosphono
(--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; [0203] U.sup.1 is N, C--R.sup.2, or
C--NR.sup.3R.sup.4, wherein R.sup.2 is selected from the group
consisting of a H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, CH.sub.2--CH.sub.2--CH.sub.2X,
O--CH.sub.2--CH.sub.2X, X, (wherein X.dbd.H, F, Cl, Br, or I), CN,
(C.dbd.O)--R', (C.dbd.O)N(R').sub.2, O(CO)R', COOR' (wherein R' is
H or a lower alkyl group), and wherein R.sup.1 and R.sup.2 may be
linked to form a cyclic or polycyclic ring, wherein R.sup.3 and
R.sup.4 are the same or different and are each selected from the
group consisting of H, a lower alkyl group, O, (CH.sub.2).sub.n1OR'
(wherein n1=1, 2, or 3), CF.sub.3, CH.sub.2--CH.sub.2X,
CH.sub.2--CH.sub.2--CH.sub.2X, (wherein X.dbd.H, F, Cl, Br, or I),
CN, (C.dbd.O)--R', (C.dbd.O)N(R').sub.2, COOR' (wherein R' is H or
a lower alkyl group), and R.sup.3 or R.sup.4 may be absent; [0204]
and pharmaceutically acceptable salts thereof.
[0205] In some embodiments, R.sup.1 is selected from the group
consisting of branched or linear alkyl including
--(CH.sub.2)n.sub.1CH.sub.3 (n.sub.1=0-7),
##STR00022##
wherein n.sub.2=0-6 and X is any of the following: CF.sub.yH.sub.z
(y+z=3), CCl.sub.yHz (y+z=3), OH, OAc, OMe, R.sup.7l, OR.sup.72,
CN, N(R.sup.73).sub.2,
##STR00023##
(n.sub.3=0-5, m=1-5), and
##STR00024##
(n.sub.4=0-5).
[0206] In other embodiments, R.sup.6 and R.sup.7 can each
independently be one of the following:
##STR00025## ##STR00026##
[0207] each R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12,
R.sup.13, R.sup.14, R.sup.15, R.sup.16, R.sup.17, R.sup.18,
R.sup.19, R.sup.20, R.sup.21, R.sup.22, R.sup.23, R.sup.24,
R.sup.25, R.sup.26, R.sup.27, R.sup.28, R.sup.29, R.sup.30,
R.sup.31, R.sup.32, R.sup.33, R.sup.34, R.sup.35, R.sup.36,
R.sup.37, R.sup.38, R.sup.39, R.sup.40, R.sup.41, R.sup.42,
R.sup.43, R.sup.44, R.sup.45, R.sup.46, R.sup.47, R.sup.48,
R.sup.49, R.sup.50, R.sup.51, R.sup.52, R.sup.53, R.sup.54,
R.sup.55, R.sup.56, R.sup.57, R.sup.58, R.sup.59, R.sup.60,
R.sup.61, R.sup.62, R.sup.63, R.sup.64, R.sup.65, R.sup.66,
R.sup.67, R.sup.68, R.sup.69, R.sup.70, R.sup.71, R.sup.72,
R.sup.73, and R.sup.74, are the same or different and are
independently selected from the group consisting of hydrogen,
substituted or unsubstituted C.sub.1-C.sub.24 alkyl,
C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24 alkynyl,
C.sub.3-C.sub.20 aryl, heterocycloalkenyl containing from 5-6 ring
atoms, (wherein from 1-3 of the ring atoms is independently
selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O)(C.sub.1-C.sub.6 alkyl), O, and S), heteroaryl or heterocyclyl
containing from 5-14 ring atoms, (wherein from 1-6 of the ring
atoms is independently selected from N, NH, N(C.sub.1-C.sub.3
alkyl), O, and S), C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, silyl, hydroxyl, sulfhydryl, C.sub.1-C.sub.24
alkoxy, C.sub.2-C.sub.24 alkenyloxy, C.sub.2-C.sub.24 alkynyloxy,
C.sub.5-C.sub.20 aryloxy, acyl (including C.sub.2-C.sub.24
alkylcarbonyl (--CO-alkyl) and C.sub.6-C.sub.20 arylcarbonyl
(--CO-aryl)), acyloxy (--O-acyl), C.sub.2-C.sub.24 alkoxycarbonyl
(--(CO)--O-alkyl), C.sub.6-C.sub.20 aryloxycarbonyl
(--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), sulfanamido (--SO.sub.2N(R).sub.2
where R is independently H, alkyl, aryl or heteroaryl), imino
(--CR.dbd.NH where R is hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl),
where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkyl ethers (--[(CH.sub.2).sub.nO].sub.m),
phosphates, phosphate esters [--OP(O)(OR).sub.2 where R.dbd.H,
methyl or other alkyl], groups incorporating amino acids or other
moieties expected to bear positive or negative charge at
physiological pH, and combinations thereof, and pharmaceutically
acceptable salts thereof.
[0208] In still other embodiments, R.sup.6 and R.sup.7 can
independently be a group that improves aqueous solubility, for
example, a phosphate ester (--OPO.sub.3H.sub.2), a phenyl ring
linked to a phosphate ester (--OPO.sub.3H.sub.2), a phenyl ring
substituted with one or more methoxyethoxy groups, or a morpholine,
or an aryl or heteroaryl ring substituted with such a group.
[0209] In other embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (VI):
##STR00027## [0210] wherein n=0-2; [0211] X.sup.6 is N or CR.sup.c;
[0212] R.sup.1 is selected from the group consisting of branched or
linear alkyl including --(CH.sub.2)n.sub.1CH.sub.3
(n.sub.1=0-7),
##STR00028##
[0212] wherein n.sub.2=0-6 and X is any of the following:
CF.sub.yH.sub.z (y+z=3), CCl.sub.yHz (y+z=3), OH, OAc, OMe,
R.sup.71, OR.sup.72, CN, N(R.sup.73).sub.2,
##STR00029##
(n.sub.3=0-5, m=1-5), and
##STR00030##
(n.sub.4=0-5). [0213] R.sup.5 is selected from the group consisting
of H, Cl, F, NH.sub.2, and N(R.sup.76).sub.2; [0214] R.sup.6 and
R.sup.7 can each independently be one of the following:
[0214] ##STR00031## ##STR00032## [0215] each R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15,
R.sup.16, R.sup.17, R.sup.18, R.sup.20, R.sup.21, R.sup.22,
R.sup.23, R.sup.24, R.sup.25, R.sup.26, R.sup.27, R.sup.28,
R.sup.29, R.sup.30, R.sup.31, R.sup.32, R.sup.33, R.sup.34,
R.sup.35, R.sup.36, R.sup.37, R.sup.38, R.sup.39, R.sup.40,
R.sup.41, R.sup.42, R.sup.43, R.sup.44, R.sup.45, R.sup.46,
R.sup.47, R.sup.48, R.sup.49, R.sup.50, R.sup.51, R.sup.52,
R.sup.53, R.sup.54, R.sup.55, R.sup.56, R.sup.57, R.sup.58,
R.sup.59, R.sup.60, R.sup.61, R.sup.62, R.sup.63, R.sup.64,
R.sup.65, R.sup.66, R.sup.67, R.sup.68, R.sup.69, R.sup.70,
R.sup.71, R.sup.72, R.sup.73, R.sup.74, R.sup.76, and R.sup.c are
the same or different and are independently selected from the group
consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heterocycloalkenyl containing from
5-6 ring atoms, (wherein from 1-3 of the ring atoms is
independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl), NC(O)
(C.sub.1-C.sub.6 alkyl), O, and S), heteroaryl or heterocyclyl
containing from 5-14 ring atoms, (wherein from 1-6 of the ring
atoms is independently selected from N, NH, N(C.sub.1-C.sub.3
alkyl), O, and S), C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, halo, silyl, hydroxyl, sulfhydryl, C.sub.1-C.sub.24
alkoxy, C.sub.2-C.sub.24 alkenyloxy, C.sub.2-C.sub.24 alkynyloxy,
C.sub.5-C.sub.20 aryloxy, acyl (including C.sub.2-C.sub.24
alkylcarbonyl (--CO-alkyl) and C.sub.6-C.sub.20 arylcarbonyl
(--CO-aryl)), acyloxy (--O-acyl), C.sub.2-C.sub.24 alkoxycarbonyl
(--(CO)--O-alkyl), C.sub.6-C.sub.20 aryloxycarbonyl
(--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), sulfanamido (--SO.sub.2N(R).sub.2
where R is independently H, alkyl, aryl or heteroaryl), imino
(--CR.dbd.NH where R is hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl),
where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato (--SO.sub.2--O),
C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl; also termed
"alkylthio"), arylsulfanyl (--S-aryl; also termed "arylthio"),
C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl), C.sub.5-C.sub.20
arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24 alkylsulfonyl
(--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkyl ethers (--[(CH.sub.2).sub.nO].sub.m),
phosphates, phosphate esters [--OP(O)(OR).sub.2 where R.dbd.H,
methyl or other alkyl], groups incorporating amino acids or other
moieties expected to bear positive or negative charge at
physiological pH, and combinations thereof, and pharmaceutically
acceptable salts thereof.
[0216] In other embodiments, the 15-PGDH inhibitor can include a
compound having the following formula (VII):
##STR00033## [0217] wherein n=0-2; [0218] X.sup.6 is N or CR.sup.c;
[0219] R.sup.1 is selected from the group consisting of branched or
linear alkyl including --(CH.sub.2)n.sub.1CH.sub.3
(n.sub.1=0-7),
##STR00034##
[0219] wherein n.sub.2=0-6 and X is any of the following:
CF.sub.yH.sub.z (y+z=3), CCl.sub.yH.sub.z (y+z=3), OH, OAc, OMe,
R.sup.71, OR.sup.72, CN, N(R.sup.73).sub.2,
##STR00035##
(n.sub.3=0-5, m=1-5), and
##STR00036##
(n.sub.4=0-5). [0220] R.sup.5 is selected from the group consisting
of H, Cl, F, NH.sub.2, and N(R.sup.76).sub.2; [0221] R.sup.7 can
each independently be one of the following:
[0221] ##STR00037## ##STR00038## [0222] each R.sup.8, R.sup.9,
R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15,
R.sup.16, R.sup.17, R.sup.18, R.sup.19, R.sup.20, R.sup.21,
R.sup.22, R.sup.23, R.sup.24, R.sup.25, R.sup.26, R.sup.27,
R.sup.28, R.sup.29, R.sup.30, R.sup.31, R.sup.32, R.sup.33,
R.sup.34, R.sup.35, R.sup.36, R.sup.37, R.sup.38, R.sup.39,
R.sup.40, R.sup.41, R.sup.42, R.sup.43, R.sup.44, R.sup.45,
R.sup.46, R.sup.47, R.sup.48, R.sup.49, R.sup.50, R.sup.51,
R.sup.52, R.sup.53, R.sup.54, R.sup.55, R.sup.56, R.sup.57,
R.sup.58, R.sup.59, R.sup.60, R.sup.61, R.sup.62, R.sup.63,
R.sup.64, R.sup.65, R.sup.66, R.sup.67, R.sup.68, R.sup.69,
R.sup.70, R.sup.71, R.sup.72, R.sup.73, R.sup.74, R.sup.76, and
R.sup.c are the same or different and are independently selected
from the group consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl,
C.sub.2alkenyloxy, C.sub.6-C.sub.24 alkynyl, C.sub.3-C.sub.2 aryl,
heterocycloalkenyl containing from 5-6 ring atoms, (wherein from
1-3 of the ring atoms is independently selected from N, NH,
N(C.sub.1-C.sub.6 alkyl), NC(O)(C.sub.1-C.sub.6 alkyl), O, and S),
heteroaryl or heterocyclyl containing from 5-14 ring atoms,
(wherein from 1-6 of the ring atoms is independently selected from
N, NH, N(C.sub.1-C.sub.3 alkyl), O, and S), C.sub.6-C.sub.24
alkaryl, C.sub.6-C.sub.24 aralkyl, halo, silyl, hydroxyl,
sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24 alkenyloxy,
C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy, acyl
(including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), sulfanamido (--SO.sub.2N(R).sub.2
where R is independently H, alkyl, aryl or heteroaryl), imino
(--CR.dbd.NH where R is hydrogen, C.sub.1-C.sub.24 alkyl,
C.sub.5-C.sub.20 aryl, C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24
aralkyl, etc.), alkylimino (--CR.dbd.N(alkyl), where R=hydrogen,
alkyl, aryl, alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl),
where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH.sub.2,
--SO.sub.2NY.sub.2 (wherein Y is independently H, aryl or alkyl),
phosphono (--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkyl ethers (--[(CH.sub.2).sub.nO].sub.m),
phosphates, phosphate esters [--OP(O)(OR).sub.2 where R.dbd.H,
methyl or other alkyl], groups incorporating amino acids or other
moieties expected to bear positive or negative charge at
physiological pH, and combinations thereof, and pharmaceutically
acceptable salts thereof.
[0223] Examples of compounds having formulas (V), (VI), or (VII)
are selected from the group consisting of:
##STR00039## ##STR00040## ##STR00041## ##STR00042## ##STR00043##
##STR00044## ##STR00045## ##STR00046## ##STR00047## ##STR00048##
##STR00049## ##STR00050## ##STR00051## ##STR00052## ##STR00053##
##STR00054## ##STR00055## ##STR00056## ##STR00057## ##STR00058##
##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063##
##STR00064## ##STR00065## ##STR00066## ##STR00067## ##STR00068##
##STR00069## ##STR00070## [0224] and pharmaceutically acceptable
salts thereof.
[0225] In certain embodiments, the 15-PGDH inhibitor having formula
(I), (II), (III), (IV), (V), (VI), and (VII) can be selected that
can ia) at 2.5 .mu.M concentration, stimulate a Vaco503 reporter
cell line expressing a 15-PGDH luciferase fusion construct to a
luciferase output level of greater than 70 (using a scale on which
a value of 100 indicates a doubling of reporter output over
baseline); iia) at 2.5 .mu.M concentration stimulate a V9m reporter
cell line expressing a 15-PGDH luciferase fusion construct to a
luciferase output level of greater than 75; iiia) at 7.5 .mu.M
concentration stimulate a LS174T reporter cell line expressing a
15-PGDH luciferase fusion construct to a luciferase output level of
greater than 70; and iva) at 7.5 .mu.M concentration, does not
activate a negative control V9m cell line expressing TK-renilla
luciferase reporter to a level greater than 20; and va) inhibits
the enzymatic activity of recombinant 15-PGDH protein at an
IC.sub.50 of less than 1 .mu.M.
[0226] In other embodiments, the 15-PGDH inhibitor can ib) at 2.5
.mu.M concentration, stimulate a Vaco503 reporter cell line
expressing a 15-PGDH luciferase fusion construct to increase
luciferase output; iib) at 2.5 .mu.M concentration stimulate a V9m
reporter cell line expressing a 15-PGDH luciferase fusion construct
to increase luciferase output; iiib) at 7.5 .mu.M concentration
stimulate a LS174T reporter cell line expressing a 15-PGDH
luciferase fusion construct to increase luciferase output; ivb) at
7.5 .mu.M concentration, does not activate a negative control V9m
cell line expressing TK-renilla luciferase reporter to a luciferase
level greater than 20% above background; and vb) inhibits the
enzymatic activity of recombinant 15-PGDH protein at an IC.sub.50
of less than 1 .mu.M.
[0227] In other embodiments, the 15-PGDH inhibitor can inhibit the
enzymatic activity of recombinant 15-PGDH at an IC.sub.50 of less
than 1 .mu.M, or preferably at an IC.sub.50 of less than 250 nM, or
more preferably at an IC.sub.50 of less than 50 nM, or more
preferably at an IC.sub.50 of less than 10 nM, or more preferably
at an IC.sub.50 of less than 5 nM at a recombinant 15-PGDH
concentration of about 5 nM to about 10 nM.
[0228] In other embodiments, the 15-PGDH inhibitor can increase the
cellular levels of PGE-2 following stimulation of an A459 cell with
an appropriate agent, for example IL1-beta.
[0229] In some embodiments, a15-PGDH inhibitor can include a
compound having the following formula (VIII):
##STR00071## [0230] wherein n is 0-2; [0231] R.sup.1, R.sup.6, and
R.sup.7 are the same or different and are each selected from the
group consisting of hydrogen, substituted or unsubstituted
C.sub.1-C.sub.24 alkyl, C.sub.2-C.sub.24 alkenyl, C.sub.2-C.sub.24
alkynyl, C.sub.3-C.sub.20 aryl, heteroaryl, heterocycloalkenyl
containing from 5-6 ring atoms (wherein from 1-3 of the ring atoms
is independently selected from N, NH, N(C.sub.1-C.sub.6 alkyl),
NC(O) (C.sub.1-C.sub.6 alkyl), O, and S), C.sub.6-C.sub.24 alkaryl,
C.sub.6-C.sub.24 aralkyl, halo, --Si(C.sub.1-C.sub.3 alkyl).sub.3,
hydroxyl, sulfhydryl, C.sub.1-C.sub.24 alkoxy, C.sub.2-C.sub.24
alkenyloxy, C.sub.2-C.sub.24 alkynyloxy, C.sub.5-C.sub.20 aryloxy,
acyl (including C.sub.2-C.sub.24 alkylcarbonyl (--CO-alkyl) and
C.sub.6-C.sub.20 arylcarbonyl (--CO-aryl)), acyloxy (--O-acyl),
C.sub.2-C.sub.24 alkoxycarbonyl (--(CO)--O-alkyl), C.sub.6-C.sub.20
aryloxycarbonyl (--(CO)--O-aryl), C.sub.2-C.sub.24 alkylcarbonato
(--O--(CO)--O-alkyl), C.sub.6-C.sub.20 arylcarbonato
(--O--(CO)--O-aryl), carboxy (--COOH), carboxylato (--COO.sup.-),
carbamoyl (--(CO)--NH.sub.2), C.sub.1-C.sub.24 alkyl-carbamoyl
(--(CO)--NH(C.sub.1-C.sub.24 alkyl)), arylcarbamoyl
(--(CO)--NH-aryl), thiocarbamoyl (--(CS)--NH.sub.2), carbamido
(--NH--(CO)--NH.sub.2), cyano(-CN), isocyano (--N.sup.+C.sup.-),
cyanato (--O--CN), isocyanato (--O--N.sup.+.dbd.C.sup.-),
isothiocyanato (--S--CN), azido (--N.dbd.N.sup.+.dbd.N.sup.-),
formyl (--(CO)--H), thioformyl (--(CS)--H), amino (--NH.sub.2),
C.sub.1-C.sub.24 alkyl amino, C.sub.5-C.sub.20 aryl amino,
C.sub.2-C.sub.24 alkylamido (--NH--(CO)-alkyl), C.sub.6-C.sub.20
arylamido (--NH--(CO)-aryl), imino (--CR.dbd.NH where R is
hydrogen, C.sub.1-C.sub.24 alkyl, C.sub.5-C.sub.20 aryl,
C.sub.6-C.sub.24 alkaryl, C.sub.6-C.sub.24 aralkyl, etc.),
alkylimino (--CR.dbd.N(alkyl), where R=hydrogen, alkyl, aryl,
alkaryl, aralkyl, etc.), arylimino (--CR.dbd.N(aryl), where
R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (--NO.sub.2),
nitroso (--NO), sulfo (--SO.sub.2--OH), sulfonato
(--SO.sub.2--O.sup.-), C.sub.1-C.sub.24 alkylsulfanyl (--S-alkyl;
also termed "alkylthio"), arylsulfanyl (--S-aryl; also termed
"arylthio"), C.sub.1-C.sub.24 alkylsulfinyl (--(SO)-alkyl),
C.sub.5-C.sub.20 arylsulfinyl (--(SO)-aryl), C.sub.1-C.sub.24
alkylsulfonyl (--SO.sub.2-alkyl), C.sub.5-C.sub.20 arylsulfonyl
(--SO.sub.2-aryl), sulfonamide (--SO.sub.2--NH2, --SO.sub.2NY.sub.2
(wherein Y is independently H, aryl or alkyl), phosphono
(--P(O)(OH).sub.2), phosphonato (--P(O)(O.sup.-).sub.2),
phosphinato (--P(O)(O.sup.-)), phospho (--PO.sub.2), phosphino
(--PH.sub.2), polyalkylethers, phosphates, phosphate esters, groups
incorporating amino acids or other moieties expected to bear
positive or negative charge at physiological pH, combinations
thereof, and wherein R.sup.6 and R.sup.7 may be linked to form a
cyclic or polycyclic ring, wherein the ring is a substituted or
unsubstituted aryl, a substituted or unsubstituted heteroaryl, a
substituted or unsubstituted cycloalkyl, and a substituted or
unsubstituted heterocyclyl; and pharmaceutically acceptable salts
thereof.
[0232] 15-PGDH inhibitors having formula (VIII) can be synthesized
as shown:
##STR00072##
[0233] Any reaction solvent can be used in the above preparation
process as long as it is not involved in the reaction. For example,
the reaction solvent includes ethers such as diethyl ether,
tetrahydrofuran and dioxane; halogenized hydrocarbons, such as
dichloromethane and chloroform; amines such as pyridine, piperidine
and triethylamine; alkylketones, such as acetone, methylethylketone
and methylisobutyl; alcohols, such as methanol, ethanol and
propanol; non-protonic polar solvent, such as
N,N-dimethylformamide, N,N-dimethylacetamide, acetonitrile,
dimethylsulfoxide and hexamethyl phosphoric acid triamide. Among
non-reactive organic solvents that are ordinarily used in the
organic synthesis, preferable solvents are those from which water
generated in the reaction can be removed by a Dean-Stark trap. The
examples of such solvents include, but are not limited to benzene,
toluene, xylene and the like. The reaction product thus obtained
may be isolated and purified by condensation, extraction and the
like, which is ordinarily conducted in the field of the organic
synthesis, if desired, by silica gel column chromatography. The
individual enantiomers of PGDH inhibitors having the formula III
can be separated by a preparative HPLC using chromatography columns
containing chiral stationary phases.
[0234] Further, embodiments of this application include any
modifications for the preparation method of the 15-PGDH inhibitors
described above. In this connection, any intermediate product
obtainable from any step of the preparation method can be used as a
starting material in the other steps. Such starting material can be
formed in situ under certain reaction conditions. Reaction reagents
can also be used in the form of their salts or optical isomers.
[0235] Depending on the kinds of the substituents to be used in the
preparation of the 15-PGDH inhibitors, and the intermediate product
and the preparation method selected, novel 15-PGDH inhibitors can
be in the form of any possible isomers such as substantially pure
geometrical (cis or trans) isomers, optical isomers (enantiomers)
and racemates.
[0236] In some embodiments, a 15-PGDH inhibitor having formula
(VIII) can include a compound with the following formula (IX):
##STR00073## [0237] and pharmaceutically acceptable salts
thereof.
[0238] Advantageously, the 15-PDGH inhibitor having formula (IX)
was found to: i) inhibit recombinant 15-PGDH at 1 nM concentration;
ii) inhibit 15-PGDH in cell lines at 100 nM concentration, iii)
increase PGE.sub.2 production by cell lines; iv) is chemically
stable in aqueous solutions over broad pH range; v) is chemically
stable when incubated with hepatocyte extracts, vi) is chemically
stable when incubated with hepatocyte cell lines; vii) shows 253
minutes plasma half-life when injected IP into mice; and viii)
shows no immediate toxicity over 24 hours when injected IP into
mice at 0.6 .mu.mole/per mouse and at 1.2 .mu.mole/per mouse and
also no toxicity when injected IP into mice at 0.3 .mu.mole/per
mouse twice daily for 21 days.
[0239] In other embodiments, a 15-PGDH inhibitor having formula
(IX) can include a compound with the following formula (IXa):
##STR00074## [0240] and pharmaceutically acceptable salts
thereof.
[0241] In still other embodiments, a 15-PGDH inhibitor having
formula (IX) can include a compound with the following formula
(IXb):
##STR00075## [0242] and pharmaceutically acceptable salts
thereof.
[0243] In other embodiments, the 15-PDHG inhibitor can comprise a
(+) or (-) optical isomer of a 15-PGDH inhibitor having formula
(IX). In still other embodiments, the 15-PDHG inhibitor can
comprise a mixture at least one of a (+) or (-) optical isomer of a
15-PGDH inhibitor having formula (IX). For example, the 15-PGDH
inhibitor can comprise a mixture of: less than about 50% by weight
of the (-) optical isomer of a 15-PGDH inhibitor having formula
(IX) and greater than about 50% by weight of the (+) optical isomer
of a 15-PGDH inhibitor having formula (IX), less than about 25% by
weight of the (-) optical isomer of a 15-PGDH inhibitor having
formula (IX) and greater than about 75% by weight of the (+)
optical isomer of a 15-PGDH inhibitor having formula (IX), less
than about 10% by weight of the (-) optical isomer of a 15-PGDH
inhibitor having formula (IX) and greater than about 90% by weight
of the (+) optical isomer of a 15-PGDH inhibitor having formula
(IX), less than about 1% by weight of the (-) optical isomer of a
15-PGDH inhibitor having formula (IX) and greater than about 99% by
weight of the (+) optical isomer of a 15-PGDH inhibitor having
formula (IX), greater than about 50% by weight of the (-) optical
isomer of a 15-PGDH inhibitor having formula (IX) and less than
about 50% by weight of the (+) optical isomer of a 15-PGDH
inhibitor having formula (IX), greater than about 75% by weight of
the (-) optical isomer of a 15-PGDH inhibitor having formula (IX)
and less than about 25% by weight of the (+) optical isomer of a
15-PGDH inhibitor having formula (IX), greater than about 90% by
weight of the (-) optical isomer of a 15-PGDH inhibitor having
formula (IX) and less than about 10% by weight of the (+) optical
isomer of a 15-PGDH inhibitor having formula (IX), or greater than
about 99% by weight of the (-) optical isomer of a 15-PGDH
inhibitor having formula (IX) and less than about 1% by weight of
the (+) optical isomer of a 15-PGDH inhibitor having formula
(IX).
[0244] In a still further embodiment, the 15-PDGH inhibitor can
consist essentially of or consist of the (+) optical isomer of a
15-PGDH inhibitor having formula (IX). In yet another embodiment,
the PDGH inhibitor can consist essentially of or consist of the (-)
optical isomer of a 15-PGDH inhibitor having formula (IX).
[0245] In other embodiments, a 15-PGDH inhibitor having formula
(VIII) can include a compound with the following formula (X):
##STR00076## [0246] and pharmaceutically acceptable salts
thereof.
[0247] Advantageously, the 15-PDGH inhibitor having formula (X) was
found to: i) inhibit recombinant 15-PGDH at 3 nM concentration; ii)
increase PGE.sub.2 production by cell lines at 20 nM; iii) is
chemically stable in aqueous solutions over broad pH range; iv) is
chemically stable when incubated with mouse, rat and human liver
extracts, v) shows 33 minutes plasma half-life when injected IP
into mice; viii) shows no immediate toxicity over 24 hours when
injected IP into mice at 50 mg/kg body weight, and ix) is soluble
in water (pH=3) at 1 mg/mL.
[0248] In other embodiments, a 15-PGDH inhibitor having formula (X)
can include a compound with the following formula (Xa):
##STR00077## [0249] and pharmaceutically acceptable salts
thereof.
[0250] In still other embodiments, a 15-PGDH inhibitor having
formula (X) can include a compound with the following formula
(Xb):
##STR00078## [0251] and pharmaceutically acceptable salts
thereof.
[0252] In other embodiments, the 15-PDHG inhibitor can comprise a
(+) or (-) optical isomer of a 15-PGDH inhibitor having formula
(X). In still other embodiments, the 15-PDHG inhibitor can comprise
a mixture at least one of a (+) or (-) optical isomer of a 15-PGDH
inhibitor having formula (X). For example, the 15-PGDH inhibitor
can comprise a mixture of: less than about 50% by weight of the (-)
optical isomer of a 15-PGDH inhibitor having formula (X) and
greater than about 50% by weight of the (+) optical isomer of a
15-PGDH inhibitor having formula (X), less than about 25% by weight
of the (-) optical isomer of a 15-PGDH inhibitor having formula (X)
and greater than about 75% by weight of the (+) optical isomer of a
15-PGDH inhibitor having formula (X), less than about 10% by weight
of the (-) optical isomer of a 15-PGDH inhibitor having formula (X)
and greater than about 90% by weight of the (+) optical isomer of a
15-PGDH inhibitor having formula (X), less than about 1% by weight
of the (-) optical isomer of a 15-PGDH inhibitor having formula (X)
and greater than about 99% by weight of the (+) optical isomer of a
15-PGDH inhibitor having formula (X), greater than about 50% by
weight of the (-) optical isomer of a 15-PGDH inhibitor having
formula (X) and less than about 50% by weight of the (+) optical
isomer of a 15-PGDH inhibitor having formula (X), greater than
about 75% by weight of the (-) optical isomer of a 15-PGDH
inhibitor having formula (X) and less than about 25% by weight of
the (+) optical isomer of a 15-PGDH inhibitor having formula (X),
greater than about 90% by weight of the (-) optical isomer of a
15-PGDH inhibitor having formula (X) and less than about 10% by
weight of the (+) optical isomer of a 15-PGDH inhibitor having
formula (X), or greater than about 99% by weight of the (-) optical
isomer of a 15-PGDH inhibitor having formula (X) and less than
about 1% by weight of the (+) optical isomer of a 15-PGDH inhibitor
having formula (X).
[0253] In a still further embodiment, the 15-PDGH inhibitor can
consist essentially of or consist of the (+) optical isomer of a
15-PGDH inhibitor having formula (X). In yet another embodiment,
the PDGH inhibitor can consist essentially of or consist of the (-)
optical isomer of a 15-PGDH inhibitor having formula (X).
[0254] It will be appreciated that the other 15-PGDH inhibitors can
be used in the methods described herein. These other 15-PGDH
inhibitors can include known 15-PGDH inhibitors including, for
example, tetrazole compounds of formulas (I) and (II),
2-alkylideneaminooxyacetamidecompounds of formula (I), heterocyclic
compounds of formulas (VI) and (VII), and pyrazole compounds of
formula (III) described in U.S. Patent Application Publication No.
2006/0034786 and U.S. Pat. No. 7,705,041;
benzylidene-1,3-thiazolidine compounds of formula (I) described in
U.S. Patent Application Publication No. 2007/0071699;
phenylfurylmethylthiazolidine-2,4-dione and
phenylthienylmethylthiazolidine-2,4-dione compounds described in
U.S. Patent Application Publication No. 2007/0078175;
thiazolidenedione derivatives described in U.S. Patent Application
Publication No. 2011/0269954; phenylfuran, phenylthiophene, or
phenylpyrrazole compounds described in U.S. Pat. No. 7,294,641,
5-(3,5-disubstituted phenylazo)-2-hydroxybenzene-acetic acids and
salts and lactones described in U.S. Pat. No. 4,725,676, and azo
compounds described in U.S. Pat. No. 4,889,846.
[0255] The 15-PGDH inhibitors described herein can be provided in a
pharmaceutical composition or cosmetic composition depending on the
pathological or cosmetic condition or disorder being treated. A
pharmaceutical composition containing the 15-PGDH inhibitors
described herein as an active ingredient may be manufactured by
mixing the derivative with a pharmaceutically acceptable carrier(s)
or an excipient(s) or diluting the 15-PGDH inhibitors with a
diluent in accordance with conventional methods. The pharmaceutical
composition may further contain fillers, anti-cohesives,
lubricants, wetting agents, flavoring agents, emulsifying agents,
preservatives and the like. The pharmaceutical composition may be
formulated into a suitable formulation in accordance with the
methods known to those skilled in the art so that it can provide an
immediate, controlled or sustained release of the 15-PGDH
inhibitors after being administered into a mammal.
[0256] In some embodiments, the pharmaceutical composition may be
formulated into a parenteral or oral dosage form. The solid dosage
form for oral administration may be manufactured by adding
excipient, if necessary, together with binder, disintegrants,
lubricants, coloring agents, and/or flavoring agents, to the
15-PGDH inhibitors and shaping the resulting mixture into the form
of tablets, sugar-coated pills, granules, powder or capsules. The
additives that can be added in the composition may be ordinary ones
in the art. For example, examples of the excipient include lactose,
sucrose, sodium chloride, glucose, starch, calcium carbonate,
kaolin, microcrystalline cellulose, silicate and the like.
Exemplary binders include water, ethanol, propanol, sweet syrup,
sucrose solution, starch solution, gelatin solution,
carboxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl
starch, methylcellulose, ethylcellulose, shellac, calcium
phosphonate and polypyrrolidone. Examples of the disintegrant
include dry starch, sodium arginate, agar powder, sodium
bicarbonate, calcium carbonate, sodium lauryl sulfate, stearic
monoglyceride and lactose. Further, purified talc, stearates,
sodium borate, and polyethylene glycol may be used as a lubricant;
and sucrose, bitter orange peel, citric acid, tartaric acid, may be
used as a flavoring agent. In some embodiments, the pharmaceutical
composition can be made into aerosol formulations (e.g., they can
be nebulized) to be administered via inhalation.
[0257] The 15-PGDH inhibitors described herein may be combined with
flavoring agents, buffers, stabilizing agents, and the like and
incorporated into oral liquid dosage forms such as solutions,
syrups or elixirs in accordance with conventional methods. One
example of the buffers may be sodium citrate. Examples of the
stabilizing agents include tragacanth, acacia and gelatin.
[0258] In some embodiments, the 15-PGDH inhibitors described herein
may be incorporated into an injection dosage form, for example, for
a subcutaneous, intramuscular or intravenous route by adding
thereto pH adjusters, buffers, stabilizing agents, relaxants,
topical anesthetics. Examples of the pH adjusters and the buffers
include sodium citrate, sodium acetate and sodium phosphate.
Examples of the stabilizing agents include sodium pyrosulfite,
EDTA, thioglycolic acid and thiolactic acid. The topical
anesthetics may be procaine HCl, lidocaine HCl and the like. The
relaxants may be sodium chloride, glucose and the like.
[0259] In other embodiments, the 15-PGDH inhibitors described
herein may be incorporated into suppositories in accordance with
conventional methods by adding thereto pharmaceutically acceptable
carriers that are known in the art, for example, polyethylene
glycol, lanolin, cacao butter or fatty acid triglycerides, if
necessary, together with surfactants such as Tween.
[0260] The pharmaceutical composition may be formulated into
various dosage forms as discussed above and then administered
through various routes including an oral, inhalational,
transdermal, subcutaneous, intravenous or intramuscular route. The
dosage can be a pharmaceutically or therapeutically effective
amount.
[0261] Therapeutically effective dosage amounts of the 15-PGDH
inhibitor may be present in varying amounts in various embodiments.
For example, in some embodiments, a therapeutically effective
amount of the 15-PGDH inhibitor may be an amount ranging from about
10-1000 mg (e.g., about 20 mg-1,000 mg, 30 mg-1,000 mg, 40 mg-1,000
mg, 50 mg-1,000 mg, 60 mg-1,000 mg, 70 mg-1,000 mg, 80 mg-1,000 mg,
90 mg-1,000 mg, about 10-900 mg, 10-800 mg, 10-700 mg, 10-600 mg,
10-500 mg, 100-1000 mg, 100-900 mg, 100-800 mg, 100-700 mg, 100-600
mg, 100-500 mg, 100-400 mg, 100-300 mg, 200-1000 mg, 200-900 mg,
200-800 mg, 200-700 mg, 200-600 mg, 200-500 mg, 200-400 mg,
300-1000 mg, 300-900 mg, 300-800 mg, 300-700 mg, 300-600 mg,
300-500 mg, 400 mg-1,000 mg, 500 mg-1,000 mg, 100 mg-900 mg, 200
mg-800 mg, 300 mg-700 mg, 400 mg-700 mg, and 500 mg-600 mg). In
some embodiments, the 15-PGDH inhibitor is present in an amount of
or greater than about 10 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg,
300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700
mg, 750 mg, 800 mg. In some embodiments, the 15-PGDH inhibitor is
present in an amount of or less than about 1000 mg, 950 mg, 900 mg,
850 mg, 800 mg, 750 mg, 700 mg, 650 mg, 600 mg, 550 mg, 500 mg, 450
mg, 400 mg, 350 mg, 300 mg, 250 mg, 200 mg, 150 mg, or 100 mg.
[0262] In other embodiments, a therapeutically effective dosage
amount may be, for example, about 0.001 mg/kg weight to 500 mg/kg
weight, e.g., from about 0.001 mg/kg weight to 400 mg/kg weight,
from about 0.001 mg/kg weight to 300 mg/kg weight, from about 0.001
mg/kg weight to 200 mg/kg weight, from about 0.001 mg/kg weight to
100 mg/kg weight, from about 0.001 mg/kg weight to 90 mg/kg weight,
from about 0.001 mg/kg weight to 80 mg/kg weight, from about 0.001
mg/kg weight to 70 mg/kg weight, from about 0.001 mg/kg weight to
60 mg/kg weight, from about 0.001 mg/kg weight to 50 mg/kg weight,
from about 0.001 mg/kg weight to 40 mg/kg weight, from about 0.001
mg/kg weight to 30 mg/kg weight, from about 0.001 mg/kg weight to
25 mg/kg weight, from about 0.001 mg/kg weight to 20 mg/kg weight,
from about 0.001 mg/kg weight to 15 mg/kg weight, from about 0.001
mg/kg weight to 10 mg/kg weight.
[0263] In still other embodiments, a therapeutically effective
dosage amount may be, for example, about 0.0001 mg/kg weight to 0.1
mg/kg weight, e.g. from about 0.0001 mg/kg weight to 0.09 mg/kg
weight, from about 0.0001 mg/kg weight to 0.08 mg/kg weight, from
about 0.0001 mg/kg weight to 0.07 mg/kg weight, from about 0.0001
mg/kg weight to 0.06 mg/kg weight, from about 0.0001 mg/kg weight
to 0.05 mg/kg weight, from about 0.0001 mg/kg weight to about 0.04
mg/kg weight, from about 0.0001 mg/kg weight to 0.03 mg/kg weight,
from about 0.0001 mg/kg weight to 0.02 mg/kg weight, from about
0.0001 mg/kg weight to 0.019 mg/kg weight, from about 0.0001 mg/kg
weight to 0.018 mg/kg weight, from about 0.0001 mg/kg weight to
0.017 mg/kg weight, from about 0.0001 mg/kg weight to 0.016 mg/kg
weight, from about 0.0001 mg/kg weight to 0.015 mg/kg weight, from
about 0.0001 mg/kg weight to 0.014 mg/kg weight, from about 0.0001
mg/kg weight to 0.013 mg/kg weight, from about 0.0001 mg/kg weight
to 0.012 mg/kg weight, from about 0.0001 mg/kg weight to 0.011
mg/kg weight, from about 0.0001 mg/kg weight to 0.01 mg/kg weight,
from about 0.0001 mg/kg weight to 0.009 mg/kg weight, from about
0.0001 mg/kg weight to 0.008 mg/kg weight, from about 0.0001 mg/kg
weight to 0.007 mg/kg weight, from about 0.0001 mg/kg weight to
0.006 mg/kg weight, from about 0.0001 mg/kg weight to 0.005 mg/kg
weight, from about 0.0001 mg/kg weight to 0.004 mg/kg weight, from
about 0.0001 mg/kg weight to 0.003 mg/kg weight, from about 0.0001
mg/kg weight to 0.002 mg/kg weight. In some embodiments, the
therapeutically effective dose may be 0.0001 mg/kg weight, 0.0002
mg/kg weight, 0.0003 mg/kg weight, 0.0004 mg/kg weight, 0.0005
mg/kg weight, 0.0006 mg/kg weight, 0.0007 mg/kg weight, 0.0008
mg/kg weight, 0.0009 mg/kg weight, 0.001 mg/kg weight, 0.002 mg/kg
weight, 0.003 mg/kg weight, 0.004 mg/kg weight, 0.005 mg/kg weight,
0.006 mg/kg weight, 0.007 mg/kg weight, 0.008 mg/kg weight, 0.009
mg/kg weight, 0.01 mg/kg weight, 0.02 mg/kg weight, 0.03 mg/kg
weight, 0.04 mg/kg weight, 0.05 mg/kg weight, 0.06 mg/kg weight,
0.07 mg/kg weight, 0.08 mg/kg weight, 0.09 mg/kg weight, or 0.1
mg/kg weight. The effective dose for a particular individual can be
varied (e.g., increased or decreased) over time, depending on the
needs of the individual.
[0264] In some embodiments, a therapeutically effective dosage may
be a dosage of 10 .mu.g/kg/day, 50 .mu.g/kg/day, 100 .mu.g/kg/day,
250 .mu.g/kg/day, 500 .mu.g/kg/day, 1000 .mu.g/kg/day or more. In
various embodiments, the amount of the 15-PGDH inhibitor or
pharmaceutical salt thereof is sufficient to provide a dosage to a
patient of between 0.01 .mu.g/kg and 10 .mu.g/kg; 0.1 .mu.g/kg and
5 .mu.g/kg; 0.1 .mu.g/kg and 1000 .mu.g/kg; 0.1 .mu.g/kg and 900
.mu.g/kg; 0.1 .mu.g/kg and 900 .mu.g/kg; 0.1 .mu.g/kg and 800
.mu.g/kg; 0.1 .mu.g/kg and 700 .mu.g/kg; 0.1 .mu.g/kg and 600
.mu.g/kg; 0.1 .mu.g/kg and 500 .mu.g/kg; or 0.1 .mu.g/kg and 400
.mu.g/kg.
[0265] Particular doses or amounts to be administered in accordance
with the present invention may vary, for example, depending on the
nature and/or extent of the desired outcome, on particulars of
route and/or timing of administration, and/or on one or more
characteristics (e.g., weight, age, personal history, genetic
characteristic, lifestyle parameter, severity of cardiac defect
and/or level of risk of cardiac defect, etc., or combinations
thereof). Such doses or amounts can be determined by those of
ordinary skill. In some embodiments, an appropriate dose or amount
is determined in accordance with standard clinical techniques. For
example, in some embodiments, an appropriate dose or amount is a
dose or amount sufficient to reduce a disease severity index score
by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,
19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95,
100% or more. For example, in some embodiments, an appropriate dose
or amount is a dose or amount sufficient to reduce a disease
severity index score by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70,
75, 80, 85, 90, 95, 100%. Alternatively or additionally, in some
embodiments, an appropriate dose or amount is determined through
use of one or more in vitro or in vivo assays to help identify
desirable or optimal dosage ranges or amounts to be
administered.
[0266] Various embodiments may include differing dosing regimen. In
some embodiments, the 15-PGDH inhibitor can be administered via
continuous infusion. In some embodiments, the continuous infusion
is intravenous. In other embodiments, the continuous infusion is
subcutaneous. Alternatively or additionally, in some embodiments,
the 15-PGDH inhibitor can be administered bimonthly, monthly, twice
monthly, triweekly, biweekly, weekly, twice weekly, thrice weekly,
daily, twice daily, or on another clinically desirable dosing
schedule. The dosing regimen for a single subject need not be at a
fixed interval, but can be varied over time, depending on the needs
of the subject.
[0267] For topical application, the composition can be administered
in the form of aqueous, alcoholic, aqueous-alcoholic or oily
solutions or suspensions, or of a dispersion of the lotion or serum
type, of emulsions that have a liquid or semi-liquid consistency or
are pasty, obtained by dispersion of a fatty phase in an aqueous
phase (O/W) or vice versa (W/O) or multiple emulsions, of a free or
compacted powder to be used as it is or to be incorporated into a
physiologically acceptable medium, or else of microcapsules or
microparticles, or of vesicular dispersions of ionic and/or
nonionic type. It may thus be in the form of a salve, a tincture,
milks, a cream, an ointment, a powder, a patch, an impregnated pad,
a solution, an emulsion or a vesicular dispersion, a lotion,
aqueous or anhydrous gels, a spray, a suspension, a shampoo, an
aerosol or a foam. It may be anhydrous or aqueous. It may also
comprise solid preparations constituting soaps or cleansing
cakes.
[0268] Pharmaceutical compositions including the 15-PGDH inhibitor
described herein can additionally contain, for example, at least
one compound chosen from prostaglandins, in particular
prostaglandin PGE.sub.1, PGE.sub.2, their salts, their esters,
their analogues and their derivatives, in particular those
described in WO 98/33497, WO 95/11003, JP 97-100091, JP 96-134242,
in particular agonists of the prostaglandin receptors. It may in
particular contain at least one compound such as the agonists (in
acid form or in the form of a precursor, in particular in ester
form) of the prostaglandin F.sub.2.alpha. receptor, such as for
example latanoprost, fluprostenol, cloprostenol, bimatoprost,
unoprostone, the agonists (and their precursors, in particular the
esters such as travoprost) of the prostaglandin E.sub.2 receptors
such as 17-phenyl PGE.sub.2, viprostol, butaprost, misoprostol,
sulprostone, 16,16-dimethyl PGE.sub.2, 11-deoxy PGE.sub.1, 1-deoxy
PGE.sub.1, the agonists and their precursors, in particular esters,
of the prostacycline (IP) receptor such as cicaprost, iloprost,
isocarbacycline, beraprost, eprostenol, treprostinil, the agonists
and their precursors, in particular the esters, of the
prostaglandin D.sub.2 receptor such as BW245C
((4S)-(3-[(3R,S)-3-cyclohexyl-3-isopropyl]-2,5-dioxo)-4-imidazolidinehept-
-anoic acid), BW246C
((4R)-(3-[(3R,S)-3-cyclohexyl-3-isopropyl]-2,5-dioxo)-4-imidazolidinehept-
-anoic acid), the agonists and their precursors, in particular the
esters, of the receptor for the thromboxanes A2 (TP) such as I-BOP
([1S-[1a,2a(Z),
3b(1E,3S),4a]]-7-[3-[3-hydroxy-4-[4-(iodophenoxy)-1-butenyl]-7-oxabicyclo-
-[2.2.1]hept-2-yl]-5-heptenoic acid).
[0269] Advantageously, the composition can include at least one
15-PGDH inhibitor as defined above and at least one prostaglandin
or one prostaglandin derivative such as for example the
prostaglandins of series 2 including in particular PGF.sub.2, and
PGE.sub.2 in saline form or in the form of precursors, in
particular of the esters (example isopropyl esters), their
derivatives such as 16,16-dimethyl PGE.sub.2, 17-phenyl PGE.sub.2
and 16,16-dimethyl PGF.sub.2, 17-phenyl PGF.sub.2a, prostaglandins
of series 1 such as 11-deoxyprostaglandin E1, 1-deoxyprostaglandin
E1 in saline or ester form, is their analogues, in particular
latanoprost, travoprost, fluprostenol, unoprostone, bimatoprost,
cloprostenol, viprostol, butaprost, misoprostol, their salts or
their esters.
[0270] The invention is further illustrated by the following
examples, which is not intended to limit the scope of the
claims.
Example 1
Analysis of Effect of SW033291 on Bone Marrow Function
[0271] This Example shows effects of SW033291 on bone marrow
function.
[0272] FIGS. 1(A-C) show analysis of bone marrow of wild-type mice
versus mice that are homozygous genetic knockouts for 15-PGDH
(PGDH-/- mice). Total bone marrow cellularity and percent of
Scal+/c-Kit+ cells in lineage negative (SKL) cells are the same in
both sets of mice. However, bone marrow from 15-PGDH-/- mice shows
an approximately 50% increase in numbers of hematopoietic colonies
generated when marrow is plated into methylcelluose. 15-PGDH
knockout mice are denoted by label PGDH mice and by label 15-PGDH.
WT denotes wild-type mice.
[0273] FIGS. 2(A-F) shows assays in which bone marrow is harvested
from a wild-type mouse, and incubated ex vivo on ice for 2 hours
with either SW033291 (0.5 .mu.M), or 1 .mu.M PGE2 or 1 .mu.M
16,16-dimethyl PGE2 (dmPGE2). Treated marrow is again then plated
into methylcellulose for counting of hematopoietic colonies.
SW033291 treated marrow again shows an approximately 50% increase
in the number of bone marrow derived colonies generated. Under
these conditions, a lesser increase is seen in marrow treated with
PGE2, and a slightly greater increase is seen in marrow treated
with dmPGE2.
[0274] FIGS. 3(A-C) show a study of C57BL/6J mice treated with IP
SW033291 administered in a vehicle of 10% Ethanol, 5% Cremophor EL,
85% D5W at a dose of 5 mg/kg or 20 mg/kg. Panel A shows mouse bone
marrow cellularity, white blood count (wbc), red blood count (rbc)
and platelets counts. Panel B shows percent of Scal+/c-Kit+ cells
in lineage negative (SKL) cells are unchanged in SW033291 treated
mice. Panel C shows that marrow from SW033291 treated mice gives
rise to approximately 30% increase in numbers of hematopoietic
colonies generated when marrow is plated into methylcelluose.
Experimental conditions are noted on the figure.
[0275] FIGS. 4(A-B) show analysis of marrow from CD45.2 antigen
marked C57BL/6J mice that were treated with SW033291 5 mg/kg IP
daily for 3 doses in a vehicle of 10% Ethanol, 5% Cremophor EL, 85%
D5W or that were treated with vehicle alone. On day 3 mice were
sacrificed, marrow flushed and mixed at a 1:1 ratio with vehicle
treated CD45.1 marrow. 2 million whole BM cells were injected into
the tail vein of lethally irradiated CD45.1 mice and percent
chimerism measured via flow cytometry at weeks 8, 12, 16. As shown,
at weeks 12 and 16 the percent blood chimerism of CD45.2 marked
cells was significantly increased in recipient mice whose CD45.2
marked marrow was harvested from SW033291 treated donor mice, as
opposed to vehicle control treated donor mice. In other words,
marrow from SW033291 treated mice demonstrated long term increased
fitness in competition with control marrow. In particular, at week
16 CD45.2 harvested from SW033291 treated mice show a significant
increase in contribution to B and T cell populations, suggesting
marrow from SW033291 treated mice promotes earlier reconstitution
of lymphoid populations and earlier return to immune
competence.
[0276] In an additional study, C57BL/6J mice are irradiated with
11Gy on day 0, followed by treatment with SW033291 5 mg/kg IP twice
daily (bid) in a vehicle of 10% Ethanol, 5% Cremophor EL, 85% D5W,
or with vehicle only for 21 days. Mice treated with vehicle or with
SW033291 all receive an allograft of marrow from a donor C57BL/6J
mouse at a dose of either 100,000 cells, 200,000 cells, 500,000
cells. 3 control and 3 SW033291 mice are assessed under each
condition. The experimental design is depicted in FIG. 50.
[0277] Table 1 shows the number of surviving mice in each cohort
over the first 19 days of study. Under the conditions of the mouse
colony during this study, control mice receiving 100,000-500,000
cells are all dead between days 4-13 of study. In contrast, two
SW033291 treated mice receiving 500,000 cells remain alive on day
19 of the study and are presumed to have full hematopoietic
reconstitution. Thus treatment with the 15-PGDH inhibitor SW033291
promoted survival of mice receiving a bone marrow transplant, an
observation consistent with SW033291 enabling more rapid and
complete hematopoietic reconstitution in the transplanted mice.
Other 15-PGDH inhibitors with activity similar to SW033291 would be
predicted to have similar activity in supporting hematopoietic
reconstitution. Treatment with SW033291 also enabled mice to be
successfully transplanted with a smaller inoculum of donor bone
marrow than the 1,000,000 cells that are standardly needed. These
observations suggest SW033291, as well as other similar 15-PGDH
inhibitors, is able to support successful transplantation with
smaller numbers of donor stem cells. Such activity would be of
particular utility in settings, such as transplantation with
umbilical cord stem cells, in which donor cell numbers are limited.
Improved survival of transplanted mice treated with SW033291
suggests efficacy of SW033291, and of similar 15-PGDH inhibitors,
as replacements for, or in enabling decreased use of, other
treatments or growth factors commonly employed in support of
patients receiving bone marrow, hematopoietic stem cell, and cord
blood stem cell transplants. Improved survival of transplanted mice
treated with SW033291 is consistent with SW033291, and by extension
other similar 15-PGDH inhibitors, having activity in reducing
infections in the transplanted mice, and/or in promoting recovery
of mice intestines from damage by radiation, and/or in reducing
pulmonary toxicity from radiation.
TABLE-US-00001 TABLE 1 Mouse 13- 14- 15- 16- 17- 18- 19- 20- 21-
22- 23- 24- 25- 26- 1- survival Mar Mar Mar Mar Mar Mar Mar Mar Mar
Mar Mar Mar Mar Mar Apr Cell Day Day Day Day Day Day Day Day Day
Day Day Day Day Day Day Treatment number 0 1 2 3 4 5 6 7 8 9 10 11
12 13 ... 19 Control 1 .times. 10{circumflex over ( )}5 3 3 3 2 0
Control 2 .times. 10{circumflex over ( )}5 3 3 3 3 3 2 1 0 Control
5 .times. 10{circumflex over ( )}5 3 3 3 3 3 3 2 1 1 1 1 1 1 0
SW033291 1 .times. 10{circumflex over ( )}5 3 3 3 3 2 1 0 SW033291
2 .times. 10{circumflex over ( )}5 3 3 3 3 3 3 2 2 2 2 2 2 1 0
SW033291 5 .times. 10{circumflex over ( )}5 3 3 3 3 3 3 3 3 2 2 2 2
2 2 ... 2
Example 2
Analysis of Effect of SW033291 on Radiation Survival
[0278] This Example shows studies of the effect of SW033291 in mice
receiving whole body irradiation.
[0279] Table 2 shows the results of a study of 15 week old C57BL/6J
female mice irradiated with 7Gy, 9Gy, or 11Gy, and receiving daily
SW033291 5 mg/kg IP in a vehicle of 10% Ethanol, 5% Cremophor EL,
85% D5W for 7 doses, or receiving vehicle alone. The table shows
the number of mice surviving on sequential days of the study. Under
the conditions of the mouse colony during this experiment, mice
receiving a lethal dose of 11Gy lived 48 hours longer if treated
with SW033291 than if receiving vehicle control, with control mice
all dead on day 8; whereas SW033219 treated mice were all dead on
day 10.
TABLE-US-00002 TABLE 2 10/2 10/3 10/4 10/5 10/6 10/7 10/8 10/9
10/10 10/11 10/12 10/13 10/23 Radiation Treatment Day Day Day Day
Day Day Day Day Day Day Day Day Day Day Dose Arm 0 5 6 7 8 9 10 11
12 13 14 15 16 25 7 Gy Saline 3 3 3 3 3 3 3 3 3 3 3 3 Looks Healthy
SW033291 3 3 3 3 3 3 3 3 3 3 3 3 Looks Healthy 9 Gy Saline 3 3 3 3
3 3 3 3 2 2 1 0 SW033291 3 3 3 3 3 3 3 3 2 2 1 0 11 Gy Saline 3 3 3
2 0 SW033291 3 3 3 3 3 2 0
[0280] Table 3 shows the number of mice surviving on sequential
days of a study of mice treated at 11Gy treated with either vehicle
control or with SW033291 IP, in a vehicle of 10% Ethanol, 5%
Cremophor EL, 85% D5W, with SW033291 administered either at 5 mg/kg
daily for 7 days, 5 mg/kg daily throughout the study, or at 5 mg/kg
twice daily for 7 days. Again mice treated with SW033291 on any of
these dosing schedules live on average 1-2 days longer than mice
receiving vehicle control. The activity of SW033291 in promoting
resistance to toxic effects of radiation may extend to SW033291 and
other similar 15-PGDH inhibitors in promoting resistance to other
similar toxic insults including but not limited to Cytoxan,
fludarabine, chemotherapy and immunosuppressive therapy.
TABLE-US-00003 TABLE 3 Friday Wed. Thurs. Friday Saturday Sunday
Monday Treatment 12-Oct 17-Oct 18-Oct 19-Oct 20-Oct 21-Oct 22-Oct
Condition Day 0 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10 11 Saline (7 2
2 2 2 0 Gy days, 1 dose daily) 11 SW033291 3 3 3 3 2 1 0 Gy (1
dose/daily) for 7 days 11 SW033291 3 3 3 3 3 2 0 Gy (1 dose/daily,
continous every day) 11 Saline (7 3 3 3 2 1 0 0 Gy days, 2
dose/daily) 11 SW033291 (7 3 3 3 3 3 2 0 Gy days, 2 dose daily)
Example 3
Analysis of Effect of SW033291 on Survival of Mice Following Bone
Marrow Transplant
[0281] FIG. 5 shows enhanced survival in mice receiving a bone
marrow transplant and also administered the 15-PGDH inhibitor
SW033291. FIG. 5A shows the design of the study. Mice were
irradiated with a bone marrow ablative dose of 11Gy on day 0,
followed by administration of SW033291 in 5 mg/kg twice daily by
intraperitoneal injection in a vehicle of 10% Ethanol, 5% Cremophor
EL, 85% D5W at a concentration of 125 .mu.g/200 .mu.l. A matched
control cohort received injections with vehicle only. On day one
mice received an infusion of donor marrow at doses of 100,000
cells; 200,000 cells; or 500,000 cells. FIG. 5B shows graphical
survival curves for mice transplanted with 100,000 donor cells.
FIG. 5C shows graphical survival curves for mice transplanted with
200,000 donor cells. FIG. 5D shows graphical survival curves for
mice transplanted with 500,000 donor cells. Table 4 shows tabular
survival data for all mice in the study on study day 30. Among mice
receiving 100,000 donor cells, all mice succumb, but SW033291
treated mice show an approximate doubling of median survival. Among
mice receiving 200,000 donor cells, all control mice were dead by
day 12. In contrast, all mice receiving 200,000 donor cells plus
SW033291 survived at 30 days of observation and were successfully
engrafted. Among mice receiving 500,000 donor cells, control mice
showed a 37.5% mortality; whereas, mice receiving SW033291 again
all survived.
TABLE-US-00004 TABLE 4 Dose Vehicle Survival SW033291 Survival
1*10.sup.5 0/8 1/8 2*10.sup.5 0/8 8/8 5*10.sup.5 5/8 8/8
[0282] FIG. 6 shows a set of studies conducted on lethally
irradiated mice that received 500,000 donor marrow cells and were
treated with either SW033291 at 5 mg/kg intraperitoneal dose twice
daily or with vehicle control, in a design otherwise identical to
that used for the studies of FIG. 5. FIG. 6A shows measurements on
blood and bone marrow on day 5 after transplant, with FIG. 6B
showing that SW033291 treated mice have significantly higher total
white count and FIG. 6C showing that SW033291 treated mice have
significantly higher total platelet count. The star symbol denotes
P<0.05.
[0283] FIG. 7A shows measurements on blood and bone marrow on day 8
after transplant. FIG. 7B shows that SW033291 treated mice have
significantly higher platelet count than control, with drug treated
mice having 77,000 platelets compared to control mice having 39,500
platelets. The star symbol denotes P<0.05.
[0284] FIG. 8A shows measurements on blood and bone marrow on day
12 after transplant. FIG. 8B shows that SW033291 treated mice have
significantly higher neutrophil counts, with drug treated mice
having 332 neutrophils compared to control mice having 125
neutrophils. FIG. 8C shows that on day 12 after transplant,
SW033291 treated mice have significantly higher hemoglobin count
than controls, with drug treated mice having hemoglobin level of
11.58 and control mice having hemoglobin level of 8.3.
Additionally, FIG. 8D shows SW033291 treated mice also have
significantly greater total white counts than control mice. The
star symbol denotes P<0.05.
[0285] FIG. 9A shows measurements on blood and bone marrow on day
18 after transplant. SW033291 treated mice have significantly
higher total white count (FIG. 9B), lymphocyte count (FIG. 9C), and
neutrophil count (FIG. 9D), with drug treated mice having 835
neutrophils and control mice having 365 neutrophils (FIG. 9D). In
comparison with counts on day 12, administration of SW033291
accelerates recovery of neutrophil counts by nearly 6 days (FIG. 8B
versus FIG. 9D). FIG. 8E also shows that on day 18 drug treated
mice have significantly higher platelet counts than control mice.
Last, day 18, drug treated mice have nearly 4-fold increased
percentage (FIG. 8F) and total numbers (FIG. 9G) of SKL marked bone
marrow stem cells than do control mice, with drug treated mice
having a mean of 4127 SKL marked bone marrow cells compared to
control mice having a mean of 967 SKL marked bone marrow cells. The
star symbol denotes P<0.05.
[0286] FIG. 10(A) shows measurement of PGE2 (pg of PGE2/mg tissue
protein) in 4 different mouse tissues (colon, bone marrow, liver,
lung) across time following IP injection of SW033291 at 10 mg/kg.
Blue bar represents baseline at time 0, and red bars represent time
course of PGE2 concentration from 1-12 hours following SW033291
injection. FIG. 10(B) shows time course of PGE2 in control mice
injected with vehicle only.
[0287] FIG. 11 shows a schema of an experiment in which mice are
lethally irradiated (IR) and 12 hours later receive a transplant
(BMT) with CFSE dye labeled bone marrow cells (BM), and the numbers
of transplanted cells that home and survive in the bone marrow of
the recipient mice are then determined by FACS at 16 hours
post-transplant. In different arms of the experiment mice are
treated with vehicle, with SW033291 (10 mg/kg IP), or with SW033291
(10 mg/kg IP) plus Indomethacin. Drugs are administered following
radiation, following the transplant, and again at 8 hours after the
transplant.
[0288] FIG. 12 shows a graph illustrating the percent of CFSE dye
labeled cells that have homed to the bone marrow of mice treated as
per the schema described in FIG. 11. Treating mice with SW033291
concurrent with and following the bone marrow transplant increases
numbers of homed cells in the recipient mouse bone marrow 3-fold.
The figure further shows that the effect of SW033291 is near
completely blocked by indomethacin (Indo+SW0), an inhibitor of COX
enzymes the produce prostaglandins. This is consistent with the
effect of SW033291 being mediated through the inhibition of 15-PGDH
and through the resulting increase in tissue prostaglandins.
[0289] FIG. 13 shows a schema of an experiment in which mice are
lethally irradiated (IR) and 12 hours later receive a transplant
(BMT) with CFSE dye labeled bone marrow cells (BM), and numbers of
transplanted cells that home and survive in the bone marrow of the
recipient mice are then determined by FACS at 16 hours
post-transplant. In different arms of the experiment mice are
treated with vehicle, with SW033291 (10 mg/kg IP), with SW033291
(10 mg/kg IP) plus an antagonist of PGE2 receptor EP2
(PF-04418948), or with SW033291 (10 mg/kg IP) plus an antagonist of
PGE2 receptor EP4 (L-161,982). Drugs are administered following
radiation, following the transplant, and again at 8 hours after the
transplant.
[0290] FIG. 14 shows a graph illustrating the percent of CFSE dye
labeled cells that have homed to the bone marrow of mice treated as
per the schema described in FIG. 13. Treating mice with SW033291
concurrent with and following the bone marrow transplant increases
numbers of homed cells in the recipient mouse bone marrow 2-fold.
The figure further shows that the effect of SW033291 is near
completely blocked by an antagonist to the EP4 receptor (EP4+SW0),
and is partially blocked by an antagonist of the EP2 receptor
(EP2+SW0). This is consistent with the effect of SW033291 being
mediated through the inhibition of 15-PGDH and through the
resulting increase of tissue prostaglandins, including PGE2.
[0291] In another experiment, mice were injected with SW033291
twice daily IP at 10 mg/kg for 5 doses. 2 hours following the last
dose bone marrow is harvested and sorted into SKL marked cells that
are hematopoietic stem cell enriched and into CD45 (-) cells that
are bone marrow stroma cells. RNA was extracted and gene expression
determined relative to levels in mice injected with vehicle
control.
[0292] FIGS. 15(A-B) show graph illustrating induction of gene
expression in bone marrow SKL cells and bone marrow stromal cells
of SW033291 treated mice. Mice administered SW033291 show a 3 fold
induction in RNA expression of CXCL12 and SCF in bone marrow SKL
cells, and show a greater than 4-fold induction of CXCL12 and SCF
in CD45(-) bone marrow stromal cells.
[0293] FIG. 16 illustrates a schema of an experiment in which
immune deficient NSG mice are lethally irradiated (IR) and 12 hours
later receive a transplant with CFSE dye labeled buffy coat cells
from human umbilical cord blood (UCB), and number of transplanted
cells that home and survive in the bone marrow of the recipient
mice are then determined by FACS at 16 hours post-transplant. In
different arms of the experiment mice are treated with vehicle or
with SW033291. Drugs are administered following radiation,
following the transplant, and again at 8 hours after the
transplant.
[0294] FIG. 17 illustrates a graph showing the percent of CFSE dye
labeled human umbilical cord buffy coat cells that have homed to
the bone marrow of mice treated as per the schema described in FIG.
18. Treating mice with SW033291 at the time of and following the
transplant with buffy coat from human umbilical cord blood (UCB)
increases numbers of homed human cells in the recipient mouse bone
marrow nearly 2-fold.
Example 3
[0295] FIG. 18 shows the design of a study in which 4 groups of
mice (4 mice per group) were administered either;
[0296] a) vehicle control,
[0297] b) 2.5 mg/kg of 15-PGDH inhibitor (+) SW209415 twice daily
IP for 7 doses,
[0298] c) recombinant human G-CSF 250 ug/kg subcutaneously once
daily for 4 doses
[0299] d) the combination of 2.5 mg/kg of 15-PGDH inhibitor (+)
SW209415 twice daily IP for 7 doses plus recombinant human G-CSF
250 ug/kg subcutaneously once daily for 4 doses (with the daily
dose of G-CSF being administered coincident with a dose of (+)
SW209415) 3 hours following the last drug dose mice were sacrificed
for analysis.
[0300] FIG. 19 shows results in each arm for measurement of total
white cells, neutrophils, and lymphocytes.
[0301] Neutrophil counts approximately double following
administration of G-CSF, approximately doubled following
administration of (+) SW209415, and increased 3 to 4-fold after
administration of the combination of G-CSF plus (+) SW209415.
[0302] FIG. 20 shows results in each arm for measurement of red
cells, hematocrit, hemoglobin, and platelets.
[0303] FIG. 21 shows results in each arm for measurement in
peripheral blood of circulating SKL (Sca-1+; C-kit+; Lin-) marked
cells that are enriched for hematopoietic stem cells. The results
show first that in this study G-CSF increased the frequency of
peripheral blood SKL cells (in Lin-cells) by 33%. (+) SW209415 when
administered alone showed no effect on frequency of peripheral
blood stem cells. However, the combination of G-CSF plus (+)
SW209415 increased by 2 to 3-fold the frequency of peripheral blood
SKL cells (in Lin-cells). This result understates the absolute
magnitude of this effect, as the combination of G-SCF plus (+)
SW209415 also increases the total white count, and hence the
absolute number of the Lin-population. Thus, G-CSF increased the
number of peripheral blood SKL cells by approximately 50%. (+)
SW209415 when administered alone showed no effect on number of
peripheral blood stem cells. However, the combination of G-CSF plus
(+) SW209415 increased the number of peripheral blood SKL cells by
approximately 3.5-fold, showing a marked increased effect of the
combination.
[0304] FIG. 22 shows results in each arm for measurement of SKL
cells in mice bone marrow. The show that bone marrow SKL marked
cells are increased by (+) SW209415, further increased by G-CSF,
and further increased by the combination of (+) SW209415 plus
G-CSF.
Example 4
[0305] In this Example, recipient mice lethally irradiated (11 Gy)
followed by BMT (500 k total cells) 16 hours post IR were
administered either:
[0306] a) Vehicle treated (2.times. daily).
[0307] b) 250 .mu.g/kg GCSF (1.times. daily, subcutaneous
administration, starting 24 hours post BMT)
[0308] c) 2.5 mg/kg SW209415 (+) (2.times. daily, 18 days, starting
post IR).
[0309] d) 250 .mu.g/kg hGCSF (1.times. daily, 17 days)+2.5 mg/kg
SW209415(+) (2.times. daily, 18 days).
[0310] Mice were bled via submandibular vein at days 8, 12, and 18
post transplant for CBC using the Hemavet 850 fs and sacrificed at
day 18 to assess marrow cellularity and SKL frequency. Blood counts
were tabulated graphically as illustrated in FIGS. 24-26 with error
bars corresponding to standard error of the means and compared
using 2-tailed t-tests.
SW209415 is Synergistic with GCSF in Promoting Hematopoietic
Recovery after BMT
[0311] Granulocyte colony stimulating factor (G-CSF, clinical names
Lenograstim and Filgrastim) is an FDA approved glycoprotein used
both pre-transplantation for stem cell priming purposes, and post
transplantation to enhance stem cell engraftment and minimize
mortality associated with prolonged neutropenia. Its use has been
shown to accelerate granulocyte engraftment by 1-6 days following
BMT compared to control. G-CSF occurs naturally and is responsible
for mounting the neutrophil activation in response to stress or
infection by binding to its receptor G-CSFR. Several tissues can
produce G-CSF upon stimulation by inflammatory mediators such as
LPS, TNF-.alpha., and IL-17, resulting in G-CSF release into the
bloodstream which stimulates neutrophil production within, and
mobilization from the bone marrow. In addition to neutrophil
mobilization, G-CSF has been implication in mobilizing HSCs from
marrow.
[0312] The use of SW209415 clinically in recipients of BMT would
need validation in clinical models in comparison and in combination
with GCSF. To determine SW209415's ability to promote hematopoietic
recovery following BMT independently as well as synergistically
with GCSF, we established the bone marrow transplant model
described above in which recipient mice receiving 500 k total bone
marrow cells were assessed for peripheral blood recovery at 3 time
points post transplant. FIGS. 23-26 show that compared to vehicle
treatment, SW209415, GCSF, and the combination treatment recovered
neutrophils significantly faster at Days 8, 12, and 18 post BMT,
but that the combination treatment demonstrated a synergistic
effect. Neutrophil counts at Day 8-150 (Veh), 339(415), 295(GCSF),
466-(4+G), Day 12-427(Veh) 839 (415), 982 (GCSF), 1390 (4+G), and
Day 18-649 (Veh), 1370 (415), 1518 (GCSF), 1703 (4+G) all
demonstrated an enhanced effect when the agents were used in
combination, showing the two compounds promote recovery via
independent mechanisms. Additionally we showed the frequency of SKL
cells (enriched for true HSCs) was higher in the combination
treated group compared to either therapeutic alone.
[0313] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
scope of the invention encompassed by the appended claims. All
patents, publications and references cited in the foregoing
specification are herein incorporated by reference in their
entirety.
* * * * *